Objective To investigate the expression ot XAF1 protein in pancreauc cancer,and its relationship with clinicopathological parameters and prognosis.Methods The tissue microarray was composed of 89 pancreatic cancer and 21 normal pancreatic tissues,and immunohistochemistry was used to examine the expression of XAF1 protein.The correlation between XAF1 expression and clinicopathological parameters was analyzed.The 89 patients were followed,and the survival was shown in Kaplan Meier curve.Cox proportional hazards regression was used to identify risk factors for the survival of pancreatic cancer patients.Results The negative expression rate of XAF1 was 44.9％ (40/89),and it was 9.5％ (2/21) in normal tissue,and the difference between the two groups was statistically significant (P ＜0.05).The expression of XAF1 was negatively associated with tumor staging (P ＜ 0.05),but it was not associated with age,gender,lymph node metastasis,distant metastasis,pathological grading,surgical margins,vascular and neural invasion.Patients with negative expression of XAF1 had much shorter survival than patients with positive expression.Univariate analysis showed that negative expression of XAF1,pathological grading,lymph node metastasis,distant metastasis was associated with the survival (P ＜ 0.05),and the multivariate analysis indicated that negative expression of XAF1 was an independent prognostic factor for survival of pancreatic cancer.Conclusions XAF1 may be involved in the development and growth of pancreatic cancer,and it is related with patient's prognosis.

Objective To determine the serum KL-6 level in patients with pancreatic carcinoma and investigate its diagnostic value. Methods The data of 53 pancreatic carcinoma (PC) patients; 68 chronic pancreatitis (CP) patients and 51 patients with high risks of pancreatic cancer (HR) with complete follow-up data were studied, and 50 healthy volunteers were used as controls. ELISA was used to measure the serum levels of KL-6, MUC4, and CA50. Radioimmunoassay methods were used to measure the serum CA19-9 levels. The sensitivity and specificity of KL-6 for the diagnosis of PC were determined, and the relationship between its levels and clinicopathologic parameters, patients' outcome were analyzed. Results The serum KL-6 levels were (753±548), (135±93), (105±55) and (99±50) U/ml in PC group, CP group, HR group and control group, and the value in PC group was significantly higher than those in other 3 groups (P <0.01). Using >232 U/ml as the cut-off point, the sensitivity and specificity of KL-6 for the diagnosis of PC was 96% and 94%. Using >244 U/ml as the cut-off point, the sensitivity and specificity of KL-6 for the diagnosis of PC was 97% and 91%. The clinical value of KL-6 was higher than those of CA19-9, CASO and MUC4. The serum level of KL-6 was not associated with clinicopathologic parameters. The median survival of patients with serum level of KL-6 =300 U/ml was (9.3 ±1.2) months, which was significantly longer than that in patients with serum level of KL-6 >300 U/ml [ (4.6 ±0.7) months, P =0.006]. Conclusions KL-6 might be a useful serological marker of pancreatic cancer, and it may play a role in the differential diagnosis between pancreatic cancer and chronic pancreatitis.

Objective To investigate the effect of inhibiting the expression of Navl. 8 sodium channels by an antisense oligodeoxynudeotide(ODN) on visceral hypersensitivity. Methods The visceral hypersensitivity animal models induced by giving neonatal Sprague-Dawley rats colorectal distention(CRD) after postnatal days 8, 10 and 12. Animals were injected intrathecally either Navl. 8 antisense ODN (research group) or mismatch ODN (control group) twice a day for 3 days at the 8th week. After that, the expression of Navl. 8 in each group was examined by RT-PCR, and abdomiral withdrawal reflex(AWR) score and spinal c-Fos expression were evaluated to assess whether the intervention can relieve the visceral hypersensitivity. Results After intrathecal injection with the antisense ODN, the expression of Navl. 8 in each group decreased.AWR score and the number of c-Fos positive neurons in the spinal cord, especially in the area 1 and 3, decreased in the model group, while they did not change in the control group. Conclusions Interventing the expression of Nav1.8 may relieve the visceral hypersensitivity in this model of irritable bowel syndrome.

Objective To investigate the effect of tumor necrosis factor receptor-p55/p75(TNFR-p55/ TNFR-p75) on the pathogenesis of severe acute pancreatitis (SAP) in rats. Methods Thirty-six male Sprague-Dawley rats were divided into 2 groups: SAP group and control group. SAP model was ~induced by injection of 5% sterile sodium taurocholate into biliopancreatic duct, while control group was only given sham operation. Rats were sacrificed at 3, 6, and 12 hours after the onset of operation. Blood sample and pancreatic tissues were collected. The severity of pancreatitis was assessed according to the level of serum amylase and histological scoring. The serum levels of tumor necrosis factor-?(TNF-?) were examined by ELISA. Expressions of TNFR-p55 mRNA and ~TNFR-p75 mRNA in pancreatic tissues and peripheral blood mononuclear cells (PBMC) were measured by semi-quantitative RT-PCR. Results The levels of serum amylase and TNF-? in SAP group were both significantly higher than those in the control group at each time point (P

Objective To look for and confirm the downstream regulated gene of microRNA (miRNA)-148a in pancreatic cancer cell line.Methods The target gene regulated by miRNA-148a was predicted through bio-informatics analysis.The plasmid containing desired gene and with luciferase 3'-untranslated region (3'-UTR) reporter was constructed.Pancreatic cancer cells BXPC-3 were transfected with analogs and inhibitors of miRNA-148a by liposomes.The activity of luciferase was measured to determine whether miRNA-148a directly connected with desired gene.The expression level of miRNA-148a was changed in BXPC-3 cells,and the changes of target gene v-verb-b2 erythroblastic leukemia viral oncogene homolog 3 (awian) (ErbB3) expression were detected by Western blot at protein level.The data were analyzed by one way ANOVA.Results There was a conservative binding site of ErbB3 with miRNA-148a detected by bio-informatics analysis,miRNA-148a directly combined with ErbB3 and the activity of luciferase decreased to (25.00+47.00) ％ of the negative control (F=4.66,P＜ 0.01).After miRNA-148a overexpression,the gray value of ErbB3 expression in BXPC-3 cells decreased to (26.16±4.69)％ of control group (F=6.563,P＜0.05).Conclusion miRNA-148a directly targeted and regulated the expression of ErbB3 in pancreatic cancer cell line BXPC-3.

Objective To investigate the role of connexin 43(Cx43)in the apoptosis of pancreatic cancer cell line BxPC and its possible mechanism.Methods pcDNA-Cx43,pcDNA-Cx43N,pcDNA3.0,siRNA-Cx43 and siRNA-NC were transfected into BxPC3 cells via liposome method.Cx43 protein and Cytochrome C(Cyt C)concentration was determined by Western blot,and the apoptosis was analyzed by Annexin V/PI binding assay.The mitechondria apoptosis pathway involved in Cx43 associated apoptosis was examined which contains the depolarization of mitechondrial membrane potential (MMP);fluorospectrophotometer was used to measure the activities of caspase-3 and caspase-9. Gap junction intercellular communication(GJIC) was determined by dye-transfer method.Results Cx43 protein expression increased after BxPC3 transfeetion,apoptosis rate increased from(6.35±0.43)%in empty vector transfection group to(14.29±1.24)%;after H202 treatment,apoptosis rate increased from(20.34±2.47)%to(31.27±2.56)%(P<0.05).Meanwhile,mitochondrial membrane potential was decreased,Cyt C was increasingly released from mitochondria,caspases activities were increased;after siRNA43 interference,apoptosis rate decreased from(7.42±0.47)% to(5.19±1.37)%,after H_2O_2 treatment,apoptosis rate decreased from (19.43±1.71)%to(11.67±1.97)%(P<0.05).Decreased mitochondrial membrane potential and Cyt C release were observed,caspases activities were decreased.GJIC of pcDNA-Cx 43 transfection group increased from 14.52±0.57 to 23.05±3.84.and it increased from 1.70 ±0.24 to 3.84 ±0.45 in the presence of β-GA(P<0.05).But the apoptosis rate was not significantly different.Conclusions Cx43 could promote BxPC3 apoptosis via mitochondrial apoptotic signal pathway,and the possible mechanism included signal pathway other than GJIC.

Objective To analyze the relationship between blood glucose level, blood pressure and weight with pancreatic cancer genesis. Then to explore the metabolism associated risk factors in pancreatic cancer genesis. Methods Form December 2002 to September 2009 in Ruijin Hospital, 548 pancreatic cancers with pathology diagnosis after pancreatectomy were collected for the study with retrospective analysis method. The association of pancreatic cancer with blood glucose level, blood pressure, weight and other metabolic factors were analyzed. Results With principal component analysis, it suggested that there were strong correlation between blood glucose level, blood pressure and weight index (BMI) increasing with pancreatic cancer. The contribution rates were 3. 614%,25. 236%, 15. 418% and 12. 918%, respectively. Single factor analysis indicated that the association between pancreatic cancers and new onset diabetes mellitus (duration≤ 2 years) was stronger than that of long-term diabetes mellitus. The occurrence rate of pancreatic cancer in patients with long-term diabetes whose blood glucose level was not well controlled recently while well controlled previously (44.6 % ) was significant hister than that in patients without diabetes (5. 6% , P＜0.05). The fasting blood glucose level of these PC patients ( 13.87± 3. 49 mmol/L) was significantly higher than new onset and other long-term diabetes patients, the comparative risk was 13.46 (95% CI 4. 560,39. 731). BMI increasing was a risk factor of pancreatic cancer, but there was no significant statistical difference between risk degree and BMI increasing level. All above metabolic diseases were risk factors of pancreatic cancer, but for pathology, location and stage of pancreatic cancer there was no statistical difference in theses factors. Conclusion This study suggested diabetes, BMI increasing and hypertension were high risk factors of pancreatic cancer genesis. New onset and long-term diabetes patients whose blood glucose not controlled well recently should be watched carefully for pancreatic cancer. Early treatment and intensive follow-up of metabolic disease might be helpful to early diagnosis and prognosis of pancreatic cancer.

Objective To investigate the relationship between the activity of Janus kinase/signal transducer and activator of transcription (JAK/STAT) signaling pathway in cerulein-induced pancreatic acinar cell line AR42J. Methods The in vivo model of AP was induced by cerulean treated pancreatic acinar cell line AR42J, then RPM and AG490 were given for intervention. Western blot was used to determine theexpressions of JAK1 and phosphorylation JAK1 ( P JAK1 ) , STAT1, PSTAT1 and TNFα, IL-1β, IL-6. The expressions of IL-6, IL-1 β, and TNFα mRNA were measured by RT-PCR. Survival rate of cells was evaluated by trypan blue stain. Results The relative expressions of JAK1, P JAK1, STAT1, P STAT1 and TNF-o, IL-1β, IL 6 without cerulean treatment were 0.09 ±0.04,0.14 ±0.08,0.21 ±0.09,0.12 ±0.12,0.10 ±0.02,0.08 ± 0.03,0.02 ± 0.02. After cerulean treatment, the expressions of abovementioned protein increased in a time-dependant manner, the expressions at 24h were 0.53 ± 0.09,0.53 ± 0.13,0.56 ± 0.09,0.55 ± 0.10,0.25 ± 0.04,0.25 ±0.09,0.27 ±0.07, which were significantly higher than those in the control group (P ＜0.05). 2 4 h after RPM and AG 4 9 0 inhibition, the expressions of TNF-α, IL-1 β, IL-6 proteins significantly decreased to 0.17 ± 0.03and 0.17 ± 0.01,0.15 ± 0.05 and 0.14 ± 0.07,0.19 ± 0.04 and 0.19 ± 0.05; their expressions of mRNA significantly decreased ( P ＜ 0.05 ). The cell survival rates in RPM and AG490 treatment group were (72.4 ± 11.2) %, (69.7 ± 9.8 ) %, and in cerulein-stimulated cells (42.2 ± 12.3 ) % ( P ＜ 0.05 ).Conclusions The JAK1/STAT1 signaling pathway was involved in pancreatic inflammatory response with cerulein stimulation. Early treatment with inhibitors to the JAK1/STAT1 signaling pathway might control the inflammatory response in acute pancreatitis.

Aim To study the anti-inflammatory activity of the Channa argus bile.Methods The bile was isolated and purified by extraction and silica gel column chromatography.Then the compounds were identified by hydrogen and carbon spectra.The spleen lymphocytes proliferation assay and Lipopolysaccharide(LPS) induced mouse macrophage RAW264.7 releasing Nitrogen Monoxide(NO) experiment were used to evaluate the anti-inflammatory activity.Results Compound(C1) of sodium taurocholate and compound(C2) of sodium taurochenodeoxycholate were isolated by activity tracing.The cell relative viabilities of the two compounds on Concanavalin A(Con A) induced spleen lymphocytes proliferation assay were 65.9%±11.7% and 60.5%±9.4%, which were significantly different from the result of model group (P<0.01), respectively.The NO production of LPS-induced RAW264.7 release of NO was (16.4±1.9) μmol·L-1 and (15.5±1.7) μmol·L-1, which were significantly different from the result of model group(P<0.01).Conclusion Sodium taurocholate and sodium taurochenodeoxycholate from Channa argus perform the anti-inflammatory activities but have no cytotoxic effect on spleen lymphocytes and macrophage.

Objective To investigate the circulating microRNAs carrier in pancreatic cancer.Methods Pancreatic cancer cell lines SW1990 and BxPC3 were routinely cultured and serum of 6 patients with pancreatic cancer and 6 healthy subjects (control group) were collected.Serum and pancreatic cancer cell line supernatant microvesicles (MV) were obtained by gradient centrifugation.The expression of Ago2,CD63 was detected by Western blotting,the expression of microRNA in microvesicle section and microvesicle-free section in serum was detected by using quantitative PCR method.Results The supernatant MV of pancreatic cancer cell lines expressed Ago2,CD63 protein,and these MV carried different microRNAs in different cell lines.In the serum of pancreatic cancer and control group,miR-20a,miR-21,miR-24,miR-25,miR-191,miR-483-5p were detected,but the quantity was relatively higher in MV section,and the expression of microRNAs in pancreatic cancer's MV was inconsistent with that of control group.The expression of miR-20a,miR-24,miR-191 in pancreatic cancer group was (2.93 ± 0.29),(2.73 ± 0.46),(2.39 ± 0.51) times as much as those in control group,and the difference between the two groups was statistically significant (F =75.97,25.80,12.94,P ＜ 0.05 or ＜ 0.01).Conclusions The main circulating microRNAs carrier in pancreatic cancer is microvesicle.