Objective: To investigate the effect of amiloride on in vitro invasive activity and uPA (urokinase-type plasminogen activator)system of human highly metastatic lung carcinoma cell line PGCL3. Methods: At 6 hours after treatment with amiloride at the concentrations of 25μmol/L，50μmol/L and 100μmol/L for PGCL3 cells，Transwell Chamber assay was performed to detect the effect of amiloride on the invasive and migratory capacity of PGCL3 cells.Effect of amiloride on the activity of uPA and PAI-1(plasminogen activator inhibitor-1)secreted by PGCL3 cells were measured by chromogenic substrate assay after PGCL3 cells were incubated with amiloride for 24 hours.RT-PCR was used to analyze the effect of amilorede on mRNA levels of uPA，uPAR(urokinase-type plasminogen activator receptor)and PAI-1.The expression levels of uPA，ERK2(extracellular regulated protein kinases 2)and ras protein were assessed by Western blot. Results: The number of cells through membrane was significantly decreased in invasion and migration test in vitro.The inhibitory rates of invasion and migration after treatment with amiloride of 100μmol/L were 37.7%±4.1%and 64.9%±4.9%.respectively,with a significant difference from those in the control group(P<0.01).At 24 hours after amiloride treatment，the chromogenic substrate assay showed direct inhibition of the activity of uPA and PAI-1 secreted by PGCL3 cells.No effect on the expression of uPAR in mRNA level was observed,but the expression of PAI-1 in mRNA level was significantly inhibited.Amiloride of 100μmol/L dramatically inhibited the expression of uPA mRNA.The expression level of uPA protein was decreased with the increase of the concentration of amiloride,but no effect was observed on the expression of ERK2 and ras in protein level.Conclusion: Amiloride can inhibit the invasion and migration of PGCL3 cells,through inhibiting the expression and activity of uPA and PAI-1.Amiloride is a potential agent to inhibit cancer invasion and metastasis.

Objective:To investigate the effect of dimethyl amiloride (DMA) on invasive activity of PGCL3 cells from a human highly-metastatic lung carcinoma cell line in vitro and elucidate its possible mechanism. Methods:The invasion and migration capacities of PGCL3 cells were measured by using Transwell chamber assay after pretreatment with DMA. The effects of DMA on the activity of urokinase-type plasminogen activator (uPA) and plasminogen activator inhibitor-1 (PAI-1) secreted by PGCL3 cells were measured by chromogenic substrate assay. The effects of DMA on uPA, urokinase-type plasminogen activator receptor (uPAR), and PAI-1 mRNAs transcription were determined by RT-PCR. The expression levels of extracellular regulated protein kinases 2 (ERK2) and ras protein were assessed by Western blot. Results:DMA inhibited invasion and migration capabilities of PGCL3 cells in vitro, down-regulated the mRNA transcription of uPA, uPAR and PAI-1, as well as up-regulated the expression of ras protein. After 24-hour treatment, DMA reduced the activity of uPA at higher concentration, but DMA had no effects on the activity of secreted PAI-1 protein and expression of ERK2 protein. Conclusion:DMA inhibits the invasion and migration of highly-metastatic lung cancer PGCL3 cells. The mechanism might be associated with down-regulation of the expression of uPA system.

OBJECTIVE To explore the resistance of Pseudomonas aeruginosa isolated in clinic against five antiseptics involving in povidone iodine(Iodophor),glutaraldehyde,chlorhexidine,symclosene(trichloroisocyanurate) and benzalkonium bromide.METHODS The susceptibility test of P.aeruginosa was determined by PhoenixTM-100 system.Minimun inhibitory concentration(MIC) of povidone iodine,glutaraldehyde,chlorhexidine,symclosene and benzalkonium bromide was detected by liquid dilution method.RESULTS The resistant rates of ampicillin/sulbactam,chloramphenicol,tetracycline and trimethoprim-sulfamethoxazole in 190 isolates of P.aeruginosa were all more than 98.0%.However,P.aeruginosa was to imipenem and meropenem were 15.3% and 6.8%.It was found that P.aeruginosa possessed the most resistant to glutaraldehyde and symclosene with its MIC50 being 32 ?g/ml and 64 ?g/ml.But the MIC50 of chlorhexidine and benzalkonium bromide were only 1 ?g/ml and 2.4 ?g/ml.Meanwhile,time-kill assays indicated that chlorhexidine could still produce rapid and powerful bactericidal effects at a concentration of 1MIC after 10 min treatment.CONCLUSIONS There are distinct differences in P.aeruginosa against povidone iodine,glutaraldehyde,chlorhexidine,symclosene and benzalkonium bromide.It is very important that antiseptics should be used rationally.Measurements should be taken to decrease dissemination of resistant bacteria and prevent nosocomial infection.

Objective To investigate the clinical values of NT-proBNP in left ventricular enlargement(LVE) and left ventricular dysfunction(LVD) of the patients with essential hypertension(EH) to provide a diagnostic basis for their diagnosis .Methods Plas-ma NT-proBNP concentrations in 120 patients with EH and in 29 normal controls were measured ,then the echocardiography exami-nation was performed to determine the left ventricular ejection fraction ( LVEF) ,left ventricular end-diastolic diameter(LVEDD) , left atrium(LA) and left ventricular systolic diameter(LVSDD) .The correlation between plasma NT-proBNP with LVE and LVD was analyzed .The diagnostic accuracy of NT-proBNP was analyzed by using receiver operating characteristic (ROC) curve .Results There were no statistically significant differences in the aspects of age ,sex and serum creatinine between the EH group and con-trol group ,but plasma NT-proBNP level of the former was significantly higher than that of the latter .The NT-proBNP level in the patients with LVE was significantly higher than that in the patients with normal left ventricle (P<0 .05) .The NT-proBNP level in the LVD patients was significantly higher than that in the patients with normal left ventricular function (P<0 .05) .The plasma NT-p roBNP level was positively correlated with LA ,LVEDD and LVSDD(r=0 .518 ,0 .58 ,0 .48 ,P<0 .01) while negatively crrelated with LVEF(r= -0 .61 ,P<0 .01) .The ROC curve showed that when the NT-proBNP was set at 380 pg/mL ,the sensitivity and specificity of NT-proBNP for diagnosing LVE were 80 .6% and 72 .4% ;which for diagnosing LVD were 80 .8% and 77 .4% ,re-spectively .Conclusion NT-proBNP is closely correlated with multiple ultrasonic indicators reflecting the left ventricular function and its level can reliably reflect the left ventricular contraction function ,which can serve as the marker for screening LVE and LVD in the patients with EH .

Objective: To study the pharmacological action and clinical curative effect of Linglongqingre microenema. Methods: The actions of relieving fever and calming hypnotism. anti convulsions of Linglongqingre Microenema on rabbit, mouse and rat were observed. Also children's fever illess was treated to judge preliminary curative effect. Results: Linglongqingre Microenema has obvious action of relieving fever, more effective than Chaihu injection, as same as Antipyrine (80mg/kg) on subcutaneous injection. It was used to treat children's feverillness on 38 cases, markedly effective rate 60.53%, total effective rate 86.84%. Among them body temperature of one third case fell over 1.5?C in two hours. Conclusion: Linglongqingre microenema is a new quick effective pharmaceutical preparation which is suitable for emergency treatment of traditional Chinese medicine

Objective To investigate the potential application of human transforming growth factor-beta-1 (hTGF-β1) gene mediated by type 2 recombinant adeno-associated virus (rAAV2) vector inducing chondrogenic differentiation of canine mesenchymal stem cells (MSCs) in vitro. Methods Canine MSCs from bone marrow were isolated and cultured in vitro by density gradient centrifngation and adherence screening methods. The morphology of MSCs was observed by inverted phase contrast microscope and Giemsa stain. Flow eytometry was used to detect surface antigens of MSCs, The third generation of MSCs were transfected by rAAV2-hTGF-β1 with or without MOI of 1 ×105 v.g./cell or 5×105 v.g./cell. The expression of hTGF-β1 was detected by Western blot after 10 days, and TGF-β1 synthesis was determined by ELISA at 3, 6 and 9 day, respectively. After 2 weeks of culturing, mRNA expressions of type Ⅱ collagen and aggrecan were determined by RT-PCR and the collagen Ⅱ protein was detected by immunocytochemistry. Results The MSCs appeared to be morphologically spindle-shaped and showed active capability of proliferation both in primary and passage generations. Flow cytometry analysis indicated that MSCs were universally positive for CD29, CD44 and CD105, but negative for CD34 and CD45. TGF-β1 expression can be observed by Western blot after 10 days in two transfection groups, MOI of 5 × 105 group and MOI of 1× 105 group. With the extension of time, the contents of hTGF-β1 increased in the two groups detected by ELISA, while there was a significant difference between them two (P < 0.01). After 2 weeks of transfection of MSCs by rAAV2-hTGF-β1, the expression of collagen Ⅱ and Aggreacan mRNAs were positive. It also showed positive of collagen Ⅱ detected by immunocytochemistry. Conclusion Canine MSCs show chondrogenesis differentiation after induction by Type 2 rAAV mediated transfer of TGF-β1 gene. The process is a potential application for cartilage tissue engineering.

OBJECTIVE:To optimize the extraction technology of asiaticosides from Centella asiatica. METHODS:Using to-tal amounts of asiaticoside and hydroxy asiaticoside as investigation indexes,single factor and orthogonal test were used to investi-gate the enzyme amount,enzymolysis time,enzymolysis temperature,ethanol volume fraction,liquid material ratio and ultrasonic extraction time and optimize the extraction technology of asiaticosides from C. asiatica by ultrasonic-enzyme method,and verifica-tion test was conducted. RESULTS:Optimal extraction technology was as follow as cellulose dosage of 12 mg/g,10-fold liquid ma-terial ratio added into 60% ethanol,enzyme hydrolysis for 60 min at 60 ℃,ultrasonic assisted extraction for 50 min. Average ex-traction rate of total asiaticosides was 1.92%(RSD=1.83%,n=3)in verification test. CONCLUSIONS:Ultrasonic-enzyme meth-od is stable and feasible for the extraction technology of asiaticosides from C. asiatica.

To study the effects of rCBF and brain function in the patients with cerebral infarction byearly rehabilitation training. 89 cases were randomized into rehabilitation and control groups and were ex-amined rCBF by 133Xe inhalation method and BEAM. Total effect rate was 93.9%in rehabilitationgroup,to the control 77.5%(X2=3. 95,P<0.05). The rCBF rised up in two groups,but it was higher inthe foriner,to the contro1,t=4. 99,P<0. 01. BEAM improve rate was 95.9%,to the control,77.5%(X2=5. 30,P<0. 05). So we confirmed that early rehabilitation training may promote rCBF and improve brainfunction of patients with cerebral infarction.

Objective To investigate the prevalence rate of metabolic syndrome (MS) and its related diseases in Bengbu community residents. Methods 3246 residents aged from 25 to 74 years were chosen through cluster random sampling method, including 1459 males and 1787 females. MS and its related diseases were examined. Results (1) The prevalence rate and standardized rate of MS was 20. 5% and 16.0%, respectively, which reached higher level all over China. The prevalence rate of male and female was 22. 9% and 18. 6%, respectively. The highest prevalence rate occurred in presenium patients. (2)The prevalences of various metabolic diseases were increased with aging except for overweigh/obesity and MS (P<0.01). The prevalence of various metabolic diseases was 1 times higher in elderly patients than that in young patients, and the prevalence rate of MS in male and female increased to 2.8 times and 2.7 times, respectively. The prevalence rate of type 2 diabetes mellitus increased biggest, male to 7.9 times and female to 12.8 times compared with the original level. (3) The multiple components clustering rate of MS was high, only 33.8% of individuals had no metabolic disorder. Conclusions The prevalence rates of MS and its related diseases are high in Bengbu community residents. It is necessary to take comprehensive measures to prevent and control MS in community.

Objective To ensure the medical security of the astronauts,new targeted strategies were adopted after summarizing the experience in Chinese astronauts rescue and medical aid at the main landing site,the specialty and characteristics of landing were analysied.Methods Search the publications about astronaut medical aid domestic and abroad,summarize the rescue and medical aid experiences from Shenzhou 5 to Shenzhou 10.In consideration of prolonged on-orbit time,the cold weather conditions at the landing zone of Shenzhou 11,new targeted strategies were presented.Results On the basis of the original helicopter emergency platform and first aid equipment,the emergency aid procedures were optimized,personal warm clothing,a heat preservation box,insulation blanket,self-heating pads and intraosseous rapid infusion system were used to ensure the medical security of astronauts in cold weather at the main landing site.Conclusions With the procedures optimized and the targeted strategies performed,the astronauts' s rescue and medical aid project was fully meet the cold and complex conditions at main landing site.