Objective To determine whether HBx gene can directly induce hepatocellular carcinoma in vivo, and to explore the mechanism of transplantation tumor in nude mice.Methods pCMVX/QSG7701 cell lines were vaccinated into subcutaneous tissue of nude mice. pRcCMV2/QSG7701 and QSG7701 cell lines were used as controls. The sections of transplantation tumor were observed microscopically by HE staining. RT-PCR was used to detect the expression of mutant p53 and c-Myc mRNA in transplant tumor and an other 3 cell lines. Results The transplant tumor occurred within the subcutaneous tissue of the nude mice inoculated with pCMVX/QSG7701 cell lines at 2nd week after the vaccination. No metastatic tumor was found in other organs. Transplant tumor was not formed in all the controls. HE staining confirmed that the transplant tumor was hepatocellular carcinoma. The mutant p53 mRNA and c-Myc mRNA expression level of transplant tumor and pCMVX/QSG7701 cells was significantly higher than that of pRcCMV2/QSG7701 and QSG7701 cells, respectively (P＜0.01).Conclusion HBx gene can up-regulate the expression of mutant p53 and c-Myc genes, and directly induce hepatocellular carcinoma in vivo.

OBJECTIVE:To investigate the anti-HBV activities of deoxynojirimycin(DNJ)derivatives N—benzyl—1—DNJ(P-DNJ)and N—nonyl—1—DNJ(NN-DNJ)in vitro.METHODS:Human hepatoma carcinoma cell lines HepG2 2,2,15,which were transfected from HBV DNA,were taken as target cells,cells were cultured with different concentrations of test drugs, with HBsAg,HBeAg and HBV DNA in the cultured supernatant determined by time-resolved immunofluorometric assay and fluorescent quantitation PCR assay on day 6th and 10th day,meanwhile,with the cytotoxicity of DNJ derivatives determined by MTT assay.RESULTS:No cytotoxic effects was noted when the test concentration of P-DNJ and NN-DNJ was within 5～125?g?mL-1,HBsAg and HBeAg levels and HBV DNA secretion decreased significantly at a concentration of 125?g?mL-1.CONCLUSION:P-DNJ and NN-DNJ showed anti-HBV activity in the in vitro cell culture experiment.

Objective To investigate gene variation and the relationship between gene variation and pathogenicity of transfusion transmitted virus(TTV).Methods The TTV DNA in the serum sample from a blood donor(BD) and a chronic non-A-G severe hepatitis(CSH) patient with TTV infection was amplified by using PCR.The purified PCR product was cloned and 10 clones from each case were sequenced.The sequences were compared among different clones and analyzed by Phylogentic tree.Results There were two different TTV strains in the BD and seven different TTV strains in the chronic non-A-G severe hepatitis patient.The TTV clones in the BD were of G1a subtype and those of the CSH were of G1a and G1b subtype.Conclusion Gene variant of TTV was much more complicated in the CSH patients than that in the BD ones.

Objective:To observe the sensitivity of transcription mediated amplification (TMA),and to compare its performance with real-time reverse transcription polymerase chain reaction (real-time RT-PCR) in detecting human immunodeficiency virus RNA (HIV RNA).Methods:TMA system was established with TaqMan probes,specific primers,moloney murine leukemia virus (MMLV) reverse transcriptase,T7 RNA polymerase,and reaction substrates.The sensitivity of TMA was evaluated by amplifying a group of 10-fold diluted HIV RNA standards which were transcribed in vitro.A total of 60 plasma of HIV infected patients were measured by TMA and Cobas Amplicor HIV-1 Monitor test to observe the positive rate.The correlation and concordance of the above two technologies were investigated by linear regression and BlandAltman analysis.Results:TMA system was established successfully and HIV RNA transcribed standards at concentration of equal or more than 10 copies/mL could be detected by TMA technology.Among 60 samples of plasma from HIV infected patients,46 were positively detected and 12 were negatively amplified by both TMA and Cobas reagents;2 samples were positively tested by Cobas reagent but negatively tested by TMA system.The concordance rate of the two methods was 97.1％ and the difference of positive detection rate between the two methods was not statistically significant (P＞0.05).Linear regression was used for 46 samples which were positively detected by both TMA and Cobas reagents and showed an excellent correlation between the two reagents (r=0.997,P＜0.001).Bland-Altma analysis revealed that the mean different value ofHIV RNA levels for denary logarithm was 0.02.Forty-four samples were included in 95％ of credibility interval of concordance.Conclusion:TMA system has the potential of high sensitivity.TMA and real-time RT-PCR keep an excellent correlation and consistency in detecting HIV RNA.

Objective To investigate clinical features and antifungal therapeutic effect of chronic severe hepatitis (CSH) patients with invasive fungal infection (IFI), and to improve the diagnosis and treatment.Methods Clinical manifestation, blood routine, imageology and mycetology characteristic, antifungal treatment perscription and therapeutic effect of 79 CSH patients with IFI were retrospectively analyzed. Antifungal therapeutic effect was compared between fluconazole and voriconazole. Results Thirteen (16.5%) patients received glucocorticoid or other immunodepressants for a relatively long time, 40 (50.6%) patients had invasive operation, and 61 (77.2 %) patients were administered 1-6 kinds of broad-spectrum antibiotics. Seventy-three patients had fever. Leucocytes and neutrophilic granulocyte increased in 96.2% of the patients. Lung (31.6%), intestinal tract (26.2%) and oral cavity (14%) infections were common. Fungus was found in 70.9% of the patients. Candida albicans (40.9%) and aspergillus (21.1%) were often seen. Halo signs and crescent signs on lung CT were relatively specific in 40% of the patients with fungal pneumonia. Voriconazole was more effective than fluconazole(71.4% vs. 39.0%, P<0.05). Twelve patients with lung aspergillus infection were administered voriconazole, 8 (66.7%) patients of whom was effective, and the other 4 (33.3%) patients died. Conclusion There are high risk factors in major CSH patients with IFI. The most common clinical manifestations of CSH patients with IFI are fever, leukocytosis, lung and intestinal tract infection. Candida albicans and aspergillus infection are common. Voriconazole is more effective than fluconazole, and can increase the survival rate of CSH patients with IFI.

Background/Aims: Acute-on-chronic liver failure (ACLF) is a catastrophic illness. Few studies investigated the prognostic value of vitamin D-binding protein (VDBP) for hepatitis B virus (HBV)-related ACLF (HBV-ACLF) resulted in conflicting results. Methods: Two prospective HBV-ACLF cohorts (n=287 and n=119) were enrolled to assess and validate the prognostic performance of VDBP. Results: VDBP levels in the non-survivors were significantly lower than in the survivors (P<0.001). Multivariate Cox regression demonstrated that VDBP was an independent prognostic factor for HBV-ACLF. The VDBP level at admission gradually decreased as the number of failed organs increased (P<0.001), and it was closely related to coagulation failure. The areas under the receiver operating characteristic curve (AUCs) of the Child-Pugh-VDBP and chronic liver failuresequential organ failure assessment (CLIF–SOFA)-VDBP scores were significantly higher than those of Child-Pugh (P<0.001) and CLIF-SOFA (P=0.0013). The AUCs of model for end-stage liver disease (MELD)-VDBP were significantly higher than those of MELD (P= 0.0384) only in the case of cirrhotic HBV-ACLF patients. Similar results were validated using an external multicenter HBV-ACLF cohort. By longitudinal observation, the VDBP levels gradually increased in survivors (P=0.026) and gradually decreased in non-survivors (P<0.001). Additionally, the VDBP levels were found to be significantly decreased in the deterioration group (P=0.012) and tended to be decreased in the fluctuation group (P=0.055). In contrast, they showed a significant increase in the improvement group (P=0.036). Conclusions The VDBP was a promising prognostic biomarker for HBV-ACLF. Sequential measurement of circulating VDBP shows value for the monitoring of ACLF progression.

To explore the prognostic factors for patients with drug-induced liver failure (DILF) and construct a logistic regression model (LRM).
 Methods: A retrospective analysis of clinical data was performed in 183 hospitalized patients, who were diagnosed with DILF in Xiangya Hospital, the Second Xiangya Hospital and the Third Xiangya Hospital, Central South University from January 2009 to January 2018. The patients were divided into an improved group (n=67) and an ineffective group (n=116) according to their prognosis. Univariate analysis was performed to screen for possible prognostic factors such as age, Tbil, SCr, PT and complications. According to the results of univariate analysis, the multivariate analysis was performed to determine the independent prognostic factors and construct a LRM. The LRM was compared with the model for end-stage liver disease (MELD), the predictive value of LRM and MELD was evaluated by receiver operating characteristic curve (ROC), the parameters such as area under the ROC (AUC) and total accuracy were compared between the 2 models and verified by another independent sample.
 Results: According to univariate analysis, there was significant differences in age, clinical type, hepatic encephalopathy, hepatorenal syndrome, WBC count, the ratio of aspartic acid transaminase (AST) to glutamine transaminase (ALT) (AST/ALT), Tbil, SCr, PT and alpha-fetoprotein (AFP) between the 2 groups (all P<0.05). Multivariate analysis revealed that: AFP, PT, AST/ALT, hepatic encephalopathy and hepatorenal syndrome were independent prognostic factors for DILF, which could be applied to constructing a LRM. The AUC of LRM and MELD was 0.917 (95% CI 0.876 to 0.959) and 0.709 (95% CI 0.633 to 0.786) respectively, the total accuracy rate of prediction for the LRM and the MELD was 86.7% and 68.3% respectively, there was significant difference in AUC and total accuracy rate between the LRM and the MELD (P<0.05). LRM was superior to MELD.
 Conclusion: AFP, PT, AST/ALT, hepatic encephalopathy and hepatorenal syndrome were independent prognostic factors for DILF; the LRM can well predict the prognosis in the DILF patients, which is superior to the MELD.
China ; Humans ; Liver Failure ; chemically induced ; diagnosis ; Logistic Models ; Predictive Value of Tests ; Prognosis ; ROC Curve ; Retrospective Studies ; Severity of Illness Index