Given that ovarian stimulation is vital for assisted reproductive technology (ART) and results in elevated serum estrogen levels, exploring the impact of elevated estrogen exposure on oocytes and embryos is necessary. We investigated the effects of various ovarian stimulation treatments on oocyte and embryo morphology and gene expression using a mouse model and estrogen-treated mouse embryonic stem cells (mESCs). Female C57BL/6J mice were subjected to two types of conventional ovarian stimulation and ovarian hyperstimulation; mice treated with only normal saline served as controls. Hyperstimulation resulted in high serum estrogen levels, enlarged ovaries, an increased number of aberrant oocytes, and decreased embryo formation. The messenger RNA (mRNA)‍-sequencing of oocytes revealed the dysregulated expression of lysine-specific demethylase 6b (Kdm6b), which may be a key factor indicating hyperstimulation-induced aberrant oocytes and embryos. In vitro, Kdm6b expression was downregulated in mESCs treated with high-dose estrogen; treatment with an estrogen receptor antagonist could reverse this downregulated expression level. Furthermore, treatment with high-dose estrogen resulted in the upregulated expression of histone H3 lysine 27 trimethylation (H3K27me3) and phosphorylated H2A histone family member X (γ-H2AX). Notably, knockdown of Kdm6b and high estrogen levels hindered the formation of embryoid bodies, with a concomitant increase in the expression of H3K27me3 and γ-H2AX. Collectively, our findings revealed that hyperstimulation-induced high-dose estrogen could impair the demethylation of H3K27me3 by reducing Kdm6b expression. Accordingly, Kdm6b could be a promising marker for clinically predicting ART outcomes in patients with ovarian hyperstimulation syndrome.
Female ; Mice ; Demethylation/drug effects* ; Embryonic Stem Cells ; Estrogens/administration & dosage* ; Gene Expression/drug effects* ; Histones/metabolism* ; Jumonji Domain-Containing Histone Demethylases/metabolism* ; Mice, Inbred C57BL ; Oocytes ; Ovary/drug effects* ; Reproductive Techniques, Assisted ; Animals

BACKGROUND:Pain is the main symptom of knee osteoarthritis,which seriously affects the quality of life of elderly patients.The pain mechanism of knee osteoarthritis is complex and involves multi-tissue and multi-discipline.Visual analysis is needed to understand its cutting-edge content and research hotspots.OBJECTIVE:To analyze and summarize the cutting-edge content and research hotspot in knee osteoarthritis pain field using bibliometrics visualization software.METHODS:Literatures related to knee osteoarthritis pain from 2014 to 2024 were retrieved from Web of Science Core Collection(WoSCC).Three visualization software,Citespace,VOSviewer,and Bibliometrix R-Package were used for bibliometric and visual analysis.RESULTS AND CONCLUSION:(1)The number of articles on knee osteoarthritis pain research is increasing year by year.(2)The United States,China,and Australia were the top three countries in the number of publications in this field in 10 years.Boston University,University of Sydney,and University of Florida are the top three research institutions.OSTEOARTHRITIS AND CARTILAGE has published the most research articles in this field,and PAIN has been the most cited journal in this field.The author with the most published articles is Fillingim Roger B.The active authors in this field have formed a number of stable research teams among themselves,but there is a lack of international cooperation.(3)Daily management(motor,psychological,and intelligent management),inflammatory mechanism of pain,proteomics,combination medication,central sensitization,and pain regulation of knee osteoarthritis pain patients are the key contents of knee osteoarthritis pain research.Artificial intelligence assistance,extracorporeal shock wave therapy,radiofrequency ablation,nerve cryotherapy,intra-articular injection of blood or cell preparations may be the research frontiers in this field.

BACKGROUND:Estrogen receptor α can act as an upstream protein to regulate the expression and phosphorylation level of adenosine monophosphate-activated protein kinase(AMPK).Activation of the estrogen receptor α-AMPK signaling pathway promotes osteogenic proliferation and differentiation.OBJECTIVE:To explore the molecular mechanism of estrogen receptor α regulating AMPK and its effect on osteoblast proliferation and differentiation at osteoblast cell line and molecular biology levels.METHODS:(1)The passaged MC3T3-E1 mouse embryonic osteoblasts were divided into three groups:blank control group,mock group(transfected with pCDNA3.1 control plasmid),and estrogen receptor α group(transfected with pCDNA3.1-estrogen receptor α overexpression plasmid),and RT-qPCR and western blot methods were used to detect the hepatic kinase B1,CaMKKβ,and AMPKα1 mRNA,protein and phosphorylation levels.(2)ChIP-qPCR was used to demonstrate that estrogen receptor α interacts with the hepatic kinase B1 promoter.Dual luciferase assay was used to demonstrate that estrogen receptorα interacts with the hepatic kinase B1 promoter region to activate its transcriptional expression.(3)The cells were divided into three groups:mock+shNC group,estrogen receptor α+shNC group,and estrogen receptor α+shLKB1 group.Changes in the expression levels of hepatic kinase B1,phosphorylated hepatic kinase B1,and phosphorylated AMPKα1 proteins in the cells were detected by western blot.(4)The cells were divided into four groups:mock group,estrogen receptor α group,estrogen receptor α+5 μmol/L Compound C(AMPK inhibitor)group,and estrogen receptor α+10 μmol/L Compound C group.The expression of proteins related to the AMPK signaling pathway and related to osteogenesis and osteoinductivity were detected by western blot method.Cells were transfected for 24 hours and then subjected to osteogenic induction for 14 days.Alkaline phosphatase staining was performed and cell viability in each group was detected.Mineralized nodule formation was detected by alizarin red staining at 21 days of osteogenic induction.(5)The cells were transfected and pretreated with different concentrations of AMPK inhibitor in corresponding groups,and cell viability was detected by cell counting kit 8.RESULTS AND CONCLUSION:(1)Estrogen receptor α activates the AMPK signaling pathway in MC3T3-E1 cells.(2)Estrogen receptor α promotes liver kinase B1 transcription and mediates AMPK signaling pathway activation.(3)Estrogen receptor α promotes the proliferation and differentiation of MC3T3-E1 cells by activating the AMPK signaling pathway,and the expression of AMPKα1,p-AMPKα,osteoprotegerin,osteopontin,and Runx2 proteins was down-regulated under the intervention of AMPK inhibitor,and the viability of osteoblasts was decreased.(4)To conclude,estrogen receptor α activates the AMPK signaling pathway by acting on liver kinase B1 promoter,promotes osteoblast proliferation and osteogenic differentiation,and prevents osteoporosis.

BACKGROUND:Pain is the main symptom of knee osteoarthritis,which seriously affects the quality of life of elderly patients.The pain mechanism of knee osteoarthritis is complex and involves multi-tissue and multi-discipline.Visual analysis is needed to understand its cutting-edge content and research hotspots.OBJECTIVE:To analyze and summarize the cutting-edge content and research hotspot in knee osteoarthritis pain field using bibliometrics visualization software.METHODS:Literatures related to knee osteoarthritis pain from 2014 to 2024 were retrieved from Web of Science Core Collection(WoSCC).Three visualization software,Citespace,VOSviewer,and Bibliometrix R-Package were used for bibliometric and visual analysis.RESULTS AND CONCLUSION:(1)The number of articles on knee osteoarthritis pain research is increasing year by year.(2)The United States,China,and Australia were the top three countries in the number of publications in this field in 10 years.Boston University,University of Sydney,and University of Florida are the top three research institutions.OSTEOARTHRITIS AND CARTILAGE has published the most research articles in this field,and PAIN has been the most cited journal in this field.The author with the most published articles is Fillingim Roger B.The active authors in this field have formed a number of stable research teams among themselves,but there is a lack of international cooperation.(3)Daily management(motor,psychological,and intelligent management),inflammatory mechanism of pain,proteomics,combination medication,central sensitization,and pain regulation of knee osteoarthritis pain patients are the key contents of knee osteoarthritis pain research.Artificial intelligence assistance,extracorporeal shock wave therapy,radiofrequency ablation,nerve cryotherapy,intra-articular injection of blood or cell preparations may be the research frontiers in this field.

BACKGROUND:Estrogen receptor α can act as an upstream protein to regulate the expression and phosphorylation level of adenosine monophosphate-activated protein kinase(AMPK).Activation of the estrogen receptor α-AMPK signaling pathway promotes osteogenic proliferation and differentiation.OBJECTIVE:To explore the molecular mechanism of estrogen receptor α regulating AMPK and its effect on osteoblast proliferation and differentiation at osteoblast cell line and molecular biology levels.METHODS:(1)The passaged MC3T3-E1 mouse embryonic osteoblasts were divided into three groups:blank control group,mock group(transfected with pCDNA3.1 control plasmid),and estrogen receptor α group(transfected with pCDNA3.1-estrogen receptor α overexpression plasmid),and RT-qPCR and western blot methods were used to detect the hepatic kinase B1,CaMKKβ,and AMPKα1 mRNA,protein and phosphorylation levels.(2)ChIP-qPCR was used to demonstrate that estrogen receptor α interacts with the hepatic kinase B1 promoter.Dual luciferase assay was used to demonstrate that estrogen receptorα interacts with the hepatic kinase B1 promoter region to activate its transcriptional expression.(3)The cells were divided into three groups:mock+shNC group,estrogen receptor α+shNC group,and estrogen receptor α+shLKB1 group.Changes in the expression levels of hepatic kinase B1,phosphorylated hepatic kinase B1,and phosphorylated AMPKα1 proteins in the cells were detected by western blot.(4)The cells were divided into four groups:mock group,estrogen receptor α group,estrogen receptor α+5 μmol/L Compound C(AMPK inhibitor)group,and estrogen receptor α+10 μmol/L Compound C group.The expression of proteins related to the AMPK signaling pathway and related to osteogenesis and osteoinductivity were detected by western blot method.Cells were transfected for 24 hours and then subjected to osteogenic induction for 14 days.Alkaline phosphatase staining was performed and cell viability in each group was detected.Mineralized nodule formation was detected by alizarin red staining at 21 days of osteogenic induction.(5)The cells were transfected and pretreated with different concentrations of AMPK inhibitor in corresponding groups,and cell viability was detected by cell counting kit 8.RESULTS AND CONCLUSION:(1)Estrogen receptor α activates the AMPK signaling pathway in MC3T3-E1 cells.(2)Estrogen receptor α promotes liver kinase B1 transcription and mediates AMPK signaling pathway activation.(3)Estrogen receptor α promotes the proliferation and differentiation of MC3T3-E1 cells by activating the AMPK signaling pathway,and the expression of AMPKα1,p-AMPKα,osteoprotegerin,osteopontin,and Runx2 proteins was down-regulated under the intervention of AMPK inhibitor,and the viability of osteoblasts was decreased.(4)To conclude,estrogen receptor α activates the AMPK signaling pathway by acting on liver kinase B1 promoter,promotes osteoblast proliferation and osteogenic differentiation,and prevents osteoporosis.

Objective To analyze the correlation between high signal intensity of infrapatellar fat pad(IPFP)and symptoms and structural changes of osteoarthritis of the knee in patients with osteoarthritis of the knee based on image digitization.Methods Imaging and clinical data of patients with knee osteoarthritis(KOA)were retrospectively analyzed.The differences in knee osteoarthritis symptoms and structural changes in patients with different IPFP high signal intensities were compared,and the relationship between IPFP high signal intensity changes and knee osteoar-thritis symptoms as well as structural changes was also analyzed.Results A total of 219 patients(268 knees)were collected and completed imaging measurements and evaluations,and the median IPFP high signal intensity(27.40％)was used to compare the groups.The IPFP high signal intensity ≥ 27.40％group had a significantly higher degree of tibial cartilage damage,changes in knee angle,pain,and total scores on the osteoarthritis index(western ontario and mcMaster).universities osteoarthritis index(WOMAC)were significantly higher than those in the IPFP high signal intensity＜27.40％group,with a statistically significant difference(P＜0.05);IPFPF high signal inten-sity changes were associated with knee pain,joint line convergence angle,JLCA,and knee joint angle.Angle(JLCA),knee kellgren-Lawrence(KL)classification,and the degree of femoral and tibial cartilage damage were significantly positively correlated(P＜0.05).Conclusion In patients with KOA,there is a significant correlation between IPFP high signal intensity changes and knee pain symptoms as well as structural changes,which may be an important factor influencing the progression of KOA.

Objective To analyze the correlation between high signal intensity of infrapatellar fat pad(IPFP)and symptoms and structural changes of osteoarthritis of the knee in patients with osteoarthritis of the knee based on image digitization.Methods Imaging and clinical data of patients with knee osteoarthritis(KOA)were retrospectively analyzed.The differences in knee osteoarthritis symptoms and structural changes in patients with different IPFP high signal intensities were compared,and the relationship between IPFP high signal intensity changes and knee osteoar-thritis symptoms as well as structural changes was also analyzed.Results A total of 219 patients(268 knees)were collected and completed imaging measurements and evaluations,and the median IPFP high signal intensity(27.40％)was used to compare the groups.The IPFP high signal intensity ≥ 27.40％group had a significantly higher degree of tibial cartilage damage,changes in knee angle,pain,and total scores on the osteoarthritis index(western ontario and mcMaster).universities osteoarthritis index(WOMAC)were significantly higher than those in the IPFP high signal intensity＜27.40％group,with a statistically significant difference(P＜0.05);IPFPF high signal inten-sity changes were associated with knee pain,joint line convergence angle,JLCA,and knee joint angle.Angle(JLCA),knee kellgren-Lawrence(KL)classification,and the degree of femoral and tibial cartilage damage were significantly positively correlated(P＜0.05).Conclusion In patients with KOA,there is a significant correlation between IPFP high signal intensity changes and knee pain symptoms as well as structural changes,which may be an important factor influencing the progression of KOA.