1.Molecular epidemiology of noroviruses from infantile diarrhea at Suzhou and Nanjing area in Jiangsu province in 2010
Jianguang FU ; Bin WU ; Hong JI ; Liang LI ; Xian QI ; Yuanfang QIN ; Shenjiao WANG ; Fei DENG ; Zhifeng LI ; Fenyang TANG ; Rongqiang ZU ; Changjun BAO
Chinese Journal of Microbiology and Immunology 2011;31(11):989-993
ObjectiveTo investigate norovirus infection status and indentify its epidemiological characteristics and genotype distribution in infantile viral diarrhea in Jiangsu.MethodsFour hundred and ninety-eight fecal specimens of infantile virus diarrhea cases were collected from Suzhou Children's hospital and Nanjing Children's hospital in 2010.Norovirus genegroup were detected by real-time RT-PCR,and genetype were determined by sequence analysis.Results Among all fecal specimens,2 (0.4%) cases were positive for norovirus G Ⅰ,and 190 (38%) cases for G Ⅱ.Nucleotide sequence analysis revealed that in the 2 samples for G Ⅰ,one strain was G Ⅰ 1 and another was G Ⅰ 3.Twenty-one strains were belonged to G Ⅱ 4 and 2 strains were G Ⅱ 3 in the 23 samples for G Ⅱ.ConclusionAs one of the most important pathogens causing infantile viral diarrhea in Jiangsu province,subtype G Ⅱ 4 was the main epidemic strain of norovirus,meanwhile other genotypes also existed.
2.Knowledge,beliefs and behaviors related to HIV/AIDS among freshmen in Jiangsu colleges
CHEN Yuanfang, XIAO Wenjing, LU Jing, QIU Tao, LI Jianjun, FU Gengfeng
Chinese Journal of School Health 2020;41(1):47-50
Objective:
To understand the knowledge, attitude and behavior of HIV/AIDS prevention and treatment of freshmen in Jiangsu Colleges, so as to provide evidence for carrying out HIV/AIDS prevention and treatment propaganda among young college students.
Methods:
A total of 2 538 freshmen in 8 colleges in Jiangsu province were surveyed by cluster sampling, and anonymous self-reported questionnaire survey was carried out. Valid questionnaires were collected and analyzed.
Results:
Among 2 538 students valid, the total awareness rate of the eight items of new HIV/AIDS basic knowledge which provided to the public by the State (referred to the new eight items) was 85.74%, the awareness rate of boys was higher than that of girls’(88.21%, 84.34%)(P<0.05), the rate of college students of science was higher that of those of arts(87.31%, 83.67%)(P<0.05). the general awareness rate of other knowledge was 72.14%, and the rate of boys was higher than that of girls(75.00%, 70.53%)(P<0.05). As for "whether eating with the AIDS-affected patients may be affected with AIDS/HIV", boys and students of science were more rational(58.3%, 55.1%; 50.4%, 50.9%)(P<0.05). 50.08% students approved of having sexual behavior before marriage, and the rate of boys was higher than that of girls(69.00%, 39.40%)(P<0.01). 71.24% of the students thought that condoms should be used consistently in sex behaviors, and the proportion of female students (80.76%) was much higher than that of male students(54.37%), and the rate of the students of science was higher than that of students of arts(73.36%, 69.63%)(P<0.05). 6.03% students had received consult and test of AIDS. 40.34% of the untested students were willing to receive the test but didn’t know where to go. 1.34% students were afraid of receiving the test because of fear. The most popular ways of HIV/AIDS publicity among students are micro films(76.32%), lectures(66.67%) and peer education(60.80%).
Conclusion
It is suggested to publicize HIV/AIDS knowledge among students according to different gender and subject characteristics and combining multimedia, with emphasis on strengthening the publicity of HIV/AIDS epidemiology,treatment and related information, and attaching importance to sexual health education among young students.
3.Prokaryotic expression and bioreactivity analysis of a major epitope region of 2C with 3AB within non-structural protein of foot-and-mouth disease virus.
Xiaoli ZHANG ; Meina TIAN ; Zengjun LU ; Yuanfang FU ; Xiaojun MA ; Zaixin LIU ; Qingge XIE
Chinese Journal of Biotechnology 2009;25(1):10-15
In recent years, the potential value of nonstructural protein (NSP) 2C was well documented for distinguishing foot-and-mouth disease virus (FMDV) in infected animals and vaccinated animals. In order to develop a more sensitive approach to detect natural infected FMDV while there is no interact with vaccinated FMDV, we incorporated a major epitope region of 2C with whole 3AB coding region within NSP and expressed in Escherichia coli. We got a 47.6 kD fusion protein named 2C'3AB. The product showed a specific reactivity with FMDV from serum of infected animal by using Western blotting analysis. This suggests that this protein could be applied to distinguish infected FMDV and vaccinated FMDV. We further compared 2C'3AB protein with 3ABC fusion protein, another available protein used for detecting infected FMDV, using indirect ELISA assay. The results showed that 2C'3AB-ELISA had higher sensitivity than that of 3ABC-ELISA for distinguishing infected FMDV and vaccinated FMDV of sera from epidemic region. Therefore, this recombinant protein 2C'3AB is a good candidate protein to develop more sensitive method to differentiate infected FMDV and vaccinated FMDV from vaccinated animals. This finding will increase our capability to check the infectious virus carrier and finally improve FMDV infection control.
Animals
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Antibody Specificity
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Carrier Proteins
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genetics
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immunology
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metabolism
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Epitopes
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immunology
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Escherichia coli
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genetics
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metabolism
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Foot-and-Mouth Disease Virus
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genetics
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Recombinant Fusion Proteins
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genetics
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immunology
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metabolism
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Viral Nonstructural Proteins
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genetics
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immunology
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metabolism
4.Enhanced immune response of a novel T-cell immunogen in vaccine for foot-and-mouth disease.
Qing ZHAO ; Pu SUN ; Zaixin LIU ; Pinghua LI ; Huifang BAO ; Yimei CAO ; Xingwen BAI ; Yuanfang FU ; Zengjun LU ; Dong LI
Chinese Journal of Biotechnology 2011;27(9):1281-1291
We investigated the enhanced immune response of a recombinant T cell immunogen as an effective cellular immune adjuvant. The T cell immunogen named TI contained several T cell epitopes from the VP1, VP4, 3A and 3D proteins of foot-and-mouth disease virus (FMDV) and two pan-T helper (T(H)) cell sites to broaden the immunogenicity of the protein. Meanwhile, another fusion protein named OA-VP1 was expressed in bacteria, which contained two VP1 proteins of O and Asia1 type FMDV. Mice were vaccinated with commercially inactivated vaccine or OA-VP1 protein with or without the TI immunogen. The results show that mice inoculated with inactivated vaccine or OA-VP1 protein supplemented with TI immunogen produced significantly higher level of neutralizing antibodies (P < 0.01 or P < 0.05) than the mice only inoculated with inactivated vaccine or OA-VP1 protein by microneutralization assay. An obvious increase in T cell number by flow cytometric analysis and significantly higher concentration of IFN-gamma secreted in culture media of spleen lymphocytes were observed in groups supplemented with TI immunogen (P < 0.01). TI immunogen was an effective stimulator for humoral and cellular immunity and could help improve the immunogenicity of inactivated vaccine or protein subunit vaccine.
Adjuvants, Immunologic
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pharmacology
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Animals
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Capsid Proteins
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genetics
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immunology
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Epitopes, T-Lymphocyte
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genetics
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immunology
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Foot-and-Mouth Disease
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immunology
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prevention & control
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virology
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Foot-and-Mouth Disease Virus
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immunology
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Immunization
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Mice
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Viral Vaccines
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genetics
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immunology
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pharmacology
5.Clinical investigation of Q. Flex for improvement of PET/CT image quality and quantitative accuracy of pulmonary nodules
Dong DAI ; Jianjing LIU ; Di LU ; Guoqing SUI ; Yaya WANG ; Xueyao LIU ; Yuanfang YUE ; Zhen YANG ; Qing YANG ; Jie FU ; Wengui XU ; Ziyang WANG
Chinese Journal of Nuclear Medicine and Molecular Imaging 2024;44(2):98-103
Objective:To compare the imaging quality and metabolic quantitative parameters of pulmonary nodules between Q. Flex whole information five-dimensional (5D) and conventional three-dimensional (3D) PET/CT imaging for clinical evaluation.Methods:Fifty-four patients (30 males, 24 females, age: 60(42, 75) years; 78 solid pulmonary nodules (maximum diameter≤3 cm) with abnormal uptake of 18F-FDG) from Tianjin Cancer Hospital Airport Hospital between June 2022 and August 2022 were enrolled in this retrospective study. All patients underwent 5D scanning and 3D, 5D reconstruction. Image quality scores, signal-to-noise ratio (SNR), SUV max, SUV mean and metabolic tumor volume (MTV) of pulmonary nodules of 5D group and 3D group were evaluated and compared with χ2 test and Wilcoxon signed rank test. Correlation of quantitative parameters between 2 groups were analyzed by using Spearman rank correlation analysis. Results:Thirty-five of 78(45%) pulmonary nodules with image quality score≥4 were found in 5D group, which were more than those in 3D group (22/78(28%); χ2=4.67, P=0.031). Meanwhile, SNR, SUV max, SUV mean, and MTV were significantly positively correlated between the 2 groups ( rs values: 0.86, 0.86, 0.85, and 0.95, all P<0.001). SNR, SUV max and SUV mean of pulmonary nodules in 5D group were significantly higher than those in 3D group, which were 37.46(18.42, 62.00) vs 32.72(16.97, 54.76) ( z=-4.07, P<0.001), 9.71(5.48, 13.82) vs 8.96(4.82, 12.63) ( z=-3.05, P<0.001) and 6.30(3.39, 8.94) vs 5.61(2.99, 7.63)( z=-4.07, P<0.001) respectively. MTV of pulmonary nodules in 5D group was significantly lower than that in 3D group, which was 1.72(0.66, 2.74) cm 3vs 1.98(1.06, 4.63) cm 3 ( z=-7.13, P<0.001). Quantitative parameters of lower lung field and nodules with maximum diameters of >10 mm and ≤20 mm based on 5D scanning changed most significantly compared with those based on 3D scanning ( z values: from -5.23 to -2.48, all P<0.05). Conclusion:Q. Flex 5D PET significantly improves the quantitative accuracy of SUV and MTV of pulmonary nodules, and the improvement of image quality is substantial without increasing the radiation dose, which has clinical practical value.
6.Development of a sandwich ELISA for detecting 3AB non-structural protein of foot-and-mouth disease virus.
Yuanfang FU ; Wei HE ; Pu SUN ; Lin YANG ; Huifang BAO ; Yimei CAO ; Xingwen BAI ; Pinghua LI ; Dong LI ; Yingli CHEN ; Lei LIU ; Zengjun LU ; Zaixin LIU
Chinese Journal of Biotechnology 2020;36(11):2357-2366
Antigenic purity is important for quality control of the foot-and-mouth (FMD) whole virus inactivated vaccine. The recommended method for evaluation the antigenic purity of FMD vaccine is to check the serum conversion to non-structural protein (NSP) 3AB antibody after 2 to 3 times inoculation of animals with inactivated vaccine. In this study, we developed a quantitative ELISA to detect the amount of residual 3AB in vaccine antigen, to provide a reference to evaluate the antigenic purity of FMD vaccine. Monoclonal antibody (Mab) of NSP 3A and HRP-conjugated Mab of NSP 3B were used to establish a sandwich ELISA to quantify the NSP 3AB in vaccine antigen of FMD. Purified NSP 3AB expressed in Escherichia coli was serially diluted and detected to draw the standard curve. The detectable limit was determined to be the lowest concentration of standard where the ratio of its OD value to OD blank well was not less than 2.0. Results: The OD value was linearly corelated with the concentration of 3AB protein within the range between 4.7 and 600 ng/mL. The correlation coefficient R² is greater than 0.99, and the lowest detectable limit is 4.7 ng/mL. The amount of 3AB protein in non-purified inactivated virus antigen was detected between 9.3 and 200 ng/mL depending on the 12 different virus strains, whereas the amount of 3AB in purified virus antigen was below the lowest detectable limit. The amount of 3AB in 9 batches of commercial FMD vaccine antigens was between 9.0 and 74 ng/mL, whereas it was below the detectable limit in other 24 batches of commercial vaccine antigens. Conclusion: the sandwich ELISA established in this study is specific and sensitive to detect the content of 3AB protein in vaccine antigen of FMD, which will be a useful method for evaluation of the antigenic purity and quality control of FMD inactivated vaccine.
Animals
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Antibodies, Viral
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Enzyme-Linked Immunosorbent Assay
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Foot-and-Mouth Disease/prevention & control*
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Foot-and-Mouth Disease Virus
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Viral Nonstructural Proteins/genetics*
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Viral Vaccines
7.A method for immortalizing swine monoclonal B cells secreting anti-PRRSV antibodies.
Jian WANG ; Jing ZHANG ; Kun LI ; Pu SUN ; Guoxiu LI ; Jiaoyang LI ; Yimei CAO ; Zhixun ZHAO ; Hong YUAN ; Yuanfang FU ; Pinghua LI ; Dong LI ; Zaixin LIU ; Zengjun LU
Chinese Journal of Biotechnology 2022;38(8):2872-2882
Porcine reproductive and respiratory syndrome (PRRS) is a highly contagious disease caused by porcine reproductive and respiratory syndrome virus (PRRSV), which causes great economic losses. At the moment, no effective neutralizing antibody is available for scientific research and treatment. Therefore, developing a method for screening the neutralizing monoclonal antibodies is of great significance for the prevention and treatment of PRRSV and the screening of antigen sites. Monoclonal antibodies have been widely used in the treatment and diagnosis of many human and animal diseases. Therefore, screening effective neutralizing antibodies for different pathogens is an urgent task. Among the methods for monoclonal antibody screening, B cell immortalization is an effective method to obtain neutralizing monoclonal antibody. Specifically, in this study, the bcl-6 and bcl-xl genes were connected by f2a and then the yielded product was ligated to a vector for retrovirus packaging. The swine lymphocytes immunized with PRRSV were infected the yielded mature viruses and cultured in the complete medium containing CD40L and IL21 cytokines. Then, CD21 was used as the marker to screen B cells with the magnetic bead method. Finally, monoclonal B cells were obtained and the secretion of antibodies was tested. The results showed that the plasmid, either being transfected alone or with the packaged plasmids, could be expressed, and that the packaged retrovirus could infect the cells. Moreover, the infected lymphocytes secreted antibodies, so did the screened B cells. Therefore, the method for screening monoclonal antibody against PRRSV was successfully established.
Animals
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Antibodies, Monoclonal
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Antibodies, Neutralizing
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Antibodies, Viral
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Humans
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Porcine Reproductive and Respiratory Syndrome/prevention & control*
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Porcine respiratory and reproductive syndrome virus/genetics*
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Swine