盖塔病毒分子进化及其迁徙

目的了解世界各地分离盖塔病毒分子进化特征及其时空迁徙。方法用Clustal X1.83、MegaAlign和Gene DOC软件对测定的核苷酸和推测的氨基酸序列进行比较分析,用MEGA 6.0软件绘制系统发生树,采用BEAST v 1.8.1软件包里的Bayesian Stochastic Search Variable Selection(BSSVS)程序进行盖塔病毒的空间动力学分析。结果盖塔病毒E2基因全长均为1 266nt,编码422aa,其核苷酸和氨基酸同源性分别为94.5%~100%和96.4%~100%。病毒分子进化分析发现,盖塔病毒不存在蚊虫,马匹和猪等宿主动物和媒介的种属和地域分布差异。生物信息学分析结果显示,盖塔病毒起源于马来西亚,此后依次传播至日本、中国、韩国以及蒙古国和俄罗斯等地。结论自1955年首次分离盖塔病毒至2014年间分离的盖塔病毒E2基因序列较为稳定,未发现病毒基因组之间存在种属及地域分布差异,并且该病毒已经从热带地区传播到欧亚大陆多个国家和地区,因此加强盖塔病毒及人畜动物感染的检测和监测至关重要。     

四川省猪源盖塔病毒的分离鉴定及遗传进化分析

目的 对检测到的疑似盖塔病毒进行分离鉴定和全基因序列遗传进化分析。方法 从四川某猪场1例混合感染猪繁殖与呼吸综合征的仔猪血液中,进行RT-RCR检测,检测到GETV阳性。对样品进行无菌处理后,接种至BHK21细胞,连续传代培养,通过细胞病变、蚀斑纯化、RT-RCR扩增和全基因序列测定鉴定细胞培养物。结果 我们成功分离到四川首株猪盖塔病毒,并将其命名为SC201807;对该病毒全基因扩增测序并进行序列分析,发现SC201807与GenBank中上传的35株GETV序列相似性为99.0%~96.1%,E2序列高度保守仅在E310(天冬氨酸→谷氨酸)发生突变。全基因组核苷酸以及E2氨基酸序列遗传进化分析显示SC201807株与中国猪源HNJZ-S2及日本蚊源15-I-752、15-I-1105、14-I-605-C2、14-I-605-C1、12IH26亲缘关系最近,处于同一进化分支。结论 SC201807株扩充了猪源GETV的感染谱,对其致病性、感染和传播机制、环境微生物学及公共卫生等相关研究提供素材。     

甘肃省天水及陇南部分地区虫媒病毒调查

目的对甘肃省天水陇南部分地区的吸血蚊虫进行病毒分离与鉴定。方法2006年8月在当地采集蚊虫标本,分离病毒和对病毒分离物进行血清学和分子生物学鉴定,以软件进行病毒的核苷酸序列比对和系统发生分析。结果分离到19株病毒,鉴定结果显示分离株GS10-2为盖塔病毒,GS42-2为版纳病毒,其余分离株为一种基因组约8 000核苷酸的未知RNA病毒。盖塔病毒分离株3’UTR中具有与已往中国分离株相同的缺失序列和3个特异核苷酸位点。版纳病毒分离株基因组第12片段的进化关系同其它中国分离株有明显差异,位于一条独立的进化枝中。结论在该地区分离到1株盖塔病毒、1株版纳病毒和17株未知RNA病毒;盖塔病毒与以往我国分离株的遗传关系密切,版纳病毒与我国其它地区分离株有明显差异,在进化上相对独立。     

盖塔病毒研究状况

盖塔病毒属于披膜病毒科甲病毒属的一种人畜共患虫媒病毒,主要分布于日本,东南亚及澳大利亚等地。该病毒主要引起马和猪的感染。目前,我国从海南、云南、甘肃及河北等地均分离到该病毒,但对该病毒在我国的流行特点及其与人畜疾病的关系有待进一步研究。本文对盖塔病毒病分子生物学特性﹑分布﹑媒介及其临床特点等方面研究进展作出综述。     

血清hsCRP水平与糖尿病冠状动脉弥漫病变关系探讨

目的:研究炎症因子高敏C反应蛋白(hsCRP)、白介素-6(IL-6)和肿瘤坏死因子(TNF)-α与糖尿病冠状动脉弥漫病变关系。方法:根据冠状动脉造影结果及有无糖尿病共296例患者分成五组:单纯冠心病冠脉局灶病变组(I组)92例,单纯冠心病冠脉弥漫病变组(II组)78例,单纯糖尿病组(III组)30例,2型糖尿病合并冠状动脉局灶病变组(IV组)32例,2型糖尿病合并冠状动脉弥漫病变组(V组)64例。检测与比较各组患者血清hsCRP、IL-6、TNF-α的水平。结果:V组hsCRP水平明显高于I、II、III、IV组(14.65±8.74vs.7.78±5.91,9.16±8.13,7.47±4.62,10.58±7.43mg/L),均P0.01;IV组hsCRP水平高于I组和III组,P0.05。IL-6水平V组高于II、III组,均P0.05;TNF-α水平IV组(77.7±41.65)高于II、III组,均P0.05;IV组IL-6水平和V组TNF-α水也高于I、II、III组,但无统计学意义;在I、II组间三者均无明显差异。hsCRP与弥漫病变和糖尿病呈正相关。结论:糖尿病患者hsCRP水平的升高不仅是糖尿病血管并发症发生的预测因子也是糖尿病冠状动脉弥漫病变的预测因素。     

The effect of interleukin-6 on bacterial translocation in acute canine pancreatitis

Summary Background. Bacterial translocation from the gut to mesenteric lymph nodes and other extraintestinal sites is an important source of infection in acute pancreatitis. Impaired host immunity is known to promote bacterial translocation. Interleukin-6 (IL-6) is a multifunctional cytokine that regulates the immune response, acute phase reaction, and hematopoiesis. Methods. Twenty-four mongrel dogs (18–29 kg) were studied in four equal groups. In Groups I and II, acute pancreatitis was induced by direct pressure injection of 4% taurocholate and trypsin into the pancreatic duct at laparotomy. Groups III and IV had only laparotomy. Group I and III dogs were given IL-6 (50 μg/kg/d, sq) daily starting 24 h after operation and Group II and IV dogs received an equal volume of saline administered at similar time. All animals had blood drawn for culture, complete blood count (CBC), platelets, erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), and amylase on d 0, 1, 4, and 7. On d 7, mesenteric lymph nodes (MLN), spleen, liver, pancreas, and cecum were harvested for pathology study and for cultures of aerobic and anaerobic bacteria. Quantitative cecal cultures of aerobic and anaerobic bacteria were obtained. Results. All Group I and Group II dogs had severe pancreatitis. The increase of plasma CRP in Group I was sustained throughout treatment (1.3±0.3 on d 0 vs 3.1±0.3*, 3.0±0.3*, and 2.9±0.3* and d 1,4, and 7, respectively). Plasma CRP was increased in Group II on d 1 and d 4 (1.3±0.3 mg/dL on d 0 vs 3.6±0.3* mg/dL on d 1, and 3.1±0.3* on d 4, *p<0.05). There were no differences in white blood cell (WBC) count, differential, platelets, and ESR between Groups I and II. Bacterial translocation to MLN was lower in Group I (1/6) than in Group II (6/6) (p<0.05). All 6 dogs in Group II had bacterial spread to distant sites compared to 2 of 6 dogs in Group I (p=0.066). Both MLN and other distant organ cultures were negative in Group III and only 1 of 6 MLN cultures was positive in Group IV. Conclusions. IL-6 treatment decreases bacterial translocation to MLN and may be beneficial in reducing septic complications in acute pancreatitis.     

病毒性脑炎在马来西亚与新加坡

近期病毒性脑炎在马来西亚持续暴发流行,目前上报到马来西亚卫生部的患者总数已达234人,其中90人死亡。在这次流行中除有日本乙型脑炎病毒可检出外,主要检出了一种新的副粘病毒,这种新病毒被称谓类Hendra病毒,它与1994年在澳大利亚首次分离到的马麻疹病毒十分相象;类Hendra病毒在人和猪中感染,并在人群中引起脑炎,患此病致死者多与惊厥,肺部充血、水肿和脑水肿、脑疝有     

首次从云南蚊虫分离到辛德毕斯病毒及其鉴定

目的了解云南省虫媒病毒的存在和流行情况。方法2005年8月,在云南省勐海县采集蚊虫,蚊虫标本经研磨后,上清液接种BHK21细胞以分离病毒,用间接免疫荧光试验和RT-PCR等方法进行鉴定,并对病毒的分子生物学特征进行分析。结果采集到蚊虫标本9400只,其中分离到一株对BHK-21细胞能产生明显细胞病变的病毒(MX10)。该病毒经间接免疫荧光试验提示为甲病毒。用甲病毒属特异引物和Sindbis病毒E1基因特异引物对MX10病毒的RT-PCR扩增为阳性,经核酸序列测定分析证实该序列与Sindbis病毒泰国分离株(AF492770)同源性最高,为90.0%;与1987年分离自云南发热患者的YN87448株和1991年新疆按蚊分离株XJ-160的同源性分别为73.1%和72.0%。结论本次从勐海县蚊虫分离到的MX10病毒株为Sindbis病毒,并可能是Sindbis病毒的新亚型或新株系。     

甘肃省庆阳市媒介蚊虫类携带病毒调查

目的了解甘肃省庆阳市区域媒介蚊虫类分布特点及携带病毒情况,为防控虫媒病毒相关疾病提供科学依据。方法采用电子诱蚊灯、捕虫网诱捕,并进行分类;用细胞培养法分离病毒,用血清学分子生物学方法进行病毒鉴定。结果2012年捕获蚊虫5种,4238只,其中淡色库蚊2459只,占58．0％;中华按蚊794只,占18．73％;三带喙库蚊652只,占15．38％;刺扰伊蚊215只,占5．08％;济南按蚊118只,占2．8％;捕获的三带喙库蚊中分离出1株盖塔病毒,淡色库蚊检出6株库蚊黄病毒。结论淡色库蚊是庆阳市区域的优势蚊种,蚊虫携带黄病毒和盖塔病毒等虫媒病毒,应加大对该地区蚊虫及虫媒病毒的监测、调查、研究及防治工作。     

2004年孟加拉尼帕病毒病疫情简介

尼帕病毒(Nipah virus)是一种新发现的副粘病毒。在生物安全上属于最危险的第四级(BSL4)病毒。1998年至1999年尼帕病毒在马来西亚的猪群流行，并导致265人感染，其中105人死亡，引起世界各国广泛关注。尼帕病毒病是我国重点防范的一种人畜共患疫病。     

Genomic and In Vitro Phenotypic Comparisons of Epidemic and Non-Epidemic Getah Virus Strains

Getah virus is an emerging mosquito-borne animal pathogen. Four phylogenetic groups of GETV, Group I (GI), GII, GIII and GIV, were identified. However, only the GETV GIII was associated with disease epidemics suggesting possible virulence difference in this virus group. Here, we compared the genetic and in vitro phenotypic characteristics between the epidemic and non-epidemic GETV. Our complete coding genome sequence analyses revealed several amino acid substitutions unique to the GETV GIII and GIV groups, which were found mainly in the hypervariable domain of nsP3 and E2 proteins. Replication kinetics of the epidemic (GIII MI-110 and GIII 14-I-605) and non-epidemic GETV strains (prototype GI MM2021 and GIV B254) were compared in mammalian Vero cells and mosquito C6/36 and U4.4 cells. In all cells used, both epidemic GETV GIII MI-110 and GIII 14-I-605 strains showed replication rates and mean maximum titers at least 2.7-fold and 2.3-fold higher than those of GIV B254, respectively (Bonferroni posttest, p < 0.01). In Vero cells, the epidemic GETV strains caused more pronounced cytopathic effects in comparison to the GIV B254. Our findings suggest that higher virus replication competency that produces higher virus titers during infection may be the main determinant of virulence and epidemic potential of GETV.     

Structural Insights into Alphavirus Assembly Revealed by the Cryo-EM Structure of Getah Virus

Getah virus (GETV) is a member of the alphavirus genus, and it infects a variety of animal species, including horses, pigs, cattle, and foxes. Human infection with this virus has also been reported. The structure of GETV has not yet been determined. In this study, we report the cryo-EM structure of GETV at a resolution of 3.5 Å. This structure reveals conformational polymorphism of the envelope glycoproteins E1 and E2 at icosahedral 3-fold and quasi-3-fold axes, which is believed to be a necessary organization in forming a curvature surface of virions. In our density map, three extra densities are identified, one of which is believed a “pocket factor”; the other two are located by domain D of E2, and they may maintain the stability of E1/E2 heterodimers. We also identify three N-glycosylations at E1 N141, E2 N200, and E2 N262, which might be associated with receptor binding and membrane fusion. The resolving of the structure of GETV provides new insights into the structure and assembly of alphaviruses and lays a basis for studying the differences of biology and pathogenicity between arthritogenic and encephalitic alphaviruses.     

云南省2006-2010年狂犬病病毒N和G基因分子结构特征分析

目的了解云南省狂犬病毒的流行情况以及街毒株N、G基因结构特征,为有效控制狂犬病疫情提供初步科学依据。方法对云南省2006-2010年10株狂犬病街毒株N、G基因进行全基因克隆、测序,并与已知国内外代表毒株核苷酸及推导氨基酸进行序列比对及系统发育分析。结果序列分析表明所研究的10个云南毒株与Ⅱ、Ⅲ、Ⅳ、Ⅴ、Ⅵ、Ⅶ型代表毒株N、G基因核苷酸序列同源性分别为72.1%～78.5%、58.4%～71.0%,与基因Ⅰ型毒株序列同源性为85.2%～90.1%、82.7～99.6%,云南毒株之间核苷酸序列同源性为89.3%～99.2%、86.0～99.6%。云南10个毒株均为基因Ⅰ型和血清1型毒株,分为2个进化分支,除YNTC06与GXN119和HM88单独属于分支Ⅲ外,其余云南毒株均分布在分支Ⅰ上,且进一步分为3个小亚分支。遗传进化分析表明,各毒株氨基酸位点均存在不同程度的变异,其中YNTC06在360、369、375等多个NP抗原位点存在变异;云南毒株在GP330、333位毒力决定位点未发生变异,均为强毒株。结论云南狂犬病毒属于基因Ⅰ型,存在2个分支;云南狂犬病毒NP和GP存在特异性氨基酸变异位点,其基因结构及变异、亚组群划分与地理位置无相关性。     

Distribution of mosquitoes and mosquito-borne viruses along the China-Myanmar border in Yunnan Province

A total of 54,673 mosquitoes were collected at 11 sites located near the China-Myanmar border in the western part of Yunnan Province during July and August 2007. There were 29 species in 4 genera identified from the collections, including 12 species of Culex, 12 species of Anopheles, 3 species of Aedes, and 2 species of Armigeres. Culex tritaeniorhynchus Giles (67.9%, 37,119/54,673) and Anopheles sinensis Wiedemann (25.9%, 14,170/54,673) were the most abundant species in this investigation. Virus was isolated using BHK-21 and C6/36 cells from 22 of 510 mosquito pools. Isolates included Japanese encephalitis virus (JEV) and Getah virus (GETV), which were identified by serological and molecular methods. Twenty JEV strains were isolated from Cx. tritaeniorhynchus (15 isolates), An. sinensis (3 isolates), and Armigeres subalbatus Coquillett (2 isolates), and 2 GETV strains were isolated from Culex pseudovishnui Colless and Cx. tritaeniorhynchus. This study suggests that Ar. subalbatus is a potentially important local vector because of the high JEV infection ratio found in this species. Enzootic JEV transmission persists in this area and therefore, surveillance for human disease caused by JEV and GETV should be conducted in the region.     

基于灰色模型的全国肺结核疫情预测及分析

利用《中国疾病预防控制信息系统》获取2008—2015年全国肺结核疫情相关数据作为初始数据序列,使用灰色预测方法中的GM(1,1)模型对2016—2021年肺结核疫情相关数据进行预测,在此基础上使用自组织映射(self organizing maps,SOM)神经网络方法对预测结果进行聚类分析。依据灰色预测模型及2008—2015年肺结核疫情相关数据,预测得到2016—2021年全国及31个省(自治区、直辖市;不包括我国台湾、香港和澳门地区,下同)的发病数和发病率。根据预测结果,进一步利用SOM神经网络方法将全国31个省(自治区、直辖市)聚类成4个不同层次,重视程度从高到低依次分为第1类类中心序列(包括青海、西藏和新疆)、第2类类中心序列(包括黑龙江、湖南、广西、海南和贵州)、第3类类中心序列(包括辽宁、安徽、江西、河南、湖北、广东、重庆、四川、云南和陕西)和第4类类中心序列(包括北京、天津、河北、山西、内蒙古、吉林、上海、江苏、浙江、福建、山东、甘肃和宁夏)。研究认为,由灰色预测方法得出未来6年肺结核疫情呈下降趋势,可根据聚类所得的结果,针对不同类型的地区,适时调整防治策略,调配防治力量。     

立位心电图ST-T变化对器质性心脏病伴β-受体过敏综合征的诊断意义

目的　探讨立位心电图 ST- T变化在器质性心脏病伴 β-受体过敏综合征诊断中的意义。方法　对 35例健康者 (组 ) ,4 2例 β-受体过敏综合征 (组 ) ,34例器质性心脏病 (组 ) ,36例器质性心脏病伴 β-受体过敏综合征 (组 )患者的卧、立位心电图 ST- T波及心率进行对比研究 ,并观察心得安试验对其影响。结果　组 与组 ,组 与组 卧、立位心率增快程度不同 ,后者均分别大于前者 (P<0 .0 1) ;组 患者的卧位心电图 ST- T正常 ,转为立位时ST- T发生异常改变 (71.4 2 % ) ,或卧位心电图 ST- T异常 ,转为立位时 ST- T异常的程度与范围加重 (2 8.5 7% ) ;组 心电图 ST- T正常及组 原有 ST- T改变者分别在卧、立位对比时无明显变化 ;组 者立位时原异常 ST- T变化的程度与范围加重 (10 0 % ) ;卧位心得安试验使组 患者的心电图 ST- T改变完全恢复正常 ,组 无改善 ,组 轻度改善。结论　立位心电图 ST- T动态变化、心率增快程度及心得安试验 ,对器质性心脏病伴 β-受体过敏综合征的诊断、治疗指导具有重要意义