从组织病理学角度探讨慢性乙型肝炎肝纤维化的发生

目的 探讨窦内皮细胞在肝纤维化发生中的作用,以及肝脏微循环障碍、肝窦毛细血管化与肝纤维化发生的关系. 方法对56例慢性乙型肝炎患者进行肝活组织枪查,将肝组织分成两部分,分别通过HE染色进行光学显微镜观察,以及常规制作电子显微镜标本在透射电子显微镜下进行超微结构观察.结果 56例慢性乙型肝炎患者中,轻度39例,中度10例,重度7例.慢性乙型肝炎患者肝星状细胞的形态类似成纤维细胞,周围有胶原纤维形成.窦内皮细胞与肝星状细胞间有电子致密物沉积.其中53例窦内皮细胞窗孔有不同程度的减少,减小,20例见窦内皮细胞下有连续或不连续的膜样物质形成,15例狄氏腔内有胶原纤维沉积.15例肝功能正常患者有的肝窦腔内有红细胞聚集,严重者有微血栓形成. 结论窦内皮细胞通过与肝星状细胞作用,从而参与肝纤维化的发生.肝脏微循环障碍、肝窦毛细血管化是肝纤维化的早期重要病理改变.     

从组织病理学角度探讨慢性乙型肝炎肝纤维化的发生

目的 探讨窦内皮细胞在肝纤维化发生中的作用,以及肝脏微循环障碍、肝窦毛细血管化与肝纤维化发生的关系. 方法对56例慢性乙型肝炎患者进行肝活组织枪查,将肝组织分成两部分,分别通过HE染色进行光学显微镜观察,以及常规制作电子显微镜标本在透射电子显微镜下进行超微结构观察.结果 56例慢性乙型肝炎患者中,轻度39例,中度10例,重度7例.慢性乙型肝炎患者肝星状细胞的形态类似成纤维细胞,周围有胶原纤维形成.窦内皮细胞与肝星状细胞间有电子致密物沉积.其中53例窦内皮细胞窗孔有不同程度的减少,减小,20例见窦内皮细胞下有连续或不连续的膜样物质形成,15例狄氏腔内有胶原纤维沉积.15例肝功能正常患者有的肝窦腔内有红细胞聚集,严重者有微血栓形成. 结论窦内皮细胞通过与肝星状细胞作用,从而参与肝纤维化的发生.肝脏微循环障碍、肝窦毛细血管化是肝纤维化的早期重要病理改变.     

"免疫耐受期"慢性乙型肝炎病毒感染者临床与肝脏病理学特点

目的 了解免疫耐受期慢性HBV感染者临床及肝脏病理学特征.方法 分析98例"免疫耐受期"慢性HBV患者年龄、性别、血清HBV DAN水平、肝脏炎症活动度及纤维化程度、肝脏HBsAg和HBcAg表达情况.并对不同血清ALT水平的肝脏炎症活动度及纤维化程度进行比较.采用X2检验.结果 98例患者中,＜30岁者83例,占84.7%,≥30岁者15例,占15.3%.有48.0%感染者的母亲HBsAg阳性.所有感染者血清HBVDNA为高水平复制,＞1×107拷贝/mL者占78.5%.仅5例感染者肝脏炎症活动度为Go,占5.1%;其中G1 64例,占65.3%;G2 29例,占29.6%.56例肝脏无明显纤维化(SO),占57.1%;S1 23例,占23.5%;S2 14例,占14.3%;S3 5例,占5.1%;未发现肝硬化.肝组织HBsAg阳性79例,占80.6%;HBeAg阳性80例,占81.6%.血清ALT在正常范围高水平的感染者肝脏纤维化程度明显高于ALT低水平者(X2=8.112 3,P=0.043 7).结论绝大部分"免疫耐受期"慢性HBV感染者肝脏存在轻度炎性反应,部分已出现纤维化;对"免疫耐受期"HBV感染者进行肝脏病理学检查,有利于正确判断病情和确定治疗方案.     

Antiviral effect of Chinese medicine jiaweisinisan in hepatitis B virus transgenic mice

AIM: To study the antiviral effect of Chinese medicine jiaweisinisan (JWSNS) on hepatitis B virus (HBV) infection in transgenic mice (TGM). METHODS: Twenty two 6-8 wk old HBV TGM in the third generation were divided into TGM control group and TGM treated group randomly. The normal control group included ten normal BC 57L/6 mice at the same age. The mice in treated group were administrated with JWSNS at the concentration of 4 g/mL and the dosage of 50 g/kg per d for 30 d, while the mice in TGM control group and normal control group were administrated with normal saline at the same dosage and the same time. Polymerase chain reaction (PCR) was used to assess the contents of HBV DNA in serum of HBV TGM before and after treatments, whereas blot hybridization was utilized to measure the contents of HBV DNA in the liver of both HBV TGM and normal BC 57L/6 mice. RESULTS: The levels of serum HBV DNA in TGM treated group were remarkably decreased after the treatment of JWSNS (7.662±0.78 vs 5.22±3.14, P<0.05), while there was no obvious change after administration of normal saline in TGM control group (7.125±4.26 vs 8.932±5.12, P>0.05). The OD values of HBV DNA in the livers of the mice in TGM treated group were significantly lower than those of TGM control group (0.274±0.096 vs 0.432±0.119,P<0.01). CONCLUSION: JWSNS exerts suppressive effects on HBV DNA in the serum and liver of TGM.     

丙型肝炎病毒非编码区ABC程序酶切分型研究

目的为进一步了解中国是否存在HCV 3b基因及1a、2b和6a基因型感染，建立HCV 5′端非编码区(5′ NCR)不同基因型的基因库。方法分型方法按ABC程序进行，A应用BHH′(BsrBⅠ、HaeⅡ、HinfⅠ)复介内切酶消化5′NCR cDNA，可将不同基因型划分为5组：1a、1b，6a，2a、2b，3a，3b、4a。B应用BstU Ⅰ内切酶鉴别1a、1b。C应用Hae Ⅲ内切酶鉴别2a、2b、3b、4a及6a。电泳检测片段大小。结果(1)la、1b、2a、2b、3a、3b、4a、6a 8种基因型参比品的ABC分型结果表明，该8种基因型获得良好的分型效果。(2)93份HCV RNA阳性患者ABC分型结果表明，1b型感染率占66．67％，2a型18．28％，1b／2b型、3b型及2b型均为3．23％，2a／2b型和1b／2a型各为2．15％，1a型1．08％。结论结果表明应用HCV 5′-NCR ABC分型技术既保证了HCV RNA检测的灵敏度，又能完成1a-6a型中的8种基因型的鉴别。     

New therapeutic opportunities for hepatitis C based on small RNA

Hepatitis C virus (HCV) infection is one of the major causes of chronic liver disease, including cirrhosis and liver cancer and is therefore, the most common indication for liver transplantation. Conventional antiviral drugs such as pegylated interferon-alpha, taken in combination with ribavirin, represent a milestone in the therapy of this disease. However, due to different viral and host factors, clinical success can be achieved only in approximately half of patients, making urgent the requirement of exploiting alternative approaches for HCV therapy. Fortunately, recent advances in the understanding of HCV viral replication and host cell interactions have opened new possibilities for therapeutic intervention. The most recent technologies, such as small interference RNA mediated gene-silencing, anti-sense oligonucleotides (ASO), or viral vector based gene delivery systems, have paved the way to develop novel therapeutic modalities for HCV. In this review, we outline the application of these technologies in the context of HCV therapy. In particular, we will focus on the newly defined role of cellular microRNA (miR-122) in viral replication and discuss its potential for HCV molecular therapy.     

Inhibition of hepatitis B virus DNA replicative intermediate forms by recombinant interferon-γ

AIM: To evaluate the in vitro anti-HBV activity of recombinant human IFN-γ, alone and in combination with lamivudine.METHODS: A recombinant baculovirus-HBV/HepG2 culture system was developed which could support productive HBV infection in vitro. Expression of HBsAg and HBeAg in infected HepG2 culture medium was detected by commercial enzyme immunoassays. HBV DNA replication intermediates were detected in infected cells by Southern hybridization and viral DNA load was determined by dot hybridization.RESULTS: IFN-γ at 0.1 to 5 μg/L efficiently down regulated HBsAg expression in transduced HepG2 cells.At 5 μg/L, IFN-γ also suppressed HBV DNA replication in these cells. While treatment with a combination of lamivudine and IFN-γ showed no additive effect,sequential treatment first with lamivudine and then IFN-γ was found to be promising. In this culture system the best HBV suppression was observed with a pulse of 2 μmol/L lamivudine for two days, followed by 1 μg/L IFN-γ for another four days. Compared to treatment with lamivudine alone, the sequential use of 0.2 μmol/L lamivudine for two days, followed by 5 μg/L IFN-γ for six days showed a 72% reduction in HBV cccDNA pool.CONCLUSION: This in vitro study warrants further evaluation of a combination of IFN-γ and lamivudine,especially in IFN-α non-responder chronic hepatitis B patients. A reduced duration of lamivudine treatment would also restrict the emergence of drug-resistant HBV mutants.     

乙型肝炎病毒三种基因分型方法的比较分析

随着乙型肝炎病毒（HBV）全基因序列测定病毒株的增多，近年人们开始根据生物系统发生学中的基因分类法建立HBV的基因分型技术和标准。对HBV进行基因分型有利于对HBV感染的流行病学、病因学和临床诊断与治疗进行更加深入细致的研究，现有研究表明不同亚型HBV感染的临床病程和对治疗的反应都存在一定的差异。我们分别应用自行建立的实时荧光聚合酶链反应（PCR）法、HBVS基因测序法和聚合酶链反应-限制性片段长度多态性（PCR—RFLP）法对75例慢性乙型肝炎患者的血清基因型进行检测，并对检测结果进行分析比较。     

Current Safety of the Blood Supply in the United States

In common with other developed countries, the United States has placed a great deal of emphasis on blood safety. As a result of careful donor selection and the use of advanced tests, including nucleic acid testing (NAT), the risk of transmission of human immunodeficiency virus and hepatitis C virus has been reduced to about 1 in 1.5 million donations. NAT for hepatitis B virus has not been introduced, but nevertheless the risk is low. Attention recently has been focused on emerging infections. NAT for West Nile virus was implemented within 6 to 8 months of recognition of the need to prevent transfusion transmission of this newly introduced virus. Approximately 1000 potentially infectious donations were identified and removed from the blood supply during the 2003 season. Other emerging infections attracting attention include Chagas' disease, babesiosis, malaria, and variant Creutzfeldt-Jakob disease.     

The distribution of HBV, HCV and HGV among livers with fulminant hepatic failure of different aetiology

Background/Aims: The aim of the study was to assess the impact factor of HCV and HGV in fulminant hepatic failure.Methods: The 5′-untranslated regions of HCV RNA and HGV RNA and a segment of the core antigen sequence of HBV were amplified after extracting the nucleic acids from snap-frozen tissue aliquots from explanted livers of 26 consecutive patients undergoing orthotopic liver transplantation for fulminant hepatic failure preoperatively diagnosed as either autoimmune (n=2), HAV/HBV (n=8), toxic (n=4) or aetiologically unknown (n=12).Results: HCV RNA was detected in five of 26 (19.2%) livers with fulminant hepatic failure. All five HCV RNA-positive livers belonged to the group of non-toxic, non-autoimmune liver failure (n=20), three of them were found in the group of liver failure with unknown aetiology (n=12) and two in the group of HBV-associated liver failure (n=7), making an HCV incidence of 25%, 25%, and 28.6%, in the different groups, respectively. HGV RNA was detected in 10 of 17 (58.8%) explants and in all four groups of fulminant hepatic failure as defined preoperatively. HBV DNA was identified in six livers of 26 patients (23.1%) with fulminant hepatic failure. Neither HCV RNA nor HBV DNA was detected in the livers of patients with toxic or autoimmune fulminant hepatic failure.Conclusions: These results indicate that HBV and HCV, but not HGV, play an aetiologic role in fulminant hepatic failure. HCV-positive cases were concentrated either in the group of otherwise unexplained fulminant hepatic failure or in the group of HBV fulminant hepatic failure. HGV-positive cases, on the other hand, were found within all four preoperatively defined groups, indicating a role as cofactor rather than as single aetiologic agent.