A hospital-based case-control study on influencing factors of nonalcoholic fatty liver disease / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To investigate the influencing factors of nonalcoholic fatty liver disease (NAFLD).</p><p><b>METHODS</b>A hospital-based case-control study was conducted in patients with NAFLD and controls without NAFLD in a hospital from January to August in 2007. All data were analyzed by SPSS 13.0 software.</p><p><b>RESULTS</b>One-way analysis of variance found that the two groups were significantly different in cigarette smoking, alcohol and tea comsumption, movement index, speed of food intake, frequency of social engagement, kinds of edible oil, marine products, family history of NAFLD, hypertension, higher blood sugar, abnormality of blood fat, higher level of ALT, higher level of AST, hyperuricemia, obesity, decrease of high density lipoprotein (HDL), and increase of low density lipoprotein. By non-conditional logistic stepwise regression analysis, 12 of 18 factors were used to construct a model, ten of which were the risk factors and two were protective factors of NAFLD. Risk factors included obesity (OR=6.35), hypertension(OR=3.82), dyslipidemia (OR=2.95), decrease of HDL (OR=2.85), hyperglycemia (OR=2.82), increase of ALT (OR=2.80), hyperuricemia (OR=2.35), HBsAg positive (OR=1.99), family history of fatty liver (OR=1.79) and frequently intake of marine products (OR=1.58), and protective factors included tea drinking (OR=0.72) and exercise (OR=0.90).</p><p><b>CONCLUSIONS</b>There are many influencing factors of NAFLD, and life styles are the key factors. Genetic background may also play some roles in NAFLD.</p>

Stable replication and antigen expression of hepatitis B virus in HepG2 cells / 中华传染病杂志

Objective To establish the HepG2 cell lines which can stably express and replicate hepatitis 13 virus (HBV).Methods One point two X unit length of HBV genome was cloned intn SalⅠsite of the eukaryotic expression vector pREP10 to construct the recombinant plasmid pREP-HBV. Human hepatoblastoma cell HepG2 was transfected with pREP-HBV by Lipofectamine 2000 and seh,cted by bygromycin at the concentration of 250?g/mL.HBsAg and HBeAg were monitored by enzyme linked immunosorbent assay (ELISA)kits.H13V particles presemed in supernatant were ex- amincd by electronic microscopy.DNA isolated from intracellular HBV core particles was analyzed by Southbern blot using HBV-specific probe.Results The recombinant vector pREP HBV containing 1.2?unit length of HBV DNA was constructed successfully.After transfection of pREP-HBV to HepG2 cells and consistently cultured in hygromycin selective medium.5 drug-resistant cell lines, RHBV1-RHBV5.were established,and all of them could stably express HBsAgand HBeAg.South ern blot analysis revealed that HBV could replicate in all cell lines,as confirmed by the presence of replicateintermediatc DNA in intracellular HBV core particles.Clustered 42 nm Dane particles as well as 22-26 nm spherical H13sAg particles in condensed cuhure supernatant were visualized by elec tronic microsopic analysis.Conclusion HepG2 ceil lines in which HBV can replicate and express specific antigens are successfully established.Up to now,the cells have been passaged every three days for 50 times.

Influence of enterococci on human sperm membrane in vitro / 亚洲男科学杂志(英文版)

<p><b>AIM</b>To study the influence of enterococci on human sperm membrane in vitro.</p><p><b>METHODS</b>Ejaculated human sperm were artificially infected with beta-hemolytic or non-beta-hemolytic enterococci at the bacteria: sperm ratio of 50:1 at 37 degrees . Sperm membrane integrity was examined after incubation for 1, 3 and 5 h by hypoosmotic swelling (HOS) test and electron microscopy.</p><p><b>RESULTS</b>Sperm infected with beta-hemolytic enterococci had lower HOS scores compared with non-beta-hemolytic strains or uninfected control (P < 0.01). The HOS test scores of sperm infected with beta-hemolytic enterococci increased in the presence of phosphatidylcholine, an inhibitor of hemolysin. Non-beta-hemolytic strains showed no significant difference in swelling rate, compared with the control group (P > 0.05). It was shown by electron microscopy that beta-hemolytic enterococci caused significant rupture of human sperm membrane.</p><p><b>CONCLUSION</b>Beta-hemolytic enterococci caused human sperm membrane injury, and might be mediated by the hemolysin of enterococci.</p>

Expression of Recombinant Snake Venom Cystatin in Yeast Pichia pastoris and Its Effects on B16F1 Melanoma Invasion in vitro / 中国生物工程杂志

To investigate the biological role of snake venom cystatin(sv-cystatin) in tumor progression, the cDNA of sv-cystatin amplified by PCR from pUC18-cystatin plasmid was cloned into methanol-inducible expression vector pPICZ?A. The linearized recombinant plasmid pPICZ?A-cystatin was transfered into Pichia pastoris, strain GS115 by electrophoration. Transfermants with phenotype Mut+ selected were identified by PCR analysis and induced in 1.0% methanol. The reombinant sv-cystatin protein was examined by SDS-PAGE, Western blot analysis. The molecular mass of expression product was about 14 kDa and approximately 16 mg/L of recombinant sv-cystatin was produced from one of GS115-cystatin transformants. The chromatography purified protein could reduce the activity of papain. The ability of B16F1 cells treated with recombinant sv-cystatin to invade the reconstituted basement membrane decreased significantly (P

Correlation between the expression of angiopoietins and their receptor and angiogenesis in gastric cancers / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To explore the effects of angiopoietins (Ang-1 and Ang-2) and Tie-2 expression on microvessel density (MVD) in gastric cancers.</p><p><b>METHODS</b>By using semiquantitative RT-PCR, immunohistochemistry and image analysis system, the expression of Ang-1, Ang-2, Tie-2 mRNA and their proteins were detected in 68 primary gastric cancers and their adjacent normal tissues. Microvessel density (MVD) was figured out based on CD34 immunohistochemical staining.</p><p><b>RESULTS</b>The expression of all Ang-1, Ang -2, Tie-2 mRNA and their proteins was detected in gastric cancers and their paired adjacent gastric mucosa tissues. A negative correlation between Ang-1 protein, Tie-2 mRNA and MVD in gastric cancers was observed (r = -0.440, r = -0.267; P < 0.05), while the relation between Ang-2 mRNA and its protein, Ang-2/Ang-1 protein ratio with MVD were positive (r = 0.319, r = 0.729, r = 0.739; P < 0.05). It was found that MVD in groups with Ang-2 mRNA T/N ratio over 1.2 (the ratio of Ang-2 mRNA in gastric cancers and its adjacent normal mucosa) was higher than that in those with a ratio under 1.2, revealed by analysing the effects of Ang-1 and Ang-2 mRNA T/N ratio on MVD in gastric cancers.</p><p><b>CONCLUSION</b>Ang-1 activates Tie-2 receptor, whereas Ang-2 antagonizes Ang-1 in the angiogenesis, and the Ang-2/Ang-1 ratio determines angiogenesis and tumor growth in gastric cancers. When the expression of Ang-2 is high and Ang-1 is low, the angiogenesis in gastric cancers is promoted, otherwise oppositely. The role of Ang-2 is dominant in the effect of Angs and their receptor on angiogenesis in gastric cancers.</p>

Dual effect of angiopoietin-2 on angiogenesis in gastric cancer / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To explore the effect of Ang-2 on angiogenesis in gastric cancer.</p><p><b>METHODS</b>The expression of Ang-2 mRNA was analyzed by RT-PCR and the expression of VEGF and CD34 was detected by immunohistochemistry in 36 cases of gastric cancer tissues and their paired adjacent gastric mucosa.</p><p><b>RESULTS</b>The expression of Ang-2 mRNA was found both in gastric cancer and their paired adjacent gastric mucosa; the correlationship between the general expression levels of Ang-2 mRNA and microvessel density (MVD) in gastric cancer tissues was not found. However, in 27 cases whose Ang-2 mRNA expression levels in cancer tissues were lower than those in adjacent gastric mucosa. A significant positive correlation between the expression level of Ang-2 mRNA and MVD in the tumor tissues was found (r = 0.411, P < 0.05). In these 27 cases, the MVD in the gastric cancer tissues with positive VEGF expression (45.45 +/- 10.30) was higher than that with negative VEGF expression (30.15 +/- 8.69, P < 0.05), whereas in the other 9 cases whose expression levels of Ang-2 mRNA in cancer tissues were higher than those in adjacent gastric mucosa, a significant negative correlation between expression level of Ang-2 mRNA and MVD in the tumor tissues (r = -0.758, P < 0.05), but without correlation between the MVD and VEGF.</p><p><b>CONCLUSION</b>Under the conditions that the expression of Ang-2 mRNA in cancer tissues was lower than that in adjacent gastric mucosa, VEGF could promote the sprouting of new vessels along with Ang-2 upregulation. But under the conditions that the expression of Ang-2 mRNA in cancer tissues was higher than that in adjacent gastric mucosa, Ang-2 inhibited angiogenesis. Angiopoietin-2 may play a dual effect on angiogenesis in gastric cancer.</p>

Effects of nitric oxide on mitochondrial permeability transition and cytochrome C of human hepatocellular carcinoma cell lines / 中国医学科学院学报

<p><b>OBJECTIVE</b>To investigate the effects of nitric oxide on mitochondrial permeability and cytochrome C (cyt C) of human hepatocellular carcinoma cell lines.</p><p><b>METHODS</b>NO-mediated apoptosis in human hepatocellular carcinoma cell lines SMMC-7721 and HepG2 was investigated by flow cytometry. The growth and proliferation of human hepatocellular carcinoma cell lines SMMC-7721 and HepG2 were evaluted by MTT assay. Mitochondrial transmembrane potential was analyzed by flow cytometry with double staining of Rh123 and PI, and cytoplasmid cyt C was measured by Western blot. The cells were preincubate with cyclosporin A or GSH synthesis blocker BSO to explore their effect on the results of the above experiments.</p><p><b>RESULTS</b>NO donor sodium nitroprusside (SNP) induced apoptosis in human hepatocellular carcinoma cell lines SMMC-7721 and HepG2 and resulted in the decrease of the mitochondrial transmembrane potential and the increase of the amount of cytoplasmid cyt C in time-dependent manner. Cyclosporin A (CsA) specific inhibitor of the mitochondrial permeability transition pore could partially prevent the decrease of delta psi m and the release of cyt C. In contrast, GSH synthesis blocker BSO promoted the decrease of delta psi m and the release of cyt C.</p><p><b>CONCLUSIONS</b>NO may induce apoptosis in human hepatocellular carcinoma cell lines SMMC-7721 and HepG2 by decreasing delta psi m, opening the mitochondrial permeability transition pore, and releasing the cyt C.</p>

Structural analysis of 22 full-length hepatitis B virus genomes isolated from patients with hepatocellular carcinoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To study the characteristics of full-length Hepatitis B Virus (HBV) genomes isolated from patients with hepatocellular carcinoma (HCC).</p><p><b>METHODS</b>The full-length HBV genomes from the serum of HBsAg positive HCC patients were amplified by PCR, and then sequenced and analyzed its structure.</p><p><b>RESULTS</b>Twenty-two full-length HBV DNAs were obtained from different patients of HCC. Phylogenetic analysis revealed that all HBV strains could be categorized into genotype B or C and serotype adr or adw2. Structural analysis showed that HBV obtained shared meaningful consensus mutations in B/T cell epitopes of surface and core antigens, transactivating domain of X protein and enhancer II/core promoter regions as compared to standard strains.</p><p><b>CONCLUSION</b>Genotype and gene mutation of HBV may be closely correlated with the carcinogenesis of HBV-related hepatocellular carcinoma.</p>

Expression and significance of nitric oxide synthase in human hepatocellular carcinoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the expression and significance of nitric oxide synthase (cNOS) mRNA in primary hepatocellular carcinoma (HCC), cirrhotic liver and normal liver tissue.</p><p><b>METHODS</b>cNOS mRNA expression in 80 HCC, 40 cirrhotic liver and 20 normal liver tissue were observed by in situ hybridization. CD34 immunostain was used to measure the microvascular density (MVD) and Ki-67 immunostain to proliferative index.</p><p><b>RESULTS</b>Expression of cNOS mRNA was observed in the liver cancer cells, endothelial cells in the non-cancerous liver tissues and mononuclear and/or phagocytes. Expression of cNOS mRNA in tumor cells of HCC was higher than that in the liver cells of cirrhotic liver (P < 0.01) which was higher than the normal liver tissue. Expression in the endothelial cells was higher in HCC and cirrhotic liver than those in the normal liver tissue (P < 0.01). HCC with positive cNOS mRNA expression in the endothelial cells showed higher extent of neovascularization and degree of proliferative index. The more MVD, the higher proliferative index, which increased in metastatic tumors.</p><p><b>CONCLUSION</b>cNOS mRNA expression was involved in oncogenesis, angiogenesis and progression of hepatocellular carcinoma.</p>