2018年04期目次

  • Adaptability of hepatitis A virus YN5D strain to human diploid cell line W2

    XU Dao-jun;LU Wei;QIAN Wen;DUAN Qing-tang;MA Bo;LIU Xin;WALVAX Biotechnology Co., Ltd.;

    Objective To investigate the subculture adaptability of hepatitis A virus(HAV) YN5D strain to human diploid cell line Walvax-2(W2)and determine the optimal condition for culture of the strain on W2 cells. Methods The P23 of HAV YN5 strain was inoculated into a T225 cell culture flask and subcultured to P30. The YN5D strain of various passages were determined for antigen titer and infectious titer. HAV YN5D strain P29 was inoculated to W2 cells at various MOIs to optimize the MOI, then inoculated to the cells at optimal MOI to optimize the time for harvest. Three-tier seed bank of HAV YN5D strain was established and subjected to control tests. Results The antigen and infectious titer of HAV YN5 strain were more than 10 240 EU/mL and more than 7. 00 lgCCID_(50)/mL after subculture for 30 passages respectively. Both the titers reached the maximum w hen the MOI was 0. 5. After culture for 24 ~ 28 d, the antigen and infectious titer of HAV YN5D strain were 2 560 EU/mL and 7. 24 lgCCID_(50)/mL respectively. All the results of control tests on the established three-tier virus seed bank met the requirements in Chinese Pharmacopoeia(Volume Ⅲ, 2015 edition). Conclusion The adaptability of hepatitis A virus YN5D strain on human diploid cell line W2 was determined,and the culture condition in cell factory was optimized, which laid a foundation of preparation of samples of HA vaccine used for clinical trial with human diploid cells.

    2018 04 v.31 [Abstract][OnlineView][Download 152K]

  • Stability of groups A and C meningococcal polysaccharide-CRM197 conjugate vaccine

    MA Hong-chu;ZHOU Li-bao;ZHANG Jing-chun;LIU Xiao-lei;WANG Zhen;LIU Feng;CHU Zi-qing;WANG Zhen-ye;SUN Shu-xue;Beijing ChengdaTianhe Biotechnology Co.,Ltd.;

    Objective To analyze the changes of main quality indexes of bulk and final product of groups A and C meningococcal polysaccharide conjugate vaccine using CRM 197,an avirulent mutant of diphtheria toxin,as carrier protein under various conditions for storage,and provide a basis for determination of condition for storage and validity period of the vaccine. Methods The bulks of groups A and C conjugate were stored separately at 2 ~ 8 ℃ for 3,6 and 8 months,and at 20 ~ 25 ℃ for 4 and 6 weeks,while the final product was stored at 2 ~ 8 ℃ for 3,6,12,18 and 24 months,at 20 ~ 25 ℃ for 3 and 6 months,and at 35 ~ 37 ℃ for 3 and 4 weeks,of which the main quality indexes were determined. Results All the quality indexes of bulk of group C met the relevant requirements after storage at 2 ~ 8 ℃ for 8 months and at 20 ~ 25 ℃ for 6 weeks,while those of group A at 2 ~ 8 ℃ for 8 months and at 20 ~ 25 ℃ for 3 weeks.However,all the quality indexes of final product met the relevant requirements after storage at 2 ~ 8 ℃ for 24 months,at 20 ~ 25 ℃ for 6 months and at 35 ~ 37 ℃ for 4 weeks. Conclusion Both the bulk and final product of groups A and C meningococcal polysaccharide conjugate vaccine using CRM 197 as carrier showed high stability.

    2018 04 v.31 [Abstract][OnlineView][Download 221K]

  • Comparison and analysis of structure and function of serine proteases from Trichinella spiralis in two development stages

    ZHAI Cheng-cheng;CHEN Jia-xu;CHEN Shao-hong;WU Xiu-ping;National Institute of Parasitic Diseases,Chinese Center for Disease Control and Prevention;WHO Collaborating Centre for Tropical Diseases;National Center for International Research on Tropical Diseases,Ministry of Science and Technology;Key Laboratory of Parasite and Vector Biology,Ministry of Health;

    Objective To compare and analyze the structure and function of serine proteases from intestinal adult worm(SP1)and newborn larvae(SP2)of Trichinella spiralis. Methods By using online bioinformatics websites and software,such as NCBI,EMBL,MEGA,ExPASY,TMHMM,SignalP,PROSITE,Coils and SYFPEITHI,the homology and phylogenesis of serine protease genes Zh68 and T668 from SP1 and SP2 of T. spiralis were analyzed,and the basic physiochemical property,transmembrane domain,motif,kringle domain,subcellular localization as well as secondary and tertiary structures of SP1 and SP2 were predicted. Results Zh68 and T668 were specific genes of T. spiralis,of which the homology of nucleotides was 48%,while that of amino acids was 27. 5%. Both SP1 and SP2 were unstable hydrophilic proteins,at which the N-terminuses there were secreted proteins with a signal peptide. The secondary and tertiary structures of the two proteins were similar,and the optimal antigen epitopes of B and T cells were predicted.However,SP2 was a membrane protein with a transmembrane helical structure. Conclusion SP1 is a non-transmembrane protein which may participate in the growth and reproduction of body parasitism in the intestine,while SP2 is a transmembrane protein which may participate in the invasion and migration of body parasitism.

    2018 04 v.31 [Abstract][OnlineView][Download 890K]

  • Gene expression changes during osteogenic and adipogenic differentiations of rat bone marrow mesenchymal stem cells

    YIN Xiao-li;LU Yan-qin;LIU Bao-yan;HAN Jin-xiang;School of Medicine and Life Sciences,University of Jinan-Shandong,Academy of Medical Sciences;

    Objective To investigate the change of expression of relevant genes during osteogenic and adipogenic differentiations of rat bone marrow mesenchymal stem cells. Methods BMSCs from rats were isolated and cultured by adherent culture,observed for morphological change and determined for growth by MTT assay,based on which the growth curve was plotted. The osteogenic and adipogenic differentiations of BMSCs of passage 4 were induced with the corresponding inducers,and the alkaline phosphatase(ALP)activity,osteogenic and adipogenic differentiation abilities of the cells were determined by ALP kit,alizarin red staining and oil red staining respectively. The mRNA expressions of Runx2,osteocalcin(OCN),ALP,peroxidase proliferator activated receptor gamma(PPARγ) and fatty acid binding protein 4(FABP4) after induction for 0,7,14 and 21 d were determined by RT-QPCR. Results BMSCs were successfully isolated and cultured. Passaged cells were proliferated rapidly,most of which were in a long spindle form.The cell growth curve was in an S shape. After induction with osteogenic and adipogenic inducers,the results of ATP activity test,alizarin red staining and oil red staining of BMSCs of passage 4 were positive. The expression levels of mRNA of osteogenic genes(Runx2,OCN and ALP)and adipogenic genes(PPARγ and FABP4)after induction for 7,14 and 21 d were significantly higher than those before induction(0 d)(P < 0. 05). During osteogenic differentiation,the expression levels of Runx2 and ALP reached the maximums after induction for 7 d then showed decreasing trends,while that of OCN showed a steady increasing trend. However,during adipogenic differentiation,the expression level of PPARγreached the maximum after induction for 7 d,while that of FABP4 was higher throughout. Conclusion BMSCs are easy to be isolated,cultured and amplified in vitro,with potential of multi-differentiation after induction. The expression levels of genes related to osteogenic and adipogenic differentiations changed with the increasing time for induction,indicating the regulatory roles of the genes. It provides an experimental basis for study on the mechanism of BMSCs in bone,cell and gene engineering.

    2018 04 v.31 [Abstract][OnlineView][Download 465K]

  • Effects of lipopolysaccharide on proliferation rate of bovine mammary fibroblasts as well as mRNA expression levels of intracellular Toll-like receptor and its signal pathway-related genes

    ZHANG Xu;WU Rui;WANG Jian-fa;HE Xian-jing;YANG Bin;XU Dan-dan;WANG Hai;SHAN Xu-fei;WANG Ying-qi;YUE Yang;College of Animal Science and Veterinary Medicine, Heilongjiang Bayi Agricultural University;

    Objective To investigate the effects of lipopolysaccharide(LPS) on proliferation rate of bovine mammary fibroblasts(BMFBs) as well as mRNA expression levels of intracellular Toll-like receptor and its signal pathway-related genes. Methods BMFBs were subjected to primary culture and stimulated with LPS at a final concentration of 10 μg/mL for various hours. The proliferation rate of BMFBs was determined by CCK-8 kit, while the expression levels of Toll-like receptor and its signal pathway-related molecular mRNAs by real-time fluorescent quantitative PCR. Results The proliferation rates of BMFBs showed no significant difference after stimulation with LPS for 0, 1 and 3 h(P > 0. 05), while was significantly higher in test group than in control group more than 6 h after stimulation(P < 0. 05). As compared with those in control group, the expression levels of TLR2, TLR4, signal pathway-activated nuclear factor kappa B(NF-κB),tumor necrosis factor(TNF) receptor associated factor 6(TRAF6), interleukin-1β(IL-1β), IL-6, IL-8, TNFα and interleukin-1 receptor associated kinase 1(IRAK1) mRNAs 12 h after stimulation increased significantly(P < 0. 05),while that of myeloid differentiation factor 88(MyD88) mRNA showed no significant change(P > 0. 05). Conclusion BMFB might recognize LPS by TLR2 and TLR4 and cascade with NF-κB signaling pathway to induce the release of inflammatory cytokines and chemokines, and participate in dairy mammary innate immune responses.

    2018 04 v.31 [Abstract][OnlineView][Download 394K]

  • Effect of extracellular regulated protein kinases 1/2 on infection of myocardial cell line H9C2 with eosinophilic coxsackie virus B3

    DOU Zhong-xia;ZHU Hua;WANG Ju;Inner Mongolia People's Hospital;

    Objective To investigate the effect of extracellular regulated protein kinases 1/2(ERK1/2)on infection of myocardial cell line H9C2 with eosinophilic coxsackie virus B3(CVB3). Methods The working concentration of CVB3 was optimized by determination of TCID_(50) of CVB3 to H9C2 cells. H9C2 cells were treated with CVB3 at the optimal concentration for 0, 20, 40 and 60 min respectively, and determined for phosphorylation of ERK1/2. H9C2 cells were treated with Ceramide, an activator of ERK1/2 phosphorylation, then added with CVB3 and determined for TCID_(50).Results The TCID_(50) of CVB3 to H9C2 cells was 10~(-7. 1). The expression levels of phosphorylated ERK1/2 in H9C2 cells after treatment with CVB3 at a concentration of 10~(-7). 1 for 40 and 60 min were significantly lower than that before treatment(P < 0. 01). However, after addition with Ceramide C6, the TCID_(50) of CVB3 to H9C2 cells was 10~(-6. 47). Conclusion ERK1/2 phosphorylation inhibited the infection of H9C2 with CVB3, which laid a foundation of study on the pathogenic mechanism of viral myocarditis.

    2018 04 v.31 [Abstract][OnlineView][Download 230K]

  • Establishment of national serum standard for antigen activity test of house dust mite products

    YANG Ying-chao;LI Zhe;CHEN Jian-jun;MA Xiao;XIN Xiao-fang;National Institutes for Food and Drug Control;

    Objective To establish the national serum standard for determination of antigen activity of house dust mite(HDM)products. Methods Patients allergic to HDM were confirmed by clinical symptoms and serological test,of whom the serum samples were collected and mixed,and determined for the specific IgE content against HDM by ImmunoCAP assay. The specific IgE values for HDM were determined by calibrated co-ordinately. The samples provided by the manufacturers were incubated with the national serum standard and in-house serum reference respectively, and determined for the activity of HDM by ImmunoCAP assay. Results In the national serum standard,the specific IgE contents against Dermatophagoides pteronyssinus and Dermatophagoides farina were 247. 4 and 243. 4 KUA/L respectively. Conclusion The first batch of national serum standard for determination of antigen activity of HDM products in China has been established and approved by the National Committee for Drug Standard Substances,of which the lot number is 290002-201601.

    2018 04 v.31 [Abstract][OnlineView][Download 196K]

  • Development of key methods for quality control of recombinant humanized anti-programmed death-1 monoclonal antibody

    GUO Qiu-ju;LI Xue;XU Zhi-yuan;CHEN Hong-xia;YU Yu-gen;The Antibody Center of Main Luck Pharmaceutical Co., Ltd.;

    Objective To develop the key methods for quality control of recombinant humanized anti-programmed death(PD)-1 monoclonal antibody(mAb). Methods Three batches of recombinant humanized anti-PD-1 mAb were identified by reversed phase high performance liquid chromatography(RP-HPLC) after digestion with trypsin, and determined for purity by reduced/non-reduced capillary electrophoresis-dodium dodecyl sulfonated(CE-SDS), for the contents of monomer and polymer by size exclusion high performance liquid chromatography(SEC-HPLC), for isoelectric point by capillary isoelectric focusing(c IEF), for the content of charge isomers by cation exchange high performance liquid chromatography(CEX-HPLC), and for biological activity by mixed lymphocyte reaction and competitive ELISA. Results The peptide maps of three batches of recombinant humanized anti-PD-1 mAb were consistent with that of reference, of which the purities were 97. 8%, 98. 1% and 97. 9% by reduced CE-SDS, and 95. 7%, 95. 4% and 95. 5% by nonreduced CE-SDS, respectively. However, the monomer contents of the three batches were 99. 3%, 99. 1% and 99. 4%,while the polymers were 0. 5%, 0. 6% and 0. 4%, respectively,and all the isoelectric points of the main peak shown by cIEF were 7. 4. The contents of acidic domains of the three batches were 21. 8%, 21. 4% and 21. 6%, while the main peak contents were 58. 4%, 58. 6% and 58. 7%, the contents of basic domains were 19. 8%, 20. 0% and 19. 7%, and the relative biological activities were 112%, 98% and 102%, respectively. Conclusion The developed method may be used for the routine quality control of recombinant humanized anti-PD-1 mAb.

    2018 04 v.31 [Abstract][OnlineView][Download 420K]

  • Prokaryotic expression of human cytokeratin 20 protein and preparation of monoclonal antibody

    SONG Jun-ying;YUAN Yong;ZHANG Zhong-yun;YAN Min;ZENG Hua-hui;LIU Wen-di;ZHANG Zhen-qiang;Immunological Laboratory of Traditional Chinese Medicine,Henan University of Traditional Chinese Medicine;

    Objective To express human cytokeratin 20(CK20)in prokaryotic cells and prepare its monoclonal antibody(McAb). Methods The exon gene of CK20 was amplified by using the cDNA of CK20 and inserted into prokaryotic expression vector pET28 a. The constructed recombinant plasmid p ET28 a-ck20 was transformed to E. coli BL21 for expression under induction of IPTG. The expressed recombinant CK20 was purified by affinity chromatography,with which BALB/c mice were immunized to prepare McAb by hybridoma technique. Hybridoma cells were screened by indirect ELISA,and McAb was prepared,analyzed for subtype and identified by Western blot and immunohistochemical assay. Results Restriction analysis and sequencing proved that recombinant plasmid pET28 a-ck20 was constructed correctly. The purified CK20 reached a purity of more than 90%. The prepared McAb consisted of an IgG2b heavy chain and a Kappa light chain,which showed specific binding to CK20 antigen,even at a dilution of 1:200. In immunohistochemical assay,the color of colon cancer containing CK20 after reaction with the McAb was darker than that after reaction with the imported McAb. Conclusion The McAb prepared by the expressed human CK20 may be used for determination of expression of the protein in the relevant tissues,which provides an effective tool for clinical diagnosis and research of CK20.

    2018 04 v.31 [Abstract][OnlineView][Download 443K]

  • Effectiveness of emergency vaccination with varicella vaccine at different time intervals after appearance of the first cases of varicella

    ZHANG Nan;LI Zhi;XU Na;JIANG Da-lei;LIU Jie-chen;REN Jia;TAO Hang;SUN Xiao-dong;Changchun BCHT Biotechnology Co.;

    Objective To analyze the effectiveness of emergency vaccination with varicella vaccine at different time intervals after occurrence of the first cases of varicella. Methods During the period of June 2013 to December 2015,the students from nursery,primary school,middle school,high schools and secondary vocational and technical school in Shanghai City,who met the requirements in Protocol for Emergency Varicella Vaccination in Shanghai City,were selected as the subjects,vaccinated with varicella vaccine at different time intervals(0 ~ 3 d,4 ~ 7 d,8 ~ 14 d,15 ~21 d and not less than 22 d)after occurrence of the first case of varicella,and observed for the secondary cases. Results The occurrence rate of secondary cases was higher in the subjects vaccinated at an interval of 0 ~ 3 d after the occurrence of the first case than in those at intervals of 4 ~ 7 d,8 ~ 14 d and 15 ~ 21 d(P < 0. 003),while in that at an interval of4 ~ 7 d than in those at 8 ~ 14 and 15 ~ 21 d(each P < 0. 003),and in that at 8 ~ 14 d than in those at 15 ~ 21 and not less than 22 d(each P < 0. 003). However,the attack rate of varicella was lower in the subjects with emergency inoculation at an interval of 0 ~ 3 d after occurrence of the first case than in those at 4 ~ 7,8 ~ 14,15 ~ 21 and not less than 22 d(each P < 0. 003),while in that at 4 ~ 7 d than in those at 8 ~ 14,15 ~ 21 and not less than 22 d(each P <0. 003,and in that at 15 ~ 21 d than in those at not less than 22 d(P < 0. 003). Conclusion When the varicella epidemic outbreaks,the emergency vaccination shall be carried out within 7 d after occurrence of the first case of varicella so as to effectively decrease the secondary occurrence rate and attack rate.

    2018 04 v.31 [Abstract][OnlineView][Download 153K]

  • Immune effect of booster immunization with groups A and C meningococcal polysaccharide conjugate vaccine and group A meningococcal polysaccharide vaccine by various sequential programs

    LV Hai-ying;ZHOU Hai;Zhongshan Municipal Center for Disease Control and Prevention;

    Objective To observe the immune effect of booster immunizations with groups A and C meningococcal polysaccharide conjugate vaccine(MCV-AC) and group A meningococcal polysaccharide vaccine(MPV-A) by various sequential programs. Methods A total of 118 children primarily immunized with two doses of MPV-A or MCV-AC at ages of less than 2 years were boosted with one dose of MCV-AC at ages of 3 ~ 4 years,of whom blood samples were collected before and 4 ~ 5 weeks after the booster immunization and determined for geometric mean titers(GMTs) of antibodies against groups A and C Neisseria meningitides(Nm)in sera,based on which the positive conversion rates were calculated. The study was performed by dividing the children into two groups on the basis of primary immunization with MCV-AC or MPV-A. Results Before booster immunization,the positive conversion rates of antibody against group A Nm in MCV-AC and MPV-A groups were 18. 75% and 14. 28%,while the GMTs were 2. 09 and 1. 79,respectively,which showed no significant differences(P > 0. 05). Both the positive conversion rates of antibody against group C Nm were 0 in the two groups,which showed no significant difference(P > 0. 05),while the GMTs were 1. 30 and 1. 00 respectively,which showed significant differences(P < 0. 05). After booster immunization,the positive conversion rates of antibody against group A Nm in MCV-AC and MPV-A groups were 97. 91% and 100. 00%,while the GMTs were 241. 63 and 227. 32,respectively,which showed no significant differences(P > 0. 05),and both the positive rates and GMTs in the same groups before and after booster immunization showed significant difference(P < 0. 05). However,the positive conversion rates of antibody against group C Nm in the two groups were 89. 58% and 92. 85% respectively,which showed no significant difference(P > 0. 05),while the GMTs were 106. 09 and 105. 00 respectively,which showed significant difference(P < 0. 05),and both the positive rates and GMTs before and after booster immunization showed significant difference in the same groups(P < 0. 05). Conclusion A booster immunization with MCV-AC is necessary,which induces a good immune response after primary immunization with either MPV-A or MCV-AC

    2018 04 v.31 [Abstract][OnlineView][Download 167K]

  • Application of interferon gamma release assays to patients with acid-fast bacilli positive pulmonary diseases and analysis of drug-sensitivity

    ZHAO Xi-wei;HE Li;CHEN Gang;DU Xian-zhi;Department of Respiratory Medicine, The Second Affiliated Hospital, Chongqing Medical University;

    Objective To analyze the role of interferon gamma release assays(IGRAs) in diagnosis of acid-fast bacilli positive pulmonary diseases as well as the result of drug sensitivity test so as to provide a reference for clinical treatment of the diseases. Methods The sputum, pleural effusion, tissue biopsy and alveolar lavage fluid of 325 patients with acid-fast bacilli positive pulmonary diseases in The Second Affiliated Hospital of Chongqing Medical University and the People's Hospital of Chongqing were collected from January 2015 to December 2016 and subjected to identification of mycobacterium, IGRAs and drug sensitivity test. Results Of the 325 cases acid-fast bacilli positive pulmonary diseases,308(94. 8%) were positive for Mycobacterium tuberculosis(MTB) while 17(5. 2%) for non-tuberculosis mycobacteria(NTM). The sensitivity of IGRAs to diagnosis of MTB pulmonary diseases was 85. 1%, while that to NTM pulmonary diseases was 88. 2%. The resistance rate of MTB to first-line anti-tuberculosis drugs was higher than that to second-line drugs(P < 0. 05). However, most of NTM were highly resistant to first-line anti-tuberculosis drugs. Conclusion The patients with acid-fast bacilli positive pulmonary diseases should be subjected to identification of mycobacterium and drug sensitivity test so as to treat MTB and NTM accurately.

    2018 04 v.31 [Abstract][OnlineView][Download 181K]

  • Isolation,culture and identification of Mycoplasma pneumoniae

    LI Shuang-li;ZHU Wen-yong;FENG Lei;CHAI Ling;ZHANG Jun;LI Lei;CHEN Feng;YANG Li-yuan;GUAN Xin-long;CHEN Jing-yi;LI Jie-fen;SHI Ting-ming;ZHANG Ji-hua;LI Wei-dong;LIAO Guo-yang;Institute of Medical Biology,Chinese Academy of Medical Science & Peking Union Medical College,Yunnan Key Laboratory of Vaccine Research & Development on Severe Infection Disease;

    Objective To isolate,culture and identify human Mycoplasma pneumoniae(MP) strains. Methods Mycoplasma pneumoniae strains were isolated from throat swabs and sputum samples of hospitalized children by using medium supplemented with antibiotics or filtration with filtering membrane to remove most of the fungi and bacteria,and the time for culture was optimized. The isolates were identified by PCR,genotyping as well as tests for culture characteristics,biochemical characteristics and serological characteristics. Results Most of the fungi and bacteria were removed by several pretreatment methods,such as sequential addition of antibiotics and filtration with filtering membrane. A total of 42 strains of Mycoplasma pneumoniae isolates were obtained from 978 throat swabs and sputum samples by subculture the throat swabs for 6 ~ 8 h and the sputum samples for 18 ~ 24 h after inoculation,and identified as MP from PCR characteristics,culture characteristics,biochemical characteristics and serological characteristics,of which 39 were P1 typeⅠand 3 were P1 typeⅡ. Conclusion A method for isolation and culture of MP strains from clinical specimens was developed,and 42 MP strains were successfully isolated,which were of an important application value in the research and diagnosis of MP infection and development of MP vaccine.

    2018 04 v.31 [Abstract][OnlineView][Download 379K]

  • Comparison of effectiveness of two methods for extraction during preparation of bulk of freeze-dried live attenuated hepatitis A vaccine

    HOU Li-juan;KANG Yuan-hang;ZHOU Yan-bin;WEN Xiang-hua;ZHANG Hui-ying;LI Hai-yan;LI Shu-sheng;XU Guo-liang;ZHANG Jun-min;Changchun Institute of Biological Products Co., Ltd.;

    Objective To compare the effectiveness of two methods for extraction during preparation of bulk of freezedried live attenuated hepatitis A(HA)vaccine and provide a reference for optimization of the production process. Methods The harvest of hepatitis A virus(HAV)after ultrasonication was extracted by methods A and B respectively, and the prepared bulks of vaccine were tested for sterility, mycoplasma, protein content and virus titer. Three consecutive batches of bulks and final products were prepared by the optimized method, and subjected to overall control tests according to the requirements in Chinese Pharmacopoeia(Volume Ⅲ, 2010 editioin) and the relevant internal standards. Results The protein content in bulk prepared by extraction method B was significantly lower than that by method A(P < 0. 001),while the virus titer showed no significant difference(P > 0. 05). All the quality indexes of three batches of bulks and final products prepared by extraction method B met the relevant requirements. Conclusion Both the purity of bulk and quality of vaccine were improved by extraction method B.

    2018 04 v.31 [Abstract][OnlineView][Download 164K]

  • Development of a method for amplification of Staphylococcus aureus phage

    XIE An-ping;WEI Dong;CHEN Cheng;DONG Qiao-xiang;School of Laboratorial Medicine and Life Science,Wenzhou Medical University;

    Objective To develop a method for amplification of Staphylococcus aureus phage. Methods The condition for amplification was optimized by investigating the effect of host bacterial concentration,phage titer,culture time and the existence of calcium chloride on amplification of S. aureus phage. Results The optimal concentration of S. aureus in amplification cultivation system was 1 × 10~7 bacteria/mL,while the optimal phage titer was 1 × 10~3 PFU/mL,and the optimal culture time was 6 ~ 8 h. The existence of calcium chloride showed no effect on phage amplification,and the phage titer reached a relatively high level. Conclusion A method for amplification of S. aureus phage was developed,which laid a foundation of preparation of therapeutic S. aureus phage.

    2018 04 v.31 [Abstract][OnlineView][Download 193K]

  • Evaluation on recovery of poliovirus Sabin strain after dialysis

    ZHAO Heng;CAI Qiu-long;YUAN Mei;LI Na;LI Ya-xian;YANG Jia;LIU Zheng-ling;ZHNAG Xiao-li;CAI Lu-kui;YI Li;Institute of Medical Biology,Chinese Academy of Medical Science & Peking Union Medical College;

    Objective To evaluate the recovery of polio virus Sabin strain of three types after dialysis. Methods Polio virus Sabin strain of types Ⅰ,Ⅱ and Ⅲ,at high titers,were dialyzed. The recoveries of titers of each type before and after dialysis,as well as the clearance rates of formaldehyde after dialysis,were compared. However,the polio virus at titers of 101,100 and 10-1 CCID_(50)/mL,and the samples of the same type and titer before and after dialysis were inoculated to Hep-2 cells and subcultured for three passages. The cytopathic effect(CPE)rates and virus titers before and after dialysis were compared. Results The volume and titer of samples of various types at high titers after dialysis showed no significant difference with those before dialysis(P > 0. 05),while the clearance rate of formaldehyde after dialysis was more than 99%. All the samples of various types at a titer of 101 CCID_(50)/mL caused CPE after subculture for three passages. The CPE rates caused by samples of typesⅠ,Ⅱ and Ⅲ,each at a titer of 100 CCID_(50)/mL,were 66%,33% and 33% respectively after subculture for one passage,all of which were 100% after subculture for two and three passages. However,the samples of various types at a titer of 10-1 CCID_(50)/mL caused no CPE after subculture for three passages. All the virus titers of samples after CPE were more than 6. 0 lgCCID_(50)/mL. Conclusion The recovery of three types of polio virus Sabin strain of three types were high after dialysis,with little loss,indicating that dialysis might be used for purification of the virus.

    2018 04 v.31 [Abstract][OnlineView][Download 160K]

  • Potency validation of biological activity assay for recombinant human monoclonal antibody against epidermal growth factor receptor

    JIANG Yu-jia;DENG Jing;WANG Qian;QI Hong;Simcere Pharmaceutical Company;

    Objective To perform potency validation on an established biological activity assay for human monoclonal antibody(McAb)against epidermal growth factor receptor(EGFR). Methods Four batches of McAb, one was served as control group while three as test groups, were validated. The initial concentration of McAb in control group was set as 100%, while those in test groups as 60%, 70%, 80%, 100%, 120%, 130% and 140%. The antibody activities in various groups were determined by the established biological activity assay. The results were analyzed with PLA3.0 software to adjust whether the regression, linearity, parallelism and equivalence was qualified and to calculate the measured potencies.Accuracy was expressed as the recovery rate. The linearity and range were validated by linear regression with nominal potency as the abscissa and measured potency as the ordinate. Results The regression, linearity, parallelism and equivalence in three test groups were qualified, while the mean recovery rates were in the range of 80% ~ 125%. The linear regression with nominal potencies against measured potencies showed a y intercepe of -1. 716 5, a slope of 0. 984 6 and a correlation coefficient of 0. 867 9, all of which met the relevant standards. Conclusion This method reflected the biological activity of anti-EGFR McAb at a concentration of 60% ~ 140%.

    2018 04 v.31 [Abstract][OnlineView][Download 152K]

  • Development of a method for determination of endotoxin content in extract FM902 from Angel yeast

    YAN Wei-hong;ZOU Kun;HE Hai-bo;LI Xiao;WU Ye-xu;LIU Guang-yao;CHEN Zhi-feng;WANG Jun-zhi;DENG Zhang-shuang;Hubei Key Laboratory of Natural Products Research and Development,College of Chemistry and Life Science,China Three Gorges University;

    Objective To develop a method for determination of endotoxin content in extract FM902 from Angel yeast.Methods The endotoxin content of yeast extract FM902 was determined by effectiveness test,sensitivity test,FM902 gel interference test and FM902 gel semi-quantitative test. Results The endotoxin content in extract FM902 was 5 EU/mg,which met the limit standard in Chinese Pharmacopoeia(Volumn Ⅲ,2015 edition). Conclusion A method for determination of endotoxin content in extract FM902 from Angel yeast was successfully developed,which was easy to handle and of good repeatability,and might be used for evaluation of endotoxin content in Angel yeast extract.

    2018 04 v.31 [Abstract][OnlineView][Download 190K]

  • Progress in research on role of epidermal growth factor receptor in tumorigenesis and tumor development

    ZHANG Jin-jin;TANG Hui;Institute of Basic Medical Sciences,Affiliated Hospital of Kunming University of Science and Technology;

    Epidermal growth factor receptor(EGFR),a membrane protein with tyrosine kinase activity,is expressed in human epidermal cells and stromal cells,and is highly expressed in a variety of human tumors. It is also involved in the development of a series of malignant tumors,which plays an important role in promoting the growth,proliferation,angiogenesis,invasion,metastasis and apoptosis of tumor cells,and shows a certain regulatory effect. The expression level of EGFR may be used as a prognostic indicator of various tumors. This paper reviews the progress in research on the role of EGFR in tumorigenesis and tumor development.

    2018 04 v.31 [Abstract][OnlineView][Download 219K]

  • Current status of research on application of nanobodies in tumor therapy

    WANG Zhi-ming;YANG Li-xia;NCPC New Drug Research and Development Co.,Ltd.State Key Laboratory of Antibody Drug Development;

    Given the unique molecular characteristics,nanobodies(Nbs)with low molecular mass are amenable to many applicability as diagnostic or therapeutic tools. When nanobodies were used in oncology,not much was expected from them initially because the lackof Fc-effector domain needed for cytotoxicity. However,the deepening research and clever rational design may overcome this limitation. The Nbs in research and development participate in tumor therapy in a variety of forms:direct antagonistic effects,linkage to effector moieties,incorporation into drug delivery systems,and application to radionuclide therapy and photodynamic therapy,etc. This paper reviews the drug characteristics and application if Nbs in tumor therapy as well as current status of research on Nbs drugs in clinical trial.

    2018 04 v.31 [Abstract][OnlineView][Download 237K]

  • Progress in research on enteroviruses type 68

    GAO You;XU Xing-li;LI Qi-han;Kunming Medical University;

    As a member of Picornaviridae family,species Enterovirus D,enterovirus type 68(EV-D68) is a viral pathogen isolated from a child who hospitalized and manifested pneumonia and bronchiolitis in 1962. Though few of reports about the virus have been published since its discovery,a large epidemic was observed in 2005. In 2014,a large epidemic happened in North America. This viral infection usually leads to a series of clinical symptoms including cough,wheezing and hypoxemia in children,and is considered as being associated with some cases of acute flaccid paralysis reported recently. This paper reviews the progress in research on the biological characters,epidemiology,pathogenesis and clinic features of EV-D68.

    2018 04 v.31 [Abstract][OnlineView][Download 185K]

  • Progress in research on pneumococcal surface protein A

    QIAN Jing;CAO Ling;Children's Hospital,Capital Institute of Paediatrics;

    Streptococcus pneumonia(Sp)is the main pathogen of community-acquired infection in children. At present,the long-term use of heptavalent pneumococcal conjugate vaccine and 23-valent pneumococcal polysaccharide vaccine may induce serotype substitution,which may weaken the benefits of vaccination. Looking for a new candidate vaccine is an effective way to prevent the diseases caused by Sp. This paper reviews the progress in research on pneumococcal surface protein A(PspA),the most important candidate antigen for the next generation of Sp vaccine.

    2018 04 v.31 [Abstract][OnlineView][Download 162K]

  • Progress in research on herpes simplex virus type 1 vaccine

    XU Xing-li;LI Qi-han;Institute of Medical Biology,Chinese Academy of Medical Sciences and Peking Union Medical College,Yunnan Key Laboratory of Vaccine Research and Development of Severe Infectious Disease;

    Herpes simplex virus type 1(HSV-1) infection may result in herpes labialis,genital herpes,encephalitis,and be latent in nervous system for a long time. Currently,there are no intervention measures to prevent the primary infection and control the latent and recurrent infections of HSV-1. Vaccines are still considered as the best way to control viral infection. This paper reviews the progress in research on HSV-1 vaccine.

    2018 04 v.31 [Abstract][OnlineView][Download 155K]
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@2012 Editorial Department of "Chinese Journal of Biologicals"