BACKGROUND: Knee osteoarthritis (OA) is still challenging to prevent or treat. Enhanced endoplasmic reticulum (ER) stress and increased pyroptosis in chondrocytes may be responsible for cartilage degeneration. This study aims to investigate the effect of ER stress on chondrocyte pyroptosis and the upstream regulatory mechanisms, which have rarely been reported. METHODS: The expression of the histone methyltransferase enhancer of zeste homolog 2 (EZH2), microRNA-142-3p (miR-142-3p), and high mobility group box 1 (HMGB1) and the levels of ER stress, pyroptosis, and metabolic markers in normal and OA chondrocytes were investigated by western blotting, quantitative polymerase chain reaction, immunohistochemistry, fluorescence in situ hybridization, fluorescein amidite-tyrosine-valine-alanine-aspartic acid-fluoromethyl ketone (FAM-YVAD-FMK)/Hoechst 33342/propidium iodide (PI) staining, lactate dehydrogenase (LDH) release assays, and cell viability assessments. The effects of EZH2, miR-142-3p, and HMGB1 on ER stress and pyroptosis and the hierarchical regulatory relationship between them were analyzed by chromatin immunoprecipitation, luciferase reporters, gain/loss-of-function assays, and rescue assays in interleukin (IL)-1β-induced OA chondrocytes. The mechanistic contribution of EZH2, miR-142-3p, and HMGB1 to chondrocyte ER stress and pyroptosis and therapeutic prospects were validated radiologically, histologically, and immunohistochemically in surgically induced OA rats. RESULTS: Increased EZH2 and HMGB1, decreased miR-142-3p, enhanced ER stress, and activated pyroptosis in chondrocytes were associated with OA occurrence and progression. EZH2 and HMGB1 exacerbated and miR-142-3p alleviated ER stress and pyroptosis in OA chondrocytes. EZH2 transcriptionally silenced miR-142-3p via H3K27 trimethylation, and miR-142-3p posttranscriptionally silenced HMGB1 by targeting the 3'-UTR of the HMGB1 gene. Moreover, ER stress mediated the effects of EZH2, miR-142-3p, and HMGB1 on chondrocyte pyroptosis. In vivo experiments mechanistically validated the hierarchical regulatory relationship between EZH2, miR-142-3p, and HMGB1 and their effects on chondrocyte ER stress and pyroptosis. CONCLUSIONS A novel EZH2/miR-142-3p/HMGB1 axis mediates chondrocyte pyroptosis and cartilage degeneration by regulating ER stress in OA, contributing novel mechanistic insights into OA pathogenesis and providing potential targets for future therapeutic research.
Enhancer of Zeste Homolog 2 Protein/genetics* ; Osteoarthritis, Knee/pathology* ; Chondrocytes/metabolism* ; Pyroptosis/physiology* ; HMGB1 Protein/genetics* ; MicroRNAs/metabolism* ; Endoplasmic Reticulum Stress/genetics* ; Humans ; Animals ; Rats ; Male ; Rats, Sprague-Dawley ; Middle Aged

Objective To find the molecular mechanism underlying the effect of congenital cataract related 129th single-site mutation G129C on the thermal stability of human γC crystallin(HγC)protein structure.Methods HγC-WT and HγC-G129C were expressed and purified in vitro.The changes of intrinsic fluorescence intensity and static light scattering intensity of proteins with temperature were measured,and the temperature dependence of the folding and aggregation structures of HγC-WT and HγC-G129C was compared.Results When temperature was be-low 65 ℃,the barycentric mean of the intrinsic fluorescence of HγC-WT and HγC-G129C shifted towards a longer wavelength and the fluorescence intensity decreased with the increasing temperature,which was believed to be the evidence of unfolded protein conformation.When the temperature was higher than 65 ℃,the static light scattering intensity increased significantly with the temperature,indicating the protein aggregation upon heating.The wild-type HγC-G129C showed a stronger aggregation potency.During the thermal de-naturation process of HγC-WT and HγC-G 129C,the crossing-point temperatures were 74.5 ℃ and 55.5 ℃,respectively.HγC-WT showed higher thermal stability.Conclusions The congenital cataract-associated G129C mutation significantly weakens conforma-tional stability of γC-crystallin.

In recent years， the incidence of pulmonary nodules has kept rising. To give full play to the advantages of traditional Chinese medicine （TCM） in the treatment of pulmonary nodules and identify the breakthrough points of integrating TCM with Western medicine， the China Association of Chinese Medicine organized medical experts in TCM and western medicine to carry out in-depth discussion regarding this disease. The discussion encompassed the modern medical advances， TCM theories of etiology and pathogenesis， the role and advantages of TCM in the whole course management of pulmonary nodules， contents and methods of research on pulmonary nodules， and science popularization work， aiming to provide a reference for clinical practice and scientific research. After discussion， the experts concluded that the occurrence of pulmonary nodules was rooted in the deficiency of the lung and spleen and triggered by phlegm dampness， blood stasis， and Qi stagnation. TCM can treat pulmonary nodules by controlling and reducing nodules， improving physical constitution， ameliorating multi-system nodular diseases， reducing anxiety and avoiding excessive diagnosis and treatment， and serving as an alternative for patients who are unwilling or unfit for surgical treatment. At present， the optimal diagnosis and treatment strategy for pulmonary nodules has not been formed， which needs to be further studied from multiple perspectives such as clinical epidemiology， biology， and evidence-based medicine. The primary task of current research is to find out the advantages， effective prescriptions， and target populations and determine the effective outcomes of TCM in the treatment of pulmonary nodules. At the same time， basic research should be carried out to explore the etiology and biological behaviors of pulmonary nodules. The expert consensus on the diagnosis and treatment of pulmonary nodules with integrated TCM and Western medicine needs to be continuously revised to guide clinicians to conduct standardized， scientific， and accurate effective diagnosis and treatment.

Purpose To explore the expression,mechanism and clinical significance of MCM5 in ovarian cancer.Methods The expression of MCM5 mRNA in ovarian cancer and its correlation with patients'survival were analyzed using GEO and TCGA databases.The expression of MCM5 protein in ovarian cancer was detected by immunohistochemistry of SP two-step method,and its relationship with clinicopathological characteris-tics was analyzed.With inhibition of MCM5 by siRNA in ovarian cancer cells.The effects of MCM5 on cell proliferation,migra-tion,invasion,and apoptosis were detected by CCK-8 assay,EDU,plate cloning,Transwell chamber and flow cytometry.Re-sults Immunophenotype:MCM5 does not stain in the fallopian tube epithelium(0/6),with a positivity rate of 48.3％(57/118)in ovarian cancer.The expression of MCM5 in ovarian cancer is significantly higher than in fallopian tube epithelium,showing diffuse strong expression in high-grade serous carcino-ma.MCM5 expression is strongly correlated with ER-negative status and high Ki67 proliferation index.Knocking down MCM5 expression inhibits proliferation(P＜0.05),clonogenicity(P＜0.05),invasion and migration(P＜0.05)of ovarian cancer cells,and promotes apoptosis.Conclusion MCM5 is highly expressed in human ovarian cancer cells and tissues and is asso-ciated with poor prognosis.It is expected to become a new target for the treatment of ovarian cancer.

Objective:To evaluate the effects of different imaging frequencies and matching strategies of cone-beam computed tomography (CBCT) on dose-volume parameters in target and organs at risk (OAR) during image-guided radiotherapy for prostate cancer.Methods:A total of 561 sets of CBCT images from 21 patients treated with radical prostate radiotherapy who were admitted to Peking University First Hospital from June 2022 to May 2023 were retrospectively analyzed. All patients received volumetric intensity modulated arc therapy (VMAT) at a prescribed dose of 70 Gy divided into 25 times, 2.8 Gy per time. Clinical target volume (CTV) and OAR were delineated by the same oncologist on each CBCT image. The planned CT (pCT) was rigorously registered to CBCT after calibration of positioning errors according to different image guidance modes and frequencies, and CT values and structures were propagated to CBCT through deformable image registration (DIR). The daily dose was mapped to pCT according to the deformation vector field (DVF) for dose accumulation. The actual cumulative dose of daily online CBCT validation was compared with the weekly CBCT validation regimen (days 1, 2, 3, 6, 11, 16 and 21 online imaging). The dosimetric comparison was also made between bone-based matching and soft tissue-based matching (after automatic bone-based matching, manual prostate-based matching was performed and fine-tuning was made regarding the anterior wall of rectum). Wilcoxon signed rank-sum test was utilized to analyze dose-volume parameters between planned and cumulative doses that exhibited non-normal distribution, while paired t-test was employed for assessing shift values and average dose parameters that demonstrated normal distribution. Results:Compared with daily CBCT image guidance, the CTV_D 98% in weekly CBCT was significantly reduced [(69.08±1.58) vs. (65.24±3.64) Gy, P<0.001]. The CTV_D 98% of bone-based matching was (69.27±2.14) Gy, but the high-dose volume of the rectum were significantly increased: V 60 Gy was 3.18%±3.10%, V 65 Gy was 0.77%±1.23%. The target area coverage using soft tissue-based matching is sufficient, with a CTV_D 98% of (69.08±1.58) Gy. And the percentage volume of high-dose volume of the rectum was significantly reduced, with V 60 Gy being 2.02%±2.42% and V 65 Gy being 0.34%±0.68%. Conclusions:In prostate cancer patients undergoing moderately-fractionated radiotherapy, daily CBCT image guidance demonstrates superior target coverage compared to a weekly scheme. Soft tissue-based matching, which is automatic bone-based matching followed by manual soft tissue-based matching and fine-tuning according to the anterior rectal wall, offers better rectal protection while maintaining target coverage.

Objective To explore the effect of miRNA-222-3p in endothelial progenitor cell-derived exosomes(EPCs-Exo)on skin wound healing in diabetic mice.Methods Endothelial progenitor cell(EPCs)derived from C57BL/6 mouse bone marrow were identified by fluorescence staining.Subsequently,EPCs-Exo isolated from the media of EPCs were identified,and high-throughput sequencing of EPCs-Exo miRNA was completed.The skin injury model of diabetic mice was established.According to different groups,the wounds were treated with externally ap-plied phosphate buffer solution(PBS)and EPCs-Exo;Subcutaneous injection of PBS,agomiR-222-3p,and an-tagomiR-222-3p at the edge of the wound was performed continuously for 14 days,and the wound healing rate was observed.Meanwhile,immunofluorescence methods were used to investigate the changes in the expression levels of ROS,CD31,and VEGF in the wound margin tissue before and after treatment.Results EPCs-Exo significantly ac-celerated skin wound healing in diabetic mice,reduced the level of ROS,and increased the expression level of VEGF and CD31 in the wound margin tissue(P＜0.05).High-throughput sequencing showed that miRNA-222-3p was highly expressed in EPCs-Exo.Local subcutaneous injection of miRNA-222-3p into wound margin tissue signifi-cantly promoted the skin wound healing of diabetic mice,reduced the level of ROS,and increased the expression level of VEGF and CD31 in the wound margin tissue(P＜0.05).Conclusion MiRNA-222-3p promotes wound healing in diabetic mice and plays an important role in EPCs-Exo promoting wound healing.

Objective: To explore the correlation between Diabetes Complication Severity Index(DCSI), C-reactive protein/albumin ratio(CAR) and death in patients with diabetic foot ulcer(DFU) and to clarify their predictive value for all-cause death in DFU patients. Methods: Retrospectively analyzed the clinical data of 354 DFU patients who were treated in the Endocrinology Department of the First Affiliated Hospital of Anhui Medical University from July 2019 to December 2022. Based on survival status during follow-up, patients were divided into a survival group(n=268) and a death group(n=86). Univariate and multivariate Cox regression analyses were used to identify risk factors for all-cause death in DFU patients. Receiver operating characteristic(ROC) curves were plotted to evaluate the predictive value of DCSI, CAR, and their combination for all-cause death in DFU patients. Kaplan-Meier curves were used to explore the impact of different DCSI and CAR levels on survival in DFU patients. Results: Univariate Cox regression analysis showed that older age, history of hypertension, higher Wagner classification levels, and elevated levels of CRP, Scr, FDP, DCSI score, and CAR were associated with a higher risk of death in DFU patients(P<0.05). Higher levels of HGB, HCT, ALB, or eGFR were associated with a lower risk of death. Patients receiving combined insulin and oral hypoglycemic medication had a lower risk of death compared to those receiving only insulin therapy(P<0.05). Multivariate Cox regression analysis indicated that older age, higher levels of Scr, DCSI, and CAR were independent risk factors for all-cause death in DFU patients, while higher levels of ALB and combined insulin and oral hypoglycemic therapy were protective factors. ROC curve analysis showed that the AUC values for DCSI, CAR, and their combination were 0.652, 0.633, and 0.686, respectively. Kaplan-Meier curve analysis revealed that patients with high DCSI scores(≥4.5) had a lower survival rate compared to those with lower DCSI scores(<4.5). Similarly, patients with high CAR levels(≥0.124) had a lower survival rate compared to those with lower CAR levels(<0.124). Conclusion High levels of DCSI and CAR are independent risk factors for all-cause death in DFU patients. DCSI, CAR, and their combination have predictive value for all-cause mortality in DFU patients.

Objective To investigate the correlation between thyroid function,neutrophil/lymphocyte ratio(NLR),platelet/lymphocyte ratio(PLR)and diabetic ketoacidosis(DKA).Methods A total of 272 diabetic patients admitted to the Department of Endocrinology,Anhui No.2 Provincial People's Hospital were enrolled in this study from January 2021 to March 2024 and divided into DKA group(n=74),diabetic ketosis group(DK group,n=100)and simple DM group(n=98)according to their disease conditions.Serum free triiodothyronine(FT3),free tetraiodothyronine(FT4),thyroid stimulating hormone(TSH),FPG,HbA1c,anion gap(AG),white blood cells(WBC),neutrophils(N),platelets(PLT)and lymphocytes(L)were detected in each group,and NLR and PLR were calculated.The influence factors of DKA were analyzed by logistic regression,and the diagnostic value of FT3,NLR and PLR in DKA was evaluated by receiver operating characteristic(ROC)curve.Results Hospitalization days,FPG,HbA1c,incidence of LT3S,AG,WBC,N,PLT,NLR,PLR were higher in DKA group than in DM and DK groups(P<0.05),Age,FT3,FT4,TSH,L were lower in DKA group than in DM and DK groups(P<0.05).Logistic regression analysis showed that FT3,AG,NLR and PLR were the influencing factors for DKA.ROC curve showed that the area under the curve of FT3,NLR,PLR and combined detection to predict DKA were 0.961,0.938,0.810 and 0.980,the sensitivity was 87.8%,90.5%,75.7%and 98.6%,the specificity was 95.5%,92.9%,76.3%,92.4%,and the combined detection was superior to the single detection.Conclusions FT3,NLR,PLR have a certain diagnostic value for DKA,and the combined detection of the three has a high diagnostic value,which can provide a reference for clinical diagnosis of DKA.

Objective To establish a risk prediction model for type 2 diabetes mellitus(T2DM)related overactive bladder(OAB),and to verify the diagnostic efficacy and clinical application value of the model.Methods The clinical data of 298 patients with diabetes who underwent urodynamic examination in The First Affiliated Hospital of Wannan Medical College from January 2020 to October 2023 were analyzed retrospectively.According to the results of urodynamics,all the patients were divided into T2DM group(T2DM,n=218)and OAB group(OAB,n=80).The risk factors of OAB were analyzed by logistic regression,and the prediction model was established according to the risk factors.Receiver operating characteristic(ROC)curve,calibration diagram and decision curve analysis(DCA)were used to analyze and evaluate the diagnostic efficiency of the model.Results Logistic regression analysis results showed that age,DM duration,BMI,HbA1c,DPN and OAB score(OABSS),were independent predictors for OAB.The nomogram model built based on the above risk factors had good predictive ability,and its area under ROC curve was 0.937.The correction curve showed that the predicted probability of OAB in patients with type 2 diabetes mellitus was basically parallel to the actual probability of urodynamic detection.The results of DCA show that the model has high clinical value.Conclusions The nomogram model built by age,DM duration,OABSS,BMI,HbA1c,DPN has high predictive efficacy for OAB in patients with T2DM.

Objectives:To investigate the effect of body mass index(BMI)on perioperative and long-term prognosis of patients with severe aortic stenosis(AS)after transcatheter aortic valve replacement(TAVR). Methods:This retrospective study imcluded 180 patients with severe AS who received TAVR in Fujian Provincial Hospital from January 2019 to January 2022.According to the BMI,patients were divided into four groups:low weight group(BMI<18.5 kg/m2,n=23),normal weight group(18.5 kg/m2≤BMI<24.0 kg/m2,n=65),overweight group(24.0 kg/m2≤BMI<28.0 kg/m2,n=57),obesity group(BMI≥28.0 kg/m2,n=35).The general clinical characteristics,imaging parameters,perioperative indexes,all-cause death and the incidence of other adverse cardiac events during(18.0±6.8)months follow-up were compared among different groups.Risk factors for the perioperative complications and long-term outcomes of TAVR were evaluated. Results:The prevalence of hypertension and diabetes,left ventricular end-diastolic diameter,ventricular septal thickness and left ventricular posterior wall thickness were significantly higher in the obese group than in normal weight group(all P<0.05).The level of prealbumin in low weight group was lower than in normal weight group(P<0.05).The total perioperative complications in low weight group were higher than in normal weight group(60.9%vs.12.3%,P=0.042).During(18.0±6.8)months follow-up,the incidence of all-cause death in the low weight group was significantly higher than that in normal weight group,overweight group and obese group(17.4%vs.4.6%vs.3.5%vs.5.7%,P=0.003).Kaplan-Meier survival analysis evidenced higher mortality rate in low weight group at 18 months after TAVR(log-rank P<0.01).Multivariate Cox regression analysis showed that the risk of long-term adverse cardiovascular events was significantly higher in low weight group than in normal weight group(HR=7.633,95%CI:1.012-57.564,P=0.049). Conclusions:Low weight patients with severe AS have a higher incidence of perioperative complications and a poor long-term prognosis.Such patients should appropriately strengthen their nutritional intake and adjust their body weight to normal levels before performing TAVR.