ObjectiveTo investigate the relationship between the expression of triggering receptor1 present on myeloid cells ( TREM-1 ) in intestinal tissue and intestinal barrier dysfunction in severe acute pancreatitis (SAP). MethodsSixty-four male Wistar rats were randomly (random number) divided into sham operation group ( SO group, n = 32) and SAP group ( n = 32 ). The SAP model was established by retrograde injection of 5％ sodium deoxycholate into bile-pancreatic duct. Specimens from blood and intestinal tissue were collected 2, 6, 12 and 48 hours after modeling. The levels of D-lactate, diamine oxidase (DAO) and endotoxin in serum were measured with an modified spectro-photometric method. The expressions of TREM-1, IL-1β and TNF-αt mRNA in terminal ileum were detected by RT-PCR. All data were processed with SPSS version 16. 0 package to make one-way ANOVA and Spearman correlation analysis. ResultsThe serum levels of D-lactate, DAO and endotoxin were significantly increased at all intervals in SAP group compared with SO group ( P ＜ 0. 05 ). The expressions of TREM-1, IL-1β and TNF-α mRNA in terminal ileum of rats in SAP group at all intervals were significantly higher than those in SO group (P ＜ 0. 05 ). The expression of TREM-1 mRNA was positively correlated with expressions of IL-1 β and TNF-α mRNA ( r = 0. 956, P = 0. 044; r = 0. 986, P = 0. 015 ), but correlation was not found between expressions of IL-1β mRNA and TNF-α mRNA ( P = 0. 133 ). ConclusionsThe expression of TREM-1mRNA in intestinal tissue of rats with SAP is elevated, leading to the release of inflammatory cytokines and intestinal mucosal injury, indicating TREM-1 might play an important role in the genesis of intestinal barrier dysfunction in rats with SAP.

Objective To explore the preparation of liposomal clodronate and investigate its inducing effects on the apoptosis of peritoneal macrophages in rats after severe acute pancreatitis (SAP).Methods Liposomal clodronate was prepared by means of thin film. SAP rat model was established by retrograde injection of 5% sodium taurocholate into the pancreatic duct. The peritoneal macrophages were obtained from SAP rats. After exposure to different doses of liposomal clodronate (50, 100,150 μl), the PM proliferation was determined by MTT colourimetry. The apoptosis of PM was measured by flow cytometry and agarose gel electrophoresis, respectively. Results The prepared liposomal clodronate had a suitable encapsulation efficiency of clodronate (5.8%) with an average size of 200 nm. The spherical shape of liposome was confirmed by transmission electron microscope. Exposed to liposomal clodronate of different doses resulted in a obvious growth depression (P＜0.01). The apoptotic rate of the PM was (10.32±0.34) %, (18.16±0.49)% and (29.87±0.35)% in three different dose groups and the difference was marked (P＜0.01). 1.2% of agarose gel electrophoresis of DNA extracted from apoptotic macrophages induced by liposomal clodronate showed clearer and characteristic ladder following the liposomal clodronate concentration. Conclusion Liposomal clodronate has a definite effect on peritoneal macrophages in SAP rats.

Objective To study the effects of elodronate-liposome on inducing apoptosis of alve-olar macrophages from rats with acute neetotizing pancreatitis (ANP).Methods The AMs of eight rats with ANP were isolated, purified then incubated from broehoalveolar lavage by the differing rates of attachment of the various cell types in a forty-well cell culture plate.Then they were randomized in-to five groups including control group,blank liposome group( 50 μ1, 100 μ1),clodronate-liposome group (50μ1,100μ1).Values of OD were determined by MTT.AO fluorescence and haematoxylin dye were employed to determine the apoptosis of the AMs.Results There were no significant differences be-tween control group and blank liposome group(50 μ1, 100 μ1).Significant differences were found be-tween control group and clodronate-liposome group(50 μ1, 100 μ1).There were no marked differences between blank liposome group(50μ1, 100 μ1)and clodronate-liposome group(50 μ1,100μ1).AO fluo-rescence and haematoxylin dye were available to define the apoptosis of the AMs.Conclusion Clodr-onate-liposome can effectively induce the apoptosis of the AMs.

Objective To investigate the apoptosis of Kupffer cell (KC) induced by lipsomal clodronate in rat with acute necrotizing pancreatitis (ANP). Methods Lipsomal clodronate was prepared by means of thin film, the model of ANP was established by injection of 5% sodium taurocholate of 4 ml/kg into the pancreatic capsule. The Kupffer cells were obtained from ANP rat. After exposure to different doses of lipsomal clodronate (0, 50, 100, 150 μl) , then the proliferation and apoptosis of KC was measured by MTT, flow cytometry and agarose gel electrophoresis of DNA. Results The prepared lipsomal clodronate had an average size of 100～200 nm, the spherical shape of liposome was uniform and confirmed by transmission electron microscope. When exposed to different concentration of lipsomal clodronate for 24 h, the growth suppression rate was 17. 4% , 24. 2% and 31. 1% , respectively, while the apoptosis rate of the KC was (14. 12 ±0.37)% , (18.74±0.43)% and (27.51 ±0.39)%, respectively; the difference was statistically significantly (P<0. 01) , the DNA of KC began degradation and gradually showed clear and characteristic ladder. Conclusions Lipsomal clodronate could induce apoptosis and suppress the growth of Kupffer cells in ANP rats.

Objective To investigate the bcl?2 gene modification on neurological function recovery in rats with spinal cord injury in neural stem cell transplantation. Methods Cultured rat neural stem cells by Ad?EGFP as vector?mediated side B?cell lymphoma 2 gene ( bcl?2 ) gene transfection of neural stem cells were divided into 3 groups: control group, negative transfection group, bcl?2 transfection group. Use western?blot to detect the expression of bcl?2 protein in neural stem cells before and after transfection. 85 adult female SD rats, successful model 72, were randomly divided into control group, NSCs group, bcl?2?NSCs groups, 24/group, rat acute spinal cord injury model in accordance with a modified Allen’ s method. Assess the motor function by BBB rating and the swash plate test. 7 days after modeling by RT?PCR and Western blot detection of spinal cord injury around HSP27, c?fos gene expression, TUNEL assay apoptosis. Four weeks after model drawn line HE staining and fluorescence microscopy EGFP?labeled NSC survival and distribution of the rats neurophysiological recovery by SEP and MEP. Results bcl?2 gene transfection of rat neural stem cells, bcl?2 transfection group and control group, negative transfection group compared to bcl?2 mRNA and protein levels were expressed ( P < 0. 05 ); lower extremity motor function in rats evaluation of bcl?2?NSCs group than NSCs group, NSCs group than the control group. 72 hours after modeling, bcl?2?NSCs number of apoptotic cells were significantly lower than the control group and NSCs group (P < 0. 05). 7 days after modeling, compared with the control group and NSCs group, bcl?2?NSCs group HSP27 gene and protein expression was significantly higher than that (P < 0. 05), bcl?2?NSCs group c?fos mRNA and protein expression was significantly reduced compared (P < 0. 05). 4 weeks after modeling, HE staining control group showed spinal cord tissue loss and the formation of syringomyelia, no axonal through. NSCs group damage zone few of neuraxis?like structures, syringomyelia smaller, bcl?2?NSCs group showed more nerve axon?like structure, no syringomyelia. EGFP?positive cells labeled:bcl?2?NSCs group the most, NSCs group followed, no control group, and the difference between the groups was statistically significant (P < 0. 05). After the 4th week, SEP and MEP latency period:bcl?2?NSCs group NSCs group > control group, and between the groups was significant difference ( P < 0. 05 ) . Conclusions By Ad?EGFP as vector?mediated side B?cell lymphoma 2 gene (bcl?2) gene transfection make neural stem cells can promote cultured rat neural stem cells. bcl?2 gene?modified neural stem cell transplantation can promote the regeneration of spinal cord injury synaptic elevated HSP27 expression after spinal cord injury, reduced expression and neural cell apoptosis after spinal cord injury bcl?2 gene and improve limb movement in rats function and electrophysiological function.