1.Study on the correlation between abnormal menstrual cycle and intestinal microbiome in female rhesus monkeys
Minxue XIE ; Chen ZHAO ; Yuchen YAN ; Zhenghua PEN ; Jiaochun LI ; Yinzhen TAN ; Xuefu WANG ; Chaowu ZHANG ; Wu YANG ; Yuan ZHAO
China Modern Doctor 2024;62(17):1-6,12
Objective Using healthy female reproductive-age rhesus macaques as the research subjects,we explored the correlation between menstrual cycle abnormalities and gut microbiota composition by using 16S rRNA metagenomic sequencing.Methods Twenty-seven healthy female rhesus macaques were divided into regular menstrual and irregular menstrual groups.Fecal samples were collected at follicular phase(FP),ovulation phase(OP)and luteal phase(LP)of the two groups.The structure and diversity of bacterial flora in different physiological periods were analyzed and compared between the two groups.Results At the phylum level,Firmicutes,Bacteroidetes,and Proteobacteria dominated the sample flora in the follicular,luteal,and ovulatory phases of the rhesus macaques in both the regular and irregular groups,with a combined percentage of more than 98% .At the genus level,the genus Prevotella_9,Ruminococcaceae_UCG-002,Lactobacillus,Prevotella_2,Phascolarctobacterium,Ruminococcaceae_UCG-005,Streptococcus,Blautia,Prevotellaceae_NK3B31_group,Rikenellaceae_RC9_gut_group were dominant.In the luteal phase the percentage of Firmicutes was higher in the regular group than in the irregular group,while the opposite was true for Bacteroidetes.Spirochaetes were higher in the regular group than in the irregular group at all 3 stages(P<0.05).Conclusion There were some differences in intestinal microbial composition between the two groups of macaques with regular and irregular menstrual cycles,which provided some reference for the study of intestinal bacteria and ovulation disorders.
2.Construction and identification of multiple epitope antigens of norovirus based on bioinformatics
Xue DU ; Yinzhen ZHAO ; Yiqing ZHANG ; Xiaojun WANG ; Xudong WANG ; Lanying GUO ; Yunlong WANG
Chinese Journal of Immunology 2024;40(11):2391-2398
Objective:To design a multi-epitope antigen of norovirus(NoV)based on bioinformatics technology and to pre-pare and characterize it.Methods:Bioinformatics methods were used to construct and analyze the NoV multi-epitope antigen NoV-ZH.Recombinant proteins were prepared and characterized by prokaryotic expression system,and monoclonal antibodies were prepared by animal immunization and hybridoma technology,and initially applied in colloidal gold platform.Results:The designed multi-epitope antigen had a large proportion of random curls in the secondary structure,with theoretical molecular mass and isoelectric point(PI)of 13.1 ku and 7.16,which were stable and hydrophilic.It had good immunogenicity and could activate humoral and cellular immune re-sponses.The proteins prepared by ligating pET-28a(+)and pET-32a vectors with antigenic sequences were expressed as inclusion body proteins and soluble proteins,respectively.A pair of paired antibodies was obtained by animal immunization and hybridoma tech-nique,and applied to colloidal gold test strips with a sensitivity of 0.5 ng/ml,and the test strips could specifically bind two genotypes of NoV recombinant capsid proteins.Conclusion:The successful preparation and characterization of multi-epitope antigen of norovirus provides a reference for the subsequent exploration of NoV universal detection targets and the development of diagnostic raw materials.
3.A fluorescence immunochromatography method for detection of human papillomavirus type 16 E6 and L1 proteins.
Xin'er LIU ; Yinzhen ZHAO ; Nannan NIU ; Lingke LI ; Xueli DU ; Jinxiang GUO ; Yingfu ZHANG ; Jichuang WANG ; Yiqing ZHANG ; Yunlong WANG
Chinese Journal of Biotechnology 2024;40(11):4266-4276
This study aims to establish a time-resolved fluorescence immunochromatography method for simultaneous determination of human papillomavirus (HPV) type 16 E6 and L1 protein concentrations. The amount of lanthanide microsphere-labeled antibodies, the concentration of coated antibodies, and the reaction time were optimized, and then a test strip for the simultaneous determination of the protein concentrations was prepared. The performance of the detection method was evaluated based on the concordance of the results from clinical practice. The optimal conditions were 8 μg and 10 μg of HPV16 L1 and E6-labeled antibodies, respectively, 1.5 mg/mL coated antibodies, and reaction for 10 min. The detection with the established method for L1 and E6 proteins showed the linear ranges of 5-320 ng/mL and 2-64 ng/mL and the lowest limits of detection of 1.78 ng/mL and 1.09 ng/mL, respectively. There was no cross reaction with human immunodeficiency virus (HIV), treponema pallidum (TP), or HPV18 E6 and L1 proteins. The average recovery rate of the established method was between 97% and 107%. The test strip prepared in this study showed the sensitivity, specificity, and diagnostic accuracy of 97.46%, 90.57%, and 95.32%, respectively, in distinguishing patients with cervical cancer and precancerous lesions from healthy subjects, with the area under the curve (AUC) of 0.980 1 and 95% Confidence Interval (CI) of 0.956 5 to 1.000 0. The time-resolved fluorescence immunochromatography combined with the test strips prepared in this study showed high sensitivity, high accuracy, simple operation, and rapid reaction in the quantitation of HPV16 E6 and L1 proteins. It thus can be used as an auxiliary method for the diagnosis and early screening of cervical cancer and precancerous lesions and the assessment of disease course.
Oncogene Proteins, Viral/immunology*
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Humans
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Chromatography, Affinity/methods*
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Female
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Human papillomavirus 16
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Repressor Proteins/immunology*
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Capsid Proteins/immunology*
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Papillomavirus Infections/diagnosis*
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Fluorescence
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Uterine Cervical Neoplasms/virology*
4.Study on relationship between working pressure and mental health for nurses of transfusion room in paediatric department
Yinzhen YU ; Hongxia YANG ; Xin ZHAO
Chinese Journal of Practical Nursing 2012;28(25):20-21
Objective To learn the relationship between working pressure and mental health for nurse of transfusion room in pediatric department.Methods 120 nurses of transfusion room whose working years above half a year were investigated by questionnaire from January 2010 to December 2011.Results The maximal score of working pressure was working environment and resource (3.27±0.47),while the minimum was interpersonal relationship (1.80±0.68).The score of SCL-90 of the research group was higher than domestic model.More working pressures of transfusion room in pediatric department were positive correlated with SCL-90.Conclnsions In order to keep nurse sound in body and mind,the working pressure should be desolved in many ways.

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