Steroid-induced avascular necrosis of femoral head（SANFH） is a bone and joint disease caused by prolonged and excessive steroid use. Its typical pathological features involve progressive circulatory disorders in the blood supply system of femoral head， leading to osteocyte apoptosis and bone tissue necrosis. As the disease progresses， it ultimately results in structural collapse and necrotic lesions of the femoral head， severely affecting patients' limb function and quality of life. Glucocorticoids mediate pathological damage through dual mechanisms， on the one hand， they disrupt the dynamic equilibrium between bone formation and resorption by suppressing osteoblast differentiation activity and activating osteoclastogenesis， on the other hand， they induce lipid metabolism disorders， inhibit angiogenesis， and impair endothelial cell function， thereby triggering microcirculatory disorders. Epimedii Folium and its active components exhibit multidimensional regulatory effects in SANFH prevention and treatment. Literature review reveals that it is rich in multiple active ingredients， primarily including total flavonoids of Epimedii Folium， icaritin， icariin， kaempferol， icariside Ⅱ， etc. These compounds exert multiple pharmacological effects（regulating bone metabolic homeostasis， modulating angiogenesis， correcting lipid metabolism disorders， and controlling cellular autophagy processes） through multiple signaling pathways， including Wnt/β-catenin， transforming growth factor（TGF）-β/bone morphogenetic protein（BMP）/Smad， mitogen-activated protein kinase（MAPK）， phosphoinositide 3-kinase/protein kinase B（PI3K/Akt）， osteoprotegerin/receptor activator of nuclear transcription factor-κB ligand/receptor activator of nuclear transcription factor-κB（OPG/RANKL/RANK）， etc. Based on existing research findings， this paper systematically elucidates the intervention mechanisms of active components in Epimedii Folium on key pathological processes of SANFH through the above pathways. It also deeply analyzes their regulatory roles in key nodes of different signaling pathways， aiming to provide valuable references for future clinical treatment and experimental research.

Objective To investigate the protective effect and mechanism of Polygonatum sibiricum polysaccharides(PSP)on diabetic cardiomyopathy(DCM).Methods Forty SPF-grade male Sprague-Dawley rats were divided randomly into Control,Model,PSP,and metformin groups.After 4 weeks of feeding a high-fat diet,streptozotocin was injected intraperitoneally to establish a rat model of diabetes mellitus.The drug was administered by gavage for 12 weeks,and body mass and blood glucose were recorded every 2 weeks.Cardiac function was detected by non-invasive echocardiography at week 16.Myocardial histopathological changes and the degree of myocardial fibrosis were assessed by hematoxylin and eosin and Masson staining.Serum interleukin(IL)-6,IL-1β,IL-18,tumor necrosis factor-α(TNF-α),triglycerides,total cholesterol,low-density lipoprotein,and high-density lipoprotein were detected by enzyme-linked immunosorbent assay.Expression levels of the fibrosis-related proteins transforming growth factor(TGF)-β1,Smad2,Collagen-Ⅰ,Collagen-Ⅲ,and the pyroptosis-related proteins NOD-like receptor thermal protein domain associated protein 3(NLRP3),apoptosis-associated speck-like protein containing a caspase recruitment domain(ASC),and Caspase-1 were detected in rat myocardial tissues by Western blot.Cellular experiments were performed by exposing H9c2 cells to high glucose(40 mmol/L)to mimic the in vitro DCM model,cell viability was detected by Cell Counting Kit-8 assay,and the apoptotic cell ratio was detected by flow cytometry.Results Rats in the treatment group had significantly lower blood glucose,lipid,and serum inflammatory factor levels compared with the model group(P＜0.05),significantly higher ejection fraction and fractional shortening values(P＜0.05),and improved cardiac function.Myocardial fibers were better aligned and collagen fiber accumulation was reduced,and myocardial tissue levels of NLRP3,ASC,Caspase-1,Collagen-Ⅰ,Collagen-Ⅲ,TGF-β1,and Smad2 were significantly reduced(P＜0.05).In the cellular assay,PSP increased the viability and decreased the proportion of apoptotic cells in high glucose-induced H9c2 cardiomyocytes.Conclusions PSP can improve glucose-lipid metabolism,protect cardiac function,and delay the occurrence of myocardial fibrosis in diabetic rats,and can also improve the viability of cardiomyocytes.Its mechanism of action may be related to the inhibition of cellular pyroptosis and delayed occurrence of ventricular remodeling.

Objective To investigate the protective effect and mechanism of Polygonatum sibiricum polysaccharides(PSP)on diabetic cardiomyopathy(DCM).Methods Forty SPF-grade male Sprague-Dawley rats were divided randomly into Control,Model,PSP,and metformin groups.After 4 weeks of feeding a high-fat diet,streptozotocin was injected intraperitoneally to establish a rat model of diabetes mellitus.The drug was administered by gavage for 12 weeks,and body mass and blood glucose were recorded every 2 weeks.Cardiac function was detected by non-invasive echocardiography at week 16.Myocardial histopathological changes and the degree of myocardial fibrosis were assessed by hematoxylin and eosin and Masson staining.Serum interleukin(IL)-6,IL-1β,IL-18,tumor necrosis factor-α(TNF-α),triglycerides,total cholesterol,low-density lipoprotein,and high-density lipoprotein were detected by enzyme-linked immunosorbent assay.Expression levels of the fibrosis-related proteins transforming growth factor(TGF)-β1,Smad2,Collagen-Ⅰ,Collagen-Ⅲ,and the pyroptosis-related proteins NOD-like receptor thermal protein domain associated protein 3(NLRP3),apoptosis-associated speck-like protein containing a caspase recruitment domain(ASC),and Caspase-1 were detected in rat myocardial tissues by Western blot.Cellular experiments were performed by exposing H9c2 cells to high glucose(40 mmol/L)to mimic the in vitro DCM model,cell viability was detected by Cell Counting Kit-8 assay,and the apoptotic cell ratio was detected by flow cytometry.Results Rats in the treatment group had significantly lower blood glucose,lipid,and serum inflammatory factor levels compared with the model group(P＜0.05),significantly higher ejection fraction and fractional shortening values(P＜0.05),and improved cardiac function.Myocardial fibers were better aligned and collagen fiber accumulation was reduced,and myocardial tissue levels of NLRP3,ASC,Caspase-1,Collagen-Ⅰ,Collagen-Ⅲ,TGF-β1,and Smad2 were significantly reduced(P＜0.05).In the cellular assay,PSP increased the viability and decreased the proportion of apoptotic cells in high glucose-induced H9c2 cardiomyocytes.Conclusions PSP can improve glucose-lipid metabolism,protect cardiac function,and delay the occurrence of myocardial fibrosis in diabetic rats,and can also improve the viability of cardiomyocytes.Its mechanism of action may be related to the inhibition of cellular pyroptosis and delayed occurrence of ventricular remodeling.

The aim of this study is to explore the protective mechanism of Jiedu Tongluo Tiaogan decoction(JTTD)on islet cells of type 2 diabetic SD rats based on autophagy regulation.8-week-old SD male mice were fed with high-fat diet for 4 weeks and then induced with a single intraper-itoneal injection of streptozotocin 40 mg/kg.The Chinese medicine group was given the low(1.5 g·kg-1·d-1),medium(4.5 g·kg-1·d-1)and high(13.5 g·kg-1·d-1)doses of JTTD,and the western medicine group was given metformin hydrochloride(0.15 g·kg-1·d-1).Fasting blood glucose(FBG),glycated serum protein(GSP),triglycerides(TG),cholesterol(CHO),high-densi-ty lipoprotein cholesterol(HDL-C),low-density lipoprotein cholesterol(LDL-C),free fat(FFA)were measured.Moreover,the fasting serum insulin was determined by ELISA method and insulin resistance index(HOMA-IR)was calculated,glucose tolerance test was performed.The pancreatic tissues of SD mice were stained with HE,and the expression of Beclin-1 and LC3 were checked by immunohistochemistry,the expression of Beclin-1,LC3 and P62 was determined by Western blot,the expression of Beclin-1 and LC3RNA was determined by RT-PCR.The results showed that both the JTTD group and metformin hydrochloride group had better hypoglycemic and lipid-lowering effects(P＜0.05,P＜0.01).HE staining of pancreatic tissues showed that disorganized structure,unclear boundary,irregular arrangement,less cytoplasm,and lighter cytoplasmic staining or vacuo-lation in the islets in the JTTD group and metformin hydrochloride group were improved.In terms of autophagy regulation,immunohistochemistry,Western blot and RT-PCR,compared with the model group,the expression of Beclin-1 and LC3 mRNA was significantly increased(P＜0.01)and the level of p62 protein was decreased(P＜0.05,P＜0.01)in the JTTD group and metformin hydrochloride group.In conclusion,the JTTD can alleviate the trend of weight loss and improve ex-cessive drinking effectively in type 2 diabetic mellitus SD mice,as well as have better effects on im-proving glucose and lipid metabolism,moreover,it can alleviate the damage of pancreatic his-topathological structure and protect islet cells,which may be achieved by regulating the level of au-tophagy.

This study intends to establish a colorectal cancer(CRC)organoid model,expand and isolate CRC-reactive tumor-infiltrating lymphocytes(TILs),screen tumor-specific T cell receptors(TCRs)and perform functional verification,in order to provide a technological platform and research foundation for the clinical transformation of individualized adoptive T-cell immunotherapy for colorectal cancer.An organoid model derived from colon cancer patient tissues was constructed using in vitro 3D culture techniques,which then subjected to HE staining and immunohistochemistry for detecting morphological characteristics and representative molecular expression.Subsequently,CRC organoids were co-cultured with TILs for sorting reactive TILs using flow cytometry,and the characteristics of reactive TCR clones was analyzed through single T cell receptor gene cloning technology.Furthermore,the function of TCRs was verified through cytotoxicity experiments.Morphological analysis and representative molecules(CK20 and CDX2)expression indicated that there is high similarity between colorectal cancer organoids and patient tumors.In the in vitro expanded and cultured TILs,colorectal cancer-reactive T cells with upregulated CD137 expression and increased IFN-γ secretion were screened out successfully,among which TCR2-T cells demonstrated superior tumor reactivity and in vitro tumor killing function.In conclusion,a platform for screening and function validation of reactive TCRs based on CRC-Org has been established,providing a technological platform for the translational application of individualized T-cell therapy for colorectal cancer.

Objective:To investigate the effects of peripartum administration of low-dose corticosteroids or intravenous immunoglobulin (IVIG) on delivery outcomes in pregnant patients with primary immune thrombocytopenia (ITP).Methods:This prospective cohort study involved pregnant women (≥34 gestational weeks) who were diagnosed with ITP in Peking University People's Hospital from January 2017 to December 2021. Their platelet counts were between 20×10 9/L to 50×10 9/L without bleeding and none of them had been treated with any medications. All patients were divided into medication group (prednisone or IVIG) and platelet transfusion group based on their preference. Differences in vaginal delivery rate, postpartum hemorrhage rate and platelet transfusion volume between the two groups were compared using t-test, Wilcoxon rank sum test and Chi-square test. Binary logistic regression was used to investigate the factors influencing the rates of vaginal delivery and postpartum hemorrhage. Multiple linear regression was used to analyze the factors influencing the platelet transfusion volume. Results:A total of 96 patients with ITP were recruited with 70 in the medication group and 26 in the platelet transfusion group. The vaginal delivery rate in the medication group was higher than that in the platelet transfusion group [60.0% (42/70) vs 30.8% (8/26), χ 2=6.49, P=0.013]. After adjusted by the proportion of multiparae and the gestational age at delivery, binary logistic regression showed that the increased vaginal delivery rate in patients undergoing the peripartum treatment ( OR=4.937, 95% CI: 1.511-16.136, P=0.008). The incidence of postpartum hemorrhage in the two groups was 22.9% (16/70) and 26.9% (7/26), respectively, but no significant difference was shown ( χ 2=0.17, P=0.789). The median platelet transfusion volume was lower in the medication group than in the platelet transfusion group [1 U(0-4 U) vs 1 U(1-3 U), Z=－2.18, P=0.029]. After adjustment of related factors including the platelet count at enrollment, obstetrical complications and anemia, multiple linear regression showed that the platelet transfusion volume was also lower in the medication group (95% CI:0.053-0.911, P=0.028). Ninety-six newborns were delivered without intracranial hemorrhage. The overall incidence of neonatal thrombocytopenia was 26.0% (25/96). There was no significant difference in birth weight, and incidence of neonatal asphyxia or thrombocytopenia between the two groups. Conclusion:Peripartum therapy in ITP patients may increase vaginal delivery rate and reduce platelet transfusion volume without causing more postpartum hemorrhage.

This study was performed to explore the impact of glutamine(Gln)on the anti-tumor immune response of CD8+ T cells and its mechanism.TCGA database was used to analysis the relationship between tumor Gln metabolism and the quantity and functionality of infiltrating CD8+ T cells.CRISPR/Cas9 was employed to knock down GLS expression in mouse MC38 cells,and a mouse tumor model was established.Flow cytometry was conducted to assess tumor proliferation,apoptosis,and the quantity and functionality of tumor-infiltrating immune cells.Lymphocytes isolated from health individuals were treated with Gln-deficient media,complete media or media supplemented with GSH,RSL3 in vitro.Then the apoptosis,the expression levels of GPX4,Lipid-ROS,and effector function protein of CD8+ T cells were detected by flow cytometry.Furthermore,RNA-seq was performed to analyze the differential gene expression on the Gln-depleted CD8+ T cells.Data showed that tumor Gln metabolism was inversely associated with the quantity and functionality of tumor-infiltrating CD8+ T cells.Low expression of GLS in MC38 cells could inhibit C57BL/6 tumor growth,decrease Ki-67 expression,promote casepase-3 expression,increase the amount of tumor-infiltrating immune cells,suppress PD-1,TIM-3,and LAG-3 expression,and enhance CD137,CD107a,IFN-γ and TNF-α expression in tumor-infiltrating CD8+ T cells.RNA-seq results indicated an upregulation of ferroptosis genes TFRC,HMOX1,CYBB and SLC7A11 in CD8+ T cells following glutamine deficiency.Gln deficiency led to lower CD137,CD107a,IFN-γ,GSH,GPX4 expression,increased Lipid-ROS level,and caused cell death in CD8+ T cells.Supplementation of GSH upregulated GPX4 expression,downregulated Lipid-ROS level,and increased IFN-γ secretion in CD8+ T cells.In conclusion,Gln deficiency inhibits the effector function of CD8+ T cells by inducing ferroptosis,and promotes tumor growth.

Objective:To explore the influencing factors of the secondary sex ratio (SSR) of singleton babies born following assisted reproductive technology.Methods:A total of 7020 single babies delivered by assisted reproductive technology at the Reproductive Medicine Center of the First Affiliated Hospital of USTC (Anhui Provincial Hospital) from January 2000 to December 2018 were collected in a retrospective cohort study. The effect of influencing factors, such as maternal age, fertilization method, embryo transfer type and embryo transfer period on the sex of singleton babies were analyzed.Results:Among 7020 babies, 3730 were boys and 3290 were girls. The total SSR was 113∶100. The SSR of in vitro fertilization (IVF) was significantly higher than that of intracytoplasmic sperm injection (ICSI) fertilization (121∶100 vs. 95∶100, P<0.001), while other factors had no significant effect on the SSR of infants born. In the stratified analysis, there was no difference in SSR between IVF and ICSI in the blastocyst embryo transfer group with mother's age 35 years or older ( P<0.05), while the SSR of IVF in the other groups was higher than that of ICSI (<35 years old group: P<0.001; fresh embryo group: P=0.001; frozen-thawed embryo group: P=0.003; cleavage embryo transfer group: P<0.001). Univariate logistic regression analysis showed that the fertilization method in singleton pregnancy had a statistically significant effect on baby SSR ( OR=0.792, 95% CI=0.712-0.881, P<0.001). After removing confounding factors, logistic regression analysis showed that the fertilization method in singleton pregnancy had a statistically significant effect on neonatal SSR (a OR=0.793, 95% CI=0.713-0.883, P<0.001). Conclusion:The fertilization method of assisted reproductive technology can affect the SSR of singleton newborns. The SSR of IVF is significantly higher than that of ICSI and the difference was significant.

Objective:To explore the influencing factors of the secondary sex ratio (SSR) of singleton babies born following assisted reproductive technology.Methods:A total of 7020 single babies delivered by assisted reproductive technology at the Reproductive Medicine Center of the First Affiliated Hospital of USTC (Anhui Provincial Hospital) from January 2000 to December 2018 were collected in a retrospective cohort study. The effect of influencing factors, such as maternal age, fertilization method, embryo transfer type and embryo transfer period on the sex of singleton babies were analyzed.Results:Among 7020 babies, 3730 were boys and 3290 were girls. The total SSR was 113∶100. The SSR of in vitro fertilization (IVF) was significantly higher than that of intracytoplasmic sperm injection (ICSI) fertilization (121∶100 vs. 95∶100, P<0.001), while other factors had no significant effect on the SSR of infants born. In the stratified analysis, there was no difference in SSR between IVF and ICSI in the blastocyst embryo transfer group with mother's age 35 years or older ( P<0.05), while the SSR of IVF in the other groups was higher than that of ICSI (<35 years old group: P<0.001; fresh embryo group: P=0.001; frozen-thawed embryo group: P=0.003; cleavage embryo transfer group: P<0.001). Univariate logistic regression analysis showed that the fertilization method in singleton pregnancy had a statistically significant effect on baby SSR ( OR=0.792, 95% CI=0.712-0.881, P<0.001). After removing confounding factors, logistic regression analysis showed that the fertilization method in singleton pregnancy had a statistically significant effect on neonatal SSR (a OR=0.793, 95% CI=0.713-0.883, P<0.001). Conclusion:The fertilization method of assisted reproductive technology can affect the SSR of singleton newborns. The SSR of IVF is significantly higher than that of ICSI and the difference was significant.

Objective: To explore the effect of histone deacetylase (HDAC) inhibitor valproic acid (VPA) on the epithelial-mesenchymal transition (EMT) of colon cancer. Methods: With four colon carcinoma cell lines (DLD-1, HCT116, SW480 and HT29) as study subjects, the effect of different concentrations of VPA(0.5，5 mmol/L) on cell proliferation was detected by MTT assay. The expression level of EMT-related proteins (E-cadherin and vimentin) was detected by Western blotting; Phenotypic changes of E-cadherin and vimentin were detected by immunofluorescence staining; Cell migration and invasion ability was detected by wound healing and Transwell invasion assay, respectively. Results:After treated with different concentrations of VPAfor 48 h, low concentration of VPAmerely exerted any effect on the cell proliferation rate of four colon cancer cell lines, and thus was chosen as the experiment concentration; The results of Western blotting showed that the expression of E-cadherin was reduced (P<0.05) and vimentin was increased (P<0.05) in colon carcinoma cells by VPAtreatment (0.5 mmol/L); Immunofluorescence staining revealed membranous attenuation or nuclear translocation of E-cadherin and enhanced expression of vimentin after VPA treatment (0.5 mmol/L), and these responses occurred after 6 h and sustained until 24 h; Wound healing and Transwell invasion assay showed increased migration and invasion ability following VPA treatment (0.5 mmol/L). Conclusion: Low concentration VPA could induce the development of EMT in colon cancer cells by nuclear translocation of E-cadherin, and obviously enhance the migration and invasion ability of colon cancer cells; Thus, HDAC inhibitors, as a new type anti-cancer option, shall be carefully considered before their application in colon cancer.