Anti-inflammatory effect of interleukin-35 in mice with colitis and its mechanism / 浙江大学学报·医学版

OBJECTIVE@#To investigate the anti-inflammatory effect and mechanisms of interleukin-35 (IL-35) in inflammatory bowel disease.@*METHODS@#BALB/c mice were divided into three groups with 10 mice in each group:control group, model group (oral administration of 4% glucan sodium sulfate for 7 d) and IL-35-treated group (oral administration of 4% glucan sodium sulfate for 7 d, intraperitoneal injection of 2 μg IL-35 at d2-5). Disease activity index (DAI) was scored every day. After 7 d, the mice were sacrificed, and the serum and intestinal tissue samples were collected. The gross morphology of the colon was observed; HE staining was used to observe the pathological changes of colon tissue; flow cytometry was employed to detect the change of macrophage polarization ratio in colon tissue; the mRNA expression levels of cytokines IL-6, TNF-α, IFN-γ, IL-10 and SHIP1 in colon tissue were determined by real-time quantitative RT-PCR; the expression and distribution of SHIP1 in colon tissue was measured by immunohistochemistry; Western blotting was adopted to detect the expression level of SHIP1 protein in colonic intestinal tissues of each group.@*RESULTS@#The DAI scores of the mice in the model group were higher than those in the control group, while the DAI scores in the IL-35-treated group were lower than those in the model group (all 0.05). Compared with the model group, microscopic inflammatory infiltration score was decreased and microscopic crypt destruction and score was significantly lower in IL-35-treated group (all <0.05). The relative expression of proinflammatory cytokines IL-6, TNF-α and IFN-γ in the colon tissue of IL-35-treated group was decreased compared with the model group, while the relative expression of IL-10 mRNA was higher than that of the model group (all <0.05). Compared with the control group, the proportion of M1 macrophages in the model group increased (<0.05), while the proportion of M1 macrophages in the IL-35-treated group was lower than that in the model group (<0.05). The relative expression of SHIP1 mRNA and protein in the colon tissue of IL-35-treated group was higher than that in the model group (all <0.05).@*CONCLUSIONS@#IL-35 can inhibit the polarization of M1 macrophages and regulate inflammatory cytokines to promote anti-inflammatory effect on mice with colitis.

Alteration of C/EBPs and endoplasmic reticulum stress related molecules expression in pancreatic tissues in rats with hypertriglyceridemia related acute pancreatitis / 中华胰腺病杂志

Objective To investigate the alteration of C/EBP α,C/EBP β and endoplasmic reticulum stress (ERS) related molecules (IRE1α and sXBP1) expression in pancreatic tissues in rats with hypertriglyceridemia related acute pancreatitis.Methods Ninety six Sprague Dawley (SD) rats were divided into 4 groups:control group,hypertriglyceridemia (HTG) group (n =24,fed with high fat diet for 2 weeks),AP group (n =24),HTG + AP group (n =24),and AP was induced by peritoneal injection of cerulein.The rats were sacrificed at 3,6,9,24 h after AP induction,respectively.The pathological changes of the pancreatic tissues were observed and scored by HE staining.Plasma levels of IL-1β,IL-6 and TNF-α were measured by ELISA.The expression of IRE1α,sXBP1,C/EBPoα,C/EBPβ mRNA were analyzed by real time PCR.The expressions of IRE1α,sXBP1,NF-kB,C/EBPα and C/EBPβ protein were determined by Western Blot.The expressions of C/EBPα and C/EBPβ proteins were also determined by immunohitochemistry.Results After two weeks of high fat diet,serum levels of triglyceride(TG) and total cholesterol (TC) in HTG group,HTG + AP group were much higher than those of the control group,and the difference was statistically significant (P ＜ 0.05).The pancreatic tissue injury was more severe in HTG + AP group,particularly at 9 h (P ＜ 0.05).And plasma IL-1β,IL-6 and TNF-α levels were also much higher in HTG + AP group when compared with that of AP group,the differences were all significant at 9 h (P=0.011;P=0.034;P =0.027).After AP induction,IRE1,sXBP1,C/EBP and C/EBPβ mRNA began to be up-regulated at 3 h,and IRE1 mRNA reached the highest level at 24 h,sXBP1 mRNA at 9 h,while C/EBP and C/EBPβ mRNA reached the highest level at 6 h.Compared with AP group,IRE1,sXBP1,C/EBP and C/EBPβ mRNA levels were much higher in HTG + AP group.In addition,as to IRE1 and sXBP1 mRNA,the difference was significant at 3,6,9,24 h,and C/EBP mRNA at 6,9,24 h,C/EBPβ mRNA at 6 and 9 h (P ＜ 0.05).After AP induction,IRE1α,sXBP1 and NF-kB proteins in the pancreatic tissue began to be up-regulated at 3 h,and all reached the highest level at 9 h.IRE1α,sXBP1 and NF-kB proteins were up-regulated more obviously in HTG + AP group,and the up-regulation in HTG + AP group was higher than that in AP group,and the high expressions of C/EBPα and C/EBPβ proteins could only be detected at 6 and 9 h in the HTG + AP group,while there was no expression detected in AP group.Conclusions C/EBPα,C/EBPβ,IRE1α and sXBP1 may be involved in the pathogenesis of HTG related AP,and IRE1α/sXBP1 pathway and C/EBPoα,C/EBPβ may mediate the pathologic injury and inflammation process of HTG related AP.

Role and Therapeutic Prospect of Intestinal Microecology in Crohn’s Disease / 胃肠病学

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.

Research on the hospital management system in view of total quality management / 中华医院管理杂志

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.

Expression and its clinical significance of CD9 protein in pancreatic cancer / 中华消化杂志

Objective To examine the expression of CD9 protein in pancreatic cancer tissues and adjacent tissues,and to analyze its relation with the progress and prognosis of pancreatic cancer.Methods From September 2005 to December 2009,surgical resected cancer tissues and adjacent tissues of 90 patients with pancreatic cancer and their clinical data were collected.The expression of CD9 protein in pancreatic cancer tissues and adjacent tissues was detected by immunohistochemistry,and its relationship with clinicopathological features was analyzed.Chi-square test was performed for comparison of ratios between groups.Overall survival (OS) analysis of 90 patients after surgery was performed.Results The high expression rate of CD9 protein (64.4％,58/90) in cancer tissue was significantly higher than that in cancer adjacent tissue (45.6％,41/90),the difference has statistically significant (χ2 =6.847,P＜0.05).CD9 protein was highly expressed in most of pancreatic cancer tissue which was well differentiated or without lymph node metastasis (74.6％ (50/67) vs 39.1％ (9/23),χz =9.554,P＜0.01; 50.0％(17/34) vs 73.2％(41/56),χ2 =5.856,P＜0.05 respectively).However,the expression of CD9 was not correlated with gender and age (both P＞0.05).OS and progression-free survival (PFS) of the patients with CD9 highly expressed were significantly longer than those with low expression of CD9 (median OS:33.0 months vs 7.0 months,χ2 =15.400 P＜0.01.Median PFS:30.5 months vs 5.0 months,χ2 =13.750,P＜0.01).Conclusion CD9 protein is a kind of protein related with the invasive ability of pancreatic cancer,which may play a role in progression and metastasis of pancreatic cancer and can help to determine the prognosis to a certain extent.

Effect of pancreatic stellate cells on invasion and metastasis of human pancreatic cancer cell line AsPC-1 / 中华胰腺病杂志

Objective To investigate the effect of pancreatic stellate cells on invasion and metastasis of human pancreatic cancer cell AsPC-1,and to determine the role of SDF-1 in this process.Methods PSCs were routinely isolated and cultured,and PSCs conditioned media(PSC-CM) was collected and concentrated.Different concentrations of PSC-CM,anti SDF-1 and their combination were used to treat AsPC-1 cells,and MTT assay was applied to detect the proliferation of pancreatic cancer cells.Transwell chamber migration assay was employed to detect the migration of AsPC-1 cells.In vitro invasion assay was used to determine the invasion of AsPC-1 cells.Results A490 values of AsPC-1 cell in control group and 0.25,0.5,1 μg/lμl PSCCM group were 0.437 ±0.041,0.472 ±0.048,0.553 ±0.057,0.690 ±0.051,and PSC-CM promoted cell proliferation in a dose dependant manner.The difference between 0.5,1 μg/μl PSC-CM group and control group,and between 1 μg/l and 0.5 μg/μl PSC-CM group was statistically significant (P＜0.05).A490 values of control group,anti SDF-1 group,PSC-CM group and PSC-CM ± anti SDF-1 group were 0.407 ±0.028,0.416 ±0.030,0.629 ±0.048,0.481 ±0.049.The numbers of penetrating cells were 35.3 ±7.1,34.8±5.6,140.9 ± 12.7,56.5±5.9,and the numbers of invasive cells were 27.1 ±2.9,29.1 ±4.2,81.5 ±8.2,46.4 ± 4.4.The difference between anti SDF-1 group and control group was not statistically significant.The proliferation,migration and invasion of pancreatic cancer cells in PSC-CM group was significantly higher than those of control group (P ＜0.05 or P ＜0.01).The proliferation,migration and invasion of pancreatic cancer cells in PSC-CM ± anti SDF-1 group was significantly lower than those of PSC-CM group,but they were significantly higher than those of control group (P ＜ 0.01).Conclusions PSCs can promote proliferation,migration and invasion of pancreatic cancer cells AsPC-1,and the mechanism may be partly due to SDF-1/CXCR4 receptor ligand axis.

Effect of intraductal ultrasonography in the diagnosis of non-opaque choledocholith / 中华消化杂志

Objective To evaluate the diagnostic value of intraductal ulstrasonography(IDUS)in non-opaque bile duct stones.Methods Between January 2009 and August 2010 in the Department of Gastroenterology at Shanghai 10th People's Hospital,a total of 183 patients(male:102 cases,mean age 69 years； female:81 cases,mean age 71 years)were enrolled,who were suspected of bile duct stones or stenosis which could not be diagnosed by abdominal CT,MRI,and abdominal B-mode ultrasound.All the patients underwent endoscopic retrograde cholangiopancreatography(ERCP)first,and then patients with non-opaque bile duct stones followed by IDUS.Results A total of 134 cases (73.2％)of bile duct stones were diagnosed by ERCP,49 cases(26.8％)were negative.And then the 49 patients underwent IDUS,of whom 24 patients with sand-like stones,11 patients with lowdensity stones,6 patients with ampullary cancer,2 patients with pancreatic cancer,6 patients with sclerosing cholangitis.The diagnostic accuracy of IDUS in the position and quality of bile duct stones was 100％,higher than that of ERCP,which was 80％.After ERCP,pancreatitis occurred in 3 patients and improved after conservative treatment,there was no complications like perforation and bleeding.Conclusion The diagnostic accuracy of IDUS in the position and quality of bile duct stones is high,which can make up for the misdiagnosis by ERCP without increasing the complications.IDUS can provide reliable basis for the diagnosis of clinical bile duct stones.

Inhibitory effect of vasostatin on migration of pancreatic cancer endothelial cells / 中华胰腺病杂志

ObjectiveTo investigate the effect of vasostatin on the migration of pancreatic cancer endothelial cells.Methods Ad-vasostatin with different concentrations of vasostatin was used to transfect pancreatic cancer endothelial cells.Ad-LacZ transfection and PBS was used as control.The effect of vasostatin gene mediated by adenovirus on the migration of pancreatic cancer endothelial cells was measured by woundhealing assay,transwell migration assay,and tube formation assay.ResultsThe scratched lines in PBS group and Ad-LacZ group were almost healed 48 hours later,while the lines in Ad-vasostatin group were rarely healed.At the MOI of 1,2,5,the migration rate of Ad-Laz group was ( 84.7 ± 2.6) ％,(80.7 ± 1.7 ) ％ and (81.3±4.0)％,while the corresponding values were (77.7 ±2.1)％,(67.3 ±2.1)％ and (38.8 ±2.1 ) ％ in Ad-vasostatin group.Transwell migration assay indicated that the number of migrated cells in Advasostatin group was inhibited in a dose-dependant manner,at the MOI of 5,the migration became significantly decreased (F=180.88,P ＜0.05).At the MOI of 1,5,10,the number of tubes in Ad-LacZ group was 118±6,120±6 and 82±5,while the corresponding values were 65±4,21±4 and 4 ±1 in Ad-vasostatin group.The number of tubes of pancreatic cancer endothelial cells was inhibited by Ad-vasostatin in a dose-dependant manner,at the MOI of 10, it was difficult to form the tubes (F-300.85,P＜0.05). Conclusions The vasostatin gene mediated by adenovirus has a significant inhibitory effect on the migation of pancreatic cancer endothelial cells in vitro in a dose-dependent manner.

A multi-center retrospective study on the judgment value of bedside index for severity in acute pancreatitis / 中华消化杂志

Objective To compare the value of bedside index for severity in acute pancreatitis (BISAP),Ranson score and Balthazar computed tomography severity index (CTSI) in predicting the severity and prognosis of acute pancreatitis (AP).Methods From 2005 to 2011 in Shanghai,the clinical data of 1004 AP cases from seven hospitals was collected and retrospectively analyzed.The value of BISAP score,Ranson score and Balthazar CTSI in predicting the severity and prognosis of AP were assessed with receiver operator characteristic (ROC) curve.Results Among 1004 patients,the main cause of AP was biliary disease (580 cases),about 57.77％.The incidence of pancreatic necrosis,mortality and SAP increased along with BISAP score.The risk of pancreatic necrosis in patients with CTSI ≥ three was significantly higher than that of ＜ three.The risk of pancreatic necrosis and SAP in patients with BISAP score ≥ two was significantly higher than that of ＜ two (OR:4.93,95％CI 3.62-6.70; OR 2.62,95％CI 1.59-4.31,respectively).There was no significant difference in the accuracy of predicting the progression and mortality of AP among these three score systems.However the sensitivity of BISAP score (OR:61.54,95％CI 35.09-87.99) in predicting the progression and mortality of AP was better than that of Ranson (OR:46.15,95 ％ CI 19.05-73.25) and CTSI (OR:46.15,95％CI 19.05-73.25).Conclusions BISAP score is easy to perform and when combined with CTSI,it helps to make the diagnosis and classification of AP in time,predict the prognosis accurately.Compared with Ranson score,BISAP score has higher clinical value.

Study on the biological characteristics of pancreatic cancer vascular endothelial cells / 中华消化杂志

Objective To explore the biological characteristics of pancreatic cancer vascular endothelial cells,including the aspects of morphology,species,genetics,vascular formation ability,and proliferation ability in vitro.Methods The human pancreatic cancer cells were inoculated in nude mice pancreas to get pancreatic cancer vascular endothlial cell.The pancreatic cancer vascular endothelial cells were cultured in vitro.The passage number and passage time were recorded.The morphological features under common microscopy of each passage were observed.The species origin and genetic characteristics of the pancreatic cancer vascular endothelial cells were detected by karyotype assay.The ability of angiogenesis of the pancreatic cancer vascular endothelial cells in vitro was determined by tube formation assay.The proliferation of the pancreatic cancer vascular endothelial cells in vitro was measured by MTT method.The data were analyzed by one way analysis of variance and paired difference test.Results Under appropriate culture condition, the pancreatic cancer endothelial cells were passaged every two to three days.Once confluence was attained,the cells were in monolayer growth and with cobblestone feature.The species type of the pancreatic cancer vascular endothelial cells was human type.A large number of polyploid cells, non-integer multiple chromosomes cells, nuclear chromosome loss, nuclear chromosome dislocation, and unanalyzable fragments were observed.The pancreatic cancer vascular endothelial cells could form a hollow tubular structure in vitro.After cultured for 48 and 72 hours,the absorbance of the pancreatic cancer vascular endothelial cells was 0.581 ± 0.014 and 1.082 ± 0.033 respectively,both were significantly higher than those of human umbilical vein vascular endothelial cells (0.379± 0.038,t=8.720,P=0.001;0.604±0.026,t=19.883,P＜0.01).Conclusions The species origin of pancreatic cancer vascular endothelial cells is same as human pancreatic cancer cells.The cells have typical morphological features and in vitro angiogenesis formation ability of vascular endothelial cells,whose genetic feature is instable and proliferation is active.

Expression of aquaporins-4 in brain tissue of rats with pancreatic encephalopathy / 中华胰腺病杂志

Objective To study the expression of aquaporins-4 (AQP4) in the brain tissue of rats with pancreatic encephalopathy (PE) induced by phospholipase A2 and to explore the role of aquaporins-4 in PE.Methods Twenty five healthy Wistar rats were randomized into 3 groups:blank group ( n =5),PE group (n =10 ) and control group (n =10 ).The experimental model was established in rats by injecting phospholipase A2 into carotid artery (0.1 ml/100 g body weight).Same amount of normal saline was used in the control group and no treatment was used in the blank group.One day later,the rats were sacrificed,then the measurement of brain tissue wet/dry (W/D) weight ratio was performed,and brain tissue was routinely pathologically examined,immunohistochemistry and Western blotting were performed in each group to detect the expression of aquaporins-4.Results There was no obvious brain tissue pathological change in the control group and blank group.Neurons in the brain tissue of PE rats presented with significant edema and ballooning degeneration,infiltration of inflammatory cells,leukocyte aggregation around the microvessels.The water contents in the brain tissue in the blank group and control group,PE group were (61.44 ±0.36)％,(63.20±0.32)％ and (78.33 ±0.24)％,and it was significantly higher in PE group than that in the control and blank group (P＜0.05).The expressions of aquaporins-4 in the brain tissue were 0.41 ±0.27,0.49 ±0.13,0.98 ±0.21,respectively,and it was significantly increased in PE group than that in the control and blank group (P ＜ 0.05 ).Conclusions Aquaporins-4 may play important roles in the pathogenesis of pancreatic encephalopathy.

Expression and methylation of Iroquois homeobox protein 1 in pancreatic cancer / 中华胰腺病杂志

Objective To investigate the expression of Iroquois homeobox protein 1 ( IRX1 ) gene and the hypermethylation status of its promoter in pancreatic cancer,and their relationship.Methods Real-time PCR was used to quantitatively detect IRX1 gene expression level of 12 sets of resected pancreatic cancer tissue and 6 sets of pancreatic cancer cell lines; gene sequences analysis was used to detect the structure of IRX1 promoter; DNA methylation inhibitor 5-Aza-2′-deoxycytidine (5-Aza-dC) was used in pancreatic cancer cell lines,and then the methylation of IRX 1 was measured by methylation-specific PCR (MSP) and unmethylation sequence-PCR (USP) methods.Results Expression of IRX1 mRNA in pancreatic cancer tissue was 0.31 ± 0.11,which were significantly lower than that in normal pancreatic tissue ( 1.05 ±0.32,P ＜0.01 ).IRX1 mRNA expression of AsPCl,BxPC3,Capan 2,PANCl,PaTu8988 and SW1990 were 0.36 ± 0.08,0.34 ±0.16,0.37 ±0.11,0.25 ±0.06,0.31 ±0.04,0.36 ±0.02,which were significantly lower than that in human kidney epithelial 293 cells ( 1.03 ± 0.28,P ＜ 0.05 or ＜ 0.01 ).Analysis of IRX1 gene sequence showed abundant CpG islands in promoter.Hypermethylation of IRX1 promoter site was found in all pancreatic cancer cell lines.However,its methylation status could be reversed by 5-Aza-dC,and the IRX1 expression was also restored.Conclusions IRX1 mRNA expression is down-regulated in pancreatic cancer,and it is related with promoter CpG islands hypermethylation.

Involvement of pancreatic beta cell in pancreatic regeneration following experimental acute pancreatitis / 中华胰腺病杂志

Objective To investigate the role of pancreatic β cell on pancreatic regeneration following experimental acute pancreatitis.Methods Eighty-seven SD male rats were randomly divided into four groups:control group ( n =15 ),STZ group ( n =24),L-Arg group ( n =24 ),STZ + Arg group ( n =24).60 mg/kg of STZ was administrated by intraperitoneal injection to induce the diabetes model.2.5 g/kg body weight of LArg was administrated by intraperitoneal injection to induce the acute pancreatitis model.The rats were sacrificed 1,3,5,7 d later and the serum levels of amylase and glucose were measured.Relative pancreatic weight (pancreatic weight/body weight) were measured.Pancreatic tissue underwent routine pathologic examination,and the percentage of area of necrosis and tissue transformation was calculated.The expression of Reg4 and insulin was performed by immunofluorescence.Results Serum level of glucose significantly increased after STZ injection.After L-Arg injection,serum level of amylase significantly increased,and there was pancreatic tissue edema,necrosis,infiltration of inflammatory cells,which suggested the successful model induction.The percentage of area of necrosis in STZ + L-Arg group was (71.6 ± 6.0) ％ at the 3rd day,which were significantly higher than (42.3 ± 4.0 ) ％ in L-Arg group; the percentage of area of transformation was (45.6 ± 5.4) ％,which were significantly lower than (78.5 ± 6.4) ％ in L-Arg group.Expression of Reg4 in pancreatic islets of STZ + L-Arg group was significantly lower than those in L-Arg group.Conclusions STZ impairs pancreatic β cells,aggravates pancreatic damage following L-arginine induced pancreatitis and inhibits pancreatic regeneration.

The effects of enteral nutrition versus total parenteral nutrition on gut barrier function in severe acute pancreatitis / 中华内科杂志

Objective To evaluate the effect of enteral nutrition (EN) versus total parenteral nutrition(TPN) on gut barrier function in patients with severe acute pancreatitis (SAP). Methods Sixtythree patients with SAP enrolled from 4 hospitals were randomly assigned into EN group(29 cases) and TPN group(34 cases). EN group patients were fed via a spiral nasojejunal feeding tube placed routinely by endoscopy or fluoroscopy, and TPN group patients were nourished intravenously with TPN during the same period. The changes of serum endotoxin, diamine oxidase, and urinary excretion of lactulose and mannitol ratio (L/M) were observed. Results Plasma concentration of endotoxin were markedly decreased in EN group as compared with that in TPN group at the 7th,14th ,21th day of entry trial [(39. 30 ± 15. 82) EU/L vs (73.05 ±21.16) EU/L,(22.64 ±14.31) EU/L vs (49.34 ±24.54) EU/L,(14.81 ± 10.93)EU/L vs ( 30. 08 ± 14. 10 ) EU/L, P ＜ 0. 05]. Plasma concentration of diamine oxidase were markedly decreased in EN group as compared with that in TPN group at the 7th, 14th day of entry trial [(9. 97 ± 3. 84)U/Lvs (19.89±9.89)U/L,(5.42±1. 84) U/Lvs (8.79 ±4.08) U/L, both P ＜ 0. 05]. The urinary L/M decreased significantly in EN group than those in TPN group at the 7th, 14th,21th day of entry trial (0.28 ±0.25 vs 0. 65 ±0.45,0.21 ±0. 18 vs 0.54 ±0.41,0.08 ±0.04 vs 0.29 ±0.06, all P＜0.05).Conclusion EN has better effect on improving intestinal barrier function than TPN in treatment of patients with SAP.

Association of serum cytokines with pancreatic function in patients after acute pancreatitis / 中华消化杂志

Objective To investigate the effect of cytokines on pancreatic function in patients after acute pancreatitis(AP) and its mechanisms. Methods Fifty-nine patients (mild in 25 and severe in 34) after AP and 20 healthy controls were enrolled in the study. Serum levels of cytokines including hepatocyte growth factor (HGF), epidermal growth factor(EGF), basic fibroblast growth factor (bFGF), regeneration protein(Reg)-1 and Reg-4 were determined using enzyme-linked immunosorbent assay (ELISA). Fasting blood-glucose, insulin, C-peptide and fecal elastase 1 (FE1) were detected for evluation of endocrine and exocrine pancreatic function. The association of pancreatic function with clinical parameters and serum cytokines was analyzed. Results The expression of FE1 was lower in patients [(205.9±18.3) μg/g] after AP in comparison with the controls [(333.9±19.7) μg/g, P<0. 01], but levels of fasting blood-glucose, C-peptide and insulin were higher in patients group (P<0.01). Serum level of HGF was higher in patients with insufficient pancreatic exoerine [(983.76±372.65) pg/ml] than those with normal exocrine function [(263.44±110. 35) pg/ml]. Meanwhile,EGF level was higher in patients with DM after AP [(704.41±190. 37) pg/ml] than those without DM [(360. 03±48.39) pg/mh P<0.05]. There was a negatively correlation between FE1 and HGF (P ＜0. 01). The abnormal fasting blood glucose was correlated with CT grading (P<0. 05).Conclusions The patients after AP develope insufficient exocrine and endocrine function. Serum EGF and HGF may be associated with restoration of pancreatic endocrine and exocrine function.

Effect of AMD3100 on the proliferation and angiogenesis of AsPC-1 cells / 中华胰腺病杂志

Objective To investigate the effects of blockade on non-peptide specific SDF-1/CXCR4 receptor ligand system with AMD3100 on the proliferation and angiogenesis of human pancreatic cancer cells AsPC-1. Methods AsPC-1 was divided into control group, SDF-1α group, group, SDF-1α + AMD3100 group. MTT test was performed to determine the proliferative level of AsPC-1 cells. Vascular endothelial growth factor (VEGF) was detected with Western blotting assay. Immunohistochemistry was used to detect the microvessel density (MVD) in subcutaneous xenografts of AsPC 1 of nude mice model, which was intratumorally and peritumorally injected with AMD3100. Results SDF-1α could induce the proliferation of AsPC-1(1.430 ±0. 122 vs 1. 002 ± 0. 001, P <0. 05). While the proliferative effect induced by SDF-1α could be inhibited by AMD3100 (0.983 ±0. 068vs 1.430 ± 0. 122, P <0.05). SDF-1α could induce the expression of VEGF (0. 565 ± 0. 047 vs 0. 439 ± 0.034, P < 0.05). While the protein expression of VEGF induced by SDF-1α on AsPC-1 cells was inhibited by AMD3100 (0. 450 ± 0. 071 vs 0. 565 ± 0. 04, P <0. 05). The growth and angiogenesis of subcutaneous xenografts of nude mice model were inhibited by AMD3100; the tumor inhibitory rate was 59. 5% at 24th day. The MVD of xenografts was significantly decreased (28.56 ± 6.94 vs 98.75 ± 20. 60, P < 0. 01 ). Conclusions AMD3100 could inhibit the proliferation and angiogenesis of AsPC-1 cells both in vitro and in vivo.

Effect on blockade of MCP 1 in early course of experimental acute necrotizing pancreatitis / 中华胰腺病杂志

Objective To investigate the potential role of MCP-1/CCL2 in experimental acute necrotizing pancreatitis (ANP) and complications. Methods 60 SD male rats were randomly divided into 3 groups: sham operation group ( n = 20 ), ANP group ( n = 20 ) and MCP-1 group ( n = 20 ). ANP model was induced by retrograde infusion of 3.5% sodium taurocholate, MCP-1 group received subcutaneous injection of MCP-1 antibody 0 h and 6 h after ANP induction. The serum levels of amylase, MCP-1, D-lactic acid,histological changes and the expression of MCP-1 mRNA of lung, small intestine and pancreas, the expression of MCP-1 protein in pancreas, MPO levels of small intestine MPO were determined. Results The serum levels of amylase, MCP-1, D-lactic acid in MCP-1 group at 12 h were (4666 ±412)U/L, (39.53 ±8.25)pg/ml and (6.3 ±2.2)mg/L, which were significantly lower than those in ANP group [ (9611 ±363)U/L, (63.42 ±9.32) pg/ml, (9.3 ± 2. 1 ) mg/L, P< 0.05 ) ]; the expression of MCP-1 mRNA in pancreas, small intestine and lung were 0.431 ± 0.009, 0. 211 ± 0.018 and 0.442 ± 0.017, which were significantly lower than those in ANP group [ (0.624 ±0. 010, 0. 523 ±0. 019 and 0. 569 ±0. 024, P <0.05) ]; the expression of MCP-1 protein in pancreas was 2.0 ± 0. 1, which was significantly lower than that in ANP group (4. 0 ± 0. 2, P <0.05). Lung and small intestine MPO were (11.1 ±3.0)U/g and ( 19.2 ±2.0)U/g, which were significantly lower than those in ANP group[(39.2±3.1)U/g and(13.1±2.1)U/g, P<0.05]. Conclusions Early blockade of MCP-1 not only attenuates the severity of ANP, but also decreases the degree of acute lung injury and intestine barrier dysfunction.

Effect of lipases on hyperlipidemic acute necrotizing pancreatitis / 中华胰腺病杂志

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.

Metabonomics study of urine samples of patients with hyperlipidemic pancreatitis / 中华胰腺病杂志

Objective Metabonomics method based gas chromatography-mass spectrometry (GC/MS)were used to analyze the urine samples of patients with hyperlipidemic pancreatitis (HLP) to describe the characteristics of metabolism changes of HLP,identify potential biomarkers,and investigate the role of metabonomics study in the management of AP.Methods 24 patients of HLP and 40 age,sex matched volunteers were enrolled and their urine samples were collected.The urine samples underwent preparation,derivation and GC/MS analysis,Orthogonal-Projection to Latent Structures-Discriminant Analysis (OPLS-DA)were performed to detect the metabolic profile difference between the HLP and control group.Results HLP patients can be precisely distinguished from healthy controls.21 metabolites (credibility ＞ 700 ) were identified using the reference compounds available in the libraries of NIST and Wiley.It was identified that levels of nicotinic acid,aconitie acid,citric acid,hippurie acid,hydroxyphenylacetic acid,hydroxyphenylpropionicacid were decreased,while the levels of tryptophan,tyrosine,tyramine,16-hexadecanoic acid,18octadecanoie acid were increased.It was also suggested that there was change in tricarboxylic acid cycle and gut bacterial flora,as well as fat metabolism and metabolism of amino acid.Conclusions There are differences between healthy controls and HLP patients in the term of GC/MS metabolic profiling,and the biomarkers in the metabolites could be found through metabonomics analysis,and the mechanisms of the metabolic changes could be explored.It was noted that the research of metabolites in the urine samples may be a useful tool to help diagnose and understand the pathogenesis of HLP.Metabonomics analysis is a promising research method.

The role of Hedgehog pathway in hepatic fibrosis and hepatic stellate cell activation / 中华消化杂志

Objective To investigate the role of Hedgehog pathway in hepatic fibrosis and its association with activation of hepatic stellate cells. Methods Twenty male Spragur-Dawley rats were divided into control and model groups with 10 each. The animal models were induced by injection with CCl4 and fed with fat-rich diet. The rats in both groups were sacrified at the 8 week with 5 each and the liver tissues were removed for HSC-T6 culture. The deposition of collagen fiber in liver was detected with HE and Masson staining. RT-PCR was used to detect the expressions of Sonic hedgehog (Shh), smoothened (Smo), patched (Ptc), Gli-1 and α-smooth muscle action (α-SMA) mRNA in HSC-T6 and liver tissues. The influence of cyclopamine (Cyc) and lipopolysaccharide (LPS) on HSC-T6 proliferation were assayed by MTT. The expressions of Shh, Smo, Ptc, Gli-1 and α-SMA mRNA after intervention with Cyc (100μmol/L) and LPS were measured by real-time PCR. Results A lot of lipo and collagen deposited in liver of model rats. The Shh,Smo,Gli-1 and α-SMA mRNA were highly expressed in model rats than those in control group (2-△△Ct were 20.45±3.31 vs. 1, 12.78 ± 0. 53 vs. 1, 10.88 ± 2.41 vs. 1, 4.91 ± 2. 59 vs. 1, respectively, all P value <0. 05). In vitro Cyc inhibited HSC-T6 proliferation in dose dependant manner (F=636.81, P<0.01). Compared to the control group, the mRNA expressions of Smo, Ptc, Gli-1,α-SMA in HSC-T6 were significantly reduced after Cyc intervention (2△△Ct, were 0. 20±0. 11, 0. 21 ± 0. 08, 0. 28 ± 0. 05,0. 27±0.10,respectively, all P values<0.01). Conclusion The expression of members of Hedgehog pathway are increased in the progress of hepatic fibrosis, which may accelerate the hepatiee fibrosis by activating HSC.