Objective To construct the prokaryotic recombinant plasmids carring truncated HBcAg gene, whole-length HBcAg gene and HBc-HBsAg fusion gene,and to observe the expression of target proteins in E.coli and their immunogenicity in vitro. Methods Truncated HBcAg gene,whole-length HBcAg gene and HBc-HBsAg fusion gene were obtained from plasmid pUCmT-HBV containing whole-length HBV gene（subtype adr）and construct recombinant plasmids of pSK-HBs,pSK-HBc and pKS-HBV C. Truncated HBcAg gene,whole-length HBcAg gene and HBc-HBsAg fusion gene which were obtained by fusing truncated HBsAg and truncated HBcAg gene,were subcloned into a expression vector pET-30a respectively after confirmed by DNA sequencing. The gene products were expressed in E.coli BL21（DE3） and identified by SDS-PAGE and Western blot. Results The prokaryotic expression plasmids expressing truncated HBcAg gene,whole-length HBcAg gene and HBc-HBsAg fusion gene were successfully constructed. High levels of HBcAg protein and HBc-HBsAg fusion protein were observed in E.coli BL21（DE3） and their immunogenicity were confirmed by Western blot. Conclusion HBcAg protein and HBc-HBsAg fusion protein are successfully obtained by selected expressing vector in E.coli BL21（DE3） and the gene products have immunogenicity. This study has provided an experimental basis for specific immunotherapy for chronic hepatitis B.

Objective To investigate the clinical efficacy of pegylated interferon α-2a（Peg-IFNα-2a） combined with adefovir （ADV） in treatment of patients with HBeAg-positive chronic hepatitis B（CHB）. Methods Randomized controlled trials （RCT） involving peg-IFNα-2a combined with adefovir in treatment of patients with HBeAg-positive CHB,were searched from Pubmed,Wanfang Database,Chongqing VIP database,CNKI database,etc,from January,2004 to January,2014,of which the quality were underwent rigorous assessment and data were analyzed by Stata/SE software（version 12）. Results 7 allegeable RCT （a total of 529 cases） were included. There were 261 patients receiving peg-IFNα-2a combined with ADV and 268 receiving peg-IFNα-2a alone. Systematic evaluation showed that HBV DNA negative rate in combinational therapy-treated patients with CHB at the end of week 48 was significantly higher as compared to patients receiving interferon alone[OR=1.20,95% CI=（1.01,1.43）];the HBeAg seroconversion rate at 48 w in patients receiving combinational therapy was also significantly higher than that in the controls[OR=1.24,95% CI=（1.02,1.52）];a significant bias of results was found for serum ALT normalization rate [bias_p=0.012,bias_95 CI=（1.442998,6.467852）]. Conclusions Peg-IFNα-2a combined with ADV significantly improves HBV DNA negative and HBeAg seroconversion rate in patients with HBeAg-positive CHB.

Objective To explore the changes in Thl7 cell counts and serum IL-17A levels in HIV/HCV co-infected patients. Methods Thirty HIV/HCV co-infected patients,30 HIV infected patients,30 HCV infected patients and 30 healthy subjects were selected in this study. The percentage of CD4⁺T cells and Th17 cells in peripheral blood,and IL-17A in sera were detected by FCM and ELISA,respectively. Results The percentage of CD4⁺T cells in HIV and HIV/HCV co-infected patients were （310.23±114.35）/μl and（218.42±112.47）/μl,respectively,both were significantly lower than that in the healthy controls[（735.46±121.52）/μl,P<0.05],while the percentage of CD4⁺T cells in HIV/HCV co-infected group was even lower than in HIV infected patients（P<0.05）; There was no significant difference in CD4⁺T cells between HCV infected patients[（719.47±123.72）/μl] and the controls;The percentage of Th17 and serum level of IL-17A in HIV infected patients were（2.48±0.90）% and （25.18±12.63）pg/ml,significantly lower than those in the controls [（3.95±1.23）% and（39.15±16.30）pg/ml,P<0.05];The percentage of Th17 and serum level of IL-17A in HCV infected patients were（5.48±0.90）% and（45.24±15.72）pg/ml,significantly higher than those in the controls（P<0.05）;The percentage of Th17 and serum level of IL-17A in HIV/HCV co-infected patients were（1.76±0.42）% and（16.49±7.54） pg/ml,both significantly lower than those in HIV infected patients[（2.48±0.90）% and （25.18±12.63）pg/mL,P<0.05]. Conclusions Co-infection of HCV may influence the immune response in HIV infected patients,which might lead to the involvement of Th17 and IL-17A cytokine.

Objective To investigate the common agents related to drug-induced liver injury（DILI） in our hospitalized patients and to observe the clinical efficacy of ursodeoxycholic acid in treatment of patients with DILI. Methods Two hundred patients with DILI were recruited in this study and the common agents were analyzed. The patients were divided into two groups. Patients in control group received magnesium isoglycyrrhizinate, glutathione and polyene phosphatidylcholine（one,two,and all or any combination of them） for four weeks. Ademetionine was given when hyperbilirubinemia appeared. The patients in observation group received additionally ursodeoxycholic acid for four weeks at the base treatment in the control. Results The leading agents in our patients with DILI were immunosuppressive agents（34.0%）,followed by traditional Chinese medicin（28.5%）,and other common agents were antibacterial（13.0%）,antitubercular agents（7.5%）, chemotherapeutics（7.5%）,and antipyretic analgesics（4.5%）. Liver function improved in both groups after four week treatment,while the efficacy was superior in ursodeoxycholic acid-treated patients（P<0.01）. Conclusion Immunosuppressive agents are the main causes of DILI in our hospitalized patients, and the ursodeoxycholic acid could improve the efficacy of basic treatment such as glycyrrhizin in patients with DILI.

Objective To investigate the application of intima-media thickness（IMT） of carotid artery in diagnosis of phlegm-stasis syndrome in patients with nonalcoholic fatty liver diseases（NAFLD）. Methods The patients with nonalcoholic fatty liver diseases were diagnosed and the intima-media thickness of carotid artery was detected by sonography. Results Out of 80 patients with NAFLD,there were 30 with Phlegm syndrome,30 with Stasis syndrome and 20 with indistinguishable syndrome based on the syndrome determination criteria of Chinese traditional medicine. The moderate to severe degree of fatty liver accounted for 88.9% and the patients with Phlegm syndrome for 83.3% in 54 patients with increased IMT of carotid artery;the waist circumference,basic mass index（BMI）,fasting blood glucose（FBG）,HOMA-IR and cholesterol increase significantly as compare to those in healthy persons（P<0.01）;IMT in patients with NAFLD were （1.19±0.75）mm,significantly higher than in control（0.71±0.25）mm（P<0.01）; Logistic regression analysis showed that the FBG（OR：5.48,95%CI：1.39~21.61）, total cholesterol （OR：1.14,95%CI：1.14~12.82）,BMI（OR：1.20,95%CI：1.02~1.42）,age（OR：1.02,95%CI：1.01~1.03） and Phlegm syndrome（OR：1.10,95%CI：1.03~1.17） were the independent risk factors for increase IMT in patients with NAFLD. Conclusions The IMT of carotid artery is closely related to the Phlegm syndrome and fatty liver degree in patients NAFLD,so it might be used as a parameter for determination of phlegm-stasis syndrome by Chinese traditional medicine.

Objective To analyze the clinical features,outcomes,prognosis and related risk factors of cirrhotic patients complicated with diabetes. Methods Retrospective analysis was conducted in 133 cases of cirrhotic patients complicated with diabetes and 137 random cases of cirrhotic patients without diabetes from January 2011 to December 2011;the clinical characteristics,biochemical examination,complications,classification of liver function and disease outcome were recorded and compared. Results Average age of cirrhotic patients complicated with diabetes and non-diabetic cirrhotic patients were（55.9±11.0） years and（50.2 ±12.0） years respectively,average hospital stays were（15.6±11.3） days and （12.8±8.4） days,fasting blood glucose were （9.8±5.5） mmol/L and （5.1±2.1） mmol/L,aspartate aminotransferase （AST） were（83.2±98.6） u/L and（65.3±88.8） u/L,albumin （ALB） were （29.1±4.7） g/L and （30.5±5.5） g/L,triglycerides were （1.1±0.9） mmol/L and （0.7±0.4） mmol/L,and serum sodium were （134.3±5.1） mmol/L and（135.8 ± 5.7） mmol/L,respectively （P<0.05 for all）. In patients with diabetes,the numbers of alcoholic causes,Child-Pugh C grade,ascites,spontaneous bacterial peritonitis,hepatic encephalopathy and hypertension were significantly higher than those without diabetes（11.3 vs 3.7%,26.3 vs 11.3%,36.8 vs 29.9%,33.8 vs 22.6%,18.0 vs 9.0% and 21.1 vs 2.9%,respectively,P<0.05 for all）;Meanwhile,the mortality during hospitalization of patients with diabetes was significantly higher than that of patients without diabetes（18.8 vs 9.5%,P<0.05）. The Logistic analysis revealed that advanced age,duration of hospitalization,hypertension,hyperglycemia and triglyceride were the risk factor for diabetes in cirrhotic patients（odds ratio were 1.054, 1.052,16.182,1.503 and 1.503 respectively,P<0.01 for all）. Conclusion Compared with cirrhotic patients without diabetes,liver function and prognosis of cirrhotic patients with diabetes was poorer. Diabetes may contribute to the poor prognosis of patients with cirrhosis.

Objective To evaluate the accuracy of predicting esophageal variceal bleeding （EVB） in patients with liver cirrhosis with three-dimensional reconstruction by multi-slice spiral CT portography （MSCTP） and hemodynamic determination of portal vein by B ultrasonography. Methods The clinical data of 60 patients with cirrhosis who were simultaneously examined by MSCTP and B ultrasonography were collected in this study. The gastroscopy was conducted in patients with EVB. Results In 60 with liver cirrhosis, the actual number of bleeding was 28. When LGV>0.61 cm and PBF>1098.36 ml/min were determined as the cut-off value for EVB, the accurate rates by MSCTP and ultrasonography were 89.29% and 60.71%, respectively（x²=6.095,P=0.029）;26 out of 28 patients with EVB was confirmed by gastroscopy, and no esophageal varices bleeding in the other 2 cases;MSCTP and gastroscopy had no significant difference in predicting EVB （P=1.000） as determined by McNemar test,and Kappa coefficient analysis showed substantial consistency of these two methods （matching coefficient K=0.781,P=0.000）;While there was a significant difference between ultrasonography and gastroscopy in predicting EVB（P=0.012）,and Kappa coefficient analysis showed slight consistency of these two methods （matching coefficient K=0.038,P=0.747）. Conclusion MSCTP can predict EVB,and has a good consistency with gastroscopy.

Objective To observe the effectiveness and safety of combination therapy and optimized therapy of lamivudine and adefovir dipivoxil in patients with hepatitis B virus-related decompensated cirrhosis. Methods 75 patients with hepatitis B virus-related decompensated liver cirrhosis received either combination therapy or optimized therapy of lamivudine and adefovir dipivoxil from 2010 to 2013,and the clinical effectiveness and safety after 48-week treatment in two groups were compared. Results At the end of 48 week treatment,the normalization rate of serum ALT and the loss of serum HBV DNA in combination therapy group were 84.0% and 89.3%,respectively,much higher than those（46.0% and 50.7%,respectively,P<0.05） in patients receiving optimized therapy;the number of Child-Pugh class A increased by 22.7% and that of Child-Pugh class C reduced by 12% in combination therapy group,greater than 10.7% and 5.3% in optimized therapy group（P<0.05 for both）;the ascites subsided in 76.0% of patients in combination therapy group,significantly higher than 56.0% in optimized therapy group（P<0.05）;there was no difference in changes of renal function index and plasma creatinine kinase throughout the treatment between the two groups. Conclusions The combination of lamivudine and adefovir dipivoxil offers a better clinical outcome in patients with hepatitis B virus-related decompensated cirrhosis.

Objective To observe the changes of hepatic hemodynamics by ultrasonography before and after splenectomy in patients with hepatolenticular degeneration（HLD） complicated by hypersplenism. Methods 108 patients with HLD were recruited and they all received color doppler ultrasonography before and after splenectomy. The inner diameter and maximum velocity of right portal vein （Vmax）,peak systolic velocity （PSV） and resistance index（RI） of right hepatic artery were recorded and analyzed. Results The inner diameters of right portal vein before and after splenectomy were （9.09±2.16） mm and （8.48±1.88） mm,respectively（P>0.05）;Vmax of right portal vein after splenectomy declined remarkably as compared with that before splenectomy[（19.40±3.41） cm/s vs.（23.47±5.78） cm/s,P<0.01];PSV and RI of right hepatic artery after splenectomy were significantly higher than those before surgery [（60.71±11.85） cm/s vs. （40.33±8.83） cm/s and （0.66±0.05） vs. （0.63±0.05）,P<0.01 for both]. Conclusion Prominent changes in blood supply of portal venous system and hepatic artery system in patients with HLD and hypersplenism take place after splenectomy. Ultrasonography can monitor the changes of hepatic hemodynamics and provide information for the evaluation of patients' liver function.

Objective To observe the efficacy and safety of transcatheter arterial chemoembolization （TACE） combined with targeted therapy for advanced hepatocellular carcinoma. Methods From Jan. 2008,to Dec. 2012,60 cases of advanced hepatocellular carcinoma received TACE,among them,37 cases were treated with sorafenib after TACE,at a dose of 400 mg,twice a day（the dosage was adjusted according to the side effect）. Abdominal CT scans were regularly performed and treatment efficacy was evaluated according to the modified RECIST. The side effect of sorafenib was recorded,so was the liver function before and after TACE. Results In 37 patients received combined treatment（TACE plus sorafenib） and 23 patients received TACE alone,the median overall survival time were （13±0.98） months and （5±0.62） months,respectively （P=0.001）,the time to progression were （7.5±1.21） months and （5±0.62） months,respectively （P=0.001）,and at the end of followed-up,the disease control rate were 48.6% and 17.4% in the two groups,respectively（P<0.01）;The multivariate analysis showed that the risk factors for death were without combination of sorafenib and antiviral therapy,and the presence of portal vein tumor thrombus. The main side effect of sorafenib was hand-foot skin reaction. Conclusion TACE combined with sorafenib has prolonged time to disease progression and overall survival time in patients with advanced hepatocellular carcinoma with relatively good safety.

Objective To investigate the impact of pirfenidone on liver fibrosis induced by bovine serum albumin （BSA） in rats and its relevant mechanism. Methods 30 male Wistar rats were randomly divided into three groups: e.g. in control group the rats were given saline intraperitoneally,in model group were given BSA intraperitoneally,and in treatment group were given intragastric administration of pirfenidone at the dose of 200 mg.kg^-1.d^-1 after 5-week intraperitoneal injection of BSA. The animals were sacrificed after 4-week treatment and the liver specimens were collected. Hepatic pathology was performed by haematoxylin-eosin stain and Masson trichrome stain,and the expression of Smad 6 was detected by immunohistochemistry. Results As compared with in model group, the number of cases with liver fibrosis stage of S3-S4 in treatment group decreased, with significantly reduced infiltration of inflammatory cells,narrowed fibrous septum in liver tissues. Relative expression of Smad 6 in model group was significantly lower than that in treatment group [（108.2±33.6） vs. （329.4±39.2）,P<0.05]. Conclusions Pirfenidone can inhibit liver fibrosis induced by BSA in rats,which might be attributed to the upregulation of Smad 6.

Objective To observe the effects of transforming growth factor β type I receptor （TβRI） antisense RNA and tissue inhibitor of metalloproteinase 1（TIMP-1） antisense RNA on experimental liver fibrosis in rats. Methods TβRI and TIMP-1 antisense RNA eukaryotic expression plasmids were constructed and transfected into experimental rats with hepatic fibrosis. The impacts of TβRI and TIMP-1 antisense RNA on TIMP-1,TβRI and MMP-13 expression were detected by Western blot. Results The relative expressions of TIMP-1 protein in normal control group, antisense TβRI group, antisense TIMP-1 group, combinational group, plasmid group and model group were（23.6±2.8）,（57.96±3.8）,（62.3±2.8）,（34.2±2.8）,（279.2±29.8）and（287.3±23.7）,respectively;the relative expressions of TβRI in each groups were（56.2±2.7）,（70.5.±1.8）,（77.0±1.7）,（60.6±2.2）,（232.0±19.4） and（241.0±18.3）,respectively;and the relative expressions of MMP-13 were （17.84±2.3）,（36.2±3.7）,（41.3±2.4）,（28.6±2.0）,（127.3±1.2）and（118.5±2.5）,respectively. The relative expressions of TβRI,TIMP-1 and MMP-13 in antisense TβRI group, antisense TIMP-1 group and combinational group were significantly decreased compared with normal control group and plasmid group（P<0.05）,while there was no significantly differences between plasmid and model group. Conclusions Antisense TβRI RNA and antisense TIMP-1 RNA can both inhibit the expressions of TβRI,TIMP-1 and MMP-13 protein effectively,and their combination can improve the inhibiting effects.

The clinical manifestations of Wilson's disease appear to be variable and it is sometimes difficult to diagnose at early stage. Although it is a genetic disorder of copper metabolism, if discovered early,effective treatments are available and the prognosis is good. The management of this disease in pregnant women, children and patient with acute liver failure,is highly concerned in clinical practice.

Endoplasmic reticulum（ER） homeostasis is perturbed by various pathological conditions,and a given new synthesized unfolded proteins might accumulate in the ER and result in endoplasmic reticulum stress（ERS）. ERS is related to the pathogenesis of the various liver diseases,and some scholars believe that the intervention of ERS will provide new choices for prevention and treatment of liver diseases. This paper will review the unfolded protein response due to ERS,which might be the target for medical intervention for liver diseases.

Transplantation of mesenchymal stem cells（MSCs） is an promising way in the treatment of cirrhosis and liver failure. The possible mechanisms of MSCs in repair of liver injury are as follows：differentiation to hepatocyte-like cells,paracrine and immunomodulatory effects,and signal transmission mediated by microvesicles （MVs）. Results from present studies suggest that RNA carried by MVs has emerged as an important mechanism by which the stem cells may repair injured cells and MVs transplantation may become a new choice for improving decompensated liver functions. This review will discuss the possible mechanisms of intrahepatic transplanted MSCs and the potential of MVs in the treatment of liver injuries.

Hepatic fibrosis （HF） is a pathological process in a variety of chronic liver diseases, which may eventually progressed to cirrhosis or even liver cancer. Therefore,how to delay,cease or reverse HF is particularly important. HF is a complicated process that is involved by various cytokines and cell signaling pathways. Elucidation of these signaling pathways might provide some approaches to intervene the development of HF.

Hepatitis B virus （HBV）-related acute-on-chronic liver failure （ACLF）,characterized by rapid disease progression and high mortality,is a serious health problem in China. The outcomes will differ if the clinicians can diagnose the patients with ACLF in time and give them instant intervention. So some scholars in the world began to study the patients in so-called pre-liver failure stage. The scholars in the west,Hong Kong and Taiwan defined it as severe acute exacerbation of chronic hepatitis B,while the scholars in the mainland tried to set up three different diagnostic criteria for pre-ACLF. This review mainly focuses on recent advances in the definition, diagnosis and treatment of patients with HBV-related pre-ACLF.

Hepatitis C virus （HCV） is a single-stranded RNA virus that is about 9.6 kb. Its open reading frames encoding structural proteins and non-structural proteins. HCV infection is approximately 3% worldwide. It is one of the most serious infection diseases that threaten human health. Screening tests in blood donors is the most effective means to prevent the communication of HCV. The current methods for screening tests include detection of HCV antigen,anti-HCV antibody,HCV antigen-antibody complex,and HCV genes. Antigen detection is the initial screening test and it detects seroconversion after a long infectious window period. The detection of HCV core antigen might shorten the window period and has a high sensitivity,but interference by immunoglobulins in vivo influence its widespread uses. Rapid tests is simple to perform,but its sensitivity is low. Molecular testing for HCV RNA utilizing nucleic acid amplification technology （NAT） is the most sensitive assay and can shorten the window period,while the costs is very expensive. Each screening tests for HCV infection has its advantages and disadvantages. Currently,the blood centers are using at least two different reagents to detect HCV antibodies for blood screening.

Nucleos（t）ide analogs（NAs） have been successfully used for treatment of chronic hepatitis B. Hepatitis B virus（HBV） replication is now recognized as the key driverof liver injury and disease progression, so the primaryaim of treatment forchronic HBV infection is to maximize sustained suppression of HBV replication to undetectablelevels. The long-term treatment has also beenshown to achieve substantial histological improvement and regression of liver fibrosis orcirrhosis,and reduction of hepatocellular carcinoma. This paper has reviewed the necessity, clinical benefits, and the management of long-term treatment for chronic hepatitis B.