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.

Objective To explore the polymorphism at position G460W of ?-ADDUCIN and at position C825T of GNB3,and the genetic interaction between ?-ADDUCIN and GNB3 genes in a QiQihr essential hypetension population.Methods Three hundreds and thirty-one patients with EH and two hundreds and ninety-three healthy controls were enrolled.Genotyping was performed using PCR-RFLP technique.Results(1)genotype distributions of ?-ADDUCIN G460W(GG 0.177 vs 0.160,GW 0.580 vs 0.481,WW 0.242 vs 0.359,P=0.006) and GNB3(CC0.177 vs 0.353,CT 0.468 vs 0.541,TT 0.355 vs 0.106,P

Objective To investigate the correlation between the expression of STAT3 and MMP-2 in human pancreatic cancer,and to probe the mechanism by which STAT3 signal pathway regulates the expression of MMP-2 in pancreatic cancer cells.Methods Immunohistochemistry was used to detect the expression of STAT3,phosphorylated STAT3(p-STAT3)and MMP-2 in pancreatic cancer tissues of 34 cases and normal pancreatic tissues of 10 cases.Correlation between the expression of STAT3、p-STAT3 and MMP- 2 were statistically analyzed.Human pancreatic cancer cell lines SW1990 was cultured.AG490,an inhibitor of the upstream Janus kinase(JAK)of STAT3 was added into the culture medium.Electrophoretic mobility shift assay(EMSA)was used to detect STAT3 DNA-binding activity in SW1990 cells.Western blot was used to detect the expression of STAT3,p-STAT3 in SW1990 cells.In addition,the protein and mRNA expression of MMP-2 in SW1990 cells were determined by Western blot and RT-PCR,respectively. Results Immunohistochemistry revealed that the expression rate of STAT3,p-STAT3 were both higher in pancreatic cancer tissues than in normal pancreas tissues(P

Aim To investigate the protective effects of Diterpene Ginkgolides Meglumine Injection(DGMI)on SY5 Y cells damaged by oxygen-glucose deprivation and its functional mechanisms.Methods After 4 h of OGD,the cells were treated with 25 mg·L-1 drugs for 1 h.Subsequently,cell viabilities were measured by cell counting kit-8(CCK-8 kit)and cell apoptosis was measured by flow cytometric analysis.Furthermore, the mitochondrial membrane potential was detected by rhodamine123 staining.The levels of phospho-p38, phospho-p53,Bcl-2,Bax and cleaved caspase-9/3 were evaluated by western blot.Results DGMI signif-icantly increased the cell viabilities of SY5 Y cells dam-aged by OGD,and reduced OGD-elicited dissipation of mitochondrial membrane potential and cell apoptosis. Furthermore,DGMI also reduced p-p38,p-p53,Bax/Bcl-2 ratio,cleaved caspase-9 and cleaved caspase-3. Conclusion DGMI shows good neuroprotective effects on SY5 Y cells after oxygen-glucose deprivation.The underlying mechanisms may be associated with the sup-pression of p38/p53/Bcl-2 /caspase-9/caspase-3 sig-naling pathway.

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 .

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.

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.

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.

OBJECTIVETo investigate the clinical and laboratory features of very long chain acyl-CoA dehydrogenase deficiency ( VLCADD ) and the correlations between its genotype and phenotype.

METHODEleven patients diagnosed as VLCADD of Shanghai Jiaotong University School of Medicine seen from September 2006 to May 2014 were included. There were 9 boys and 2 girls, whose age was 2 d-17 years. Analysis was performed on clinical features, routine laboratory examination, and tandem mass spectrometry (MS-MS) , gas chromatography mass spectrometry (GC-MS) and genetic analysis were conducted.

RESULTAll cases had elevated levels of blood tetradecanoylcarnitine (C14:1) recognized as the characteristic biomarker for VLCADD. The eleven patients were classified into three groups: six cases in neonatal onset group, three in infancy onset group form patients and two in late onset group. Neonatal onset patients were characterized by hypoactivity, hypoglycemia shortly after birth. Infancy onset patients presented hepatomegaly and hypoglycemia in infancy. The two adolescent patients showed initial manifestations of exercise intolerance or rhabdomyolysis. Six of the eleven patients died at the age of 2-8 months, including four neonatal onset and two infant onset patients, with one or two null mutations. The other two neonatal onset patients were diagnosed since early birth through neonatal screening and their clinical manifestation are almost normal after treatments. Among 11 patients, seventeen different mutations in the ACADVL gene were identified, with a total mutation detection rate of 95.45% (21/22 alleles), including eleven reported mutations ( p. S22X, p. G43D, p. R511Q, p. W427X, p. A213T, p. C215R, p. G222R, p. R450H, p. R456H, c. 296-297delCA, c. 1605 + 1G > T) and six novel mutations (p. S72F, p. Q100X, p. M437T, p. D466Y, c. 1315delG insAC, IVS7 + 4 A > G). The p. R450H was the most frequent mutation identified in three alleles (13.63%, 3/22 alleles), followed by p. S22X and p. D466Y mutations which were detected in two alleles (9.09%, 2/22 alleles).

CONCLUSIONThe ACADVL gene mutations were heterozygous in our patients. The mortality of neonatal onset form and infant onset form is much higher than the late onset form patients, suggesting a certain correlation between the genotype and phenotype was found. The earlier diagnosis and treatment of VLCADD are of vital importance for the improvement of the prognosis of the patients.

Acyl-CoA Dehydrogenase, Long-Chain ; deficiency ; genetics ; Adolescent ; Age of Onset ; Alleles ; Asian Continental Ancestry Group ; Child ; Child, Preschool ; China ; Female ; Genetic Testing ; Genotype ; Heterozygote ; Humans ; Infant ; Infant, Newborn ; Lipid Metabolism, Inborn Errors ; complications ; genetics ; Male ; Mitochondrial Diseases ; complications ; genetics ; Muscular Diseases ; complications ; genetics ; Mutation ; Neonatal Screening ; Phenotype ; Prognosis ; Rhabdomyolysis ; etiology ; Spectrum Analysis ; Tandem Mass Spectrometry

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.