Aim To further research into the antithrombotic mechanism of scorpion venom active peptides (SVAP). Methods Human umbilical vein endothelial cells (HUVEC) were cultured with enzyme digestive method. After the cultured HUVC was incubated in conditioned media for 1 hour, the effects of SVAP on the concentration of 6-Keto-PGF 1? and NO of HUVEC were determined with radioactive-immunolygic and nitrate reduction enzyme method respectively. Results As compared with control, SVAP in the doses of 1,5,10, 20 mg?L -1 had the distinctive increase of 54%, 68%,72%,79% of the concentration of 6-Keto-PGF 1? and SVAP in the doses of 10, 20 mg?L -1 had the significantly increased of 27%, 46% of the concentration of NO. Regression anylysis showed that the release levels of PGI 2 and NO in HUVEC induced by SVAP was of positive correlation. Conclusion Antithrombotic mechanism of SVAP is related to the increase of PGI 2 and NO released from HUVEC and synergistic and mediating action between NO and PGI 2.

Objective To assess the technical feasibility and efficacy of the combined application of a flexible,self-expanding neurovascular stent(Neuroform)and Gugliebni detachable coils(GDC)in the management of wide-necked intracranial aneurysms in humans.Methods Sixty-five wide-necked aneurysms which underwent 65 endovascular procedures were performed by using intracranial stent and GDC.There was a total of 30 aneurysms at basilar artery including 16 at the basilar tip,9 at the basilar trunk and 5 at the beginning of the basilar artery.And there were 30 aneurysms located at the posterior communicating artery, and 5 aneurysms located at the vertebral artery.The Neuroform stents were deployed to cover the neck of aneurysms.Another microcatheter was introduced into the aneurysm sac through the stent interstices and then detachable coils were released to embolize the aneurysms.Results The combined procedures were successful in all of the 65 patients with wide-necked aneurysms.The stent could pass smoothly through the intracranial artery and got released.Complete occlusion was achieved in 60 patients and incomplete occlusion in 5 patients.In-stent thrombosis occurred in 2 patients.All patients recovered well.Forty-two patients had followe-up angiography at 3 to 6 months after the procedure.Among them,no filling was found for the 39 aneurysms which were densely packed,and 3 aneurysms had neck remnant.Conclusion The implantation of Neuroform stent as a complimentary device to GDC coiling is easy and safe for embolization of wide-necked intracranial aneurysms.It has great advantage for treatment of wide-necked intracranial aneurysms.

A.niger M1, the initial strain, was treated by UV and a mutant with 30% higher xylanase activity was obtained. Zymogram for detecting xylanase showed there are three different xylanases in the mutant mature culture, while two xylanases in initial strain. After orthogonal experiment, the optimum fermentation conditions of the mutant were obtained as follows: concentration of the major carbon resource 4 %, ratio between bran and corncob 5:5, concentration of glucose 0.1%, concentration of ammonium oxalate as supplemental nitrogen resource 2.0%, the initial pH of liquid medium 5.0, 100mL/250mL flask.

Objective To investigated the effect of procyanidin (PC) on the expression of cysteine proteinase -3 (Caspase -3) in type 2 diabetes mellitus SD rats with focal cerebral ischemia. Methods Following the random principle, 40 healthy Sprague - Dawley (SD) rats were numbered sequentially and randomly divided to normal rats with focal cerebral ischemia group,type 2 diabetes mellitus SD rats with focal cerebral ischemia group,PC low/ middle/ high -dose groups,with 8 rats in each group. The type 2 diabetes mellitus - MCAO model was set up. The doses of PC for low,middle and high - dose groups were 50 mg/ kg,100 mg/ kg,200 mg/ kg. Immunohistochemistry method was used to measure the activity of Caspase - 3. Results Compared with that in the normal rats with focal cerebral ischemia group[(11. 42 ±2. 52)],the expression of Caspase -3 increased in the type 2 diabetes with ischemia group[(15. 00 ± 2. 38)](t = 2. 17,P < 0. 01). Compared with that in the type 2 diabetes with ischemia group,the expression of Caspase - 3 decreased in the PC groups[(9. 38 ± 2. 00),(7. 71 ± 1. 55),(6. 96 ± 1. 57)](t = 2. 86,3. 13,3. 36,all P < 0. 01),whereby the middle and high - dose groups showed more significant decrease (t = 1. 92,2. 03,all P <0. 01) and with no statistically significant difference between the two groups(t = 1. 13,P > 0. 05). Conclusion PC can decrease the expression of Caspase - 3 protein in type 2 diabetes mellitus SD rats with focal cerebral ischemia, finally may inhibit the apoptosis.

Objective To investigate the clonal types of Staphylococcus aureus collected from 2004to 2010 in patients with blood stream infection from a Grade A tertiary care hospital in Shanghai as well as the dynamic changes and to detect the variation in antimicrobial resistance and virulence-gene content in different strain types.Methods A total of 103 nonduplicate S.aureus isolates were collected from 2004 to 2010 from inpatients with S.aureus blood stream infection from Shanghai Huashan hospital.Determination of oxacillin MICs and the type of SCCmec gene were used to screen MRSA.Typing of S.aureus isolates was identified by using multilocus sequence typing(MLST) and S.aureus-specific staphylococcal protein A typing(spa typing),PCR was used to detect the antimicrobial resistance and virulence-gene.Results Sixtysix isolates(64.1%) MRSA were detected in 103 nonduplicate S.aureus isolates,and 35 isolates were MRSA with SCCmec type Ⅱ ,Twenty-nine isolates were MRSA with SCCmec type Ⅱ,two isolates were MRSA with SCCmec type Ⅳ,Thirty-seven isolates(35.9%) were MSSA.Thirty-three MRSA isolates were ST5,Twenty-nine MRSA isolates were ST239,two MRSA isolates were ST59,one MRSA isolates was ST641 and one MRSA isolates was ST6.All of the other clones belonged to MSSA.The percentage of ST5 and ST239 were decreased significantly after 2009(ST5 was decreased from 52.9% to 15.4%; ST239 was decreased from 61.1% to 15.4%),and new clonal types MSSA increased significantly(in 2009,the percentage of ST7 was 41.7%; new clonal types such as ST188 and ST15 were detected in 2010).In 2010,it was shown that 84.6% of MSSA were isolated from S.aureus blood stream infection,nineteen isolates(18.4%) harbored mupA gene and 41 isolates(39.8%) harbored qacA/B gene in 103 nonduplicate S.aureus isolates.It was shown that 70.6% ST239 harbored qacA/B gene.Four isolates of ST398 and 1 isolates of ST9were detected which were originally from animal.There was no significant difference of the virulence gene presence in the same strain types except sasX、lukSF and arcA genes,but there were a lot of genes which were restricted to different genomic background.Conclusions The percentage of ST5 and ST239 were decreased and new clonal types of MSSA were increased significantly in S.aureus blood stream infection,antimicrobial resistance and virulence-gene were restricted to different clonal types.

Objective To investigate the effects of sevoflurane postconditioning on myocardial ischemiareperfusion(I/R)injury in patients undergoing coronary artery bypass grafting(CABG)with cardiopulmonary bypass(CPB).Methods Forty NYHA Ⅰ -Ⅲ patients of both sexes,aged 55-64 yr,with BMI ＜ 30 kg/m2,scheduled for CABG under CPB,were randomly divided into 2 groups(n = 20): control group(group C)and sevoflurane postconditioning group(group S).Anesthesia was induced with midazolam and/or etomidate,fentanyl and rocuronium.Patients were tracheal intubated and mechanically ventilated.Anesthesia was maintained with iv infusion of propefol and intermittent iv injection of fentanyl and pipecuronium.In group S,2% sevoflurane was inhaled continuously for 15 min immediately after aortic unclamping.After anesthesia induction,before CPB,10 min after the end of CPB,at the end of operation,and 6 and 24 h after operation,MAP,HR,CVP,mean pulmonary arterial pressure,pulmonary arterial wedge pressure,CO and S(v)O2 were recorded,and CI,SVI,systemic vascular resistance index and pulmonary vascular resistance index were calculated.Blood samples were taken from central vein before aortic clamping,at 6 h of reperfusion and 24 h after operation for determination of plasma creatine kinase(CK),creatine kinase isoenzyme(CK- M B)and lactate dehydrogenase(LDH)activities and tropenin I(TnI)concentrations.Myocardial tissues were obtained from right auricle before aortic clamping and at the end of CPB for observation of the ultrastructure and the severity of myocardial injury was assessed.Results There was no significant difference in hemodynamics and parameters of cardiac function between the two groups(P ＞ 0.05).Compared with group C,plasma CK-MB and LDH activities at 6 h of reperfusion and plasma CK activity and TnI concentrations at 24 h after operation were significantly decreased and the myocardial injury was significantly reduced after the end of CPB in group S(P ＜ 0.05).Conclusion Sevoflurane postconditioning can protect myocardium against I/R injury induced by CPB in patients undergoing CABG.

OBJECTIVETo determine the expression of Skp2 in different prostate cancer (PCa) cell lines and tissues, and explore its influence on the androgen receptor (AR) signaling pathway and development of castration-resistant prostate cancer (CRPC).

METHODSThe expression levels of Skp2 and AR in different PCa cell lines were detected by Western blot. After knockdown of Skp2 in the C4-2 and 22RV1 cells transfected with shRNA, the expressions of AR and P27 were determined and the activity of ARR3-Luc measured by dual-luciferase reporter gene assay following treatment with dihydrotestosterone (DHT). The expressions of AR and Skp2 in human naïve PCa or CRPC specimens were detected by immunohistochemical staining followed by analysis of their differences and correlation.

RESULTSThe Skp2 protein expression level was significantly higher in the C4-2 or 22RV1 cells than in the LNCaP cells. DHT treatment increased the expression of Skp2 in the C4-2 cells, but knock-down of Skp2 significantly up-regulated the expression of the well-known downstream protein P27 and down-regulated that of AR. Consistently, DHT treatment increased the activity of ARR3-Luc, while knockdown of Skp2 remarkably decreased it in the C4-2 and 22RV1 cells (P < 0.05). In addition, significantly higher expressions of Skp2 and AR were observed in the CRPC than in the naïve specimens (P < 0.05), with a positive correlation between the two proteins (r = 0.658 1, P < 0.05).

CONCLUSIONSkp2 can enhance the expression and transcription activity of the AR protein in CRPC cells or tissues and is promising to be a critical molecular therapeutic target.

Androgens ; pharmacology ; Cell Line, Tumor ; Dihydrotestosterone ; pharmacology ; Disease Progression ; Gene Knockdown Techniques ; Humans ; Male ; Neoplasm Proteins ; genetics ; metabolism ; Prostatic Neoplasms, Castration-Resistant ; metabolism ; Receptors, Androgen ; genetics ; metabolism ; S-Phase Kinase-Associated Proteins ; physiology ; Transcriptional Activation ; Up-Regulation

ObjectiveTo investigate the distribution and effect on antibiotic resistance of the novel cell wall anchored protein-encoding gene sasX.MethodsA total of 300 S.aureus isolates were randomly collected from inpatients with S.aureusinfection in ShanghaiHuashanhospitalin 2004, 2007and 2010.Meanwhile,170 S.aureus isolates from the nasal swabs of healthy people were collected as part of a population-based community prevalence study.Typing of S.aureus isolates were identified by using multilocus sequence typing (MLST) and S.aureus-specific staphylococcal protein A typing (spa typing).Determination of oxacillin MICs was used to screen MRSA.PCR and sequencing were used to analyze sasX gene.The effect on antibiotic resistance of sasX gene was detect by disc agar diffusion drug sensitive test.ResultsThe major clonal types of the 300 S.aureus isolates collected from inpatients with S.aureus infection were ST239 ( 110,36.7％) and ST5 (122,40.7％).From 2004 to 2010,the percentage of isolates from inpatients with S.aureus infection was increased from 17％ to 39％,but sasX was only found in 0.59％ of the S.aureus isolates from the nasal swabs of healthy people.The percentage of sasX positive was increased from 47.2％ to 83.8％ in ST239.The percentage of sasX positive MRSA was increased from 26.4％ to 50.8％,but the percentage of sasX positive MSSA was about 10％.Antibiotic resistance of sasX positive strains were higher than that of sasX negative strains.Conclusions SasX gene is mainly detected in nosocomial pathogenic S.aureus and it is a possible virulence factor of S.aureus in hosptal setting.The presence of sasX gene is related to antibiotic resistance.For better understanding the real function of this novel gene,further studies such as expression of the encoded protein should be carried out.

Objective To determine whether the novel surface-anchored protein SasX promotes aggregation of S.aureus and adherence of S.aureus to human nasal epithelial cells.Methods MRSA ST-239 HS770 sasX gene mutant ( HS770 △sasX) and complement [ HS770 △sasX (pRBsasX) ] were gotten by gene knock-out and complement methods.The aggregation ability of S.aureus was observed through microscope.By adherence assay which was used for detection of the adherence ability of wild type and mutant to human nasal epithelial cells and blocking experiments which detected the ability of the purified recombinant SasX protein in blocking the adherence of S.aureus to human nasal epithelial cells,we investigated the influence SasX on colonization of S.aureus.Results Compared to wild type,HS770△sasX showed a reduction in cell aggregation,while the complement had no difference with wild type in aggregation. HS770 △sasX showed a significant reduction of adherence to human nasal epithelial cells compared to wild type ( P＜0.01 ),and the complement showed a very clear increasement of adherence to human nasal epithelial cells compared to wild type(P＜0.01 ).Preincubation of nasal epithelial cells with the purified recombinant SasX protein inhibited S.aureus binding significantly.Conclusion SasX had an influence in aggregation of S.aureus and its adherence to human nasal epithelial cells.By acquiring sasX,S.aureus colonized more easily to the susceptible sites of the host,and thus caused infection.

Objective To observe the diurnal variation of intraocular pressure(IOP)measured with Goldmann applanation tonomoter(GAT)in patients with primary open angle glaucoma(POAG),normal tension glaucoma(NTG)and healthy controls,and to compare the differences of diurnal variation curves between two eyes of each subject and define the distribution of the peak of IOP. Design Prospective case series.Participants POAG patients,NTG patients and healthy controls.Methods The diurnal variation of IOP in the two eyes of each of 30 POAG patients,30 NTG patients and 30 normal controls were investigated and the distributions of the peak IOP were observed.Main Outcome Measure Intraocular pressure.Results Measured with GAT,6.7% right eyes and 10.0% left eyes of normal people,20.0% right eyes and 23.3% left eyes of NTG patients,and 23.3% right eyes and 20.0% left eyes of POAG patients showed peak IOP at the time points of out-office period.Conclusions POAG patients,NTG patients and normal controls have asymmetric IOP diurnal variation.The two eyes of one individual cannot always be treated as the same.The IOP peak can occur at the time point during the out-office time.Measure IOP in office-time only may miss the dynamic information of IOP.