2021年05期目次

  • Poliovirus shedding pattern after immunization with types Ⅰ+Ⅲ oral bivalent live attenuated poliovirus vaccine in poliovirus receptor transgenic mice

    ZHAO Ting;SHI Hong-yuan;LI Jing;CHEN Shi-yi;FU Yu-ting;ZHAO Yu-ping;YANG Xiao-lei;YANG Jing-si;Institute of Medical Biology,Chinese Academy of Medical Sciences and Peking Union Medical College;

    Objective To investigate the viral shedding pattern in poliovirus receptor transgenic mice after immunization with types Ⅰ + Ⅲ oral bivalent poliovirus vaccine(bOPV). Methods The poliovirus receptor transgenic mice were inoculated with three doses of bOPV by gavage each at an interval of 28 d,of which the fecal samples were collected on days 1,3,5 and 7 after each dose and determined for viral shedding level by real-time fluorescent quantitative PCR.Poliovirus-specific IgA level in intestinal mucosa was determined by ELISA,and the relationships of viral shedding to the dose of inoculation and IgA level were analyzed. Mice with high and low viral shedding levels after inoculation with bOPV were cohabitated with clean mice respectively,and the viral shedding of clean mice was observed. Results The viral shedding level caused by the vaccine decreased gradually with the increasing dose. The mice with high viral shedding level after the first dose showed significantly lower shedding level after the second dose than those with low viral shedding level after the first dose(P < 0. 05). The mice positive for IgA after the first dose showed higher viral shedding level after the first dose,while showed lower viral shedding level after the second dose,than those negative for IgA. The probability of shedding virus in clean mice after cohabitation with high viral shedding mice was significantly higher than that with low viral shedding mice(P < 0. 05),and the viral shedding of clean mice caused by type Ⅲ poliovirus vaccine strain was more persistent than that by typeⅠ. Conclusion In the final stages of polio eradication,a key to prevent polio epidemic remains the stimulation of population mucosal immunity against poliovirus replication and the reduction of shedding.Increasing the doses of bOPV can reduce the probability of shedding after exposure to poliovirus. Viral shedding monitoring and personal hygiene should be strengthened after bOPV inoculation,especially after the first dose.

    2021 05 v.34 [Abstract][OnlineView][Download 1373K]

  • Fusion expression and immune protective effect of pertussis filamentous hemagglutinin and StxB

    LIU Zhao-lu;ZHU Li;PAN Chao;LU Ying;WANG Heng-liang;College of Food Science and Technology,Shanghai Ocean University;

    Objective To express pertussis filamentous hemagglutinin (FHA)fragment FHA_(1877-2250)fused with Shiga toxin B unit(StxB)protein and evaluate the immune protiective effect of StxB-Fha_(1877-2250)fusion protein.Methods Primers were designed to link StxB to the N-terminus of FHA_(1877-2250).The constructed recombinant plasmids pET-28a-FHA_(1877-2250)and pET-28a-StxB-Fha_(1877-2250)were transformed to E.coli BL21(DE3) and induced with IPTG.The expressed recombinant protein was purified by nickel ion affinity chromatography and re-naturalized,with which BALB/c mice were immunized s.c.for 3 times each at a interval of 2 weeks.Serum samples were collected 10 d after the last immunization and determined for antibody titer by indirect ELISA.The bactericidal efficiency was evaluated by serum bactericidal assay(SBA)in vitro.The mice were challenged i.p.with Bordetella pertussis Bp148 strain at twice median lethal dose(1.21×10~8CFU) on day 14 after the last immunization to evaluate the protective effect.Results Colony PCR and sequencing proved that recombinant plasmids pET-28a-FHA_(1877-2250)and pET-28a-StxB-Fha_(1877-2250)were constructed correctly.SDS-PAGE and Western blot showed that the relative molecular masses of expressed proteins FHA_(1877-2250)and StxB-Fha_(1877-2250)were about 43 000 and about 51 000 respectively.The serum antibody titers induced by StxB-Fha_(1877-2250)in mice was 1∶3 200.The bactericidal efficiency of serum of mice immunized with StxB-Fha_(1877-2250)was slightly higher than that with FHA_(1877-2250).Both FHA_(1877-2250)and StxB-Fha_(1877-2250)proteins showed protective effect against the challenge with Bp148 strain,of which the protection rates were 40%and 60%respectively.Conclusion Recombinant protein StxB-Fha_(1877-2250)showed better immunogenicity and protective effect than FHA_(1877-2250),which provided a new idea for the research on pertussis subunit vaccine.

    2021 05 v.34 [Abstract][OnlineView][Download 1527K]

  • Expression of Nicotiala alata defensin 1 protein in Pichia pastoris and purification of expressed product

    YANG De-yi;ZHAO Yi-chen;GONG Wei-wei;LI Kun-peng;College of Tea Science,Guizhou University,The Key Laboratory of Plant Resources Conservation and Germplasm Innovation in Mountainous Region(Ministry of Education),Guizhou Key Laboratory of Agricultural Biological Engineering;

    Objective To express Nicotiala alata defensin 1(NaD1)protein in Pichia pastoris and purify the expressed product.Methods Recombinant plasmid pPIC9 K-NaD1 was linearized with SacⅠ,electrotransformed to competent P. pastoris GS115 and induced with methanol. The single colonies with high expression level were selected for expansion culture,and the obtained recombinant NaD1 protein was purified by Ni-NTA chromatography and identified by 15% SDS-PAGE and Western blot. Results The purified recombinant protein,with a relative molecular mass of about 20 000,showed specific binding to mouse monoclonal antibody against NaD1. A specific band with relative molecular mass of about 20 000 was observed,of which the purity reached 85%. Conclusion NaD1 protein was successfully expressed in P. pastoris,which reached a high purity after purification. This study laid a foundation of further research on the mechanism of NaD1 protein in killing fungi.

    2021 05 v.34 [Abstract][OnlineView][Download 1350K]

  • Transmission and evolution of Dengue virus type Ⅰ in China

    LI Dai-ying;LU Ming;ZHU Hai-lian;HE Li-cun;GU Ran;ZHANG Juan;CHEN Jun-ying;PAN Yue;SUN Qiang-ming;Institute of Medical Biology,Chinese Academy of Medical Science & Peking Union Medical College,Yunnan Key Laboratory of Vaccine Research & Development on Severe Infectious Diseases;

    Objective To investigate the association of distribution of epidemic strain of Dengue virus(DENV)in China with the isolates in Southeast Asia,analyze the regularity of transmission,trace the source and explore the phylogenesis and molecular evolutionary characteristics of the virus. Methods Based on the number of cases published by the Public Health Science Data Center,the infection and distribution of Dengue virus type I(DENV-1)in China were analyzed,and the complete gene sequences of 405 DENV-1 strains in NCBI database since 1985 were collected,of which the sequences were compared by using the online tool MAFFT,and a phylogenetic tree was constructed by using online phyML on ATGC website. Based on the results of phylogenetic analysis, more than 40 representative strains and and the representative strains from Southeast Asia were selected and subjected to phylogenetic analysis by using MEGA X software. Afterwards,the sequences of epidemic DENV-1 strains in China were subjected to recombinant evolutionary analysis. Results From 2004 to 2017,a total of 66 623 cases of dengue fever were reported in China,among which the cases caused by DENV-1 were reported in Guangdong Province every year. Since 2012,the number of cases caused by DENV-1 increased rapidly,and the geographical scope was also expanded. The epidemic strains in China were closely associated with those in Southeast Asia. The imported cases increased continuously,and there was also possible export of DENV-1 from Guangdong Province to Southeast Asia. In Yunnan Province,epidemic DENV strains were introduced from the countries of Indochina Peninsula and then spread in Yunnan for a long time. Significant difference was observed between the imported DENV strains in Yunnan Province and Guangdong Province,which may be considered that the imported strains came from different sources. The results of recombination analysis of 24 representative strains showed that though the partial sequence of Guangdong strain MN018297 in 2017 was similar to that of Guangdong strain KP723476 in 2014,no more recombination was found. Conclusion The DENV-1 strains in China were summarized to construct a phylogenetic tree to screen and analyze the representative strains,find the main provinces from which the DENV strains were transmitted to China and the epidemic braches of DENV in China in combination with comparison of amino acid site,which provided a reference for study on the spread and evolution of DENV-1 in China.

    2021 05 v.34 [Abstract][OnlineView][Download 1629K]

  • Physicochemical properties and structural prediction of GPR173 protein associated with follicular development

    WEI Yan;WEN Chao-chao;WANG Xi;WANG Hai-long;GUO Rui;ZHAO Hong;ZHANG Dong;GONG Tao;SUN Gong-qin;YU Bao-feng;Department of Biochemistry and Molecular Biology,School of Basic Medical Science,Shanxi Medical University;

    Objective To analyze the biochemical properties and structural characteristics of GPR173 protein associated with follicular development. Methods The amino acid sequence of GPR173 was unloaded from NCBI databank,of which the physicochemical properties,signal peptide,form of secreted protein,transmembrane structure,subcellular localization,site modification,spatial structure and protein interaction network were predicated by analyzed by using a variety of bioinformatics online software. Results GPR173 was a stable basic hydrophobic protein without typical signal peptide.With seven transmembrane regions,GPR173 was neither a classical nor a non-classical secreted protein,which was most likely to act on the plasm membrane. There might be ten protein kinase phosphorylation sites and at least two O-glycosylation sites,while the secondary structure was mainly α-helix. GPR173 belonged to the 7 tm_GPCRs superfamily,and contained a 7 tmA_SREB3_GPR173 domain. Of the ten proteins with possible interaction with GPR173,SMIM20 was with the maximum possibility,while GPR20 was with the minimum one. Conclusion GPR173 protein might be involved in the G protein-coupled receptor(GPCR)signaling pathway,which provided a theoretical basis for further research on the functional mechanisms of life activities and pathogenesis of the protein.

    2021 05 v.34 [Abstract][OnlineView][Download 1624K]

  • Effect of vascular cell adhesion molecule-1 on proliferation of encephalomyocaritis virus in vitro

    LI Sheng-jun;ZHANG Hai-xia;WANG Xing-long;LI Qian;BAO Xiao-jing;LI Xiang-rong;FENG Ruo-fei;Key Laboratory of Biotechnology and Bioengineering of State Ethnic Affairs Commission,Biomedical Research Center,Northwest Minzu University;

    Objective To investigate the effect of vascular cell adhesion molecule(VCAM)-1 on proliferation of encephalomyocarditis virus(EMCV)as well as the relevant mechanism. Methods C2C12 cells were infected with EMCV at a MOI of 0. 001,in which the VCAM-1 mRNA transcription level was determined by qPCR 24 and 48 h later,while the VCAM-1 protein expression level by Western blot. C2C12 cells were transfected with recombinant plasmid pcDNA3. 1-VCAM-1 and si VCAM-1 sequence respectively,and determined for overexpression and inhibition of expression of VCAM-1 by qPCR and Western blot. The cells were infected with EMCV at a MOI of 0. 001,and determined for virus copy number and virus titer by qPCR and TCID50 respectively. The interaction of VCAM-1 and EMCV was determined by pulldown test. Results Both the mRNA transcription and protein expression levels of VCAM-1 decreased significantly with the increasing time for EMCV infection(P < 0. 001). Both the mRNA transcription and protein expression levels of VCAM-1 in C2C12 cells after transfection with recombinant plasmid pcDNA3. 1-VCAM-1 increased significantly(P < 0. 01),while both the copy number and titer of virus decreased significantly after infection with EMCV(P < 0. 001). However,both the mRNA transcription and protein expression levels of VCAM-1 in C2C12 cells after transfection with si VCAM-1 sequence decreased significantly(P < 0. 001),while both the copy number and titer of virus increased significantly after infection with EMCV(P < 0. 001). VCAM-1 showed significant interaction with virus proteins VP1 and VP2,while showed no interaction with VP3. Conclusion VCAM-1 negatively regulated the proliferation in vitro of EMCV by a possible mechanism of interactions of VCAM-1 with structural proteins VP1 and VP2 of EMCV.

    2021 05 v.34 [Abstract][OnlineView][Download 1508K]

  • Screening and analysis of SNP loci of TLR1 and CBLB genes in Chinese Holstein cows

    LIU Juan;HAN Shuo;ZOU Zi-wen;LI Xin-li;LUO Lin;SHEN Bing-lei;College of Animal Science,Heilongjiang Bayi Agricultural University;

    Objective To investigate the association between single nucleotide polymorphism (SNP) in the Toll-like receptor 1(TLR1)or Cbl proto-oncogene B(CBLB)gene and inflammatory diseases in Chinese Holstein cows.Methods The CDS region of TLR1 gene and the 18th exon of CBLB gene were amplified with DNA pool template by PCR,and the SNP loci in Chinese Holstein cows were screened.The allele frequencies of the SNP loci were estimated,and changes in m RNA secondary structure as well as physicochemical properties,hyrophobicity,hydrophilicity,N-glycosylation site,secondary and tertiary structures of protein before and after TLR1 and CBLB gene mutations were predicted by online software.Results Base substitutions at four SNP loci,i.e.C~(1424)A,A~(1475)C,G~(1496)A and G~(1550)A,were found in the CDS region of TLR1 gene,among which the C~(1424)A and G~(1496)A were missense mutations,resulting in the changes from phenylalanine(Phe)and tryptophan(Trp)to leucine(Leu)and untranslated,respectively.However,the A~(1475)C and G~(1550)A were synonymous translations.A SNP locus was found in exon 18 of CBLB gene,at which the base substitution(G~(222406)T)was a missense mutation,resulting in the change from aspartic acid (Asp)to tyrosine (Tyr).The allele frequencies of four SNP loci before and after mutation showed significant difference,while that of SNP locus showed no significant difference.Both the proteins encoded by TLR1 and CBLB genes were unstable and were hydrophobic in general.Six potential N-glycosylation sites in the protein encoded by TLR1 gene,i.e.Asn76,Asn187,Asn293,Asn306,Asn368 and Asn443,as well as four N-glycosylation sites in that encoded by CBLB gene,i.e.Asn142,Asn602,Asn744 and Asn797,were observed.Bioinformatics analysis revealed that,after the missense mutations of TLR1 and CBLB genes,the stability of secondary structure of m RNA as well as the secondary and tertiary structures of protein showed also changes.Conclusion The occurrence of five SNP loci of TLR1 and CBLB genes in Chinese Holstein cows may affect the development of inflammatory diseases in dairy cows.

    2021 05 v.34 [Abstract][OnlineView][Download 2697K]

  • Curative effect of ICG001 combined with transforming growth factor-β on carbon tetrachloride-induced liver fibrosis in mice and relevant mechanism

    REN Yi-fan;WANG Xiao-ling;NIU Yan-bang;LI Xue-wei;XIE Jun;SHI ZHI-yong;XU Jun;Shanxi Medical University;

    Objective To investigate the curative effect of inhibitor ICG001 combined with cytokine transforming growth factor-β (TGF-β) on carbon tetrachloride-induced liver fibrosis in mouse model as well as the relevant mechanism.Methods C57BL/6 J mice were randomly divided into control,model,ICG001 and ICG001 combined with TGF-β(IT)groups. The mice in model,ICG001 and IT groups were injected i. p. with 20% carbon tetrachloride,0. 1 mL for each,while those in control group with physiological saline,0. 2 mL for each,2 times per week for 6 weeks. From the 5 th week after modeling,the m ice in ICG001 and IT groups were injected i. p. with ICG001,0. 1 mL for each,while those in control and model groups with physiological saline,0. 2 mL for each,once a day for 2 weeks. Meanwhile,the mice in IT group were injected i. p. with TGF-β,0. 1 mL for each,once 2 days for 2 weeks. At the end of the 6 th week,the mice were weighed,of which the serum samples and liver tissue were collected. The alanine aminotransferase(ALT) and aspartate aminotransferase(AST) levels in sera were determined by ELISA,while the liver tissue was observed for pathological change by microscopy with HE and reticular fiber staining. The transcription levels of mRNAs of inflammatory factors(IL-6,IL-10 and TNF-α)and fibrosis indexes(α-SMA,Col-Ⅰ and FOXO1)in liver tissue were determined by QPCR,while the expression levels of α-SMA,Col-Ⅰand FOXO1 by Western blot. Results The AST and ALT contents in model group were significantly higher than those in control group(each P < 0. 01). However,both the contents were significantly lower in ICG001 group than in model group,and in IT group than in ICG001 group(each P < 0. 01).Histological examination showed pathological remission in both ICG001 and IT groups,which was more obvious in the latter than in the former. Q-PCR showed that,compared with those in model group,the transcription levels of IL-6,TNFα,α-SMA and Col-Ⅰ mRNAs in ICG001 group decreased significantly(each P < 0. 01),while that of IL-10 mRNA increased significantly(P < 0. 05),and that of FOXO1 mRNA showed no significant difference(P > 0. 05). However,compared with those in ICG001 group,the transcription levels of IL-6,TNFα,α-SMA and Col-Ⅰ mRNAs in IT group decreased significantly(each P < 0. 01),while those of IL-10 and FOXO1 mRNAs increased significantly(each P <0. 05). Western blot showed that,compared with those in model group,the expression levels of α-SMA and Col-Ⅰproteins in ICG001 group decreased significantly(each P < 0. 01),while that of FOXO1 protein showed no significant difference(P > 0. 05). However,compared with those in ICG001 group,the expression levels of α-SMA and Col-Ⅰproteins in IT group decreased significantly(each P < 0. 05),and that of FOXO1 protein increased significantly(P <0. 05). Conclusion Compared with ICG001 alone,ICG001 combined with TGF-β improved the carbon tetrachlorideinduced liver fibrosis significantly,which might be related to the change of FOXO1 caused by the action of ICG001 on TGF-β pathway.

    2021 05 v.34 [Abstract][OnlineView][Download 1904K]

  • Protective mechanism of Mongolian medicine cinnamon on apoptosis of vascular endothelial cells induced by lipopolysaccharide

    SONG Li-hua;REN Xiang-yu;YANG Hang;YIN Zhao-li;SUN Xiao-lin;LI Lian;FU Quan;BAO Li-li;Department of Basic Medicine,Inner Mongolia Medical University;

    Objective To investigate the protective effect of Mongolian medicine cinnamon on apoptosis of vascular endothelial cells induced by lipopolysaccharide(LPS) as well as the relevant mechanism. Methods Human umbilical vein endothelial cells(HUVECs) were used as the research object. CCK-8 cell viability assay was used to screen the concentration of LPS for induction of apoptosis in vitro and the concentration of cinnamon administered. The HUVECs were divided into control(serum-free medium),LPS(1 μg/mL)and LPS(1 μg/mL)+ cinnamon(2 mg/mL)groups,of which the survival rate was determined by CCK-8 method,while the TNF-α content in culture supernatant by ELISA,the transcription levels of PARP(116 kD)and UCP2 mRNAs by real-time fluorescence quantitative PCR,and the protein expression levels of PARP(89 kD) and p-mTOR by Western blot. Results Compared with those in control group,the cell survival rate in LPS group decreased significantly(P < 0. 05),while the secretion of TNF-α increased significantly(P < 0. 01),the mRNA transcription levels of PARP(116 kD)and UCP2 decreased significantly(P < 0. 05),and the expression levels of PARP(89 kD)and p-mTOR increased significantly(P < 0. 01). However,compared with those in LPS group,the cell survival rate in LPS + cinnamon group increased significantly(P < 0. 05),while the secretion of TNF-αdecreased significantly(P < 0. 01),the mRNA transcription levels of PARP(116 kD) and UCP2 increased significantly(P < 0. 05),and the expression levels of PARP(89 kD)and p-mTOR decreased significantly(P < 0. 01). Conclusion Mongolian medicine cinnamon has a certain protective effect on apoptosis of HUVECs induced by LPS,which may play a role through mitochondrial pathway.

    2021 05 v.34 [Abstract][OnlineView][Download 1790K]

  • Specificity of 2019-nCoV nucleic acid detection kit(fluorescence PCR)

    DONG Jiang-kai;DU Jin-feng;CHENG Tian-ling;HUANG Ying-yan;XIA Xiao-kai;ZOU Yong;ZHANG Yun-tao;YANG Xiao-ming;Shanghai Geneo Dx Biotech Co.,LTD.;

    Objective To evaluate the specificity of three consecutive batches of 2019-nCoV nucleic acid detection kit(fluorescence PCR)manufactured by Shanghai GeneoDx Biotech Co.,LTD. Methods A total of 55 common respiratory pathogens,including endemic human coronaviruses(HKU1,OC43,NL63 and 229 E),severe acute respiratory syndrome conronavirus(SARS-CoV),Middle East respiratory syndrome coronavirus(MERS),seasonal influenza virus,rhinovirus,adenovirus,Klebsiella pneumonia and Mycobacterium tuberculosis,were grouped and used for verification of cross reactivity of the detection kit. According to the requirements in the Key Points of Technical Review for Registration of2019 New Coronavirus Nucleic Acid Detection Reagents issued by Center for Medical Device Evaluation,National Medical Products Administration(NMPA),human mucoprotein,human blood,phenylephrine,oxymetazoline,sodium chloride(including those as preservatives)and other 29 interfering substances were selected to verify the anti-interference substances of this kit. Results In the verification for cross reactivity,the test results of mixed positive samples by the three consecutive batches of kit were positive,while those of negative samples were negative,indicating a coincidence rate of accuracy of 100%. The cross-reactive substances showed no effect on the test result by the kit. All the test results of positive and borderline positive samples were positive,while those of negative samples were negative,indicating a coincidence rate of accuracy of 100%. All the 29 kinds of endogenous/exogenous interfering substances showed no influence on the test results by this kit. Conclusion The new coronavirus 2019-nCoV nucleic acid detection kit(fluorescence PCR method)manufactured by Shanghai GeneoDx Biotech Co.,LTD. showed no cross-reactivity with 55 common respiratory pathogens,while showed anti-interference properties against endogenous and exogenous interfering substances such as host tissue,common respiratory pathogen and common drugs for respiratory diseases in clinic. The specificity test result of the kit met the requirements for registration of 2019 new coronavirus nucleic acid detection reagents. The test results of three consecutive batches of kits were highly stable.

    2021 05 v.34 [Abstract][OnlineView][Download 1486K]

  • Prokaryotic expression of early secretory protein MPT64 from Mycobacterium tuberculosis and its application in serological diagnosis of tuberculosis

    QU Rong;WU Kang;WU Juan;FAN Xiao-yong;LYU Jian-xin;School of Laboratory Medicine and Life Science,Wenzhou Medical University;

    Objective To express the early secretory protein MPT64 from Mycobacterium tuberculosis(Mtb) in prokaryotic cells,purify the recombinant protein and evaluate its significance in serological diagnosis of tuberculosis(TB).Methods The mpt64 gene was amplified from the genome of Mtb standard strain H37 Rv by PCR and cloned into prokaryotic expression vector pET28 a(+). The constructed recombinant plasmid pET28 a-mpt64 was transformed to E. coli BL21(DE3) for expression under induction of IPTG. The expressed recombinant protein was purified by Ni2+-NTA chromatography,with which New Zealand white rabbits were immunized s. c. at a dosage of 600 μg in several sites,every other week for 4 times. Two weeks after the last immunization,blood samples were collected from ear vein,from which sera were separated,determined for antibody titer by ELISA,and identified for antigen specificity by Western blot. Serum samples were collected from 44 patients with TB and 36 healthy volunteers and determined for IgG and IgA levels against MPT64 by indirect ELISA,using Gro ES and Ag85 A as control antigens. Results Both restriction analysis and sequencing proved that recombinant plasmid pET28 a-mpt64 was constructed correctly. The recombinant MPT64 protein was mainly expressed in the form of inclusion body and partly in soluble form,of which the purity reached 90% and the protein concentration was about 2. 0 mg/m L after purification by Ni~(2+)-NTA chromatography. The rabbit polyclonal antiserum against MPT64 reached a titer of 1 ∶ 2 048 000,which showed reaction with the purified MPT64 protein and formed a specific protein band with relative molecular mass of about 24 000. Both the IgG and IgA levels against MPT64 in patients with TB were significantly higher than those in healthy volunteers as control(P < 0. 05),while those against Gro ES and Ag85 A showed no significant difference(P > 0. 05). Conclusion Recombinant MPT64 showed good efficiency in serological diagnosis of TB,which might be used as a candidate antigen for early serological diagnosis of the disease.

    2021 05 v.34 [Abstract][OnlineView][Download 1765K]

  • Investigation on carrying status of Neisseria meningitidis in healthy population in Baoshan City,Yunnan Province,China

    CHEN Chen;REN Yuan;GUO Ni;ZHENG Yue;LI Yu-zhong;TANG Cong;TAN Yin-zhen;LI Jiao-chun;WANG Xue-fu;CUN Wei;BI Yan-wei;Institute of Medical Biology,Chinese Academy of Medical Sciences;

    Objective To investigate the carrying rate of Neisseria meningitidis(Nm) in healthy population gathered in schools in Baoshan City,Yunnan Province,China,and to provide a reference for analysis of the development trend of meningococcal meningitis in the city. Methods A total of 1 076 throat swab specimens were collected from the healthy people at ages of 18～22 years in Baoshan City,from which Nm was routinely separated,cultured and identified by PCR. Results Seventeen strains of Nm were isolated from 1 076 throat swab specimens,indicating a carrying rate of 1. 58%,among which 14 positive strains belonged to group B,2 to group C,and 1 to group X. Conclusion There are asymptomatic carriers of meningococcal meningitis in the campus of Baoshan City,major of which were of group B,followed by group C,and a Nm of group X was isolated.

    2021 05 v.34 [Abstract][OnlineView][Download 1688K]

  • Meta-analysis of risk factors of herpes simplex virus type 1 infection

    FENG Xiao;XU Xing-li;LI Qi-han;Institute of Medical Biology,Chinese Academy of Medical Sciences and Peking Union Medical College;

    Objective To explore the global risk factors of herpes simplex virus type 1(HSV-1)infection so as to provide a reference for the epidemiological research and defense control of HSV-1 in China. Methods The research results of new HSV-1 infections and risk factors published worldside from 1980 to 2018 were summarized and analyzed comprehensively by meta-analysis. Heterogeneity test,combined effect quantity statistics analysis,bias test and sensitivity analysis were performed by using Review Manager5.3 software. Results The risk factors of HSV-1 infection included female(OR = 1. 33,95% CI:1. 20 ～ 1. 47,P < 0. 001),age of more than 40 years(OR = 2. 99,95% CI:1. 84 ～ 4. 86,P < 0. 001),low age of first sexual intercourse(OR = 1. 22,95% CI:1. 09 ～ 1. 36,P < 0. 001),low educational level(OR = 1. 68,95% CI:1. 23 ～ 2. 29,P = 0. 001),low income(OR = 2. 25,95% CI:1. 38 ～ 3. 66,P = 0. 001).Previous infection with HSV-2(OR = 0. 96,95% CI:0. 84 ～ 1. 11,P > 0. 5) was unrelated to the new infection.Conclusion Education on sexual knowledge and viral infection including HSV-1 should be strengthened in women,adolescents and population at the bottom of society. According to the risk degree of HSV-1 infection at different ages,a vaccination plan for clinical trials of candidate vaccines may be formulated.

    2021 05 v.34 [Abstract][OnlineView][Download 2991K]

  • Establishment of national control for protein of primary hamster kidney cells used for rabies vaccine for human use

    SHI Lei-tai;LI Jia;ZHANG Yue-lan;WANG Hui;LEI Ji-jun;GUO Zhong-ping;LI Yu-hua;National Institutes for Food and Drug Control;

    Objective To establish the national control for determination of host cell protein residues in rabies vaccine(hamster kidney cells)for human use(ELISA method). Methods The primary hamster kidney cell protein control was collaboratively calibrated and assigned by ELISA using the standard for primary hamster kidney cell protein of which the measurement was 4. 0 μg/mL,and evaluated for uniformity and stability. Results The primary hamster kidney cell protein control was assigned as 34. 7 ng/mL,of which the 95% reference range was(34. 7 ± 10. 1) ng/mL after statistical processing of the collaboratively calibrated data. The uniformity and stability of the control also met the relevant requirements. Conclusion National primary hamster kidney cell protein control with a lot number of 250025-201901 was established,which ensured the accuracy and reliability of detection system.

    2021 05 v.34 [Abstract][OnlineView][Download 1455K]

  • Development of enzyme-linked lectin assay for activity and subtyping of influenza virus neuraminidase

    CAO Hai-dan;LIU Xu-guang;WANG Ping;ZHAO Dan-hong;SUN Yu;SUN Zhi-wei;YANG Hong-yu;LI Shuai;ZHANG Xue-mei;WU Ye-hong;Changchun Institute of Biological Products Co.,Ltd.;

    Objective To develop an enzyme-linked lectin assay(ELLA) for activity and subtyping of neuraminidase(NA)of influenza virus. Methods ELLA was developed using fetal globulin-coated microtiter plate as NA substrate and horseradish peroxidase-labeled peanut agglutinin(HRP-PNA)as enzyme conjugate,and verified for durability,specificity and suitability. Results The bulk of influenza virus split vaccine showed NA activity which was in agreement in various subtypes and batches. The NI_(50) of subtype H1N1 against NA antisera N1,N2 and B were 1∶2 560,1∶80 and 1∶10,while those of subtype H3N2 were 1∶160,1∶2 560 and 1∶10,and those of subtype B were 1∶40,1∶40 and 1∶320,respectively. The NA antisera significantly inhibited the NA activity of influenza virus of the corresponding subtype.The incubation time of HRP-PNA and the additives during vaccine production showed no significant influence on the test result. The subtyping results of various vaccine virus strains of the same subtype were basically in agreement. Conclusion An ELLA was successfully developed,which might be used for determination of NA activity and subtyping of influenza virus. It provided a novel method for quality evaluation of domestic influenza virus split vaccine.

    2021 05 v.34 [Abstract][OnlineView][Download 1713K]

  • Development and verification of a method for determination of residual Triton X-100 content in quadrivalent influenza virus split vaccine by high performance liquid chromatography and comparison with turbidimetric method

    FAN Xue;WU Zheng;WANG Yi-ping;LI Xuan;ZHOU Li-bao;Research and Development Department,Liaoning Cheng Da Biotechnology Co.Ltd.;

    Objective To develop and verify a high performance liquid chromatography(HPLC)method for determination of residual Triton X-100 content in quadrivalent influenza split vaccine, and compare with turbidimetric method.Methods The condition for HPLC was determined based on the optimization of ratio of water to methanol in mobile phase(30 ∶ 70,20 ∶ 80 and 10 ∶ 90),flow rate(1. 0,1. 2 and 1. 4 mL/min)and sample loading(5,10 and 20 μL). The developed HPLC method and turbidimetric method were verified separately and compared. Results The condition of HPLC was as follows:BDS HYPERSIL C18 column(250 mm × 4. 6 mm,5 μm) was adopted,serving the mixture of water and methanol(20 ∶ 80)as mobile plase at a flow rate of 1. 2 mL/min. The detection wavelength,sample loading and column temperature were 230 nm,10 μL and 30 ℃ respectively. The Triton X-100 and the impurities were separately well by HPLC,of which the linear range of curve was 2 ～ 100 μg/mL(r = 0. 999 9). The limits of detection and quantitation of the developed method were 1 and 2 μg/mL respectively,indicating high accuracy and precision.However,the linear range of turbidimetric method for determination of standard was 10 ～ 100 μg/mL(r = 0. 996 5),while the limit of quantitation was 10 μg/mL. Conclusion Compared with turbidimetric method,HPLC is more suitable for the determination of residual Triton X-100 content in quadrivalent influenza virus split vaccine.

    2021 05 v.34 [Abstract][OnlineView][Download 1605K]

  • Development and verification of a modified BCA method for determination of plasminogen content in human plasma

    HU Yong;JI De-ming;ZHAN Qian;CHEN Ke-jin;PENG Yan;ZHOU Yan-xiang;YUE Sheng-lan;LI Juan;ZHOU Zhi-jun;Sinopharm Wuhan Plasma-derived Biotherapies Co.,Ltd.;

    Objective To develop,verify and preliminarily apply a modified PierceTMBCA protein quantitative analysis kit for determination of plasminogen(Pg) content in human plasma. Methods The Pg content in human plasma was determined by the tube assay combined with microplate assay in PierceTMBCA kit. The method was validated for linear range by comparison of fitting methods of standard curves. The internal control standard was determined by the cross calibration of national standard for bovine serum albumin(BSA)and the standard BSA in PierceTMBCA kit. The feasibility of BSA as a standard in determination of Pg content was analyzed by parallelism test. The developed method was validated for accuracy,precision and selectivity,by which the Pg content during purification was determined,and the result was compared with those by Kjeldahl method and biuret method. The Pg samples taken from the whole process were determined for relative purity and specific activity by the developed BCA method. Results Quadratic function fitting showed that the linear range of the modified BCA method was 62. 5 ～ 2 000 μg/mL. The standard bovine serum albumin(BSA)included in the kit was calibrated as 2. 090 μg/mL with national standard for BSA,which might be used as an internal control st andard. When the concentration of component containing Pg was 250 ～ 2 000 μg/mL,the curve of internal control BSA showed good parallelism to that of Pg. The recovery rates of international control standard at concen-trations of 1 500,750,150 and 62. 5 as well as national at a concentration of 2 000 μg/m L were 88. 61% ～ 103. 78%,and the both the CVs in intra-and inter-assays were less than 10%. The 150 mmol/L sodium octanoate,25 g/L sucrose,5 g/L glycine and 3% arginine hydrochloride in the process buffer showed no effect on,while the 1% S/D diluted at least 40 folds showed no interference to,the determination result. The determination results of bulk and final bulk of by the modified BCA method were consistent with those by biuret method and Kjeldahl method. The specific activity of samples taken from the whole process increased from 0. 02 to 8. 42 U/mg,while the relative purity from 0. 19% to81. 94%. Conclusion The modified method for BCA protein quantitative analysis was successfully developed,which showed wide linear range and good parallelism,accuracy and precision,which might be used for the quality monitoring of intermediates as well as determination of protein content in bulk and final bulk.

    2021 05 v.34 [Abstract][OnlineView][Download 1860K]

  • Latest progress and prospect of development of COVID-19 vaccine

    LI Xiao-rui;LI Xing-hang;YAN Han-chi;School of Life Sciences,Tianjin University;

    The emerging Coronavirus Disease 2019(COVID-19)pandemic poses a massive crisis to global public health.World Health Organization(WHO)declared the global pandemic of COVID-19 on March 11,2020. The progress of 2019-nCoV vaccines cover nearly all forms of current vaccine research,including inactivated vaccine,recombinant protein vaccine,viral vector-based vaccine,nucleic-acid vaccine and live attenuated vaccine,as well as the vaccine design based on novel concepts such as reverse vaccinology and vaccinomics. This article reviews the COVID-19 vaccines in development and clinical trials as well as the challenge in vaccine development.

    2021 05 v.34 [Abstract][OnlineView][Download 1561K]

  • Application of convalescent plasma from patients with COVID-19 in treatment of Coronavirus Disease 2019

    WANG Yue;LI Ce-sheng;YANG Xiao-ming;Sinopharm Wuhan Plasma-derived Biotherapies Co.,Ltd;

    The pandemic of Coronavirus Disease 2019(COVID-19)in 2020 has posed a great challenge to global public health resources. Since there are no specific antiviral drugs at present,convalescent plasma(CP)from patients who have recovered from COVID-19 is one of the specific biologic therapies being considered to treat severe acute respiratory syndrome coronavirus 2(SARS-CoV-2) infection. Preliminary studies have shown that the CP containing high titer neutralizing antibody against SARS-CoV-2 is safe and promising in blocking viral replication and improving patients′clinical symptoms. In this article,we briefly summarize the application of CP in treatment of COVID-19,and explores possible action mechanism,relevant clinical research and possible influencing factors of clinical effect,which may be helpful to the rational application of CP in treatment of COVID-19.

    2021 05 v.34 [Abstract][OnlineView][Download 1567K]

  • Progress in research on treatment of ischemic stroke with mesenchymal stem cells

    ZHOU Li;BAI Xue;YANG Qin;Department of Neurology,The First Affiliated Hospital of Chongqing Medical University;

    Ischemic stroke is a common and frequently-occurring cerebrovascular disease that seriously affects human health. However,most patients benefit less from the current clinical treatment. Stem cell transplantation is an emerging treatment and has been studied in various central nervous system diseases. At present,among the transplanted stem cells,mesenchymal stem cells(MSCs)are most widely used. More and more studies in animal models show that transplantation with MSCs can alleviate neurological deficits,which brings hope for the teratment of ischemic stroke. This article reviews the biological characteristics of MSCs and discusses the mechanism and progression of mesenchymal stem cell transplantation,in order to provide new therapeutic directions for ischemic stroke.

    2021 05 v.34 [Abstract][OnlineView][Download 1618K]

  • Progress in research on biomarkers in systemic lupus erythematosus

    GUO Sheng-hua;WANG Jian;The Fourth Laboratory,National Vaccine & Serum Institute;

    Systemic lupus erythematosus(SLE) is a systemic autoimmune disease,which is characterized by abnormal production of antibodies,immune complex deposition and multiple organ damage,but its pathogenesis is not clear at present. Recent studies suggest that SLE is the result of the interaction of genetic susceptibility,hormones,environment and immune system. With the development of detection technology,SLE related genes,cytokines and signaling pathways have been found gradually,which provides a new way for clinical diagnosis and treatment. This review briefly describes the biomarkers found at present from three aspects:genetic susceptibility genes,clinical diagnosis and disease affected organs,and explores the difficulties and feasible solutions from experimental biomarkers to clinical application.

    2021 05 v.34 [Abstract][OnlineView][Download 1594K]

  • Progress in research on cytotoxicity of silver nanomaterials and induced autophagy

    CHANG Yu-ling;NIU Ya-qian;LIU Fang;CHEN Che;Department of Clinical Medicine,Gansu University of Traditional Chinese Medicine;

    Nanomaterials refer to materials that have at least one dimension in the nanometer size(0. 1 ～ 100 nm) or are composed of basic units in a three-dimensional space. Silver nanoparticles(AgNPs) are one of the important nanomaterials,which play an important role in nanoscience and nanotechnology,especially in the field of nanomedicine,including potential applications in the diagnosis and treatment of cancers. AgNPs can produce cytotoxicity by targeting tumor cells specifically,therefore has a good application potential as anti-tumor reagent. However,AgNPs can activate tumor cell autophagy,cause autophagosome-lysosome fusion and result in the defect in autophagy flux,which may form a protection and impact the cytotoxicity to tumors. Therefore,this article reviews the mechanism of cytotoxicity of AgNPs and the anti-tumor mechanism of autophagy induced by AgNPs.

    2021 05 v.34 [Abstract][OnlineView][Download 1566K]


@2012 Editorial Department of "Chinese Journal of Biologicals"