Cloning, expression and purification of novel gene Rv2742 in Mycobacterium tuberculosis H37Rv / 生物工程学报

Rv2742 is a novel gene identified from Mycobacterium tuberculosis H37Rv by the proteogenomics strategy. The aim of this study was to establish a system of soluble expression and purification of the missing protein Rv2742 in M. tuberculosis H37Rv, to provide reference for further research on the biological function of Rv2742. The soluble protein was not successfully induced by prokaryotic expression vectors pGEX-4T-2-Rv2742, pET-32a-Rv2742, pET-28a-Rv2742 and pMAL-c2X-Rv2742. After the codon of novel gene Rv2742 was optimized according to E. coli codon usage frequency, only the recombinant strain containing plasmid pMAL-c2X-Rv2742 could produce soluble products of Rv2742 encoding gene. In addition, the expression effects of the desired fusion protein were also analyzed under different conditions including hosts, culture temperatures and IPTG concentrations. The optimum expression conditions were as follows: Rosetta (DE3) host, 16 °C culture temperature and 0.5 mmol/L IPTG. After being purified by affinity chromatography with amylose resin, the fusion protein sequence was confirmed by LC-MS/MS. These results indicated that the novel gene Rv2742 product could be successfully induced and expressed in a soluble form by the expression system pMAL-c2X with MBP tag. Our findings provide reference for studies on potential interaction and immunogenicity.

Efficacy comparative of different laboratory test reagents for hepatitis C virus antibody / 临床肝胆病杂志

ObjectiveTo investigate the effects of different laboratory test reagents for hepatitis C virus (HCV) antibody through a comparative analysis. MethodsA total of 207 samples which tested positive by four anti-HCV screening reagents commonly used in the laboratories in China (Kehua, Xinchuang, Wantai, and Abbott) were included. HCV RNA nucleic acid amplification (NAT) was performed, and if NAT results were negative, recombinant immunoblot assay (RIBA) was performed for further confirmation. The test results of these four screening reagents were compared, and their S/CO values and true positive rates were analyzed. ResultsOf all the 205 samples testing positive by any one reagent, 191 (93.2%) tested positive by the four reagents, and 14 (6.8%) were tested inconsistently by the four reagents. The positive predictive values of Xinchuang, Kehua, Wantai, and Abbott reagents were 88.2% (180/204), 93.8% (180/192), 91.4% (180/197), and 90.0% (180/200), respectively. The S/CO thresholds with a positive predictive value of ≥95% for Xinchuang, Kehua, Wantai, and Abbott reagents were 9.0, 4.0, 5.0, and 7.0, respectively. ConclusionXinchuang, Kehua, Wantai, and Abbott reagents have significantly different S/CO thresholds with a positive predictive value of ≥95%, which are significantly different from those in other domestic laboratories. Each laboratory should establish an applicable S/CO threshold with a positive predictive value of ≥95%, in order to reduce the sample size for confirmatory test.

Educational reform centered on independent study / 中华医学教育探索杂志

Reform was made on traditional education mode based on the criterion of undergraduate medical education at home and aboard.The reform includes the changes in teaching content,teaching methods and assessment methods in an aim to establish independent learning mode,cultivate students' self-study ability,initiative spirit and innovation ability.

The relationship between LDL oxidation and macrophage myeloperoxidase activity / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To explore low density lipoprotein (LDL) oxidation by macrophage myeloperoxidase (MPO) at molecular level.</p><p><b>METHODS</b>Using a mouse macrophage model, we examined the relationship between LDL oxidation and macrophage MPO by measuring macrophage MPO activity, LDL oxidation products, MPO gene expression and cellular orientation of LDL oxidation.</p><p><b>RESULTS</b>MPO gene expression increased to its maximum gradually when the concentration of LDL was increased, and then maintained at that level. NaN(3) inhibied the elevation of MPO activity and LDL oxidation, which was LDL concentration-dependent. After the composition of macrophage membrane was roughly analyzed, it was determined that the contents of MPO and LDL in 5% sucrose were 7.667 and 21 times higher than those in 10% sucrose, respectively.</p><p><b>CONCLUSION</b>LDL is attached to the "microdomain" of the macrophage membrane in which LDL is oxidized by MPO.</p>

Preparation and Quality Evaluation of Lisinopril Polyvinyl Alcohol Particles / 中国药房

OBJECTIVE:To prepare lisinopril-loaded polyvinyl alcohol(PVA)particles(LIS-PVA-P)and establish its quality control method. METHODS:LIS-PVA-P was prepared by spray-drying process with PVA as carrier. The preparation was detected in terms of morphology,particle size,span,drug-loading capacity,encapsulation and in vitro dissolution. RESULTS:The preparation assumed sphere with porous surface. The average particle size was 17.29 ?m while drug-loading capacity 31.40%,encapsulation 94.20%,Span was 0.88,90% of the drug loads were released within 30 min. CONCLUSION:The preparation process is simple,good in repeatability and qualified in quality.

Study of the relation between low density lipoprotein and macrophage-myeloperoxidase / 中国免疫学杂志

Objective:To study the relation between low density lipoprotein and macrophage myeloperoxidase.Methods:Using the reaction that MPO catalyze the oxidation of o dianisidin hydrochloride,MPO activity was determined.Using reverse transcription polymerase chain reaction(RT PCR),MPO gene expression was determined.The two methods were used to observe the relationship between low density lipoprotein and macrophage myeloperoxidase activity.Results:LDL was able to accelerate the hoist of MPO activity, but its effect is less than LPS OX LDL have no this effect When the time of LDL effect was prolonged, MPO activity was enhancing little by little When the concentration of LDL effect was increased, MPO activity was enhancing to max little by little, then it come to plateau Conclusion:LDL non especially induced the hoist of MPO activity and the swelling of MPO secretion The reason of the hoist of MPO activity was likely to make MPO into action and enhance its secretion, but not MPO gene expression