OBJECTIVETo set up a novel, simple, sensitive, specific, repeatable and rapid assay for diagnosis of influenza.

METHODSMonolayers of MDCK cells were inoculated with the specimens for amplifying viral yield, the feature of receptors on cell surface was changed by treatment of neuraminidases of influenza A and B viruses. Afterward, based on the lectin binds to receptors on cell surface with strict specificity,the phenomenon of red blood cell aggregation was observed under the conventional microscope. Finally, the tested results could be determined by the extent of red blood cell aggregation.

RESULTSThere was a complete (%) consistency rate (100%) for viral isolation between new and routine tests. In general, the results were detected with new assay within 20 h. The sensitivity of new assay was over 100-10,000 times higher than that of routine method. Meanwhile, the novel test could not only be used for rapid diagnosis in the clinic, but also be used for influenza surveillance. The best concentration of red blood cells was 1 in the detection assay. The testing result was not effected by red blood cells taken from either different red blood cell type of human or different individual of guinea pigs.

CONCLUSIONSThe novel method has several advantages: simple, high sensitivity and specificity, accurate and suitable for multiple purposes.

Animals ; Cells, Cultured ; Erythrocyte Aggregation ; drug effects ; Guinea Pigs ; Humans ; Influenza A virus ; enzymology ; isolation & purification ; Influenza B virus ; enzymology ; isolation & purification ; Influenza, Human ; diagnosis ; virology ; Neuraminidase ; analysis ; Reagent Kits, Diagnostic ; Sensitivity and Specificity

BACKGROUNDConstructing replication defective recombinant adenovirus vector expressing the group specific antigen VP6 of human rotavirus and studying the immune responses induced in vivo.

METHODSThe cDNA of full length VP6 was inserted into the adenovirus vector pShuttle-CMV, and recombinant adenovirus genome DNA was obtained through homological recombination in E.coli,then the recombinant adenovirus was gained after transfecting 293 cell line with the genome DNA. Gene integration of VP6 in resultant adenovirus was confirmed by PCR and Southern blot, respectively gene expression was confirmed in 293 cells by Western blot. BALB/c mice were immunized intranasally(inl)and orally(ora), respectively, to test the immunization effects of the adenovirus.

RESULTSRecombinant adenovirus named rvAd-VP6 was obtained. The cDNA of VP6 was integrated in the adenovirus and was able to be expressed in 293 cells stably. The systemic immune responses to rotavirus VP6 could be induced effectively in both oral and intranasal group, the titer of serum IgG antibody in the two group of mice were 1?1 000 and 1?10 000-1?100 000, respectively. In addition to IgG, the serum IgA specific to VP6 could also be detected at a titer of 1?10-1?100. Secretory IgA(sIgA) was detected in both lung lavage fluid and intestinal homogenate when administered intranasally to BALB/c mice, whereas only found in intestinal homogenate in the oral group. The results indicated that the immunization efficacy of intranasal inoculation was superior to that of oral inoculation.

CONCLUSIONSThe recombinant adenovirus vector expressing human rotavirus VP6 was successfully constructed, its ability to induce immune responses has laid a solid foundation for the development of rotavirus genetically engineering vaccine against rotavirus infection.

Adenoviridae ; genetics ; Animals ; Antibodies, Viral ; blood ; Antigens, Viral ; Capsid Proteins ; biosynthesis ; immunology ; Genetic Vectors ; Mice ; Mice, Inbred BALB C ; Recombination, Genetic ; Rotavirus ; immunology

OBJECTIVES: This study examined the associations between adverse childhood experiences (ACEs) and diabetes within a social-ecological framework, incorporating personal and environmental unfavorable conditions during childhood from family, school, and community contexts. METHODS: Data were obtained from the China Health and Retirement Longitudinal Study (2014 life history survey and 2015 survey), including 9,179 participants aged ≥45 years. ACEs were collected through self-report questionnaires, and participants were categorized based on the number of distinct ACEs experienced (0, 1, 2, 3, or ≥4 ACEs). Diabetes was defined by biomarkers, self-reported diagnosis, and treatment status. Logistic regression was conducted to explore the associations between ACEs and diabetes. Subgroup analyses were conducted by gender, age, and obesity status. RESULTS: Compared with participants without ACEs, those exposed to any ACE (odds ratio [OR], 1.19; 95% confidence interval [CI], 1.01 to 1.40), 3 ACEs (OR, 1.32; 95% CI, 1.07 to 1.62) and ≥4 ACEs (OR, 1.29; 95% CI, 1.07 to 1.56) had an increased risk of diabetes. For each additional ACE, the risk of diabetes increased by about 5%. Regarding the source of ACEs, those originating from the family (OR, 1.23; 95% CI, 1.08 to 1.41) were associated with diabetes. In terms of specific ACE types, family members with substance abuse (OR, 1.23; 95% CI, 1.01 to 1.52), emotional abuse (OR, 1.28; 95% CI, 1.12 to 1.46), and poor parental relationship (OR, 1.25; 95% CI, 1.09 to 1.43) were associated with diabetes. CONCLUSIONS ACEs, particularly those originating from the family, were associated with diabetes. Interventions aimed at preventing and mitigating ACEs are essential for the early prevention of diabetes.

Objective To draw a Levey-Jennings quality control chart using influenza virus RNA as internal quality control substance.Method Recent popular representative strains of influenza virus were selected and extracted RNA.Make series 10 folds dilution and take different dilution to do quantitative RTPCR.Three parallel wells of each dilution were done.High Ct value and low Ct value corresponding dilution of RNA were selected and detected continuously twenty times.Mean Ct value (x) and standard deviation (s) were calculated.Levey-Jennings quality control chart were drew by using Excel software.Result Levey-Jennings quality control charts of three influenza virus subtype were drew and each internal quality control substance was in control.Conclusion RNA quality substance of each subtype has good stability and preparation simple which is convenient to draw quality control chart and suitable as internal quality control substance for PCR laboratory.