Objective To clone lexA gene,express and purify the repressor LexA from Pseudomonas aeruginosa(PA),prepare the polyclonal antibody against PC-1 protein in rabbits,detect immunological activity of LexA protein.Methods The genomic DNA was extracted from the PAO1,the gene fragment encoding the mature LexA was amplied by PCR.It was linked the vector pET32a(+)and expressed in the E.coli BL21(DE3).The expressed protein was purified by two steps of Ni2+ chelate affinity chromatography and gel filtration chromatography respectively.The purified LexA protein immune the rabbits by injection and prepare the polyclonal antibody against PC-1 protein.The immunological activity of expressed and purified LexA protein was detected by ELISA,and Western blot.Results The expressed fused protein was found in insoluble form,accounted for 45% of the total bacteria protein.The final purity was 98.97%,which was determined by the HPLC.The expressed and purified LexA protein had satisfactory immunological activity.

Objective To determine whether the cross-sectional area (CSA)of the calf measured with MRI could stage lower ex-tremity lymphedema (LEL)secondary to gynecological cancer treatments.Methods 148 patients were enrolled in this research.116 females with unilateral LEL and 32 without LEL after gynecological cancer treatments underwent calf MRI and water displacement. Total soft tissue CSA (T),muscle CSA (M)and subcutaneous tissue CSA (S)of affected calf,and difference of T (DT),M (DM) and S (DS)between calves were obtained on MRI at mid-calf level.Volume of affected calf and difference of volume (DV)between calves were obtained by water displacement.Statistical analysis was performed to determine feasibility of MRI measurements for ac-cessing LEL.Results There were close correlations between volume and T or S of affected calf,and between DV and DT or DS of calves.The correlations of stages of LEL with T and S of affected calf as well as DT and DS were stronger than the volume of affect-ed calf and DV (P< 0.01).Multivariate analysis showed more significant differences in T and S than in volume of affected calf,and in DS than in DV between LEL stages (P< 0.05).No difference was found in volume of affected calf and in DV between stage 0 andⅠ. For staging LEL,DS showed the most profound discrimination ability among all measurable parameters.Conclusion DS of calves could be the most reliable parameter recommended for staging and early diagnosis of LEL.

Aim To study the effects of a novel psychoactive drug,modafinil,on the GABAA receptor-induced current in rat hippocampus pyramidal neurons.Methods Whole-cell patch-clamp recordings were applied in cultured hippocampus pyramidal neurons.Results The GABA-activated currents were blocked by bicuculline,a competitive antagonist of GABAA receptor.Modafinil could partly inhibit IGABA.Pre-treating hippocampus neurons with glibenclamide for 30 min,modafinil-induced inhibition on IGABA was removed.Conclusion Modafinil is different from other psychostimulants.The protection of modafinil may be via modulating ATP sensitive potassium channel activation.

Objective To investigate the protection of Liuwei Dihuang Jiawei Capsula(LDJ Capsula,Rehmanniae Capsula of Six Ingredients) on diabetic nephropathy(DN) rats′ kidney and effect on renal protein kinase C(PKC) activity and connective tissue growth factor(CTGF) of DN rats.Methods The DN rat models were induced by ip injection of streptozotocin(STZ).The rats were randomly divided into five groups: control group; DN model group;Lotensin group;LDJ Capsula group;and Lotensis and LDJ Capsula combination group.Drug intervention term was 12 weeks.Renal ultrastructure was observed by transmission electron microscope and Masson staining.Relative kidney weight,blood glucose level,serum and urine creatinine content,creatinine clearance,excretion rate of the 24 hour urine protein,renal PKC activity,and CTGF expression in renal cortex were measured by immunohistochemistry.Results Deposition of collagen in renal of DN rats was conspicuous.Relative kidney weight,blood glucose level,serum and urine creatinine content,creatinine clearance,excretion rate of 24 h urine protein,renal PKC activity and CTGF expression of DN rats increased obviously.All Lotensin,LDJ Capsula,and the combination of these two drugs could decrease renal PKC activity and CTGF expression and ameliorate proteinuria and renal function of DN rats.At the same time they all could abate the deposition of collagen in renal of DN rats.Combination of these two drugs could decrease renal PKC activity and CTGF expression more ob-viously and at the same time had more notable protective effect on kidney of DN rats.Conclusion All Lotensin,LDJ Capsula,and the combination of these two drugs could protect kidney of DN rats.The combination of these two drugs has more obviously protective effect than using Lotensin only.

Objective:To optimize the renaturation procedure of denatured LexA,prepare the repressor LexA from Pseudomonas aeruginosa(PA),which have the satisfactory biologic activity.Methods:The LexA was renatured by the GSH/GSSG dilution method,and the renatured protein were purified by Ni2+ chelate affinity chromatography and gel filtration chromatography,following desalination by Sephadex G25 gel column.The renaturation result were detected by the native polyacrylamide gel electrophoresis and RPHPLC.The immunological activity of all LexA proteins,including the denatured,renatured protein and the renatured protein that was treated with the DTT,were determined by Western blot.Results:The renatured LexA appears both monomer and multimer,which is confirmed by the native polyacrylamide gel electrophoresis analysis and RPHPLC.Gel retardation experiments shows that the renatured LexA have satisfactory biologic activity.

OBJECTIVETo study the efficacy of Xiaoyao Powder (XYP) combined with interferon alpha (IFN-alpha) in treating HBeAg positive chronic hepatitis B (CHB) patients and the effect on their quality of life (QOL).

METHODSTotally 193 patients with HBeAg-positive CHB confirmed by liver biopsy were randomly assigned to 2 groups, Group A (94 cases) and Group B (99 cases). IFN-alpha1b was subcutaneously injected to patients in Group A at the dose of 50 microg, thrice per week. Those in Group B additionally took XYP. The therapeutic course for all was 24 weeks. Clinical efficacy was observed by assessing ALT restoration rate, HBeAg negative rate, HBeAg conversion rate, HBV DNA negative rate, complete response rate, partial response rate, and symptoms integral. The evaluation of QOL was performed by using chronic liver disease questionnaire (CLDQ) score. Adverse reaction occurrence rate was observed in the two groups.

RESULTSBetter effects were obtained in Group A on ALT restoration rate, HBeAg negative rate, HBV DNA negative rate, complete response rate, partial response rate, TCM symptoms integral, the total effective rate of TCM sysmptoms, CLDQ score, and adverse reaction rates, showing statistical difference when compared with Group B (P < 0.05, P < 0.01).

CONCLUSIONXYP could elevate the efficacy of TCM symptoms of HBeAg-positive CHB patients and anti-viral effect, improve their QOL, and reduce adverse reaction of IFN-alpha.

Adult ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Hepatitis B e Antigens ; Hepatitis B, Chronic ; drug therapy ; Humans ; Interferon-alpha ; therapeutic use ; Male ; Quality of Life ; Treatment Outcome

Objective To evaluate endovascular therapy and open surgery for subclavian artery occlusion disease. Methods In this study, 69 patients received endovascular therapy (44 patients)or open surgery(25 patients)from January 2002 to July 2007.Balloon dilatation was carried out in 3 cases and 43 stents was placed in 41 cases. Results All procedures were successful. In endovascular therapy group, the ratio of healthy/dieased side of mean blood pressure was improved from 0.66±0.14 to 0.96±0.13(t=9.532,P＜0.001＝;in surgery group, the ratio improved from 0.63±0.16 to 0.95±0.18(t=8.236,P＜0.001＝.Sixty-one discharged patients were followed up for 2～49 months(mean 16.7 months),in endovascular group, restenosis occurred in 1 patient 1 year after the therapy, in surgery group, all prothesis remained patent and there was no complication related to prothesis. Conclusions Both endovascular therapy and surgery were the effective methods for subclavian artery occlusion, and endovascular therapy is preferred for less invasiveness.

Objective To investigate whether a novel long-acting tumor necrotic factor (TNF) antagonist (soluble TNF receptor:IgG Fc [sTNFR:IgG-Fc]) can protect hepatocyte damage against liver failure caused by drugs in immunity-induced cirrhotic rats.Methods Wistar rats were repeatedly sensitized by human serum albumin (HSA) emulsified in complete freud adjuvant.The blood was collected at day 10 after the final sensitization.If anti-albumin antibody was positive,the rats were intravenously injected with HSA twice a week.After six weeks,liver cirrhosis was induced by immunity.All the model rats were divided into three groups with 15 each.Liver failure was induced with D-galactosamine/ lipopolysaccharide (LPS) intraperitoneal injection in the rats with liver cirrhosis in model group.The rats in pretreatment group were intraperitoneally injected with long-acting soluble TNF receptor p55 18 h before D-galactosamine/LPS injection.The control group were injected with 0.9％ sodium chloride.General condition,survival rate,liver function and pathological changes were all examined.Serum levels of interleukin (IL)-6,IL-22 and intrahepatic level of IL-6 were detected by enzyme linked immunosorbent assay (ELISA).The activity of Caspase 3 in hepatocyte lysis solution was measured by spectrophotography.Real-time polymerase chain reaction (PCR) was used to detect mRNA expressions of proliferating cell nuclear antigen (PCNA),bcl-2,bax and IL-22 receptor.Data were analyzed by variance analysis among groups.Results Rats in model group were dispirited with poor response after 12 hours and only 3 survived,compared with soluble TNF receptor p55 pre-treated group rats,in which all survived (P=0.029 8) with flexible response.Serum alanine aminotransferase levels in these two groups were (6 533± 360) and (105 ± 7) U/L,respectively.Hepatic regenerative nodule developed massive or submassive necrosis with septal fibrosis in model group,whereas soluble TNF receptor p55 alleviated the inflammatory and necrosis reaction of hepatic tissue.Serum IL-6 levels in model group and pretreatment group were (842.0±12.9) and (91.9±1.6) pg/mL,respectively (F=380.30,P＜0.01).Intrahepatic levels of IL-6 in these two groups were (26.2±1.2) and (11.1±0.8) pg/mL,respectively (F=176.90,P＜0.01),and serum IL-22 levels were (167.0±27.8) and (988.0±109.6) pg/mL,respectively (F=37.91,P＜0.01).Hepatic Caspase-3 activity was reduced by almost 60％ by soluble TNF receptor p55 pretreatment (F=303.70,P＜0.01) and bax expression reduced by 22％ (F=108.80,P＜0.01),while bcl-2 and PCNA expressions were up-regulated by 3.6-folds and 23.0-folds,respectively (F=115.60,P＜0.01; F=594.20,P＜0.01).Conclusions Long acting soluble TNF receptor p55 could improve survival rate,liver function and reduce inflammatory reaction of rats with liver failure induced by drugs on the basis of liver cirrhosis caused by immunity,which indicates that this drug may process a potential therapeutic value.

Objective To evaluate the MR lymphangiography (MRL)in diagnosis of limb lymphedema.Methods A total of 582 patients with lymphedemtous limbs were enrolled in the study,MRL was performed at 3.0T MR.The morphology and enhancement of the lymph nodes,the number of lymphatic vessels and the lymph flow were evaluated.Results No matter in primary or secondary lymphedema,there were patients showed only lymph nodes affected,or only lymph vessels affected,and some patients showed both affected.Lymphatic aplasia,hypoplasia or hyperplasia were showed in primary lymphedema.Obstruction lymphatic vessels,and lym-phangiectasia were showed in secondary lymphedema.The velocity of lymph flow was (1.0±0.62)cm/min in affected limb of pa-tients with primary lymphedema,which was significantly slower than that of affected limb of patients with secondary lymphedema (2.22±1.64)cm/min(P<0.01)in dynamic contrast-enhanced MRL.In both type of lymphedema,the contrast enhanced lymph nodes showed less nodes with delayed enhancement and lower signal intensity,compared to that of lymph nodes in the contralateral normal side.Conclusion Dynamic contrast-enhanced MRL is helpful for assessing the anatomical and functional status of lymphatic system in lymphedematous limb.This new imaging techniques provides a powerful tool for the diagnosis of lymphedema.

Objective To explore the optimal cut-off points of body mass index(BMI),percentage of body fat(PBF) and body fat mass index(BFMI) for identification of cardiovascular risk factors clustering among elderly males. MethodsThe data of physical examinations from 1 052 Shanghai elderly males in 2007 were collected.The relationship between cardiovascular risk factors clustering and different strata of BMI,PBF and BFMI was analyzed.Receiver Operator Characteristic(ROC) curve analysis was employed to determine the optimal cut-points for identification of cardiovascular risk factors clustering,and area under curve(AUC) was worked out.The population attributable risk proportion(PARP) of risk factors clustering was calculated. Results Odds ratios of risk factors clustering tended to increase with BMI,PBF and BFMI.BMI≥24 kg/m2,PBF≥21% and BFMI≥5 kg/m2 were the cut-off points that had approximate sensitivity and specificity,and/or had the shortest distance in ROC curve.AUC of all the three indexes was larger than 0.5.Analysis of PARP indicated that BMI under 24 kg/m2,PBF under 21% and BFMI under 5 kg/m2 could prevent 27.1%,37.44% and 36.63% risk factors clustering,respectively. Conclusion BMI≥24 kg/m2,PBF≥21% and BFMI≥5 kg/m2 can well reflect the cardiovascular risk factors clustering among elderly males.