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Objective To investigate the efficiency of Trichostatin A (TSA) in inducing cell apoptosis and altering the Notch pathway genes expression in PANC-1 cells line.Methods The survival rate and apoptosis of PANC-1 cells were measured by MTT assay and Hoechst 33258 staining,respectively.mRNA expression levels of the genes,numb,gcn512,dll3,hes6,eaf2,cytohesins,in PANC-1 cells were assessed by real-time quantitive PCR.Western blot was used to measure the expression of bcl-2,bax,actived caspase-3 and NICD protein which was the biologically active form of Notch-1.Results After culturing with 0.1,0.2,and 0.4 μmol/L TSA for 24 hours,the cellular survival rate of PANC-1 cells significantly decreased to 72％,58％ and 39％,respectively.The survival rate of PANC-1 was negatively correlated to time length of culture with TSA.Increased apoptosis of PANC-1 cells after 12,24 and 36 h culture with TSA was detected by Hoechst 33258 staining.Western blotting showed that the expression of bax,actived caspase-3 and NICD protein increased while the bcl-2 protein decreased after culture with TSA.In real time quantitive PCR assessment,the mRNA expression of numb and hes6 in PANC-1 cells were upregulated by TSA (P ＜ 0.05),while the mRNA expression of gcn512 and dll3 were down-regulated by TSA (P ＜ 0.05).While mRNA expressions of eaf2 and cytohesin1,2,3,4 were not affected by TSA.Conclusions TSA induces apoptosis of pancreatic cancer cell line PANC-1.The Notch signal pathway may be involved in inducing cellular apoptosis of PANC-1 when cultured with TSA.
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Objective To observe the transfection efficiency and anti-fibrotic effect of miR-29b transfected by anti-TGF-β Ⅱ R ScFv/Ck/tP fusion protein (new vector) in hepatic stellate cell (HSC),and to provide a new vector in gene therapy for liver fibrosis.Methods The liposome vector,new vector,and lentiviral vector were used as transfection reagents to transfect miR-29b into HSC.Transfection efficiency was observed under fluorescence microscope and flow cytometry.Collagen α1 (Ⅰ) mRNA and protein expression in different groups were analyzed by real-time RT-PCR and Western Blot,respectively.Results Compared to the control,transfection efficiencies in lentiviral vector,new vector,and liposome vector groups were about 70％,58％,and 29％,respectively.Collagen α1 (Ⅰ) mRNA expression in lentiviral vector,new vector,and liposome vector groups was decreased by about 70％,50％,and 38％,respectively ((t =6.316,P ＜0.01 ; t =4.082,P ＜0.01 ; t =3.014,P ＜0.05).Collagen α1(Ⅰ) protein expression in lentiviral vector,new vector,and liposome vector groups was decreased by about 59％,41％,and 27％,respectively (t =4.209,P ＜0.01; t =4.033,P ＜0.01; t =2.842,P ＜0.05).Conclusions The new vector constructed by us has a high transfection efficiency.MiR-29b transfected by the new vector has a good anti-liver fibrosis effect.
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Objective To explore the factors related to the delayed graft function (DGF). Methods Clinical data of 150 recipients were collected and performed by Cox proportional hazards regression analysis . In addition, the glutathione S-transferase (GST) gene polymorphism of 172 donors and 157 healthy persons was analyzed by multiple PCR and SSP-PCR. Results DGF was observed in 24 patients among 150 recipients. Pretranplantation dialysis mode, PR A levels and recipient gender were uncorrelated with the incidence of DGF(P>0. 05). Urinary volume of the second 24 hours after transplantation was an independent predictor of DGF(RR=1. 002, P = 0. 001). The frequency of donor's null GSTM1 in DGF group was significantly higher than that in non-DGF group(P<0. 05). Conclusions Urinary volume of the second 24 hours after transplantation could be a predictor for DGF. The null GSTM1 in donor might be one of the factors related to the EGF.
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Objective To investigate the association of genetic polymorphisms in glutathione S-transferases T1 (GSTrl), M1 (GSTM1) and P1 (GSTP1) with aristolochic acid nephropathy (AAN) of Chinese people in Wenzhou of China. Methods Fifty-nine patientswith AAN (AAN group) including 29 male and 30 female as well as 157 healthy ethnically matched controls (control group) including 93 male and 64 female were enrolled in this study. The genotypes of GSTT1, GSTMI and GSTP1 were determined by multiple PCR and confronting two-pair primers PCR (CTPP-PCR). Results The genotype frequencies of GSTP1 were in Hardy-Weinberg equilibrium. Compared with the healthy controls, the frequency of GSTT1 null genotype was significantly higher in the patients with AAN (66.1% vs 48.4%,P＜0.05). Risk of A.AN for individuals with GSTT1 null genotype was 1.747 fold of those without GSTIl null genotype (95% CI=0.818-3.731). The frequency of GSTM1 null genotype, GSTP1 variant genotypes and GSTP1 G allele in the patients and in the controls were 40.7%, 28.8%, 16.1% and 47.8%, 31.8%, 17.5%, respectively, which were not significantly different. No significant differences were found in prevalence of GSTM1 and GSTP1 gene distribution between patients and controls. Conclusion GSTrl gene polymorphism appears to be associated with susceptibility to AAN in Southern China.
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0.05).The optimal combination was PCR-CTPP for MDR1 C3435T and PCR-SSP for G2677T/A.Conclusions PCR-CTPP and PCR-SSP are simple,accurate,rapid and economical methods for detection of SNP of MDR1 C3435T and G2677T/A,and can be applied in clinical research.