To prepare monoclonal antibody to the N protein of porcine reproductive and respiratory syndrome virus(PRRSV),BALB/c mice were immunized with the purified N protein of the PRRSV-2 strain expressed by prokaryotic expression system.Mouse splenocytes were fused with myeloma cells using hybridoma technique,hybridoma cells were identified by indirect ELISA method and indirect immunofluorescence assay(IFA),and positive hybridoma cells were screened for subclones using the limited dilution method.The results showed that a monoclonal antibody cell line 4A7 was successfully obtained,and the results of Western blot and IFA indicated that the monoclonal antibody could accurately recognize the N proteins of type 1 and type 2 PRRSV.Mean-while,the N protein gene was truncated and expressed by prokaryotic expression system,and the amino acid sequence of the B-cell antigenic epitope recognized by 4A7 was screened as 51EKPHF55 using Western blot.Comparison of epitope amino acids in the N protein gene sequences of different strains revealed that the antigenic epitope 51EKPHF55 recognized by the monoclonal antibody 4A7 has no amino acid difference in the sequences of three subtypes among the PRRSV-1 strains and nine lineages among the PRRSV-2 strains,indicating a high degree of conservation.The results of the study provide a foundation the development of PRRSV diagnostic kits and novel vaccines.

Objective:To understand the incidence of adverse pregnancy outcome in HIV-infected pregnant women and influencing factors in China and provide reference for the improvement of the health status of HIV-infected pregnant women and their newborns.Methods:Based on a mother-child cohort of HIV-infected pregnant women and children (PMTCT-MC-2005) established in Guangxi Zhuang Autonomous Region, Yunnan Province and Xinjiang Uygur Autonomous Region, this study enrolled pregnant women with or without HIV infection as study subjects from January 2017 to June 2023, a total of 1 646 pregnant women (558 HIV-infected and 1 088 HIV-uninfected) were included, and 34 cases with missing data were excluded. The χ2 test was used to analyze the difference in the incidence adverse pregnancy outcome between two groups, and used logistic regression model to identify the influencing factors of adverse pregnancy outcome in HIV-infected pregnant women. Results:A total of 1 612 pregnant women were included in the study, in whom 541 were infected with HIV and 1 071 were not infected with HIV. The incidence of adverse pregnancy outcome was 18.8% (303/1 612), the incidence of adverse pregnancy outcome was 33.1% (179/541) in the HIV-infected pregnant women and 11.6% (124/1 071) in the pregnant women without HIV infection. The results of multivariable logistic regression analysis showed that the influencing factors of adverse pregnancy outcome were age <35 years at delivery (a OR=0.64, 95% CI: 0.43-0.95) compared with the age ≥35 years and the duration of antiviral treatment over 10 years (a OR=0.43, 95% CI: 0.23-0.79) compared with less than one year. Conclusions:The incidence of adverse pregnancy outcome in HIV-infected pregnant women was high in some regions of China during 2017-2023. It is necessary for HIV-infected women to get pregnancy at appropriate time based on antiretroviral treatment effect and strengthen self-care to reduce the incidence of adverse pregnancy outcome.

Objective:To understand the incidence of adverse pregnancy outcome in HIV-infected pregnant women and influencing factors in China and provide reference for the improvement of the health status of HIV-infected pregnant women and their newborns.Methods:Based on a mother-child cohort of HIV-infected pregnant women and children (PMTCT-MC-2005) established in Guangxi Zhuang Autonomous Region, Yunnan Province and Xinjiang Uygur Autonomous Region, this study enrolled pregnant women with or without HIV infection as study subjects from January 2017 to June 2023, a total of 1 646 pregnant women (558 HIV-infected and 1 088 HIV-uninfected) were included, and 34 cases with missing data were excluded. The χ2 test was used to analyze the difference in the incidence adverse pregnancy outcome between two groups, and used logistic regression model to identify the influencing factors of adverse pregnancy outcome in HIV-infected pregnant women. Results:A total of 1 612 pregnant women were included in the study, in whom 541 were infected with HIV and 1 071 were not infected with HIV. The incidence of adverse pregnancy outcome was 18.8% (303/1 612), the incidence of adverse pregnancy outcome was 33.1% (179/541) in the HIV-infected pregnant women and 11.6% (124/1 071) in the pregnant women without HIV infection. The results of multivariable logistic regression analysis showed that the influencing factors of adverse pregnancy outcome were age <35 years at delivery (a OR=0.64, 95% CI: 0.43-0.95) compared with the age ≥35 years and the duration of antiviral treatment over 10 years (a OR=0.43, 95% CI: 0.23-0.79) compared with less than one year. Conclusions:The incidence of adverse pregnancy outcome in HIV-infected pregnant women was high in some regions of China during 2017-2023. It is necessary for HIV-infected women to get pregnancy at appropriate time based on antiretroviral treatment effect and strengthen self-care to reduce the incidence of adverse pregnancy outcome.

Objective:To analyze the application effect of the patient navigation model in the continuity of care for elderly diabetic patients.Methods:A convenience sampling method was used to select 163 elderly diabetic patients admitted to the First Affiliated Hospital of Zhengzhou University from June 2022 to December 2023. Patients were randomly divided into a control group ( n=81) and an observation group ( n=82) using a random number table. The control group received routine continuity of care, while the observation group received continuity of care based on the patient navigation model. The Diabetes Self-Management Behavior Scale (SDSCA) and the Chronic Disease Resources Survey were used to assess the patients. Self-management behaviors, chronic disease resource utilization, and blood glucose changes were compared between the two groups before and three months after the intervention. Results:After the intervention, the SDSCA dimension scores in the observation group were significantly higher than those in the control group ( P<0.01). Additionally, the chronic disease resource survey dimension scores in the observation group were significantly higher than those in the control group ( P<0.01). Moreover, the fasting blood glucose, 2-hour postprandial blood glucose, and HbA1c levels in the observation group were significantly lower than those in the control group ( P<0.01) . Conclusions:The application of the patient navigation model in the continuity of care for elderly diabetic patients helps patients fully access multi-dimensional social resources, improves self-management capabilities, and is beneficial for improving blood glucose control.

To prepare monoclonal antibody to the N protein of porcine reproductive and respiratory syndrome virus(PRRSV),BALB/c mice were immunized with the purified N protein of the PRRSV-2 strain expressed by prokaryotic expression system.Mouse splenocytes were fused with myeloma cells using hybridoma technique,hybridoma cells were identified by indirect ELISA method and indirect immunofluorescence assay(IFA),and positive hybridoma cells were screened for subclones using the limited dilution method.The results showed that a monoclonal antibody cell line 4A7 was successfully obtained,and the results of Western blot and IFA indicated that the monoclonal antibody could accurately recognize the N proteins of type 1 and type 2 PRRSV.Mean-while,the N protein gene was truncated and expressed by prokaryotic expression system,and the amino acid sequence of the B-cell antigenic epitope recognized by 4A7 was screened as 51EKPHF55 using Western blot.Comparison of epitope amino acids in the N protein gene sequences of different strains revealed that the antigenic epitope 51EKPHF55 recognized by the monoclonal antibody 4A7 has no amino acid difference in the sequences of three subtypes among the PRRSV-1 strains and nine lineages among the PRRSV-2 strains,indicating a high degree of conservation.The results of the study provide a foundation the development of PRRSV diagnostic kits and novel vaccines.

Objective:To analyze the application effect of the patient navigation model in the continuity of care for elderly diabetic patients.Methods:A convenience sampling method was used to select 163 elderly diabetic patients admitted to the First Affiliated Hospital of Zhengzhou University from June 2022 to December 2023. Patients were randomly divided into a control group ( n=81) and an observation group ( n=82) using a random number table. The control group received routine continuity of care, while the observation group received continuity of care based on the patient navigation model. The Diabetes Self-Management Behavior Scale (SDSCA) and the Chronic Disease Resources Survey were used to assess the patients. Self-management behaviors, chronic disease resource utilization, and blood glucose changes were compared between the two groups before and three months after the intervention. Results:After the intervention, the SDSCA dimension scores in the observation group were significantly higher than those in the control group ( P<0.01). Additionally, the chronic disease resource survey dimension scores in the observation group were significantly higher than those in the control group ( P<0.01). Moreover, the fasting blood glucose, 2-hour postprandial blood glucose, and HbA1c levels in the observation group were significantly lower than those in the control group ( P<0.01) . Conclusions:The application of the patient navigation model in the continuity of care for elderly diabetic patients helps patients fully access multi-dimensional social resources, improves self-management capabilities, and is beneficial for improving blood glucose control.

Objective To analyze the effect of microRNA-22-3p (miR-22-3p) of extracellular vesicles derived from bone marrow mesenchymal stem cells on premature ovarian failure. Methods Follicular fluids were provided by premature ovarian failure patients with in vitro fertilization or the second-generation in vitro fertilization treatment and normal volunteers with the same treatment for infertility due to male factors; the exosomes derived from bone marrow mesenchymal stem cells were isolated by differential centrifugation; the morphology, particle size and marker proteins of isolated exosomes were analyzed by transmission electron microscopy, NanoSight LM10 analyzer and Western blotting. Granulosa cells were treated with cyclophosphamide (CTX), exosomes, miR-22-3 mimics and corresponding controls, and were divided into CTX group, control group, exosome incubation group, PBS treatment group, CTX+miR-22-3p group, CTX+miR-NC group, CTX+Exosomes group, and CTX+PBS group; gene expression was detected by Western blotting and quantitative real-time polymerase chain reaction (qRT-PCR). The 3-(4, 5-Dimethylthazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) reagent and flow cytometry were used to detect cell viability and apoptosis. Results The extracted exosomes had typical goblet vesicles, the exosome marker proteins C cluster of differentiation 63 (CD63) and C cluster of differentiation 9 (CD9) were expressed in the extracted exosomes, and exosome particle size was 80 to 150 nm; miR-22-3p was significantly lowly expressed in patients with premature ovarian failure and ovarian granulosa cells induced by CTX (P < 0.05), but was significantly highly expressed in ovarian granulosa cells incubated with exosomes derived from bone marrow mesenchymal stem cells (P < 0.05); the activity of granulosa cells in the CTX group was significantly lower than that in the control group, but was significantly higher in the CTX+miR-22-3p group or CTX+Exosomes group than that in the control group (P < 0.05); the apoptosis rate of granulosa cells in the CTX group was significantly higher than that in the control group, but was significantly lower in the CTX+miR-22-3p or CTX+Exosomes group than that in the control group (P < 0.05). Conclusion The miR-22-3p of extracellular vesicles derived from bone marrow mesenchymal stem cells can inhibit ovarian granulosa cell injury induced by CTX.

Objective:To explore the etiology of primary amenorrhea,analyze its clinical features,and improve the level of clinical diagnosis and treatment.Methods:A retrospective analysis was conducted on the clinical data of 75 patients with primary amenorrhea who attended the gynecological endocrine clinic of Nanjing Women and Children's Healthcare Hospital from January 2020 to September 2022,and the etiology of their diagnosis was ana-lyzed.Results:aChromosomal karyotype analysis showed 14 cases(18.67％)of abnormal karyotypes,including 2 cases of autosomal abnormalities,7 cases of sex chromosome abnormalities,and 5 cases of sexual differentia-tion abnormalities.②According to reproductive hormones,there were 15 cases(20.00％)of hypogonadotropic a-menorrhea and 18 cases(24.00％)of hypergonadotropic amenorrhea.③By the location of the cause,utero-inferi-or genital tract amenorrhea was the most common in 23 cases(30.67％),including 20 cases of mayer-rokitan-sky-küster-hauser syndrome(MRKH syndrome)and 3 cases of androgen insensitivity syndrome(AIS).It was followed by ovarian amenorrhea in 18 cases(24.00％)including 6 cases of Turners syndrome,10 cases of 46,XX simple gonadal hypoplasia and 2 cases of 46,XY gonadal hypoplasia(Swyer syndrome);hypothalamic-pituitary amenorrhea in 17 cases(22.67％),including idiopathic hypogonadotropic hypogonadism(IHH)in 9 cases,Kall-mann s syndrome in 4 cases,and hyperprolactinemia in 4 cases;and also amenorrhea with hyperandrogenic mani-festations in 17 cases(22.67％),with a higher number of suspicious polycystic ovary syndrome in 16 cases.Con-clusions:Primary amenorrhea has a variety of etiologies and clinical features,and requires examination of sex hormones and pelvic ultrasound from the lower genital tract up to the central nervous system to find the site of the etiology and to optimize the diagnostic process;in the case of hypogonadotropic amenorrhea,chromosomal test is necessary.

Objective:To explore the etiology of primary amenorrhea,analyze its clinical features,and improve the level of clinical diagnosis and treatment.Methods:A retrospective analysis was conducted on the clinical data of 75 patients with primary amenorrhea who attended the gynecological endocrine clinic of Nanjing Women and Children's Healthcare Hospital from January 2020 to September 2022,and the etiology of their diagnosis was ana-lyzed.Results:aChromosomal karyotype analysis showed 14 cases(18.67％)of abnormal karyotypes,including 2 cases of autosomal abnormalities,7 cases of sex chromosome abnormalities,and 5 cases of sexual differentia-tion abnormalities.②According to reproductive hormones,there were 15 cases(20.00％)of hypogonadotropic a-menorrhea and 18 cases(24.00％)of hypergonadotropic amenorrhea.③By the location of the cause,utero-inferi-or genital tract amenorrhea was the most common in 23 cases(30.67％),including 20 cases of mayer-rokitan-sky-küster-hauser syndrome(MRKH syndrome)and 3 cases of androgen insensitivity syndrome(AIS).It was followed by ovarian amenorrhea in 18 cases(24.00％)including 6 cases of Turners syndrome,10 cases of 46,XX simple gonadal hypoplasia and 2 cases of 46,XY gonadal hypoplasia(Swyer syndrome);hypothalamic-pituitary amenorrhea in 17 cases(22.67％),including idiopathic hypogonadotropic hypogonadism(IHH)in 9 cases,Kall-mann s syndrome in 4 cases,and hyperprolactinemia in 4 cases;and also amenorrhea with hyperandrogenic mani-festations in 17 cases(22.67％),with a higher number of suspicious polycystic ovary syndrome in 16 cases.Con-clusions:Primary amenorrhea has a variety of etiologies and clinical features,and requires examination of sex hormones and pelvic ultrasound from the lower genital tract up to the central nervous system to find the site of the etiology and to optimize the diagnostic process;in the case of hypogonadotropic amenorrhea,chromosomal test is necessary.

Objective:To explore the etiology of primary amenorrhea,analyze its clinical features,and improve the level of clinical diagnosis and treatment.Methods:A retrospective analysis was conducted on the clinical data of 75 patients with primary amenorrhea who attended the gynecological endocrine clinic of Nanjing Women and Children's Healthcare Hospital from January 2020 to September 2022,and the etiology of their diagnosis was ana-lyzed.Results:aChromosomal karyotype analysis showed 14 cases(18.67％)of abnormal karyotypes,including 2 cases of autosomal abnormalities,7 cases of sex chromosome abnormalities,and 5 cases of sexual differentia-tion abnormalities.②According to reproductive hormones,there were 15 cases(20.00％)of hypogonadotropic a-menorrhea and 18 cases(24.00％)of hypergonadotropic amenorrhea.③By the location of the cause,utero-inferi-or genital tract amenorrhea was the most common in 23 cases(30.67％),including 20 cases of mayer-rokitan-sky-küster-hauser syndrome(MRKH syndrome)and 3 cases of androgen insensitivity syndrome(AIS).It was followed by ovarian amenorrhea in 18 cases(24.00％)including 6 cases of Turners syndrome,10 cases of 46,XX simple gonadal hypoplasia and 2 cases of 46,XY gonadal hypoplasia(Swyer syndrome);hypothalamic-pituitary amenorrhea in 17 cases(22.67％),including idiopathic hypogonadotropic hypogonadism(IHH)in 9 cases,Kall-mann s syndrome in 4 cases,and hyperprolactinemia in 4 cases;and also amenorrhea with hyperandrogenic mani-festations in 17 cases(22.67％),with a higher number of suspicious polycystic ovary syndrome in 16 cases.Con-clusions:Primary amenorrhea has a variety of etiologies and clinical features,and requires examination of sex hormones and pelvic ultrasound from the lower genital tract up to the central nervous system to find the site of the etiology and to optimize the diagnostic process;in the case of hypogonadotropic amenorrhea,chromosomal test is necessary.