12 Liver and Biliary System

12 Liver and Biliary System

2015091 Biological function and molecular mechanism of URI in HepG2 cells.ZHOUWei（周?chē)?，et al. Dept Cell&Molecular Biol，Med Coll，Soochow Univ，Suzhou 215123.Chin JOncol2014；36（11）：816-824.

ObjectiveTo explore the effect themolecularmechanism of the unconventional prefoldin RPB5 interactor（URI）in hepatocellular carcinoma HepG2 cells.M ethodsThe cDNA sequence and shRNA of URIwere obtained and sub-cloned into eukaryotic expression vectors. Then those vectors were transfected into HepG2 cells to obtain stable transfection cell line.The cell proliferation and anchor-independent growth in URI-overexpressing and knockdown HepG2 cells were determined by CCK-8 and soft agar colony assay.Flow cytometry was applied to detect the cell cycle and apoptosis ofγ-ray irradiated cells.Apoptosis related geneswere detected by Western blot.ResultsThe pCDNA3.1-URI and pGPU6-URIi eukaryotic expression vectors were constructed successfully and corresponding stable transfection cell lines were obtained.Cell proliferation rates of the HepG2，pCDNA3.1-URI-HepG2 and pGPU6-URIi-HepG2 cellswere（588.78±32.12）%，（959.33±58.8）%and（393.93±39.7）%，respectively（P＜0.05）.The number of cell clones of HepG2，pCDNA3.1-URIHepG2 and pGPU6-URIi-HepG2 cellswere43±7，85± 5 and 20±4（P＜0.05），respectively.Afterγ-ray irradiation，the URI-overexpressing cell line showed a significantly lower apoptosis rate and G2/M phase arrest than those in the URI-depleted cell line（P＜0.05）.In the HepG2 cells，the relative protein expression levels of URI，Box and Bcl-2 were 0.92±0.03，1.11±0.13 and 0.82±0.01（P＜0.05）.In the pCDNA3.1-URI± HepG2 cells，the relative protein expression levels of URI，Bax and Bcl-2 were 1.79±0.12，0.48±0.01 and 2.20±0.30（P＜0.05），respectively.In the pGPU6-URIi-HepG2 cells，the relative protein expression levels of URI，Bax and Bcl-2 were0.50±0.04，1.52± 0.20 and 0.38±0.01（P＜0.05），respectively.The expression of Bax was down-regulated and Bcl-2 was upregulated in the URI-overexpressing cell line.However，on the contrary，expression of Bax was up-regulated and Bcl-2 was down-regulated in the URI-depleted cell line.ConclusionURImay promote the growth of hepatocellular carcinoma cells via inhibition of cell proliferation and reducing the apoptosis in hepatocellular carcinoma cells in vitro.After the impairment of URI expression，the proliferation ability of hepatocellular carcinoma cells is suppressed and the ability to resistγ-ray irradiation is reduced.URImay become a potential new target for cancer therapy of hepatocellular carcinoma.

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2015092 Imaging characteristics of hepatobiliary scintigraphy in neonatal intrahepatic cholestasis caused by citrin deficiency.ZHANG Meihong（張梅虹），et al.Dept Pediatr，Jinshan Hosp，F(xiàn)udan Univ，Shanghai201508.Chin JNucl Med Mol Imaging 2014；34（5）：358-361.

Ob jectiveTo investigate the scintigraphic features of neonatal intrahepatic cholestasis caused by citrin deficiency（NICCD）and to explore the clinical significance of99Tcm-EHIDA hepatobiliary scintigraphy.M ethodsHepatobiliary scintigraphy with99Tcm-EHIDA was performed in 28 genetic confirmed NICCD cases（16 males，12 females，1-8 months）.Normal dynamics（i.e.，time-activity curve）of the tracer in heart，liver and kidneys was defined as normal hepatic uptake function.Decreased liver uptake of the tracer and/or prolonged heart and kidneys radioactivity retention was defined as impaired hepatic uptake function.Normal or delayed biliary excretion was defined as radioactivity appearing in the bowelwithin 60 min or after 60 min.Occluded biliary excretion was defined as absentbowel radioactivitywithin 24 h.The characteristics of hepatobiliary scintigraphy and their relationship with levels of serum total bilirubin（TB），direct bilirubin（DB），ALT，total bile acid（TBA）were retrospectively analyzed.Statistical analysis was performed using Kruskal-Wallis rank sum test.Resu ltsOf28 NICCD patients，20 showed normal uptake while 8 had impaired hepatic uptake.Ten of twenty patientswith normal uptake function showed normal biliary excretion and the others showed delayed biliary excretion.Four of eight cases who presented impaired hepatic uptake function were with delayed biliary and the rest displayed biliary excretion occlusion.Compared with the patients with normal hepatic uptake function，patients with impaired hepatic uptake had significantly higher levels of TB and DB［TB：183.6（128.7-280.9）mmol/L vs105.5（80.0-141.7）mmol/L，Z=-2.25；DB：135.6（95.7-212.6）mmol/L vs 73.1（53.9-97.9）mmol/L，Z=-2.73；both P＜0.05］.Compared with those with normalbiliary excretion function，patientswithdelayed biliary excretion had significantly higher levels of TB，DB and TBA［TB：137.5（122.0-170.9）mmol/L vs81.7（65.7-93.5）mmol/L，Z=-3.92；DB：96.5（81.1-108.0）mmol/L vs54.1（45.3-72.6）mmol/ L，Z=-3.74；TBA：245.6（183.9-299.2）mmol/L vs136.0（73.5-163.2）mmol/L，Z=-2.57；all P＜0.05］.The levels of TB［262.0（152.1-542.8）mmol/L］and DB［192.7（118.1-407.2）mmol/L；both Z=-2.82；both P＜0.05］were the highest in patients with occluded biliary excretion.Compared with the patients with delayed excretion，the occluded excretion subgroup had significantly higher levels of ALT［71.5（48.5-144.8）U//L vs20.0（16.5-27.7）U/ L，Z=-2.66，P＜0.05］.Conclusion99Tcm-EHIDA hepatobiliary scintigraphy may evaluate hepatic uptake and excretion function of the NICCD infants.When the hepatic uptake is remarkably decreased，the occluded biliary excretion can be shown.

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2015093 Efficacy and safety of telbivudine for pregnant wom en w ith hepatitis B e antigen negative chronic hepatitis B.YUE Xin（岳欣），et al.Dept Gynaecol &Obstetrics，2nd Hosp，Nanjing 210003.Chin JInfect Dis 2014；32（9）：550-553.

Ob jectiveTo evaluate the efficacy and safety of telbivudine for pregnant women with hepatitis B e antigen（HBeAg）negative chronic hepatitis B（CHB）.MethodsSixty-two cases of HBeAg negative CHB pregnantwomen were collected from May 2007 to May 2012，and they were divided into telbivudine group（n=31，600 mg per day by oral administration）and compound glycyrrhizin group（n=31，120 mg per day by intravenous administration）.All neonateswere given intramuscular injection of 200 IU hepatitis B immune globalin at birth immediately and 15 days after birth，and 20μg genetically engineered hepatitis B vaccine at 0，1 and 6 months after birth.The serum alanine aminotransferase（ALT）level and hepatitis B virus（HBV）DNA titer were monitored. The HBV DNA negative conversion rate，the rate of intrauterine infection，duration of pregnancy，delivery mode，neonate weight and disability rate were compared between groups.All categorical data were analyzed using the chi-square test and comparison between groups was analyzed by t test.ResultsIn telbivudine group，the HBV DNA level before delivery［（0.20±0.11）lg copy/mL］and 6 weeks after delivery［（0.22±0.13）lg copy/mL］were lower than that before treatment［（6.24±0.75）lg copy/mL］and the differences were statistically significant（t=303.128 and 301.321，respectively；both P＜0.01）.The negative conversion rate of HBV DNA in telbivudine group was 28 cases before delivery，while in compound glycyrrhizin group，no one had HBV DNA negative conversion.And statistical significant differences were achieved between these two groups before delivery and 6 weeks after delivery（t= -20.285 and-8.721，respectively；both P＜0.01）. In telbivudine group，the ALT levels before delivery and 6 weeks after delivery were（13.08±5.87）U/L and（25.97±17.48）U/L，respectively，which were significantly decreased compared with that before treatment（205.95±95.69）U/L.The differences were statistically significant（t=93.128 and 81.321，respectively；both P＜0.01）.In compound glycyrrhizin group，the ALT level before delivery［（104.15±69.15）U/L］was lower than that before treatment［（209.60±102.24）U/L］and the difference was statistically significant（t= 9.281，P=0.032）.However，the ALT level was fluctuant 6 weeks after delivery（150.26±86.43）U/L，which was not significantly different from that before treatment（t=2.821，P=0.122）.The ALT levels before delivery and 6 month after delivery were significantly different in both two groups（t=-2.559 and-3.158，respectively；both P＜0.05）.Therewere no statistically significant differences between these two groups in the rate of intrauterine infection，duration of pregnancy，delivery mode，neonate weight and disability rate.ConclusionThe using of telbivudine for pregnantwomen with HBeAg negative CHB can effectively control the hepatitis activation and reduce the virus titer.

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2015094 Association between apolipoprotein B/A-I and non-alcoholic fatty liver disease.LüXiaofei（呂曉斐），et al.Dept Endocrinol Metab，Ruijing Hosp，Shanghai Jiaotong Univ Med Sch，Shanghai 200025. Chin JEndocrinol Metab 2014；30（10）：826-829.

ObjectiveTo study the association between apoli-poprotein B/A-I（ApoB/A-I）and the risk of prevalence of non-alcoholic fatty liver disease（NAFLD）.M ethodsThis was a community-based cross-sectional analysis conducted among2 519 residents aged 40 years or older from Jiading District，Shanghai，China.All subjects underwent a standard questionnaire，anthropometric measurements，and biochemical evaluation.Non-alcoholic fatty liver disease was defined by hepatic ultrasonography.After excluding subjects with excess alcohol consumption and medical history of liver diseases，more than three times of normal serum alanine aminotransferase（ALT），aspartate aminotransferase（AST），andγ-glutamyltransferase（CCT）levelswere obtained in 2 139 subjects.Unadjusted and multivariate adjusted logistic regression analyses were applied to estimate the association between ApoB/A-I ratio and the risk of prevalent NAFLD.ResultsIncreased ApoB/A-I ratiowas associated with a less favorable metabolic profile.The prevalence of NAFLD increased in accordance with the quartiles of ApoB/A-I ratio（14.3%，23.6%，30.4%，40.4%，respectively，P＜0.01）.In the multivariate adjusted logistic regression analysis，the risk of having NAFLD increased by 105%with each unit increment in ApoB/A-I ratio（OR=2.05，95%CI 1.19-3.53.P=0.01）.ConclusionApoB/A-I ratio is associated with the risk of prevalent NAFLD，being independent of other established metabolic risk factors.

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2015095 The exp ression of circu lating m icroRNA-595 in patients w ith hepatitis B virus-related liver failure and its relationship with disease prognosis.WANG Peng（王芃），et al.Dept Infect Dis，Ruijin Hosp，Shanghai Jiaotong Univ Med Sch，Shanghai 200025.Chin J Infect Dis 2014；32（9）：545-549.

ObjectiveTo measure the expression of circulating microRNA（miRNA）in patients with hepatitis B virus（HBV）-related liver failure and its relationship with disease prognosis.M ethodsThemiRNA expressions in serum of 5 patients with HBV-related liver failure and 5 healthy control subjects were compared using Exiqon miRCURY LNATMmiRNA microarray.The sera from 20 patients with chronic hepatitis B（CHB），20 patients hepatitis B related cirrhosis，50 patientswith HBV-related liver failure and 40 healthy persons in Ruijin Hospital were collected.The relative expression of miRNA-595 was measured using quantitative real-time polymerase chain reaction（PCR）.The relative expressions ofmiRNAs among groups were analyzed using student t test，the correlations were analyzed by Pearson and Spearman correlation.Resu ltsMicroarray informed that92 miRNAs changed significantly in patients with HBV-related liver failure，and miRNA-595 increased most significantly. The results of real-time PCR showed that the relative expressions of miRNA-595，miRNA-300 and miRNA-122 were 6.03（t=3.134，P=0.003），3.12（t=7.221，P＜0.01）and 2.77（t=2.671，P=0.021），which were higher compared to those in healthy control group. In the analysis of the relationship between miRNA-595 expression and disease prognosis in patients with HBV-related liver failure，the relative expressions of miRNA-595 in patients with CHB，hepatitis B related cirrhosis and HBV-related liver failure were 2.26（t=3.780，P= 0.001），3.32（t=6.111，P＜0.01）and 6.03（t= 3.134，P=0.003），respectively，which were all increased compared to that of the healthy control.The relative expression ofmiRNA-595 of patientswith HBV-related liver failure was2.66 times（t=2.450，P=0.043）higher than that of patients with CHB.When dividing patients according to prothrombin activity，miRNA-595 increased significantly in patients with early stage liver failure.When dividing patients according to model of end-stage liver disease（MELD）score，MELD score was positive correlated with the expression of miRNA-595 when MELD score was under 30（r=0.673，P= 0.004）.The expression of serum miRNA-595 in survival group（11.08，n=23）was higher than that in non-survival group（3.67，n=27，t=4.309，P=0.041）.ConclusionThe expressions ofmiRNA595，miRNA-300 and miRNA-122 are all increased in patientswith HBV-related liver failure，especially the expression of circulatingmiRNA-595 at early stage of the disease.The miRNA-595 may be used as a new serum biomarker formonitoring the severity of disease.

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2015096 Analysis of epidem iological and clinical features of 621 patientswith acute hepatitis E.LIAOBaolin（廖寶林），etal.Dept Infect Dis，Guangzhou 8th People's Hosp，Guangzhou 510060.Chin J Infect Dis 2014；32（9）：554-559.

ObjectiveTo investigate the epidemiological and clinical features of acute hepatitis E（AHE）.MethodsAll the data of AHE patients from April 2005 to October 2011 were collected and their epidemiological features were retrospectively analyzed.Patients were divided into two groups：patientswith single hepatitis E virus（HEV）infection and patientswith HEV/hepatitis B virus（HBV）coinfection，to compare the biochemical parameters and outcomes and to find out the risk factors for AHE related liver failure.Kruskal-Wallis test，Chi square test，and Logistical regression analysis were used for statistical analysis.ResultsA total of 621 cases were included in the present study and most patients were elderly male mostly from February to May every year.The incidence of AHE related liver failure and mortality was 18.68% and 1.93%，respectively.Compared to the single HEV group（n=331），the HEV/HBV group（n=280）had a longer hospital stay（46 d vs 40 d，Z=-4.591，P＜0.01），a significantly lower prothrombin activity（55.5%vs 78.7%，Z=-7.998，P＜0.01）and a significantly higher incidence of AHE related liver failure（30.7%vs 9.1%，χ2=46.229，P＜0.01）.In single HEV related liver failure group（n=30），the percentages of early-stage，interim-stage and end-stage live failure were53.33%，23.33%and 23.33%，respectively. While in the HEV/HBV related liver failure group（n= 86），the corresponding numbers were 34.88%，31.40%and 33.72%，respectively.The differences were not statistically significant（χ2=3.176，P= 0.204）.Additionally，the clinical outcome between these two groups was also comparable（83.33%vs 91.86%，χ2=0.945，P=0.331）.The Logistic analysis showed that being over 50 years（OR=2.080，P= 0.002）and coinfection with HBV（OR=5.632，P＜0.01）were risk factors for AHE related liver failure.ConclusionAHE is seasonal and mainly occurs in elderlymale.Advanced age and HBV coinfectionmay be risk factors of severe AHE.

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2015097 Hepatitis B virus-3p-siRNA inhibits hepati-tis B virus replication and activates interferon-βexp ression in m ice.XING Yaling（邢雅玲），et al.Instit Radiat Med，MilitMed Sci Acad，Beijing 100850.Chin J Infect Dis2014；32（9）：517-521.

ObjectiveTo observe the activation of anti-viral innate immune response of typeⅠinterferon and inhibition of hepatitis B virus（HBV）genome replication inmice by HBV-3p-siRNA.MethodsHBV-3p-siRNA was designed by targeting specific sequence of HBV S/P mRNA and was generated by in vitro transcription.Negative control siRNA（NC-siRNA）and non-modified HBV-siRNA were used as control groups.Blood samples were collected from tail vein of mice and the model of HBV-infected mice were established by hydrodynamic injection.Forty mice were divided into 4 groups with 10 in each group. Themodel group was only injected with pGL3.0-HBV1. 2 copy plasmid.The negative control group received peritoneal injection of NC-siRNA.HBV-siRNA group received peritoneal injection of HBV-siRNA and HBV-3psiRNA group received peritoneal injection of HBV-3psiRNA.The interferon-β（IFN-β）and hepatitis B surface antigen（HBsAg）in serum were detected by enzyme linked immunosorbent assay（ELISA）.The copies of HBV DNA were assessed by fluorescence quantitative polymerase chain reaction（PCR）.The statistical difference between groups was determined using One way-ANOVA analysis by LSD or Dunnett T3.Resu ltsSerum level of IFN-βwas（12.37±5.32）pg/mL in model group，（22.61±6.29）pg/mL in negative control group，（26.40±5.39）pg/m L in HBV-siRNA group and（68.37±21.00）pg/m L in HBV-3p-siRNA group. The secretions of IFN-βinto serum were significantly enhanced by HBV-siRNA and HBV-3p-siRNA compared with model group（F=23.988 and 46.523，respectively，both P＜0.01）.Serum level of HBsAg was（2 864.86±907.11）ng/mL in model group，（2 198.86±456.89）ng/mL in negative control group，（1 049.71±396.28）ng/mL in HBV-siRNA group and（640.86±383.08）ng/m L in HBV-3p-siRNA group. The expressions of HBsAg were inhibited by HBV-3psiRNA and HBV-siRNA compared with model group（F=23.537 and 39.144，respectively；P=0.025 and 0.010，respectively）.Serum level of HBV DNA was（2.54×104±1.46±104）copy/mL in model group，（2.22×104±2.62×103）copy/mL in negative control group，（3.59×103±2.88×103）copy/m L in HBV-siRNA group and（2.65×103±1.46×103）copy/mL in HBV-3p-siRNA group.Serum level of HBV DNA were inhibited by HBV-3p-siRNA and HBV-siRNA compared with model group（F=15.013 and 16.741，respectively，both P＜0.05）.All of the indicated siRNA used in the experiments showed no apparent effects on the body mass index of the mice models.ConclusionHBV-3p-siRNA，which induces the production of IFN-β and inhibits HBV replication through gene silencing in vivo，may be a powerful bifunctional antiviralmolecule.

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