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Objective Blocking the expression of Survivin with siRNA, and the effects of suppling the proliferation of PC-2 cell and inducing its apoptosis were investigated. Methods Constructed the siRNA against Survivin plasmid expression vector and transfected it into PC-2 cell with lipofectamineTM 2000, the changes of Survivin expression were detected by semi-quantitive RT-PCR and immunohistochemical method, The effect of suppressing the proliferation of PC-2 cell was detected by the method of MTT; the effect of inducing PC-2 cell apoptosis was detected by flow cytometry. Results The sequence specific siRNA can effectively block the expression of Survivin both at themRNA and protein levels, the expression inhibition ratio was 81.25% at mRNA level and 74. 24% at protein level;blocking the expression of Survivin can suppress the proliferation of PC-2 cell significantly, 24, 48 hours after the cell was reseeded, the proliferation inhibition ratio was 28. 00% and 33. 38% respectively; 24, 48 hours after the transfection, 8.46 % and 7.53 % cells were induced to apoptosis respectively. Conclusion The siRNA against Survivin plasmid expression vector constructed in the study can blocking the expression of Survivin in PC-2 cell effectively and specifically; blocking the expression of Survivin can signiilcantly suppress the proliferation of PC-2 cell and induce a certain degree cells apoptosis; RNAi against Survivin is of a certain value in the gene therapy of pancreatic cancer.
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Objective In order to investigate the therapeutic effects of portal arterialization for portal hypertension,portal arterialization and complete shunt(PACS) was applied in canine model of portal hypertension,which was made by thread embolization within the portal vein.Methods A splenectomy,splenic artery and upper portal vein anastomosis,and a complete portal-caval shunt were performed on portal hypertension dogs.The blood pressure and flow of the portal vein including that towards the liver and towards the vena cava were observed.Results The postoperative hepatic inflow,PVF,increased to 180% of the former while PVP increased to 196%;the caval-inflow PVF increased to 130% of the former while PVP decreased to 45.5%.Significant difference existed(P
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Objective To study the effect on liver hemodynamics of portal arterialization and complete shunt(PACS),splenorenal shut(SRS) and peripheral cardia divided vessel(PCDV).Methods The preparation of canine model was made.Group PCDV accepted a splenectomy and peripheral cardia divided vessel,while the group SRS accepted a spleen-renal vein shunt.Group PACS accepted a splenectomy,splenic artery and upper portal vein anastomosis,and complete portal-caval shunt.The blood pressure and flow of the portal system were observed.The hepatic function was also measured before and 2 weeks after the three kinds of operation.Results In the PCDV group,the postoperative PVF decreased in 17% while PVP decreased in 5%.In the SRS group,the postoperative PVF decreased in 51% while PVP decreased in 51%.In the PACS group,the postoperative hepatic inflow PVF increased to 180% of the former while PVP increased to 196%;the caval-inflow PVF increased to 130% of the former while PVP decreased to 46%.The results of PACS group had a magnificent statistic difference comparing with those two traditional operations(P
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Objective To construct active recombinant cas pa ses-3 gene(r-caspases-3)eukaryotic expression plasmid and observe the apoptos is inducing activity of r-caspase-3 in pancreatic carcinoma cells. Methods pcDNA3.1(+)/r-caspase-3 was constructed and pan creatic carcinoma cells(PC-Ⅱ)were transfected with the pcDNA3.1(+)/r-caspases -3 by liposomes(LipofectAMINE).The expression of r-Caspase-3 mRNA in pancreat ic carcinoma cells was detected by reverse transcription process of the polymera se chain reaction(RT-PCR), and the signs of apoptosis were examined in pancreat ic carcinoma cells by the methods of the DNA electrophoresis and flow cytometry analysis(FACS).Results The sequence inserted in pBlueSKM/r-Caspase-3 p lasmid was coincident with that of the r-caspases-3. The evaluation result of pcDNA3.1(+)/r-caspases-3 through enzyme cutting was correct. A 894bp strap was observed by RT-PCR after pancreatic carcinoma cells being transfected with the pcDNA3.1(+)/r-caspases-3 by liposomes. No strap was found in control groups. A characteristic DNA ladder was observed in pancreatic carcinoma cells DNA elect r ophoresis, and transparent hypodiploid karyotype peak was found by FACS. Conclusion The plasmid of pcDNA3.1(+)/r-Caspase-3 was c onstructed successfully, the expression of r-Caspase-3 mRMA in pancreatic carc inoma cells was confirmed by RT-PCR, and pcDNA3.1(+)/r-Caspase-3 can induce a poptosis in pancreatic carcinoma cells.
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Objective To explore the value of clinical use of combined detection with tumor markers for pancreatic cancer. Methods Tumor markers CA242,CA19-9 and CA50 in serum of 32 patinets with pancreatic cancer;26 patients with non-pancreatic digestive tract cancers and 24 patietns with benign pancreatic or biliary tract diseases were measured by immunoradiometric assay (IRMA). Results The levels of three markers in serum and positive rates of patients with pancreatic cancer were higher than those of other patients. The effect of measurement combining CA242 with CA19-9 was the best. The sensitivity ,specificity and accuracy of diagnosis for pancreatic cancer were 92.6％, 73.8％ and 81.2％ respectively. The levels of CA242 and CA19-9 were positively relative to burden of pancreatic cancer, and serum levels of these two markers of patients with resectable pancreatic cancer were lower than those with unresectable, but on difference was observed for CA50. Conclusion Combined detection of serum CA242 and CA19-9 could prove the effectual indicator for finding the patients with pancreatic cancer in high risk population or for resectable pancreatic cancer. Pre-operative measurement of serum levels of CA242 and CA19-9 is helpful to evaluate the burden of the tumors and possiblity of resect for pancreatic cancers.
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Objective To explore the protective effect of estradiol preconditioning on the myocardium of isolated ischemic reperfused rats. Methods The instrument of Langendorff was used to establish the model of isolated ischemic reperfused heart and male rats were randomly divided into three groups: control, treatment and preconditioning groups. Results Compared with that of control and treatment groups, estradiol preconditioning decreased the release of LDH, CK and intracellular TnI more effectively. Conclusion Estradiol preconditioning produces more protective effect on the myocardium of isolated ischemic reperfused rats.