Material basis and molecular mechanism of Angelicae Sinensis Radix in activating blood:based on computer-aided drug design / 中国中药杂志

Angelicae Sinensis Radix excels in activating blood, but the scientific mechanism has not been systematically analyzed, thus limiting the development of the medicinal. This study employed the computer-aided drug design methods, such as structural similarity-based target reverse prediction, complex network analysis, molecular docking, binding free energy calculation, cluster analysis, and ADMET(absorption, distribution, metabolism, excretion, toxicity) calculation, and enzyme activity assay in vitro, to explore the components and mechanism of Angelicae Sinensis Radix in activating blood. Target reverse prediction and complex network analysis yielded 40 potential anticoagulant targets of the medicinal. Gene Ontology(GO) term enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analysis indicated that the targets mainly acted on the complement and coagulation cascade signaling pathway to exert the anticoagulant function. Among them, the key enzymes thrombin(THR) and coagulation factor Xa(FXa) in coagulation cascade and thrombosis were the drug targets for thromboembolic diseases. At the same time, molecular docking and cluster analysis showed that the medicinal had high selectivity for FXa. According to binding free energy score, 8 potential active components were selected for enzyme activity assay in vitro. The results demonstrated that 8 components inhibited THR and FXa, and the inhibition was stronger on FXa than on THR. The pharmacophore model of 8 active compounds was constructed, which suggested that the components had the common pharmacophore AAHH. The ADMET calculation result indicated that they had good pharmacokinetic properties and were safe. Based on target reverse prediction, complex network analysis, molecular docking and binding free energy calculation, anticoagulant activity in vitro, spatial binding conformation of molecules and targets, pharmacophore model construction, and ADMET calculation, this study preliminarily clarified the material basis and molecular mechanism of Angelicae Sinensis Radix in activating blood from the perspective of big data, and calculated the pharmacology and toxicology parameters of the active components. Our study, for the first time, revealed that the medicinal had obvious selectivity and pertinence for different coagulation proteins, reflecting the unique effect of different Chinese medicinals and the biological basis. Therefore, this study can provide clues for precision application of Angelicae Sinensis Radix and the development of the blood-activating components with modern technology.

Clinical application and comparison of rapid and accurate identification of mycobacterium by gene chip microarray and smear acid-fast staining / 国际检验医学杂志

Objective To compare the clinical effect of application of gene chip microscopy technique for rapid identification of Mycobacterium and classic smear acid-fast staining,and to assess the advantages and disadvantages of the wo methods.Methods From 201 1 to 2014,gene chip microarray and smear acid fast staining were used to identify the mycobacterium tuberculosis in speci-mens suspicious of the infection from all the general hospitals of Shenzhen city.Chi-square test was used to compare the positive rates of the two methods.Results A total of 2 481 specimens were collected from clinic.With smear acid-fast staining technique, the positive specimens of 1 93 cases werefound and the positive rate was 7.8%.Meanwhile,31 7 positive samples were detected by the technology of gene chip microarray,and the positive rate was 12.8%.The positive rate of Gene chip microarray technology was higher than that of the smear acid fast staining,and there was significant difference between them (P < 0.05 ).The 31 7 positive samples identified by Gene chip microarray,included 263 cases of Mycobacterium tuberculosis,27 cases of Mycobacterium absces-sus,18 cases of Mycobacterium intracellulare,3 cases of Mycobacterium gastric uLcer,3 cases of Mycobacterium avium,1 case of Mycobacterium Gordonae,1 case of Mycobacterium marinum and 1 case of Mycobacterium Kansas.Conclusion The gene chip mi-croarray technology is fast,accurate,and its positive rate is higher than that of smear acid-fast staining technique.Classification and identification of Mycobacterium is very helpful for clinical individualized treatment of anti mycobacterium infection.

Research on Clinical Resistance and Aerolysin Genes of Aeromonas Hydrophila / 现代检验医学杂志

Objective To study Aeromonas hydrophila infection and the clinical status of the major causative factor Aerolysin gene.Methods Clinical isolates of Aeromonas hydrophila was collected from 2012 to 2013 year in People’s Hospital of Shenzhen Longhua Branch.Its identification and antibiotic susceptibility testing was analyzed by VITEK 2 compact.Clinical resistance rates and distribution was analyzed by WHONET5.6 software.Polymerase chain reaction (PCR)was used to de-tect the Aerolysin gene form in chromosomes and plasmids extracted genome.Results The clinical total of 48 isolated Aero-monas hydrophila ,distributed in 16 clinical samples of sputum,blood 11,10 secretions,urine 7 and stool 4.Distribution in ward decentralized,the central tendency was not detected.Drug resistance rates to ampicillin,ampicillin/sulbactam and ce-fazolin reached more than 80%,while amikacin,cefepime,levofloxacin,piperacillin/tazobactam and imipenem was low 10%or less.43.7% strains carried the Aerolysin gene.39.5% of Aerolysin gene was found on chromosome genome.Conclusion Aeromonas hydrophila clinical infection existed in dispersed form,most of which carried Aerolysin gene in chromosome ge-nome.Aeromonas hydrophila had serious resistance to penicillins and first generation cephalosporin,but to broad-spectrum drugs maintaining high sensitivity.Precaution of Aeromonas hydrophila ,as an important condition pathogenic bacteria,is some significant for preventing it’s proliferation of drug-resistant strains.

Identification and analysis of mycobacterium isolated from clinical samples in ShenZhen / 国际检验医学杂志

Objective To investigate the status and species of mycobacterial infections in Shenzhen for clinical diagnosis and treatment to provide a reliable scientific basis .Methods 1 096 of samples from patients with suspicious were detection by gene chip .Results Positive rate of microarray detection mycobacterium was 9 .40% (103/1 096) .103 cases of positive were 87 mycobac-teria by gene chip ,and 16 cases of non-tuberculosis(5 cases of M .abscessus ,3 cases of M .intracellulare ,3 cases of M .avium ,2 cases of M .fortuitum ,1 cases of M .kansas ,1 case of M .marinum ,1 case of M .gordonae .Conclusion The mycobacterial infections in Shenzhen ,tuberculosis infection as the main disease types (84 .47% ) .Non-tuberculous mycobacteria′s isolation ratio has reached 15 .53% ,including 7 kinds of species infection and one case of mixed infections .Identification of M ycobacterium by genechip have great significances for individualized treatment .

Cloning and expression of a new glucoamylase gene / 生物工程学报

According to the reported gene sequence of Rhizopus oryzae glucoamylases, the glucoamylase gene containing four introns was cloned from the total DNA of the natural Rhizopus arrhizu. Specific primers were designed to delete introns by overlapping PCR and a new cDNA sequence of Rhizopus arrhizu glucoamylase was obtained. The accession number in gene bank is DQ903853. This gene is successfully expressed in the Picha pastoris, producing a new protein with a high activity of glucoamylase.