Objective To investigate the mechanism by which pancreatic cancer cells(PCC) acquire drug resistance against 5-FU and gemcitabine. Methods The cytotoxic effects of 5-FU and gemcitabine on PCC ( Panc-1, Mia-Paca-2 andCapan-1) were assessed by using Sulforhodamine B and the expression of anti-apoptotic genes of the Bcl-x L and mcl-1 were analyzed by RNase protection assay and Western blot. Results5-FU and gemcitabine effect cytotoxicity towards PCC. After repeated treatment with 5-FU, the IC 50 values in Capan-1 cells increased by 2.1 fold (P

Objective To investigate the effect of dexmedetomidine on the expression of hypoxia-inducible factor-1α (HIF-1α) during hypoxia/reoxygenation (H/R) in human renal tubular epithelial cells.Methods Human renal tubular epithelial cells (HK-2 cells) cultured in vitro were randomly divided into 4 groups (n =24 each) using a random number table:control group (group C),dexmedetomidine group (group DEX),H/R group and H/R+ dexmedetomidine group (group H/R + DEX).In group C,the cells were incubated for 28 h in an incubator filled with normoxia at 37 ℃.In group DEX,dexmedetomidine 0.1 nmol/L (final concentration) was added to the culture medium and the cells were incubated for 2 h,and then incubated for 28 h in an incubator filled with normoxia at 37 ℃.In group H/R,the cells were incubated in an anaerobic chamber for 24 h at 37 ℃,and then incubated for 4 h in an incubator filled with normoxia at 37 ℃.In group H/R + DEX,the cells were incubated for 2 h in the culture medium containing dexmedetomidine 0.1 nmol/L (final concentration),incubated in an anaerobic chamber for 24 h at 37 ℃,and then incubated for 4 h in an incubator filled with normoxia at 37 ℃.After treatment in each group,the cell viability was measured by MTT assay,cell apoptosis was measured using flow cytometry,the expression of HIF-1α mRNA was detected using RT-PCR,the expression of HIF-1α and activated caspase-3 protein was detected by Western blot,and the cell growth was observed.The apoptosis rate was calculated.Results Compared with group C,the cell viability was significantly decreased,the apoptosis rate was increased,and the expression of HIF-1α mRNA and protein and activated caspase-3 protein was up-regulated in H/.R and H/R + DEX groups,and no significant change was found in group DEX.Compared with group H/R,the cell viability was significantly increased,the apoptosis rate was decreased,the expression of HIF-1α mRNA and protein was up-regulated,the expression of activated caspase-3 protein was down-regulated,and the cell status was significantly improved in group H/R + DEX.Conclusion The mechanism by which dexmedetomidine attenuates H/ R-induced damage to human renal tubular epithelial cells may be related to up-regulated expression of HIF-1 α and inhibited cell apoptosis.

Objective To investigate the effect of STH-2 cardioplegic solution containing levosimendan on ischemia-reperfusion (I/R) injury in isolated rat hearts. Methods Thirty-two male Wistar rats weighing 250-300 gwere anesthetized with intraperitoneal 3% pentobarbital 30 mg/kg. The hearts were rapidly excised and perfused with oxygenated (95% O2-5% CO2) K-H solution for 30 min in a Langendorff apparatus and then divided into 4groups (n = 8 each) according to the composition of cardioplegic solution: group Ⅰ control (group C) was perfused with STH-2 cardioplegic solution; group Ⅱ , Ⅲ and Ⅳ were peffused with STH-2 cardioplegic solution containing levosimendan 0.03 μmol/L (L1), 0.3 μmol/L (L2) and levosimendan 0.3 μmol/L + glibenclamide 10 μmol/L (L2+ G) respectively. The isolated hearts were first perfused with different cardioplegic solutions for 2 h and then with K-H solution for 30 min. The coronary effluent was collected before ischemia (baseline) and at 10, 20 and 30 min of reperfusion for measurement of creatine kinase (CK) and lactate dehydrogenase (LDH)activities. Myocardial specimens were obtained from apex at 30 min of reperfusion for determination of myocardial ATP and MDA contents and SOD activity. Results Perfusion with STH-2 cardioplegic solution significantly increased CK and LDH activities and MDA content, and significantly decreased SOD activity. Levosinendan 0.03or 0.3 μmol/L significantly attenuated the cardioplegia-induced increase in LDH,CK and SOD activities and MDA content. The protective effects of levosimendan on myocardium against I/R injury were reversed by glibenclamide to some extent. Conclusion Levosimendan can protect myocardium from I/R injury in a dose-dependent manner by opening KATP channel.

Objective:To evaluate the role of nuclear factor erythroid 2-related factor/ heme oxygenase-1 (Nrf2/HO-1) signaling pathway in dexmedetomidine-induced reduction of oxygen-glucose deprivation and restoration (OGD/R) injury to microglia.Methods:BV-2 microglia were cultured in high-glucose DMEM culture medium supplemented with 10% fetal bovine serum in an normal culture incubator at 37 ℃ (5%CO 2-21%O 2-74 %N 2). The cells were seeded in 96-well plates at a density of 1.5×10 4 cells/ml (200 μl/well) or 6-well plates at a density of 2×10 5 cells/ml (2 ml/well) and divided into 5 groups ( n=30 each) using a random number table method: control group (group C), dexmedetomidine group (group D), group OGD/R, OGD/R+ dexmedetomidine group (group OGD/R+ D) and OGD/R+ dexmedetomidine+ ML385 group (group OGD/R+ D+ ML). The cells in group C were continuously cultured in a normal culture incubator for 26 h. In group D, dexmedetomidine at the final concentration of 10 μmol/L was added, cells were incubated for 2 h, and then were continuously incubated in a normal culture incubator for 26 h. In OGD/R, OGD/R+ D and OGD/R+ D+ ML groups, the culture medium was replaced with glucose-free DMEM culture medium, cells were cultured for 2 h in an incubator at 37 ℃ (5%CO 2-1%O 2-94 %N 2), the culture medium was replaced with high-glucose DMEM culture medium containing 10% fetal bovine serum and then the cells were cultured for 24 h in a normal incubator.Dexmedetomidine at the final concentration of 10 μmol/L was added at 2 h before OGD in OGD/R+ D and OGD/R+ D+ ML groups.Nrf-2 inhibitor ML385 at the final concentration of 4 μmol/L was added at 30 min before dexmedetomidine was added in group OGD/R+ D+ ML.Cells in 6 wells in each group were selected randomly for assessment of cell viability (by methyl thiazolyl tetrazolium assay) and apoptosis (using flow cytometry), and for determination of the concentrations of tumor necrosis factor (TNF)-α, interleukin (IL)-6, and IL-10 in the supernatant (using enzyme-linked immunosorbent assay), the expression of Nrf2 in nucleus, Nrf2 and HO-1(by Western blot ) and the expression of HO-1 mRNA (by real-time polymerase chain reaction). Results:Compared with group C, the cell viability was significantly decreased, cell apoptosis rate and concentrations of TNF-α, IL-6 and IL-10 in the supernatant were increased, and the expression of Nrf2 in nucleus, Nrf2, HO-1 and its mRNA was up-regulated in OGD/R and OGD/R+ D groups ( P<0.05), and no significant change was found in each parameter mentioned above in group D ( P>0.05). Compared with group OGD/R, the cell viability and IL-10 in the supernatant concentration were significantly increased, cell apoptosis rate and concentrations of TNF-α and IL-6 in the supernatant were decreased and the expression of Nrf2 in nucleus, Nrf2, HO-1 and its mRNA was up-regulated in group OGD/R+ D ( P<0.05), and no significant changes were found in the parameters mentioned above in group OGD/R+ D+ ML ( P>0.05). Compared with group OGD/R+ D, the cell viability and concentration of IL-10 in the supernatant were significantly decreased, cell apoptosis rate and concentrations of TNF-α and IL-6 in the supernatant were increased and the expression of Nrf2 in nucleus, Nrf2, HO-1 and its mRNA was down-regulated in group OGD/R+ D+ ML ( P<0.05). Conclusion:The mechanism by which dexmedetomidine alleviates OGD/R injury to microglia may be related to promoting the activation of Nrf2/HO-1 signaling pathway and inhibition of inflammatory responses.

Objective To investigate the expression of neurokinin 3 receptor (NK 3R) in normal pancreas and pancreatic cancer, and to study the localization of this receptor in pancreatic cancer Methods The expression of NK 3R mRNA was investigated using reverse transcription polymerase chain reaction (RT PCR) in normal pancreas and pancreatic cancer tissues The protein level of NK 3R was investigated by Western blot Immunohistochemistry was used to localize expression site of NK 3R Results NK 3R mRNA was overexpressed in pancreatic cancer tissue when compared with normal pancreas NK 3R protein was 1 1?0 5 in normal pancreas and 14 6?3 6 in pancreatic cancer, P

Objective To investigate the clinical value of multi-slice spiral CT in the evaluation of esophageal variceal bleeding .Methods 50 cirrhosis patients with esophageal varices received multi-slice spiral CT and gastroscopy detection .The application value of multi-slice CT in the assessment of esophageal bleeding was evaluated according to the results of gastroscopy detection .Results CT angiography score had significantly positive correlation with the severity of endoscopic varices and endoscopic red color sign (r=0.762,0.687,all P<0.01).The sensitivity and specificity of CT angiography score in diagnosis of endoscopic red signs RC 3 were 76.92% and 92.50%. Conclusion The results of multi-slice CT and gastroscopy are positively correlated with the severity of esophageal varices,which can be used to predict the risk of esophageal bleeding .

Objective To investigate the role of substance P (SP) and its neurokinin-1 receptor (NK-1R) in the pathophysiologic process of Crohn′s disease. Methods In 23 surgical patients of Crohn′s disease and 24 healthy controls, reverse transcription polymerase chain reaction (RT-PCR) was used to determine the mRNA expression of NK-1R, Western blot analysis was used to determine NK-1R protein expression levels, and immunohistochemistry was used to localize expression of NK-1R. Results Compared with normal gut NK-1R mRNA and NK-1R protein in Crohn′s disease were overexpressed. In Crohn′s disease moderate to strong intestinal NK-1R immunoreactivity was found in the lamina propria mononuclear cells, lymphoid follicles, and the surface and crypt epithelium, lymphoid follicles, submucosal vessels, smooth muscle and myoenteric plexus neurones. Conclusions In cases of Crohn′s disease, overexpression of NK-1R may disturb neuropeptides loop balance, and may be involved in the pathophysiological change in this disease.

AIM: To study the expression and the role of neurokinin-1 receptor(NK-1R) and neurokinin-2 receptor (NK-2R) in distal ileum during acute necrotizing pancreatitis(ANP).METHODS: 130 adult Sprague-Dawley rats were divided into 2 groups: the rats in ANP group( n=80 ) were induced by the retrograde intraductal infusion of 5% sodium taurocholate. The rats in sham operation control group ( n=50 ) received laparotomy only. Reverse transcription polymerase chain reaction (RT-PCR) was used to determine the mRNA expression of NK-1R and NK-2R, Western blot was used to investigate expression level of NK-1R.RESULTS: NK-1R and NK-2R mRNA levels were enhanced in the distal ileum of ANP rats compared with controls. Western blot revealed stronger NK-1R immunoreactivity exited in intestinal mucosa in ANP rats. The overexpression of NK-1R was associated with mucosal pathological score( r=0.77,P

AIM: To investigate the expression of neurok inin-2 receptor (NK-2R) in normal pancreas and chronic pancreatitis (CP) tissues. The relation of expres sion of NK-2R with pain in CP was also evaluated. METHODS: CP tis sues were ob tained from 18 men and 7 women undergoing pancreatic head resection as a result of CP. Normal human pancreatic tissues were obtained from 20 patients (11 males/ 9 females). Real-time quantitative reverse transcription polymerase chain reacti on was used to determine the mRNA expression of NK-2R, Western blot analysis was used to determine its protein expression level, and immunohistochemistry was us ed to localize expression site of NK-2R protein. NK-2R mRNA level and pain were also analysed whether correlation exists. RESULTS: NK-2R mRNA and p rotein level were enhanced expressed in CP tissue samples compared with normal pancreas. Ove rexpression of NK-2R was related to the intensity ( r=0 59, P

Objective To investigate the clinical characteristics and angiographic findings of cardiogenic shock(CS)following acute myocardial infarction(AMI) in elderly patients.Methods Between January 2015 and April 2016,we carried out a retrospective observational analysis of consecutive elderly patients in Tianjin Chest Hospital,who suffered CS-complicating AMI.Emergency angiography and percutaneous coronary intervention(PCI) were performed after admission.All selected patients were divided into CS and non-CS groups according to whether CS occurred.Electrocardiograph (ECG),cardiac enzyme testing,and ultrasound cardiography were performed after admission to monitor the occurrence of CS.Results The incidence of CS-complicating AMI was 8.33％ (34/408) in elderly patients.Among all CS patients enrolled,the aged patients accounted for 91.89 ％ (3 4/3 7).In-hospital mortality rate was 2 9.41 ％ (10/3 4).There were significant differences between two groups in WBC,H s-CRP,blood glucose,CR and ALT (t =2.403,4.596,6.778,6.109,each P＜0.05).The NT-Pro BNP level,the time of FMC,the frequency of left main and multivessel disease were higher in the CS group than in the non-CS group (each P ＜ 0.05).Conclusions Elderly patients are bearing high risk of CS following AMI.Prolonged FMC time and the presence of left main and/or multivessel lesion are independent risk factors for the development of CS.The optimal revascularisation strategy can improve the clinical outcome of patients with CS.