Objective To optimize the conditions for enhancing the refolding of a novel recombinant human(rh)endostatin and test the biological activities of refolded endostatin.Methods The partial purified inclusion bodies of rh-endostatin were dissolved with 6 mol/L guanidine-HCl followed by combination of dilution and dialysis of the dissolved endostatin.The refolded endostatin was then purified by cation-exchange chromatography.The biological activities of purified rh-endostatin were assessed by endostatin-specific monoclonal antibody and chick embryo chorioallantoic membrane assay.Results A 46% refolding yield was achieved after optimizing the refolding conditions.The purified endostatin reacted with specific anti-endostatin monoclonal antibody and showed significant inhibition of angiogenesis in chick embryo ehorioallantoic membrane assay.Conclusion The method of highest refolding yield of human endostatin was developed.This optimized method significantly promotes the application of this novel human endostatin to preclinical and clinical studies.

This review focus on the perspective of genetic engineering technology in biopharmaceutical field.Four general aspects of genetic engineering are further discussed in this review as following: the separation of macromolecules,PCR,gene chip,and the expression of foreign genes.The large-scale or industrialization of genetic engineering technology in biopharmaceutics is also reviewed.

Objective To analyze the relationship between hepatitis B virus gene mutation at site 1896 in precore region and genotypes as well as liver function parameters. Methods The fluorescent quantitative PCR and sequencing method were applied to measuring the relevant indicators in 50 patients with chronic hepatitis B. Results There was significant difference in ALT level between hepatitis B patients with site 1896 mutation and ones with wild-type; and HBV mutation at site 1896 in precore region was unrelated to the genotypes. Conclusion HBV mutation at site 1896 in precore region may be associated with continous viral invasion invasion into hepatocytes.

BACKGROUND:Al oying is a convenient and effective method to alter the microstructure and control the corrosion behavior of magnesium al oy.
OBJECTIVE:To explore the in vitro and in vivo degradation of Mg-Nd-Zn-Zr al oy as a degradable medical biomaterial.
METHODS:(1) In vitro static immersion test:The immersion tests were carried out at (37.0±0.5) thermostatic bath. Six Mg-Nd-Zn-Zr al oy samples and six pure magnesium samples were immersed in the 250 mL simulated body fluid and vibrated without agitation during immersion. After 3, 7 and 30 days static immersion, the samples were taken out from the simulated body fluid. Then the in vitro corrosion properties were evaluated using scanning electron microscope and energy dispersive spectrometer analysis. (2) In vivo animal experiment:After bone tunnel was established in the left femur of adult New Zealand rabbits, the Mg-Nd-Zn-Zr al oy rods were embedded in the Mg-Nd-Zn-Zr al oy group, titanium al oy rods were embedded in the titanium al oy group, and only bone tunnel was established in the sham-operated group. At 1, 2, 4, 8 weeks after implantation, an X-ray of the implanted region was taken to determine the location and the degradation behavior℃in a of Mg-Nd-Zn-Zr al oy. At 4, 8 weeks after implantation, the corrosion product and its element composition were observed using scanning electron microscope with an energy dispersive spectroscopy system.

Objective To investigate the influence of psychological intervention on the urodynamic test. Method The 959 patients in the urodynamic test were divided into two groups:the control group and the the test group,then we analyzed and compared the results. Results In the incidence of the abnormal abdominal pressure, the unnatural detrusor contraction and the Unnatural urethral sphincter contraction, there were statistically significant differences between two groups ( <0.05) . Conclusion It effective and necessary for urodynamic test to do psychological intervention.

Objective To explore clinical application of coronary artery imaging with 64-slice spiral CT.Methods 46 patients with coronary heart disease were undergone both coronary artery imaging with 64-slice spiral CT and coronary angiography,and the effects on coronary display respectively were compared Results Coronary display in coronary artery imaging with 64-slice spiral CT was better than that of the coronary angiography, which was a inspection pattern of noninvasive coronary display. Conclusion Coronary artery imaging with 64-slice spiral CT can accurately evaluate coronary artery stenosis, which was worth to application.

Objective To explore the treatment effect and comfort of colostomy surgery with fecal water der-matitis around stoma after application of multilayer soft silicone dressings.Methods Approved by the Hospital Ethics Committee and the patients'informed consent,120 patients were divided into the hydrocolloid dressings group ( the control group) and multi-layer soft silicone dressing group (the treatment group),60 cases in each group.Before treatment,after treatment 24h and 36h,DET score,GCO score and the treatment effect were evaluated.Results After treated for 24h,the DET,GCO scores and treatment effects ( in the control group,cured in 3 cases,effective 45 cases, in the treatment group 5 patients were cured,effective in 46 cases) had no significant differences between the two groups (t=-2.624,P>0.05,t=-0.429,P>0.05,χ2 =0.519,P>0.05).After treated for 36h,the DET,GCO scores and treatment effects ( in the control group:cured in 29 cases,effective in 25 cases,in the treatment group:51 patients were cured,effective in 9 cases) had significant differences between the two groups ( t=-5.022,P<0.01,t=12.784,P<0.01,χ2 =6.316,P<0.05).Conclusion Colostomy fecal water dermatitis around stoma after application of multilayer soft silicone dressings can significantly improve the patients'comfort.

Objective: To assess the regional systolic and diastolic function of left ventricle by pulsed wave Doppler tissue imaging ( PW-DTI) in patients with coronary artery disease ( CAD ) . Methods : Seventy-seven cases of coronary angiography were adopted for study and grouped in terms of severity of coronary arterial lesion. Peak systolic, peak early diastolic, and peak late diastolic motion velocities (Sm, Em and Am, respectively) were recorded at 6 different sites on the basal and medial segments corresponding to the anteroseptal, posteroseptal, lateral, anterior, inferior, and posterior walls of the left ventricle with PW-DTI. Results: Am in segments corresponding to normal coronary arteries (group A) was lower than in segments corresponding to slightly stenosed coronaries (group B). There was no significant difference in Sm and Em between group A and group B. Compared with group A, Sm, Em and Am were significantly decreased in segments corresponding to severely stenosed coronary arteries ( group D). Conclusion : PW-DTI could be used to evaluate quantitatively and accurately the regional mvocardial function of left ventricle in patients with CAD.

Objective To identify hepatitis C virus(HCV)-specific cytotoxicity T lymphocytes (CTL)epitopes by the combination of T epitopes prediction software and in vitro assays.Methods HCVspecific CTL epitopes were predicted by T epitope prediction software Rankpep and then candidate HCV-specific CTL epitopes were selected.Candidate HCV-specific CTL epitopes were used to stimulate PBMC of HCV-infected patients and healthy volunteers.and then enzyme-linked immunospot(ELISPOT)and intracellular cytokine staining(ICS)were used to measure the frequencies of IFN-γ-producing cells in total PBMC and the percentages of IFN-γ+CD8+T cells in total CD8+T cells,respectively.Results Five candidate CTL epitopes[NS3 450(TVPQDAVSR),NS3 594(GPTLLYRL),Ns4b 78(sMMAFSAAL),NS5a 416(SEENVSVVF)and NS5a 367(TVSSALAEL)]were used to stimulate PBMC of ten HCV-infected patients and two healthy volunteers.PBMC of seven HCV-infected patients secreted IFN-γ while PBMC of healthy volunteers did not produce IFN-γ.The frequencies of peptide-specific IFN-γ-producing cells ranged from 5 to 36 SFC/105 PBMC and the percentages of peptide-specific IFN-γ+CD8+T cells ranged from 0.02%-0.25%.Conclusion Resuhs of ELISPOT assay and ICS assay confirm that these five peptides NS3 450,NS3 594,NS4b 78,NS5a 416 and NS5a 367 are identified as Hovel HCV-specific CTL epitopes.

Purpose To investigate whether FoxM1 participate in inhibitory effect of cisplatin (CDP) in resistant osteosarcoma cell lines by down-regulation of Rad51.Methods The resistant osteosarcoma cell lines were induced by gradually increasing dose intermittent action,and were named MG-63/R and HOS-MNNG/R respectively.The mRNA and protein level of FoxMl and Rad51 were detected by qRT-PCR and Western blot analysis in resistant cells and parental cells.The mRNA and protein level of FoxM1 and Rad51 were detected by qRT-PCR and Western blot analysis in resistant cells after treatment of 4 μmol/L Thiostrepton.The effect of single or combined treated of 4 tμmol/L Thiostrepton or 2 tμg/mL CDP on the rate of cell proliferation in resistant cells was examed by cell counting.Results Resistant osteosarcoma cell lines MG-63/R and HOSMNNG/R were established and stablely growthed in the concentration of 2 μg/mL CDP,and the resistance index was 30.52 and 37.87 respectively (severe CDP resistance).The mRNA and protein level of FoxM1 and Rad51 were significantly increaced in resistant cells compared with parental cells.The proliferation rate of resistant cells in conbination of 4 μmol/L Thiostrepton and 2 μg/mL CDP treated group was significantly lower than these two drugs single treated group.The level of mRNA and protein of FoxM1 and Rad51 were significantly decreased after 4 μmol/L Thiostrepton treatment in CDP resistant cells.Conclusion The results suggest that FoxM1 and Rad51 may participate in the resistant osteosarcoma cells to CDP.FoxM1 inhibitor Thiostrepton may strengthen the inhibitory effect of CDP in the resistant cells by down-regulation of Rad51.