{"articleList":{"__iterable":"Map","data":[["state",{"__iterable":"Map","data":[["isLoading",false]]}],["edges",null],["firstCursor",null],["lastCursor",null],["totalCount",null],["authFields",{"__iterable":"Map","data":[]}],["stats",{"__iterable":"Map","data":[["repliedCount",0],["notRepliedCount",0]]}]]},"replyList":{"__iterable":"Map","data":[["state",{"__iterable":"Map","data":[["isLoading",true]]}],["edges",null],["firstCursor",null],["lastCursor",null],["totalCount",null]]},"articleDetail":{"__iterable":"Map","data":[["state",{"__iterable":"Map","data":[["isLoading",false],["isAuthLoading",false],["isReplyLoading",false]]}],["data",{"__iterable":"Map","data":[["article",{"__iterable":"Map","data":[["replyCount",0],["references",{"__iterable":"List","data":[{"__iterable":"Map","data":[["type","LINE"]]}]}],["replyRequestCount",1],["user",{"__iterable":"Map","data":[["id","j4S8C_LwU7UmsX_4j0QjD-daIznoRLQESMzQqqF2opbLge8P8"],["name","果斷的長濱萊安"],["avatarUrl",null],["level",0]]}],["hyperlinks",{"__iterable":"List","data":[{"__iterable":"Map","data":[["title","Standard PLA+, Snow White, 1.75mm"],["url","https://sabzkadeh.com/3dfuel/?utm_content=119&utm_campaign=001"],["summary","Architectural and engineering models look fantastic in Snow White, one of our most popular colors of Standard PLA+ 3D printer filament."],["topImageUrl",null],["error",null]]}]}],["text","https://sabzkadeh.com/3dfuel/?utm_content=119&utm_campaign=001"],["replyRequests",{"__iterable":"List","data":[{"__iterable":"Map","data":[["id","3w1c8bkbi05b4__j4S8C_LwU7UmsX_4j0QjD-daIznoRLQESMzQqqF2opbLge8P8__RUMORS_LINE_BOT"],["reason",""],["positiveFeedbackCount",0],["negativeFeedbackCount",0]]}]}],["id","3w1c8bkbi05b4"],["createdAt","2023-01-22T02:57:22.715Z"]]}],["replyConnections",{"__iterable":"List","data":[]}],["relatedArticles",{"__iterable":"List","data":[{"__iterable":"Map","data":[["replyCount",1],["references",{"__iterable":"List","data":[{"__iterable":"Map","data":[["type","LINE"]]}]}],["replyRequestCount",1],["user",{"__iterable":"Map","data":[["id","j4S8C_9GNQGlQwAl1lpQmVJcTheQRtm0SiyV2ggNVwYUoi4PA"],["name","斯文的三重桃樂斯"],["avatarUrl",null],["level",0]]}],["hyperlinks",{"__iterable":"List","data":[{"__iterable":"Map","data":[["title","Flipboard on Flipboard"],["url","http://flip.it/TDnC4H"],["summary","@flipboard | Official Flipboard accountYour hand gestures can help make you more charismaticResearch into public speakers suggests hand gestures can powerfully change the way you are perceived. David Robson explains.

Next time you watch a TED talk or a political speech, take a moment to look closely at the speaker’s hand movements. Is the motion slow or energetic? Is it subtle or …CharismaJF Kennedy's childhood photos published – in picturesImages have been released as part of a year-long effort by the John F Kennedy Presidential Library and Museum to make 1,700 vulnerable nitrate negative photographs available online. They provide a glimpse of the Kennedy family from before JFK’s birth through to the early 1950sJohn F. KennedyHow Russia’s Skate Culture Found Its Own Unique Sense of StyleThough they were initially influenced by America, skaters from Moscow and beyond now present a truly local take on fashion.

Some people will tell you that the internet killed style subcultures—or a least turned them into a global, universally accessible phenomenon. We say that’s bullshit. Even as …Ralph LaurenWhere a Sore Throat Becomes a Death SentenceKIGALI, Rwanda — Neighbors whisper that she is pregnant, a disgrace for a young, unmarried woman. The rumors mortify her. She hates her swollen belly.

But Florence Ndimubakunzi is not pregnant. Her heart is failing. It pumps so poorly that blood backs up in her veins, bloating her liver and spleen, …Death PenaltyHow NAFTA Destroyed the Mexican DietThis story was originally published on Civil Eats.

When Alyshia Gálvez’s mother was growing up in California in the 1950s, she traveled to Michigan every winter to visit family. When she returned home, she would march into a grocery store, buy a head of lettuce, and eat it while sitting on the curb …Public HealthWhy a ‘Memory Town’ Is Coming to Your Local Strip MallWeeks after opening near San Diego, a model town for treating dementia is set to be replicated around the U.S.

On August 13, a brand-new town in Southern California welcomed its first residents. They trickled through the doors of a generic beige warehouse on a light-industrial stretch of Main Street …San DiegoSolving the genome puzzleWith advances in gene technology helping to diagnose very rare diseases, has the new era of personalised medicine finally arrived?

Evie Walker sits on her mother’s lap, playing a game she never grows tired of: turning her mother’s hand over and over, stroking and examining it. When she takes a break …The Re-reinvention of Chris(tine and the Queens)In her third act, France’s Héloïse Letissier is playfully muddying the gender lines. But under the name Chris, she continues to make pop’s brashest, queerest records with absolute clarity.

A petite Chris (née Christine and the Queens) lounges in New York’s Standard Hotel. Dressed in a white polo …Short HairThere are now more $100 bills than $1 bills in the worldA funny thing happened on the way to a world of cryptocurrencies and mobile payments. Cash became more popular than ever. The main reason? The one hundred dollar bill.

In 2017, for the first time ever, the one hundred dollar bill became the most popular US bill in circulation, beating out the one …Mobile PaymentsThe epic rise and fall of the name HeatherWhen people think of endangered species, they tend to think of the giant panda, the blue whale, or maybe the snow leopard. We need to add another group to the list: women named Heather.

A Quartz analysis of US baby-name data that goes back to 1880 finds that no name in history has become so popular …Giant PandasSilicon Valley thinks everyone feels the same six emotionsThe world is being flooded with technology designed to monitor our emotions. Amazon’s Alexa is one of many virtual assistants that detect tone and timbre of voice in order to better understand commands. CCTV cameras can track faces through public space, and supposedly detect criminals before they …Papua New GuineaIce Surveys and Neckties at Dinner: Here’s Life at an Arctic OutpostWant climate news in your inbox? Sign up here for Climate Fwd:, our email newsletter.

For the past 40 years, thanks to satellite measurements, scientists have known that sea ice coverage in the Arctic is shrinking. Global warming has reduced the extent of ice in the region in summer, when it is at its …The ArcticDaily Edition Top 10 (September 17, 2018)The controversy surrounding Supreme Court nominee Brett Kavanaugh and Dr. Christine Blasey Ford, Kavanaugh’s former classmate who has accused him of sexually assaulting her when they were teenagers, escalated this week. Congressional Republicans are reportedly offering to hold the hearing Wednesday …U.S. Supreme CourtHow Puberty Kills Girls’ ConfidenceIn their tween and teenage years, girls become dramatically less self-assured—a feeling that often lasts through adulthood.

The change can be baffling to many parents: Their young girls are masters of the universe, full of gutsy fire. But as puberty sets in, their confidence nose-dives, and those …PollingHow Far Have We Really Come Since Anita Hill?Christine Blasey Ford’s allegations against Brett Kavanaugh present a test that the Senate very well may not pass.

With a Supreme Court nominee’s confirmation hearings thrown into chaos by accusations of sexual misconduct, the Brett Kavanaugh drama feels like a familiar spectacle. During Clarence …Brett KavanaughThe Man Behind Trump’s ‘Invisible Wall’USCIS Director Lee Francis Cissna is the son of an immigrant, the son-in-law of a refugee and a man who says he’s just obsessed with the fair implementation of laws. So why is he making them so much harder for immigrants?

This summer, as anger over the separation of migrant families at the border …Homeland SecuritySee How the Fall Equinox Is Celebrated Around the WorldCulture & ExplorationPhoto Gallery

From early-morning gatherings at Stonehenge to torch-bearing revelers parading through a city in Spain, people around the world herald the seasonal change in myriad ways.

This summer, as anger over the separation of migrant families at the border …Homeland SecuritySee How the Fall Equinox Is Celebrated Around the WorldCulture & ExplorationPhoto Gallery

From early-morning gatherings at Stonehenge to torch-bearing revelers parading through a city in Spain, people around the world herald the seasonal change in myriad ways.

As the autumn equinox approaches, days begin to shorten in the Northern Hemisphere, and people …National Parks"],["topImageUrl","https://cdn.flipboard.com/uploads/avatar/7e211185df7384dc5032a1b22b634616c56c1238.jpeg"],["error",null]]}]}],["text","廢墟、童話城堡都有！彰化8個人氣「免費」好拍景點\n\nhttp://flip.it/dqxAaY"],["id","5410467194193-rumor"],["createdAt","2016-12-27T04:55:00.000Z"]]},{"__iterable":"Map","data":[["replyCount",1],["references",{"__iterable":"List","data":[{"__iterable":"Map","data":[["type","LINE"]]}]}],["replyRequestCount",2],["user",null],["hyperlinks",{"__iterable":"List","data":[{"__iterable":"Map","data":[["title","Edward M. Mautner Oral History"],["url","http://www.jsc.nasa.gov/history/oral_histories/NASA_HQ/Experts/MautnerEM/MautnerEM_3-22-12.htm"],["summary","NASA Johnson Space Center \n Oral History Project\n Edited Oral History Transcript\n Edward \n M. Mautner\n Interviewed by Jennifer Ross-Nazzal\n Chantilly, Virginia – 22 March 2012\n Ross-Nazzal: \n Today is March 22, 2012. This interview with Ed Mautner is being conducted \n at the [Smithsonian National Air and Space Museum] Steven Udvar-Hazy \n Center in Chantilly, Virginia, for the JSC Oral History Project. Interviewer \n is Jennifer Ross-Nazzal, assisted by Rebecca Wright. Thanks again \n for taking time out of your schedule.\n Mautner: \n You’re welcome, it’s my pleasure to do it.\n Ross-Nazzal: \n If you would, tell us about the state of [OV (orbiter vehicle)-101] \n Enterprise in 2003 before she was restored.\n Mautner: \n My familiarity with Enterprise was quite limited before I began work \n on it. When we received it, it had been stored both outdoors and in \n a non-climate-controlled hangar since the mid-80s, and a lot of deterioration \n of the surface coatings—the paint, the markings—had taken \n place. Woodpeckers had gotten up onto the vertical stabilizer and \n pecked a lot of holes into the polyurethane foam that they used as \n a mockup for tiles. I don’t recall the number of holes, but \n there were quite a few of them. \n And then just about every form of creature had made the Enterprise \n their habitat and playground. Birds had gotten into it; bugs had gotten \n into it. Hornets or wasps had made their little mud nests in the landing \n gear wells. They made it out of this beautiful red Virginia clay dirt \n that surrounds the area. In fact, when we started to clean them it \n almost seemed as if the aircraft was bleeding, the soil they made \n those nests out of was so red.\n It had basically suffered a lot of inattention. It hadn’t been \n cleaned or worked on to any great degree. I believe it was outside \n for about two years prior to them building the hangar for it, and \n it had been rained on. There were many ports of entries for water, \n and so water had pooled inside the bottom of the fuselage and it had \n left a lot of staining from dirt, waterlines, etc. and also had caused \n some minor pitting and corrosion. Nothing that would weaken the structure \n of the aircraft but certainly had caused some deterioration.\n Ross-Nazzal: \n Tell us about that move, when you decided to move her from that hangar \n into this facility.\n Mautner: \n Until I walked in March 1st of 2004, I had never touched Enterprise \n and I had really nothing to do with it. I recall being in the hangar \n the day they moved it in, and just about all the employees who were \n out here took a break to watch them tug it down the tow way from the \n airport and into the building.\n What was noteworthy at the time was that they had cut a notch in the \n back wall of the space hangar in order to permit the vertical [stabilizer] \n to come through. It was the only way to get Enterprise into that building \n without taking the vertical off, which would have been a prohibitively \n difficult task. I came back the next day and the blanking plate had \n been restored as if the building had never changed at all, so that \n was pretty neat. It was a well-prepared move.\n That took place in the early fall of 2003, and while I was aware that \n it was in the building, it was sometime after January 2004 that I \n was notified that I would be working on it in preparation for the \n opening of the space hall [James S. McDonnell Space Hangar].\n Ross-Nazzal: \n What was your expertise prior to working on Enterprise? Were you working \n on aircraft restoration?\n Mautner: \n My background is—the British would describe it as checkered. \n I did a whole lot of things before I got directly involved in airplanes. \n I started out as a kid building model airplanes, and I never gave \n that up. I always built model airplanes, and I continue to do so. \n And I was always a museum rat, an airplane museum rat. Whenever we \n traveled, wherever we went, there was always a stop at an airplane \n museum somewhere.\n Somewhere around the late 80s, my wife and I had moved our children \n to southern California, and I began to volunteer in some aircraft \n museums out there. Planes of Fame in [Air Museum] Chino, and later \n at the Command Museum at U.S. Marine Corps Air Station El Toro, which \n was right in the town I lived in so it was convenient to work there. \n I started doing structures work.\n In January of 1989 I left a former career and made a great midlife \n career change whereby I wanted to work in an aircraft museum. I had \n enjoyed volunteering on Saturdays so much, I went to Cal [California] \n State University, San Bernardino, for a Museum Studies certificate \n program. My background was in auto mechanics, and I needed to create \n a resume to get into working on aircraft, especially in museums. I \n enjoyed it immensely but had a problem with the hours of class.\n By great good fortune, McDonnell Douglas [Corporation] was hiring \n mechanics in the summer of 1989. Actually they’d been hiring \n prior to that, but that’s when I found out. I went and I applied, \n and they hired me immediately because of my automotive experience. \n I became a structures mechanic on the C-17 [aircraft] program in Long \n Beach, California. That was my first real hands-on experience with \n real aircraft manufacturing.\n I worked at McDonnell Douglas from August of 1989 until June of 1994, \n when there were massive layoffs. They were losing their contracts \n for MD-80s [airliners], and the MD-11 [airliner] never took off. The \n C-17 was in a state of flux because Congress and the Air Force were \n at loggerheads, so I and tens of thousands of others were laid off \n during that year.\n Meanwhile I had gone from my certificate program at Cal State, San \n Bernardino, to a Master’s program at Cal State, Dominguez Hills \n in Historic Preservation. Because John [F.] McDonnell of the McDonnell \n Douglas Corporation was interested in an educated workforce, they \n paid for almost all of my Master’s program, everything but my \n research courses and my thesis. \n I had early on decided that my thesis was going to be in aircraft \n preservation at the National Air and Space Museum. I thought that \n they were the best. They were the ones that were writing about aircraft \n preservation, and I really wanted to write about how they did it, \n because that seemed to be the benchmark.\n In June of 1994 I was laid off, and I immediately contacted the museum \n to do an internship back here so that I could do further research \n for my thesis. In August and September of ’94 I did a 60-day \n volunteer internship. I was outside of their regular program, so there \n was no stipend; I just came in and volunteered. I worked in the restoration \n shop during the day until about 3:30 [p.m.]. I’d then go across \n the river, downtown to the main museum and I would work in the archives.\n The people in the archives—I don’t know why, I was a total \n stranger to them—they let me stay after hours. In fact, at five \n o’clock when they left, they issued me instructions on what \n lights to turn off and how to shut down the computers. I would stay \n there till 8:30 so I’d get all my research. I had one other \n paper and I had my thesis to do, so I spent almost every weekday night \n at the museum doing that.\n They said, “Stay in touch; we’re going to be hiring.” \n The hiring was going to ramp up for this facility, this new Udvar-Hazy \n Center, so I stayed in touch. Every month I contacted somebody associated \n with the restoration facility, and finally in 1997 they said, “Something’s \n going to happen. Get back here; be nearby.” So I stayed with \n relatives and I hung out and I volunteered, and finally in February \n of 1998 they hired me. They did it because I had the academic background, \n but I also had, from McDonnell Douglas and my volunteer efforts, the \n mechanical background as well. I was very fortunate that the timing \n worked out. \n My background is chiefly in aircraft structures, which would be the \n main metal pieces that form the basis of the aircraft upon which everything \n else is hung. After I get through with my work, then the hydraulics \n people and the fuel people and the electrical people and the power \n plant people take over.\n Ross-Nazzal: \n What sort of planes have you worked on since you began working here?\n Mautner: \n The first one that I started working on was a Japanese single-engine \n floatplane bomber, [the Aichi M6A1 Seiran]. It looks more like a fighter \n with floats, but it was in fact a strategic bomber that the Japanese \n designed to fold up into an 11-foot diameter circle and fit into the \n tube on a submarine. This submarine was an aircraft carrier, literally, \n and it carried three of these. Very significant aircraft, because \n it was designed to bomb the Panama Canal locks. \n The submarine was probably more technologically advanced than the \n aircraft. It was designed to go 27,000 miles without refueling, and \n they actually did have plans to use these small bombers to bomb New \n York [City, New York] and possibly even Boston [Massachusetts]. It \n was a metal airplane, and my sheet metal experience came to good use \n there. \n After that, they put me on a World War I bomber, a twin-engine fabric-and-wood \n bomber. That airplane was a Caudron G.4, a French bomber that the \n United States tested but never purchased. Because we had no combat \n aircraft in World War I, we had to go to the French and the British \n and the Italians. That was an excellent experience because that broadened \n out my sheet metal and structures experience to wood and fabric. That \n airplane is also currently on display here.\n The next aircraft is one I’m still working on. We have a limited \n number of people here, so we get shifted off of whatever project we’re \n on to take care of other things in the museum, such as creating a \n display or working on the Enterprise for nine months in 2004. From \n about 2000 until this year, I’m still working on a World War \n II German night fighter, the Heinkel 219, a huge metal twin-engine \n aircraft, where again my airframe and sheet metal experience has come \n into use.\n Ross-Nazzal: \n Tell us what lessons you learned from working on those aircraft that \n you applied to the Enterprise.\n Mautner: \n In my case, precious few. It was a great departure from what we call \n restoration, where you actually have to fabricate or replicate or \n repair structural pieces of an aircraft. This was largely a massive \n clean-up and painting effort, so it falls under the role of preservation. \n That’s really our role here, preservation. Everybody talks about \n restoration, but restoration is the court of last resort when everything \n has pretty much gone to pot and needs to be drastically repaired or \n replaced.\n What we did on Enterprise fell under the heading of preservation, \n but we called it a refurbishment because we didn’t do anything \n to repair it. It was still considered to be NASA’s aircraft, \n and it came to us in such good structural condition that there really \n was nothing to repair on it except for the paint work.\n Ross-Nazzal: \n Would you tell us about working with NASA and USA [United Space Alliance] \n on this project? I understand they were involved.\n Mautner: \n Yes, they were. When we first began on March 1, 2004, we were pretty \n much on our own. There were just three of us: myself, Anne [C.] McCombs, \n and Steve Kautner. We walked into this big empty hangar. The only \n thing that was in it at the time was Enterprise, and it didn’t \n even have a barrier around it because the public wasn’t allowed \n in that building yet. We just kind of stood there and wondered, “Where \n the heck are we going to start on this thing?” \n We had no high-lift equipment—all we had access to were ladders—so \n we decided to start working on the landing gear. We had no contact \n with NASA outside of certain parameters that they gave us in cleaning \n and painting it, that we were given by the curator, Valerie Neal. \n Without anything else but ourselves, we began the cleaning process \n at the lowest part of the Space Shuttle, areas that were accessible \n initially to us.\n Our experience with NASA and USA came a little bit later, and [their] \n big project was the removal of all of the cockpit windscreens, the \n front ones. I believe that there were three panels in them, and there \n were some engineering discussions as to whether they could get away \n with just two panels. The challenge for us was to provide them with \n the platforms to get up there and work off of. \n Anne and I were involved, and at that point we had a lot of mechanics \n from United Airlines [Inc.] come in to assist us. Their bosses permitted \n them to give us some high-lift platforms that were attached to trucks. \n We could maneuver a truck in and then raise a platform and extend \n it out over the nose of Enterprise in order for the USA guys to remove \n those windshields. The other problem was these windscreens were very, \n very heavy. They required quite a bit of effort to crane off of the \n Shuttle.\n At that point, I began to get to meet some of the guys, most specifically \n from USA. One of them was a fellow named Tom [Thomas W.] Roberts, \n and Tom’s my friend now. He was back here for the last visit. \n There were several other mechanics that came up from KSC [NASA Kennedy \n Space Center, Florida] who I would count as really good acquaintances, \n friends, who we worked with. Got to meet Kevin [C.] Templin. He was \n always there, very personable fellow, and always a go-to guy for information \n and leadership. \n Whenever they came back, we dropped what we were doing and became \n assistants to them, to the point where in this last visit we were \n almost considered more one of their group than as a separate entity, \n which was kind of neat. The things I saw from everybody in NASA and \n USA was enthusiasm, expertise, positive attitude, and great great \n teamwork. The first thing we noticed is how well they worked as a \n team. \n Many times we didn’t know that these guys had never worked together \n before, and yet it was a seamless effort. You got to talking to them, \n you got to know them, and you realized, this guy never worked on the \n back end of the aircraft. He always worked on the front and didn’t \n even know the guys that they were working with. That was inspiring \n to us to see that kind of effort that they used to put in.\n Some of my recollection of my efforts with NASA and USA get a little \n bit confused, because some of their efforts were during that nine-month \n period when we worked on preparing Enterprise for the opening of the \n space hangar, but several of them came later. They would ask Anne \n or myself or others from the previous team to come back and assist \n them. I believe that the windshield effort was made during that nine \n months we were there, but our other efforts with them came later, \n so they’d be outside the parameters of my work on the Enterprise.\n Ross-Nazzal: \n I also read that you had to replace the OMS [orbital maneuvering system] \n pods because they had rotted.\n Mautner: \n Yes, we did. One of the first things we did was to remove those. Tony \n [Anthony W.] Carp was the team lead for the lifting of the OMS pods. \n They were a large aluminum plate that mated up to the surface of the \n Shuttle, but everything above that aluminum plate was basically wood. \n Plywood flooring, then 2 x 4 and 4 x 4 framework, and then they put \n a fiberglass cover over the top of that.\n We discovered how weak the structure was when we started to tighten \n up the fittings for the lifting rings. We’d gotten a guy inside \n to tighten the nuts up inside because they seemed loose, and he kept \n tightening and tightening and tightening. We suddenly realized that \n wood was so soft from moisture that he was just tightening the nut \n and the washer right into the wood. We had to be very, very careful \n lifting those OMS pods off.\n The OMS pods were never planned to be on the Shuttle when the space \n hall opened. That was something that we were going to install later, \n so our job now was to focus just on the Shuttle, and guys back in \n the shop worked on the OMS pods.\n Ross-Nazzal: \n Are they on there now, have they been replaced?\n Mautner: \n They were put on I think in 2005, 2006 time range, and before the \n last visit to get Enterprise ready we removed them. They’re \n out in the shop, on the floor. The guys from USA put the ALTA [Approach \n and Landing Test Article] pods on. Those are the travel pods. They \n look like OMS pods, but they’re real metal. They’re real \n structural, flightworthy, and the tail cone fits up to those components.\n Ross-Nazzal: \n Tell us about cleaning and painting the vehicle. That sounded like \n a very large chore.\n Mautner: \n Yes, it really was. I started on March 1st to clean the right main \n landing gear bay and ultimately the landing gear and tires. Steve \n Kautner took the left bay and did the same, and Anne McCombs worked \n on the nose gear and the nose gear bay. It wasn’t until March \n 22nd that Tony [Carp] came on the team, and his job was to clean the \n entire vertical surface with the ultimate purpose of coring or filling \n the holes that the woodpeckers had made.\n Somebody might at this point ask, why would woodpeckers go after polyurethane \n foam? The explanation we got—I’m not an ornithologist—is \n that woodpeckers make the noise they do as a mating call, not just \n to get to bugs inside of the tree. Once they got through that polyurethane \n foam, they were banging on an aluminum surface. It made quite a noise, \n and it was far-reaching. There were about 75 holes ranging from very \n small, smaller than your pinky, to much bigger than your thumb. \n We were using, on NASA’s recommendation, Amway LOC [liquid organic \n cleaner] cut, one part LOC to ten parts water, to wash everything. \n It’s a mild organic detergent, and it’s biodegradable. \n This was what USA used, so this was what we used. And this is where, \n in the wheel wells, I found most of the hornet nests. Once you got \n the water into them they just started running down the beautiful epoxy \n white paint panels inside.\n At that point, once Steve had finished his wheel well, he started \n working on the bottom. He started up at the nose where he could reach, \n because you couldn’t get a ladder under there. He’s all \n scrunched up, just scrubbing away—and he was a bear of a guy. \n He had a lot of strength, and he just kept working down the bottom \n of the aircraft, all of the black-painted area of the aircraft. \n At that point we started using some lift equipment, JLG [Industries, \n Inc.] lifts and scissor lifts, to get up on the wing. We began cleaning \n the wing while Tony is cleaning the vertical, and we did this day \n after day after day. Then sometime in the summer, I believe in July, \n we began prepping the center fuselage for paint. The center fuselage \n does not have polyurethane foam on it, and that center fuselage extends \n from just behind the cockpit to back underneath the OMS pods. We also \n had to clean the payload bay doors. At this point we finally were \n able to get scaffolding in. We had a local company contracted to put \n scaffolding all up the sides and then bridge the top of the Shuttle. \n \n This is where things kind of got interesting. Initially it was doing \n just a lot of sanding on the side. Apparently the aircraft had been \n painted with aerospace quality polyurethane or epoxy paint and primer, \n but when it went on tour in 1984 it had started to look a little bit \n secondhand. They used a latex house paint to give it a quick repaint \n to make it appear better at the [New Orleans, Louisiana] World’s \n Fair here in the United States, and then they went to the Paris Air \n Show with it. This was all prior to delivering it to us.\n This paint over the years had become very brittle and very hard and \n would be quite a challenge to remove. Taking it off the sides was \n pretty easy. We were able to use electric sanders or pneumatic sanding \n devices, hand-held devices, and literally sand all of that down to \n the original paint. The sanding gave the surface a real good surface \n for paint to adhere to once we finished it and cleaned it up, but \n the real challenge came with the payload bay doors. We were instructed \n by NASA to not use any metal tools, any caustic chemicals, or any \n abrasives to remove the paint.\n We got up there, and they thought it would be really easy because \n a lot of the paint on the payload bay doors had flaked off, and some \n of it was literally sitting up. They said, “Just use duct tape \n and press it down real hard and pull it off, and that paint will pull \n up.” Well, that didn’t work at all. \n All of our communications with them were through Valerie Neal, the \n curator. The other thing we weren’t supposed to use was heat, \n so we went to Valerie and said, “The duct tape just isn’t \n cutting it.” They said, “Use plexiglas scrapers.” \n Plexiglas is a real hard plastic. We would cut up sections of it, \n rectangles of it, and then make a sharp edge at one end with a grinding \n wheel. That wasn’t cutting it either.\n One day I’m sitting on the scaffolding, trying to get this stuff \n off, and I see to my left this movement from about chest height over \n the curvature of the door. I look over, and it’s Steve Kautner \n with a metal scraper and a heat gun. I’m going, “Steve, \n what are you doing?” He goes, “This is the only way it’s \n coming off.” I said, “Okay, stop. Let’s talk about \n this.” \n We went back to our conservator, and I asked him, “Do you have \n a surface temperature gauge?” We went back and attached this \n temperature surface gauge to the area that we were putting the heat \n on, and we got some heat parameters. We were getting heat in the range \n of about 175 degrees [Fahrenheit] if we kept the heat gun moving, \n so we went back to NASA and they said, “Okay, you can do that, \n but no temperatures over 200,” or 250.\n All of a sudden this became easy, but we now found out very quickly \n why they didn’t want heat up there. The doors are made of carbon \n graphite, or carbon–carbon. It’s skinned with an almost \n aluminum-foil thickness coating of aluminum. It’s not like sheet \n metal, it’s more like a foil, and if you left the heat on there \n too much it would blister. We got a couple of thumb-sized blisters; \n we damaged the door. This concerned us, and we kept talking to each \n other. We’d just make sure we’d keep that heat gun moving \n and don’t overheat one spot.\n It became very evident very quickly why they didn’t want heat \n up there. They had done the same thing at some time in the past, only \n their blisters were much bigger than ours. That told us a) it’s \n been done this way before, and b) there’s a repair for it. We \n did make a few more blisters, an inevitable result of what we were \n doing, but we knew that if they needed those doors, they could be \n made flightworthy. \n Then, because there was still a lot of paint residue clinging to the \n surface, we got down and used acetone. We were all garbed up with \n double-canister air masks. There’s terrific pictures of us all \n down on our hands and knees on the door, all tied off because we’re \n high up on the top of the Shuttle, scrubbing with Scotch-Brite scrubbers \n and acetone to get the surface completely clean and ready for paint.\n That was another thing they told us, “You can’t stand \n on the doors.” So we had these four bridges across, and we’re \n all leaning off these bridges and working off of them. Again, Steve \n Kautner’s the first one to step off right on the doors. There’s \n no other way to get out there, there was no way. I said, “Okay, \n stop. Let’s talk this over.” We went down inside the Shuttle, \n and we looked up at the doors and we could see the latching system. \n \n We knew that those doors in zero gravity could open without any support, \n but we knew that they had big supports on them when they opened them \n in the hangar down in KSC. We looked at those supports, and we looked \n at how the doors fit together. It’s built like a bridge as long \n as they’re closed and locked, so four, five, six of us all were \n out on our hands and knees on the doors, scrubbing away to get the \n last residue and make a really good surface for paint.\n That brings up another aspect for me. I couldn’t wait every \n single day to come in. I’m not a space guy, I’m an airplane \n guy. In truth, the Shuttle is probably more like an aircraft than \n a spacecraft, and that one [Enterprise] never did go into space so \n it really was just an aircraft. I never followed space, I never got \n excited about it, but I got excited by this project. I couldn’t \n wait to get in every single day. Every day was interesting and exciting, \n and we were making progress—slow—but we were making progress. \n \n When they put the scaffolding up in July, we now had to climb this \n scaffolding. We had our harnesses on and we had our tethers, but we \n couldn’t tether off anywhere on the way up. We’re climbing \n 30, 35, almost 40 feet in the air every morning, every break time, \n every time you had to leave, and then when we got up there we would \n tie off and secure ourselves. But that little element of danger made \n it exciting, and we talked about that a lot too.\n We had two injuries, but only one on Enterprise. One of our guys did \n abrade his hand, but working on something else while we were out there, \n nothing to do with the Shuttle. One injury, very minor, on the Shuttle. \n One of our members fell off the payload bay doors, but he was tethered \n and it stopped him, and all he did was bang up his knee. I think we \n lost him one day, but that was it. The fact that we talked almost \n every day about the safety issue, that he was tethered off—we \n didn’t have more issues. \n The other thing that made it interesting and fun is that we were on \n this huge, very obvious artifact in the middle of this totally empty \n hall, and the public could come in to the front of the hall—there’s \n an overlook up above—and watch us. I think the element of being \n on stage made it exciting and interesting, too.\n There were some stresses. We always seemed short of personnel. At \n one point I realized there were a whole bunch of guys working on the \n OMS pods, and I had to convince my management that the OMS pods aren’t \n needed by October 19th, but the Shuttle is, so can I have more guys? \n And I got more guys.\n Once we had sanded down the sides and had removed the paint from the \n doors, it was time for a final prep and primer and paint. We had four \n guys do that, and we had a couple of issues that we had to deal with. \n One, it had to be an aerospace quality paint. PPG, Pittsburgh Paint \n and Glass, has always supplied us with our paint system that we’ve \n used exclusively for 20 years back at the [Paul E.] Garber [Preservation, \n Restoration, and Storage Facility, Suitland, Maryland], our restoration \n shop. Dave Wilson got in touch with them, and they turned him over \n to their aerospace division. They donated what amounted to about $15,000 \n worth of white polyurethane aerospace paint and primer.\n Our next challenge was we could not spray in the building because \n the building had an open port, and some of the ventilation systems \n functioned with the aircraft hangar. Also, the filters in the ventilation \n system would get clogged up with any paint residue and dust, so we \n had to either brush or roll it. Dave talked with the people from PPG, \n and they had an additive that allowed the paint to flow so that it \n would not appear to be rolled-out paint. It wouldn’t be textured, \n it would be nice and smooth.\n We got the paint on September 17th—keeping in mind that this \n hall is supposed to open on October 19th of 2004. They had to rest \n assured that all the fumes would have dissipated by 10 o’clock \n opening the next day, so we painted at night. It was Tony, Dave, myself, \n and a fellow by the name of Bob Weihrauch who did it. We would come \n in, and that hall was completely black except for the Klieg lights \n that we worked under. We rolled out the primer, and then we rolled \n out the white. [We rolled out two coats of primer and two coats of \n white.]\n I love working here at night because there’s no people here, \n the lighting is all dim, and it’s like going into a great cathedral \n at night when nobody’s around. It’s just beautiful. It’s \n quiet. You’ve got all these objects that you love, many of which \n you’ve worked on, and there’s just a neat thing about \n working here at night. We had a really good time. We had a really \n good crew. I don’t recollect ever having any arguments or disagreements \n or problems, we just really had a good time.\n One outstanding recollection is this facility is used by organizations—police \n organizations, FBI [Federal Bureau of Investigation], the MWAA, Metropolitan \n Washington Airport Authority. They have their own SWAT [special weapons \n and tactics] team and security forces. They do a lot of work in here \n at night, but we didn’t know that. One night three of us are \n up on this scaffolding, and Dave Wilson is on a scissor lift bringing \n paint up to us, and all the sudden there’s movement in the hangar. \n \n Keep in mind that outside of this cone of light that we’re in, \n everything’s dark out there, but there’s movement. We \n look down, and there’s 20 or 30 guys in black helmets, black \n outfits carrying guns, and they’re swarming like ants through \n our hangar. Well, I immediately was not concerned because none of \n them are looking at us or pointing the gun at us. We’re in white \n bunny suits, paper coveralls, and the Klieg lights are on us. We’re \n out there, very obvious.\n One guy kind of moved up to Dave, and he goes, “Have you seen \n her?” And Dave goes, “Seen who?” He goes, “Have \n you seen her?” and then he had to move, because you’re \n not supposed to look for hints. Apparently they were on an exercise, \n and one of their supervisors came in two hours ahead of time and hid \n herself somewhere in the museum, and they had to find her. Nobody \n told us; our security had not advised us that this was going to happen. \n \n Another time, during the day, I was working on the Shuttle and nobody \n else was around. Security came up to me and said, “We have a \n congressman or a senator here with his daughter back from college, \n and he’s got his office manager and his security detail. Would \n you come down and talk to them?” So I walk inside of this circle \n of Secret Service people, and they’ve got their radios and they \n all look very, very official, and I started to talk to them. I said, \n “What state do you represent, sir?” He goes, “Oh, \n one of your neighbors.” Meanwhile, his handler, his office guy, \n did all the talking. \n I went back that night and I looked all over the Internet for our \n neighboring senators and congressmen, and I did not find him—because \n you can get the photos of all of them. I went back to security the \n next day and I said, “What was that all about?” He says, \n “Oh, it’s just a test.” It was Secret Service in \n a practice run, and these guys were just members of the Secret Service \n that were playing a role to make it seem realistic. There are a lot \n of activities that go on here that are very interesting. But again, \n I was not forewarned of that. \n The other two events that were really interesting—Joe [H.] Engle, \n one of the test pilots for Enterprise, came back and spent a day with \n us. He is just a prince of a guy, just a super guy. We got to tour \n him around and show him what we were doing, and he was just the friendliest \n guy and was very very grateful to us restoring “his” bird \n back to its original condition. A couple weeks later, we all got an \n 8 x 10 of him standing by Enterprise out at Palmdale [California] \n back in’77, ’78, with a beautiful handwritten note and \n one of the original patches from his missions. It was just neat, a \n neat thing. \n The other very very interesting visitor to us was Colonel Pamela Ann \n Melroy. She’s a two mission pilot and one mission command pilot \n [commander]. She was also on the Columbia [STS-107 accident] investigation \n team, and she led the cockpit reconstruction area. My recollection, \n she was about five-foot-five, five-foot-six. I immediately made a \n connection, because she was a test pilot on the C-17 program. As I \n had worked on it, we had some things to talk about. \n I got to tour her around, and the thing I recollect so well is that \n she made a comment when we were in the cockpit. The cockpit of Enterprise \n is for the most part empty. The dashboard is there, but there’s \n only about three instruments, and they’re all old analog instruments. \n The seats are not there. She looked at that dashboard and marveled \n at the fact that it was the first time she’d seen one in a long \n time that wasn’t bits and pieces, because she had been looking \n and reviewing and trying to reassemble pieces of Columbia’s \n instrument panel.\n This diminutive colonel in the Air Force—always at the end the \n photographer comes in and takes a picture of us with the VIP [very \n important person]. Anne and I were there at that time, and we’re \n all standing off to the side, a foot away or something. She says, \n “Hey, we’ve been together all day. Come here!” She \n pulled us in right next to her to take the picture. We also got an \n 8x10 from her, signed, with a thank you and mission patches. Those \n are two of my favorite treasures from that whole time there. \n Ross-Nazzal: \n Nice recollections of that time period. \n Mautner: \n Yes, it really was. We got it done. Everything seemed late—getting \n personnel, the scaffolding came late, the paint came late—but \n we got it done. We got it done on October 18th, and October 19th was \n the day the media and the museum VIPs and everybody showed up to see \n it. It was a moment of pride because it really looked good. \n Tony and I stayed after October 19th. Tony had some more work up on \n the vertical to do, and the one area that we hadn’t really addressed \n was the propulsion bay. I would work in the propulsion bay; he’d \n work up top. We did this at night because the gallery was open to \n the public, so we did a lot of night work from October until we went \n away on Christmas vacation.\n Most of cleanup in the propulsion bay—there was some FOD [Foreign \n Object Debris], there was old hardware. It went back to when it was \n delivered to us. There were some sections of insulating aluminum, \n nuts, bolts, washers, things like that. A lot of water had been back \n there, and the floor panel structure was a huge, thick piece of aluminum \n that they had milled out to look almost like a waffle. The floor looked \n like a large waffle pattern, so there’s all these thousands \n of little pockets that had held water, and I had to clean them.\n We had some nights where we had events, and the events usually allowed \n people to roam into the space hangar, so I would take my knapsack \n and my coffee and my lunch, my snacks, up into the propulsion bay. \n Something you don’t normally do around artifacts, but I didn’t \n think I could come out because of the events they had going on. I’d \n stay up there for eight hours. Well, there’d be a potty break.\n We finished before our Christmas break, and then both of us reported \n back to the Garber Facility the first week in January 2005. It was \n rewarding every day, but they also rewarded our team with the Museum’s \n Peer Award. The [museum] gives two Peer Awards every year, and the \n Enterprise Preservation Team was awarded [one of them] for 2004. It \n was a great experience.\n Ross-Nazzal: \n What are you working on now that Enterprise is getting ready to take \n a new journey in its history to go to the Intrepid [Sea, Air & \n Space Museum in New York City]? Are you working on that?\n Mautner: \n I did work on it. I don’t know what my role will be in April \n when they come back, but the whole month of February I was assigned \n out here, along with Anne and one or two other folks, to escort NASA, \n USA, and some folks from [the] Boeing [Company]. “Escort” \n has a kind of static sound to it. We were essentially here to open \n doors that they couldn’t get into, including getting to the \n restrooms, and we were able to get them discounts in our shop and \n at McDonalds. \n And then pretty soon it was, “Oh my gosh. We don’t have \n this; do you have this tool?” So we’re running for tools. \n Later, as our professional relationship evolved, we were forklifting \n equipment for them. They had forklifts, but we were able to help them \n with that. We had a JLG man lift that we were able to take them and \n some of their equipment up to different places on the Shuttle, and \n also on the tail cone, which they assembled right out here in this \n shop. As to my role when they come back on the 19th, I have not been \n told what that is yet. I hope [I] do, I have some friends I’d \n like to hang out with again.\n Ross-Nazzal: \n Are you preparing in any way to accept [OV-103] Discovery, or are \n you primarily working with the Enterprise?\n Mautner: \n I’m not. My responsibility is to work on this German night fighter. \n We still have a wing to complete. The airframe fuselage is out in \n the museum right here now, but the wing has to be done and a lot of \n other details. Between escort duties and whatnot, I’m working \n on that. We’re told that with everything else that’s going \n on, we’re going to be moving our shop from Maryland out here. \n I don’t know how that’s going to work out with the Discovery–Enterprise \n exchange, that’s yet to be determined.\n Ross-Nazzal: \n It must have been rewarding to receive that from your peers, that’s \n wonderful.\n Mautner: \n Very, yes. I think Valerie Neal, former NASA and our curator, was \n probably instrumental in getting that for us. It was pretty neat.\n Ross-Nazzal: \n Rebecca, did you have any questions for Ed?\n Wright: \n Just one, and I think you might have already answered it. Did you \n ever come back and do anything else with Enterprise?\n Mautner: \n I have once or twice, but it was generally just escort duty. Those \n were times where it was just sit around and wait for whatever the \n folks, predominately USA, needed. I did come back, and we did reinstall \n the OMS pods. That was pretty much the original team again that did \n that, and we brought a couple of other people in to assist because \n you need spotters and tag line holders. \n Wright: \n You mentioned that you would escort for whatever they needed. What \n were the types of things that they would be coming back for?\n Mautner: \n Every time USA came here, they had a lot of supplies. Rather than \n transport them back, they would leave them out and we would store \n them in our barn, which is out in the back corner of the property. \n So the first thing we’d have to do is get their supplies, and \n that ran anywhere from tape to paints to epoxy glues.\n Recently, within the past year, they came back and we took them up \n on a JLG man lift. They did the work, and I would just maneuver it \n around to a position that helped them. Like I said, “escort” \n sounds very static, and sometimes it is, but a lot of times there’s \n a dynamic component, which is the most fun of all. That’s how \n you get to know these guys and get to see what they do.\n Wright: \n Were they doing types of restoration themselves?\n Mautner: \n Yes, about last April they came back to begin the process of making \n Enterprise airworthy. That involved making an assessment of all systems, \n and they also did the [landing gear] retracts. The main job was to \n get the hydraulic system working so they could retract the landing \n gear. Part of our position as escort was to shut down areas of that \n hall to keep the public away, because the machine, what they call \n the mule, fires up the hydraulics system, makes a lot of noise. Also \n there’s part of the retract that there’s noise involved \n as well.\n We did a lot of work stanchioning off. Moving stanchions back, and \n then when it was all done, moving the stanchions closer. We were trying \n to do this juggle between making sure people were safe and USA had \n enough room to work in, and also still keeping the hall open to the \n public. Our whole purpose is to let them get as close to this stuff \n as we can.\n There were a couple of times where, yes, it was pretty static, but \n other times we were pretty involved in helping them. And everything \n back here is secured. You can’t get into the restrooms without \n our badge, so we were escorting to the restrooms. Eventually, and \n especially in this last trip in February where they did the final \n preparation for flight, the security office provided them with badges \n that allowed them to open all the things they had to get into. They \n had the same access employees had. \n I think that speaks of the type of relationship that we’ve always \n felt we had with NASA, that they weren’t an alien organization \n that comes in and invades our space but rather cohorts, colleagues \n that we feel—not exactly one with, our roles are different—but \n certainly very good friends with.\n Wright: \n Is this a very unique exhibit here compared to all the rest? Do you \n have any other kind of associations with other groups that come in \n and work on their planes?\n Mautner: \n No, there’s no other organization that has the relationship \n that NASA has with us. I think that’s historically because we’ve \n been the repository of first choice for NASA, but also because Valerie \n Neal, who came from NASA, has maintained a very, very good relationship \n with all the leadership there. It’s been a very collegial relationship \n from the very beginning. Because of that, your relationship personally \n to these people is much more relaxed, so it’s much much easier \n to make friends. I have a couple buddies that I email once in a while \n back at the Cape [Canaveral, Florida]. It’s a good relationship.\n Ross-Nazzal: \n Any other last thoughts? \n Mautner: \n I think I’ve covered the important aspects of my relationship \n with the Shuttle Enterprise and with the folks from NASA and USA. \n We had a good team. I think it was kind of neat because management \n left us alone; they left us to make a lot of our own decisions. Everybody \n worked with the goal in mind, and we reached the goal. We finished \n Enterprise, it really looks beautiful. It took us nine months to get \n it in the shape that it’s in right now, and I have to say I \n am quite proud of how it came out. \n I do recollect one of the stories that I’ll tell you. When we \n began to strip all the paint off, we traced all of the markings—the \n NASA worm, the “United States” on the side, the “Enterprise” \n itself—so that after we painted it we could restore those. We \n noted and photographed the fact that these had been hand-brushed. \n Somebody had done a stencil, and then they hand-brushed in the black \n color. As we began to strip the aircraft, we realized that there were \n other markings, other NASA worms and another “Enterprise” \n and another “United States” underneath that exterior coat \n of latex house paint. \n We traced those as well, and at that point the curator had to make \n a decision. Are we going to restore it to when it was originally painted \n and lettered, or are we going to paint it as it was presented to us \n in 1985? The decision was made to paint it as it was delivered to \n us. Those letters were all in black, and the letters we found underneath \n were in lighter shades of dark gray. \n At the time that we traced them back on the aircraft, the question \n was asked, “Well, who feels comfortable painting them back on?” \n I mean, none of us are professional sign painters. Out of the five \n of us there, four chose to do it. So what you’ll see out there \n is our hand-lettered letters, and they are letter perfect. They came \n out really, really well. Three employees and one volunteer painted \n them all in. That too raised our level of pride and involvement and \n attachment to the Enterprise. \n I have mixed feelings about it going. On the one hand, it’s \n very important for us to have an orbiter, and Discovery is a much \n more significant vehicle to have in our collection, so I’m happy \n about that. On the other hand, all of our handwork is being sent off \n to somebody else. I’m not going to lose sleep over it, but I \n will think, how are they taking care of it? Is that beautiful paint \n job still squeaky clean and nice, and are all those tiles in good \n condition? So yes, I am concerned to some degree at it going off to \n other hands.\n Ross-Nazzal: \n Any plans to go to New York and see her once she’s in her permanent \n location?\n Mautner: \n No plans at this time, no.\n Wright: \n And will you be working on Discovery when it comes?\n Mautner: \n We’re not going to touch Discovery. This is another aspect that’s \n pretty neat about getting Discovery. The ideal airplane or spacecraft \n to get into our collection is the one that just went on its last mission. \n If we could get a Boeing B-17 [aircraft] that flew a mission from \n England in August or September of 1944, and then was transferred right \n back into our hall out front, that’s the idea. You don’t \n want to touch it. You want to mount it in such fashion that you preserve \n the tires and take any other stresses out of it, but it would be great \n if it had the gunpowder stains and the engine stains and the oil stains \n and the rough-hewn patches that the mechanics put on the planes back \n in those days. \n And that’s how Discovery is coming to us. They haven’t \n cleaned it, outside of taking out the toxic materials in all the tanks \n and systems, so it’s not going to look like Enterprise at all. \n In fact, one of the people from NASA commented that Enterprise looked \n like a toy. I think what she meant was it looked like a model, and \n Discovery looks like a real orbiter. So we’re happy about that.\n Wright: \n Well, thank you.\n Ross-Nazzal: \n Thank you for spending some time with us this morning. We appreciate \n it.\n Mautner: \n You’re very welcome. It’s my pleasure to do it.\n [End \n of interview]\n \n \n Return \n to JSC Oral History Website"],["topImageUrl","https://www.jsc.nasa.gov/history/oral_histories/head_images/sc.gif"],["error",null]]}]}],["text","美國太空總署員工表示，\n他們是使用太空總署建議的\n【安麗 LOC】以「1:10」濃度\n拿來清洗太空梭及所有東西。\n\n這就是表示該產品是【極度安全】的！\n\n這樣，還有什麼安麗產品是需要擔心呢？\n\n\nhttp://www.jsc.nasa.gov/history/oral_histories/NASA_HQ/Experts/MautnerEM/MautnerEM_3-22-12.htm"],["id","5484461227031-rumor"],["createdAt","2017-01-11T01:17:00.000Z"]]},{"__iterable":"Map","data":[["replyCount",1],["references",{"__iterable":"List","data":[{"__iterable":"Map","data":[["type","LINE"]]}]}],["replyRequestCount",1],["user",{"__iterable":"Map","data":[["id","j4S8C_edDdU96oZupq3Xms9tKo2siS2uA7HMWfNP6BYlD1P_g"],["name","Oo好學上進的下營克莉絲汀oO"],["avatarUrl",null],["level",0]]}],["hyperlinks",{"__iterable":"List","data":[{"__iterable":"Map","data":[["title","キャンペーンに賛同！"],["url","https://www.change.org/p/request-permanent-court-of-arbitration-to-investigate-unlawful-use-of-force-of-hong-kong-police?utm_content=cl_sharecopy_17005953_en-GB%3Av5&recruiter=90670311&recruited_by_id=0040d850-5bec-43c4-8e81-c03a3953c79d&utm_source=share_petition&utm_medium=copylink&utm_campaign=psf_combo_share_message&utm_term=psf_combo_share_initial&share_bandit_exp=message-17005953-en-GB&share_bandit_var=v0"],["summary","(Update on 11/11/2019)\nURGENT! \nHong Kong need help from the World ! Please sign and support ! \nWe are writing to request Permanent Court of Arbitration (PCA) to formally Investigate Hong Kong Police for Unlawful Use of Force against Citizens since 9 June 2019\nAttn: Mr. Hugo H. Siblesz (Secretary-General of Permanent Court of Arbitration)\nDear Mr. Hugo Siblesz,\nWe are Hong Kong citizens. Hong Kong urgently need helps now!\nWe are writing to ask for your help to formally investigate police brutality against citizens of Hong Kong in the recent protests since 9 June 2019. A group of international experts appointed to help Hong Kong’s police watchdog conduct a study into the ongoing protests has said the body lacks power and urged for a more comprehensive investigation by an independent group with requisite powers.https://www.facebook.com/800660560021071/posts/2581953451891764/\nThe Hong Kong police front line police is getting insane. After the death of a 22-year-old student who died near a police clearance operation of a protest on last Friday. On 10 November, multiple arrests being made in Festival Walks, a shopping mall, and chaos scattered everywhere on the streets and in different housing estates.\n11 November 2019 is absolutely chaos. It is reported that the real guns shot by the police officers in different public areas.\nA Hong Kong police officer has shot a black-clad protester in the stomach at close range :\nhttps://www.facebook.com/800660560021071/posts/2583690221718087/https://www.facebook.com/800660560021071/posts/2583667861720323/\nA Hong Kong police motorcyclist appears to \"ram\" a crowd of black-clad protesters on 11 November:https://www.facebook.com/800660560021071/posts/2583726308381145/\nHundreds of thousands of people in Hong Kong took to the street in recent days to protest amid the controversial Extradition Bill. In the latest rounds of the protest, the Hong Kong Police Force (HKPF) has been using excessive level of violence and firearms against citizens who participated in peaceful protests. Confirmed crowd control weapons and ammunition in use including batons, pepper spray, rubber bullets, sponge grenades, live bullets, water cannon, tear gas. The sole purpose of this level of extreme violence “intent to injure”. It is believed to be a direct and flagrant violation the “UN Principles on the Use of Force and Firearms by Law Enforcement Officials.” Many unarmed protesters have been reportedly beaten or shot, some directly in their heads.\nSince 9 June, we believe that Hong Kong Police Force had committed the following crimes:\n1) Crime of aggression2) Conventional War Crimes3) Crimes against Humanities\nHong Kong’s anti-government protesters have sustained for five months. Here are some figures:\n1) It is 156 days of unrest since June 9\n2) 3,300+ protest-related are currently arrested, 10+ people are dead\n3) 6,200+ tear gas canisters used\n4) 15 bullet+, 2,000+ rubber bullets, 670+ foam bullets and 450+ beanbags bullets were fired\nRecord of the Hong Kong Police Brutality are as Follow:\n(A) Hong Kong Police Force Investigation Reporthttp://hkpfreport.org/\n(B) Hong Kong Police Violence Database:http://tl.hkrev.info/en/police-timeline/\n(C) Unlawful use of force by Hong Kong Police https://www.amnesty.org/en/documents/asa17/0576/2019/en/\n(D) Allegations of Hong Kong Police Force misconduct surrounding the 2019 Hong Kong protestshttps://en.m.wikipedia.org/wiki/Allegations_of_Hong_Kong_Police_Force_misconduct_surrounding_the_2019_Hong_Kong_protests\n(E) A Report of the 2019 Hong Kong Protest (written by Martin Purbrick, a former Royal Hong Kong Police officer whose service included Special Branch engaged in counter-terrorism intelligence)https://bit.ly/2nVuglA\n(F) The Six-monthly Report on Hong Kong - 1 January to 30 June 2019 (by Parliament Secretary of State for Foreign and Commonwealth Affairs)https://assets.publishing.service.gov.uk/government/uploads/system/uploads/attachment_data/file/843492/Hong_Kong_Six-monthly_Report_Jan-Jun19.pdf\n(G) Weapons of mass control, tactics of mass resistancehttps://www.reuters.com/investigates/special-report/hong-kong-protests-violence/\nFourteen Unlawful Use of Force by Hong Kong Police Force\n1) Shoot and attack both protesters and reportersUse live bullets dangerously. In October, a teenager was shot by wielding a pipe with live ammo at a point blank range with a revolver.https://youtu.be/PN4MvOrPotkAn Indonesia reporter was shot by police in rubber bullet on the face and become permanently blind in one eye.https://youtu.be/gWn5oRJKiM8https://www.bbc.com/news/world-asia-china-49910636\n2) Unlawful use of batonsPolice officers use batons to beat people, no matter they are protesters or un-involved bystanders. Police would keep beating people with batons while they lay on the ground with no resistance. That was inconsistent with the use of force guidelines that was taught in Police College.https://youtu.be/kW4A9hy3H3Y\n3) Improper use of riot control agents(RCA) - tear gas, pepper spray and water cannon. Use pepper spray on a person who posed no obvious threat is usual.https://youtu.be/4QvQTW40exYhttps://youtu.be/TAzkH4EncPA\n4) Arbitrary arrests people The arresting is from age 10 to 82. The police after failing to arrest the radical protesters, turned to arrest peaceful protesters, bystanders, young people in general and people wearing black clothes in young age, and even doctor.https://youtu.be/LnUFOjHYuxYhttps://youtu.be/yLbaXMaaOq4https://youtu.be/C_h1oJYyTe4\n5) Lack of visible identificationDuring many of the Hong Kong Police operations, the HKPF did not wear any ID badges nor did they produce any warrants. They had infringed the SOP (Standard Operating Procedure, and neglected the ROE (Rule of Engagement).https://www.hongkongfp.com/2019/08/23/tactical-squad-may-hiding-ids-order-protect-privacy-says-hong-kong-police-watchdog-vice-chair/\nhttps://www.scmp.com/news/hong-kong/law-and-crime/article/3017775/hong-kong-police-accused-provoking-protesters-and\n6) Restrictions on journalists and medicsJournalists:https://www.scmp.com/news/hong-kong/politics/article/3026940/press-associations-call-hong-kong-police-curb-attacks\nhttps://www.scmp.com/news/hong-kong/politics/article/3026940/press-associations-call-hong-kong-police-curb-attacks\nhttps://www.hongkongfp.com/2019/10/04/aggressive-obstructive-hong-kong-journalists-association-files-legal-challenge-police-treatment-press-protests/\nMedics:https://youtu.be/x_P0lBpgscs\nhttps://www.scmp.com/news/hong-kong/politics/article/3024947/hong-kong-police-apply-search-warrant-access-medical\n7) Brutal beatings, torture and sexual abuse protesters in police detention. Sexual Abuse in the police’s San Uk Ling Detention Centre (dark terror cell) which is located in an area close to the China-Hong Kong Border. It was reported that some students were tortured after arrest, leaving bone fractures. The police officers in the terror cell was “unnamed”, “unverified” and “unnumbered” and the cell has no cctv systems for recording what happened. The terror cell after the public decry is no longer used for detention. https://www.amnesty.org/en/latest/news/2019/09/hong-kong-arbitrary-arrests-brutal-beatings-and-torture-in-police-detention-revealed/?utm_source=TWITTER-IS&fbclid=IwAR06vFKbU5i_bRUaRwp0G_J8YvZK6zEudA8Zeiv6qdzDVMtZ-MObIgnVAH0\nhttps://mobile.twitter.com/thetimeusedtobe/status/1182292557595721730\n8) Raptor squad went undercover to target radical Hong Kong protesters These police disguised themselves as protesters. But in reality, they arrest vulnerable target like young girls and boys on the street in the name of action to arrest \"extremely violent rioters\". These target youngsters were bought into non-police vehicles by the undercover, then disappear without any trace and record.https://www.scmp.com/news/hong-kong/politics/article/3022457/elite-police-raptor-squad-went-undercover-target-radical\n9) Rumors of police kills peopleSince the occurrence of the 8.31 Prince Edward station incident, the \"police killing people\" rumors have been heard, and there have been many suspicious floating bodies, including Chen Yan-lin, a 15-year-old swimmer who had participated in the anti-Extradiction bill movement. The cause of death is unknown. Although the police denied twice, rumor is still spread. https://www.scmp.com/news/hong-kong/politics/article/3026439/hong-kong-protests-officials-attempt-sixth-time-debunk\n10) Inconsistent law enforcementhttps://www.hongkongfp.com/2019/07/26/hong-kong-police-breed-mistrust-uncertainty-selective-law-enforcement/\n11) Hong Kong becomes a police city In the name of law enforcement, the Hong Kong Police is allowed to extend their action of violence and brutality anywhere, anytime. Off-duty police officers were given telescopic batons and pepper spray for carrying and using when they were off duty.\n12) Spreading climate of fearThe Hong Kong Police Force has been accused of spreading a climate of fear. It has conducted hospital arrests, which had prompted protesters to become reluctant to go for treatment in public hospital, fearing they may get arrested by the police. Medical professionals may also be charged by the police for offering treatment to the wounded.The police banned protest and marches in various places in Hong Kong. The refusal to issue permits had eroded Hong Kong's freedom of demonstration. The police for rounding up various activists to frighten Hong Kong people into not protesting, even though these protests have been leaderless.Disregard the safety of the members of the public, police use unlawful force on peaceful or retreating protesters was usual. Hong Kong police can be seen in using excessive force against unarmed and non-resisting citizens and innocent commuters.\n13) Trash daily police press. Police lying to the public all the timeEveryday , it is anticipated that the Police Public Relations Bureau would be conducting another routine briefing at 4 pm Hong Kong time. It would not be surprising if they could find explanations, if not excuses, for the brutal force used and they would probably, if not certainly, put blames on the protesters for assembling illegally. It has also become a practice for the Police to condemn the protesters for violence while defending the police for whatever forces used during the operations, thereby damaging its own credibility.The Police has repeatedly denied accusations of tortures and sexual assault cases in detention center. The most notorious one was in San Uk Lang, which is proximate to the Chinese border. Victims had spoken both in public and online of the maltreatment that they have received and also what they have seen.In response, the police remained adamant that the accusations were false and demanded the victims to put away their mask and report to the police. One of the Deputy commissioner of police even issued a letter to fellow policemen, claiming that the accusation of “Black Cops” is non-existence and false.\n14) Without any conscious of damage cause by the unlawful and excessive use of force, Hong Kong police continues to consider its operation swell with unlimited power to enact laws.https://www.hongkongfp.com/2019/08/21/fall-hong-kong-police-force-new-branch-chinas-public-security-machinery/\nhttps://www.scmp.com/news/hong-kong/politics/article/3021258/beijing-stands-behind-hong-kong-police-state-media-while\nhttps://www.hongkongfp.com/2019/10/14/hong-kong-govt-considers-enlisting-staff-depts-become-police-special-constables-report/\nBeijing Stands Behind Hong Kong Police, Hong Kong People Need Help from the World\nEvery week, millions or thousands of citizens have come out to express their dissatisfaction towards the current political system and the performance of the government. Yet, the government chose to shift the responsibility of the political turmoil to the protesters and condemned violence selectively. They are turning a blind eye to the voice of the people, while pandering to the “white terror” and police brutality in Hong Kong.\nDespite the popular outcry from Hong Kong people from different sectors urging the Police Commissioner to issue an immediate official apology for its wilful attacks against citizens, people’s demand once again had not been heard. The reputation of HKPC is totally wrecked.\nAlthough we repeatedly demand the Independent Police Complaints Council (IPCC) to set up an independent inquiry to investigate HKPF’s misconduct and its gross abuse of power, once again the voice of the Hong Kong people is ignored. We seriously doubt of the independence, integrity and effectiveness of IPCC.\nMore peaceful rallies and protests will be ongoing held to openly condemn police excessive use of force as part of the ongoing anti-extradition bill protests. The Hong Kong people re-state their 5 major demands: \n(1) Independent inquiry on police brutality\n(2) Immediate release all protesters\n(3) Retract the characterisation of peaceful protests as “riots\" \n(4) Universal suffrage\n(5) Reform Hong Kong Police, disband the existing Police Forces\nWe are deeply worried that this unlawful use of force by the HKPF will persist. More unarmed peaceful protesters, mostly young people, will continued to be hurt and sustain a disproportionate level of injuries, some of which can be long term.\nWe seek for political settlement to end the violence that erupted following the anti-extradition bill in 2019. Human rights, political accountability and democracy should be introduced and put into the Hong Kong Police Ordinance. Police force is a major front on which these normative reforms were to take place. These reforms should seek to break the historical reliance of the political elite of Hong Kong/Mainland China on the police as an instrument of elite power and repression; to improve the account-and support the evolution of more a responsive, public-focused policing.\nWe implore the Permanent Court of Arbitration to take this crisis seriously and take immediate action to hold the HKPF to account. The Hong Kong government and the police force have completely failed to serve its people. It is only with the intervention of international organisations like the PCA, we will have a hope to restore order and peace on our streets in a place we call home.\nWe look forward to hearing from you soon.\nYour sincerely,Hong Kong Citizens\n(photo credit: Hong Kong Free Press)"],["topImageUrl","https://assets.change.org/photos/7/xd/mh/NmXdmHvvMnjVywm-800x450-noPad.jpg?1573414214"],["error",null]]}]}],["text","【請求荷蘭海牙常設仲裁法院調查香港暴警事件】\n這是具有法律效力的\n麻煩大家協助一下擴散跟連署！！\n\nhttps://www.change.org/p/request-permanent-court-of-arbitration-to-investigate-unlawful-use-of-force-of-hong-kong-police?utm_content=cl_sharecopy_17005953_en-GB%3Av5&recruiter=90670311&recruited_by_id=0040d850-5bec-43c4-8e81-c03a3953c79d&utm_source=share_petition&utm_medium=copylink&utm_campaign=psf_combo_share_message&utm_term=psf_combo_share_initial&share_bandit_exp=message-17005953-en-GB&share_bandit_var=v0"],["id","3mk40i15zdjrj"],["createdAt","2019-11-18T16:13:26.244Z"]]},{"__iterable":"Map","data":[["replyCount",1],["references",{"__iterable":"List","data":[{"__iterable":"Map","data":[["type","LINE"]]}]}],["replyRequestCount",1],["user",{"__iterable":"Map","data":[["id","j4S8C_8SU6w_lKU8142Pd-hyVJRULJNF7mjehuE3om7Vc6i8c"],["name","滿足的✖東勢✖艾薇拉"],["avatarUrl",null],["level",0]]}],["hyperlinks",{"__iterable":"List","data":[{"__iterable":"Map","data":[["title","Diabetes drug is promising Alzheimer's treatment after 'significantly reversing' memory loss"],["url","http://www.independent.co.uk/news/health/alzheimers-diabetes-drug-memory-loss-reverse-disease-treatment-degenerative-a8133681.html"],["summary","A drug developed for type 2 diabetes significantly reverses memory loss and could have potential as a new treatment for Alzheimer’s disease and other neurodegenerative diseases, scientists say.\n \n\n \n \n \n \n \n\n\n\n\n\n\n\n\n \n \n \n\n\n\n\n\n \n \n \n \n \n The study, by UK and Chinese universities, is the first to look at a new combined diabetes drug and found improvements in several characteristic symptoms of Alzheimer’s.\n \n\n \n \n \n \n \n\n\n\n \n\n\n\n\n\n\n\n \n \n \n\n\n\n\n\n \n \n \n \n \n Lead researcher Professor Christian Holscher, from Lancaster University, said these “very promising outcomes” show multi-action drugs developed for type 2 diabetes “consistently show neurological protective effects”.\n \n\n \n \n \n \n \n\n\n\n\n \n \n \n\n\n\n\n\n \n \n \n\n\n\n\n\n \n \n \n \n \n Independent academics said a reduction in nerve-cell-killing protein molecules was particularly interesting and this was likely to be another avenue in the search for an elusive drug to combat dementia.\n \n\n \n \n \n \n \n\n\n\n\n\n \n\n\n\n\n\n \n \n \n\n\n\n\n\n \n \n \n \n \n He has previously reported optimistic findings from an older diabetes drug, liraglutide, and clinical trials in humans are currently under way.\n \n\n \n \n \n \n \n\n\n\n\n\n\n\n\n \n \n \n\n\n\n\n\n \n \n \n \n \n This latest study, published in the journal Brain Research, looked at a “triple action” treatment that combine three different drugs for type 2 diabetes, acting on biological pathways that could also have an impact on dementia.\n \n\n \n \n \n \n \n\n\n\n\n\n\n\n\n \n \n \n \n \n\n\n\n\n\n \n \n \n \n \n \n \n\n \n \n \n Type 2 diabetes is a known risk factor for Alzhemier’s disease and impaired production of insulin – the hormone that people with diabetes don’t produce sufficiently to control their blood sugar – is linked to brain degeneration.\n \n\n \n \n \n \n \n\n\n\n\n\n\n\n\n \n \n \n\n\n\n\n\n \n \n \n \n \n The identification of this link had a twofold benefit, according to charities.\n \n\n \n \n \n \n \n\n\n\n\n\n\n\n\n \n \n \n\n\n\n\n\n \n \n \n \n \n It opened up new research and drug development opportunities, such as this study.\n \n\n \n \n \n \n \n\n\n\n\n\n\n\n\n \n \n \n\n\n\n\n\n \n \n \n \n \n But it also means that by making lifestyle changes, like eating healthily and exercising, patients can avoid developing type-2 diabetes and lower their risk of Alzheimer’s disease as well.\n \n\n \n \n \n \n \n\n\n\n\n\n\n\n\n \n \n \n\n\n\n\n\n \n \n \n \n \n There are currently half a million people living with Alzheimer’s disease in the UK and more than five million people with diabetes, both conditions pose a major cost to the NHS, and health systems worldwide.\n \n\n \n \n \n \n \n\n\n\n\n\n\n\n\n \n \n \n\n\n\n\n\n \n \n \n \n \n \n\n \n\n \n \n \n The treatment in this study combined; GLP-1, GIP and glucagon, which, like insulin protect against neurological deterioration, and tested them in mice with genetic mutations that cause Alzheimer’s disease.\n \n\n \n \n \n \n \n\n\n\n\n\n\n\n\n \n \n \n\n\n\n\n\n \n \n \n \n \n After two months of daily injections the mice were shown to significantly improve their performance in a maze designed to test memory.\n \n\n \n \n \n \n \n\n\n\n\n\n\n\n\n \n \n \n \n \n\n \n\n\n\n\n\n \n \n \n \n \n Treated mice also had lower levels of proteins which clump together and form plaques in the brains of people with Alzheimer’s disease, inhibiting nerve cells’ ability to communicate and causing them to die.\n \n\n \n \n \n \n \n\n\n\n\n\n\n\n\n \n \n \n\n\n\n\n\n \n \n \n \n \n The study also found these mice lost nerve cells to the disease at a slower rate and had lower levels of nerve inflammation.\n \n\n \n \n \n \n \n\n\n\n\n\n\n\n\n \n \n \n\n\n\n\n\n \n \n \n \n \n It concludes that the triple treatment “holds clear promise of being developed into a new treatment for chronic neurodegenerative disorders such as Alzheimer’s disease”.\n \n\n \n \n \n \n \n\n\n\n\n\n\n\n\n \n \n \n \n \n\n \n\n\n\n\n\n \n \n \n \n \n \n \nThe seven Alzheimer's risk factors\n\n\n\n\n \n\n\n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n\n\n \n\n \n \n \n The Alzheimer’s Society, a charity which part-funded the latest study research, said that after 15 years without a new drug for Alzheimer’s disease the promising results from these crossover treatments, which are already known to be safe for humans, are likely to bring earliest benefits.\n \n\n \n \n \n \n \n\n\n\n\n\n\n\n\n \n \n \n\n\n\n\n\n \n \n \n \n \n Professor Holscher added that further tests to compare its benefits against other potential treatments, and its effects in humans were needed.\n \n\n \n \n \n \n \n\n\n\n\n\n\n\n\n \n \n \n\n\n\n\n\n \n \n \n \n \n But Professor John Hardy, a professor of neuroscience, at University College London who was not involved in the trial, said the study was “a first step, at best” towards a Alzheimer’s drug for humans.\n \n\n \n \n \n \n \n\n\n\n\n\n\n\n\n \n \n \n \n \n\n \n\n\n\n\n\n \n \n \n \n \n “The results showing less amyloid deposition of amyloid in mice treated with glucagon receptor stimulating drugs is interesting,” he said.\n \n\n \n \n \n \n \n \n “However, it should be noted that several other drugs have shown positive results in mice models of Alzheimer’s disease and then failed in human trials.”\n \n\n \n \n \n \n \n \n Dr David Reynolds, chief scientific officer at Alzheimer’s Research UK, echoed this, but added: “There is a long road between studies that show an effect in animals and treatments in the hands of patients, and scientists will only be able to realise the potential of promising findings like these if we continue to invest in research.”"],["topImageUrl","https://static.independent.co.uk/s3fs-public/thumbnails/image/2015/04/14/21/web-alzheimers-2-alamy.jpg"],["error",null]]},{"__iterable":"Map","data":[["title","Neuroprotective effects of a triple GLP-1/GIP/glucagon receptor agonist in the APP/PS1 transgenic mouse model of Alzheimer's disease"],["url","https://www.sciencedirect.com/science/article/pii/S0006899317304596"],["summary","AbstractType 2 diabetes mellitus (T2DM) is a risk factor for Alzheimer disease (AD). Previous studies have shown that the incretin hormones glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic polypeptide (GIP) that have anti-diabetic properties show very promising effects in animal models of AD. Glucagon (Gcg) is a hormone and growth-factor, and the Gcg receptor is expressed in the brain. Here we test the effects of a triple receptor agonist (TA), which activates GIP-1, GIP and glucagon receptors at the same time. In the present study, the effects of the TA were evaluated in the APP/PS1 transgenic mouse model of AD. The TA was injected once-daily (10 nmol/kg i.p.) for two months. The results showed that treatment with TA significantly reversed the memory deficit in the APP/PS1 mice in a spatial water maze test. Moreover, the drug reduced levels of the mitochondrial pro-apoptotic signaling molecule BAX, increased the anti-apoptotic signaling molecule Bcl-2 and enhanced the levels of BDNF, a key growth factor that protects synaptic function. Levels of synaptophysin were enhanced, demonstrating protection from synaptic loss that is observed in AD. Neurogenesis in the dentate gyrus was furthermore enhanced as shown in the increase of doublecortin positive cells. Furthermore, TA treatment reduced the total amount of β-amyloid, reduced neuroinflammation (activated microglia and astrocytes), and oxidative stress in the cortex and hippocampus. Thus, these findings show that novel TAs are a promising lead for the design of future treatment strategies in AD.Previous Next KeywordsInflammationGrowth factorBDNFBrainInsulinNeurodegeneration1. IntroductionAlzheimer’s disease (AD), is a progressive neurodegenerative disease, characterized clinically by progressive memory loss, cognitive decline, and aberrant behavior [1]. Currently, there is no treatment can improve this condition. Epidemiological studies have shown that type 2 diabetes mellitus (T2DM) is a risk factor for Alzheimer disease (AD) (Luchsinger et al., 2004, Ohara et al., 2011). The underlying mechanism is most likely that insulin signaling is impaired in the brains of AD patients (Moloney et al., 2010, Talbot et al., 2012). This motivated research in drugs that have shown good effects in treating diabetes to investigate if they could be helpful in treating AD as well. Previous studies have shown that the incretin hormones glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic polypeptide (GIP) can play a neuroprotective role in the brain and show very promising effects in animal models of AD (Duffy and Holscher, 2013, Faivre and Holscher, 2013b, Li et al., 2010, McClean et al., 2011, McClean and Holscher, 2014). Glucagon (Gcg) is a hormone and growth-factor, and the Gcg receptor is expressed in the brain (Hoosein and Gurd, 1984, Mayo et al., 2003). Activation of the receptor can activate neuronal activity and cellular Ca2+ signaling and may have neuroprotective properties (Ayush et al., 2015). Here we test the effects of a triple receptor agonist (TA), which activates GIP-1, GIP and glucagon receptors at the same time. This novel drug has been developed as a potential treatment for diabetes (Finan et al., 2015). No studies have been published that test the neuroprotective effects of this novel drug. We therefore tested this promising TA in the APPSWE/PS1dE9 mouse model of AD. This mouse model recapitulates some of the hallmarks of AD, such as memory loss, synaptic loss, reduction of synaptic plasticity, reduction of neurogenesis in the dentate gyrus, chronic inflammation in the brain, and formation of amyloid plaques in the brain (Duffy and Holscher, 2013, Goto et al., 2008, Hamilton et al., 2011, Hamilton and Holscher, 2012, McClean et al., 2011). We tested the effects of TA in this transgenic mouse model and evaluated the neuroprotective effects. We analyzed memory formation, hippocampal neurogenesis, the expression levels of Brain Derived Neurotrophic Factor (BDNF) (Blurton-Jones et al., 2009), mitochondrial apoptosis signaling proteins, neuroinflammation and oxidative stress levels.2. Results2.1. TA-treatment improved the learning and memory impairment of APP/PS1 mice in the water mazeA two-way ANOVA found a significant difference between groups (p < 0.0001) and over time (p < 0.001). In the Morris water test, it was observed that the control mice, APP/PS1 mice and TA-treated APP/PS1 mice had a similar escape latency (P > 0.05). On day 5, the average escape distance in searching for the hidden platform in the APP/PS1 mice was significantly longer than that in the control mice (P < 0.05). Compared to the APP/PS1 group, the TA-treated APP/PS1 group’s average escape distance was decreased (P < 0.01). There was no difference in swimming speed among these three groups in training days (P > 0.05).Then on day 6, the platform was removed and the probe trail test was conducted. A one way ANOVA analysis showed an overall difference between groups. The APP/PS1 mice spent less time in the target quadrant (P < 0.01) and had fewer times passing through the original position of the hidden platform (P < 0.01), compared with control mice. In the TA-treated APP/PS1 group, the time spent in target quadrant (P < 0.05) and the frequencies to cross the platform significantly increased (P < 0.05), compared to the APP/PS1 group Fig. 1.Download high-res image (302KB)Download full-size imageFig. 1. The novel TA improved the learning ability of APP/PS1 mice in the spatial water maze task. (A) The escape latency and swimming speed in the training trails. (B) The percentage of time in target quadrant and crossing platform times in the probe trials. The circle indicates the position of the platform during acquisition (C) Representative swimming tracks of the probe trial. * = P < .05, ** = P < .01 compared to the control group, ## = P < .01, # = P < .05 compared to the APP/PS1 group; Two-way ANOVA with Bonferroni repeated measures post hoc tests.2.2. TA-treatment ameliorated amyloid plaques in the brains of APP/PS1 miceIn the immunohistochemical analysis, we analyzed the amount of amyloid plaques present in the brains of APP-PS1 mice. A one-way ANOVA found an overall difference on the amyloid deposits in the cortex (F = 31.10, P < 0.0001) and the hippocampus (F = 30.38, P < 0.0001) of mice between groups. The Aβ1-42 depositions in the cortex and hippocampus of the APP/PS1 mice were significantly higher than that in the control mice (P < 0.001), while TA-treatment group significantly decreased the deposition (P < 0.001) compared to the APP/PS1 group. N = 6 per group. See Fig. 2.Download high-res image (211KB)Download full-size imageFig. 2. TA treatment reduced amyloid deposits in the cortex and hippocampus of APP/PS1 mice. A, D: control group; B, E: APP/PS1 group; C, F: APP/PS1+TA group. Scale bar in image D: 100 μm. *** = P < .001 compared to the control group; ### = P < .001 compared to the APP/PS1 group. One-way ANOVA with Bonferroni repeated measures post hoc tests. N = 6 per group.2.3. TA-treatment reduced the chronic inflammation response in the cortex and hippocampus of the APP/PS1 miceIn the immunohistochemical analysis, an anti-GFAP antibody was used as an indication of activated astrocytes. A one-way ANOVA found an overall difference on the GFAP expression in the cortex (F = 77.03, P < 0.0001) and the hippocampus (F = 86.51, P < 0.0001) of mice between groups. The levels of GFAP in the cortex and hippocampus of the APP/PS1 mice were significantly higher than that in the control mice (P < 0.001), while TA-treatment group significantly decreased (P < 0.001) compared to the APP/PS1 group. N = 6 per group. See Fig. 3.Download high-res image (362KB)Download full-size imageFig. 3. TA-treatment reduced the number of GFAP positive cells in the cortex and hippocampus of APP/PS1 mice. A, D: control group; B, E: APP/PS1 group; C, F: APP/PS1+TA. Scale bar in image D: 50μm. A higher magnification image has been added to show stained astroglia. *** = P < .001 compared with the control group; ### = P < .001 compared with the APP/PS1 group. One-way ANOVA with Bonferroni repeated measures post hoc tests. N = 6 per group.In the immunohistochemical analysis, an anti-IBA1 antibody was used as an indication of microglia activation. A one-way ANOVA found an overall difference on the IBA-1 expression in the cortex (F = 28.71, P < 0.0001) and the hippocampus (F = 66.42, P < 0.0001) of mice between groups. The levels of IBA-1 in the cortex and hippocampus of the APP/PS1 mice were significantly higher than that in the control mice (P < 0.001), while TA-treatment group significantly decreased (P < 0.001, P < 0.01) compared to the APP/PS1 group. N = 6 per group. See Fig. 4.Download high-res image (347KB)Download full-size imageFig. 4. TA treatment reversed the increase of IBA-1 positive cells in the cortex and hippocampus of APP/PS1 mice. A, D: control group; B, E: APP/PS1 group; C, F: APP/PS1+TA group. Scale bar in image D: 100 μm. A higher magnification image has been added to show stained microglia. *** = P < .001 compared to the control group; ## = P < .01, ### = P < .001 compared to the APP/PS1 group. One-way ANOVA with Bonferroni repeated measures post hoc tests. N = 6 per group.2.4. TA-treatment suppressed oxidative stress in the cortex and hippocampus of APP /PS1 miceIn the immunohistochemical analysis, 4-HNE was measured as an indicator of lipid peroxidation. A one-way ANOVA found an overall difference on the 4-HNE expression in the cortex (F = 60.60, P < 0.0001) and the hippocampus (F = 35.22, P < 0.0001) of mice between groups. The expressions of 4-HNE in the cortex and hippocampus of the APP/PS1 mice were significantly higher than that in the control mice (P < 0.001), while TA-treatment group significantly decreased (P < 0.001) compared to the APP/PS1 group. N = 6 per group, see Fig. 5a.Download high-res image (397KB)Download full-size imageFig. 5. Oxidative stress was reduced by the TA. 5A. TA-treatment reduced the increase of 4-HNE levels in the cortex and hippocampus in APP/PS1 mice. A, D: control group; B, E: APP/PS1 group; C, F: APP/PS1+TA group. Scale bar in image D: 100 μm. *** = P < .001 compared with controls; ###=P < .001 compared to the APP/PS1 group. 5B. TA treatment reduced the increase of 8-OHdG levels in the cortex and hippocampus of APP/PS1 mice. A, D: control group; B, E: APP/PS1 group; C, F: APP/PS1+TA group. Scale bar in image D: 100 μm. *** = P < .001 compared to the control group; ## = P < .01, ### = P < .001 compared with the APP/PS1 group. One-way ANOVA with Bonferroni repeated measures ***post hoc tests. N = 6 per group.In the immunohistochemical analysis, 8-OHdG was measured as an indicator of DNA oxidation. A one-way ANOVN found an overall difference on the 8-OHdG expression in the cortex (F = 38.72, P < 0.0001) and the hippocampus (F = 56.60, P < 0.0001) of mice between groups. The expressions of 8-OHdG in the cortex and hippocampus of the APP/PS1 mice were significantly higher than that in the control mice (P < 0.001), while TA-treatment group significantly decreased (P < 0.01, P < 0.001) compared to the APP/PS1 group. N = 6 per group, see Fig. 5b.2.5. TA-treatment increased immature neurons in the dentate gyrus of APP/PS1 miceIn the immunohistochemical analysis, numbers of neurogenesis, assessed by doublecortin (DCX). A one-way ANOVN found an overall difference on the DCX positive cell in the dentate gyrus (F = 43.61, P < 0.0001) of mice between groups. The numbers of DCX were much reduced in the APP/PS1 mice (P < 0.001) compared to the control mice. While TA-treatment partially prevented the reduction (P < 0.001) compared to the APP/PS1 mice. N = 6 per group. See Fig. 6.Download high-res image (169KB)Download full-size imageFig. 6. TA-treatment enhanced neurogenesis and increased numbers of DCX positive cell numbers in the dentate gyrus of APP/PS1 mice. A: control group; B: APP/PS1 group; C: APP/PS1+TA group. Scale bar in image A: 100 μm. *** = P < .001 compared with the control group; ### = P < .001 compared with the APP/PS1 group. One-way ANOVA with Bonferroni repeated measures post hoc tests. N = 6 per group.2.6. TA-treatment reversed the decrease of BDNF levels in the hippocampus of APP/PS1 miceBDNF, a key neurotrophic factor in the brain, was detected by Western blot analysis. A one-way ANOVA found an overall difference between groups (F = 20.01, P < 0.001). The expression of BDNF in hippocampus was reduced in APP/PS1 group (P < 0.001) compared to the control group. TA-treatment reversed the decreased (P < 0.05) compared to the APP/PS1 group. N = 6 per group. See Fig. 7a.Download high-res image (304KB)Download full-size imageFig. 7. A) TA-treatment increased levels of BDNF in the hippocampus of APP/PS1 mice. *** = P < .001compared with the control group; # = P < .05 compared with the APP/PS1 group. N = 6 per group. B) TA-treatment increased levels of synaptophysin in the hippocampus of APP/PS1 mice. *** = P < .001 compared with the control group; ## = P < .01 compared with the APP/PS1 group. N = 6 per group. C) TA-treatment increased the ratio of BAX/Bcl-2 in the hippocampus of APP/PS1 mice. *** = P < .001 compared with the control group; ### = P < .001 compared with the APP/PS1 group. One-way ANOVA with Bonferroni repeated measures post hoc tests. N = 6 per group.2.7. TA-treatment reversed the reduction of synaptophysin levelsSynaptophysin (SYN), a synaptic markers, was detected by Western blot analysis. A one-way ANOVA found an overall difference between groups (F = 20.46, P < 0.001). The level of SYN in hippocampus was decreased in the APP/PS1 group (P < 0.001) compared to the control group. While TA-treatment prevent the reduction (P < 0.01) compared to the APP/PS1 group. N = 6 per group. See Fig. 7b.2.8. TA-treatment showed potential anti-apoptotic effects by decreasing the BAX/Bcl-2 ratio in APP/PS1 miceThe expression of apoptotic proteins, including Bcl-2 (that suppress apoptosis) and BAX (that promotes apoptosis), was detected by Western blot analysis. A one-way ANOVA found an overall difference between groups (F = 77.64, P < 0.001). The ratio of BAX/Bcl-2 in hippocampus was increased (P < 0.001) compared to the control group. While TA-treatment partly decreased the ratio of BAX/Bcl-2 (P < 0.001) compared to the APP/PS1 group. N = 6 per group. See Fig. 7c.3. DiscussionThe results demonstrate for the first time that the novel GLP-1/GIP/Gcg receptor agonist has clear neuroprotective effects in the APP/PS1 mouse model of AD. Memory formation in the spatial water maze task was improved by the drug, and the amyloid plaque load in the cortex and hippocampus was reduced. This result confirms our previous findings that single GLP-1 or GIP analogues can protect from memory loss in this mouse model of AD (Faivre and Holscher, 2013a, Faivre and Holscher, 2013b, McClean et al., 2011, McClean and Holscher, 2014). The chronic inflammation response as seen in the activation of microglia and astrocytes is much reduced by the novel TA, similar to single GLP-1 or GIP receptor agonists (Duffy and Holscher, 2013, Ji et al., 2016a, McClean et al., 2011). Levels of the peroxidized lipid 4-HNE that also acts as a pro-apoptotic signal in mitochondria have been reduced by the TA. 4-HNE is one of the markers of membrane lipid peroxidation induced by cytotoxic radicals such as OH. 4-HNE has cytotoxic, mutagenic and genotoxic properties and 4-HNE is reported to stimulate apoptosis via caspase 3 activation and by inducing mitochondrial damage as shown in cytochrome c release (Ji et al., 2001). The levels of the mitochondrial apoptosis signal BAX (Kim et al., 2005) were also reduced. Importantly, the anti-apoptotic signaling molecule B-cell lymphoma 2 (Bcl-2) which is active in functioning mitochondria that have normal inner membrane voltage potentials (Kavitha et al., 2013) is upregulated by the TA, adding to the neuroprotective effect. The level of chronic oxidative stress as measure by the levels of the oxidized nucleotide 8-Hydroxiguanosine (8-OHdG) was also reduced by the drug. Neurogenesis in the dentate gyrus was normalized. Importantly, the levels of the key growth factor BDNF were normalized in the brain. BDNF has shown clear neuroprotective effects in a range of animal models of disease (Allen et al., 2013, Benedetti et al., 2014, Blurton-Jones et al., 2009, Nagahara et al., 2013). An important factor of BNDF activity is that it maintains synaptic activity and protects synapses from stressors (Kuipers and Bramham, 2006). The observation that the synaptic marker synaptophysin was expressed at normal levels after TA treatment also indicates that synaptic numbers and function was improved in the AD mouse. We have shown previously that GLP-1 and GIP analogues protect synapses from amyloid induced stress and keeps them functional (Faivre and Holscher, 2013b, Gault and Holscher, 2008a, Gault and Holscher, 2008b, McClean and Holscher, 2014).Recently, multiple receptor agonists have been developed to treat type II diabetes (Finan et al., 2013, Finan et al., 2015). We have tested novel dual GLP-1/GIP receptor agonists that showed good neuroprotective effects that are superior to single GLP-1 analogues (Cao et al., 2016, Jalewa et al., 2017, Ji et al., 2016b, Yuan et al., 2017). The novel dual agonist DA3-CH was found to be superior to liraglutide in a mouse model of Parkinson’s disease (Yuan et al., 2017), demonstrating that the strategy of activating more than one incretin receptor type offers an advantage over single receptor agonists. Here, we show that a novel triple receptor agonist also shows promise as a potential treatment for AD, but further dose–response tests and direct comparisons with other drugs have to be conducted in order to evaluate if this new drug is superior to previous ones.GLP-1, GIP and glucagon are members of the growth factor family (Hölscher, 2016). Their respective receptors are classic 7-membrane spanning domain, G-protein coupled receptors that activate cAMP synthesis (Campbell and Drucker, 2013, Doyle and Egan, 2007, Holst et al., 2011). The activation of the second messenger cascade cAMP-PKA-CREB induces a number of cytoprotective processes, including the normalization of energy utilization, cell repair and growth-factor related gene expression (Holscher, 2014, Jalewa et al., 2016, Perry and Greig, 2004, Sharma et al., 2013). Importantly, GLP-1 and GIP also have anti-inflammatory properties. Astroglia and microglia express the receptors, and activating these reduce the inflammation response (Duffy and Holscher, 2013, Jalewa et al., 2017, Parthsarathy and Holscher, 2013, Spielman et al., 2017). Chronic inflammation is a main driver of neurodegenerative disorders, and a reduction of the inflammation in the brain shows protective effects (Akiyama et al., 2000, Clark et al., 2012, Lee et al., 2010).Based on the impressive preclinical data that demonstrate neuroprotection of such drugs, clinical trials have started that investigate the neuroprotective effects of the GLP-1 receptor agonists exendin-4 (Byetta, Bydureon®) and liraglutide (Victoza®) in patients with AD or with Parkinson disease (PD). A pilot study testing the GLP-1 analogue liraglutide in AD patients showed promising results. In a double blind, placebo controlled trial, AD patients showed no deterioration in brain activity as measured in the 18FDG-PET scan after 12 months of drug treatment. In contrast, the placebo group showed clear reductions in cortical activity as expected for this progressive neurodegenerative disease (Gejl et al., 2016). A phase II clinical trial is currently ongoing (Hölscher, 2016). A pilot trial testing exendin-4 in PD patients has shown good effects (NCT01174810). This open-label study tested the effects of exendin-4 in 45 patients. There were clear improvements in the motor coordination as shown in the MDS-UPDRS part 3 assessment, and cognition was improved as measured by the Mattis DRS-2 test battery that had been designed to evaluate cognitive impairments in PD patients (Aviles-Olmos et al., 2013). Patients were re-tested 12 months later, and the differences in motor performance and cognitive scores were still visible (Aviles-Olmos et al., 2014). A phase II double-blind, placebo controlled clinical trial that tested the once-weekly formulation of exendin-4, Bydureon®, confirmed the results of the pilot study. The drug group did not deteriorate in the MDS-UPDRS part 3 assessment after 48 weeks of treatment, and after 12 weeks of a wash-out period, the disease-modifying effect was still visible. In contrast, the placebo group deteriorated over time as would be expected in this disease (Athauda et al., 2017). In addition, a phase II trial testing liraglutide in PD patients has started, testing patients in a double blind, placebo controlled trial for one year (NCT02953665). A further clinical trial testing the GLP-1 receptor agonist Lixisenatide in PD patients is in planning.These very promising preclinical and the first clinical outcomes demonstrate the efficacy of these novel multiple receptor agonists that originally were developed to treat type II diabetes, but have shown consistent neuroprotective effects in several studies. In the present study, the novel TA improved learning and memory, reduce amyloid plaque load, chronic inflammation and oxidative stress, as well as mitochondrial stress signaling, and improved BDNF release and synaptic markers while normalising neurogenesis in the dentate gyrus. The novel triple GLP-1/GIP/Gcg receptor agonist holds clear promise of being developed into a new treatment for chronic neurodegenerative disorders such as Alzheimer’s disease.4. Materials and methods4.1. Peptide and chemicalsThe triple GLP-1/GIP/Gcg used in this study was synthesized by China Peptides Co, Ltd. (Shanghai, China) to 95% purity. The identity and purity of the peptide was confirmed by reversed-phase HPLC and characterized using matrix assisted laser resorption/ionization time of flight (MALDI–TOF) mass spectrometry. The peptide was stored in dry form and dissolved in double-distilled water containing 0.9% NaCl2 before experiments. A Bi-cinchoninicacid (BCA) protein assay kit was purchased from Apply gen Technologies Inc. (Beijing, China). 3, 3-diaminobenzidine (DAB) was purchased from ZSGB-BIO Co. (Beijing, China). The primary antibodies recognizing Aβ1-42, IBA-1, BDNF, 4-Hydroxynonenal (4-HNE), 8-Hydroxyguanosine (8-OHdG), synaptophysin (SYN) and β-actin were obtained from Abcam (Cambridge, UK). The primary antibody GFAP, Bcl-2, and BAX were obtained from Boster Biotechnology (Wuhan, China). The primary antibody doublecortin (DCX) buy from Santa Cruz Biotechnology. HRP-labeled goat anti-rabbit immunoglobulin was obtained from Abcam (Cambridge, UK). The biotinylated goat anti-rabbit IgG, biotinylated rabbit anti-goat IgG, avidinbiotin peroxidase complex reagent was purchased from Boster Biotechnology (Wuhan, China). All other reagents used in the research were of analytical grade and of the highest quality commercially available. The peptide sequence was as follows (Finan et al., 2015):HXQGTFTSDKSKYLDERAAQDFVQWLLDGGPSSGAPPPSX = aminoisobutyric acid, K = γE-C16 acylation (with a C16 fatty acid).4.2. Animals and treatmentsAPP/PS1 transgenic mice were purchased from the Academy of Military Medical Sciences (AMMS, China), were maintained in a constant temperature (22 ± 1 °C) and humidity (relative 30%) under a 12 h light/dark cycle in animal house. As well as age-matched non-AD mice, which are wild-type animals as control. When animals were 6 months of age, TA was dissolved in saline. The dose was a once-daily injection of 10 nmol/kg body weight ip. for two months. Injections were given in the morning (8:00 am). Control mice received equal volumes of saline solution. All experimental procedures involving animals were approved by the Institutional Animal Care Committee of Shanxi Medical University and conform to the guidelines of National Institute of Health (NIH) guideline (NIH publication No. 85-23. Revised 1985). They were divided into three different groups (n = 12). (i): control group treated with saline only; (ii): APP/PS1 group treated with saline only; (iii): APP/PS1 group treated with TA. At the end of drug treatments, measurements of behavioral change, neuronal damage, oxidative stress and inflammatory markers were conducted.4.3. Morris water maze testThe water maze test is a standard method for assessing learning and memory in animals. The water maze (WM) task was performed 5 days after two months intraperitoneal injection. The apparatus was a circular, white painted pool (150 cm in diameter and 60 cm in height) which was filled to a depth of 40 cm with warm water at a temperature 23 ± 2 °C to avoid hypothermia, and was surrounded by a white curtain around the pool. The pool was divided into four quadrants with four starting locations, north (N), east (E), south (S) and west (W), the starting quadrant was randomly chosen. A perspex platform (diameter, 10 cm) was located in a specific location during the acquisition trials and submerged approximately 1.0 cm below the surface of the water. A video camera was mounted directly above the pool to record the movement and the trail of the mice. During the acquisition phase, each mouse was subjected to four consecutive training trials on each day. The mouse was gently placed in the water between the quadrants, and allowed 60 s to search the hidden platform. If it failed to find the platform within 60 s, it was guided to the platform and allowed to remain there for 10 s. Training continued for 5 days. On the sixth day, the hidden platform was removed and each mouse was allowed to swim in the pool for 60 s. The mean time spent in all four quadrants and the target area for searching the hidden platform was recorded.4.4. Brain tissue preparationHalf of the animals were decapitated after anesthetization with ethyl carbamate, hippocampus tissues were immediately isolated and stored at −80 °C for further investigations. The second half of each group was used for immunohistochemistry analysis. Animals were perfused intracardially with saline and cold 4% paraformaldehyde. Brains were removed and post-fixed in 4% paraformaldehyde for 24 h.4.5. ImmunohistochemistryFor DAB staining, fixed tissues were embedded in paraffin. Coronal sections (5 μm thick) of the brains were cut on a Leica microtome for immunohistochemistry. Paraffin was removed from the tissue sections with xylene, and the sections were rehydrated in graded ethanol solutions. Endogenous peroxidase activity was blocked with 3% H2O2 for 10 min at room temperature. Antigen retrieval was performed by heating in 10 mmol/L citrate buffer (pH 6.0) for 10 min. After blocked with 5% BSA, sections were incubated with the primary antibody for doublecortin (goat anti-DCX; 1:50), GFAP (rabbit anti-GFAP; 1:100), IBA-1 (goat anti-IBA1; 1:200), 4-Hydroxynonenal antibody (goat anti-4 HNE; 1:200) and anti-8 Hydroxyguanosine antibody (goat anti-8 OHdG; 1:200) at 37 °C for 2 h. For the Anti-8 OHdG histology, enzymatic antigen retrieval with 10 μm/ml proteinase K for 40 min at 37 °C was performed. To detect amyloid deposits, the primary antibody was Aβ1-42 (rabbit anti-Aβ1-42; 1:100), brain sections were pre-incubated with 70% formic acid and incubated at 4 °C for 24 h. They were rinsed in PBS and incubated with biotinylated goat anti-rabbit IgG or biotinylated rabbit anti-goat IgG at 37 °C for 0.5 h and incubated with the avidin-biotin peroxidase complex reagent at 37 °C for 0.5 h. Samples were stained in DAB solution in the presence of H2O2. All stained sections were viewed and photographed under a Zeiss light microscope, and images were captured by Motic Images Plus2.0ML software. The numbers of DCX positive neurons, the areas of Aβ1-42, GFAP, IBA-1, 4-HNE and 8-OHdG positive cells were determined using Image-pro plus6.0 software. The software counts objects that are defined in size and staining threshold by the user. This automated process avoided user bias.4.6. Western blotThe hippocampal tissues from mice was homogenized in ice cold RIPA buffer (containing 1% Triton X-100, 0.1% SDS, 1% deoxycholate) and PMSF (phenyl-methylsulfonyl fluoride). The homogenate was centrifuged 12,000 rpm/min for 5 min at 4 °C before taking the supernatant. Tissue containing 60 μg of protein was separated on 10–12% gradient Bis–Tris gel with molecular marker and electrophoresed in running buffer at 120 mV for 90 min followed by transfer to the PVDF membrane. Following protein transfer, the membrane was washed in TBST (tris buffered saline with 0.05% Tween-20, pH 8) and blocked in 5% skim milk for 1 h. Then incubated overnight at 4 °C with rabbit anti-synaptophysin (SYN) (1:10,000), rabbit anti-Bcl-2 (1:400), rabbit anti-BAX (1:400), rabbit anti-BDNF (1:5000), after three washes in TBS further incubation for 2 h at room temperature with the HRP-labeled goat anti-rabbit immunoglobulin (1:5000). In some cases, the blots were stripped and reprobed with a rabbit anti-β-actin (1:5000) to ensure equal sample loading. The bound antibodies were then visualized by ECL-enhanced chemiluminescence (Beyotime Institute of Biotechnology, Shanghai, China) according to the manufacturer's instructions. Western blot images were captured with a chemiluminescent imaging system (Sage Creation, Beijing, China). All bands were quantified using the image system of Alpha View SA.4.7. Statistical analysisThe data were presented as the mean ± standard error of mean (S.E.M.) and analyzed by one-way or two way ANOVA followed by Bonferroni’s multiple comparison test using the software Prism 5 (Graph Pad Software Inc, La Jolla, CA, USA). Statistical significance is defined as P < 0.05.ContributorsJT, WL and YL conducted the experiments, LL and CH conceived the experiment and wrote the manuscript.AcknowledgementsThis work was funded by the Ministry of Human Resources and Social Security, Shanxi Province [(2010)255], by a Shanxi Scholarship Council of China, and supported by a grant from the Alzheimer Society UK. The authors declare no conflict of interestAppendix A. Supplementary dataReferencesAkiyama et al., 2000H. Akiyama, S. Barger, S. Barnum, B. Bradt, J. Bauer, G.M. Cole, N.R. Cooper, P. Eikelenboom, M. Emmerling, B.L. Fiebich, C.E. Finch, S. Frautschy, W.S. Griffin, H. Hampel, M. Hull, G. Landreth, L. Lue, R. Mrak, I.R. Mackenzie, P.L. McGeer, M.K. O'Banion, J. Pachter, G. Pasinetti, C. Plata-Salaman, J. Rogers, R. Rydel, Y. Shen, W. Streit, R. Strohmeyer, I. Tooyoma, F.L. Van Muiswinkel, R. Veerhuis, D. Walker, S. Webster, B. Wegrzyniak, G. Wenk, T. Wyss-CorayInflammation and Alzheimer’s diseaseNeurobiol. Aging, 21 (2000), pp. 383-421Allen et al., 2013S.J. Allen, J.J. Watson, D.K. Shoemark, N.U. Barua, N.K. PatelGDNF, NGF and BDNF as therapeutic options for neurodegenerationPharmacol. Ther., 138 (2013), pp. 155-175Athauda et al., 2017D. Athauda, K. Maclagan, S.S. Skene, M. Bajwa-Joseph, D. Letchford, K. Chowdhury, S. Hibbert, N. Budnik, L. Zampedri, J. Dickson, Y. Li, I. Aviles-Olmos, T.T. Warner, P. Limousin, A.J. Lees, N.H. Greig, S. Tebbs, T. FoltynieExenatide once weekly versus placebo in Parkinson’s disease: a randomised, double-blind, placebo-controlled trialAviles-Olmos et al., 2013I. Aviles-Olmos, J. Dickson, Z. Kefalopoulou, A. Djamshidian, P. Ell, T. Soderlund, P. Whitton, R. Wyse, T. Isaacs, A. Lees, P. Limousin, T. FoltynieExenatide and the treatment of patients with Parkinson’s diseaseJ. Clin. Invest., 123 (2013), pp. 2730-2736Aviles-Olmos et al., 2014I. Aviles-Olmos, J. Dickson, Z. Kefalopoulou, A. Djamshidian, J. Kahan, P.E. Fmedsci, P. Whitton, R. Wyse, T. Isaacs, A. Lees, P. Limousin, T. FoltynieMotor and cognitive advantages persist 12 months after exenatide exposure in Parkinson’s diseaseJ. Parkinson’s Dis., 4 (2014), pp. 337-344Ayush et al., 2015E.A. Ayush, Y. Iwasaki, S. Iwamoto, H. Nakabayashi, M. Kakei, T. YadaGlucagon directly interacts with vagal afferent nodose ganglion neurons to induce Ca(2+) signaling via glucagon receptorsBiochem. Biophys. Res. Commun., 456 (2015), pp. 727-732Benedetti et al., 2014E. Benedetti, B. D'Angelo, L. Cristiano, E. Di Giacomo, F. Fanelli, S. Moreno, F. Cecconi, A. Fidoamore, A. Antonosante, R. Falcone, R. Ippoliti, A. Giordano, A. CiminiInvolvement of peroxisome proliferator-activated receptor beta/delta (PPAR beta/delta) in BDNF signaling during aging and in Alzheimer disease: possible role of 4-hydroxynonenal (4-HNE)Cell Cycle, 13 (2014), pp. 1335-1344Blurton-Jones et al., 2009M. Blurton-Jones, M. Kitazawa, H. Martinez-Coria, N.A. Castello, F.J. Muller, J.F. Loring, T.R. Yamasaki, W.W. Poon, K.N. Green, F.M. LaferlaNeural stem cells improve cognition via BDNF in a transgenic model of Alzheimer diseaseProc. Natl. Acad. Sci. U.S.A., 106 (2009), pp. 13594-13599Campbell and Drucker, 2013J.E. Campbell, D.J. DruckerPharmacology, physiology, and mechanisms of incretin hormone actionCell Metab., 17 (2013), pp. 819-837Cao et al., 2016L. Cao, D. Li, P. Feng, L. Li, G. Xue, G. Li, C. HölscherA novel dual GLP-1 and GIP incretin receptor agonist is neuroprotective in a mouse model of Parkinson’s disease by reducing chronic inflammation in the brainNeuroreport, 37 (2016), pp. 384-391Clark et al., 2012I. Clark, C. Atwood, R. Bowen, G. Paz-Filho, B. VisselTumor necrosis factor-induced cerebral insulin resistance in Alzheimer's disease links numerous treatment rationalesPharmacol. Rev., 64 (2012), pp. 1004-1026Doyle and Egan, 2007M.E. Doyle, J.M. EganMechanisms of action of glucagon-like peptide 1 in the pancreasPharmacol. Ther., 113 (2007), pp. 546-593Duffy and Holscher, 2013A.M. Duffy, C. HolscherThe incretin analogue D-Ala(2)GIP reduces plaque load, astrogliosis and oxidative stress in an APP/PS1 mouse model of Alzheimer’s diseaseNeuroscience, 228 (2013), pp. 294-300Faivre and Holscher, 2013aE. Faivre, C. HolscherD-Ala2GIP facilitated synaptic plasticity and reduces plaque load in aged wild type mice and in an Alzheimer's disease mouse modelJ. Alzheimers Dis., 35 (2013), pp. 267-283Faivre and Holscher, 2013bE. Faivre, C. HolscherNeuroprotective effects of D-Ala2GIP on Alzheimer’s disease biomarkers in an APP/PS1 mouse modelAlzheimers Res. Ther., 5 (2013), pp. 20-28Finan et al., 2013B. Finan, T. Ma, N. Ottaway, T.D. Muller, K.M. Habegger, K.M. Heppner, H. Kirchner, J. Holland, J. Hembree, C. Raver, S.H. Lockie, D.L. Smiley, V. Gelfanov, B. Yang, S. Hofmann, D. Bruemmer, D.J. Drucker, P.T. Pfluger, D. Perez-Tilve, J. Gidda, L. Vignati, L. Zhang, J.B. Hauptman, M. Lau, M. Brecheisen, S. Uhles, W. Riboulet, E. Hainaut, E. Sebokova, K. Conde-Knape, A. Konkar, R.D. DiMarchi, M.H. TschopUnimolecular dual incretins maximize metabolic benefits in rodents, monkeys, and humansSci. Transl. Med., 5 (2013), p. 209ra151Finan et al., 2015B. Finan, B. Yang, N. Ottaway, D.L. Smiley, T. Ma, C. Clemmensen, J. Chabenne, L. Zhang, K.M. Habegger, K. Fischer, J.E. Campbell, D. Sandoval, R.J. Seeley, K. Bleicher, S. Uhles, W. Riboulet, J. Funk, C. Hertel, S. Belli, E. Sebokova, K. Conde-Knape, A. Konkar, D.J. Drucker, V. Gelfanov, P.T. Pfluger, T.D. Muller, D. Perez-Tilve, R.D. DiMarchi, M.H. TschopA rationally designed monomeric peptide triagonist corrects obesity and diabetes in rodentsNat. Med., 21 (2015), pp. 27-36Gault and Holscher, 2008aV.A. Gault, C. HolscherGLP-1 agonists facilitate hippocampal LTP and reverse the impairment of LTP induced by beta-amyloidEur. J. Pharmacol., 587 (2008), pp. 112-117Gault and Holscher, 2008bV.A. Gault, C. HolscherProtease-resistant glucose-dependent insulinotropic polypeptide agonists facilitate hippocampal LTP and reverse the impairment of LTP induced by beta-amyloidJ. Neurophysiol., 99 (2008), pp. 1590-1595Gejl et al., 2016M. Gejl, A. Gjedde, L. Egefjord, A. Møller, S.B. Hansen, K. Vang, A.B. Rodell, H. Braendgaard, H. Gottrup, A. Schacht, N. Møller, B. Brock, J. RungbyIn Alzheimer’s disease, six-month treatment with glp-1 analogue prevents decline of brain glucose metabolism: randomized, placebo-controlled double-blind clinical trialFront. Aging Neurosci., 8 (2016), pp. 1-10Goto et al., 2008Y. Goto, T. Niidome, H. Hongo, A. Akaike, T. Kihara, H. SugimotoImpaired muscarinic regulation of excitatory synaptic transmission in the APPswe/PS1dE9 mouse model of Alzheimer's diseaseEur. J. Pharmacol., 583 (2008), pp. 84-91Hamilton et al., 2011A. Hamilton, S. Patterson, D. Porter, V.A. Gault, C. HolscherNovel GLP-1 mimetics developed to treat type 2 diabetes promote progenitor cell proliferation in the brainJ. Neurosci. Res., 89 (2011), pp. 481-489Hamilton and Holscher, 2012A. Hamilton, C. HolscherThe effect of ageing on neurogenesis and oxidative stress in the APP(swe)/PS1(deltaE9) mouse model of Alzheimer's diseaseBrain Res., 1449 (2012), pp. 83-93Holscher, 2014C. HolscherInsulin, incretins and other growth factors as potential novel treatments for Alzheimer's and Parkinson's diseasesBiochem. Soc. Trans., 42 (2014), pp. 593-599Hölscher, 2016C. HölscherGLP-1 and GIP analogues as novel treatments for Alzheimer’s and Parkinson’s diseaseCardiovasc. Endocrinol., 5 (2016), pp. 93-98Holst et al., 2011J.J. Holst, R. Burcelin, E. NathansonNeuroprotective properties of GLP-1: theoretical and practical applicationsCurr. Med. Res. Opin., 27 (2011), pp. 547-558Hoosein and Gurd, 1984N.M. Hoosein, R.S. GurdIdentification of glucagon receptors in rat brainProc. Natl. Acad. Sci. U.S.A., 81 (1984), pp. 4368-4372Jalewa et al., 2016J. Jalewa, M.K. Sharma, C. HolscherNovel incretin analogues improve autophagy and protect from mitochondrial stress induced by rotenone in SH-SY5Y cellsJ. Neurochem., 139 (2016), pp. 55-67Jalewa et al., 2017J. Jalewa, M. Sharma, S. Gengler, C. HölscherA novel GLP-1/GIP dual receptor agonist protects from 6-OHDA lesion in a rat model of Parkinson’s diseaseNeuropharmacology, 117 (2017), pp. 238-248Ji et al., 2001C. Ji, V. Amarnath, J.A. Pietenpol, L.J. Marnett4-hydroxynonenal induces apoptosis via caspase-3 activation and cytochrome c releaseChem. Res. Toxicol., 14 (2001), pp. 1090-1096Ji et al., 2016aC. Ji, G.F. Xue, G. Li, D. Li, C. HolscherNeuroprotective effects of glucose-dependent insulinotropic polypeptide in Alzheimer's diseaseRev. Neurosci., 27 (2016), pp. 61-70Ji et al., 2016bC. Ji, G.F. Xue, C. Lijun, P. Feng, D. Li, L. Li, G. Li, C. HolscherA novel dual GLP-1 and GIP receptor agonist is neuroprotective in the MPTP mouse model of Parkinson's disease by increasing expression of BNDFBrain Res., 1634 (2016), pp. 1-11Kavitha et al., 2013M. Kavitha, J. Nataraj, M.M. Essa, M.A. Memon, T. ManivasagamMangiferin attenuates MPTP induced dopaminergic neurodegeneration and improves motor impairment, redox balance and Bcl-2/Bax expression in experimental Parkinson's disease miceChem. Biol. Interact., 206 (2013), pp. 239-247Kim et al., 2005S.J. Kim, K. Winter, C. Nian, M. Tsuneoka, Y. Koda, C.H. McIntoshGlucose-dependent insulinotropic polypeptide (GIP) stimulation of pancreatic beta-cell survival is dependent upon phosphatidylinositol 3-kinase (PI3K)/protein kinase B (PKB) signaling, inactivation of the forkhead transcription factor Foxo1, and down-regulation of bax expressionJ. Biol. Chem., 280 (2005), pp. 22297-22307Kuipers and Bramham, 2006S.D. Kuipers, C.R. BramhamBrain-derived neurotrophic factor mechanisms and function in adult synaptic plasticity: new insights and implications for therapyCurr. Opin. Drug Discov. Dev., 9 (2006), pp. 580-586Lee et al., 2010Y.J. Lee, S.B. Han, S.Y. Nam, K.W. Oh, J.T. HongInflammation and Alzheimer's diseaseArch. Pharm. Res., 33 (2010), pp. 1539-1556Li et al., 2010Y. Li, K. Duffy, M. Ottinger, B. Ray, J. Bailey, H. Holloway, D. Tweedie, T. Perry, M. Mattson, D. Kapogiannis, K. Sambamurti, D. Lahiri, N. GreigGLP-1 receptor stimulation reduces amyloid-beta peptide accumulation and cytotoxicity in cellular and animal models of Alzheimer's diseaseJ. Alzheimers Dis., 19 (2010), pp. 1205-1219Luchsinger et al., 2004J.A. Luchsinger, M.X. Tang, S. Shea, R. MayeuxHyperinsulinemia and risk of Alzheimer diseaseNeurology, 63 (2004), pp. 1187-1192Mayo et al., 2003K.E. Mayo, L.J. Miller, D. Bataille, S. Dalle, B. Goke, B. Thorens, D.J. DruckerInternational Union of Pharmacology. XXXV. The glucagon receptor familyPharmacol. Rev., 55 (2003), pp. 167-194McClean et al., 2011P. McClean, V. Parthsarathy, E. Faivre, C. HölscherThe diabetes drug liraglutide prevents degenerative processes in a mouse model of Alzheimer's diseaseJ. Neurosci., 31 (2011), pp. 6587-6594McClean and Holscher, 2014P.L. McClean, C. HolscherLixisenatide, a drug developed to treat type 2 diabetes, shows neuroprotective effects in a mouse model of Alzheimer's diseaseNeuropharmacology, 86C (2014), pp. 241-258Moloney et al., 2010A.M. Moloney, R.J. Griffin, S. Timmons, R. O'Connor, R. Ravid, C. O'NeillDefects in IGF-1 receptor, insulin receptor and IRS-1/2 in Alzheimer's disease indicate possible resistance to IGF-1 and insulin signallingNeurobiol. Aging, 31 (2010), pp. 224-243Nagahara et al., 2013A.H. Nagahara, M. Mateling, I. Kovacs, L. Wang, S. Eggert, E. Rockenstein, E.H. Koo, E. Masliah, M.H. TuszynskiEarly BDNF treatment ameliorates cell loss in the entorhinal cortex of APP transgenic miceJ. Neurosci., 33 (2013), pp. 15596-15602Ohara et al., 2011T. Ohara, Y. Doi, T. Ninomiya, Y. Hirakawa, J. Hata, T. Iwaki, S. Kanba, Y. KiyoharaGlucose tolerance status and risk of dementia in the community: the Hisayama studyNeurology, 77 (2011), pp. 1126-1134Parthsarathy and Holscher, 2013V. Parthsarathy, C. HolscherThe type 2 diabetes drug liraglutide reduces chronic inflammation induced by irradiation in the mouse brainEur. J. Pharmacol., 700 (2013), pp. 42-50Perry and Greig, 2004T.A. Perry, N.H. GreigA new Alzheimer's disease interventive strategy: GLP-1Curr. Drug Targets, 5 (2004), pp. 565-571Sharma et al., 2013M. Sharma, J. Jalewa, C. HolscherNeuroprotective and anti-apoptotic effects of liraglutide on SH-SY5Y cells exposed to Methylglyoxal stressJ. Neurochem., 128 (2013), pp. 459-471Spielman et al., 2017L.J. Spielman, D.L. Gibson, A. KlegerisIncretin hormones regulate microglia oxidative stress, survival and expression of trophic factorsEur. J. Cell Biol., 96 (2017), pp. 240-253Talbot et al., 2012K. Talbot, H.Y. Wang, H. Kazi, L.Y. Han, K.P. Bakshi, A. Stucky, R.L. Fuino, K.R. Kawaguchi, A.J. Samoyedny, R.S. Wilson, Z. Arvanitakis, J.A. Schneider, B.A. Wolf, D.A. Bennett, J.Q. Trojanowski, S.E. ArnoldDemonstrated brain insulin resistance in Alzheimer's disease patients is associated with IGF-1 resistance, IRS-1 dysregulation, and cognitive declineJ. Clin. Invest., 122 (2012), pp. 1316-1338Yuan et al., 2017Z. Yuan, L. Li, P. Feng, G. Xue, C. Ji, G. Li, C. HölscherA novel GLP-1/GIP dual agonist is more effective than liraglutide in reducing inflammation and enhancing GDNF release in the MPTP mouse model of Parkinson’s disease© 2017 Elsevier B.V. All rights reserved."],["topImageUrl","https://ars.els-cdn.com/content/image/1-s2.0-S0006899317X0022X-cov150h.gif"],["error",null]]}]}],["text","阿茲海默症(Alzheimer’s disease)疾病對於病人及家屬皆會帶來很大的影響，然而至目前為止，疾病的發生機轉及可能的治療方式卻仍在研究階段。《Cell Reports》日前刊登中國山西醫科大學(Shanxi Medical University)的最新發表，研究團隊表示，原先發展用來治療糖尿病的特定藥物，有機會反轉阿茲海默症的記憶喪失症狀。第二型糖尿病已知為阿茲海默症的風險因子之一，其病程會影響到胰島素的分泌，而在過去的研究中亦曾發現，這個影響人體血糖調節的荷爾蒙，其分泌不足可能與大腦的退化情形有關。本篇研究中，作者發現結合第二型糖尿病的特定藥物，可能改善數項阿茲海默症的獨特病徵。作者表示，這項研究發現，有助於未來發展阿茲海默症及其它神經退化疾病的新治療方法。\n \n新聞出處：Alzheimer's treatment: Diabetes drug holds promise for fighting disease after 'significantly reversing' memory loss《Independent》\nhttp://www.independent.co.uk/news/health/alzheimers-diabetes-drug-memory-loss-reverse-disease-treatment-degenerative-a8133681.html\n研究刊登在《Cell Reports》第1678卷第64-74頁\nhttps://www.sciencedirect.com/science/article/pii/S0006899317304596\n "],["id","AWEET-BFyCdS-nWhunGr"],["createdAt","2018-01-17T13:29:59.107Z"]]},{"__iterable":"Map","data":[["replyCount",1],["references",{"__iterable":"List","data":[{"__iterable":"Map","data":[["type","LINE"]]}]}],["replyRequestCount",2],["user",{"__iterable":"Map","data":[["id","j4S8C_YlgNaHHUAmuAJvBJ9fT06nPwQIphzAJEbrZaWWhlh_8"],["name","發起者的頭屋維拉"],["avatarUrl",null],["level",0]]}],["hyperlinks",{"__iterable":"List","data":[{"__iterable":"Map","data":[["title","Respiratory virus shedding in exhaled breath and efficacy of face masks"],["url","https://www.nature.com/articles/s41591-020-0843-2?fbclid=IwAR3EzP6Xm2bXH4bDHx9i9oOYxqHIraWbuRbZcGzM6fmgQz7iIl7ufHrkN-A"],["summary","AbstractWe identified seasonal human coronaviruses, influenza viruses and rhinoviruses in exhaled breath and coughs of children and adults with acute respiratory illness. Surgical face masks significantly reduced detection of influenza virus RNA in respiratory droplets and coronavirus RNA in aerosols, with a trend toward reduced detection of coronavirus RNA in respiratory droplets. Our results indicate that surgical face masks could prevent transmission of human coronaviruses and influenza viruses from symptomatic individuals.\n \n \n \n\n \n \n MainRespiratory virus infections cause a broad and overlapping spectrum of symptoms collectively referred to as acute respiratory virus illnesses (ARIs) or more commonly the ‘common cold’. Although mostly mild, these ARIs can sometimes cause severe disease and death1. These viruses spread between humans through direct or indirect contact, respiratory droplets (including larger droplets that fall rapidly near the source as well as coarse aerosols with aerodynamic diameter >5 µm) and fine-particle aerosols (droplets and droplet nuclei with aerodynamic diameter ≤5 µm)2,3. Although hand hygiene and use of face masks, primarily targeting contact and respiratory droplet transmission, have been suggested as important mitigation strategies against influenza virus transmission4, little is known about the relative importance of these modes in the transmission of other common respiratory viruses2,3,5. Uncertainties similarly apply to the modes of transmission of COVID-19 (refs. 6,7).Some health authorities recommend that masks be worn by ill individuals to prevent onward transmission (source control)4,8. Surgical face masks were originally introduced to protect patients from wound infection and contamination from surgeons (the wearer) during surgical procedures, and were later adopted to protect healthcare workers against acquiring infection from their patients. However, most of the existing evidence on the filtering efficacy of face masks and respirators comes from in vitro experiments with nonbiological particles9,10, which may not be generalizable to infectious respiratory virus droplets. There is little information on the efficacy of face masks in filtering respiratory viruses and reducing viral release from an individual with respiratory infections8, and most research has focused on influenza11,12.Here we aimed to explore the importance of respiratory droplet and aerosol routes of transmission with a particular focus on coronaviruses, influenza viruses and rhinoviruses, by quantifying the amount of respiratory virus in exhaled breath of participants with medically attended ARIs and determining the potential efficacy of surgical face masks to prevent respiratory virus transmission.ResultsWe screened 3,363 individuals in two study phases, ultimately enrolling 246 individuals who provided exhaled breath samples (Extended Data Fig. 1). Among these 246 participants, 122 (50%) participants were randomized to not wearing a face mask during the first exhaled breath collection and 124 (50%) participants were randomized to wearing a face mask. Overall, 49 (20%) voluntarily provided a second exhaled breath collection of the alternate type.Infections by at least one respiratory virus were confirmed by reverse transcription PCR (RT–PCR) in 123 of 246 (50%) participants. Of these 123 participants, 111 (90%) were infected by human (seasonal) coronavirus (n = 17), influenza virus (n = 43) or rhinovirus (n = 54) (Extended Data Figs. 1 and 2), including one participant co-infected by both coronavirus and influenza virus and another two participants co-infected by both rhinovirus and influenza virus. These 111 participants were the focus of our analyses.There were some minor differences in characteristics of the 111 participants with the different viruses (Table 1a). Overall, 24% of participants had a measured fever ≥37.8 °C, with patients with influenza more than twice as likely than patients infected with coronavirus and rhinovirus to have a measured fever. Coronavirus-infected participants coughed the most with an average of 17 (s.d. = 30) coughs during the 30-min exhaled breath collection. The profiles of the participants randomized to with-mask versus without-mask groups were similar (Supplementary Table 1).Table 1a Characteristics of individuals with symptomatic coronavirus, influenza virus or rhinovirus infectionWe tested viral shedding (in terms of viral copies per sample) in nasal swabs, throat swabs, respiratory droplet samples and aerosol samples and compared the latter two between samples collected with or without a face mask (Fig. 1). On average, viral shedding was higher in nasal swabs than in throat swabs for each of coronavirus (median 8.1 log10 virus copies per sample versus 3.9), influenza virus (6.7 versus 4.0) and rhinovirus (6.8 versus 3.3), respectively. Viral RNA was identified from respiratory droplets and aerosols for all three viruses, including 30%, 26% and 28% of respiratory droplets and 40%, 35% and 56% of aerosols collected while not wearing a face mask, from coronavirus, influenza virus and rhinovirus-infected participants, respectively (Table 1b). In particular for coronavirus, we identified OC43 and HKU1 from both respiratory droplets and aerosols, but only identified NL63 from aerosols and not from respiratory droplets (Supplementary Table 2 and Extended Data Fig. 3).Fig. 1: Efficacy of surgical face masks in reducing respiratory virus shedding in respiratory droplets and aerosols of symptomatic individuals with coronavirus, influenza virus or rhinovirus infection.a–c, Virus copies per sample collected in nasal swab (red), throat swab (blue) and respiratory droplets collected for 30min while not wearing (dark green) or wearing (light green) a surgical face mask, and aerosols collected for 30min while not wearing (brown) or wearing (orange) a face mask, collected from individuals with acute respiratory symptoms who were positive for coronavirus (a), influenza virus (b) and rhinovirus (c), as determined by RT–PCR in any samples. P values for mask intervention as predictor of log10 virus copies per sample in an unadjusted univariate Tobit regression model which allowed for censoring at the lower limit of detection of the RT–PCR assay are shown, with significant differences in bold. For nasal swabs and throat swabs, all infected individuals were included (coronavirus, n=17; influenza virus, n=43; rhinovirus, n=54). For respiratory droplets and aerosols, numbers of infected individuals who provided exhaled breath samples while not wearing or wearing a surgical face mask, respectively were: coronavirus (n=10 and 11), influenza virus (n=23 and 28) and rhinovirus (n=36 and 32). A subset of participants provided exhaled breath samples for both mask interventions (coronavirus, n=4; influenza virus, n=8; rhinovirus, n=14). The box plots indicate the median with the interquartile range (lower and upper hinge) and ±1.5×interquartile range from the first and third quartile (lower and upper whiskers).Table 1b Efficacy of surgical face masks in reducing respiratory virus frequency of detection and viral shedding in respiratory droplets and aerosols of symptomatic individuals with coronavirus, influenza virus or rhinovirus infectionWe detected coronavirus in respiratory droplets and aerosols in 3 of 10 (30%) and 4 of 10 (40%) of the samples collected without face masks, respectively, but did not detect any virus in respiratory droplets or aerosols collected from participants wearing face masks, this difference was significant in aerosols and showed a trend toward reduced detection in respiratory droplets (Table 1b). For influenza virus, we detected virus in 6 of 23 (26%) and 8 of 23 (35%) of the respiratory droplet and aerosol samples collected without face masks, respectively. There was a significant reduction by wearing face masks to 1 of 27 (4%) in detection of influenza virus in respiratory droplets, but no significant reduction in detection in aerosols (Table 1b). Moreover, among the eight participants who had influenza virus detected by RT–PCR from without-mask aerosols, five were tested by viral culture and four were culture-positive. Among the six participants who had influenza virus detected by RT–PCR from with-mask aerosols, four were tested by viral culture and two were culture-positive. For rhinovirus, there were no significant differences between detection of virus with or without face masks, both in respiratory droplets and in aerosols (Table 1b). Conclusions were similar in comparisons of viral shedding (Table 1b). In addition, we found a significant reduction in viral shedding (Supplementary Table 2) in respiratory droplets for OC43 (Extended Data Fig. 4) and influenza B virus (Extended Data Fig. 5) and in aerosols for NL63 (Extended Data Fig. 4).We identified correlations between viral loads in different samples (Extended Data Figs. 6–8) and some evidence of declines in viral shedding by time since onset for influenza virus but not for coronavirus or rhinovirus (Extended Data Fig. 9). In univariable analyses of factors associated with detection of respiratory viruses in various sample types, we did not identify significant association in viral shedding with days since symptom onset (Supplementary Table 3) for respiratory droplets or aerosols (Supplementary Tables 4–6).A subset of participants (72 of 246, 29%) did not cough at all during at least one exhaled breath collection, including 37 of 147 (25%) during the without-mask and 42 of 148 (28%) during the with-mask breath collection. In the subset for coronavirus (n = 4), we did not detect any virus in respiratory droplets or aerosols from any participants. In the subset for influenza virus (n = 9), we detected virus in aerosols but not respiratory droplets from one participant. In the subset for rhinovirus (n = 17), we detected virus in respiratory droplets from three participants, and we detected virus in aerosols in five participants.DiscussionOur results indicate that aerosol transmission is a potential mode of transmission for coronaviruses as well as influenza viruses and rhinoviruses. Published studies detected respiratory viruses13,14 such as influenza12,15 and rhinovirus16 from exhaled breath, and the detection of SARS-CoV17 and MERS-CoV18 from air samples (without size fractionation) collected from hospitals treating patients with severe acute respiratory syndrome and Middle East respiratory syndrome, but ours demonstrates detection of human seasonal coronaviruses in exhaled breath, including the detection of OC43 and HKU1 from respiratory droplets and NL63, OC43 and HKU1 from aerosols.Our findings indicate that surgical masks can efficaciously reduce the emission of influenza virus particles into the environment in respiratory droplets, but not in aerosols12. Both the previous and current study used a bioaerosol collecting device, the Gesundheit-II (G-II)12,15,19, to capture exhaled breath particles and differentiated them into two size fractions, where exhaled breath coarse particles >5 μm (respiratory droplets) were collected by impaction with a 5-μm slit inertial Teflon impactor and the remaining fine particles ≤5 μm (aerosols) were collected by condensation in buffer. We also demonstrated the efficacy of surgical masks to reduce coronavirus detection and viral copies in large respiratory droplets and in aerosols (Table 1b). This has important implications for control of COVID-19, suggesting that surgical face masks could be used by ill people to reduce onward transmission.Among the samples collected without a face mask, we found that the majority of participants with influenza virus and coronavirus infection did not shed detectable virus in respiratory droplets or aerosols, whereas for rhinovirus we detected virus in aerosols in 19 of 34 (56%) participants (compared to 4 of 10 (40%) for influenza and 8 of 23 (35%) for coronavirus). For those who did shed virus in respiratory droplets and aerosols, viral load in both tended to be low (Fig. 1). Given the high collection efficiency of the G-II (ref. 19) and given that each exhaled breath collection was conducted for 30 min, this might imply that prolonged close contact would be required for transmission to occur, even if transmission was primarily via aerosols, as has been described for rhinovirus colds20. Our results also indicate that there could be considerable heterogeneity in contagiousness of individuals with coronavirus and influenza virus infections.The major limitation of our study was the large proportion of participants with undetectable viral shedding in exhaled breath for each of the viruses studied. We could have increased the sampling duration beyond 30 min to increase the viral shedding being captured, at the cost of acceptability in some participants. An alternative approach would be to invite participants to perform forced coughs during exhaled breath collection12. However, it was the aim of our present study to focus on recovering respiratory virus in exhaled breath in a real-life situation and we expected that some individuals during an acute respiratory illness would not cough much or at all. Indeed, we identified virus RNA in a small number of participants who did not cough at all during the 30-min exhaled breath collection, which would suggest droplet and aerosol routes of transmission are possible from individuals with no obvious signs or symptoms. Another limitation is that we did not confirm the infectivity of coronavirus or rhinovirus detected in exhaled breath. While the G-II was designed to preserve viability of viruses in aerosols, and in the present study we were able to identify infectious influenza virus in aerosols, we did not attempt to culture coronavirus or rhinovirus from the corresponding aerosol samples.MethodsStudy designParticipants were recruited year-round from March 2013 through May 2016 in a general outpatient clinic of a private hospital in Hong Kong. As routine practice, clinic staff screened all individuals attending the clinics for respiratory and any other symptoms regardless of the purpose of the visit at triage. Study staff then approached immediately those who reported at least one of the following symptoms of ARI for further screening: fever ≥37.8 °C, cough, sore throat, runny nose, headache, myalgia and phlegm. Individuals who reported ≥2 ARI symptoms, within 3 d of illness onset and ≥11 years of age were eligible to participate. After explaining the study to and obtaining informed consent from the participants, a rapid influenza diagnostic test, the Sofia Influenza A + B Fluorescent Immunoassay Analyzer (cat. no. 20218, Quidel), was used to identify influenza A or B virus infection as an incentive to participate. All participants provided a nasal swab for the rapid test and an additional nasal swab and a separate throat swab for subsequent virologic confirmation at the laboratory. All participants also completed a questionnaire to record basic information including age, sex, symptom severity, medication, medical conditions and smoking history. In the first phase of the study from March 2013 to February 2014 (‘Influenza Study’), the result of the rapid test was used to determine eligibility for further participation in the study and exhaled breath collection, whereas in the second phase of the study from March 2014 to May 2016 (‘Respiratory Virus Study’), the rapid test did not affect eligibility. Eligible participants were then invited to provide an exhaled breath sample for 30 min in the same clinic visit.Before exhaled breath collection, each participant was randomly allocated in a 1:1 ratio to either wearing a surgical face mask (cat. no. 62356, Kimberly-Clark) or not during the collection. To mimic the real-life situation, under observation by the study staff, participants were asked to attach the surgical mask themselves, but instruction on how to wear the mask properly was given when the participant wore the mask incorrectly. Participants were instructed to breathe as normal during the collection, but (natural) coughing was allowed and the number of coughs was recorded by study staff. Participants were then invited to provide a second exhaled breath sample of the alternate type (for example if the participant was first assigned to wearing a mask they would then provide a second sample without a mask), but most participants did not agree to stay for a second measurement because of time constraints. Participants were compensated for each 30-min exhaled breath collection with a supermarket coupon worth approximately US$30 and all participants were gifted a tympanic thermometer worth approximately US$20.Ethical approvalWritten informed consent was obtained from all participants ≥18 years of age and written informed consent was obtained from parents or legal guardians of participants 11–17 years of age in addition to their own written informed consent. The study protocol was approved by the Institutional Review Board of The University of Hong Kong and the Clinical and Research Ethics Committee of Hong Kong Baptist Hospital.Collection of swabs and exhaled breath particlesNasal swabs and throat swabs were collected separately, placed in virus transport medium, stored and transported to the laboratory at 2–8 °C and the virus transport medium was aliquoted and stored at −70 °C until further analysis. Exhaled breath particles were captured and differentiated into two size fractions, the coarse fraction containing particles with aerodynamic diameter >5 μm (referred to here as ‘respiratory droplets’), which included droplets up to approximately 100 µm in diameter and the fine fraction with particles ≤5 μm (referred to here as ‘aerosols’) by the G-II bioaerosol collecting device12,15,19. In the G-II device, exhaled breath coarse particles >5 μm were collected by a 5-μm slit inertial Teflon impactor and the remaining fine particles ≤5 μm were condensed and collected into approximately 170 ml of 0.1% BSA/PBS. Both the impactor and the condensate were stored and transported to the laboratory at 2–8 °C. The virus on the impactor was recovered into 1 ml and the condensate was concentrated into 2 ml of 0.1% BSA/PBS, aliquoted and stored at −70 °C until further analysis. In a validation study, the G-II was able to recover over 85% of fine particles >0.05 µm in size and had comparable collection efficiency of influenza virus as the SKC BioSampler19.Laboratory testingSamples collected from the two studies were tested at the same time. Nasal swab samples were first tested by a diagnostic-use viral panel, xTAG Respiratory Viral Panel (Abbott Molecular) to qualitatively detect 12 common respiratory viruses and subtypes including coronaviruses (NL63, OC43, 229E and HKU1), influenza A (nonspecific, H1 and H3) and B viruses, respiratory syncytial virus, parainfluenza virus (types 1–4), adenovirus, human metapneumovirus and enterovirus/rhinovirus. After one or more of the candidate respiratory viruses was detected by the viral panel from the nasal swab, all the samples from the same participant (nasal swab, throat swab, respiratory droplets and aerosols) were then tested with RT–PCR specific for the candidate virus(es) for determination of virus concentration in the samples. Infectious influenza virus was identified by viral culture using MDCK cells as described previously21, whereas viral culture was not performed for coronavirus and rhinovirus.Statistical analysesThe primary outcome of the study was virus generation rate in tidal breathing of participants infected by different respiratory viruses and the efficacy of face masks in preventing virus dissemination in exhaled breath, separately considering the respiratory droplets and aerosols. The secondary outcomes were correlation between viral shedding in nose swabs, throat swabs, respiratory droplets and aerosols and factors affecting viral shedding in respiratory droplets and aerosols.We identified three groups of respiratory viruses with the highest frequency of infection as identified by RT–PCR, namely coronavirus (including NL63, OC43, HKU1 and 229E), influenza virus and rhinovirus, for further statistical analyses. We defined viral shedding as log10 virus copies per sample and plotted viral shedding in each sample (nasal swab, throat swab, respiratory droplets and aerosols); the latter two were stratified by mask intervention. As a proxy for the efficacy of face masks in preventing transmission of respiratory viruses via respiratory droplet and aerosol routes, we compared the respiratory virus viral shedding in respiratory droplet and aerosol samples between participants wearing face masks or not, by comparing the frequency of detection with a two-sided Fisher’s exact test and by comparing viral load (defined as log10 virus copies per sample) by an unadjusted univariate Tobit regression model, which allowed for censoring at the lower limit of detection of the RT–PCR assay. We also used the unadjusted univariate Tobit regression to investigate factors affecting viral shedding in respiratory droplets and aerosols without mask use, for example age, days since symptom onset, previous influenza vaccination, current medication and number of coughs during exhaled breath collection. We investigated correlations between viral shedding in nasal swab, throat swab, respiratory droplets and aerosols with scatter-plots and calculated the Spearman’s rank correlation coefficient between any two types of samples. We imputed 0.3 log10 virus copies ml−1 for undetectable values before transformation to log10 virus copies per sample. All analyses were conducted with R v.3.6.0 (ref. 22) and the VGAM package v.1.1.1 (ref. 23).Reporting SummaryFurther information on research design is available in the Nature Research Reporting Summary linked to this article.\n \n\n Data availabilityAnonymized raw data and R syntax to reproduce all the analyses, figures, tables and supplementary tables in the published article are available at: https://doi.org/10.5061/dryad.w9ghx3fkt.References1.Nichols, W. G., Peck Campbell, A. J. & Boeckh, M. Respiratory viruses other than influenza virus: impact and therapeutic advances. Clin. Microbiol. Rev. 21, 274–290 (2008).2.Shiu, E. Y. C., Leung, N. H. L. & Cowling, B. J. Controversy around airborne versus droplet transmission of respiratory viruses: implication for infection prevention. Curr. Opin. Infect. Dis. 32, 372–379 (2019).3.Tellier, R., Li, Y., Cowling, B. J. & Tang, J. W. Recognition of aerosol transmission of infectious agents: a commentary. BMC Infect. Dis. 19, 101 (2019).4.Xiao, J. et al. Nonpharmaceutical measures for pandemic influenza in nonhealthcare settings-personal protective and environmental measures. Emerg. Infect. Dis. https://doi.org/10.3201/eid2605.190994 (2020).5.Kutter, J. S., Spronken, M. I., Fraaij, P. L., Fouchier, R. A. M. & Herfst, S. Transmission routes of respiratory viruses among humans. Curr. Opin. Virol. 28, 142–151 (2018).6.Cowling, B. J. & Leung, G. M. Epidemiological research priorities for public health control of the ongoing global novel coronavirus (2019-nCoV) outbreak. Euro Surveill. https://doi.org/10.2807/1560-7917.ES.2020.25.6.2000110 (2020).7.Han, Q., Lin, Q., Ni, Z. & You, L. Uncertainties about the transmission routes of 2019 novel coronavirus. Influenza Other Respir. Viruses https://doi.org/10.1111/irv.12735 (2020).8.MacIntyre, C. R. & Chughtai, A. A. Facemasks for the prevention of infection in healthcare and community settings. BMJ 350, h694 (2015).9.Ha’eri, G. B. & Wiley, A. M. The efficacy of standard surgical face masks: an investigation using “tracer particles”. Clin. Orthop. Relat. Res. 148, 160–162 (1980).10.Patel, R. B., Skaria, S. D., Mansour, M. M. & Smaldone, G. C. Respiratory source control using a surgical mask: an in vitro study. J. Occup. Environ. Hyg. 13, 569–576 (2016).11.Johnson, D. F., Druce, J. D., Birch, C. & Grayson, M. L. A quantitative assessment of the efficacy of surgical and N95 masks to filter influenza virus in patients with acute influenza infection. Clin. Infect. Dis. 49, 275–277 (2009).12.Milton, D. K., Fabian, M. P., Cowling, B. J., Grantham, M. L. & McDevitt, J. J. Influenza virus aerosols in human exhaled breath: particle size, culturability, and effect of surgical masks. PLoS Pathog. 9, e1003205 (2013).13.Huynh, K. N., Oliver, B. G., Stelzer, S., Rawlinson, W. D. & Tovey, E. R. A new method for sampling and detection of exhaled respiratory virus aerosols. Clin. Infect. Dis. 46, 93–95 (2008).14.Stelzer-Braid, S. et al. Exhalation of respiratory viruses by breathing, coughing and talking. J. Med. Virol. 81, 1674–1679 (2009).15.Yan, J. et al. Infectious virus in exhaled breath of symptomatic seasonal influenza cases from a college community. Proc. Natl Acad. Sci. USA 115, 1081–1086 (2018).16.Tovey, E. R. et al. Rhinoviruses significantly affect day-to-day respiratory symptoms of children with asthma. J. Allergy Clin. Immunol. 135, 663–669 (2015).17.Booth, T. F. et al. Detection of airborne severe acute respiratory syndrome (SARS) coronavirus and environmental contamination in SARS outbreak units. J. Infect. Dis. 191, 1472–1477 (2005).18.Kim, S. H. et al. Extensive viable Middle East respiratory syndrome (MERS) coronavirus contamination in air and surrounding environment in MERS isolation wards. Clin. Infect. Dis. 63, 363–369 (2016).19.McDevitt, J. J. et al. Development and performance evaluation of an exhaled-breath bioaerosol collector for influenza virus. Aerosol Sci. Technol. 47, 444–451 (2013).20.Jennings, L. C. & Dick, E. C. Transmission and control of rhinovirus colds. Eur. J. Epidemiol. 3, 327–335 (1987).21.Chan, K. H., Peiris, J. S., Lim, W., Nicholls, J. M. & Chiu, S. S. Comparison of nasopharyngeal flocked swabs and aspirates for rapid diagnosis of respiratory viruses in children. J. Clin. Virol. 42, 65–69 (2008).22.R: a language and environment for statistical computing (R Foundation for Statistical Computing, Vienna, Austria, 2019).23.Yee, T. W. Vector Generalized Linear and Additive Models: with an Implementation in R (Springer, 2016).Download referencesAcknowledgementsThis work was supported by the General Research Fund of the University Grants Committee (grant no. 765811), the Health and Medical Research Fund (grant no. 13120592) and a commissioned grant of the Food and Health Bureau and the Theme-based Research Scheme (project no. T11-705/14-N) of the Research Grants Council of the Hong Kong SAR Government. We acknowledge colleagues including R. O. P. Fung, A. K. W. Li, T. W. Y. Ng, T. H. C. So, P. Wu and Y. Xie for technical support in preparing and conducting this study and enrolling participants; J. K. M. Chan, S. Y. Ho, Y. Z. Liu and A. Yu for laboratory support; S. Ferguson, W. K. Leung, J. Pantelic, J. Wei and M. Wolfson for technical support in constructing and maintaining the G-II device; V. J. Fang, L. M. Ho and T. T. K. Lui for setting up the database; and C. W. Y. Cheung, L. F. K. Cheung, P. T. Y. Ching, A. C. H. Lai, D. W. Y. Lam, S. S. Y. Lo, A. S. K. Luk and other colleagues at the Outpatient Center and Infection Control Team of Hong Kong Baptist Hospital for facilitating this study.Author informationAuthor notesThese authors jointly supervised this work: Donald K. Milton, Benjamin J. Cowling.AffiliationsWHO Collaborating Centre for Infectious Disease Epidemiology and Control, School of Public Health, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, ChinaNancy H. L. Leung, Daniel K. W. Chu, Eunice Y. C. Shiu, Benien J. P. Hau, Hui-Ling Yen, Dennis K. M. Ip, J. S. Malik Peiris, Wing-Hong Seto, Gabriel M. Leung & Benjamin J. CowlingDepartment of Microbiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, ChinaKwok-Hung ChanDepartment of Environmental Health, Harvard School of Public Health, Boston, MA, USAJames J. McDevittDepartment of Surgery, Queen Mary Hospital, Hong Kong, ChinaBenien J. P. HauDepartment of Mechanical Engineering, The University of Hong Kong, Hong Kong, ChinaYuguo LiDepartment of Pathology, Hong Kong Baptist Hospital, Hong Kong, ChinaWing-Hong SetoMaryland Institute for Applied Environmental Health, School of Public Health, University of Maryland, College Park, MD, USADonald K. MiltonAuthorsNancy H. L. LeungYou can also search for this author in Daniel K. W. ChuYou can also search for this author in Eunice Y. C. ShiuYou can also search for this author in Kwok-Hung ChanYou can also search for this author in James J. McDevittYou can also search for this author in Benien J. P. HauYou can also search for this author in Hui-Ling YenYou can also search for this author in Yuguo LiYou can also search for this author in Dennis K. M. IpYou can also search for this author in J. S. Malik PeirisYou can also search for this author in Wing-Hong SetoYou can also search for this author in Gabriel M. LeungYou can also search for this author in Donald K. MiltonYou can also search for this author in Benjamin J. CowlingYou can also search for this author in ContributionsAll authors meet the International Committee of Medical Journal Editors criteria for authorship. The study protocol was drafted by N.H.L.L. and B.J.C. Data were collected by N.H.L.L., E.Y.C.S. and B.J.P.H. Laboratory testing was performed by D.K.W.C. and K.-H.C. Statistical analyses were conducted by N.H.L.L. N.H.L.L. and B.J.C. wrote the first draft of the manuscript, and all authors provided critical review and revision of the text and approved the final version.Corresponding authorCorrespondence to\n Benjamin J. Cowling.Ethics declarations\n \n Competing interests\n B.J.C. consults for Roche and Sanofi Pasteur. The authors declare no other competing interests.\n \n Additional informationPeer review information Alison Farrell was the primary editor on this article and managed its editorial process and peer review in collaboration with the rest of the editorial team.Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.Extended dataExtended Data Fig. 1Participant enrolment, randomization of mask intervention and identification of respiratory virus infection.Extended Data Fig. 2 Weekly number of respiratory virus infections identified by RT-PCR in symptomatic individuals who had provided exhaled breath samples (respiratory droplets and aerosols) during the study period.Blue, coronavirus; red, influenza virus; yellow, rhinovirus; green, other respiratory viruses including human metapneumovirus, parainfluenza virus, respiratory syncytial virus and adenovirus; white, no respiratory virus infection identified.Extended Data Fig. 3 Respiratory virus shedding in (a) nasal swab, (b) throat swab, (c) respiratory droplets and (d) aerosols in symptomatic individuals with coronavirus NL63, coronavirus OC43, coronavirus HKU1, influenza A and influenza B virus infection.For nasal swabs and throat swabs, all infected individuals identified by RT-PCR in any collected samples were included: coronavirus NL63 (n = 8), coronavirus OC43 (n = 5), coronavirus HKU1 (n = 4), influenza A virus (n = 31) and influenza B virus (n = 14). For respiratory droplets and aerosols, only infected individuals who provided exhaled breath samples while not wearing a surgical face mask were included: coronavirus NL63 (n = 3), coronavirus OC43 (n = 3), coronavirus HKU1 (n = 4), influenza A virus (n = 19) and influenza B virus (n = 6). The box plots indicate the median with the interquartile range (lower and upper hinge) and ± 1.5 × interquartile range from the first and third quartile (lower and upper whisker). Dark blue, coronavirus NL63; light blue, coronavirus OC43; brown, coronavirus HKU1; red, influenza A virus; orange, influenza B virus.Extended Data Fig. 4 Efficacy of surgical face masks in reducing respiratory virus shedding in respiratory droplets and aerosols of symptomatic individuals with seasonal coronaviruses including (a) coronavirus NL63, (b) coronavirus OC43 and (c) coronavirus HKU1.The figure shows the virus copies per sample collected in nasal swab (red), throat swab (blue), respiratory droplets collected for 30 min while not wearing (dark green) or wearing (light green) a surgical face mask and aerosols collected for 30 min while not wearing (brown) or wearing (orange) a face mask, collected from individuals with acute respiratory symptoms who were positive for coronavirus NL63, coronavirus OC43 and coronavirus HKU1 as determined by RT-PCR in any samples. P values for mask intervention as predictor of log10 virus copies per sample in an unadjusted univariate Tobit regression model which allowed for censoring at the lower limit of detection of the RT-PCR assay are shown, with significant differences in bold. For nasal swabs and throat swabs, all infected individuals were included (coronavirus NL63, n = 8; coronavirus OC43, n = 5; coronavirus HKU1, n = 4). For respiratory droplets and aerosols, numbers of infected individuals who provided exhaled breath samples while not wearing or wearing a surgical face mask, respectively were: coronavirus NL63 (n = 3 and 5), coronavirus OC43 (n = 3 and 4), coronavirus HKU1 (n = 4 and 2). A subset of participants provided exhaled breath samples for both mask interventions (coronavirus NL63, n = 0; coronavirus OC43, n = 2; coronavirus HKU1, n = 2).Extended Data Fig. 5 Efficacy of surgical face masks in reducing respiratory virus shedding in respiratory droplets and aerosols of symptomatic individuals with seasonal influenza viruses including (a) influenza A and (b) influenza B virus.The figure shows the virus copies per sample collected in nasal swab (red), throat swab (blue), respiratory droplets collected for 30 min while not wearing (dark green) or wearing (light green) a surgical face mask and aerosols collected for 30 min while not wearing (brown) or wearing (orange) a face mask, collected from individuals with acute respiratory symptoms who were positive for influenza A and influenza B virus as determined by RT-PCR in any samples. P values for mask intervention as predictor of log10 virus copies per sample in an unadjusted univariate Tobit regression model which allowed for censoring at the lower limit of detection of the RT-PCR assay are shown, with significant differences in bold. For nasal swabs and throat swabs, all infected individuals were included (influenza A virus, n = 31; influenza B virus, n = 14). For respiratory droplets and aerosols, numbers of infected individuals who provided exhaled breath samples while not wearing or wearing a surgical face mask, respectively were: influenza A virus (n = 19 and 19), influenza B virus (n = 6 and 10). A subset of participants provided exhaled breath samples for both mask interventions (influenza A virus, n = 7; influenza B virus, n = 2).Extended Data Fig. 6 Correlation of coronavirus viral shedding between different samples (nasal swab, throat swab, respiratory droplets and aerosols) in symptomatic individuals with seasonal coronavirus infection.For nasal swabs and throat swabs, all infected individuals were included (n = 17). For respiratory droplets and aerosols, only infected individuals who provided exhaled breath samples while not wearing a surgical face mask were included (n = 10). r, the Spearman’s rank correlation coefficient.Extended Data Fig. 7 Correlation of influenza viral shedding between different samples (nasal swab, throat swab, respiratory droplets and aerosols) in symptomatic individuals with seasonal influenza infection.For nasal swabs and throat swabs, all infected individuals were included (n = 43). For respiratory droplets and aerosols, only infected individuals who provided exhaled breath samples while not wearing a surgical face mask were included (n = 23). r, the Spearman’s rank correlation coefficient.Extended Data Fig. 8 Correlation of rhinovirus viral shedding between different samples (nasal swab, throat swab, respiratory droplets and aerosols) in symptomatic individuals with rhinovirus infection.For nasal swabs and throat swabs, all infected individuals were included (n = 54). For respiratory droplets and aerosols, only infected individuals who provided exhaled breath samples while not wearing a surgical face mask were included (n = 36). r, the Spearman’s rank correlation coefficient.Extended Data Fig. 9 Respiratory virus shedding in respiratory droplets and aerosols stratified by days from symptom onset for (a) coronavirus, (b) influenza virus or (c) rhinovirus.The figures shows the virus copies per sample collected in nasal swab (red), throat swab (blue), respiratory droplets (dark green) and aerosols (brown) collected for 30 min while not wearing a surgical face mask, stratified by the number of days from symptom onset on which the respiratory droplets and aerosols were collected. For nasal swabs and throat swabs, all infected individuals were included (coronavirus, n = 17; influenza virus, n = 43; rhinovirus, n = 54). For respiratory droplets and aerosols, numbers of infected individuals who provided exhaled breath samples while not wearing or wearing a surgical face mask, respectively were: coronavirus (n = 10 and 11), influenza virus (n = 23 and 28), rhinovirus (n = 36 and 32). A subset of participants provided exhaled breath samples for both mask interventions (coronavirus, n = 4; influenza virus, n = 8; rhinovirus, n = 14). The box plots indicate the median with the interquartile range (lower and upper hinge) and ± 1.5 × interquartile range from the first and third quartile (lower and upper whisker).Supplementary informationAbout this articleCite this articleLeung, N.H.L., Chu, D.K.W., Shiu, E.Y.C. et al. Respiratory virus shedding in exhaled breath and efficacy of face masks.\n Nat Med (2020). https://doi.org/10.1038/s41591-020-0843-2Download citationReceived: 02 March 2020Accepted: 20 March 2020Published: 03 April 2020DOI: https://doi.org/10.1038/s41591-020-0843-2"],["topImageUrl",null],["error",null]]},{"__iterable":"Map","data":[["title","職安系副教授實測「乾蒸口罩」5次內過濾效力仍合格，但強調僅應急不建議長期使用 - The News Lens 關鍵評論網"],["url","https://www.thenewslens.com/article/131064"],["summary","武漢肺炎疫情導致國內口罩需求量增大，網路上也出現各種可以讓口罩重複使用的「復活偏方」，包括用電鍋乾蒸、用紫外線照、用酒精消毒等。如今，中山醫學大學職安系副教授賴全裕表示，經過檢測證實，「電鍋乾蒸」口罩，5次以內，過濾效果都還在規定合格的80%以內。但中央流行疫情指揮中心強調，以台灣疫情狀況，非慢性病患，在開放空間不用戴口罩。《中央社》報導，賴全裕表示，「電鍋乾蒸法」可使口罩復活。電鍋乾蒸法可以利用一般家用電鍋、電鍋架及內鍋，在電鍋內依序置入內鍋架、內鍋，並放入沒沾上體液、血液或鼻涕的口罩，不用加水，直接按下煮飯按鍵，乾蒸約2到3分鐘，等到煮飯鍵跳起後，再繼續保溫約5分鐘，即可拿出來繼續使用。賴全裕指出，一般經認證合格的外科口罩，過濾效力約為80%，乾蒸5次後，過濾效力減損到70%以下，這時就必須丟棄，不能再使用了。但賴全裕也提醒，這是口罩短缺時的應急措施，不建議長期使用。根據中山醫學大學特色領域研究中心介紹，賴全裕研究專長為個人防護具（包含呼吸防護具、防護衣等）進行生物防護性能及消毒、再利用。此外，《自由時報》訪問亞東紀念醫院感染管制中心主任廖俊星，廖俊星表示，紫外線消毒、水蒸口罩、酒精消毒，都沒有研究證實能有效消毒口罩，其中，水蒸、酒精消毒還可能破壞口罩結構。但曾有人拿過期口罩測驗，如果口罩沒有被破壞或使用，證實效果還在。此外，昨日，中央流行疫情指揮中心的記者會上，也有記者提問表示，有台大醫師認為口罩用越久，效果越好。指揮中心醫療整備官羅一鈞澄清，這有違一般對於口罩的學理跟常識，有待證實。指揮中心昨日也再次呼籲，開放空間不用戴口罩，但去醫院看病、出入人多擁擠地方、慢性病患要戴口罩。此外，也要以肥皂肥皂勤洗手、避免觸摸眼鼻口。 \t \t\t \t\t\t消毒方法 \t\t\t是否證實有效 \t\t\t資訊來源 \t\t \t\t \t\t\t電鍋乾蒸 \t\t\t是 \t\t\t（乾蒸口罩超過5次即需丟棄） \t\t\t中山醫學大學職安系副教授賴全裕 \t\t \t\t \t\t\t紫外線消毒 \t\t\t未證實 \t\t\t亞東紀念醫院感染管制中心主任廖俊星 \t\t \t\t \t\t\t水蒸 \t\t\t未證實 \t\t\t亞東紀念醫院感染管制中心主任廖俊星 \t\t \t\t \t\t\t酒精消毒 \t\t\t未證實 \t\t\t亞東紀念醫院感染管制中心主任廖俊星 \t\t \t\t \t\t\t過期口罩 \t\t\t是 \t\t\t亞東紀念醫院感染管制中心主任廖俊星 \t\t \t\t \t\t\t口罩用越久效果越好 \t\t\t未證實 \t\t\t中央流行疫情指揮中心醫療整備官羅一鈞 \t\t \t 延伸閱讀： \t【插畫】「武漢肺炎」來勢洶洶，口罩該買哪種？ \t口罩實名制上路：如何買？怎麼查哪還有庫存？8個QA一次看懂 \t其實你不一定要戴口罩：武漢肺炎讓全台陷「口罩之亂」，6個QA看怎麼選、如何用新聞來源： \t口罩復活術真有效？醫師破解防疫網路謠言（自由時報） \t口罩電鍋乾蒸3分鐘殺菌學者實測最多重複用5次（中央社）責任編輯：李修慧"],["topImageUrl",null],["error",null]]},{"__iterable":"Map","data":[["title","認證電鍋可以乾蒸口罩！指揮中心︰殺菌實驗達標 - 生活 - 自由時報電子報"],["url","https://news.ltn.com.tw/news/life/breakingnews/3123669"],["summary","針對口罩使用電鍋乾蒸延長使用壽命進行實驗，結果發現無論細菌過濾效率或微粒過濾效率都跟原本口罩效果相近。（指揮中心提供）\n \n\n\n 2020-04-05 16:45:51〔記者簡惠茹／台北報導〕衛福部食藥署和中山醫學大學職業安全衛生系副教授賴全裕都針對口罩使用電鍋乾蒸延長使用壽命進行實驗，實驗結果發現無論細菌過濾效率或微粒過濾效率都跟原本口罩效果相近。指揮官陳時中指出，從這禮拜四4月9日開始，口罩就可以14天買9片，這個量應該是足夠，還是非常期待大家撙節使用，食藥署也有提供增加口罩使用次數或延長使用時間的方法。 \n \n食藥署說明電鍋乾蒸方法，一，先將內鍋架及內鍋依序放入乾燥電鍋；二、將醫用口罩平放於內鍋中，蓋上外鍋蓋；三、按下電鍋加熱開關，等約3分鐘待開關自動跳起後，再靜置5分鐘，使鍋內熱空氣充分殺菌；四、打開外鍋蓋，完成殺菌程序，待電鍋冷卻後再進行下一次口罩殺菌。指揮中心研究組副組長吳秀梅說明，自己使用過的醫用外科口罩如果想要延長壽命，可以放入電鍋內鍋架乾蒸，提醒不要放水，放水會影響口罩靜電層，有人問是否可以使用酒精消毒，其實也不好，因為靜電層會受到影響。吳秀梅也提醒，如果口罩有髒汙破損不建議再度消毒使用，有呼吸道症狀、慢性疾病、出入醫院以一次使用為原則，低風險場所就會建議可以使用布口罩，有不織布層的布口罩，可以重複清洗效果也好，也可以使用電鍋乾蒸重複消毒4到5次。吳秀梅說明外科口罩測試部分，除了有參考學研單位建議，食藥署也做過研究，她表示，經過3到5次消毒後，對於細菌過濾效率（BFE）可以維持99%以上。中山醫學大學副教授賴全裕實驗室做過電鍋乾蒸口罩實驗，他指出，粒子過濾效果（PFE）檢測中，儀器會產生2奈米到2微米的微粒，其中99%是10奈米到770奈米的微粒，然後來看口罩可以過濾多少，總過濾效果如果超過95%，就是N95口罩，80%以上是外科手術口罩，電鍋乾蒸實驗結果發現，第一次乾蒸跟原本外科口罩過濾效果相比還能維持99%，只是纖維帶電還是有些微影響，但是消毒越多次，就會越下降，建議不要超過5次。至於病毒過濾效果，賴全裕說，雖然外科手術口罩沒有檢測病毒過濾效果，主要是防護衣才會測試，但是武漢肺炎病毒的大小大概是100奈米，所以檢測PFE就有將病毒尺寸涵蓋在內。賴全裕也解答民眾各種電鍋蒸口罩的疑慮。許多民眾詢問可以使用家裡的電鍋就好嗎？還是要買新的電鍋？賴全裕指出，由於內鍋壁165度左右，該殺死的細菌應該都殺死了，乾蒸氣溫大概110度左右，如果有疑慮可以洗鍋子。口罩氣味太重怎麼辦？可以加精油或酒精嗎？賴全裕表示，有化學物質的都會破壞口罩不織布纖維帶電，不建議這樣做，因為味道來源是口水和細菌繁殖的關係，殺死後理論上味道就會不見。用烤箱、氣炸鍋或微波爐可以嗎？賴全裕指出，這些家電如果可以控制溫度達110度，可以模擬電鍋溫度的話就可以嘗試，但是還是要實測才知道，微波爐可能會出現口罩燃燒要小心。使用負離子吹風機可以嗎？賴全裕表示，目前還沒有實驗檢測，暫時不建議。殺菌燈小型烘碗機可以嗎？賴全裕指出，如果紫外燈波長有220奈米到280奈米，可是口罩裡面有3層，紫外燈只能對上下兩面照射，不確定中間層的殺菌效果，使用的時候還是建議要注意，未來有機會會再做實驗。用煮的可以嗎？賴全裕說，還沒有實驗可以確認。暫時拿下來的口罩如何收納？賴全裕指出，建議把口罩放在通風乾燥處，拿下來的時候，不要使用完全密封的塑膠袋，會造成細菌容易繁殖，可以用塑膠夾或紙袋，避免手觸摸到口罩。相關新聞請見︰武漢肺炎懶人包》傳染途徑、預防措施看這裡「武漢肺炎專區」請點此，更多相關訊息，帶您第一手掌握。\n\n \n\n\n\n \n\n \n 不用抽不用搶現在用APP看新聞保證天天中獎　\n 點我下載APP　\n 按我看活動辦法\n\n \n \n\n\n\n 相關新聞\n \n \n\n\n\n\n\n\n\n\n \n \n\n\n\n\n\n\n\n\n\n\n LTN集點大放送\n \n \n \n \n \n \n\n\n\n\n\n網友回應"],["topImageUrl",null],["error",null]]},{"__iterable":"Map","data":[["title","Virological assessment of hospitalized patients with COVID-2019"],["url","https://www.nature.com/articles/s41586-020-2196-x"],["summary","AbstractCoronavirus disease 2019 (COVID-19) is an acute respiratory tract infection that emerged in late 20191,2. Initial outbreaks in China involved 13.8% cases with severe, and 6.1% with critical courses3. This severe presentation corresponds to the usage of a virus receptor that is expressed predominantly in the lung2,4. By causing an early onset of severe symptoms, this same receptor tropism is thought to have determined pathogenicity, but also aided the control, of severe acute respiratory syndrome (SARS) in 20035. However, there are reports of COVID-19 cases with mild upper respiratory tract symptoms, suggesting the potential for pre- or oligosymptomatic transmission6–8. There is an urgent need for information on body site-specific virus replication, immunity, and infectivity. Here we provide a detailed virological analysis of nine cases, providing proof of active virus replication in upper respiratory tract tissues. Pharyngeal virus shedding was very high during the first week of symptoms (peak at 7.11 × 108 RNA copies per throat swab, day 4). Infectious virus was readily isolated from throat- and lung-derived samples, but not from stool samples, in spite of high virus RNA concentration. Blood and urine never yielded virus. Active replication in the throat was confirmed by viral replicative RNA intermediates in throat samples. Sequence-distinct virus populations were consistently detected in throat and lung samples from the same patient, proving independent replication. Shedding of viral RNA from sputum outlasted the end of symptoms. Seroconversion occurred after 7 days in 50% of patients (14 days in all), but was not followed by a rapid decline in viral load. COVID-19 can present as a mild upper respiratory tract illness. Active virus replication in the upper respiratory tract puts the prospects of COVID-19 containment in perspective.\n \n \n \n\n \n \n \n \n\n Author informationAuthor notesThese authors contributed equally: Roman Wölfel, Victor M. Corman, Wolfgang GuggemosThese authors jointly supervised this work: Christian Drosten, Clemens WendtnerAffiliationsBundeswehr Institute of Microbiology, Munich, GermanyRoman Wölfel, Sabine Zange, Patrick Vollmar, Rosina Ehmann & Katrin ZwirglmaierCharité Universitätsmedizin Berlin, Berlin, GermanyVictor M. Corman, Marcel A. Müller, Daniela Niemeyer, Terry C. Jones, Tobias Bleicker, Sebastian Brünink, Julia Schneider & Christian DrostenKlinikum München-Schwabing, Munich, GermanyWolfgang Guggemos, Michael Seilmaier & Clemens WendtnerCenter for Pathogen Evolution, Department of Zoology, University of Cambridge, Cambridge, UKTerry C. JonesUniversity Hospital LMU Munich, Munich, GermanyCamilla Rothe & Michael HoelscherAuthorsRoman WölfelYou can also search for this author in Victor M. CormanYou can also search for this author in Wolfgang GuggemosYou can also search for this author in Michael SeilmaierYou can also search for this author in Sabine ZangeYou can also search for this author in Marcel A. MüllerYou can also search for this author in Daniela NiemeyerYou can also search for this author in Terry C. JonesYou can also search for this author in Patrick VollmarYou can also search for this author in Camilla RotheYou can also search for this author in Michael HoelscherYou can also search for this author in Tobias BleickerYou can also search for this author in Sebastian BrüninkYou can also search for this author in Julia SchneiderYou can also search for this author in Rosina EhmannYou can also search for this author in Katrin ZwirglmaierYou can also search for this author in Christian DrostenYou can also search for this author in Clemens WendtnerYou can also search for this author in Corresponding authorsCorrespondence to\n Christian Drosten or Clemens Wendtner.Supplementary information\n \n \n Supplementary InformationThis file contains Supplementary Methods, which include a description of RNA extraction and RT-PCR methods, a description of cell culture and antibody detection methods and Supplementary References.About this articleCite this articleWölfel, R., Corman, V.M., Guggemos, W. et al. Virological assessment of hospitalized patients with COVID-2019.\n Nature (2020). https://doi.org/10.1038/s41586-020-2196-xDownload citationReceived: 01 March 2020Accepted: 24 March 2020Published: 01 April 2020DOI: https://doi.org/10.1038/s41586-020-2196-x\n\n \n\n \n CommentsBy submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate."],["topImageUrl",null],["error",null]]}]}],["text","#口罩可以乾蒸嗎\n#為何2月不行現在可以\n#水鏡來告訴你\n\n今天疾管中心宣傳口罩可以乾蒸，再重複使用\n馬上有人找出2月衛福部食藥署建議不要乾蒸的資料\n還有人把這照片放在爆料公社，很明顯要帶風向\n\n有人酸因為唐鳳說可以乾蒸，所以政策就轉彎\n\n不管別人怎麼說，包括這篇都是\n請做個獨立思考的人\n\n#防疫本來就是與時俱進\n大家應該還記得，2月的時候疾管中心跟大家說不用戴口罩\n總統、行政院長、陳時中部長，也帶頭不戴口罩\n當時還有「我OK，你先戴」的風氣\n\n當時背景是沒有社區感染，戴不戴口罩風險都低\n當時到現在沒戴口罩的人，都活得好好的也沒被感染。\n\n這兩天陳時中部長已經戴起口罩，為什麼呢？\n簡單說，我們有零星的社區感染，加上這幾天連假的出遊\n我們要在未來的14天防範疫情社區化\n\nNature Medicine刊出香港學者研究，外科口罩可以顯著防止新冠病毒從感染者傳播出去。https://www.nature.com/articles/s41591-020-0843-2?fbclid=IwAR3EzP6Xm2bXH4bDHx9i9oOYxqHIraWbuRbZcGzM6fmgQz7iIl7ufHrkN-A\n\n這是因為疫情的不同，資訊的改變\n政府反應以及需要民間配合的方式不同\n\n#那到底口罩可不可以乾蒸\n\n你可能會問，當時食藥署為何覺得不行？\n當時食藥署的說法是\"可能\"會破壞結構，所以不建議。\n\n但後續的研究出來，真的可以透過乾蒸讓口罩重複使用\n\n根據中山醫學大學職安系副教授賴全裕表示，經過檢測證實，「電鍋乾蒸」口罩，5次以內，過濾效果都還在規定合格的80%以內。\nhttps://www.thenewslens.com/article/131064\n\n實驗結果發現無論細菌過濾效率或微粒過濾效率都跟原本口罩效果相近。\nhttps://news.ltn.com.tw/news/life/breakingnews/3123669\n\n再說一次，這是人類第一次遇到這個病毒，很多事情本來就不了解。\n\n比如Nature這篇研究，就刷新大家原本的認知\n\n原來剛被感染的前5天，傳染力最大，到住院第9天傳染力就低了，不一定要在負壓房。\n\n這對本來以為症狀越嚴重傳播力越大、負壓房要住好住滿的觀念是很大的釐清\nhttps://www.nature.com/articles/s41586-020-2196-x\n\n好，回到乾蒸口罩這件事\n原先政府有疑惑，後來研究證實，口罩乾蒸可以重複使用\n政府提供資訊，不OK嗎？\n\n#未來14天大家照顧好自己\n\n現在已經有人在傳\n「為了要向國外捐口罩，所以要求民眾乾蒸」\n\n請問我們有為了捐口罩，一周從買三個變買兩個嗎？\n\n提供乾蒸，只是資訊更新後提供方法\n可能有些人不覺得一個口罩五元貴\n\n但對節省、重視環保的或有經濟壓力的人，這不失為權宜之計\n至於要酸的、要戰的、要攻擊的，\n就別了，台灣需要大家的團結\n\n就像這幾天的連假，\n大家玩都玩了，該得的也得了，不該得的也不會得\n\n本院只有一個請求\n\n#連假有出遊的朋友接下來14天外出請戴口罩\n\n接下來14天，大家把自己照顧好\n勤洗手、出門戴口罩、保持人際距離、不摸臉、少聚集\n\n#祝大家平安健康\n\n#我們二月也不知道肥胖容易得重症啊\n\n#水鏡共筆"],["id","1kyvy3pelrgow"],["createdAt","2020-04-06T08:20:58.169Z"]]},{"__iterable":"Map","data":[["replyCount",1],["references",{"__iterable":"List","data":[{"__iterable":"Map","data":[["type","LINE"]]}]}],["replyRequestCount",1],["user",{"__iterable":"Map","data":[["id","j4S8C_AO7U9UgO3jfO7w-_TCIO4Xtij_63hrFiTOAIF-wiO0o"],["name","忠誠的✖長濱✖法蘭克林"],["avatarUrl",null],["level",0]]}],["hyperlinks",{"__iterable":"List","data":[{"__iterable":"Map","data":[["title","Universal and reusable virus deactivation system for respiratory protection"],["url","https://www.nature.com/articles/srep39956#Sec8"],["summary","AbstractAerosolized pathogens are a leading cause of respiratory infection and transmission. Currently used protective measures pose potential risk of primary/secondary infection and transmission. Here, we report the development of a universal, reusable virus deactivation system by functionalization of the main fibrous filtration unit of surgical mask with sodium chloride salt. The salt coating on the fiber surface dissolves upon exposure to virus aerosols and recrystallizes during drying, destroying the pathogens. When tested with tightly sealed sides, salt-coated filters showed remarkably higher filtration efficiency than conventional mask filtration layer, and 100% survival rate was observed in mice infected with virus penetrated through salt-coated filters. Viruses captured on salt-coated filters exhibited rapid infectivity loss compared to gradual decrease on bare filters. Salt-coated filters proved highly effective in deactivating influenza viruses regardless of subtypes and following storage in harsh environmental conditions. Our results can be applied in obtaining a broad-spectrum, airborne pathogen prevention device in preparation for epidemic and pandemic of respiratory diseases.\n \n \n \n \n \n \n\n \n IntroductionAerosols take a prominent role in airborne transmission of respiratory diseases. Droplets with aerodynamic size (da) < 10 μm and 10 < da < 100 μm are known to infect the alveolar regions and upper respiratory tract, respectively1,2. Notably, aerosols can also be a route of infection in diseases that, contrary to for instance influenza, do not specifically target the respiratory tract, as it could be the case of Ebola virus3. While vaccination can greatly reduce morbidity and mortality, during a pandemic or epidemic new vaccines matching the specific strain would be available, at the earliest, six months after the initial outbreak. Additionally, following development of an effective viral vaccine, several potential problems would remain, such as limited supply due to insufficient production capacity and time-consuming manufacturing processes. As a result, individuals close to the point of an outbreak would be in imminent danger of exposure to infectious diseases during the non-vaccine period. In the absence of vaccination, respirators and masks can be worn to prevent transmission of airborne pathogenic aerosols and control diseases, such as influenza4.The main alternative, the N95 respirator, requires training prior to use, must be expertly fitted to address the risk of faceseal leakage at the face-mask interface, and must be disposed of as biohazard5. Due to these factors, the use of N95 respirators on a large scale is impractical and expensive during an epidemic or pandemic. Past experiences of severe acute respiratory syndrome (SARS), H1N1 swine flu in 2009, and Middle East respiratory syndrome (MERS) indicate that surgical masks have been most widely adopted by the public as personal protective measure, despite controversy on their effectiveness6,7,8,9. Currently, among other factors, filtration in respirators and masks depends on filter characteristics, including fiber diameter, packing density, charge of fibers and filter thickness, as well as particle properties, such as diameter, density and velocity10,11,12,13,14. However, in the lack of a system to deactivate the collected pathogens, safety concerns naturally arise about secondary infection and contamination from virus-laden filter media during utilization and disposal. Furthermore, since re-sterilization is not possible without causing damage, respirators and masks are recommended for single use only9,15,16. Scientific efforts have been focused on treatment of filters with materials possessing well-known antimicrobial properties, such as iodine, chlorine and metals17,18,19,20,21,22,23,24,25, although with limited effectiveness against virus aerosols26,27,28. Therefore, a key challenge is the development of an easy-to-use, universal virus negation system, which is reusable without reprocessing and capable of deactivating pathogens, thereby reducing potential risk of secondary infection and transmission.Here, we report a simple but efficient virus inactivation system exploiting the naturally occurring salt recrystallization. Our strategy is to modify the surface of the fibrous filtration layer within masks with a continuous salt film for virus deactivation via two successive processes: i) salt is locally dissolved by the viral aerosols and ii) supersaturation is followed by evaporation-induced salt recrystallization. Consequently, viruses are exposed to increasingly higher concentrations of saline solution during drying and physically damaged by recrystallization.ResultsPreparation and characterization of salt-functionalized filtersTo demonstrate the concept of virus deactivation system based on salt recrystallization, the middle layer of three-ply surgical mask, polypropylene (PP) microfiber filter, was coated with NaCl salt as an active virus negation unit (see Supplementary Fig. S1 for bare PP filter). The coating formulations contained surfactant to enhance wetting of saline solution on the surface of hydrophobic PP fibers. Bare PP filters (abbreviated as Filterbare) were pre-wet to contain about 600 μL of coating solution (abbreviated as Filterwet). The amount of NaCl salt (Wsalt in mg/cm2) coated on the filter per unit area, considering that the filters thickness is constant, was easily controlled by changing the coating solution volume (Vsalt in μL) during drying of pre-wet filter (radius: 3 cm, Wsalt = 3.011 + 0.013 × Vsat, n = 7) (Fig. S2). Scanning electron microscopy (SEM) and energy dispersive X-ray (EDX) mapping analysis showed the formation of homogeneous NaCl coating during drying, as also confirmed by X-ray diffraction (XRD) (Fig. 1a,b and Supplementary Fig. S3). Both the formation of NaCl coating on PP fibers and presence of surfactant in the coating formulation appeared to alter the filter surface properties from hydrophobic (bare filter; contact angle, θc = 133.0 ± 4.7°) to completely hydrophilic (salt-coated filter; θc ~ 0°, n = 10) (Fig. 1c and Supplementary Fig. S4). Hydrophilic nature of salt coating can greatly improve adhesion of viral aerosols to PP fibers compared to Filterbare, as seen in Raman microscope images (Fig. 1d and Supplementary Fig. S5).Figure 1: Mask with salt-coated filter for prevention and deactivation of airborne pathogens.(a) SEM image of Filterwet+600μL (top left) and EDX mapping images of Na (red), Cl (green), and NaCl (combination of Na and Cl mapping images), showing the formation of NaCl coating, as also confirmed by XRD spectra (b) of Filterbare, Filterwet, Filterwet+100μL, Filterwet+300μL, Filterwet+600μL, Filterwet+900μL and Filterwet+1200μL (labelled as Bare, wet, wet+100 μL, wet+300 μL, wet+600 μL, wet+900 μL and wet+1200 μL, respectively; miller indices corresponding to NaCl crystal are shown at the top of XRD spectra for each position). (c) Optical microscope images for contact angle measurements using 3 μL DI water droplets on (i) Filterbare and (ii) Filterwet+600μL (n = 10). (d) Microscope images of aerosol on (i) Filterbare and (ii) Filterwet+600μL (n = 10).Filtration efficiency against viral aerosols and protective efficacy in vivoFiltration efficiency of salt-coated filters was tested against aerosols with volumetric mean diameter (VMD) of 2.5–4 μm containing H1N1 pandemic influenza virus (A/California/04/2009, abbreviated as CA/09) at different pressure conditions (see Fig. 2a for transmission electron microscope (TEM) image of H1N1 virus). Interestingly, as shown in Fig. 2b, Filterbare did not exhibit any significant level of resistance against penetration of virus under our experimental conditions (i.e., 0% filtration efficiency). Conversely, salt-coated filters showed substantially increasing filtration efficiency with pressure and amount of coated salt. In particular, in the case of Filterwet+600μL, filtration efficiency varied from 43 to 70%, with increasing pressure from 3 to 17 kPa, and Filterwet+1200μL exhibited persistent, high-level efficiency (~85%) (one-way ANOVA, P = 0.85).Figure 2: Filtration efficiency of salt-coated filters.(a) TEM image of CA/09 H1N1 influenza virus. (b) Pressure-dependent filtration efficiency (n = 8–10, mean ± standard deviation (SD)). (c–f) Effects of filtration efficiency on protective efficacy in vivo. Body weight change of mice after infection with the dosages of penetrated virus (n = 12, mean ± SD) (c), survival rates (mean; 100% means that all mice in the group survived as penetrated dosages were lower than lethal dose) (d), lung virus titers (n = 4, mean ± SD) (e), and lung inflammatory cytokine (interferon-γ (IFN-γ)) assay (n = 11, mean ± SD) (f). Legends: filters are labelled as in Fig. 1b.To probe the effects of filtration efficiency on protective efficacy, in vivo experiments were performed using mice intranasally (IN) infected with penetrated dosages of H1N1 virus under breathing pressure (~10 kPa)29. As shown in Fig. 2c, similarly to negative control groups (mice infected with lethal dose of virus stock and aerosolized virus), mice exposed to a dose penetrated through the bare filter showed rapid body weight loss, followed by death within 10 days after infection, in good agreement with the observed 0% filtration efficiency (Fig. 2b). In contrast, mice groups exposed to virus derived from salt-coated filters resulted in 100% survival rate (Fig. 2d). Furthermore, lungs of mice from negative control groups exhibited severe lung infection 4 days after challenge (Fig. 2e). Conversely, mice groups exposed to virus derived from salt-coated filters showed significantly lower levels of lung viral titers (t-test, P < 0.005). This is consistent with lower levels of inflammatory cytokines, interferon-γ (IFN-γ), from salt-coated filter groups compared to negative control and bare filter groups (t-test, P < 0.001) (Fig. 2f).Deactivation of virus on salt-functionalized filtersInfluenza virus stability tests were performed to investigate the effects of salt coating. The same amount of recovered viruses from the PP fibers was used, and, in the case of bare filters, viral aerosols exposure was conducted in the absence of pressure due to 100% penetration of viral aerosols. Unlike bare filters (Fig. S6a(i)), formation of micron-sized NaCl phase represents a typical feature of salt-coated filters due to recrystallization of NaCl salt, following local dissolution upon aerosols exposure (SEM images in Fig. S6a, ii to iv, and EDX mapping in Fig. S6b). In contrast to 8% HA activity loss of virus adsorbed onto Filterbare, salt-coated filters exhibited almost complete HA activity loss within 5 min of incubation (Fig. 3a). Such dramatic virus destabilization on salt-coated filters is further supported by negligible levels of viral titers compared to Filterbare with incubation time (t-test, P < 0.001) (Fig. 3b). It is also noted that virus titers exhibited significant decrease with increase of incubation time and amount of coated salt (ANOVA general linear model, P < 0.001). TEM analysis showed that influenza virus on Filterbare experiences morphological change into non-spherical shape during aerosol drying (Fig. 3c(i)). Notably, influenza virus was severely damaged on salt-coated filters even at 5 min of incubation (Fig. 3c(ii)). From microscopic analysis, aerosol drying time was about 3 min, indicating that destruction of virus observed at 5 min is associated with drying-induced salt crystallization. Physical damage of virus due to crystallization was similarly reported as a major destabilizing factor of inactivated influenza virus30,31. Lower levels of native fluorescence and nile red fluorescence from virus recovered from salt-coated filters accounted for more severe conformational change of antigenic proteins and destabilization of viral envelope, respectively, consistent with TEM analysis (t-test, P < 0.001) (Fig. 3d). In parallel, we investigated the separate effect of salt concentration increase on virus stability during the aerosol drying process, irrespective of crystal growth. As displayed in Supplementary Fig. S7, the materials collected in suspension from Filterwet+600μL induced visible morphological transformation of the virus (Supplementary Fig. S7b) compared to suspension of Filterbare (Supplementary Fig. S7a). This can be attributed to the high salt/surfactant concentration and osmotic pressure, which have been well-known to destabilize proteins and viruses31,32,33. Therefore, the marked virus destabilization on salt-coated PP fibers can be explained by the combined effects of salt concentration increase during drying and evaporation-induced salt crystallization.Figure 3: Inactivation of virus adsorbed on salt-coated filters.(a,b) HA activity (a) and virus titer (b) displaying the effects of incubation time on the remaining activity of virus (n = 4–8, mean ± SD). (c) TEM images of viruses reconstituted, after incubation for 5 and 60 min, from (i) Filterbare and (ii) Filterwet+600μL. (d) Native fluorescence/nile red fluorescence of viruses incubated for 60 min (n = 12, mean ± SD). (e,f) Body weight change of mice after infection with virus recovered from filters after incubation for 60 min (n = 12, mean ± SD) (e), and lung virus titers (n = 6, mean ± SD) (f). Asterisk (*): below detection limit. Legends: filters are labelled as in Fig. 1b.To verify in vitro virus stability on the filters, an in vivo study was performed by infecting mice with virus incubated for 60 min on PP filters. As shown in Fig. 3e, Filterbare group exhibited 5% body weight loss at day 9 post-infection, reaching a body weight lower than that of salt-coated filter groups by 10–15%. Thus, significantly higher lung virus titers in the negative control group were observed in contrast to no detectable titers in the salt-coated filter groups (Fig. 3f).Strain-nonspecific virus deactivation and effects of storage under harsh environmental conditions on salt coating stabilityBroad-spectrum protection of salt-coated filters against multiple subtypes of viral aerosols was evaluated by investigating both lethal infectivity by penetrated virus in vivo and infectivity by virus collected on filters during filtration in vitro using A/Puerto Rico/08/1934 (PR/34 H1N1) and A/Vietnam/1203/2004 (VN/04 H5N1). Similarly to CA/09 H1N1, 100% of mice survived viral infection (PR/34 and VN/04), with no evidence of weight loss, due to higher filtration efficiency of salt-coated filter than that of bare filter (Fig. 4a). This is supported by no significant level of viral titer in the lung. In addition, as shown in Fig. 4b, salt-coated PP filters destroyed adsorbed influenza viruses irrespective of both subtypes and amount of coated salts.Figure 4: Strain- and environment-dependent performance of salt-coated filters.(a) Body weight change of mice infected with penetrated PR/34 H1N1 and VN/04 H5N1 viruses through Filterwet+600μL (n = 12, mean ± SD). (b) Virus titers of recovered viruses from bare and salt-coated filters (n = 4, mean ± SD; data for Filterwet, Filterwet+600μL and Filterwet+1200μL are overlapped). (c,d) Body weight change (c) and survival rate (d) of mice infected with dosage of penetrated virus through Filterwet+600μL before and after exposure to harsh environmental conditions (37 °C and 70% RH) for 1 day (filled square and open square overlap in (d)). (e) EDX mapping image of NaCl-coated Filterwet+600μL after incubation for 15 days at 37 °C and 70% RH (combination of Na (red) and Cl (green) mapping images). (f) XRD spectra of Filterwet+600μL before and after incubation at 37 °C 70% for 1 day and 15 days. Legends: filters are labelled as in Fig. 1b.The stability of salt coating on PP fibers was tested under harsh environmental conditions. Incubation at 37 °C and 70% relative humidity (RH) for 1 day did not cause any significant difference in filtration efficiency (t-test, P = 0.718) (Supplementary Fig. S8). As a result, all mice infected with dosage of penetrated virus through the filter stored at high temperature and RH displayed 100% survival with 7% of body weight loss (Fig. 4c,d). Even after 15 days of incubation, salts remained to coat PP fibers (Fig. 4e, and Supplementary Fig. S9a,b), despite change in grain orientation due to recrystallization (Fig. 4f, and Supplementary Fig. S10a,b).DiscussionDevelopment of a universally applicable, low-cost, and efficient mechanism for virus negation is regarded as a major challenge in public health against general airborne biological threats. This led us to propose a new concept of personal/public preventive and control measures using salt-recrystallization against pathogenic aerosols based on two hypotheses. The salt-coating can enhance adsorption of virus on the filter fibers and inactivate virus by the increase of osmotic pressure followed by the crystallization of salts. As shown in Fig. 2b, salt-coated filters exhibited significantly higher levels of filtration efficiency than bare filters. Notably, the bacterial filtration efficiency (BFE) reported by the mask manufacturer is 99%. The different value of filtration efficiency for bare filters obtained under our experimental conditions may be partially due to the use of aerosols with different biological origins. The FDA-recognized ASTM F2101 – 14 standard for evaluation of BFE exposes surgical masks to Staphylococcus aureus aerosols, by employing S. aureus ATCC 653834, which has an average diameter of about 1 μm. In this study, filtration efficiency was calculated following exposure of bare and salt-coated filters to influenza virus, which exhibits a smaller diameter than that of S. aureus by one order of magnitude. Additionally, whereas during BFE evaluation all three layers of surgical masks are used, in this work filtration efficiency refers to mask filters (middle layer). It is worth noting that the conditions for BFE standard evaluation (such as flow rate and time of application of flow) do not coincide with the experimental procedure we used for measurement of the filtration efficiency, which may further contribute to the different result. The enhanced filtration efficiency of salt-coated filters against influenza virus aerosols as compared to bare filters can be explained by the observed wetting of aerosols, favoring greater adhesion to salt-coated filters. Furthermore, the significant improvement in filtration efficiency resulted in complete protection of mice against lethal influenza aerosols, which demonstrates the high level of protection provided by salt-coated filters, outperforming currently used bare filters.Rapid loss of HA activity and viral infectivity on salt-coated filters can be explained by physical destruction of virus during recrystallization of coated salts. When the salt-coated filter is exposed to virus aerosols, salt crystals below the aerosol droplet dissolve to increase osmotic pressure to virus. Due to evaporation, the salt concentration of the droplet significantly increases and reaches the solubility limit, leading to recrystallization of salt. As a consequence, virus particles are exposed to increasing osmotic pressure during the drying process and are physically damaged by crystallization. As shown in Fig. 3e,f, the superior advantage of physically destroying the virus adsorbed to the salt-coated PP filters through natural salt crystallization process was further confirmed in vivo. According to previous reports, hyperosmotic stress (>541 mOsm) and crystallization induce membrane perturbation with irreversible deformation of the viral envelope and structural virus damage, respectively, resulting in infectivity loss of virus30,31. Therefore, our data support that the extensive level of infectivity loss associated with a salt recrystallization process caused by physical contact between virus aerosols and salt coating can be used in developing virus negation systems that are reusable without reprocessing.Similarly to CA/09 H1N1 aerosols, increased protection in vivo due to higher filtration efficiency of salt-coated filters compared to bare filters and deactivation of virus on salt-coated filters were observed following exposure to PR/34 H1N1 and VN/04 H5N1 (Fig. 4a,b). This suggests that salt-coated filters prevent virus penetration and destroy virus attached to the filter in a non-specific way. Furthermore, the performance of salt-coated filters was not degraded by storage at 37 °C and 70% RH, demonstrating that salt recrystallization-based filters can ensure protection even under harsh environmental conditions. Notably, for demonstration of the concept of salt-recrystallization based virus deactivation system, NaCl salt was used, which has a critical RH of 75% at 30 °C35. However, salts with higher critical RH can be easily used, such as ammonium sulfate, potassium chloride and potassium sulfate, which have critical RH of 80%, 84% and 96.3% at 30 °C, respectively35. This suggests that salt-coated filters may be developed for specific environmental conditions.In conclusion, we demonstrated that the developed salt-recrystallization based filtration system provides high filtration efficiency and successfully deactivates multiple subtypes of adsorbed viruses. Moreover, we have shown that stability of the salt coating is not compromised by high temperature and humidity, which suggests safe use and long-term storage/reuse at such environmental conditions. Although our tests are based on exposure to different types of influenza virus, the significance of these results for personal and public protective measures may be generally extended to enveloped respiratory viruses where infection and transmission can occur by aerosol. Our salt-coated filter unit can promise the development of long-term stable, versatile airborne pathogen negation system, without safety concerns. In fact, the destruction mechanism of viruses solely depends on the simple, yet robust naturally occurring salt recrystallization process, combining the destabilizing effects of salt crystal growth and concentration increase during drying of aerosols. This idea can be easily applied to a wide range of existing technologies to obtain low-cost, universal personal and public means of protection against airborne pathogens, such as masks and air filters in hospitals. Therefore, we believe that salt-recrystallization based virus deactivation system can contribute to global health by providing a more reliable means of preventing transmission and infection of pandemic or epidemic diseases and bioterrorism.MethodsBare and salt-coated filter samples preparationThe commercial surgical masks had a three-ply structure. The middle layer is the filter media, whereas the inner and outer layers provide support and protect the filter against wear and tear. The metal nose clips and elastic ear loops were removed and circular samples (radius: 3 cm) were cut from the masks. The PP filters (middle layer) were isolated by removing the inner and outer protective layers (bare filters, Filterbare). The coating solution was prepared by dissolving sodium chloride (NaCl; Sigma Aldrich, St. Louis, MO) in filtered DI water (0.22 μm pore size; Corning, Tewksbury, MA) under stirring at 400 rpm and 90 °C, followed by the addition of Tween 20 (Fisher Scientific) to a final concentration of 29.03 w/v% of NaCl and 1 v/v% of Tween 20. To obtain the salt-coated filters, the mask bare PP filters were pre-wet to contain approximately 600 μL of coating solution by incubating overnight at room temperature. Any remaining dry areas were removed by applying gentle strokes with tweezers to the filters while immersed in the coating solution. Subsequently, the filters were deposited in the desired volume of coating solution (0, 100, 300, 600, 900 and 1200 μL, of which corresponding membranes are abbreviated as Filterwet, Filterwet+100μL, Filterwet+300μL, Filterwet+600μL, Filterwet+900μL, and Filterwet+1200μL, respectively) on petri dishes (60 × 15 mm; Fisher Scientific) to control the amount of NaCl per unit area and dried in an oven (Isotemp Incubator, Fisher Scientific) at 37 °C for 1 day.Influenza virus preparationInfluenza viruses A/California/04/2009 (CA/09, H1N1), A/Puerto Rico/8/34 (PR/34, H1N1) and A/Vietnam/1203/2004 (VN/04, H5N1) were grown in 10-day old embryonated hen eggs, in which H5N1 virus was derived by reverse genetics from HPAI A/Vietnam/1203/200436. Influenza viruses were purified from allantoic fluid using discontinuous sucrose gradient (15%, 30% and 60%) layers following the previously reported procedure37.Aerosols exposure to filtersFor experiments involving aerosols exposure, an aerosol chamber (L × W × H = 145 × 145 × 150 mm; Emka Inc., Middletown, PA) was used (Fig. S11). It has a connection to the vacuum line and a circular aperture in the top wall (diameter: 22 mm) to exactly accommodate the cylindrical part (diameter: 20 mm, height; 20 mm) of the nebulizer unit that is below the aerosol generator (Aeroneb Lab Nebulizer System; Aerogen, Galway, Ireland). Bleach was used as trap between the chamber and the vacuum pump (Welch 2522C-10, 22 L/min; Niles, IL). The filters were placed on top of the chamber aperture and the nebulizer unit was inserted, ensuring the tight seal of the filters against the side of the aperture. 5 μL of virus stock were added to the nebulizer unit, aerosols (VMD 2.5–4 μm from manufacturer specifications) were generated for 30 sec and subsequently the desired vacuum level (3, 10 or 17 kPa) was applied, by manual control, three times in 1 sec cycles. Notably, in the case of bare filters, pressure was only applied for filtration efficiency tests.For all assays and analysis, suspensions of the filters were prepared as follows, unless otherwise indicated. To reconstitute virus adsorbed onto filters, virus-laden filters were immersed in 400 μL of sterilized DI water for about 5 min, and then removed after vortexing from the suspension. The virus suspension was centrifuged at 19,800 g and 4 °C for 10 min (Centrifuge 5810 R, Eppendorf, Hauppauge, NY), followed by resuspension of pellets in 70 μL of DI water to eliminate any interference from materials in supernatant during assays.Filtration efficiency testsThe filters were exposed to the virus aerosols at 3, 10 and 17 kPa and suspensions of the filters were obtained, as described above. The filtration efficiency was calculated as the ratio of the amount of virus (i.e., total proteins measured from the virus) reconstituted from the filter to that from the virus in the exposure aerosols. The concentration of virus in aerosols was determined by generating viral aerosols into a 15 mL centrifuge tube, containing 1 mL of DI water. After vortexing, virus concentrations (i.e., total protein concentration) were measured with bicinchoninic acid assay (BCA protein assay kit; Thermo Fischer scientific, Waltham, IL) with bovine serum albumin as a standard. In the case of virus reconstituted from salt-coated filters, virus-laden filter suspension was replaced with DI water prior to BCA assay.In vivo infection testsLethal infectivity of influenza viruses (CA/09 H1N1) was examined in 8 week old female inbred BALB/c mice (Nara Biotech; Seoul, Korea) by using the intranasal route. For bare and salt-coated filters, 12 mice per group were infected with individual penetration dosage of influenza virus through each filter. The penetration dosage of the virus through the filters (Filterbare, Filterwet, Filterwet+600μL, and Filterwet+1200μL) was calculated from the filtration efficiency at 10 kPa (near breathing pressure) using the relationship: penetration dosage = virus dosage in lethal aerosol × penetration efficiency (%)/100, where penetration efficiency (%) = 100 − filtration efficiency (%). To examine the effects of the aerosolization process on the viral infectivity change, two mice groups were infected with a lethal dose of virus before and after aerosol formation, which served as negative control groups. Body weight changes and survival rate of mice were monitored daily for 15 days. Mice with body weight loss greater than 25% were euthanized. All animal protocols were approved by the Kyung Hee University (KHU) Institutional Animal Care and Use Committee (IACUC). All animal experiments and husbandry involved in this work were conducted under the approved protocols and guidelines of KHU IACUC. KHU IACUC operates under National Veterinary Research and Quarantine Service (NVRQS), and animal welfare law and regulations of the WOAH-OIE (World organization for animal health).To test strain-dependent lethal infection behavior, mice (12 per group) were infected with the penetrated dosage of viral aerosols (PR/34 H1N1 and VN/04 H5N1 viruses) through Filterwet+600μL at 10 kPa. Time-dependent body weight change was monitored in the same manner described above.Lung viral titer and lung inflammatory cytokine assays after infectionOn day 4 after infection 6 mice of each group were sacrificed for the collection of lung samples. Lung virus titers were measured on six-well plates containing confluent MDCK cell monolayers. Inflammatory cytokines (IFN-γ) were determined using BD OptEIA mouse IFN-γ ELISA kit (BD Biosciences, San Jose, CA) following the manufacturer’s procedure.Test of viral infectivity change on filtersTo investigate the effects of salt-coating on viral infectivity loss, lethal influenza aerosols were exposed to four different types of filters (Filterbare, Filterwet, Filterwet+600μL, and Filterwet+1200μL). Since Filterbare exhibited almost complete penetration upon pressure application, aerosols were exposed to the bare filter in the absence of pressure and samples were carefully handled to prevent mechanical agitation. To measure time-dependent stability change of virus, virus-laden filters were incubated at ambient conditions for 0, 5, 15, and 60 min after aerosol exposure, and suspended in DI water to reconstitute virus at each time point. In vitro stability of virus was characterized by measuring hemagglutinin activity (HA) and virus titers at the same concentration as lethal dose30. The conformational stability of antigenic proteins was characterized by measuring intrinsic fluorescence using 0.1 mg/mL of virus suspension38. To investigate morphological change of virus, lipid stability of viral wall was characterized by nile red fluorescence (Sigma Aldrich), a fluorescent lipid stain, following manufacturer’s protocol39. A decrease in fluorescence intensity can be used to examine the level of disintegration of the virus. Both intrinsic and nile red fluorescence were measured by using a fluorimeter (LB 50B; PerkinElmer, Waltham, MA). Intensity changes of fluorescent spectra were compared relative to those of a control from virus stock.To test infectivity difference observed from in vitro findings, in vivo study was performed for the virus reconstituted from the filters (Filterbare, Filterwet, Filterwet+600μL, and Filterwet+1200μL) after incubation for 60 min at RT (aerosol exposure at 10 kPa, except for Filterbare). 12 mice per group were infected with a lethal dose of virus collected from each type of filter. Body weight change and lung virus titers were measured as described above.Effects of environmental conditions on the performance of salt-coated filterSalt-coated filters (Filterwet, Filterwet+600μL, and Filterwet+1200μL) were stored at 37 °C, 70% RH in an incubator (Maru Max; Rcom, Gyeonggi-do, South Korea) for 15 days. Every day, the filters were collected and incubated at ambient conditions for 5 min. At 1-day incubation, filtration efficiency was measured at 10 kPa from Filterwet+600μL, followed by in vivo infection test. Lethal infectivity between two different filter groups (before and after incubation at 37 °C, 70% RH) was compared by measuring body weight change and survival rate of mice after exposure to lethal CA/09 H1N1 aerosols. XRD analysis was performed to salt-coated filters incubated for 1 and 15 days, and SEM/EDX mapping analysis for 15-day incubated samples.Contact angle measurements and imaging of aerosolsThe bare and salt-coated filters were fixed with carbon tape (Ted Pella, Inc., Redding, CA) to a metal, flat substrate and 3 μL of DI water were added on the surface of the filters. The contact angles were measured from images collected with an optical microscope (10× lens, Motic SMZ-140; Motic, Richmond, Canada) at RT. Images of aerosols on filter fibers were obtained using a dispersive Raman microscope (Nicolet Almega XR; Fisher Scientific).Aerosol drying time on filtersThe bare and salt-coated filters were fixed with carbon tape to a metal, flat substrate and exposed to aerosols generated from 5 μL of Sulforhodamine B Dye solution (1 mM, Sigma-Aldrich). Aerosol drying time was determined with timer by observation with optical microscope.Electron microscopy analysisFor virus stability tests, bare and salt-coated filters were exposed to CA/09 H1N1 aerosols and, after 5 and 60 min incubation, virus was recovered by suspension of the filters, as described above. To study the effects of the coating formulation during aerosol drying independently from crystal growth, bare and salt-coated filters were immersed in DI water and removed after 60 min. Subsequently, virus was incubated in the obtained suspension for 60 min. Additionally, the virus suspension was centrifuged at 19,800 g and 4 °C for 10 min to collect the samples and suspend them in DI water. For TEM analysis (200 kV, JEOL JEM 2100; JEOL, Peabody, MA), samples were deposited on copper grid (Electron Microscopy Sciences, Hatfield, PA) and negatively stained with solution comprised of phosphotungstic acid hydrate (1.5 w/v%, pH = 7.0; Sigma-Aldrich, Oakville, Canada).To identify the morphology of salt-coated filters and recrystallized salts, SEM/EDX analysis was performed for bare and salt-coated filters after coating with 10 nm thick gold layer. Scanning electron microscopy analysis (Hitachi S-3000N; Hitachi, Toronto, Canada) was operated in secondary electron mode at 20 kV and EDX analysis was obtained with EDX detector (Oxford Instruments, Concord, MA).XRD analysisTo confirm the formation of crystalline NaCl coating during drying process and its stability during storage at 37 °C and 70% RH, XRD analysis (BRU-1098; Bruker, Billerica, MA) was performed at different coating conditions. Filters (1 × 1 cm) were mounted on a slide glass for XRD analysis (θ–2θ mode) using a CuKα radiation.Statistical analysisTo compare multiple conditions, Student’s t-test, One-way analysis of variance (ANOVA), and general linear model were used (Minitab release 14; Minitab, State College, PA). P value of less than 0.05 was considered to be significant.Additional InformationHow to cite this article: Quan, F.-S. et al. Universal and reusable virus deactivation system for respiratory protection. Sci. Rep. 7, 39956; doi: 10.1038/srep39956 (2017).Publisher's note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.\n \n\n References1Tellier, R. Review of aerosol transmission of influenza A virus. Emerging Infect. Dis. 12, 1657–1662 (2006).2Weber, T. P. & Stilianakis, N. I. Inactivation of influenza A viruses in the environment and modes of transmission: a critical review. J. Infect. 57, 361–373 (2008).3Duy, J. et al. Circulating microRNA profiles of Ebola virus infection. Sci. Rep. 6, 24496 (2016).4Morse, S. S., Garwin, R. L. & Olsiewski, P. J. Next flu pandemic: what to do until the vaccine arrives? Science 314, 929–929 (2006).5CDC. Laboratory performance evaluation of N95 filtering facepiece respirators, 1996. MMWR, Morbidity and Mortality Weekly Report 47, 1045 (1998).6Seto, W. H. et al. Effectiveness of precautions against droplets and contact in prevention of nosocomial transmission of severe acute respiratory syndrome (SARS). Lancet 361, 1519–1520 (2003).7Bałazy, A. et al. Do N95 respirators provide 95% protection level against airborne viruses, and how adequate are surgical masks? Am. J. Infect. Control 34, 51–57 (2006).8Loeb, M. et al. Surgical mask vs N95 respirator for preventing influenza among health care workers: a randomized trial. JAMA 302, 1865–1871 (2009).9Bunyan, D., Ritchie, L., Jenkins, D. & Coia, J. E. Respiratory and facial protection: a critical review of recent literature. J. Hosp. Infect. 85, 165–169 (2013).10Zhu, C., Lin, C. H. & Cheung, C. S. Inertial impaction-dominated fibrous filtration with rectangular or cylindrical fibers. Powder Technol. 112, 149–162 (2000).11Lee, K. W. & Liu, B. Y. H. Theoretical study of aerosol filtration by fibrous filters. Aerosol Sci. Tech. 1, 147–161 (1982).12Duarte Fo, O. B., Marra, W. D., Kachan, G. C. & Coury, J. R. Filtration of electrified solid particles. Ind. Eng. Chem. Res 39, 3884–3895 (2000).13Kraemer, H. F. & Johnstone, H. F. Collection of aerosol particles in presence of electrostatic fields. Ind. Eng. Chem. 47, 2426–2434 (1955).14Ranz, W. E. & Wong, J. B. Impaction of dust and smoke particles on surface and body collectors. Ind. Eng. Chem. 44, 1371–1381 (1952).15Siegel, J. D., Rhinehart, E., Jackson, M. & Chiarello, L. 2007 guideline for isolation precautions: preventing transmission of infectious agents in health care settings. Am. J. Infect. Control 35, S65–S164 (2007).16Coia, J. E. et al. Guidance on the use of respiratory and facial protection equipment. J. Hosp. Infect. 85, 170–182 (2013).17Ratnesar-Shumate, S. et al. Evaluation of physical capture efficiency and disinfection capability of an iodinated biocidal filter medium. Aerosol Air Qual. Res. 8, 1–18 (2008).18Lee, J. H., Wu, C. Y., Wysocki, K. M. & Farrah, S. & Wander, J. Efficacy of iodine-treated biocidal filter media against bacterial spore aerosols. J. Appl. Microbiol. 105, 1318–1326 (2008).19Badrossamay, M. R. & Sun, G. Acyclic halamine polypropylene polymer: effect of monomer structure on grafting efficiency, stability and biocidal activities. React. Funct. Polym. 68, 1636–1645 (2008).20Zhao, N. & Liu, S. Thermoplastic semi-IPN of polypropylene (PP) and polymeric N-halamine for efficient and durable antibacterial activity. Eur. Polym. J. 47, 1654–1663 (2011).21Cerkez, I., Worley, S. D., Broughton, R. M. & Huang, T. S. Antimicrobial surface coatings for polypropylene nonwoven fabrics. React. Funct. Polym. 73, 1412–1419 (2013).22Davison, A. M. Pathogen inactivation and filtration efficacy of a new anti-microbial and anti-viral surgical facemask and N95 against dentistry-associated microorganisms. International dentistry Australasian edition 7, 36–42 (2012).23Borkow, G., Zhou, S. S., Page, T. & Gabbay, J. A novel anti-influenza copper oxide containing respiratory face mask. PLoS One 5, e11295 (2010).24Borkow, G. & Gabbay, J. Putting copper into action: copper-impregnated products with potent biocidal activities. FASEB J. 18, 1728–1730 (2004).25Li, Y., Leung, P., Yao, L., Song, Q. W. & Newton, E. Antimicrobial effect of surgical masks coated with nanoparticles. J. Hosp. Infect. 62, 58–63 (2006).26Lee, J. H. et al. Assessment of iodine-treated filter media for removal and inactivation of MS2 bacteriophage aerosols. J. Appl. Microbiol. 107, 1912–1923 (2009).27Lore, M. B. et al. Performance of conventional and antimicrobial-treated filtering facepiece respirators challenged with biological aerosols. J. Occup. Environ. Hyg. 9, 69–80 (2012).28Rengasamy, S., Fisher, E. & Shaffer, R. E. Evaluation of the survivability of MS2 viral aerosols deposited on filtering face piece respirator samples incorporating antimicrobial technologies. Am. J. Infect. Control 38, 9–17 (2010).29Windisch, W., Hennings, E., Sorichter, S., Hamm, H. & Criee, C. P. Peak or plateau maximal inspiratory mouth pressure: which is best? Eur. Respir. J. 23, 708–713 (2004).30Choi, H.-J. et al. Stability of influenza vaccine coated onto microneedles. Biomaterials 33, 3756–3769 (2012).31Choi, H.-J. et al. Stability of whole inactivated influenza virus vaccine during coating onto metal microneedles. J. Control. Release 166, 159–171 (2013).32Baldwin, R. L. How hofmeister ion interactions affect protein stability. Biophys. J. 71, 2056–2063 (1996).33Choi, H.-J. et al. Effect of osmotic pressure on the stability of whole inactivated influenza vaccine for coating on microneedles. PLoS one 10, e0134431 (2015).34ASTM International. ASTM F2101 - 14 Standard Test Method for Evaluating the Bacterial Filtration Efficiency (BFE) of Medical Face Mask Materials, Using a Biological Aerosol of Staphylococcus aureus (2014).35Adams, J. R. & Merz, A. R. Hygroscopicity of fertilizer materials and mixtures. Ind. Eng. Chem. Res. 21, 305–307 (1929).36Song, J.-M. et al. Protective immunity against H5N1 influenza virus by a single dose vaccination with virus-like particles. Virology 405, 165–175 (2010).37Quan, F. S. et al. A bivalent influenza VLP vaccine confers complete inhibition of virus replication in lungs. Vaccine 26, 3352–3361 (2008).38Choi, H.-J., Ebersbacher, C. F., Kim, M., Kang, S. & Montemagno, C. D. A mechanistic study on the destabilization of whole inactivated influenza virus vaccine in gastric environment. PLoS one 8, e66316 (2013).39Greenspan, P., Mayer, E. P. & Fowler, S. D. Nile red: a selective fluorescent stain for intracellular lipid droplets. J. Cell Biol. 100, 965–973 (1985).Download referencesAcknowledgementsThis research was financially supported by startup funds from University of Alberta (H.J.C.), and grants from National Research Foundation of Korea (NRF) (NRF-2014R1A2A2A01004899) and Ministry of Health & Welfare, Republic of Korea (HI15C2928).Author informationAuthor notesFu-Shi Quan and Ilaria Rubino: These authors contributed equally to this work.AffiliationsDepartment of Medical Zoology, Kyung Hee University School of Medicine, Seoul, 130-701, KoreaFu-Shi QuanDepartment of Chemical and Materials Engineering, University of Alberta, Edmonton, AB T6G 1H9, CanadaIlaria Rubino, Brendan Koch & Hyo-Jick ChoiDepartment of Biomedical Science, Graduate School, Kyung Hee University, Seoul, 130-701, KoreaSu-Hwa LeeAuthorsSearch for Fu-Shi Quan in:Search for Ilaria Rubino in:Search for Su-Hwa Lee in:Search for Brendan Koch in:Search for Hyo-Jick Choi in:ContributionsH.J.C. conceived and designed the experiments. F.S.Q., I.R., S.H.L., B.K., and H.J.C. performed the experiments. F.S.Q., I.R., S.H.L., B.K., and H.J.C. analyzed the data. I.R. and H.J.C. wrote the manuscript. F.S.Q. and B.K. edited the manuscript.Corresponding authorCorrespondence to\n Hyo-Jick Choi.Ethics declarations\n \n Competing interests\n The authors declare no competing financial interests.\n \n Supplementary informationRights and permissionsThis work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/Reprints and PermissionsAbout this articleCite this articleQuan, F., Rubino, I., Lee, S. et al. Universal and reusable virus deactivation system for respiratory protection.\n Sci Rep 7, 39956 (2017). https://doi.org/10.1038/srep39956Download citationReceived: 04 August 2016Accepted: 30 November 2016Published: 04 January 2017DOI: https://doi.org/10.1038/srep39956\n\n \n \n \n Further reading\n \n \n \n \n \n \n Survival of Microorganisms on Nonwovens Used for the Construction of Filtering Facepiece Respirators\n \n \n \n \n Katarzyna Majchrzycka, Małgorzata Okrasa, Justyna Szulc, Anita Jachowicz[…] & Beata Gutarowska- Show fewer authors\n \n \n International Journal of Environmental Research and Public Health\n (2019)\n \n \n \n \n \n \n Respiratory Protection against Pandemic and Epidemic Diseases\n \n \n \n \n Ilaria Rubino & Hyo-Jick Choi\n \n \n Trends in Biotechnology\n (2017)\n \n \n \n \n \n \n \n \n\n \n CommentsBy submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate."],["topImageUrl",null],["error",null]]}]}],["text","WIKILINK\n最新科技資訊\n\n \n口罩能否反復使用？浸鹽水後可100%讓H1N1病毒滅活，但對其他病毒需謹慎 | 《科學報告》期刊\nFebruary 5, 2020 admin\n曉查發自凹非寺 \n量子位報道 | 公眾號 QbitAI\n一場新冠肺炎讓中國的口罩嚴重告急。\n\n每齣一次門就要廢掉一個口罩，傢裏的口罩即將用完，藥店和網上都買不到，真是急死人瞭。\n\n彆怕，早在2016年，來自韓國慶熙大學的研究者就發現，口罩隻需用普通食鹽水處理後，就具有瞭一定的殺滅病毒能力，至少對2009年爆發的H1N1流感病毒有效。\n\n他們將處理後的口罩來過濾充滿H1N1病毒的空氣，然後再給小白鼠呼吸，結果發現小鼠的存活率達到瞭100%。\n\n\n\n\n \n空氣中的病毒濃度達到瞭緻死量。作為對照，如果使用普通醫用口罩過濾，小白鼠在4天後就會齣現嚴重的肺炎，10天後全部死亡。\n\n這篇文章發錶在2017年1月Nature旗下的Scientific Reports期刊上。\n\n\n\n△實驗中用到的過濾裝置\n早在3年前，該文章就預測瞭口罩短缺的可能性。\n\n作者在開頭說，在疫病大流行時期，生産能力不足和製造過程耗時，會導緻口罩供應有限。\n\nN95口罩雖然可以過濾空氣中形成的病毒氣溶膠，但是在嚴重的疫情麵前，大規模使用N95口罩是不切實際的，而且價格昂貴。\n\n過去的SARS、H1N1、MERS都錶明，醫用外科口罩是被公眾最廣泛采用的個人防護措施，盡管人們對它的效果存在爭議。\n\n因此需要開發一種通用、低成本且有效的防護病毒機製，最好能用在普通醫用口罩上。\n\n\n\n\n\n\n研究人員配置瞭濃度29.03 w/v%的氯化鈉溶液，並在其中加入瞭一些Tween 20錶麵活性劑，將口罩浸潤，然後在37℃的環境下乾燥一天。\n\n文章指齣，經過這種方法處理後的口罩無需再次處理即可重復使用。最終實驗發現，塗有氯化鈉的口罩上的病毒在5分鍾後就失去瞭活性。\n\n\n\n\n\n\n從顯微鏡分析來看，病毒氣溶膠乾燥時間約為3分鍾，錶明在5分鍾時觀察到的病毒破壞與乾燥引起的鹽結晶有關。\n\n作者推測，可能是帶有水分的病毒被吸附在口罩錶麵，濕潤後又重結晶的鹽破壞瞭病毒的結構。\n\n既然對H1N1病毒，自然就有網友很好奇，這種處理方法是否也對新型冠狀病毒有效。\n\n一些網友對此提齣瞭質疑，認為鹽結晶可能減小瞭口罩的通氣量，而且實驗隻對H1N1病毒進行瞭驗證，是否實用還有待商榷。\n\n所以在未經實驗驗證的情況下，請不要用這種方法來過濾新型冠狀病毒。口罩還是要保證一用一換，切不可自行處理後就重復使用，否則仍有被感染的風險。\n\n論文地址：\nhttps://www.nature.com/articles/srep39956#Sec8\n\n— 完 —\n\n"],["id","obwrpyiajxnx"],["createdAt","2020-02-09T01:53:22.130Z"]]},{"__iterable":"Map","data":[["replyCount",1],["references",{"__iterable":"List","data":[{"__iterable":"Map","data":[["type","LINE"]]}]}],["replyRequestCount",1],["user",{"__iterable":"Map","data":[["id","j4S8C_n1X_AVJRpVDJaAWXXARts8Z96DWjC7LTqy8kpttWQvw"],["name","☞健談的坪林韋納爾☜"],["avatarUrl",null],["level",0]]}],["hyperlinks",{"__iterable":"List","data":[{"__iterable":"Map","data":[["title","Clinical features of patients infected with 2019 novel coronavirus in Wuhan, China"],["url","https://www.thelancet.com/journals/lancet/article/PIIS0140-6736(20)30183-5/fulltext"],["summary","SummaryBackgroundA recent cluster of pneumonia cases in Wuhan, China, was caused by a novel betacoronavirus, the 2019 novel coronavirus (2019-nCoV). We report the epidemiological, clinical, laboratory, and radiological characteristics and treatment and clinical outcomes of these patients.MethodsAll patients with suspected 2019-nCoV were admitted to a designated hospital in Wuhan. We prospectively collected and analysed data on patients with laboratory-confirmed 2019-nCoV infection by real-time RT-PCR and next-generation sequencing. Data were obtained with standardised data collection forms shared by the International Severe Acute Respiratory and Emerging Infection Consortium from electronic medical records. Researchers also directly communicated with patients or their families to ascertain epidemiological and symptom data. Outcomes were also compared between patients who had been admitted to the intensive care unit (ICU) and those who had not.FindingsBy Jan 2, 2020, 41 admitted hospital patients had been identified as having laboratory-confirmed 2019-nCoV infection. Most of the infected patients were men (30 [73%] of 41); less than half had underlying diseases (13 [32%]), including diabetes (eight [20%]), hypertension (six [15%]), and cardiovascular disease (six [15%]). Median age was 49·0 years (IQR 41·0–58·0). 27 (66%) of 41 patients had been exposed to Huanan seafood market. One family cluster was found. Common symptoms at onset of illness were fever (40 [98%] of 41 patients), cough (31 [76%]), and myalgia or fatigue (18 [44%]); less common symptoms were sputum production (11 [28%] of 39), headache (three [8%] of 38), haemoptysis (two [5%] of 39), and diarrhoea (one [3%] of 38). Dyspnoea developed in 22 (55%) of 40 patients (median time from illness onset to dyspnoea 8·0 days [IQR 5·0–13·0]). 26 (63%) of 41 patients had lymphopenia. All 41 patients had pneumonia with abnormal findings on chest CT. Complications included acute respiratory distress syndrome (12 [29%]), RNAaemia (six [15%]), acute cardiac injury (five [12%]) and secondary infection (four [10%]). 13 (32%) patients were admitted to an ICU and six (15%) died. Compared with non-ICU patients, ICU patients had higher plasma levels of IL2, IL7, IL10, GSCF, IP10, MCP1, MIP1A, and TNFα.InterpretationThe 2019-nCoV infection caused clusters of severe respiratory illness similar to severe acute respiratory syndrome coronavirus and was associated with ICU admission and high mortality. Major gaps in our knowledge of the origin, epidemiology, duration of human transmission, and clinical spectrum of disease need fulfilment by future studies.FundingMinistry of Science and Technology, Chinese Academy of Medical Sciences, National Natural Science Foundation of China, and Beijing Municipal Science and Technology Commission.IntroductionCoronaviruses are enveloped non-segmented positive-sense RNA viruses belonging to the family Coronaviridae and the order Nidovirales and broadly distributed in humans and other mammals.1Richman DD Whitley RJ Hayden FG Although most human coronavirus infections are mild, the epidemics of the two betacoronaviruses, severe acute respiratory syndrome coronavirus (SARS-CoV)2Ksiazek TG Erdman D Goldsmith CS et al.A novel coronavirus associated with severe acute respiratory syndrome., 3Kuiken T Fouchier RAM Schutten M et al.Newly discovered coronavirus as the primary cause of severe acute respiratory syndrome., 4Drosten C Günther S Preiser W et al.Identification of a novel coronavirus in patients with severe acute respiratory syndrome. and Middle East respiratory syndrome coronavirus (MERS-CoV),5de Groot RJ Baker SC Baric RS et al.Middle East respiratory syndrome coronavirus (MERS-CoV): announcement of the Coronavirus Study Group., 6Zaki AM van Boheemen S Bestebroer TM Osterhaus ADME Fouchier RAM Isolation of a novel coronavirus from a man with pneumonia in Saudi Arabia. have caused more than 10 000 cumulative cases in the past two decades, with mortality rates of 10% for SARS-CoV and 37% for MERS-CoV.7WHOSummary of probable SARS cases with onset of illness from 1 November 2002 to 31 July 2003., 8WHOMiddle East respiratory syndrome coronavirus (MERS-CoV). The coronaviruses already identified might only be the tip of the iceberg, with potentially more novel and severe zoonotic events to be revealed.In December, 2019, a series of pneumonia cases of unknown cause emerged in Wuhan, Hubei, China, with clinical presentations greatly resembling viral pneumonia.9WHONovel coronavirus – China. Deep sequencing analysis from lower respiratory tract samples indicated a novel coronavirus, which was named 2019 novel coronavirus (2019-nCoV). Thus far, more than 800 confirmed cases, including in health-care workers, have been identified in Wuhan, and several exported cases have been confirmed in other provinces in China, and in Thailand, Japan, South Korea, and the USA.10WHONovel coronavirus – Thailand (ex-China)., 11WHONovel coronavirus – Japan (ex-China)., 12WHONovel coronavirus – Republic of Korea (ex-China)., 13CDCFirst travel-related case of 2019 novel coronavirus detected in United States.We aim to describe epidemiological, clinical, laboratory, and radiological characteristics, treatment, and outcomes of patients confirmed to have 2019-nCoV infection, and to compare the clinical features between intensive care unit (ICU) and non-ICU patients. We hope our study findings will inform the global community of the emergence of this novel coronavirus and its clinical features.Methods PatientsFollowing the pneumonia cases of unknown cause reported in Wuhan and considering the shared history of exposure to Huanan seafood market across the patients, an epidemiological alert was released by the local health authority on Dec 31, 2019, and the market was shut down on Jan 1, 2020. Meanwhile, 59 suspected cases with fever and dry cough were transferred to a designated hospital starting from Dec 31, 2019. An expert team of physicians, epidemiologists, virologists, and government officials was soon formed after the alert.Since the cause was unknown at the onset of these emerging infections, the diagnosis of pneumonia of unknown cause in Wuhan was based on clinical characteristics, chest imaging, and the ruling out of common bacterial and viral pathogens that cause pneumonia. Suspected patients were isolated using airborne precautions in the designated hospital, Jin Yin-tan Hospital (Wuhan, China), and fit-tested N95 masks and airborne precautions for aerosol-generating procedures were taken. This study was approved by the National Health Commission of China and Ethics Commission of Jin Yin-tan Hospital (KY-2020-01.01). Written informed consent was waived by the Ethics Commission of the designated hospital for emerging infectious diseases. ProceduresLocal centres for disease control and prevention collected respiratory, blood, and faeces specimens, then shipped them to designated authoritative laboratories to detect the pathogen (NHC Key Laboratory of Systems Biology of Pathogens and Christophe Mérieux Laboratory, Beijing, China). A novel coronavirus, which was named 2019-nCoV, was isolated then from lower respiratory tract specimen and a diagnostic test for this virus was developed soon after that.14Tan W Zhao X Ma X et al.A novel coronavirus genome identified in a cluster of pneumonia cases — Wuhan, China 2019−2020. Of 59 suspected cases, 41 patients were confirmed to be infected with 2019-nCoV. The presence of 2019-nCoV in respiratory specimens was detected by next-generation sequencing or real-time RT-PCR methods. The primers and probe target to envelope gene of CoV were used and the sequences were as follows: forward primer 5′-TCAGAATGCCAATCTCCCCAAC-3′; reverse primer 5′-AAAGGTCCACCCGATACATTGA-3′; and the probe 5′CY5-CTAGTTACACTAGCCATCCTTACTGC-3′BHQ1. Conditions for the amplifications were 50°C for 15 min, 95°C for 3 min, followed by 45 cycles of 95°C for 15 s and 60°C for 30 s.Initial investigations included a complete blood count, coagulation profile, and serum biochemical test (including renal and liver function, creatine kinase, lactate dehydrogenase, and electrolytes). Respiratory specimens, including nasal and pharyngeal swabs, bronchoalveolar lavage fluid, sputum, or bronchial aspirates were tested for common viruses, including influenza, avian influenza, respiratory syncytial virus, adenovirus, parainfluenza virus, SARS-CoV and MERS-CoV using real-time RT-PCR assays approved by the China Food and Drug Administration. Routine bacterial and fungal examinations were also performed.Given the emergence of the 2019-nCoV pneumonia cases during the influenza season, antibiotics (oral and intravenous) and oseltamivir (orally 75 mg twice daily) were empirically administered. Corticosteroid therapy (methylprednisolone 40–120 mg per day) was given as a combined regimen if severe community-acquired pneumonia was diagnosed by physicians at the designated hospital. Oxygen support (eg, nasal cannula and invasive mechanical ventilation) was administered to patients according to the severity of hypoxaemia. Repeated tests for 2019-nCoV were done in patients confirmed to have 2019-nCoV infection to show viral clearance before hospital discharge or discontinuation of isolation. Data collectionWe reviewed clinical charts, nursing records, laboratory findings, and chest x-rays for all patients with laboratory-confirmed 2019-nCoV infection who were reported by the local health authority. The admission data of these patients was from Dec 16, 2019, to Jan 2, 2020. Epidemiological, clinical, laboratory, and radiological characteristics and treatment and outcomes data were obtained with standardised data collection forms (modified case record form for severe acute respiratory infection clinical characterisation shared by the International Severe Acute Respiratory and Emerging Infection Consortium) from electronic medical records. Two researchers also independently reviewed the data collection forms to double check the data collected. To ascertain the epidemiological and symptom data, which were not available from electronic medical records, the researchers also directly communicated with patients or their families to ascertain epidemiological and symptom data. Cytokine and chemokine measurementTo characterise the effect of coronavirus on the production of cytokines or chemokines in the acute phase of the illness, plasma cytokines and chemokines (IL1B, IL1RA, IL2, IL4, IL5, IL6, IL7, IL8 (also known as CXCL8), IL9, IL10, IL12p70, IL13, IL15, IL17A, Eotaxin (also known as CCL11), basic FGF2, GCSF (CSF3), GMCSF (CSF2), IFNγ, IP10 (CXCL10), MCP1 (CCL2), MIP1A (CCL3), MIP1B (CCL4), PDGFB, RANTES (CCL5), TNFα, and VEGFA were measured using Human Cytokine Standard 27-Plex Assays panel and the Bio-Plex 200 system (Bio-Rad, Hercules, CA, USA) for all patients according to the manufacturer's instructions. The plasma samples from four healthy adults were used as controls for cross-comparison. The median time from being transferred to a designated hospital to the blood sample collection was 4 days (IQR 2–5). Detection of coronavirus in plasmaEach 80 μL plasma sample from the patients and contacts was added into 240 μL of Trizol LS (10296028; Thermo Fisher Scientific, Carlsbad, CA, USA) in the Biosafety Level 3 laboratory. Total RNA was extracted by Direct-zol RNA Miniprep kit (R2050; Zymo research, Irvine, CA, USA) according to the manufacturer's instructions and 50 μL elution was obtained for each sample. 5 μL RNA was used for real-time RT-PCR, which targeted the NP gene using AgPath-ID One-Step RT-PCR Reagent (AM1005; Thermo Fisher Scientific). The final reaction mix concentration of the primers was 500 nM and probe was 200 nM. Real-time RT-PCR was performed using the following conditions: 50°C for 15 min and 95°C for 3 min, 50 cycles of amplification at 95°C for 10 s and 60°C for 45 s. Since we did not perform tests for detecting infectious virus in blood, we avoided the term viraemia and used RNAaemia instead. RNAaemia was defined as a positive result for real-time RT-PCR in the plasma sample. DefinitionsAcute respiratory distress syndrome (ARDS) and shock were defined according to the interim guidance of WHO for novel coronavirus.9WHONovel coronavirus – China. Hypoxaemia was defined as arterial oxygen tension (PaO2) over inspiratory oxygen fraction (FIO2) of less than 300 mm Hg.15Sanz F Gimeno C Lloret T et al.Relationship between the presence of hypoxemia and the inflammatory response measured by C-reactive protein in bacteremic pneumococcal pneumonia. Acute kidney injury was identified and classified on the basis of the highest serum creatinine level or urine output criteria according to the kidney disease improving global outcomes classification.16Kidney disease: improving global outcomes (KDIGO) acute kidney injury work groupKDIGO clinical practice guideline for acute kidney injury. Secondary infection was diagnosed if the patients had clinical symptoms or signs of nosocomial pneumonia or bacteraemia, and was combined with a positive culture of a new pathogen from a lower respiratory tract specimen (including the sputum, transtracheal aspirates, or bronchoalveolar lavage fluid, or from blood samples taken ≥48 h after admission).17Garner JS Jarvis WR Emori TG Horan TC Hughes JM CDC definitions for nosocomial infections, 1988. Cardiac injury followed the definition used in our previous study in H7N9 patients.18Gao C Wang Y Gu X et al.Association between cardiac injury and mortality in hospitalized patients infected with avian influenza A (H7N9) virus. In brief, cardiac injury was diagnosed if serum levels of cardiac biomarkers (eg, troponin I) were above the 99th percentile upper reference limit, or new abnormalities were shown in electrocardiography and echocardiography. Statistical analysisContinuous variables were expressed as median (IQR) and compared with the Mann-Whitney U test; categorical variables were expressed as number (%) and compared by χ2 test or Fisher's exact test between ICU care and no ICU care groups. Boxplots were drawn to describe plasma cytokine and chemokine concentrations.A two-sided α of less than 0·05 was considered statistically significant. Statistical analyses were done using the SAS software, version 9.4, unless otherwise indicated. Role of the funding sourceThe funder of the study had no role in study design, data collection, data analysis, data interpretation, or writing of the report. The corresponding authors had full access to all the data in the study and had final responsibility for the decision to submit for publication.ResultsBy Jan 2, 2020, 41 admitted hospital patients were identified as laboratory-confirmed 2019-nCoV infection in Wuhan. 20 [49%]) of the 2019-nCoV-infected patients were aged 25–49 years, and 14 (34%) were aged 50–64 years (figure 1A). The median age of the patients was 49·0 years (IQR 41·0–58·0; table 1). In our cohort of the first 41 patients as of Jan 2, no children or adolescents were infected. Of the 41 patients, 13 (32%) were admitted to the ICU because they required high-flow nasal cannula or higher-level oxygen support measures to correct hypoxaemia. Most of the infected patients were men (30 [73%]); less than half had underlying diseases (13 [32%]), including diabetes (eight [20%]), hypertension (six [15%]), and cardiovascular disease (six [15%]).Table 1Demographics and baseline characteristics of patients infected with 2019-nCoVData are median (IQR), n (%), or n/N (%), where N is the total number of patients with available data. p values comparing ICU care and no ICU care are from χ2 test, Fisher's exact test, or Mann-Whitney U test. 2019-nCoV=2019 novel coronavirus. ICU=intensive care unit.\n Open table in a new tab\n 27 (66%) patients had direct exposure to Huanan seafood market (figure 1B). Market exposure was similar between the patients with ICU care (nine [69%]) and those with non-ICU care (18 [64%]). The symptom onset date of the first patient identified was Dec 1, 2019. None of his family members developed fever or any respiratory symptoms. No epidemiological link was found between the first patient and later cases. The first fatal case, who had continuous exposure to the market, was admitted to hospital because of a 7-day history of fever, cough, and dyspnoea. 5 days after illness onset, his wife, a 53-year-old woman who had no known history of exposure to the market, also presented with pneumonia and was hospitalised in the isolation ward.The most common symptoms at onset of illness were fever (40 [98%] of 41 patients), cough (31 [76%]), and myalgia or fatigue (18 [44%]); less common symptoms were sputum production (11 [28%] of 39), headache (three [8%] of 38), haemoptysis (two [5%] of 39), and diarrhoea (one [3%] of 38; table 1). More than half of patients (22 [55%] of 40) developed dyspnoea. The median duration from illness onset to dyspnoea was 8·0 days (IQR 5·0–13·0). The median time from onset of symptoms to first hospital admission was 7·0 days (4·0–8·0), to shortness of breath was 8·0 days (5·0–13·0), to ARDS was 9·0 days (8·0–14·0), to mechanical ventilation was 10·5 days (7·0–14·0), and to ICU admission was 10·5 days (8·0–17·0; figure 2).The blood counts of patients on admission showed leucopenia (white blood cell count less than 4 × 109/L; ten [25%] of 40 patients) and lymphopenia (lymphocyte count <1·0 × 109/L; 26 [63%] patients; table 2). Prothrombin time and D-dimer level on admission were higher in ICU patients (median prothrombin time 12·2 s [IQR 11·2–13·4]; median D-dimer level 2·4 mg/L [0·6–14·4]) than non-ICU patients (median prothrombin time 10·7 s [9·8–12·1], p=0·012; median D-dimer level 0·5 mg/L [0·3–0·8], p=0·0042). Levels of aspartate aminotransferase were increased in 15 (37%) of 41 patients, including eight (62%) of 13 ICU patients and seven (25%) of 28 non-ICU patients. Hypersensitive troponin I (hs-cTnI) was increased substantially in five patients, in whom the diagnosis of virus-related cardiac injury was made.Table 2Laboratory findings of patients infected with 2019-nCoV on admission to hospitalData are median (IQR) or n/N (%), where N is the total number of patients with available data. p values comparing ICU care and no ICU care are from χ2, Fisher's exact test, or Mann-Whitney U test. 2019-nCoV=2019 novel coronavirus. ICU=intensive care unit.\n Open table in a new tab\n Most patients had normal serum levels of procalcitonin on admission (procalcitonin <0·1 ng/mL; 27 [69%] patients; table 2). Four ICU patients developed secondary infections. Three of the four patients with secondary infection had procalcitonin greater than 0·5 ng/mL (0·69 ng/mL, 1·46 ng/mL, and 6·48 ng/mL).On admission, abnormalities in chest CT images were detected among all patients. Of the 41 patients, 40 (98%) had bilateral involvement (table 2). The typical findings of chest CT images of ICU patients on admission were bilateral multiple lobular and subsegmental areas of consolidation (figure 3A). The representative chest CT findings of non-ICU patients showed bilateral ground-glass opacity and subsegmental areas of consolidation (figure 3B). Later chest CT images showed bilateral ground-glass opacity, whereas the consolidation had been resolved (figure 3C).Initial plasma IL1B, IL1RA, IL7, IL8, IL9, IL10, basic FGF, GCSF, GMCSF, IFNγ, IP10, MCP1, MIP1A, MIP1B, PDGF, TNFα, and VEGF concentrations were higher in both ICU patients and non-ICU patients than in healthy adults (appendix pp 6–7). Plasma levels of IL5, IL12p70, IL15, Eotaxin, and RANTES were similar between healthy adults and patients infected with 2019-nCoV. Further comparison between ICU and non-ICU patients showed that plasma concentrations of IL2, IL7, IL10, GCSF, IP10, MCP1, MIP1A, and TNFα were higher in ICU patients than non-ICU patients.All patients had pneumonia. Common complications included ARDS (12 [29%] of 41 patients), followed by RNAaemia (six [15%] patients), acute cardiac injury (five [12%] patients), and secondary infection (four [10%] patients; table 3). Invasive mechanical ventilation was required in four (10%) patients, with two of them (5%) had refractory hypoxaemia and received extracorporeal membrane oxygenation as salvage therapy. All patients were administered with empirical antibiotic treatment, and 38 (93%) patients received antiviral therapy (oseltamivir). Additionally, nine (22%) patients were given systematic corticosteroids. A comparison of clinical features between patients who received and did not receive systematic corticosteroids is in the appendix (pp 1–5).Table 3Treatments and outcomes of patients infected with 2019-nCoVData are median (IQR) or n (%). p values are comparing ICU care and no ICU care. 2019-nCoV=2019 novel coronavirus. ICU=intensive care unit. NA=not applicable. ECMO=extracorporeal membrane oxygenation.\n Open table in a new tab\n As of Jan 22, 2020, 28 (68%) of 41 patients have been discharged and six (15%) patients have died. Fitness for discharge was based on abatement of fever for at least 10 days, with improvement of chest radiographic evidence and viral clearance in respiratory samples from upper respiratory tract.DiscussionWe report here a cohort of 41 patients with laboratory-confirmed 2019-nCoV infection. Patients had serious, sometimes fatal, pneumonia and were admitted to the designated hospital in Wuhan, China, by Jan 2, 2020. Clinical presentations greatly resemble SARS-CoV. Patients with severe illness developed ARDS and required ICU admission and oxygen therapy. The time between hospital admission and ARDS was as short as 2 days. At this stage, the mortality rate is high for 2019-nCoV, because six (15%) of 41 patients in this cohort died.The number of deaths is rising quickly. As of Jan 24, 2020, 835 laboratory-confirmed 2019-nCoV infections were reported in China, with 25 fatal cases. Reports have been released of exported cases in many provinces in China, and in other countries; some health-care workers have also been infected in Wuhan. Taken together, evidence so far indicates human transmission for 2019-nCoV. We are concerned that 2019-nCoV could have acquired the ability for efficient human transmission.19Perlman S Netland J Coronaviruses post-SARS: update on replication and pathogenesis. Airborne precautions, such as a fit-tested N95 respirator, and other personal protective equipment are strongly recommended. To prevent further spread of the disease in health-care settings that are caring for patients infected with 2019-nCoV, onset of fever and respiratory symptoms should be closely monitored among health-care workers. Testing of respiratory specimens should be done immediately once a diagnosis is suspected. Serum antibodies should be tested among health-care workers before and after their exposure to 2019-nCoV for identification of asymptomatic infections.Similarities of clinical features between 2019-nCoV and previous betacoronavirus infections have been noted. In this cohort, most patients presented with fever, dry cough, dyspnoea, and bilateral ground-glass opacities on chest CT scans. These features of 2019-nCoV infection bear some resemblance to SARS-CoV and MERS-CoV infections.20Lee N Hui D Wu A et al.A major outbreak of severe acute respiratory syndrome in Hong Kong., 21Assiri A Al-Tawfiq JA Al-Rabeeah AA et al.Epidemiological, demographic, and clinical characteristics of 47 cases of Middle East respiratory syndrome coronavirus disease from Saudi Arabia: a descriptive study. However, few patients with 2019-nCoV infection had prominent upper respiratory tract signs and symptoms (eg, rhinorrhoea, sneezing, or sore throat), indicating that the target cells might be located in the lower airway. Furthermore, 2019-nCoV patients rarely developed intestinal signs and symptoms (eg, diarrhoea), whereas about 20–25% of patients with MERS-CoV or SARS-CoV infection had diarrhoea.21Assiri A Al-Tawfiq JA Al-Rabeeah AA et al.Epidemiological, demographic, and clinical characteristics of 47 cases of Middle East respiratory syndrome coronavirus disease from Saudi Arabia: a descriptive study. Faecal and urine samples should be tested to exclude a potential alternative route of transmission that is unknown at this stage.The pathophysiology of unusually high pathogenicity for SARS-CoV or MERS-CoV has not been completely understood. Early studies have shown that increased amounts of proinflammatory cytokines in serum (eg, IL1B, IL6, IL12, IFNγ, IP10, and MCP1) were associated with pulmonary inflammation and extensive lung damage in SARS patients.22Wong CK Lam CWK Wu AKL et al.Plasma inflammatory cytokines and chemokines in severe acute respiratory syndrome. MERS-CoV infection was also reported to induce increased concentrations of proinflammatory cytokines (IFNγ, TNFα, IL15, and IL17).23Mahallawi WH Khabour OF Zhang Q Makhdoum HM Suliman BA MERS-CoV infection in humans is associated with a pro-inflammatory Th1 and Th17 cytokine profile. We noted that patients infected with 2019-nCoV also had high amounts of IL1B, IFNγ, IP10, and MCP1, probably leading to activated T-helper-1 (Th1) cell responses. Moreover, patients requiring ICU admission had higher concentrations of GCSF, IP10, MCP1, MIP1A, and TNFα than did those not requiring ICU admission, suggesting that the cytokine storm was associated with disease severity. However, 2019-nCoV infection also initiated increased secretion of T-helper-2 (Th2) cytokines (eg, IL4 and IL10) that suppress inflammation, which differs from SARS-CoV infection.22Wong CK Lam CWK Wu AKL et al.Plasma inflammatory cytokines and chemokines in severe acute respiratory syndrome. Further studies are necessary to characterise the Th1 and Th2 responses in 2019-nCoV infection and to elucidate the pathogenesis. Autopsy or biopsy studies would be the key to understand the disease.In view of the high amount of cytokines induced by SARS-CoV,22Wong CK Lam CWK Wu AKL et al.Plasma inflammatory cytokines and chemokines in severe acute respiratory syndrome., 24He L Ding Y Zhang Q et al.Expression of elevated levels of pro-inflammatory cytokines in SARS-CoV-infected ACE2+ cells in SARS patients: relation to the acute lung injury and pathogenesis of SARS. MERS-CoV,25Faure E Poissy J Goffard A et al.Distinct immune response in two MERS-CoV-infected patients: can we go from bench to bedside?., 26Falzarano D de Wit E Rasmussen AL et al.Treatment with interferon-α2b and ribavirin improves outcome in MERS-CoV-infected rhesus macaques. and 2019-nCoV infections, corticosteroids were used frequently for treatment of patients with severe illness, for possible benefit by reducing inflammatory-induced lung injury. However, current evidence in patients with SARS and MERS suggests that receiving corticosteroids did not have an effect on mortality, but rather delayed viral clearance.27Stockman LJ Bellamy R Garner P SARS: systematic review of treatment effects., 28Lansbury L Rodrigo C Leonardi-Bee J Nguyen-Van-Tam J Lim WS Corticosteroids as adjunctive therapy in the treatment of influenza., 29Arabi YM Mandourah Y Al-Hameed F et al.Corticosteroid therapy for critically ill patients with Middle East respiratory syndrome. Therefore, corticosteroids should not be routinely given systemically, according to WHO interim guidance.30WHOClinical management of severe acute respiratory infection when novel coronavirus (nCoV) infection is suspected. Among our cohort of 41 laboratory-confirmed patients with 2019-nCoV infection, corticosteroids were given to very few non-ICU cases, and low-to-moderate dose of corticosteroids were given to less than half of severely ill patients with ARDS. Further evidence is urgently needed to assess whether systematic corticosteroid treatment is beneficial or harmful for patients infected with 2019-nCoV.No antiviral treatment for coronavirus infection has been proven to be effective. In a historical control study,31Chu CM Role of lopinavir/ritonavir in the treatment of SARS: initial virological and clinical findings. the combination of lopinavir and ritonavir among SARS-CoV patients was associated with substantial clinical benefit (fewer adverse clinical outcomes). Arabi and colleagues initiated a placebo-controlled trial of interferon beta-1b, lopinavir, and ritonavir among patients with MERS infection in Saudi Arabia.32Arabi YM Alothman A Balkhy HH et al.Treatment of Middle East respiratory syndrome with a combination of lopinavir-ritonavir and interferon-β1b (MIRACLE trial): study protocol for a randomized controlled trial. Preclinical evidence showed the potent efficacy of remdesivir (a broad-spectrum antiviral nucleotide prodrug) to treat MERS-CoV and SARS-CoV infections.33Sheahan TP Sims AC Graham RL et al.Broad-spectrum antiviral GS-5734 inhibits both epidemic and zoonotic coronaviruses., 34Sheahan TP Sims AC Leist SR et al.Comparative therapeutic efficacy of remdesivir and combination lopinavir, ritonavir, and interferon beta against MERS-CoV. As 2019-nCoV is an emerging virus, an effective treatment has not been developed for disease resulting from this virus. Since the combination of lopinavir and ritonavir was already available in the designated hospital, a randomised controlled trial has been initiated quickly to assess the efficacy and safety of combined use of lopinavir and ritonavir in patients hospitalised with 2019-nCoV infection.Our study has some limitations. First, for most of the 41 patients, the diagnosis was confirmed with lower respiratory tract specimens and no paired nasopharyngeal swabs were obtained to investigate the difference in the viral RNA detection rate between upper and lower respiratory tract specimens. Serological detection was not done to look for 2019-nCoV antibody rises in 18 patients with undetectable viral RNA. Second, with the limited number of cases, it is difficult to assess host risk factors for disease severity and mortality with multivariable-adjusted methods. This is a modest-sized case series of patients admitted to hospital; collection of standardised data for a larger cohort would help to further define the clinical presentation, natural history, and risk factors. Further studies in outpatient, primary care, or community settings are needed to get a full picture of the spectrum of clinical severity. At the same time, finding of statistical tests and p values should be interpreted with caution, and non-significant p values do not necessarily rule out difference between ICU and non-ICU patients. Third, since the causative pathogen has just been identified, kinetics of viral load and antibody titres were not available. Finally, the potential exposure bias in our study might account for why no paediatric or adolescent patients were reported in this cohort. More effort should be made to answer these questions in future studies.Both SARS-CoV and MERS-CoV were believed to originate in bats, and these infections were transmitted directly to humans from market civets and dromedary camels, respectively.35Cui J Li F Shi Z-L Origin and evolution of pathogenic coronaviruses. Extensive research on SARS-CoV and MERS-CoV has driven the discovery of many SARS-like and MERS-like coronaviruses in bats. In 2013, Ge and colleagues36Ge X-Y Li J-L Yang X-L et al.Isolation and characterization of a bat SARS-like coronavirus that uses the ACE2 receptor. reported the whole genome sequence of a SARS-like coronavirus in bats with that ability to use human ACE2 as a receptor, thus having replication potentials in human cells.37Wang M Hu Z Bats as animal reservoirs for the SARS coronavirus: hypothesis proved after 10 years of virus hunting. 2019-nCoV still needs to be studied deeply in case it becomes a global health threat. Reliable quick pathogen tests and feasible differential diagnosis based on clinical description are crucial for clinicians in their first contact with suspected patients. Because of the pandemic potential of 2019-nCoV, careful surveillance is essential to monitor its future host adaption, viral evolution, infectivity, transmissibility, and pathogenicity.ContributorsBC and JW had the idea for and designed the study and had full access to all data in the study and take responsibility for the integrity of the data and the accuracy of the data analysis. YWa, GF, XG, JiXu, HL, and BC contributed to writing of the report. BC contributed to critical revision of the report. YWa, GF, XG, JiXu, and HL contributed to the statistical analysis. All authors contributed to data acquisition, data analysis, or data interpretation, and reviewed and approved the final version.Declaration of interestsAll authors declare no competing interests.Data sharingThe data that support the findings of this study are available from the corresponding author on reasonable request. Participant data without names and identifiers will be made available after approval from the corresponding author and National Health Commission. After publication of study findings, the data will be available for others to request. The research team will provide an email address for communication once the data are approved to be shared with others. The proposal with detailed description of study objectives and statistical analysis plan will be needed for evaluation of the reasonability to request for our data. The corresponding author and National Health Commission will make a decision based on these materials. Additional materials may also be required during the process.AcknowledgmentsThis work is funded by the Special Project for Emergency of the Ministry of Science and Technology (2020YFC0841300) Chinese Academy of Medical Sciences (CAMS) Innovation Fund for Medical Sciences (CIFMS 2018-I2M-1-003), a National Science Grant for Distinguished Young Scholars (81425001/H0104), the National Key Research and Development Program of China (2018YFC1200102), The Beijing Science and Technology Project (Z19110700660000), CAMS Innovation Fund for Medical Sciences (2016-I2M-1-014), and National Mega-projects for Infectious Diseases in China (2017ZX10103004 and 2018ZX10305409). We acknowledge all health-care workers involved in the diagnosis and treatment of patients in Wuhan; we thank the Chinese National Health Commission for coordinating data collection for patients with 2019-nCoV infection; we thank the International Severe Acute Respiratory and Emerging Infections Consortium (ISARIC) for sharing data collection templates publicly on the website; and we thank Prof Chen Wang and Prof George F Gao for guidance in study design and interpretation of results.Supplementary MaterialReferences1.Richman DD Whitley RJ Hayden FG Clinical virology.4th edn. ASM Press,\n Washington20162.Ksiazek TG Erdman D Goldsmith CS et al.A novel coronavirus associated with severe acute respiratory syndrome.N Engl J Med. 2003; 348: 1953-19663.Kuiken T Fouchier RAM Schutten M et al.Newly discovered coronavirus as the primary cause of severe acute respiratory syndrome.Lancet. 2003; 362: 263-2704.Drosten C Günther S Preiser W et al.Identification of a novel coronavirus in patients with severe acute respiratory syndrome.N Engl J Med. 2003; 348: 1967-19765.de Groot RJ Baker SC Baric RS et al.Middle East respiratory syndrome coronavirus (MERS-CoV): announcement of the Coronavirus Study Group.J Virol. 2013; 87: 7790-77926.Zaki AM van Boheemen S Bestebroer TM Osterhaus ADME Fouchier RAM Isolation of a novel coronavirus from a man with pneumonia in Saudi Arabia.N Engl J Med. 2012; 367: 1814-18207.WHOSummary of probable SARS cases with onset of illness from 1 November 2002 to 31 July 2003.8.WHOMiddle East respiratory syndrome coronavirus (MERS-CoV).9.WHONovel coronavirus – China.10.WHONovel coronavirus – Thailand (ex-China).11.WHONovel coronavirus – Japan (ex-China).12.WHONovel coronavirus – Republic of Korea (ex-China).13.CDCFirst travel-related case of 2019 novel coronavirus detected in United States.14.Tan W Zhao X Ma X et al.A novel coronavirus genome identified in a cluster of pneumonia cases — Wuhan, China 2019−2020.15.Sanz F Gimeno C Lloret T et al.Relationship between the presence of hypoxemia and the inflammatory response measured by C-reactive protein in bacteremic pneumococcal pneumonia.Eur Respir J. 2011; 38: 249216.Kidney disease: improving global outcomes (KDIGO) acute kidney injury work groupKDIGO clinical practice guideline for acute kidney injury.17.Garner JS Jarvis WR Emori TG Horan TC Hughes JM CDC definitions for nosocomial infections, 1988.Am J Infect Control. 1988; 16: 128-14018.Gao C Wang Y Gu X et al.Association between cardiac injury and mortality in hospitalized patients infected with avian influenza A (H7N9) virus.Crit Care Med. 2020; ()19.Perlman S Netland J Coronaviruses post-SARS: update on replication and pathogenesis.Nat Rev Microbiol. 2009; 7: 439-45020.Lee N Hui D Wu A et al.A major outbreak of severe acute respiratory syndrome in Hong Kong.N Engl J Med. 2003; 348: 1986-199421.Assiri A Al-Tawfiq JA Al-Rabeeah AA et al.Epidemiological, demographic, and clinical characteristics of 47 cases of Middle East respiratory syndrome coronavirus disease from Saudi Arabia: a descriptive study.Lancet Infect Dis. 2013; 13: 752-76122.Wong CK Lam CWK Wu AKL et al.Plasma inflammatory cytokines and chemokines in severe acute respiratory syndrome.Clin Exp Immunol. 2004; 136: 95-10323.Mahallawi WH Khabour OF Zhang Q Makhdoum HM Suliman BA MERS-CoV infection in humans is associated with a pro-inflammatory Th1 and Th17 cytokine profile.Cytokine. 2018; 104: 8-1324.He L Ding Y Zhang Q et al.Expression of elevated levels of pro-inflammatory cytokines in SARS-CoV-infected ACE2+ cells in SARS patients: relation to the acute lung injury and pathogenesis of SARS.J Pathol. 2006; 210: 288-29725.Faure E Poissy J Goffard A et al.Distinct immune response in two MERS-CoV-infected patients: can we go from bench to bedside?.PLoS One. 2014; 9e8871626.Falzarano D de Wit E Rasmussen AL et al.Treatment with interferon-α2b and ribavirin improves outcome in MERS-CoV-infected rhesus macaques.Nat Med. 2013; 19: 1313-131727.Stockman LJ Bellamy R Garner P SARS: systematic review of treatment effects.PLoS Med. 2006; 3: e34328.Lansbury L Rodrigo C Leonardi-Bee J Nguyen-Van-Tam J Lim WS Corticosteroids as adjunctive therapy in the treatment of influenza.Cochrane Database Syst Rev. 2019; 2: CD01040629.Arabi YM Mandourah Y Al-Hameed F et al.Corticosteroid therapy for critically ill patients with Middle East respiratory syndrome.Am J Respir Crit Care Med. 2018; 197: 757-76730.WHOClinical management of severe acute respiratory infection when novel coronavirus (nCoV) infection is suspected.31.Chu CM Role of lopinavir/ritonavir in the treatment of SARS: initial virological and clinical findings.Thorax. 2004; 59: 252-25632.Arabi YM Alothman A Balkhy HH et al.Treatment of Middle East respiratory syndrome with a combination of lopinavir-ritonavir and interferon-β1b (MIRACLE trial): study protocol for a randomized controlled trial.Trials. 2018; 19: 8133.Sheahan TP Sims AC Graham RL et al.Broad-spectrum antiviral GS-5734 inhibits both epidemic and zoonotic coronaviruses.Sci Transl Med. 2017; 9eaal365334.Sheahan TP Sims AC Leist SR et al.Comparative therapeutic efficacy of remdesivir and combination lopinavir, ritonavir, and interferon beta against MERS-CoV.Nat Commun. 2020; 11: 22235.Cui J Li F Shi Z-L Origin and evolution of pathogenic coronaviruses.Nat Rev Microbiol. 2019; 17: 181-19236.Ge X-Y Li J-L Yang X-L et al.Isolation and characterization of a bat SARS-like coronavirus that uses the ACE2 receptor.Nature. 2013; 503: 535-53837.Wang M Hu Z Bats as animal reservoirs for the SARS coronavirus: hypothesis proved after 10 years of virus hunting.Virol Sin. 2013; 28: 315-317Article InfoPublication HistoryPublished: January 24, 2020IdentificationDOI: https://doi.org/10.1016/S0140-6736(20)30183-5Copyright© 2020 Elsevier Ltd. All rights reserved.ScienceDirectAccess this article on ScienceDirectClinical features of patients infected with 2019 novel coronavirus in Wuhan, ChinaFiguresFigure 1Date of illness onset and age distribution of patients with laboratory-confirmed 2019-nCoV infectionFigure 2Timeline of 2019-nCoV cases after onset of illnessFigure 3Chest CT imagesTablesTable 1Demographics and baseline characteristics of patients infected with 2019-nCoVTable 2Laboratory findings of patients infected with 2019-nCoV on admission to hospitalTable 3Treatments and outcomes of patients infected with 2019-nCoV\n \n\n \n\n\n \n\n \n\n \n\n Linked Articles"],["topImageUrl",null],["error",null]]}]}],["text","轉po:\n\n預估兩週潛伏期\nLancet論文統計指出\n\n兩成不發燒\n七成會咳嗽\n症狀非常不典型！\n\n一週後發病\n發病後一週\n32% 加護病房\n29% ARDS插管急救\n15% 死亡終結\n\n意思是\n重症一半會死！\n\n機率大概\n比六發子彈\n直接開一槍的\n俄羅斯輪盤低一些～\n\n中國官方還告訴大家沒事兒不要怕？！\n\n我身為\n高雄榮總\n急診一線醫師\n加護病房主治醫師\n我目前想不到有什麼其他比這個更嚴重更糟糕的傳染病了🙏\n\n\nLancet\n\nhttps://www.thelancet.com/journals/lancet/article/PIIS0140-6736(20)30183-5/fulltext"],["id","1fbhbmuiiyn7x"],["createdAt","2020-01-27T10:59:00.386Z"]]}]}],["relatedReplies",{"__iterable":"List","data":[{"__iterable":"Map","data":[["article",{"__iterable":"Map","data":[["replyCount",1],["references",{"__iterable":"List","data":[{"__iterable":"Map","data":[["type","LINE"]]}]}],["replyRequestCount",1],["user",{"__iterable":"Map","data":[["id","j4S8C_9GNQGlQwAl1lpQmVJcTheQRtm0SiyV2ggNVwYUoi4PA"],["name","斯文的三重桃樂斯"],["avatarUrl",null],["level",0]]}],["hyperlinks",{"__iterable":"List","data":[{"__iterable":"Map","data":[["title","Flipboard on Flipboard"],["url","http://flip.it/TDnC4H"],["summary","@flipboard | Official Flipboard accountYour hand gestures can help make you more charismaticResearch into public speakers suggests hand gestures can powerfully change the way you are perceived. David Robson explains.

This summer, as anger over the separation of migrant families at the border …Homeland SecuritySee How the Fall Equinox Is Celebrated Around the WorldCulture & ExplorationPhoto Gallery

From early-morning gatherings at Stonehenge to torch-bearing revelers parading through a city in Spain, people around the world herald the seasonal change in myriad ways.

This summer, as anger over the separation of migrant families at the border …Homeland SecuritySee How the Fall Equinox Is Celebrated Around the WorldCulture & ExplorationPhoto Gallery

From early-morning gatherings at Stonehenge to torch-bearing revelers parading through a city in Spain, people around the world herald the seasonal change in myriad ways.

As the autumn equinox approaches, days begin to shorten in the Northern Hemisphere, and people …National Parks"],["topImageUrl","https://cdn.flipboard.com/uploads/avatar/7e211185df7384dc5032a1b22b634616c56c1238.jpeg"],["error",null]]}]}],["text","廢墟、童話城堡都有！彰化8個人氣「免費」好拍景點\n\nhttp://flip.it/dqxAaY"],["id","5410467194193-rumor"],["createdAt","2016-12-27T04:55:00.000Z"]]}],["reply",{"__iterable":"Map","data":[["id","AV0bjelnyCdS-nWhubvf"],["user",{"__iterable":"Map","data":[["id","AVriAlOoyrDaTqlmmqhC"],["name","Lin"],["avatarUrl",null],["level",22]]}],["type","NOT_ARTICLE"],["text","旅遊景點分享。"],["reference",""],["hyperlinks",{"__iterable":"List","data":[]}],["createdAt","2017-07-07T05:37:51.466Z"]]}]]},{"__iterable":"Map","data":[["article",{"__iterable":"Map","data":[["replyCount",1],["references",{"__iterable":"List","data":[{"__iterable":"Map","data":[["type","LINE"]]}]}],["replyRequestCount",2],["user",null],["hyperlinks",{"__iterable":"List","data":[{"__iterable":"Map","data":[["title","Edward M. Mautner Oral History"],["url","http://www.jsc.nasa.gov/history/oral_histories/NASA_HQ/Experts/MautnerEM/MautnerEM_3-22-12.htm"],["summary","NASA Johnson Space Center \n Oral History Project\n Edited Oral History Transcript\n Edward \n M. Mautner\n Interviewed by Jennifer Ross-Nazzal\n Chantilly, Virginia – 22 March 2012\n Ross-Nazzal: \n Today is March 22, 2012. This interview with Ed Mautner is being conducted \n at the [Smithsonian National Air and Space Museum] Steven Udvar-Hazy \n Center in Chantilly, Virginia, for the JSC Oral History Project. Interviewer \n is Jennifer Ross-Nazzal, assisted by Rebecca Wright. Thanks again \n for taking time out of your schedule.\n Mautner: \n You’re welcome, it’s my pleasure to do it.\n Ross-Nazzal: \n If you would, tell us about the state of [OV (orbiter vehicle)-101] \n Enterprise in 2003 before she was restored.\n Mautner: \n My familiarity with Enterprise was quite limited before I began work \n on it. When we received it, it had been stored both outdoors and in \n a non-climate-controlled hangar since the mid-80s, and a lot of deterioration \n of the surface coatings—the paint, the markings—had taken \n place. Woodpeckers had gotten up onto the vertical stabilizer and \n pecked a lot of holes into the polyurethane foam that they used as \n a mockup for tiles. I don’t recall the number of holes, but \n there were quite a few of them. \n And then just about every form of creature had made the Enterprise \n their habitat and playground. Birds had gotten into it; bugs had gotten \n into it. Hornets or wasps had made their little mud nests in the landing \n gear wells. They made it out of this beautiful red Virginia clay dirt \n that surrounds the area. In fact, when we started to clean them it \n almost seemed as if the aircraft was bleeding, the soil they made \n those nests out of was so red.\n It had basically suffered a lot of inattention. It hadn’t been \n cleaned or worked on to any great degree. I believe it was outside \n for about two years prior to them building the hangar for it, and \n it had been rained on. There were many ports of entries for water, \n and so water had pooled inside the bottom of the fuselage and it had \n left a lot of staining from dirt, waterlines, etc. and also had caused \n some minor pitting and corrosion. Nothing that would weaken the structure \n of the aircraft but certainly had caused some deterioration.\n Ross-Nazzal: \n Tell us about that move, when you decided to move her from that hangar \n into this facility.\n Mautner: \n Until I walked in March 1st of 2004, I had never touched Enterprise \n and I had really nothing to do with it. I recall being in the hangar \n the day they moved it in, and just about all the employees who were \n out here took a break to watch them tug it down the tow way from the \n airport and into the building.\n What was noteworthy at the time was that they had cut a notch in the \n back wall of the space hangar in order to permit the vertical [stabilizer] \n to come through. It was the only way to get Enterprise into that building \n without taking the vertical off, which would have been a prohibitively \n difficult task. I came back the next day and the blanking plate had \n been restored as if the building had never changed at all, so that \n was pretty neat. It was a well-prepared move.\n That took place in the early fall of 2003, and while I was aware that \n it was in the building, it was sometime after January 2004 that I \n was notified that I would be working on it in preparation for the \n opening of the space hall [James S. McDonnell Space Hangar].\n Ross-Nazzal: \n What was your expertise prior to working on Enterprise? Were you working \n on aircraft restoration?\n Mautner: \n My background is—the British would describe it as checkered. \n I did a whole lot of things before I got directly involved in airplanes. \n I started out as a kid building model airplanes, and I never gave \n that up. I always built model airplanes, and I continue to do so. \n And I was always a museum rat, an airplane museum rat. Whenever we \n traveled, wherever we went, there was always a stop at an airplane \n museum somewhere.\n Somewhere around the late 80s, my wife and I had moved our children \n to southern California, and I began to volunteer in some aircraft \n museums out there. Planes of Fame in [Air Museum] Chino, and later \n at the Command Museum at U.S. Marine Corps Air Station El Toro, which \n was right in the town I lived in so it was convenient to work there. \n I started doing structures work.\n In January of 1989 I left a former career and made a great midlife \n career change whereby I wanted to work in an aircraft museum. I had \n enjoyed volunteering on Saturdays so much, I went to Cal [California] \n State University, San Bernardino, for a Museum Studies certificate \n program. My background was in auto mechanics, and I needed to create \n a resume to get into working on aircraft, especially in museums. I \n enjoyed it immensely but had a problem with the hours of class.\n By great good fortune, McDonnell Douglas [Corporation] was hiring \n mechanics in the summer of 1989. Actually they’d been hiring \n prior to that, but that’s when I found out. I went and I applied, \n and they hired me immediately because of my automotive experience. \n I became a structures mechanic on the C-17 [aircraft] program in Long \n Beach, California. That was my first real hands-on experience with \n real aircraft manufacturing.\n I worked at McDonnell Douglas from August of 1989 until June of 1994, \n when there were massive layoffs. They were losing their contracts \n for MD-80s [airliners], and the MD-11 [airliner] never took off. The \n C-17 was in a state of flux because Congress and the Air Force were \n at loggerheads, so I and tens of thousands of others were laid off \n during that year.\n Meanwhile I had gone from my certificate program at Cal State, San \n Bernardino, to a Master’s program at Cal State, Dominguez Hills \n in Historic Preservation. Because John [F.] McDonnell of the McDonnell \n Douglas Corporation was interested in an educated workforce, they \n paid for almost all of my Master’s program, everything but my \n research courses and my thesis. \n I had early on decided that my thesis was going to be in aircraft \n preservation at the National Air and Space Museum. I thought that \n they were the best. They were the ones that were writing about aircraft \n preservation, and I really wanted to write about how they did it, \n because that seemed to be the benchmark.\n In June of 1994 I was laid off, and I immediately contacted the museum \n to do an internship back here so that I could do further research \n for my thesis. In August and September of ’94 I did a 60-day \n volunteer internship. I was outside of their regular program, so there \n was no stipend; I just came in and volunteered. I worked in the restoration \n shop during the day until about 3:30 [p.m.]. I’d then go across \n the river, downtown to the main museum and I would work in the archives.\n The people in the archives—I don’t know why, I was a total \n stranger to them—they let me stay after hours. In fact, at five \n o’clock when they left, they issued me instructions on what \n lights to turn off and how to shut down the computers. I would stay \n there till 8:30 so I’d get all my research. I had one other \n paper and I had my thesis to do, so I spent almost every weekday night \n at the museum doing that.\n They said, “Stay in touch; we’re going to be hiring.” \n The hiring was going to ramp up for this facility, this new Udvar-Hazy \n Center, so I stayed in touch. Every month I contacted somebody associated \n with the restoration facility, and finally in 1997 they said, “Something’s \n going to happen. Get back here; be nearby.” So I stayed with \n relatives and I hung out and I volunteered, and finally in February \n of 1998 they hired me. They did it because I had the academic background, \n but I also had, from McDonnell Douglas and my volunteer efforts, the \n mechanical background as well. I was very fortunate that the timing \n worked out. \n My background is chiefly in aircraft structures, which would be the \n main metal pieces that form the basis of the aircraft upon which everything \n else is hung. After I get through with my work, then the hydraulics \n people and the fuel people and the electrical people and the power \n plant people take over.\n Ross-Nazzal: \n What sort of planes have you worked on since you began working here?\n Mautner: \n The first one that I started working on was a Japanese single-engine \n floatplane bomber, [the Aichi M6A1 Seiran]. It looks more like a fighter \n with floats, but it was in fact a strategic bomber that the Japanese \n designed to fold up into an 11-foot diameter circle and fit into the \n tube on a submarine. This submarine was an aircraft carrier, literally, \n and it carried three of these. Very significant aircraft, because \n it was designed to bomb the Panama Canal locks. \n The submarine was probably more technologically advanced than the \n aircraft. It was designed to go 27,000 miles without refueling, and \n they actually did have plans to use these small bombers to bomb New \n York [City, New York] and possibly even Boston [Massachusetts]. It \n was a metal airplane, and my sheet metal experience came to good use \n there. \n After that, they put me on a World War I bomber, a twin-engine fabric-and-wood \n bomber. That airplane was a Caudron G.4, a French bomber that the \n United States tested but never purchased. Because we had no combat \n aircraft in World War I, we had to go to the French and the British \n and the Italians. That was an excellent experience because that broadened \n out my sheet metal and structures experience to wood and fabric. That \n airplane is also currently on display here.\n The next aircraft is one I’m still working on. We have a limited \n number of people here, so we get shifted off of whatever project we’re \n on to take care of other things in the museum, such as creating a \n display or working on the Enterprise for nine months in 2004. From \n about 2000 until this year, I’m still working on a World War \n II German night fighter, the Heinkel 219, a huge metal twin-engine \n aircraft, where again my airframe and sheet metal experience has come \n into use.\n Ross-Nazzal: \n Tell us what lessons you learned from working on those aircraft that \n you applied to the Enterprise.\n Mautner: \n In my case, precious few. It was a great departure from what we call \n restoration, where you actually have to fabricate or replicate or \n repair structural pieces of an aircraft. This was largely a massive \n clean-up and painting effort, so it falls under the role of preservation. \n That’s really our role here, preservation. Everybody talks about \n restoration, but restoration is the court of last resort when everything \n has pretty much gone to pot and needs to be drastically repaired or \n replaced.\n What we did on Enterprise fell under the heading of preservation, \n but we called it a refurbishment because we didn’t do anything \n to repair it. It was still considered to be NASA’s aircraft, \n and it came to us in such good structural condition that there really \n was nothing to repair on it except for the paint work.\n Ross-Nazzal: \n Would you tell us about working with NASA and USA [United Space Alliance] \n on this project? I understand they were involved.\n Mautner: \n Yes, they were. When we first began on March 1, 2004, we were pretty \n much on our own. There were just three of us: myself, Anne [C.] McCombs, \n and Steve Kautner. We walked into this big empty hangar. The only \n thing that was in it at the time was Enterprise, and it didn’t \n even have a barrier around it because the public wasn’t allowed \n in that building yet. We just kind of stood there and wondered, “Where \n the heck are we going to start on this thing?” \n We had no high-lift equipment—all we had access to were ladders—so \n we decided to start working on the landing gear. We had no contact \n with NASA outside of certain parameters that they gave us in cleaning \n and painting it, that we were given by the curator, Valerie Neal. \n Without anything else but ourselves, we began the cleaning process \n at the lowest part of the Space Shuttle, areas that were accessible \n initially to us.\n Our experience with NASA and USA came a little bit later, and [their] \n big project was the removal of all of the cockpit windscreens, the \n front ones. I believe that there were three panels in them, and there \n were some engineering discussions as to whether they could get away \n with just two panels. The challenge for us was to provide them with \n the platforms to get up there and work off of. \n Anne and I were involved, and at that point we had a lot of mechanics \n from United Airlines [Inc.] come in to assist us. Their bosses permitted \n them to give us some high-lift platforms that were attached to trucks. \n We could maneuver a truck in and then raise a platform and extend \n it out over the nose of Enterprise in order for the USA guys to remove \n those windshields. The other problem was these windscreens were very, \n very heavy. They required quite a bit of effort to crane off of the \n Shuttle.\n At that point, I began to get to meet some of the guys, most specifically \n from USA. One of them was a fellow named Tom [Thomas W.] Roberts, \n and Tom’s my friend now. He was back here for the last visit. \n There were several other mechanics that came up from KSC [NASA Kennedy \n Space Center, Florida] who I would count as really good acquaintances, \n friends, who we worked with. Got to meet Kevin [C.] Templin. He was \n always there, very personable fellow, and always a go-to guy for information \n and leadership. \n Whenever they came back, we dropped what we were doing and became \n assistants to them, to the point where in this last visit we were \n almost considered more one of their group than as a separate entity, \n which was kind of neat. The things I saw from everybody in NASA and \n USA was enthusiasm, expertise, positive attitude, and great great \n teamwork. The first thing we noticed is how well they worked as a \n team. \n Many times we didn’t know that these guys had never worked together \n before, and yet it was a seamless effort. You got to talking to them, \n you got to know them, and you realized, this guy never worked on the \n back end of the aircraft. He always worked on the front and didn’t \n even know the guys that they were working with. That was inspiring \n to us to see that kind of effort that they used to put in.\n Some of my recollection of my efforts with NASA and USA get a little \n bit confused, because some of their efforts were during that nine-month \n period when we worked on preparing Enterprise for the opening of the \n space hangar, but several of them came later. They would ask Anne \n or myself or others from the previous team to come back and assist \n them. I believe that the windshield effort was made during that nine \n months we were there, but our other efforts with them came later, \n so they’d be outside the parameters of my work on the Enterprise.\n Ross-Nazzal: \n I also read that you had to replace the OMS [orbital maneuvering system] \n pods because they had rotted.\n Mautner: \n Yes, we did. One of the first things we did was to remove those. Tony \n [Anthony W.] Carp was the team lead for the lifting of the OMS pods. \n They were a large aluminum plate that mated up to the surface of the \n Shuttle, but everything above that aluminum plate was basically wood. \n Plywood flooring, then 2 x 4 and 4 x 4 framework, and then they put \n a fiberglass cover over the top of that.\n We discovered how weak the structure was when we started to tighten \n up the fittings for the lifting rings. We’d gotten a guy inside \n to tighten the nuts up inside because they seemed loose, and he kept \n tightening and tightening and tightening. We suddenly realized that \n wood was so soft from moisture that he was just tightening the nut \n and the washer right into the wood. We had to be very, very careful \n lifting those OMS pods off.\n The OMS pods were never planned to be on the Shuttle when the space \n hall opened. That was something that we were going to install later, \n so our job now was to focus just on the Shuttle, and guys back in \n the shop worked on the OMS pods.\n Ross-Nazzal: \n Are they on there now, have they been replaced?\n Mautner: \n They were put on I think in 2005, 2006 time range, and before the \n last visit to get Enterprise ready we removed them. They’re \n out in the shop, on the floor. The guys from USA put the ALTA [Approach \n and Landing Test Article] pods on. Those are the travel pods. They \n look like OMS pods, but they’re real metal. They’re real \n structural, flightworthy, and the tail cone fits up to those components.\n Ross-Nazzal: \n Tell us about cleaning and painting the vehicle. That sounded like \n a very large chore.\n Mautner: \n Yes, it really was. I started on March 1st to clean the right main \n landing gear bay and ultimately the landing gear and tires. Steve \n Kautner took the left bay and did the same, and Anne McCombs worked \n on the nose gear and the nose gear bay. It wasn’t until March \n 22nd that Tony [Carp] came on the team, and his job was to clean the \n entire vertical surface with the ultimate purpose of coring or filling \n the holes that the woodpeckers had made.\n Somebody might at this point ask, why would woodpeckers go after polyurethane \n foam? The explanation we got—I’m not an ornithologist—is \n that woodpeckers make the noise they do as a mating call, not just \n to get to bugs inside of the tree. Once they got through that polyurethane \n foam, they were banging on an aluminum surface. It made quite a noise, \n and it was far-reaching. There were about 75 holes ranging from very \n small, smaller than your pinky, to much bigger than your thumb. \n We were using, on NASA’s recommendation, Amway LOC [liquid organic \n cleaner] cut, one part LOC to ten parts water, to wash everything. \n It’s a mild organic detergent, and it’s biodegradable. \n This was what USA used, so this was what we used. And this is where, \n in the wheel wells, I found most of the hornet nests. Once you got \n the water into them they just started running down the beautiful epoxy \n white paint panels inside.\n At that point, once Steve had finished his wheel well, he started \n working on the bottom. He started up at the nose where he could reach, \n because you couldn’t get a ladder under there. He’s all \n scrunched up, just scrubbing away—and he was a bear of a guy. \n He had a lot of strength, and he just kept working down the bottom \n of the aircraft, all of the black-painted area of the aircraft. \n At that point we started using some lift equipment, JLG [Industries, \n Inc.] lifts and scissor lifts, to get up on the wing. We began cleaning \n the wing while Tony is cleaning the vertical, and we did this day \n after day after day. Then sometime in the summer, I believe in July, \n we began prepping the center fuselage for paint. The center fuselage \n does not have polyurethane foam on it, and that center fuselage extends \n from just behind the cockpit to back underneath the OMS pods. We also \n had to clean the payload bay doors. At this point we finally were \n able to get scaffolding in. We had a local company contracted to put \n scaffolding all up the sides and then bridge the top of the Shuttle. \n \n This is where things kind of got interesting. Initially it was doing \n just a lot of sanding on the side. Apparently the aircraft had been \n painted with aerospace quality polyurethane or epoxy paint and primer, \n but when it went on tour in 1984 it had started to look a little bit \n secondhand. They used a latex house paint to give it a quick repaint \n to make it appear better at the [New Orleans, Louisiana] World’s \n Fair here in the United States, and then they went to the Paris Air \n Show with it. This was all prior to delivering it to us.\n This paint over the years had become very brittle and very hard and \n would be quite a challenge to remove. Taking it off the sides was \n pretty easy. We were able to use electric sanders or pneumatic sanding \n devices, hand-held devices, and literally sand all of that down to \n the original paint. The sanding gave the surface a real good surface \n for paint to adhere to once we finished it and cleaned it up, but \n the real challenge came with the payload bay doors. We were instructed \n by NASA to not use any metal tools, any caustic chemicals, or any \n abrasives to remove the paint.\n We got up there, and they thought it would be really easy because \n a lot of the paint on the payload bay doors had flaked off, and some \n of it was literally sitting up. They said, “Just use duct tape \n and press it down real hard and pull it off, and that paint will pull \n up.” Well, that didn’t work at all. \n All of our communications with them were through Valerie Neal, the \n curator. The other thing we weren’t supposed to use was heat, \n so we went to Valerie and said, “The duct tape just isn’t \n cutting it.” They said, “Use plexiglas scrapers.” \n Plexiglas is a real hard plastic. We would cut up sections of it, \n rectangles of it, and then make a sharp edge at one end with a grinding \n wheel. That wasn’t cutting it either.\n One day I’m sitting on the scaffolding, trying to get this stuff \n off, and I see to my left this movement from about chest height over \n the curvature of the door. I look over, and it’s Steve Kautner \n with a metal scraper and a heat gun. I’m going, “Steve, \n what are you doing?” He goes, “This is the only way it’s \n coming off.” I said, “Okay, stop. Let’s talk about \n this.” \n We went back to our conservator, and I asked him, “Do you have \n a surface temperature gauge?” We went back and attached this \n temperature surface gauge to the area that we were putting the heat \n on, and we got some heat parameters. We were getting heat in the range \n of about 175 degrees [Fahrenheit] if we kept the heat gun moving, \n so we went back to NASA and they said, “Okay, you can do that, \n but no temperatures over 200,” or 250.\n All of a sudden this became easy, but we now found out very quickly \n why they didn’t want heat up there. The doors are made of carbon \n graphite, or carbon–carbon. It’s skinned with an almost \n aluminum-foil thickness coating of aluminum. It’s not like sheet \n metal, it’s more like a foil, and if you left the heat on there \n too much it would blister. We got a couple of thumb-sized blisters; \n we damaged the door. This concerned us, and we kept talking to each \n other. We’d just make sure we’d keep that heat gun moving \n and don’t overheat one spot.\n It became very evident very quickly why they didn’t want heat \n up there. They had done the same thing at some time in the past, only \n their blisters were much bigger than ours. That told us a) it’s \n been done this way before, and b) there’s a repair for it. We \n did make a few more blisters, an inevitable result of what we were \n doing, but we knew that if they needed those doors, they could be \n made flightworthy. \n Then, because there was still a lot of paint residue clinging to the \n surface, we got down and used acetone. We were all garbed up with \n double-canister air masks. There’s terrific pictures of us all \n down on our hands and knees on the door, all tied off because we’re \n high up on the top of the Shuttle, scrubbing with Scotch-Brite scrubbers \n and acetone to get the surface completely clean and ready for paint.\n That was another thing they told us, “You can’t stand \n on the doors.” So we had these four bridges across, and we’re \n all leaning off these bridges and working off of them. Again, Steve \n Kautner’s the first one to step off right on the doors. There’s \n no other way to get out there, there was no way. I said, “Okay, \n stop. Let’s talk this over.” We went down inside the Shuttle, \n and we looked up at the doors and we could see the latching system. \n \n We knew that those doors in zero gravity could open without any support, \n but we knew that they had big supports on them when they opened them \n in the hangar down in KSC. We looked at those supports, and we looked \n at how the doors fit together. It’s built like a bridge as long \n as they’re closed and locked, so four, five, six of us all were \n out on our hands and knees on the doors, scrubbing away to get the \n last residue and make a really good surface for paint.\n That brings up another aspect for me. I couldn’t wait every \n single day to come in. I’m not a space guy, I’m an airplane \n guy. In truth, the Shuttle is probably more like an aircraft than \n a spacecraft, and that one [Enterprise] never did go into space so \n it really was just an aircraft. I never followed space, I never got \n excited about it, but I got excited by this project. I couldn’t \n wait to get in every single day. Every day was interesting and exciting, \n and we were making progress—slow—but we were making progress. \n \n When they put the scaffolding up in July, we now had to climb this \n scaffolding. We had our harnesses on and we had our tethers, but we \n couldn’t tether off anywhere on the way up. We’re climbing \n 30, 35, almost 40 feet in the air every morning, every break time, \n every time you had to leave, and then when we got up there we would \n tie off and secure ourselves. But that little element of danger made \n it exciting, and we talked about that a lot too.\n We had two injuries, but only one on Enterprise. One of our guys did \n abrade his hand, but working on something else while we were out there, \n nothing to do with the Shuttle. One injury, very minor, on the Shuttle. \n One of our members fell off the payload bay doors, but he was tethered \n and it stopped him, and all he did was bang up his knee. I think we \n lost him one day, but that was it. The fact that we talked almost \n every day about the safety issue, that he was tethered off—we \n didn’t have more issues. \n The other thing that made it interesting and fun is that we were on \n this huge, very obvious artifact in the middle of this totally empty \n hall, and the public could come in to the front of the hall—there’s \n an overlook up above—and watch us. I think the element of being \n on stage made it exciting and interesting, too.\n There were some stresses. We always seemed short of personnel. At \n one point I realized there were a whole bunch of guys working on the \n OMS pods, and I had to convince my management that the OMS pods aren’t \n needed by October 19th, but the Shuttle is, so can I have more guys? \n And I got more guys.\n Once we had sanded down the sides and had removed the paint from the \n doors, it was time for a final prep and primer and paint. We had four \n guys do that, and we had a couple of issues that we had to deal with. \n One, it had to be an aerospace quality paint. PPG, Pittsburgh Paint \n and Glass, has always supplied us with our paint system that we’ve \n used exclusively for 20 years back at the [Paul E.] Garber [Preservation, \n Restoration, and Storage Facility, Suitland, Maryland], our restoration \n shop. Dave Wilson got in touch with them, and they turned him over \n to their aerospace division. They donated what amounted to about $15,000 \n worth of white polyurethane aerospace paint and primer.\n Our next challenge was we could not spray in the building because \n the building had an open port, and some of the ventilation systems \n functioned with the aircraft hangar. Also, the filters in the ventilation \n system would get clogged up with any paint residue and dust, so we \n had to either brush or roll it. Dave talked with the people from PPG, \n and they had an additive that allowed the paint to flow so that it \n would not appear to be rolled-out paint. It wouldn’t be textured, \n it would be nice and smooth.\n We got the paint on September 17th—keeping in mind that this \n hall is supposed to open on October 19th of 2004. They had to rest \n assured that all the fumes would have dissipated by 10 o’clock \n opening the next day, so we painted at night. It was Tony, Dave, myself, \n and a fellow by the name of Bob Weihrauch who did it. We would come \n in, and that hall was completely black except for the Klieg lights \n that we worked under. We rolled out the primer, and then we rolled \n out the white. [We rolled out two coats of primer and two coats of \n white.]\n I love working here at night because there’s no people here, \n the lighting is all dim, and it’s like going into a great cathedral \n at night when nobody’s around. It’s just beautiful. It’s \n quiet. You’ve got all these objects that you love, many of which \n you’ve worked on, and there’s just a neat thing about \n working here at night. We had a really good time. We had a really \n good crew. I don’t recollect ever having any arguments or disagreements \n or problems, we just really had a good time.\n One outstanding recollection is this facility is used by organizations—police \n organizations, FBI [Federal Bureau of Investigation], the MWAA, Metropolitan \n Washington Airport Authority. They have their own SWAT [special weapons \n and tactics] team and security forces. They do a lot of work in here \n at night, but we didn’t know that. One night three of us are \n up on this scaffolding, and Dave Wilson is on a scissor lift bringing \n paint up to us, and all the sudden there’s movement in the hangar. \n \n Keep in mind that outside of this cone of light that we’re in, \n everything’s dark out there, but there’s movement. We \n look down, and there’s 20 or 30 guys in black helmets, black \n outfits carrying guns, and they’re swarming like ants through \n our hangar. Well, I immediately was not concerned because none of \n them are looking at us or pointing the gun at us. We’re in white \n bunny suits, paper coveralls, and the Klieg lights are on us. We’re \n out there, very obvious.\n One guy kind of moved up to Dave, and he goes, “Have you seen \n her?” And Dave goes, “Seen who?” He goes, “Have \n you seen her?” and then he had to move, because you’re \n not supposed to look for hints. Apparently they were on an exercise, \n and one of their supervisors came in two hours ahead of time and hid \n herself somewhere in the museum, and they had to find her. Nobody \n told us; our security had not advised us that this was going to happen. \n \n Another time, during the day, I was working on the Shuttle and nobody \n else was around. Security came up to me and said, “We have a \n congressman or a senator here with his daughter back from college, \n and he’s got his office manager and his security detail. Would \n you come down and talk to them?” So I walk inside of this circle \n of Secret Service people, and they’ve got their radios and they \n all look very, very official, and I started to talk to them. I said, \n “What state do you represent, sir?” He goes, “Oh, \n one of your neighbors.” Meanwhile, his handler, his office guy, \n did all the talking. \n I went back that night and I looked all over the Internet for our \n neighboring senators and congressmen, and I did not find him—because \n you can get the photos of all of them. I went back to security the \n next day and I said, “What was that all about?” He says, \n “Oh, it’s just a test.” It was Secret Service in \n a practice run, and these guys were just members of the Secret Service \n that were playing a role to make it seem realistic. There are a lot \n of activities that go on here that are very interesting. But again, \n I was not forewarned of that. \n The other two events that were really interesting—Joe [H.] Engle, \n one of the test pilots for Enterprise, came back and spent a day with \n us. He is just a prince of a guy, just a super guy. We got to tour \n him around and show him what we were doing, and he was just the friendliest \n guy and was very very grateful to us restoring “his” bird \n back to its original condition. A couple weeks later, we all got an \n 8 x 10 of him standing by Enterprise out at Palmdale [California] \n back in’77, ’78, with a beautiful handwritten note and \n one of the original patches from his missions. It was just neat, a \n neat thing. \n The other very very interesting visitor to us was Colonel Pamela Ann \n Melroy. She’s a two mission pilot and one mission command pilot \n [commander]. She was also on the Columbia [STS-107 accident] investigation \n team, and she led the cockpit reconstruction area. My recollection, \n she was about five-foot-five, five-foot-six. I immediately made a \n connection, because she was a test pilot on the C-17 program. As I \n had worked on it, we had some things to talk about. \n I got to tour her around, and the thing I recollect so well is that \n she made a comment when we were in the cockpit. The cockpit of Enterprise \n is for the most part empty. The dashboard is there, but there’s \n only about three instruments, and they’re all old analog instruments. \n The seats are not there. She looked at that dashboard and marveled \n at the fact that it was the first time she’d seen one in a long \n time that wasn’t bits and pieces, because she had been looking \n and reviewing and trying to reassemble pieces of Columbia’s \n instrument panel.\n This diminutive colonel in the Air Force—always at the end the \n photographer comes in and takes a picture of us with the VIP [very \n important person]. Anne and I were there at that time, and we’re \n all standing off to the side, a foot away or something. She says, \n “Hey, we’ve been together all day. Come here!” She \n pulled us in right next to her to take the picture. We also got an \n 8x10 from her, signed, with a thank you and mission patches. Those \n are two of my favorite treasures from that whole time there. \n Ross-Nazzal: \n Nice recollections of that time period. \n Mautner: \n Yes, it really was. We got it done. Everything seemed late—getting \n personnel, the scaffolding came late, the paint came late—but \n we got it done. We got it done on October 18th, and October 19th was \n the day the media and the museum VIPs and everybody showed up to see \n it. It was a moment of pride because it really looked good. \n Tony and I stayed after October 19th. Tony had some more work up on \n the vertical to do, and the one area that we hadn’t really addressed \n was the propulsion bay. I would work in the propulsion bay; he’d \n work up top. We did this at night because the gallery was open to \n the public, so we did a lot of night work from October until we went \n away on Christmas vacation.\n Most of cleanup in the propulsion bay—there was some FOD [Foreign \n Object Debris], there was old hardware. It went back to when it was \n delivered to us. There were some sections of insulating aluminum, \n nuts, bolts, washers, things like that. A lot of water had been back \n there, and the floor panel structure was a huge, thick piece of aluminum \n that they had milled out to look almost like a waffle. The floor looked \n like a large waffle pattern, so there’s all these thousands \n of little pockets that had held water, and I had to clean them.\n We had some nights where we had events, and the events usually allowed \n people to roam into the space hangar, so I would take my knapsack \n and my coffee and my lunch, my snacks, up into the propulsion bay. \n Something you don’t normally do around artifacts, but I didn’t \n think I could come out because of the events they had going on. I’d \n stay up there for eight hours. Well, there’d be a potty break.\n We finished before our Christmas break, and then both of us reported \n back to the Garber Facility the first week in January 2005. It was \n rewarding every day, but they also rewarded our team with the Museum’s \n Peer Award. The [museum] gives two Peer Awards every year, and the \n Enterprise Preservation Team was awarded [one of them] for 2004. It \n was a great experience.\n Ross-Nazzal: \n What are you working on now that Enterprise is getting ready to take \n a new journey in its history to go to the Intrepid [Sea, Air & \n Space Museum in New York City]? Are you working on that?\n Mautner: \n I did work on it. I don’t know what my role will be in April \n when they come back, but the whole month of February I was assigned \n out here, along with Anne and one or two other folks, to escort NASA, \n USA, and some folks from [the] Boeing [Company]. “Escort” \n has a kind of static sound to it. We were essentially here to open \n doors that they couldn’t get into, including getting to the \n restrooms, and we were able to get them discounts in our shop and \n at McDonalds. \n And then pretty soon it was, “Oh my gosh. We don’t have \n this; do you have this tool?” So we’re running for tools. \n Later, as our professional relationship evolved, we were forklifting \n equipment for them. They had forklifts, but we were able to help them \n with that. We had a JLG man lift that we were able to take them and \n some of their equipment up to different places on the Shuttle, and \n also on the tail cone, which they assembled right out here in this \n shop. As to my role when they come back on the 19th, I have not been \n told what that is yet. I hope [I] do, I have some friends I’d \n like to hang out with again.\n Ross-Nazzal: \n Are you preparing in any way to accept [OV-103] Discovery, or are \n you primarily working with the Enterprise?\n Mautner: \n I’m not. My responsibility is to work on this German night fighter. \n We still have a wing to complete. The airframe fuselage is out in \n the museum right here now, but the wing has to be done and a lot of \n other details. Between escort duties and whatnot, I’m working \n on that. We’re told that with everything else that’s going \n on, we’re going to be moving our shop from Maryland out here. \n I don’t know how that’s going to work out with the Discovery–Enterprise \n exchange, that’s yet to be determined.\n Ross-Nazzal: \n It must have been rewarding to receive that from your peers, that’s \n wonderful.\n Mautner: \n Very, yes. I think Valerie Neal, former NASA and our curator, was \n probably instrumental in getting that for us. It was pretty neat.\n Ross-Nazzal: \n Rebecca, did you have any questions for Ed?\n Wright: \n Just one, and I think you might have already answered it. Did you \n ever come back and do anything else with Enterprise?\n Mautner: \n I have once or twice, but it was generally just escort duty. Those \n were times where it was just sit around and wait for whatever the \n folks, predominately USA, needed. I did come back, and we did reinstall \n the OMS pods. That was pretty much the original team again that did \n that, and we brought a couple of other people in to assist because \n you need spotters and tag line holders. \n Wright: \n You mentioned that you would escort for whatever they needed. What \n were the types of things that they would be coming back for?\n Mautner: \n Every time USA came here, they had a lot of supplies. Rather than \n transport them back, they would leave them out and we would store \n them in our barn, which is out in the back corner of the property. \n So the first thing we’d have to do is get their supplies, and \n that ran anywhere from tape to paints to epoxy glues.\n Recently, within the past year, they came back and we took them up \n on a JLG man lift. They did the work, and I would just maneuver it \n around to a position that helped them. Like I said, “escort” \n sounds very static, and sometimes it is, but a lot of times there’s \n a dynamic component, which is the most fun of all. That’s how \n you get to know these guys and get to see what they do.\n Wright: \n Were they doing types of restoration themselves?\n Mautner: \n Yes, about last April they came back to begin the process of making \n Enterprise airworthy. That involved making an assessment of all systems, \n and they also did the [landing gear] retracts. The main job was to \n get the hydraulic system working so they could retract the landing \n gear. Part of our position as escort was to shut down areas of that \n hall to keep the public away, because the machine, what they call \n the mule, fires up the hydraulics system, makes a lot of noise. Also \n there’s part of the retract that there’s noise involved \n as well.\n We did a lot of work stanchioning off. Moving stanchions back, and \n then when it was all done, moving the stanchions closer. We were trying \n to do this juggle between making sure people were safe and USA had \n enough room to work in, and also still keeping the hall open to the \n public. Our whole purpose is to let them get as close to this stuff \n as we can.\n There were a couple of times where, yes, it was pretty static, but \n other times we were pretty involved in helping them. And everything \n back here is secured. You can’t get into the restrooms without \n our badge, so we were escorting to the restrooms. Eventually, and \n especially in this last trip in February where they did the final \n preparation for flight, the security office provided them with badges \n that allowed them to open all the things they had to get into. They \n had the same access employees had. \n I think that speaks of the type of relationship that we’ve always \n felt we had with NASA, that they weren’t an alien organization \n that comes in and invades our space but rather cohorts, colleagues \n that we feel—not exactly one with, our roles are different—but \n certainly very good friends with.\n Wright: \n Is this a very unique exhibit here compared to all the rest? Do you \n have any other kind of associations with other groups that come in \n and work on their planes?\n Mautner: \n No, there’s no other organization that has the relationship \n that NASA has with us. I think that’s historically because we’ve \n been the repository of first choice for NASA, but also because Valerie \n Neal, who came from NASA, has maintained a very, very good relationship \n with all the leadership there. It’s been a very collegial relationship \n from the very beginning. Because of that, your relationship personally \n to these people is much more relaxed, so it’s much much easier \n to make friends. I have a couple buddies that I email once in a while \n back at the Cape [Canaveral, Florida]. It’s a good relationship.\n Ross-Nazzal: \n Any other last thoughts? \n Mautner: \n I think I’ve covered the important aspects of my relationship \n with the Shuttle Enterprise and with the folks from NASA and USA. \n We had a good team. I think it was kind of neat because management \n left us alone; they left us to make a lot of our own decisions. Everybody \n worked with the goal in mind, and we reached the goal. We finished \n Enterprise, it really looks beautiful. It took us nine months to get \n it in the shape that it’s in right now, and I have to say I \n am quite proud of how it came out. \n I do recollect one of the stories that I’ll tell you. When we \n began to strip all the paint off, we traced all of the markings—the \n NASA worm, the “United States” on the side, the “Enterprise” \n itself—so that after we painted it we could restore those. We \n noted and photographed the fact that these had been hand-brushed. \n Somebody had done a stencil, and then they hand-brushed in the black \n color. As we began to strip the aircraft, we realized that there were \n other markings, other NASA worms and another “Enterprise” \n and another “United States” underneath that exterior coat \n of latex house paint. \n We traced those as well, and at that point the curator had to make \n a decision. Are we going to restore it to when it was originally painted \n and lettered, or are we going to paint it as it was presented to us \n in 1985? The decision was made to paint it as it was delivered to \n us. Those letters were all in black, and the letters we found underneath \n were in lighter shades of dark gray. \n At the time that we traced them back on the aircraft, the question \n was asked, “Well, who feels comfortable painting them back on?” \n I mean, none of us are professional sign painters. Out of the five \n of us there, four chose to do it. So what you’ll see out there \n is our hand-lettered letters, and they are letter perfect. They came \n out really, really well. Three employees and one volunteer painted \n them all in. That too raised our level of pride and involvement and \n attachment to the Enterprise. \n I have mixed feelings about it going. On the one hand, it’s \n very important for us to have an orbiter, and Discovery is a much \n more significant vehicle to have in our collection, so I’m happy \n about that. On the other hand, all of our handwork is being sent off \n to somebody else. I’m not going to lose sleep over it, but I \n will think, how are they taking care of it? Is that beautiful paint \n job still squeaky clean and nice, and are all those tiles in good \n condition? So yes, I am concerned to some degree at it going off to \n other hands.\n Ross-Nazzal: \n Any plans to go to New York and see her once she’s in her permanent \n location?\n Mautner: \n No plans at this time, no.\n Wright: \n And will you be working on Discovery when it comes?\n Mautner: \n We’re not going to touch Discovery. This is another aspect that’s \n pretty neat about getting Discovery. The ideal airplane or spacecraft \n to get into our collection is the one that just went on its last mission. \n If we could get a Boeing B-17 [aircraft] that flew a mission from \n England in August or September of 1944, and then was transferred right \n back into our hall out front, that’s the idea. You don’t \n want to touch it. You want to mount it in such fashion that you preserve \n the tires and take any other stresses out of it, but it would be great \n if it had the gunpowder stains and the engine stains and the oil stains \n and the rough-hewn patches that the mechanics put on the planes back \n in those days. \n And that’s how Discovery is coming to us. They haven’t \n cleaned it, outside of taking out the toxic materials in all the tanks \n and systems, so it’s not going to look like Enterprise at all. \n In fact, one of the people from NASA commented that Enterprise looked \n like a toy. I think what she meant was it looked like a model, and \n Discovery looks like a real orbiter. So we’re happy about that.\n Wright: \n Well, thank you.\n Ross-Nazzal: \n Thank you for spending some time with us this morning. We appreciate \n it.\n Mautner: \n You’re very welcome. It’s my pleasure to do it.\n [End \n of interview]\n \n \n Return \n to JSC Oral History Website"],["topImageUrl","https://www.jsc.nasa.gov/history/oral_histories/head_images/sc.gif"],["error",null]]}]}],["text","美國太空總署員工表示，\n他們是使用太空總署建議的\n【安麗 LOC】以「1:10」濃度\n拿來清洗太空梭及所有東西。\n\n這就是表示該產品是【極度安全】的！\n\n這樣，還有什麼安麗產品是需要擔心呢？\n\n\nhttp://www.jsc.nasa.gov/history/oral_histories/NASA_HQ/Experts/MautnerEM/MautnerEM_3-22-12.htm"],["id","5484461227031-rumor"],["createdAt","2017-01-11T01:17:00.000Z"]]}],["reply",{"__iterable":"Map","data":[["id","AV_TT0CjyCdS-nWhujcr"],["user",{"__iterable":"Map","data":[["id","AV_TJNx9yCdS-nWhujbL"],["name","Victor Cheng"],["avatarUrl","https://www.gravatar.com/avatar/b0924f092282f5a2cc895e7f4b4c9cbc?s=100&d=identicon&r=g"],["level",4]]}],["type","NOT_ARTICLE"],["text","送出文章的人在嘗試與機器人聊天。\n未指定所謂“安全”之範疇，可以拿來洗東西不代表可以拿來吃。\n同理一間公司的其中一項產品安全也不代表所有東西安全，就像Samsung s7好棒棒，不代表Note 7不會爆炸。"],["reference",""],["hyperlinks",{"__iterable":"List","data":[]}],["createdAt","2017-11-19T08:05:07.366Z"]]}]]},{"__iterable":"Map","data":[["article",{"__iterable":"Map","data":[["replyCount",1],["references",{"__iterable":"List","data":[{"__iterable":"Map","data":[["type","LINE"]]}]}],["replyRequestCount",1],["user",{"__iterable":"Map","data":[["id","j4S8C_edDdU96oZupq3Xms9tKo2siS2uA7HMWfNP6BYlD1P_g"],["name","Oo好學上進的下營克莉絲汀oO"],["avatarUrl",null],["level",0]]}],["hyperlinks",{"__iterable":"List","data":[{"__iterable":"Map","data":[["title","キャンペーンに賛同！"],["url","https://www.change.org/p/request-permanent-court-of-arbitration-to-investigate-unlawful-use-of-force-of-hong-kong-police?utm_content=cl_sharecopy_17005953_en-GB%3Av5&recruiter=90670311&recruited_by_id=0040d850-5bec-43c4-8e81-c03a3953c79d&utm_source=share_petition&utm_medium=copylink&utm_campaign=psf_combo_share_message&utm_term=psf_combo_share_initial&share_bandit_exp=message-17005953-en-GB&share_bandit_var=v0"],["summary","(Update on 11/11/2019)\nURGENT! \nHong Kong need help from the World ! Please sign and support ! \nWe are writing to request Permanent Court of Arbitration (PCA) to formally Investigate Hong Kong Police for Unlawful Use of Force against Citizens since 9 June 2019\nAttn: Mr. Hugo H. Siblesz (Secretary-General of Permanent Court of Arbitration)\nDear Mr. Hugo Siblesz,\nWe are Hong Kong citizens. Hong Kong urgently need helps now!\nWe are writing to ask for your help to formally investigate police brutality against citizens of Hong Kong in the recent protests since 9 June 2019. A group of international experts appointed to help Hong Kong’s police watchdog conduct a study into the ongoing protests has said the body lacks power and urged for a more comprehensive investigation by an independent group with requisite powers.https://www.facebook.com/800660560021071/posts/2581953451891764/\nThe Hong Kong police front line police is getting insane. After the death of a 22-year-old student who died near a police clearance operation of a protest on last Friday. On 10 November, multiple arrests being made in Festival Walks, a shopping mall, and chaos scattered everywhere on the streets and in different housing estates.\n11 November 2019 is absolutely chaos. It is reported that the real guns shot by the police officers in different public areas.\nA Hong Kong police officer has shot a black-clad protester in the stomach at close range :\nhttps://www.facebook.com/800660560021071/posts/2583690221718087/https://www.facebook.com/800660560021071/posts/2583667861720323/\nA Hong Kong police motorcyclist appears to \"ram\" a crowd of black-clad protesters on 11 November:https://www.facebook.com/800660560021071/posts/2583726308381145/\nHundreds of thousands of people in Hong Kong took to the street in recent days to protest amid the controversial Extradition Bill. In the latest rounds of the protest, the Hong Kong Police Force (HKPF) has been using excessive level of violence and firearms against citizens who participated in peaceful protests. Confirmed crowd control weapons and ammunition in use including batons, pepper spray, rubber bullets, sponge grenades, live bullets, water cannon, tear gas. The sole purpose of this level of extreme violence “intent to injure”. It is believed to be a direct and flagrant violation the “UN Principles on the Use of Force and Firearms by Law Enforcement Officials.” Many unarmed protesters have been reportedly beaten or shot, some directly in their heads.\nSince 9 June, we believe that Hong Kong Police Force had committed the following crimes:\n1) Crime of aggression2) Conventional War Crimes3) Crimes against Humanities\nHong Kong’s anti-government protesters have sustained for five months. Here are some figures:\n1) It is 156 days of unrest since June 9\n2) 3,300+ protest-related are currently arrested, 10+ people are dead\n3) 6,200+ tear gas canisters used\n4) 15 bullet+, 2,000+ rubber bullets, 670+ foam bullets and 450+ beanbags bullets were fired\nRecord of the Hong Kong Police Brutality are as Follow:\n(A) Hong Kong Police Force Investigation Reporthttp://hkpfreport.org/\n(B) Hong Kong Police Violence Database:http://tl.hkrev.info/en/police-timeline/\n(C) Unlawful use of force by Hong Kong Police https://www.amnesty.org/en/documents/asa17/0576/2019/en/\n(D) Allegations of Hong Kong Police Force misconduct surrounding the 2019 Hong Kong protestshttps://en.m.wikipedia.org/wiki/Allegations_of_Hong_Kong_Police_Force_misconduct_surrounding_the_2019_Hong_Kong_protests\n(E) A Report of the 2019 Hong Kong Protest (written by Martin Purbrick, a former Royal Hong Kong Police officer whose service included Special Branch engaged in counter-terrorism intelligence)https://bit.ly/2nVuglA\n(F) The Six-monthly Report on Hong Kong - 1 January to 30 June 2019 (by Parliament Secretary of State for Foreign and Commonwealth Affairs)https://assets.publishing.service.gov.uk/government/uploads/system/uploads/attachment_data/file/843492/Hong_Kong_Six-monthly_Report_Jan-Jun19.pdf\n(G) Weapons of mass control, tactics of mass resistancehttps://www.reuters.com/investigates/special-report/hong-kong-protests-violence/\nFourteen Unlawful Use of Force by Hong Kong Police Force\n1) Shoot and attack both protesters and reportersUse live bullets dangerously. In October, a teenager was shot by wielding a pipe with live ammo at a point blank range with a revolver.https://youtu.be/PN4MvOrPotkAn Indonesia reporter was shot by police in rubber bullet on the face and become permanently blind in one eye.https://youtu.be/gWn5oRJKiM8https://www.bbc.com/news/world-asia-china-49910636\n2) Unlawful use of batonsPolice officers use batons to beat people, no matter they are protesters or un-involved bystanders. Police would keep beating people with batons while they lay on the ground with no resistance. That was inconsistent with the use of force guidelines that was taught in Police College.https://youtu.be/kW4A9hy3H3Y\n3) Improper use of riot control agents(RCA) - tear gas, pepper spray and water cannon. Use pepper spray on a person who posed no obvious threat is usual.https://youtu.be/4QvQTW40exYhttps://youtu.be/TAzkH4EncPA\n4) Arbitrary arrests people The arresting is from age 10 to 82. The police after failing to arrest the radical protesters, turned to arrest peaceful protesters, bystanders, young people in general and people wearing black clothes in young age, and even doctor.https://youtu.be/LnUFOjHYuxYhttps://youtu.be/yLbaXMaaOq4https://youtu.be/C_h1oJYyTe4\n5) Lack of visible identificationDuring many of the Hong Kong Police operations, the HKPF did not wear any ID badges nor did they produce any warrants. They had infringed the SOP (Standard Operating Procedure, and neglected the ROE (Rule of Engagement).https://www.hongkongfp.com/2019/08/23/tactical-squad-may-hiding-ids-order-protect-privacy-says-hong-kong-police-watchdog-vice-chair/\nhttps://www.scmp.com/news/hong-kong/law-and-crime/article/3017775/hong-kong-police-accused-provoking-protesters-and\n6) Restrictions on journalists and medicsJournalists:https://www.scmp.com/news/hong-kong/politics/article/3026940/press-associations-call-hong-kong-police-curb-attacks\nhttps://www.scmp.com/news/hong-kong/politics/article/3026940/press-associations-call-hong-kong-police-curb-attacks\nhttps://www.hongkongfp.com/2019/10/04/aggressive-obstructive-hong-kong-journalists-association-files-legal-challenge-police-treatment-press-protests/\nMedics:https://youtu.be/x_P0lBpgscs\nhttps://www.scmp.com/news/hong-kong/politics/article/3024947/hong-kong-police-apply-search-warrant-access-medical\n7) Brutal beatings, torture and sexual abuse protesters in police detention. Sexual Abuse in the police’s San Uk Ling Detention Centre (dark terror cell) which is located in an area close to the China-Hong Kong Border. It was reported that some students were tortured after arrest, leaving bone fractures. The police officers in the terror cell was “unnamed”, “unverified” and “unnumbered” and the cell has no cctv systems for recording what happened. The terror cell after the public decry is no longer used for detention. https://www.amnesty.org/en/latest/news/2019/09/hong-kong-arbitrary-arrests-brutal-beatings-and-torture-in-police-detention-revealed/?utm_source=TWITTER-IS&fbclid=IwAR06vFKbU5i_bRUaRwp0G_J8YvZK6zEudA8Zeiv6qdzDVMtZ-MObIgnVAH0\nhttps://mobile.twitter.com/thetimeusedtobe/status/1182292557595721730\n8) Raptor squad went undercover to target radical Hong Kong protesters These police disguised themselves as protesters. But in reality, they arrest vulnerable target like young girls and boys on the street in the name of action to arrest \"extremely violent rioters\". These target youngsters were bought into non-police vehicles by the undercover, then disappear without any trace and record.https://www.scmp.com/news/hong-kong/politics/article/3022457/elite-police-raptor-squad-went-undercover-target-radical\n9) Rumors of police kills peopleSince the occurrence of the 8.31 Prince Edward station incident, the \"police killing people\" rumors have been heard, and there have been many suspicious floating bodies, including Chen Yan-lin, a 15-year-old swimmer who had participated in the anti-Extradiction bill movement. The cause of death is unknown. Although the police denied twice, rumor is still spread. https://www.scmp.com/news/hong-kong/politics/article/3026439/hong-kong-protests-officials-attempt-sixth-time-debunk\n10) Inconsistent law enforcementhttps://www.hongkongfp.com/2019/07/26/hong-kong-police-breed-mistrust-uncertainty-selective-law-enforcement/\n11) Hong Kong becomes a police city In the name of law enforcement, the Hong Kong Police is allowed to extend their action of violence and brutality anywhere, anytime. Off-duty police officers were given telescopic batons and pepper spray for carrying and using when they were off duty.\n12) Spreading climate of fearThe Hong Kong Police Force has been accused of spreading a climate of fear. It has conducted hospital arrests, which had prompted protesters to become reluctant to go for treatment in public hospital, fearing they may get arrested by the police. Medical professionals may also be charged by the police for offering treatment to the wounded.The police banned protest and marches in various places in Hong Kong. The refusal to issue permits had eroded Hong Kong's freedom of demonstration. The police for rounding up various activists to frighten Hong Kong people into not protesting, even though these protests have been leaderless.Disregard the safety of the members of the public, police use unlawful force on peaceful or retreating protesters was usual. Hong Kong police can be seen in using excessive force against unarmed and non-resisting citizens and innocent commuters.\n13) Trash daily police press. Police lying to the public all the timeEveryday , it is anticipated that the Police Public Relations Bureau would be conducting another routine briefing at 4 pm Hong Kong time. It would not be surprising if they could find explanations, if not excuses, for the brutal force used and they would probably, if not certainly, put blames on the protesters for assembling illegally. It has also become a practice for the Police to condemn the protesters for violence while defending the police for whatever forces used during the operations, thereby damaging its own credibility.The Police has repeatedly denied accusations of tortures and sexual assault cases in detention center. The most notorious one was in San Uk Lang, which is proximate to the Chinese border. Victims had spoken both in public and online of the maltreatment that they have received and also what they have seen.In response, the police remained adamant that the accusations were false and demanded the victims to put away their mask and report to the police. One of the Deputy commissioner of police even issued a letter to fellow policemen, claiming that the accusation of “Black Cops” is non-existence and false.\n14) Without any conscious of damage cause by the unlawful and excessive use of force, Hong Kong police continues to consider its operation swell with unlimited power to enact laws.https://www.hongkongfp.com/2019/08/21/fall-hong-kong-police-force-new-branch-chinas-public-security-machinery/\nhttps://www.scmp.com/news/hong-kong/politics/article/3021258/beijing-stands-behind-hong-kong-police-state-media-while\nhttps://www.hongkongfp.com/2019/10/14/hong-kong-govt-considers-enlisting-staff-depts-become-police-special-constables-report/\nBeijing Stands Behind Hong Kong Police, Hong Kong People Need Help from the World\nEvery week, millions or thousands of citizens have come out to express their dissatisfaction towards the current political system and the performance of the government. Yet, the government chose to shift the responsibility of the political turmoil to the protesters and condemned violence selectively. They are turning a blind eye to the voice of the people, while pandering to the “white terror” and police brutality in Hong Kong.\nDespite the popular outcry from Hong Kong people from different sectors urging the Police Commissioner to issue an immediate official apology for its wilful attacks against citizens, people’s demand once again had not been heard. The reputation of HKPC is totally wrecked.\nAlthough we repeatedly demand the Independent Police Complaints Council (IPCC) to set up an independent inquiry to investigate HKPF’s misconduct and its gross abuse of power, once again the voice of the Hong Kong people is ignored. We seriously doubt of the independence, integrity and effectiveness of IPCC.\nMore peaceful rallies and protests will be ongoing held to openly condemn police excessive use of force as part of the ongoing anti-extradition bill protests. The Hong Kong people re-state their 5 major demands: \n(1) Independent inquiry on police brutality\n(2) Immediate release all protesters\n(3) Retract the characterisation of peaceful protests as “riots\" \n(4) Universal suffrage\n(5) Reform Hong Kong Police, disband the existing Police Forces\nWe are deeply worried that this unlawful use of force by the HKPF will persist. More unarmed peaceful protesters, mostly young people, will continued to be hurt and sustain a disproportionate level of injuries, some of which can be long term.\nWe seek for political settlement to end the violence that erupted following the anti-extradition bill in 2019. Human rights, political accountability and democracy should be introduced and put into the Hong Kong Police Ordinance. Police force is a major front on which these normative reforms were to take place. These reforms should seek to break the historical reliance of the political elite of Hong Kong/Mainland China on the police as an instrument of elite power and repression; to improve the account-and support the evolution of more a responsive, public-focused policing.\nWe implore the Permanent Court of Arbitration to take this crisis seriously and take immediate action to hold the HKPF to account. The Hong Kong government and the police force have completely failed to serve its people. It is only with the intervention of international organisations like the PCA, we will have a hope to restore order and peace on our streets in a place we call home.\nWe look forward to hearing from you soon.\nYour sincerely,Hong Kong Citizens\n(photo credit: Hong Kong Free Press)"],["topImageUrl","https://assets.change.org/photos/7/xd/mh/NmXdmHvvMnjVywm-800x450-noPad.jpg?1573414214"],["error",null]]}]}],["text","【請求荷蘭海牙常設仲裁法院調查香港暴警事件】\n這是具有法律效力的\n麻煩大家協助一下擴散跟連署！！\n\nhttps://www.change.org/p/request-permanent-court-of-arbitration-to-investigate-unlawful-use-of-force-of-hong-kong-police?utm_content=cl_sharecopy_17005953_en-GB%3Av5&recruiter=90670311&recruited_by_id=0040d850-5bec-43c4-8e81-c03a3953c79d&utm_source=share_petition&utm_medium=copylink&utm_campaign=psf_combo_share_message&utm_term=psf_combo_share_initial&share_bandit_exp=message-17005953-en-GB&share_bandit_var=v0"],["id","3mk40i15zdjrj"],["createdAt","2019-11-18T16:13:26.244Z"]]}],["reply",{"__iterable":"Map","data":[["id","2054gG4BV0DITCqGBW4O"],["user",{"__iterable":"Map","data":[["id","AVriAlOoyrDaTqlmmqhC"],["name","Lin"],["avatarUrl",null],["level",22]]}],["type","OPINIONATED"],["text","此為連署活動，提倡循法律途徑行動。"],["reference","https://hk.appledaily.com/local/20191109/IGGKFDHRZVUWH4OG5ZHIYIRXUI/"],["hyperlinks",{"__iterable":"List","data":[{"__iterable":"Map","data":[["title","練乙錚「眾籌到海牙仲裁警暴」｜日報｜要聞港聞｜ 20191109"],["url","https://hk.appledaily.com/local/20191109/IGGKFDHRZVUWH4OG5ZHIYIRXUI/"],["summary","【抗爭前路】【本報訊】逆權風暴已持續五個月，社會矛盾從未化解，政府越加打壓，市民對政府及警隊信心更低處未見低。時事評論員練乙錚認為社會已陷入死局，港人與中共的對立已到「漢賊不兩立」，他呼籲港人要繼續將香港所有矛盾呈現到全世界，包括區議會選舉。他又指既然政府怕開罪警隊而不作獨立調查，港人可眾籌到荷蘭海牙常設仲裁法院，仲裁港警行為，逼政府接受。記者：黃詠恩練乙錚指政府拒成立獨立調查委員會，建議可由海牙常設仲裁法院判斷警隊是否侵犯人權。資料圖片就本港局勢問題，練乙錚認為短期看不到化解的可能，政府不退讓，港人持續抗爭，面對膠着局面，他指既然政府怕惹警察不滿而不成立獨立調查委員會，港人大可眾籌將港警執法問題交由獨立的荷蘭海牙常設仲裁法院，判斷警隊是否真的侵犯人權，將整件事件國際化。香港2011年6月通過《香港仲裁條例》，讓法例能與國際仲裁制度接軌，目的是有意把香港發展成為國際仲裁中心，「就算仲裁結果對政府不利，喺國際上會畀政府好大壓力」，否則定會造成非常尷尬的局面。籲持續將香港問題國際化\n練乙錚指無論是國際仲裁或成立獨立調查委員會，都是文明世界處理政治事件的方法，但政府一一否定，證明特首林鄭月娥根本是一個野蠻政府，更加不肯問責落台，連犧牲一個半個管治班子都沒有，「林鄭自己唔肯負責，炒個律政司祭旗都好，令大家順氣啲」。他狠批林鄭留下一灘「蘇州屎」，不肯解決問題，令市民憤怒，商界對她亦非常反感。就高調支持林鄭的中央政府，練乙錚認為，中共處理新疆和西藏問題都是速戰速決，惟獨香港這戰場令中共最為頭痛：「以為香港地方細容易搞，其實唔係呀，邊疆所有棘手問題當中，香港呢場仗打最耐。」他認為中共要開竅才可以解決問題，始終在中美貿易戰下，香港是唯一未被打破的金蛋，仍獨具優勢。就今次抗爭，練乙錚稱想不到年輕人展現出的反抗力量如此強烈，認為中央若再強勢鎮壓，港人反抗心態只會越來越激進。除了眾籌到海牙仲裁外，他建議港人持續將香港的問題國際化，包括即將舉行的區議會選舉，「延遲選舉呢啲卑鄙手段完全可以做得出，所以喺呢個問題上唔好受佢挑釁，臨到選舉前全部靜落嚟」。他認為行動例如邀請外國記者到港或登廣告可將香港所有矛盾呈現到全世界，絕不能任由政府關門打狗。"],["topImageUrl","https://appledaily-hk-appledaily-prod.cdn.arcpublishing.com/resizer/RssV3zq32ubYPtOQRrIgwRSzky4=/d87urpdhi5rdo.cloudfront.net/11-08-2019/t_5d5b8662ff8e4b6f9d1d4cf7ccc077ea_name_1573234338_b8ae.jpg"],["error",null]]}]}],["createdAt","2019-11-18T21:42:59.601Z"]]}]]},{"__iterable":"Map","data":[["article",{"__iterable":"Map","data":[["replyCount",1],["references",{"__iterable":"List","data":[{"__iterable":"Map","data":[["type","LINE"]]}]}],["replyRequestCount",1],["user",{"__iterable":"Map","data":[["id","j4S8C_8SU6w_lKU8142Pd-hyVJRULJNF7mjehuE3om7Vc6i8c"],["name","滿足的✖東勢✖艾薇拉"],["avatarUrl",null],["level",0]]}],["hyperlinks",{"__iterable":"List","data":[{"__iterable":"Map","data":[["title","Diabetes drug is promising Alzheimer's treatment after 'significantly reversing' memory loss"],["url","http://www.independent.co.uk/news/health/alzheimers-diabetes-drug-memory-loss-reverse-disease-treatment-degenerative-a8133681.html"],["summary","A drug developed for type 2 diabetes significantly reverses memory loss and could have potential as a new treatment for Alzheimer’s disease and other neurodegenerative diseases, scientists say.\n \n\n \n \n \n \n \n\n\n\n\n\n\n\n\n \n \n \n\n\n\n\n\n \n \n \n \n \n The study, by UK and Chinese universities, is the first to look at a new combined diabetes drug and found improvements in several characteristic symptoms of Alzheimer’s.\n \n\n \n \n \n \n \n\n\n\n \n\n\n\n\n\n\n\n \n \n \n\n\n\n\n\n \n \n \n \n \n Lead researcher Professor Christian Holscher, from Lancaster University, said these “very promising outcomes” show multi-action drugs developed for type 2 diabetes “consistently show neurological protective effects”.\n \n\n \n \n \n \n \n\n\n\n\n \n \n \n\n\n\n\n\n \n \n \n\n\n\n\n\n \n \n \n \n \n Independent academics said a reduction in nerve-cell-killing protein molecules was particularly interesting and this was likely to be another avenue in the search for an elusive drug to combat dementia.\n \n\n \n \n \n \n \n\n\n\n\n\n \n\n\n\n\n\n \n \n \n\n\n\n\n\n \n \n \n \n \n He has previously reported optimistic findings from an older diabetes drug, liraglutide, and clinical trials in humans are currently under way.\n \n\n \n \n \n \n \n\n\n\n\n\n\n\n\n \n \n \n\n\n\n\n\n \n \n \n \n \n This latest study, published in the journal Brain Research, looked at a “triple action” treatment that combine three different drugs for type 2 diabetes, acting on biological pathways that could also have an impact on dementia.\n \n\n \n \n \n \n \n\n\n\n\n\n\n\n\n \n \n \n \n \n\n\n\n\n\n \n \n \n \n \n \n \n\n \n \n \n Type 2 diabetes is a known risk factor for Alzhemier’s disease and impaired production of insulin – the hormone that people with diabetes don’t produce sufficiently to control their blood sugar – is linked to brain degeneration.\n \n\n \n \n \n \n \n\n\n\n\n\n\n\n\n \n \n \n\n\n\n\n\n \n \n \n \n \n The identification of this link had a twofold benefit, according to charities.\n \n\n \n \n \n \n \n\n\n\n\n\n\n\n\n \n \n \n\n\n\n\n\n \n \n \n \n \n It opened up new research and drug development opportunities, such as this study.\n \n\n \n \n \n \n \n\n\n\n\n\n\n\n\n \n \n \n\n\n\n\n\n \n \n \n \n \n But it also means that by making lifestyle changes, like eating healthily and exercising, patients can avoid developing type-2 diabetes and lower their risk of Alzheimer’s disease as well.\n \n\n \n \n \n \n \n\n\n\n\n\n\n\n\n \n \n \n\n\n\n\n\n \n \n \n \n \n There are currently half a million people living with Alzheimer’s disease in the UK and more than five million people with diabetes, both conditions pose a major cost to the NHS, and health systems worldwide.\n \n\n \n \n \n \n \n\n\n\n\n\n\n\n\n \n \n \n\n\n\n\n\n \n \n \n \n \n \n\n \n\n \n \n \n The treatment in this study combined; GLP-1, GIP and glucagon, which, like insulin protect against neurological deterioration, and tested them in mice with genetic mutations that cause Alzheimer’s disease.\n \n\n \n \n \n \n \n\n\n\n\n\n\n\n\n \n \n \n\n\n\n\n\n \n \n \n \n \n After two months of daily injections the mice were shown to significantly improve their performance in a maze designed to test memory.\n \n\n \n \n \n \n \n\n\n\n\n\n\n\n\n \n \n \n \n \n\n \n\n\n\n\n\n \n \n \n \n \n Treated mice also had lower levels of proteins which clump together and form plaques in the brains of people with Alzheimer’s disease, inhibiting nerve cells’ ability to communicate and causing them to die.\n \n\n \n \n \n \n \n\n\n\n\n\n\n\n\n \n \n \n\n\n\n\n\n \n \n \n \n \n The study also found these mice lost nerve cells to the disease at a slower rate and had lower levels of nerve inflammation.\n \n\n \n \n \n \n \n\n\n\n\n\n\n\n\n \n \n \n\n\n\n\n\n \n \n \n \n \n It concludes that the triple treatment “holds clear promise of being developed into a new treatment for chronic neurodegenerative disorders such as Alzheimer’s disease”.\n \n\n \n \n \n \n \n\n\n\n\n\n\n\n\n \n \n \n \n \n\n \n\n\n\n\n\n \n \n \n \n \n \n \nThe seven Alzheimer's risk factors\n\n\n\n\n \n\n\n\n \n \n \n \n \n \n \n \n \n \n \n \n \n \n\n\n\n\n\n \n\n \n \n \n The Alzheimer’s Society, a charity which part-funded the latest study research, said that after 15 years without a new drug for Alzheimer’s disease the promising results from these crossover treatments, which are already known to be safe for humans, are likely to bring earliest benefits.\n \n\n \n \n \n \n \n\n\n\n\n\n\n\n\n \n \n \n\n\n\n\n\n \n \n \n \n \n Professor Holscher added that further tests to compare its benefits against other potential treatments, and its effects in humans were needed.\n \n\n \n \n \n \n \n\n\n\n\n\n\n\n\n \n \n \n\n\n\n\n\n \n \n \n \n \n But Professor John Hardy, a professor of neuroscience, at University College London who was not involved in the trial, said the study was “a first step, at best” towards a Alzheimer’s drug for humans.\n \n\n \n \n \n \n \n\n\n\n\n\n\n\n\n \n \n \n \n \n\n \n\n\n\n\n\n \n \n \n \n \n “The results showing less amyloid deposition of amyloid in mice treated with glucagon receptor stimulating drugs is interesting,” he said.\n \n\n \n \n \n \n \n \n “However, it should be noted that several other drugs have shown positive results in mice models of Alzheimer’s disease and then failed in human trials.”\n \n\n \n \n \n \n \n \n Dr David Reynolds, chief scientific officer at Alzheimer’s Research UK, echoed this, but added: “There is a long road between studies that show an effect in animals and treatments in the hands of patients, and scientists will only be able to realise the potential of promising findings like these if we continue to invest in research.”"],["topImageUrl","https://static.independent.co.uk/s3fs-public/thumbnails/image/2015/04/14/21/web-alzheimers-2-alamy.jpg"],["error",null]]},{"__iterable":"Map","data":[["title","Neuroprotective effects of a triple GLP-1/GIP/glucagon receptor agonist in the APP/PS1 transgenic mouse model of Alzheimer's disease"],["url","https://www.sciencedirect.com/science/article/pii/S0006899317304596"],["summary","AbstractType 2 diabetes mellitus (T2DM) is a risk factor for Alzheimer disease (AD). Previous studies have shown that the incretin hormones glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic polypeptide (GIP) that have anti-diabetic properties show very promising effects in animal models of AD. Glucagon (Gcg) is a hormone and growth-factor, and the Gcg receptor is expressed in the brain. Here we test the effects of a triple receptor agonist (TA), which activates GIP-1, GIP and glucagon receptors at the same time. In the present study, the effects of the TA were evaluated in the APP/PS1 transgenic mouse model of AD. The TA was injected once-daily (10 nmol/kg i.p.) for two months. The results showed that treatment with TA significantly reversed the memory deficit in the APP/PS1 mice in a spatial water maze test. Moreover, the drug reduced levels of the mitochondrial pro-apoptotic signaling molecule BAX, increased the anti-apoptotic signaling molecule Bcl-2 and enhanced the levels of BDNF, a key growth factor that protects synaptic function. Levels of synaptophysin were enhanced, demonstrating protection from synaptic loss that is observed in AD. Neurogenesis in the dentate gyrus was furthermore enhanced as shown in the increase of doublecortin positive cells. Furthermore, TA treatment reduced the total amount of β-amyloid, reduced neuroinflammation (activated microglia and astrocytes), and oxidative stress in the cortex and hippocampus. Thus, these findings show that novel TAs are a promising lead for the design of future treatment strategies in AD.Previous Next KeywordsInflammationGrowth factorBDNFBrainInsulinNeurodegeneration1. IntroductionAlzheimer’s disease (AD), is a progressive neurodegenerative disease, characterized clinically by progressive memory loss, cognitive decline, and aberrant behavior [1]. Currently, there is no treatment can improve this condition. Epidemiological studies have shown that type 2 diabetes mellitus (T2DM) is a risk factor for Alzheimer disease (AD) (Luchsinger et al., 2004, Ohara et al., 2011). The underlying mechanism is most likely that insulin signaling is impaired in the brains of AD patients (Moloney et al., 2010, Talbot et al., 2012). This motivated research in drugs that have shown good effects in treating diabetes to investigate if they could be helpful in treating AD as well. Previous studies have shown that the incretin hormones glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic polypeptide (GIP) can play a neuroprotective role in the brain and show very promising effects in animal models of AD (Duffy and Holscher, 2013, Faivre and Holscher, 2013b, Li et al., 2010, McClean et al., 2011, McClean and Holscher, 2014). Glucagon (Gcg) is a hormone and growth-factor, and the Gcg receptor is expressed in the brain (Hoosein and Gurd, 1984, Mayo et al., 2003). Activation of the receptor can activate neuronal activity and cellular Ca2+ signaling and may have neuroprotective properties (Ayush et al., 2015). Here we test the effects of a triple receptor agonist (TA), which activates GIP-1, GIP and glucagon receptors at the same time. This novel drug has been developed as a potential treatment for diabetes (Finan et al., 2015). No studies have been published that test the neuroprotective effects of this novel drug. We therefore tested this promising TA in the APPSWE/PS1dE9 mouse model of AD. This mouse model recapitulates some of the hallmarks of AD, such as memory loss, synaptic loss, reduction of synaptic plasticity, reduction of neurogenesis in the dentate gyrus, chronic inflammation in the brain, and formation of amyloid plaques in the brain (Duffy and Holscher, 2013, Goto et al., 2008, Hamilton et al., 2011, Hamilton and Holscher, 2012, McClean et al., 2011). We tested the effects of TA in this transgenic mouse model and evaluated the neuroprotective effects. We analyzed memory formation, hippocampal neurogenesis, the expression levels of Brain Derived Neurotrophic Factor (BDNF) (Blurton-Jones et al., 2009), mitochondrial apoptosis signaling proteins, neuroinflammation and oxidative stress levels.2. Results2.1. TA-treatment improved the learning and memory impairment of APP/PS1 mice in the water mazeA two-way ANOVA found a significant difference between groups (p < 0.0001) and over time (p < 0.001). In the Morris water test, it was observed that the control mice, APP/PS1 mice and TA-treated APP/PS1 mice had a similar escape latency (P > 0.05). On day 5, the average escape distance in searching for the hidden platform in the APP/PS1 mice was significantly longer than that in the control mice (P < 0.05). Compared to the APP/PS1 group, the TA-treated APP/PS1 group’s average escape distance was decreased (P < 0.01). There was no difference in swimming speed among these three groups in training days (P > 0.05).Then on day 6, the platform was removed and the probe trail test was conducted. A one way ANOVA analysis showed an overall difference between groups. The APP/PS1 mice spent less time in the target quadrant (P < 0.01) and had fewer times passing through the original position of the hidden platform (P < 0.01), compared with control mice. In the TA-treated APP/PS1 group, the time spent in target quadrant (P < 0.05) and the frequencies to cross the platform significantly increased (P < 0.05), compared to the APP/PS1 group Fig. 1.Download high-res image (302KB)Download full-size imageFig. 1. The novel TA improved the learning ability of APP/PS1 mice in the spatial water maze task. (A) The escape latency and swimming speed in the training trails. (B) The percentage of time in target quadrant and crossing platform times in the probe trials. The circle indicates the position of the platform during acquisition (C) Representative swimming tracks of the probe trial. * = P < .05, ** = P < .01 compared to the control group, ## = P < .01, # = P < .05 compared to the APP/PS1 group; Two-way ANOVA with Bonferroni repeated measures post hoc tests.2.2. TA-treatment ameliorated amyloid plaques in the brains of APP/PS1 miceIn the immunohistochemical analysis, we analyzed the amount of amyloid plaques present in the brains of APP-PS1 mice. A one-way ANOVA found an overall difference on the amyloid deposits in the cortex (F = 31.10, P < 0.0001) and the hippocampus (F = 30.38, P < 0.0001) of mice between groups. The Aβ1-42 depositions in the cortex and hippocampus of the APP/PS1 mice were significantly higher than that in the control mice (P < 0.001), while TA-treatment group significantly decreased the deposition (P < 0.001) compared to the APP/PS1 group. N = 6 per group. See Fig. 2.Download high-res image (211KB)Download full-size imageFig. 2. TA treatment reduced amyloid deposits in the cortex and hippocampus of APP/PS1 mice. A, D: control group; B, E: APP/PS1 group; C, F: APP/PS1+TA group. Scale bar in image D: 100 μm. *** = P < .001 compared to the control group; ### = P < .001 compared to the APP/PS1 group. One-way ANOVA with Bonferroni repeated measures post hoc tests. N = 6 per group.2.3. TA-treatment reduced the chronic inflammation response in the cortex and hippocampus of the APP/PS1 miceIn the immunohistochemical analysis, an anti-GFAP antibody was used as an indication of activated astrocytes. A one-way ANOVA found an overall difference on the GFAP expression in the cortex (F = 77.03, P < 0.0001) and the hippocampus (F = 86.51, P < 0.0001) of mice between groups. The levels of GFAP in the cortex and hippocampus of the APP/PS1 mice were significantly higher than that in the control mice (P < 0.001), while TA-treatment group significantly decreased (P < 0.001) compared to the APP/PS1 group. N = 6 per group. See Fig. 3.Download high-res image (362KB)Download full-size imageFig. 3. TA-treatment reduced the number of GFAP positive cells in the cortex and hippocampus of APP/PS1 mice. A, D: control group; B, E: APP/PS1 group; C, F: APP/PS1+TA. Scale bar in image D: 50μm. A higher magnification image has been added to show stained astroglia. *** = P < .001 compared with the control group; ### = P < .001 compared with the APP/PS1 group. One-way ANOVA with Bonferroni repeated measures post hoc tests. N = 6 per group.In the immunohistochemical analysis, an anti-IBA1 antibody was used as an indication of microglia activation. A one-way ANOVA found an overall difference on the IBA-1 expression in the cortex (F = 28.71, P < 0.0001) and the hippocampus (F = 66.42, P < 0.0001) of mice between groups. The levels of IBA-1 in the cortex and hippocampus of the APP/PS1 mice were significantly higher than that in the control mice (P < 0.001), while TA-treatment group significantly decreased (P < 0.001, P < 0.01) compared to the APP/PS1 group. N = 6 per group. See Fig. 4.Download high-res image (347KB)Download full-size imageFig. 4. TA treatment reversed the increase of IBA-1 positive cells in the cortex and hippocampus of APP/PS1 mice. A, D: control group; B, E: APP/PS1 group; C, F: APP/PS1+TA group. Scale bar in image D: 100 μm. A higher magnification image has been added to show stained microglia. *** = P < .001 compared to the control group; ## = P < .01, ### = P < .001 compared to the APP/PS1 group. One-way ANOVA with Bonferroni repeated measures post hoc tests. N = 6 per group.2.4. TA-treatment suppressed oxidative stress in the cortex and hippocampus of APP /PS1 miceIn the immunohistochemical analysis, 4-HNE was measured as an indicator of lipid peroxidation. A one-way ANOVA found an overall difference on the 4-HNE expression in the cortex (F = 60.60, P < 0.0001) and the hippocampus (F = 35.22, P < 0.0001) of mice between groups. The expressions of 4-HNE in the cortex and hippocampus of the APP/PS1 mice were significantly higher than that in the control mice (P < 0.001), while TA-treatment group significantly decreased (P < 0.001) compared to the APP/PS1 group. N = 6 per group, see Fig. 5a.Download high-res image (397KB)Download full-size imageFig. 5. Oxidative stress was reduced by the TA. 5A. TA-treatment reduced the increase of 4-HNE levels in the cortex and hippocampus in APP/PS1 mice. A, D: control group; B, E: APP/PS1 group; C, F: APP/PS1+TA group. Scale bar in image D: 100 μm. *** = P < .001 compared with controls; ###=P < .001 compared to the APP/PS1 group. 5B. TA treatment reduced the increase of 8-OHdG levels in the cortex and hippocampus of APP/PS1 mice. A, D: control group; B, E: APP/PS1 group; C, F: APP/PS1+TA group. Scale bar in image D: 100 μm. *** = P < .001 compared to the control group; ## = P < .01, ### = P < .001 compared with the APP/PS1 group. One-way ANOVA with Bonferroni repeated measures ***post hoc tests. N = 6 per group.In the immunohistochemical analysis, 8-OHdG was measured as an indicator of DNA oxidation. A one-way ANOVN found an overall difference on the 8-OHdG expression in the cortex (F = 38.72, P < 0.0001) and the hippocampus (F = 56.60, P < 0.0001) of mice between groups. The expressions of 8-OHdG in the cortex and hippocampus of the APP/PS1 mice were significantly higher than that in the control mice (P < 0.001), while TA-treatment group significantly decreased (P < 0.01, P < 0.001) compared to the APP/PS1 group. N = 6 per group, see Fig. 5b.2.5. TA-treatment increased immature neurons in the dentate gyrus of APP/PS1 miceIn the immunohistochemical analysis, numbers of neurogenesis, assessed by doublecortin (DCX). A one-way ANOVN found an overall difference on the DCX positive cell in the dentate gyrus (F = 43.61, P < 0.0001) of mice between groups. The numbers of DCX were much reduced in the APP/PS1 mice (P < 0.001) compared to the control mice. While TA-treatment partially prevented the reduction (P < 0.001) compared to the APP/PS1 mice. N = 6 per group. See Fig. 6.Download high-res image (169KB)Download full-size imageFig. 6. TA-treatment enhanced neurogenesis and increased numbers of DCX positive cell numbers in the dentate gyrus of APP/PS1 mice. A: control group; B: APP/PS1 group; C: APP/PS1+TA group. Scale bar in image A: 100 μm. *** = P < .001 compared with the control group; ### = P < .001 compared with the APP/PS1 group. One-way ANOVA with Bonferroni repeated measures post hoc tests. N = 6 per group.2.6. TA-treatment reversed the decrease of BDNF levels in the hippocampus of APP/PS1 miceBDNF, a key neurotrophic factor in the brain, was detected by Western blot analysis. A one-way ANOVA found an overall difference between groups (F = 20.01, P < 0.001). The expression of BDNF in hippocampus was reduced in APP/PS1 group (P < 0.001) compared to the control group. TA-treatment reversed the decreased (P < 0.05) compared to the APP/PS1 group. N = 6 per group. See Fig. 7a.Download high-res image (304KB)Download full-size imageFig. 7. A) TA-treatment increased levels of BDNF in the hippocampus of APP/PS1 mice. *** = P < .001compared with the control group; # = P < .05 compared with the APP/PS1 group. N = 6 per group. B) TA-treatment increased levels of synaptophysin in the hippocampus of APP/PS1 mice. *** = P < .001 compared with the control group; ## = P < .01 compared with the APP/PS1 group. N = 6 per group. C) TA-treatment increased the ratio of BAX/Bcl-2 in the hippocampus of APP/PS1 mice. *** = P < .001 compared with the control group; ### = P < .001 compared with the APP/PS1 group. One-way ANOVA with Bonferroni repeated measures post hoc tests. N = 6 per group.2.7. TA-treatment reversed the reduction of synaptophysin levelsSynaptophysin (SYN), a synaptic markers, was detected by Western blot analysis. A one-way ANOVA found an overall difference between groups (F = 20.46, P < 0.001). The level of SYN in hippocampus was decreased in the APP/PS1 group (P < 0.001) compared to the control group. While TA-treatment prevent the reduction (P < 0.01) compared to the APP/PS1 group. N = 6 per group. See Fig. 7b.2.8. TA-treatment showed potential anti-apoptotic effects by decreasing the BAX/Bcl-2 ratio in APP/PS1 miceThe expression of apoptotic proteins, including Bcl-2 (that suppress apoptosis) and BAX (that promotes apoptosis), was detected by Western blot analysis. A one-way ANOVA found an overall difference between groups (F = 77.64, P < 0.001). The ratio of BAX/Bcl-2 in hippocampus was increased (P < 0.001) compared to the control group. While TA-treatment partly decreased the ratio of BAX/Bcl-2 (P < 0.001) compared to the APP/PS1 group. N = 6 per group. See Fig. 7c.3. DiscussionThe results demonstrate for the first time that the novel GLP-1/GIP/Gcg receptor agonist has clear neuroprotective effects in the APP/PS1 mouse model of AD. Memory formation in the spatial water maze task was improved by the drug, and the amyloid plaque load in the cortex and hippocampus was reduced. This result confirms our previous findings that single GLP-1 or GIP analogues can protect from memory loss in this mouse model of AD (Faivre and Holscher, 2013a, Faivre and Holscher, 2013b, McClean et al., 2011, McClean and Holscher, 2014). The chronic inflammation response as seen in the activation of microglia and astrocytes is much reduced by the novel TA, similar to single GLP-1 or GIP receptor agonists (Duffy and Holscher, 2013, Ji et al., 2016a, McClean et al., 2011). Levels of the peroxidized lipid 4-HNE that also acts as a pro-apoptotic signal in mitochondria have been reduced by the TA. 4-HNE is one of the markers of membrane lipid peroxidation induced by cytotoxic radicals such as OH. 4-HNE has cytotoxic, mutagenic and genotoxic properties and 4-HNE is reported to stimulate apoptosis via caspase 3 activation and by inducing mitochondrial damage as shown in cytochrome c release (Ji et al., 2001). The levels of the mitochondrial apoptosis signal BAX (Kim et al., 2005) were also reduced. Importantly, the anti-apoptotic signaling molecule B-cell lymphoma 2 (Bcl-2) which is active in functioning mitochondria that have normal inner membrane voltage potentials (Kavitha et al., 2013) is upregulated by the TA, adding to the neuroprotective effect. The level of chronic oxidative stress as measure by the levels of the oxidized nucleotide 8-Hydroxiguanosine (8-OHdG) was also reduced by the drug. Neurogenesis in the dentate gyrus was normalized. Importantly, the levels of the key growth factor BDNF were normalized in the brain. BDNF has shown clear neuroprotective effects in a range of animal models of disease (Allen et al., 2013, Benedetti et al., 2014, Blurton-Jones et al., 2009, Nagahara et al., 2013). An important factor of BNDF activity is that it maintains synaptic activity and protects synapses from stressors (Kuipers and Bramham, 2006). The observation that the synaptic marker synaptophysin was expressed at normal levels after TA treatment also indicates that synaptic numbers and function was improved in the AD mouse. We have shown previously that GLP-1 and GIP analogues protect synapses from amyloid induced stress and keeps them functional (Faivre and Holscher, 2013b, Gault and Holscher, 2008a, Gault and Holscher, 2008b, McClean and Holscher, 2014).Recently, multiple receptor agonists have been developed to treat type II diabetes (Finan et al., 2013, Finan et al., 2015). We have tested novel dual GLP-1/GIP receptor agonists that showed good neuroprotective effects that are superior to single GLP-1 analogues (Cao et al., 2016, Jalewa et al., 2017, Ji et al., 2016b, Yuan et al., 2017). The novel dual agonist DA3-CH was found to be superior to liraglutide in a mouse model of Parkinson’s disease (Yuan et al., 2017), demonstrating that the strategy of activating more than one incretin receptor type offers an advantage over single receptor agonists. Here, we show that a novel triple receptor agonist also shows promise as a potential treatment for AD, but further dose–response tests and direct comparisons with other drugs have to be conducted in order to evaluate if this new drug is superior to previous ones.GLP-1, GIP and glucagon are members of the growth factor family (Hölscher, 2016). Their respective receptors are classic 7-membrane spanning domain, G-protein coupled receptors that activate cAMP synthesis (Campbell and Drucker, 2013, Doyle and Egan, 2007, Holst et al., 2011). The activation of the second messenger cascade cAMP-PKA-CREB induces a number of cytoprotective processes, including the normalization of energy utilization, cell repair and growth-factor related gene expression (Holscher, 2014, Jalewa et al., 2016, Perry and Greig, 2004, Sharma et al., 2013). Importantly, GLP-1 and GIP also have anti-inflammatory properties. Astroglia and microglia express the receptors, and activating these reduce the inflammation response (Duffy and Holscher, 2013, Jalewa et al., 2017, Parthsarathy and Holscher, 2013, Spielman et al., 2017). Chronic inflammation is a main driver of neurodegenerative disorders, and a reduction of the inflammation in the brain shows protective effects (Akiyama et al., 2000, Clark et al., 2012, Lee et al., 2010).Based on the impressive preclinical data that demonstrate neuroprotection of such drugs, clinical trials have started that investigate the neuroprotective effects of the GLP-1 receptor agonists exendin-4 (Byetta, Bydureon®) and liraglutide (Victoza®) in patients with AD or with Parkinson disease (PD). A pilot study testing the GLP-1 analogue liraglutide in AD patients showed promising results. In a double blind, placebo controlled trial, AD patients showed no deterioration in brain activity as measured in the 18FDG-PET scan after 12 months of drug treatment. In contrast, the placebo group showed clear reductions in cortical activity as expected for this progressive neurodegenerative disease (Gejl et al., 2016). A phase II clinical trial is currently ongoing (Hölscher, 2016). A pilot trial testing exendin-4 in PD patients has shown good effects (NCT01174810). This open-label study tested the effects of exendin-4 in 45 patients. There were clear improvements in the motor coordination as shown in the MDS-UPDRS part 3 assessment, and cognition was improved as measured by the Mattis DRS-2 test battery that had been designed to evaluate cognitive impairments in PD patients (Aviles-Olmos et al., 2013). Patients were re-tested 12 months later, and the differences in motor performance and cognitive scores were still visible (Aviles-Olmos et al., 2014). A phase II double-blind, placebo controlled clinical trial that tested the once-weekly formulation of exendin-4, Bydureon®, confirmed the results of the pilot study. The drug group did not deteriorate in the MDS-UPDRS part 3 assessment after 48 weeks of treatment, and after 12 weeks of a wash-out period, the disease-modifying effect was still visible. In contrast, the placebo group deteriorated over time as would be expected in this disease (Athauda et al., 2017). In addition, a phase II trial testing liraglutide in PD patients has started, testing patients in a double blind, placebo controlled trial for one year (NCT02953665). A further clinical trial testing the GLP-1 receptor agonist Lixisenatide in PD patients is in planning.These very promising preclinical and the first clinical outcomes demonstrate the efficacy of these novel multiple receptor agonists that originally were developed to treat type II diabetes, but have shown consistent neuroprotective effects in several studies. In the present study, the novel TA improved learning and memory, reduce amyloid plaque load, chronic inflammation and oxidative stress, as well as mitochondrial stress signaling, and improved BDNF release and synaptic markers while normalising neurogenesis in the dentate gyrus. The novel triple GLP-1/GIP/Gcg receptor agonist holds clear promise of being developed into a new treatment for chronic neurodegenerative disorders such as Alzheimer’s disease.4. Materials and methods4.1. Peptide and chemicalsThe triple GLP-1/GIP/Gcg used in this study was synthesized by China Peptides Co, Ltd. (Shanghai, China) to 95% purity. The identity and purity of the peptide was confirmed by reversed-phase HPLC and characterized using matrix assisted laser resorption/ionization time of flight (MALDI–TOF) mass spectrometry. The peptide was stored in dry form and dissolved in double-distilled water containing 0.9% NaCl2 before experiments. A Bi-cinchoninicacid (BCA) protein assay kit was purchased from Apply gen Technologies Inc. (Beijing, China). 3, 3-diaminobenzidine (DAB) was purchased from ZSGB-BIO Co. (Beijing, China). The primary antibodies recognizing Aβ1-42, IBA-1, BDNF, 4-Hydroxynonenal (4-HNE), 8-Hydroxyguanosine (8-OHdG), synaptophysin (SYN) and β-actin were obtained from Abcam (Cambridge, UK). The primary antibody GFAP, Bcl-2, and BAX were obtained from Boster Biotechnology (Wuhan, China). The primary antibody doublecortin (DCX) buy from Santa Cruz Biotechnology. HRP-labeled goat anti-rabbit immunoglobulin was obtained from Abcam (Cambridge, UK). The biotinylated goat anti-rabbit IgG, biotinylated rabbit anti-goat IgG, avidinbiotin peroxidase complex reagent was purchased from Boster Biotechnology (Wuhan, China). All other reagents used in the research were of analytical grade and of the highest quality commercially available. The peptide sequence was as follows (Finan et al., 2015):HXQGTFTSDKSKYLDERAAQDFVQWLLDGGPSSGAPPPSX = aminoisobutyric acid, K = γE-C16 acylation (with a C16 fatty acid).4.2. Animals and treatmentsAPP/PS1 transgenic mice were purchased from the Academy of Military Medical Sciences (AMMS, China), were maintained in a constant temperature (22 ± 1 °C) and humidity (relative 30%) under a 12 h light/dark cycle in animal house. As well as age-matched non-AD mice, which are wild-type animals as control. When animals were 6 months of age, TA was dissolved in saline. The dose was a once-daily injection of 10 nmol/kg body weight ip. for two months. Injections were given in the morning (8:00 am). Control mice received equal volumes of saline solution. All experimental procedures involving animals were approved by the Institutional Animal Care Committee of Shanxi Medical University and conform to the guidelines of National Institute of Health (NIH) guideline (NIH publication No. 85-23. Revised 1985). They were divided into three different groups (n = 12). (i): control group treated with saline only; (ii): APP/PS1 group treated with saline only; (iii): APP/PS1 group treated with TA. At the end of drug treatments, measurements of behavioral change, neuronal damage, oxidative stress and inflammatory markers were conducted.4.3. Morris water maze testThe water maze test is a standard method for assessing learning and memory in animals. The water maze (WM) task was performed 5 days after two months intraperitoneal injection. The apparatus was a circular, white painted pool (150 cm in diameter and 60 cm in height) which was filled to a depth of 40 cm with warm water at a temperature 23 ± 2 °C to avoid hypothermia, and was surrounded by a white curtain around the pool. The pool was divided into four quadrants with four starting locations, north (N), east (E), south (S) and west (W), the starting quadrant was randomly chosen. A perspex platform (diameter, 10 cm) was located in a specific location during the acquisition trials and submerged approximately 1.0 cm below the surface of the water. A video camera was mounted directly above the pool to record the movement and the trail of the mice. During the acquisition phase, each mouse was subjected to four consecutive training trials on each day. The mouse was gently placed in the water between the quadrants, and allowed 60 s to search the hidden platform. If it failed to find the platform within 60 s, it was guided to the platform and allowed to remain there for 10 s. Training continued for 5 days. On the sixth day, the hidden platform was removed and each mouse was allowed to swim in the pool for 60 s. The mean time spent in all four quadrants and the target area for searching the hidden platform was recorded.4.4. Brain tissue preparationHalf of the animals were decapitated after anesthetization with ethyl carbamate, hippocampus tissues were immediately isolated and stored at −80 °C for further investigations. The second half of each group was used for immunohistochemistry analysis. Animals were perfused intracardially with saline and cold 4% paraformaldehyde. Brains were removed and post-fixed in 4% paraformaldehyde for 24 h.4.5. ImmunohistochemistryFor DAB staining, fixed tissues were embedded in paraffin. Coronal sections (5 μm thick) of the brains were cut on a Leica microtome for immunohistochemistry. Paraffin was removed from the tissue sections with xylene, and the sections were rehydrated in graded ethanol solutions. Endogenous peroxidase activity was blocked with 3% H2O2 for 10 min at room temperature. Antigen retrieval was performed by heating in 10 mmol/L citrate buffer (pH 6.0) for 10 min. After blocked with 5% BSA, sections were incubated with the primary antibody for doublecortin (goat anti-DCX; 1:50), GFAP (rabbit anti-GFAP; 1:100), IBA-1 (goat anti-IBA1; 1:200), 4-Hydroxynonenal antibody (goat anti-4 HNE; 1:200) and anti-8 Hydroxyguanosine antibody (goat anti-8 OHdG; 1:200) at 37 °C for 2 h. For the Anti-8 OHdG histology, enzymatic antigen retrieval with 10 μm/ml proteinase K for 40 min at 37 °C was performed. To detect amyloid deposits, the primary antibody was Aβ1-42 (rabbit anti-Aβ1-42; 1:100), brain sections were pre-incubated with 70% formic acid and incubated at 4 °C for 24 h. They were rinsed in PBS and incubated with biotinylated goat anti-rabbit IgG or biotinylated rabbit anti-goat IgG at 37 °C for 0.5 h and incubated with the avidin-biotin peroxidase complex reagent at 37 °C for 0.5 h. Samples were stained in DAB solution in the presence of H2O2. All stained sections were viewed and photographed under a Zeiss light microscope, and images were captured by Motic Images Plus2.0ML software. The numbers of DCX positive neurons, the areas of Aβ1-42, GFAP, IBA-1, 4-HNE and 8-OHdG positive cells were determined using Image-pro plus6.0 software. The software counts objects that are defined in size and staining threshold by the user. This automated process avoided user bias.4.6. Western blotThe hippocampal tissues from mice was homogenized in ice cold RIPA buffer (containing 1% Triton X-100, 0.1% SDS, 1% deoxycholate) and PMSF (phenyl-methylsulfonyl fluoride). The homogenate was centrifuged 12,000 rpm/min for 5 min at 4 °C before taking the supernatant. Tissue containing 60 μg of protein was separated on 10–12% gradient Bis–Tris gel with molecular marker and electrophoresed in running buffer at 120 mV for 90 min followed by transfer to the PVDF membrane. Following protein transfer, the membrane was washed in TBST (tris buffered saline with 0.05% Tween-20, pH 8) and blocked in 5% skim milk for 1 h. Then incubated overnight at 4 °C with rabbit anti-synaptophysin (SYN) (1:10,000), rabbit anti-Bcl-2 (1:400), rabbit anti-BAX (1:400), rabbit anti-BDNF (1:5000), after three washes in TBS further incubation for 2 h at room temperature with the HRP-labeled goat anti-rabbit immunoglobulin (1:5000). In some cases, the blots were stripped and reprobed with a rabbit anti-β-actin (1:5000) to ensure equal sample loading. The bound antibodies were then visualized by ECL-enhanced chemiluminescence (Beyotime Institute of Biotechnology, Shanghai, China) according to the manufacturer's instructions. Western blot images were captured with a chemiluminescent imaging system (Sage Creation, Beijing, China). All bands were quantified using the image system of Alpha View SA.4.7. Statistical analysisThe data were presented as the mean ± standard error of mean (S.E.M.) and analyzed by one-way or two way ANOVA followed by Bonferroni’s multiple comparison test using the software Prism 5 (Graph Pad Software Inc, La Jolla, CA, USA). Statistical significance is defined as P < 0.05.ContributorsJT, WL and YL conducted the experiments, LL and CH conceived the experiment and wrote the manuscript.AcknowledgementsThis work was funded by the Ministry of Human Resources and Social Security, Shanxi Province [(2010)255], by a Shanxi Scholarship Council of China, and supported by a grant from the Alzheimer Society UK. The authors declare no conflict of interestAppendix A. Supplementary dataReferencesAkiyama et al., 2000H. Akiyama, S. Barger, S. Barnum, B. Bradt, J. Bauer, G.M. Cole, N.R. Cooper, P. Eikelenboom, M. Emmerling, B.L. Fiebich, C.E. Finch, S. Frautschy, W.S. Griffin, H. Hampel, M. Hull, G. Landreth, L. Lue, R. Mrak, I.R. Mackenzie, P.L. McGeer, M.K. O'Banion, J. Pachter, G. Pasinetti, C. Plata-Salaman, J. Rogers, R. Rydel, Y. Shen, W. Streit, R. Strohmeyer, I. Tooyoma, F.L. Van Muiswinkel, R. Veerhuis, D. Walker, S. Webster, B. Wegrzyniak, G. Wenk, T. Wyss-CorayInflammation and Alzheimer’s diseaseNeurobiol. Aging, 21 (2000), pp. 383-421Allen et al., 2013S.J. Allen, J.J. Watson, D.K. Shoemark, N.U. Barua, N.K. PatelGDNF, NGF and BDNF as therapeutic options for neurodegenerationPharmacol. Ther., 138 (2013), pp. 155-175Athauda et al., 2017D. Athauda, K. Maclagan, S.S. Skene, M. Bajwa-Joseph, D. Letchford, K. Chowdhury, S. Hibbert, N. Budnik, L. Zampedri, J. Dickson, Y. Li, I. Aviles-Olmos, T.T. Warner, P. Limousin, A.J. Lees, N.H. Greig, S. Tebbs, T. FoltynieExenatide once weekly versus placebo in Parkinson’s disease: a randomised, double-blind, placebo-controlled trialAviles-Olmos et al., 2013I. Aviles-Olmos, J. Dickson, Z. Kefalopoulou, A. Djamshidian, P. Ell, T. Soderlund, P. Whitton, R. Wyse, T. Isaacs, A. Lees, P. Limousin, T. FoltynieExenatide and the treatment of patients with Parkinson’s diseaseJ. Clin. Invest., 123 (2013), pp. 2730-2736Aviles-Olmos et al., 2014I. Aviles-Olmos, J. Dickson, Z. Kefalopoulou, A. Djamshidian, J. Kahan, P.E. Fmedsci, P. Whitton, R. Wyse, T. Isaacs, A. Lees, P. Limousin, T. FoltynieMotor and cognitive advantages persist 12 months after exenatide exposure in Parkinson’s diseaseJ. Parkinson’s Dis., 4 (2014), pp. 337-344Ayush et al., 2015E.A. Ayush, Y. Iwasaki, S. Iwamoto, H. Nakabayashi, M. Kakei, T. YadaGlucagon directly interacts with vagal afferent nodose ganglion neurons to induce Ca(2+) signaling via glucagon receptorsBiochem. Biophys. Res. Commun., 456 (2015), pp. 727-732Benedetti et al., 2014E. Benedetti, B. D'Angelo, L. Cristiano, E. Di Giacomo, F. Fanelli, S. Moreno, F. Cecconi, A. Fidoamore, A. Antonosante, R. Falcone, R. Ippoliti, A. Giordano, A. CiminiInvolvement of peroxisome proliferator-activated receptor beta/delta (PPAR beta/delta) in BDNF signaling during aging and in Alzheimer disease: possible role of 4-hydroxynonenal (4-HNE)Cell Cycle, 13 (2014), pp. 1335-1344Blurton-Jones et al., 2009M. Blurton-Jones, M. Kitazawa, H. Martinez-Coria, N.A. Castello, F.J. Muller, J.F. Loring, T.R. Yamasaki, W.W. Poon, K.N. Green, F.M. LaferlaNeural stem cells improve cognition via BDNF in a transgenic model of Alzheimer diseaseProc. Natl. Acad. Sci. U.S.A., 106 (2009), pp. 13594-13599Campbell and Drucker, 2013J.E. Campbell, D.J. DruckerPharmacology, physiology, and mechanisms of incretin hormone actionCell Metab., 17 (2013), pp. 819-837Cao et al., 2016L. Cao, D. Li, P. Feng, L. Li, G. Xue, G. Li, C. HölscherA novel dual GLP-1 and GIP incretin receptor agonist is neuroprotective in a mouse model of Parkinson’s disease by reducing chronic inflammation in the brainNeuroreport, 37 (2016), pp. 384-391Clark et al., 2012I. Clark, C. Atwood, R. Bowen, G. Paz-Filho, B. VisselTumor necrosis factor-induced cerebral insulin resistance in Alzheimer's disease links numerous treatment rationalesPharmacol. Rev., 64 (2012), pp. 1004-1026Doyle and Egan, 2007M.E. Doyle, J.M. EganMechanisms of action of glucagon-like peptide 1 in the pancreasPharmacol. Ther., 113 (2007), pp. 546-593Duffy and Holscher, 2013A.M. Duffy, C. HolscherThe incretin analogue D-Ala(2)GIP reduces plaque load, astrogliosis and oxidative stress in an APP/PS1 mouse model of Alzheimer’s diseaseNeuroscience, 228 (2013), pp. 294-300Faivre and Holscher, 2013aE. Faivre, C. HolscherD-Ala2GIP facilitated synaptic plasticity and reduces plaque load in aged wild type mice and in an Alzheimer's disease mouse modelJ. Alzheimers Dis., 35 (2013), pp. 267-283Faivre and Holscher, 2013bE. Faivre, C. HolscherNeuroprotective effects of D-Ala2GIP on Alzheimer’s disease biomarkers in an APP/PS1 mouse modelAlzheimers Res. Ther., 5 (2013), pp. 20-28Finan et al., 2013B. Finan, T. Ma, N. Ottaway, T.D. Muller, K.M. Habegger, K.M. Heppner, H. Kirchner, J. Holland, J. Hembree, C. Raver, S.H. Lockie, D.L. Smiley, V. Gelfanov, B. Yang, S. Hofmann, D. Bruemmer, D.J. Drucker, P.T. Pfluger, D. Perez-Tilve, J. Gidda, L. Vignati, L. Zhang, J.B. Hauptman, M. Lau, M. Brecheisen, S. Uhles, W. Riboulet, E. Hainaut, E. Sebokova, K. Conde-Knape, A. Konkar, R.D. DiMarchi, M.H. TschopUnimolecular dual incretins maximize metabolic benefits in rodents, monkeys, and humansSci. Transl. Med., 5 (2013), p. 209ra151Finan et al., 2015B. Finan, B. Yang, N. Ottaway, D.L. Smiley, T. Ma, C. Clemmensen, J. Chabenne, L. Zhang, K.M. Habegger, K. Fischer, J.E. Campbell, D. Sandoval, R.J. Seeley, K. Bleicher, S. Uhles, W. Riboulet, J. Funk, C. Hertel, S. Belli, E. Sebokova, K. Conde-Knape, A. Konkar, D.J. Drucker, V. Gelfanov, P.T. Pfluger, T.D. Muller, D. Perez-Tilve, R.D. DiMarchi, M.H. TschopA rationally designed monomeric peptide triagonist corrects obesity and diabetes in rodentsNat. Med., 21 (2015), pp. 27-36Gault and Holscher, 2008aV.A. Gault, C. HolscherGLP-1 agonists facilitate hippocampal LTP and reverse the impairment of LTP induced by beta-amyloidEur. J. Pharmacol., 587 (2008), pp. 112-117Gault and Holscher, 2008bV.A. Gault, C. HolscherProtease-resistant glucose-dependent insulinotropic polypeptide agonists facilitate hippocampal LTP and reverse the impairment of LTP induced by beta-amyloidJ. Neurophysiol., 99 (2008), pp. 1590-1595Gejl et al., 2016M. Gejl, A. Gjedde, L. Egefjord, A. Møller, S.B. Hansen, K. Vang, A.B. Rodell, H. Braendgaard, H. Gottrup, A. Schacht, N. Møller, B. Brock, J. RungbyIn Alzheimer’s disease, six-month treatment with glp-1 analogue prevents decline of brain glucose metabolism: randomized, placebo-controlled double-blind clinical trialFront. Aging Neurosci., 8 (2016), pp. 1-10Goto et al., 2008Y. Goto, T. Niidome, H. Hongo, A. Akaike, T. Kihara, H. SugimotoImpaired muscarinic regulation of excitatory synaptic transmission in the APPswe/PS1dE9 mouse model of Alzheimer's diseaseEur. J. Pharmacol., 583 (2008), pp. 84-91Hamilton et al., 2011A. Hamilton, S. Patterson, D. Porter, V.A. Gault, C. HolscherNovel GLP-1 mimetics developed to treat type 2 diabetes promote progenitor cell proliferation in the brainJ. Neurosci. Res., 89 (2011), pp. 481-489Hamilton and Holscher, 2012A. Hamilton, C. HolscherThe effect of ageing on neurogenesis and oxidative stress in the APP(swe)/PS1(deltaE9) mouse model of Alzheimer's diseaseBrain Res., 1449 (2012), pp. 83-93Holscher, 2014C. HolscherInsulin, incretins and other growth factors as potential novel treatments for Alzheimer's and Parkinson's diseasesBiochem. Soc. Trans., 42 (2014), pp. 593-599Hölscher, 2016C. HölscherGLP-1 and GIP analogues as novel treatments for Alzheimer’s and Parkinson’s diseaseCardiovasc. Endocrinol., 5 (2016), pp. 93-98Holst et al., 2011J.J. Holst, R. Burcelin, E. NathansonNeuroprotective properties of GLP-1: theoretical and practical applicationsCurr. Med. Res. Opin., 27 (2011), pp. 547-558Hoosein and Gurd, 1984N.M. Hoosein, R.S. GurdIdentification of glucagon receptors in rat brainProc. Natl. Acad. Sci. U.S.A., 81 (1984), pp. 4368-4372Jalewa et al., 2016J. Jalewa, M.K. Sharma, C. HolscherNovel incretin analogues improve autophagy and protect from mitochondrial stress induced by rotenone in SH-SY5Y cellsJ. Neurochem., 139 (2016), pp. 55-67Jalewa et al., 2017J. Jalewa, M. Sharma, S. Gengler, C. HölscherA novel GLP-1/GIP dual receptor agonist protects from 6-OHDA lesion in a rat model of Parkinson’s diseaseNeuropharmacology, 117 (2017), pp. 238-248Ji et al., 2001C. Ji, V. Amarnath, J.A. Pietenpol, L.J. Marnett4-hydroxynonenal induces apoptosis via caspase-3 activation and cytochrome c releaseChem. Res. Toxicol., 14 (2001), pp. 1090-1096Ji et al., 2016aC. Ji, G.F. Xue, G. Li, D. Li, C. HolscherNeuroprotective effects of glucose-dependent insulinotropic polypeptide in Alzheimer's diseaseRev. Neurosci., 27 (2016), pp. 61-70Ji et al., 2016bC. Ji, G.F. Xue, C. Lijun, P. Feng, D. Li, L. Li, G. Li, C. HolscherA novel dual GLP-1 and GIP receptor agonist is neuroprotective in the MPTP mouse model of Parkinson's disease by increasing expression of BNDFBrain Res., 1634 (2016), pp. 1-11Kavitha et al., 2013M. Kavitha, J. Nataraj, M.M. Essa, M.A. Memon, T. ManivasagamMangiferin attenuates MPTP induced dopaminergic neurodegeneration and improves motor impairment, redox balance and Bcl-2/Bax expression in experimental Parkinson's disease miceChem. Biol. Interact., 206 (2013), pp. 239-247Kim et al., 2005S.J. Kim, K. Winter, C. Nian, M. Tsuneoka, Y. Koda, C.H. McIntoshGlucose-dependent insulinotropic polypeptide (GIP) stimulation of pancreatic beta-cell survival is dependent upon phosphatidylinositol 3-kinase (PI3K)/protein kinase B (PKB) signaling, inactivation of the forkhead transcription factor Foxo1, and down-regulation of bax expressionJ. Biol. Chem., 280 (2005), pp. 22297-22307Kuipers and Bramham, 2006S.D. Kuipers, C.R. BramhamBrain-derived neurotrophic factor mechanisms and function in adult synaptic plasticity: new insights and implications for therapyCurr. Opin. Drug Discov. Dev., 9 (2006), pp. 580-586Lee et al., 2010Y.J. Lee, S.B. Han, S.Y. Nam, K.W. Oh, J.T. HongInflammation and Alzheimer's diseaseArch. Pharm. Res., 33 (2010), pp. 1539-1556Li et al., 2010Y. Li, K. Duffy, M. Ottinger, B. Ray, J. Bailey, H. Holloway, D. Tweedie, T. Perry, M. Mattson, D. Kapogiannis, K. Sambamurti, D. Lahiri, N. GreigGLP-1 receptor stimulation reduces amyloid-beta peptide accumulation and cytotoxicity in cellular and animal models of Alzheimer's diseaseJ. Alzheimers Dis., 19 (2010), pp. 1205-1219Luchsinger et al., 2004J.A. Luchsinger, M.X. Tang, S. Shea, R. MayeuxHyperinsulinemia and risk of Alzheimer diseaseNeurology, 63 (2004), pp. 1187-1192Mayo et al., 2003K.E. Mayo, L.J. Miller, D. Bataille, S. Dalle, B. Goke, B. Thorens, D.J. DruckerInternational Union of Pharmacology. XXXV. The glucagon receptor familyPharmacol. Rev., 55 (2003), pp. 167-194McClean et al., 2011P. McClean, V. Parthsarathy, E. Faivre, C. HölscherThe diabetes drug liraglutide prevents degenerative processes in a mouse model of Alzheimer's diseaseJ. Neurosci., 31 (2011), pp. 6587-6594McClean and Holscher, 2014P.L. McClean, C. HolscherLixisenatide, a drug developed to treat type 2 diabetes, shows neuroprotective effects in a mouse model of Alzheimer's diseaseNeuropharmacology, 86C (2014), pp. 241-258Moloney et al., 2010A.M. Moloney, R.J. Griffin, S. Timmons, R. O'Connor, R. Ravid, C. O'NeillDefects in IGF-1 receptor, insulin receptor and IRS-1/2 in Alzheimer's disease indicate possible resistance to IGF-1 and insulin signallingNeurobiol. Aging, 31 (2010), pp. 224-243Nagahara et al., 2013A.H. Nagahara, M. Mateling, I. Kovacs, L. Wang, S. Eggert, E. Rockenstein, E.H. Koo, E. Masliah, M.H. TuszynskiEarly BDNF treatment ameliorates cell loss in the entorhinal cortex of APP transgenic miceJ. Neurosci., 33 (2013), pp. 15596-15602Ohara et al., 2011T. Ohara, Y. Doi, T. Ninomiya, Y. Hirakawa, J. Hata, T. Iwaki, S. Kanba, Y. KiyoharaGlucose tolerance status and risk of dementia in the community: the Hisayama studyNeurology, 77 (2011), pp. 1126-1134Parthsarathy and Holscher, 2013V. Parthsarathy, C. HolscherThe type 2 diabetes drug liraglutide reduces chronic inflammation induced by irradiation in the mouse brainEur. J. Pharmacol., 700 (2013), pp. 42-50Perry and Greig, 2004T.A. Perry, N.H. GreigA new Alzheimer's disease interventive strategy: GLP-1Curr. Drug Targets, 5 (2004), pp. 565-571Sharma et al., 2013M. Sharma, J. Jalewa, C. HolscherNeuroprotective and anti-apoptotic effects of liraglutide on SH-SY5Y cells exposed to Methylglyoxal stressJ. Neurochem., 128 (2013), pp. 459-471Spielman et al., 2017L.J. Spielman, D.L. Gibson, A. KlegerisIncretin hormones regulate microglia oxidative stress, survival and expression of trophic factorsEur. J. Cell Biol., 96 (2017), pp. 240-253Talbot et al., 2012K. Talbot, H.Y. Wang, H. Kazi, L.Y. Han, K.P. Bakshi, A. Stucky, R.L. Fuino, K.R. Kawaguchi, A.J. Samoyedny, R.S. Wilson, Z. Arvanitakis, J.A. Schneider, B.A. Wolf, D.A. Bennett, J.Q. Trojanowski, S.E. ArnoldDemonstrated brain insulin resistance in Alzheimer's disease patients is associated with IGF-1 resistance, IRS-1 dysregulation, and cognitive declineJ. Clin. Invest., 122 (2012), pp. 1316-1338Yuan et al., 2017Z. Yuan, L. Li, P. Feng, G. Xue, C. Ji, G. Li, C. HölscherA novel GLP-1/GIP dual agonist is more effective than liraglutide in reducing inflammation and enhancing GDNF release in the MPTP mouse model of Parkinson’s disease© 2017 Elsevier B.V. All rights reserved."],["topImageUrl","https://ars.els-cdn.com/content/image/1-s2.0-S0006899317X0022X-cov150h.gif"],["error",null]]}]}],["text","阿茲海默症(Alzheimer’s disease)疾病對於病人及家屬皆會帶來很大的影響，然而至目前為止，疾病的發生機轉及可能的治療方式卻仍在研究階段。《Cell Reports》日前刊登中國山西醫科大學(Shanxi Medical University)的最新發表，研究團隊表示，原先發展用來治療糖尿病的特定藥物，有機會反轉阿茲海默症的記憶喪失症狀。第二型糖尿病已知為阿茲海默症的風險因子之一，其病程會影響到胰島素的分泌，而在過去的研究中亦曾發現，這個影響人體血糖調節的荷爾蒙，其分泌不足可能與大腦的退化情形有關。本篇研究中，作者發現結合第二型糖尿病的特定藥物，可能改善數項阿茲海默症的獨特病徵。作者表示，這項研究發現，有助於未來發展阿茲海默症及其它神經退化疾病的新治療方法。\n \n新聞出處：Alzheimer's treatment: Diabetes drug holds promise for fighting disease after 'significantly reversing' memory loss《Independent》\nhttp://www.independent.co.uk/news/health/alzheimers-diabetes-drug-memory-loss-reverse-disease-treatment-degenerative-a8133681.html\n研究刊登在《Cell Reports》第1678卷第64-74頁\nhttps://www.sciencedirect.com/science/article/pii/S0006899317304596\n "],["id","AWEET-BFyCdS-nWhunGr"],["createdAt","2018-01-17T13:29:59.107Z"]]}],["reply",{"__iterable":"Map","data":[["id","AWEElGNbyCdS-nWhunHT"],["user",{"__iterable":"Map","data":[["id","AVriAlOoyrDaTqlmmqhC"],["name","Lin"],["avatarUrl",null],["level",22]]}],["type","RUMOR"],["text","美國布朗大學沃倫阿爾伯特醫學院（The Warren Alpert Medical School of Brown University）的蘇珊教授（Suzanne de la Monte），後續發現阿茲海默症患者不一定有第一型或第二型糖尿病，但腦部也會出現血糖代謝異常的狀況，因此認為阿茲海默症和糖尿病有高度相似性，因此學者提出阿茲海默症為第三型糖尿病的說法。"],["reference","http://www.cofit.me/229093913535506/4688052\n飲食與生活習慣的改善，才是預防阿茲海默症的根本"],["hyperlinks",{"__iterable":"List","data":[{"__iterable":"Map","data":[["title","This area is password protected [401]"],["url","http://www.cofit.me/229093913535506/4688052"],["summary","This area is password protected"],["topImageUrl",""],["error",null]]}]}],["createdAt","2018-01-17T14:44:49.119Z"]]}]]},{"__iterable":"Map","data":[["article",{"__iterable":"Map","data":[["replyCount",1],["references",{"__iterable":"List","data":[{"__iterable":"Map","data":[["type","LINE"]]}]}],["replyRequestCount",2],["user",{"__iterable":"Map","data":[["id","j4S8C_YlgNaHHUAmuAJvBJ9fT06nPwQIphzAJEbrZaWWhlh_8"],["name","發起者的頭屋維拉"],["avatarUrl",null],["level",0]]}],["hyperlinks",{"__iterable":"List","data":[{"__iterable":"Map","data":[["title","Respiratory virus shedding in exhaled breath and efficacy of face masks"],["url","https://www.nature.com/articles/s41591-020-0843-2?fbclid=IwAR3EzP6Xm2bXH4bDHx9i9oOYxqHIraWbuRbZcGzM6fmgQz7iIl7ufHrkN-A"],["summary","AbstractWe identified seasonal human coronaviruses, influenza viruses and rhinoviruses in exhaled breath and coughs of children and adults with acute respiratory illness. Surgical face masks significantly reduced detection of influenza virus RNA in respiratory droplets and coronavirus RNA in aerosols, with a trend toward reduced detection of coronavirus RNA in respiratory droplets. Our results indicate that surgical face masks could prevent transmission of human coronaviruses and influenza viruses from symptomatic individuals.\n \n \n \n\n \n \n MainRespiratory virus infections cause a broad and overlapping spectrum of symptoms collectively referred to as acute respiratory virus illnesses (ARIs) or more commonly the ‘common cold’. Although mostly mild, these ARIs can sometimes cause severe disease and death1. These viruses spread between humans through direct or indirect contact, respiratory droplets (including larger droplets that fall rapidly near the source as well as coarse aerosols with aerodynamic diameter >5 µm) and fine-particle aerosols (droplets and droplet nuclei with aerodynamic diameter ≤5 µm)2,3. Although hand hygiene and use of face masks, primarily targeting contact and respiratory droplet transmission, have been suggested as important mitigation strategies against influenza virus transmission4, little is known about the relative importance of these modes in the transmission of other common respiratory viruses2,3,5. Uncertainties similarly apply to the modes of transmission of COVID-19 (refs. 6,7).Some health authorities recommend that masks be worn by ill individuals to prevent onward transmission (source control)4,8. Surgical face masks were originally introduced to protect patients from wound infection and contamination from surgeons (the wearer) during surgical procedures, and were later adopted to protect healthcare workers against acquiring infection from their patients. However, most of the existing evidence on the filtering efficacy of face masks and respirators comes from in vitro experiments with nonbiological particles9,10, which may not be generalizable to infectious respiratory virus droplets. There is little information on the efficacy of face masks in filtering respiratory viruses and reducing viral release from an individual with respiratory infections8, and most research has focused on influenza11,12.Here we aimed to explore the importance of respiratory droplet and aerosol routes of transmission with a particular focus on coronaviruses, influenza viruses and rhinoviruses, by quantifying the amount of respiratory virus in exhaled breath of participants with medically attended ARIs and determining the potential efficacy of surgical face masks to prevent respiratory virus transmission.ResultsWe screened 3,363 individuals in two study phases, ultimately enrolling 246 individuals who provided exhaled breath samples (Extended Data Fig. 1). Among these 246 participants, 122 (50%) participants were randomized to not wearing a face mask during the first exhaled breath collection and 124 (50%) participants were randomized to wearing a face mask. Overall, 49 (20%) voluntarily provided a second exhaled breath collection of the alternate type.Infections by at least one respiratory virus were confirmed by reverse transcription PCR (RT–PCR) in 123 of 246 (50%) participants. Of these 123 participants, 111 (90%) were infected by human (seasonal) coronavirus (n = 17), influenza virus (n = 43) or rhinovirus (n = 54) (Extended Data Figs. 1 and 2), including one participant co-infected by both coronavirus and influenza virus and another two participants co-infected by both rhinovirus and influenza virus. These 111 participants were the focus of our analyses.There were some minor differences in characteristics of the 111 participants with the different viruses (Table 1a). Overall, 24% of participants had a measured fever ≥37.8 °C, with patients with influenza more than twice as likely than patients infected with coronavirus and rhinovirus to have a measured fever. Coronavirus-infected participants coughed the most with an average of 17 (s.d. = 30) coughs during the 30-min exhaled breath collection. The profiles of the participants randomized to with-mask versus without-mask groups were similar (Supplementary Table 1).Table 1a Characteristics of individuals with symptomatic coronavirus, influenza virus or rhinovirus infectionWe tested viral shedding (in terms of viral copies per sample) in nasal swabs, throat swabs, respiratory droplet samples and aerosol samples and compared the latter two between samples collected with or without a face mask (Fig. 1). On average, viral shedding was higher in nasal swabs than in throat swabs for each of coronavirus (median 8.1 log10 virus copies per sample versus 3.9), influenza virus (6.7 versus 4.0) and rhinovirus (6.8 versus 3.3), respectively. Viral RNA was identified from respiratory droplets and aerosols for all three viruses, including 30%, 26% and 28% of respiratory droplets and 40%, 35% and 56% of aerosols collected while not wearing a face mask, from coronavirus, influenza virus and rhinovirus-infected participants, respectively (Table 1b). In particular for coronavirus, we identified OC43 and HKU1 from both respiratory droplets and aerosols, but only identified NL63 from aerosols and not from respiratory droplets (Supplementary Table 2 and Extended Data Fig. 3).Fig. 1: Efficacy of surgical face masks in reducing respiratory virus shedding in respiratory droplets and aerosols of symptomatic individuals with coronavirus, influenza virus or rhinovirus infection.a–c, Virus copies per sample collected in nasal swab (red), throat swab (blue) and respiratory droplets collected for 30min while not wearing (dark green) or wearing (light green) a surgical face mask, and aerosols collected for 30min while not wearing (brown) or wearing (orange) a face mask, collected from individuals with acute respiratory symptoms who were positive for coronavirus (a), influenza virus (b) and rhinovirus (c), as determined by RT–PCR in any samples. P values for mask intervention as predictor of log10 virus copies per sample in an unadjusted univariate Tobit regression model which allowed for censoring at the lower limit of detection of the RT–PCR assay are shown, with significant differences in bold. For nasal swabs and throat swabs, all infected individuals were included (coronavirus, n=17; influenza virus, n=43; rhinovirus, n=54). For respiratory droplets and aerosols, numbers of infected individuals who provided exhaled breath samples while not wearing or wearing a surgical face mask, respectively were: coronavirus (n=10 and 11), influenza virus (n=23 and 28) and rhinovirus (n=36 and 32). A subset of participants provided exhaled breath samples for both mask interventions (coronavirus, n=4; influenza virus, n=8; rhinovirus, n=14). The box plots indicate the median with the interquartile range (lower and upper hinge) and ±1.5×interquartile range from the first and third quartile (lower and upper whiskers).Table 1b Efficacy of surgical face masks in reducing respiratory virus frequency of detection and viral shedding in respiratory droplets and aerosols of symptomatic individuals with coronavirus, influenza virus or rhinovirus infectionWe detected coronavirus in respiratory droplets and aerosols in 3 of 10 (30%) and 4 of 10 (40%) of the samples collected without face masks, respectively, but did not detect any virus in respiratory droplets or aerosols collected from participants wearing face masks, this difference was significant in aerosols and showed a trend toward reduced detection in respiratory droplets (Table 1b). For influenza virus, we detected virus in 6 of 23 (26%) and 8 of 23 (35%) of the respiratory droplet and aerosol samples collected without face masks, respectively. There was a significant reduction by wearing face masks to 1 of 27 (4%) in detection of influenza virus in respiratory droplets, but no significant reduction in detection in aerosols (Table 1b). Moreover, among the eight participants who had influenza virus detected by RT–PCR from without-mask aerosols, five were tested by viral culture and four were culture-positive. Among the six participants who had influenza virus detected by RT–PCR from with-mask aerosols, four were tested by viral culture and two were culture-positive. For rhinovirus, there were no significant differences between detection of virus with or without face masks, both in respiratory droplets and in aerosols (Table 1b). Conclusions were similar in comparisons of viral shedding (Table 1b). In addition, we found a significant reduction in viral shedding (Supplementary Table 2) in respiratory droplets for OC43 (Extended Data Fig. 4) and influenza B virus (Extended Data Fig. 5) and in aerosols for NL63 (Extended Data Fig. 4).We identified correlations between viral loads in different samples (Extended Data Figs. 6–8) and some evidence of declines in viral shedding by time since onset for influenza virus but not for coronavirus or rhinovirus (Extended Data Fig. 9). In univariable analyses of factors associated with detection of respiratory viruses in various sample types, we did not identify significant association in viral shedding with days since symptom onset (Supplementary Table 3) for respiratory droplets or aerosols (Supplementary Tables 4–6).A subset of participants (72 of 246, 29%) did not cough at all during at least one exhaled breath collection, including 37 of 147 (25%) during the without-mask and 42 of 148 (28%) during the with-mask breath collection. In the subset for coronavirus (n = 4), we did not detect any virus in respiratory droplets or aerosols from any participants. In the subset for influenza virus (n = 9), we detected virus in aerosols but not respiratory droplets from one participant. In the subset for rhinovirus (n = 17), we detected virus in respiratory droplets from three participants, and we detected virus in aerosols in five participants.DiscussionOur results indicate that aerosol transmission is a potential mode of transmission for coronaviruses as well as influenza viruses and rhinoviruses. Published studies detected respiratory viruses13,14 such as influenza12,15 and rhinovirus16 from exhaled breath, and the detection of SARS-CoV17 and MERS-CoV18 from air samples (without size fractionation) collected from hospitals treating patients with severe acute respiratory syndrome and Middle East respiratory syndrome, but ours demonstrates detection of human seasonal coronaviruses in exhaled breath, including the detection of OC43 and HKU1 from respiratory droplets and NL63, OC43 and HKU1 from aerosols.Our findings indicate that surgical masks can efficaciously reduce the emission of influenza virus particles into the environment in respiratory droplets, but not in aerosols12. Both the previous and current study used a bioaerosol collecting device, the Gesundheit-II (G-II)12,15,19, to capture exhaled breath particles and differentiated them into two size fractions, where exhaled breath coarse particles >5 μm (respiratory droplets) were collected by impaction with a 5-μm slit inertial Teflon impactor and the remaining fine particles ≤5 μm (aerosols) were collected by condensation in buffer. We also demonstrated the efficacy of surgical masks to reduce coronavirus detection and viral copies in large respiratory droplets and in aerosols (Table 1b). This has important implications for control of COVID-19, suggesting that surgical face masks could be used by ill people to reduce onward transmission.Among the samples collected without a face mask, we found that the majority of participants with influenza virus and coronavirus infection did not shed detectable virus in respiratory droplets or aerosols, whereas for rhinovirus we detected virus in aerosols in 19 of 34 (56%) participants (compared to 4 of 10 (40%) for influenza and 8 of 23 (35%) for coronavirus). For those who did shed virus in respiratory droplets and aerosols, viral load in both tended to be low (Fig. 1). Given the high collection efficiency of the G-II (ref. 19) and given that each exhaled breath collection was conducted for 30 min, this might imply that prolonged close contact would be required for transmission to occur, even if transmission was primarily via aerosols, as has been described for rhinovirus colds20. Our results also indicate that there could be considerable heterogeneity in contagiousness of individuals with coronavirus and influenza virus infections.The major limitation of our study was the large proportion of participants with undetectable viral shedding in exhaled breath for each of the viruses studied. We could have increased the sampling duration beyond 30 min to increase the viral shedding being captured, at the cost of acceptability in some participants. An alternative approach would be to invite participants to perform forced coughs during exhaled breath collection12. However, it was the aim of our present study to focus on recovering respiratory virus in exhaled breath in a real-life situation and we expected that some individuals during an acute respiratory illness would not cough much or at all. Indeed, we identified virus RNA in a small number of participants who did not cough at all during the 30-min exhaled breath collection, which would suggest droplet and aerosol routes of transmission are possible from individuals with no obvious signs or symptoms. Another limitation is that we did not confirm the infectivity of coronavirus or rhinovirus detected in exhaled breath. While the G-II was designed to preserve viability of viruses in aerosols, and in the present study we were able to identify infectious influenza virus in aerosols, we did not attempt to culture coronavirus or rhinovirus from the corresponding aerosol samples.MethodsStudy designParticipants were recruited year-round from March 2013 through May 2016 in a general outpatient clinic of a private hospital in Hong Kong. As routine practice, clinic staff screened all individuals attending the clinics for respiratory and any other symptoms regardless of the purpose of the visit at triage. Study staff then approached immediately those who reported at least one of the following symptoms of ARI for further screening: fever ≥37.8 °C, cough, sore throat, runny nose, headache, myalgia and phlegm. Individuals who reported ≥2 ARI symptoms, within 3 d of illness onset and ≥11 years of age were eligible to participate. After explaining the study to and obtaining informed consent from the participants, a rapid influenza diagnostic test, the Sofia Influenza A + B Fluorescent Immunoassay Analyzer (cat. no. 20218, Quidel), was used to identify influenza A or B virus infection as an incentive to participate. All participants provided a nasal swab for the rapid test and an additional nasal swab and a separate throat swab for subsequent virologic confirmation at the laboratory. All participants also completed a questionnaire to record basic information including age, sex, symptom severity, medication, medical conditions and smoking history. In the first phase of the study from March 2013 to February 2014 (‘Influenza Study’), the result of the rapid test was used to determine eligibility for further participation in the study and exhaled breath collection, whereas in the second phase of the study from March 2014 to May 2016 (‘Respiratory Virus Study’), the rapid test did not affect eligibility. Eligible participants were then invited to provide an exhaled breath sample for 30 min in the same clinic visit.Before exhaled breath collection, each participant was randomly allocated in a 1:1 ratio to either wearing a surgical face mask (cat. no. 62356, Kimberly-Clark) or not during the collection. To mimic the real-life situation, under observation by the study staff, participants were asked to attach the surgical mask themselves, but instruction on how to wear the mask properly was given when the participant wore the mask incorrectly. Participants were instructed to breathe as normal during the collection, but (natural) coughing was allowed and the number of coughs was recorded by study staff. Participants were then invited to provide a second exhaled breath sample of the alternate type (for example if the participant was first assigned to wearing a mask they would then provide a second sample without a mask), but most participants did not agree to stay for a second measurement because of time constraints. Participants were compensated for each 30-min exhaled breath collection with a supermarket coupon worth approximately US$30 and all participants were gifted a tympanic thermometer worth approximately US$20.Ethical approvalWritten informed consent was obtained from all participants ≥18 years of age and written informed consent was obtained from parents or legal guardians of participants 11–17 years of age in addition to their own written informed consent. The study protocol was approved by the Institutional Review Board of The University of Hong Kong and the Clinical and Research Ethics Committee of Hong Kong Baptist Hospital.Collection of swabs and exhaled breath particlesNasal swabs and throat swabs were collected separately, placed in virus transport medium, stored and transported to the laboratory at 2–8 °C and the virus transport medium was aliquoted and stored at −70 °C until further analysis. Exhaled breath particles were captured and differentiated into two size fractions, the coarse fraction containing particles with aerodynamic diameter >5 μm (referred to here as ‘respiratory droplets’), which included droplets up to approximately 100 µm in diameter and the fine fraction with particles ≤5 μm (referred to here as ‘aerosols’) by the G-II bioaerosol collecting device12,15,19. In the G-II device, exhaled breath coarse particles >5 μm were collected by a 5-μm slit inertial Teflon impactor and the remaining fine particles ≤5 μm were condensed and collected into approximately 170 ml of 0.1% BSA/PBS. Both the impactor and the condensate were stored and transported to the laboratory at 2–8 °C. The virus on the impactor was recovered into 1 ml and the condensate was concentrated into 2 ml of 0.1% BSA/PBS, aliquoted and stored at −70 °C until further analysis. In a validation study, the G-II was able to recover over 85% of fine particles >0.05 µm in size and had comparable collection efficiency of influenza virus as the SKC BioSampler19.Laboratory testingSamples collected from the two studies were tested at the same time. Nasal swab samples were first tested by a diagnostic-use viral panel, xTAG Respiratory Viral Panel (Abbott Molecular) to qualitatively detect 12 common respiratory viruses and subtypes including coronaviruses (NL63, OC43, 229E and HKU1), influenza A (nonspecific, H1 and H3) and B viruses, respiratory syncytial virus, parainfluenza virus (types 1–4), adenovirus, human metapneumovirus and enterovirus/rhinovirus. After one or more of the candidate respiratory viruses was detected by the viral panel from the nasal swab, all the samples from the same participant (nasal swab, throat swab, respiratory droplets and aerosols) were then tested with RT–PCR specific for the candidate virus(es) for determination of virus concentration in the samples. Infectious influenza virus was identified by viral culture using MDCK cells as described previously21, whereas viral culture was not performed for coronavirus and rhinovirus.Statistical analysesThe primary outcome of the study was virus generation rate in tidal breathing of participants infected by different respiratory viruses and the efficacy of face masks in preventing virus dissemination in exhaled breath, separately considering the respiratory droplets and aerosols. The secondary outcomes were correlation between viral shedding in nose swabs, throat swabs, respiratory droplets and aerosols and factors affecting viral shedding in respiratory droplets and aerosols.We identified three groups of respiratory viruses with the highest frequency of infection as identified by RT–PCR, namely coronavirus (including NL63, OC43, HKU1 and 229E), influenza virus and rhinovirus, for further statistical analyses. We defined viral shedding as log10 virus copies per sample and plotted viral shedding in each sample (nasal swab, throat swab, respiratory droplets and aerosols); the latter two were stratified by mask intervention. As a proxy for the efficacy of face masks in preventing transmission of respiratory viruses via respiratory droplet and aerosol routes, we compared the respiratory virus viral shedding in respiratory droplet and aerosol samples between participants wearing face masks or not, by comparing the frequency of detection with a two-sided Fisher’s exact test and by comparing viral load (defined as log10 virus copies per sample) by an unadjusted univariate Tobit regression model, which allowed for censoring at the lower limit of detection of the RT–PCR assay. We also used the unadjusted univariate Tobit regression to investigate factors affecting viral shedding in respiratory droplets and aerosols without mask use, for example age, days since symptom onset, previous influenza vaccination, current medication and number of coughs during exhaled breath collection. We investigated correlations between viral shedding in nasal swab, throat swab, respiratory droplets and aerosols with scatter-plots and calculated the Spearman’s rank correlation coefficient between any two types of samples. We imputed 0.3 log10 virus copies ml−1 for undetectable values before transformation to log10 virus copies per sample. All analyses were conducted with R v.3.6.0 (ref. 22) and the VGAM package v.1.1.1 (ref. 23).Reporting SummaryFurther information on research design is available in the Nature Research Reporting Summary linked to this article.\n \n\n Data availabilityAnonymized raw data and R syntax to reproduce all the analyses, figures, tables and supplementary tables in the published article are available at: https://doi.org/10.5061/dryad.w9ghx3fkt.References1.Nichols, W. G., Peck Campbell, A. J. & Boeckh, M. Respiratory viruses other than influenza virus: impact and therapeutic advances. Clin. Microbiol. Rev. 21, 274–290 (2008).2.Shiu, E. Y. C., Leung, N. H. L. & Cowling, B. J. Controversy around airborne versus droplet transmission of respiratory viruses: implication for infection prevention. Curr. Opin. Infect. Dis. 32, 372–379 (2019).3.Tellier, R., Li, Y., Cowling, B. J. & Tang, J. W. Recognition of aerosol transmission of infectious agents: a commentary. BMC Infect. Dis. 19, 101 (2019).4.Xiao, J. et al. Nonpharmaceutical measures for pandemic influenza in nonhealthcare settings-personal protective and environmental measures. Emerg. Infect. Dis. https://doi.org/10.3201/eid2605.190994 (2020).5.Kutter, J. S., Spronken, M. I., Fraaij, P. L., Fouchier, R. A. M. & Herfst, S. Transmission routes of respiratory viruses among humans. Curr. Opin. Virol. 28, 142–151 (2018).6.Cowling, B. J. & Leung, G. M. Epidemiological research priorities for public health control of the ongoing global novel coronavirus (2019-nCoV) outbreak. Euro Surveill. https://doi.org/10.2807/1560-7917.ES.2020.25.6.2000110 (2020).7.Han, Q., Lin, Q., Ni, Z. & You, L. Uncertainties about the transmission routes of 2019 novel coronavirus. Influenza Other Respir. Viruses https://doi.org/10.1111/irv.12735 (2020).8.MacIntyre, C. R. & Chughtai, A. A. Facemasks for the prevention of infection in healthcare and community settings. BMJ 350, h694 (2015).9.Ha’eri, G. B. & Wiley, A. M. The efficacy of standard surgical face masks: an investigation using “tracer particles”. Clin. Orthop. Relat. Res. 148, 160–162 (1980).10.Patel, R. B., Skaria, S. D., Mansour, M. M. & Smaldone, G. C. Respiratory source control using a surgical mask: an in vitro study. J. Occup. Environ. Hyg. 13, 569–576 (2016).11.Johnson, D. F., Druce, J. D., Birch, C. & Grayson, M. L. A quantitative assessment of the efficacy of surgical and N95 masks to filter influenza virus in patients with acute influenza infection. Clin. Infect. Dis. 49, 275–277 (2009).12.Milton, D. K., Fabian, M. P., Cowling, B. J., Grantham, M. L. & McDevitt, J. J. Influenza virus aerosols in human exhaled breath: particle size, culturability, and effect of surgical masks. PLoS Pathog. 9, e1003205 (2013).13.Huynh, K. N., Oliver, B. G., Stelzer, S., Rawlinson, W. D. & Tovey, E. R. A new method for sampling and detection of exhaled respiratory virus aerosols. Clin. Infect. Dis. 46, 93–95 (2008).14.Stelzer-Braid, S. et al. Exhalation of respiratory viruses by breathing, coughing and talking. J. Med. Virol. 81, 1674–1679 (2009).15.Yan, J. et al. Infectious virus in exhaled breath of symptomatic seasonal influenza cases from a college community. Proc. Natl Acad. Sci. USA 115, 1081–1086 (2018).16.Tovey, E. R. et al. Rhinoviruses significantly affect day-to-day respiratory symptoms of children with asthma. J. Allergy Clin. Immunol. 135, 663–669 (2015).17.Booth, T. F. et al. Detection of airborne severe acute respiratory syndrome (SARS) coronavirus and environmental contamination in SARS outbreak units. J. Infect. Dis. 191, 1472–1477 (2005).18.Kim, S. H. et al. Extensive viable Middle East respiratory syndrome (MERS) coronavirus contamination in air and surrounding environment in MERS isolation wards. Clin. Infect. Dis. 63, 363–369 (2016).19.McDevitt, J. J. et al. Development and performance evaluation of an exhaled-breath bioaerosol collector for influenza virus. Aerosol Sci. Technol. 47, 444–451 (2013).20.Jennings, L. C. & Dick, E. C. Transmission and control of rhinovirus colds. Eur. J. Epidemiol. 3, 327–335 (1987).21.Chan, K. H., Peiris, J. S., Lim, W., Nicholls, J. M. & Chiu, S. S. Comparison of nasopharyngeal flocked swabs and aspirates for rapid diagnosis of respiratory viruses in children. J. Clin. Virol. 42, 65–69 (2008).22.R: a language and environment for statistical computing (R Foundation for Statistical Computing, Vienna, Austria, 2019).23.Yee, T. W. Vector Generalized Linear and Additive Models: with an Implementation in R (Springer, 2016).Download referencesAcknowledgementsThis work was supported by the General Research Fund of the University Grants Committee (grant no. 765811), the Health and Medical Research Fund (grant no. 13120592) and a commissioned grant of the Food and Health Bureau and the Theme-based Research Scheme (project no. T11-705/14-N) of the Research Grants Council of the Hong Kong SAR Government. We acknowledge colleagues including R. O. P. Fung, A. K. W. Li, T. W. Y. Ng, T. H. C. So, P. Wu and Y. Xie for technical support in preparing and conducting this study and enrolling participants; J. K. M. Chan, S. Y. Ho, Y. Z. Liu and A. Yu for laboratory support; S. Ferguson, W. K. Leung, J. Pantelic, J. Wei and M. Wolfson for technical support in constructing and maintaining the G-II device; V. J. Fang, L. M. Ho and T. T. K. Lui for setting up the database; and C. W. Y. Cheung, L. F. K. Cheung, P. T. Y. Ching, A. C. H. Lai, D. W. Y. Lam, S. S. Y. Lo, A. S. K. Luk and other colleagues at the Outpatient Center and Infection Control Team of Hong Kong Baptist Hospital for facilitating this study.Author informationAuthor notesThese authors jointly supervised this work: Donald K. Milton, Benjamin J. Cowling.AffiliationsWHO Collaborating Centre for Infectious Disease Epidemiology and Control, School of Public Health, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, ChinaNancy H. L. Leung, Daniel K. W. Chu, Eunice Y. C. Shiu, Benien J. P. Hau, Hui-Ling Yen, Dennis K. M. Ip, J. S. Malik Peiris, Wing-Hong Seto, Gabriel M. Leung & Benjamin J. CowlingDepartment of Microbiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, ChinaKwok-Hung ChanDepartment of Environmental Health, Harvard School of Public Health, Boston, MA, USAJames J. McDevittDepartment of Surgery, Queen Mary Hospital, Hong Kong, ChinaBenien J. P. HauDepartment of Mechanical Engineering, The University of Hong Kong, Hong Kong, ChinaYuguo LiDepartment of Pathology, Hong Kong Baptist Hospital, Hong Kong, ChinaWing-Hong SetoMaryland Institute for Applied Environmental Health, School of Public Health, University of Maryland, College Park, MD, USADonald K. MiltonAuthorsNancy H. L. LeungYou can also search for this author in Daniel K. W. ChuYou can also search for this author in Eunice Y. C. ShiuYou can also search for this author in Kwok-Hung ChanYou can also search for this author in James J. McDevittYou can also search for this author in Benien J. P. HauYou can also search for this author in Hui-Ling YenYou can also search for this author in Yuguo LiYou can also search for this author in Dennis K. M. IpYou can also search for this author in J. S. Malik PeirisYou can also search for this author in Wing-Hong SetoYou can also search for this author in Gabriel M. LeungYou can also search for this author in Donald K. MiltonYou can also search for this author in Benjamin J. CowlingYou can also search for this author in ContributionsAll authors meet the International Committee of Medical Journal Editors criteria for authorship. The study protocol was drafted by N.H.L.L. and B.J.C. Data were collected by N.H.L.L., E.Y.C.S. and B.J.P.H. Laboratory testing was performed by D.K.W.C. and K.-H.C. Statistical analyses were conducted by N.H.L.L. N.H.L.L. and B.J.C. wrote the first draft of the manuscript, and all authors provided critical review and revision of the text and approved the final version.Corresponding authorCorrespondence to\n Benjamin J. Cowling.Ethics declarations\n \n Competing interests\n B.J.C. consults for Roche and Sanofi Pasteur. The authors declare no other competing interests.\n \n Additional informationPeer review information Alison Farrell was the primary editor on this article and managed its editorial process and peer review in collaboration with the rest of the editorial team.Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.Extended dataExtended Data Fig. 1Participant enrolment, randomization of mask intervention and identification of respiratory virus infection.Extended Data Fig. 2 Weekly number of respiratory virus infections identified by RT-PCR in symptomatic individuals who had provided exhaled breath samples (respiratory droplets and aerosols) during the study period.Blue, coronavirus; red, influenza virus; yellow, rhinovirus; green, other respiratory viruses including human metapneumovirus, parainfluenza virus, respiratory syncytial virus and adenovirus; white, no respiratory virus infection identified.Extended Data Fig. 3 Respiratory virus shedding in (a) nasal swab, (b) throat swab, (c) respiratory droplets and (d) aerosols in symptomatic individuals with coronavirus NL63, coronavirus OC43, coronavirus HKU1, influenza A and influenza B virus infection.For nasal swabs and throat swabs, all infected individuals identified by RT-PCR in any collected samples were included: coronavirus NL63 (n = 8), coronavirus OC43 (n = 5), coronavirus HKU1 (n = 4), influenza A virus (n = 31) and influenza B virus (n = 14). For respiratory droplets and aerosols, only infected individuals who provided exhaled breath samples while not wearing a surgical face mask were included: coronavirus NL63 (n = 3), coronavirus OC43 (n = 3), coronavirus HKU1 (n = 4), influenza A virus (n = 19) and influenza B virus (n = 6). The box plots indicate the median with the interquartile range (lower and upper hinge) and ± 1.5 × interquartile range from the first and third quartile (lower and upper whisker). Dark blue, coronavirus NL63; light blue, coronavirus OC43; brown, coronavirus HKU1; red, influenza A virus; orange, influenza B virus.Extended Data Fig. 4 Efficacy of surgical face masks in reducing respiratory virus shedding in respiratory droplets and aerosols of symptomatic individuals with seasonal coronaviruses including (a) coronavirus NL63, (b) coronavirus OC43 and (c) coronavirus HKU1.The figure shows the virus copies per sample collected in nasal swab (red), throat swab (blue), respiratory droplets collected for 30 min while not wearing (dark green) or wearing (light green) a surgical face mask and aerosols collected for 30 min while not wearing (brown) or wearing (orange) a face mask, collected from individuals with acute respiratory symptoms who were positive for coronavirus NL63, coronavirus OC43 and coronavirus HKU1 as determined by RT-PCR in any samples. P values for mask intervention as predictor of log10 virus copies per sample in an unadjusted univariate Tobit regression model which allowed for censoring at the lower limit of detection of the RT-PCR assay are shown, with significant differences in bold. For nasal swabs and throat swabs, all infected individuals were included (coronavirus NL63, n = 8; coronavirus OC43, n = 5; coronavirus HKU1, n = 4). For respiratory droplets and aerosols, numbers of infected individuals who provided exhaled breath samples while not wearing or wearing a surgical face mask, respectively were: coronavirus NL63 (n = 3 and 5), coronavirus OC43 (n = 3 and 4), coronavirus HKU1 (n = 4 and 2). A subset of participants provided exhaled breath samples for both mask interventions (coronavirus NL63, n = 0; coronavirus OC43, n = 2; coronavirus HKU1, n = 2).Extended Data Fig. 5 Efficacy of surgical face masks in reducing respiratory virus shedding in respiratory droplets and aerosols of symptomatic individuals with seasonal influenza viruses including (a) influenza A and (b) influenza B virus.The figure shows the virus copies per sample collected in nasal swab (red), throat swab (blue), respiratory droplets collected for 30 min while not wearing (dark green) or wearing (light green) a surgical face mask and aerosols collected for 30 min while not wearing (brown) or wearing (orange) a face mask, collected from individuals with acute respiratory symptoms who were positive for influenza A and influenza B virus as determined by RT-PCR in any samples. P values for mask intervention as predictor of log10 virus copies per sample in an unadjusted univariate Tobit regression model which allowed for censoring at the lower limit of detection of the RT-PCR assay are shown, with significant differences in bold. For nasal swabs and throat swabs, all infected individuals were included (influenza A virus, n = 31; influenza B virus, n = 14). For respiratory droplets and aerosols, numbers of infected individuals who provided exhaled breath samples while not wearing or wearing a surgical face mask, respectively were: influenza A virus (n = 19 and 19), influenza B virus (n = 6 and 10). A subset of participants provided exhaled breath samples for both mask interventions (influenza A virus, n = 7; influenza B virus, n = 2).Extended Data Fig. 6 Correlation of coronavirus viral shedding between different samples (nasal swab, throat swab, respiratory droplets and aerosols) in symptomatic individuals with seasonal coronavirus infection.For nasal swabs and throat swabs, all infected individuals were included (n = 17). For respiratory droplets and aerosols, only infected individuals who provided exhaled breath samples while not wearing a surgical face mask were included (n = 10). r, the Spearman’s rank correlation coefficient.Extended Data Fig. 7 Correlation of influenza viral shedding between different samples (nasal swab, throat swab, respiratory droplets and aerosols) in symptomatic individuals with seasonal influenza infection.For nasal swabs and throat swabs, all infected individuals were included (n = 43). For respiratory droplets and aerosols, only infected individuals who provided exhaled breath samples while not wearing a surgical face mask were included (n = 23). r, the Spearman’s rank correlation coefficient.Extended Data Fig. 8 Correlation of rhinovirus viral shedding between different samples (nasal swab, throat swab, respiratory droplets and aerosols) in symptomatic individuals with rhinovirus infection.For nasal swabs and throat swabs, all infected individuals were included (n = 54). For respiratory droplets and aerosols, only infected individuals who provided exhaled breath samples while not wearing a surgical face mask were included (n = 36). r, the Spearman’s rank correlation coefficient.Extended Data Fig. 9 Respiratory virus shedding in respiratory droplets and aerosols stratified by days from symptom onset for (a) coronavirus, (b) influenza virus or (c) rhinovirus.The figures shows the virus copies per sample collected in nasal swab (red), throat swab (blue), respiratory droplets (dark green) and aerosols (brown) collected for 30 min while not wearing a surgical face mask, stratified by the number of days from symptom onset on which the respiratory droplets and aerosols were collected. For nasal swabs and throat swabs, all infected individuals were included (coronavirus, n = 17; influenza virus, n = 43; rhinovirus, n = 54). For respiratory droplets and aerosols, numbers of infected individuals who provided exhaled breath samples while not wearing or wearing a surgical face mask, respectively were: coronavirus (n = 10 and 11), influenza virus (n = 23 and 28), rhinovirus (n = 36 and 32). A subset of participants provided exhaled breath samples for both mask interventions (coronavirus, n = 4; influenza virus, n = 8; rhinovirus, n = 14). The box plots indicate the median with the interquartile range (lower and upper hinge) and ± 1.5 × interquartile range from the first and third quartile (lower and upper whisker).Supplementary informationAbout this articleCite this articleLeung, N.H.L., Chu, D.K.W., Shiu, E.Y.C. et al. Respiratory virus shedding in exhaled breath and efficacy of face masks.\n Nat Med (2020). https://doi.org/10.1038/s41591-020-0843-2Download citationReceived: 02 March 2020Accepted: 20 March 2020Published: 03 April 2020DOI: https://doi.org/10.1038/s41591-020-0843-2"],["topImageUrl",null],["error",null]]},{"__iterable":"Map","data":[["title","職安系副教授實測「乾蒸口罩」5次內過濾效力仍合格，但強調僅應急不建議長期使用 - The News Lens 關鍵評論網"],["url","https://www.thenewslens.com/article/131064"],["summary","武漢肺炎疫情導致國內口罩需求量增大，網路上也出現各種可以讓口罩重複使用的「復活偏方」，包括用電鍋乾蒸、用紫外線照、用酒精消毒等。如今，中山醫學大學職安系副教授賴全裕表示，經過檢測證實，「電鍋乾蒸」口罩，5次以內，過濾效果都還在規定合格的80%以內。但中央流行疫情指揮中心強調，以台灣疫情狀況，非慢性病患，在開放空間不用戴口罩。《中央社》報導，賴全裕表示，「電鍋乾蒸法」可使口罩復活。電鍋乾蒸法可以利用一般家用電鍋、電鍋架及內鍋，在電鍋內依序置入內鍋架、內鍋，並放入沒沾上體液、血液或鼻涕的口罩，不用加水，直接按下煮飯按鍵，乾蒸約2到3分鐘，等到煮飯鍵跳起後，再繼續保溫約5分鐘，即可拿出來繼續使用。賴全裕指出，一般經認證合格的外科口罩，過濾效力約為80%，乾蒸5次後，過濾效力減損到70%以下，這時就必須丟棄，不能再使用了。但賴全裕也提醒，這是口罩短缺時的應急措施，不建議長期使用。根據中山醫學大學特色領域研究中心介紹，賴全裕研究專長為個人防護具（包含呼吸防護具、防護衣等）進行生物防護性能及消毒、再利用。此外，《自由時報》訪問亞東紀念醫院感染管制中心主任廖俊星，廖俊星表示，紫外線消毒、水蒸口罩、酒精消毒，都沒有研究證實能有效消毒口罩，其中，水蒸、酒精消毒還可能破壞口罩結構。但曾有人拿過期口罩測驗，如果口罩沒有被破壞或使用，證實效果還在。此外，昨日，中央流行疫情指揮中心的記者會上，也有記者提問表示，有台大醫師認為口罩用越久，效果越好。指揮中心醫療整備官羅一鈞澄清，這有違一般對於口罩的學理跟常識，有待證實。指揮中心昨日也再次呼籲，開放空間不用戴口罩，但去醫院看病、出入人多擁擠地方、慢性病患要戴口罩。此外，也要以肥皂肥皂勤洗手、避免觸摸眼鼻口。 \t \t\t \t\t\t消毒方法 \t\t\t是否證實有效 \t\t\t資訊來源 \t\t \t\t \t\t\t電鍋乾蒸 \t\t\t是 \t\t\t（乾蒸口罩超過5次即需丟棄） \t\t\t中山醫學大學職安系副教授賴全裕 \t\t \t\t \t\t\t紫外線消毒 \t\t\t未證實 \t\t\t亞東紀念醫院感染管制中心主任廖俊星 \t\t \t\t \t\t\t水蒸 \t\t\t未證實 \t\t\t亞東紀念醫院感染管制中心主任廖俊星 \t\t \t\t \t\t\t酒精消毒 \t\t\t未證實 \t\t\t亞東紀念醫院感染管制中心主任廖俊星 \t\t \t\t \t\t\t過期口罩 \t\t\t是 \t\t\t亞東紀念醫院感染管制中心主任廖俊星 \t\t \t\t \t\t\t口罩用越久效果越好 \t\t\t未證實 \t\t\t中央流行疫情指揮中心醫療整備官羅一鈞 \t\t \t 延伸閱讀： \t【插畫】「武漢肺炎」來勢洶洶，口罩該買哪種？ \t口罩實名制上路：如何買？怎麼查哪還有庫存？8個QA一次看懂 \t其實你不一定要戴口罩：武漢肺炎讓全台陷「口罩之亂」，6個QA看怎麼選、如何用新聞來源： \t口罩復活術真有效？醫師破解防疫網路謠言（自由時報） \t口罩電鍋乾蒸3分鐘殺菌學者實測最多重複用5次（中央社）責任編輯：李修慧"],["topImageUrl",null],["error",null]]},{"__iterable":"Map","data":[["title","認證電鍋可以乾蒸口罩！指揮中心︰殺菌實驗達標 - 生活 - 自由時報電子報"],["url","https://news.ltn.com.tw/news/life/breakingnews/3123669"],["summary","針對口罩使用電鍋乾蒸延長使用壽命進行實驗，結果發現無論細菌過濾效率或微粒過濾效率都跟原本口罩效果相近。（指揮中心提供）\n \n\n\n 2020-04-05 16:45:51〔記者簡惠茹／台北報導〕衛福部食藥署和中山醫學大學職業安全衛生系副教授賴全裕都針對口罩使用電鍋乾蒸延長使用壽命進行實驗，實驗結果發現無論細菌過濾效率或微粒過濾效率都跟原本口罩效果相近。指揮官陳時中指出，從這禮拜四4月9日開始，口罩就可以14天買9片，這個量應該是足夠，還是非常期待大家撙節使用，食藥署也有提供增加口罩使用次數或延長使用時間的方法。 \n \n食藥署說明電鍋乾蒸方法，一，先將內鍋架及內鍋依序放入乾燥電鍋；二、將醫用口罩平放於內鍋中，蓋上外鍋蓋；三、按下電鍋加熱開關，等約3分鐘待開關自動跳起後，再靜置5分鐘，使鍋內熱空氣充分殺菌；四、打開外鍋蓋，完成殺菌程序，待電鍋冷卻後再進行下一次口罩殺菌。指揮中心研究組副組長吳秀梅說明，自己使用過的醫用外科口罩如果想要延長壽命，可以放入電鍋內鍋架乾蒸，提醒不要放水，放水會影響口罩靜電層，有人問是否可以使用酒精消毒，其實也不好，因為靜電層會受到影響。吳秀梅也提醒，如果口罩有髒汙破損不建議再度消毒使用，有呼吸道症狀、慢性疾病、出入醫院以一次使用為原則，低風險場所就會建議可以使用布口罩，有不織布層的布口罩，可以重複清洗效果也好，也可以使用電鍋乾蒸重複消毒4到5次。吳秀梅說明外科口罩測試部分，除了有參考學研單位建議，食藥署也做過研究，她表示，經過3到5次消毒後，對於細菌過濾效率（BFE）可以維持99%以上。中山醫學大學副教授賴全裕實驗室做過電鍋乾蒸口罩實驗，他指出，粒子過濾效果（PFE）檢測中，儀器會產生2奈米到2微米的微粒，其中99%是10奈米到770奈米的微粒，然後來看口罩可以過濾多少，總過濾效果如果超過95%，就是N95口罩，80%以上是外科手術口罩，電鍋乾蒸實驗結果發現，第一次乾蒸跟原本外科口罩過濾效果相比還能維持99%，只是纖維帶電還是有些微影響，但是消毒越多次，就會越下降，建議不要超過5次。至於病毒過濾效果，賴全裕說，雖然外科手術口罩沒有檢測病毒過濾效果，主要是防護衣才會測試，但是武漢肺炎病毒的大小大概是100奈米，所以檢測PFE就有將病毒尺寸涵蓋在內。賴全裕也解答民眾各種電鍋蒸口罩的疑慮。許多民眾詢問可以使用家裡的電鍋就好嗎？還是要買新的電鍋？賴全裕指出，由於內鍋壁165度左右，該殺死的細菌應該都殺死了，乾蒸氣溫大概110度左右，如果有疑慮可以洗鍋子。口罩氣味太重怎麼辦？可以加精油或酒精嗎？賴全裕表示，有化學物質的都會破壞口罩不織布纖維帶電，不建議這樣做，因為味道來源是口水和細菌繁殖的關係，殺死後理論上味道就會不見。用烤箱、氣炸鍋或微波爐可以嗎？賴全裕指出，這些家電如果可以控制溫度達110度，可以模擬電鍋溫度的話就可以嘗試，但是還是要實測才知道，微波爐可能會出現口罩燃燒要小心。使用負離子吹風機可以嗎？賴全裕表示，目前還沒有實驗檢測，暫時不建議。殺菌燈小型烘碗機可以嗎？賴全裕指出，如果紫外燈波長有220奈米到280奈米，可是口罩裡面有3層，紫外燈只能對上下兩面照射，不確定中間層的殺菌效果，使用的時候還是建議要注意，未來有機會會再做實驗。用煮的可以嗎？賴全裕說，還沒有實驗可以確認。暫時拿下來的口罩如何收納？賴全裕指出，建議把口罩放在通風乾燥處，拿下來的時候，不要使用完全密封的塑膠袋，會造成細菌容易繁殖，可以用塑膠夾或紙袋，避免手觸摸到口罩。相關新聞請見︰武漢肺炎懶人包》傳染途徑、預防措施看這裡「武漢肺炎專區」請點此，更多相關訊息，帶您第一手掌握。\n\n \n\n\n\n \n\n \n 不用抽不用搶現在用APP看新聞保證天天中獎　\n 點我下載APP　\n 按我看活動辦法\n\n \n \n\n\n\n 相關新聞\n \n \n\n\n\n\n\n\n\n\n \n \n\n\n\n\n\n\n\n\n\n\n LTN集點大放送\n \n \n \n \n \n \n\n\n\n\n\n網友回應"],["topImageUrl",null],["error",null]]},{"__iterable":"Map","data":[["title","Virological assessment of hospitalized patients with COVID-2019"],["url","https://www.nature.com/articles/s41586-020-2196-x"],["summary","AbstractCoronavirus disease 2019 (COVID-19) is an acute respiratory tract infection that emerged in late 20191,2. Initial outbreaks in China involved 13.8% cases with severe, and 6.1% with critical courses3. This severe presentation corresponds to the usage of a virus receptor that is expressed predominantly in the lung2,4. By causing an early onset of severe symptoms, this same receptor tropism is thought to have determined pathogenicity, but also aided the control, of severe acute respiratory syndrome (SARS) in 20035. However, there are reports of COVID-19 cases with mild upper respiratory tract symptoms, suggesting the potential for pre- or oligosymptomatic transmission6–8. There is an urgent need for information on body site-specific virus replication, immunity, and infectivity. Here we provide a detailed virological analysis of nine cases, providing proof of active virus replication in upper respiratory tract tissues. Pharyngeal virus shedding was very high during the first week of symptoms (peak at 7.11 × 108 RNA copies per throat swab, day 4). Infectious virus was readily isolated from throat- and lung-derived samples, but not from stool samples, in spite of high virus RNA concentration. Blood and urine never yielded virus. Active replication in the throat was confirmed by viral replicative RNA intermediates in throat samples. Sequence-distinct virus populations were consistently detected in throat and lung samples from the same patient, proving independent replication. Shedding of viral RNA from sputum outlasted the end of symptoms. Seroconversion occurred after 7 days in 50% of patients (14 days in all), but was not followed by a rapid decline in viral load. COVID-19 can present as a mild upper respiratory tract illness. Active virus replication in the upper respiratory tract puts the prospects of COVID-19 containment in perspective.\n \n \n \n\n \n \n \n \n\n Author informationAuthor notesThese authors contributed equally: Roman Wölfel, Victor M. Corman, Wolfgang GuggemosThese authors jointly supervised this work: Christian Drosten, Clemens WendtnerAffiliationsBundeswehr Institute of Microbiology, Munich, GermanyRoman Wölfel, Sabine Zange, Patrick Vollmar, Rosina Ehmann & Katrin ZwirglmaierCharité Universitätsmedizin Berlin, Berlin, GermanyVictor M. Corman, Marcel A. Müller, Daniela Niemeyer, Terry C. Jones, Tobias Bleicker, Sebastian Brünink, Julia Schneider & Christian DrostenKlinikum München-Schwabing, Munich, GermanyWolfgang Guggemos, Michael Seilmaier & Clemens WendtnerCenter for Pathogen Evolution, Department of Zoology, University of Cambridge, Cambridge, UKTerry C. JonesUniversity Hospital LMU Munich, Munich, GermanyCamilla Rothe & Michael HoelscherAuthorsRoman WölfelYou can also search for this author in Victor M. CormanYou can also search for this author in Wolfgang GuggemosYou can also search for this author in Michael SeilmaierYou can also search for this author in Sabine ZangeYou can also search for this author in Marcel A. MüllerYou can also search for this author in Daniela NiemeyerYou can also search for this author in Terry C. JonesYou can also search for this author in Patrick VollmarYou can also search for this author in Camilla RotheYou can also search for this author in Michael HoelscherYou can also search for this author in Tobias BleickerYou can also search for this author in Sebastian BrüninkYou can also search for this author in Julia SchneiderYou can also search for this author in Rosina EhmannYou can also search for this author in Katrin ZwirglmaierYou can also search for this author in Christian DrostenYou can also search for this author in Clemens WendtnerYou can also search for this author in Corresponding authorsCorrespondence to\n Christian Drosten or Clemens Wendtner.Supplementary information\n \n \n Supplementary InformationThis file contains Supplementary Methods, which include a description of RNA extraction and RT-PCR methods, a description of cell culture and antibody detection methods and Supplementary References.About this articleCite this articleWölfel, R., Corman, V.M., Guggemos, W. et al. Virological assessment of hospitalized patients with COVID-2019.\n Nature (2020). https://doi.org/10.1038/s41586-020-2196-xDownload citationReceived: 01 March 2020Accepted: 24 March 2020Published: 01 April 2020DOI: https://doi.org/10.1038/s41586-020-2196-x\n\n \n\n \n CommentsBy submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate."],["topImageUrl",null],["error",null]]}]}],["text","#口罩可以乾蒸嗎\n#為何2月不行現在可以\n#水鏡來告訴你\n\n今天疾管中心宣傳口罩可以乾蒸，再重複使用\n馬上有人找出2月衛福部食藥署建議不要乾蒸的資料\n還有人把這照片放在爆料公社，很明顯要帶風向\n\n有人酸因為唐鳳說可以乾蒸，所以政策就轉彎\n\n不管別人怎麼說，包括這篇都是\n請做個獨立思考的人\n\n#防疫本來就是與時俱進\n大家應該還記得，2月的時候疾管中心跟大家說不用戴口罩\n總統、行政院長、陳時中部長，也帶頭不戴口罩\n當時還有「我OK，你先戴」的風氣\n\n當時背景是沒有社區感染，戴不戴口罩風險都低\n當時到現在沒戴口罩的人，都活得好好的也沒被感染。\n\n這兩天陳時中部長已經戴起口罩，為什麼呢？\n簡單說，我們有零星的社區感染，加上這幾天連假的出遊\n我們要在未來的14天防範疫情社區化\n\nNature Medicine刊出香港學者研究，外科口罩可以顯著防止新冠病毒從感染者傳播出去。https://www.nature.com/articles/s41591-020-0843-2?fbclid=IwAR3EzP6Xm2bXH4bDHx9i9oOYxqHIraWbuRbZcGzM6fmgQz7iIl7ufHrkN-A\n\n這是因為疫情的不同，資訊的改變\n政府反應以及需要民間配合的方式不同\n\n#那到底口罩可不可以乾蒸\n\n你可能會問，當時食藥署為何覺得不行？\n當時食藥署的說法是\"可能\"會破壞結構，所以不建議。\n\n但後續的研究出來，真的可以透過乾蒸讓口罩重複使用\n\n根據中山醫學大學職安系副教授賴全裕表示，經過檢測證實，「電鍋乾蒸」口罩，5次以內，過濾效果都還在規定合格的80%以內。\nhttps://www.thenewslens.com/article/131064\n\n實驗結果發現無論細菌過濾效率或微粒過濾效率都跟原本口罩效果相近。\nhttps://news.ltn.com.tw/news/life/breakingnews/3123669\n\n再說一次，這是人類第一次遇到這個病毒，很多事情本來就不了解。\n\n比如Nature這篇研究，就刷新大家原本的認知\n\n原來剛被感染的前5天，傳染力最大，到住院第9天傳染力就低了，不一定要在負壓房。\n\n這對本來以為症狀越嚴重傳播力越大、負壓房要住好住滿的觀念是很大的釐清\nhttps://www.nature.com/articles/s41586-020-2196-x\n\n好，回到乾蒸口罩這件事\n原先政府有疑惑，後來研究證實，口罩乾蒸可以重複使用\n政府提供資訊，不OK嗎？\n\n#未來14天大家照顧好自己\n\n現在已經有人在傳\n「為了要向國外捐口罩，所以要求民眾乾蒸」\n\n請問我們有為了捐口罩，一周從買三個變買兩個嗎？\n\n提供乾蒸，只是資訊更新後提供方法\n可能有些人不覺得一個口罩五元貴\n\n但對節省、重視環保的或有經濟壓力的人，這不失為權宜之計\n至於要酸的、要戰的、要攻擊的，\n就別了，台灣需要大家的團結\n\n就像這幾天的連假，\n大家玩都玩了，該得的也得了，不該得的也不會得\n\n本院只有一個請求\n\n#連假有出遊的朋友接下來14天外出請戴口罩\n\n接下來14天，大家把自己照顧好\n勤洗手、出門戴口罩、保持人際距離、不摸臉、少聚集\n\n#祝大家平安健康\n\n#我們二月也不知道肥胖容易得重症啊\n\n#水鏡共筆"],["id","1kyvy3pelrgow"],["createdAt","2020-04-06T08:20:58.169Z"]]}],["reply",{"__iterable":"Map","data":[["id","yQweUHEBrhVJn3LNh3pF"],["user",{"__iterable":"Map","data":[["id","_1YnAG8BV0DITCqGm7go"],["name","uienwt"],["avatarUrl",null],["level",18]]}],["type","OPINIONATED"],["text","為何2月不行現在可以，陳時中說，他一直很期待可以研發讓口罩重複使用的方法，但食藥署站在專業角度，堅持要經過驗證後才正式公告，他也認為這是對的。"],["reference","https://newtalk.tw/news/view/2020-04-06/386656\n電鍋蒸口罩為何延遲公布？"],["hyperlinks",{"__iterable":"List","data":[{"__iterable":"Map","data":[["title","武漢肺炎》電鍋蒸口罩為何延遲公布？陳時中說明原因 | 社會 | 新頭殼 Newtalk"],["url","https://newtalk.tw/news/view/2020-04-06/386656"],["summary","中央流行疫情指揮中心指揮官陳時中（左起）與食藥署署長吳秀梅，5日示範以電鍋乾蒸口罩。圖：翻攝自疾管署直播影片 \n \n \n \n \n \n \n 2月的時候，指揮中心對電鍋蒸口罩重複消毒一事並未正式證實其效用，直到4月5日，陳時中與食藥署署長吳秀梅親自在記者會上示範用電鍋乾蒸消毒口罩。陳時中說，他一直很期待可以研發讓口罩重複使用的方法，但食藥署站在專業角度，堅持要經過驗證後才正式公告，他也認為這是對的。\n昨天記者會上，陳時中與吳秀梅現場使用電鍋消毒口罩，將口罩放進電鍋乾蒸3分鐘，開關跳起後等待5分鐘，消毒程序就大功告成。吳秀梅表示，電鍋乾蒸法消毒的口罩可維持99%的細菌過濾效率，讓口罩重複使用。\n陳時中表示，他一直很期待研發出讓口罩重複使用的有效方法，當他知道電鍋蒸口罩有相關研究報告佐證時也想推廣這種方式，但食藥署站在專業角度，認為沒實證不夠謹慎。食藥署花時間對口罩進行檢測實驗，驗證完之後才公布這是有效方法且容易執行。\n陳時中也同意食藥署的看法，認為他們用更嚴謹的態度處理是對的，因此昨天才正式公布。 \n \n \n \n 他一直很期待可以研發讓口罩重複使用的方法但食藥署站在專業角度，堅持要經過驗證後才正式公告直到4月5日，陳時中與食藥署署長吳秀梅親自在記者會上示範用電鍋乾蒸消毒口罩。"],["topImageUrl",null],["error",null]]}]}],["createdAt","2020-04-06T15:31:50.222Z"]]}]]},{"__iterable":"Map","data":[["article",{"__iterable":"Map","data":[["replyCount",1],["references",{"__iterable":"List","data":[{"__iterable":"Map","data":[["type","LINE"]]}]}],["replyRequestCount",1],["user",{"__iterable":"Map","data":[["id","j4S8C_AO7U9UgO3jfO7w-_TCIO4Xtij_63hrFiTOAIF-wiO0o"],["name","忠誠的✖長濱✖法蘭克林"],["avatarUrl",null],["level",0]]}],["hyperlinks",{"__iterable":"List","data":[{"__iterable":"Map","data":[["title","Universal and reusable virus deactivation system for respiratory protection"],["url","https://www.nature.com/articles/srep39956#Sec8"],["summary","AbstractAerosolized pathogens are a leading cause of respiratory infection and transmission. Currently used protective measures pose potential risk of primary/secondary infection and transmission. Here, we report the development of a universal, reusable virus deactivation system by functionalization of the main fibrous filtration unit of surgical mask with sodium chloride salt. The salt coating on the fiber surface dissolves upon exposure to virus aerosols and recrystallizes during drying, destroying the pathogens. When tested with tightly sealed sides, salt-coated filters showed remarkably higher filtration efficiency than conventional mask filtration layer, and 100% survival rate was observed in mice infected with virus penetrated through salt-coated filters. Viruses captured on salt-coated filters exhibited rapid infectivity loss compared to gradual decrease on bare filters. Salt-coated filters proved highly effective in deactivating influenza viruses regardless of subtypes and following storage in harsh environmental conditions. Our results can be applied in obtaining a broad-spectrum, airborne pathogen prevention device in preparation for epidemic and pandemic of respiratory diseases.\n \n \n \n \n \n \n\n \n IntroductionAerosols take a prominent role in airborne transmission of respiratory diseases. Droplets with aerodynamic size (da) < 10 μm and 10 < da < 100 μm are known to infect the alveolar regions and upper respiratory tract, respectively1,2. Notably, aerosols can also be a route of infection in diseases that, contrary to for instance influenza, do not specifically target the respiratory tract, as it could be the case of Ebola virus3. While vaccination can greatly reduce morbidity and mortality, during a pandemic or epidemic new vaccines matching the specific strain would be available, at the earliest, six months after the initial outbreak. Additionally, following development of an effective viral vaccine, several potential problems would remain, such as limited supply due to insufficient production capacity and time-consuming manufacturing processes. As a result, individuals close to the point of an outbreak would be in imminent danger of exposure to infectious diseases during the non-vaccine period. In the absence of vaccination, respirators and masks can be worn to prevent transmission of airborne pathogenic aerosols and control diseases, such as influenza4.The main alternative, the N95 respirator, requires training prior to use, must be expertly fitted to address the risk of faceseal leakage at the face-mask interface, and must be disposed of as biohazard5. Due to these factors, the use of N95 respirators on a large scale is impractical and expensive during an epidemic or pandemic. Past experiences of severe acute respiratory syndrome (SARS), H1N1 swine flu in 2009, and Middle East respiratory syndrome (MERS) indicate that surgical masks have been most widely adopted by the public as personal protective measure, despite controversy on their effectiveness6,7,8,9. Currently, among other factors, filtration in respirators and masks depends on filter characteristics, including fiber diameter, packing density, charge of fibers and filter thickness, as well as particle properties, such as diameter, density and velocity10,11,12,13,14. However, in the lack of a system to deactivate the collected pathogens, safety concerns naturally arise about secondary infection and contamination from virus-laden filter media during utilization and disposal. Furthermore, since re-sterilization is not possible without causing damage, respirators and masks are recommended for single use only9,15,16. Scientific efforts have been focused on treatment of filters with materials possessing well-known antimicrobial properties, such as iodine, chlorine and metals17,18,19,20,21,22,23,24,25, although with limited effectiveness against virus aerosols26,27,28. Therefore, a key challenge is the development of an easy-to-use, universal virus negation system, which is reusable without reprocessing and capable of deactivating pathogens, thereby reducing potential risk of secondary infection and transmission.Here, we report a simple but efficient virus inactivation system exploiting the naturally occurring salt recrystallization. Our strategy is to modify the surface of the fibrous filtration layer within masks with a continuous salt film for virus deactivation via two successive processes: i) salt is locally dissolved by the viral aerosols and ii) supersaturation is followed by evaporation-induced salt recrystallization. Consequently, viruses are exposed to increasingly higher concentrations of saline solution during drying and physically damaged by recrystallization.ResultsPreparation and characterization of salt-functionalized filtersTo demonstrate the concept of virus deactivation system based on salt recrystallization, the middle layer of three-ply surgical mask, polypropylene (PP) microfiber filter, was coated with NaCl salt as an active virus negation unit (see Supplementary Fig. S1 for bare PP filter). The coating formulations contained surfactant to enhance wetting of saline solution on the surface of hydrophobic PP fibers. Bare PP filters (abbreviated as Filterbare) were pre-wet to contain about 600 μL of coating solution (abbreviated as Filterwet). The amount of NaCl salt (Wsalt in mg/cm2) coated on the filter per unit area, considering that the filters thickness is constant, was easily controlled by changing the coating solution volume (Vsalt in μL) during drying of pre-wet filter (radius: 3 cm, Wsalt = 3.011 + 0.013 × Vsat, n = 7) (Fig. S2). Scanning electron microscopy (SEM) and energy dispersive X-ray (EDX) mapping analysis showed the formation of homogeneous NaCl coating during drying, as also confirmed by X-ray diffraction (XRD) (Fig. 1a,b and Supplementary Fig. S3). Both the formation of NaCl coating on PP fibers and presence of surfactant in the coating formulation appeared to alter the filter surface properties from hydrophobic (bare filter; contact angle, θc = 133.0 ± 4.7°) to completely hydrophilic (salt-coated filter; θc ~ 0°, n = 10) (Fig. 1c and Supplementary Fig. S4). Hydrophilic nature of salt coating can greatly improve adhesion of viral aerosols to PP fibers compared to Filterbare, as seen in Raman microscope images (Fig. 1d and Supplementary Fig. S5).Figure 1: Mask with salt-coated filter for prevention and deactivation of airborne pathogens.(a) SEM image of Filterwet+600μL (top left) and EDX mapping images of Na (red), Cl (green), and NaCl (combination of Na and Cl mapping images), showing the formation of NaCl coating, as also confirmed by XRD spectra (b) of Filterbare, Filterwet, Filterwet+100μL, Filterwet+300μL, Filterwet+600μL, Filterwet+900μL and Filterwet+1200μL (labelled as Bare, wet, wet+100 μL, wet+300 μL, wet+600 μL, wet+900 μL and wet+1200 μL, respectively; miller indices corresponding to NaCl crystal are shown at the top of XRD spectra for each position). (c) Optical microscope images for contact angle measurements using 3 μL DI water droplets on (i) Filterbare and (ii) Filterwet+600μL (n = 10). (d) Microscope images of aerosol on (i) Filterbare and (ii) Filterwet+600μL (n = 10).Filtration efficiency against viral aerosols and protective efficacy in vivoFiltration efficiency of salt-coated filters was tested against aerosols with volumetric mean diameter (VMD) of 2.5–4 μm containing H1N1 pandemic influenza virus (A/California/04/2009, abbreviated as CA/09) at different pressure conditions (see Fig. 2a for transmission electron microscope (TEM) image of H1N1 virus). Interestingly, as shown in Fig. 2b, Filterbare did not exhibit any significant level of resistance against penetration of virus under our experimental conditions (i.e., 0% filtration efficiency). Conversely, salt-coated filters showed substantially increasing filtration efficiency with pressure and amount of coated salt. In particular, in the case of Filterwet+600μL, filtration efficiency varied from 43 to 70%, with increasing pressure from 3 to 17 kPa, and Filterwet+1200μL exhibited persistent, high-level efficiency (~85%) (one-way ANOVA, P = 0.85).Figure 2: Filtration efficiency of salt-coated filters.(a) TEM image of CA/09 H1N1 influenza virus. (b) Pressure-dependent filtration efficiency (n = 8–10, mean ± standard deviation (SD)). (c–f) Effects of filtration efficiency on protective efficacy in vivo. Body weight change of mice after infection with the dosages of penetrated virus (n = 12, mean ± SD) (c), survival rates (mean; 100% means that all mice in the group survived as penetrated dosages were lower than lethal dose) (d), lung virus titers (n = 4, mean ± SD) (e), and lung inflammatory cytokine (interferon-γ (IFN-γ)) assay (n = 11, mean ± SD) (f). Legends: filters are labelled as in Fig. 1b.To probe the effects of filtration efficiency on protective efficacy, in vivo experiments were performed using mice intranasally (IN) infected with penetrated dosages of H1N1 virus under breathing pressure (~10 kPa)29. As shown in Fig. 2c, similarly to negative control groups (mice infected with lethal dose of virus stock and aerosolized virus), mice exposed to a dose penetrated through the bare filter showed rapid body weight loss, followed by death within 10 days after infection, in good agreement with the observed 0% filtration efficiency (Fig. 2b). In contrast, mice groups exposed to virus derived from salt-coated filters resulted in 100% survival rate (Fig. 2d). Furthermore, lungs of mice from negative control groups exhibited severe lung infection 4 days after challenge (Fig. 2e). Conversely, mice groups exposed to virus derived from salt-coated filters showed significantly lower levels of lung viral titers (t-test, P < 0.005). This is consistent with lower levels of inflammatory cytokines, interferon-γ (IFN-γ), from salt-coated filter groups compared to negative control and bare filter groups (t-test, P < 0.001) (Fig. 2f).Deactivation of virus on salt-functionalized filtersInfluenza virus stability tests were performed to investigate the effects of salt coating. The same amount of recovered viruses from the PP fibers was used, and, in the case of bare filters, viral aerosols exposure was conducted in the absence of pressure due to 100% penetration of viral aerosols. Unlike bare filters (Fig. S6a(i)), formation of micron-sized NaCl phase represents a typical feature of salt-coated filters due to recrystallization of NaCl salt, following local dissolution upon aerosols exposure (SEM images in Fig. S6a, ii to iv, and EDX mapping in Fig. S6b). In contrast to 8% HA activity loss of virus adsorbed onto Filterbare, salt-coated filters exhibited almost complete HA activity loss within 5 min of incubation (Fig. 3a). Such dramatic virus destabilization on salt-coated filters is further supported by negligible levels of viral titers compared to Filterbare with incubation time (t-test, P < 0.001) (Fig. 3b). It is also noted that virus titers exhibited significant decrease with increase of incubation time and amount of coated salt (ANOVA general linear model, P < 0.001). TEM analysis showed that influenza virus on Filterbare experiences morphological change into non-spherical shape during aerosol drying (Fig. 3c(i)). Notably, influenza virus was severely damaged on salt-coated filters even at 5 min of incubation (Fig. 3c(ii)). From microscopic analysis, aerosol drying time was about 3 min, indicating that destruction of virus observed at 5 min is associated with drying-induced salt crystallization. Physical damage of virus due to crystallization was similarly reported as a major destabilizing factor of inactivated influenza virus30,31. Lower levels of native fluorescence and nile red fluorescence from virus recovered from salt-coated filters accounted for more severe conformational change of antigenic proteins and destabilization of viral envelope, respectively, consistent with TEM analysis (t-test, P < 0.001) (Fig. 3d). In parallel, we investigated the separate effect of salt concentration increase on virus stability during the aerosol drying process, irrespective of crystal growth. As displayed in Supplementary Fig. S7, the materials collected in suspension from Filterwet+600μL induced visible morphological transformation of the virus (Supplementary Fig. S7b) compared to suspension of Filterbare (Supplementary Fig. S7a). This can be attributed to the high salt/surfactant concentration and osmotic pressure, which have been well-known to destabilize proteins and viruses31,32,33. Therefore, the marked virus destabilization on salt-coated PP fibers can be explained by the combined effects of salt concentration increase during drying and evaporation-induced salt crystallization.Figure 3: Inactivation of virus adsorbed on salt-coated filters.(a,b) HA activity (a) and virus titer (b) displaying the effects of incubation time on the remaining activity of virus (n = 4–8, mean ± SD). (c) TEM images of viruses reconstituted, after incubation for 5 and 60 min, from (i) Filterbare and (ii) Filterwet+600μL. (d) Native fluorescence/nile red fluorescence of viruses incubated for 60 min (n = 12, mean ± SD). (e,f) Body weight change of mice after infection with virus recovered from filters after incubation for 60 min (n = 12, mean ± SD) (e), and lung virus titers (n = 6, mean ± SD) (f). Asterisk (*): below detection limit. Legends: filters are labelled as in Fig. 1b.To verify in vitro virus stability on the filters, an in vivo study was performed by infecting mice with virus incubated for 60 min on PP filters. As shown in Fig. 3e, Filterbare group exhibited 5% body weight loss at day 9 post-infection, reaching a body weight lower than that of salt-coated filter groups by 10–15%. Thus, significantly higher lung virus titers in the negative control group were observed in contrast to no detectable titers in the salt-coated filter groups (Fig. 3f).Strain-nonspecific virus deactivation and effects of storage under harsh environmental conditions on salt coating stabilityBroad-spectrum protection of salt-coated filters against multiple subtypes of viral aerosols was evaluated by investigating both lethal infectivity by penetrated virus in vivo and infectivity by virus collected on filters during filtration in vitro using A/Puerto Rico/08/1934 (PR/34 H1N1) and A/Vietnam/1203/2004 (VN/04 H5N1). Similarly to CA/09 H1N1, 100% of mice survived viral infection (PR/34 and VN/04), with no evidence of weight loss, due to higher filtration efficiency of salt-coated filter than that of bare filter (Fig. 4a). This is supported by no significant level of viral titer in the lung. In addition, as shown in Fig. 4b, salt-coated PP filters destroyed adsorbed influenza viruses irrespective of both subtypes and amount of coated salts.Figure 4: Strain- and environment-dependent performance of salt-coated filters.(a) Body weight change of mice infected with penetrated PR/34 H1N1 and VN/04 H5N1 viruses through Filterwet+600μL (n = 12, mean ± SD). (b) Virus titers of recovered viruses from bare and salt-coated filters (n = 4, mean ± SD; data for Filterwet, Filterwet+600μL and Filterwet+1200μL are overlapped). (c,d) Body weight change (c) and survival rate (d) of mice infected with dosage of penetrated virus through Filterwet+600μL before and after exposure to harsh environmental conditions (37 °C and 70% RH) for 1 day (filled square and open square overlap in (d)). (e) EDX mapping image of NaCl-coated Filterwet+600μL after incubation for 15 days at 37 °C and 70% RH (combination of Na (red) and Cl (green) mapping images). (f) XRD spectra of Filterwet+600μL before and after incubation at 37 °C 70% for 1 day and 15 days. Legends: filters are labelled as in Fig. 1b.The stability of salt coating on PP fibers was tested under harsh environmental conditions. Incubation at 37 °C and 70% relative humidity (RH) for 1 day did not cause any significant difference in filtration efficiency (t-test, P = 0.718) (Supplementary Fig. S8). As a result, all mice infected with dosage of penetrated virus through the filter stored at high temperature and RH displayed 100% survival with 7% of body weight loss (Fig. 4c,d). Even after 15 days of incubation, salts remained to coat PP fibers (Fig. 4e, and Supplementary Fig. S9a,b), despite change in grain orientation due to recrystallization (Fig. 4f, and Supplementary Fig. S10a,b).DiscussionDevelopment of a universally applicable, low-cost, and efficient mechanism for virus negation is regarded as a major challenge in public health against general airborne biological threats. This led us to propose a new concept of personal/public preventive and control measures using salt-recrystallization against pathogenic aerosols based on two hypotheses. The salt-coating can enhance adsorption of virus on the filter fibers and inactivate virus by the increase of osmotic pressure followed by the crystallization of salts. As shown in Fig. 2b, salt-coated filters exhibited significantly higher levels of filtration efficiency than bare filters. Notably, the bacterial filtration efficiency (BFE) reported by the mask manufacturer is 99%. The different value of filtration efficiency for bare filters obtained under our experimental conditions may be partially due to the use of aerosols with different biological origins. The FDA-recognized ASTM F2101 – 14 standard for evaluation of BFE exposes surgical masks to Staphylococcus aureus aerosols, by employing S. aureus ATCC 653834, which has an average diameter of about 1 μm. In this study, filtration efficiency was calculated following exposure of bare and salt-coated filters to influenza virus, which exhibits a smaller diameter than that of S. aureus by one order of magnitude. Additionally, whereas during BFE evaluation all three layers of surgical masks are used, in this work filtration efficiency refers to mask filters (middle layer). It is worth noting that the conditions for BFE standard evaluation (such as flow rate and time of application of flow) do not coincide with the experimental procedure we used for measurement of the filtration efficiency, which may further contribute to the different result. The enhanced filtration efficiency of salt-coated filters against influenza virus aerosols as compared to bare filters can be explained by the observed wetting of aerosols, favoring greater adhesion to salt-coated filters. Furthermore, the significant improvement in filtration efficiency resulted in complete protection of mice against lethal influenza aerosols, which demonstrates the high level of protection provided by salt-coated filters, outperforming currently used bare filters.Rapid loss of HA activity and viral infectivity on salt-coated filters can be explained by physical destruction of virus during recrystallization of coated salts. When the salt-coated filter is exposed to virus aerosols, salt crystals below the aerosol droplet dissolve to increase osmotic pressure to virus. Due to evaporation, the salt concentration of the droplet significantly increases and reaches the solubility limit, leading to recrystallization of salt. As a consequence, virus particles are exposed to increasing osmotic pressure during the drying process and are physically damaged by crystallization. As shown in Fig. 3e,f, the superior advantage of physically destroying the virus adsorbed to the salt-coated PP filters through natural salt crystallization process was further confirmed in vivo. According to previous reports, hyperosmotic stress (>541 mOsm) and crystallization induce membrane perturbation with irreversible deformation of the viral envelope and structural virus damage, respectively, resulting in infectivity loss of virus30,31. Therefore, our data support that the extensive level of infectivity loss associated with a salt recrystallization process caused by physical contact between virus aerosols and salt coating can be used in developing virus negation systems that are reusable without reprocessing.Similarly to CA/09 H1N1 aerosols, increased protection in vivo due to higher filtration efficiency of salt-coated filters compared to bare filters and deactivation of virus on salt-coated filters were observed following exposure to PR/34 H1N1 and VN/04 H5N1 (Fig. 4a,b). This suggests that salt-coated filters prevent virus penetration and destroy virus attached to the filter in a non-specific way. Furthermore, the performance of salt-coated filters was not degraded by storage at 37 °C and 70% RH, demonstrating that salt recrystallization-based filters can ensure protection even under harsh environmental conditions. Notably, for demonstration of the concept of salt-recrystallization based virus deactivation system, NaCl salt was used, which has a critical RH of 75% at 30 °C35. However, salts with higher critical RH can be easily used, such as ammonium sulfate, potassium chloride and potassium sulfate, which have critical RH of 80%, 84% and 96.3% at 30 °C, respectively35. This suggests that salt-coated filters may be developed for specific environmental conditions.In conclusion, we demonstrated that the developed salt-recrystallization based filtration system provides high filtration efficiency and successfully deactivates multiple subtypes of adsorbed viruses. Moreover, we have shown that stability of the salt coating is not compromised by high temperature and humidity, which suggests safe use and long-term storage/reuse at such environmental conditions. Although our tests are based on exposure to different types of influenza virus, the significance of these results for personal and public protective measures may be generally extended to enveloped respiratory viruses where infection and transmission can occur by aerosol. Our salt-coated filter unit can promise the development of long-term stable, versatile airborne pathogen negation system, without safety concerns. In fact, the destruction mechanism of viruses solely depends on the simple, yet robust naturally occurring salt recrystallization process, combining the destabilizing effects of salt crystal growth and concentration increase during drying of aerosols. This idea can be easily applied to a wide range of existing technologies to obtain low-cost, universal personal and public means of protection against airborne pathogens, such as masks and air filters in hospitals. Therefore, we believe that salt-recrystallization based virus deactivation system can contribute to global health by providing a more reliable means of preventing transmission and infection of pandemic or epidemic diseases and bioterrorism.MethodsBare and salt-coated filter samples preparationThe commercial surgical masks had a three-ply structure. The middle layer is the filter media, whereas the inner and outer layers provide support and protect the filter against wear and tear. The metal nose clips and elastic ear loops were removed and circular samples (radius: 3 cm) were cut from the masks. The PP filters (middle layer) were isolated by removing the inner and outer protective layers (bare filters, Filterbare). The coating solution was prepared by dissolving sodium chloride (NaCl; Sigma Aldrich, St. Louis, MO) in filtered DI water (0.22 μm pore size; Corning, Tewksbury, MA) under stirring at 400 rpm and 90 °C, followed by the addition of Tween 20 (Fisher Scientific) to a final concentration of 29.03 w/v% of NaCl and 1 v/v% of Tween 20. To obtain the salt-coated filters, the mask bare PP filters were pre-wet to contain approximately 600 μL of coating solution by incubating overnight at room temperature. Any remaining dry areas were removed by applying gentle strokes with tweezers to the filters while immersed in the coating solution. Subsequently, the filters were deposited in the desired volume of coating solution (0, 100, 300, 600, 900 and 1200 μL, of which corresponding membranes are abbreviated as Filterwet, Filterwet+100μL, Filterwet+300μL, Filterwet+600μL, Filterwet+900μL, and Filterwet+1200μL, respectively) on petri dishes (60 × 15 mm; Fisher Scientific) to control the amount of NaCl per unit area and dried in an oven (Isotemp Incubator, Fisher Scientific) at 37 °C for 1 day.Influenza virus preparationInfluenza viruses A/California/04/2009 (CA/09, H1N1), A/Puerto Rico/8/34 (PR/34, H1N1) and A/Vietnam/1203/2004 (VN/04, H5N1) were grown in 10-day old embryonated hen eggs, in which H5N1 virus was derived by reverse genetics from HPAI A/Vietnam/1203/200436. Influenza viruses were purified from allantoic fluid using discontinuous sucrose gradient (15%, 30% and 60%) layers following the previously reported procedure37.Aerosols exposure to filtersFor experiments involving aerosols exposure, an aerosol chamber (L × W × H = 145 × 145 × 150 mm; Emka Inc., Middletown, PA) was used (Fig. S11). It has a connection to the vacuum line and a circular aperture in the top wall (diameter: 22 mm) to exactly accommodate the cylindrical part (diameter: 20 mm, height; 20 mm) of the nebulizer unit that is below the aerosol generator (Aeroneb Lab Nebulizer System; Aerogen, Galway, Ireland). Bleach was used as trap between the chamber and the vacuum pump (Welch 2522C-10, 22 L/min; Niles, IL). The filters were placed on top of the chamber aperture and the nebulizer unit was inserted, ensuring the tight seal of the filters against the side of the aperture. 5 μL of virus stock were added to the nebulizer unit, aerosols (VMD 2.5–4 μm from manufacturer specifications) were generated for 30 sec and subsequently the desired vacuum level (3, 10 or 17 kPa) was applied, by manual control, three times in 1 sec cycles. Notably, in the case of bare filters, pressure was only applied for filtration efficiency tests.For all assays and analysis, suspensions of the filters were prepared as follows, unless otherwise indicated. To reconstitute virus adsorbed onto filters, virus-laden filters were immersed in 400 μL of sterilized DI water for about 5 min, and then removed after vortexing from the suspension. The virus suspension was centrifuged at 19,800 g and 4 °C for 10 min (Centrifuge 5810 R, Eppendorf, Hauppauge, NY), followed by resuspension of pellets in 70 μL of DI water to eliminate any interference from materials in supernatant during assays.Filtration efficiency testsThe filters were exposed to the virus aerosols at 3, 10 and 17 kPa and suspensions of the filters were obtained, as described above. The filtration efficiency was calculated as the ratio of the amount of virus (i.e., total proteins measured from the virus) reconstituted from the filter to that from the virus in the exposure aerosols. The concentration of virus in aerosols was determined by generating viral aerosols into a 15 mL centrifuge tube, containing 1 mL of DI water. After vortexing, virus concentrations (i.e., total protein concentration) were measured with bicinchoninic acid assay (BCA protein assay kit; Thermo Fischer scientific, Waltham, IL) with bovine serum albumin as a standard. In the case of virus reconstituted from salt-coated filters, virus-laden filter suspension was replaced with DI water prior to BCA assay.In vivo infection testsLethal infectivity of influenza viruses (CA/09 H1N1) was examined in 8 week old female inbred BALB/c mice (Nara Biotech; Seoul, Korea) by using the intranasal route. For bare and salt-coated filters, 12 mice per group were infected with individual penetration dosage of influenza virus through each filter. The penetration dosage of the virus through the filters (Filterbare, Filterwet, Filterwet+600μL, and Filterwet+1200μL) was calculated from the filtration efficiency at 10 kPa (near breathing pressure) using the relationship: penetration dosage = virus dosage in lethal aerosol × penetration efficiency (%)/100, where penetration efficiency (%) = 100 − filtration efficiency (%). To examine the effects of the aerosolization process on the viral infectivity change, two mice groups were infected with a lethal dose of virus before and after aerosol formation, which served as negative control groups. Body weight changes and survival rate of mice were monitored daily for 15 days. Mice with body weight loss greater than 25% were euthanized. All animal protocols were approved by the Kyung Hee University (KHU) Institutional Animal Care and Use Committee (IACUC). All animal experiments and husbandry involved in this work were conducted under the approved protocols and guidelines of KHU IACUC. KHU IACUC operates under National Veterinary Research and Quarantine Service (NVRQS), and animal welfare law and regulations of the WOAH-OIE (World organization for animal health).To test strain-dependent lethal infection behavior, mice (12 per group) were infected with the penetrated dosage of viral aerosols (PR/34 H1N1 and VN/04 H5N1 viruses) through Filterwet+600μL at 10 kPa. Time-dependent body weight change was monitored in the same manner described above.Lung viral titer and lung inflammatory cytokine assays after infectionOn day 4 after infection 6 mice of each group were sacrificed for the collection of lung samples. Lung virus titers were measured on six-well plates containing confluent MDCK cell monolayers. Inflammatory cytokines (IFN-γ) were determined using BD OptEIA mouse IFN-γ ELISA kit (BD Biosciences, San Jose, CA) following the manufacturer’s procedure.Test of viral infectivity change on filtersTo investigate the effects of salt-coating on viral infectivity loss, lethal influenza aerosols were exposed to four different types of filters (Filterbare, Filterwet, Filterwet+600μL, and Filterwet+1200μL). Since Filterbare exhibited almost complete penetration upon pressure application, aerosols were exposed to the bare filter in the absence of pressure and samples were carefully handled to prevent mechanical agitation. To measure time-dependent stability change of virus, virus-laden filters were incubated at ambient conditions for 0, 5, 15, and 60 min after aerosol exposure, and suspended in DI water to reconstitute virus at each time point. In vitro stability of virus was characterized by measuring hemagglutinin activity (HA) and virus titers at the same concentration as lethal dose30. The conformational stability of antigenic proteins was characterized by measuring intrinsic fluorescence using 0.1 mg/mL of virus suspension38. To investigate morphological change of virus, lipid stability of viral wall was characterized by nile red fluorescence (Sigma Aldrich), a fluorescent lipid stain, following manufacturer’s protocol39. A decrease in fluorescence intensity can be used to examine the level of disintegration of the virus. Both intrinsic and nile red fluorescence were measured by using a fluorimeter (LB 50B; PerkinElmer, Waltham, MA). Intensity changes of fluorescent spectra were compared relative to those of a control from virus stock.To test infectivity difference observed from in vitro findings, in vivo study was performed for the virus reconstituted from the filters (Filterbare, Filterwet, Filterwet+600μL, and Filterwet+1200μL) after incubation for 60 min at RT (aerosol exposure at 10 kPa, except for Filterbare). 12 mice per group were infected with a lethal dose of virus collected from each type of filter. Body weight change and lung virus titers were measured as described above.Effects of environmental conditions on the performance of salt-coated filterSalt-coated filters (Filterwet, Filterwet+600μL, and Filterwet+1200μL) were stored at 37 °C, 70% RH in an incubator (Maru Max; Rcom, Gyeonggi-do, South Korea) for 15 days. Every day, the filters were collected and incubated at ambient conditions for 5 min. At 1-day incubation, filtration efficiency was measured at 10 kPa from Filterwet+600μL, followed by in vivo infection test. Lethal infectivity between two different filter groups (before and after incubation at 37 °C, 70% RH) was compared by measuring body weight change and survival rate of mice after exposure to lethal CA/09 H1N1 aerosols. XRD analysis was performed to salt-coated filters incubated for 1 and 15 days, and SEM/EDX mapping analysis for 15-day incubated samples.Contact angle measurements and imaging of aerosolsThe bare and salt-coated filters were fixed with carbon tape (Ted Pella, Inc., Redding, CA) to a metal, flat substrate and 3 μL of DI water were added on the surface of the filters. The contact angles were measured from images collected with an optical microscope (10× lens, Motic SMZ-140; Motic, Richmond, Canada) at RT. Images of aerosols on filter fibers were obtained using a dispersive Raman microscope (Nicolet Almega XR; Fisher Scientific).Aerosol drying time on filtersThe bare and salt-coated filters were fixed with carbon tape to a metal, flat substrate and exposed to aerosols generated from 5 μL of Sulforhodamine B Dye solution (1 mM, Sigma-Aldrich). Aerosol drying time was determined with timer by observation with optical microscope.Electron microscopy analysisFor virus stability tests, bare and salt-coated filters were exposed to CA/09 H1N1 aerosols and, after 5 and 60 min incubation, virus was recovered by suspension of the filters, as described above. To study the effects of the coating formulation during aerosol drying independently from crystal growth, bare and salt-coated filters were immersed in DI water and removed after 60 min. Subsequently, virus was incubated in the obtained suspension for 60 min. Additionally, the virus suspension was centrifuged at 19,800 g and 4 °C for 10 min to collect the samples and suspend them in DI water. For TEM analysis (200 kV, JEOL JEM 2100; JEOL, Peabody, MA), samples were deposited on copper grid (Electron Microscopy Sciences, Hatfield, PA) and negatively stained with solution comprised of phosphotungstic acid hydrate (1.5 w/v%, pH = 7.0; Sigma-Aldrich, Oakville, Canada).To identify the morphology of salt-coated filters and recrystallized salts, SEM/EDX analysis was performed for bare and salt-coated filters after coating with 10 nm thick gold layer. Scanning electron microscopy analysis (Hitachi S-3000N; Hitachi, Toronto, Canada) was operated in secondary electron mode at 20 kV and EDX analysis was obtained with EDX detector (Oxford Instruments, Concord, MA).XRD analysisTo confirm the formation of crystalline NaCl coating during drying process and its stability during storage at 37 °C and 70% RH, XRD analysis (BRU-1098; Bruker, Billerica, MA) was performed at different coating conditions. Filters (1 × 1 cm) were mounted on a slide glass for XRD analysis (θ–2θ mode) using a CuKα radiation.Statistical analysisTo compare multiple conditions, Student’s t-test, One-way analysis of variance (ANOVA), and general linear model were used (Minitab release 14; Minitab, State College, PA). P value of less than 0.05 was considered to be significant.Additional InformationHow to cite this article: Quan, F.-S. et al. Universal and reusable virus deactivation system for respiratory protection. Sci. Rep. 7, 39956; doi: 10.1038/srep39956 (2017).Publisher's note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.\n \n\n References1Tellier, R. Review of aerosol transmission of influenza A virus. Emerging Infect. Dis. 12, 1657–1662 (2006).2Weber, T. P. & Stilianakis, N. I. Inactivation of influenza A viruses in the environment and modes of transmission: a critical review. J. Infect. 57, 361–373 (2008).3Duy, J. et al. Circulating microRNA profiles of Ebola virus infection. Sci. Rep. 6, 24496 (2016).4Morse, S. S., Garwin, R. L. & Olsiewski, P. J. Next flu pandemic: what to do until the vaccine arrives? Science 314, 929–929 (2006).5CDC. Laboratory performance evaluation of N95 filtering facepiece respirators, 1996. MMWR, Morbidity and Mortality Weekly Report 47, 1045 (1998).6Seto, W. H. et al. Effectiveness of precautions against droplets and contact in prevention of nosocomial transmission of severe acute respiratory syndrome (SARS). Lancet 361, 1519–1520 (2003).7Bałazy, A. et al. Do N95 respirators provide 95% protection level against airborne viruses, and how adequate are surgical masks? Am. J. Infect. Control 34, 51–57 (2006).8Loeb, M. et al. Surgical mask vs N95 respirator for preventing influenza among health care workers: a randomized trial. JAMA 302, 1865–1871 (2009).9Bunyan, D., Ritchie, L., Jenkins, D. & Coia, J. E. Respiratory and facial protection: a critical review of recent literature. J. Hosp. Infect. 85, 165–169 (2013).10Zhu, C., Lin, C. H. & Cheung, C. S. Inertial impaction-dominated fibrous filtration with rectangular or cylindrical fibers. Powder Technol. 112, 149–162 (2000).11Lee, K. W. & Liu, B. Y. H. Theoretical study of aerosol filtration by fibrous filters. Aerosol Sci. Tech. 1, 147–161 (1982).12Duarte Fo, O. B., Marra, W. D., Kachan, G. C. & Coury, J. R. Filtration of electrified solid particles. Ind. Eng. Chem. Res 39, 3884–3895 (2000).13Kraemer, H. F. & Johnstone, H. F. Collection of aerosol particles in presence of electrostatic fields. Ind. Eng. Chem. 47, 2426–2434 (1955).14Ranz, W. E. & Wong, J. B. Impaction of dust and smoke particles on surface and body collectors. Ind. Eng. Chem. 44, 1371–1381 (1952).15Siegel, J. D., Rhinehart, E., Jackson, M. & Chiarello, L. 2007 guideline for isolation precautions: preventing transmission of infectious agents in health care settings. Am. J. Infect. Control 35, S65–S164 (2007).16Coia, J. E. et al. Guidance on the use of respiratory and facial protection equipment. J. Hosp. Infect. 85, 170–182 (2013).17Ratnesar-Shumate, S. et al. Evaluation of physical capture efficiency and disinfection capability of an iodinated biocidal filter medium. Aerosol Air Qual. Res. 8, 1–18 (2008).18Lee, J. H., Wu, C. Y., Wysocki, K. M. & Farrah, S. & Wander, J. Efficacy of iodine-treated biocidal filter media against bacterial spore aerosols. J. Appl. Microbiol. 105, 1318–1326 (2008).19Badrossamay, M. R. & Sun, G. Acyclic halamine polypropylene polymer: effect of monomer structure on grafting efficiency, stability and biocidal activities. React. Funct. Polym. 68, 1636–1645 (2008).20Zhao, N. & Liu, S. Thermoplastic semi-IPN of polypropylene (PP) and polymeric N-halamine for efficient and durable antibacterial activity. Eur. Polym. J. 47, 1654–1663 (2011).21Cerkez, I., Worley, S. D., Broughton, R. M. & Huang, T. S. Antimicrobial surface coatings for polypropylene nonwoven fabrics. React. Funct. Polym. 73, 1412–1419 (2013).22Davison, A. M. Pathogen inactivation and filtration efficacy of a new anti-microbial and anti-viral surgical facemask and N95 against dentistry-associated microorganisms. International dentistry Australasian edition 7, 36–42 (2012).23Borkow, G., Zhou, S. S., Page, T. & Gabbay, J. A novel anti-influenza copper oxide containing respiratory face mask. PLoS One 5, e11295 (2010).24Borkow, G. & Gabbay, J. Putting copper into action: copper-impregnated products with potent biocidal activities. FASEB J. 18, 1728–1730 (2004).25Li, Y., Leung, P., Yao, L., Song, Q. W. & Newton, E. Antimicrobial effect of surgical masks coated with nanoparticles. J. Hosp. Infect. 62, 58–63 (2006).26Lee, J. H. et al. Assessment of iodine-treated filter media for removal and inactivation of MS2 bacteriophage aerosols. J. Appl. Microbiol. 107, 1912–1923 (2009).27Lore, M. B. et al. Performance of conventional and antimicrobial-treated filtering facepiece respirators challenged with biological aerosols. J. Occup. Environ. Hyg. 9, 69–80 (2012).28Rengasamy, S., Fisher, E. & Shaffer, R. E. Evaluation of the survivability of MS2 viral aerosols deposited on filtering face piece respirator samples incorporating antimicrobial technologies. Am. J. Infect. Control 38, 9–17 (2010).29Windisch, W., Hennings, E., Sorichter, S., Hamm, H. & Criee, C. P. Peak or plateau maximal inspiratory mouth pressure: which is best? Eur. Respir. J. 23, 708–713 (2004).30Choi, H.-J. et al. Stability of influenza vaccine coated onto microneedles. Biomaterials 33, 3756–3769 (2012).31Choi, H.-J. et al. Stability of whole inactivated influenza virus vaccine during coating onto metal microneedles. J. Control. Release 166, 159–171 (2013).32Baldwin, R. L. How hofmeister ion interactions affect protein stability. Biophys. J. 71, 2056–2063 (1996).33Choi, H.-J. et al. Effect of osmotic pressure on the stability of whole inactivated influenza vaccine for coating on microneedles. PLoS one 10, e0134431 (2015).34ASTM International. ASTM F2101 - 14 Standard Test Method for Evaluating the Bacterial Filtration Efficiency (BFE) of Medical Face Mask Materials, Using a Biological Aerosol of Staphylococcus aureus (2014).35Adams, J. R. & Merz, A. R. Hygroscopicity of fertilizer materials and mixtures. Ind. Eng. Chem. Res. 21, 305–307 (1929).36Song, J.-M. et al. Protective immunity against H5N1 influenza virus by a single dose vaccination with virus-like particles. Virology 405, 165–175 (2010).37Quan, F. S. et al. A bivalent influenza VLP vaccine confers complete inhibition of virus replication in lungs. Vaccine 26, 3352–3361 (2008).38Choi, H.-J., Ebersbacher, C. F., Kim, M., Kang, S. & Montemagno, C. D. A mechanistic study on the destabilization of whole inactivated influenza virus vaccine in gastric environment. PLoS one 8, e66316 (2013).39Greenspan, P., Mayer, E. P. & Fowler, S. D. Nile red: a selective fluorescent stain for intracellular lipid droplets. J. Cell Biol. 100, 965–973 (1985).Download referencesAcknowledgementsThis research was financially supported by startup funds from University of Alberta (H.J.C.), and grants from National Research Foundation of Korea (NRF) (NRF-2014R1A2A2A01004899) and Ministry of Health & Welfare, Republic of Korea (HI15C2928).Author informationAuthor notesFu-Shi Quan and Ilaria Rubino: These authors contributed equally to this work.AffiliationsDepartment of Medical Zoology, Kyung Hee University School of Medicine, Seoul, 130-701, KoreaFu-Shi QuanDepartment of Chemical and Materials Engineering, University of Alberta, Edmonton, AB T6G 1H9, CanadaIlaria Rubino, Brendan Koch & Hyo-Jick ChoiDepartment of Biomedical Science, Graduate School, Kyung Hee University, Seoul, 130-701, KoreaSu-Hwa LeeAuthorsSearch for Fu-Shi Quan in:Search for Ilaria Rubino in:Search for Su-Hwa Lee in:Search for Brendan Koch in:Search for Hyo-Jick Choi in:ContributionsH.J.C. conceived and designed the experiments. F.S.Q., I.R., S.H.L., B.K., and H.J.C. performed the experiments. F.S.Q., I.R., S.H.L., B.K., and H.J.C. analyzed the data. I.R. and H.J.C. wrote the manuscript. F.S.Q. and B.K. edited the manuscript.Corresponding authorCorrespondence to\n Hyo-Jick Choi.Ethics declarations\n \n Competing interests\n The authors declare no competing financial interests.\n \n Supplementary informationRights and permissionsThis work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/Reprints and PermissionsAbout this articleCite this articleQuan, F., Rubino, I., Lee, S. et al. Universal and reusable virus deactivation system for respiratory protection.\n Sci Rep 7, 39956 (2017). https://doi.org/10.1038/srep39956Download citationReceived: 04 August 2016Accepted: 30 November 2016Published: 04 January 2017DOI: https://doi.org/10.1038/srep39956\n\n \n \n \n Further reading\n \n \n \n \n \n \n Survival of Microorganisms on Nonwovens Used for the Construction of Filtering Facepiece Respirators\n \n \n \n \n Katarzyna Majchrzycka, Małgorzata Okrasa, Justyna Szulc, Anita Jachowicz[…] & Beata Gutarowska- Show fewer authors\n \n \n International Journal of Environmental Research and Public Health\n (2019)\n \n \n \n \n \n \n Respiratory Protection against Pandemic and Epidemic Diseases\n \n \n \n \n Ilaria Rubino & Hyo-Jick Choi\n \n \n Trends in Biotechnology\n (2017)\n \n \n \n \n \n \n \n \n\n \n CommentsBy submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate."],["topImageUrl",null],["error",null]]}]}],["text","WIKILINK\n最新科技資訊\n\n \n口罩能否反復使用？浸鹽水後可100%讓H1N1病毒滅活，但對其他病毒需謹慎 | 《科學報告》期刊\nFebruary 5, 2020 admin\n曉查發自凹非寺 \n量子位報道 | 公眾號 QbitAI\n一場新冠肺炎讓中國的口罩嚴重告急。\n\n每齣一次門就要廢掉一個口罩，傢裏的口罩即將用完，藥店和網上都買不到，真是急死人瞭。\n\n彆怕，早在2016年，來自韓國慶熙大學的研究者就發現，口罩隻需用普通食鹽水處理後，就具有瞭一定的殺滅病毒能力，至少對2009年爆發的H1N1流感病毒有效。\n\n他們將處理後的口罩來過濾充滿H1N1病毒的空氣，然後再給小白鼠呼吸，結果發現小鼠的存活率達到瞭100%。\n\n\n\n\n \n空氣中的病毒濃度達到瞭緻死量。作為對照，如果使用普通醫用口罩過濾，小白鼠在4天後就會齣現嚴重的肺炎，10天後全部死亡。\n\n這篇文章發錶在2017年1月Nature旗下的Scientific Reports期刊上。\n\n\n\n△實驗中用到的過濾裝置\n早在3年前，該文章就預測瞭口罩短缺的可能性。\n\n作者在開頭說，在疫病大流行時期，生産能力不足和製造過程耗時，會導緻口罩供應有限。\n\nN95口罩雖然可以過濾空氣中形成的病毒氣溶膠，但是在嚴重的疫情麵前，大規模使用N95口罩是不切實際的，而且價格昂貴。\n\n過去的SARS、H1N1、MERS都錶明，醫用外科口罩是被公眾最廣泛采用的個人防護措施，盡管人們對它的效果存在爭議。\n\n因此需要開發一種通用、低成本且有效的防護病毒機製，最好能用在普通醫用口罩上。\n\n\n\n\n\n\n研究人員配置瞭濃度29.03 w/v%的氯化鈉溶液，並在其中加入瞭一些Tween 20錶麵活性劑，將口罩浸潤，然後在37℃的環境下乾燥一天。\n\n文章指齣，經過這種方法處理後的口罩無需再次處理即可重復使用。最終實驗發現，塗有氯化鈉的口罩上的病毒在5分鍾後就失去瞭活性。\n\n\n\n\n\n\n從顯微鏡分析來看，病毒氣溶膠乾燥時間約為3分鍾，錶明在5分鍾時觀察到的病毒破壞與乾燥引起的鹽結晶有關。\n\n作者推測，可能是帶有水分的病毒被吸附在口罩錶麵，濕潤後又重結晶的鹽破壞瞭病毒的結構。\n\n既然對H1N1病毒，自然就有網友很好奇，這種處理方法是否也對新型冠狀病毒有效。\n\n一些網友對此提齣瞭質疑，認為鹽結晶可能減小瞭口罩的通氣量，而且實驗隻對H1N1病毒進行瞭驗證，是否實用還有待商榷。\n\n所以在未經實驗驗證的情況下，請不要用這種方法來過濾新型冠狀病毒。口罩還是要保證一用一換，切不可自行處理後就重復使用，否則仍有被感染的風險。\n\n論文地址：\nhttps://www.nature.com/articles/srep39956#Sec8\n\n— 完 —\n\n"],["id","obwrpyiajxnx"],["createdAt","2020-02-09T01:53:22.130Z"]]}],["reply",{"__iterable":"Map","data":[["id","tmgTKHABd3n3h-WYriSs"],["user",{"__iterable":"Map","data":[["id","_1YnAG8BV0DITCqGm7go"],["name","uienwt"],["avatarUrl",null],["level",18]]}],["type","OPINIONATED"],["text","目前沒有證據顯示鹽水有助於消除新型冠狀病毒。用鹽殺菌的原理是應用在醃漬食物，讓細菌無法生長，但人不可能用高濃度的鹽水來浸泡口罩，因此傳言指稱用淡鹽水漱口是沒有效果的。"],["reference","https://tfc-taiwan.org.tw/articles/2119\n"],["hyperlinks",{"__iterable":"List","data":[{"__iterable":"Map","data":[["title","【錯誤】網傳「鐘院士的防病毒高招：建議各位去醫院或其他公共場合之前用淡鹽水漱一下嚥喉部位，回家後再漱一次...」？"],["url","https://tfc-taiwan.org.tw/articles/2119"],["summary","【錯誤】網傳「鐘院士的防病毒高招：建議各位去醫院或其他公共場合之前用淡鹽水漱一下嚥喉部位，回家後再漱一次...」？\n \n \n\n網傳「鐘院士的防病毒高招：建議各位去醫院或其他公共場合之前用淡鹽水漱一下嚥喉部位，回家後再漱一次...」，經查：\n\n一、中國工程院院士、中國「新型冠狀病毒聯防聯控工作機制科研攻關組」組長鐘南山的團隊指出，此傳言為網路謠言。\n\n二、專家指出，鹽水漱口不能預防新型冠狀病毒。\n\n因此，此傳言為「錯誤」訊息。\n\n背景\n\n社群平台、通訊軟體流傳訊息指出：「鐘院士的防病毒高招：建議各位去醫院或其他公共場合之前用淡鹽水漱一下嚥喉部位，回家後再漱一次...由於病毒或細菌首先通過鼻腔潛伏於咽部，淡鹽水可以在第一時間殺滅它們，從而達到預防感染的目的」？\n\n\n\n圖1：社群平台流傳訊息擷圖\n\n查核\n\n爭議點一、鐘南山是否有發出此警告？\n\n鐘南山是衛生領域的中國工程院院士，日前接任中國「新型冠狀病毒聯防聯控工作機制科研攻關組」的組長，是中國2003年抗SARS的重要專家。鐘南山團隊所在的廣州醫科大學附屬第一醫院官方微博帳號，於2020年1月22日發布闢謠貼文，指出此貼文為網路謠言。\n\n\n\n圖2：鐘南山團隊發布闢謠貼文擷圖\n\n爭議點二、「用鹽水漱口」可以預防武漢肺炎嗎？傳言指稱「由於病毒或細菌首先通過鼻腔潛伏於咽部，淡鹽水可以在第一時間殺滅它們，從而達到預防感染的目的」，是否屬實？\n\n鐘南山團隊所在的廣州醫科大學附屬第一醫院官方微博指出，鹽水漱口有助於清潔口腔和咽喉，但新型冠狀病毒侵犯的是呼吸道，漱口並沒有辦法清潔呼吸道。目前也沒有證據顯示鹽水有助於消除新型冠狀病毒。\n\n查核中心透過新興科技媒體中心，採訪中興大學獸醫病理生物學研究所教授吳弘毅。他表示，用鹽殺菌的原理是應用在醃漬食物，讓細菌無法生長，但人不可能用高濃度的鹽水來清洗鼻腔或咽喉，因此傳言指稱用淡鹽水漱口是沒有效果的。此外，生理食鹽水反而是病毒可以停留比較久的環境。\n\n台灣感染症醫學會理事長、台大醫院小兒部主任黃立民說，病毒很快就會進入細胞，所以去用鹽水清洗鼻腔或咽喉，並沒有任何作用，而且，鹽水也沒有消毒殺菌的作用。\n\n查核中心採訪長庚大學新興病毒感染研究中心主任施信如，她說此種方法沒有科學根據，也沒有辦法達到預防感染的效果。\n\n結論\n\n一、中國工程院院士、中國「新型冠狀病毒聯防聯控工作機制科研攻關組」組長鐘南山的團隊指出，此傳言為網路謠言。\n\n二、專家指出，鹽水漱口不能預防新型冠狀病毒。\n\n因此，此傳言為「錯誤」訊息。\n\n參考資料\n\n廣州醫科大學附屬第一醫院官方微博"],["topImageUrl","https://tfc-taiwan.org.tw/sites/default/files/upload/node/field_blog_images/image1_2.png"],["error",null]]}]}],["createdAt","2020-02-09T03:52:23.478Z"]]}]]},{"__iterable":"Map","data":[["article",{"__iterable":"Map","data":[["replyCount",1],["references",{"__iterable":"List","data":[{"__iterable":"Map","data":[["type","LINE"]]}]}],["replyRequestCount",1],["user",{"__iterable":"Map","data":[["id","j4S8C_n1X_AVJRpVDJaAWXXARts8Z96DWjC7LTqy8kpttWQvw"],["name","☞健談的坪林韋納爾☜"],["avatarUrl",null],["level",0]]}],["hyperlinks",{"__iterable":"List","data":[{"__iterable":"Map","data":[["title","Clinical features of patients infected with 2019 novel coronavirus in Wuhan, China"],["url","https://www.thelancet.com/journals/lancet/article/PIIS0140-6736(20)30183-5/fulltext"],["summary","SummaryBackgroundA recent cluster of pneumonia cases in Wuhan, China, was caused by a novel betacoronavirus, the 2019 novel coronavirus (2019-nCoV). We report the epidemiological, clinical, laboratory, and radiological characteristics and treatment and clinical outcomes of these patients.MethodsAll patients with suspected 2019-nCoV were admitted to a designated hospital in Wuhan. We prospectively collected and analysed data on patients with laboratory-confirmed 2019-nCoV infection by real-time RT-PCR and next-generation sequencing. Data were obtained with standardised data collection forms shared by the International Severe Acute Respiratory and Emerging Infection Consortium from electronic medical records. Researchers also directly communicated with patients or their families to ascertain epidemiological and symptom data. Outcomes were also compared between patients who had been admitted to the intensive care unit (ICU) and those who had not.FindingsBy Jan 2, 2020, 41 admitted hospital patients had been identified as having laboratory-confirmed 2019-nCoV infection. Most of the infected patients were men (30 [73%] of 41); less than half had underlying diseases (13 [32%]), including diabetes (eight [20%]), hypertension (six [15%]), and cardiovascular disease (six [15%]). Median age was 49·0 years (IQR 41·0–58·0). 27 (66%) of 41 patients had been exposed to Huanan seafood market. One family cluster was found. Common symptoms at onset of illness were fever (40 [98%] of 41 patients), cough (31 [76%]), and myalgia or fatigue (18 [44%]); less common symptoms were sputum production (11 [28%] of 39), headache (three [8%] of 38), haemoptysis (two [5%] of 39), and diarrhoea (one [3%] of 38). Dyspnoea developed in 22 (55%) of 40 patients (median time from illness onset to dyspnoea 8·0 days [IQR 5·0–13·0]). 26 (63%) of 41 patients had lymphopenia. All 41 patients had pneumonia with abnormal findings on chest CT. Complications included acute respiratory distress syndrome (12 [29%]), RNAaemia (six [15%]), acute cardiac injury (five [12%]) and secondary infection (four [10%]). 13 (32%) patients were admitted to an ICU and six (15%) died. Compared with non-ICU patients, ICU patients had higher plasma levels of IL2, IL7, IL10, GSCF, IP10, MCP1, MIP1A, and TNFα.InterpretationThe 2019-nCoV infection caused clusters of severe respiratory illness similar to severe acute respiratory syndrome coronavirus and was associated with ICU admission and high mortality. Major gaps in our knowledge of the origin, epidemiology, duration of human transmission, and clinical spectrum of disease need fulfilment by future studies.FundingMinistry of Science and Technology, Chinese Academy of Medical Sciences, National Natural Science Foundation of China, and Beijing Municipal Science and Technology Commission.IntroductionCoronaviruses are enveloped non-segmented positive-sense RNA viruses belonging to the family Coronaviridae and the order Nidovirales and broadly distributed in humans and other mammals.1Richman DD Whitley RJ Hayden FG Although most human coronavirus infections are mild, the epidemics of the two betacoronaviruses, severe acute respiratory syndrome coronavirus (SARS-CoV)2Ksiazek TG Erdman D Goldsmith CS et al.A novel coronavirus associated with severe acute respiratory syndrome., 3Kuiken T Fouchier RAM Schutten M et al.Newly discovered coronavirus as the primary cause of severe acute respiratory syndrome., 4Drosten C Günther S Preiser W et al.Identification of a novel coronavirus in patients with severe acute respiratory syndrome. and Middle East respiratory syndrome coronavirus (MERS-CoV),5de Groot RJ Baker SC Baric RS et al.Middle East respiratory syndrome coronavirus (MERS-CoV): announcement of the Coronavirus Study Group., 6Zaki AM van Boheemen S Bestebroer TM Osterhaus ADME Fouchier RAM Isolation of a novel coronavirus from a man with pneumonia in Saudi Arabia. have caused more than 10 000 cumulative cases in the past two decades, with mortality rates of 10% for SARS-CoV and 37% for MERS-CoV.7WHOSummary of probable SARS cases with onset of illness from 1 November 2002 to 31 July 2003., 8WHOMiddle East respiratory syndrome coronavirus (MERS-CoV). The coronaviruses already identified might only be the tip of the iceberg, with potentially more novel and severe zoonotic events to be revealed.In December, 2019, a series of pneumonia cases of unknown cause emerged in Wuhan, Hubei, China, with clinical presentations greatly resembling viral pneumonia.9WHONovel coronavirus – China. Deep sequencing analysis from lower respiratory tract samples indicated a novel coronavirus, which was named 2019 novel coronavirus (2019-nCoV). Thus far, more than 800 confirmed cases, including in health-care workers, have been identified in Wuhan, and several exported cases have been confirmed in other provinces in China, and in Thailand, Japan, South Korea, and the USA.10WHONovel coronavirus – Thailand (ex-China)., 11WHONovel coronavirus – Japan (ex-China)., 12WHONovel coronavirus – Republic of Korea (ex-China)., 13CDCFirst travel-related case of 2019 novel coronavirus detected in United States.We aim to describe epidemiological, clinical, laboratory, and radiological characteristics, treatment, and outcomes of patients confirmed to have 2019-nCoV infection, and to compare the clinical features between intensive care unit (ICU) and non-ICU patients. We hope our study findings will inform the global community of the emergence of this novel coronavirus and its clinical features.Methods PatientsFollowing the pneumonia cases of unknown cause reported in Wuhan and considering the shared history of exposure to Huanan seafood market across the patients, an epidemiological alert was released by the local health authority on Dec 31, 2019, and the market was shut down on Jan 1, 2020. Meanwhile, 59 suspected cases with fever and dry cough were transferred to a designated hospital starting from Dec 31, 2019. An expert team of physicians, epidemiologists, virologists, and government officials was soon formed after the alert.Since the cause was unknown at the onset of these emerging infections, the diagnosis of pneumonia of unknown cause in Wuhan was based on clinical characteristics, chest imaging, and the ruling out of common bacterial and viral pathogens that cause pneumonia. Suspected patients were isolated using airborne precautions in the designated hospital, Jin Yin-tan Hospital (Wuhan, China), and fit-tested N95 masks and airborne precautions for aerosol-generating procedures were taken. This study was approved by the National Health Commission of China and Ethics Commission of Jin Yin-tan Hospital (KY-2020-01.01). Written informed consent was waived by the Ethics Commission of the designated hospital for emerging infectious diseases. ProceduresLocal centres for disease control and prevention collected respiratory, blood, and faeces specimens, then shipped them to designated authoritative laboratories to detect the pathogen (NHC Key Laboratory of Systems Biology of Pathogens and Christophe Mérieux Laboratory, Beijing, China). A novel coronavirus, which was named 2019-nCoV, was isolated then from lower respiratory tract specimen and a diagnostic test for this virus was developed soon after that.14Tan W Zhao X Ma X et al.A novel coronavirus genome identified in a cluster of pneumonia cases — Wuhan, China 2019−2020. Of 59 suspected cases, 41 patients were confirmed to be infected with 2019-nCoV. The presence of 2019-nCoV in respiratory specimens was detected by next-generation sequencing or real-time RT-PCR methods. The primers and probe target to envelope gene of CoV were used and the sequences were as follows: forward primer 5′-TCAGAATGCCAATCTCCCCAAC-3′; reverse primer 5′-AAAGGTCCACCCGATACATTGA-3′; and the probe 5′CY5-CTAGTTACACTAGCCATCCTTACTGC-3′BHQ1. Conditions for the amplifications were 50°C for 15 min, 95°C for 3 min, followed by 45 cycles of 95°C for 15 s and 60°C for 30 s.Initial investigations included a complete blood count, coagulation profile, and serum biochemical test (including renal and liver function, creatine kinase, lactate dehydrogenase, and electrolytes). Respiratory specimens, including nasal and pharyngeal swabs, bronchoalveolar lavage fluid, sputum, or bronchial aspirates were tested for common viruses, including influenza, avian influenza, respiratory syncytial virus, adenovirus, parainfluenza virus, SARS-CoV and MERS-CoV using real-time RT-PCR assays approved by the China Food and Drug Administration. Routine bacterial and fungal examinations were also performed.Given the emergence of the 2019-nCoV pneumonia cases during the influenza season, antibiotics (oral and intravenous) and oseltamivir (orally 75 mg twice daily) were empirically administered. Corticosteroid therapy (methylprednisolone 40–120 mg per day) was given as a combined regimen if severe community-acquired pneumonia was diagnosed by physicians at the designated hospital. Oxygen support (eg, nasal cannula and invasive mechanical ventilation) was administered to patients according to the severity of hypoxaemia. Repeated tests for 2019-nCoV were done in patients confirmed to have 2019-nCoV infection to show viral clearance before hospital discharge or discontinuation of isolation. Data collectionWe reviewed clinical charts, nursing records, laboratory findings, and chest x-rays for all patients with laboratory-confirmed 2019-nCoV infection who were reported by the local health authority. The admission data of these patients was from Dec 16, 2019, to Jan 2, 2020. Epidemiological, clinical, laboratory, and radiological characteristics and treatment and outcomes data were obtained with standardised data collection forms (modified case record form for severe acute respiratory infection clinical characterisation shared by the International Severe Acute Respiratory and Emerging Infection Consortium) from electronic medical records. Two researchers also independently reviewed the data collection forms to double check the data collected. To ascertain the epidemiological and symptom data, which were not available from electronic medical records, the researchers also directly communicated with patients or their families to ascertain epidemiological and symptom data. Cytokine and chemokine measurementTo characterise the effect of coronavirus on the production of cytokines or chemokines in the acute phase of the illness, plasma cytokines and chemokines (IL1B, IL1RA, IL2, IL4, IL5, IL6, IL7, IL8 (also known as CXCL8), IL9, IL10, IL12p70, IL13, IL15, IL17A, Eotaxin (also known as CCL11), basic FGF2, GCSF (CSF3), GMCSF (CSF2), IFNγ, IP10 (CXCL10), MCP1 (CCL2), MIP1A (CCL3), MIP1B (CCL4), PDGFB, RANTES (CCL5), TNFα, and VEGFA were measured using Human Cytokine Standard 27-Plex Assays panel and the Bio-Plex 200 system (Bio-Rad, Hercules, CA, USA) for all patients according to the manufacturer's instructions. The plasma samples from four healthy adults were used as controls for cross-comparison. The median time from being transferred to a designated hospital to the blood sample collection was 4 days (IQR 2–5). Detection of coronavirus in plasmaEach 80 μL plasma sample from the patients and contacts was added into 240 μL of Trizol LS (10296028; Thermo Fisher Scientific, Carlsbad, CA, USA) in the Biosafety Level 3 laboratory. Total RNA was extracted by Direct-zol RNA Miniprep kit (R2050; Zymo research, Irvine, CA, USA) according to the manufacturer's instructions and 50 μL elution was obtained for each sample. 5 μL RNA was used for real-time RT-PCR, which targeted the NP gene using AgPath-ID One-Step RT-PCR Reagent (AM1005; Thermo Fisher Scientific). The final reaction mix concentration of the primers was 500 nM and probe was 200 nM. Real-time RT-PCR was performed using the following conditions: 50°C for 15 min and 95°C for 3 min, 50 cycles of amplification at 95°C for 10 s and 60°C for 45 s. Since we did not perform tests for detecting infectious virus in blood, we avoided the term viraemia and used RNAaemia instead. RNAaemia was defined as a positive result for real-time RT-PCR in the plasma sample. DefinitionsAcute respiratory distress syndrome (ARDS) and shock were defined according to the interim guidance of WHO for novel coronavirus.9WHONovel coronavirus – China. Hypoxaemia was defined as arterial oxygen tension (PaO2) over inspiratory oxygen fraction (FIO2) of less than 300 mm Hg.15Sanz F Gimeno C Lloret T et al.Relationship between the presence of hypoxemia and the inflammatory response measured by C-reactive protein in bacteremic pneumococcal pneumonia. Acute kidney injury was identified and classified on the basis of the highest serum creatinine level or urine output criteria according to the kidney disease improving global outcomes classification.16Kidney disease: improving global outcomes (KDIGO) acute kidney injury work groupKDIGO clinical practice guideline for acute kidney injury. Secondary infection was diagnosed if the patients had clinical symptoms or signs of nosocomial pneumonia or bacteraemia, and was combined with a positive culture of a new pathogen from a lower respiratory tract specimen (including the sputum, transtracheal aspirates, or bronchoalveolar lavage fluid, or from blood samples taken ≥48 h after admission).17Garner JS Jarvis WR Emori TG Horan TC Hughes JM CDC definitions for nosocomial infections, 1988. Cardiac injury followed the definition used in our previous study in H7N9 patients.18Gao C Wang Y Gu X et al.Association between cardiac injury and mortality in hospitalized patients infected with avian influenza A (H7N9) virus. In brief, cardiac injury was diagnosed if serum levels of cardiac biomarkers (eg, troponin I) were above the 99th percentile upper reference limit, or new abnormalities were shown in electrocardiography and echocardiography. Statistical analysisContinuous variables were expressed as median (IQR) and compared with the Mann-Whitney U test; categorical variables were expressed as number (%) and compared by χ2 test or Fisher's exact test between ICU care and no ICU care groups. Boxplots were drawn to describe plasma cytokine and chemokine concentrations.A two-sided α of less than 0·05 was considered statistically significant. Statistical analyses were done using the SAS software, version 9.4, unless otherwise indicated. Role of the funding sourceThe funder of the study had no role in study design, data collection, data analysis, data interpretation, or writing of the report. The corresponding authors had full access to all the data in the study and had final responsibility for the decision to submit for publication.ResultsBy Jan 2, 2020, 41 admitted hospital patients were identified as laboratory-confirmed 2019-nCoV infection in Wuhan. 20 [49%]) of the 2019-nCoV-infected patients were aged 25–49 years, and 14 (34%) were aged 50–64 years (figure 1A). The median age of the patients was 49·0 years (IQR 41·0–58·0; table 1). In our cohort of the first 41 patients as of Jan 2, no children or adolescents were infected. Of the 41 patients, 13 (32%) were admitted to the ICU because they required high-flow nasal cannula or higher-level oxygen support measures to correct hypoxaemia. Most of the infected patients were men (30 [73%]); less than half had underlying diseases (13 [32%]), including diabetes (eight [20%]), hypertension (six [15%]), and cardiovascular disease (six [15%]).Table 1Demographics and baseline characteristics of patients infected with 2019-nCoVData are median (IQR), n (%), or n/N (%), where N is the total number of patients with available data. p values comparing ICU care and no ICU care are from χ2 test, Fisher's exact test, or Mann-Whitney U test. 2019-nCoV=2019 novel coronavirus. ICU=intensive care unit.\n Open table in a new tab\n 27 (66%) patients had direct exposure to Huanan seafood market (figure 1B). Market exposure was similar between the patients with ICU care (nine [69%]) and those with non-ICU care (18 [64%]). The symptom onset date of the first patient identified was Dec 1, 2019. None of his family members developed fever or any respiratory symptoms. No epidemiological link was found between the first patient and later cases. The first fatal case, who had continuous exposure to the market, was admitted to hospital because of a 7-day history of fever, cough, and dyspnoea. 5 days after illness onset, his wife, a 53-year-old woman who had no known history of exposure to the market, also presented with pneumonia and was hospitalised in the isolation ward.The most common symptoms at onset of illness were fever (40 [98%] of 41 patients), cough (31 [76%]), and myalgia or fatigue (18 [44%]); less common symptoms were sputum production (11 [28%] of 39), headache (three [8%] of 38), haemoptysis (two [5%] of 39), and diarrhoea (one [3%] of 38; table 1). More than half of patients (22 [55%] of 40) developed dyspnoea. The median duration from illness onset to dyspnoea was 8·0 days (IQR 5·0–13·0). The median time from onset of symptoms to first hospital admission was 7·0 days (4·0–8·0), to shortness of breath was 8·0 days (5·0–13·0), to ARDS was 9·0 days (8·0–14·0), to mechanical ventilation was 10·5 days (7·0–14·0), and to ICU admission was 10·5 days (8·0–17·0; figure 2).The blood counts of patients on admission showed leucopenia (white blood cell count less than 4 × 109/L; ten [25%] of 40 patients) and lymphopenia (lymphocyte count <1·0 × 109/L; 26 [63%] patients; table 2). Prothrombin time and D-dimer level on admission were higher in ICU patients (median prothrombin time 12·2 s [IQR 11·2–13·4]; median D-dimer level 2·4 mg/L [0·6–14·4]) than non-ICU patients (median prothrombin time 10·7 s [9·8–12·1], p=0·012; median D-dimer level 0·5 mg/L [0·3–0·8], p=0·0042). Levels of aspartate aminotransferase were increased in 15 (37%) of 41 patients, including eight (62%) of 13 ICU patients and seven (25%) of 28 non-ICU patients. Hypersensitive troponin I (hs-cTnI) was increased substantially in five patients, in whom the diagnosis of virus-related cardiac injury was made.Table 2Laboratory findings of patients infected with 2019-nCoV on admission to hospitalData are median (IQR) or n/N (%), where N is the total number of patients with available data. p values comparing ICU care and no ICU care are from χ2, Fisher's exact test, or Mann-Whitney U test. 2019-nCoV=2019 novel coronavirus. ICU=intensive care unit.\n Open table in a new tab\n Most patients had normal serum levels of procalcitonin on admission (procalcitonin <0·1 ng/mL; 27 [69%] patients; table 2). Four ICU patients developed secondary infections. Three of the four patients with secondary infection had procalcitonin greater than 0·5 ng/mL (0·69 ng/mL, 1·46 ng/mL, and 6·48 ng/mL).On admission, abnormalities in chest CT images were detected among all patients. Of the 41 patients, 40 (98%) had bilateral involvement (table 2). The typical findings of chest CT images of ICU patients on admission were bilateral multiple lobular and subsegmental areas of consolidation (figure 3A). The representative chest CT findings of non-ICU patients showed bilateral ground-glass opacity and subsegmental areas of consolidation (figure 3B). Later chest CT images showed bilateral ground-glass opacity, whereas the consolidation had been resolved (figure 3C).Initial plasma IL1B, IL1RA, IL7, IL8, IL9, IL10, basic FGF, GCSF, GMCSF, IFNγ, IP10, MCP1, MIP1A, MIP1B, PDGF, TNFα, and VEGF concentrations were higher in both ICU patients and non-ICU patients than in healthy adults (appendix pp 6–7). Plasma levels of IL5, IL12p70, IL15, Eotaxin, and RANTES were similar between healthy adults and patients infected with 2019-nCoV. Further comparison between ICU and non-ICU patients showed that plasma concentrations of IL2, IL7, IL10, GCSF, IP10, MCP1, MIP1A, and TNFα were higher in ICU patients than non-ICU patients.All patients had pneumonia. Common complications included ARDS (12 [29%] of 41 patients), followed by RNAaemia (six [15%] patients), acute cardiac injury (five [12%] patients), and secondary infection (four [10%] patients; table 3). Invasive mechanical ventilation was required in four (10%) patients, with two of them (5%) had refractory hypoxaemia and received extracorporeal membrane oxygenation as salvage therapy. All patients were administered with empirical antibiotic treatment, and 38 (93%) patients received antiviral therapy (oseltamivir). Additionally, nine (22%) patients were given systematic corticosteroids. A comparison of clinical features between patients who received and did not receive systematic corticosteroids is in the appendix (pp 1–5).Table 3Treatments and outcomes of patients infected with 2019-nCoVData are median (IQR) or n (%). p values are comparing ICU care and no ICU care. 2019-nCoV=2019 novel coronavirus. ICU=intensive care unit. NA=not applicable. ECMO=extracorporeal membrane oxygenation.\n Open table in a new tab\n As of Jan 22, 2020, 28 (68%) of 41 patients have been discharged and six (15%) patients have died. Fitness for discharge was based on abatement of fever for at least 10 days, with improvement of chest radiographic evidence and viral clearance in respiratory samples from upper respiratory tract.DiscussionWe report here a cohort of 41 patients with laboratory-confirmed 2019-nCoV infection. Patients had serious, sometimes fatal, pneumonia and were admitted to the designated hospital in Wuhan, China, by Jan 2, 2020. Clinical presentations greatly resemble SARS-CoV. Patients with severe illness developed ARDS and required ICU admission and oxygen therapy. The time between hospital admission and ARDS was as short as 2 days. At this stage, the mortality rate is high for 2019-nCoV, because six (15%) of 41 patients in this cohort died.The number of deaths is rising quickly. As of Jan 24, 2020, 835 laboratory-confirmed 2019-nCoV infections were reported in China, with 25 fatal cases. Reports have been released of exported cases in many provinces in China, and in other countries; some health-care workers have also been infected in Wuhan. Taken together, evidence so far indicates human transmission for 2019-nCoV. We are concerned that 2019-nCoV could have acquired the ability for efficient human transmission.19Perlman S Netland J Coronaviruses post-SARS: update on replication and pathogenesis. Airborne precautions, such as a fit-tested N95 respirator, and other personal protective equipment are strongly recommended. To prevent further spread of the disease in health-care settings that are caring for patients infected with 2019-nCoV, onset of fever and respiratory symptoms should be closely monitored among health-care workers. Testing of respiratory specimens should be done immediately once a diagnosis is suspected. Serum antibodies should be tested among health-care workers before and after their exposure to 2019-nCoV for identification of asymptomatic infections.Similarities of clinical features between 2019-nCoV and previous betacoronavirus infections have been noted. In this cohort, most patients presented with fever, dry cough, dyspnoea, and bilateral ground-glass opacities on chest CT scans. These features of 2019-nCoV infection bear some resemblance to SARS-CoV and MERS-CoV infections.20Lee N Hui D Wu A et al.A major outbreak of severe acute respiratory syndrome in Hong Kong., 21Assiri A Al-Tawfiq JA Al-Rabeeah AA et al.Epidemiological, demographic, and clinical characteristics of 47 cases of Middle East respiratory syndrome coronavirus disease from Saudi Arabia: a descriptive study. However, few patients with 2019-nCoV infection had prominent upper respiratory tract signs and symptoms (eg, rhinorrhoea, sneezing, or sore throat), indicating that the target cells might be located in the lower airway. Furthermore, 2019-nCoV patients rarely developed intestinal signs and symptoms (eg, diarrhoea), whereas about 20–25% of patients with MERS-CoV or SARS-CoV infection had diarrhoea.21Assiri A Al-Tawfiq JA Al-Rabeeah AA et al.Epidemiological, demographic, and clinical characteristics of 47 cases of Middle East respiratory syndrome coronavirus disease from Saudi Arabia: a descriptive study. Faecal and urine samples should be tested to exclude a potential alternative route of transmission that is unknown at this stage.The pathophysiology of unusually high pathogenicity for SARS-CoV or MERS-CoV has not been completely understood. Early studies have shown that increased amounts of proinflammatory cytokines in serum (eg, IL1B, IL6, IL12, IFNγ, IP10, and MCP1) were associated with pulmonary inflammation and extensive lung damage in SARS patients.22Wong CK Lam CWK Wu AKL et al.Plasma inflammatory cytokines and chemokines in severe acute respiratory syndrome. MERS-CoV infection was also reported to induce increased concentrations of proinflammatory cytokines (IFNγ, TNFα, IL15, and IL17).23Mahallawi WH Khabour OF Zhang Q Makhdoum HM Suliman BA MERS-CoV infection in humans is associated with a pro-inflammatory Th1 and Th17 cytokine profile. We noted that patients infected with 2019-nCoV also had high amounts of IL1B, IFNγ, IP10, and MCP1, probably leading to activated T-helper-1 (Th1) cell responses. Moreover, patients requiring ICU admission had higher concentrations of GCSF, IP10, MCP1, MIP1A, and TNFα than did those not requiring ICU admission, suggesting that the cytokine storm was associated with disease severity. However, 2019-nCoV infection also initiated increased secretion of T-helper-2 (Th2) cytokines (eg, IL4 and IL10) that suppress inflammation, which differs from SARS-CoV infection.22Wong CK Lam CWK Wu AKL et al.Plasma inflammatory cytokines and chemokines in severe acute respiratory syndrome. Further studies are necessary to characterise the Th1 and Th2 responses in 2019-nCoV infection and to elucidate the pathogenesis. Autopsy or biopsy studies would be the key to understand the disease.In view of the high amount of cytokines induced by SARS-CoV,22Wong CK Lam CWK Wu AKL et al.Plasma inflammatory cytokines and chemokines in severe acute respiratory syndrome., 24He L Ding Y Zhang Q et al.Expression of elevated levels of pro-inflammatory cytokines in SARS-CoV-infected ACE2+ cells in SARS patients: relation to the acute lung injury and pathogenesis of SARS. MERS-CoV,25Faure E Poissy J Goffard A et al.Distinct immune response in two MERS-CoV-infected patients: can we go from bench to bedside?., 26Falzarano D de Wit E Rasmussen AL et al.Treatment with interferon-α2b and ribavirin improves outcome in MERS-CoV-infected rhesus macaques. and 2019-nCoV infections, corticosteroids were used frequently for treatment of patients with severe illness, for possible benefit by reducing inflammatory-induced lung injury. However, current evidence in patients with SARS and MERS suggests that receiving corticosteroids did not have an effect on mortality, but rather delayed viral clearance.27Stockman LJ Bellamy R Garner P SARS: systematic review of treatment effects., 28Lansbury L Rodrigo C Leonardi-Bee J Nguyen-Van-Tam J Lim WS Corticosteroids as adjunctive therapy in the treatment of influenza., 29Arabi YM Mandourah Y Al-Hameed F et al.Corticosteroid therapy for critically ill patients with Middle East respiratory syndrome. Therefore, corticosteroids should not be routinely given systemically, according to WHO interim guidance.30WHOClinical management of severe acute respiratory infection when novel coronavirus (nCoV) infection is suspected. Among our cohort of 41 laboratory-confirmed patients with 2019-nCoV infection, corticosteroids were given to very few non-ICU cases, and low-to-moderate dose of corticosteroids were given to less than half of severely ill patients with ARDS. Further evidence is urgently needed to assess whether systematic corticosteroid treatment is beneficial or harmful for patients infected with 2019-nCoV.No antiviral treatment for coronavirus infection has been proven to be effective. In a historical control study,31Chu CM Role of lopinavir/ritonavir in the treatment of SARS: initial virological and clinical findings. the combination of lopinavir and ritonavir among SARS-CoV patients was associated with substantial clinical benefit (fewer adverse clinical outcomes). Arabi and colleagues initiated a placebo-controlled trial of interferon beta-1b, lopinavir, and ritonavir among patients with MERS infection in Saudi Arabia.32Arabi YM Alothman A Balkhy HH et al.Treatment of Middle East respiratory syndrome with a combination of lopinavir-ritonavir and interferon-β1b (MIRACLE trial): study protocol for a randomized controlled trial. Preclinical evidence showed the potent efficacy of remdesivir (a broad-spectrum antiviral nucleotide prodrug) to treat MERS-CoV and SARS-CoV infections.33Sheahan TP Sims AC Graham RL et al.Broad-spectrum antiviral GS-5734 inhibits both epidemic and zoonotic coronaviruses., 34Sheahan TP Sims AC Leist SR et al.Comparative therapeutic efficacy of remdesivir and combination lopinavir, ritonavir, and interferon beta against MERS-CoV. As 2019-nCoV is an emerging virus, an effective treatment has not been developed for disease resulting from this virus. Since the combination of lopinavir and ritonavir was already available in the designated hospital, a randomised controlled trial has been initiated quickly to assess the efficacy and safety of combined use of lopinavir and ritonavir in patients hospitalised with 2019-nCoV infection.Our study has some limitations. First, for most of the 41 patients, the diagnosis was confirmed with lower respiratory tract specimens and no paired nasopharyngeal swabs were obtained to investigate the difference in the viral RNA detection rate between upper and lower respiratory tract specimens. Serological detection was not done to look for 2019-nCoV antibody rises in 18 patients with undetectable viral RNA. Second, with the limited number of cases, it is difficult to assess host risk factors for disease severity and mortality with multivariable-adjusted methods. This is a modest-sized case series of patients admitted to hospital; collection of standardised data for a larger cohort would help to further define the clinical presentation, natural history, and risk factors. Further studies in outpatient, primary care, or community settings are needed to get a full picture of the spectrum of clinical severity. At the same time, finding of statistical tests and p values should be interpreted with caution, and non-significant p values do not necessarily rule out difference between ICU and non-ICU patients. Third, since the causative pathogen has just been identified, kinetics of viral load and antibody titres were not available. Finally, the potential exposure bias in our study might account for why no paediatric or adolescent patients were reported in this cohort. More effort should be made to answer these questions in future studies.Both SARS-CoV and MERS-CoV were believed to originate in bats, and these infections were transmitted directly to humans from market civets and dromedary camels, respectively.35Cui J Li F Shi Z-L Origin and evolution of pathogenic coronaviruses. Extensive research on SARS-CoV and MERS-CoV has driven the discovery of many SARS-like and MERS-like coronaviruses in bats. In 2013, Ge and colleagues36Ge X-Y Li J-L Yang X-L et al.Isolation and characterization of a bat SARS-like coronavirus that uses the ACE2 receptor. reported the whole genome sequence of a SARS-like coronavirus in bats with that ability to use human ACE2 as a receptor, thus having replication potentials in human cells.37Wang M Hu Z Bats as animal reservoirs for the SARS coronavirus: hypothesis proved after 10 years of virus hunting. 2019-nCoV still needs to be studied deeply in case it becomes a global health threat. Reliable quick pathogen tests and feasible differential diagnosis based on clinical description are crucial for clinicians in their first contact with suspected patients. Because of the pandemic potential of 2019-nCoV, careful surveillance is essential to monitor its future host adaption, viral evolution, infectivity, transmissibility, and pathogenicity.ContributorsBC and JW had the idea for and designed the study and had full access to all data in the study and take responsibility for the integrity of the data and the accuracy of the data analysis. YWa, GF, XG, JiXu, HL, and BC contributed to writing of the report. BC contributed to critical revision of the report. YWa, GF, XG, JiXu, and HL contributed to the statistical analysis. All authors contributed to data acquisition, data analysis, or data interpretation, and reviewed and approved the final version.Declaration of interestsAll authors declare no competing interests.Data sharingThe data that support the findings of this study are available from the corresponding author on reasonable request. Participant data without names and identifiers will be made available after approval from the corresponding author and National Health Commission. After publication of study findings, the data will be available for others to request. The research team will provide an email address for communication once the data are approved to be shared with others. The proposal with detailed description of study objectives and statistical analysis plan will be needed for evaluation of the reasonability to request for our data. The corresponding author and National Health Commission will make a decision based on these materials. Additional materials may also be required during the process.AcknowledgmentsThis work is funded by the Special Project for Emergency of the Ministry of Science and Technology (2020YFC0841300) Chinese Academy of Medical Sciences (CAMS) Innovation Fund for Medical Sciences (CIFMS 2018-I2M-1-003), a National Science Grant for Distinguished Young Scholars (81425001/H0104), the National Key Research and Development Program of China (2018YFC1200102), The Beijing Science and Technology Project (Z19110700660000), CAMS Innovation Fund for Medical Sciences (2016-I2M-1-014), and National Mega-projects for Infectious Diseases in China (2017ZX10103004 and 2018ZX10305409). We acknowledge all health-care workers involved in the diagnosis and treatment of patients in Wuhan; we thank the Chinese National Health Commission for coordinating data collection for patients with 2019-nCoV infection; we thank the International Severe Acute Respiratory and Emerging Infections Consortium (ISARIC) for sharing data collection templates publicly on the website; and we thank Prof Chen Wang and Prof George F Gao for guidance in study design and interpretation of results.Supplementary MaterialReferences1.Richman DD Whitley RJ Hayden FG Clinical virology.4th edn. ASM Press,\n Washington20162.Ksiazek TG Erdman D Goldsmith CS et al.A novel coronavirus associated with severe acute respiratory syndrome.N Engl J Med. 2003; 348: 1953-19663.Kuiken T Fouchier RAM Schutten M et al.Newly discovered coronavirus as the primary cause of severe acute respiratory syndrome.Lancet. 2003; 362: 263-2704.Drosten C Günther S Preiser W et al.Identification of a novel coronavirus in patients with severe acute respiratory syndrome.N Engl J Med. 2003; 348: 1967-19765.de Groot RJ Baker SC Baric RS et al.Middle East respiratory syndrome coronavirus (MERS-CoV): announcement of the Coronavirus Study Group.J Virol. 2013; 87: 7790-77926.Zaki AM van Boheemen S Bestebroer TM Osterhaus ADME Fouchier RAM Isolation of a novel coronavirus from a man with pneumonia in Saudi Arabia.N Engl J Med. 2012; 367: 1814-18207.WHOSummary of probable SARS cases with onset of illness from 1 November 2002 to 31 July 2003.8.WHOMiddle East respiratory syndrome coronavirus (MERS-CoV).9.WHONovel coronavirus – China.10.WHONovel coronavirus – Thailand (ex-China).11.WHONovel coronavirus – Japan (ex-China).12.WHONovel coronavirus – Republic of Korea (ex-China).13.CDCFirst travel-related case of 2019 novel coronavirus detected in United States.14.Tan W Zhao X Ma X et al.A novel coronavirus genome identified in a cluster of pneumonia cases — Wuhan, China 2019−2020.15.Sanz F Gimeno C Lloret T et al.Relationship between the presence of hypoxemia and the inflammatory response measured by C-reactive protein in bacteremic pneumococcal pneumonia.Eur Respir J. 2011; 38: 249216.Kidney disease: improving global outcomes (KDIGO) acute kidney injury work groupKDIGO clinical practice guideline for acute kidney injury.17.Garner JS Jarvis WR Emori TG Horan TC Hughes JM CDC definitions for nosocomial infections, 1988.Am J Infect Control. 1988; 16: 128-14018.Gao C Wang Y Gu X et al.Association between cardiac injury and mortality in hospitalized patients infected with avian influenza A (H7N9) virus.Crit Care Med. 2020; ()19.Perlman S Netland J Coronaviruses post-SARS: update on replication and pathogenesis.Nat Rev Microbiol. 2009; 7: 439-45020.Lee N Hui D Wu A et al.A major outbreak of severe acute respiratory syndrome in Hong Kong.N Engl J Med. 2003; 348: 1986-199421.Assiri A Al-Tawfiq JA Al-Rabeeah AA et al.Epidemiological, demographic, and clinical characteristics of 47 cases of Middle East respiratory syndrome coronavirus disease from Saudi Arabia: a descriptive study.Lancet Infect Dis. 2013; 13: 752-76122.Wong CK Lam CWK Wu AKL et al.Plasma inflammatory cytokines and chemokines in severe acute respiratory syndrome.Clin Exp Immunol. 2004; 136: 95-10323.Mahallawi WH Khabour OF Zhang Q Makhdoum HM Suliman BA MERS-CoV infection in humans is associated with a pro-inflammatory Th1 and Th17 cytokine profile.Cytokine. 2018; 104: 8-1324.He L Ding Y Zhang Q et al.Expression of elevated levels of pro-inflammatory cytokines in SARS-CoV-infected ACE2+ cells in SARS patients: relation to the acute lung injury and pathogenesis of SARS.J Pathol. 2006; 210: 288-29725.Faure E Poissy J Goffard A et al.Distinct immune response in two MERS-CoV-infected patients: can we go from bench to bedside?.PLoS One. 2014; 9e8871626.Falzarano D de Wit E Rasmussen AL et al.Treatment with interferon-α2b and ribavirin improves outcome in MERS-CoV-infected rhesus macaques.Nat Med. 2013; 19: 1313-131727.Stockman LJ Bellamy R Garner P SARS: systematic review of treatment effects.PLoS Med. 2006; 3: e34328.Lansbury L Rodrigo C Leonardi-Bee J Nguyen-Van-Tam J Lim WS Corticosteroids as adjunctive therapy in the treatment of influenza.Cochrane Database Syst Rev. 2019; 2: CD01040629.Arabi YM Mandourah Y Al-Hameed F et al.Corticosteroid therapy for critically ill patients with Middle East respiratory syndrome.Am J Respir Crit Care Med. 2018; 197: 757-76730.WHOClinical management of severe acute respiratory infection when novel coronavirus (nCoV) infection is suspected.31.Chu CM Role of lopinavir/ritonavir in the treatment of SARS: initial virological and clinical findings.Thorax. 2004; 59: 252-25632.Arabi YM Alothman A Balkhy HH et al.Treatment of Middle East respiratory syndrome with a combination of lopinavir-ritonavir and interferon-β1b (MIRACLE trial): study protocol for a randomized controlled trial.Trials. 2018; 19: 8133.Sheahan TP Sims AC Graham RL et al.Broad-spectrum antiviral GS-5734 inhibits both epidemic and zoonotic coronaviruses.Sci Transl Med. 2017; 9eaal365334.Sheahan TP Sims AC Leist SR et al.Comparative therapeutic efficacy of remdesivir and combination lopinavir, ritonavir, and interferon beta against MERS-CoV.Nat Commun. 2020; 11: 22235.Cui J Li F Shi Z-L Origin and evolution of pathogenic coronaviruses.Nat Rev Microbiol. 2019; 17: 181-19236.Ge X-Y Li J-L Yang X-L et al.Isolation and characterization of a bat SARS-like coronavirus that uses the ACE2 receptor.Nature. 2013; 503: 535-53837.Wang M Hu Z Bats as animal reservoirs for the SARS coronavirus: hypothesis proved after 10 years of virus hunting.Virol Sin. 2013; 28: 315-317Article InfoPublication HistoryPublished: January 24, 2020IdentificationDOI: https://doi.org/10.1016/S0140-6736(20)30183-5Copyright© 2020 Elsevier Ltd. All rights reserved.ScienceDirectAccess this article on ScienceDirectClinical features of patients infected with 2019 novel coronavirus in Wuhan, ChinaFiguresFigure 1Date of illness onset and age distribution of patients with laboratory-confirmed 2019-nCoV infectionFigure 2Timeline of 2019-nCoV cases after onset of illnessFigure 3Chest CT imagesTablesTable 1Demographics and baseline characteristics of patients infected with 2019-nCoVTable 2Laboratory findings of patients infected with 2019-nCoV on admission to hospitalTable 3Treatments and outcomes of patients infected with 2019-nCoV\n \n\n \n\n\n \n\n \n\n \n\n Linked Articles"],["topImageUrl",null],["error",null]]}]}],["text","轉po:\n\n預估兩週潛伏期\nLancet論文統計指出\n\n兩成不發燒\n七成會咳嗽\n症狀非常不典型！\n\n一週後發病\n發病後一週\n32% 加護病房\n29% ARDS插管急救\n15% 死亡終結\n\n意思是\n重症一半會死！\n\n機率大概\n比六發子彈\n直接開一槍的\n俄羅斯輪盤低一些～\n\n中國官方還告訴大家沒事兒不要怕？！\n\n我身為\n高雄榮總\n急診一線醫師\n加護病房主治醫師\n我目前想不到有什麼其他比這個更嚴重更糟糕的傳染病了🙏\n\n\nLancet\n\nhttps://www.thelancet.com/journals/lancet/article/PIIS0140-6736(20)30183-5/fulltext"],["id","1fbhbmuiiyn7x"],["createdAt","2020-01-27T10:59:00.386Z"]]}],["reply",{"__iterable":"Map","data":[["id","jmRj8W8Bd3n3h-WYx1yF"],["user",{"__iterable":"Map","data":[["id","AVriAlOoyrDaTqlmmqhC"],["name","Lin"],["avatarUrl",null],["level",22]]}],["type","RUMOR"],["text","15% 死亡終結並不正確，傳染病防治醫療網指揮官張上淳已提醒大家這樣的詮釋易造成誤解，因為 Lancet 的這份研究對象是針對疫情初期確診的41個「中度以上患者（皆已有肺炎）」，與臨床上仍有許多「不一定有肺炎的輕症患者」不同。"],["reference","https://pansci.asia/archives/178597?fbclid=IwAR1jdPHXfmZThAIsoGYr-MOtF-_92OQEMPwt_AUUCh60BVBpkfoQsA10PFc"],["hyperlinks",{"__iterable":"List","data":[{"__iterable":"Map","data":[["title",""],["url","https://pansci.asia/archives/178597?fbclid=IwAR1jdPHXfmZThAIsoGYr-MOtF-_92OQEMPwt_AUUCh60BVBpkfoQsA10PFc"],["summary",""],["topImageUrl",null],["error",null]]}]}],["createdAt","2020-01-29T13:00:45.833Z"]]}]]}]}],["searchArticles",{"__iterable":"List","data":[]}],["searchReplies",{"__iterable":"List","data":[]}]]}]]},"replyDetail":{"__iterable":"Map","data":[["state",{"__iterable":"Map","data":[["isLoading",false],["isAuthLoading",false],["isReplyLoading",false]]}],["data",{"__iterable":"Map","data":[["reply",null],["originalReplyConnection",null]]}]]},"auth":{"__iterable":"Map","data":[["state",{"__iterable":"Map","data":[["isLoading",false],["isDialogShown",false]]}],["user",null]]}}

Standard PLA+, Snow White, 1.75mm

現有回應

增加新回應

你可能也會對這些類似文章有興趣