Objective To investigate the changes of human epidermal stem cells after transfected with human telomerase reverse transcriptase(hTERT)gene.Methods The plasmid pIRES2-EGFP and plasmid pIRES2-EGFP-hTERT encoding hTERT were transfected into in vitro cultured human fetal epidermal stem cells by liposome-mediated transfection.Then,the positive cells were selected with G418.The mRNA and protein expressions of hTERT were detected by reserve transcriptase-polymerase chain reaction(RT-PCR)and Western blot.The telomerase activity and the proliferation and cycle of human epidermal stem cells were detected by telomeric repeat amplification protocol(TRAP)-ELISA and flow cytometry respectively.Results RT-PCR and Western blot techniques detected weak mRNA and protein expressions of hTERT gene in untransfected and vacant vector transfected cells but high level of mRNA and protein expressions of hTERT gene in pIRES2-EGFP-hTERT transfected cells.Compared with untransfected and vacant vector transfected cells,the pIRES2-EGFP-hTERT transfected cells had higher telomerase activity,with lower proportion of cells at G_0/G_1 phase,higher proportion of cells at S and G_2/M phases and enhanced proliferation ability.Conclusion Transfection with hTERT gene can markedly enhance mRNA and protein expressions,telomerase activity and proliferation ability of hTERT gene of human epidermal stem cells euhured in vitro.

Objective To study the significance of expressions of smad_4mRNA,TGF-?_1, and TGF-?R_1 in pancreatic carcinoma(PC) . Methods Smad_4mRNA was detected by in situ hybridization. TGF-?_1 and TGF-?R_1 were detected by immunohistochemical method. Results The positive rates of smad_4mRNA,TGF-?_1 and TGF-?R_1 were singnificantly lower in 53 slices of pancreatic carcinoma than those in 25 slices of paracancerous tissue (all P

Objective Investigate the Protective effect of paeoniflorin in rats with acute liver injury. Methods Male SD rats were randomly divided into normal group, model group, paeoniflorin in small, middle and high dose group (20 mg/kg, 40 mg/kg, 80 mg/kg), paeony glycoside group (50 mg/kg). Except normal group, the rest of groups were irradiated fractionally by VARIN 21-EX linear accelerator at right liver, The paeoniflorin group and paeony glycoside group were lavaged everyday after irradiation for corresponding drugs and doses. Normal group and model group give equal volume normal saline everyday. All rats were killed on 2nd and 4th weekend. Measure rats serum AST, ALT, hepatic tissue GSH, SOD, and HE staining score. Results The rats in model group liver tissue HE staining scores increased to(2.25±0.53)on 2nd weekend, The serum levels of AST, ALT increased to(112.83±19.20)U/L, (80±21.97)U/L, it significantly increased Compared with the normal group(63.06±7.15)U/L, (42.30±4.45)U/L, P<0.01. The liver GSH contents of paeoniflorin in each dosage groups rats were(60.89±8.43)U/mg, (67.84±9.05)U/mg, (71.92±8.11)U/mg on 2 nd weekend, Compared with the model group(37.32±11.25)U/mg, (90.54±12.12)U/mg, P<0.05或<0.01. Conclusions The irradiated rats go into acute liver injury on 2nd weekend, paeoniflorin has protective effect on acute liver injury in rats.

Objective To assess the effect of intravenous infusion of octreotide in prevention of pancreatic fistula after pancreaticoduodenectomy. Methods The clinical data of 74 cases of pancreaticoduodenectomy performed from January 1996 to July 2003 were retrospectively reviewed. These included 36 cases in control group in which octreotide was not adminstered,and 38 cases in octreotide group in which octreotide was administered by intravenous infusion of 0.5?g/( kg?h) for 12 hours per day from the operative day to postoperative day 7. The study parameters included clinical manifestation,drainage from peritioned cavity and the amount of drainage of pancreatic fluid. Results The drainage of pancreatic fluid at postoperative day 1,3,5 the in octreotide group was significantly less than those in the control group,the average hospital stay and the incidence of pancreatic fistula were significantly lower than those in the control group,and the drainage of pancreatic fluid was significantly increased after the withdrawal of octreotide in the octreotide group. Conclusions Intravenous infusion of octreotide can significantly lower the incidence of pancreatic fistula after pancreaticoduodenectomy.

Objective To evaluate the effect of surgical therapy for hepatolithiasis by hepatectomy and choledochofiberscopy. Method We retrospectively analyzed hepatolithiasis patients in our hospital during the past 5 years, comparing the therapeutic effect of different surgical modality. Data were analyzed by chi-square test. Result A total of 469 patients underwent surgical treatment, 412(87.85%) cases were followed up for an average of 3 years and 6 months. The residual calculus rate was 5. 14% and 14. 81% (x2 =9.32,P

Objective To study the effect of expression of myoglobin which mediated by adenovirus,on ATP value of liver and the protective effect on liver ischemia reperfusion injury.Methods Adenovirus carrying CMV promoter sequences linked to the human myoglobin gene(AdCMVMyo) were transfected into rats liver. Then myoglobin, hepatic ATP levels and liver function were evaluated. Results Myoglobin expression was verified in rat livers after AdCMVMyo transfection. The ATP levels in rat livers 72 hours after AdCMVMyo transfection were significantly higher than that in control group(P

Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated proteins (Cas) system is a hotspot of gene editing and gene expression research, in which CRISPR/Cas13 system provides a new direction for RNA interference and editing. In this study, we designed and synthesized the corresponding gRNAs of CRISPR/Cas13a and CRISPR/Cas13b systems in non-homologous end joining (NHEJ) pathway, such as Ku70 and Lig4, and then detected the expression of ku70 and lig4 in HEK293T cells. The CRISPR/Cas13a system could efficiently knockdown the mRNA expression of ku70 and lig4 more than 50%, and CRISPR/Cas13b system also suppressed ku70 and lig4 about 92% and 76%, respectively. Also, CRISPR/Cas13a, b systems could down-regulate Ku70 and Lig4 proteins level to 68% and 53%, respectively. The study demonstrates that the CRISPR/Cas13 system could effectively knockdown the expression of RNA and protein in HEK293T cells, providing a new strategy for gene function and regulation research.
CRISPR-Cas Systems ; DNA Ligase ATP ; genetics ; Gene Expression Regulation ; genetics ; Gene Knockdown Techniques ; HEK293 Cells ; Humans ; Ku Autoantigen ; genetics