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.

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.

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 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

To investigate the knowledge, attitude and practice (KAP) on rabies among a mass of the exposed population in Beijing so as to provide scientific evidence for development of measures on the prevention and protection of rabies, the descriptive studies were employed, in which patients were interviewed face to face with a standard questionnaire in hospital or the center for disease control and prevention (CDC), including history of exposure, knowledge on rabies, post-exposure treatment etc. Of 478 respondents, 76% of them were bitten on the upper limbs with the exposure proportions in category Ⅰ, Ⅱ and Ⅲ of 24%, 70% and 6% respectively. The proportions of awareness of knowledge in rabies before biting, its prevention, the correct measure to treat their wounds themselves, rabies vaccine inoculation, and injections with rabies antiserum constituted in 82%, 55%, 59%, 99% and 22% respectively. Result of this investigation indicates that most of the people investigated had a lower awareness of knowledge on rabies, the correct measures to treat rabies after biting as well as the measures to prevent rabies, suggesting that community-based prevention and control measures should be emphasized, especially targeting the population with high risk and the heath education on rabies should performed promptly by several governmental departments.

To analyze features of the rabies epidemic in China between 2007 and 2011,identify factors influencing the epidemic and to provide a scientific basis for further control and prevention of rabies,Descriptive epidemiological methods and statistical analysis was used on data collected from the National Disease Reporting Information System between 2007 to 2011 and the National Active Surveillance System between 2007 and 2010.Our analysis shows that while the number of human rabies cases decreased year by year,the number of districts reporting cases did not show significant change.The situations in Guangdong,Guangxi,Guizhou and Hunan provinces clearly improved over the period but they remain provinces with high-incidence,and consequently influence the epidemic situation of surrounding provinces and possibly the whole country.Summer and autumn were high-incidence seasons.Farmers,students and pre-school children represent the high-risk populations,and rates of cases in farmers increased,those for students decreased,and pre-school children remained unchanged.Provinces with active surveillance programs reported a total of 2346 individual cases,of which 88.53％ were associated with canines.Postexposure prophylaxis (PEP) of rabies cases was not significantly improved,whereas PEP in post-exposure population was good.In rural regions of China,canine density was reduced somewhat,and the immunization rate increased slightly.Finally we show that while the epidemic decreased 2007 to 2011 in China,cases continued to be diffused in certain regions.Lack of standardization of PEP on rabies cases was the main reason of morbidity.The high density and low immunization of dog in rural areas and the defective situation of PEP are still continuous occurrences in China and remain a cause for concern.

Objective To evaluate the using of either 225 or 150 microgrammes of mifepristone combined with misoprostol for termination of second-trimester gestation(16～24 weeks).Methods 180 women requesting voluntary induced abortion during gestation 16～24 weeks were randomised to three groups,group 1:oral mifepris- tone 225rag,group 2:oral mifepristone 150mg,and group 3:injected 100rag rivanot by amniocentestis.The total suc- cess rate,once success rate,the interval of having-medicine to uterine-constraction,the volume of bleeding within 2 hours after labour and cervical laceration rate were observed.Results The once success rate of induced labour in group 1 was higher than that in group 2 and group 3(P

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.