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 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

The etiology and pathogenesis of Crohn’s disease(CD)are not fully clear,and genetic susceptibility, immunologic disorder,intestinal barrier dysfunction and intestinal microecology are considered to be involved in the pathogenic mechanism of CD. In recent years,the relationship between intestinal microecology and CD has received much attention. Several studies confirmed that the intestinal microecology in CD patients was different from that in normal person. The change of intestinal microecology was correlated with the occurrence of CD,and modulation of intestinal flora was effective in the treatment of CD. This article reviewed the relationship between intestinal microecology and CD and the therapeutic prospect of intestinal microecology for the treatment of CD.

Objective To observe the protective effect of dexamethasone, the inhibitor of nuclear factor-Kappa B, on the ischemia-perfusion injury of rat lung during the period of lung preservation. Methods Twenty-four rats were randomly divided into two groups: the control group and the trial group. The harvested lung blocks were flushed with and stored in the low-potassium-dextran (LPD) solution in control group, but in the trail group LPD containing dexamethasone solution was used. The lungs were stored at 4 ℃ for 16 h in both groups. The isolated rat lung reperfusion models were established and the donor lungs were perfused for 1 h. 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 expression of intercellular adhesion molecule-1 (ICAM-1), nuclear factor-Kappa B (NF-?B) was also detected by using immunohistochemistry and semi-quantitative RT-PCR at the end of reperfusion. Results The levels of decreased PaO_2 and increased PawP in trail group were lower than in control group at the every interval time in the samples obtained 15 min after reperfusion (P

The effects of tetramethylpy-razine (TMP) on pancreatic blood flow and survival rate were studied in sodium tarocholate-in-duced acute pancreatitis (AP) in rats. The results showed that pancreatic relative blood flow and pancreatic tissue perfusion were significantly increased and the pathologic changes and survival rate were improved in TMP treated group-s. Plasma value of TXB2?6-Keto-PGF1? and platelet aggregation rate (PAR) were also mea-sured. We found that TMP could maintain the balance between TXA2 and PGI2 and lower the elevated PAR. It was suggested that TMP has therapeutic effect on AP in rats through improving pancreatic microcirculation; which was related to the maintanance of the balance between PGI2 and TXA2.

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