小儿感染庚型肝炎病毒后肝脏病毒核酸分布与临床关系的研究

目的 探讨小儿庚型肝炎病毒感染、肝脏庚型肝炎病毒核糖核酸 (HGVRNA)分布与其他HGV标志、肝脏病理损害及与临床的关系。方法 应用原位杂交 (ISH)检测 17例小儿乙型肝炎患儿肝脏HGVRNA。结果 17例血清抗 HGV阳性者 ,其中肝脏HGVRNA阳性 9例 (9/ 17,5 3% )。肝脏HGVRNA分布在汇管区周围 ,在肝小叶呈散在分布 ,多呈胞浆型 ;肝脏HGVRNA阳性组病变和阴性组比较以轻度 (G2 级以下 )为多 ,与肝纤维化无关 ;与肝脏HGAg无相关性 ,与抗 HGV相关(P =0 .0 13) ;与血清丙氨酸转氨酶 (ALT)、血清胆红素 (SB)和蛋白电泳的γ球蛋白升高无相关性。结论HGV即使有一定的致病性 ,其致病力也很弱。对HGV的致病性还需做进一步深入研究。

《肝脏靶向腺相关病毒(AAV)血友病B基因治疗研究》解读

本文最初发表在2022年《Lancet Haematology》杂志上,发表后引起业内广泛关注,宾夕法尼亚大学Lindsey A George教授在《Lancet Haematology》杂志同期给予高度评价,《Molecular Therapy》杂志也对该研究进行了重点介绍(Molecular Therapy, 2022,30(7):2394-2396)。本文文章题录为:Feng Xue, Huiyuan Li, Xia Wu, et al.Safety and activity of an engineered, liver-tropic adeno-associated virus vector expressing a hyperactive Padua factorⅨadministered with prophylactic glucocorticoids in patients with haemophilia B:a single-centre, single-arm, phase 1,pilot trial[J].Lancet Haematology.2022,9(7):e504-e513。该研究首次证实预防性使用糖皮质激素的肝脏靶向腺相关病毒(AAV)血友病B基因治疗能够安全、快速、稳定地提高FⅨ活性,并探索性应用单细胞技术发现糖皮质激素能够减轻基因治疗带来的免疫反应,为中国血友病B基因治疗新策略奠定了理论和实践基础。经通讯作者许可,再次通过佳文解读的方式来阐述这一研究。

观察血小板淋巴细胞比(PLR)、中性粒细胞淋巴细胞比(NLR)和淋巴细胞单核细胞比(LMR)在乙型肝炎病毒相关肝脏疾病中的变化及其临床意义

目的分析血小板淋巴细胞比(PLR)、中性粒细胞淋巴细胞比(NLR)和淋巴细胞单核细胞比(LMR)在乙型肝炎病毒相关肝脏疾病中的变化及其临床意义。方法在2018年1月至2018年12月到我院进行治疗的HBV感染孕产妇中,选取240例作为本次的研究对象,将其分为HBV感染孕妇组、乙肝活动期孕妇组和乙肝活动期产妇组,各80例,选取同一时期到我院进行产检的健康孕产妇60例作为本次研究的对照组。对比各组受检者的PLR、NLR、LMR指标水平。结果HBV感染孕妇组、乙肝活动期孕妇组的PLR无明显差异(P>0.05),且均低于对照组(P<0.05),乙肝活动期产妇组的PLR高于对照组(P<0.05);HBV感染孕妇组的NLR与对照组无明差异(P>0.05),3组HBV相关疾病组之间的NLR具有明显差异(P<0.05),且HBV感染孕妇组<乙肝活动期孕妇组<乙肝活动期产妇组;在LMR方面,乙肝活动期产妇组<乙肝活动期孕妇组<HBV感染孕妇组<对照组(P<0.05)。结论PLR、NLR、LMR在乙型肝炎病毒相关肝脏疾病的进展中具有一定的变化规律,其中LMR对正常健康孕产妇、HBV感染孕产妇和乙肝活动期孕产妇具有良好的鉴别诊断价值,三者存在各自的特点和优势,为乙型肝炎病毒相关肝脏疾病的临床诊断提供了重要指导。

丙硫异烟胺治疗老年复治肺结核的肝脏毒性及应对措施

目的探讨丙硫异烟胺治疗老年复治肺结核对肝脏的毒性及应对措施。方法本次研究选取我院内科于2013年9月至2014年11月医治的120例老年复治肺结核患者,随机分为观察组与对照组两组,采用抗结核药物及保肝药物,观察其肝脏毒性及应对措施。结果 2组肝功能出现异常时间无统计学意义(P>0.05);观察组肝功能异常恢复时间短于对照组,疗效可观,具有统计价值(P<0.05)。结论肺结核老年患者采用丙硫异烟胺治疗过程中出现肝功能异常,对其短时间停用药物服用保肝类药物,处理效果可观,且利于保护患者肝功能,不影响患者痰菌转阴等情况。

Assessment of correlation between serum titers of hepatitis c virus and severity of liver disease

AIM:The significance of hepatitis C virus (HCV) serum titers has been examined in several clinical situations. There is much evidence that patients with a lower viral load have better response rates to anti-viral therapy compared to those with higher levels.Moreover,a direct association has been observed between serum titers of HCV and transmission rates of the virus.The aim of the present study was to determine if there was any correlation between HCV viral load and the severity of liver disease. METHODS:Fifty patients with HCV infection were included in the study.These comprised of 34 subjects with a history of alcohol use and 16 non-alcoholics.Quantitative serum HCV RNA assay was carried out using the branched DNA (bDNA) technique.Linear regression analysis was performed between serum viral titers and liver tests.In addition,for the purpose of comparison,the subjects were divided into two groups:those with low viral liters (≤50 genome mEq/mL) and high titers (>50 mEq/mL). RESULTS:All subjects were men,with a mean±SD age of 47±7.8 years.The mean HCV RNA level in the blood was 76.3×10~5±109.1 genome equivalents/mL.There was no correlation between HCV RNA levels and age of the patients (r=0.181),and the history or amount (g/d) of alcohol consumption (r=0.07).Furthermore,no correlation was observed between serum HCV RNA levels and the severity of liver disease as judged by the values of serum albumin (r=0.175),bilirubin (r=0.217),ALT (r=0.06) and AST (r=0.004) levels.Similarly,no significant difference was observed between patients with low viral titers and high liters with respect to any of the parameters. CONCLUSION:Our results indicate that the severity of liver disease is independent of serum levels of hepatitis C virus.These findings are important since they have a direct impact on the current debate regarding the role of direct cytopathic effect of hepatitis C virus versus immune-mediated injury in the pathogenesis of HCV-related liver damage.

Fibrinogen-like protein 2 fibroleukin expression and its correlation with disease progression in murine hepatitis virus type 3-induced fulminant hepatitis and in patients with severe viral hepatitis B

AIM： To evaluate the expression of fibrinogenlike protein 2 （fgl2） and its correlation with disease progression in both mice and patients with severe viral hepatitis. METHODS： Balb/cJ or A/J mice were infected intraperitoneally （ip） with 100 PFU of murine hepatitis virus type 3 （MHV-3）, liver and serum were harvested at 24, 48, and 72 h post infection for further use. Liver tissues were obtained from 23 patients with severe acute chronic （AOC） hepatitis B and 13 patients with mild chronic hepatitis B. Fourteen patients with mild chronic hepatitis B with cirrhosis and 4 liver donors served as normal controls. In addition, peripheral blood mononuciear cells （PBMC） were isolated from 30 patients （unpaired） with severe AOC hepatitis B and 10 healthy volunteers as controls. Procoagulant activity representing functional prothrombinase activity in PBMC and white blood cells was also assayed. A polyclonal antibody against fgl2 was used to detect the expression of both mouse and human fgl2 protein in liver samples as well as in PBMC by immunohistochemistry staining in a separate set of studies. Alanine aminotransferase （ALT） and total bilirubin （TBil） in serum were measured to assess the severity of liver injury.RESULTS： Histological changes were found in liver sections 12-24 h post MHV-3 infection in Balb/cJ mice. In association with changes in liver histology, marked elevations in serum ALT and TBil were observed. House fgl2 （mfgl2） protein was detected in the endothelium of intrahepatic veins and hepatic sinusoids within the liver 24 h after MHV-3 infection. Liver tissues from the patients with severe AOC hepatitis B had classical pathological features of acute necroinflammation. Human fgl2 （hfgl2） was detected in 21 of 23 patients （91.30%） with severe AOC hepatitis B, while only 1 of 13 patients （7.69%） with mild chronic hepatitis B and cirrhosis had hfgl2 mRNA or protein expression. Twenty-eight of thirty patients （93.33%） with severe AOC hepatitis B and 1 of 10 with mild chronic hepatitis B had detectable hfgl2 expression in PBMC. No hfgl2 expression was found either in the liver tissue or in the PBMC from normal donors. There was a positive correlation between hfgl2 expression and the severity of the liver disease as indicated by the levels of TBil. PCA significantly increased in PBMC in patients with severe AOC hepatitis B. CONCLUSION： The molecular and cellular results reported here in both mice and patients with severe viral hepatitis suggest that virus-induced hfgl2 prothrombinase/fibroleukin expression and the coagulation activity associated with the encoded fgl2 protein play a pivotal role in initiating severe hepatitis. The measurement of hfgl2/fibroleukin expression in PBMC may serve as a useful marker to monitor the severity of AOC hepatitis B and a target for therapeutic intervention.

Epidemiology of Hepatitis B and Associated Liver Diseases in China

Hepatitis B virus (HBV) infection has long been a critical public health challenge in China. National surveys revealed a prevalence of approximate 10% for chronic HBV infection in general population. HBV has been the leading cause of chronic hepatitis, cirrhosis, and liver cancers in Chinese population and a common pathogen of acute viral hepatitis. Meanwhile, the epidemic provided important opportunities to research the natural history, public health impact, and therapeutic and preventive interventions for HBV in China. In this review, we summarized the selected key epidemiological studies since 1970s regarding HBV infection and its associated liver diseases in China, and provided considerations for future research, prevention and treatment of HBV.

Initial steroid-free immunosuppression after liver transplantation in recipients with hepatitis c virus related cirrhosis

AIM:Steroids can increase hepatitis C virus(HCV) replication.After liver transplantation(LTx),steroids are commonly used for immunosuppression and acute rejection is usually treated by high steroid dosages.Steroids can worsen the outcome of recurrent HCV infection.Therefore, we evaluated the outcome of HCV infected liver recipients receiving initial steroid-free immunosuppression. METHODS:Thirty patients undergoing LTx received initial steroid-free immunosuppression.Indication for LTx included 7 patients with HCV related cirrhosis.Initial immunosuppression consisted of tacrolimus 2×0.05mg/kg.d po and mycophenolate mofetil(MMF)2×15mg/kg.d po.The tacrolimus dosage was adjusted to trough levels in the target range of 10-15μg/L during the first 3 mo and 5-10μg/L thereafter.Manifestations of acute rejection were verified histologically. RESULTS:Patient and graft survival of 30 patients receiving initial steroid-free immunosuppression was 86% and 83% at 1 and 2 years.Acute rejection occurred in 8/30 patients, including 1 HCV infected recipient.All HCV-infected patients had HCV genotype Ⅱ(lb).HCV seropositivity occurred within the first 4 mo after LTx.The virus load was not remarkably increased during the first year after LTx.Histologically,grafts had no severe recurrent hepatitis. CONCLUSION:From our experience,initial steroid-free immunosuppression does not increase the risk of acute rejection in HCV infected liver recipients.Furthermore,none of the HCV infected patients developed serious chronic liver diseases.It suggests that it may be beneficial to avoid steroids in this particular group of patients after LTx.

Animal models for studying hepatitis C and alcohol effects on liver

Chronic consumption of ethanol has a dramatic effect on the clinical outcome of patients with hepatitis C virus (HCV) infection, but the mechanism linking these two pathologies is unknown. Presently, in vitro systems are limited in their ability to study the interaction between a productive wild-type HCV infection and chronic ethanol exposure. Mouse models are potentially very useful in dissecting elements of the HCV-ethanol relationship. Experiments in mice that transgenically express HCV proteins are outlined, as are experiments for the generation of mice with chimeric human livers. The latter models appear to have the most promise for accurately modeling the effects of chronic ethanol intake in HCV-infected human livers.

Prevalence of porcine endogenous retrovirus in Chinese pig breeds and in patients treated with a porcine liver cell-based bioreactor

AIM： To determine the prevalence of porcine endogenous retrovirus （PERV） in various pig breeds raised in China including Chinese experimental mini-pigs by PERV-reverse transcriptase （PERV-RT enzyme）. Moreover, the potential for infection of PERV was investigated in patients treated with a bioreactor based on porcine liver cells （n = 3）. METHODS： Pig serum, liver and muscle cell-free supernatants were collected from various Chinese pig breeds. Porcine hepatocytes were isolated with a two-step perfusion method. Three patients with acute or chronic liver failure were treated with a bioartificial liver support system （BALSS） for 8-12 h and serum samples were collected from the patients before, immediately after and 30 d after treatment. RESULTS： The activities of PERV-RT enzyme in pig liver and muscle cell-free supernatants were higher than in normal human controls. PERV-TR enzyme activity did not increase in patients before and after 1 mo of treatment. PERV-RT activities were not significantly different when compared with pre-treatment group （1.544±0.155576）, the post-treatment groups （1.501±0.053507, 1.461±0.033808 and 1.6006667±0.01963 for 0, 14 and 30 d post-treatment, respectively, P〉0.05）, and normal control group （1.440±1.0641, P〉0.05）. RT enzyme activity in Chinese experimental mini-pigs was higher than in normal human control group （1.440±1.0641 U/mL, P〈0.05）, and not significantly different （P〉0.05） when compared with the pig breeds except in the muscle supernatants. All the samples including muscle and liver cell supernatants from the Chinese mini-experimental pigs and the four domestic Chinese pig breeds contained PERVs. CONCLUSION： These results suggest that the risk of PERV infection through BALSS containing porcine liver cells without immunosuppressants may be quite low. Although there were PERVs in Chinese experimental mini-pigs and porcine liver cell culture suspensions, we did not find any evidence of persistent PERV infection in patients treated with this porcine hepatocyte-based bioartificial liver.

Hepatitis B and C infection and liver disease trends among human immunodeficiency virus-infected individuals

AIM:To examine trends in and correlates of liver disease and viral hepatitis in an human immunodeficiency virus (HIV)-infected cohort. METHODS:The multi-site adult/adolescent spectrum of HIV-related diseases (ASD) followed 29 490 HIVinfected individuals receiving medical care in 11 U.S. metropolitan areas for an average of 2.4 years,and a total of 69 487 person-years,between 1998 and 2004. ASD collected data on the presentation,treatment,and outcomes of HIV,including liver disease,hepatitis screening,and hepatitis diagnoses. RESULTS:Incident liver disease,chronic hepatitis B virus (HBV),and hepatitis C virus (HCV) were diagnosed in 0.9,1.8,and 4.7 per 100 person-years. HBV and HCV screening increased from fewer than 20% to over 60% during this period of observation (P < 0.001). Deaths occurred in 57% of those diagnosed with liver disease relative to 15% overall (P < 0.001). Overall 10% of deaths occurred among individuals with a diagnosis of liver disease. Despite care guidelines promoting screening and vaccination for HBV and screening for HCV,screening and vaccination were not universally conducted or,if conducted,not documented. CONCLUSION:Due to high rates of incident liver disease,viral hepatitis screening,vaccination,and treatment among HIV-infected individuals should be a priority.

Mechanisms of alcohol-mediated hepatotoxicity in human-immunodeficiency-virus-infected patients

Clinical observations have demonstrated that excessive chronic alcohol use negatively affects human immuno- deficiency virus (HIV) infection and contributes to the liver manifestations of the disease, even in HIV mono- infection. HIV/hepatitis C virus (HCV) co-infection is as- sociated with increased progression of HVC liver disease compared to HCV infection alone, and both of these are negatively affected by alcohol use. Recent data suggest that alcohol use and HIV infection have common targets that contribute to progression of liver disease. Both HIV infection and chronic alcohol use are associated with increased gut permeability and elevated plasma levels of lipopolysaccharide; a central activator of inflammatory responses. Both alcoholic liver disease and HIV infec tionresult in non-specific activation of innate immunity, proinflammatory cytokine cascade upregulation, as well as impaired antigen presenting cell and dendritic cell functions. Finally, alcohol, HIV and antiretroviral therapyaffect hepatocyte functions, which contributes to liver damage. The common targets of alcohol and HIV infection in liver disease are discussed in this minireview.

Biochemical mechanisms in drug-induced liver injury:Certainties and doubts

Drug-induced liver injury is a significant and still unresolved clinical problem. Limitations to knowledge about the mechanisms of toxicity render incomplete the detection of hepatotoxic potential during preclinical development. Several xenobiotics are lipophilic substances and their transformation into hydrophilic compounds by the cytochrome P-450 system results in production of toxic metabolites. Aging, preexisting liver disease, enzyme induction or inhibition, genetic variances, local 02 supply and, above all, the intrinsic molecular properties of the drug may affect this process. Necrotic death follows antioxidant consumption and oxidation of intracellular proteins, which determine increased permeability of mitochondrial membranes, loss of potential, decreased ATP synthesis, inhibition of Ca^2＋-dependent ATPase, reduced capability to sequester Ca^2＋ within mitochondria, and membrane bleb formation. Conversely, activation of nucleases and energetic participation of mitochondria are the main intracellular mechanisms that lead to apoptosis. Non-parenchymal hepatic cells are inducers of hepatocellular injury and targets for damage. Activation of the immune system promotes idiosyncratic reactions that result in hepatic necrosis or cholestasis, in which different HLA genotypes might play a major role. This review focuses on current knowledge of the mechanisms of drug-induced liver injury and recent advances on newly discovered mechanisms of liver damage. Future perspectives including new frontiers for research are discussed.

New perspectives in occult hepatitis C virus infection

Occult hepatitis C virus (HCV) infection, defined as the presence of HCV RNA in liver and in peripheral blood mononuclear cells (PBMCs) in the absence of detectable viral RNA in serum by standard assays, can be found in anti-HCV positive patients with normal serum levels of liver enzymes and in anti-HCV negative patients with persistently elevated liver enzymes of unknown etiology. Occult HCV infection is distributed worldwide and all HCV genotypes seem to be involved in this infection. Occult hepatitis C has been found not only in anti-HCV positive subjects with normal values of liver enzymes or in chronic hepatitis of unknown origin but also in several groups at risk for HCV infection such as hemodialysis patients or family members of patients with occult HCV. This occult infection has been reported also in healthy populations without evidence of liver disease. Occult HCV infection seems to be less aggressive than chronic hepatitis C although patients affected by occult HCV may develop liver cirrhosis and even hepatocellular carcinoma. Thus, anti-HCV negative patients with occult HCV may benefit from antiviral therapy with pegylated-interferon plus ribavirin. The persistence of very low levels of HCV RNA in serum and in PBMCs, along with the maintenance of specific T-cell responses against HCV-antigens observed during a long-term follow-up of patients with occult hepatitis C, indicate that occult HCV is a persistent infection that is not spontaneously eradicated. This is an updated report on diagnosis, epidemiology and clinical implications of occult HCV with special emphasis on anti-HCV negative cases.

Hepatoxicity associated with weight-loss supplements: A case for better post-marketing surveillance

There is a growing number of case reports of hepatoxicity from the widely marketed weight-loss supplement Hydroxycut, which contains the botanical ingredient Garcinia cambogia . These case reports may substantially undercount the true magnitude of harm. Based on the past experience with harmful dietary supplements, US regulators should assume the more precautionary approach favored by Canada and Europe. Lacking effective adverse event surveillance for supplements, or the requirements to prove safety prior to coming to the market, case reports such as those summarized here assume added importance.

Development of Novel Antiviral Therapies for Hepatitis C Virus

Over 170 million people worldwide are infected with hepatitis C virus (HCV), a major cause of liver diseases. Current interferon-based therapy is of limited efficacy and has significant side effects and more effective and better tolerated therapies are urgently needed. HCV is a positive, single-stranded RNA virus with a 9.6 kb genome that encodes ten viral proteins. Among them, the NS3 protease and the NS5B polymerase are essential for viral replication and have been the main focus of drug discovery efforts. Aided by structure-based drug design, potent and specific inhibitors of NS3 and NS5B have been identified, some of which are in late stage clinical trials and may significantly improve current HCV treatment. Inhibitors of other viral targets such as NS5A are also being pursued. However, HCV is an RNA virus characterized by high replication and mutation rates and consequently, resistance emerges quickly in patients treated with specific antivirals as monotherapy. A complementary approach is to target host factors such as cyclophilins that are also essential for viral replication and may present a higher genetic barrier to resistance. Combinations of these inhibitors of different mechanism are likely to become the essential components of future HCV therapies in order to maximize antiviral efficacy and prevent the emergence of resistance.

Influence of White Spot Syndrome Virus Infection on Hepatopancreas Gene Expression of ‘Huanghai No.2' Shrimp(Fenneropenaeus chinensis)

To elucidate the molecular response of shrimp hepatopancreas to white spot syndrome virus(WSSV) infection, microarray was applied to investigate the differentially expressed genes in the hepatopancreas of ‘Huanghai No. 2'(Fenneropenaeus chinensis). A total of 59137 unigenes were designed onto a custom-made 60 K Agilent chip. After infection, the gene expression profiles in the hepatopancreas of the shrimp with a lower viral load at early(48–96 h), peak(168–192 h) and late(264–288 h) infection phases were analyzed. Of 18704 differentially expressed genes, 6412 were annotated. In total, 5453 differentially expressed genes(1916 annotated) expressed at all three phases, and most of the annotated were either up-or down-regulated continuously. These genes function diversely in, for example, immune response, cytoskeletal system, signal transduction, stress resistance, protein synthesis and processing, metabolism among others. Some of the immune-related genes, including antilipopolysaccharide factor, Kazal-type proteinase inhibitor, C-type lectin and serine protease encoding genes, were up-regulated after WSSV infection. These genes have been reported to be involved in the anti-WSSV responses. The expression of genes related to the cytoskeletal system, including β-actin and myosin but without tubulin genes, were down-regulated after WSSV infection. Astakine was found for the first time in the WSSV-infected F. chinensis. To further confirm the expression of differentially expressed genes, quantitative real-time PCR was performed to test the expression of eight randomly selected genes and verified the reliability and accuracy of the microarray expression analysis. The data will provide valuable information to understanding the immune mechanism of shrimp's response to WSSV.

The Role of NK Cell in T Cell Recruitment in Murine Liver Infected with Adenovirus

To study the role of natural killer (NK) cells in T cell recruitment in murine liver infected with virus, mice were intravenously injected daily with anti-NK1.1 + antibody to deplete NK cells. Lymphocytes in the liver tissue of mice infected with type 5 adenovirus depleted in the E1 and E3 regions were assessed by fluorometric activated cell sorting (FACS). Expression of chemokine IP-10 and its receptor CXCR3 mRNA in the liver, hepatic lymphocytes and spleen tissue were examined by reverse transcription polymerase chain reaction (RT-PCR). Serum alanine aminotransferase (ALT) was measured as an indicator of liver injury. It was found that infection of adenovirus and anti-Fas monoclonal antibody (mAb) into mice caused liver injury and high expression of interferon-γ inducible protein-10 (IP-10) mRNA in the liver. Anti-NK1.1 + mAb, which was intraperitoneally injected into the mice infected with adenovirus, suppresses T cell recruitment and expression of IP-10 mRNA in the liver. Slighter liver injury was also observed. After virus infection, expression of CXCR3 mRNA in spleen and liver tissue was observed at different time. The results suggested that T cell recruitment was initiated by NK cell dependent chemokine IP-10, which induced activated T cells priming in the spleen to the liver of the mouse. NK cells played a key role in T cell recruitment in the liver of mouse infected with adenovirus.

An annual topic highlight: Alcohol and liver, 2011

An annual topic highlight: Alcohol and Liver, 2011, covers the important and new aspects of pathogenesis of alcoholic liver diseases (ALD). It includes broad topics ranging from the exacerbation of ALD by infectious (viral) agents (hepatitis C virus and human immunodeficiency virus) to the influence of alcohol on liver fibrogenesis, lipid rafts, autophagy and other aspects. This issue is recommended for both basic scientists and clinicians who are involved in alcoholic liver research.

Retromer localizes to autophagosomes during HCV replication

Dear Editor,Hepatitis C virus（HCV）is a positive-strand RNA virus that belongs to the genus Hepacivirus within the Flaviviridae family.HCV causes chronic liver diseases,and185 million people are infected（Messina et al.,2015）.Currently,there is no approved vaccine to prevent hepatitis C.HCV induces autophagy through elevating reactive oxygen species（ROS）levels via the unfolded protein response （UPR） or via direct interference with the autophagic pathway.