Objective To study the influence on heat shock protein 27 (Hsp27) and tumor necrosis factor ? (TNF-?) by inhibiting ubiquitin proteasome system in cardiac ischemia-reperfusion (I/R) rats. Methods Left anterior descending (LAD) coronary artery was ligated for 30 min and then released for 6 h in adult Sprague-Dawley rats. Five minutes before reperfusion, MG-132 (N-benzoyloxycarbonyl (Z)-Leu-Leuleucinal) at dose of 0.75 mg/kg was given by vein injection in I/R+T group, but I/R group, I group and sham group were given the same volume of sterile saline. The changes of myocardial infarct size, cardiac protein and mRNA levels of Hsp27 and TNF-? were measured. Results Compared with I/R group, the myocardial infarct size was significantly reduced (P

Objective To explore the effects of rosiglitazone (ROSI),a peroxisome proliferator-activated receptors-gamma (PPAR-γ) ligand,on hyperlipidemia in rats with severe acute pancreatitis (SAP) associated with lung injury.Methods A total of 120 male SD rats received intragastric administration of high fat diet for two weeks to induce experimental hyperlipemia.The hyperlipidemic rats were randomly (random number) divided into six groups:hyperlipidemia (HL) group (n =20),hyperlipidemia with SAP (HP) group (n =20),hyperlipidemia with rosiglitazone intervention (HRP) group (n =20),hyperlipidemia with rosiglitazone and antagonist to rosiglitazone (HRGP) group (n =20),rosiglitazone control (HR) group (n =20) and antagonist control (HG) group (n =20).The SAP was induced by a retrograde infusion of 5％ sodium tauroholate into bile-pancreatic duct,and the SAP was established in HP group,HRP group and HRGP group.In HL group,HR group and HG group,equivalent volume of normal saline was used instead of sodium taurocholate.In HRP group and HR group,ROSI (6 mg/kg) was administered via the femoral vein 1 hour prior to the administration of sodium taurocholate.In HRGP group,GW9662 (0.3 mg/kg),an antagonist to PPRA-gamma,was given via the femoral vein 30 min prior to the administration of ROSI.In HG group,only GW9662 (0.3 mg/kg) was given via the left femoral vein 30 min prior to pretend SAP modeling.Rats from each group were sacrificed by exsanguination 12 h after SAP modeling.Blood samples were taken from all subjects to measure serum amylase (AMY),total cholesterol (TC),triglycerides (TG),Successive sections of the paraffin embedded tissue from pancreas and lung were taken for pathological examination with hematoxylin-eosin (HE) staining.Histopathological changes of pancreatic and pulmonary tissues observed under light microscope were evaluated.In pulmonary tissue,nuclear factor-kappa B (NF-κB) p65 expression was assayed by immunohistochemistry.Intercellular adhesion molecule (ICAM-1) protein and tumor necrosis factor-α (TNF-γ) protein levels were studied using Western blot analysis.Results The serum levels of TC and TG in HL group and HP group were significantly higher than those in HR group and HRP group (1.24 ± 0.28,1.14 0.08 vs.0.41 ±0.17,0.58±0.12;14.86±1.47,12.42±0.96 vs.6.52±2.04,7.36±0.95,allP＜ 0.05);The levels of serum AMY,W/D ratio,pancreas pathologic score,lung pathologic score,expression of NF-κB p65,ICAM-1 and TNF-α in pancreas in the HP group and HRGP group were significantly higher than those in HL group,HR group,and HG group (6 501.9 ±3 770.0,5 922.2 ±925.9 vs.1 139.3 ± 35.6,1 070.8 ±67.0,1 012.4 ±94.7;3.14±0.16,3.06±0.12vs.1.81 ±0.13,1.76±0.23,1.83 ± 0.18;all P ＜0.05);Compared with the HP group and HRGP group,the levels of serum AMY,TC and TG were significantly decreased in HR group and HRP group,ameliorating pancreas and lung pathological damage,and down-regulating the expression of NF-κB p65,ICAM-1 and TNF-α in pulmonary tissue (all P ＜ 0.05).While there were no statistically significant differences in above biomarkers between HP group and HRGP group (all P ＞ 0.05).Conclusions Our study demonstrates that ROSI exerts anti-hyperlipidemic effect and anti-inflammatory effect on hyperlipidemia in rats with sodium taurocholate-induced severe acute pancreatitis associated with lung injury by inhibiting NF-κB and down-regulating the expression of TNF-α and ICAM-1.

BACKGROUND: Vessel extracellular matrix (VECM) is a natural scaffold material obtained from vascular tissues, which can stimulate angiogenesis and accelerate vascularization of tissue-engineered graft, however, the mechanism is poorly understood.OBJECTIVE: To explore the vascularization effects of release of vascular endothelial growth factor (VEGF) from VECM in ureteral reconstitution.DESIGN, TIME AND SETTING: An in vitro cytology observation. The experiment was performed at the Biomedical Engineering Laboratory of First Clinical Medical Science College, Wuhan University, between April and August in 2009.MATERIALS: Abdominal aorta was obtained from 5 rabbits to prepare VECM.METHODS: The VEGF released from VECM in vitro was evaluated by ELISA, the effects of cell proliferation by the released VEGF was detected by MTT colorimetric assay. The defected ureters of rabbits were repaired by homologous VECM in vivo.Then the recovery of the defected ureters and the situation of vasculogenesis were detected at different time point.MAIN OUTCOME MEASURES: The detection of VEGF contents in VECM; and the effects of VECM on vascular endothelial cell proliferation and ureteral reconstitution.RESULTS: In vitro experiment presented that the peak amplitude concentration of VEGF released from VECM in PBS solution was (124.10±1.42) ng/L, which showed proliferative effect on vascular endothelial cells. In vivo, there were some blood vessels on the VECM at 2 weeks after implantation. Epithelial coverage was evident in the lumen of the marginal part of the VECM grafts and the smooth muscle extended from the transition zone. After 8 weeks, the quantity of the blood vessel was increased and the caliber of the blood vessels became wide. There was thickness epithelial lamina in the graft, and the muscle fibers had an organized spatial alignment, forming variably sized bundles. After 16 weeks, there were no significant differences between the regenerative tissue and the normal tissue in morphology.CONCLUSION: The homologous VECM can release VEGF when implanted as tissue engineer biomaterial and might be an ideal replacement biomaterial for ureteral reconstitution.

Objective To evaluate the clinical effectiveness of laparoscopic (LDP) versus open distal pancreatectomy (ODP) using meta-analysis.Methods Comprehensive literature search was conducted on articles only in English published from 2006 to 2012 on MEDLINE,EMbase,Cochrane Central Registry of Controlled Trials to compare LDP with ODP for Pancreatic disease.Data were extracted and evaluated by two reviewers independently.The quality of the included trials was evaluated.Meta-analyses were conducted using the Cochrane Collaboration's RevMan 5.1 software.Results Fourteen controlled clinical trials (n=1417) were included.The LDP group was significantly longer than the ODP group in operation time,and was significantly larger in the number of patients with spleen preservation [(MD-273.10,95％ CI-354.39-191.081,P＜0.01),(OR 2.42,95％ CI 1.78-3.30,P＜0.01) respectively].The LDP group was significantly less than the ODP group in intraoperative blood loss,time to oral intake,and length of hospital stay [(MD-273.10,95％ CI -354.39-191.81,P＜0.01),(MD-1.78,95％ CI-2.36-1.20,P＜0.01),(MD-3.15,95％ CI-3.97-2.33,P＜0.01) respectively].There were no significant differences in blood transfusion,pancreatic fistula rate,and mortality between the two groups.Conclusions LDP is feasible and safe in treating pancreatic disease.When compared with ODP,LDP has the advantages of having less intraoperative blood loss,quicker recovery and more patients with spleen preservation.

Objective To investigate the renal level of 1o-hydroxylase and the change of serum calcium in rats with severe acute pancreatitis,and their correlation.Methods Eighty male Wistar rats were randomly (random number) divided into two groups:sham operation group (SO group),severe acute pancreatitis group (SAP group),and each group was further randomly divided into 1 h,3 h,6 h,and 12 h subgroups (n =10).Severe acute pancreatitis model was made by retrograde infusion with 5％ sodium taurocholate solution into the biliopancreatic duct,rats were sacrificed at 1 h,3 h,6 h,and 12 h separately after modeling.The levels of serum amylase,serum calcium,serum urea nitrogen,serum creatinine,serum 1,25-dihydroxy vitamin D3 were measured,and the level of lα-hydroxylase protein in the kidney was determined with immunohistochemistry and Western blotting.The histopathologic changes of kidney tissue were observed under light microscope and the changes of the proximal tubular epithelial cell were observed under electron microscope.Results Compared with SO group,the levels of serum amylase,serum urea nitrogen,and serum creatinine were higher in SAP group,but the levels of serum calcium and 1,25-dihydroxy vitamin D3 decreased at 3,6,and 12 h,and the renal level of 1 α-hydroxylase also decreased at 3,6,and 12 h after modeling.In SAP group,the levels of serum calcium,serum 1,25-dihydroxy vitamin D3 and the renal level of 1 α-hydroxylase gradually decreased,and the renal level of 1 α-hydroxylase and the level of serum 1,25-dihydroxy vitamin D3 had positive correlation at 3 h,6 h,and 12 h (r =0.93,P ＜0.01; r=0.951,P ＜0.01; r =0.92,P ＜0.01; r =0.878,P ＜0.01),and the renal level of 1α-hydroxylase and the level of serum calcium had positive correlation at 3 h,6 h,and 12 h (r =0.975,P ＜0.01; r=0.946,P＜0.01; r=0.747,P＜0.01).Conclusions Intheearly course of SAP,the lowered activity of 1 α-hydroxylase may play an important role in the development of hypocalcemia.

Objective To study the effect and histocompatibility of urethral extracellular matrix in repair of traumatic urethra defects in rabbits. Methods Model of traumatic urethral defects was made by resecting 1.0-1.5 cm segment of the urethra in 20 rabbits. Then, the defects were repaired by a tube of extracellular matrix of the same length. The dynamic changes of CD4+, CD8+ T cell and CD4+/CD8+ in peripheral blood were detected by flow cytometry at 1,2, 3 and 4 weeks after operation. In the meantime, the immunity response of rabbits was evaluated by lymphocyte transformation test. The repaired segments stained with hematoxylin-eosin (HE) and Van Giesen were studied by histologic and pathologic observations at 10 days, 3, 6 and 24 weeks postoperatively. The urodynamics, urethroscopy and urethrography were performed at 24 weeks postoperatively. Results There was no significant difference in aspects of stimulative index of lymphocyte transformation, T lymphocyte subsets CD4+, CD8+ T cell and CD4+/CD8+ between experimental group and negative control group. Urothelium covered the whole surface of the matrix tube three weeks after operation. The smooth muscle cells increased nearly to normal urethral wall at 24 weeks. Urethrosoopy and urethrography showed glossy matrix tube. There was no statistical difference upon urodynamies between experimental group and control group. Conclusion The urethral extracellular matrix has good histocompatibility and may be a safe and effective material for repairing urethra defects.

Objective To evaluate the protection of 3-aminobenzamide (3-AB),an inhibitor of Poly (ADP-ribose) polymerase (PARP),on severe acute pancreatitis associated adenohypophysis injury in rats.Method Forty Wistar rats were randomly divided into 4 groups:sham operation group (SO group,n=8),SAP group (n=12),3-AB pretreatment group (n =12),drug control group (n =8).The bilepancreatic duct was cannulated through the duodenum and SAP model was induced by a standardized pressure-controlled retrograde infusion of 5％ sodium taurocholate (0.1 ml/100 g) into the bile-pancreatic duct.In 3-AB group,3-AB (20 mg/kg) was administered via femoral vein 30 min prior to the operation;other procedures were identical to SAP group.In SO group,pancreas was flipped several times only.In drug control group,3-AB (20 mg/kg) was administered via femoral vein 30 min prior to the operation.Serum amylase,lipase were measured.Pancreas and pituitary tissue were taken for pathological examination under light microscope.PARP and NF-κB antibodies for adenohypophysis immunohistochemical stains.Adenohypophysis cell was observed under electronic microscope.Result Serum amylase,lipase and pancreas pathological scores were significantly higher in 3-AB group compared with SO group (P ＜ 0.05),but lower than that in SAP group (P ＜ 0.05).Adenohypophysis pathological injury was less severe in 3-AB group.Expressions of PARP and NF-κB in adenohypophysis cells were significantly higher in 3-AB group compared with SO group,but lower than that in SAP group (P ＜ 0.05).Ultrastructural change of thyrotroph cell was relieved in 3-AB group.No significant difference was observed between SO group and drug control group in PARP and NF-κB expression nor adenohypophysis pathological injury.Conclusions 3-AB exerts the protective effect against acute pancreatitis associated adenohypophysis injury by inhibition of PARP and NF-κB.

Objective To investigate the changes of poly-ADP-ribose polymerase (PARP) and NF kappa B (NF-κB) in adenohypophysis in rat model of severe acute pancreatitis (SAP),and their role in the mechanism of adenohypophysis injury in SAP.Methods Forty Wistar rats were randomly (random number) divided into 5 groups:the sham operation group (SO group,n =8),SAP 1 h,3 h,6 h and 12 h groups (n =8 in each group).SAP model was induced by retrograde injection of 5％ sodium taurocholate into the biliopancreatic duct.Serum levels of amylase,lipase and ascites were measured.After sacrifice of experiment rats,pancreas and adenohypophysis tissues were taken for pathological examination under light microscope.Adenohypophysis cells were observed under electronic microscopy as well.PARP and NF-κB expressions in adenohypophysis cell was studied by using immunohistochemisty assay.Results After modelling,serum levels of amylase,lipase and ascites in SAP group increased gradually,which were higher than those in SO group (P ＜ 0.05).Adenohypophysis cell swelling and partial necrosis were observed under light microscope.As the time prolonged,their nuclei became dark and pyknotic more and more,and the endoplasmic reticulum and mitochondrial swelling in adenohypophysis cells were observed under electronic microscopy.The expressions of PARP and NF-κB in SAP group increased gradually,which were higher than those in SO group.Conclusions Significant pathological and ultrastructural injuries were observed in adenohypophysis cells in severe acute pancreatitis.These changes might correlate with PARP and NF-κB signaling pathway.

Objective To explore the effects of rosiglitazone (ROSI),a PPAR-γ ligand,on hyperlipidemia with severe acute pancreatitis (SAP) in the rat model induced by sodium taurocholate injected into intra-bile-pancreatic duct and explore their underlying mechanism.Methods A total of 120 male SD rats were randomly divided into two groups,and eighty rats were fed with high fat diet for two weeks to induce experimental hyperlipemia and the rest received normal diet.The rats fed with normal diet were divided into two groups:sham operation group (SO group,n =20) and SAP group (n =20).The hyperlipidemic rats were randomly divided into four groups:sham operation hyperlipidemia rats group (HL group,n =20),hyperlipidemia with acute pancreatitis group (HAP group,n =20),ROSI prevention group (HR group,n =20) and antagonist group (HRI group,n =20).Rats of SAP group and HAP group were induced by a retrograde infusion of 5％ sodium tauroholate into bile-pancreatic duct,whereas the rats in SO group and HL group were induced by saline instead; rats in HR group were administered ROSI (10 mg/kg) intra-peritoneally 1 hour prior to sodium taurocholate; rats in HRI group were administered GW9662 (0.3 mg/kg) intraperitoneally 30 min prior to ROSI.Rats from each group were sacrificed by exsanguination 12 h after the induction of pancreatitis.Blood samples were taken from all animals to measure serum amylase (AMY),total cholesterol (TC),triglycerides (TG).The severity of pancreatitis was evaluated by histological score of pancreatic injury.The level of nuclear factor (NF)-KB p65 protein in pancreas was measured by immunohistochemistry.The levels of intercellular adhesion molecule (ICAM-1) protein and tumor necrosisfactor-α (TNF-α) protein were detected by using Western blot analysis.Results The serum levels of TC and TG in HL group and HAP group were significantly higher than those in SO group and SAP group (10.86 ± 1.47,10.42±0.95vs.1.72±0.13; 1.24±0.28,1.36±0.13 vs.0.61 ±0.12,0.54±0.08; all P＜ O.05).Compared with SAP group,the levels of serum AMY,the pancreas pathological score,the levels of NF-KB p65,ICAM-1 and TNF-α in pancreas in the HAP group were significantly higher in HAP group (P ＜ 0.05).Compared with the HAP group and HRI group,HR group significantly decreased the levels of serum AMY,TC and TG; pancreas pathological score; the levels of NF-KB p65,ICAM-1 and TNF-α in pancreas significantly decreased in HR group (2006.9 ± 331.9 vs.6501.9 ± 3771.0,5892.2 ± 474.3 ; 4.36 ± 0.99 vs.10.42 ±0.95,11.08 ± 1.05; 0.58 ±0.12 vs.1.36 ±0.13,1.58 ±0.12; all P ＜0.05),but there were no statistically significant differences in those biomarkers between HAP group and HRI (P ＞ 0.05).Conclusions Our study demonstrated that hyperlipidemia aggravated the severity of sodium taurocholateinduced severe acute pancreatitis and ROSI exerted anti-hyperlipidemic effect and anti-inflanmatory effect against hyperlipidemia rats with sodium taurocholate-induced severe acute pancreatitis.

Objective To investigate the effects and mechanisms of poly adenosine diphosphate (ADP)-ribose polymerase inhibitor 3-aminobenzamide (3-AB) on kidney injury in rates with severe acute pancreatitis (SAP).Methods Fifty-six male Wistar rats were divided into the sham operation (SO) group,SAP (3,6,12 hours) groups,and 3-AB + SAP (3,6,12 hours) groups,and there were 8 rats in each group.SAP model was established by retrograde injection of 5％ sodium taurocholate into the biliopancreatic duct.Rats in the 3-AB + SAP group were infused with 3-AB (20 μg/g) via femoral vein 30 minutes before SAP model establishment.The serum amylase,kidney function and renal myeloperoxidase (MPO) were determined,and pathological scores of pancreatic and renal tissues were evaluated under light microscope.Renal poly ADP-ribose formation,intercellular adhesion molecules-1 (ICAM-1) and P-selectin expression were detected by the Western blot.All data were analyzed using the analysis of variance or t test.Renal injury grading was analyzed using the Kruskal-Wallis nonparametric test.Results The levels of serum amylase of SAP 3,6,12 groups were (3806 ± 229)U/L,(4898 ± 295) U/L and (5726 ± 372) U/L,which were significantly higher than (2785 ± 160) U/L,(3241 ± 198) U/L and (3953 ± 249) U/L of the 3-AB + SAP groups (t =3.652,4.672,4.407,P ＜ 0.05).The levels of blood urea nitrogen were (11.6 ± 0.8) mmol/L,(19.3 ± 1.3) mmol/L and (29.6 ± 2.1) mmol/L,which were higher than (7.5 ± 0.5) mmol/L,(10.5 ± 0.7) mmol/L and (21.6 ± 1.5) mmol/L of the 3-AB + SAP groups.There were significant differences in the levels of blood urea nitrogen between the SAP group and the 3-AB + SAP group at the 6 and 12 hours (t =3.836,6.849,P ＜0.05).The levels of creatinine of the SAP 3,6,12 hours groups were (48.7 ±3.1) μmol/L,(58.3 ±3.7) μmol/L and (75.9 ±5.4) μmol/L,which were higher than (40.7 ±2.6)μmol/L,(43.2 ± 2.6) μmol/L and (53.4 ± 3.2) μmol/L of the 3-AB + SAP groups.There were significant differences in the levels of creatinine between the SAP group and the 3-AB + SAP group at the 6 and 12 hours (t =3.279,3.073,P ＜ 0.05).The renal MPO activity of the SAP 3,6,12 hours groups were (0.69 ± 0.06) U/g,(1.07 ± 0.09)U/g and (1.42 ±0.13)U/g,which were higher than (0.57 ±0.05)U/g,(0.75 ±0.06)U/g and (0.89 ± 0.07) U/g of the 3-AB + SAP groups.There were significant differences in the renal MPO activity between the SAP group and the 3-AB + SAP group at the 6 and 12 hours (t =3.066,4.012,P ＜ 0.05).The pancreatic pathological scores of the SAP 3,6 and 12 hours group were 6.50 ± 0.53,9.06 ± 0.66 and 11.75 ± 0.89,which were significantly higher than 4.25 ± 0.31,6.06 ± 0.51 and 7.57 ± 0.59 of the 3-AB + SAP group (t =3.631,3.598,5.147,P ＜ 0.05).The structure of the kidney was normal in the SO group.Congestive changes were observed in glomerulus of kidney,the renal tubular epithelial cell was necrosed,and luminal narrowing or occlusion,hemorrhage in the intercellular substance and inflammatory cell infiltration were observed in the SAP 12 hours group.The pathological changes of the 3-AB + SAP 12 hours group were significantly slighter (P ＜ 0.05).The relative expressions of poly ADP-ribose,ICAM-1 and P-selectin of the SO group were 1.00 ±0.21,1.00 ±0.18,1.00 ± 0.16,which were significantly lower than 3.83 ± 0.63,5.42 ± 0.83,3.71 ± 0.48 of the SAP 12 hours group (t =6.955,23.107,10.352,P ＜ 0.05).The relative expressions of poly-ADP-ribose,ICAM-1 and P-selectin of the 3-AB + SAP 12 hours group were 1.94 ± 0.36,2.35 ± 0.35,2.11 ± 0.29,which were significantly lower than SAP 12 hours group (t =3.977,12.115,5.012,P ＜ 0.05).Conclusions Poly ADP-ribose polymerase inhibitor 3-AB protects kidney from injury in the experimental SAP rats.Poly ADP-ribose polymerase inhibitor 3-AB functions by suppressing the ICAM-1 and P-selectin expression and reducing neutrophil infiltration.