Objective To construct human KCTD9 protein in sf9 expression system and harvest the protein for further functional study. Methods To express hKCTD9 in insect cells，the full length hKCTD9 gene was obtained from pMD18-T-hKCTD9 plasmid. The gene was then inserted into pENTR/D-TOPO plasmid. After homologous recombination of the plasmid with BaculoDirectTM linear DNA，the recombinant baculovirus DNA　containing the hKCTD9 gene fragment was obtained. Sf9 cells were transfected with baculovirus DNA via Cellfectin and cultured for 3 days. After 3 rounds amplification of recombinant baculovirus，the expression of hKCTD9 in sf9 cells was detected and identified by the laser confocal microscopy and Western blot respectively. And finally the baculovirus particle were catched by transmission electron microscopy（TEM）. Results PCR confirmed the identity of recombinant plasmid pENTR/D-TOPO-hKCTD9 and the recombined baculovirus containing hKCTD9 fusion gene. The laser confocal microscopy found that the hKCTD9 fusion proteins were expressed efficiently in infected sf9 cells and localized in the nuclei of sf9 cells. The expression hKCTD9 fusion protein was 55KD as shown by Western blot，other than 43KD in gene bank. TEM photograph showed that numerous recombinant baculocirus located in nuclei and cytoplasm in sf9 cells. Conclusion Human KCTD9 protein is expressed successfully using baculovirus expression system，which provides the basic tool for its further functional study.

Objective To observe the inhibition of hepatitis B viral replication in HepG2215 cells by siRNAs targeting hepatitis B virus X gene. Methods Four siRNAs against the X region of hepatitis B virus were chemically synthesized，and then were subjected to HepG2.2.15 cells by liposome-mediated transfection. The HBsAg and HbeAg in the supernatant were assayed by ELISA and HBV DNA by RT-PCR. Results At 30nmoL/L，the four siRNA had no obvious suppression in HBeAg and HbsAg expression（P>0.05），while at the concentration of 60nmoL/L and 90nmoL/L，the inhibition of siRNA-1 and siRNA-4 were statistically significant with the inhibition rates of 41% and 43%；the HBV DNA replication was obviously reduced. Conclusion The chemically synthesized siRNAs can effectively inhibit hepatitis B viral replication in vitro.

Objective To study effects of glycine on reducing blood lipids and protecting liver in mice with hyperlipidemia. Methods 40 mice were randomly divided into control，high and low fat diet and glycine-treated groups. Total cholesterol（TC），triglyceride（TG），high density lipoprotein cholesterol（HDL-C），low density lipoprotein（LDL）levels and arteriosclerosis index（AI）were measured after 2 weeks. The activity of superoxide dismutase（SOD）and liver malondialdehyde（MDA）content in livers were also measured. Results TC，LDL，HDL-C，AI and MDA content in high fat diet group increased significantly as compared with in control（P＜0.01）；as compared with high fat diet group，the activity of SOD in liver increased（P＜0.05），MDA in liver decreased（P<0.05）and plasma TC and LDL decreased（P＜0.01） in glycine-treated groups；the hepatic steatosis improved obviously also in livers of glycine-treated mice. Conclusion Glycine can regulate lipid metabolism and enhance the antioxidation to prevent hyperlipidemia and hepatic steatosis.

Objective To investigate the polymorphisms of promoter at -238 and -308 of necrosis factor-α gene in patients with HBeAg-positive chronic hepatitis B. Methods 203 HBeAg-positive chronic hepatitis B patients were recruited and the polymorphisms in TNF-α promoter at positions -238 and -308 were determined by polymerase chain reaction-restriction fragment length polymorphism assay，and the serum levels of TNF-α were examined by ELISA. Results The frequencies of TNF-α-238G/G and G/A genotypes were 84.7% and 15.3%， respectively，and the frequencies of TNF-α-308G/G，G/A and A/A genotypes were 76.8%，22.7% and 0.5%， respectively；The levels of TNF-α in patients with TNF-α-238G/A genotype were significantly lower than in those with G/G genotype（201.2±36.3 pg/ml vs 215.7±34.7pg/ml），（x2=4.355，P=0.037），and the levels of serum TNF-α in patients with -308G/A genotype were significantly higher than in those with G/G genotype（234.6±37.5pg/ml vs 207.4±32.3 pg/ml），（x2=14.653，P＜0.001）. Conclusion Higher levels of serum TNF-α are found more often in patients with TNF-α-238G/G or -308G/A genotypes.

Objective To explore the clinical characteristics between patients with M204 and with M204/A181T mutation in HBV reverse transcriptase（rt）region. Methods Eighty-seven patients with rtM204 site mutation and forty-one patients with rtM204/rtA181T mutation were selected. The levels of HBsAg，HBV DNA and ALT were analyzed with statistic method. All the treatment history with nucleos（t）ide analogues（NA）were reviewed retrospectively. Results Compared to rtM204 site mutation, rtM204+rtA181T mutation did not lead to the decline of HBsAg（P=0.3020）and HBV DNA（P=0.5917）and higher ALT level（P=0.8001）；Moreover，the mono-therapy of lamivudine was easy to induce rtM204 site mutation（41.38%），while lamivudine swicthing to adefovir was easy to induce the rtM204+rtA181T mutation（34.15%）. Conclusion The NAs for patients with rtM204 site mutation and for patients with rtM204+rtA181T mutation were not the same，but the virological characteristics between them were not significantly different.

Objective To observe the efficacy of adefovir on patient with CHB. Method 42 CHB patients were divided into two groups based on the ratio of CD4+/CD8+. Two groups received adefovir for 12 months. Results After 12 months treatment，the high level group got a better improvement in the liver function，HBV DNA loss and HbeAg negative rate. Conclusion Pre-treatment levels of T lymphocyte subgroup in CHB will influence the efficacy of adefovir dipivoxil treatment.

Objective To investigate the relationship between estrogen receptor α gene polymorphisms and therapeutic response to interferon-alpha in patients with chronic hepatitis B. Methods PvuII polymorphisms of estrogen receptor α（ESR1）gene were analyzed in 100 naive patients with chronic hepatitis B receiving interferon-alpha treatment by polymerase chain reaction-restriction fragment length polymorphism technique. Results Among PP，Pp and pp genotype group，the early virological response were markedly higher in PP genotype groups （70.8%） than 40.0% in Pp and 37.2% in pp genotype group（P＜0.05），while neither the combined response nor HBeAg disappearance or seroconversion among PP，Pp and pp genotype groups had no significant difference（P＞0.05）. Conclusion PvuII genotype polymorphism of ESR1 is associated with early virological response to IFN-α in patients with CHB and the detection of PvuII genotype polymorphism might be helpful for predicting early response to IFN-α treatment.

Objective This study aims to observe the curative effect and safety of auto-marrow stem cells in treatment of patients with decompensated liver cirrhosis. Methods 97 cases with decompensated liver cirrhosis were randomly divided into control group（with conventional therapy）and therapy group（with treatment of auto-marrow stem cells on the base of conventional therapy）. The clinical effectiveness，blood biochemical index and adverse events after 1，2，3，4，8 weeks were observed. Results All patients in therapy group had obviously their improvement in clinical symptoms after treatment，while only 4.2% patients improved in control group；The level of albumin after 8 weeks was 33.1±8.3g/L in study group，much higher than 25.6±7.6g/L in control group（P<0.05）；The prothrombin time activity were 43.0±11.4% and 38.2±12.5% in the two groups，respectively（P<0.05）；The TBIL were 38.3±12.2 μmol/L and 40.6±13.3 μmol/L，respectively（P>0.05）；The level of AFP， leukocyte and platelet counts had no difference between the two groups. Conclusion The curative effect of auto-marrow stem cells in treatment of patients with decompensated liver cirrhosis is safe and effective.

Objective To observe the effect of interferon-alpha，lamivudine and oxymatrine treatment for hepatitis B-induced hepatic failure survivals． Methods One hundred and ten subacute or acute-on-chronic hepatic failure survivals（group A） received interferon alpha（n=39），lamivudine（n=37），or combination of lamivudine and oxymatrine（n=34）therapy for six months，and one hundred and ten chronic hepatic failure survivals（group B）received lamivudine（n=53），or combination of lamivudine and oxymatrine（n=57）therapy for six months，respectively． All patients were followed-up for eighteen months. Results At the end of follow-up，all patients in group A survived and 25.6%（10/39）of survivals with interferon alpha treatment switched to lamivudine therapy，and 21.1%（15/71）with lamivudine or lamivudine and oxymatrine to adefovir dipivoxil therapy，while in group B five patients （9.4％）receiving lamivudine，and six（10.5％）patients receiving combinational therapy died and 18.2%（18/99）of survivals switched to adefovir therapy；80% of survivals with lamivudine treatment in both groups had their serum HBV DNA loss and the HbeAg negativity and/or HBeAg/anti-Hbe seroconversion rates in patients with interferon alpha or lamivudine and oxymatrine treatment were not higher than those in survivals with lamivudine alone. Conclusion Long term administration of nucleos（t）ides analogues might block the progress of hepatic fibrosis in hepatitis B-induced liver failure survivals，however，the effects of interferon alpha or matrine treatment in this settings maybe due to relatively short-term administration needs to be further investigated.

Objective To evaluate the curative effect of plasma exchange in the treatment of severe hepatitis by the model for end-stage liver disease. Method 62 patients with severe hepatitis were treated with plasma exchange. Blood chemical data were collected pre-treatment and post-treatment. All of the patients graded with MELD and the fatality rate were observed three months later. Results The concentration of TBil，TBA，ALT in the sera were much lower than post-treatment（P＜0.05）；After plasma exchange treatment，overall death rate of 62 patients was 56.5%，much less than the prediction of mortality（76.3%，P<0.05），especially in the group of patients with MELD score less than 30. Conclusion As a hepatic support device，plasma exchange is a useful way for therapy of severe hepatitis.

Objective To explore the characteristics of renal artery blood flow parameters at all levels in patients with cirrhosis. Methods Renal artery blood flow parameters at all levels were detected by the use of color Doppler ultrasound in 46 patients with compensated，56 with decompensated liver cirrhosis and in control. The renal artery （MRA），segmental arteries（SRA），interlobar artery（IRA）were selected to detect. Results Compared with control group，Ri，PI，Vmax and Vmix in compensated cirrhosis were not significantly different（P>0.05）， but the same indexes in decompensated cirrhosis group changed with obvious difference（P<0.01）. Blood flow velocitis of renal artery in different levels of patients with decompensated cirrhosis decreased. Conclusions The detection of renal artery blood flow parameters would provide a valuable basis in understanding the kidney disease progression.

Objective To evaluate the clinical features，laboratory tests in male patients with primary biliary cirrhosis（PBC）. Methods The clinical data of 19 cases were analyzed retrospectively. Results 17 cases（89%）were middle-elderly aged with mean age of 54±17yr；7 cases（36.7%）were asymptomatic，and 9 cases（47.7%）were decompensated hepatic cirrhosis；The AMA in 13 patients（68.4%）and AMA-M2 in 14（73.7%）were positive；With ursodeoxycholic acid treatment，patients in early stage improved. Conclusion Male patients with PBC have relatively mild clinical symptoms and low autoantibodies，which make the early diagnosis even hard.

Objective To observe the therapeutic effects of Chinese herbal medicine Hanfangji compound in patients with cirrhosis caused by hepatitis B virus. Methods Fifty-eight patients（treatment group）with cirrhosis caused by hepatitis B virus were orally administered with Hanfangji compound for 3 months. 28 cases were orally administered Huganpian. Results After treatment，all of the parameters including ALT，TSB，ALB，HA and PⅢP improved at some degree in the treatment group，but not in the control；the hepatic histological changes also improved. Conclusions Hanfangji compound is very effective for treatment of patients with cirrhosis caused by hepatitis B virus.

Objective To evaluate clinical therapeutic effectiveness of stereotactic conformal radiotherapy （body gamma knife）combined with transcatheter arterial chemoembolization（TACE）for primary hepatic carcinoma（PHC）. Methods A total of 120 patients with unresectable PHC were randomly divided into two groups. 58 patients were treated with TACE，while 62 patients with TACE combined with stereotactic conformal radiotherapy. Results There were 7CR，28PR，16NC and 7PD in the group treated with TACE and 10CR，39PR，11NC，2PD in the group treated with TACE combined with stereotactic conformal radiotherapy；The group treated with TACE had a median survival duration of 9.3 months and 12.5 months in the group treated with TACE compared with stereotactic conformal radiotherapy. Conclusion The stereotactic conformal radiotherapy technique combined with transcatheter arterial chemoembolization is a safe and effective treatment for PHC and improve the survival of PHC patients．

Objective To investigate the differential expression of somatostatin receptor subtypes（SSTR）and the major subtypes in hepatocellular carcinoma（HCC）. Methods Immunohistochemistry were used to detect protein expressions of SSTR2，3，5 in 50 HCC and 24 normal liver tissue，and SSTR mRNA were detected in 10 HCC tissues by RT-PCR. Results The positive rate of SSTR2，3，5 protein in HCC tissues were 72%，34%，14%，respectively；The expression of SSTR2 in HCC were markedly higher than in non-HCC liver tissues；The positive rate of SSTR2，3，5 mRNA were 100%，80% and 30%，higher than in non-HCC liver tissues. Conclusion The expression of SSTR subtype is higher in HCC than in non-HCC liver tissues. SSTR2 is the main subtypes expressed in HCC tissues.