2014年浙江省丽水市人感染H7N9禽流感病毒分离株基因特征分析

目的 测定浙江省丽水市1株H7N9禽流感病毒8个基因序列,从分子水平分析其基因特征和遗传变异特点。方法 对人感染H7N9禽流感确诊病例标本在P3实验室进行犬肾传代细胞培养;反转录-聚合酶链反应扩增病毒8个RNA片段后测定基因序列;通过生物信息学软件将8个序列与GISAID和GenBank数据库下载的序列进行多重比对和匹配,发现基因特征,分析病毒关键位点的分子变异情况,最大似然法绘制系统进化树用于基因进化分析。结果 从病例标本中分离到病毒:A/Zhejiang/LS01/2014(H7N9),测序获得8个基因核苷酸全序列。HA裂解位点只包含2个碱性氨基酸,HA蛋白受体结合关键氨基酸位点发生A134T、G186V和Q226L变异;在PB2蛋白上发现E627K突变;M2蛋白上发现耐药位点S31N。LS01株HA基因与A/chicken/Zhejiang/DTID-ZJU01/2013(H7N9)的同源性最高(98.90%),NA基因与A/chicken/Changzhou/C08/2013(H9N9)的同源性最高(98.60%),LS01株其他6个编码内部蛋白的基因均与禽源H9N2株高度同源(99.10%~99.89%)。LS01株HA和NA基因与沪苏浙一带流行的H7N9禽流感病毒株位于同一亚分支,有着直接的进化关系,在相近地域有进化关系较近的禽类流感病毒存在。结论 A/Zhejiang/LS01/2014(H7N9)株8个基因序列均符合禽源H7N9亚型禽流感病毒特征,其病原基因均为禽类来源。HA裂解位点具有低致病性禽流感病毒基因特点,Q226L和E627K位点发生变异可能与其对人具有高致病力相关。     

新型甲型H7N9禽流感病毒HA与NA的进化分析

摘要：目的：研究2013年甲型H7N9流感病毒血凝素（HA）与神经氨酸酶（NA）的基因进化特征,探讨该病毒的遗传变异和分子特性。 方法：收集GenBank中已登录的禽流感病毒基因相关参考序列,利用生物信息学软件DNAMAN和MEGA 4.0对HA和NA基因序列进行进化分析,并利用Phymol 软件进行同源建模。 结果：2013年甲型H7N9流感病毒HA与NA的氨基酸序列与国际/国内参考毒株的同源性为95.7%~99.8%。同源建模发现该毒株HA的氨基酸序列发生了Q226L突变,NA上也发生了R294K和69 5N del氨基酸突变。 结论：新型甲型H7N9流感病毒的HA和NA基因序列与我国江浙地区和部分东亚国家的H7N9禽流感病毒参考株具有高度的同源性,该毒株HA与NA蛋白氨基酸序列发生的变异,可能是病毒致病性改变的原因之一。     

2016-2017年我国甲型流感病毒H3N2亚型HA和NA基因的分子特征分析

目的探讨2016-2017年我国甲型流感病毒H3N2亚型血凝素(hemagglutinin,HA)和神经氨酸酶(neuraminidase,NA)基因的分子特征,为甲型流感病毒H3N2亚型的防控提供科学依据。方法从全球共享禽流感数据库(GISAID)中获得我国2016~2017年分离的525株甲型流感病毒H3N2亚型病毒和13株H3N2亚型参考株的HA和NA基因序列。应用MEGA 6.0软件分析HA和NA的分子进化特征、氨基酸位点和耐药位点的变异情况。结果进化树分析显示13株参考株之间HA和NA氨基酸的同源性均为96.9%~99.1%。2016-2017年我国甲型流感病毒H3N2亚型的病毒株HA和NA氨基酸序列之间同源性均为96.3%~100%,HA和NA的优势亚分支均为3C.2a.1,相当一部分毒株分型不够明确。HA氨基酸变异分析显示9株发生N121E突变,103株发生N121K的突变且主要来源于2017年的香港毒株;182株发生G/R142K突变;A/Hong_Kong/3079/2017-HA和A/Guangdong-Chengguan/1383/2017-HA均发生A128I突变;NA蛋白仅3株(A/Hunan-Suxian/1980/2016-NA、A/Hunan-Yuhua/11799/2016-NA和A/Ningxia-Yuanzhou/1657/2016-NA)发现H275Y耐药位点突变。结论 2016-2017年我国甲型流感病毒H3N2亚型的HA和NA蛋白与参考株间存在一定的差异,优势亚型均为3C.2a.1;与病毒的毒力及药物相关的部分关键氨基酸位点发生变异。应密切关注H3N2病毒的分子特征,为其防控提供参考。     

2013～2016年东部沿海地区甲型H1N1流感病毒分子特征分析

摘要：目的：通过分析东部沿海地区（江苏、浙江和上海）甲型H1N1流感病毒［A(H1N1) pdm09］的分子特征,为防控A(H1N1) pdm09流感提供科学依据。 方法：从GISAID数据库中获得江浙沪地区2013～2016年分离的A(H1N1) pdm09病毒株和WHO推荐疫苗株的血凝素（HA）和神经氨酸酶（NA）基因序列。运用MEGA6.0软件分析HA和NA的分子特征、关键氨基酸位点和耐药位点的变异情况。 结果：进化树分析显示江浙沪地区A(H1N1) pdm09病毒株与WHO参考株的HA、NA基因序列差异较小,与WHO推荐疫苗株A/California/07/2009(H1N1)间的同源性分别为97.0%～99.5%和97.9%～99.6%;与WHO推荐疫苗株A/South_Africa/3626/2013(H1N1)间的同源性分别为98.0%～99.8%和98.6%～99.9%。病毒氨基酸变异分析显示70株A(H1N1)pdm09病毒的HA蛋白均发生83位(P83S)、203位(S203T)和321位(I321V)突变,且2016年分离株出现A195V突变;HA的裂解位点未发生变异,仍为IQSR↓GLF;2013年有2株NA蛋白出现H275Y耐药位点的变异。 结论：2013～2016年东部沿海地区A(H1N1) pdm09流感病毒的HA和NA蛋白与WHO推荐疫苗株具有高度同源性,关键的氨基酸位点发生变异与病毒的毒力及对药物的敏感性相关,因此我们要密切关注A(H1N1) pdm09病毒,防止其再次发生暴发。     

甘肃省人感染H7N9禽流感病毒基因组对比分析

  目的  分析甘肃省发现的人感染H7N9禽流感病毒序列,对比高致病性禽流感（HPAI）和低致病性禽流感（LPAI）H7N9病毒的基因变异及分子进化特征,为疾病预防控制提供参考。  方法  对甘肃省2017年以来发现的人感染H7N9禽流感病毒进行基因组对比,并使用MEGA 7.0等软件分析其全基因组特征。  结果  6例人感染H7N9病例,均有活禽或其环境暴露史,共获得3株毒株:GS/19545、GS/27151和GS/23275。 GS/19545和GS/27151为LPAI,GS/23275为HPAI。 3株毒株的HA、NA基因均来自欧亚系的长三角支系。 GS/23275的NP基因和广东省2017年分离的HPAI处于同一进化分支上,与GS/19545、GS/27151形成两个明显的分支。 GS/23275的PB2基因与安徽省2015年从禽类体内分离的H9N2病毒亲缘关系较近。 3株毒株的HA受体结合位点均发生G186V、S138A突变,NA蛋白茎部均发生68～72位点氨基酸缺失,在PA、PB1、PB2、M1、M2、NS1、NS2等蛋白中均发生相同氨基酸突变。  结论  甘肃省人感染H7N9禽流感病毒基因组发生了部分重要分子突变,可能导致其毒力增强,并具备潜在的人际传播能力。 加强禽流感病毒基因特征研究分析,有助于从分子水平上监测病毒变异突变并对疫情进行科学防控。     

人感染高致病性H5N6禽流感病毒的分子生物学特征

目的分析2014-2015年3例新型人感染高致病性禽流感(HPAI)H5N6毒株的血凝素(HA)与神经氨酸酶(NA)基因序列,探讨该病毒的遗传变异及分子生物学特征。方法收集Gen Bank、全球共享禽流感数据倡议组织(GISAID)及浙江大学传染病诊治国家重点实验室提供的H5N6病毒基因序列,利用生物信息软件MEGA6.0及Net Nglyc服务器对基因序列进行遗传进化和分子变异特征分析。结果 3例感染HPAI-H5N6患者毒株的HA与NA氨基酸序列的同源性分别为96.5%~98.7%和90.1%~98.2%;HA氨基酸序列比对分析发现A/Guangzhou/39715/2014(H5N6)和A/Sichuan/26221/2014(H5N6)与A/duck/Eastern China/1111/2011(H5N2)同源性最近,分别为97.7%和98.5%;A/Yunnan/0127/2015(H5N6)与A/muscovy duck/Vietnam/LBM631/2014(H5N1)同源性最近(97.2%);NA氨基酸序列比对分析发现A/Guangzhou/39715/2014(H5N6)和A/Yunnan/0127/2015(H5N6)与A/swine/Guangdong/K6/2010(H6N6)同源性最近,分别为97.2%和97.6%,A/Sichuan/26221/2014(H5N6)与A/duck/Guangdong/S3073/2010(H6N6)同源性最近(97.4%);进化树分析显示3例患者毒株处于2个不同的分支;HA基因包含有S137A、T160A等突变,糖基化分析显示有6~7个N-糖基化位点。结论新型人感染HPAI-H5N6病毒是一种重组病毒,其HA基因来源于亚洲禽类H5亚型,NA基因来源于亚洲禽类H6N6;HA基因位点突变及N-糖基化改变可能有助于增强对人类的感染。     

2017年北京市大兴区首例人感染H7N9禽流感病例感染来源分析

目的 对2017年北京市大兴区首例人感染H7N9禽流感确诊病例的感染来源及部分基因序列进行分析,为人感染H7N9禽流感疫情防控提供依据。 方法 采用现场流行病学调查和实验室检测相结合的方法,追溯病例的感染来源。 结果 病例发病前1周有活禽接触史,病例咽拭子、深咳痰液、血液标本人感染H7N9禽流感核酸均为阳性,在暴露的活禽及外环境中检出H7N9禽流感病毒核酸阳性,7名共同暴露者及密切接触者核酸检测为阴性。 血凝素（HA）基因和神经氨酸酶（NA）基因测序分析显示未发生明显变异。 结论 活禽暴露是该病例感染的关键风险因素,病例的感染来源是购自某流动集市的活禽,病毒基因测序及HA0裂解位点测定,该病毒属于禽H7N9长江三角洲分支病毒,为低致病性禽流感病毒。      

2014－2017年广州市禽类市场外环境H7N9禽流感病毒监测与遗传进化分析

目的 对广州市禽类市场外环境H7N9禽流感病毒流行情况和基因进化特点进行监测分析,为人感染H7N9禽流感防控提供研究数据。 方法 采集2014 — 2017年广州市禽类市场外环境标本,采用实时荧光定量RT-PCR方法检测H7N9禽流感病毒并绘制血凝素（HA）基因遗传进化树,分析H7N9禽流感病毒流行特点和基因进化特点。 结果 2014 — 2017年累计检测外环境标本28 252份,检出H7N9阳性标本888份,阳性率为3.14%。 检出率2015年最低,2014年最高,分别为1.60%和3.89%。2 — 5月出现H7N9禽流感病毒流行高峰,4月检出率最高为13.00%。 测序分析结果显示,HA基因核苷酸同源性为88.6% ~ 100%,氨基酸同源性为92.0% ~ 100%。 裂解位点分析发现,2017年外环境开始出现高致病性H7N9病毒。 进化分析发现,HA基因呈现多进化分支特点。 结论 2014 — 2017年广州市禽类市场外环境H7N9禽流感病毒持续存在且病毒变异较快,呈现基因多样性。 2017年外环境已发现高致病性H7N9病毒,近年病毒HA基因归属于华南分支,呈现本土化,同时监测到来源于野鸟的另一独立分支的病毒存在,提示加强本地区野鸟H7N9禽流感病毒监测的必要性。      

广东人类禽流感H5N1亚型血凝素和神经氨酸酶基因同源性分析

目的采用生物信息学技术,寻找广东地区人类禽流感H5N1亚型基因同源毒株。方法检测广东人禽流感H5N1亚型HA和NA基因核苷酸序列,从GenBank下载国内各种动物和人类相应基因序列,采用Mega5.03和Lasergene7.1软件进行核苷酸同源性和氨基酸变异分析。结果在总共174个HA基因和173个NA基因序列中,发现广东毒株A/Guangdong/01/2006的HA和NA基因与湖南鸡和安徽鸭流感毒株同源性较高;广东毒株A/Guangdong/02/2006的HA和NA基因与香港鸟禽类流感毒株同源性较高。同源性较高的毒株基因之间,氨基酸位点存在差异,但尚未显示流行病学和临床意义。结论广东A/Guangdong/01/2006毒株与湖南鸡和安徽鸭流感毒株具有共同的最近祖先,广东A/Guangdong/02/2006毒株与香港鸟禽类流感毒株具有共同的最近祖先。     

2009~2012年杭州地区甲型H1N1流感病毒HA与NA基因的进化分析

摘要：目的：研究杭州地区2009年至2012年甲型H1N1流感病毒血凝素（HA）与神经氨酸酶（NA）的基因进化特征,分析该病毒的遗传变异和抗原的分子水平转变。 方法：用RT-PCR分别扩增甲型H1N1流感病毒HA基因和NA基因片段并进行测序,用DNAMAN和MEGA 4.0生物信息学软件对HA和NA基因序列进行进化分析。 结果：建立的RT-PCR可成功检测HA(C)、HA(D)和NA基因,遗传进化分析结果表明,杭州地区甲型H1N1流感病毒的HA与NA氨基酸序列的亲缘系数与WHO推荐的病毒株及国内代表株的同源性均为98.9%~100%;三维构象结果表明,HA编码的蛋白质有5个位点发生改变,而NA编码的蛋白质仅发生2个位点改变。 结论：2009年至2011年杭州地区甲型H1N1流感病毒HA和NA基因序列与世界范围内流行的病毒参考株具有高度同源性,但HA与NA蛋白质表位存在某些氨基酸位点的改变。     

Guns and Knives in New Mexico: Patterns of Penetrating Trauma, 1978–1993

Objective: To identify patterns of nonfatal and fatal penetrating trauma among children and adults in New Mexico using ED and medical examiner data.
Methods: The authors retrospectively sampled in 5-year intervals all victims of penetrating trauma who presented to either the state Level-1 trauma center or the state medical examiner from a 16-year period (1978–1993). Rates of nonfatal and fatal firearm and stabbing injury were compared for children and adults.
Results: Rates of nonfatal injury were similar (firearm, 34.3 per 100,000 person-years; stabbing, 35.1). However, rates of fatal injury were significantly different (firearm, 21.9; stabbing, 2.7; relative risk: 8.2; 95% confidence interval: 5.4, 12.5). From 1978 to 1993, nonfatal injury rates increased for children (p = 0.0043) and adults (p < 0.0001), while fatal penetrating injury remained constant. The increase in nonfatal injury in children resulted from increased firearm injury rates. In adults, both stabbing and firearm nonfatal injury rates increased.
Conclusions: Nonfatal injury data suggest that nonfatal violence has increased; fatal injury data suggest that violent death rates have remained constant. Injury patterns vary by age, mechanism of trauma, and data source. These results suggest that ED and medical examiner data differ and that both are needed to guide injury prevention programs.     

Interprofessional approaches to creating safe,high quality health care

Three supplementary perspectives are presented arguing that interprofessional collaboration is both necessary and desirable. Nonetheless, there are often too many serious intra-professional barriers and obstacles to interprofessional collaboration to make it successful. Some of these barriers, it is argued and illustrated, are found in the multiple ways in which professional identity is tacitly acquired and embodied in the practitioners' habitual, everyday practice. The paper then explores ways in which reflection, especially Second order reflection, can help to elucidate and overcome these obstacles, as well as increasing professional adaptability and competence.     

Systematic Reviews Published in the January 2011 Issue of The Cochrane Library

ABSTRACT

The Cochrane Library of Systematic Reviews is published quarterly as a DVD and monthly online. The January 2011 issue (first quarterly DVD for 2011) contains 4515 complete reviews, 1985 protocols for reviews in production, and 13,521 one-page summaries of systematic reviews published in the general medical literature. In addition, there are citations of 641,000 randomized controlled trials, and 14,018 cited papers in the Cochrane methodology register. The health technology assessment database contains over 9300 citations. One hundred and seven new reviews have been published in the last 3 months, of which five have potential relevance for practitioners in pain and palliative medicine.     

Determination of creatine kinase isoenzyme MB activity in serum using immunological inhibition of creatine kinase M subunit activity. Activity kinetics and diagnostic significance in myocardial infarction.

This is a new method for the determination of creatine kinase isoenzyme MB activity in serum. The method uses direct activity measurement of creatine kinase B subunit activity after blocking of CK-M subunit activity by inhibiting antibodies. The test takes no longer than 15 min. The method yields an intra-serial C.V. of 2.0-12.9%, and a C.V. from day to day of 5.5%. The detection limit is 3.4 U/l creatine kinase MB. In the 95 cases with proven myocardial infarction several types of creatine kinase MB activity kinetics could be determined. The percentage of creatine kinase MB of peak CK-total is 6-25%, with a mean of 11.1%. The amount of creatine kinase MB with respect to total CK activity after reinfarction is higher than the amount after initial infarction.     

俯卧位通气对高海拔地区肺复张治疗无效急性呼吸窘迫综合征患者氧合的影响

目的 探讨俯卧位通气对高海拔地区肺复张术(RM)治疗无效急性呼吸窘迫综合征(ARDS)患者的治疗作用.方法 从海拔2260m的地区医院筛选RM治疗无效的41例ARDS患者[平均氧合指数( PaO2/FiO2)较RM前升高＜20％视为RM无效],依不同病因分为肺内源性ARDS组(ARDSp组)和肺外源性ARDS组(ARDSexp组),每组再按信封法随机分为俯卧位组和仰卧位组,即ARDSp俯卧位组(11例)、ARDSp仰卧位组(9例)、ARDSexp俯卧位组(10例)、ARDSexp仰卧位组(11例).在通气前及通气1、2、3、4h监测动脉血氧分压( PaO2)、PaO2/FiO2、静态顺应性(Cst)、气道阻力(Raw)的变化.结果 通气lh时,ARDSexp俯卧位组PaO2/FiO2( mm Hg,l mm Hg=0.133 kPa)即较通气前显著升高(157.4±40.6比129.3±48.7,P＜0.05),并随通气时间延长呈持续增高趋势,4h达峰值(219.1 ±41.1);且ARDSexp俯卧位组通气3h内PaO2/FiO2较其他3组显著增高,另3组间则差异无统计学意义.ARDSp俯卧位组、ARDSexp俯卧位组通气4h时PaO2/FiO2均较相应仰卧位组显著增高(208.8±39.7比127.4±47.1,219.1±41.1比124.9±50.8,均P＜0.05).4组通气前后Cst无显著改变,各组间差异也无统计学意义.ARDSp俯卧位组通气4h时Raw(cmH2O·L-1·s-1)较通气前显著降低(6.8±1.7比10.7±1.8,P＜0.05),且明显低于其他3组;其他3组各时间点Raw组内及组间比较差异均无统计学意义.结论 俯卧位通气作为ARDS机械通气重要策略之一,可以改善RM无效高原ARDS患者的氧合,为抢救患者赢得宝贵的时间.     

Digital imaging among medical facilities

The Department of Veterans Affairs (VA) in the USA operates a network of 172 medical centres which all utilize a hospital information system (HIS) which has been developed and is currently maintained by the VA. During the past several years, an image management and communication module has been developed, installed and clinically utilized at the Washington DC and Maryland VA Medical Centres. This image management and communication system, referred to as the decentralized hospital computer program (DHCP) imaging system, is fully integrated with a commercial picture archiving and communication system (PACS). The system is utilized to capture, archive, and display all images generated within the hospital including radiology, nuclear medicine, pathology, endoscopy, bronchoscopy, and dermatology, intraoperative photographs, ECG data, and a limited number of paper documents. The ultimate goal of the project is to have all patient text and image data available at any clinical workstation to any authorized user anywhere within the network of medical centres. Clinical requirements for an imaging workstation include ease of use, rapid and reliable access to the complete set of patient information, and images which are of acceptable quality to meet the requirements of the user and the subspecialty. Patient confidentiality and data security must be safeguarded at all times. Integration of the images with the remainder of the patient's database was found to be critical to the success of the project. The experience at the Washington and Maryland facilities suggests that an imaging system that is successfully integrated with a hospital information system can provide substantial clinical and economic benefits both within and among medical centres. Clinical acceptance and utilization of the system has been excellent, particularly in diagnostic radiology where DHCP Imaging has been interfaced to a commercial PAC system. Based upon this initial experience, the VA has begun to deploy the system throughout its large network of medical centres.     

Myocardial elastography at both high temporal and spatial resolution for the detection of infarcts

Myocardial elastography is a novel method for noninvasively assessing regional myocardial function, with the advantages of high spatial and temporal resolution and high signal-to-noise ratio (SNR). In this paper, in-vivo experiments were performed in anesthetized normal and infarcted mice (one day after left anterior descending coronary artery [LAD] ligation) using a high-resolution (30 MHz) ultrasound system (Vevo 770, VisualSonics Inc., Toronto, ON, Canada). Radiofrequency (RF) signals of the left ventricle (LV) in longitudinal (long-axis) view and the associated electrocardiogram (ECG) were simultaneously acquired. Using a retrospective ECG gating technique, 2-D full field-of-view RF frames were acquired at an extremely high frame rate (8 kHz) that resulted in high-quality incremental displacement and strain estimation of the myocardium. The incremental results were further accumulated to obtain the cumulative displacements and strains. Two-dimensional and M-mode displacement images and strain images (elastograms), as well as displacement and strain profiles as a function of time, were compared between normal and infarcted mice. Incremental results clearly depicted cardiac events including LV contraction, LV relaxation and isovolumetric phases in both normal and infarcted mice, and also evidently indicated reduced motion and deformation in the infarcted myocardium. The elastograms indicated that the infarcted regions underwent thinning during systole rather than thickening, as in the normal case. The cumulative elastograms were found to have higher elastographic SNR (SNR(e)) than the incremental elastograms (e.g., 10.6 vs. 4.7 in a normal myocardium, and 6.0 vs. 2.4 in an infarcted myocardium). Finally, preliminary statistical results from nine normal (m = 9) and seven infarcted (n = 7) mice indicated the capability of the cumulative strain in differentiating infracted from normal myocardia. In conclusion, myocardial elastography could provide regional strain information at simultaneously high temporal (>/=0.125 ms) and spatial ( approximately 55 microm) resolution as well as high precision ( approximately 0.05 microm displacement). This technique was thus capable of accurately characterizing normal myocardial function throughout an entire cardiac cycle, at the same high resolution, and detecting and localizing myocardial infarction in vivo.     

Prise en charge des entorses externes de cheville : étude clinique préliminaire sur l’efficacité du dispositif Myolux™

Ankle sprains are the most common injury of the musculoskeletal system and are associated with significant societal and economic impacts. It has been proven that classical therapeutic strategies may not be effective in preventing recurrent injuries: the recurrence rates reported in the literature can reach 73%. In order to provide an effective rehabilitation solution, a destabilizing orthosis was developed. This device is equipped with a mechanical articulator reproducing the subtalar mechanics and placed under the heel. In this paper, we present the main results of a preliminary clinical study conducted between 2004 and 2007. All subjects included in this study were treated with the abovementioned orthosis during 10 rehabilitation sessions of 30 minutes each. Data show a relatively low recurrence rate of 12% for the overall population. Moreover, it's of primary importance to note that this satisfactory ratio is largely reduced (3% of recurrence rate) for the 29 patients who performed one training session per month after the 10th initial rehabilitation sessions. Hence, the destabilizing orthosis appears to be an effective solution to prevent recurrent ankle sprains. However, joint protection requires long-term and regular training sessions. This result has motivated the development of a similar device allowing patients to perform training sessions at home. Finally, data obtained in this study are promising awaiting the final results of the comparative, multicentric and independent clinical trials currently managed by the Hospices Civils de Lyon.