Aim To investigate the interaction between regulatory proteins related to the activation of eNOS and CYP4A/20-HETE.Methods The endothelia of pulmonary artery of new born bovine were cultured and divided into four groups: controlling group: ethanol solvent as vehicle (the volume is the same with 1?10~ -6 mol?L~ -1 20-HETE) was added to culture dish and incubated for 10 min; 20-HETE 5 min treatment group:1?10~ -6 mol?L~ -1 20-HETE was added to culture dish and incubated for 5 min; 20-HETE 10 min treatment group:1?10~ -6 mol?L~ -1 20-HETE was added to culture dish and incubated for 10 min; VEGF 10 min treatment group: 1?10~ -8 mol?L~ -1 VEGF was added to culture dish and incubated for 10 min. CYP4A,eNOS and proteins related to the activation of eNOS were measured by Immunoprecipitating and immunobloting.Results Expressions of eNOS and phospho-eNOS(Ser1177)in PAECs were upregulated after the cells were treated with 20-HETE, and similar phenomena were observed when 20-HETE was replaced by VEGF; 20-HETE increased the binding of Hsp、Akt and eNOS.Conclusions ①20-HETE phosphorylates eNOS at Ser1177 site which activates the eNOS to catalyze L-aginine to produce nitric oxide.②eNOS binds with Hsp90 and Akt in endothelial cell of pulmonary artery and 20-HETE increase the binding. ③eNOS binds with CYP4A in endothelial cell of pulmonary artery.

Objective To observe the ultrastructural characteristics of human retinal progenitor cells cultured in vitro. Methods Six 5-month-old human fetuses(12 eyes)without eye diseases were selected. Retinal progenitor cells from the retina of one eye of each fetus were cultured in vitro,and observed by transmission electronic microscopy(TEM); while those from the other eye were directly observed by TEM. Results Abundant heterochromatin were found in the karyon of 5-month embryonic retinal neuroepithelial cells,and the figure of the karyons was irregular.A few scattered initial cells were seen in retinal neuroepithelial layer with large karyon,smooth surface,abundant euchromatin,and distinct nucleolus.The human retinal progenitor cells cultured in vitro had the same ultrastructural characteristics as the initial cells:with huge karyon which almost occupied the whole cell,little cytoplasm,distint nucleolus,abundant euchromatin,and little heterochromatin.The cells clung to each other in the neural globoid cell mass.The size of the outer cells was large,and karyokinesis could be found. Conclusion The cultured human retinal progenitor cells are provided with the same ultrastructure characteristics as the initial cells.

Objective To explore the relationship between ischemia/reperfusion (IR) and the expression ET-3 and GFAP. Methods Thirty-six adult male mice were randomly divided into three groups: ⑴ ischemia/reperfusion group(IR,n=24), bilateral common carotid arteries(BCCA) of mice were ligated for 7 minutes and reperfused for 1day,3day,5day and 10day; ⑵ a sham operation group (SO, n=6); ⑶ normal control group(NG, n=6). The ET-3 and GFAP expressions in the layers Ⅲ-Ⅵ of frontal and parietal lobes of mice in the three groups was detected by immunohistochemisty. Results The expressions of ET-3 and GFAP in the layers Ⅲ-Ⅵ of frontal and parietal lobes significantly increased in I/R group(P

Objective To observe the long-term effect of suramin on the inhibition of proliferation of human retinal pigment epithelial (RPE) cells in vitro. Methods RPE cells grown in 9 pieces of 96-well plate (12 wells each plate) were divided into experimental and control group, with 6 wells in each group. The concentration of 0.1 ml RPE cells in each well is 5?104 cells/ml. After the change of the medium, RPE cells were treated with suramin (250 ?g/ml) in experimental group while treated with nothing in the control group. The medium of the 2 groups were changed to the normal medium after 4 days. At the 1~st , 2~nd , and 4~th day after the addition of suramin and at the 1~st , 2~nd , 3~rd , 5~th , 6~th , 7~th , 9~th , 11~th and 13~th day after removing suramin, 1 plate was randomly selected to stop culturing, and the proliferation of RPE cells were detected by methyl thiazolyl tetrazolium (MTT) assay. Results Under reversed microscope, RPE cells in control group were fused completely at the 7~th day after inoculation. The extracellular space of RPE cells in experimental groups was larger than that in the control group, and remained unfused at the 13~th day after inoculation. The inhibitory rate of proliferation of RPE cells at the first day after treated with suramin was 14.85% and increased to the highest 25.79% at the 4~th day. The first day after the suramin-containing media was removed, the inhibitory rate decreased to 12.35%, and then raised gradually to over 20% at the 3~rd to 5~th day. Finally, the rate drop to 14.71%. Conclusion Suramin has the long-term effect on the inhibition of RPE cells induced by serum, especially the inhibitive effect after the remove of suramin, which indicates the specific double-peak inhibition during the whole process.

Objective To investigate the prognostic factots by analyzing clinicopathologlcal characteristics of 972 cases of colonic carcinoma. Methods Clinical and pathological data of 972 patients with colonic carcinoma treated surgically were studied by univariate and multivariate analysis,survival rate was calculated by life-table method and compared by Log-rank test. Results The univariate analysis indicated that the predictors of survival were age,perioperative blood transfusion, perioperative carcinoembryonic antigen(CEA)level,tumor gross type,depth of tumor invasion,lymphatic invasion,liver metastasis,extrahepatic metastasis,local recurrence、peritoneal seeding,pathological type,TNM stage and type of lymph node clearance.The multivariate analysis indicated that the independent predictors of suivival were age,periopemtive CEA level,tumor gross type,lymphatic invasion,liver metastasm,extrahepatic metastasis,local recurrence、peritioneal seeding,pathological type,type of lymph node clearance and TNM stage. Confclusions Lymphatic invasion is the most important independent prognostic factor of patients with colonic carcinoma treated surgically.

It is usually a challenge to repair the abdominal wall defect to general surgeons. With developing of materials science, the use of mesh provides a novel method of primary closure of abdominal wall defects in this set-ting. Recently , biological mesh has been reported to reconstruct abdominal wall successfully. This review is to in-troduce the recent status and progress on its biological characteristics,animal experiments and clinical Study.

OBJECTIVE:To observe analgesic effect and safety of ketorolac tromethamine combined with butorphanol tartrate in the treatment of acute pain after fracture surgery. METHODS:76 acute pain patients after fracture surgery were selected and ran-domly divided into control group and observation group,with 38 cases in each group. Control group was given Ketorolac trometh-amine injection 30 mg,ivgtt,and then 2 ml/h,0.5 mg/kg,ivgtt;observation group was additionally given butorphanol tartrate 10 mg,ivgtt,on the basis of control group. Pain degree was evaluated with VAS before and 10 min,1,2,4 and 6 h after treatment, and the occurrence of ADR was observed in 2 groups. RESULTS:10 min,1,2,4 and 6 h after treatment,VAS score of 2 groups were significantly lower than before,with statistical significance(P<0.05);6 h after treatment,VAS score of observation group was significantly higher than that of control group,with statistical significance (P<0.05);there was no statistical significance in VAS score between 2 groups 10 min,1,2 and 4 h after treatment (P>0.05). The incidence of ADR in observation group (5.26%)was significantly lower than in control group(21.05%),with statistical significance(P<0.05). CONCLUSIONS:Com-pared with ketorolac tromethamine alone,ketorolac tromethamine combined with butorphanol tartrate shows shorter analgesia dura-tion,similar therapeutic efficacy,and lower incidence of ADR.

Objective To establish diagnosis model and explore related metabolic pathways by analyzing the serum metabolic profile of patients with primary nephrotic syndrome (PNS) through metabolomics.Methods Thirty PNS patients hospitalized in Huai'an First People's Hospital between December 2010 and April 2012 were enrolled.High performance liquid chromatography-mass spectrometry (LC-MS) was employed to detect metabolites in the serum of 30 PNS patients and 30 healthy controls.Metabolic fingerprint profiling and multivariate pattern recognition analysis were combined to establish disease-specific metabolic diagnosis model,and metabolic pathway analysis was performed.Results PNS group and control group could be well separated by principal component analysis (PCA) model as well as partial least-squares discriminant analysis (PLS-DA) model with Q2 of 0.300.There was well interpretation in PLA-DA model (R2X=0.581,R2Y=0.452).Compared with healthy controls,PNS patients had decreased cholestane 3,7,12,15 alcohol,acyl glycerine,phytosphingosine and tryptophan,and increased sphingomyelin,arginine and glutamic acid (all VIP ＞ 1,P ＜ 0.05).The metabolic disorders pathways of PNS patients included sphingolipid metabolism,arginine and proline metabolism,linoleic acid metabolism and pyrimidine metabolism (all impact ＞0.10 and P ＜ 0.05).Conclusions Metabolomics combined with multivariate pattern recognition analysis may be a new tool for diagnosis and monitoring of PNS.

AIM: To explore the effects of hydrogen sulfide (H_2S) on proliferation of vascular smooth muscle cells (VSMC) stimulated by endothelin (ET-1, 10 -7 mol/L ) and mitrogen-activated protein kinase (MAPK) activity in VSMCs. METHODS: Cultured VSMCs were divided into six groups: (1) control group, (2) serum group, (3) endothelin group, (4) NaHS groups, (5) serum+NaHS group, and (6) endothelin+NaHS group. VSMC proliferation was measured by [ 3H]-TdR incorporation and MAPK activity in VSMC was determined by radioactivity assay. RESULTS: ET-1 increased VSMC [ 3H]-TdR incorporation by 2.39 times ( P

Objective To investigate the effect of platelet-rich plasma (PRP) in anterior cruciate ligament (ACL) reconstruction.Methods From January 2010 to January 2013,40 patients with ACL ruptures who underwent arthroscopic ACL reconstruction with gracilis and semitendinosus tendon were randomly divided into two groups:PRP group and normal saline group.20 patients received graft soaked with PRP and hemocoagulase while 20 patients received graft soaked with normal saline and hemocoagulase.All patients were followed up in 1,3 and 12 months.Evaluation consisted of postoperative drainage volume,inflammatory reaction,grade of wound healed,anterior drawer test,Lachman test,pivot shift,Lysholm knee score and KNEELAX3.Results The average follow-up period was 18 months.Postoperative drainage volume was 142±24 ml in PRP group and 324±22 ml in saline group.The difference was statistically significant.At 4 days after the operation,no inflammatory reaction was observed in 18 cases of PRP group and in 16 cases of saline group,mid inflammatory reaction in 1 case of PRP group and 2 cases of sa line group,and moderate inflammatory reaction in 1 case of PRP group and 2 cases of saline group.Wound healed by first intention in 20 patients of PRP group and in 19 patients of saline group.The preoperative results of anterior drawer test,Lachman test and pivot shift were positive,while postoperative results were negative in both two groups.In PRP group,the preoperative and postoperative Lysholm knee scores of patients in 12 months were 39.8±8.9 and 92.1±2.7 points respectively.In saline group,the preoperative and postoperative Lysholm knee scores of patients in 12 months were 38.7±9.8 and 89.9±4.1points respectively.The differences were not statistically significant.KNEELAX3 measuring results:in PRP group,preoperative measurement was 9.4±1.2 mm in average,while measurement in 12 months postoperatively was 1.2±1.1 mm.In saline group,preoperative measurement was 9.6±1.3 mm,while measurement in 12 months postoperatively was 2.2±1.2 mm.The differences were statistically significant.Conclusion Using graft soaked with PRP in ACL reconstruction could reduce postoperative drainage volume,accelerate the healing of tendon-bone interface in the bone tunnel and the recovery of knee joint function.