Inhibitory effect of siRNA-YAP1 on transforming growth factor β 2-induced epithelial-mesenchymal transition in human lens epithelial cells / 中华实验眼科杂志

Objective:To investigate the inhibitory effect of small interfering RNA-Yes-associated protein 1 (siRNA-YAP1) on epithelial-mesenchymal transition (EMT) in human lens epithelial cells (LECs) induced by transforming growth factor-β 2 (TGF-β 2). Methods:Human LECs line (HLEB-3) was cultured and divided into normal control group and TGF-β 2 induced group.The cells in the normal control group were treated with serum-free low-glucose medium for 24 hours, and the cells in the TGF-β 2 induced group were treated with additional 10 ng/ml TGF-β 2 for 24 hours.The cultured HLEB-3 cells were divided into siRNA empty vector group, siRNA-YAP1 transfection group, siRNA empty vector+ TGF-β 2 group and siRNA-YAP1+ TGF-β 2 group, and the cells were transfected with plasmid including siRNA empty vector or siRNA-YAP1 sequence according to grouping.The relative expression levels of YAP1 mRNA and protein in various groups were detected and compared by quantitative real-time polymerase chain reaction (PCR), immunofluorescence and Western blot assay, respectively.The relative expression levels of EMT marker proteins (E-cadherin and Vimentin proteins) in various groups were detected by immunofluorescence and Western blot assay. Results:Compared with the normal control group, the expression level of E-cadherin protein was decreased (1.180±0.118 vs.0.830±0.104) and the Vimentin protein was increased (0.797±0.110 vs.1.240±0.110) in the TGF-β 2 induced group, with significant differences between the two groups ( t=3.857, P=0.018; t=-4.933, P=0.008).The relative expression levels of YAP1 mRNA and protein in the TGF-β 2 induced group were significantly increased in comparison with the normal control group (2.200±0.193 vs.1.136±0.123; 1.203±0.121 vs.0.967±0.025), with significant differences between the two groups ( t=-9.288, P<0.01; t=-3.329, P=0.029).Compared with the siRNA empty vector group, the expression levels of YAP1 mRNA and protein in the siRNA-YAP1 transfection group were significantly reduced (both at P<0.01).Compared with the siRNA empty vector+ TGF-β 2 group, the relative expression level of E-cadherin protein was significantly enhanced and the expression level of Vimentin protein was significantly reduced in the siRNA-YAP1+ TGF-β 2 group (both at P<0.01). Conclusions:YAP1 participates in the TGF-β 2 induced EMT in human LECs, and siRNA-YAP1 can suppress the EMT process.

Clinical application of interleukin-27 in the patients with acute coronary syndrome / 临床检验杂志

Objective To investigate the application value of interleukin-27 (IL-27) in the patients with acute coronary syndrome (ACS).Methods A total of 208 ACS patients were enrolled in the study,including 76 acute myocardial infarction (AMI) patients with ST elevation (STEMI),58 AMI patients with non-ST elevation (NSTEMI) and 74 unstable angina pectoris (UAP) patients.These patients were divided into single-vessel lesions,double-vessel lesions and three-vessel lesions groups,and 62 patients with chest pain syndrome (CPS) were selected as a control group.The plasma IL-27 levels of all patients were determined by enzyme linked immunosorbent assay (ELISA) and analyzed.Results The levels of plasma IL-27 (median[P25,P75]) in STEMI (308.64 [245.17,359.26] pg/mL),NSTEMI (256.88 [181.52,332.51] pg/mL) and UAP (218.12 [165.33,312.46] pg/mL) patients were significantly higher than that in CPS patients (100.66[68.98,228.86] pg/mL,P ＜ 0.01).The levels of plasma IL-27 in STEMI patients were significantly higher than that in NSTEMI and UAP patients (P ＜ 0.05).The positive rate of IL-27 in ACS patients with negative TnI was 54.24％ (32/59).The sensitivity and specificity of IL-27 in predicting ACS from chest pain patients were 80.29％and 58.06％,respectively.The levels of plasma IL-27 in the patients with three-vessel lesions were significantly higher than that with single-vessel lesions (P ＜ 0.05).Conclusion Plasma IL-27 levels in ACS patients increase obviously,which may be involved in the pathogenesis of ACS.IL-27 may be helpful for the diagnosis of ACS patients with negative TnI and the prediction of ACS state.

Carboxymethytl pachymaram induces the methylation of SOCS-1 gene and the maturation of human monocyte-derived dendritic cells / 中华微生物学和免疫学杂志

Objective To investigate the effects of carboxymethytl pachymaram ( CMP ) on the methylation of SOCS-1 (suppressor of cytokine signaling-1) gene and the in vitro maturation of human mono-cyte-derived dendritic cells (DCs).Methods Human DCs were induced from the peripheral blood mono-cytes in vitro with the treatment of recombined human GM-CSF and interleukin-4 ( IL-4 ) and cultured with different concentrations of CMP (10, 50, and 100 mg/L).The methylation and expression of SOCS-1 gene were analyzed by methylation-specific polymerase chain reaction (MSP) and real-time PCR, respectively. The phenotypic markers of DCs were detected by flow cytometry .Mixed lymphocyte reaction ( MLR) and ELISA were performed to measure the lymphocyte proliferation induced by DCs and IL-12 secretion by DCs . Results CMP promoted the methylation of SOCS-1 gene, but inhibited the expression of SOCS-1 gene in dendritic cells at the concentrations of 50 mg/L and 100 mg/L.The expression of phenotypic markers (CD80, CD83, CD86 and HLA-DR), IL-12 secretion and lymphocyte proliferation induced by DCs were significantly enhanced in a dose dependent manner with the treatment of CMP .Compared with control group , the levels of methylated SOCS-1 gene and IL-12 and the lymphocyte proliferation index were increased upon the stimulation with 50 mg/L and 100 mg/L of CMP (P<0.01), but the expression of SOCS-1 gene was de-creased.The expression of CD80, CD83 and HLA-DR on DCs in the presence of 100 mg/L of CMP were higher than those of control group (P<0.05).Conclusion CMP could induce the methylation of SOCS-1 gene and the maturation of DCs derived from peripheral blood monocytes .

Application of comprehensive teaching mode in cataract clinical teaching / 中华医学教育探索杂志

Objective To explore teaching effect of comprehensive teaching mode of PBL combined with CBL in ophthalmologic cataract clinical teaching.Methods 2008 grade clinical majors (n =80) in class 1 as experimental group were taught by comprehensive teaching mode of PBL combined with CBL while those (n =83) in class 2 as control group were taught by traditional LBL teaching mode.After the courses,the teaching effects of two methods were compared.SPSS 11.5 software was used for statistical analysis,x2 test for satisfaction survey and t-test for theoretical examination scores.The test level is α =0.05.Results There were significant differences between experimental group and control group in improving comprehensive quality and developing clinical thinking.Scores of understanding knowledge,case analysis and total scores of experimental group were higher than those of control group (all P ＜ 0.05).Conclusions Comprehensive teaching mode may improve the teaching effect of cataract clinical teaching,but it need to be explored and improved continually in practice.

Mechanism of DADS in the bystander effect of HSV-tk/GCV suicide gene therapy system in lens epithelial cells / 中南大学学报(医学版)

OBJECTIVE@#To investigate the mechanism and effect of diallyl disulfide (DADS) on the bystander effect of herpes simplex virus kinase/ganciclovir (HSV-tk/GCV) suicide gene therapy system which was mediated by recombinant adeno-associated virus (rAAV) in lens epithelial cells.@*METHODS@#Immunohistochemistry method was used to determine the distribution of connexin 43 (Cx43) protein in cultured rabbit lens epithelial cells and rabbit lens epithelial cells transfected by HSV-tk suicide gene. Cx43 protein was measured and analyzed in cultured rabbit lens epithelial cells and rabbit lens epithelial cells transfected by HSV-tk suicide gene that was induced by various DADS. Cell survival was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method.@*RESULTS@#DADS increased the Cx43 protein expression in cultured rabbit lens epithelial cells and rabbit lens epithelial cells transfected by HSV-tk suicide gene, and especially in 50 μmol/L DADS. After combining with DADS, the bystander effect was significantly improved (P<0.05).@*CONCLUSION@#DADS can elevate the expression of Cx43 protein and enhance the bystander effect of HSV-tk/GCV suicide gene therapy system.

Analysis of circulating activated platelet and its significance in patients with cerebral infarction / 国际检验医学杂志

Objective To explore the activation of circulating platelet in patients with cerebral infarction(CI) and its correlation with CI.Methods The fibrinogen receptor(FIB-R) and P-selectin were used as molecular marker of circulating platelets, which were analyzed by flow cytometry in 80 healthy persons and 127 CI patients in acute and rehabinating period.Results The FIB-R expression on circulating platelet of CI patients in dangerous period was significantly higher than that in steady period and healthy persons(P0.05).Conclusion The activation of circulating platelet has a close relationship with CI. FIB-R may be a sensitive molecular marker of circulating activated platelet. It will help to evaluate the severe extent of CI and give an anti-platelet treatment as soon as possible to improve the prognosis of CI through monitoring the FIB-R of CI patients successively.