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 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.

Objective:To investigate the relationship between positive asthma prediction index (API) and single nucleotide polymorphisms (SNPs) of interleukin (IL-13), IL-4, β 2 adrenergic receptor (ADRB2), and type I Fc ε receptor β (FcER1B) genes in asthmatic children.Methods:A prospective study was conducted on 102 asthmatic children under 5 years old admitted to Zhongshan Boai Hospital and Foshan First People′s Hospital (51 cases were API positive and 51 cases were API negative) from January 2020 to August 2023. Oral and buccal mucosal exfoliated cells were collected from the children, and genomic DNA was extracted using magnetic bead method. Four gene loci (IL-13 rs20541, IL-4 rs2243250, ADRB2 rs1042713, FcER1B rs569108) were genotyped using a matrix assisted laser desorption ionization time-of-flight mass spectrometer. Logistic regression analysis was used to evaluate the correlation between SNP typing at these four gene loci and API positivity in asthmatic children.Results:There was a statistically significant difference in the SNP typing and allele distribution frequency of IL-13 rs20541, IL-4 rs2243250, ADRB2 rs1042713, FcER1B rs569108 between the API positive and API negative groups of wheezing children (all P<0.05). Among API positive children, the proportion of IL-13 rs20541 site was higher in GG type, the proportion of IL-4 rs2243250 site was higher in TT type, the proportion of ADRB2 rs1042713 site was higher in AG type, and the proportion of FcER1B rs569108 site was higher in AA type; The results of logistic regression analysis showed that IL-13 rs20541 GG type, IL-4 rs2243250 TT type, FcER1B rs569108 AA type were associated with the risk of API positivity in asthmatic children (all P<0.05). Conclusions:IL-13, IL-4, and FcER1B genes are risk genes for the development of API positive wheezing in children under 5 years old. SNP typing of these genes can be used to evaluate the risk of API positivity in clinical practice.

Objective:To investigate the clinical effect of artificial intelligence(AI)technique in delineating target volume for patients with lung cancer during radiotherapy.Methods:A total of 60 patients with lung cancer who received radiotherapy in Pingxiang People's Hospital from September 2021 to March 2023 were selected,and they were divided into control group and observation group by random envelope method,with 30 cases in each group.The control group outlined target volume as conventional method.The observation group adopted deep learning technique to conduct train,and then,UNet network model was output and was used to complete automatic delineation for the target volume of radiotherapy for patients.The near-term efficacy,planning target region volume,radiation dose of target volume,volume and dose of organ at risk(OAR),survival time and incidence of adverse reactions were compared between two groups.Results:The objective relief rate(ORR)of observation group was 70.0％(21/30)after intervention,which was higher than that[46.67％(14/30)]of control group,and the difference was statistically significant(x2=5.691,P＜0.05).The radiation doses of internal target volume(ITV)and planning target volume in observation group were lower respectively than those in control group(t=4.591,4.934,P＜0.05),and the differences of them were significant,respectively.The volume percentages(V20,V5)of the exposed radiation dose that were higher than 20 Gy and 5 Gy in normal lung tissue,the exposed mean lung dose(MLD)of bilateral lungs and the exposed dose of 1cc volume(D1cc)of bilateral lungs in observation group were all lower than those in control group,the differences were statistically significant(t=5.249,4.571,6.092,5.339,P＜0.05),respectively.There was no statistical significance in the incidence of adverse reaction between two groups(P＞0.05).Conclusion:The application of AI technique in delineating target volume of radiotherapy for lung cancer can improve ORR,which is helpful to decrease the planning target volume,D95 and conformal index,and reduce the volume and dose of OAR.It does not increase the incidence of adverse reactions.