Objective To study the expression of Notch signaling pathway influenced by ethanol in C3A cells in vitro. Methods C3A cells were served as the control group,and the CYP2E1 cells were as experiment group. Equal concentration of ethanol and Notch signaling pathway inhibitor were added to the two groups of cells. RT-PCR and Western blotting were used to testify the expression of Notch1,NICD1 and HES1,respectively. MTT assay was used to evaluate the viability of cells. Results The cell viability in the two groups was decreased by ethanol,while the Notch signaling pathway inhibitor improved the viability of cells; The expression of NICD1 and HES1 was up-regulated in the experimental group,while when Notch signaling inhibitor was added into the medium,the expression of NICD1 and HES1 was significantly down-regulated; The Notch1 mRNA level was also up-regulated in the experimental group. Conclusion Ethanol can activate the expression of Notch signaling pathway in C3A models.

Objective To study the effect of endotoxin on intestinal mucous injury in rats with alcohol-induced liver disease. Methods 15 patients with alcoholic liver disease and 15 healthy persons were included in this study.The serum endotoxin was detected by ELISA，and serum MDA was detected by spectrophotometric method. Twenty c57/BL mice were randomly divided into model and control group（n=10 each）. The control group was fed with Lieber-Decarli liquid without ethanol，and mice in the model group were fed with 4% ethanol of Lieber-Decarli liquid. The serum endotoxin was examined after 10 weeks，and the intestinal and liver tissues were assessed with HE staining. Results The serum endotoxin in alcoholic liver disease group and control group were 0.52±0.54 Eu/L and 0.47±0.34Eu/（P<0.05），respectively;The serum MDA in alcoholic liver disease group and control group were 5.6±5.0nmol/ml and 3.7±3.4nmol/ml（P>0.05），respectively;The serum endotoxin in the model rats and control rats were 0.38±0.05 Eu/L and 0.13±0.02 Eu/L（P<0.01），respectively;The histopathological scores of colon injuries in the model group was much higher than that in control group（10.3±1.3vs 4.8±1.2，P=0.01）. Conclusion Alcoholic liver disease is always complicated by intestinal mucous injury. Endotoxemia and intestinal mucosal injury has certain correlation in alcoholic liver disease.

Objective To investigate the relationship of blood glucose to Child-Turcotte-Pugh scores in patients with alcoholic liver cirrhosis. Methods We included 76 patients with alcoholic liver cirrhosis in this study and their blood glucose levels and Child-Turcotte-Pugh scores were obtained. Results The blood glucose levels in Child-Turcotte-Pugh class A,B and C were 5.59±0.72mmol/L,5.45±0.85mmol/L and 7.04±2.82mmol/L, respectively;the blood glucose in class C was much higher than that in patients with class A or B; the blood glucose in patients with complications was much higher than that in patients without （9.24±2.53 mmol/L vs. 5.23±1.62 mmol/L,P<0.05）. Conclusion The patients with alcoholic liver cirrhosis are always complicated by glucose metabolic abnormality, which is sometimes correlated to the injuries of liver functions.

Objective To inhibit the gene expression of connective tissue growth factor （CGTF） by small interfering RNA（siRNA） and observe its influence on downstream gene expression. Methods 293T cells were cotransfected with lent viral transferred plasmid containing siRNA targeted to CTGF,packaging plasmid,and envelope protein plasmid. The lent viral vector was packed to mediate CTGF gene silence and infect hepatic stellate cell line HSC-T6 cells. The lent viral vector with the highest inhibition efficiency was screened by real-time PCR and Western blot. Efficiency of infection was measured by the expression of green fluorescent protein. mRNA levels of tissue inhibitor of metalloproteinase-1 （TIMP-1）,TIMP-2,and collagen I were detected by real-time PCR. Results Efficiency of infection of the packing lent viral vector was more than 50%. After infecting HSC-T6 cells,the expression of CTGF was suppressed on both mRNA and protein level,with the highest inhibition efficiency by TS1. The expressions of mRNA in TIMP-1,TIMP-2,and collagen I were significantly restrained by inhibition of CTGF（P<0.05）. Conclusion The packing lent viral vector in HSC-T6 has a better efficiency of infection and CTGF gene silencing effect. Gene silencing can further inhibit the TIMP-1,TIMP-2 and collagen type I gene expression in HSC-T6 cells. It has an active significance in anti-fibrosis studying.

Objective To study the distribution and expression of β-galactosidase（LacZ） reporter gene mediated by adenovirus vector in the liver of normal mice or mice with fulminant hepatitis, so as to evaluate the efficacy and safety of adenovirus as a gene therapy vector transduction system. Methods The normal Balb/cJ mice were randomly assigned to normal and fulminant hepatitis group. Each mouse was injected with adenovirus at 2×10⁸PFU by tail vein injection,then the normal mice serum was collected to detect ALT and TBil at 24h,48h,72h,96h,120h, and the seventh day,meanwhile,the mice organs including liver,lung,heart,and kidney were collected and stained by X-gal in order to observe the expression of adenovirus vector in mice organ;the expression of adenovirus vector in liver of fulminant hepatitis mice induced by MHV-3 was detected. Results Adenovirus mainly expressed in the livers of mice;In normal group and fulminant hepatitis group,the highest expression efficiency was at 72 hours,reached 55.7% and 25.7%,respectively,then it followed by a gradual reduction of adenovirus expression; The adenovirus infection didn’t show obvious injury in liver function. Conclusions The adenovirus vector is highly expressed in the liver with no significantly liver damage; Adenovirus vector might be as an effective and safe gene delivery system for liver disease gene therapy.

Objective To investigate the correlations of serum HBV DNA and HBsAg and HBeAg levels in patients with chronic hepatitis B. Methods 951 patients with CHB were enrolled. Serum HBV DNA was tested by fluorescence quantitative polymerase chain reaction,and HBsAg and HBeAg were detected by chemiluminesent microparticle immunoassay. Results 53.6% patients were HBV DNA positive in 951 patients with CHB; The serum HBsAg and HBeAg concentrations were positively correlated with serum HBV DNA levels（rs=0.45 and re=0.49, respectively,P<0.05）;The serum HBV DNA were divided into 3 groups（eg.<3 lgcopies/ml,3 to 7 lgcopies/ml and ≥7 lgcopies/ml）.There was an significant differences in serum HBsAg and HBeAg levels among the 3 groups （P<0.05）;There was also an significant differences in serum HBV DNA levels among different serum HBsAg levels（eg. <1000 IU/ml,1000~10000 IU/ml and ≥10000 IU/ml,P<0.05）. Conclusion Serum HBsAg concentration is related to serum HBV DNA levels in patients with hepatitis B,but it is not feasible in most cases to use HBsAg concentration for monitoring their serum HBV levels .

Objective To investigate the effect of telbivudine treatment in patients with HBeAg-positive hepatitis B. Methods 48 patients with HBeAg-positive hepatitis B were de novo treated with telbivudine. At 12， 24 and 48 weeks after treatment， the biochemical， virological and serological responses were evaluated. Results At 12，24 and 48 weeks after treatment， the serum response rates were 22.9%，56.2%，64.6%，respectively; the virological response rates were 47.9%，85.4%，89.6%，respectively. They were significantly correlated to baseline ALT，AST and HBV DNA levels; the biochemical response rates were 47.9%，83.3%，95.8%，respectively. Biochemical response at week 24 was correlated to baseline ALT and AST levels; serum creatine kinase levels were elevated to 192 to 610U/L in 3 patients without muscle symptomes. Conclusions Telbivudine has beneficial effects in chronic hepatitis B patients with HBeAg positive.

Objective To explore the relationship between liver fibrosis stages /inflammation grades and blood platelet counts and serum fibrosis indices in patients with chronic hepatitis B,and to find the better non-invasive indices for liver histological damage. Methods The liver fibrosis stages /inflammation grades in 136 CHB patients were confirmed by pathological examination using biopsy tissues. The blood platelet counts and serum fibrosis indices（pro-collagen III,collagen IV,hyaluronic acid,laminin） were tested in the patients. The correlation analysis was made between the aforementioned 5 indices and liver fibrosis stages /inflammation grades. Results Significant negative correlations were found between liver fibrosis stages /inflammation grades and blood platelet counts （Spearman r=-0.370 and -0.356,P=0.002 and 0.0003,respectively）; However,significant positive correlations were found between the liver fibrosis stages /inflammation grades and liver fibrosis indices（Spearman r for PC-III,C-IV,HA,and LN were 0.414,0.390,0.419,0.393,and 0.426,0.375,0.548,0.343,respectively,all P<0.05）. Conclusions The blood platelets,PC-III,C-IV,HA,and LN may be used as an noninvasive markers for liver injuries in CHB patients.

Objective To analyze the hepatic pathohistology of biopsies in chronic hepatitis B patients with mild ALT elevation. Methods 216 liver biopsies from patients with chronic hepatitis B with ALT elevation （lower than two times upper limits of normal） and 83 with asymptomatic HbsAg carriers （ASC） were included for liver biopsies. Results 55.6% and 62.1% of patients with CHB had G₂ or S₂ ,respectively,much higher than those in the ASC group（P<0.01）; The greater than G₂ or S₂ in patients with CHB over the age of 40 years were 69.5% or 75.0%,respectively,much higher than in patients younger than 40 years;In patients with chronic hepatitis B and with the age over 40 years,52.9% had G₃ to G₄ in those with ALT greater than 60u/L,much higher than 31.7% in patients with the ALT lower than 60U/L;The fibrosis above S₂ staging was 84.3% in patients with ALT greater than 60U/L,much higher than that in patients with ALT lower than 60U/L. Conclusion Part of patients with CHB with mild ALT elevation have liver pathological changes.The patients over the age of 40 years and ALT higher than 60U/L should receive antiviral treatment.

Objective To investigate the improvement of laboratory index by magnesium isoglycyrrhizinate （MIG） in patients with chronic liver diseases. Methods 66 patients with chronic liver diseases were treated with MIG and 129 with non-glycyrrhizic acids（non-MIG）,such as with diisopropylamine dichloroacetate alone or combined with one or two kinds of hepatoprotective agents. Clinical data and laboratory results were analysed,such as the changes of blood hematology,electrolytes,coagulation and liver biochemistry for two weeks. Results Compared with non-MIG,MIG was mainly used in DILI patients（42.4% vs 5.4%,P<0.01）;In MIG group,the level of the white blood cell（P=0.01） and platelet（P<0.01） elevated after the first week of the treatment,and the liver enzyme （ALT,AST,ALP,and GGT） decreased significantly（P<0.01）;At the initiation of the treatment,the levels of ALT,AST,GGT and ALP in MIG therapy group were 5-fold （P<0.01）,2.94-fold （P<0.01）,2.41-fold（P<0.01） and 1.63-fold（P<0.01） higher than non-MIG group,respectively. After the two-week-treatment,they decreased to 1.53-fold（P=0.03）,1.04-fold （P=0.79）,1.29-fold （P=0.26） and 1.1-fold （P=0.33）,respectively;There were almost no effects on electrolytes and coagulation indexes in MIG group（P>0.05,respectively）;After two week treatment,there were no significant differences between TP,ALB,and Hb in the two groups（P>0.05,respectively）. Conclusion MIG treatment is safe and effective in patients with chronic liver injury. Compared to the control group,enzymatic indicators in MIG group shows a quick recovery and no significant side effects.

Objective To investigate the presence of AMA-M₂ antibodies in 1039 patients with cryptogenic hepatitis. Methods M₂ antibodies were tested by ELISA. Results M₂ antibodies were detected in 81（7.9%）of the 1039 patients;Of the 81 cases,57 were female and 24 were male,and the mean age was 53.0 ± 14.8 year old,and 78 patients were diagnosed as PBC;The most common symptoms were fatigue（74.4%,58/78）,jaundice（61.5%,48/78）and skin itching（37.2%,29/78）;Besides,39（50.0%） patients were with ANA positive,and 14（18.0%） with anti-SSA/Ro positive. Conclusion On the diagnosis of the disease,the detection of serum antimitochondrial autoantibodies（AMA） is an important indicator. The detection of AMA-M₂ is important for early diagnosis of patients with PBC.

Objective To study the clinical effect of plasma exchange on liver failure. Methods Retrospective analysis was made on the clinical data in 216 patients with liver failure.According to the way of therapy,the 216 patients were divided into PE treatment group and control group. The changes of clinical symptom,biochemistry index,adverse reactions and prognosis were observed. Results Serum ALT,AST,TBIL and Cr levels in PE group decreased（394.20±156.40u/L vs. 144.15±34.84u/L,374.54±151.34u/Lvs. 253.47±106.74u/L,399.01±146.18μmol/L vs. 359.70±110.69μmol/L,189.72±121.94μmol/L vs. 139.14±82.53μmol/L,respectively,P<0.01）,serum CHE and PTA increased（2964.37±1475.45u/L vs. 4480.97±1009.84u/L,40.01±17.99% vs.49.49±23.46%,P<0.01）,while they didn’t changed in control group（P>0.05）;The improvement rate of patients in treatment group was better than in control group（61.86% vs. 34.70%,x²=15.63,P<0.001）,but the mortality rate in the PE group was lower than in the control（5.93% vs. 8.16%,x²=14.37,<0.001）; the incidence of complications such as hepatic encephalopathy and hepatorenal syndrome in dead patients were higher than in the survivals（32.5% vs. 10.0%and 20.0% vs. 0.0%,P<0.05）. Conclution Therapeutic plasma exchange significantly increase the improvement rate of patients with liver failure.

Objective To investigate the frequencies of regulatory T cells（Treg cells）,its subgroups and serum levels of IL-1β,IL-6 and IL-10 in HBV-related liver cirrhosis. Methods In 20 healthy persons,17 patients with chronic hepatitis B（CHB） and 38 with HBV-related liver cirrhosis（LC）,the frequencies of CD4⁺CD25⁺Fox3⁺ cells （Treg cells）,CD4⁺CD39⁺Fox3⁺cells（CD39⁺Treg cells）,and CD4⁺CTLA-4⁺ Fox3⁺cells（CTLA-4⁺ Treg cells） were detected by flow cytometry. The serum levels of IL-1β, IL-6 and IL-10 were detected by ELISA. Results The frequencies of Treg cells,CD39⁺ Treg cells and CTLA-4⁺Treg cells and the serum levels of IL-1β, IL-6 and IL-10 in patients with LC were higher than that in normal persons（P<0.01） and in with CHB（P<0.05 or P<0.01）;the frequencies of Treg cells,CD39⁺Treg cells and serum levels of IL-6 in patients with decompensated LC were higher than that in with compensated cirrhosis（P<0.01）; in LC group,the frequencies of Treg cells were positively correlated with CD39⁺ Treg cells（r=0.474,P<0.01）and CTLA-4⁺Treg cells（r=0.330,P<0.05）;the frequencies of Treg cells（r=0.381,P<0.05） and CD39⁺ Treg cells （r=0.333,P<0.05） were positively correlated with Tbil. The frequencies of Treg cells（r=-0.549,P<0.01）,CD39⁺ Treg cells（r=-0.437,P<0.01） and CTLA-4⁺Treg（r=-0.368,P<0.01） cells were negatively correlated with PTA,and the frequencies of CD39⁺Treg cells were positively correlated with AST（r=0.406,P<0.05）. Conclusion Changes of frequencies of Treg cells, its subgroups and levels of IL-1β,IL-6,IL-10 can be as a auxiliary examination to give a clinical diagnosis of HBV-related liver cirrhosis who are about to receive initial anti-viral treatment.

Objective To observe the clinic effect of bone marrow stem cell transplantation by hepatic artery on patients with decompensated cirrhosis so as to find a new approach and a new methods to the treatment of severe liver disease. Methods Totally 87 patients with decompensated cirrhosis from our hospital，aged 27 to 65 year old（the average age was 46.7±8.3 year old） were enrolled. Among them，sixty-seven were hepatitis B cirrhosis，two were hepatitis C cirrhosis，eight were primary biliary cirrhosis，nine were alcoholic cirrhosis，and one was cirrhosis of unknown cause.Under sterile condition，200ml of bone marrow was harvested from eighty-seven patients with decompensated cirrhosis，and then BMSCs were isolated and purified. Under local anesthesia，isolated BMSCs were injected into the livers of the patients via hepatic artery.The clinical efficacy，improvement of symptoms and adverse events were observed.　Results Four weeks after transplantation，the levels of serum albumin increased from 31.5±5.1g /L to 33.9 ± 6.0g/L（P<0.01）；the levels of prothrombin time decreased from 17.4±5.6s to 15.8±4.7s（P<0.01）；there were no differences about the levels of aspartate aminotransferase，total bilirubin and alpha-fetoprotein（P>0.05）；pleural effusion disappeared in 10 cases，ascites disappeared in 23 cases，abdominal distension alleviated in 17 cases，and lower extremity edema subsided in 21 cases. No serious complications occurred in this series. Conclusion The bone marrow stem cell transplantation by hepatic artery in treatment of patients with decompensated cirrhosis has a good short-term effects without side effects. It should be worthy of further investigation.

Objective To evaluate the efficacy of sclerotherapy plus octreotide for esophageal variceal hemorrhage. Methods We searched CBM,the Cochrane Library,the Cochrane Controlled Trials Register,MEDLINE,EMBASE and SCIE for randomized controlled trials about sclerotherapy plus octreotide versus sclerotherapy only. The Meta-analyses were conducted by using The Cochrane Collaboration’s RevMan 4.2 software. Results Five randomized controlled trials involving 511 patients were included including 255 patients in sclerotherapy plus octreotide and 256 patients in sclerotherapy alone group. The rates of early hemostasis,the rates of rebleeding,the rates of mortality and adverse events were analyzed. Meta-analyses showed that the early hemostasis in sclerotherapy plus octreotide group was better than in sclerotherapy group（OR=2.65,95%CI=1.66 to 4.23）;rebleeding was more frequent in sclerotherapy group（OR= 0.52,95%CI=0.32 to 0.84）;However,the incidence of death was similar in both groups（OR=0.75,95%CI=0.44 to 1.29）;The most frequent adverse events was hyperglycemia,esophageal ulcer,hepatic encephalopathy and headache,and it showed no significant difference in the two groups. Funnel plot showed symmetrical,suggesting the lover possibility of publication bias. Conclusions Sclerotherapy plus octreotide therapy is better for hemostasis of esophageal variceal bleeding than sclerotherapy alone.