青天葵抗甲、乙型流感病毒作用研究

目的研究青天葵在体外抗甲、乙型流感病毒株作用。方法选用狗肾细胞培养法考察青天葵不同极性部位对甲型(FM1)、乙型(昆40B)流感病毒的作用。结果青天葵水溶性部位对甲型流感病毒FM1有抑制作用。结论青天葵具有体外抗甲型流感病毒作用。     

Viral infections in the mouth

Oral hairy leukoplakia (OHL) and Kaposi's sarcoma (KS) are commonly encountered in the HIV-infected patient. A unique feature of OHL is non-cytolytic high level of replication of Epstein–Barr virus (EBV) in the glossal epithelium. The expression of viral-encoded anti-apoptotic proteins concomitant to replicative proteins probably underlies this phenomenon. The question of whether OHL arises from activation of EBV latent in the tongue, or from superinfection by endogenous EBV shed via non-glossal sites or by exogenous EBV remains unresolved. Human herpesvirus 8 (HHV8) is now seen as necessary but not sufficient cause of KS. Expression of HHV8-encoded oncogenic proteins in endothelial cells probably explains the aberrant proliferation of these cells in KS lesions. Studies into why KS is so commonly observed at the palate in HIV-infected patients may provide important clues to its pathogenesis.     

HBV_2患者血清HBsAg的氨基酸组成分析

将２例ＨＢＶ＿２患者血清ＨＢｓＡｇ纯化，分析了ＨＢＶ＿２的ＨＢｓＡｇ氨基酸组成，并与分子量为６３０１９８道尔顿ＨＢＶ＿１的ＨＢｓＡｇ三种多肽氨基醚组成比较。实验发现，ＨＢＶ＿２与ＨＢＶ＿１的ＨＢｓＡｇ的氨基醚组成有较大区别，且区别指数均在１３以上。提示ＨＢＶ＿１和ＨＢＶ＿２的ＨＢｓＡｇ在一级结构上存在较大差异；ＨＢＶ＿２可能系一新的乙型肝炎病毒，也可能系由于ＨＢＶ＿１Ｓ区核苷酸突变所致的变异株。     

Epstein-Barr virus latent and replicative gene expression in oral hairy leukoplakia

Oral hairy leukoplakia is an epithelial lesion of the tongue associated with productive infection by Epstein-Barr virus (EBV). However, no data concerning the pattern of EBV latent gene expression have been reported, and it remains unresolved whether true latent infection occurs in basal cell layers of oral hairy leukoplakia. We have studied six cases of oral hairy leukoplakia using monoclonal antibody immunohistology for EBV latent--EB nuclear antigen (EBNA) 1, EBNA 2 and latent membrane protein 1 (LMP 1); immediate-early (BZLF1); and replicative (EA, VCA, MA) proteins, and for the EBV-receptor (CD21 antigen). EBV DNA was demonstrated by nucleic acid in situ hybridization. Mid- to upper-zone keratinocytes contained EBV DNA and co-expressed EBNA 1, EBNA 2 (5 of 6 cases), LMP 1, BZLF1 protein, EA, VCA and MA. No EBV genome or gene expression could be demonstrated in basal or parabasal cells. Spinous keratinocytes were labelled by anti-CD21 antibodies HB5 and B2, but did not express the EBV-receptor as defined by reactivity with OKB7. The co-expression of latent and replicative infection-associated antigens is striking, indicating possible functional roles for latent proteins during the productive cycle. Our results suggest that oral hairy leukoplakia is caused by repeated direct infection of upper epithelial cells with virus from saliva or adjacent replicatively infected cells, rather than by a latent EBV infection of basal epithelial cells with a differentiation-dependent switch to productive infection as previously proposed.     

粘病毒抵抗基因-1启动子88位点G/T多态性影响乙型肝炎病毒感染的自然转归

目的探讨乙型肝炎病毒（HBV）自限感染和慢性感染与粘病毒抵抗基因-1（MxA）启动子的-88位点G／T单核苷酸多态性的关系。方法收集100例抗-HBs和抗-HBc阳性的HBV自限感染者和340例慢性感染者的外周全血，提出基因组DNA；采用竞争分化聚合酶链反应技术为基础的方法进行MxA-88G／T基因分型；采用单因素Odds ratio和x^2检验等方法进行统计学分析。结果MxA-88G／G基因型（低表达型）检出率为50．2％（221／440），T／T基因型（高表达型）检出率为5．5％（24／440），G／T杂合型检出率为44．3％（195／440）。与慢性感染患者相比，自限感染患者携带较低的G／G基因型（41．0％与52．9%，P〈0．05）、G等位基因（62．5％与75．3％，P〈0．01）和较高的T／T基因型（16．0％与2．4%，P〈0．01）、T等位基因（37．5％与24．7％，P〈0．01），而两者之间的G／T杂合型差异无统计学意义。结论MxA-88G／T基因型能在一定程度上影响HBV感染的自然转归，有望成为临床上HBV感染转归的预测指标。     

HBV前C/C基因与绿色荧光蛋白基因融合表达载体的构建及在HEK293细胞中的表达

目的 :构建含HBV前C C基因与绿色荧光蛋白基因的融合表达载体 ,并在真核细胞中表达。方法 :用PCR法扩增含 1.3倍HBVayw型全基因组真核表达载体pHBV1.3的前C C基因片断 ,应用含绿色荧光蛋白基因的真核表达质粒构建融合表达载体 ,采用脂质体介导方法将其转染到HEK2 93细胞 ,并用荧光显微镜及ELISA法检测融合蛋白的表达。结果 :经PCR及酶切鉴定 ,证实成功构建了含HBV前C C基因的真核表达重组体pEGFP -HB VC ,显微镜下观察及ELISA法证实在HEK2 93细胞中表达了相应融合蛋白。结论 :构建的重组表达载体能在真核细胞中表达目的蛋白与GFP的双功能融合蛋白 ,适合于针对HBV前C C区的相关研究。     

重组2型腺相关病毒-转化生长因子β3对实验性肝纤维化大鼠肝脏组织病理学和Ⅰ型胶原表达的影响

Objective To investgate the effects of TGF β 3 on rat hepatic fibrosis. Methods The TGF β 3 cDNA was cloned into rAAV2 vector. Rats were randomly divided into four groups: normal control group, model group, negative control group and TGF β 3 group. Hepatic fibrosis was induced by hypodermic injection of 40% CCI4. Recombinant AAV2-TGF β 3 viral particles were injected via the vena caudalis one week before CCh treatment. Rats were executed 8 weeks after CCI4 treatment, global histological change was observed after HE staining, the distribution of the collagen fibers was observed after masson staining, his-tochemistry was done to observe the expression of collagen Ⅰ; The positive area rate of the collagen fibers and the average optical rate of collagen Ⅰ were quantified. Results HE staining indicated that collagen fibers were reduced in the TGF β 3 group. Masson staining shown that the collagen fibers were distributed around the blood vessel, in the portal area and disse space. Compared to the model group (13.2%±2.2%) and negative control group (12.3%±1.5%), the collagen fibers in liver tissues of TGF β3 group (7.7% ± 1.5%) were significantly decreased (q = 9.456, P < 0.01; q = 8.217, P < 0.01). Histochemistry indicated that the collagen fibers of liver tissues of TGF 15 3 group (0.185±0.033) were significantly higher than those in the model group (0.252±0.042) and the negative control group (0.230±0.029), (q = 6.228, P < 0.01; q = 4.346, P < 0.01). Conclusion TGF β 3 alleviates the damage to hepatic cell and the level offibrosis in CCI4 treated rats and inhibits the expression of collagen Ⅰ.     

Feline Immunodeficiency Virus Neuropathogenesis: From Cats to Calcium

Invasion of human immunodeficiency virus (HIV) into the central and peripheral nervous system produces a wide range of neurological symptoms, which continue to persist even with adequate therapeutic suppression of the systemic viremia. The development of therapies designed to prevent the neurological complications of HIV require a detailed understanding of the mechanisms of virus penetration into the nervous system, infection, and subsequent neuropathogenesis. These processes, however, are difficult to study in humans. The identification of animal lentiviruses similar to HIV has provided useful models of HIV infection that have greatly facilitated these efforts. This review summarizes contributions made from in vitro and in vivo studies on the infectious and pathological interactions of feline immunodeficiency virus (FIV) with the nervous system. In vivo studies on FIV have provided insights into the natural progression of CNS disease as well as the contribution of various risk factors. In vitro studies have contributed to our understanding of immune cell trafficking, CNS infection and neuropathogenesis. Together, these studies have made unique contributions to our understanding of (1) lentiviral interactions at the blood–cerebrospinal fluid (CSF) barrier within the choroid plexus, (2) early FIV invasion and pathogenesis in the brain, and (3) lentiviral effects on intracellular calcium deregulation and neuronal dysfunction. The ability to combine in vitro and in vivo studies on FIV offers enormous potential to explore neuropathogenic mechanisms and generate information necessary for the development of effective therapeutic interventions.     

JC病毒样颗粒可直接转运进入细胞核

目的探讨JC病毒(JCV)病毒样颗粒(VLP)是否可以直接转运进入细胞核。方法心用JCV主要外壳蛋白VP1体外表达、重组VIP，在其表面标记异硫氰基荧光素(FTTC)，同时在其内部包裹荧光染料Cy3，感染培养的HeLa细胞和SVG细胞，荧光显微镜观察VLP入核转运。结果HeLa和SVG细胞感染包裹Cy3的FTTC-VLP时，FTTC与Cy3同时出现于细胞核内相同部位；而感染FTTC—VP1与Cy3混合物时，FTTC虽可在细胞核内检测到，但Cy3信号几乎消失。包裹Cy3的VIP用SDSPAGE展开，荧光显像后行考马斯亮蓝(CBB)染色，发现Cy3和VLP移行至不同部位，证明Cy3不能与VP1结合，提示VLP以完整的颗粒形式转运进入细胞核。应用包裹外源性DNA的VLP感染培养的HeLa和SVG细胞，发现包裹的DNA在细胞浆和细胞核内均可检测到，提示JCV入核过程与VIP相同。结论VLP可以不经裂解直接转运进入细胞核，JCV入核转运可能与VLP相同。     

乙型肝炎病毒免疫逃避株表面抗原决定簇编码区的序列分析

应用聚合酶链反应（ＰＣＲ）产物直接测序，在国内首次发现１例持续高滴度乙型肝炎表面抗原（ＨＢｓＡｇ）和抗一ＨＢｓ共存者携带的乙型肝炎病毒ＤＮＡ第５３２位碱基Ａ被Ｇ取代，推导其“ａ”决定簇中第１２６位苏氨酸被丙氨酸取代，提示该株“ａ”决定簇第一个结构环疏水性增加，由此可能导致其抗原性改变，而诱发免疫逃避株形成。