Objective To develop a method of adult porcine pancreatic islet isolation.Methods The tails of adult porcine pancreas were perfused through the pancreatic duct with 0.1% cold collagenase(type Ⅺ) and incubated at 38.5 ℃.The digested tissue was dispersed in 4 ℃ Hanks balanced salt solution(HBSS).The tissue suspensions were filtered through a 600 ?m mesh.The residual tissue was resuspended in cold HBSS,and put in the Ricordi’s chamber and shaken for 5 minutes,then filtered again.The isolated islets were divided into three groups: control group( n =14),Pefabloc(trypsin inhibitor, n =8) group and FOY(trypsin inhibitor, n =5) group.The collagenase solution of the Pefabloc and FOY group was supplemented with 1.0 mmol/L Pefabloc and FOY respectively. Results The islet yields of the Pefabloc group and FOY group 〔(11 848?3 530) islet/g pancreas and (14 496?3 693) islet/g pancreas〕 were significantly higher than that of the control group 〔(8 505?3 349) islet/g pancreas〕, P

Objective To construct and appraise a series of shRNA eukaryotic expression plasmid which contains vary spots of human SKP2 gene.Methods Three pairs of hairpin-like oligonucleotide sequences specific for human SKP2 gene were designed and synthesized. They were cloned into the eukaryotic expression pSIREN-RetroQ plasmid. The recombinated plasmids were confirmed by PCR and sequencing.Results The shRNA sequences were constructed successfully, and could be inserted into the eukaryotic expression vector pSIREN-RetroQ.Conclusions Three shRNA transcripting plasmids pSIREN-RetroQ are successfully constructed, and form a basis for research in cancer gene target therapy against SKP2 gene.

Objective To investigate the effect of TLCK(N-alpha-tosyl-L-lysyl-chloromethylketone) on acute hemorrhagic necrotizing pancreatitis(AHNP) associated with lung injury.Methods Seventy SD rats were randomized into seven groups(n=10 in each group): group1,normal control group(rats were treated with 0.9% sodium chloride i.v.);group 2,AHNP control group(AHNP rats were treated with 0.9% sodium chloride i.v.);group 3,AHNP rats were treated with 5?g/kg TLCK;group 4,AHNP rats were treated with 10?g/kg TLCK;group5,AHNP rats were treated with 20 ?g/kg TLCK;Group3,4,5were treated with TLCK i.v.immediately after the establishment of SAP rat model;group 6,AHNP rats were treated with TLCK i.v.30 minutes before the establishment of AHNP model;group 7,AHNP rats were treated with TLCK i.v.30 minutes after the establishment of AHNP rat model;Group 6,7 were treated with 10 ?g/kg TLCK.Group 1-7 were used to select the best dosage and administration time of TLCK by observing the survival rate for seven days.Then,according to the result of selecting,the SD rats were divided into three groups(n=6 in each group): group N,Sham operated control group(0.9% sodium chloride was reversely injected into SD rat′s pancreas and rats were treated with 0.9% sodium chloride i.v.);group P: AHNP group;group T: TLCK treatment group(AHNP rats were treated with 10 ?g/kg TLCK immediately after the establishment of AHNP rat model).The rats of the last three groups were killed 6 hours after operation.Alveolar macrophages(AMs) were harvested by bronchoalveolar lavage.The protein content of bronchoalveolar lavage fluids(BALF),the myeloperoxidase(MPO) of lung tissue,and tumor necrosis factor ?(TNF?) secreted by AMs were determined.The expression of NF-?B in AM was checked as well as lung histology.Results The survival rate of group 1-7 was100%,0%,70%,100%,80%,0%,and 90% respectively.The level of MPO of lung tissue and protein content of BALF ingroup N were lower than that,in groups P and T(P

Objective To explore the effect of nitric oxide(NO) on pulmonary tumor necrosis factor?(TNF?) in murine acute necrotizing pancreatitis(ANP). Method Ninety-six SD rats were randomized into four groups:normal control group, ANP group, L-Arginine(L-Arg) pretreatment group and L-NAME pretreatment group (n=24 for each group). The protein content of bronchoalveolar lavage fluids(BALF), the myeloperoxidase(MPO) of lung tissue and TNF? produced by alveolar macrophages were evaluated. The expression of TNF? mRNA was measured. ResultMPO and protein of BALF reached a peak level at 12nd hr (10.78?0.58U/g for MPO and 2011?106?g/ml for protein content respectively).TNF? peaked on the sixth hour(1624?149)pg/ml. The expression of AM TNF?mRNA also peaked on the sixth hour (1.127?0.069) along with an increase of TNF? mRNA. A similar tendency was seen in L-Arg and L-NAME pretreatment groups, with changes being statistically different in the three groups when compared with that of normal control group(P

Objective To investigate the relationships between the expression of cyclooxygenase-2 (COX-2) , vascular endothelial growth factor (VEGF) and the degree of vascularization, clinicopathological feature, survival time of the patients with gallbladder carcinoma. Methods Routine paraffin-embeded sections of gallbladder carcinoma tissues in 64 cases were evaluated for COX-2, VEGF expression and MV by the streptavidin-peroxidase complex immunohistochemical method. Results COX-2, VEGF immunoreactivity were observed in 72%,and 55% cases, respectively. The average MVC was (57?14)/ HP. The status of MVC was closely correlated with Nevin staging, tumor differentiation and lymph node metastasis (P well differentiated,P0.05). There was a positive correlation between COX-2 expression and clinical stages. The positive rate of COX-2 was higher in cases of Nevin stages S4-S5 (82%) with lymph node metastasis than in those of Nevin stages S1 -S3 (50%) and without metastasis (P0.05 ). The expression of VEGF and COX-2 was significantly correlated with that of MVC (t=5.424, P

Objective To discuss the clinical features,diagnosis and operative treatment in Crohn disease(CD)characterized by acute abdomen.Method The clinical data of 26 CD cases testified by pathology after emergency operations from April 1997 to February 2009 were retrospectively analyzed.Results In 26 cases of acute abdomen,17 cases characterized by mechanical obstruction,6 cases characterized by peritonitis,3 cases characterized by hemorrhage of digestive tract.Eight cases(30.8%)were diagnosed before operation,and 13 cases(50.0%)were diagnosed during operation,totally 80.8%(21/26).All patients received surgical operations as following:resection and anastomosis in 21 eases,resection and fistulation in 4 cases,ileotransverso-stomy in 1 case.Two cases died after operation,and others discharged including 1 case received second operation within 5 days.Conclusion It is difficult to be diagnosed before operation when CD characterized by acute abdomen,and appropriate emergency operations are needed according to the particular cases.

Objective To investigate the mechanism of endothelin (ET) in acute biliary pancretitis (ABP)with renal impairment, and examine the improvement of renal function by puerarin. Methods Thirty-two patients of ABP with renal impairment were selected and were randomly divided into group A and group B. Group A were given puerarin post-operatively, while group B were given common manegement postoperatively. Meanwhile, 16 ABP patients without renal impairment were selected as group C, and 16 nonABP patients who had common hepatic or biliary diseases were selected as group D. The levels of plsma ETand creatinine clearance rate(Ccr) of renal function in each group were observed pre-operation and 1 week post operation. Results In pre-operation the ET level in group A, B and C were higher than in group D(P ＜0.001), and the ET level in group A,B was higher than in group C(P＜0.001). Post operation after I week the ET level in group A was lower than in group B(P =0.014), while the Ccr level in group A was higher thanin group B ( P = 0.002). Conclusions The function of endothelin is an important mechanism in biliary pancretitis with renal impairment. Puerarin might reduce the ET level and increase the Ccr level in patients of ABP with renal impairment, improve the renal function of such patients.

With the development of systemic therapy for the treatment of breast cancer, the incidence of brain metastases from breast cancer is gradually increasing. Treatment with surgery,stereotactic radiosurgery, and wholebrain radiotherapy (WBRT) are still the first-line therapies for the majority of patients with brain metastases. Recently, chemotherapy, hormone therapy and targeted therapy have been widely used for breast cancer. This review summarizes the efficiency of those strategies for the breast cancer patients with brain metastases.

Objective To discuss the role of activation of nuclear factor ?appa B(NF-?B) in apoptosis of alveolar macrophages(AM) induced by gadolinium chloride(GdCl3) in acute necrotizing pancreatitis(ANP).Methods Thirty sixty adult SD rats were randomized into three groups: normal control group,ANP group and the group treated by GdCl3.ANP was induced by intraductal administration of 5% sodium taurocholate,while the normal control rats received an infusion of normal saline.In GdCl3 treatment group,GdCl3 was injected into dorsal vein of penis right after ANP model was established.AM were harvested by bronchoalveolar lavage six hours after model was established.The generation of TNF-? and IL-1? in bronchoalveolar lavage fluid(BALF),and the level of myeloperoxidase in lung tissue were evaluated.The expression of NF-?B protein in AM was determined by western blot.The apoptosis of AM was detected by agarose gel electrophoresis and flow cytometer.The histological examination of lung tissue was checked.Results The levels of TNF-? and IL-1? in ANP group were significantly higher than the control group and GdCl3 treatment group(P

Objective To assess the clinical value and safety of VenaTech LP permanent inferior vena cava filter (IVCF) for preventing pulmonary embolism (PE) in patients with deep vein thrombosis (DVT). Methods One hundred and seven DVT patients receiving VenaTech LP permanent IVCF implantation from October 2005 to November 2009 were selected. In hospital and following up data were analyzed. Results All the filters were successfully implanted. No symptomatic PE took place during acute stage of thrombosis after filter implantation. No complication took place during following up 1 to 36 months.Conclusion Implantation of VenaTech LP permanent IVCF is safe and effective in the prevention of PE.