2024年02期目次

  • Structural stability of Hib capsular polysaccharide in vaccine manufacturing

    ZHANG Tengteng;QIN Chunjun ;YIN Shanshan;LIU Cui;LIU Jiankai;YIN Jian ;TONG Wei;Beijing Minhai Biotechnology Co.,Ltd.;

    Objective To evaluate the stability of polyribosylribitol phosphate(PRP),the basic structure of capsular polysaccharide of Haemophilus influenzae type b(Hib),in the preparation of Hib conjugate vaccine.Methods The structures of the prepared Hib polysaccharides,polysaccharide derivatives and protein-conjugated polysaccharides were analyzed by nuclear magnetic resonance spectroscopy(NMR).Results The detection results of the prepared Hib polysaccharides,polysaccharide derivatives and protein-conjugated polysaccharides all met the requirements of relevant standards of Chinese Pharmacopoeia(VolumeⅢ,2020 edition),and the NMR spectra showed no significant change.Conclusion The basic structure PRP of the main carbohydrate antigen of Hib conjugate vaccine had no change during the vaccine manufacturing.

    2024 02 v.37 [Abstract][OnlineView][Download 1202K]

  • Expression and identification of recombinant adenovirus type 5-Norovirus GⅡ.4-VP1 virus-like particles

    WANG Mengjun;SUN Nan;PEI Jie;LIU Ruilun;ZOU Wenqi;XIONG Yu;WANG Wenhui;YU Daiguan;SHEN Shuo;Viral Vaccine Research Laboratory,Wuhan Institute of Biological Products Co.,Ltd.;

    Objective To express and identify recombinant adenovirus type 5-Norovirus(NoV)GⅡ.4-VP1 virus-like particles(VLPs)in 293T cells. Methods The recombinant adenovirus plasmids pAd5-eGFP and pAd5-NoV-GⅡ.4-VP1 were transfected into 293T cells respectively,and the recombinant adenovirus rAd5-eGFP and rAd5-NoV-GⅡ.4-VP1 were rescued. The rAd5-eGFP was subcultured in 293T cells to verify the function of the vector. The rAd5-NoV-GⅡ.4-VP1 was subcultured in293T cells,expressed and purified,and then NoV-GⅡ.4-VLP was formed by self-assembly,which was detected by Western blot,ELISA and observed by transmission electron microscope. Results The green fluorescence of the recombinant adenovirus rAd5-eGFP of various generations was observed under microscope,and the brightness increased with the increase of generations. NoV-GⅡ.4-VP1 protein was expressed in the harvested solution of recombinant adenovirus rAd5-NoV-GⅡ.4-VP1 of various generations,with a relative molecular mass of about 58 900. NoV-GⅡ.4-VLP showed specific binding to the rabbit anti-NoV-GⅡ.4-VP1 serum;it had similar conformation to natural NoV virus particles and can effectively identify NoV receptors in volunteer saliva samples;microscopic observation showed that the morphology was complete and spherical,with a diameter of 43. 5 — 58. 3 nm,while there were a few protrusions on the surface of the particles,which might be the P domain exposedon the particle surface during self-assembly of VP1. Conclusion The expressed recombinant adenovirus NoV-GⅡ. 4-VLP has complete VLP structure and good specificity,and is expected to be used in the related research of NoV adenovirus vector vaccine.

    2024 02 v.37 [Abstract][OnlineView][Download 904K]

  • Effect of microparticles derived from bone marrow mesenchymal stem cells on cardiomyocyte hypertrophy and its mechanism

    CAO Xiqian;LI Junhu;CUI Xinxin ;GUO Chenmeng;PANG Min;LIU Jiancan;BAI Xiaojie;FENG Qilong;Department of Physiology,Basic Medical College of Shanxi Medical University,Key Laboratory of Cell Physiology,Ministry of Education,Key Laboratory of Cell Physiology,Shanxi Province;

    Objective To investigate the effect of microparticles(MPs)derived from bone marrow mesenchymal stem cells(BMSCs) on myocardial hypertrophy and its mechanism.Methods The osteogenic differentiation and adipogenic differentiation of mesenchymal stem cells(MSCs) were induced. After isolation and purification,the morphological characteristics were observed by transmission electron microscope,and the MPs surface antigen was identified by flow cytometry. Myocardial hypertrophy model was induced by using isoprenaline(ISO)in rats,which were measured for the cardiac structure and function by echocardiography,and then detected for various indexes of the heart and isolated left ventricle. Single ventricular myocytes of rats were acutely isolated and divided into control group(Control group),cardiomyocyte hypertrophy group(ISO group),MPs group(MPs group),and MPs supernatant group(Supernatant group). The mRNA expressions of atrial natriuretic peptide(ANP)and B-type natriuretic peptide(BNP)were detected by qRTPCR. The expression levels of calmodulin-dependent protein kinaseⅡ（CaMKⅡ）and phosphorylated calmodulin-dependent protein kinaseⅡ（p-CaMKⅡ）were detected by ELISA. The L-type calcium current(LCa-L)in single ventricular myocyte of various groups was recorded by whole-cell patch clamp.Results The bone nodules of MSCs osteogenic differentiation turned red after alizarin red staining,and lipid droplets of adipogenic differentiation turned red after oil red O staining;Under transmission electron microscope,MPs membrane had a complete structure,a clear outline and a diameter of about200 nm;The positive rates of CD29 and CD90 on the surface of MPs were(98. 24 ± 0. 82)% and(97. 69 ± 1. 83)%,respectively. Compared with Control group,the left ventricular end diastolic dimension(LVEDD)reduced signifi-cantly(t =5. 065,P < 0. 05),while the interventricular septum end-diastolic dimension(IVSd),left ventricular posterior wall dimension(LVPWd),heart weight to body weight ratio(HW/BW),and heart weight to tibial length ratio(HW/Tibia)significantly increased in ISO group(t = 4. 013,2. 368,4. 392,5. 043 and 6. 120,respectively,each P < 0. 05),indicating that the hypertrophic model was successfully established. The expression levels of ANP and BNP mRNA in hypertrophic cardiomyocytes of rats in ISO group were significantly higher than those in Control group(t = 25. 120 and18. 261,respectively,each P < 0. 01);While the expression levels of ANP and BNP mRNA in MPs group significantly reduced after incubation with 48 μg/mL MPs for 48 h compared with ISO group(t = 12. 110 and 3. 526,respectively,each P < 0. 05);The expression levels of CaMK Ⅱand p-CaMKⅡ in ISO group were significantly higher than those in Control group(t = 3. 278 and 4. 181,respectively,each P < 0. 05),while the expression of p-CaMK Ⅱ in MPs group decreased significantly(t = 5. 420,P < 0. 05);The calcium current density in ISO group was significantly higher than that in Control group(t = 15. 261,P < 0. 01),while that in MPs group was significantly lower than that in ISO group(t =6. 216,P < 0. 05).Conclusion MSC-MPs can significantly inhibit ISO-induced cardiomyocyte hypertrophy in rats,which is related to its down-regulation of cardiomyocyte CaMKⅡ and inhibition of L-type calcium channel.

    2024 02 v.37 [Abstract][OnlineView][Download 1026K]

  • Construction of human interleukin-26 lentivirus expression plasmid and gene knockout plasmid and their activity verification in HEK293T cells

    ZHOU Haijin;WU Shan ;LIU Liyuan ;JIANG Dan;XU Tao;LAN Huatao;SHU Weitong;XU Guangxian;Department of Medical Examination,School of Medical Technology,Guangdong Medical University;

    Objective To construct a lentivirus-based expression plasmid and gene knockout plasmid of human interleukin(IL)-26 so as to lay a foundation of studying the function of IL-26 gene in cell signaling pathway and autophagy.Methods IL-26 gene sequence was amplified from human peripheral blood mononuclear cells by RT-PCR and cloned into pCDH-CMVMCS-EF1-copGFP eukaryotic expression vector to construct overexpression plasmid;Four knockout targets,Exon1sgRNA1,Exon1sgRNA2,Exon3sgRNA1 and Exon3sgRNA2,were designed based on the exon sequence of IL-26,and constructed into lentiCRISPRv2 vector by CRISPR/Cas9 technology to construct gene knockout plasmid. The overexpression plasmid and gene knockout plasmid were transiently transfected into HEK293T cells respectively,and the expression of IL-26 was verified by RT-qPCR and Western blot. In addition,amino acid sequence analysis,structure prediction and subcellular localization observation of IL-26 were performed.Results The results of restriction digestion,sequencing and bioinformatics analysis showed that IL-26 was 516 bp in length,encoding 171 amino acids. The IL-26 mRNA level and protein level of HEK293T cells transfected with IL-26 overexpression plasmid increased by 656. 789 times and 1. 978 times respectively with significant differences as compared with the normal control group(t = 17. 976 and 7. 859,P < 0. 000 1 and < 0. 001,respectively). With the transfection of 4 knockout targets Exon1sgRNA1,Exon1sgRNA2,Exon3sgRNA1 and Exon3sg-RNA2 into HEK293T cells,the expression of IL-26 decreased by 0. 930,0. 980,0. 523 3 and 0. 316 9 times,respectively,among which Exon3sgRNA2 significantly down-regulated the expression of IL-26(t = 7. 440,P < 0. 001). IL-26protein showed signal peptide structure and certain transmembrane function in the first 22 amino acids,which existed in cytoplasm.Conclusion IL-26overexpression and gene knockout plasmids were successfully constructed,which laid a foundation of the follow-up study of the function of IL-26.

    2024 02 v.37 [Abstract][OnlineView][Download 1382K]

  • Effect of follicular fluid-derived exosomal mi RNAs on follicular dysplasia mediated via glycolysis of granulosa cells in patients with polycystic ovary syndrome and its mechanism

    CAO Jianping;ZHANG Jianing;SHAN Huiquan;LIU Xuan;SU Jie;CUI Kuiqing;State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources,College Animal Science and Technology,Guangxi University;

    Objective To evaluate the effect of follicular fluid(FF)exosomal miRNAs on follicular dysplasia in patients with polycystic ovary syndrome(PCOS)mediated by glycolysis pathway of granulosa cells(GCs),and to explore the mechanism. Methods Three PCOS infertile patients and three non-PCOS infertile patients were recruited. The baseline hormone levels of the two groups were measured before ovulation induction. The bilateral FF was obtained by puncture after short-acting and long-term ovulation induction,and the exosomes were collected by ultracentrifugation and identified by transmission electron microscopy. The total exosomal RNA was extracted by Trizol method to construct the library,which was compared to the reference genome GRCh38 for statistical analysis after miRNA sequencing and quality control processing. Clustering Profiler R package was used to implement GO annotation analysis and KEGG pathway analysis of the differentially expressed genes(DEGs),and Omnipath software for miRNAs interaction analysis. A total of 16 miRNA were randomly selected and detected by qPCR to verify the accuracy of the miRNA sequencing results. Results Compared with the non-PCOS group,luteinizing hormone(LH),anti-Muerian hormone(AMH),testosterone and antral follicle counts in PCOS group increased significantly(t = 2. 479 ～ 9. 163,each P < 0. 05). The exosomes of FF in both groups showed the cup-shaped vesicles with clear edge and light staining in the center,with the diameters of 100 — 150 nm and intact structure,and the concentration was about 8 × 1010particles/mL. A total of 928 miRNAs were detected by miRNA sequencing. Compared with the non-PCOS group,59 differentially expressed miRNA(DEmiRNA)were screened out in exosomes of POCS group,of which 31 were up-regulated and 28 were down-regulated. The differential trend of gene expression detected by qPCR was highly similar to that of miRNA sequencing. In FF exosomes of PCOS patients,the glycolysis efficiency and apoptosis of GCs were significantly changed by miRNA regulating mRNA. PKM,PFKL and HK2 were the key target genes for miRNA to regulate GCs glycolysis,and SLC2A1 was the key target gene for miRNA to regulate GCs apoptosis. Conclusion The miRNAs in FF exosomes of PCOS patients can weaken the glycolysis of GCs while accelerate the apoptosis,thus reducing the production of ATP and lactic acid,resulting in follicular dysplasia.

    2024 02 v.37 [Abstract][OnlineView][Download 1341K]

  • Expression and immunogenicity of glycoprotein H of pseudorabies virus in mammalian cells

    YU Xiaohang;LIU Yining;DING Yanbin ;LUO Ye ;FANG Qi;ZHENG Jin ;YU Xinglong;College of Veterinary Medicine,Hunan Agricultural University;

    Objective To express glycoprotein H(gH)of pseudorabies virus(PRV)in mammalian cells and detect its immunogenicity.Methods The gH gene fragment of PRV-XiangA strain was amplified by PCR and inserted into mammalian cell expression vector pIRES-neo3 to construct recombinant expression plasmid pIRES-gH,which was transfected to HEK-293F cells and cultured in suspension for 5 d. The cell culture supernatant was identified by Western blot and purified by nickel ion chromatography column. The purified gH was emulsified with ISA 201 VG adjuvant to immunize 8 female ICR mice,and 8 mice in control group were immunized with the same amount of adjuvant,which was strengthened at 5 and 8weeks after the first dose respectively. The blood samples were collected at 4,7 and 10 weeks after the first dose and detected for the titer of specific antibody and neutralizing antibody in serum of mice;The mice were challenged with PRVXiangA strain(1. 5 × 104TCID50)by nasal drops 2 d after the third blood collection,and observed for the morbidity and mortality daily.Results The recombinant expression plasmid pIRES-gH was constructed correctly as identified by sequencing. The gH protein was successfully expressed and modified by glycosylation in mammalian cells with good reactivity,and about 625 μg purified protein was obtained under 100 mL culture volume. After three times of immunization,mice produced high level of specific antibody and showed the effect of neutralizing PRV,and the titer of neutralizing antibody reached 1∶256. In the challenge test,all the mice in control group became ill and died,while half of the mice in gH immunized group did not get sick with a survival rate of 50%.Conclusion PRV gH was successfully expressed in mammalian cells,and its immune protection was confirmed for the first time,which provided experimental basis for the further research and application of gH,and also provided a new idea for the development of PRV subunit vaccine.

    2024 02 v.37 [Abstract][OnlineView][Download 871K]

  • Isolation and identification of Lactobacillus paracasei and Bacillus subtilis in goose feces

    GAO Bingnan;YUE Xueqing;WANG Qiuju;CAO Yuhan;REN Yulong;SUN Yu;WEI Xiao;ZHANG Hao;College of Animal Science and Veterinary Medicine,Heilongjiang Bayi Land Agricultural University;

    Objective To isolate and identify Lactobacillus paracasei and Bacillus subtilis from feces of northern white geese.Methods Lactobacillus paracasei and Bacillus subtilis,numbered S00834-8 and IS00439-2 respectively,were isolated from feces of 30 52-week-old northern white geese and cultured in specific medium,and identified by 16S DNA sequencing. The two strains were cultured for different time duration(0,2,4,6,8,10,12,14,16,18,20,22,24,36 and 48 h),at different temperature(34,36,37,38 and 40 ℃)and pH(5. 5,6. 0,6. 5,7. 0,7. 5 and 8. 0). The growth characteristics of the strains were evaluated according to the A600values measured by UV-Vis spectrophotometer. At the same time,the acid and bile salt tolerance(pH 2. 0,2. 5,3. 0,3. 5 and 7. 0,bile salt concentration 0,0. 3%,0. 5%,1. 0% and 1. 5%)and the antibiotic tolerance were tested. Results The strains S00834-8 and IS00439-2 were Lactobacillus paracasei subspecies tolerance H and Bacillus subtilis KA9,respectively. The strain S00834-8 cultured under the optimum condition of the culture medium with pH 6. 0,at 37 ℃,for 12 h,had good bile resistance and gastric acid resistance,which was moderately susceptible to piperacillin,norfloxacin,amikacin,gentamicin and tetracycline,and susceptible to levofloxacin,streptomycin and chloramphenicol. The strain IS00439-2 best cultured in the culture medium with pH 7. 5,at 36 ℃,for 8 h,showed good bile resistance and gastric acid resistance,which was moderately sensitive to levofloxacin and tetracycline,while sensitive to piperacillin,norfloxacin,streptomycin and chloramphenicol. Conclusion Goose-derived Lactobacillus paracasei subspecies tolerance H and Bacillus subtilis KA9 have good acid resistance and bile salt resistance,which are expected to be candidate strains for goose-derived microecological preparations.

    2024 02 v.37 [Abstract][OnlineView][Download 994K]

  • Preparation and efficacy evaluation of polyclonal antibodies against serum albumin of five species

    ZHANG Jianmin;ZHOU Changming;LI Xiao;HU Yuchi;WANG Tiesong;ZHAO Xuan;Beijing Institute for Drug Control,NMPA Key Laboratory for Research and Evaluation of Generic Drugs,NMPA Key Laboratory of Safety Research and Evaluation of Innovative Drug;

    Objective To prepare polyclonal antibodies against the serum albumin of human,cattle,sheep,pig and horse,and evaluate their efficacy in the identification of human serum albumin(HSA). Methods The specific polypeptides of human,cattle,sheep,pig and horse serum albumin were prepared by bioinformatics and polypeptide synthesis method,which were coupled with keyhole limpet hemocyanin(KLH)to prepare the peptide antigen after the purity was identified by high performance liquid chromatography(HPLC). Male New Zealand white rabbits were immunized with polypeptide antigens of five species subcutaneously,with 2 for each kind of antigen. The antiserum was then obtained and purified by Protein A affinity chromatography to prepare the polyclonal antibody. The titers and specificity of the polyclonal antibodies were determined by ELISA and Western blot respectively,and the prepared five species of serum albumin were used to identify HSA products. Results The synthetic peptides of human,cattle,sheep,pig and horse serum albumin had a purity of over 91%,and the corresponding polyclonal antibodies all had the titer of 1∶160 000,which showed specific binding with the corresponding antigens and effectively identified the HSA products. Conclusion The polyclonal antibodies of human cattle,sheep,pig and horse serum albumin prepared in this study have good specificity and the preparation process is simple and rapid,suitable for the mass production,which lays a foundation of the development of HSA rapid identification kit.

    2024 02 v.37 [Abstract][OnlineView][Download 831K]

  • Effect of glutaminase 1 inhibitor on carbon tetrachloride-induced liver fibrosis in mice

    WANG Xiao;ZHANG Congcong ;ZHANG Yanhong ;HONG Shiyao ;DU Jie;Department of Nuclear Medicine,The First Hospital,Shanxi Medical University;

    Objective To investigate the effect of glutaminase 1(GLS1)specific inhibitor BPTES[bis-2-（5-phenylacetamido-1,3,4-thiadiazol-2-yl）ethyl sulfide]on the liver fibrosis in the mouse model of liver fibrosis induced by carbon tetrachloride(CCl4).Methods Male C57BL/6J mice were intraperitoneally injected with olive oil(control group),10%CCl4(10 μL/g,model group)or 10% CCl4(10 μL/g)+ BPTES(10 mg/kg,treatment group),with 10 mice in each group,two doses a week for four weeks to establish liver fibrosis model. Collagen deposition in mouse liver tissue was observed by Sirius red staining. The expression levels of actin alpha 2(Acta2),collagen typeⅠalpha 1(Col1a1)GLS1 and GLS1 protein were detected by qRT-PCR and immunohistochemical staining.Results Compared with the control group,the liver tissue of mice in the model group was generally enlarged,the surface was not smooth and granular,and the ratio of liver mass to tibia length significantly increased(t = 2. 979,P < 0. 05);The Sirius red positive area of collagen deposition increased signifi-cantly in the liver tissue of mice in the model group(t = 7. 661,P < 0. 01),the relative expression levels of Acta2 and Col1a1 significantly increased(t = 4. 335 and 5. 319,respectively,each P < 0. 01),and the mRNA and protein levels of GLS1 significantly increased(t = 5. 319 and 9. 725,respectively,each P < 0. 01). However,compared with the model group,the BPTES treatment group had a reduction in liver mass,a significant reduction in the Sirius red positive area of collagen deposition in liver tissue(t = 7. 427,P < 0. 01),and a significant reduction in the relative expressions of Atca2 and Col1a1(t = 3. 713 and 2. 628,respectively,each P < 0. 05).Conclusion Inhibition of GLS1activity can significantly improve the degree of liver fibrosis induced by CCl4,providing a new idea for the treatment of liver fibrosis.

    2024 02 v.37 [Abstract][OnlineView][Download 898K]

  • Protective effect of SRT1720 on acute liver injury induced by acetaminophen in mice and its mechanism

    ZOU Qiaoling;YUAN Qiulin ;SONG Yinhong;Hubei Key Laboratory of Tumor Microenvironment and Immunotherapy,China Three Gorges University;

    Objective To evaluate the protective effect of the activator of silent information regulator 2-related enzymes 1(SIRT1),SRT1720,on liver injury induced by acetaminophen(APAP)in mice and explore its mechanism. Methods Forty male C57BL/6J mice were randomly divided into normal control group,SRT1720 treatment group,APAP treatment group,and APAP + SRT1720 treatment group,with 10 mice in each group. Mice in SRT1720 and APAP + SRT1720 groups were given SRT1720(30 mg/kg body mass)by intragastric administration,while normal saline of equal volume was given by intragastric administration in control and APAP groups,once a day for 5 days;On the 6th day,mice in APAP and APAP + SRT1720 groups were injected i. p. with APAP(325 mg/kg body mass),while those in control and SRT1720 groups with equal volume of normal saline. After 24 hours,the peripheral blood was taken and the serum was separated,which were detected for the levels of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)by the corresponding kits;The liver tissue of mice was taken aseptically,observed for the pathological changes by HE staining,detected for the mRNA transcription levels of GRP78,PERK,eIF2 α,ATF4 and CHOP genes related to PERK-eIF2α-CHOP signaling pathway by RT-qPCR and detected for the relative expression levels of these corresponding proteins and Caspase12 protein by Western blot. Results Compared with normal control group,the serum ALT and AST levels of mice in APAP group significantly increased(t = 55. 21 and34. 29 respectively,each P < 0. 01);significant necrosis of hepatocytes was observed in liver tissue,the structure of hepatic lobules changed significantly,and the swelling and deformation of hepatocytes in some areas were serious;the mRNA transcription and relative protein expression levels of GRP78,PERK,eIFα,ATF4 and CHOP genes increased significantly(t = 9. 85～33. 89,each P < 0. 05)and the relative expression level of Caspase12 protein increased significantly(t = 11. 78,P < 0. 01). Compared with APAP group,the serum ALT and AST levels of mice in APAP + SRT1720 group decreased significantly(t = 42. 92 and 18. 02 respectively,each P < 0. 01);the degree of hepatocyte injury was obviously reduced and the number of swollen and deformed cells also significantly decreased;the mRNA transcription and relative protein expression levels of GRP78,PERK,eIF2α,ATF4 and CHOP decreased significantly(t = 6. 19～22. 43,each P < 0. 05)and the expression level of Caspase12 protein showed no significant decrease(t = 0. 34,P > 0. 05). Conclusion SRT1720improved APAP-induced liver injury in mice,possibly by inhibiting PERK-eIF2α-CHOP signaling pathway in endoplasmic reticulum stress(ERS).

    2024 02 v.37 [Abstract][OnlineView][Download 1021K]

  • Meta-analysis of safety and immunogenicity of DTaP-IPV-Hib-HepB hexavaccine

    LIU Jingwen;JIA Zheng ;LUO Xingxian ;FU Shuyong;YANG Yue ;XING Hua;School of Business Administration,Shenyang Pharmaceutical University;

    Objective To compare the differences of safety and immunogenicity of DTaP-IPV-Hib-HepB hexavaccine,DTaPIPV-Hib pentavaccine plus HepB single vaccine or DTaP-IPV-HepB pentavaccine plus Hib single vaccine,so as to provide a reference for the marketing and use of hexavaccine in China.Methods Randomized controlled trials(RCTs)of DTaP-IPVHib-HepB hexavaccine,DTaP triple vaccine,Hib,IPV and HepB vaccines published at home and abroad were searched.The safety and immunogenicity of the hexavaccine were evaluated by Meta-analysis using Revman 5.4.1 software.Results A total of 7 articles,8 RCTs and 3 429 subjects were included. Meta-analysis of safety showed that there was no significant difference in the incidence of injection site and systemic adverse reactions after vaccination with hexavaccine and pentavaccine plus single vaccine(P > 0. 05)except for induration at the inoculation site and crying. Meta-analysis of immunogenicity showed no significant difference in antibody indexes after vaccination with hexavaccine and pentavaccine plus single vaccine(P > 0. 05).Conclusion The safety and immunogenicity of DTaP-IPV-Hib-HepB hexavaccine in basic immunity was comparable to that of the control vaccine,and might be applied to infants and young children to prevent related diseases. However,due to the limitations of the quantity and quality of included studies,the above conclusions still depend on the further development of larger sample,multicenter and high-quality RCTs to verify.

    2024 02 v.37 [Abstract][OnlineView][Download 883K]

  • Analysis of HPV vaccine-related KAP and its influencing factors among resident women aged 16-45 years in Pudong New Area

    WANG Weiping;LIU Wenmin;YANG Dandan;JIE Junqin ;DU Li ;ZHOU Ping ;WEN Jinghai;Pudong Institute of Preventive Medicine of Fudan University,Pudong District Center for Disease Control and Prevention;

    Objective To analyze the related knowledge-attitude-practice(KAP)and the influencing factors of human papilloma virus(HPV)vaccine among 16 — 45 year old resident women in Pudong New Area.Methods Six of the 36streets(towns)in Pudong New Area were randomly selected as the survey site,the resident women aged 16 — 45 years were randomly selected excluding those with reading or comprehension disabilities and those with mental disorders,and a total of 1 022 valid questionnaires were collected through self-filling questionnaires to understand the KAP status of the HPV vaccine among the resident women in Pudong New Area. The relationship among KAP of HPV vaccine and its influencing factors were analyzed by univariate analysis and structural equation model.Results The overall awareness rate of HPV vaccine among 16-45 year old resident women in Pudong area was higher. Univariate analysis showed that marital status,educational level,employment status and household annual income were related to the awareness level of HPV vaccine（χ~2=12. 832,17. 636,16. 770 and 20. 030,respectively,each P < 0. 05）;Age,marital status,employment status and children′s status were correlated with HPV vaccination level（χ~2= 12. 382,25. 777,8. 830 and 20. 138,respectively,each P <0. 05）;HPV vaccine health education,HPV and HPV vaccine knowledge scores affected HPV vaccination status（χ~2=97. 561 and 68. 969,respectively,P < 0. 001）;Subjects' knowledge of cervical cancer was positively affected by knowledge of HPV infection(γ_(11)= 0. 756,P < 0. 001). Knowledge of cervical cancer not only positively affected subjects′ attitudes towards the efficacy of HPV vaccine(β_(21)= 0. 557,P < 0. 001),also had a direct effect on the HPV vaccination behavior of the subjects,showing a promoting effect(β_(31)= 0. 274,P = 0. 004). Subjects′ approval of the efficacy of HPV vaccine had a positive effect on their actual vaccination behavior(β_(32)= 0. 175,P = 0. 016).Conclusion The willingness of the 16 — 45 year old resident women in Pudong New Area to inoculate HPV vaccine was positive,but the actual vaccination rate was low. It is suggested to strengthen the HPV vaccine publicity while strengthening the education of cervical cancer and HPV infection,and consider the necessity of including the suitable age males in the scope of human vaccination.

    2024 02 v.37 [Abstract][OnlineView][Download 929K]

  • Development and verification of kinetic chromogenic quantitative detection method for endotoxin content in intermediate of component pertussis antigen

    ZHAO Ming;LIU Ting;MA Yu;WU Lijie;FU Hui;LI Shihui;Beijing Biological Products Institute Co.,Ltd.;

    Objective To develop a kinetic chromogenic quantitative method for the determination of endotoxin content in intermediate of pertussis antigen,and to verify the method so as to better control the quality of diphtheria,tetanus,and pertussis vaccine(DTP vaccine).Methods A kinetic chromogenic assay[Limulus Amebocyte Lysate(LAL)]was developed to detect the endotoxin content in the intermediate products of pertussis antigens after detoxification,and verified for the linearity,specificity,accuracy,reproducibility and intermediate precision. The quantitative detection results of kinetic chromogenic assay were compared with those of gel method.Results The absolute value of the linear correlation coefficient（|r|）of the kinetic chromogenic assay was more than 0. 99;under the maximum effective multiple dilution,the interference test recovery of the intermediate was within 50% — 200%,and pertussis toxin(PT)diluted to 10,100 and 1 000 times,filamentous hemagglutinin(FHA)diluted to 3 000,5 000 and 10 000 times,and pertussis adhesin(PRN)diluted to 50,75 and 100 times had no interference effect on the experiment after detoxification;the accuracy verification recovery rates of PT,FHA and PRN were 125%,110% and 99% respectively;and the CVs of reproducibility verification were 7. 21%,8. 31% and 5. 84%,and the CVs of intermediate precision verification were 6. 04%,16. 29% and 12. 23%,respectively.The bacterial endotoxin content of the three batches of pertussis antigen intermediates detected by kinetic chromogenic assay was consistent with that verified by gel method,both of which were less than the limit of bacterial endotoxin in the intermediates of pertussis antigen after detoxification.Conclusion The developed kinetic chromogenic assay has good linearity,specificity,accuracy and precision with accurate detection results,which can be used to detect the endotoxin content in intermediate products of component pertussis antigen after detoxification.

    2024 02 v.37 [Abstract][OnlineView][Download 826K]

  • Development and verification of a detection method for activity of neutralizing antibody in ELISA antibody positive serum of rats immunized with recombinant human interleukin-1 receptor antagonist

    PENG Binghuai;LIU Jinghui;LIU Yulin;SUN Zhongtao;SUN Jiwei;LIANG Guanqiao;YU Lu;Changchun Institute of Biological Products Co.,Ltd.;

    Objective To develop and verify a method for detecting the activity of neutralizing antibodies in ELISA antibody positive serum of rats immunized with recombinant human interleukin-1 receptor antagonist(rhIL-1Ra). Methods The SD rats were subcutaneously immunized with 3,20 and 100 mg/kg rhIL-1Ra injection respectively,10 rats in each group,half male and half female,twice a day at an interval of at least 4 h between each dose for 13 consecutive weeks. The blood samples were collected from the jugular vein of rats during the administration period and the recovery period. The serum samples were isolated and detected for the antibody titers by ELISA,and the samples positive for rhIL-1Ra antibody were purified by Protein A chromatographic column. Based on,D10G4·1 cells biological activity assay,a method for the detection of neutralizing antibody activity was developed and verified for the specificity,sensitivity and precision. The neutralizing antibody activity of rhIL-1Ra antibody positive serum determined by ELISA was detected by using the developed method.Results With the increase of doses,the serum antibody titers of rats in various dose groups gradually increased,and there were still antibodies in the recovery period,and the titer was still high. Rabbit anti-rhIL-1Ra monoclonal antibody showed obvious neutralizing effect on rIL-1Ra,while rabbit anti-rIFN-2b monoclonal antibody had no dose-effect relationship with rIL-1Ra. The sensitivity of the method was 171. 93 μg/mL;The CVs of precision verification were not more than 20%. The positive antibody sera detected by ELISA all had neutralizing effect on rhIL-1Ra injection,which was consistent with the results detected by ELISA. Conclusion The method developed in this study has good specificity and high sensitivity in the detection of serum neutralizing antibody activity in rats immunized with rhIL-1Ra,which can be used to detect the serum neutralizing antibody activity of animals with rhIL-1Ra repeated administration.

    2024 02 v.37 [Abstract][OnlineView][Download 858K]

  • Development and verification of an ELISA method for detection of process-specific residual host protein in biological preparation

    TIAN Yixiao;WANG Xinyue;DOU Xiangya;HAN Cuicui;GAO Ke;SHAO Dongyan;DUAN Suyang;BAI Yueqiu;ZHAO Wen;HUANG Qingsheng;Key Laboratory for Space Biosciences and Biotechnology,School of Life Sciences,Northwestern Polytechnical University;

    Objective To develop and verify a double-antibody sandwich ELISA method for the detection of process-specific E.coli residual protein in recombinant biological preparations.Methods Taking the production and purification process of glucagon-like peptide(GLP)expressed by E.coli as the specific process model,the same process was used to intercept the residual protein of empty E.coli(normal E.coli that does not express recombinant protein). One female New Zealand white rabbit and six female Kunming mice were immunized with the residual protein as the immunogen. Using the IgG antibody purified from rabbit immune serum as the coating antibody,mouse immune serum as the second sandwich antibody,and antimouse IgG-HRP as the enzyme-labeled secondary antibody,a double antibody sandwich ELISA method for process-specific residual protein of E.coli was established. The specificity,accuracy and precision of the method were verified,and the limit of detection(LOD)was determined. Simultaneously,the developed method and the commercial E.coli host protein residue detection kit were used to quantitatively determine the residual protein of purified GLP preparation.Results After a series of gradient dilution of process-specific residual protein with known concentration,the sensitivity of this ELISA method reached 338 pg/mL. No cross reaction occurred in the detection of CHO and yeast cell lysis protein by this method,the recoveries of samples with low,medium and high concentrations were all in the range of 80% — 120%,and the intra-assay and inter-assay CVs of the empty E.coli interception standard with low,medium and high concentrations were all less than15%. For the residual protein in GLP preparation,about 62% of the residual proteins were not detected by the commercial non-process-specific ELISA kit compared with the total amount of residual proteins detected by the developed method,and these residual proteins should be the process-specific residual proteins.Conclusion The double antibody sandwich ELISA method developed in this study has high sensitivity,strong specificity,good accuracy and precision for the detection of process-specific E.coli residual protein,which can meet the detection requirements that the residual protein is less than0. 01% — 0. 1% in biological preparations.

    2024 02 v.37 [Abstract][OnlineView][Download 848K]

  • Functions of HIV-1 Tat and its association with diseases

    LI Yuanhang;CHEN Ruixin;DU Juan;Institute of Virology and AIDS Research,First Hospital of Jilin University;

    Acquired immune deficiency syndrome,or AIDS,has been a major infectious disease that troubles the public health in a global scale. Human immunodeficiency virus type 1(HIV-1)is the causative reagent responsible for AIDS development. Even though the highly active anti-retroviral therapy(HAART,or the cocktail therapy)that has been widely applied could effectively suppress the infection and replication of HIV-1,the infected people suffer from other related diseases,such as the HIV-associated neurocognitive disorder(HAND). This paper mainly focused on the function of an important regulatory protein of HIV-1,trans-activator of transcription(Tat),and its correlation with HIV-1 replication and HAND development,so as to clarify the importance of developing anti-AIDS drugs targeting Tat protein.

    2024 02 v.37 [Abstract][OnlineView][Download 919K]

  • Research progress of chitosan and quaternized chitosan as vaccine adjuvant

    MA Shumin;XIANG Tingting;WO Enkang;School of Laboratory Medicine and Bioengineering,Hangzhou Medical College;

    Adjuvant is the key factor for many new vaccines to play a protective role. The addition of adjuvant can not only reduce the amount of antigen,but also increase the immunogenicity of its antigen,and stimulate the strong immune response of body. Chitosan,as the product of natural polysaccharide chitin removing part of acetyl group,has the characteristics of adhesion,permeability,biocompatibility and so on,and has been widely studied and applied as a vaccine adjuvant. As a novel adjuvant,it can not only help to induce cellular and humoral immunity,but also activate mucosal immunity. This review discussed the recent progress of chitosan and quaternized chitosan as vaccine adjuvants and the related mechanism.

    2024 02 v.37 [Abstract][OnlineView][Download 810K]

  • Research progress on autocrine and paracrine regulation of osteosarcoma cell growth

    DU Shaowen;LIU Xiang;YE Kaishan;Department of Orthopedics,Second Hospital of Lanzhou University,Key Laboratory of Bone and Joint Diseases of Gansu Province;

    Osteosarcoma(OS)is one of the most common malignant tumors in bone tissue,and its specific mechanism is not yet fully clear. Studies have shown that OS cells express different cytokines(CKs)and their receptors in the development stage of tumors,and enable them to grow autonomously and confer metastatic ability through autocrine and paracrine effects. In this regard,the most important CKs mainly includes insulin-like growth factor-1,transforming growth factor-β,chemokine 5 and interleukin-8,which can regulate the tumor microenvironment that is conducive to tumor growth,invasion and metastasis. This review summarizes the role and mode of action of CKs and their biological relevance to OS cells,hoping to provide effective new markers and therapeutic targets for clinical treatment of OS.

    2024 02 v.37 [Abstract][OnlineView][Download 822K]

  • Research progress on application of lactoferrin in clinical field

    MA Xiaomei;PENG Qiaofeng;ZHANG Haixia;MA Zhongren;Key Laboratory of Biotechnology and Bioengineering of State Ethnic Affairs Commission,Biomedical Research Center,Northwest Minzu University;

    Lactoferrin(LF),as a kind of iron-bound natural transferrin with wide functions,has become a research hotspot at home and abroad in recent years. Studies have shown that LF has a wide range of treatment,prevention and biological activity. This paper reviewed the clinical effects of LF in immune regulation,anti-tumor,regulation of obesity mechanism,antibacterial,anti-Alzheimer disease(AD)and bone regeneration mechanism in recent years,in order to provide a direction for the follow-up clinical application and research of LF.

    2024 02 v.37 [Abstract][OnlineView][Download 826K]

  • Research advances in monoclonal antibodies for treatment of drug resistance in inflammatory bowel disease

    GU Jiabo;WEI Ping;WANG Jun ;JIN Heiying;Anorectal Department,Lianyungang Hospital of Traditional Chinese Medicine;

    Inflammatory bowel disease(IBD)is a complex inflammatory disease mediated by immunity that is treated with the goal of maintaining disease remission and preventing recurrence. With the deep study of the molecular mechanism of the occurrence and development of IBD,many related target molecules have been found,and the monoclonal antibodies of corresponding targets have been used to treat the disease,while the drug resistance phenomenon generated during treatment has seriously affected the treatment effect. In this paper,monoclonal antibodies such as infliximab were reviewed for the treatment of IBD resistance,with a view to understanding its possible mechanisms and exploring effective treatments and preventive measures.

    2024 02 v.37 [Abstract][OnlineView][Download 834K]
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@2012 Editorial Department of "Chinese Journal of Biologicals"