流体动力学法乙型肝炎病毒感染动物模型的建立

目的:建立一种简便、有效、稳定的乙型肝炎病毒(HBV)感染的动物模型,观察该模型动物体内不同时间点各种HBV标志物的表达情况．方法:以流体动力学法尾静脉注射BALB／c 小鼠pcDNA3．1-HBV,1 wk内检测各种HBV 标志物,时间分辨免疫荧光分析法(IFMA)检测血清中HBsAg,HBeAg,抗HBs,抗HBe,抗 HBc,荧光定量PCR法(FQ-PCR)检测血清HBV DNA,免疫组织化学法检测肝组织HBsAg和 HBcAg．结果:成功建立一种急性HBV感染动物模型, 第1天血清中HBsAg表达达高峰,后逐渐降低, HBeAg表达量少,分别在第4、5、7天检测到抗HBc,抗HBe,抗HBs,d1 HBV DNA滴度亦达高峰,后渐下降,免疫组织化学示HBsAg呈胞质内弥漫性分布,HBcAg亦主要为胞质型分布．结论:以流体动力学法建立的HBV模型是一种新型有效的HBV感染动物模型,能稳定较高水平表达大部分HBV标志物．     

急性乙型肝炎病毒感染小鼠模型的建立

目的采用尾静脉液压法建立小鼠急性HBV感染的动物模型。方法以液压法将具有复制能力的HBV质粒pAAV-HBV1.2通过尾静脉注射到免疫功能正常的BALB/c小鼠体内,注射后第1、2、4、6、8d,分别采用改良赖氏法、时间分辨免疫荧光法、实时荧光定量PCR检测小鼠血清中谷丙转氨酶(ALT)、HBsAg、HBeAg、抗HBs、抗HBe、HBV DNA的水平,免疫组化检测肝组织HBsAg、HBcAg的表达。结果16只小鼠注射pAAV-HBV1.2后,有14只(85.7%)小鼠在注射后第1d血清中可检测到HBsAg,小鼠血清中HBsAg和HBeAg水平在第1d达高峰,之后逐渐下降,第8d均未能检测到。小鼠血清中HBV DNA在第2d达高峰,之后仍维持在较高水平,至第8d时为1.9×104copies/mL。至第8d肝组织中可见约5%的HBcAg阳性肝细胞和2%的HBsAg阳性肝细胞。结论采用尾静脉液压法成功的建立了小鼠急性HBV感染的动物模型。     

建立幽门螺杆菌感染BALB/c小鼠动物模型的实验研究

[目的]建立幽门螺杆菌(Hp)BALB/c小鼠感染动物模型。[方法]用cagA基因和vacA基因同时阳性的Hp小鼠适应株给BALB/c小鼠灌胃。[结果]经Hp小鼠适应株灌胃4周后,病理模型组10只感染的小鼠胃黏膜尿素酶试验、细菌学培养和组织病理学检查均为阳性,感染率为100%;正常对照组未见Hp生长,两者比较差异有统计学意义(P<0.05)。[结论]成功地建立了云南Hp小鼠适应株感染的BALB/c小鼠动物模型。这种ca gA基因和vacA基因同时阳性的Hp小鼠适应株毒力强容易定植;可在普通饲养条件下建立,即有利应用于云南地区Hp的相关研究,又可节约研究经费,易于推广使用。     

HBV小鼠动物模型的研发进展

HBV是导致肝硬化和肝细胞癌的主要原因。目前全球约有2.4亿HBV感染者,其中每年约100万人死于HBV所引起的肝脏疾病,是全球主要的公共卫生问题之一。然而HBV是宿主种属特异性极强的嗜肝病毒,只能在几种特定的动物中感染复制。因此,研究支持HBV感染的动物模型,对HBV感染和致病机制的认识,以及有效治疗方案的快速研发具有重要的作用。当前,HBV感染模型的不断升级有力推动了HBV的病毒学研究,特别是HBV小鼠模型的不断开发为HBV研究提供了便捷的研究工具,本文将对这一领域的研究作一综述。     

广西麻鸭乙型肝炎感染病毒动物模型的实验研究

目的探讨广西麻鸭作为乙型肝炎病毒感染动物模型的可能性。方法应用广西1 d龄雏麻鸭经腹腔感染鸭乙型肝炎病毒(DHBV),13 d后采用实时荧光定量PCR法检测麻鸭血清DHBV含量,筛选出DHBV强阳性鸭。结果共检测148份麻鸭血清标本,其中DHBV阳性标本136份,阳性率为91.9%。结论广西麻鸭可作为乙型肝炎病毒感染动物模型。实时荧光定量PCR法检测DHBV敏感性较高,重复性好,可用于DHBV检测。     

鸭乙型肝炎病毒感染模型的研究进展

HBV和DHBV均为嗜肝DNA病毒,它们都有相似的基因组和病毒结构特征,以及通过RNA逆转录复制的特性。DHBV感染鸭模型是研究HBV基因突变、细胞表面病毒受体、病毒清除机制以及筛选新的抗病毒药物的合适的动物模型。文章就鸭乙型肝炎病毒感染模型的研究情况作一简要综述。     

小分子肽抗乙型肝炎病毒的实验研究

目的:应用2.2.15细胞株和鸭乙型肝炎病毒(DHBV)感染模型,评价小分子肽CMS 017抗乙肝病毒作用.方法:①CMS 017以倍比稀释浓度加入2.2.15细胞培养中,作用8天后吸取培养上清,PCR法检测HBVDNA.②建立DHBV感染模型,CMS017以60、200、600μg/kg·d-13个剂量腹腔注射给药28天,检测用药前、后血清DHBV DNA、DHBsAg变化.结果:①CMS 017体外抑制DHBV呈量效关系,IC50为2.3μg/ml.②鸭体内实验中,CMS 017中、高剂量组用药7天、14天开始出现血清DHBV DNA降低(P＜0.01,P＜0.05),持续至停药后7天.CMS 017中剂量组用药后血清DHBsAg降低的时效关系与DHBV DNA完全一致.结论:CMS017具有抗乙肝病毒作用,其体内作用的量效关系待确定.     

树Qu实验感染乙型肝炎病毒的再评价

目的 评价乙肝病毒（ＨＢＶ）实验感染树Ｑｕ的稳定性与实用性。方法 用含ＨＢＶ的２．２．１５细胞或乙肝患者混合血清以不同剂量，经不同途径接种６０只健康树Ｑｕ，观察其临床表现，检测血清ＡＬＴ、ＨＢｓＡｇ、ＨＢｅＡｇ及ＨＢＶ－ＤＮＡ，肝组织行ＨＥ及免疫组织化学染色。结果 部分树Ｑｕ出现一过性食欲不振、稀便；６０只树Ｑｕ不同时粗ＡＬＴ水平均在正常范围；８只树ＱｕＨＢＶ标志物一过性阳性；肝组织未见类似于人乙     

919糖浆抗鸭乙型肝炎病毒的实验研究

目的:研究919糖浆时鸭乙型肝炎的体内抗病毒作用。方法:以感染鸭乙型肝炎病毒(DHBV)的武汉麻鸭为实验动物模型。使1d龄武汉麻鸭感染DHBV,7d后用Dot-EIA法筛选出DHBsAg强阳性鸭。随机分组后给予919糖浆治疗14d,于感染后第7d(为用药前)、用药7d、14d及停药后3d分别采血,采用斑点杂交方法检测血清DHBVDNA。结果。用药919糖浆7d、14d鸭血清DHBV DNA OD值均明显低于给药前,有显著性差异(P<0.05,P<0.01)。停药3d后没有反跳现象。结论:919糖浆有一定的抑制DHBV DNA复制作用。     

HBV感染者HCV的重叠感染关系研究

目的 研究HBV感染患者中HCV的重叠感染状况及其相互关系。 方法 采用ELISA法对767例HBV感染患者同步检测HBV和HCV血清标志物,对可疑HCV感染但抗HCV阴性和/或抗-HCV阳性患者血清,采用PCR法检测HCV-RNA。 结果 HCV重叠感染率为4.82％,且在各类乙肝患者中存在非常显著差异(P<0.01);HBV/HCV感染组重症肝炎的发生率显著高于非HCV感染组(P<0.01);HBV/HCV感染组HBsAg阳性率显著低于单纯HBV感染组(P<0.05);HBV/HCV感染组HCV-RNA阳性率显著低于单纯HCV感染组(P<0.05)。 结论 HCV重叠感染与乙肝患者的发病、病情加重及重症肝炎的发生相关;HCV可抑制或中止HBsAg携带状态,但这种作用远不如对病情的加重作用重要;同时HBV对HCV的复制亦存在抑制作用。     

HBsAg-negative hepatitis B virus infections in hepatitis C virus-associated hepatocellular carcinoma

This study was conducted to evaluate reports that hepatitis B virus (HBV) DNA sequences can be found in the serum and/or tumour tissue from some hepatocellular carcinoma (HCC) patients who have no detectable hepatitis B surface antigen (HBsAg) in their sera. Such HBV infections would be highly atypical, because prospective studies have shown a clear succession of specific serologic markers during and after most HBV infections. As most HBsAg-negative HCC patients in Japan have hepatitis C virus (HCV) infections, the present study was conducted to determine whether some of these patients actually have unrecognized HBV infections. Thirty newly diagnosed HCC patients from Kurume, Japan, with antibody to the hepatitis C virus (anti-HCV) were studied. None of the 30 had HBsAg detectable in their serum. Of 22 for whom test results for antibodies to the hepatitis B core antigen (anti-HBc) and antibodies to HBsAg (anti-HBs) were available, 14 (64%) had anti-HBc and anti-HBs, four (18%) had anti-HBc alone, and four (18%) had no HBV markers. Nested polymerase chain reaction was used to detect the HBV surface (S), core (C), polymerase (P) and core promoter gene sequences in the HCC tissues and in the adjacent nontumorous liver tissues. HBV DNA was detected in HCC and/or adjacent nontumorous liver in 22 of 30 (73%) patients [detected in both HCC and nontumorous liver in 19/30 patients (63%)]. Among the 22 patients with detectable HBV DNA, more than one HBV gene was detected in 10 (46%). Among the four patients whose sera were negative for all HBV markers, three had HBV DNA in either HCC and nontumorous liver (two cases) or only in the nontumorous liver (one case); HBV DNA could not be detected in tissues from the fourth patient. In 18 of 21 (86%) patients with detectable HBV core promoter sequences, mutations at both nucleotides 1762 (A-GT) and 1764 (G-A) in the core promoter region were found. No deletions were detected in the core promoter gene region of the type reported to be associated with some cases of HBsAg-negative HBV infection. Thus, HBV DNA was detectable in 22 (73%) HBsAg-negative, anti-HCV-positive HCCs, including three (10%) who were also negative for anti-HBc and anti-HBs. HBV mutations at both nucleotides 1762 (A-GT) and 1764 (G-A) in the core promoter region were found in the majority of cases, mutations that have previously been reported in HBV that is integrated in HCC DNA. In serologic surveys to determine etiologic associations of HCC, patients such as those in this study would have been incorrectly designated as having 'HCV-associated HCC,' whereas the data in this study suggest that HBV could have played a role in the development of their HCCs.     

HBV感染小鼠模型的研究进展

HBV由于其较强的种属特异性,目前尚缺乏理想的实验动物模型来阐明其具体的发病机制。目前有关HBV感染模型的研究大多集中在小鼠方面,并已取得了很大进展。综述了HBV转基因小鼠、HBV转染小鼠和人鼠嵌合肝脏HBV小鼠等模型的优缺点,提出合理使用这些模型有助于更好地阐明HBV的致病机制。     

HIV/hepatitis C virus and HIV/hepatitis B virus coinfections protect against antiretroviral-related hyperlipidaemia

Introduction

Hyperlipidaemia is a recognized complication of HIV antiretroviral therapy. The interactions among HIV, viral hepatitis, antiretroviral therapies and lipids are poorly understood.

Methods

Ontario HIV Treatment Network Cohort Study participants with at least one lipid level after highly active antiretroviral therapy (HAART) initiation were assessed. Hepatitis B virus (HBV)‐ and hepatitis C virus (HCV)‐coinfected patients were identified by serology or chart review. HCV antiviral recipients, diabetics and those on lipid‐lowering drugs at baseline were excluded from the study. Factors associated with a decreased risk of grade 3 or 4 hyperlipidaemia or lipid‐lowering drug use were assessed by multivariate logistic regression.

Results

A total of 1587 HIV‐monoinfected, 190 HIV/HBV‐coinfected and 255 HIV/HCV‐coinfected patients were evaluated. Most were male (85–92% for the 3 groups evaluated: HIV, HIV/HBV, HIV/HCV). The median [interquartile range (IQR)] age at HAART initiation was 48 (44–56) years and was similar between groups. The median (IQR) CD4 count at HAART initiation was 245 (120–370) cells/μL in HIV‐monoinfected participants, 195 (110–330) cells/μL in HIV/HBV‐coinfected participants and 268 (140–409) cells/μL in HIV/HCV‐coinfected participants. Factors associated with a decreased risk of grade 3 or 4 hyperlipidaemia or lipid‐lowering drug use included HIV/HCV coinfection [odds ratio (OR) 0.46; 95% confidence interval (CI) 0.34, 0.61; P<0.0001], HIV/HBV coinfection (OR 0.74; 95% CI 0.55, 0.99; P=0.04), year of starting HAART after 2004 vs. 1997 or earlier (OR 0.37; 95% CI 0.29, 0.48; P<0.0001) and year of starting HAART between 1998 and 2003 vs. 1997 or earlier (OR 0.75; 95% CI 0.61, 0.92; P<0.01). Factors associated with increased risk included age (OR 1.55; 95% CI 1.39, 1.72; per 10 years, P<0.0001) and male gender (OR 1.84; 95% CI 1.36, 2.48; P<0.0001).

Conclusions

HIV/HCV and to a lesser extent HIV/HBV coinfections are protective against HAART‐related hyperlipidaemia.     

Vietnamese community screening for hepatitis B virus and hepatitis C virus

Summary. Asian Americans represent an important cohort at high risk for viral hepatitis. To determine the prevalence of Hepatitis B virus (HBV) and Hepatitis C virus (HCV) infection and HBV vaccination in a Vietnamese community, a total of 322 Vietnamese subjects from a local doctor’s office and annual Vietnamese Health Fair were included in this study. Demographic and clinical data were collected. 2.2% of the screened cohort tested positive for anti‐HCV and 9.3% tested positive for HBsAg. Unlike HBV‐positive subjects, HCV‐positive subjects had significantly higher liver enzymes (P = 0.0045 and P = 0.0332, respectively). The HBV‐positive group was more likely to report jaundice (P = 0.0138) and a family history of HBV (P = 0.0115) compared to HBV‐negative subjects. Forty‐eight patients (15.5%) reported a family history of liver disease (HBV, HCV, HCC, cirrhosis, other). Of this 48, 68.8% reported no personal history of HBV vaccination and 77.1% reported no family history of vaccination for HBV. Among the 183 subjects without a family history of liver disease, 156 (85.2%) reported no personal history of vaccination and 168 (91.8%) reported no family history of vaccination. HBV vaccination rates in those reporting a family history of liver disease were significantly higher (P = 0.020). There was a high prevalence of HBV infection in this community screening. Nevertheless, the rate for HBV vaccination was low. The low prevalence of abnormal liver enzymes in HBV‐positive subjects emphasizes the need for screening to be triggered by risk factors and not by abnormal liver enzymes.     

Japanese case of hepatitis B virus genotypes C/D hybrid

Hepatitis B virus (HBV) is classified into eight genotypes based on complete genome sequence. Each genotype is related to geographic distribution and race. In Japan, most of the genotypes are B and C. In the present study, we report the first Japanese strain of HBV having a recombination between genotypes C and D. A 30-year-old woman was admitted to Kobe Medical Center because of liver dysfunction. She was diagnosed with spontaneous reactivation of chronic hepatitis B. She had no history of blood transfusion and her parents were negative for HBV. The phylogenetic analysis based on the complete genome sequences revealed that this strain was classified into genotype C, whereas the analysis based on Sgene sequence showed that this strain was genotype D. By using a SimPlot program, this strain was confirmed as a recombinant strain between genotypes C and D. Compared with previous recombinant strains in China, the breakpoint was the same and the difference was only 0.8% of the complete genome sequence. It was unclear whether or not this strain was transmitted from China, but the recombinant strains and intergenotypes of HBV have already existed in Japan.     

De novo hepatitis B virus infection in hepatocellular carcinoma following eradication of hepatitis C virus by interferon therapy

Epidemiological studies have revealed that hepatocellular carcinoma (HCC) is still observed in hepatitis C virus (HCV)‐positive patients with a sustained response to interferon (IFN) treatment, although a substantial decrease in the incidence of hepatocellular carcinoma (HCC) has been achieved in those patients. Why HCC develops in patients who have a complete clearance of HCV remains unclear. Here, we provided evidence of latent hepatitis B virus (HBV) infection in an initially HCV‐positive chronic hepatitis patient who developed HCC after the complete eradication of HCV by IFN therapy. Although he was initially negative for anti‐hepatitis B surface antigen (HBsAg) or circulating HBV DNA but positive for anti‐hepatitis B core antigen (anti‐HBc) in his sera, he developed HBsAg and HBV DNA during the course of the management of a series of cancers. HBV DNA was detectable in the liver tissues before HBV reactivation and the viral sequences derived from his anti‐HBc‐positive liver showed 100% homology to that from the serum after HBsAg appearance. These findings indicates that HCV‐positive individuals who are positive for anti‐HBc in the absence of HBsAg could have latent HBV infection in their liver tissues and intrahepatic HBV infection may play a pivotal role in the development of HCC after the IFN‐mediated eradication of HCV.