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Objective To investigate the expression of miRNA-301a-3p in pancreatic cancer and to correlate the expression on invasion , migration and colony formation of pancreatic cancer cells .Methods The expression of miRNA-301a-3p in 20 paired pancreatic cancer tissues and matched adjacent tissues , and pancreatic cancer cell lines and normal pancreatic ductal cells were detected by real -time PCR.miRNA-301a-3p mimics or inhibitors were used to up-regulate or down-regulate the miRNA-301a-3p level in pancre-atic cancer cell lines in order to figure out the effects of miRNA-301a-3p on cell invasion, migration and col-ony formation of pancreatic cancer cells , respectively .Results In pancreatic cancer tissues and cell lines , miRNA-301a-3p was significantly up-regulated when compared with the matched adjacent tissues ( P <0.05) and normal pancreatic ductal cells (P<0.05), respectively.Overexpression or downexpression of miRNA-301a-3p enhanced or suppressed colony formation , invasion and migration abilities of pancreatic cancer cells in vitro.Upregulation of miRNA-301a-3p promoted tumorigenesis in vivo.Conclusion miR-NA-301a-3p might function as an oncogene to promote tumorigenesis in pancreatic cancer .
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Objective To study the relationship between the expression of Caveolin-1 (Cav-1) and epithelial-mesenchymal transition(EMT) or metastasis in human pancreatic cancer.Methods The Cav-1 protein and EMT markers in highly metastatic pancreatic cancer cell lines (L3.7,Panc02-H7) and poorly metastatic pancreatic cancer cell lines (COLO357,Panc02) was detected by Western blotting and immunofluorescence.The morphology of the pancreatic cancer cells were observed under phase-contrast photomicrographs.The plasmids pcDNA3.1-caveolin-1 and control vector pcDNA3.1 were transfected into COLO357 cells by Lipofectamine LTX.The expression of Cav-1 protein,E-cadherin and vimentin were detected,and the morphology of COLO357 cells observed.Results L3.7 and Panc02-H7 cells had strong positive Cav-1 staining and high expression of mesenchymal marker (vimentin,N-cadherin) and low epithelial marker (E-cadherin,β-catenin),whereas COLO357 and Panc02 cells with typical epithelial morphology were weak positive in Cav-1 staining and of low expression of vimentin,N-cadherin and high expression of E-cadherin,β-catenin.In Cav-1 transfected COLO357 cells vimentin expression increased,and E-cadherins decreased resulting in typical morphology changes of EMT.Conclusions Overexpression of caveolin-1 is correlated with epithelial-mesenchymal transition of pancreatic cancer cells and cancer metastasis.
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Objective To investigate the effect and mechanism of RNAi-mediated STAT3 gene silencing on epithelial-to-mesenchymal transition (EMT) of human pancreatic cancer cells.Methods Lentivirus vector mediating RNA interference targeting STAT3 was constructed in SW1990 cell line.The invasion ability of SW1990 cells was determined by cell invasion assay in vitro.Cell proliferation and cell cycle of SW1990 cells were also detected.The expression of EMT related genes such as STAT3,P-STAT3,Twist,Snail and E-cadherin were analyzed by reverse transcription-PCR,real-time PCR,and Western blotting.Results Silencing of STAT3 with RNAi not only markedly reduced proliferation but also greatly decreased the invasion ability of SW1990 cells.The mRNA level and protein expression of Snail decreased significantly (P ＜ 0.05),but those of E-cadherin increased significantly (P ＜ 0.05),compared to parental cells.However,no difference was on the expression of Twist in SW1990 cell line.Conclusions STAT3 signaling pathway plays an important role in the process of EMT.Silencing of STAT3 with RNAi can significantly inhibit EMT by downregulating expression of Snail and E-cadherin in pancreatic cancer cells.
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Objective To investigate the effects and mechanism of IL-6 on the epithelial to mesenchymal transition of human pancreatic cancer cells.Methods IL-6 was added into the culture media of human pancreatic cancer cells Capan-2,SW1990,and STAT3-siRNA-SW1990.Cell growth was measured by MTT assays.STAT3,p-STAT3,Snail,Twist,and E-cadherin mRNA and protein expression were examined using real-time fluorescence quantitative polymerase chain reaction (RT-PCR)and Western blot,respectively.The invasion abilities of SW1990 and Capan-2 cells were determined by a cell invasion assay in vitro.Results Our results showed that 100 μg/L of IL-6 significantly promoted the growth and invasion abilities of Capan-2 and SW1990 cells (P＜0.05).The use of IL-6 not only markedly increased the protein expression of P-STAT3 and Snail,but also greatly decreased the mRNA and protein expression of E-cadherin.The use of IL-6 can not change the mRNA and protein expression of Snail and E-cadherin.Conclusion Activation of the STAT3 signal transducer pathway with IL-6 can promote the epithelial to mesenchymal transition of pancreatic cancer cells in vitro through up-regulation of Snail and down-regulation of E-cadherin expression.Therefore the STAT3 signal transducer may provide a novel therapeutic target for the treatment of pancreatic cancer.
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ObjectiveTo evaluate the clinical efficiency and safety of cefmetazon in the prevention Department of General Surgery,First People's Hospital,Shanghai Jiaotong University,Shanghai 200080,Chinaand or treatment of infections in general surgery. MethodsA multicenter,prospective and open-labeled trial was conducted. In the prevention group,1700 patients were enrolled in clean-infection surgery,cefmetazon was given 1 g iv half an hour before the surgery started,and 1 g iv twice daily after the surgery for 3 days.Clinical response was evaluated in terms of both cure ( disappearance of pre treatment symptoms)and pathogen. In the treatment group,897 patients were diagnosed as peritonitis, cholecystitis and cholangitis,the patients were given cefmetazon 2 g iv twice a day for 7 - 14 days,clinical response and microbiological efficacy were assessed.ResultsIn prophylactic group,1449 patients were finally included.The clinical efficacy was 100％ (1449/1449).In the treatment group,a total of 897 patients were enrolled,and 110 patients failed for assessment of clinical efficacy,787 patients were included in the PPS population,the clinical efficacy was 90.7％ (714/787); Bacterial eradication rate was 92％ (46/50).Adverse reaction rates in prevention group and treatment group were 1.3％ (22/1700) and 1.2％ (11/897),including mild nausea and vomitting.ConclusionsCefmetazon is effective and safe in prevention and treatment of Postoperative infections in general surgery.
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Objective To screen the genes related with signal transducers and activators of transcription 3 (STAT3) regulating pancreatic cancer invasion and metastasis by gene chips.Methods Human pancreatic cancer cell line SW1990 stably expressing low level of Stat3 was established by lentivirus transfection,while cells transfected with mock plasmid and cells without transfection served as control groups.The differences of invasion and metastasis related genes expression among the three groups were screened by gene chips.STAT3 mRNA and protein expression was measured by real-time PCR and Western blot.Three differentially expressed genes (MMP-7,IL-1β and IgTα7) were verified.ResultsThe expression level of STAT3 mRNA was 0.391 ± 0.037 after pancreatic cancer SW1990 cell trarsfected with STAT3 targeted lentivirus,which was significantly lower than those in mock plasmid group (1.002 ± 0.015) and nontransfected group ( 1.206 ± 0.042,P ＜ 0.05 ) ; the expression level of STAT3 protein was 182.38 ± 65.32,which was significantly lower than those in mock plasmid group (223.40 ±58.40) and non-transfected group (212.33 ±53.69).Eight invasion and metastasis related genes of SW1990 lowly expressing Stat3 were upregulated,while 3 genes were down-regulated.By verification,the mRNA level of MMP-7 and IL-1β were lower than in control group transfected with mook plassmid(0.287 ± 0.115 vs 1.010 ± 0.124,t =19.45,P =0.000;0.490 ± 0.10 vs 1.002 ± 0.002,t =13.83,P =0.000),but the mRNA level of IgTα7 was not decreased (1.173 ±0.280 vs 0.998 ±0.003,t =4.236,P =0.094).Meanwhile,the protein level of MMP-7 was significantly down-regulated when Stat3 was knocked down.ConclusionsStat3 causes changes of expressions of many invasion and metastasis-related genes of SW1990,and MMP-7 may be the main target gene regulated by Stat3.
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Objective To investigate the expression and prognostic significance of CD151, c-Met and integrin alpha 3, alpha6 in pancreatic ductal adenocarcinoma (PDAC). Methods The expression of CD151, c-Met and integrin alpha3, alpha6 in 71 patients with PDAC and 10 samples of normal pancreas tissues were detected by immunohistochemistry, and the relationship between the expression of CD151, c-Met and integrin alpha 3, alpha 6 and the clinicopathological features, prognosis of these patients was analyzed. Results The positive expression rates of CD151, c-Met and integrin alpha 3, alpha 6 in PDAC were 81.69% (58/71) , 69.01% (49/71), 69.01% (49/71) and 84.51% (60/71) , and there was no expression in normal pancreas tissues. The expressions of CD151, c-Met were significantly associated with TNM stage and lymph node metastasis (P < 0.05). The expression of CD151 was positively correlated with the expressions of c-Met and integrin alpha3, alpha6 (r =0.583, P =0.000, r = 0.457;P =0.000, r = 0.671 ;P =0.000). Univariate analysis suggested the expression of CD151, c-Met and integrin alpha3, alpha6 was associated with survival (P<0.05). Multivariate analysis suggested the expression of CD151, c-Met was the independent prognostic factor for post-operative survival. Conclusions CD151, c-Met and integrin alpha3, alpha6 play a role in the development, metastasis and prognosis of PDAC, and they might be new markers to predict biological behavior and the prognosis of PDAC patients.
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Stat3 is a member of the STAT (signal transducer and activator of transcription) protein family. It can be activated by many cytokines, growth factors and carcinogens. Activation of Stat3 in the cytoplasm leads to its dimerization, translocation into the nucleus, DNA binding and gene transcription. Constitutively activated Stat3 is associated with cellular transformation, invasion, metastasis, angiogenesis, chemoresistance and radioresistance, and therefore suppress apoptosis of cancer.
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Objective To investigate the effect and mechanism of RNAi-mediated STAT3 gene silence on human pancreatic cancer cells growth in vivo. Methods STAT3 shRNA expression vector was stably transfected to SW1990 cells. STAT3 and p-STAT3 protein was examined using Western blot. The growth ability of SW1990 cells in vivo was determined in a subcutaneous tumor model of nude mice. Western blot was performed to detect the protein expression of Bcl-xL and cyclin D1. Results The protein expression of STAT3 and p-STAT3 decreased by 90% and 92% by stable transfection of STAT3 shRNA expressing vectors(P ＜0. 05). Inhibition of STAT3 with RNAi significantly inhibited the growth ability of SW1990 cells in vivo( P ＜ 0. 05 ). The tumor weight significantly decreased( P ＜ 0. 05 ). Moreover, the relative Bcl-xL and cyclinD1 protein expression in SW1990-RNAi cells reduced by 56% and 50% compared with that of the parental SW1990 cells, respectively (P ＜ 0. 05). Conclusions Inhibition of STAT3 with RNAi significantly inhibits the growth ability of pancreatic cancer cells through down-regulating Bcl-xL and cyclin D1.
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Objective To investigate the expression of HIF-1α and P-gp protein in pancreatic carcinoma and determine their clinicopathological significance and the correlation between the expression of HIF-1α, P-gp and the clinical prognosis. Method In samples from 74 cases of pancreatic carcinoma and 10 healthy individuals, the expression of HIF-1α and P-gp were detected by immunohistochemical method. Results The positive expression rate of HIF-1α and P-gp was 75.7% and 86.5%,respectively, which were remarkably higher than that in normal pancreatic tissue (P＜0.05). There was a positive correlation between the expression of HIF-1α and that of P-gp. The aberrant expression of HIF-1α and P-gp was associated with lymph node metastasis but not the location, size, clinical stages and nerve invasion of the tumor. Patients with high intensity of HIF-1α and P-gp expression showed a significantly lower median survival time than those with low intensity expression.Conclusions The expression of HIF-1α and P-gp is up-regulated in pancreatic carcinoma and there is a positive correlation between them. The expression of HIF-1α and P-gp might be related to the lymph node metastasis and poor prognosis.
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Objective To investigate the effects and mechanism of IL-6 on invasion and metastasis of human pancreatic cancer cells. Methods IL-6 was added into the culture media of human pancreatic cancer cells Capan-2 and SW1990. Cell growth was measured by MTT assay. Western blot and immunocytochemistry were performed to detect Phosphorylated STAT3 (P-STAT3) protein. VEGF and MMP-2 mRNA and protein expression were examined using fluorescence quantitative polymerase chain reaction (RT-PCR) and Western blot, respectively. The invasion ability of SW1990 and Capan2 cells was determined by cell invasion assay in vitro. Results 100 ng/mL IL-6 significantly promoted growth and invasion ability of Capan-2 and SW1990 cells (P＜0.05). The use of IL-6 not only markedly increased the protein expression of P-STAT3, VEGF and MMP-2, but also greatly increased the mRNA expression of MMP-2 and VEGF. Conclusions STAT3 signal transducer pathway activation with IL-6 can promote the invasion ability of pancreatic cancer cells in vitro through up-regulation of MMP-2 and VEGF expression. STAT3 signal transducer may provide a novel therapeutic target for the treatment of pancreatic cancer.
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Objective To investigate the expression of Akt and phosphoryled Akt (p-Akt1) protein in pancreatic carcinoma and to determine the clinical significance. Methods In 74 cases of pancreatic carcinoma and 10 cases of normal pancreatic tissue samples, the expression of Akt and p-Akt1 were detected by immunohistochemical method, and the its relationship with clinicopathologic characteristics and prognosis were analyzed. Results The positive expression rate of Akt and p-Akt1 in pancreatic carcinoma were 87.8% and 83.8% respectively, while there was no expression of Akt and p-Akt1 in normal pancreatic tissue, and the difference was statistically significant (p < 0.05). There was a positive correlation between the expression of Akt and p-Akt1 in pancreatic carcinoma (r =0.274, P =0. 018). The expression of Akt and p-Akt1 was not significantly associated with the age, sex, location, size, pathology stages, lymph nodes metastasis, clinical stages and nerve invasion of the tumor (P >0.05). But the higher expression of p-Akt1 was associated with T stages and TNM staging (p =0. 002). Patients with high intensity of Akt and p-Akt1 expression showed a significantly longer median survival time [(16.0 ± 5.7) month and (23.0 ± 5.5) month, respectively]than those with low intensity expression [(9.3 ± 0.2) month and (11.1 ± 1.8) month (P = 0. 007 and P = 0.004) respectively]. Conclusions p-Akt1 expression is a significant positive prognostic factor for pancreaticcarcinoma and detection of p-Akt1 expression may be of clinical value.
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Objective To investigate the expression of E-cadherin and Snail proteins in rectal cancer and their significance. Methods The expression of Snail and E-cadherin proteins was detected using immunohistochemical SABC method in 101 cases of rectal cancer tissues. Results The positive rate of Snail in rectal cancer was 78.2% (79/101). The negative expression rate of E-cadherin in rectal cancer was 62.4% (63/101). The expression of Snail and E-cadherin were significantly related with the lymph node metastasis and Dukes' stage of rectal cancer (P ＜ 0.05). Conclusion The overexpression of Snail and the decreased expression of E-cadherin might be important biological markers for malignant transformation, invasion and metastasis of rectal carcinoma.
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Loss of epithelial characteristics and acquisition of a mesenchymal phenotype are the main features of epithelial-mesenchymal transition (EMT). EMT plays an important role in tumor invasion and metastasis through increasing cell migration, invasion and anti-apoptotic capacity. Several mechanisms involved in EMT regulate invasion and metastasis of pancreatic cancer,including cadherin switch, growth factors, transcription factors, microRNA and signaling pathways.
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Pancreatic cancer is a frequent malignant tumor in digestive tract with extremely poor prognosis,characterized by difficult early diagnosis,high malignancy,poor resective,limited response to chemotherapy and radiotherapy and an intense fibrotic reaction known as tumor desmoplasia.Pancreatic stellate cells play an important role in this reaction and can stimulate pancreatic cancer cells proliferation,invasion and metastasis through the interaction with pancreatic cancer cells.This review describes the role of pancreatic stellate cells in the process of pancreatic cancer progression.
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Objective To investigate the effect of interleukin 6 (IL-6) on the growth and proliferation of human pancreatic cancer cell line Capan-2 and the signal transduction pathway. Methods MTr method was used to detect the effect of IL-6 of different concentrations on the growth and proliferation of Capan - 2 cells; cell apoptosis was detected by flow cytometry; the intracellular localization of phosphorylated STAT3 (P-STAT3) was determined by immunocytochemistry and western blot were used to detect P-STAT3, bcl-xl and Cyclin D1 in Capan-2 cells stimulated by IL-6. Results IL-6 (100 ng/ml) could remarkably promote the growth of Capan-2 cells from 1 to 4. 965 ± 0. 18 (P < 0. 05) ; the percentage of apoptosis decreased significantly from (3.21 ±0.23)% to (1.98 ±0.67)% (P <0.05) ; the expressions of P-STAT3, bcl-xl, Cyelin D1 increased significantly (P < 0.05), and the expressions of bcl-xl was positively correlated with that of P-STAT3 (r =0.985, P =0.015) ; the expressions of Cyclin DI was also positively correlated with that of P-STAT3(r=0.914,P=0.036),Conclusions IL-6 activate JAK/STAT signal transduction pathway,which played an important role in the growth and proliferation of Capan-2 cells in the presence of IL-6.
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Objective: In order to investigate the effects of activating and blocking Stat3 signaling pathway on invasion ability of human pancreatic cancer cells and explore its action mechanism.Methods:Human pancreatic cancer Capan-2 cells were treated with IL-6. SW1990 human pancreatic cancer cells were treated with AG490. Cell proliferation was measured by MTT assay. Western blotting and immunocytochemistry were performed to detect expression of phosphorylated Stat3 (p-Stat3) protein. Real-time fluorogentic quantitative PCR (RFQ-PCR) and Western blotting were used to detect the mRNA and protein expression of VEGF and MMP-2 mRNA, respectively. The invasion abilities of SW1990 and Capan-2 cells were determined by cell invasion assay in vitro. Results:IL-6 stimulated the proliferation of Capan-2 cells (P<0.05), elevated the expression of p-Stat3, increased the mRNA and protein expressions of vascular endothelial growth factor (VEGF) and matrix metalloproteinase 2 (MMP-2) (P<0.05), and enhanced the invasion ability of Capan-2 cells. AG490 inhibited the proliferation of SW1990 cells (P<0.05), down-regulated the expression of p-Stat3, markedly decreased the mRNA and protein expression of VEGF and MMP-2 (P<0.05), and weakened the invasion ability of SW1990 cells. Conclusion:Stat 3 signaling pathway plays an important role in the invasion and metastasis of pancreatic cancer. Stat 3 signaling transduction pathway may provide a novel therapeutic target for the treatment of pancreatic cancer.
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Objective To investigate the effect of RNAi-mediated sTAT3 gene silencing on the invasiveness of human pancreatic cancer cells. Methods STAT3 shRNA expression vector was stably transfected to SW1990 cells.STAT3 mRNA and protein expression were examined using reverse transcription polymerase chain reaction(RT-PCR)and Western blot,respectively.The invasion ability of SW1990 cells was determined by cell invasion assay in vitro.RT-PCR and Westem blot were performed to detect the mRNA and protein expression of the MMP-2 and VEGF,respectively. Results mRNA and protein expression of STAT3 were inhibited significantly by stable transfection of STAT3 shRNA expressing vectors.STAT3 silence with RNAi significantly inhibited the invasion ability of SW1990 cells decreasing protein and mRNA expression of MMP-2 and VEGF in SWl990 cells. Conciusion STAT3 silence with RNAi significantly inhibits the invasion ability of pancreatic cancer cells through down-regulating MMP-2 and VEGF.
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Background and purpose:Micro-environmental hypoxia is a common phenomenon in most human solid tumors,and this investigation is done to observe the expression of HIF-1? and chemo-resistance-associated genes in human colon cancer cell line under hypoxic micro-environment in vitro,and study the influence of micro-environmental hypoxia on chemo-resistance and the possible mechanisms in human colon cancer.Methods:Human colon cancer cell line SW620 was cultured under hypoxia for 12,24,48 hr,with normoxia as control.Then the expression of HIF-1? and chemo-resistance-associated genes mdr1/P-Gp、LRP were investigated by RT-PCR and western-blot.Results:With prolongation of the hypoxic time,the mRNA expressions of HIF-1? and LRP remained at the same level,but the mRNA expressions of mdr1 showed a time-dependent increase(P
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Objective To investigate the indications and results of combined liver-kidney transplantation.Methods From Jan. 2001 to Dec. 2003, 15 patients were subjected to combined liver-kidney transplantation in our department. The underlying diseases included hepatitis B viral cirrhosis complicated by HRS ( n= 8), hepatitis B viral cirrhosis complicated by uremia ( n =2), hepatitis B viral cirrhosis complicated by diabetic nephropathy ( n =1), polycystic liver and kidney disease ( n =2), Caroli's disease and polycystic kidney ( n =1), alcoholic liver cirrhosis complicated by uremia ( n =1). The surgical procedure, perioperative complications, acute and chronic rejection, the recurrence of hepatic viral B hepatitis, and the result of follow-up were analyzed.Results The graft function in 15 cases of combined liver-kidney transplantation was restored well after operation. The 6-month and one-year survival rate was 100%. One patient was supported by respiration machine for 48 days. The complications occurred in 3 patients after operation, including one case of gastroenternal bleeding repeatedly and one case of postoperative wound bleeding subject to non-surgical treatment, and one case of stenosis of biliary anastomosis subject to ERCP. Only one patient experienced a rejection episode of the liver. No acute rejection of the kidney graft occurred. One patient was died from liver graft function failure by recurrence of hepatitis B after 30 months.Conclusions Combined liver-kidney transplantation is only radical treatment method for patients with end-stage liver disease with chronic renal dysfunction or chronic renal failure. In the patients with hepatitis B,lamividine and hepatitis B immunoglobin can prevent the recurrence of hepatitis B.