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

To develop a method for determining the antibody-dependent cell-mediated phagocytosis (ADCP) activity of human epidermal growth factor receptor 2 (HER2)-targeted antibody drug conjugate (ADC) based on the reporter gene assay, we established an ADCP activity assay with Jurkat/NFAT/FcγRIIa cells as the effector cells and BT474 as the target cells. Then, the target cell density, the ratio of effector to target cells, the target cell adhesion time, the incubation time for drug administration, and the induction time after adding effector cells were optimized by the method of design of experiment (DOE). The method showed a significant dose-response relationship, which was complied with the four-parameter equation: y=(A-D)/[1+(x/C)^B]+D. The durability ranges of the target cell density, the ratio of effector to target cells, the target cell adhesion time, the incubation time for drug administration, and the induction time after adding effector cells were (2.5-4.0)×10⁵ cells/mL, 3-5, 1.0-2.0 h, 0 h, and 5.0-6.0 h, respectively. The results of the methodological validation showed that the linear equation was y=1.106 8x-0.011 6, r=0.969 2. The established method showed the relative accuracy ranging from -6.59% to 2.98% and the geometric coefficient of variation less than 11% in the intermediate precision test. Furthermore, the method was target-specific. The method was then applied to the determination of ADCP activity of HER2-targeted ADC, demonstrating the result of (103.5±5.7)%. We developed a reporter gene assay for determining the ADCP activity of HER2-targeted ADC and the assay demonstrated high accuracy and good reproducibility, which proposes a highly efficient and approache for evaluating ADCP effect of this HER2-targeted ADC, and also provides a referable technique for characterizing the Fc effector functions of ADCs with diverse targets.
Humans ; Receptor, ErbB-2/immunology* ; Phagocytosis/drug effects* ; Immunoconjugates/immunology* ; Genes, Reporter ; Antibody-Dependent Cell Cytotoxicity ; Jurkat Cells

Objective To explore the predictive value of heparin-binding protein(HBP)combined with soluble growth-stimulated gene 2 protein(sST2)for the prognosis of elderly patients with acute heart failure(AHF).Methods A total of 338 elderly AHF patients who were treated in the Affiliated Hospital of Xuzhou Medical University and Kuitun Hospital of Ili Kazakh Autonomous Prefecture from April 2021 to March 2022 were selected as the research subjects.The patients were followed up for 1 year.According to whether the ma-jor adverse cardiovascular and cerebrovascular events(MACE)occurred during the follow-up period,they were divided into the MACE group and the non-MACE group,respectively.The baseline characteristics of the patients were recorded,laboratory indicators were detected,transthoracic color Doppler ultrasound examina-tion was completed,the risk factors affecting the prognosis of elderly patients with AHF,as well as the predic-tive value of HBP and sST2 for the prognosis of elderly patients with AHF were analyzed.Results Among the 338 elderly AHF patients,101 patients experienced MACE,and 237 patients did not.The levels of serum creatinine,NT-proBNP,troponin T,white blood cell count(WBC),left ventricular end-diastolic diameter(LVEDD),HBP,and sST2 in the MACE group were higher than those in the non-MACE group,while the left ventricular ejection fraction(LVEF)was lower than that in the non-MACE group,and the differences were statistically significant(P＜0.05).LVEF was a protective factor for the occurrence of MACE(HR＜1,P＜0.05)and age,diabetes mellitus,creatinine,NT-proBNP,troponin T,LVEF,WBC,HBP,and sST2(HR≥1,P＜0.05)were the influencing factors for the occurrence of MACE.The optimized model(Model 5)has the highest predictive efficiency for the prognosis of elderly patients with AHF,with an area under the curve(AUC)of 0.976(95％CI:0.962-0.990).Conclusion Both HBP and sST2 are independent risk factors for the occurrence of MACE in elderly AHF patients.The predictive model established by combining HBP and sST2 has a high predictive value.

Objective:To investigate the current application of blood purification in the treatment of acute poisoning within Jiangsu Province and to evaluate the impact of extracorporeal blood purification on the clinical outcomes of critically poisoned patients.Methods:This multicenter, cross-sectional real-world observational study followed patients presenting with poisoning to the emergency departments of nine hospitals in Jiangsu Province between June 2015 and May 2019. Data were collected on demographic characteristics, vital signs within the first hour of emergency presentation, treatment modalities, length of hospital stay, and survival outcomes. Clinical data from patients who underwent extracorporeal blood purification were compared with those who did not, using the Wilcoxon rank-sum test and Chi-square test.Results:A total of 4 178 poisoning cases were included between June 2015 and May 2019. Among them, 21.7% (908/4 178) received blood purification therapy, while 78.3% (3 270/4 178) did not. Hemoperfusion (90.4%) was the most frequently employed method, followed by continuous renal replacement therapy (CRRT) (4.4%). In combined blood purification modalities, 4.8% underwent hemoperfusion combined with CRRT, 0.1% received hemoperfusion with plasma exchange, and another 0.1% underwent hemoperfusion combined with both CRRT and plasma exchange. Among patients who underwent blood purification, pesticide poisoning was the most prevalent (76.3%), with the most common toxic agents being paraquat (23.7%), dichlorvos (8.7%), methamidophos (5.2%), omethoate (4.0%), and glyphosate (3.7%). Compared to the non-blood purification group, patients in the blood purification group were more likely to present within the first hour with a low Glasgow Coma Scale (GCS) score (3-8) (22.6% vs. 9.7%, P <0.05), low mean arterial pressure (8.0% vs. 3.2%, P <0.05), longer hospital stays [5(3,9) days vs. 2(1,4) days, P <0.05] and a higher in-hospital mortality rate (21.1% vs. 5.3%, P <0.05). Follow-up via telephone 28 days after discharge revealed a survival rate of 78.9%, with a mortality rate of 21.1% in the blood purification group. Conclusions:Hemoperfusion is the most commonly utilized blood purification technique for treating poisoning in Jiangsu Province, with pesticides being the primary toxic agents treated. Although the mortality rate is higher in the blood purification group, the intervention may still contribute to improved patient outcomes.

ObjectiveTo identify the active components in Maimendong decoction (MMDD) against pulmonary fibrosis (PF) and validate their molecular effects in vitro, while focusing on the role of methylophiopogonanone B in regulating fibrosis.MethodsData on MMDD components and targets were gathered from databases including BATMAN-TCM and PubMed, whereas the PF gene data were sourced from GeneCards, OMIM, and TTD. Shared targets were determined using the STRING database, and molecular docking was used to analyze the essential molecules associated with fibrosis. To simulate PF conditions, human embryonic lung fibroblasts (HPF) and A549 cells were exposed to transforming growth factor-β1 (TGF-β1). Various assays were used to determine the effects of MMDD and methylophiopogonanone B on signaling pathways, apoptosis, and epithelial–mesenchymal transition.ResultsWe identified 11 active components from MMDD extracts that targeted 511 shared proteins associated with PF, revealing 10 key targets in network analysis. Gene ontology analysis indicated that processes and pathways such as apoptosis regulation and PI3K/Akt signaling were involved. In vitro experiments revealed that MMDD downregulated the expression of α-smooth muscle actin (α-SMA), collagen type I (COL-I), and collagen type III and regulated Bcl-2/Bax signaling pathways to promote apoptosis. The flow cytometry apoptosis assay revealed that MMDD promoted the TGF-β1-induced apoptosis of myofibroblasts. The primary active ingredient in MMDD, methylophiopogonanone B, reduced α-SMA, COL-I, and PI3K/Akt/mTOR-related protein levels in TGF-β1-treated HPF cells, decreased Bcl-2 and cleaved caspase 3, and increased Bax. Moreover, methylophiopogonanone B increased E-cadherin levels and reduced α-SMA, fibronectin, N-cadherin, vimentin, and snail in TGF-β1-treated A549 cells.ConclusionMethylophiopogonanone B demonstrated the potential to treat PF by inducing myofibroblast apoptosis and inhibiting EMT. However, despite encouraging initial results, further clinical research is warranted to verify the safety and efficacy of methylophiopogonanone B in the management of PF

Objective:To investigate the current situation and related factors of children's growth and development in drinking-water-borne endemic fluorosis areas after water improvement, and provide a basis for improving the control strategies.Methods:The stratified random sampling method was used to select children aged 7 to 13 in villages with different years of water improvement in Xi'an from November 2019 to June 2020. The height and weight of children were measured, and body mass index (BMI) was calculated. The height development level of children was graded according to the "Standard for Height Level Classification among Children and Adolescents Aged 7 - 18 Years" (WS/T 612-2018), and the physical development (BMI) level of children was determined according to the "Comprehensive Evaluation of Development Level for Children and Adolescents" (GB/T 31178-2014). At the same time, according to the standard of "Diagnosis of Dental Fluorosis" (WS/T 208-2011), the diagnosis of dental fluorosis in children was performed. The serum fluoride level was determined by ion selective electrode method, and the levels of other chemical elements (calcium, iron, magnesium, copper, zinc, iodine, selenium, lead, arsenic, cadmium, chromium, mercury, nickel) in blood were determined by inductively coupled plasma mass spectrometry.Results:A total of 469 children were included in the investigation. After water improvement, 67.38% (316/469) of the children in the disease area were at the moderate level of height development, 7.89% (37/469) were at the medium to upper and upper levels, and 24.73% (116/469) were at the medium to lower and lower levels. The BMI of the children in the emaciation group was 8.53% (40/469), while it was 10.87% (51/469) in the overweight group and 8.74% (41/469) in the obesity group. There was statistically significant difference in the distribution of height development level of children in areas with different water improvement years ( P = 0.005), but no statistically significant difference in BMI distribution ( P = 0.154). There was no significant difference in height development level and BMI distribution of children with or without dental fluorosis ( P > 0.05). There were significant differences in serum levels of iron and zinc among children with different height development levels ( P < 0.05). There were significant differences in serum levels of magnesium, copper, iodine and chromium among children with different BMI ( P < 0.05). Conclusion:The growth and development of children in drinking-water-borne endemic fluorosis area after water improvement are not correlated with the prevalence of dental fluorosis, but may be related to the levels of chemical elements such as iron, magnesium, copper, and iodine in the body.

Objective:To investigate the indoor radon concentrations and to analyze their seasonal variations in urban and rural residential dwellings in Ningxia province.Methods:From March 2022 to March 2023, based on the administrative region division and population distribution in Ningxia, a total of 143 typical residential dwellings, including 82 urban houses and 61 rural houses, were selected to measure indoor radon concentration by CR-39 solid nuclear track detectors for 1 year in Ningxia, with detectors changed every 3 months.Results:The annual average indoor radon concentration in Ningxia was 88 Bq/m 3, range 39-226 Bq/m 3. The annual average indoor radon concentration was below 100 Bq/m 3 for 69.9% of the measured dwellings, and below 300 Bq/m 3 for all of surveyed dwellings. Indoor radon concentrations in rural areas were much higher than those in urban areas ( Z=5.85, P<0.05). Indoor radon concentration varied significantly with the seasons, higher in autumn and winter, but lower in spring and summer, in total ( χ2=63.97, P<0.05), urban ( χ2=24.74, P<0.05), and rural ( χ2=43.15, P<0.05). Conclusion:The annual average indoor radon concentrations in all the measured dwellings are below the reference level of 300 Bq/m 3 recommended by Indoor Air Quality Standard (GB/T 18883-2022) in China.

Objective:To investigate the safety and efficacy of ultrasound-guided percutaneous radiofrequency ablation in the treatment of plasma cell mastitis.Methods:A retrospective analysis was performed for the clinical data of 10 patients with plasma cell mastitis treated with ultrasound-guided percutaneous radiofrequency ablation in the First Affiliated Hospital of Zhengzhou University from January 2021 to December 2022. The postoperative complications, symptoms and signs of the patients, ultrasound imaging manifestations, treatment efficacy at 3 months after surgery, the reduction rate of ablation lesions at 1 month, 3 months, 6 months, and 12 months after surgery, the recurrence status within 1 year after surgery, and the satisfaction with the treatment effect were observed.Results:The reduction rates of ablation lesions at 1 month, 3 months, 6 months, and 12 months were 32.31%-51.00%[(42.78±4.48)%], 70.66%-86.68%[(70.45±12.43)%], 72.31%-100%[(86.91±7.45)%], 89.13%-100%[(96.07±7.45)%], respecctively. The symptoms and signs of 10 patients improved significantly 3 months after surgery, and there was no recurrence of the disease during the follow-up period, and the treatment effect was significant and satisfactory.Conclusions:Ultrasound-guided percutaneous radiofrequency ablation for the treatment of plasma cell mastitis is a minimally invasive, effective and safe treatment.