Objective This study was to evaluate the effects of growth hormone (GH) on cell apoptosis of intestinal epithelium in acute necrotizing pancreatitis (ANP) rats.Methods Seventy-two SD rats were divided into three groups randomly: sham operation (SO) group, ANP group, and GH treatment group. GH-treatment group received subcutaneously 0 75?U/kg of GH. At 3?h, 6?h, 12?h, and 24 ?h after induction of ANP, the small intestinal specimens were harvested and apoptosis of intestinal epithelium was studied by DNA gel electrophoresis, FCM and TUNEL method. FasL and Bax protein expressions were detected by immunohistochemistry.ResultsThe ladder pattern on DNA gel electrophoresis presented in all time points of ANP group, but only seen in GH-treated group at 3h. Apoptotic percentage significantly increased in ANP group [(50?11)%,(80?9)%,(48?17)%,(50?10)%] as compared with SO group [(54?4)%, (28?6)%, (39?5)%,(29?11)%], all P

Objective To investigate the deleterious effects of enterogenous endotoxemia on the course of acute pancreatitis in mice, and its possible mechanisms. Methods Sixty five C57BL/6 mice were assigned to 4 groups randomly, including acute edematous pancreatitis (AEP), lipopolysacchride (LPS), AEP plus LPS and normal control. AEP was induced by intraperitoneal injection of cerulein with a dosage of 50 ?g/kg at hourly interval for seven times under ether anesthesia. LPS was administrated via a gastric tube with a dosage of 5 mg/kg at 6 h after the first cerulein or saline injection. Serum amylase and LDH activities were measured at 12 h, 24 h, 48 h and 5 d. Pathological alteration in pancreas was studied. Expressions of Mac 1 (CD11b/CD18), P selectin, E selectin and ICAM 1 were evaluated by using inmmunohistochemical procedures. Expressions of cytokine genes were determined by means of RT PCR and Southern blot. Myeloperoxidase (MPO) activity in pancreas was also analyzed. Results Cerulein induced a typical changes of AEP in mice, which was confirmed by pathological changes and hyperamylasemia. LPS alone didn't develop either morphological changes or biochemical alterations. However, cerulein induced AEP challenged by LPS could cause marked parenchymal necrosis and hemorrhage with significant increment of serum amylase and LDH activities. Expressions of Mac 1, P selectin, E selectin and ICAM 1 in pancreas were enhanced. Cytokine genes including TNF?,IL 1?,IL 6 and IFN? mRNA were also upregulated. MPO activity was increased. Conclusion This study suggested that enterogenous endotoxemia, which could not induce pancreatic injury per se , could induce AEP into ANP in mice. Over stimulation of neutrophils and releasing of pro inflammatory mediators might be the contributing factors.

Objective To investigate whether apoptosis of intestinal epithelial cells occurs at the early stage of acute necrotizing pancreatitis (ANP) in rats. Methods Fourty eight Spraque Dawley rats were used. ANP model of rats was induced by retro injection of 5% sodium taurocholate into biliopancreatic duct. Laparotomized animals without induction of ANP (sham operation) served as controls. The distal segment of ileum specimens were harvested at 3 h, 6 h, 12 h and 24 h after operation and the apoptosis of intestinal epithelial cells was studied by DNA gel electrophoresis, FITC conjugated annexin V and propidium iodide (PI) staining cells analyzed by Flow Cytometry (FCM) and immunohistochemical procedures (TUNEL method). Results The DNA electrophoresis showed that typical “ladder” patterns appeared at all indicated time points in ANP group, while the DNA specimens from control group presented a single chromosomal lane, except the one at 12 h. Apoptotic percentage of detached intestinal epithelial cells assayed using Annexin V kit by FCM were (53.7?3.7)%, (27.6?6.0)%, (39.0?4.8)%, (29.0?11.3)% at 3 h, 6 h, 12 h and 24 h in control group and (50.3?11.3)%, (79.7?9.2)%, (47.8?17.3)%, (49.6?9.5)% in ANP group. There was a significant difference between two groups at 6 h, P

Objective Some experimental results indicated that cyclooxygenase 2 (COX 2) was involved in the regulatory process of balance between cell apoptosis and multiplication. The present study was designed to investigate the effect of COX 2 on the course of pancreatic adenocarcinoma cell apoptosis and its possible signal pathway. Methods Apoptosis of pancreatic cancer cells (PC 3, highly expressed COX 2) induced by selective COX 2 inhibitor, Celebrex (IC 50 , 100 ?mol), was detected by using DNA gel electrophoresis, flow cytometry and electron microscopy. Expressions of apoptotic related genes mRNA, including bcl xl, bax, Survivin, were analyzed by reverse transcription polymerase chain reaction (RT PCR). Results Substantial apoptosis was induced by the treating PC 3 cells with Celebrex, as revealed by typical ladder pattern of DNA fragments under DNA electrophoresis, increment of apoptotic apportion, and apoptotic body under electron microscopy. Apoptotic inhibitory genes, bcl xl, bax, Survivin, were expressed in PC 3 cells, and were down regulated significantly by Celebrex in bcl xl and Survivin but not for bax. Conclusion Above findings suggest that the COX 2 pathway contributes to the apoptosis of pancreatic cancer cell, which may be via signal transduction of bcl xl and Survivin genes.

Objective To investigate the influence of Lipoprotein lipase (LPL)/Hepatic lipase (HL) on hyperlipidemie acute necrotizing pancreatitis (HLANP) in rats. Methods The rats were fed with hyperlipidemic feed for 4 weeks, then the rats were injected with 5% sodium taurocholate into pancreatic duct to induce HLANP model. Seventy-two rats were randomly assigned to HLANP and control groups, and then each group was subdivided into 6 subgroups (n = 6) at 0, 3, 6, 12, 24 and 48 h. Serum amylase, cholesterol, triglyeride (TG), free fatty acid (FFA) levels and serum LPL, HI. activities were determined. Under the light-microscopy and electron microscopy, the histopathologic and uhrastructure changes of pancreas were observed; the HL mRNA expressions were detected by RT-PCR; HL protein expressions HL were assessed by immunohistoehemical staining. Results The serum amylase levels reached peak values at 12 h after ANP induction in the two groups, the mean values were 7 176U/L and 6 366U/L, which were significantly higher than those of baseline values (P <0.05) ; serum levels of cholesterol in HLANP group at 0 ～ 12 h were higher than those of control group, however, only at 0 h the difference (1.19±0.49 vs 0. 32±0.14 mmol/L) was significant (P < 0.05) ; serum levels of FFA in HLANP group were not significantly different when compared with those of control group; serum levels of LPL and HL at 3 h were (17.5±7) U/L and (18.6±3.9) U/L, which were significantly higher than those of control group (8.9±3.4 U/L and 9.5±2. 1 U/L, P < 0.05). The pancreatic tissues necrosis levels were significantly increased in HLANP groups (3, 6, 24 and 48 h) than those of control group at corresponding time points (P < 0.05). lipid droplet deposition, rough endoplasmic reticulum distension, zymogen granule reduction, and chondriosome swelling in acinar cells of pancreatic tissues in HLANP group were found. The HI, mRNA expressions at 3 h and 6 h in HLANP group were 1.1±0.09 and 0.89±0.08, which were significantly higher than those in control group (0. 11 ± 0.01 and 0.15±0. 03, P <0.05). HL proteins were positively expressed in pancreatic tissues of two groups before ANP was induced, and HL proteins were strongly positively expressed after ANP induction. Conclusions Lipase (LPL/HL) expression increased in HLANP rats, and the content of serum protein increased, which resulted in lipids decomposition and increased the severity of ANP. LPL/HL may be one of the key lipids metabolic enzymes aggravating HLANP.

Objective To observe the activation of pancreatic stellate cells (PSC) during the formation of pancreatic fibrosis induced by the pancreatic injection of trinitrobenzene sulfonic acid (TNBS). Meanwhile, the effects of PSC-related factors, such as transforming growth factor ? 1 (TGF-? 1), collagen Ⅰ and MMP-2 on the pathogenesis of pancreatic fibrosis in rats were also evaluated. Methods Pancreatic fibrosis model in rats was induced by the injection of 2% TNBS in ethanolate-phosphate buffer solution into the pancreatic duct. The rats were sacrificed and the pancreata were removed at the 72nd hour, 3rd week, 4th week, 5th week, 6th week and 7th week after the operation respectively. Expressions of ?-smooth muscle actin (?-SMA), transforming growth factor ? 1 (TGF-? 1), collagen Ⅰ and MMP-2 were determined by either immunohistochemistry or RT-PCR, or Western blot respectively. The ultrastructure of pancreas was studied by electron microscope at different time points. Results The inflammation, swelling and necrosis were the major pathological changes of the pancreas at the early stage after the injection of 2% TNBS. Subsequently, the fibrotic manifestations such as proliferation of the fibrosis, atrophy of vesicles, deposition of collagen because prominent at the 3rd week after the operation, which peaked at 4th week. The expression of TGF-? 1 was increased significantly at the 3rd week after the operation and reached maximum at the 4th week. The expression of ?-SMA, which indicated the activation of PSC, could be detected at the 3rd week and also reached the peak value at the 4th week. After wards, it was decreased gradually. During the first 72 hours, the expression of MMP-2 mRNA was increased significantly and then was fluctuated but still higher than that in normal rats. The deposition of type Ⅰ collagen was increased in the areas of fibrotic tissues. Conclusions PSC might involve in the courses of the development and progression of TNBS induced pancreatic fibrosis in rats. This action was achieved via the activation of PSC by TGF-? 1, the production of those extracellular matrix metabolic associated enzymes such as the synthesis of collagen Ⅰ and the secretion of MMP-2.

Objective To investigate the effects of cyclooxygenase-2 (COX-2) on the expressions of vascular endothelial growth factor (VEGF) and prostaglandin E_2 (PGE_2) in pancreatic carcinoma both in vitro and in vivo, and to clarify the possible mechanism of PGE_2 in mediating COX-2 inducing angiogenesis of pancreatic carcinoma. Methods In vitro study, the inhibitory effects of Celebrex, a selective cyclooxygenase-2 inhibitor, on the expression of VEGF and PGE_2 in pancreatic carcinoma cell lines PC-3 were determined using either enzyme-linked immuno-absorbent assay (ELISA) or radioimmunoassay (RIA). Effect of exogenous PGE_2 on the down-regulation of VEGF by Celebrex was also assessed. In vivo study, PC-3 cell line xenograft nude mice model was established. Changes of VEGF expression and PGE_2 of tumor tissues after the treatment of Celebrex were investigated using Western blotting or RIA. Results Celebrex suppressed the expressions of VEGF and PGE_2 in cultured PC-3 cell line with a manner of dose- and time-dependence. Exogenous PGE_2 up-regulated the expression of VEGF, which was suppressed by Celebrex in a dose-dependent fashion. In vivo study, administration of Celebrex into xenograft nude mice inhibited expressions of VEGF and PGE_2 significantly. Conclusion COX-2 is involved in angiogenesis in pancreatic carcinoma probably through the inhibition of the production of angiogenic factors such as VEGF. PGE_2 is likely to act as an important mediator in this process.

Objective To evaluate the protective effect by inhibiting nuclear factor-kappa B (NF-?B) by N-acetyl-L-cysteine(NAC) on donor rat lung. Methods 24 SD rats were randomly divided into two groups, the control group and the experiment group. The harvested lung blocks were flushed with and stored in the low-potassium-dextran (LPD) solution in control group, but in the experiment group, LPD solution containing NAC was used. The lungs were preserved at 4℃ for 16 hours in both groups. Isolated rat lung reperfusion models were established and the donor lungs were perfused for 1 hour. PaO_2 and PAwP were measured at every 15 min intervals during reperfusion. After reperfusion, the lung tissue wet-to-dry (W/D) ratio and myeloperoxidase(MPO) activity were obtained. The protein and mRNA expressions of intercellular adhesion molecule-1(ICAM-1) and nuclear factor-kappa B(NF-?B) were also observed by using immunohistochemistry and semi-quantitative RT-PCR at the end of reperfusion. Results After reperfusion for 60 minutes, in experiment group, PaO_2 drop in and increase PAwP lower than in control group (P

Objective To investigate lipopolysaccharide (LPS) tolerance and its possible mechanism in experimental acute pancreatitis(AP) mice. Methods Two hundreds and ten C56BL/6J mice were randomized into normal saline(NS)+LPS group( n =105)and AP +LPS group( n =105). Both groups were subdivided into seven groups according to different dose of LPS. AP model was induced by intra- abdominal administration of cerulein (50 ?g/kg) for seven times at 1 hour interval. LPS was given 6 hours after first cerulein injection . Cerulein was replaced by NS in NS+LPS group. Ten mice in each sub- group were randomly selected to investigate mortality rate for 7 days. Another 5 mice were killed at 12 hours after the first cerulein injection. Liver, lung, kidney, pancreas and serum were reserved to evaluate pathological changes and measurement of amylase (AMS) and lactate dehydrogenase (LDH) levels. Gene expression profiles of leucocyte in NS+LPS(15 mg/kg)subgroup and AP+LPS(15 mg/kg)subgroup were studied with oligonucleotide microarrays of 12 479 full length mouse genes respectively for three times to screen the different genes between two groups. Results Mortality rates in both groups were increased, and correlated with the dosage of LPS. Mortality rate in AP+LPS group was significantly lower than that in NS+LPS group with the same LPS dose( P

Hospital TQM system is designed to fit China′s specifics,based on the Total Quality Management (TQM)theory and integrating the system theory concepts, comprising two structural modules of perceived quality and non-perceived quality.The former is a trinity framework made up of patient satisfaction,employee commitment and social reputation,while the latter is a management system featuring“Six pillars and six columns”,constituting a three-dimensional management for integrated operation and integration of the hospital.