猪戊型肝炎病毒ORF3蛋白影响HepG2细胞核黄素代谢的lncRNA-mRNA调控网络的鉴定

为初步鉴定并挖掘出猪戊型肝炎病毒(Hepatitis E virus,HEV)ORF3蛋白影响HepG2细胞核黄素代谢信号通路的lncRNA-mRNA调控网络,本试验通过构建腺病毒过表达载体,制备高滴度过表达腺病毒,介导猪 HEV-ORF3在HepG2细胞中实现过表达。Western blotting检测ORF3蛋白过表达成功后,运用lncRNA高通量组学测序,筛选出差异表达的lncRNA并进行靶向差异基因预测,对lncRNA靶向差异基因进行GO功能和KEGG通路富集分析,初步鉴定出与核黄素代谢信号通路相关的lncRNA-mRNA调控网络。Western blotting结果显示,在约为12 ku处出现目的条带,说明成功实现腺病毒介导猪HEV-ORF3在HepG2细胞中过表达。lncRNA高通量组学测序结果显示,共发现102个显著差异表达lncRNAs表达量上调,80个显著差异表达lncRNAs表达量下调。GO功能和KEGG通路富集分析显示,初步挖掘出lncRNA(MSTRG.13995.2)和lncRNA(MSTRG.1960.1)可能是与核黄素代谢信号通路相关的显著差异表达lncRNA,分别通过顺式调控其靶向基因APC-5和FLAD1来影响核黄素代谢信号通路。本试验初步鉴定出lncRNA(MSTRG.13995.2)-APC5和lncRNA(MSTRG.1960.1)-FLAD1可能是影响HepG2细胞核黄素代谢信号通路的lncRNA-mRNA调控网络。     

上海地区多种动物戊型肝炎血清学调查

在上海地区收集血液样品(猪、牛、山羊、马、鸡、鸭、鸽、犬和部分珍禽等)964份,应用双抗原夹心酶联免疫法(DS-ELISA)检测抗HEV抗体。结果表明,猪抗HEV抗体的阳性率为72.18%(301/417),其中母猪阳性率最高,为82.53%(222/269),育成猪阳性率为52.54%(62/118),保育猪阳性率为50.00%(15/30);22个猪场均检测到抗HEV抗体阳性猪,阳性率高于60%的猪场有16个,占阳性猪场的72.72%。在牛、山羊、马、鸡、鸭、鸽等血清中也检测到了抗HEV抗体,阳性率分别为6.00%,24.00%,16.00%,1.90%,12.77%和4.44%,明显低于猪群的阳性率。在宠物犬中检测到了抗体阳性犬,阳性率为17.82%(18/101),说明犬对HEV易感。珍禽中野鸭、火烈鸟对HEV也有一定的敏感性,阳性率分别为6.60%(1/15)和10%(1/10),在孔雀和鸵鸟的血清标本中未检测到HEV抗体。猪群中戊型肝炎病毒存在较普遍,其他与人类密切相关的多种动物也对HEV敏感。     

流式细胞术检测TcR Vα24/Vβ11双阳性NKT细胞及其在HCV感染者中的应用

目的研究丙肝病毒感染者外周血NKT细胞含量与HCV感染状态之间的相关性.方法应用流式细胞术检测TcR Vα24/Vβ11双阳性NKT细胞.结果抗-HCV阳性且HCVRNA也阳性的病例NKT细胞数量明显低于抗-HCV阳性而HCV RNA阴性的病例（P〈0.01）,明显低于正常对照组（P〈0.05）.结论由于NKT细胞数量减少或活化受阻,造成HCV的持续感染,不利于病毒的清除;另一方面,由于肝内有HCV的存在,NKT细胞大量向肝内趋化,导致外周血的数量相对减少.     

Detection of HCV core protein,envelope glycoprotein and core-envelope fusion protein by baculovirus surface display

AIM: To detect the hepatitis C virus (HCV) at early stages by the technique of baculovirus surface display. METHODS: We constructed the recombinant baculoviruses vAc-Flag-HCV C-GP64, vAc-Flag-HCV E-GP64 and vAc-Flag-CE-GP64, which displayed HCV-related structural proteins by the technique of baculovirus surface display. RESULTS: Western blotting results showed that the core (C), envelope (E) and core-envelope (CE) proteins of HCV were expressed in Sf9 cells after the infection of recombinant baculoviruses. We also observed by confocal microscopy that the C, E and CE proteins of HCV presented and were anchored on the plasma membrane of Sf9 cells after infection. In addition, the results of immunogold electron microscopy demonstrated that the 3 recombinant baculoviruses displayed the C, E and CE proteins of HCV on the viral surface, respectively. To further define the role of these recombinant baculoviruses in detecting HCV at early stages, we applied time-resolved fluoroimmunoassay (TRFIA) to detect HCV samples after purification and concentration of the recombinant baculoviruses. The results showed that the positive rate of vAc-Flag-CE-GP64 was up to 80.2%. CONCLUSION: vAc-Flag-CE-GP64 is useful for detecting the hepatitis C virus at early stages.     

The levels of 2′, 5′ oligoadenylate synthetase,interleukin 2 and interleukin 12 in hepatitis B virus infection

AIM: In order to study the effect of endogenous interferon system and Th1 response modes on hepatitis B virus infection, the 2′, 5′ oligoadenylate synthetase (2-5OAS), IL-2 and IL-12 were selected as the research parameters. METHODS: The activity of 2-5OAS in peripheral blood mononeuclear cells was determined by sensitive radioenzymatic assay. IL-2 and IL-12 were determined by ELISA. RESULTS: Compared to normal control, the 2-5OAS, IL-2 or IL-12 were not significantly changed (P>0.05) in the asymptomatic HBsAg carricer group. The 2-5OAS, IL-2 and IL-12 were significantly up-regulated (P<0.01) in the group of acute hepatitis, but in the groups of chronic hepatitis, liver cirrhosis and hepatocellular carcinoma, the 2-5OAS, IL-2, IL-12 were significantly down-regulated (P<0.05). Moreover, with the progression of patient′s conditions and with the complications of liver cirrhosis and hepatocellular carcinoma, the 2-5OAS, IL-2 and IL-12 decreased progressively, the 2-5OAS, IL-2, IL-12 were the lowest in guoups of liver cirrhosis and hepatocellular carcinoma (vs each groups of chronic hepatitis, P<0.05). CONCLUSION: The endogenous interferon system and Th1 response are significantly alterable in the different period of hepatitis B virus infection and among the different clinical types. The cellular immunity plays an important role in recovery from HBV infection.     

中药复方治疗感染DHBV麻鸭的研究

本研究以人工感染鸭乙型肝炎病毒（ Ｄ Ｈ Ｂ Ｖ）的麻鸭为实验模型，用两组中药复方进行体内抗病毒实验研究。结果表明：１实验结束时，两个治疗组麻鸭的平均体重增长值显著高于对照组（ Ｐ＜ ０００１），而平均肝重显著低于对照组（ Ｐ＜ ０００１），肝系数也显著小于对照组（ Ｐ＜ ０００１）；２两个治疗组麻鸭随着服药时间的延长， Ｄ Ｈ Ｂ Ｖ 转阴率持续增加，实验结束时转阴率均分别显著高于对照组（２９２７％ 与３０５６％ ８１１％ ， Ｐ＜ ００５）；而且未出现 Ｄ Ｈ Ｂ Ｖ 转阴后又转阳现象。提示这两组中药复方不仅具有抗病毒的作用，而且药物安全、药效稳定，且有较好的治疗效果；建议试用于临床治疗乙型肝炎。     

草仙乙肝胶囊治疗慢性肝炎（湿邪困脾症）Ⅱ期临床研究

草仙乙肝胶囊是吉林省龙井制药厂根据南京中医学院附属医院治疗慢性乙型肝炎的经验方，经现代工艺加工研制而成的一种新.药该药具有清热解毒、健脾利湿之功效，主治慢性乙型肝炎，中医辨证属于湿邪困脾症．经临床试验，其效果显著，无毒副作用     

Seroprevalence of Hepatitis E Virus in Domestic Pigs and Wild Boars in Switzerland

Hepatitis E is considered an emerging human viral disease in industrialized countries. Studies from Switzerland report a human seroprevalence of hepatitis E virus (HEV) of 2.6–21%, a range lower than in adjacent European countries. The aim of this study was to determine whether HEV seroprevalence in domestic pigs and wild boars is also lower in Switzerland and whether it is increasing and thus indicating that this zoonotic viral infection is emerging. Serum samples collected from 2,001 pigs in 2006 and 2011 and from 303 wild boars from 2008 to 2012 were analysed by ELISA for the presence of HEV‐specific antibodies. Overall HEV seroprevalence was 58.1% in domestic pigs and 12.5% in wild boars. Prevalence in domestic pigs was significantly higher in 2006 than in 2011. In conclusion, HEV seroprevalence in domestic pigs and wild boars in Switzerland is comparable with the seroprevalence in other countries and not increasing. Therefore, prevalence of HEV in humans must be related to other factors than prevalence in pigs or wild boars.     

HBV衣壳蛋白装配抑制剂的研究进展

乙型肝炎病毒(HBV)是一种常见的嗜肝性病原体,全球约有4亿人感染,带来严重的社会问题和经济负担。虽然目前已有预防和治疗HBV的药物,但由于耐药性等问题不能彻底清除病毒感染,新药开发依然迫在眉睫。介绍了HBV的病毒结构、复制和感染过程,对现有治疗药物的优缺点进行了总结。对以衣壳蛋白装配为靶点的新药研究进展进行综述,旨在为今后开发抗HBV的新药提供参考。     

苦参注射液治疗乙型肝炎性肝硬化疗效及其作用机制研究

目的：观察苦参注射液对慢性乙型肝炎性肝硬化的临床效果及其作用机制.方法将58例慢性乙型肝炎性肝硬化患者随机分为观察组和对照组,在一般支持治疗的基础上,观察组加用苦参注射液,治疗8周后,比较两组肝功能、肝纤维化指标.结果观察组肝功能改善显著,肝纤维化减轻,与对照组比较差异均有统计学意义（P〈0.05）.结论在正常治疗基础上加用苦参注射液,有助于肝炎性肝硬化患者的肝功能恢复,有较好的临床效果.其作用机制可能是通过影响外周单核细胞IL-10表达、调节机体促炎抗炎平衡而实现的.