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Current therapy with nucleoside/nucleotide analogs for patients with chronic hepatitis B

Hepatobiliary & Pancreatic diseases International 2006年第3期5卷 350-359页

作者： Xiao-Wei Xu and Ya-Gang Chen department of infectious diseases, first affiliated hospital, zhejiang university school of medicine, hangzhou 310003, china department of infectious diseases First Affiliated Hospital Zhejiang University School of Medicine Hangzhou 310006 China.

BACKGROUND: Currently, more and more nucleos(t)ide analogs are appearing as therapeutic options in the treatment of chronic hepatitis B (CHB). Their efficacy and safety profile in hepatitis B virus (HBV) infection have already been studied in detail worldwide. This review summarizes the efficacy of lamivudine, adefovir, entecavir and newer antiviral agents such as emtricitabine, telbivudine and clevudine in the treatment of hepatitis B in different clinical situations. DATA SOURCES: An English-language literature search using OVID and MEDLINE was performed and a total of 40 articles on the treatment of chronic hepatitis with nucleos(t)ide analogues were selected. RESULTS: Nucleos(t)ide analogs such as lamivudine, adefovir and entecavir are well tolerated and induce a decrease in serum HBV-DNA levels associated with normalization of serum alanine aminotransferase (ALT) levels. But their sustained response with HBeAg to anti-HBe seroconversion is rarely obtained and HBsAg loss is exceptional. The response is maintained during therapy which needs to be continued indefinitely in the majority of patients since withdrawal of treatment is generally followed by a rapid reactivation of hepatitis B. However, drug resistant mutations can be induced in long-term treatment. Other newer antiviral agents such as emtricitabine, telbivudine and clevudine in the treatment of hepatitis B are still under phaseⅡorⅢclinical trials. CONCLUSIONS: Nucleos(t)ide analogs play an important role in the therapy of hepatitis B now and in the future. Lamivudine is limited by the frequent emergence of drugresistant (HBV) mutants (YMDD). Adefovir and entecavir appear to be effective against both YMDD mutation and wild type. Therapeutic options against hepatitis B virus remain a major clinical challenge.

关键词： hepatitis B antivirus lamivudine adefovir entecavir emtricitabine telbivudine clevudine

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Hepatitis C virus infection and biological falsepositive syphilis test:a single-center experience

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Hepatobiliary & Pancreatic diseases International 2011年第4期10卷 399-402页

作者： Wei-Fang Zhu,Shui-Ying Lei and Lan-Juan Li State Key Laboratory for Diagnosis and Treatment of infectious diseases Key Laboratory of infectious diseases,zhejiang Province department of infectious diseases,first affiliated hospital,zhejiang university school of medicine,hangzhou 310003,china State Key Laboratory for Diagnosis and Treatment of infectious diseases Key Laboratory of infectious diseases Zhejiang Province department of infectious diseases First Affiliated Hospital Zhejiang University School of Medicine Hangzhou 310003 China

BACKGROUND:Rapid plasma regain positive and/or treponema pallidum hemagglutination negative [RPR(+)/TPHA(-)] results were designated as biologic false-positive (BFP).There are limited data about BFP reactions against syphilis in patients with hepatitis C virus (HCV) *** study aimed to determine the prevalence of BFP reactions for syphilis in patients with HCV infection in a large sample and assess the relationship between BFP reactions and HCV ***:A total of 2656 patients with positive anti-HCV and 5600 healthy control subjects were enrolled in this *** C serology was determined by a second generation ELISA test for HCV *** serology was determined by the RPR *** subjects with reactive RPR positive underwent the TPHA *** and laboratory data were collected by trained ***:Among 2656 patients,111 (4.2%) had a reactive RPR *** the 111 patients who were subjected to reactive RPR test,30 (27.0%) showed HCV(+)/RPR(+).Of 5600 healthy controls,80 (1.4%) had a reactive RPR *** (17.5%) controls with HCV(-)/RPR(+) had a non-reactive TPHA *** represented 1.1% of all HCV-positive and 0.3% of all HCV-negative subjects (P0.05).CONCLUSIONS:The data of this study demonstrate that HCV infection is associated with a false-positive RPR *** this study BFPs were significantly more common in HCV positive patients compared to HCV-negative *** abnormality can be considered as a predictor for *** BFPs must be considered in assessing the frequency of syphilis in a

关键词： hepatitis C virus biological false-positive syphilis

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Isolation and primary culture of rat hepatocytes

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Hepatobiliary & Pancreatic diseases International 2002年第1期1卷 77-79页

作者： Xiao-Li Liu Lan-Juan Li Zhi Chen From the department of infectious diseases, first affiliated hospital, zhejiang university school of medicine, hangzhou 310003, china department of infectious diseases First Affiliated Hospital Zhejiang University of Medicine Hangzhou 310003 China

Objective: To find out an optimal condition for isola-tion and primary culture of ***: Rat hepatocytes were isolated by a two-stepcollagenase perfusion, and cultured in hepatozyme-SFM. The reduction of MTT to formazan salt was ex-amined. Supernatant medium was collected for analy-sis of alanine aminotransferase（ALT） and ***: The two-step collagenase perfusion yielded39±12×10;cells/g liver tissue with a viability of88%±2%. Fine morphology and stable urea synthesisfor one week could be achieved when hepatocytes werecultured in ***: High yield of hepatocytes can be isolatedwith two-step collagenese perfusion. Hepatozyme-SFM is suitable for sustained growth of hepatocytes.

关键词： hepatocytes isolation primary culture artificial liver

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Isolation and short term cultivation of swine hepatocytes for bioartificial liver support system

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Hepatobiliary & Pancreatic diseases International 2005年第2期4卷 249-253页

作者： Jun Li, Lan-Juan Li, Hong-Cui Chao, Qian Yang, Xiao-Li Liu, Ji-Fang Sheng, Hai-Ying Yu and Jian-Rong Huang hangzhou, china Key Laboratory of infectious Disease, Ministry of Pub- lic Health of china Institute of infectious Disease, department of infectious diseases, first affiliated hospital, zhejiang university school of medicine, hangzhou 310003, china department of infectious diseases The First Affiliated Hospital Zhejiang University School of Medicine Hangzhou 310003 China. lilj@***

BACKGROUND: The demand for the clinical use of hepa- tocytes is increasing. The aim of this study was to develop a method for procurement of high qualitative pig hepatocytes and to evaluate the state of freshly isolated and cultured hepatocytes. METHODS: The domestic extracorporeal circulating perfu- sion apparatus was used to isolate and harvest swine hepato- cytes by the two-step perfusion method with EDTA and collagenase. The viability, function and morphology of the freshly isolated and cultured cells were evaluated and ob- served by the trypan blue exclusion test, biochemical mea- surements, phase contrast microscopy and transmission electron micrography (TEM). RESULTS: The total yield of isolated hepatocytes reached to 1.5(±0.4)×l010 per liver with a viability of 92(±5)%, and the purity of hepatocytes reached to 98% Immediately after isolation, phase-contrast microscope and TEM showed that undamaged hepatocytes appeared bright, translucent and spherical in shape, with a characteristic well-contrasted border. After 24 hours, the concentrations of alanine aminotransferase (ALT), aspartate aminotrans- ferase ( AST ), lactate dehydrogenase ( LDH ), albumin (ALB), creatinine (Cr) and blood urea nitrogen (BUN) in the fluid of culture were declined significantly. CONCLUSION: This method of procuring swine hepato- cytes could get high quality cells with active metabolic function.

关键词： bioartificial liver support system porcine hepatocytes cell isolation cell transplantation

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Effects of Salvia miltiorrhiza on intestinal microflora in rats with ischemia/reperfusion liver injury

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Hepatobiliary & Pancreatic diseases International 2005年第2期4卷 274-280页

作者： Hui-Chun Xing, Lan-Juan Li, Kai-Jin Xu, Tian Shen, Yun-Bo Chen, Yu Chen, Su-Zhen Fu, Ji-Fang Sheng, Chun-Lei Chen, Jian-Guo Wang, Dong Yan, Fang-Wei Dai and Xiao-Ying Sha hangzhou, china Key Laboratory of infectious diseases, Ministry of Public Health of china, and Institute of infectious diseases, department of infectious diseases, first affiliated hospital, zhejiang university school of medicine, hangzhou 310003, china zhejiang Academy of Medical Sciences, hangzhou 310012, china department of infectious diseases First Affiliated Hospital Zhejiang University School of Medicine Hangzhou 310003 China.

BACKGROUND: Hepatic ischemia/reperfusion injury may induce intestinal microflora imbalance. Salvia miltiorrhiza is effective in promoting blood circulation and counteracting peroxidation in tissues. The aim of the present study was to determine the effects of Salvia miltiorrhiza on intestinal mi- croflora, endotoxemia, and bacterial translocation in rats with hepatic I/R injury. METHODS: Sprague-Dawley rats in specific pathogen free grade were divided into 3 groups: group I(n =6) for sham operation: groups ( n = 7) for liver ische- mia for 20 minutes and reperfusion for 22 hours. Group was also pretreated with 4 ml/day of Salvia miltiorrhiza solu- tion (250 mg/kg) by daily gavage for 7 days. The levels of serum alanine aminotransferase (ALT), aspartate amino- transferase (AST), malondialdehyde ( MDA) and supero- xide dismutase ( SOD ) in liver tissues, serum endotoxin, intestinal bacterial counts, intestinal mucosal histology and bacterial translocation were studied. RESULTS: The levels of ALT, AST, plasma endotoxin and MDA in liver tissues were decreased more markedly in group (57.57 ± 18.08 U/L, 147.57 ±40.84 U/L, 0.42 ± 0.144 EU/ml and 0. 52 ±0.19 nmol/mg-prot respectively) in group 295.9±216.92 U/L, 0.80± 0.262 EU/ml and 0.72±0.12 nmol/mg-prot; P <0.05-0.01 respectively). Liver SOD activity was increased more sig- nificantly in group (318.47±64.62 U/mg-prot) than in group U/mg-prot, P<0.05). The counts of Bifidobacteria and Bacteroides increased more significantly in group than in group but were similar to those in group I. Bacterial translocation to the kidney in group was 50% (5/10), whereas no bacterial translocation to the kidney occurred in the other two groups (P <0. 01). Ileal mucosal structure was markedly ameliorated in group as compared with group CONCLUSIONS: Salviae miltiorrhiza could partially restore intestinal microflora balance, improve intestinal mucosal integrity, and reduce bacterial translocation and plasma en- dotoxin in rats with hepatic is

关键词： Salviae miltiorrhiza hepatic ischemia/reperfusion microflora endotoxin bacterial translocation

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A proteomic analysis of allograft rejection in rats after liver transplantation

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Science china(Life Sciences) 2007年第3期50卷 312-319页

作者： ZHANG ChunChao1,2, ZHU Feng2,3, WEI JianFeng2, ZHENG ShuSen2 & LI LanJuan1 1 department of infectious diseases, The first affiliated hospital, zhejiang university school of medicine, hangzhou 310003, china 2 Institute of Surgery, Key Laboratory of Combined Multi-organ Transplantation, Ministry of Public Health, The first affiliated hospital, zhejiang university school of medicine, hangzhou 310003, china 3 department of Hepatobiliary Surgery, The Third affiliated hospital, Suzhou university, Changzhou 213003, china department of infectious diseases The First Affiliated Hospital Zhejiang University School of Medicine Hangzhou 310003 China Institute of Surgery Key Laboratory of Combined Multi-organ Transplantation Ministry of Public Health The First Affiliated HospitalZhejiang University School of Medicine Hangzhou 310003China Institute of Surgery Key Laboratory of Combined Multi-organ Transplantation Ministry of Public Health The First Affiliated Hospital Zhejiang University School of Medicine Hangzhou 310003 China department of Hepatobiliary Surgery The Third Affiliated Hospital Suzhou University Changzhou 213003 China Institute of Surgery Key Laboratory of Combined Multi-organ Transplantation Ministry of Public Health The First Affiliated Hospital Zhejiang University School of Medicine Hangzhou 310003 China department of infectious diseases The First Affiliated Hospital Zhejiang University School of Medicine Hangzhou 310003 China

In order to understand the allograft rejection in orthotopic liver transplantation (OLT), an allograft re- jection rat model was established and studied by proteomic approach. The protein expression profiles of liver tissues were acquired by fluorescence two-dimensional difference gel electrophoresis (2D DIGE) that incorporated a pooled internal standard and reverse fluorescent labeling method. The expression levels of 27 protein spots showed significant changes in acute rejection rats. Among these spots, 19 were identified with peptide mass fingerprinting using matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS) after tryptic in-gel digestion. The results of the present paper could be helpful for our better understanding of allograft rejection in organ transplantation.

关键词： A proteomic analysis of allograft rejection in rats after liver transplantation DIGE

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Parthenolide protects human lens epithelial cells from oxidative stress-induced apoptosis via inhibition of activation of caspase-3 and caspase-9

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Cell Research 2007年第6期17卷 565-571页

作者： Hangping YaoXiajing TangXueting ShaoLei FengNanping WuKe YaoNational Key Laboratory of infectious diseases, Institute of infectious diseases, first affiliated hospital, school of medicine, zhejiang university, hangzhou, chinaEye Center, Second affiliated hospital school of medicine, zhejiang university, 88 Jiefang Road, hangzhou 310009, china National Key Laboratory of infectious diseases Institute of Infectious Diseases First Affiliated Hospital School of Medicine Zhejiang University Hangzhou China Eye Center Second Affiliated Hospital School of Medicine Zhejiang University Hangzhou China

The apoptosis of lens epithelial cells has been proposed as the common basis of cataract formation, with oxidative stressas the major cause. This study was performed to investigate the protective effect of the herbal constituent parthenolideagainst oxidative stress-induced apoptosis of human lens epithelial （HLE） cells and the possible molecular mechanismsinvolved. HLE cells （SRA01-04） were incubated with 50μM H2O2 in the absence or presence of different doses ofparthenolide （10, 20 and 50μM）. To study apoptosis, the cells were assessed by morphologic examination and AnnexinV-propidium iodide double staining flow cytometry; to investigate the underlying molecular mechanisms, the expressionof caspase-3 and caspase-9 were assayed by Western blot and quantitative RT-PCR, and the activities of caspase-3 andcaspase-9 were measured by a Chemicon caspase colorimetric activity assay kit. Stimulated with H2O2 for 18 h, a highfraction of HLE cells underwent apoptosis, while in the presence of parthenolide of different concentrations, dose-depen-dent blocking of HLE cell apoptosis was observed. The expression of caspase-3 and caspase-9 induced by H2O2 in HLEcells was significantly reduced by parthenolide both at the protein and mRNA levels, and the activation of caspase-3 andcaspase-9 was also suppressed by parthenolide in a dose-dependent manner. In conclusion, parthenolide prevents HLEcells from oxidative stress-induced apoptosis through inhibition of the activation of caspase-3 and caspase-9, suggestinga potential protective effect against cataract formation.

关键词： parthenolide human lens epithelial cells apoptosis caspase-3 and caspase-9

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Advances in cell sources of hepatocytes for bioartificial liver

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Hepatobiliary & Pancreatic diseases International 2012年第6期11卷 594-605页

作者： Xiao-Ping Pan , Lan-Juan Li State Key Laboratory for Diagnosis and Treatment of infectious diseases, first affiliated hospital, zhejiang university school of medicine, hangzhou 310003, china State Key Laboratory for Diagnosis and Treatment of infectious diseases First Affiliated Hospital Zhejiang University School of Medicine Hangzhou 310003 China

BACKGROUND: Orthotopic liver transplantation (OLT) is the most effective therapy for liver failure. However, OLT is severely limited by the shortage of liver donors. Bioartificial liver (BAL) shows great potential as an alternative therapy for liver failure In recent years, progress has been made in BAL regarding genetically engineered cell lines, immortalized human hepatocytes, methods for preserving the phenotype of primary human hepatocytes, and other functional hepatocytes derived from stem cells. DATA SOURCES: A systematic search of PubMed and ISI Web of Science was performed to identify relevant studies in English language literature using the Key words such as liver failure bioartificial liver, hepatocyte, stem cells, differentiation, and immortalization. More than 200 articles related to the cell sources of hepatocyte in BAL were systematically reviewed. RESULTS: Methods for preserving the phenotype of primary human hepatocytes have been successfully developed. Many genetically engineered cell lines and immortalized human hepatocytes have also been established. Among these cell lines the incorporation of BAL with GS-HepG2 cells or alginate encapsulated HepG2 cells could prolong the survival time and improve pathophysiological parameters in an animal model of liver failure. The cBAL111 cells were evaluated using the AMC-BAL bioreactor, which could eliminate ammonia and lidocaine, and produce albumin. Importantly, BAL loading with HepLi-4 cells could significantly improve the blood biochemical parameters, and prolong the survival time in pigs with liver failure. Other functional hepatocytes differentiated from stem cells, such as human liver progenitor cells, have been successfully achieved. CONCLUSIONS: Aside from genetically modified liver cell lines and immortalized human hepatocytes, other functionalhepatocytes derived from stem cells show great potential as cell sources for BAL. BAL with safe and effective liver cells may be achieved for clinical liver fa

关键词： bioartificial liver liver failure hepatocyte liver cell source stem cell differentiation immortalization

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Determination of platelet-activating factor by reverse phase high-performance liquid chromatography and its application in viral hepatitis

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World Journal of Gastroenterology 2005年第46期11卷 7364-7367页

作者： Hong-Cui Cao, Xiao-Ming Chen, Wei Xu, department of infectious diseases, first affiliated hospital, Medical school of zhejiang university Key Laboratory of infectious diseases of Ministry of Public Health, hangzhou 310003, zhejiang Province, china department of infectious Disease First Affiliated Hospital Medical School of Zhejiang University Zhejiang Province China.

AIM： To detect the plateletoactivating factor （PAF） and the plasma or serum levels of tumor necrosis factoroa （TNF-α） malondialdehyde （MDA）, endotoxin （ET） and to discuss their significance in various types of viral hepatitis. METHODS： PAF, TNF-α, MDA, and ET levels in 60 controls, 16 cases of acute viral hepatitis, 71 cases of chronic viral hepatitis, 19 cases of severe viral hepatitis were detected by reverse phase high-performance liquid chromatography （rHPLC）, bio-assay, ELISA, thiobarbituric acid （TBA）, and limulus lysate test （LLT）, respectively. RESULTS： The rHPLC was more sensitive and specific than bio-assay （r = 0.912, P〈0.01）. The plasma levels of PAF, TNF-α, MDA, and ET in patients with viral hepatitis were higher than those in controls （P〈0.01）. CONCLUSION： rHPLC is more reliable and accurate for the detection of PAF.

关键词： Platelet-activating factor Malondialdehyde Endotoxin rHPLC Viral hepatitis

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Animal experiment and clinical study of effect of gamma-interferon on hepatic fibrosis

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World Journal of Gastroenterology 2001年第1期7卷 42-48页

作者： Hong Lei Weng Wei Min Cai Rong Hua Liu Institute of infectious diseases, first affiliated hospital. Medical school. zhejiang university, hangzhou 310003, zhejiang Province. china Institute of infectious diseases First Affiliated Hospital Medical School Zhejiang University 261 Qingchun Lu Hangzhou 310003 Zhejiang Province China. syliao@***

AIM: To evaluate the antifibrotic effect of different doses of recombinant human Gamma-Interferon (IFN-gamma) in two rat models of hepatic fibrosis, and to observe its effect on moderate chronic hepatitis B virus fibrosis. METHODS: Hepatic fibrosis was successfully induced in 150 and 196 rats by subcutaneous injection of carbon tetrachloride (CCl4) and intraperitoneal injection of dimethylnitrosamine (DMN), respectively. Each of the two model groups was divided into: (1) fibrotic model group; (2) colchicine treatment group (0.1 mg/kg/day, gastrogavage for 8 weeks); (3) high-dose IFN-gamma group (15 MU/kg per day, i.m. for 8 weeks); (4) medium-dose IFN-gamma group (5 MU/kg daily, i.m. for 8 weeks); and (5) Y low-dose IFN-gamma group (1.67 MU/kg daily, i.m. for 8 weeks). Another group of 10 rats without any treatment was used as normal controls. At the end of the experiment, semi-quantitative histopathological scores of inflammation and fibrosis, liver alpha smooth muscle actin (alpha-SMA) expression level, liver hydroxyl proline content and serum hyaluronic acid levels were compared. And 47 medium chronic hepatitis B viral fibrosis patients were studied. They were given IFN-gamma treatment, 100 MU/day i.m. for the first three months and 100 MU qod i.m. for the next six months. Semi-quantitative pathological scores of inflammation and fibrosis and serum hepatic fibrosis indices were compared within the 9 months. RESULTS: In animal experiment, the pathological fibrosis scores and liver hydroxyl proline content were found to be significantly lower in rats treated with different doses of IFN-gamma as compared with rats in fibrotic model group induced by either CCl4 or DMN, in a dose-dependent manner. For CCl4-induced model, pathological fibrosis scores in high, medium and low doses IFN-gamma groups were 5.10 +/- 2.88, 7.70 +/- 3.53 and 8.00 +/- 3.30, respectively, but the score was 14.60 +/- 7.82 in fibrotic model group. Hydroxyl proline contents were 2.83 +/- 1.18, 3.59

关键词： Animals Antineoplastic Agents dosage Biopsy Carbon Tetrachloride Dimethylnitrosamine Disease Models, Animal Female Hepatitis B, Chronic Humans Hyaluronic Acid Hydroxyproline Interferon-gamma, Recombinant Liver Liver Cirrhosis Liver Function Tests Male Rats Rats, Sprague-Dawley

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