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.

Pancreatic encephalopathy(PE)is one of the severe complications in acute pancreati-tis,which is characterized by a group of neurological signs and symptoms. Itˊs very difficult to have an early diagnosis,while the morality of PE is very high and the prognosis is very poor. Eliminating causes of pan-creatic encephalopathy,early diagnosis and combined therapy are the keys to achieve good curative effects. This paper reviewed the recent progress in the main mechanisms,clinical manifestations,diagnosis and therapy of PE.

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 dynamic changes of TNF-?/IL-10in model rats with acute necrotizing pancreatitis (ANP)and to study the intervention outcome of salvia miltiorrhiza.METHODS:A total of96rats were randomly divided into three groups(each with32rats):pancreatitis(P)group,salvia miltiorrhiza treatment(T)group and control group(C),the blood samples of rats in each group were taken to determine levels of TNF-?and IL-10and the ratio changes of TNF-?/IL-10.Meanwhile,pancreas tissue sample was collected for pathological scoring.RESULTS:Group P had significantly higher serum levels of TNF-?and IL-10than did group C(P

Objective To investigate the macrophages (Mφ) phenotype mechanism in acute kidney injury caused by severe acute pancreatitis (SAP).Methods Sixty-four male Wistar rats were randomly divided into control group (SO) and SAP group (n =32 in each group).SAP rat model was made by retrograde cholangiopancreatic injection of 5％ sodium taurocholate.At 2,6,12 and 24 h after modeling,the samples of blood and kidney tissue were collected.The levels of blood urea nitrogen (BUN) and creatinine (Cr) were detected by using automatic biochemical analyzer.The expressions of IL-12,TNF-α,IL-10 and TGF-β mRNA of kidney tissue were detected by fluorescence quantitative polymerase chain reaction (QRT-PCR).The levels of CD68,iNOS and Arg-1 were measured by Western blot.Results In the SAP group at each interval,BUN and Cr concentrations were significantly higher than those of the control group (P ＜ 0.01,P ＜ 0.05) ; Compared with the control group,the expressions of IL-12,TNF-α,IL-10 and TGF-β mRNA in renal tissue of SAP group were significantly higher (P ＜ 0.01,P ＜ 0.05).In the SAP group,the levels of CD68,iNOS and Arg-1 were higher than those in the control group.Conclusions Inflammation and inflammatory imbalances may be pathological factors of acute kidney injury following SAP.

Objective To investigate the protective effect of lipsomal clodronate against hepatic injury in rats with acute necrotizing pancreatitis (ANP). Methods 48 SD rats were randomly divided into control group, ANP group and lipsomal clodronate group, respectively. The models of ANP were established by injection of sodium taurocholate into the pancreatic capsule. Lipsomal clodronate was prepared by means of thin film. Blank liposomes and clodronate-containing liposomes was injected via caudal vein in ANP group and lipsomal clodronate group, respectively. The rats were sacrificed at 2, 6 h after ANP induction, the serum levels of ALT, AST and AMS, IL-6,IL-12 were measured, and pathologic changes of liver and pancreas were observed. Results At 6 h, serum level of ALT was (73 ± 11) U/L, (257 ± 33) U/L and (184 ± 29) U/L in control group, ANP group and lipsomal clodronate group, respectively;serum levels of AST were (190 ± 32)U/L, (590 ± 70)U/L and (430±52)U/L, respectively;serum levels of AMS were (814±80)U/L, (5031 ± 471) U/L and (2843 ± 236) U/L, respectively, serum levels of IL-6 were (26.7 ± 5.7) pmol/L, (218.0 ±4.7)pmol/L and (112.3 ± 8. O) pmol/L, respectively;serum levels of IL-12 were (4. 2 ± 1.0) pmol/L,(309.5 ± 8.5) pmol/L and (153.7 ± 6.3) pmol/L. The values in ANP group and lipsomal clodronate group were significantly higher than those in control group, while the values in lipsomal clodronate group were significantly lower than those in ANP group (P < 0. 01). Pathologic changes of liver and pancreas were significantly attenuated in lipsomal clodronate group. Conclusions Intravenous liposomal clodronate could exert protective effects on the hepatic injury in rats with ANP.

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 protective effects of lipsomal clodronate on renal injury in rats with severe acute pancreatifis and the assessment of renal injury. Method Totally 48 rats were randomly divided into three group:normal control group (C);SAP group, in which rats were treated with pure liposomal (P);treatment group, in which SAP rats were treated with liposomal clodronate disodium(T). The SAP model of rat was induced by injection of 5 % sodium taurochohte beneath the pancreatic membrane. Rats of normal control group received isovolumetric injections of 0.9% physiological saline solution instead of sodium taurocholate. Blood samples were collected to measure AMS,BUN,Cr,IL-6 and IL-12 at 2 hors, 6 hours after SAP. At the same time, the samples of pancreatic and renal tissues were taken for observing the pathological changes. Results Compared with controlgroup, serious renal and pancreatic damages were found in group P, and the AMS, BUN, Cr levels elevated signifi-candy (P < 0.01). Compared with group P,the renal and pancreatic damages were attenuated in group T, and the levels of Cr and AMS decreased significantly (P < 0.01), and the IL-6, IL-12 were decreased at 2 hours and 6 hours (P < 0.01). The BUN decreased significantly at6 hours (P < 0.05). Conciusions Excessive release of inflammatory mediator play an important role in renal injury in SAP. Lipsomal clodronate disodium can alleviate the damage of pancreas and kidney.

Objective To investigate the protective effect of clodronate SPIO liposomes on liver injury of rats with severe acute pancreatitis(SAP)and the role of MRI in evaluating the extent of liver injury.Methods Superparamagnetic Fe3O4 nanoparticles were prepared by chemical coprecipitation.Clodronate-SPIO-containing liposomes was prepared by the thin-film method.SAP models were prepared by a uniform injection of sodium taurocholate(2 ml/kg body weight)into the subcapsular space of the pancreas.SD rats were randomly divided into control group,SAP plus SPIO group, and clodronate-SPIO-containing liposome group.Six hours after SAP models were available,T2-weighted MRI scanning(in the same plane)of the liver of rats in each group were performed.At the end of the scanning,blood samples were taken from the supcrior mesenteric vein to measure the contents of serum ALT and AST.Meanwhile, The pathological changes in the liver and pancreas were observed.Results Transmission electron microscopic examination showed that liposomes had a uniform size.No changes in the pancreas of rats in control group were noted.The pathological changes in the pancreas and liver of rats in SAP plus clodronate-SPIO-containing liposome group were significantly milder than those in SAP plus SPIO liposome group.The contents of serum ALT and AST in rats in SAP plus SPIO liposome group were significantly higher than those in control group(P＜0.01), while the contents of serum ALT and AST in rats in SAP plus clodronate-SPIO-containing group were significantly lower than those in SAP plus SPIO liposome group(P＜0.01).The MRI signal intensity of the liver in SAP plus SPIO liposome group and SAP plus clodronate-SPIO-containing liposome group was significantly lower than that in control group.The significant changes in the MRI signal intensity of the liver in SAP plus SPIO liposome group and SAP plus Clodronate-SPIO liposome group were noted(P＜0.01).Conclusion Clodronate-containing liposomes have protective effects against liver injury in SAP rats and SPIO can be used as a tracer for MRI examination.