1.Study on changes of Tfh cells and their related molecules in mouse model of premature ovarian insufficiency
Aisikaer MAIERHABA ; Yuejie ZHU ; Manli ZHANG ; Yunian ZHANG ; Hui WANG ; Xiaoyun GONG ; Jianbing DING
Chinese Journal of Immunology 2025;41(10):2313-2317
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.
2.Study on changes of Tfh cells and their related molecules in mouse model of premature ovarian insufficiency
Aisikaer MAIERHABA ; Yuejie ZHU ; Manli ZHANG ; Yunian ZHANG ; Hui WANG ; Xiaoyun GONG ; Jianbing DING
Chinese Journal of Immunology 2025;41(10):2313-2317
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.
3.Expression levels of PD-1 and PD-L1 in peripheral blood and their correlation with cytokines in patients with polycystic ovary syndrome
Rui HAN ; Xiaoyun GONG ; Yuejie ZHU ; Qifeng LI ; Erdengqieqieke YE ; Jianbing DING ; Xiaolin LA
Chinese Journal of Reproduction and Contraception 2024;44(5):480-487
Objective:To investigate the association between programmed cell death protein 1 (PD-1) and its ligand PD-L1 and cytokines in patients with polycystic ovary syndrome (PCOS).Methods:Using the GSE54248 dataset from the GEO database, differentially expressed PD1/PD-L1 pathway-related genes in PCOS were identified and subjected to GO and KEGG pathway enrichment analysis. In this case-control study, totally 105 patients with PCOS (named PCOS group) and 109 non-PCOS patients (named control group) who were treated at the Reproductive Assisted Reproduction Center of the First Affiliated Hospital of Xinjiang Medical University from January 2022 to June 2023 were recruited. The QBPlex flow cytometry high-throughput multiplex assay was utilized to assess the peripheral blood levels of PD-L1, PD-L2, PD-1, and cytokines in PCOS group and control group. Pearson's method was used for correlation analysis.Results:In PCOS group, the PD-1 level in peripheral blood [2.890 (0.020, 4.540) ng/L] was significantly lower than that of control group [3.370 (2.460, 4.360) ng/L, P=0.008], the PD-L1 level [9.820 (8.860, 10.880) ng/L] was lower than that in control group [10.410 (9.700, 11.160) ng/L, P=0.001]. There was no significant difference in the expression level of PD-L2 between the two groups ( P>0.05). From the GSE54248 dataset, 26 differentially expressed genes were identified, primarily enriched in the PD-1/PD-L1 pathway, Th1 and Th2 cell differentiation, and pathways associated with the production of cytokines involved in inflammatory responses. Compared with control group, PCOS group exhibited a significant decrease in the peripheral blood concentrations of interleukin (IL)-5, IL-9, IL-25, IL-10, growth stimulation expressed gene 2 (ST-2), and Granzyme B, and a significant increase in IL-8, IL-1RA, and tumor necrosis factor-α (TNF-α) levels, with all differences being statistically significant (all P<0.05). PD-1 exhibited positive correlations with the levels of IL-1RA, ST-2, and TNF-α ( r=0.270, P=0.005; r=0.213, P=0.029; r=0.291, P=0.003), while it exhibited negative correlations with the levels of IL-9, IL-25, and Granzyme B ( r=-0.322, P<0.001; r=-0.211, P=0.031; r=-0.369, P<0.001). PD-L1 demonstrated positive correlations with the levels of IL-9, IL-25, and Granzyme B ( r=0.254, P=0.009; r=0.330, P<0.001; r=0.340, P<0.001), and a negative correlation with IL-10 level ( r=-0.373, P=0.009). Conclusion:The expression of PD-1 and PD-L1 in the peripheral blood of PCOS patients is down-regulated, which may be associated with an imbalance in Th1/Th2 cytokines and serve as potential molecular biomarkers for the treatment of PCOS.
4.Expression levels of PD-1 and PD-L1 in peripheral blood and their correlation with cytokines in patients with polycystic ovary syndrome
Rui HAN ; Xiaoyun GONG ; Yuejie ZHU ; Qifeng LI ; Erdengqieqieke YE ; Jianbing DING ; Xiaolin LA
Chinese Journal of Reproduction and Contraception 2024;44(5):480-487
Objective:To investigate the association between programmed cell death protein 1 (PD-1) and its ligand PD-L1 and cytokines in patients with polycystic ovary syndrome (PCOS).Methods:Using the GSE54248 dataset from the GEO database, differentially expressed PD1/PD-L1 pathway-related genes in PCOS were identified and subjected to GO and KEGG pathway enrichment analysis. In this case-control study, totally 105 patients with PCOS (named PCOS group) and 109 non-PCOS patients (named control group) who were treated at the Reproductive Assisted Reproduction Center of the First Affiliated Hospital of Xinjiang Medical University from January 2022 to June 2023 were recruited. The QBPlex flow cytometry high-throughput multiplex assay was utilized to assess the peripheral blood levels of PD-L1, PD-L2, PD-1, and cytokines in PCOS group and control group. Pearson's method was used for correlation analysis.Results:In PCOS group, the PD-1 level in peripheral blood [2.890 (0.020, 4.540) ng/L] was significantly lower than that of control group [3.370 (2.460, 4.360) ng/L, P=0.008], the PD-L1 level [9.820 (8.860, 10.880) ng/L] was lower than that in control group [10.410 (9.700, 11.160) ng/L, P=0.001]. There was no significant difference in the expression level of PD-L2 between the two groups ( P>0.05). From the GSE54248 dataset, 26 differentially expressed genes were identified, primarily enriched in the PD-1/PD-L1 pathway, Th1 and Th2 cell differentiation, and pathways associated with the production of cytokines involved in inflammatory responses. Compared with control group, PCOS group exhibited a significant decrease in the peripheral blood concentrations of interleukin (IL)-5, IL-9, IL-25, IL-10, growth stimulation expressed gene 2 (ST-2), and Granzyme B, and a significant increase in IL-8, IL-1RA, and tumor necrosis factor-α (TNF-α) levels, with all differences being statistically significant (all P<0.05). PD-1 exhibited positive correlations with the levels of IL-1RA, ST-2, and TNF-α ( r=0.270, P=0.005; r=0.213, P=0.029; r=0.291, P=0.003), while it exhibited negative correlations with the levels of IL-9, IL-25, and Granzyme B ( r=-0.322, P<0.001; r=-0.211, P=0.031; r=-0.369, P<0.001). PD-L1 demonstrated positive correlations with the levels of IL-9, IL-25, and Granzyme B ( r=0.254, P=0.009; r=0.330, P<0.001; r=0.340, P<0.001), and a negative correlation with IL-10 level ( r=-0.373, P=0.009). Conclusion:The expression of PD-1 and PD-L1 in the peripheral blood of PCOS patients is down-regulated, which may be associated with an imbalance in Th1/Th2 cytokines and serve as potential molecular biomarkers for the treatment of PCOS.
5.Risk Analysis and Study of Post-marketing Adverse Events for Absorbable Sutures.
Yan WU ; Xiaoqing SUN ; Xuelei GONG ; Dong LI ; Ye ZENG ; Jianbing YIN
Chinese Journal of Medical Instrumentation 2023;47(5):571-575
Objective To investigate, analyze, and evaluate the risk data associated with the clinical use of absorbable sutures by retrieving and summarizing information from the databases of the US FDA and CNKI, as well as the adverse event reports related to absorbable sutures from January 2019 to October 2022 within Zhejiang province. The adverse event reports are obtained from both incident locations and monitoring organizations affiliated with the registrant. The aim is to identify the main risk factors associated with the clinical use of absorbable sutures. The key risk factors are potential product quality defects, product design and material selection, clinical selection and application, and postoperative recovery care including patient's self-care. Risk control strategies are further proposed to reduce or minimize the risk of adverse events caused by this product.
Humans
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Sutures/adverse effects*
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Risk Assessment
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Risk Factors
6.Human acellular nerve allograft in repair of traumatic nerve defect of upper extremity: A preliminary observation
Bo ZHANG ; Ketong GONG ; Jianbing ZHANG ; Dake ZHU
Chinese Journal of Microsurgery 2022;45(2):139-143
Objective:To observe clinical outcomes of the repair of traumatic nerve defects in the proximal upper extremities by human acellular nerve allograft(hANG).Methods:Nerve defects in the upper extremities in 8 patients were repaired by hANG from March 2017 to January 2019. The patients were 6 males and 2 females with mean age of 35.4 (21-53) years old. The nerve defects were 2 radial nerve in distal upper arm, 4 median nerve in forearm, 1 interosseous dorsal nerve and 1 ulnar nerve in forearm. All injuries were acute nerve injury. Two patients had combined injury of upper arm muscle, 4 of forearm muscle and 1 of brachial artery defect. All wound were moderate to severe contaminated. The length of nerve defects was 30-60 (mean 45) mm. The surgical procedures were fixation of fracture, repair of the muscle and discovery of the broken ends of nerve and to repair with hANG. The postoperative follow-up period ranged 18 to 40 (mean 30.6) months to observe the local response of recovery. The efficacy was evaluated by the Upper Extremity Function Evaluation Standard set up by Hand Surgery Branch of Chinese Medical Association and Grading Standard of Muscle Strength.Results:No graft rejection was observed in all cases. Primarily healing was in 5 patients. Delayed healing in 2 patients and free skin grafting was performed. Local flap transfer was performed to repair the wound in 1 patient who developed a skin necrosis 10 days after surgery. Two patients with median nerve defects had nerve function restored well. The strength of finger grip and thumb opposition muscle restored to grade IV and the sensory function had restored S 3+. The interosseous dorsal nerve in 1 patient restored well. The strength of extensor digitorum tendon had restored to grade IV. Based on the evaluation criteria for the upper extremity issued by the Hand Surgery of the Chinese Medical Association, 3 patients was rated in excellent for function recovery, 1 in fair and 4 in poor. Conclusion:After throughout debridement, hANG can be applied in the repair of traumatic nerve defect in the proximal upper extremity in an emergency surgery and it can partially restore the nerve function.
7.Cloning, expression, purification and identification of EgG1Y162-2 gene from Echinococcus granulosus
Huifang KONG ; Shangqi ZHAO ; Yanxia ZHOU ; Qiaoqiao GONG ; Yujiao LI ; Chunbao CAO ; Haimei MA ; Jianbing DING ; Xiaotao ZHOU
Chinese Journal of Endemiology 2021;40(8):635-639
Objective:To construct the pET30a-EgG1Y162-2 prokaryotic expression plasmid and induce the expression of EgG1Y162-2 protein, so as to provide a research basis for development of Echinococcus granulosus vaccine. Methods:Using Echinococcus granulosus cDNA as a template, the target gene of EgG1Y162-2 was synthesized by PCR, and after digestion with restriction enzymes EcoRⅠ and Hind Ⅲ, it was connected to the prokaryotic expression vector pET30a to construct the recombinant plasmid pET30a-EgG1Y162-2. The recombinant plasmid was transformed into competent cell BL21 (DE3) and induced by isopropyl β-D-thiogalactoside (IPTG) to express a large number of proteins. The recombinant protein was purified by affinity chromatography. The purification level was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and the expression product was identified by Western blotting. Results:The recombinant plasmid pET30a-EgG1Y162-2 was successfully constructed. After inducting expression, the bacterial supernatant and the eluate were both at a relative molecular weight of about 15 × 10 3, and the protein antigen component eluted with 200 mmol/L imidazole was relatively pure. Western blotting results showed that the purified recombinant protein EgG1Y162-2 with His tag could be recognized by His monoclonal antibody. Conclusion:The pET30a-EgG1Y162-2 prokaryotic expression plasmid of Echinococcus granulosus is successfully constructed, and the recombinant protein of EgG1Y162-2 is induced to express, laying a foundation for further study on anti- Echinococcus granulosus vaccine.
8. The value of collateral circulation classification with 4-dimensional MR angiography in predicting the short-term outcome in patients with acute ischemic stroke
Tianle WANG ; Li ZHU ; Shenchu GONG ; Jianbing YIN ; Xiaolong LI ; Haitao CHEN ; Jia LI
Chinese Journal of Radiology 2018;52(8):569-574
Objective:
To assess the collateral grade based on 4-dimensional magnetic resonance angiography (4D-MRA) in predicting short-term outcome in patients with acute ischemic stroke (AIS) .
Methods:
Forty-seven patients with unilateral MCA stroke were enrolled in this study. All patients underwent multi-modality MRI within 4.5 to 24.0 hours onset of stroke. The collateral grade was assessed through the dynamic MRI angiograms derived from perfusion raw data. The AIS patients were divided into favorable and unfavorable outcome group according to the improvement of national institutes of health stroke scale (NIHSS) score. The differences in baseline data, infarct volume, the ratio of volume of ischemic tissue on perfusion to infarct core on diffusion (rVPD) and collateral grade between the two groups were analyzed. Logistic regression analysis was used to identify variable influencing the short-term clinical outcomes. The Kappa coefficient was used to analyze the consistency of the collateral grade assessed between the two observers. Spearman rank-order correlation test was used to analyze the relationship between the collateral grade, infarct volume, hypoperfusion volume and rVPD.
Results:
The collateral grade used ASITN/SIR based on 4D-MRA was performed in 47 patients. Grade 1 in 4 patients, grade 2 in 21paitients, grade 3 in 16 patients and grade 4 in 6 patients, respectively. The collateral grade had a high consistency among the observers (Kappa=0.806,
9.Application of the interactive reading mode of PBL combined with MDT in medical imaging post-graduate clinical teaching
Wu CAI ; Jianping GONG ; Zhen JIANG ; Jianbing ZHU ; Guangqiang CHEN ; Fang QIAO ; Xin DOU ; Jian HUAN ; Wei ZHANG ; Junkang SHEN
Chinese Journal of Medical Education Research 2016;15(9):947-950
Medical imaging is an interdisciplinary subject closely related to clinical and pathological subject. Its clinical reading skills' training has become the focus of postgraduate teaching. In the process of clinical teaching, the interactive reading mode of problem-based learning (PBL) combined with multi-disci-plinary team (MDT) was introduced into clinical reading meeting. The tutors chose the reading cases proved by pathology; designed in-depth issues step by step for execution of PBL teaching; guided postgraduates to delineate imaging signs and propose the diagnostic results, evidences and differential diagnoses according to the step from localizing to qualitative and then to pathological diagnosis;then guided postgraduates to attend in-depth case analysis of MDT and analyze the correlation or inconsistency between the imaging diagnosis and clinical and pathological diagnosis; exercise document retrieval and verbalization, multimedia design, and writing level of the records of the reading cases and papers. The interactive reading mode of PBL com-bined with MDT has achieved significant effects, which is worthy of further exploration and promotion.
10.The effect of epicardial fat volume in patients with metabolic syndrome on insulin resistance and coronary heart disease
Qiang WANG ; Tao HANG ; Yun XU ; Xunming CHENG ; Qigao ZHANG ; Jianbing GONG ; Jing WANG
Journal of Medical Postgraduates 2014;(4):390-393
Objective Epicardial fat volume ( EFV) was a risk factor for coronary heart disease ( CHD) , but there is little research regarding the relationship of EFV with insulin resistance ( IR) and CHD in patients with metabolic syndrome ( MS) .The aim of the article was to explore the effect of EFV in patients with MS on CHD and IR . Methods Patients with MS treated by percutane-ous coronary angiography ( CAG) in our hospital from February 2013 to August 2013 were recruited in this study .The data of height , weight, waist circumference(WC) and hip circumference(HP) were recorded.EFV were measured by MSCT.Fasting blood samples were collected for blood biochemical test . Results EFV in patients with MS was in positive relation with IR index (IRI)(r=0.335, P<0.001) and CHD (r=0.321, P<0.05), and the correlation still remained when the influences of WC and body mass index (BMI) were excluded.Logistic regression analysis showed that EFV was an independent risk factor for CHD (P<0.05), while linear regression analysis indicated EFV , BMI and LDL-C were the risk factors for IRI .ROC curves analysis proved EFV and BMI had diag-nosis value for IRI, and the areas under curve of EFV were 0.755 and 0.679 (P<0.05) respectively. Conclusion EFV is an in-dependent risk factor for CHD and IRI in patients with MS , and EFV has an advantage over BMI in the diagnosis value of IRI .

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