Expression and clinical significance of metastasis-associated in colon cancer 1 (MACC1) in HBV-related hepatocellular carcinoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To clarify the expression and clinical significance of metastasis-associated in colon cancer 1 (MACC1) mRNA in hepatocellular carcinoma (HCC).</p><p><b>METHODS</b>The expression and distribution of MACC1 were assessed by quantitative real-time polymerase chain reaction (RT-PCR) and immunohistochemical staining (IHC) in a cohort of hepatitis B virus-related HCC, including 138 in early (A), 96 in intermediate (B) and 120 in advanced stages (C). The association of MACC1 mRNA with disease progression and outcomes was analyzed by univariate and multivariate Cox analysis.</p><p><b>RESULTS</b>The intratumoral expressions of MACC1 mRNA in HCC stage I (0.001 76, range: 0.000 54 - 0.002 47), stage II (0.002 49, range: 0.000 55 - 0.006 78) and stage III (0.008 35, range: 0.006 86 - 0.009 88) were about 3-, 4- and 14-fold higher than that in the normal liver tissue (0.000 59, range: 0.000 57 - 0.000 60), respectively. Intratumoral expression of MACC1 mRNA increased with disease progression from stage I to stage III. HCC clinical staging classification, age, portal vein invasion and tumor differentiation were significantly associated with intratumoral high expression of MACC1 mRNA (All P < 0.05). Immunohistochemical staining showed that there was an increased MACC1 expression in cytoplasm of HCC cells and positive nuclear staining in some cases. Increased MACC1 mRNA expression could predict poor outcome and recurrence in stage A and B HCC postoperatively. The median tumor-free survival and total survival of patients with high MACC1 mRNA expression were 34.0 and 40 months, respectively, significantly lower than that in those with low expression (48.0 and 48.0 months) (all P < 0.01). Cox analysis showed that Child-Pugh grading and high expression of MACC1 mRNA were independent predictive factors, and high expression of MACC1 was an independent predictive factor affecting the tumor-free survival.</p><p><b>CONCLUSIONS</b>MACC1 mRNA up-regulation is a feature of disease progression in HCC. MACC1 mRNA expression in the HCC may become an independent predictive factor for recurrence and survival in postoperative HCC patients.</p>

Clinical efficacy and prognostic factors for cryoablation patients with advanced hepatocellular carcinoma / 中华肝脏病杂志

<p><b>OBJECTIVES</b>Investigate the clinical efficacy of cryotherapy ablation treatment for advanced hepatocellular carcinoma. analyse the predictive factors of cryotherapy ablation treatment.</p><p><b>METHODS</b>There were 190 cases of hepatitis B-related HCC patients with advanced HCC from 2005 to 2008 in our hospital. By using clinical cohort method, they included cryoablation group (147 cases) and control group (43 cases), The median survival time and time to disease progression were compared. Evaluate clinical significance of age, gender, location of portal vein tumor thrombus, HBeAg, tumor histological grade, Child-Pugh classification, end-stage liver disease (MELD) score, advanced liver cancer prediction system (ALCPS) score and the Eastern Cooperative Oncology Group performance status (ECOG PS) score for predicting the efficacy of cryoablation. Group rates were compared with the x2 test, survival analysis by using Kaplan-Meier method, survival rates were compared by Log-rank analysis; multiple factor survival analysis by using Cox regression model.</p><p><b>RESULTS</b>Median survival time of cryoablation group and Control group was 7.5 (4.2 to 14.6) months and 3.2 (1.2 to 8.6) months, median TTP was 3.5 (2.5 to 4.5) months and 1.5 (1.0 to 3.5 months), the differences were statistically significant ( P less than 0.05 ). Median OS and TTP of advanced HCC patients who had Well-differentiated tumor, Child-pugh A-class and low score of MELD score, ALCPS score, ECOG PS score were significantly longer than the poorly differentiate, Child-Pugh B-class and the high those scores ( P less than 0.05). ECOG PS ( P less than 0.05, 95% CI 1.074 to 2.143) and ALCPS (P less than 0.05, 95% CI 1.005 to 2.121) were independent predictors for OS of advanced HCC.</p><p><b>CONCLUSION</b>Cryoablation treatment can prolong median OS and TTP of advanced HCC; ECOG PS and ALCPS are important predictors for survival time of advanced HCC.</p>

Liver microvessel density in evaluating therapeutic effect of sorafenib combined with argon-helium cryoablation in treatment of advanced hepatocellular carcinoma / 第二军医大学学报

Objective To investigate the therapeutic effect of argon-helium cryoablation combined with sorafenib on hepatitis B-related advanced hepatocellular carcinoma (HCC) and to evaluate the role of microvessel density (MVD) in prognostic evaluation. Methods A total of 102 patients with advanced HCC were randomly divided into two groups, with 50 receiving sorafenib plus cryoablation(group S&-C) and 52 receiving only cryoablation (group C). The endpoint of the treatment was tumor progression or untolerable adverse reaction. Tumor tissues were obtained before treatment. MVD was evaluated by immunohistochemical analysis using CD34 antibody. The therapeutic effects were evaluated according to RECIST criterion every 4-6 weeks. The adverse events were observed in both groups; the therapeutic effect, overall survival time (OS), and time to progression(TTP) were compared between two groups. Results In group ScV-C, complete response (CR) was achieved in 2 patients(4%), partial response (PR) in 9 (18%), and stable disease(SD) in 22(44%), with the disease control rate(DCR) being 66%; in group C. no CR, PR in 4(7. 6%), and SD in 19(36. 5%), with the DCR being 44. 2% (P<0. 05). The overall survival (OS) and the time to progression (TTP) were significantly longer in group S&-C than in group C (12. 5 months vs 8. 6 months, 9. 6 months vs 5. 3 months; P<0.01). The mean MVD in group CR&.PR (111/0.74 mm2) was significantly lower than that in group PD( 339/0. 74 mm2, P<0.01). Patients with lower MVD receiving sorafenib plus cryoablation had longer OS and TTP than those only receiving cryoablation. The OS and TTP in patients with higher MVD had no significant difference between different groups. Conclusion Sorafenib plus cryoablation is a safe and effective therapy for patients with advanced HCC; it can improve the OS and TTP of patients. Patients with higher MVD have a lower response to therapy and poor prognosis.

Mutual interaction between hepatitis B virus antigen and metallothionein / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To screen and identify the protein interacting with HBV antigen in hepatocytes. Then investigate the biological functions of hepatitis B virus antigen in the pathogenesis of hepatitis B and seek effective methods to prevent and treat it.</p><p><b>METHODS</b>The yeast two-hybrid system-3 technique was used to construct HBV PreS2, HBeAg, HBcAg, HBxAg bait plasmids. The bait plasmids transformed the yeast AH109 and expressed themselves in it. After being identified by SDS-PAGE and Western blot, the AH109 yeast was mated with yeast Y187 containing liver cDNA library plasmid in 2 x YPDA medium to form diploid yeast and was then plated on synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) and synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) containing x-alpha-gal for screening. Plasmids of blue colonies were extracted and transformed into Escherichia coli, then analyzed by DNA sequencing and bioinformatics. To further prove the interaction between HBV antigen and metallothionein, translation was performed by using reticulocyte lysate and coimmunoprecipitation was displayed in vitro.</p><p><b>RESULTS</b>Genes coding for HBV antigen binding protein were successfully cloned and metallothionein was found in that protein. The interaction between HBeAg, HBcAg and HBxAg and metallothionein were further proved by coimmunoprecipitation in vitro.</p><p><b>CONCLUSION</b>The interaction between HBV antigen and metallothionein indicates that metallothionein may participate in the pathogenesis of hepatitis B</p>

Cloning of genes transactivated by hepatitis B virus X protein / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To construct a subtractive cDNA library of genes transactivated by hepatitis B virus X protein (HBX) using suppression subtractive hybridization (SSH) technique and to clone genes associated with HBX transactivating function.</p><p><b>METHODS</b>The mRNA was isolated from HepG2 cells transfected with pcDNA3.1(-)-X and pcDNA3.1(-) empty vector respectively, then cDNA was synthesized. After restriction enzyme RsaI digestion, a number of small size cDNA was obtained. Then tester cDNA was subdivided into two portions and each was ligated with different cDNA adaptor. After tester cDNA was hybridized with driver cDNA twice and underwent nested polymerase chain reaction (PCR) twice the production was subcloned into T/A plasmid vectors to set up the subtractive cDNA library. Amplification of the library was carried out with E. coli strain JM109, some cDNA was sequenced and analyzed in GenBank with Blast.</p><p><b>RESULTS</b>The subtractive cDNA library of genes transactivated by HBX was constructed. The amplified library contained 85 positive clones, and colony PCR showed that these clones contained 200-1000 bp inserts. 65 clones were analyzed by sequencing and bioinformatics, which suggested nineteen known genes and fifteen genes with unknown function.</p><p><b>CONCLUSION</b>A subtractive cDNA library of genes transactivated by HBX using SSH technique has been constructed successfully, which may bring some new clues for studying the biological functions of HBX and the pathogenesis of hepatoma.</p>

Screening of the genes of hepatitis B virus PreS2 interacting proteins / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To screen and clone the genes of proteins in hepatocytes interacting with hepatitis B virus (HBV) PreS2 by yeast-two hybridization technique.</p><p><b>METHODS</b>The HBV PreS2 gene was amplified by polymerase chain reaction (PCR) and HBV PreS2 bait plasmid was constructed by using yeast-two hybridization system 3, then transformed into yeast AH109, followed by mating with yeast Y187 containing liver cDNA library plasmid in 2 YPDA medium. Diploid yeast was plated on synthetic dropout nutrient medium (SD/-Trp-Leu-Ade-His) and synthetic dropout nutrient medium (SD/-Trp-Leu-Ade-His) containing X-alpha-gal for selecting positive blue clones, then amplified by PCR, sequenced, and performed bioinformatics analysis.</p><p><b>RESULTS</b>HBV PreS2 gene was cloned successfully and expressed in yeast AH109.Twenty-six positive colonies were selected, among them, twelve containing metallothionein 2A, one cytochrome C oxidase II, two cytochrome P450 subfamily IV4F, two cytochrome c oxidase subunit 4 isoform 1, three albumin (ALB), one Na(+)K(+) transporting ATPase beta-1 polypeptide, two prealbumin, one lectin galactoside-binding subunit, and Two new genes with unknown function.</p><p><b>CONCLUSION</b>Genes of HBV PreS2 interacting proteins have been successfully cloned, which brings some new clues for studying the biological functions of HBV PreS2 and related proteins.</p>

Effect of HanDanGanLe on the cytokines in fibrotic rats / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To study the expression of connective tissue growth factor (CTGF) mRNA and transforming growth factor beta 1 (TGFbeta1) mRNA in immunity-induced liver fibrosis rats and the effect of HanDanGanLe on them.</p><p><b>METHODS</b>Male wistar rats were given intraperitoneal injection of porcine serum twice a week. At the beginning, the rats in HanDanGanLe-treatment group were feed with HanDanGanLe for precaution. The rats were killed after twelve weeks, then CTGF mRNA and TGFbeta1 mRNA were detected in liver samples with in situ hybridization, and the formation of liver fibrosis was observed with HE stain. The semi-quantitative</p><p><b>RESULTS</b>of the two genes expression were analysed along with the stages of hepatic fibrosis.</p><p><b>RESULTS</b>Typical liver fibrosis developed in the model group rats, and the positive stain of CTGF mRNA and TGFbeta1 mRNA increased, which was distributed in the areas where fibrosis occurred. There was obvious correlation between the expression strength of CTGF mRNA and TGFbeta1 mRNA (r = 0.799, P < 0.05). In the rats receiving HanDanGanLe, CTGF mRNA expression index decreased markedly (12.5+/-2.3 vs 28.8+/-1.4, t = 5.208, P < 0.01), so did TGFbeta1 mRNA expression index (25.4+/-3.2 vs 37.3+/-5.4, t = 5.655, P < 0.01). There was also significant correlation between the scores of CTGF mRNA expression and the stages of hepatic fibrosis (rs = 0.822, 0.808 in the model group and HanDanGanLe-treatment group, P < 0.05).</p><p><b>CONCLUSIONS</b>The expression of CTGF mRNA and TGFbeta1 mRNA is correlated closely with hepatic fibrosis degree. HanDanGanLe can effectively prevent the expression of CTGF and TGFbeta1. One of the mechanisms of the intervention may be its blocking the intracellular signalling pathways involved in liver fibrogenesis.</p>