中蜂囊状幼虫病病毒核酸和多肽的研究

中蜂囊状幼虫病病毒(CSBV)核酸经聚丙烯酰胺凝胶电泳(PAGE)和琼脂糖凝胶电泳(AGE)分析,表明为单链RNA,易于降解;CSBV结构多肽经SDS-PAGE分析,有3个多肽,其分子量分别为27,000、29,000、39,000道尔顿。此病毒RNA和多肽与意蜂囊状幼虫病病毒(SBV)有差异。     

小麦丛矮病毒核酸的研究

小麦丛矮病毒(Wheat Rosette Stunt Virus.以下简称WRSV)的核酸可以通过SDS-尿素处理和苯酚抽提的方法分离获得。琼脂糖凝胶电泳测得核酸的分子量约为4.4×10~6道尔顿。核酸温度熔解曲线的测定证明它是一个单链的RNA。在电子显微镜下可观察到两种长度的核酸分子,一种长度约为3.5～4.0μ,相当于碱基数为14,000～16,000,另一种约为其一半;并能看到RNA的一些二级结构。     

黑曲霉病毒的形态和特性及其在细胞内的表现

从产生糖化酶的黑曲霉(Aspergillus niger)菌株中分离到一种等轴对称、衣壳表面有突起、内含双链RNA的病毒颗粒。病毒颗柱在电镜下直径为28—33nm,呈六面体晶格排列。病毒在蔗糖密度梯度离心中有三个分部,分析超离心所得沉降系数为161S,118S和94S。病毒在凝胶电泳中为一条带,在SDS-聚丙烯酰胺凝胶电泳中外壳蛋白分子量为90,000、82,000、76,000、52,0000、42,000 道尔顿。提取的病毒与其抗血清在免疫双扩散实验中出现一条沉淀线。病毒核酸有5个组分,与聚肌苷：聚胞苷[poly(1)：poly?]抗血清反应中出现一条沉淀线。在早期菌丝细胞超薄切片中,病毒颗粒多近似球状紧密聚集,外被以膜结构,聚生或散生于胞质中;后期菌丝细胞中病毒颗粒则多散生于胞质中。     

草菇病毒——一种新的食用菌ds-RNA病毒

自草菇(Folvariella~olvacea)子实体中分离到一种等轴对称直径为35rim左右的病毒颗粒。病毒样品经琼脂糖电泳显示一条区带,并具有典型核蛋白的紫外吸收光谱。最大吸收为E257,最低吸收为E230,A260／280=1．96。经SDS-不连续聚丙烯酰胺凝胶电泳解离病毒,出现一条主要衣壳多肽的电泳带,分子量为60,000道尔顿。病毒核酸的最大吸收在258nm,最低吸收在232nn,A260/280=2．0,琼脂糖电泳为一个组分。     

体外培养的红白血病细胞中Friend病毒合成的研究

Friend 白血病毒(FLV)是有双层套膜的C 型 RNA 肿瘤病毒,由受感染的细胞芽生。成熟的病毒颗粒呈圆形,直经约100毫微米,电子密度深的拟核含 RNA 和蛋白质,RNA 单链,约占病毒的1%。成熟的小鼠白血病病毒有类似鸟类髓细胞白血病病毒基因组的结构,两个35S 亚单位约为3×10~6道尔顿,RNA 分子量为10—12×10~6道尔顿。这类 RNA 肿瘤病毒(Oncorna 病毒)都含有反转录酶,它的合成受病毒基因组中 pol 基因的控制。基因组     

女贞的一种小分子闭环RNA

女贞(Ligustrum Cornpactum)叶片的核酸提取物,经聚丙烯酰胺凝胶电泳(PAGE),出现两条低分子量的核酸带,DNase 1及RNase A处理证实两条核酸带均为RNA,初步测得迁移率较小的RNA的分子量约为0.7×10~5道尔顿,5＇末端标记,双相及双方向PAGE分析证实,该RNA是存在于女贞叶片细胞内的一种小分子闭环RNA(简称psc RNA),不同离子强度下的RNase A处理揭示,psc RNA分子内有大量碱基互补结构,从点杂交结果推断psc RNA的一级结构中,不含绝大多数闭环类病毒RNA所共同具有的中心保守区段,在本实验的条件下pscRNA不能感染爪哇三七,女贞叶片汁液和按照提取,纯化病毒粒子的程序制备的溶液。在电镜下均没有观察到病毒颗粒,结果表明,女贞叶片细胞中存在一种无蛋白质外壳包被的、具有次量分子内碱基互辅结构的共价闭合环状RNA分子,在其它两种植物中,亦初步检测到此类闭环RNA分子,讨论了此种闭环RNA分子的意义。     

兔的一种新病毒 II．一株兔出血症病毒的某些理化性质的研究

采用氯仿、聚乙二醇。硫酸葡聚糖钠盐二相系统和蔗糖密度梯度离心法,从患病兔肝组织中提取、纯化病毒。该病毒粒子无囊膜,呈20面体对称,直径一般为33—37 nm。其三角剖分数T=3,共有32个子粒,每一子粒为中空的、外径为9 nm左右的圆形轮廓。经蔗糖密度梯度离心后,可获得沉降系数为166s的病毒颗粒,这种颗拉具有很强的感染性。经SDS—PAGE测得病毒粒子有四种多肽,分子量分别为66．4k、65．Ok、63．5k、41．Ok。二苯胺试验,吖啶橙染色、甲醛试验及热变性曲线表明,该病毒是单链DNA病毒。电镜下经甲酰胺法展层的病毒核酸呈单链线状,分子量平均为2．1×106道尔顿。此病毒的上述性质类似于细小病毒。 53s中空的圆形子粒,有很强的血凝性,而对照组铁蛋白和同一条件下所制备正常兔肝 成份无凝血特性,说明53s的9nm粒子应为病毒外壳蛋白子粒。     

米曲霉中的一种新病毒与其它真菌病毒的血清型比较

自α-淀粉酶生产菌米曲霉Aspergillus oryzae 3800中找到直径20～24nm的等轴病毒颗粒(AoV),病毒具有典型核蛋白吸收峰,最大E258,最小E242,E260/280=1.36,沉降系数为45S,一个电泳组分,毒粒对氯仿敏感,细胞内含量低,病毒核酸电泳一个组分,地衣酚反应阳性,SDS-不连续聚丙烯酰胺凝胶电泳解离病毒得到分子量为17,800道尔顿的一个主要衣壳多肽。 AoV与小麦全蚀病菌病毒(GgV)的几个衣壳多肽及分子量不同的分离物间有共同的抗原性,而与产黄青霉病毒和同属的黑曲霉病毒不同,这是真菌寄主分类地位较远的病毒间有共同抗原性的不常见例子。     

慢性蜜蜂麻痹病病毒核酸与蛋白质组分的研究

从自然感染的意大利麻痹病蜂(APis mellifera)头部,经二次差速离心与蔗糖梯度离心获得纯化的慢性蜜蜂麻痹病病毒(CBPV)。纯化的CBPV制备物感染正常蜜蜂,4天后出现典型的麻痹症状,接着死亡,平均死亡率分别为95％与100％。SDS-聚丙烯酰胺凝胶电泳分析,二次差速离心初步纯化的病毒制备物含有多条蛋白带,而蔗糖密度梯度纯化的病毒制备物仅含有单一的多肽带。5％、7.5％与10％SDS-聚丙烯酰胺凝胶电泳分析,均检测出一种病毒蛋白质,分子量大约为24,200道尔顿,而且不同凝胶浓度检测的蛋白质分子量相近。慢性蜜蜂麻痹病病毒核酸也用SDS-聚丙烯酰胺凝胶电泳分析,结果表明,凝胶中有5条带,对核酸酶敏感,证明该病毒含有5个单股RNA组分。对慢性蜜蜂麻痹病病毒的基因组结构进行了讨论。     

小鹅瘟病毒核酸链型与结构蛋白分析

采用简易方法从鹅胚中分离纯化的小鹅瘟病毒(GPV)样品相当纯净,以致在电镜下呈晶格排列。用负染色技术观察病毒的形态结构,病毒直径为18～20nm,无囊膜。病毒颗粒含有4种结构蛋白,其分子量分别为88、78、66和51kd。根据病毒的核酸链型分析,小鹅瘟病毒含有单链核酸。此结果支持了小鹅瘟病毒属于细小病毒属的结论。     

一种包装AAV2/5的重组单纯疱疹病毒的构建

为了得到一种可以包装AAV2/5和表达绿色荧光蛋白的重组单纯疱疹病毒,设计并构建了一个由AAV2rep基因和AAV5cap基因嵌合而成的rep2cap5基因,然后,利用一套携带HSV1基因组的粘粒系统(cos6、cos28、cos14、cos56、cos48),将rep2cap5基因插入cos6粘粒上HSV1基因组片段的UL2基因中,而将EGFP的表达单位插入cos56粘粒上HSV1基因组片段的UL44基因中,用这2个重组粘粒与其它3个粘粒(cos14、cos28、cos48)共转染BHK-21细胞获得了重组病毒HSV1-r2c5-EGFP并进行了空斑纯化。HSV1-r2c5-EGFP病毒能够在BHK-21细胞连续传代,并且可以观察到几乎所有的感染细胞都能产生绿色荧光。用PCR方法以及Southern杂交方法表明所获得的HSV1-r2c5-EGFP中携带有rep2cap5基因,用HSV1-r2c5-EGFP感染携带报告基因LacZ的AAV载体细胞株,获得了具有感染性的重组AAV2/5-LacZ。结果表明,所获得的重组单纯疱疹病毒HSV1-r2c5-EGFP可提供AAV2/5载体包装所需的全部辅助功能,是一种能简便、高效制备重组AAV2/5病毒的通用性辅助病毒。     

Polypeptide composition of flacherie viruses of the silkworm

The purified flacherie viruses of the silkworm, Bombyx mori, (FVS I, FVS II, FVS III, and FVS IV) were iodinated by using chloramine-T. The iodinated FVSes were purified by sucrose density gradient centrifugation or 2.4% polyacrylamide gel electrophoresis. FVS IV was found in the sedimentation analysis of FVS I, FVS II, and FVS IV. Electrophoretic patterns of FVS IV showed that it was a mixture of components having identical mobilities with FVS I, EVS IIa, and FVS IIb. FVS IV was a decomposed particle of FVS I, FVS II, and/or FVS III. All of these particles contained three polypeptides with molecular weights of about 51,000, 31,000, and 12,000 daltons. FVS I composed of six polypeptides with molecular weights of 67,000, 51,000, 39,000, 31,000, 14,000, and 12,000 daltons. The maturation process of FVS I was discussed and was suggested as the following process, FVS IIb→FVS IIa→FVS I. It is not clear whether FVS III is an intermediate for FVS IIa to convert into FVS I, or FVS III is a decomposed particle of FVS I.     

RNA viruses in the sea

Viruses are ubiquitous in the sea and appear to outnumber all other forms of marine life by at least an order of magnitude. Through selective infection, viruses influence nutrient cycling, community structure, and evolution in the ocean. Over the past 20 years we have learned a great deal about the diversity and ecology of the viruses that constitute the marine virioplankton, but until recently the emphasis has been on DNA viruses. Along with expanding knowledge about RNA viruses that infect important marine animals, recent isolations of RNA viruses that infect single-celled eukaryotes and molecular analyses of the RNA virioplankton have revealed that marine RNA viruses are novel, widespread, and genetically diverse. Discoveries in marine RNA virology are broadening our understanding of the biology, ecology, and evolution of viruses, and the epidemiology of viral diseases, but there is still much that we need to learn about the ecology and diversity of RNA viruses before we can fully appreciate their contributions to the dynamics of marine ecosystems. As a step toward making sense of how RNA viruses contribute to the extraordinary viral diversity in the sea, we summarize in this review what is currently known about RNA viruses that infect marine organisms.     

Evidence for plant viruses in the region of Argentina Islands, Antarctica

This work focused on the assessment of plant virus occurrence among primitive and higher plants in the Antarctic region. Sampling occurred during two seasons (2004/5 and 2005/6) at the Ukrainian Antarctic Station 'Academician Vernadskiy' positioned on Argentina Islands. Collected plant samples of four moss genera (Polytrichum, Plagiatecium, Sanionia and Barbilophozia) and one higher monocot plant species, Deschampsia antarctica, were further subjected to enzyme-linked immunosorbent assay to test for the presence of common plant viruses. Surprisingly, samples of Barbilophozia and Polytrichum mosses were found to contain antigens of viruses from the genus Tobamovirus, Tobacco mosaic virus and Cucumber green mottle mosaic virus, which normally parasitize angiosperms. By contrast, samples of the monocot Deschampsia antarctica were positive for viruses typically infecting dicots: Cucumber green mottle mosaic virus, Cucumber mosaic virus and Tomato spotted wilt virus. Serological data for Deschampsia antarctica were supported in part by transmission electron microscopy observations and bioassay results. The results demonstrate comparatively high diversity of plant viruses detected in Antarctica; the results also raise questions of virus specificity and host susceptibility, as the detected viruses normally infect dicotyledonous plants. However, the means of plant virus emergence in the region remain elusive and are discussed.     

寨卡病毒及其疫苗研究

寨卡病毒与黄热病毒、登革热病毒、日本脑炎病毒、西尼罗病毒等都属于蚊媒传播的黄病毒属病毒。寨卡病毒分离于1947年,但由于分布区域有限,所致寨卡热症状较轻,很长一段时间并没有引起太多的关注。最近一些年,特别是2015年后,巴西的寨卡疫情暴发及其与新生儿小头畸形的关联,引起了全球越来越多的关注。疫苗是应对寨卡疫情的重要手段,目前全球有30余个机构在进行寨卡病毒疫苗的研发。本文综述了寨卡病毒的生物学、流行病学、临床特征以及当前不同类型寨卡病毒疫苗研发现状,同时对其他几种黄病毒属病毒批准和临床阶段疫苗情况进行了概述,以为相关研究人员提供参考。     

Early onset of virus infection and up-regulation of cytokines in mice treated with cadmium and manganese

A substantial database indicates that a large number of environmental pollutants, chemicals and therapeutic agents to which organisms are exposed cause immunotoxicity. The suppression of immune functions may cause increased susceptibility of the host to a variety of microbial pathogens potentially resulting in a life-threatening state. Evaluation of the immunotoxic potential of chemical xenobiotics is of great concern and, therefore, we have investigated the impact of exposure of inorganic metals, specifically cadmium (Cd) and manganese (Mn) on Encephalomyocarditis virus (EMCV), Semliki Forest virus (SFV), and Venezuelan Equine Encephalitis virus (VEEV) infection. Pretreatment with a single, oral dose of Cd or Mn increased the susceptibility of mice to a sub-lethal infection of these viruses as observed by increased severity of symptoms and mortality compared to untreated controls. An early onset of virus infection was found in brains of Cd and Mn treated animals. Histopathological observations of the brain indicate evidence of inflammation and greater tissue pathology in Cd-or Mn-exposed mice compared to control animals. Meningitis and vascular congestion was seen in virus infected mice in all the metal treated groups, and further, the perivascular inflammation appeared earlier in treated mice compared to control. Encephalitis was maximum in Cd pretreated mice. Widespread environmental contamination of metals and the potential for their exposure and subsequent infection of humans or animals is indicative that further studies of these and all other metals are important to understand the effect of environmental pollution on human health.     

应用斑点法检测植物病毒的研究

应用斑点法检测了病叶粗汁液中的芜菁花叶病毒(TuMV)、大豆花叶病毒(sMV)和黄瓜花叶病毒(CMV),病叶粗汁液可被检测的最大稀释度分别为1:5120、1:2560和1:1280。提纯的大豆花叶病毒和黄瓜花叶病毒可检测的最低限量分别为1.7ng和1.2ng。以牛血清白蛋白、吐温和聚乙烯吡咯啉酮作封闭液,均可获得满意的结果。应用斑点法检测芜菁花叶病毒和大豆花叶病毒时,其抗血清稀释1:500倍可获得满意效果,稀释2000倍仍可用于检测。     

肝炎病毒与EB病毒重叠感染

为探讨肝炎病毒（ＨＶ）与ＥＢ病毒（ＥＢＶ）重叠感染的状况和后果,我们用免疫酶法对１５４例各型病毒性肝炎患者作了ＥＢＶＩｇＡ抗体检测。结果发现,急性肝炎、慢性轻度肝炎、慢性中度肝炎、肝炎肝硬化、慢性重型肝炎和原发性肝癌ＶＧＡ－ＩｇＡ抗体的阳性率分别为２４．０％、３０．０％、５３．３％、６３．３％、４０．０％和７２．７％,与健康人（５．３％）比较,有非常显著升高（Ｐ＜０．０１）;原发性肝癌又较急性肝炎和慢性轻度肝炎高,并有非常显著意义差异（Ｐ＜０．０１）。ＨＢＶ和ＨＡＶ＋ＨＢＶ感染者比较,前者又较后者低（Ｐ＜０．０１）。重叠感染者的临床表现均为“肝炎型”,未见咽炎、腺热、胃肠、肺炎、肾炎、神经等类型。重叠感染者的ＣＤ＋３及ＣＤ＋４Ｔ细胞下降,ＣＤ＋８Ｔ细胞及ＩｇＧ,ＩｇＭ升高,与健康人比较差异非常显著意义（Ｐ＜０．０１）。结果提示：ＨＶ感染,不仅因免疫失调易感ＥＢＶ,又可因重叠感染而进一步使免疫功能失调;对病毒性肝炎的处理应强调免疫调节治疗。