RESUMO
Serum liver enzyme levels are often used effectively for the evaluation of nonalcoholic fatty liver disease (NAFLD). We aimed to investigate the associations between serum liver enzyme levels and risks for NAFLD in over 8000 cases in a large-scale analysis. A cross-sectional survey with multiple stages and random samplings was performed from May 2007 to May 2009 on 8102 workers at Tongji University. A questionnaire was given, assessments of physical measurements, plasma glucose, lipid profiles, and liver enzymes were made, and real-time liver ultrasounds conducted. The prevalence of NAFLD in Tongji University was 22.2%. It was higher in males than in females (P = 0.0023). The body mass index, waist-to-hip ratio, serum total triglycerides, serum total cholesterol, alanine aminotransferase (ALT), aspartate aminotransferase (AST), and gamma-glutamyl transferase (GGT) values were all higher in the NAFLD group than in the control group. For moderate and severe NAFLD patients, the ALT, AST and GGT values were significantly increased, high density lipoprotein cholesterol was decreased, and drinking much, heavy entertainment and less exercise were more prevalent (P < 0.001). There were strong correlations between serum liver enzyme levels and NAFLD (P < 0.001), with GGT being a more sensitive marker for NAFLD than ALT or AST. ALT and GGT were independent predictors for NAFLD, and GGT was a better predictor than ALT for NAFLD.
Assuntos
Fígado/enzimologia , Fígado/patologia , Hepatopatia Gordurosa não Alcoólica/enzimologia , Hepatopatia Gordurosa não Alcoólica/etiologia , Adulto , Estudos de Casos e Controles , Feminino , Humanos , Testes de Função Hepática , Masculino , Hepatopatia Gordurosa não Alcoólica/diagnóstico , Hepatopatia Gordurosa não Alcoólica/epidemiologia , Prevalência , Fatores de Risco , Índice de Gravidade de DoençaRESUMO
This study was designed to show whether rat liver epithelial cells could undergo epithelial-mesenchymal transition (EMT), thereby directly contributing to liver fibrosis. The role of the ratio of transforming growth factor-ß1 (TGF-ß1)/bone morphogenetic protein-7 (BMP-7) was evaluated in the progression of EMT or mesenchymal-epithelial transition. Primary rat liver epithelial cells were stimulated with different ratios of TGF-ß1/BMP-7 and examined for evidence of transition to a mesenchymal or epithelial phenotype. Liver sections were labeled to detect antigens associated with liver epithelial cells [E-cadherin (E-cad)], EMT [fibroblast-specific protein-1 (FSP-1), vimentin], myofibroblasts [α-smooth muscle actin (α-SMA)], and intracellular signal-transduction mediated by forming liver fibrosis undergo EMT, resulting in the formation of invasive fibroblasts; this process may be driven or impeded by a response to local TGF-ß1 or BMP-7. BMP-7 downregulated α-SMA and phosphorylated Smad2/3. Stimulation of cultured cells with TGF-ß1 induced the expression of pSmad2/3, FSP-1, and α-SMA. Stimulation of cultured cells with BMP-7 induced the expression of E-cad. We demonstrated that the cells upregulated E-cad release compared with untreated cells, but TGF-ß1 was different. We found that the equilibrium of the ratio of TGF-ß1/BMP-7 was 1/10. In summary, the mechanism for this process was not determined. Demonstration of the contribution of what the ratio of TGF-ß1/BMP-7 induced to EMT to the chronic liver diseases would provide a new basis for understanding pathogenesis and potential treatment.
Assuntos
Proteína Morfogenética Óssea 7/metabolismo , Transição Epitelial-Mesenquimal , Fígado/patologia , Fator de Crescimento Transformador beta1/metabolismo , Actinas/metabolismo , Animais , Caderinas/genética , Caderinas/metabolismo , Proteínas de Ligação ao Cálcio/metabolismo , Células Cultivadas , Modelos Animais de Doenças , Células Epiteliais/metabolismo , Fibrose , Regulação da Expressão Gênica , Fígado/metabolismo , RatosRESUMO
To investigate the variance of exogenous gene expression driven by different promoters by in vivo electroporation, 3 plasmid vectors carrying different promoters were selected, and their driving strength was compared in developing chicken embryos. The 3 promoters included: 1) the CAG promoter (containing the cytomegalovirus (CMV) immediate early enhancer and the chicken ß-actin promoter), 2) the CMV promoter (the human CMV immediate early region enhancer), and 3) the SV40 promoter (Simian virus 40). The intensity of GFP expression driven by the 3 promoters was detected by fluorescence microscopy. The results clearly showed that the expression intensity of the reporter gene differed significantly among the 3 promoters. Chicken ß-actin promoter induced the highest intensity of GFP expression, while SV40 promoter induced the lowest intensity. Our results indicate that plasmids with appropriate promoters should be carefully selected to obtain strong exogenous gene expression by in vivo electroporation.
Assuntos
Galinhas/metabolismo , DNA Viral/análise , Proteínas de Fluorescência Verde/metabolismo , Plasmídeos/genética , Animais , Embrião de Galinha , Galinhas/crescimento & desenvolvimento , Eletroporação , Expressão Gênica , Vetores Genéticos/genética , Proteínas de Fluorescência Verde/genética , Microscopia de Fluorescência , Regiões Promotoras Genéticas , Medula Espinal/ultraestruturaRESUMO
Protocadherins constitute a large family belonging to the cadherin superfamily; they function in various tissues of a wide variety of multicellular organisms. However, their functions and expression modes are still unknown in many of these species and tissues. We developed a fast and low-cost method to produce polyclonal antibody against chicken protocadherin 1 (Pcdh1) that could be used in assays for immunological assessment of protein expression levels of chicken Pcdh1. Primers were designed with DNAStar, using the nuclear sequence of pcdh1 as a template; the pcdh101 fragment was amplified, identified by sequencing and cloned into expression vectors pGEX-2TK and pET-32a, separately, resulting in 2 recombinant plasmids, pGEX-2TK-pcdh101 and pET-32a-pcdh101. These were confirmed by double-enzyme digestion and sequencing. The recombinant expression vectors were transformed and expressed in Escherichia coli BL21. The recombinant oligopeptides glutathione-S-transferase (GST)-Pcdh101 and (His)6-Pcdh101 fused with the carrier protein GST and (His)6 separately, and were purified. Rats were immunized by injecting the emulsified GST-Pcdh101 antigen subcutaneously into their hind footpads, followed by a booster injection after 2 weeks. One week after the booster, the sera were collected and examined for antibody titer by indirect ELISA. The optimal dilution of this antiserum was 1:300. The specificity of the antiserum was confirmed by Western blotting. This antiserum had good specificity and could be used to detect chicken Pcdh1 in Western blot analysis. This method allows production of specific rat polyclonal antisera for Western blots in less than 1 month at a relatively low cost.