2018年10期目次

  • Application of cell factory in production of a series of MMR vaccine

    FU Yong-qi;SONG Yan-mei;GAO Jing;ZHANG Ying-xin;LIU Hui;SUN Ya;SHEN Ya-nan;LV Miao;YANG Yun-kai;Beijing Bei Shengyan Biological Products Co.,Ltd.;

    Objective To culture measles,mumps and rubella viruses in cell factory and prepare the relevant monovalent and combined vaccines. Methods The bulks of measles,mumps and rubella viruses were prepared by culture of primary chick embryo cells and 2 BS cells in 40-layer cell factory,using those cultured in 15 L roller bottle as control,based on which the relevant monovalent and combined vaccines were prepared and determined for titer according to the requirements in Chinese Pharmacopoeia(Volume Ⅲ,2015 edition). Results The titers of bulks of measles,mumps and rubella viruses cultured in cell factory were 5. 2,6. 1 and 5. 7 lg CCID_(50)/m L respectively,which were significantly higher than those cultured in roller bottle and met the requirements in Chinese Pharmacopoeia(Volume Ⅲ,2015 edition). The yields of bulks prepared by a cell factory were equivalent to 4 ～ 8 times of those by culture in roller bottle. All the titers of monovalent and combined vaccines prepared by cell factory met the requirements in Chinese Pharmacopoeia(Volume Ⅲ,2015 edition),and decreased by not more than 1 lg in heat stability test. Conclusion The process for production of monovalent and combined measles,mumps and rubella vaccines by cell factory was stable,reproducible and controllable,which might used for large-scale production.

    2018 10 v.31 [Abstract][OnlineView][Download 265K]

  • Construction and immune effect of chitosan mediated pagC or nmpC DNA vaccine against Salmonella paratyphi A

    LIANG Hao-yu;REN Hui-mei;ZHANG Ying;DONG Si-guo;WANG Bin;ZENG Ming;Division of Intestinal Bacteria Vaccine,National Institutes for Food and Drug Control,Key Laboratory of the Ministry of Health for Research on Quality and Standardization of Biotech Product;

    Objective To construct a chitosan(CS)-mediated DNA vaccine expressing nmpC or pagC genes of Salmonella paratyphi A and investigate its immune response. Methods The sequences of nmpC and pagC genes were optimized according to the codon-preference of mammals,and cloned into eukaryotic expression vector provax to construct DNA vaccines provax-nmpC and provax-pagC respectively. CS nanoparticles loaded with DNA were prepared by complex coacervation method and analyzed for speciality. BHK cells were transfected in vitro with DNA vaccines provax-nmpC and provax-pagC and the corresponding CS nanoparticles respectively,and determined for expressions of NmpC or PagC by Western blot. Mice were immunized with the recombinant proteins by electric shock,and determined for serum antibody level and types by ELISA. Results Restriction analysis proved that DNA vaccines provax-nmpC and provax-pagC were constructed correctly. The vaccines were bound to CS nanoparticles effectively. The obtained vaccine-loaded nanoparticles were spherical in even size,of which the encapsulation rate was more than 90%. CS prevented DNA from digestion with DNaseⅠ. Both provax-nmpC and CS/provax-nmpC induced effective immune response in mice,while the response levels showed no significant difference(P > 0. 05). Likely,both provax-pagC and CS/provax-pagC induced effective immune response. However,the antibody level induced by provax-pagC was significantly higher than that by CS/provax-pagC(P < 0. 05). The IgG1 and IgG2 a levels induced by provax-nmpC and provax-pagC were higher than those by the corresponding CS-based vaccines(P < 0. 05). However,no significant difference was observed in IgG1 and IgG2 a levels.Conclusion Both DNA vaccines provax-nmpC and provax-pagC induced effective immune response in mice. CS nanoparticles failed to enhance the effect or alter the type of immune response to DNA vaccine while regulated the balance between Th1 and Th2 responses.

    2018 10 v.31 [Abstract][OnlineView][Download 382K]

  • Optimization of propagation conditions for cultivation of influenza A virus on adherent MDCK cells

    XI Li;LIU Xu-guang;LI Shuang;LI Ke-xin;LI Xue;ZOU Yu-ying;ZOU Yong;Changchun Institute of Biological Products Co.,Ltd.;

    Objective To optimize the condition for propagation of influenza A virus on adherent MDCK cells. Methods Three influenza A virus strains,H1N1 NYMC X-181,H1N1 NYMC X-275 and H3N2 NYMC X-263 B,were subcultured on adherent MDCK cells,of which the condition for propagation,including trypsin concentration(0. 5,1. 25,2. 5,3. 5 and 4. 5 μg/m L),MOI(0. 000 1,0. 001,0. 01,0. 1 and 1),temperature(33,34,35,36 and 37 ℃),adsorption time(0,1. 0,1. 5,2. 0,2. 5 and 3. 0 h),cultivation time(24,48,72,96 and 120 h),were optimized,using hemagglutination titer and median cell culture infective dose(CCID_(50)) as indicators. Results The optimal trypsin concentrations for propagation of H1N1 NYMC X-181,H1N1 NYMC X-275 and H3N2 NYMC X-263 B strains were 2. 5,1. 25 and 1. 25 μg/m L respectively,while all the optimal MOIs were 0. 01,and all the optimal culture temperature were 34 ℃. The optimal adsorption times of the three strains were 1. 5 ～ 2. 0,1. 5 and 1. 5 h respectively,while all the optimal cultivation times were 72 h. Conclusion The condition for propagation of three influenza A virus strains on adherent MDCK cells were preliminarily optimized,which laid a foundation of further development of relevant vaccines.

    2018 10 v.31 [Abstract][OnlineView][Download 299K]

  • Expression of human heat shock protein 10 in Pichia pastoris and purification of expressed product

    LI Xiao-ying;HOU Zeng-miao;DENG Han;LI Min;LI Zhe;YANG Xiao-lin;ZHAO Jin-li;Shaanxi Huikang Bio-tech CO.,Ltd;

    Objective To express human heat shock protein 10(Hsp10) in Pichia pastoris and purify the expressed product. Methods Total RNA was extracted from fresh human placenta,reversely transcribed to cDNA and used as a template for amplification of Hsp10 gene fragment by PCR,based on which recombinant plasmid p PIC9 K-hsp10 was constructed and transformed to P. pastoris GS115 by electroporation. Positive strain pPIC9 K/Hsp10/GS115 was obtained by histidine defect screening,and induced by methanol. The protein content of expressed Hsp10 was determined by SDSPAGE,while the specificity was analyzed by Western blot. Results Hsp10 gene with correct sequence and the strain for high expression of the gene were obtained. SDS-PAGE profile showed specific band with a relative molecular mass of 11 000.The expression level of Hsp10 was 400 mg/L,while the purity was more than 90%. Western blot showed good reactogenicity and specificity of Hsp10,which were consistent with those expected. Conclusion Hsp10 was successfully expressed in P. pastoris GS115,which laid a foundation of large-scale production and application of the protein.

    2018 10 v.31 [Abstract][OnlineView][Download 348K]

  • Interaction of host TRIM56 protein with Npro protein of classical swine fever virus and effect of TRIM56 on virus replication

    ZHANG En-yu;ZHU Yan;WANG Jing-han;LI Su;QIU Hua-Ji;SHI Dong-fang;College of Animal Medicine,Northeast Agricultural University;

    Objective To analyze the interaction between TRIM56 and N~(pro) protein of classical swine fever virus(CSFV)and investigate the effect of host TRIM56 on replication of the virus. Methods PK-15 cells in which the expression of TRIM56 was down-regulated by using si RNA were inoculated with CSFV. The effect of the down-regulation of TRIM56 expression on proliferation of CSFV was evaluated by Western blot and fluorescent quantitative RT-PCR. The interaction between TRIM56 and N~(pro)was confirmed by co-immunoprecipitation(Co-IP) assay and Western blot. The co-location of TRIM56 and N~(pro)was determined by laser confocal microscopy. Results Both the genomic copy number and titer of CSFV of down-regulation of TRIM56 expression increased significantly. TRIM56 and N~(pro)were interacted specifically and colocalized in the cytoplasm of HEK 293 T cells. Conclusion Host TRIM56 showed interaction with CSFV N~(pro)protein and negatively regulated the replication of CSFV.

    2018 10 v.31 [Abstract][OnlineView][Download 299K]

  • Stability of pneumococcal polysaccharide of 13 serotypes

    REN Zhen-yun;LIU Qian;ZHANG Yi;CHEN Xiao-hang;LIU Ai-ping;WANG Xi;XIONG Ming-yu;SHEN Rong;First Department of Research,Lanzhou Institute of Biological Products Co.,Ltd.,Center for Gansu Provincial Vaccine Engineering Research;

    Objective To investigate the stability of pneumococcal polysaccharide(Pn Ps) of 13 serotypes stored at-60 ℃ for 60 months. Methods Pn Ps of serotypes 1,3,4,5,6 A,6 B,7 F,9 V,14,18 C,19 A,19 F and 23 F,three batches for each,were stored at-60 ℃,from which samples were taken every 6 months and determined for composition content,distribution coefficient(K_D)value,molecular weight distribution(HPSEC-MALLS/RI)and antigen activity. The total time length for determination was 60 months. Results The composition content and K_Dvalue of pneumococcal polysaccharide stored at-60 ℃ for 60 months met the relevant quality standards. The results of HPSECMALLS/RI showed no significant change. The Pn Ps showed high antigen activity. Conclusion The pneumococcal polysaccharide of 13 serotypes showed high stability after storage at -60 ℃ for 36 months.

    2018 10 v.31 [Abstract][OnlineView][Download 506K]

  • Target gene prediction and bioinformatics analysis of MiR-124a in dairy cows

    YANG Zhuo-ni-na;WANG Xiu-wei;GUO Li;ZHANG Wei-hong;YANG Huan-min;SHEN Bing-lei;LIU Sheng-jun;College of Animal Science and Veterinary Medicine,Heilongjiang Bayi Agricultural University;

    Objective To predict the target gene of miR-124 a(bta-miR-124 a) of dairy cows for enrichment analysis of biological function,regulation mechanism and signal transduction pathways,and provide a theoretical basis for the functional verification of miR-124 a in milk quality regulation in dairy cows. Methods The precursor and mature sequences of bta-miR-124 a and candidate target genes were predicted by using miRBase,miRanda and Targetscan online analysis systems. GO function annotation and signal pathway enrichment analysis were performed on the confirmed target genes. Results The premature and mature sequences of bta-miR-124 a were highly conserved among species,of which the candidate target genes were mainly enriched in ten biological processes such as biological regulation,cell growth and metabolism. The KEGG signaling pathway showed that the candidate target genes were mainly enriched in the pathways of cancer,mitogen-activated protein kinase(MAPK),AMP-activated protein(AMPK),cyclic adenosine monophosphate(c AMP),fatty acid degradation and fatty acid biosynthesis. Conclusion The candidate genes of bta-miR-124 a play important roles in onset of cancer,AMPK signal transduction and fatty acid metabolism. Fatty acid metabolism is the main metabolic pathway in which bta-miR-124 a may participate. It provides a reference for research on the functional mechanism of bta-miR-124 a in regulation of butterfat metabolism in dairy cows.

    2018 10 v.31 [Abstract][OnlineView][Download 281K]

  • Gene variation of measles virus nucleoprotein in Jilin Province,China during 2001～2014

    HUANG Biao;SHAN Yuan-chun;ZHOU Jian-hui;WANG Shuang;TIAN Xin;YANG Yao;WEI Lei-lei;Jilin Provincial Center for Disease Control and Prevention;

    Objective To analyze the gene variation of measles virus nucleoprotein(N) in Jilin Province,China from2001 to 2014. Methods The N gene sequence of measles virus was obtained by virus isolation, extraction and amplification of RNA and sequencing,of which the variation was analyzed by using Bio Edit and Mega software. Results The incidences of measles were high in 2005,2006,2009 and 2014. The variation rates of nucleotide(nt) and amino acid(aa) of epidemic strains of measles virus were 0. 59% ～ 1. 85% and 1. 00% ～ 2. 67% respectively. The mutation of L →I appeared at aa100 which might be the specific site in Jilin Province in various years. However,the back mutation of S → G appeared at aa82 in 2009 and last to 2014,which was localized in the same strain with those at aa 47 and aa122. The mutations at aa63(R→I),aa69(S→T),aa77(R→L) and aa82(R→I) appeared in 2013 and 2014.The missense mutation rates were the highest in 2009 and 2010. The mutations at aa47,aa82 and aa122 were epidemic for many years,of which the entropies were higher than those at other sites. Conclusion Both the variation rates of nt and aa of measles virus strains in Jilin Province during 2001 ～ 2014 increased year by year. The amino acid variation in the key sites of should be monitored continuously to provide an early warning for epidemic of measles in combination with epidemiological data.

    2018 10 v.31 [Abstract][OnlineView][Download 383K]

  • Effects of coexpression of molecular chaperones on expression and bioactivity of recombinant bovine fibroblast growth factor-2 in Escherichia coli

    SUN Dan;WANG Li-jie;XU Xiao-hui;HE Wei;School of Health and Nutrition,He University;

    Objective To investigate the effects of coexpression of molecular chaperones on expression and bioactivity of recombinant bovine fibroblast growth factor-2(rb FGF-2)in Escherichia coli. Methods The nucleotide sequence of a fulllength rbFGF-2 was designed and synthesized according to the codon usage preferences in E. coli,based on which an expression vector pET-15b-rbFGF-2 was constructed. The pGro7 plasmid encoding the chaperones GroES-GroEL and the vector pET-15b-rbFGF-2 were transformed successively into E. coli for coexpression. The expressed product was identified by Western blot and purified by Ni-Agarose affinity chromatography,of which the bioactivity was determined by MTS assay.Results Restriction enzyme digestion analysis and DNA sequencing confirmed that the sequence of rbFGF-2 gene was identical to the designed sequence and the target gene was inserted into Nde Ⅰand Bam HⅠsites of the expression vector.The expressed rbFGF-2 protein,with a relative molecular mass of approximately 21 000,was produced mainly in the form of inclusion bodies representing 37. 8% of total bacterial protein. The rbFGF-2 protein showed specific binding to monoclonal antibody against bFGF-2. More than 95% purity was confirmed by SDS-PAGE after purification. The expression level of rbFGF-2 in E. coli containing co-expression system was 2. 28 times as high as that in E. coli containing only the expression vector. The rbFGF-2 protein at a concentration of 6. 25 ng/ml significantly promoted the cell proliferation.Conclusion The co-expression of chaperones GroES-GroEL significantly increased the expression level and bioactivity of rbFGF-2 protein.

    2018 10 v.31 [Abstract][OnlineView][Download 257K]

  • Effect of XiaSangju on influenza A (H1N1) virus and relevant mechanism

    YU Si-ming;LI Yan-hua;JIANG Yi;WANG Lei;The First Hospital of Jilin University;

    Objective To investigate the effect of XiaSangju,a preparation of traditional Chinese drug,on influenza A(H1N1)virus as well as the relevant mechanism. Methods The median effective concentration(TC_(50))to MDCK cells and median inhibitory concentration(IC_(50))of influenza A(H1N1)virus of XiaSangju were determined by MTT assay,based on which the selection index(SI)was calculated. The median tissue culture infective dose(TCID_(50))of the virus to MDCK was determined by Reed-Muench method. The effect of XiaSangju on influenza A(H1N1)virus was investigated by plaque reduction neutralization test,while that on the location of NP by fluorescent staining. The expression of NF-κB pathway-associated proteins in the cells infected with influenza A(H1N1) virus was determined by Western blot. Results The TC_(50),IC_(50),SI and TCID_(50) were 4. 98 mg/m L,2. 063 mg/m L,2. 414 and 0. 044 668 respectively. XiaSangju inhibited the plaque formation of influenza A(H1N1),the export of NP from nuclei as well as the phosphorylation of NF-κB pathway-associated proteins Iκκα,Iκκβ,NF-κBp50 and NF-κBp65. Conclusion XiaSangju showed significantly inhibitory effect on influenza A(H1N1)virus by a possible mechanism of inhibiting the phosphorylation NF-κB pathwayassociated proteins.

    2018 10 v.31 [Abstract][OnlineView][Download 322K]

  • Prokaryotic expression of Mycobacterium tuberculosis PPE36-p27 protein and its application in diagnosis of tuberculosis

    GUO Rui;GUO Yu-feng;JU Jun;HUANG Huan-yuan;WEI Zhen-hong;Gansu Provincial Hospital;

    Objective To express the PPE36-p27 protein of standard strain of Mycobacterium tuberculosis in prokaryotic cells and use the expressed product for determination of M. tuberculosis antigen-specific cellular immunity. Methods PPE36 gene was amplified by PCR using the genomic DNA of M. tuberculosis H37 RV as a template and inserted into expression vector p Easy. The constructed recombinant plasmid PEasy-PPE36 was transformed to E. coli BL21(DE3),and the expressed PPE36-p27 protein was identified by SDS-PAGE,purified by Ni-NTA chromatography and used as a specific stimulator to develop a novel ELISPOT-IL-12 assay for T cells. Results Restriction analysis and sequencing proved that recombinant plasmid p Easy-PPE36 was constructed correctly. The expressed PPE36-p27 protein,with a relative molecular mass of about 30 000,reached a concentration of 14. 69 μg/m L after purification,and stimulated the production of IL-12 in ELISPOT-IL-12 assay. The produced IL-12 showed specific binding to the coated antibody against IL-12,which formed obvious spots. Conclusion PPE36-p27 protein was successfully expressed in prokaryotic cells,with which an ELISPOT-IL-12 assay was developed and used for the early diagnosis of tuberculosis.

    2018 10 v.31 [Abstract][OnlineView][Download 362K]

  • Surveillance of adverse events following immunization in Hongshan District,Wuhan City,Hubei Province,China during 2010～2016

    XU Dan-dan;ZHU Ming-qing;LIU Zhi-sheng;CHEN Jie;ZHAO Ming-jiang;Hongshan District Center for Disease Prevention and Control,Wuhan;

    Objective To analyze the characteristics and influencing factors of adverse events following immunization(AEFI)in Hongshan District,Wuhan City,Hubei Province,China from 2010 to 2016 so as to improve the sensitivity of surveillance. Methods AEFI data and immunization information from 2010 to 2016 were collected through the National Immunization Program Information Management System and Children′s vaccination reports. The epidemiology of relevant indicators was analyzed by descriptively epidemiologic methodology. Results A total of 385 cases of AEFI were reported from 2010 to 2016 in Hongshan District of Wuhan City,with an annual reported rate of 17. 85/100 000 doses,including295 cases of general reactions(76. 62%). The most of symptoms were fever,swelling and indurations. There were 72 cases of abnormal reactions(18. 70%),of which the main manifestations were allergic rashes(27. 78%),papules(40. 28%)and urticaria(25%). Three cases of severe abnormal reactions(4. 17%)were observed,including 2 cases of allergic purpura and one case of thermal convulsion. No vaccination accidence occurred. A portion of 78. 18% of the cases appeared in the children at ages of less than 2 years. The most of the cases were reported from April to September every year. The vaccines of class Ⅰ associated with the AEFI were diphtheria and tetanus combined vaccine(DP),Japanese encephalitis(JE)vaccine and measles-containing vaccines(40. 43/100 000 doses),while those of classⅡwere pertussis,diphtheria,tetanus and Haemophilus influenzae type b(Hib) combined vaccine and 23-valent pneumococcal polysaccharide,diphtheria,tetanus and acellular pertussis,inactivated poliomyelitis and Haemophilus influenza type b combined vaccine. The reported rate of AEFI caused by vaccines of class Ⅰ was 18. 90/100 000 doses,which was significantly higher than that of class Ⅱ(7. 09/100 000 doses)(P < 0. 01). Conclusion The AEFI surveillance system in Hongshan District is running well and the reported rate of AEFI is increasing. However,the sensitivity of monitoring should be further strengthened.

    2018 10 v.31 [Abstract][OnlineView][Download 224K]

  • Seroprevalence of hepatitis B in populations at ages of 1～59 years in Changchun City,Jilin Province,China in 2006 and 2016

    TAO Yu-hui;YAN Li;WANG Chong;XU Hong-qin;ZHANG Jun-peng;YU Jian-xing;Changchun Municipal Center for Disease Control and Prevention;

    Objective To compare the seroprevalence of hepatitis B in populations at ages of 1 ～ 59 years in Changchun City,Jilin Province,China in 2006 and 2016. Methods A multi-stage cluster random sampling method was used for analysis of hepatitis B surface antigen(HBsAg) carriage rate,hepatitis B surface antibody(HBsAb) positive rate and coverage of hepatitis B vaccine(Hep B) in populations at ages of 1 ～ 59 years in Dehui County and Nanguan District of Changchun City. Results The HBsAg carriage rates in 932 subjects in 2006 and 1 097 subjects in 2016 were 3. 22% and 1. 64%,while the HBsAb positive rates were 51. 07% and 52. 51%,and the HepB coverage rates were 58. 15% and 67. 73%,respectively,which showed significant difference in the subjects at various age stages(P < 0. 05),while showed no significant difference in male and female subjects(P > 0. 05). Both HBsAb positive rate and HepB coverage rate showed significant difference in urban and in rural areas(P < 0. 05). However,the HBsAg carrier rate showed significant difference in urban and rural areas in 2006(P < 0. 05),while showed no significant difference in 2016(P >0. 05). Conclusion During the past ten years,the HepB coverage rate in Changchun increased,while the HBsAg carriage rate decreased,and the HBsAb positive rate showed no significant change.

    2018 10 v.31 [Abstract][OnlineView][Download 158K]

  • Effects of controlled released BMP-2 and VEGF from PELA microcapsule-based scaffolds on Wnt/β-catenin pathway for differentiation of bone mesenchymal stem cells to osteoblasts

    REN Qian-gui;WANG Zhao;ZHANG Kun;Second Affiliated Hospital of Inner Mongolia Medical University;

    Objective To prepare the VEGF-encapsulated microcapsules with external adhesion of recombinant human bone morphogenetic protein 2(rh BMP-2)and investigate their effect on the Wnt/β-catenin pathway for differentiation of bone mesenchymal stem cells(BMSCs) to osteoblasts. Methods The VEGF-encapsulated microcapsules with external adhesion of rh BMP-2 were prepared with polylactide-poly-ethylene glycol-polylactide(PELA), based on which microcapsule scaffolds were prepared. The PELA microscapsule scaffolds containing BMP-2(BMP-2/PELA),VEGF(PELA/VEGF) and BMP-2 + VEGF(BMP-2/PELA/VEGF) were observed in this study,using the empty microscapsule scaffolds as control. The concentrations of rh BMP-2 and VEGF released by microcapsule scaffolds in PBS on 1,2,4,8,12,16,22,28,35,42 d after preparation were determined by ELISA. The activities of BMSCs cultured in the microscapsule scaffolds for 3,7 and 14 d were determined by MTT assay,while the expressions of Wnt/β-catenin pathway and alkaline phosphatase(ALP)by Western blot. Results About 60% of BMP-2 and about 32% of VEGF were released into PBS by the microscapsule scaffolds on day 2 after preparation. However,the activities of BMSCs in four groups after culture in microscapsule scaffolds for 3,7 and 14 d showed no significant difference(P > 0. 05). The expression levels of Wnt,β-catenin and ALP in BMP-2/PELA/VEGF group on day 14 after preparation were significantly higher than those on days 3 and 7(P < 0. 05),while were significantly higher on day 7 than on day 3(P <0. 05). Conclusion BMP-2/PELA/VEGF microscapsule scaffolds were prepared successfully,which showed no cytotoxicity to BMSCs and promoted the differentiation of BMSCs to osteoblasts.

    2018 10 v.31 [Abstract][OnlineView][Download 319K]

  • Effect of BCG vaccination in infants in two counties/districts of Huangshan City,Anhui Province,China

    ZHANG Xiang-yang;LUO Wei-qun;YANG Jing;The Center for Disease Control and Prevention of Huangshan City;

    Objective To analyze the effect of BCG vaccination in Qimen County and Huizhou District of Huangshan City,Anhui Province,China. Methods BCG-PPD test was performed on the infants in Qimen County and Huizhou District of Huangshan City 12 weeks after BCG vaccination,of which the result was analyzed statistically. Results The scar formation rate and BCG-PPD positive rate of infants in two counties/districts in Huangshan City were 99. 03% and89. 37% respectively. The positive rate of BCG-PPD was significantly higher in Qimen(93. 46%) than in Huizhou(85. 00%),and in urban(95. 19%)than in rural area(83. 50%)(each P < 0. 01). However,the BCG-PPD positive rate in male infants(89. 00%)showed no significant difference with that in female ones(89. 72%)(P > 0. 05). Both the average scar diameters in Qimen and Huizhou were 4. 32 mm. The BCG-PPD positive rate in infants with average scar diameters of not less than 3 mm was 92. 49%,which was significantly higher than that with average scar diameters of less than 3 mm(73. 53%)(P < 0. 01). However,the BCG-PPD positive rate of infants vaccinated in the obstetric wards of hospitals of not less than the second-class showed no significant difference with that in vaccination clinics(P > 0. 05).Conclusion The qualities of BCG vaccination in the two counties/districts of Huangshan City were satisfactory in general. However,there was an imbalance between urban and rural areas. The average scar diameter might be served as one of the reference for BCG vaccination quality.

    2018 10 v.31 [Abstract][OnlineView][Download 170K]

  • Safety of vaccination in children with congenital heart disease

    ZHENG Dong-yi;LIU Zhao-qiu;MA Song;ZHANG Ai-ling;ZHANG Wen-jing;SHI Nian-min;The First Hospital Affiliated to Tsinghua University;

    Objective To investigate the safety of vaccination in children with congenital heart disease. Methods A total of 69 children with congenital heart disease who received vaccination in The First Hospital Affiliated to Tsinghua University during July 2013 to July 2017 were subjected to the active monitoring of safety after vaccination,using those from the same communities,of the same genders,who received the same vaccines on the same dates as control. Results In the 69 children including 15 ones healed after surgery and 54 ones self-healed without surgery or unhealed,who were vaccinated at different times,only 8 cases of adverse events appeared,including 5 cases unrelated to the vaccination,all of which recovered to normal within 24 h. The incidence of adverse reactions showed no significant difference with that in control group(P > 0. 05). Conclusion The incidence of adverse reaction in children with congenital heart disease with normal heart function and body health were low,indicating a high safety.

    2018 10 v.31 [Abstract][OnlineView][Download 148K]

  • Optimization of preparation process for working virus seeds of live attenuated mumps vaccine

    LIU Hui;SUN Ya;WANG Gang;SONG Yan-mei;LU Meng-xi;WANG Dong-yan;FU Yong-qi;Beijing Bei Shengyan Biological Products Co.,Ltd.;

    Objective To control the basal titer and stability of mumps virus in live attenuated measles,mumps and rubella(MMR) combined vaccine by optimizing the preparation process of the mumps virus working seeds. Methods Fully-automated cell counter was used to improve the precision of chick-embryo cell counting. MOI was determined and the mumps working seed lot was prepared at the optimal MOI. The harvest time was optimized by plotting the complete virus growth curve to optimize the preparation process of the mumps virus working seed lot. Three consecutive batches of mumps virus bulk were prepared with the optimized virus seeds and determined for titer,with which the prepared live attenuated MMR combined vaccine were determined for basal titer of mumps virus as well as the titer after storage at(37 ± 1)℃ for 7 d. Results The titer of mumps virus bulk prepared with the optimized virus seeds was in the range of 6. 5 ～ 7. 0 lg CCID_(50)/m L,which was 0. 5 ～ 0. 7 log higher than that prepared with the virus seeds before optimization. However,the basal titer of mumps virus in MMR prepared with the optimized bulk was 5. 2 ～ 5. 8 lg CCID_(50)/m L,while that after storage at(37 ± 1) ℃ for 7 d was 4. 8 ～ 5. 5 lg CCID_(50)/m L. The ratios of measles virus∶mumps virus ∶rubella virus in 80% of MMR combined vaccine reached 1 ∶ > 20 ∶ 1. Conclusion Both the basal titer and stability of mumps virus in MMR combined vaccine prepared with the optimized working seeds of mumps virus were improved significantly.

    2018 10 v.31 [Abstract][OnlineView][Download 267K]

  • Identification of human diploid cell KMB17 strain for production of vaccines for human use

    SONG Cai-hua;REN Fang-fang;LIU Jie;DUAN Nan;QIAN Xing-li;LI Ya-xian;YANG Xiao-lei;HONG Chao;Institute of Medical Biology,Peking Union Medical College,Chinese Academy of Medical Science,Laboratory of Quality Control & Yunnan Provincial Key Laboratory For Development of Vaccines against Major Infections Disease;

    Objective To identify the human diploid cell KMB17 strain developed by the Institute of Medical Biology,Peking Union Medical College,Chinese Academy of Medical Science for production of vaccine for human use. Methods Chromosome karyotype and short tandem repeat(STR) genotyping were used. KMB17 cells for vaccine production were treated with colchicine to accumulate metaphase chromosomes and then release chromosomes at low permeability. The chromosomes were stained with Giemsa dye and observed for structure,of which the number in 100 cells were recorded precisely by microscopy. Twenty STR loci from human cells were subjected to DNA typing,and the obtained STR profiles were compared with those in ATCC and DSMZ data banks. Results The genome of sample cells showed normal structure under high power microscope,without deletion or mutation. Of the 100 cells counted,84 with 46 chromosomes were observed. However, no chromatid, chromosome fragmentation or chromosome structural aberration were observed.Fourteen hypodiploid and two hyperdiploid cells were found. STR profiles were unambiguous and no triplicate peak was observed. No 100% matched cell lines were found in ATCC and DSMZ data banks. Conclusion The established human diploid cell KMB17 strain for vaccine production was identified as correct,of which the structure and number of chromosomes were normal,and no contamination or cross-contamination was found.

    2018 10 v.31 [Abstract][OnlineView][Download 345K]

  • Optimization of refolding process for soluble tumor necrosis factorα receptor Ⅰ inclusion body by Design of Experiment

    QIN Jiao-rong;CAI Feng;ZHAO Zhao;DAI Yun-jian;LI Chun-yang;Chengdu Institute of Biological Products Co.,Ltd;

    Objective To optimize the key parameters in refolding process of recombinant soluble human tumor necrosis factor a receptorⅠ(sTNFα RⅠ)by Design of Experiment(DoE). Methods Two key technological parameters in refolding process,free sulfhydryl concentration(C-SH) and protein concentration(C-Pro),were optimized by using the Central Composite Quadratic Model established by Central Composite Design in DoE software. Two response values,the yield of sTNFαRⅠof each gram of wet bacteria and refolding volume(V_(-10g))for preparation of 10 g of sTNFαRⅠ,were set. The experimental data of model were evaluated by using DoE software. Under the premise of the highest yield of sTNFα RⅠand the minimum V_(-10g),the optimal refolding parameter values were predicted by response surface of quadratic model.The optimization results were validated by two scaled-up experiments. Results The center points of response surface of quadratic model established by 14 groups of tests of two Blocks at 2 factors and 5 levels showed high reproducibility,and the test points were showed a normal distribution. In regression analysis of response values for quadratic response surface model fitting,the lack of fit P values using sTNFα RⅠand V_(-10g)as response values were more than 0. 05,indicating that the lack of fit was insignificant. The predicted values are closed to the actual values,indicating high goodness of fit.Adeq Precisior values were more than 4,indicating that the model was effective and could be used to navigate the design space of optimization of refolding process parameters. The results showed that the yield of sTNFα RⅠwas 16. 38 mg/g and the V_(-10g) was 36. 73 L when the C_(-Pro) was 0. 48 mg/m L and the C_(-SH) was 8. 69 mmol/L in refolding process and the highest desirability of model was 0. 643. However,in validation by two scale-up experiments,the yields of sTNFα R Ⅰwere 11. 34 and 9. 72 mg/g respectively,which were higher than that before optimization(5. 1 mg/g in average).Conclusion The Central Composite Model may be used for screening of the parameters in refolding process of sTNFα RⅠ,which provides reliable data support for the subsequent scale-up of process.

    2018 10 v.31 [Abstract][OnlineView][Download 562K]

  • Progress in research on inactivated influenza vaccine

    NIAN Xuan-xuan;YANG Xiao-ming;ZHANG Jia-you;Wuhan Institute of Biological Products Co.,Ltd.;

    Influenza virus has the characteristics of antigenic drift and antigenic shift,which causes epidemic easily. At present,the most popular influenza vaccine in China is the inactivated influenza vaccine(IIV) which mainly against influenza A and influenza B virus. The preparation of broad-spectrum influenza vaccine is a challenge in vaccine development. Virologists and immunologists have promoted the understanding of immune basis of influenza virus and made great breakthroughs in research on inactivated vaccine through genetic engineering technology,virus cultivation and the establishment of animal models. This paper summarizes the mechanism of influenza epidemic and the latest progress in research on inactivated influenza vaccine.

    2018 10 v.31 [Abstract][OnlineView][Download 216K]

  • Advances in study of acute phase protein as a marker for diagnosis of animal stress

    YUAN Jian-bin;ZHENG Shu-sen;YANG Huan-min;GUO jing-ru;College of Animal Science and Veterinary Medicine,Heilongjiang Bayi Agricultural University;

    Stimulation by various stressors may induce non-specific defense response,which is known as acute phase response. Acute phase protein is mainly synthesized by hepatocytes,which may alter the expression of inflammatory responses to tissue damage,infection and stress,and is closely related to the static changes of environment and the antistress ability in stress response of animals. Therefore,it is necessary to explore the changes of acute phase protein in livestock and poultry stress-related diseases. In this paper,the classification of several major acute phase proteins and the change and detection of acute phase protein expression in animal stress are reviewed.

    2018 10 v.31 [Abstract][OnlineView][Download 156K]


@2012 Editorial Department of "Chinese Journal of Biologicals"