乙型肝炎病毒基因型的临床研究进展

乙型病毒性肝炎是世界范围的严重公共卫生问题之一。在3．7亿多乙型肝炎病毒携带者中，中国有近1．3亿。乙型肝炎是我国最常见的病毒性传染病，人群流行率为67％，慢性乙型肝炎患者有3000万，20％-40％将发展成慢性肝病。每年花费的医疗费用和保健费用高达500亿元。乙型肝炎病毒(HBV)已经被分成不同的血清亚型，后来又以核苷酸异源性为基础分     

新型抗乙型肝炎病毒药物的研究进展

目前HBV感染仍是全球性的公共卫生问题,全世界有高达4亿的慢性HBV感染者,我国约占10％.干扰素(IFN)及核苷(酸)类似物[nucleos(t)ide analogue,NUCs]的抗病毒疗效在大多数地区已得到认可.在抗病毒治疗方面虽有重大突破,但现有治疗方法却很少能达到病毒清除的效果,IFN不良反应大,而NUCs疗程长,易耐药.所以探索新型抗病毒药物是乙型肝炎防治领域的一大热点.随着对HBV生活史的深入了解及HBV感染系统的建立,在新型抗病毒药物研制方面取得很大进展.     

聚乙二醇干扰素α-2a治疗慢性丙型肝炎病毒／乙型肝炎病毒共感染临床疗效

目的研究慢性丙型肝炎（CHC）与丙型肝炎病毒/乙型肝炎病毒（HCV/HBV）共感染者用聚乙二醇干扰素α-2a（PEG-IFNα-2a）抗病毒治疗疗效的观察。方法治疗前后定期采集31例CHC、30例HCV/HBV共感染者的外周血,检测HCV RNA、HBV DNA和肝脏纤维化指标。结果治疗后CHC患者血清中HCV RNA含量51.6%〈102拷贝/ml,HCV/HBV共感染者HCV、HBV下降均不理想。结论 PEG-IFNα-2a抗病毒治疗HCV/HBV共感染者疗效较单纯丙型肝炎患者差,但对HBV、HCV有抑制作用,并部分改善机体免疫功能及肝功能。     

乙型肝炎病毒相关肝硬化的抗病毒治疗

慢性乙型肝炎（CHB）患者每年约有2．1％～6．0％进展为肝硬化。年龄、病毒复制程度、乙型肝炎病毒（HBV）基因型、合并丙型肝炎病毒（HCV）感染等都影响HBV感染后进展至肝硬化的速率。而进展至肝硬化后HBV仍可持续存在，血清乙型肝炎e抗原（HBeAg）或HBV DNA阳性，这些患者还处在进展至肝硬化失代偿和肝癌（HCC）的危险中，只有抗病毒治疗，即清除或持续抑制HBV，才能减少这些并发症的发生，延长生存率，提高生活质量。     

慢性乙型肝炎病毒合并丙型肝炎病毒感染伴2型糖尿病1例

一、病例资料 患者男性,63岁.广东湛江人,汉族,退休教师.因反复疲乏、纳差、尿黄15年余于2009年4月27日就诊.患者15年前无明显诱因出现疲乏,食欲不振,身目黄染、尿黄,曾到广东省人民医院就诊,检查发现肝功能异常,丙型肝炎抗体为阳性,诊断为"急性丙型肝炎",给予护肝治疗1月余,症状缓解.     

乙型肝炎病毒基因变异及其临床意义

HBV变异不仅可在慢性感染过程中自然发生,亦可于应用药物或接种疫苗后出现.病毒发生变异后常引起其生物学特征的改变,不仅其自身复制能力可能受到影响,同时可引发对抗病毒药物的耐药及导致严重的不良后果,这给乙型肝炎的预防、诊断和治疗等带来一系列新的问题.因此,加强对HBV变异特征的研究,具有重要的生物学和临床意义.     

乙型肝炎病毒嗜肝结合位点与受体的研究进展

众多学者对HBV嗜肝性机制得出共识:HBV膜蛋白直接与肝细胞膜上相应的受体结合,继而膜融合,通过胞饮作用内吞入胞;或者HBV与细胞外液中某一中介分子结合形成复合体,再与肝细胞膜上其相应的受体结合入胞.但目前仍不能确定膜受体及中介分子是什么.现就近年来HBV嗜肝机制研究综述如下.     

α-干扰素治疗成人乙型肝炎病毒相关肾炎的疗效观察

目的观察α-干扰素治疗成人乙型肝炎病毒(hepatitisBvirus,HBV)相关肾炎的疗效。方法对1996-09~2003-03上海交通大学附属瑞金医院9例HBV相关肾炎患者予以α-干扰素治疗,300×104U隔日治疗,疗程6~9个月,观察治疗前后蛋白尿、血清HBV标志物和分支链DNA(branchedDNA,bDNA)等情况的变化。结果经治疗,患者尿蛋白排泄量显著减少(P<0·05);9例患者中,8例患者蛋白尿有不同程度的减少,24h尿蛋白减少量为0·55~9·50g,其中3例蛋白尿消失,4例肾病综合征缓解。9例患者乙肝表面抗原(HBsAg)均未转阴,6例乙肝e抗原(HBeAg)阳性者3例转阴。9例患者检测了血清HBVbDNA,8例患者bDNA下降伴蛋白尿好转,1例bDNA无好转伴持续大量蛋白尿。α-干扰素治疗成人乙肝病毒相关肾炎总有效率为78%,无明显不良反应。结论抗病毒药物α-干扰素治疗成人HBV相关肾炎具有一定效果。治疗有效者蛋白尿减少,体内病毒复制下降,由此证实病毒复制在HBV相关肾炎中的作用。     

脂质体干扰素的体外抗乙型肝炎病毒效应

本文采用２．２．１５细胞进行脂质体干扰素与干扰素的体外抗乙型肝炎病毒实验，用ＥＬＩＳＡ法测ＨＢｅＡｇ，用反向血凝法及ＥＬＩＳＡ法测ＨＢｓＡｇ，用斑 交法测ＨＢＶ ＤＮＡ，结果证实，脂质体ＩＦＮ体外对ＨＢｅＡｇ的抑制率较ＩＦＮ高，能较好地抑制ＨＢＶ ＤＮＡ，但与ＩＦＮ一样，对ＨＢｓＡｇ无抑制作用。     

血清乙型肝炎病毒载量与抗病毒药物疗效

为了提高α-干扰素(IFN)及拉米夫定(LAM)两种抗病毒药物的抗病毒治疗应答率,我们对乙型肝炎病毒(HBV)不同血清含量的病人分组治疗,发现各组疗效有明显差异。     

乙型肝炎病毒X区核苷酸序列变异的检测

目的：了解乙型肝炎患者血清中乙型肝炎病毒（HBV）DNA X区核苷酸序列的变异情况。方法：采用聚合酶链反应（PCR）扩增24例乙型肝炎患者血清HBV DNA X区产物，并直接测序进行分析。结果：24例患者血清中HBV DNA X区都有程度不等的点突变（2－15），11例同时具有nt1762(A→T),nt1764(G→A）发生变异，8例患者同时在nt1636-nt1741几处位点发生变异。结论：HBV DNA X区核苷酸位于nt1762,nt1764双位变异与HBeAg阴性表型有关。     

Characterization of hepatitis B virus X gene quasispecies complexity in mono-infection and hepatitis delta virus superinfection

Hepatitis delta virus(HDV) seems to strongly suppress hepatitis B virus(HBV)replication, although little is known about the mechanism of this interaction. Both these viruses show a dynamic distribution of mutants, resulting in viral quasispecies. Next-generation sequencing is a viable approach for analyzing the composition of these mutant spectra. As the regulatory hepatitis B X protein(HBx) is essential for HBV replication, determination of HBV X gene(HBX)quasispecies complexity in HBV/HDV infection compared to HBV monoinfection may provide information on the interactions between these two viruses.AIM To compare HBV quasispecies complexity in the HBX 5' region between chronic hepatitis delta(CHD) and chronic HBV mono-infected patients.METHODS Twenty-four untreated patients were included: 7/24(29.2%) with HBeAgnegative chronic HBV infection(CI, previously termed inactive carriers), 8/24(33.3%) with HBeAg-negative chronic hepatitis B(CHB) and 9/24(37.5%) with CHD. A serum sample from each patient was first tested for HBV DNA levels.The HBX 5' region [nucleotides(nt) 1255-1611] was then PCR-amplified for subsequent next-generation sequencing(MiSeq, Illumina, United States). HBV quasispecies complexity in the region analyzed was evaluated using incidencebased indices(number of haplotypes and number of mutations), abundancebased indices(Hill numbers of order 1 and 2), and functional indices(mutation frequency and nucleotide diversity). We also evaluated the pattern of nucleotide changes to investigate which of them could be the cause of the quasispecies complexity.RESULTS CHB patients showed higher median HBV-DNA levels [5.4 logIU/mL,interquartile range(IQR) 3.5-7.9] than CHD(3.4 logIU/mL, IQR 3-7.6)(P = n.s.)or CI(3.2 logIU/mL, IQR 2.3-3.5)(P < 0.01) patients. The incidence and abundance indices indicated that HBV quasispecies complexity was significantly greater in CI than CHB. A similar trend was observed in CHD patients, although only Hill numbers of order 2 showed statistically significant differences(CHB2.81, IQR 1.11-4.57 vs CHD 8.87, 6.56-11.18, P = 0.038). There were no significant differences in the functional indices, but CI and CHD patients also showed a trend towards greater complexity than CHB. No differences were found for any HBV quasispecies complexity indices between CHD and CI patients. G-to-A and C-to-T nucleotide changes, characteristic of APOBEC3 G, were higher in CHD and CI than in CHB in genotype A haplotypes, but not in genotype D. The proportion of nt G-to-A vs A-to-G changes and C-to-T vs T-to-C changes in genotype A and D haplotypes in CHD patients showed no significant differences. In CHB and CI the results of these comparisons were dependent on HBV genotype.CONCLUSION The lower-replication CHD and CI groups show a trend to higher quasispecies complexity than the higher-replication CHB group. The mechanisms associated with this greater complexity require elucidation.     

与乙型肝炎病毒感染相关的易感或拮抗基因的研究进展

乙型肝炎病毒(HBV)感染是世界范围的严重公共卫生问题之一.HBV感染后,机体对病毒的清除能力以及疾病的进展和不同临床转归,除了与病毒因素和环境因素有关外,在很大程度上取决于个体间基因组的差异.本文就近年来与乙型肝炎病毒感染相关的易感或拮抗基因的研究进展及研究中存在的问题作一综述,主要包括影响机体免疫应答的基因、人类白细胞抗原(HLA)、肿瘤坏死因子(TNF)、白细胞介素(IL)、干扰素(IFN)等,同时对这一领域的研究前景作一展望.     

HBV感染自然史研究进展

病毒复制与宿主免疫之间的动态平衡在HBV感染自然史进展和发病机制中起重要作用。多数免疫能力正常的成人感染HBV后呈自限性,而在婴幼儿则多发展成为慢性HBV感染。慢性HBV感染分为4期：免疫耐受期、HBeAg阳性慢性肝炎期、非复制的HBsAg携带期和HBeAg阴性慢性肝炎期。HBVDNA水平、HBeAg的状态以及ALT水平可以预测HBV感染的长期结局如肝硬化或肝细胞癌。本文对HBV感染自然史分期、慢性HBV感染的结局和预后进行了综述。     

乙型肝炎病毒X变异体穿梭载体的成功构建及在酵母中的表达

目的构建表达乙肝病毒（HBV）X变异体基因的穿梭载体。方法用PCR法扩增HBVX基因及变异体基因序列，用EcoRⅠ和PstⅠ双酶切pAS2-1和PCR产物．连接酶切片段pAS2—1及X变异体片段以构建pAS2-1-（X变异体）穿梭质粒，并通过醋酸锂转化法转化入酵母AH109中。结果已构建的pAS2-1-（X变异体）穿梭质粒经序列测定含有所需的HBV—X变异体基因片段，转入酵母细胞后经PCR证实酵母内有表达。结论成功构建了pAS2-1-X变异体穿梭质粒，为进一步在真核细胞内更全面了解HBV-X蛋白作用奠定了基础。     

Contributions of transgenic mouse studies on the research of hepatitis B virus and hepatitis C virus-induced hepatocarcinogenesis

Transgenic mouse technology has enabled the investigation of the pathogenic effects, including those on development, immunological reactions and carcinogenesis, of viral genes directly in living organism in a real-time manner. Although viral hepatocarcinogenesis comprises multiple sequences of pathological events, that is, chronic necroinflammation and the subsequent regeneration of hepatocytes that induces the accumulation of genetic alterations and hepatocellular carcinoma(HCC), the direct action of viral proteins also play significant roles. The pathogenesis of hepatitis B virus X and hepatitis C virus(HCV) core genes has been extensively studied by virtue of their functions as a transactivator and a steatosis inducer, respectively. In particular, the mechanism of steatosis in HCV infection and its possible association with HCC has been well studied using HCV core gene transgenic mouse models. Although transgenic mouse models have remarkable advantages, they are intrinsically accompanied by some drawbacks when used to study human diseases. Therefore, the results obtained from transgenic mouse studies should be carefully interpreted in the context of whether or not they are well associated with human pathogenesis.     

MiR-122 in hepatitis B virus and hepatitis C virus dual infection

Hepatitis B virus (HBV) and hepatitis C virus (HCV) infections are the most common causes of chronic liver diseases and hepatocelluar carcinomas. Over the past few years, the liver-enriched microRNA-122 (miR-122) has been shown to differentially regulate viral replication of HBV and HCV. It is notable that the level of miR-122 is positively and negatively regulated by HCV and HBV, respectively. Consistent with the well-documented phenomenon that miR-122 promotes HCV accumulation, inhibition of miR-122 has been shown as an effective therapy for the treatment of HCV infection in both chimpanzees and humans. On the other hand, miR-122 is also known to block HBV replication, and HBV has recently been shown to inhibit miR-122 expression; such a reciprocal inhibition between miR-122 and HBV suggests an intriguing possibility that miR-122 replacement may represent a potential therapy for treatment of HBV infection. As HBV and HCV have shared transmission routes, dual infection is not an uncommon scenario, which is associated with more advanced liver disease than either HBV or HCV mono-infection. Thus, there is a clear need to further understand the interaction between HBV and HCV and to delineate the role of miR-122 in HBV/HCV dual infection in order to devise effective therapy. This review summarizes the current understanding of HBV/HCV dual infection, focusing on the pathobiological role and therapeutic potential of miR-122.     

HBV/HCV重叠感染的抗病毒治疗

从全球范围看,乙型肝炎病毒(hepatitis B virus,HBV)和丙型肝炎病毒(hepatitis C virus,HCV)重叠感染估计约有700-2000万人口感染.重叠感染和单一HBV或HCV感染比较,更易发展为肝硬化、肝细胞癌甚至肝衰竭的比例也高,HBV和HCV重叠感染可有四种不同的临床模式,即HCV活动...     

In vitro resistance to interferon of hepatitis B virus with precore mutation

AIM: Chronic hepatitis B virus (HBV) infection is predominantly treated with interferon alpha (IFN-α), which results in an efficient reduction of the viral load only in 20-40% of treated patients. Mutations at HBV precore prevail in different clinical status of HBV infection. The roles of precore mutation in the progression of chronic hepatitis and interferon sensitivity are still unknown. The aim of this study was to explore if there was any relationship between HBV precore mutation and sensitivity to interferon in vitro. METHODS: HBV replication-competent recombinant constructs with different patterns of precore mutations were developed. Then the recombinants were transiently transfected into hepatoma cell line (Huh7) by calcium phosphate transfection method. With or without IFN, viral products in culture medium were collected and quantified 3 d after transfection. RESULTS: We obtained 4 recombinant constructs by orientation-cloning 1.2-fold-overlength HBV genome into pUC18 vector via the EcoRI and Hind lll and PCR mediated site-directed mutagenesis method. All the recombinants contained mutations within precore region. Huh7 cells transfected with recombinants secreted HBsAg and HBV particles into the cell culture medium, indicating that all the recombinants were replication-competent. By comparing the amount of HBV DNA in the medium, we found that HBV DNA in medium reflecting HBV replication efficiency was different in different recombinants. Recombinants containing precore mutation had fewer HBV DNAs in culture medium than wild type. This result: showed that recombinants containing precore mutation had lower replication efficiency than wild type. HBV DNA was decreased in pUC18-HBV1.2-WT recombinants after IFN was added while others with precore mutations were not, indicating that HBV harboring precore mutation was less sensitive to IFN in cell culture system. CONCLUSION: These data indicate that HBV harboring precore mutation may be resistant to IFN in vitro.