聚合酶链反应鉴定丙型肝炎病毒母婴传播的状态及其意义

采用多种方法，动态检测了１１例丙型肝炎病毒（ＨＣＶ）感染的孕妇所生的婴儿血抗－ＨＣＶ和ＨＣＶＲＮＡ。发现用合成肽酶联免疫吸附试验（ｓｐＥＬＩＳＡ）检测婴儿抗－ＨＣＶ阳性率（２３．５２％）显著低于第二代重组抗原ＥＬＩＳＡ（２ｎｄＥＬＩＳＡ）（４１．１８％）（Ｐ＜０．０５）；用２ｎｄＥＬＩＳＡ检测，６例婴儿脐血和静脉血抗－ＨＣＶ阳性，５例持续１～５月阴转，１例阳性持续１３个月。经重组免疫印迹试验（ＲＩＢＡ）鉴定，４例阳性，２例可疑阳性。用逆转录聚合酶链反应（ＲＴ－ＰＣＲ）检测ＨＣＶＲＮＡ，５例阳性，３例于生后１～６个月自然阴转，２例持续阳性分别达９个月和１３个月。提示检测抗－ＨＣＶ判断ＨＣＶ母婴传播的状态受到婴儿抗－ＨＣＶ产生水平低下、母体抗－ＨＣＶ的被动输入和不同检测方法的影响，用ＲＴ－ＰＣＲ检测ＨＣＶＲＮＡ是判断母婴传播更可靠的指标。     

丙型肝炎病毒基因型与血清HCV RNA含量关系及与干扰素应答的研究

目的探讨丙型肝炎病毒（ＨＣＶ）基因型与血清ＨＣＶＲＮＡ含量的关系及其对干扰素应答的影响。方法应用定量荧光ＰＣＲ（Ａｍｐｌｉｓｅｎｓｏｒ－ＰＣＲ）技术检测了１３５例不同基因型ＨＣＶ感染的慢性丙型肝炎患者血清ＨＣＶＲＮＡ含量，另对其中７７例进行干扰素治疗并随访１２个月以上。结果ＨＣＶ－Ⅱ型感染血清ＨＣＶＲＮＡ水平（１０７．８±３．４拷贝／ｍｌ）显著高于ＨＣＶ－Ⅲ型感染（１０６．３±２．５拷贝／ｍｌ）（Ｐ＜０．０１），Ⅲ型感染的应答率（７／１３，５３．８％）显著高于Ⅱ型感染（２０／６４，３１．３％）（Ｐ＜０．０５）。应答组治疗前血清ＨＣＶＲＮＡ含量（１０６．８±２．７拷贝／ｍｌ）显著低于无应答组（１０８．３±３．２拷贝／ｍｌ）（Ｐ＜０．０１）。ＨＣＶＲＮＡ含量低于１０６．５拷贝／ｍｌ者，无论何种型别ＨＣＶ感染均应答较好，而ＨＣＶＲＮＡ高于１０８．０拷贝／ｍｌ者则应答极差。结论ＨＣＶ基因型及病毒血症水平是预测干扰素疗效的重要因素，且后者比前者意义更大。Ⅱ型感染病毒血症水平较高可能是影响其疗效的原因之一。     

831名健康青年庚型肝炎病毒和人免疫缺陷病毒感染的调查

目的了解我国健康青年中庚型肝炎病毒（ＨＧＶ）和人免疫缺陷病毒的感染情况。方法采用酶联免疫法（ＥＩＡ）检测６省８３１名健康青年血清中的ＨＧＶ和ＨＩＶ抗体，对抗－ＨＧＶＩｇＧ阳性的血清再用逆转录－巢式聚合酶链反应（ＲＴ－ＰＣＲ）检测ＨＧＶＲＮＡ。结果发现抗－ＨＣＶＩｇＧ阳性率为２５３％（２１／８３１），２１例阳性者中ＨＧＶＲＮＡ阳性８例，两者符合率为３８１％；抗－ＨＩＶ均阴性。结论我国健康青年人群中确实存在ＨＧＶ感染。     

抗HSV糖蛋白单克隆抗体可变区基因的克隆和序列测定

选用具有动物体内高保护作用的抗ＨＳＶ－１／ＨＳＶ－２型共同性糖蛋白Ｃ（ｇＣ）单克隆抗体１Ａ１２，从其杂交瘤细胞中提取总ＲＮＡ，以Ｏｌｉｇｏ（ｄＴ）１５为引物反转录合成ｃＤＮＡ，用一对鼠抗体通用ＶＨ引物和一对Ｖ引物进行ＰＣＲ，扩增出１Ａ１２ＶＨ和１Ａ１２ＶＬ基因，并分别克隆入ｐＵＣ１８质粒中，序列分析结果表明，１Ａ１２ＶＨ基因由３３６ｂｐ组成，编码１１２个氨基酸，隶属于小鼠重链第３组亚组（Ｄ）；１Ａ     

中国株庚型肝炎病毒(HGV)感染猕猴的实验研究

应用０．５ｍｌ含中国株庚型肝炎病毒（ＨＧＶ）的血清接种５只猕猴，５只猕猴于接种后１周血清ＨＧＶＲＮＡ均阳转；ＨＧＶＲＮＡ滴度在猴体内可高达１∶１０５，较接种血清中ＨＧＶＲＮＡ滴度（１∶１０２）高出许多，提示ＨＧＶ在猕猴体内复制。其中４只猕猴血清抗－ＨＧＶ阳转；４只出现ＡＬＴ异常。后用其中１只猕猴感染后４５天的血清给另２只猕猴接种，该２只猴也出现血清ＨＧＶＲＮＡ和抗－ＨＧＶ阳转及ＡＬＴ异常。本研究表明，中国猕猴有可能作为ＨＧＶ感染的动物模型。     

丙型肝炎病毒感染者中抗病毒IgM检测的意义

建立了抗ＨＣＶＩｇＭ间接ＥＬＩＳＡ方法，并用之检测ＨＣＶ不同感染人群。抗ＨＣＶＩｇＭ在不同感染人群中的检出率变化很大。急性输血后丙型肝炎患者检出率可高达９３．８％，而正常献血员中可低至０．６８％。比较抗ＨＣＶＩｇＭ和抗ＬｇＧ阳性中ＨＣＶＲＮＡ的检测结果发现，抗ＩｇＭ阳性者中，不同ＨＣＶ感染人群的ＨＣＶＲＮＡ检出率很高（９３．０％～１００％）；而ＩｇＧ阳性者中ＨＣＶＲＮＡ的检出率变化很大（３７．５％～９３．８％）。在４３例血液透析者中抗ＩｇＭ与ＨＣＶＲＮＡ检测的一致性为８３．７％；抗ＩｇＭ与抗ＩｇＧ检测的一致性为８８．４％，结果提示：（１）抗ＨＣＶＩｇＭ与ＨＣＶ活跃复制有关，（２）抗ＨＣＶＩｇＭ与抗ＨＣＶＩｇＧ检出不完全一致。因此，临床检测抗ＨＣＶＩｇＭ有其特殊意义。     

输血后丙型肝炎病毒感染的前瞻性研究

对６４例受血者进行了半年前瞻性调查，发生输血后丙型肝炎（ＰＴ－ＨＣ）８例，亚临床（ＰＴ－ＨＣ）１例，丙型肝炎病毒（ＨＣＶ）隐性感染３例，ＨＣＶ总感染率为１８７５％，丙氨酸转氨酶（ＡＬＴ）首次异常时间为输血后２８～９１（５１９±２０９）天；抗－ＨＣＶ首次阳转为输血后２３～７６（４２４±１５９）天。发生巨细胞病毒（ＣＭＶ）感染２例，病原待定的非乙、非丙ＡＬＴ异常者５例。     

831名健康青年庚型肝炎病毒和人免疫缺陷病毒感染？…

目的 了解我国健康青年中庚型肝炎病毒（ＨＧＶ）和人免疫缺陷病毒的感染情况。方法 采用酶联免疫法（ＥＩＡ）检测６省８３１名健康青年血清中的ＨＧＶ和ＨＩＶ抗体，对抗－ＨＧＶＩｇＧ阳性的血清再用逆转录－巢式聚合酶链反应（ＲＴ－ＰＣＲ）检测ＨＧＶＲＮＡ。结果 发现抗－ＨＧＶ ＩｇＧ阳性率为２．５３％（２１／８３１），２１例阳性者中ＨＧＶ ＲＮＡ阳性８例，两者符合率为３８．１％；抗－ＨＩＶ均阴性。结论 我     

合肥地区输血后慢性肝炎HCV基因分型研究

目的：探讨合肥地区输血后慢性肝炎ＨＣＶ感染的基因型。方法：用逆转录聚合酶链反应（ＲＴＰＣＲ）检测ＨＣＶＲＮＡ,对ＨＣＶＲＮＡ阳性标本用型特异引物ＰＣＲ进行ＨＣＶ分型。结果：６３ 例患者中检出ＨＣＶ　ＲＮＡ５４ 例,阳性率为８５．７％ ;其中Ⅱ型４６ 例（８５．２％）,Ⅲ型６ 例（１１．１％）,Ⅱ／Ⅲ型２ 例（３．７％）。结论：提示合肥地区ＨＣＶ感染大多数属Ⅱ型。     

利用双扩增聚合酶链反应检测非甲非乙型肝炎患者血清中丙型肝炎病毒RNA

利用丙型肝炎病毒（ＨＣＶ）５’－端序列合成两对引物，建立了灵敏、特异的ＨＣＶＲＮＡ双扩增聚合酶链反应检测方法。用此方法及第二代Ａｂｂｏｔｔ酶联抗－ＨＣＶ检测试剂盒，检测了４４例非甲非乙型肝炎患者血清及１０名抗－ＨＣＶ阴性健康人。在４４例患者中，４１例（９３％）ＨＣＶＲＮＡ阳性，３６例（８２％）抗－ＨＣＶ阳性，３３例（７５％）ＨＣＶＲＮＡ、抗－ＨＣＶ全部阳性。３例ＨＣＶＲＮＡ阴性，但抗－ＨＣＶ阳性，另外，有８例抗－ＨＣＶ阴性，ＨＣＶＲＮＡ阳性。１０名健康人ＨＣＶＲＮＡ均为阴性。结果表明，大部分（９２％）抗－ＨＣＶ阳性患者带有ＨＣＶ，但为了检测所有病毒血症患者，抗－ＨＣＶ检测是不够的，利用双扩增ＰＣＲ方法检测ＨＣＶＲＮＡ对于抗－ＨＣＶ阴性患者的诊断是非常有用的。     

Hepatitis C virus infects T cells and affects interferon-gamma signaling in T cell lines

It has been reported that hepatitis C virus (HCV) may infect and replicate in human T cells, particularly in perihepatic lymph nodes, but the extent and consequence of T-cell infection in patients is unclear. This study is conducted to characterize the parameters and functional consequences of HCV infection in T lymphocytes. By using a lymphotropic HCV strain, we showed that HCV could infect T cell lines (Molt-4 and Jurkat cells) in vitro. Both positive- and negative-strand HCV RNA were detected for several weeks after infection. Viral proteins could also be detected by immunofluorescence studies. Moreover, infectious HCV particles were produced from Molt-4 cell cultures, and could be used to infect na?ve T cell lines. HCV could also infect human primary CD4+ T cells, particularly na?ve (CD45RA+CD45RO-) CD4+ cells, in culture. The amounts of STAT-1 and phosphorylated STAT-1 proteins in the infected Molt-4 cells were significantly less than those in uninfected cultures, suggesting the possibility of defect in interferon-gamma signaling. Indeed, T-bet and STAT-1 mRNA levels after interferon-gamma stimulation in infected Molt-4 were suppressed. In conclusion, HCV could infect and transiently replicate in T cells and that HCV replication suppressed the IFN-gamma/STAT-1/T-bet signaling due to the reduction of STAT-1 and inhibition of its activation (phosphorylation).     

Ⅰ型单纯疱疹病毒特异性IgG、IgM及中和抗体的消长动态

本文采用酶联免疫吸附试验(ELISA)间接法、固相抗 IgM ELISA及微量中和试验对 HSV-1感染家兔特异性 IgG、IgM 及中和抗体的消长动态进行观察。结果表明,特异性IgM 在初次感染后第4天可检出,第8～15天达峰值,第66天完全消失,再感染后则检测不出。特异性 IgG 及中和抗体在初次感染后第 8～15天出现,第22天达峰值,检出时间长达 186天以上,再感染后滴度骤升50～100倍。     

Evolution of hepatitis C virus (HCV) infection acquired at birth

Objectives Our aim was to analyze the evolution of HCV infection in children infected at birth. Methods Between September 1994 and December 1998 we analyzed in a prospective study 8 children born of anti-HCV and HCV RNA positive women. Each baby was controlled at birth, every 3 months during the first year of life, and then every 6 months searching for anti-HCV antibodies (ELISA 3, RIBA 2-3), HCV RNA (RT PCR), ALT and viral genotype. Results Viral RNA was detectable in the first 3 months of life in all babies (100%) and remained positive during the follow-up. Viral genotypes were the same for mothers and their children. In 6 babies (75%) ALT remained pathologic during follow-up. Conclusions HCV infection in children usually has an asymptomatic outcome; the infection has chronic features in the majority of cases.     

Isolated core antibody hepatitis B in sub-Saharan African immigrants

Chronic hepatitis B virus (HBV) infection is a major health problem in sub-Saharan Africa, where prevalence is > or =8%, and is increasingly seen in African immigrants to developed countries. A retrospective audit of the medical records of 383 immigrants from sub-Saharan Africa attending the infectious diseases clinics at the Royal Melbourne Hospital was performed from 2003 to 2006. The HBV, human immunodeficiency virus (HIV) and hepatitis C virus (HCV) serological results are reported, with a focus on the isolated core antibody HBV pattern (detection of anti-HBc without detection of HBsAg or anti-HBs). Two-thirds (118/174, 68%) of those tested had evidence of HBV infection with detectable anti-HBc. Chronic HBV infection (serum HBsAg detected) was identified in 38/174 (22%) and resolved HBV infection (both serum anti-HBs and anti-HBc detected) in 45/174 (26%). The isolated core antibody pattern was identified in 35/174 (20%), of whom only 1/35 (3%) had detectable serum HBV DNA on PCR testing, indicating occult chronic HBV (OCHB). Only 8/56 (14%) patients with negative anti-HBc had serological evidence of vaccination (serum anti-HBs detected). HIV infection was detected in 26/223 (12%). HCV antibodies were detected in 10/241 (4%), of whom 8 (80%) had detectable HCV RNA. Viral co-infection was detected in only 2/131 (1.5%) patients tested for all three viruses. The isolated core antibody HBV pattern was common among sub-Saharan African patients in our study. These patients require assessment for OCHB infection and monitoring for complications of HBV.     

The relationship between viral RNA, myelin-specific mRNAs, and demyelination in central nervous system disease during Theiler's virus infection.

The DA strain of Theiler's murine encephalomyelitis virus (DAV) causes a chronic demyelinating disease in susceptible mouse strains. To elucidate the pathogenesis of DAV-induced demyelination, the authors investigated the spatial and chronologic relationship between virus (antigen and RNA), myelin-specific mRNAs, and demyelination in DAV-infected mice using immunohistochemistry, in situ hybridization, and slot blot hybridization analyses. In spinal cord white matter, viral RNA was detected easily in ventral root entry zones 1 to 2 weeks after infection. Viral RNA increased to maximum levels by 4 weeks after infection, which was associated with inflammation and mild demyelination. At 8 to 12 weeks after infection, when demyelination became most extensive, viral RNA was significantly decreased. Demyelination did not chronologically or spatially parallel the presence of viral RNA within the spinal cord. Decrease of myelin-specific mRNAs, including myelin-basic protein and proteolipid protein mRNAs, was observed within the demyelinating lesions with or without detectable viral RNA. These results indicate that a viral infection of white matter in the early phase of the infection initiates spinal cord disease leading to demyelination, but later an ongoing immunopathologic process contributes to the presence of extensive demyelination.     

Up-regulation of ERK and p38 MAPK signaling pathways by hepatitis C virus E2 envelope protein in human T lymphoma cell line

Hepatitis C virus (HCV) infection correlates with human immune disorders characterized by abnormal activation and proliferation of lymphocytes. Interaction of HCV major envelope protein E2 with susceptible cells occurs at an early stage of the viral infection. HCV tropism for susceptible cells may elicit cellular signaling events implicated in the viral pathogenicity, and E2 protein is known to be responsible for the tropism. We documented previously that HCV E2 protein was capable of activating extracellular signal-regulated kinase (ERK) in human hepatoma Huh-7 cells. Here, ERK and p38 mitogen-activated protein kinase (MAPK) signaling pathways were investigated in human T lymphoma cell line Molt-4 in response to HCV E2 protein. Binding of HCV E2 protein to Molt-4 cells was detectable, and such interaction was a determinant for recognition and delivery of the E2 signal to intracellular pathways. Activation of ERK and p38 MAPK was specifically induced following the HCV E2-cell interaction. CD81 and low-density lipoprotein receptor (LDLR), proposed cellular receptors for HCV, were expressed naturally on Molt-4 cells. CD81 and LDLR were shown to mediate HCV E2-induced activation of ERK and p38 MAPK. In CD81-deficient U937 cells, levels of ERK and p38 MAPK activation and cell proliferation induced by HCV E2 protein were lower than those in Molt-4 cells. Furthermore, cell proliferation and secretion of interferon-gamma and interleukin-10 by Molt-4 cells were promoted by HCV E2 protein. Therefore, ERK and p38 MAPK signaling pathways were up-regulated by HCV E2 protein without synergetic stimulation, which was accompanied by alterations of cell behavior.     

约氏疟原虫感染小鼠的巨噬细胞表面吞噬相关分子的研究

为探讨致死型约氏疟原虫(Plasmodium yoelii 17XL)感染抵抗型DBA/2小鼠的脾巨噬细胞发挥吞噬功能的作用机制,本试验利用Giemsa薄血膜染色,光学显微镜计数红细胞感染率,观察巨噬细胞的吞噬功能;采用酶联免疫吸附法(ELISA)检测小鼠血清中抗P.y.17XL特异性IgG抗体水平;采用流式细胞技术(FACS)动态检测脾巨噬细胞表面膜分子CD36、CD64表达水平。结果表明,DBA/2小鼠红细胞感染率于感染后第7天高达31.87%,约第15天自愈;感染小鼠脾巨噬细胞吞噬能力于感染后第5天开始增强,至第10天吞噬率高达95%,随后维持于高水平;巨噬细胞表面分子CD36于感染后第3天开始升高,至第10天达到峰值;CD64表达水平于感染后第5天开始升高,随后持续维持于高水平;小鼠血清中抗P.y.17XL特异性IgG抗体水平在感染后第10天开始出现有意义的升高。结果显示,在P.y.17XL感染过程中,巨噬细胞通过表面分子CD36和CD64分别介导的非调理性和调理性吞噬方式杀伤疟原虫,提示巨噬细胞是DBA/2小鼠发挥抗疟保护性免疫的重要效应细胞。     

A kinetic analysis of the expression of mast cell protease mRNA in the intestines of Nippostrongylus brasiliensis -infected rats

To study the kinetics and the phenotype of the mast cells (MC) arising during infection with the nematode Nippostrongylus brasiliensis, monospecific cDNA probes for nine different MC proteases were used in a Northern blot analysis of RNA from the small intestine of infected rats. The expression was analyzed at four individual time points during infection, day 0 (before infection), and days 7, 12 and 16 post infection. A dramatic increase in mRNA for rat mast cell protease (RMCP)-2, the major mucosal MC protease in the rat, was observed, beginning around day 7 after infection and peaking around day 12. At day 16 the expression was already beginning to decline. An almost identical pattern of mRNA expression was detected for the RMCP-8 subfamily of rat MC proteases (RMCP-8, −9 and −10) and for two additional rat serine proteases, the chymases RMCP-3 and −4. No simultaneous increase in the proteases known to be expressed preferentially by mature connective tissue MC (RMCP-1, −6 and −7) was observed. This is consistent with our finding that the expansion of MC in the intestines of parasite-infected animals was limited, almost exclusively, to the mucosal MC population. However, a minor increase in RMCP-5 and MC carboxypeptidase A (CPA) mRNA was detected at day 12 after infection, suggesting a derivation of mucosal MC from an expanding RMCP-5- and CPA-positive population of MC precursors.     

Cytotoxic activity of monocytes againstToxoplasma gondii in acute,chronic and reactivated murine toxoplasmosis

The phagocytic activity and cytotoxicity of peripheral blood monocytes (against toxoplasma tachyzoites) was studied in acute, chronic and reactivated toxoplasma infected Swiss albino mice. During acute infection, a low phygocytic activity was observed on the 4th day post infection (dpi) (P<0.01) and a low monocyte cytotoxicity was noticed after the 2nd dpi (P<0.01) which further decreased till the 8th dpi. In contrast, both the parameters were significantly increased during chronic infection. Increase in monocytic cytotoxicity was manifested on the 3rd dpi (P<0.001) whereas phagocytosis showed an increase on the 12th dpi (P<0.05). The reactivated group showed no change in both the parameters when compared with the control immunosuppressed group (P>0.05).