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1.
Chinese Journal of Experimental and Clinical Virology ; (6): 212-216, 2019.
Article in Chinese | WPRIM | ID: wpr-804724

ABSTRACT

Objective@#To develop the chimeric antibodies against neuraminidase (NA) of H7N9 and to identify their biological activity and function.@*Methods@#The genes of variable regions of the light chain (VL) and heavy chain (VH) obtained by mouse hybridoma technology were cloned respectively into the expression VH and VL vectors bearing human-derived Cγ1, and Cκ1 and co-transfected into 293T cells. The chimeric antibodies were purified and their functions were investigated.@*Results@#Two chimeric antibodies, 1E2 and 3E3 against neuraminidase (NA) of H7N9 were obtained. Both antibodies recognized similar antigenic epitopes. MAb 1E2 and 3E3 could prevent the infectivity with H7N9 and H11N9 virus and reduce their size of viral plaque.@*Conclusions@#The chimeric antibodies specific for N9 could prevent the infection of N9 subtype influenza virus as well as the NAI-resistant mutants and could be a potential immunotherapy approach for H7N9 treatment.

2.
Journal of Preventive Medicine ; (12): 1233-1240, 2019.
Article in Chinese | WPRIM | ID: wpr-815967

ABSTRACT

Objective@#To systematically evaluate the effects of AIDS related health education on Chinese university students,and to provide reference for effective prevention and control of AIDS in universities. @*Methods@#Chinese and English databases including China National Knowledge Infrastructure,Wanfang Database,VIP Database,China Biology Medicine disc and PubMed were searched to find literature on the effects of AIDS health education on Chinese university students,which was published from June 30th of 2010 to June 30th of 2019. Rate difference(RD)was adopted as the effect index for the meta-analysis.@*Results@#A total of 173 articles were found out,28 articles of which were included. There were 48 037 study samples,with 23 020 students in the intervention group and 25 017 students in the control group. The results of the meta-analysis showed that compared with the control group,the awareness rates of knowledge about condom(RD=0.21,95%CI:0.11-0.32)and mosquito transmission(RD=0.34,95%CI:0.25-0.43),the rates of positive attitude to having class with HIV/AIDS(RD=0.21,95%CI:0.12-0.31)and being AIDS prevention volunteers(RD=0.14,95%CI:0.06-0.23),and the rate of avoiding unprotected sexual behaviors(RD=0.30,95%CI:0.22-0.38)in the intervention group were significantly improved. The results of subgroup analysis showed that the awareness rate of knowledge about mosquito transmission in the comprehensive intervention based on peer education group was significantly improved(RD=0.43,95%CI:0.32-0.53);the rate of positive attitude to having class with HIV/AIDS in medical students was significantly improved(RD=0.40,95%CI:0.35-0.46). No publication bias was found by Begger's test and Egger's test.@*Conclusion@#Through AIDS health education,the awareness of AIDS-related knowledge among Chinese university students has been obviously improved,while the intervention of attitude and behaviors still needs to be strengthened with the promotion of comprehensive intervention based on peer education.

3.
Chinese Critical Care Medicine ; (12): 1072-1077, 2019.
Article in Chinese | WPRIM | ID: wpr-791027

ABSTRACT

Objective To describe and compare the epidemiology of Sepsis-1 and Sepsis-3 in Yuetan Subdistrict of Beijing, and to estimate the incidence of Sepsis-3 in China. Methods A population-based cohort study was conducted. Through the database of Beijing Public Health Information System, the medical records of all adult residents hospitalized from July 1st, 2012 to June 30th, 2014 in Yuetan Subdistrict were reviewed. According to the clinical data of these patients, patients with Sepsis-1 and Sepsis-3 were enrolled in this analysis and the demographic characteristics of them were compared. Incidence and in-hospital mortality was calculated. Logistic regression method was used to analyze the risk factors of sepsis. Results Compared with non-septic patients with infections, patients with Sepsis-1 or Sepsis-3 were more likely to be male, older, had more comorbidities and lower body mass index (BMI), had more lower respiratory tract infections, more intra-abdominal infections and more blood flow infections, but had fewer urogenital tract infections, fewer upper respiratory tract infections, fewer gastrointestinal infections and fewer skin and soft tissue infections, and had pure prognosis with longer length of hospital stay [days, Sepsis-1 compared with non-Sepsis-1: 18 (10, 34) vs. 14 (9, 22), Sepsis-3 compared with non-Sepsis-3: 20 (11, 39) vs. 14 (9, 25)] and higher mortality [Sepsis-1 compared with non-Sepsis-1: 20.6% (353/1 716) vs. 2.5% (44/1 733), Sepsis-3 compared with non-Sepsis-3: 32.0% (299/935) vs. 3.9% (98/2 514), all P < 0.01]. Logistic regression analysis showed that male, elder (age ≥ 65 years old), low BMI, bed-rest state, and combined with cerebrovascular disease and hematological malignancies were risk factors for Sepsis-1 [all odds ratio (OR) > 1, all P < 0.05], while the male, elder (age ≥ 65 years old), low BMI, bed-rest state, and combined with cerebrovascular disease, rheumatic immune disease, malignant diseases of blood system and dementia were risk factors for Sepsis-3 (all OR > 1, all P < 0.05). Sepsis-3 was more common in males [OR = 1.19, 95% confidence interval (95%CI) was 1.01-1.40, P < 0.05] and elderly patients (age 65-84 years old: OR = 1.60, 95%CI was 1.28-1.99, P < 0.01; age ≥ 85 years old: OR = 1.76, 95%CI was 1.39-2.23, P < 0.01) as compared with Sepsis-1. After adjusted for gender and age, the standardized incidence of Sepsis-1 was 461 per 100 000 person-year, and that of Sepsis-3 was 236 per 100 000 person-year, with the standardized mortality of 79 per 100 000 person-year and 67 per 100 000 person-year, respectively, in Yuetan Subdistrict of Beijing. Corresponding to a speculative extrapolation of 4 856 532 new cases for Sepsis-1 and 2 487 949 new cases for Sepsis-3, there were 831 674 deaths and 700 437 deaths per year in China, respectively. Conclusions Male, elder, more comorbidities and low BMI were risk factors for sepsis. The standardized incidence of Sepsis-3 in Yuetan Subdistrict of Beijing was 236 per 100 000 person-year, and speculated there were 2.5 million new cases of Sepsis-3 per year, resulting in more than 700 000 deaths in China. According to the diagnostic criterion of Sepsis-3, 2.36 million new cases per year were reduced, and the mortality was increased by 11.4%, as compared with the criterion of Sepsis-1.

4.
Chinese Critical Care Medicine ; (12): 1072-1077, 2019.
Article in Chinese | WPRIM | ID: wpr-797521

ABSTRACT

Objective@#To describe and compare the epidemiology of Sepsis-1 and Sepsis-3 in Yuetan Subdistrict of Beijing, and to estimate the incidence of Sepsis-3 in China.@*Methods@#A population-based cohort study was conducted. Through the database of Beijing Public Health Information System, the medical records of all adult residents hospitalized from July 1st, 2012 to June 30th, 2014 in Yuetan Subdistrict were reviewed. According to the clinical data of these patients, patients with Sepsis-1 and Sepsis-3 were enrolled in this analysis and the demographic characteristics of them were compared. Incidence and in-hospital mortality was calculated. Logistic regression method was used to analyze the risk factors of sepsis.@*Results@#Compared with non-septic patients with infections, patients with Sepsis-1 or Sepsis-3 were more likely to be male, older, had more comorbidities and lower body mass index (BMI), had more lower respiratory tract infections, more intra-abdominal infections and more blood flow infections, but had fewer urogenital tract infections, fewer upper respiratory tract infections, fewer gastrointestinal infections and fewer skin and soft tissue infections, and had pure prognosis with longer length of hospital stay [days, Sepsis-1 compared with non-Sepsis-1: 18 (10, 34) vs. 14 (9, 22), Sepsis-3 compared with non-Sepsis-3: 20 (11, 39) vs. 14 (9, 25)] and higher mortality [Sepsis-1 compared with non-Sepsis-1: 20.6% (353/1 716) vs. 2.5% (44/1 733), Sepsis-3 compared with non-Sepsis-3: 32.0% (299/935) vs. 3.9% (98/2 514), all P < 0.01]. Logistic regression analysis showed that male, elder (age ≥ 65 years old), low BMI, bed-rest state, and combined with cerebrovascular disease and hematological malignancies were risk factors for Sepsis-1 [all odds ratio (OR) > 1, all P < 0.05], while the male, elder (age ≥ 65 years old), low BMI, bed-rest state, and combined with cerebrovascular disease, rheumatic immune disease, malignant diseases of blood system and dementia were risk factors for Sepsis-3 (all OR > 1, all P < 0.05). Sepsis-3 was more common in males [OR = 1.19, 95% confidence interval (95%CI) was 1.01-1.40, P < 0.05] and elderly patients (age 65-84 years old: OR = 1.60, 95%CI was 1.28-1.99, P < 0.01; age ≥ 85 years old: OR = 1.76, 95%CI was 1.39-2.23, P < 0.01) as compared with Sepsis-1. After adjusted for gender and age, the standardized incidence of Sepsis-1 was 461 per 100 000 person-year, and that of Sepsis-3 was 236 per 100 000 person-year, with the standardized mortality of 79 per 100 000 person-year and 67 per 100 000 person-year, respectively, in Yuetan Subdistrict of Beijing. Corresponding to a speculative extrapolation of 4 856 532 new cases for Sepsis-1 and 2 487 949 new cases for Sepsis-3, there were 831 674 deaths and 700 437 deaths per year in China, respectively.@*Conclusions@#Male, elder, more comorbidities and low BMI were risk factors for sepsis. The standardized incidence of Sepsis-3 in Yuetan Subdistrict of Beijing was 236 per 100 000 person-year, and speculated there were 2.5 million new cases of Sepsis-3 per year, resulting in more than 700 000 deaths in China. According to the diagnostic criterion of Sepsis-3, 2.36 million new cases per year were reduced, and the mortality was increased by 11.4%, as compared with the criterion of Sepsis-1.

5.
Chinese Journal of Experimental and Clinical Virology ; (6): 136-139, 2018.
Article in Chinese | WPRIM | ID: wpr-806035

ABSTRACT

Objective@#To isolate the cross-reactive antibodies against hemagglutinin of influenza virus and identify its biological function.@*Methods@#The antibodies gene reservoir of cross-reactive and H5N1 pseudotype particles neutralizing B cell circulating in peripheral blood of a human H5N1 case was recovered by in vitro B cell culture, screening, RT-PCR and expression vector cloning techniques. The Ab gene pairing was screened by transient transfection of human kidney 293T cells and detected using ELISA and neutralization test. The heterosubtypic antibodies were prepared and characterized.@*Results@#We discovered the VH1-2-based heterosubtypic antibodies from two B cell lineages could neutralize GX-H5N1 pseudotype particles and have broader binding with Group 1 (including H1, H5, H6 and H9) and H7 subtype.@*Conclusions@#Cross-reactive antibodies can be induced by H5N1 infection.

6.
Chinese Journal of Experimental and Clinical Virology ; (6): 66-70, 2017.
Article in Chinese | WPRIM | ID: wpr-807985

ABSTRACT

Objective@#To develop the monoclonal antibody (mAb) against neuraminidase of H7N9 subtype influenza A virus and identify its biological function.@*Methods@#Female 8 week-old BALB/c mice were immunized and the splenocytes of the mice were fused with Sp2/0 myeloma cells. Indirect ELISA was used to screen hybridoma and the positive clones were subject to be subcloned. Positive clones were identified and the monoclonal antibodies(mAbs) were obtained by purifying the ascetic fluid of mice injected with the hybridoma. The NA-binding as well as neuraminidase-inhibition activity of these mAbs were determined.@*Results@#Three mAbs against neuraminidase of H7N9 subtype influenza A virus, 1G8, 3C4 and 4E8, were obtained. They demonstrated different epitop-recognizing. 3C4 and 4E8 exhibited neuraminidase inhibitory activity, with a IC50 of 1.45 μg/ml and 8.65 μg/ml, respectively.@*Conclusions@#The results suggested that mAbs specific to neuraminidase of H7N9 subtype influenza A virus were developed, providing an useful tool in control and preventing the novel H7N9 influenza A virus.

7.
Chinese Journal of Microbiology and Immunology ; (12): 16-19, 2016.
Article in Chinese | WPRIM | ID: wpr-488859

ABSTRACT

Objective To construct a reverse genetic platform for influenza B virus and to rescue influenza B virus.Methods Eight plasmids carrying the gene segments of B/Florida/4/2006 virus were constructed by using the bidirectional promoter vector pHW2000.293T cells were co-cultured with MadinDarby canine kidney (MDCK) cells and then transfected with the eight plasmids.The supernatants of cell culture and cell debris were collected after transfection and then injected into embryonated chicken eggs and MDCK cells for rescuing the influenza B virus strains.Results This reverse genetic system could be used for the preparation of reassortant influenza B virus strains.The titers of hemagglutination units of the rescued virus achieved 128-256/50μl.Most of the reassortant virus particles were spherical under electron microscope.Conclusion The pHW2000 reverse genetic system could be used for the rescue of influenza B virus.Moreover,it could also be used for the construction of influenza B virus with specific mutations for further in vestigation on the characteristics of influenza B virus and the construction of vaccine strain.

8.
Chinese Journal of Microbiology and Immunology ; (12): 98-102, 2016.
Article in Chinese | WPRIM | ID: wpr-487501

ABSTRACT

Objective To express the receptor binding domain (RBD) protein of the Middle East respiratory syndrome coronavirus (MERS-CoV) and to characterize the antigenicity of the purified recombi-nant protein. Methods The codon-optimized gene encoding the RBD protein of MERS-CoV was synthesized and then cloned into the pET30a ( +) vector to construct the recombinant expression plasmid. The trans-formed E. coli BL21 (DE3) strains carrying expression plasmid were induced by IPTG under different condi-tions. The expressed products were purified by using nickel affinity chromatography and further analyzed by SDS-PAGE and Western blot assay. Indirect ELISA was performed to analyze the antigenicity and specificity of RBD proteins expressed in prokaryotic expression systems in human serological test. Results The recom-binant RBD proteins were mainly expressed as conclusion body in an optimal induction condition of 37℃ and 0. 5 mmol/ L IPTG for 4 h. The high purified recombinant RBD proteins were obtained through denaturation and renaturation with a relative molecular mass of about 29×103 . Results of the Western blot assay showed that the recombinant RBD proteins could have specific reaction with the serum samples collected form mice with MERS-CoV infection. Indirect ELISA revealed that the RBD proteins expressed in the prokaryotic ex-pression system showed better sensitivity and specificity in the detection of antibodies against MERS-CoV in human serum samples. Conclusion This study reported the prokaryotic expression and purification of RBD protein of MERS-CoV for the first time, which might pave the way for further investigation on immunological detection of MERS-CoV and development of vaccines against MERS-CoV infection.

9.
Chinese Journal of Virology ; (6): 524-529, 2015.
Article in Chinese | WPRIM | ID: wpr-296253

ABSTRACT

Hemagglutinin (HA) contains a head domain with a high degree of variability and a relatively conserved stem region. HA is the major viral antigen on the surface of the influenza virus. To define the biologic activities of chimeric HA bearing different head domains and stem regions or their potential use, a HA chimeric gene containing the head domain of the H7 subtype virus and stem region of the H3 subtype virus was modified and expressed using a baculovirus expression vector. Then, the secreted protein was purified and its biologic activities characterized. Approximately 1.4 mg/mL cH7/3 HA could be obtained, and its molecular weight was ≈ 70 kD. The trimer form of cH7/3 protein had hemagglutination activity and could be recognized by specific antibodies. The method described here can be used for further studies on the screening of HA stem-reactive antibodies or the development of vaccines with conserved epitopes.


Subject(s)
Humans , Antibodies, Viral , Allergy and Immunology , Baculoviridae , Genetics , Metabolism , Gene Expression , Genetic Vectors , Genetics , Metabolism , Hemagglutination , Hemagglutinin Glycoproteins, Influenza Virus , Genetics , Allergy and Immunology , Influenza Vaccines , Genetics , Allergy and Immunology , Influenza, Human , Virology
10.
Chinese Journal of Microbiology and Immunology ; (12): 668-672, 2014.
Article in Chinese | WPRIM | ID: wpr-671871

ABSTRACT

Objective To develop an effective and broad immune protective vaccination strategy by using DNA and recombinant vaccinia-based H5N1 vaccines.Methods BALB/c mice were immunized with various prime-boost regimens by using different DNA ( pIRES-based or pVRC-based) and recombinant vaccinia (Tiantan strain, rTTV)-based H5N1 vaccines expressing multivalent antigens (HA, NA, M1 and M2).The differences of immunity induced by two DNA vaccines were compared between intradermal electro -poration (IDE) and intramuscular electroporation (IME) deliveries.Immune responses were analyzed by hemagglutination inhibition( HAI) assay, neuraminidase ( NA)-specific antibody measured by ELISA , mi-croneutralization assay and IFN-γELISPOT assay .Results High levels of humoral immunity and T cell re -sponses were induced in mice primed with DNA-based vaccine than those primed with rTTVb-ased vaccine . DNA priming by IDE resulted in higher levels of neutralizing antibody in mice than those by IME delivery . Higher levels of HAI and anti-NA antibodies as well as NA-specific T cell responses were induced by pVRC-based DNA prime than those by pIRES-based DNA prime .HA-specific T cell responses were also enhanced in mice primed with pVRC-based DNA than those primed with pIRES-based DNA by IME .Conclusion The prime-boost strategies by using DNA-based vaccine in combination with rTTV-based H5N1 vaccine could induce humoral and T cell responses in mice against multi-antigens .Immunities induced by vaccines in com-bination might be modulated by various prime regimes .The study provided references for the further develop-ment of novel H5N1 vaccine and the optimization of immunization programs of combined multi-antigen vac-cine candidates .

11.
Chinese Journal of Laboratory Medicine ; (12): 743-747, 2013.
Article in Chinese | WPRIM | ID: wpr-437820

ABSTRACT

Objective To predict the effectiveness of gentamicin combined with trisodium citrate for locking solution in preventing catheter-related bloodstream infection.Methods This is a retrospective study,ten kinds of common pathogens in catheter-related bloodstream infections were selected from medical intensive care unit (n =9) and cardiac care unit (n =1) between July 2011 and July 2012,and the diffusion method was used to analyze the inhibitory effect of gentamicin combined with trisodium citrate against tested pathogens.Results When trisodium citrate was tested alone,a certain antimicrobial activity could be detected from 10% trisodium citrate,only Staphylococcus aureus and Staphylococci haemolyticus were inhibited,and then Enterococcus faecium was inhibited by 30% trisodium citrate.Compared with trisodium citrate,gentamicin showed stronger antimicrobial activity.Staphylococcus aureus,Staphylococci haemolyticus,Enterococcus faecium and Escherichia coli could be inhibited by 0.32 g/L gentamicin and the antimicrobial activity was enhanced by increasing concentrations.The maximum activity was in 5 g/L gentamicin with 7-17 mm diameters.However,the antimicrobial activity of gentamicin didn't increase when combined with trisodium citrate,especially in 4% trisodium citrate,the Staphylococci haemolyticus was not inhibited by all of the combinations with any concentration of gentamicin.In 0.32 g/L gentamicin,only the combination with 50% trisodium citrate could inhibit the four kinds of pathogens (Staphylococcus aureus,Staphylococci haemolyticus,Enterococcusfaecium and Escherichia coli) with 8-21 mm diameters,while in 1 g/L,2 g/L,and 5 g/L gentamicin,these pathogens could be inhibited by the combination with only 10% trisodium citrate,and the diameters were 5-24 mm,12-27 mm,and 17-28 mm,respectively.Conclusion Based on the data,gentamicin combined with trisodium citrate lock solution may play an important role in preventing catheter-related bloodstream infections caused by Gram-positive bacteria and Escherichia coli.

12.
Biomedical and Environmental Sciences ; (12): 530-536, 2011.
Article in English | WPRIM | ID: wpr-235600

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the pathogenesis and immunogenicity of H9N2 influenza virus A/Guangzhou/333/99 (a reassortant of G1 and G9 viruses isolated from a female patient in 1999) in a mouse model of infection.</p><p><b>METHODS</b>Mice were infected with increasing virus titers. Viral load in the lungs and trachea was determined by EID50 assay. Pulmonary histopathology was assessed by hematoxylin-eosin staining. Anti-HI antibody titers and T-cell responses to viral HA were determined by ELISPOT and confirmed by flow cytometry.</p><p><b>RESULTS</b>Mice presented a mild syndrome after intranasal infection with A/Guangzhou/333/99 (H9N2) influenza virus. Virus was detected in the trachea and lungs of mice harvested on days 3, 6, and 9 post-infection. A T-cell response to viral HA was detected on day 6 and H9 HA-specific CD(4+) T-cells predominated. Seroconversion was detected after 14 days and antibody persisted for at least 28 weeks.</p><p><b>CONCLUSION</b>Our results suggest that H9N2 (A/Guangzhou/333/99) can replicate in the murine respiratory tract without prior adaptation, and both humoral and cell-mediated immunity play an important role in the immune response.</p>


Subject(s)
Animals , Dogs , Female , Humans , Infant , Mice , CD4-Positive T-Lymphocytes , Allergy and Immunology , CD8-Positive T-Lymphocytes , Allergy and Immunology , Cell Line , Enzyme-Linked Immunospot Assay , Hemagglutination Inhibition Tests , Hemagglutinins, Viral , Allergy and Immunology , Influenza A Virus, H9N2 Subtype , Allergy and Immunology , Virulence , Interferon-gamma , Allergy and Immunology , Lung , Virology , Lymphocytes , Allergy and Immunology , Mice, Inbred BALB C , Orthomyxoviridae Infections , Allergy and Immunology , Virology , Spleen , Allergy and Immunology , Trachea , Virology , Viral Load , Virulence
13.
Chinese Journal of Microbiology and Immunology ; (12): 546-550, 2010.
Article in Chinese | WPRIM | ID: wpr-382788

ABSTRACT

Objective To develop an effective and broad immune protective H5N1 vaccine.Methods We first developed two recombinant vaccinia ( Tiantan strain) virus ( rTTV ) based H5N1 vaccines, which consisted of bicistron expressing the hemagglutinin(HA) and matrix protein 2(M2), or bicistron expressing the neuraminidase(NA) and matrix protein 1 (M1). The expression of H5N1 protein in rTTVs was confirmed. We immunized the BALB/c mice twice with two kind of dose ( 104 PFU, 107 PFU)using different combination. Subsequently, we assessed the humoral and cellular immune response in vaccinated mice. Results Our data showed that rTTV-based H5N1 vaccine induced rapidly robust HA- and NAspecific antibody level and IFN-γ secreting form cell(SFC) with either single dose of 107 PFU or twice dose of 104 PFU or 107 PFU. We also detected significant neutralizing antibody and matrix-specific immune response. In addition, we found that immunization with two kind of rTTV-based H5N1 vaccines induced much high level of M2-specific antibody than that with single of rTTV-based H5N1 vaccine. Conclusion rTTVbased H5N1 vaccines in this study elicited board array of immunity and our study offers a promising alternative H5N1 vaccine candidates with favorable potential to prevent various H5N1 pandemic.

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