Objective:Haracterization of follicular helper T cells(Tfh)cells and related molecules early onset in premature ovarian insufficiency(POI)based on a mouse model of POI.Methods:After acclimatization feeding of 30 BALB/c mice were selected,mice were subjected to vaginal exfoliative cytology from 08:00 onwards every day for 10 consecutive days under light microscope obser-vation,20 healthy female mice with regular estrous cycle were screened out and randomly divided into model(POI)group and healthy control(HC)group according to random number table method,different chemical interventions were given and processed and tissue sampling was done on the 21st day.Every morning from the beginning to the end of the modeling period,the mice were subjected to cell smears of vaginal secretions,which were stained with Giemsa's stain and then observed under a light microscope.After the mice were sacrificed,bilateral ovarian tissues were taken and ovarian paraffin sections were made,and the tissues were stained with HE to observe pathological changes in ovaries of mice.Immunohistochemistry was used to detect expressions of ovarian Tfh-related molecules CXCR5 and BCL-6;flow assay was used to detect frequency of CD4+CXCR5+Tfh cells and CD4+ICOS+Tfh cells in spleens of mice;ELISA was used to detect levels of IL-6 and IL-21 in splenic tissues of mice;qRT-PCR was used to detect mRNA levels of CXCR5 and BCL-6 in mice;correlation analysis was performed to correlate frequencies of CD4+CXCR5+Tfh and CD4+ICOS+Tfh cells with levels of IL-6 and IL-21 in spleens of mice in both groups.Results:Pathological changes in ovarian tissue of mice in POI group:primordial folli-cles and growing follicles decreased,while atretic follicles increased.BCL-6 and CXCR5 were highly expressed in ovarian tissue of mice in POI group.Frequency of CD4+CXCR5+Tfh and CD4+ICOS+Tfh cells in spleen of POI mice was higher than that in HC group.Levels of IL-6 and IL-21 in spleens of mice in POI group were significantly higher than those in HC group,and were positively correlated with the frequency of CD4+CXCR5+Tfh and CD4+ICOS+Tfh cells in spleen of mice.Conclusion:In POI mouse model,Tfh cells are highly expressed and the level of Tfh-related molecules changes,which provides basic reference for studying pathogenesis of POI.

Objective:Haracterization of follicular helper T cells(Tfh)cells and related molecules early onset in premature ovarian insufficiency(POI)based on a mouse model of POI.Methods:After acclimatization feeding of 30 BALB/c mice were selected,mice were subjected to vaginal exfoliative cytology from 08:00 onwards every day for 10 consecutive days under light microscope obser-vation,20 healthy female mice with regular estrous cycle were screened out and randomly divided into model(POI)group and healthy control(HC)group according to random number table method,different chemical interventions were given and processed and tissue sampling was done on the 21st day.Every morning from the beginning to the end of the modeling period,the mice were subjected to cell smears of vaginal secretions,which were stained with Giemsa's stain and then observed under a light microscope.After the mice were sacrificed,bilateral ovarian tissues were taken and ovarian paraffin sections were made,and the tissues were stained with HE to observe pathological changes in ovaries of mice.Immunohistochemistry was used to detect expressions of ovarian Tfh-related molecules CXCR5 and BCL-6;flow assay was used to detect frequency of CD4+CXCR5+Tfh cells and CD4+ICOS+Tfh cells in spleens of mice;ELISA was used to detect levels of IL-6 and IL-21 in splenic tissues of mice;qRT-PCR was used to detect mRNA levels of CXCR5 and BCL-6 in mice;correlation analysis was performed to correlate frequencies of CD4+CXCR5+Tfh and CD4+ICOS+Tfh cells with levels of IL-6 and IL-21 in spleens of mice in both groups.Results:Pathological changes in ovarian tissue of mice in POI group:primordial folli-cles and growing follicles decreased,while atretic follicles increased.BCL-6 and CXCR5 were highly expressed in ovarian tissue of mice in POI group.Frequency of CD4+CXCR5+Tfh and CD4+ICOS+Tfh cells in spleen of POI mice was higher than that in HC group.Levels of IL-6 and IL-21 in spleens of mice in POI group were significantly higher than those in HC group,and were positively correlated with the frequency of CD4+CXCR5+Tfh and CD4+ICOS+Tfh cells in spleen of mice.Conclusion:In POI mouse model,Tfh cells are highly expressed and the level of Tfh-related molecules changes,which provides basic reference for studying pathogenesis of POI.

Abstract: Objective To investigate the expression of T cell immunoglobulin and mucin domain-containing protein 3 (Tim-3) on the surface of T cells in patients with brucellosis (Bm), as well as the expression of interleukin-10 (IL-10) and transforming growth factor beta (TGF-β) in serum, and to analyze the differential expression of these indicators in patients with acute and chronic brucellosis, in order to provide new approaches for the differential diagnosis of acute and chronic brucellosis. Methods A total of 56 patients diagnosed with brucellosis at the First Affiliated Hospital of Xinjiang Medical University from April 2023 to September 2023 were selected, including 31 patients in the acute phase and 25 patients in the chronic phase. Additionally, 35 healthy individuals underwent routine physical examinations within the same period served as healthy controls. Flow cytometry was used to detect and compare Tim-3 levels on the CD4+ T cells' surface among the groups. Levels of serum IL-10 and TGF-β were measured and compared using CBA and ELISA, respectively, and the relationship of these factors with the staging of brucellosis patients was analyzed. Results The proportions of Tim-3+CD3+CD4+T cells in the control group, acute group, and chronic group were (2.56±1.25)%, (5.14±1.98)%, and (13.66±2.66)%, respectively. The Tim-3 levels in the patients with brucellosis were higher than those in the healthy control group, with the chronic group showing even higher levels, and these differences were statistically significant (P<0.05). The levels of IL-10 and TGF in the patient group were higher than those in the healthy control group, with the chronic group exhibiting significantly higher levels of IL-10 and TGF-β than the acute group, also presenting statistically significant differences (P<0.05). The areas under the ROC curve for predicting chronic brucellosis with Tim-3, IL-10, and TGF-β scores were 0.876, 0.865, and 0.663, respectively. Conclusions There are certain differences in the expression of Tim-3, serum IL-10, and TGF-β among patients with brucellosis, with high expression indicating a potential transition to the chronic phase of the disease. Tim-3 has shown the best diagnostic performance. Therefore, as a diagnostic indicator, Tim-3 may provide new ideas and strategies for the treatment and differential diagnosis of brucellosis.