1.Identification and renaming of one strain used for tracheitis vaccine production
Jichun SHI ; Huijing DU ; Li LIANG ; Kang LI ; Xiao XU ; Yuhua XING ; Cuiping CHENG ; Qiang YE
Chinese Journal of Microbiology and Immunology 2016;36(6):448-452
Objective To identify and rename one strain stored in National Center for Medicine Culture Collections ( CMCC) and used for tracheitis vaccine production. Methods The test strain CMCC (B)29108 and the type strain DSM30007T were cultured on NA medium. Characteristics in morphology, physiology, biochemistry and fatty acid profile were compared between the two strains. Phylogenetic analysis was based on 16S rRNA and rpoB gene sequences, together with the DNA-DNA hybridization assay. Results A Comparative analysis of a partial sequence of the 16S rRNA gene sequence revealed that the CMCC( B) 29108 strain was closed to the Acinetobacter species and showed the highest similarity with the type strain Acinetobacter baumannii DSM30007T. Moreover, the CMCC(B)29108 strain was highly similar to type strain DSM30007T in morphology, physiology, biochemistry and fatty acid profile. On the phylogenetic tree based on 16S rRNA and rpoB gene of all Acinetobacter members, the CMCC(B)29108 strain steadily clustered into one independent branch only with the DSM30007 T strain with a DNA-DNA hybridization value of 100%. Conclusion The CMCC(B)29108 strain that is one of the strains used for the production of tracheitis vac-cine should be assigned to the species of Acinetobacter baumannii based on its phenotypic, phylogenetic and chemotaxonomic characteristics.
2.Analysis of chemical constituents of anti-myocardial ischemia fraction of Corydalis yanhusuo.
China Journal of Chinese Materia Medica 2008;33(14):1717-1719
OBJECTIVETo establish the HPLC fingerprint of anti-myocardial ischemia fraction of Corydalis yanhusuo, and to investigate its chemical constituents by LC-MS-MS.
METHODThe separation was performed at 30 degrees C on a Dikma Diamonsil C18 column (4.6 mm x 250 mm, 5 microm) by isocratic elution with acetonitrile-water (23:77) (containing 0.8% acetic acid and 0.2% triethylamine) as the mobile phase at a flow rate of 1.0 mL x min(-1), with UV detection at 335 nm.
RESULTA satisfactory fingerprint was obtained by HPLC method, 12 common peaks were selected as the fingerprint peaks, and 8 among them were identified according to the chromatographic behavior and mass spectroscopic data by comparison with those of authentic standard.
CONCLUSIONThis method is simple, sensitive and reliable, can be used to investigate the chemical constituents of anti-myocardial ischemia fraction of C. yanhusuo and control the quality.
Chromatography, High Pressure Liquid ; Corydalis ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Tandem Mass Spectrometry
3.A study on awareness of sexually-transmitted disease/acquired immunodeficiency syndrome and related factors among migrant workers in Hangzhou City, Zhejiang Province of China.
Xu-Jun YE ; Wei-Xing SHI ; Lu LI
Chinese Journal of Preventive Medicine 2004;38(5):316-320
OBJECTIVETo understand awareness on transmission routes of sexually transmitted diseases and acquired immunodeficiency syndrome (STD/AIDS) among migrant workers in Hangzhou City, Zhejiang Province.
METHODSA cross-sectional study was conducted in migrant workers in Hangzhou with self-administered anonymous questionnaire to collect their demographic information and awareness on STD/AIDS. All the data were analyzed by SPSS 11.0 software.
RESULTSA total of 3 001 subjects were interviewed. Most of them have already had some knowledge about STD/AIDS, but not complete. There were 556 (18.8%) migrant workers did not understand that condom could prevent from STD, and 759 did not know if it could do. There were 357 (11.9%) workers did not know AIDS could be prevented, and 746 (24.9%) thought that AIDS could be cured. There were 637 workers did not know that correct use of condom could reduce occurrence of AIDS, and 725 of them thought AIDS could be infected by hands-shaking and hugging with patients of AIDS. There existed statistically significant difference in awareness on STD/AIDS between men and women, workers with varied marital status and education levels.
CONCLUSIONSAwareness on STD/AIDS in migrant workers was smattering, allowing of not optimistic. Community-based health education on knowledge about STD/AIDS should be strengthened among high-risk migrant workers with varied channels to improve their awareness.
Acquired Immunodeficiency Syndrome ; psychology ; transmission ; Adult ; China ; epidemiology ; Condoms ; Cross-Sectional Studies ; Female ; Health Knowledge, Attitudes, Practice ; Humans ; Male ; Risk-Taking ; Sex Education ; Sexually Transmitted Diseases ; psychology ; transmission ; Surveys and Questionnaires ; Travel
4.Non-fusion soluble expression of broad-spectrum antivirus protein in Escherichia coli by translational-coupling with SUMO
Lingyue XING ; Dejian XIE ; Bingyu YE ; Zhang ZHANG ; Ping LI ; Wenlong SHEN ; Minglei SHI ; Yan ZHANG ; Zhihu ZHAO
Military Medical Sciences 2015;(8):597-601
Objective To design and construct a new non-fusion soluble expression vector pTIG-mSUMO(small ubiq-uitin-related modifier) using the widely used solubility promoting protein SUMO and based on the translational coupling phenomenon in order to enable the non-fusion soluble expression of the broad-spectrum antiviral protein RA in Escherichia coli by pTIG-mSUMO.Methods The smt3 gene coding for SUMO protein was cloned from yeast genome DNA by PCR. After directed-site silent mutation to eliminate the EcoRⅠsite, the mutant mSUMO was inserted into pET-22b to obtain the translational coupling expression vector pTIG-mSUMO.The RA was subject to PCR amplification and cloned into the pTIG-mSUMO to obtain the expression plasmid pTIG-mSUMO/RA which was supposed to direct the soluble expression of RA by the translational coupling with mSUMO.Results A translational coupling expression vector pTIG-mSUMO which could di-rect/drive the SUMO and heterogonous protein non-fusion expression simultaneously was designed and constructed.The Western blotting result indicated that pTIG-mSUMO could direct the high-level expression of RA, around 40%of which was soluble.Conclusion A translational coupling expression vector pTIG-mSUMO is obtained.After coupling with SUMO, RA is highly expressed in E.coli and both the expression level and solubility are greatly improved.pTIG-mSUMO might contrib-ute to soluble expression of other proteins.
5.A High-throughput and Quantitative Assay Based on Fluorescence Intensity for Detection of Apoptosis
Ling-Ling YE ; Hong LIU ; Xing-Mao LIU ; Shi-Chong LI ; Ben-Chuan WU ; Qi-Wei WANG ; Zhao-Lie CHEN ;
China Biotechnology 2006;0(06):-
Based on the different permeability of DNA-intercalant dyes YO-PRO-1(YP) and propidium iodide (PI) to the membrane of viable, apoptotic and necrotic cells, cell samples were stained with 4?mol/L YP and 4?g/ml PI for 10 min, and the fluorescence intensity of both YP and PI were measured by fluorometer at Ex/Em wavelength of 485/538nm and 530/590nm, respectively. The correlation between YP fluorescence intensity and the apoptotic cell number was confirmed by fluorescence microscope and linear regression(r=0.999,P
6.Case-control study on bone setting manipulation for the treatment of over degree II supination-eversion fractures of ankle joint.
Yue-Feng QI ; Fa-Lin CHEN ; Shu-Ren BAO ; Cheng-Huan LI ; Xing-Wei ZHAO ; Shi-Ming LIU ; Wen-Xue CHEN ; Ye LI ; Peng WANG
China Journal of Orthopaedics and Traumatology 2012;25(8):634-638
OBJECTIVETo explore therapeutic effects of bone setting manipulation for the treatment of over degree II supination-eversion fractures of ankle,and analyze manipulative reduction mechanism.
METHODSFrom 2005 to 2008, 95 patients with over degree II supination-eversion fractures of ankle were treated respectively by manipulation and operation. There were 43 cases [11 males and 32 females with an average age of (44.95 +/- 12.65) years] in manipulation group, and 2 cases were degree II, 11 cases were degree III, and 30 cases were degree IV. There were 52 cases [21 males and 31 females with an average age of (39.96 +/- 13.28) years] in operative group,and 6 cases were degree II, 18 cases were degree III, and 28 cases were degree IV. Bone setting manipulation and hard splint external fixation were applied to manipulative group. Operative reduction internal fixation was performed in operative group. X-ray was used to evaluate reduction of fracture before and after treatment, 2 months after treatment. Ankle joint function was evaluated according to Olerud-Molander scoring system after 6 months treatment.
RESULTSAll patients were followed up with good reduction. Three cases occurred wound complication in operative group, but not in manipulative group. In manipulation group, 19 cases got excellent results, 20 cases good and 4 cases fair; while in operative group, 30 cases got excellent results, 20 cases good and 2 cases poor. There were no significant differences in fracture reduction and ankle joint function recovery between two groups (P > 0.05). Efficacy of operative treatment was better than that of manipulative treatment at degree IV fracture (P < 0.05).
CONCLUSIONBone setting manipulation is a good method for treating supination-eversion ankle joint fractures, which has advantages of simple and safe operation, reliable efficacy. For ankle join fracture at degree IV, manipulative reduction should be adopted earlier, and operative treatment also necessary
Adult ; Ankle Injuries ; physiopathology ; therapy ; Ankle Joint ; physiology ; Case-Control Studies ; Female ; Fractures, Bone ; physiopathology ; therapy ; Humans ; Male ; Manipulation, Orthopedic ; methods ; Supination
8.Construction of Attenuated Influenza Virus Vaccines of Vaccines of-2006-2007
Penghui YANG ; Yi YE ; Xinfu SHI ; Deyan LUO ; Yu ZHANG ; Yan YAN ; Pengfei ZHANG ; Yueqiang DUAN ; Li XING ; Xiufan LIU ; Xiliang WANG
Progress in Biochemistry and Biophysics 2008;35(3):312-319
Six gene segments,PB1,PB2,PA,NP,M and NS,were fully synthesized which derived from the master donor virus (MDV),cold-adapted(ca),temperature sensitive(ts),live attenuated influenza virus strain A/Ann Arbor/6/60(MDV-A).Meanwhile,five amino acid substitutions (PB1-391E,58lG,661T,PB2-265S,NP-34G) were artificially altered by human intervention.HA and NA fragments derived from the 2006-2007 circulating strain A/New Caledonia/20/99 (H1N1).Eight fragments were ligated with modified pAD3000 for rescue plasmid construction.Eiight transcription/expression plasmids were named as pMDV-A-PB2,pMDV-A-PB1,pMDV-A-PA,pMDV-A-NP,pMDV-A-M,pMDV-A-NS,pMDV-A-HA,pMDV-A-NA,respectively.The COS-l cells were co-transfected with eight plasmids representing 6 internal viral backbone of the strain A/AA/6/60 and two plasmids containing the CDNA of the HA and NA segments of the strain A/New Caledonia/20/99 (H1N1),the results showed that cold-adapted,attenuated reassortant influenza A virus Was rescued successfully.Titers of a reassorted influenza A virus in embryonated chicken eggs mnged from 1:29to l:210.The rescue system of six intemal genes used as backbone opens the way for further research on gene function and neotype vaccine candidate of cold-adapted,live attenuated human influenza virus.
9.Study on quality specification of Rhizoma et Radix Valeriana Jatamansi.
Hong-ye DI ; Jin-li SHI ; Xing-li YAN ; Ren ZHAO ; Yong LIU ; Pei-gen XIAO
China Journal of Chinese Materia Medica 2007;32(22):2357-2359
OBJECTIVETo provide scientific basis for the utilization and development of Valeriana jatamansi by setting up the quality control specification of V. jatamansi.
METHODThe pharmacognostical methods were applied. The extract of V. jatamansi was examined. Moisture and ash were determined. And the bioactive constituents were analyzed by TLC and HPLC.
RESULTThe morphological and histological characters of V. jatamansi were observed. Content of total ash, acid-insoluble ash, and moisture of 15 samples from different habitats and times were determined. The qualitative and quantitative analysis of valtrate and acevaltrate by TLC and HPLC were preformed respectively.
CONCLUSIONThe established method can be used for the quality control of V. jatamans.
Chromatography, High Pressure Liquid ; Chromatography, Thin Layer ; Iridoids ; analysis ; Pharmacognosy ; standards ; Plant Roots ; anatomy & histology ; chemistry ; Plants, Medicinal ; anatomy & histology ; chemistry ; Quality Control ; Rhizome ; anatomy & histology ; chemistry ; Valerian ; anatomy & histology ; chemistry
10.Construction of Attenuated Influenza Virus Vaccines of 2006~2007
Penghui YANG ; Yi YE ; Xinfu SHI ; Deyan LUO ; Yu ZHANG ; Yan YAN ; Pengfei ZHANG ; Yueqiang DUAN ; Li XING ; Xiufan LIU ; Xiliang WANG
Progress in Biochemistry and Biophysics 2006;0(03):-
Six gene segments, PB1, PB2,PA, NP, M and NS, were fully synthesized which derived from the master donor virus(MDV), cold-adapted(ca),temperature sensitive(ts), live attenuated influenza virus strain A/Ann Arbor/6/60(MDV-A). Meanwhile, five amino acid substitutions (PB1-391E, 581G, 661T, PB2-265S, NP-34G) were artificially altered by human intervention. HA and NA fragments derived from the 2006~2007 circulating strain A/New Caledonia/20/99 (H1N1). Eight fragments were ligated with modified pAD3000 for rescue plasmid construction. Eight transcription/expression plasmids were named as pMDV-A-PB2, pMDV-A-PB1, pMDV-A-PA, pMDV-A-NP, pMDV-A-M, pMDV-A-NS, pMDV-A-HA, pMDV-A-NA, respectively. The COS-1 cells were co-transfected with eight plasmids representing 6 internal viral backbone of the strain A/AA/6/60 and two plasmids containing the cDNA of the HA and NA segments of the strain A/New Caledonia/20/99 (H1N1), the results showed that cold-adapted, attenuated reassortant influenza A virus was rescued successfully. Titers of a reassorted influenza A virus in embryonated chicken eggs ranged from 1∶29 to 1∶210. The rescue system of six internal genes used as backbone opens the way for further research on gene function and neotype vaccine candidate of cold-adapted, live attenuated human influenza virus.