Objective To determine the free thiols in the chimeric protein PfCP-2. 9 of Plasmodium falciparum expressed by Pichia pastoris. Methods Two experiments of reverse phase HPLC and Ellman' s reaction were applied to the PfCP-2.9 for the determination of its free thiols. For RP-HPLC analysis, three kinds of samples were tested: PfCP-2. 9, dithiothreitol-reduced PfCP-2.9 and indoacetic acid-alkylated PfCP-2.9. Results Both experiments showed that there were no any free thiols present in the PfCP-2. 9. Conclution The disulfide bonds between cysteine residues of PfCP-2. 9 were formed completely.

Objective:To express MSP1-31 gene of Plasmodium falciparum in Salmonella typhi CVD908 vaccine strain using a tetracycline-controlled P LtetO promoter. Methods:The MSP1-31 gene was cloned into the plasmid of pZE11 and transformed into the CVD908/tetR strain by electroporation. Expression of MSP1-31 in CVD908/tetR strain was detected using the method of Western blot. Results: The recombinant plasmid of pZE11/MSP1-31 was constructed, there was effective expression of MSP1-31 protein in CVD908/tetR strain in presence of tetracycline, and no expression of gene in absence of tetracycline. Conclusion: The recombinant Salmonella typhi strain in which the expression of Plasmodium falciparum MSP1-31 fragment induced by tetracycline is established successfully.

OBJECTIVE:To develop an LC-MS method for the determination of the concentration of duloxetine in human plasma.METHODS:Plasma sample(0.2 mL)was taken,with the protein precipitated twice using acetonitrile.The mobile phase consisted of 20 mmol?L-1 ammonium acetate(pH3.5)-methanol-acetonitrile(38∶20∶42).The sample was separated on a Thermo C18 column with plasma concentrations(time-concentration profile)of duloxetine in 10 healthy volunteers determined by LC-MS after taking a single oral dose of duloxetine enteric-capsule(22.4,44.8 or 67.2 mg).RESULTS:The calibration curve of duloxetine was linear over the concentration range of 0.78～100 ng?mL-1(r=0.999 5)with the recovery at 100.8%,the RSD of intra-day and inter-day at no more than 7.8% and 9.3%,respectively.CONCLUSION:The method developed in our study was proved to be accurate and sensitive,and suitable for the pharmacokinetic study of duloxetine.

arotid artery atherosclerosis is closely associated with ischemic stroke.Carotid artery intimamedia thickness is an important indicator of carotid artery atherosclerosis.Magnesium is involved in the in vivo metabolism of a wide variety of processes.Studies have shown that magnesium can decrease carotid intimamedia thickness.This article reviews the relationship between magnesium levels and carotid artery atherosclerosis in different populations.

Surgical treatment is the treatment of choice for gastrointestinal stromal tumor (GIST).A consensus on the surgical principle of GIST has been made based on the 10-year experience,which effectively guided the standardized treatment of GIST.Laparoscopic and gastrointestinal endoscopic techniques were applied in the treatment of GIST,and promote the development of the diagnosis and treatment of GIST.Laparoscopic treatment has become one of the main treatment methods for GIST.Furthermore,multi-disciplinary treatment for GIST is gradually adopted in clinical practice.In this article,the prospect of surgical treatment of GIST is introduced based on a 10-year experience.

The paper introduced the practice of family doctor′s contracted service in Zhenjiang,Jiangsu province over the past six years.By exploring the tiered family doctor′s contracted services,namely 3+X teamwork responsible for household health,health insurance contracting with chronic disease patients,and personalized contracts,the city highlights the cornerstone role of primary care in the Healthy Zhenjiang 2015 Campaign.The campaign aims at providing fair,accessible,systematic and uninterrupted health services.Outcomes of the campaign have proved that the 3+X teamwork model helps building acquaintances relationship with residents by means of contracting,hence motivating both the sector and individuals in the health promotion and building a healthy Zhenjiang.

Objective:To observe the effect of Tangshenning on renal function and LOX-1 mRNA expression in the kidney of early diabetic nephropathy rats model. Methods:Early diabetic nephropathy rats model was made with high fat diet, STZ and unilateral ne-phregtomy. After the 12-week drug intervention, the rats were sacrificed, the kidneys were removed and the mRNA expressions of LOX-1 in the kidney were observed by the method of RT-PCR. Results:Early diabetic nephropathy rats model was successfully made by the triple-modeling method. The mRNA expression of LOX-1 in the kidney was significantly increased, and Tangshenning could low-er the expression of LOX-1 in the kidney in a dose-dependent manner. Conclusion:The abnormal expression of LOX-1 could be one of the mechanisms for diabetic nephropathy. Tangshening has good kidney protective effect through decreasing LOX-1 abnormal expression in the kidneys, which deserves further research.

Objective To map out the binding site of P195,which is the major protein on the surface of P.falciparum merozoites,to human erythrocytes,and offer a basis for designing malaria vaccine to blockade invasion of merozoites into human erythrocytes.Methods Eight proteins derived from P195 were expressed in E.coli,and purified by Ni-chelare affinity chromatography.There after,the eight fragments and rabbit serums immunized by which were added into culture medium of P.fatciparum in vitro respectively.Twenty-four hours later,the invasion of merozoite to erythrocyte was observed.Results The antibodies which were induced by three fragments of P195,M6(Amino Acid,AA384～595),M7(AA 595～897)and M11(AA 1397～1563)could inhibit the invasion of P.falciparum merozoite into human erythrocytes.Especially,one fragment of P195,M6,had the ability to inhibit the invasion of P.falciparum merozoite into human erythrocytes.Conclusion M6,a fragment of P195 on the merozoite of P.falciparum may contain a domain thought to be involved in the recognition of human erythrocyte.The domain can be used as a candidate antigen for a malaria vaccine.

Objective To investigate the effect of TanshinoneⅡA (TSN) on the cell hypertrophy induced by angiotensinⅡ(AngⅡ) in the primary culture of neonatal rat cardiomyocytes. Method The effect of TSN on cardiomyocyte was evaluated by the 3-[4, 5-dimethylthiazol-2-yl] -3, 5-diphenylformazan (MTF) assay. As the index of eardiomyocyte hypertrophy, protein synthesis rate was measured by H-Leucine incorporation and the cell size was determined by phase contrast microscope. The proto-oncogene c-los mRNA and c-jun mRNA expression was assessed using reverse transcription polymerase chain reaction (RT-PCR). Results Exposure of the myocytes to TSN (5～80 mmol/L) for 24hours produced no cytotoxicity. Protein synthesis rate and proto-oneogene c-fos and c-Jun mRNA expression of eardinmyoeytes increased significantly after AngⅡtreatment, and TSN inhibited these effect of AngⅡ.Conclusions TSN can prevent the hypertrophy of eardiomyocytes induced by AngⅡ, which be attributable relate to the decreased expression of proto-oncogene c-los and c-jun mRNA by TSN.

Objective To investigate the effects of Astragalus polysaccarides (APS) on 2,4,6-trinitrobenzenesulfonic acid (TNBS)-induced colitis in rats. Methods Forty male Sprague-Dawley rats were randomly divided into five groups (n = 8 ): control group, TNBS group, low-dose APS group, high-dose APS group, and prednisone group. Experimental colitis was induced in rats by enema administration of TNBS. Rats in APS and sions and histological damages were determined, and the activity of myeloperoxidase (MPO) was measured in the excised colonic tissues. Cytokine levels including interleukin (IL)-4 and IL-10 were determined by enzyme-linked immunosorbent assay. Results Both macroscopic lesions and histological colonic damages induced by TNBS were reduced by low-dose APS treatment. These were accompanied by significantly attenuated colonic MPO activity (P = 0. 03) and the increase of IL-4 and IL-10 levels. The macroscopic lesions and MPO activities of high-dose APS group were higher than TNBS group, histological damage and level of IL-4 were lower, and level of IL-10 was higher, but all without statistical significance. Levels of IL-4 and IL-10 were lower than those of TNBS group, but there was no significant difference between prednisone group and TNBS group. Levels of IL-4 and IL-10 were significantly lower in prednisone group than in control group ( P = 0. 049, P = 0. 001 ). Conclusions Different doses of APS have different effects on TNBS-induced colitis. Lower dose of APS has the therapeutic potential inexperimental colitis, while higher dose of APS may aggravate the disease.