Objective To investigate the application value of pancreatic duct guide wire and transpancreatic septotomy with precutting technique in difficult endoscopic retrograde cholangiopancreatography.Method258 patients who underwent difficult endoscopic retrograde cholangiopancreatography from April 2014 to April 2016 were selected as study subject, 128 patients among them received the technique of pancreatic duct guide wire, the other 130 patients received transpancreatic septotomy with precutting techniques. The success rate, intubation time and incidence of complications were compared between these two methods for cannulation.Result There was no signiifcant difference in preoperative clinical data between the two groups, the success rate did not differ signiifcantly between the two groups (93.75 % vs 93.85 %). Compared with transpancreatic septotomy with precutting techniques group, pancreatic duct guide wire group is less intubation time consuming (5.92 ± 0.69 vs 12.81 ± 3.67) min, the difference was statistically significant (t = -2.27,P < 0.05). 25 patients experienced complications, with 6 cases of acute pancreatitis, 2 cases of biliray tract infection in pancreatic duct guide wire group, and 8 cases of acute pancreatitis, 3 cases of hemorrhage,6 cases of biliray tract infection in transpancreatic septotomy with precutting techniques group. The pancreatic duct guide wire group had a signiifcantly lower incidence of complications (6.25 % vs 13.08 %). the difference was statistically signiifcant (χ2 = 3.27,P < 0.05). The incidence of acute pancreatitis did not differ signiifcantly between the two groups (4.69 % vs 6.15 %).ConclusionsPancreatic duct guide wire and transpancreatic septotomy with precutting techniques both can further improve the success rate of bile duct cannulation with ERCP. The incidence of acute pancreatitis did not differ significantly between two groups. But pancreatic duct guide wire group is less intubation time consuming, and had a significantly lower incidence of complications. Because of the convenience and safety of the pancreatic duct guide wire technique, and the insertion of the pancreatic duct does not increase the risk of postoperative acute pancreatitis. We think that this method is more worthy of Clinical promotion.

Objective To evaluate the role of opioid receptors and phosphatidylinositol 3-kinase/proteinserine-threonine kinases (PI3K/Akt) and extracellular signal-regulated kinase (ERK) signaling pathways in reduction of hypoxia/reoxygenation (H/R)-induced injury to cardiomyocytes by remifentanil preconditioning in rats.Methods Primary cardiomyocytes were obtained from adult male Sprague-Dawley rats and cultured in DMEM culture medium.The cells were seeded in 48-well plates (density 2 × 104 cells/ml,500 μl/well) and randomly divided into 12 groups (n =9 each):control group (group C),group H/R,hypoxia preconditioning group (group HPC),remifentanil preconditioning (RPC) group,naltrindole (δ receptor antagonist) + RPC group,nor-binaltorphimine (κ receptor antagonist) + RPC group (BNI + RPC group),wortmannin (PI3K inhibitor) + RPC group (W+ RPC group),PD98059 (ERK inhibitor) + RPC group (PD + RPC group),NTD group,BNI group,W group and PD group.In group H/R,the cardiomyocytes were exposed to 90 min of hypoxia,followed by 120 min of reoxygenation.In group HPC,the cardiomyocytes were exposed to 10 min of hypoxia,followed by 30 min of reoxygenation before H/R.In group RPC,the cardiomyocytes were preconditioned with remifentanil with the final concentration of 1 μmol/L for 10 min,followed by 30 min routine culture before H/R.In NTD + RPC,BNI + RPC,W + RPC and PD + RPC groups,naltrindole 5μmol/L (final concentration),nor-binaltorphimine 5 μmol/L (final concentration),wortmannin 0.1 μmol/L (final concentration) and PD98059 30μmol/L (final concentration)were added,respectively,and then the cells were coincubated with remifentanil for 10 min,followed by 30 min routine culture before H/R.The viability of cardiomyocytes,cell apoptosis and activity of lactate dehydrogenase (LDH) in the culture medium were detected.The apoptosis rate (AR) was calculated.Results Compared with group C,the viability of cardiomyocytes,AR and activity of LDH in the culture medium were significantly increased in group H/R (P ＜ 0.05).Compared with group H/R,the viability of cardiomyocytes,AR and activity of LDH in the culture medium were significantly decreased in HPC,RPC and BNI + RPC groups (P ＜ 0.05),and no significant changes were found in the parameters mentioned above in NTD + RPC,W + RPC,PD + RPC,NTD,BNI,W,and PD groups (P ＞ 0.05).The viability of cardiomyocytes was significantly lower,and the AR and activity of LDH in the culture medium were higher in NTD + RPC,BNI + RPC,W + RPC,and PD + RPC groups than in RPC group (P ＜ 0.05).Conclusion Remifentanil preconditioning activates PI3K/Akt and ERK signaling pathways possibly through activating δ opioid receptors thus attenuating H/R-induced injury to cardiomyocytes in rats.

Objective To evaluate the effects of morphine preconditioning on the expression of microRNAs (miRNAs) during hypoxia-reoxygenation (H/R) in isolated cardiomyocytes in rats with heart failure.Methods Healthy adult male Sprague Dawley rats,w eighing 200-220 g,were used in this study.Adriamycin 2.0 mg/kg was injected once a week for 6 weeks via the tail vein to induce heart failure.The cardiomyocytes were isolated from the failing hearts of rats and seeded in 24-well plates or in 60 mm diameter dishes.The cells were then randomly divided into 3 groups (n =16 each) using a random number table:control group (group C); group H/R;morphine preconditioning group (group MP).The cells were cultured in normal culture atmosphere in group C.After being exposed to hypoxic air (5％ CO2-95％ N2) for 90 min,the cells were returned to the high-glucose DMEM supplemented with 10％ newborn bovine serum and were then cultured for 120 min in H/R and MP groups.In group M,the cells were cultured in morphine culture medium (final concentration of morphine 0.3 μmol/L) for 10 min and then were returned to the culture medium without morphine and cultured for 30 min immediately before hypoxia.At 120 min of reoxygenation,the cells of 8 wells in each group were chosen to detect the cell viability and lactate dehydrogenase (LDH) activity (by Typan blue staining).All the RNAs were extracted from the cardiomyocytes of the left 8 wells in each group and subjected to miRNA microarray to screen differentially expressed miRNAs.Results The cell viability was significantly lower,the activity of LDH was higher,the expression of miR-6216 and let7e-5p was higher,and the expression of miR-133b-5p was lower in H/R and MP groups than in group C (P ＜ 0.05).Compared with H/R group,the cell viability was significantly increased,the activity of LDH was decreased,the expression of miR-133b-5p was up-regulated,and the expression of miR-6216 and let-7e-5p was down-regulated in MP group (P ＜ 0.05).Conclusion Morphine preconditioning reduces H/R injury to isolated cardiomyocytes in rats with heart failure through regulating the expression of miRNAs such as miR133b-5p,miR-6216 and let-7e-5p.

Objective To investigate the changes of hypoxia-inducible factor(HIF)-1α and hexokinase-Ⅱ(HK-Ⅱ)expression in human esophageal squamous cell carcinoma and its effect in glycolysis.Methods TE13 cells and Eca109 cells were cultured under hypoxic condition(1 ％O2)for different hypoxic time(6,12,24 and 48 hours).Cells cultured under normal oxygen condition(20％O2)were set as control.The changes of HIF-1α and HK-Ⅱ expressions at protein level were detected by Western blot.HIF-1α genes were specifically silenced with RNA interference technology(RNAi),and then the changes of HIF-1α and HK-Ⅱ expression were determined by realtime PCR and Western blot.Under normal oxygen and hypoxic condition,the changes of lactic acid concentration in cell culture medium were detected by spectrophotometric method.Results Under hypoxic condition,the expression of HIF-1α and HK-Ⅱ gradually increased as hypoxic time extended(P＜0.05),reached a peak at 12h and then gradually decreased as time extended.Compared with that under normal oxygen condition,the expression of HK-Ⅱ in TE13 cells and Eca109 cells significantly increased under hypoxic condition(P＜0.05),which was more significant after 12 hours hypoxia.The result of realtime PCR indicated that under normal oxygen condition the expression of HIF-1α at RNA level in TE13/shRNA cells and Eca109/shRNA cells significantly decreased compared with TE13 cells and Eca109 cells without interference(P＜0.05).The expression of HK-Ⅱ at RNA level was consistent with the result of HIF-1α.Under normal and hypoxia condition,the expression of HK-Ⅱ at protein level in TE13/shRNA cells and Eca109/shRNA cells significantly decreased compared with TE13 cells and Eca109 cells without interference,and the difference was statistic significant(P＜0.05).The lactic acid secretion of TE13 cells and Eca109 cells under hypoxia condition(14.707 ± 3.594 and 15.062 ±3.901)was higher than that under normal oxygen condition(6.070±1.839 and 6.891±1.592,P＜0.05).The lactic acid secretion of TE13/shRNA cells and Eca109/shRNA cells significantly decreased compared with TE13 cells and Eca109 cells without interference,and the difference was statistic significant(P＜0.05).Conclusion The expressions of HIF-1α and HK-Ⅱ in human esophageal squamous cell carcinoma significantly increased under hypoxia conditions.The expression of HK-Ⅱ is closely correlated with lactic acid concentration and HIF-1α expression.HIF-1α may affect cell glycolysis through HK-Ⅱ.

Objective To investigate the coagulation disorder status of patients with pancreatic cancer and to explore its role in tumor formation,progression and metastasis.Method The present study involved 114 patients with pancreatic cancer diagnosed by surgery or biopsy,and 40 healthy volunteers.For each individual,nine plasma coagulation parameters were tested using the STAGO Compact automated.Results The levels of plasma antithrombin Ⅲ (AT-Ⅲ) and protein C of the pancreatic cancer group were significantly reduced compared with the control group,while the levels of plasma PT,APTT,INR,FIB,F-Ⅷ,D-dimer (D-D) were significantly elevated.The level of plasma D-D increased with increase in clinical stage,while the level of AT-Ⅲ decreased.The level of plasma D-D became higher with worsening in histological grade.Conclusions Patients with pancreatic cancer were in a state of hypercoagulation,with reduced anticoagulation function and secondary hyperfibrinolysis.The level of Plasma D-D was significantly associated with the clinical stage,histological grade and distant metastasis.These together with AT-Ⅲ could be used as indicators to monitor patients with pancreatic cancer.

Objective To study the clinicopathological characteristics and the prognostic factors in metastatic pancreatic tumor.Methods The clinical data of 32 patients with metastatic pancreatic tumors were retrospectively analyzed.The survival of the patients were evaluated with the KaplanMeier method.Univariate analysis was done by log-rank test.Results The primary foei of these32 patients included:9 lung cancer,6 renal cell carcinoma,5 gastric cancer,12 others.The median survival was 15.5 months in the radical surgery group (n 8),18 months in the cryosurgery group(n=3),8 months in the group with palliative bypass operation (n=9),6 months in the group with local radiotherapy (n=3),and 5 months in the group without treatment (n=9).Univariable analysis revealed the type of primary tumor,treatment of the primary tumor,interval between diagnosis of primary tumor and pancreatic metastases,localized pancreatic metastasis,and treatment of the metastatic pancreatic tumor were associated with prognosis.Conclusions Metastatic pancreatic tumors are rare.Long term survival can be achieved with pancreatic resection in a highly selected subset of patients according to the type of primary tumor.Therapeutic cryosurgery as a new minimally invasive,targeted and immunologic method needs to be further evaluated.

Objective To study the clinicopathological characteristics,diagnosis and treatment of patients with mucinous cystic neoplasms (MCNs) of the pancreas.Methods The clinical data of 42 patients with mucinous cystic neoplasms of pancreas were retrospectively analyzed.Chi-square test and independent sample t-test were used for statistical analysis.Results The mean age of the patients was 53.1 yeas (ranged from 29 to 78 y).There were 32 female (76.2％).The patients were divided into two groups according to symptoms (the symptomatic group and the asymptomatic group).There were significant differences in tumor size,tumor location,operation type,operation time and adhesions with the surrounding tissues between the 2 groups of patients.The patients were then divided into three groups according to pathological type.There were significant differences in age,tumor marker,tumor location,operation type,operation time,adhesions with the surrounding tissues,operative blood loss,and postoperative hospital stay among the 3 groups of patients.There was no tumour recurrence in the patients with cystadenoma.Patients with cystadenocarcinoma developed tumor recurrence at a mean of 16.4 months (range,0-50) and died of tumor recurrence even after radical surgery at a mean of 22.9 months (range,3-58).Conclusion MCNs were seen most commonly in middleaged women.The presence of symptoms was a sign of malignancy.Resection of the tumor in the early stage is the key in dealing with MCNs.

Objective To analyse the diagnosis and treatment options of serous cystadenoma of the pancreas.Method The clinical data of 57 patients operated in the Tianjin Medical University Cancer Institute & Hospital from August 1996 to December 2011 with pathologically confirmed serous cystadenoma of pancreas after the operation were retrospectively studied.Results There were 13 males (22.8％) and 44 females (77.2％).The median age was 56.8 years.The patients were asymptomatic in 31.6％.CT was accurate in the diagnosis in 70.6％.All patients received surgical resection,inluding pancreaticoduodenectomy (n =17,29.8％),distal pancreatectomy (n =38,66.7％),palliative resection (n=1),and tumor enucleation (n=1).Postoperative complications developed in 6 patients.Histopathologically,there were 50 cases of serous microcystic adenoma (87.7％) and 7 cases of serous oligocystic adenoma (12.3 ％).One of these patients had developed into serous cystadenocarcinoma.At a follow-up of 12 months to 15 years,one patient with serous cystadenocarcinoma died 13 months after the operation.The remaining patients were all alive.Statistical analysis was performed based on the postoperative histopathological type and tumor size.The mean postoperative hospital stay of the group of patients with serous microcystic adenoma were significantly longer than the patients with serous oligocystic adenoma [(17.39±7.61) d vs (19.43±0.98) d,P=0.002].The incidence of patients with clinical symptoms was higher in the group of patients with tumor size ≥4 cm when compared with the patients with tumour size ＜4 cm.There was no significant difference on the other parameters.Conclusions Pancreatic serous cystadenoma is a rare pancreatic tumor,and it often happens in elderly women.Indications for surgical resection included symptomatic tumours,tumor diameter more than 4 cm,malignant biological behavior,malignancy could not be ruled out,and potentially malignant tumors.For asymptomatic patients and tumor size less than 4 cm,surgical resection should also be considered if the tumour progresses on follow-up.

Objective To evaluate the roles of 1-phosphatidylinositol 3-kinase (PI3K) and extracellular signal-regulated kinase (ERK) signaling pathways in reduction of ischemia-reperfusion (I/R) injury to the isolated hearts by morphine preconditioning in the rats with chronic heart failure.Methods Adult male Sprague-Dawley rats,weighing 200-230 g,in which doxorubicin 2.0 mg/kg was injected via the tail vein once a week for 6 weeks to induce chronic heart failure,were studied.At the end of 8th week,42 rats with chronic heart failure were randomly divided into 7 groups (n =6 each) using a random number table:sham operation group (group S),I/R group,morphine preconditioning group (group MP),PD98059 (ERK inhibitor) + morphine preconditioning group (group PD + MP),wortmannin (PI3K inhibitor) + morphine preconditioning group (group WT + MP),PD98059 group (group PD) and wortmannin group (group WT).The hearts were quickly excised and passively perfused in a Langendorff apparatus and subjected to 30 min of occlusion of the left coronary artery followed by 2 h of reperfusion to establish the model of I/R injury.In group S,the hearts were only sutured,but not ligated and were continuously perfused with K-H solution for 195 min.In group I/R,the hearts were perfused with K-H solution for 45 min before ischemia.In group MP,the hearts were perfused with K-H solution for 15 min,with K-H solution containing morphine 1 μmol/L for 5 min and then with K-H solution for 5 min (3 cycles in total) before ischemia.In PD + MP and WT + MP groups,the hearts were perfused with K-H solution containing PD98059 (10 μmol/L) and wortmannin (100 nmol/L),respectively,starting from 10 min before morphine preconditioning until 5 min of ischemia.In PD and WT groups,the hearts were perfused with K-H solution containing PD98059 (10 μmol/L) and wortmannin (100 nmol/L),respectively,starting from 40 min before ischemia until 5 rin of ischemia.At 15 min of equilibration (baseline) and 5 and 10 min of reperfusion,the coronary flow was collected to detect the activity of lactate dehydrogenase (LDH).Infarct size (IS) and area at risk (AAR) were measured at the end of reperfusion and IS/AAR ratio was calculated.Results Compared with group S,LDH activity was significantly increased at 5 and 10 min of reperfusion,IS and IS/AAR ratio were also increased (P ＜ 0.05),and no significant change was found in AAR in group I/R (P ＞ 0.05).Compared with group I/R,LDH activity was significantly decreased at 5 min of reperfusion,IS and IS/AAR ratio were also decreased (P ＜ 0.05),and no significant change was found in AAR in group MP,and no significant change was found in LDH activity,IS,AAR and IS/AAR ratio in WT and PD groups (P ＞ 0.05).Compared with group MP,LDH activity was significantly increased at 5 and 10 min of reperfusion (P ＜ 0.05),and IS and IS/AAR ratio were decreased in group PD + MP,and no significant change was found in LDH activity,IS,AAR and IS/AAR ratio in group WT + MP (P ＞ 0.05).Conclusion Activation of ERK signaling pathway is involved in reduction of I/R injury to isolated hearts by morphine preconditioning in rats with chronic heart failure,however,PI3K signaling pathway has no such effect.

Objective:To search for a method for radical resection of pancreatic and duodenal malignancy involving the mesentery root and for the long post-operation survival of patients.Methods:From Jan.2004 to Aug.2006,a total of 26(16 male and 10 female. aged 27-70)patients with pancreatic and duodenal malignancy involving the mesentery root were treated in our department.The patients included 3 with duodenal malignancy and 23 with pancreatic malignancy.Curative resection was performed by the extended pancreaticoduodenetomy(Whipple procedure)combined with mesentery root resection(MRR)for all patients.The outcomes,safety and the post-operation survival rate were analyzed retrospectively.Results:Thirteen patients were treated with Whipple procedures combined with MRR,9 were treated with partial portal vein/superior mesenteric vein(PV/SMV)and reconstruction of the vessel,and 4 patients received pre-shunt between PV and SMV with artificial vessel graft before the extended Whipple and MRR procedures.The operation time was 2.5 to 7(4.4?1.1)hour,and blood loss was 300 to 5 000(1892?1414)ml with the blood transfusion of 0 to 5 600(2 100?1 586)ml.There was no death in our group and 7(27%)had post-operation complication.The post-operation hospital stay was 10 to 30 days.The pathologic examination showed negative surgical margins for all specimens.The tumor size was 4 to 10 (6.17?2.03)cm.After a follow-up of 9 to 38 months,the pain was relieved in all patients.One of the 3 patients with duodenal adenocarcinoma had liver metastasis at 10 months after operation,and the other 2 survived 10 months and 27 months without evidence of tumor reccurence.The patient with pancreatic micro-adenocarcinoma died of local reccurence 9 months after operation.The patient with neuroendocrine carcinoma died of organ failure 24 months after operation.The patient with lymphoma have survived for 24 months after operation.The 1-year and 2-year accumulated survival rates in the 20 cases with pancreatic ductal cancer were 86.6% and 45.6%. respectively.Conclusion:The extended Whipple procedure with MRR is safe and effective.It can obtain R0 resection in patients with malignant tumors(over 5 cm in diameter)in the head,neck and uncinate process of the pancreas and duodenal.