小鼠巨细胞病毒模型的建立

目的为了探讨巨细胞病毒的致病机理。方法４周龄Ｂａｌｂ／Ｃ小鼠腹腔内接种小鼠巨细胞病毒（ＭＣＭＶ）。结果导致小鼠急性感染期体重下降，生长迟缓，唾液腺肿胀以至于死亡。唾液腺中检出高滴度感染性病毒（２．０×１０５ＰＦＵ／ｍｌ）。在小鼠３Ｔ３／Ｓｗｉｓａｌｂｉｎｏ细胞单层上形成的空斑清晰，易判断计数、镜检组织切片可见脑神经原细胞胞浆内包涵体。结论小鼠巨细胞病毒模型的建立为抗－ＣＭＶ有效药物的筛选以及对ＣＭＶ感染的预防、治疗提供了资料。     

脑心肌炎病毒（EMCV）感染昆明小鼠模型的建立

目的：建立EMCV感染昆明小鼠模型。方法：6～8周龄昆明小鼠,雌雄各半,分为空白对照组和EMCV高(100TCID₅₀/20 g)、中(10 TCID₅₀/20 g)、低(1 TCID₅₀/20 g)剂量组。分别于EMCV感染5 d、7 d、9 d、11 d、13 d、15 d称重,眼眶采血后,断颈处死。取心、脑、肝和脾组织,计算各器官脏器指数;qRT-PCR检测心、脑、脾、肝和外周血病毒载量;qRT-PCR检测心和脑IL-1β、IL-6和TNF-α mRNA表达量;取心和脑,HE染色后观察组织病理损伤。结果：与空白对照组相比,高剂量的EMCV感染小鼠5 d时,能显著提高心脏指数(P<0.05)和极显著提高脾脏指数(P<0.01);qRT-PCR结果显示EMCV感染9 d时,小鼠心、脑、肝、脾和外周血病毒载量最高,中、低剂量EMCV感染小鼠各脏器中的病毒载量很低;高剂量的EMCV感染小鼠9 d时,心和脑IL-1β(P<0.01)、IL-6(P<0.001)和TNF-α(P<0.01)极显著升...     

女性淋病患者宫颈HCMV和HSV—Ⅱ感染PCR诊断分析

采用聚合酶健反应（PCR）技术对387例女性淋病患者进行宫颈HCMVDNA和HSV－ⅡDNA检测，并对比观察156例孕女宫颈HCMV和HSV－Ⅱ感染情况。结果表明女性淋病患者宫颈HCMVDNA阳性率为25．06％，HSV－ⅡDNA阳性率为15．24％；孕妇宫颈HCMVDNA阳性率为7．05％，HSV－ⅡDNA阳性率为0．64％。提示女性淋病患者多有非婚性接触史而发生HcMV和HSV－－Ⅱ感染。     

小鼠人偏肺病毒感染模型的建立

目的 建立人偏肺病毒(hMPV)感染小鼠模型,了解病毒肺内复制规律及所致病理改变,为hMPV感染免疫病理机制研究及新型防治手段开发奠定基础.方法 BALB/c小鼠经滴鼻感染荧光标记的重组hMPV,于感染后1、3、5、7、9、16 d处死小鼠并无菌获取肺组织用于病毒分离和病理检查,改良噬斑形成法检测hMPV滴度,RT-PCR法检测hMPV mRNA表达.结果 小鼠滴鼻感染hMPV后肺组织分离到病毒;肺组织病毒滴度在感染后5 d达到高峰(5.16±1.09)×105PFU/g,感染后第9大仍能检测到病毒(2.79±1.22)×102PFU/g;感染后16 d肺组织仍可检测到hMPV mRNA;病理改变在感染后3～7 d最明显,为典型的间质性肺炎改变.结论 hMPV感染BALB/c小鼠模型建立成功,可用于hMPV感染的免疫病理机制研究.     

抗磷脂综合征患者脑脊液单纯疱疹1型病毒和巨细胞病毒抗体阳性病例分析

目的:报道4例与抗磷脂综合征(APS)患者脑脊液有关病毒抗体的阳性结果。方法:检测4例确诊APS患者脑脊液中单纯疱疹1型病毒(HSV-1)和巨细胞病毒(CMV)抗体,结合临床及影像学资料,分析其与APS发病的关系。结果:4例患者脑脊液HSV-1-IgG、CMV-IgG均为阳性,HSV-1-IgM、CMV-IgM均为阴性;例1、例2血清HSV-1-IgG、CMV-IgG阳性,例3、例4阴性;例1、2、3抗心磷脂抗体(aCL)血清水平12RU/ml;例4检出抗β2糖蛋白1(β2GP1)抗体。结论:初步提示HSV-1和CMV感染与APS可能存在相关性。     

蚊媒刺叮对Balb/C小鼠感染登革Ⅱ型病毒的增强作用

登革热 登革出血热是我国重要的蚊媒病之一 ,白纹伊蚊和埃及伊蚊是主要媒介蚊种。本研究以Balb C小鼠为实验动物 ,探讨白纹伊蚊唾液对登革病毒感染宿主的增强作用。结果显示 ,在感染剂量相同的情况下 ,如果Balb C实验小鼠预先被一定数量的白纹伊蚊叮咬以后再皮下接种病毒 ,实验小鼠感染登革病毒的程度有所提高媒介蚊虫叮咬宿主吸血时会分泌唾液,蚊媒病毒随分泌的唾液传播给宿主,蚊虫唾液不仅是病毒传播的液体环境,还对病毒感染宿主有一定促进作用,通过影响宿主的免疫功能而起作用。蚊虫等吸血双翅目昆虫由于吸血时间短,被叮咬的宿主免疫系统可能来不及发生反应,所以以前推测它们对宿主免疫方面的作用可能比较小或没有(Champagne,1994)。近年研究发现能抑制或调节动物宿主免疫反应,影响宿主对虫媒病原体的免疫能力,如发现白蛉唾液中的舒血管物质能抑制宿主巨噬细胞功能,增加利什曼原虫的感染(TitusandRibeiro,1988;Theodosetal.,1993),之后相关的研究逐渐多起来(Bissonnetteetal.,1993;Crossetal.,1994)。对Cachevalley病毒感染传播研究发现只有通过蚊虫有效的叮咬Balbc实验小鼠才能产生病毒血症和相应的抗体,而仅皮下注射病毒则不能(Edwardsetal.,1998);对LaCross病毒的研究也发现同样的问题(Osorioeta     

蓝舌病毒Hbc株与肿瘤细胞相互作用的研究

蓝舌病毒(Bluetongue Virus，BTV)是呼肠孤病毒科(reoviridae)环状病毒属(Genus Orbivirus)的代表种，主要感染牛、羊及野生反刍动物，引起动物蓝舌病(Bluetongue disease)。中国动物蓝舌病毒湖北株(BTV-Hbc)是从湖北省分离得到的病毒株，为动物病毒，不感染人正常细胞，不引起人体任何疾病。但是它和人源的其它细胞，     

单纯疱疹病毒Ⅱ型和人乳头瘤病毒的感染与人工流产术后不孕症的关系

目的探讨性传播病毒和不孕症的关系。方法应用聚合酶链反应（ＰＣＲ）对６０例人工流产术后不孕症妇女和３９例正常妇女进行了生殖道单纯疱疹病毒Ｉ型（ＨＳＶ２）和人乳头瘤病毒（ＨＰＶ）的检测。结果不孕组和对照组ＨＳＶ２的阳性检出率分别是８０．０％和２５．５％，两组间有极显著性差异（Ｐ＜００１）；ＨＰＶ的阳性率分别是５３３％和３３３％，两组间无显著性差异（Ｐ＜０．０５）；ＨＳＶ和ＨＰＶ在两组中的混合感染的阳性率分别是４３．３％（２６／６０）和２３．１％（９／３９），两者有显著性差异（Ｐ＜０．０５）。表明ＨＳＶ２或ＨＳＶ２和ＨＰＶ的混合感染与人工流产术后不孕症有显著的相关性，很可能是不孕的原因之一。两组９９份标志中，ＨＳＶ２和ＨＰＶ混合感染的阳性率为３５．３５％，统计学分析表明，ＨＳＶ２和ＨＰＶ感染与不孕有极显著的相关性χ＝１２．５，Ｐ＜０．０１。结论ＨＳＶ２和ＨＰＶ的感染和不孕症相关     

人乳头瘤病毒、沙眼衣原体和单纯疱疹病毒Ⅱ型感染与宫颈疾病的相关性研究

目的 探讨人乳头瘤病毒(human papillomavirus,HPV)、沙眼衣原体(chlamydia trachomatis,CT)和单纯疱疹病毒Ⅱ型(herpes simplex virusⅡ,HSV-Ⅱ)感染与宫颈疾病的关系.方法 采用核酸分子快速杂交基因分型技术对598例宫颈脱落细胞进行HPV感染基因型分型测定,同时应用实时荧光定量PCR法(FQ-PCR)检测CT和HSV-Ⅱ两种病原体的感染情况.结果 HPV、CT和HSV-Ⅱ的阳性检出率分别为24.9％、9.2％和8.9％,且随着宫颈病变程度加重而逐渐增高,宫颈炎组和宫颈癌病变组三种病原体的感染率差异均有统计学意义.HPV感染以高危型为主(占77.9％),低危型为22.1％.HPV高危型和阴性组比较两种病毒的感染率有统计学差异(CT:x2=26.97,P＜0.05;HSV-Ⅱ∶x2=65.93,P＜0.05).各宫颈病变组均有多病毒混合感染情况出现,其中宫颈癌组混合感染率为22.2％.结论 HPV、CT和HSV-Ⅱ感染与宫颈癌的发生密切相关,多病原体混合感染会促进宫颈癌的发生.     

Application of shRNA-containing herpes simplex virus type 1 (HSV-1)-based gene therapy for HSV-2-induced genital herpes

HSV-1-based vectors have been widely used to achieve targeted delivery of genes into the nervous system. In the current study, we aim to use shRNA-containing HSV-1-based gene delivery system for the therapy of HSV-2 infection. Guinea pigs were infected intravaginally with HSV-2 and scored daily for 100 days for the severity of vaginal disease. HSV-2 shRNA-containing HSV-1 was applied intravaginally daily between 8 and 14 days after HSV-2 challenge. Delivery of HSV-2 shRNA-containing HSV-1 had no effect on the onset of disease and acute virus shedding in animals, but resulted in a significant reduction in both the cumulative recurrent lesion days and the number of days with recurrent disease. Around half of the animals in the HSV-2 shRNA group did not develop recurrent disease 100 days post HSV-2 infection. In conclusion, HSV-2 shRNA-containing HSV-1 particles are effective in reducing the recurrence of genital herpes caused by HSV-2.     

Immunocytochemical confirmation of oral and respiratory herpesvirus infection by the immunoperoxidase method

Recognition of diagnostic viral changes in general, and due to herpesvirus infection (HVI) in particular, from a variety of materials examined cytologically has added a new dimension to diagnostic cytology. Such recognition provides valuable information to the clinician when no infection is suspected. Previously, the diagnosis of HVI of the respiratory tract was only possible at postmortem, tissue culture, or diagnostic rise in antibody titer. This study describes our experience in the diagnosis of HVI of the respiratory tract initially suggested by sputum cytology and later confirmed by the immunoperoxidase method. The importance of sputum cytology in the diagnosis of HVI is emphasized.     

鼠逆转录病毒小鼠模型的建立

艾滋病（ＡＩＤＳ）的大流动，仍在全球迅速蔓延，极大地危害人类的生命及健康，临床迫切需要寻找新的抗人免疫缺陷病毒（ＨＩＶ）芗，小动物模型是发展新药的重要环节，我们引进Ｃ型鼠白血病病毒，建立了Ｂａｌｂ／Ｃ小鼠感染模型，观察了ＲＶＢ－３的感染特征。用ＡＩＤＳ首选药－－叠氮胸苷（ＡＺＴ）及黄芩提取品（ＳＢＴ），在该模型内进行实验治疗，两者在体内实验中均显示明显的抗病毒效果。     

Seroepidemiology of Herpes Simplex Virus and Restriction Endonuclease Cleavage Analysis of Herpes Simplex Virus Type 2 in Okinawa

A seroepidemiologic study of herpes simplex virus (HSV) in Okinawa was performed. A total of 423 serum samples were collected from all over Okinawa, and the positivity rate of antibody against HSV was measured using a passive hemagglutination method. The sero positive rate for HSV in age groups of over 40 years was 100%. Seven HSV type 2 (HSV 2) isolates were obtained in Okinawa, and DNA preparations from Vero cells infected with the isolates were analyzed using five restriction endonucleases: Bam HI, Hind III, Kpn I, Bgl II and Eco Rl. Variations in the genomic region were demonstrated in five of the isolates. Such variations have not been reported previously in HSV 2 in mainland Japan. This is the first report of a seroepidemiologic study of HSV and restriction endonuclease cleavage analysis of HSV 2 in Okinawa, is a subtropical island where HSV is endemic. Acta Pathol Jpn 41: 24–30, 1991.     

单纯疱疹病毒体外诱导特异性抗体应答的实验研究

用灭活的单纯疱疹病毒（Ｈｅｒｐｅｓｓｉｍｐｌｅｘｖｉｒｕｓ，ＨＳＶ１）为致敏原，体外诱导成人外周血淋巴细胞产生特异性抗体应答，特异性抗体诱生水平与血清抗体无相关性（Ｒ＝０．４５，Ｐ＞０．０５），抗体类型为ＩｇＧ。同法致敬新生儿淋巴细胞则不能诱生任何类型的特异抗体，表明此体外抗体应答属继发性免疫应答。特异性抗体应答有明显的ＨＳＶ１抗原剂量依赖性，且需要Ｔ、Ｂ细胞的相互作用。ＨＳＶ１体外致敏的实验研究，为探讨正常和疾病状态下体外特异性抗体应答和免疫调节提供一个有用的实验模型。     

Complement component C3b binds directly to purified glycoprotein C of herpes simplex virus types 1 and 2

Cells infected with herpes simplex virus type 1 (HSV-1), but not HSV-2, express on their surfaces a receptor for the complement component C3b. Receptor activity is markedly enhanced by treatment of the infected cells with neuraminidase. Employing a direct binding assay, consisting of purified HSV glycoproteins immobilized on nitrocellulose and iodinated C3b as a probe, we found that C3b binds directly to gC-1, as well as to gC-2, but not to gB or gD from either serotype. C3b binding was enhanced by treatment of gC-1 or gC-2 with neuraminidase. Endo F or endo H treatment of gC-1 had no effect on C3b binding. However, treatment of gC-2 with these endoglycosidases had a marked negative effect on C3b binding. These results suggest that N-linked oligosaccharides are involved in binding of C3b to gC-2, but not gC-1. Alternatively, removal of N-linked oligosaccharides from gC-2 might adversely affect polypeptide conformation. Glycoprotein C-2 also differs from gC-1 in its effects on the complement cascade. Whereas gC-1 accelerated the decay of the alternative pathway C3 convertase and impaired the efficiency of lysis by the components C5 through C9, gC-2 stabilized the active C3 convertase and had little effect on the late-acting components. The dissimilarity of gC-1 and gC-2 with regard to their effects on the complement cascade may have implications regarding the role of these glycoproteins in confronting the host immune response.     

单纯疱疹病毒胸苷激酶基因介导GCV治疗人膀胱癌的实验研究

探讨单纯疱疹病毒胸苷激酶(Herpes simplex virous thymidine kinase,HSV-TK)基因介导的丙氧鸟苷(Ganciclovir,GCV)系统对人膀胱癌细胞(T24)生长的抑制作用。利用电穿孔技术将携有HSV-TK基因的重组逆转录病毒载体(pLXSN-TK)导入包装细胞PA317,获得转染后的逆转录病毒包装细胞PA317／TK。将BALB／c裸鼠24只随机分为4组(每组6只),建立人T24裸鼠荷瘤模型。治疗1,2组分别瘤内注射病毒悬液或PA317／TK细胞后再经GCV处理。对照1,2组则分别瘤内注射等量生理盐水或：PA317／TK细胞后再经等量GCV或生理盐水处理。观察各组肿瘤体积和瘤重的变化;采用原位杂交技术分析HSV-TK基因在肿瘤组织中的表达;采用流式细胞仪技术(Flow cytometry,FCM)和末端脱氧核苷酸介导的X-dUTP缺口末端标记技术(Termininal deoxynucleofidyl trasferase-mediated dUTP nick end labelling technigue,TUNEL)检测膀胱癌细胞凋亡。结果表明：(1)逆转录病毒介导的HSV-TK基因在体内能转染T24细胞,联合运用GCV后,肿瘤的生长速度明显减慢;(2)HSV-TK／GCV治疗后膀胱癌细胞凋亡现象明显增强。以上结果提示T24荷瘤裸鼠内HSV-TK／GCV系统对T24细胞的生长有明显的抑制作用,且可能与细胞凋亡引起的“旁观者效应”有关。HSV-TK／GCV系统在人类膀胱癌治疗方面具有明显的临床运用前景。     

丙肝病毒NS5A蛋白对NS5B的RdRP活性影响的研究

丙型肝炎病毒 (Hepatitis C virus,HCV )非结构蛋白 (Nonstructural,NS) 5 A在 HCV基因组复制中的作用目前尚不清楚。本文研究 His- NS5 A对 NS5 B的 RNA依赖性 RNA酶 (Rd RP)活性的影响 ,以了解 NS5 A在HCV RNA复制中的作用。采用变性 -复性方法 ,纯化大肠杆菌表达的重组组氨酸 NS5 A融合蛋白。 GST结合洗脱实验 (GST pull- down assay)研究 NS5 A和 NS5 B是否结合。以不同的摩尔浓度比 ,将纯化的 NS5 B和 NS5 A蛋白混合 ,检测 NS5 A对 NS5 B的 Rd RP活性的影响。获得高得率的纯化 His- NS5 A蛋白。重组 NS5 A蛋白可在体外与NS5 B结合并抑制后者的 Rd RP活性。本研究报道了纯化重组 His- NS5 A蛋白的变性 -复性方法 ,结果显示纯化的重组 NS5 A在体外可与 NS5 B相互结合 ,并明显抑制 NS5 B Rd RP活性。提示了 HCV NS5 A在病毒复制中的可能作用。     

巢式PCR检测龈下菌斑中HCMV、EBV、HSV-1与慢性牙周炎活动性的关系

目的　建立临床检测龈下菌斑标本中人巨细胞病毒 (HCMV)、Epstein Barr病毒 (EBV)、单纯疱疹病毒 1型 (HSV 1 )巢式PCR方法 ,研究这 3种病毒与慢性牙周炎活动性的关系。方法　收集6 2例慢性牙周炎患者 (男性 2 7例、女性 35例 ,平均年龄 5 3岁 )的牙周炎活动部位、牙周炎静止部位的龈下菌斑 ,提取DNA后使用巢式PCR检测HCMV、EBV、HSV 1 ,比较分析其在同一病人不同部位的检出率。结果　牙周炎活动部位的HCMV检出率为 38.7%,EBV的检出率为 5 8%,HSV 1的检出率为30 .6 %,两种以上病毒合并感染的检出率为 4 0 .3%;牙周炎静止部位的HCMV检出率为 1 4 .5 %,EBV为 2 2 .6 %,HSV 1为 1 1 .3%,两种以上病毒合并感染的检出率为 1 1 .3%。这 3种病毒及其合并感染在牙周炎活动部位的检出率均高于牙周炎静止部位 ,差异有统计学意义 (P <0 .0 5 )。结论　提示HC MV、EBV、HSV 1与慢性牙周炎的活动性相关。     

Effect of Gardenia extract ZG on the adsorption quantity of herpes simplex virus type 1 （HSV-1）

To observe the effect of Gardenia extract ZG on the adsorption quantity of herpes simplex virus type 1 (HSV-1) so as to explore the mechanism of its antiviral activity, fluorescein isothiocyanate (FITC) was used as the fluorescent probe to label viruses and heparin sodium was used as control. Meanwhile , the effect of Gardenia extract ZG on the adsorption quantity on the surface of Hep-2 cells was determined by flow cytometry. It was demonstrated that adsorption of HSV-1 on the surface of Hep-2 cells exhibited the character of saturation and specificity and heparin sodium could prevent attachment of viruses on these cells. These results are in accord with those reported previously. It was also proved that the manner of drug-use prior to adsorption or simultaneous use of drug and adsorption was better than adsorption prior to drug-use, and the inhibition rates of the former and latter manner were 84. 76% and 82.92% respectively. Three manners of drug-use with Gardenia extract ZG were all effective to reduce the adsorption quantity of viruses, especially the manner of simultaneous use of drug and adsorption with an adsorption inhibition rate of 68.46% . From the above observation, it is apparent that the mechanism of anti-viral activity of Gardenia extract ZG may be via several steps involved in the HSV-1 adsorption.