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

Background::Non-alcoholic fatty liver disease (NAFLD) and inflammatory bowel disease (IBD) have shown similar worsening epidemic patterns globally and shared various overlapping pathophysiological mechanisms. However, evidence on the relationship between NAFLD and IBD risk is lacking. We aimed to investigate the associations between long-term risk of incident IBD and NAFLD in a large prospective cohort.Methods::Participants from the United Kingdom Biobank cohort (https://biobank.ndph.ox.ac.uk/) who were free of IBD and alcoholic liver disease at baseline were enrolled. Baseline non-alcoholic fatty liver degree was measured by the well-established fatty liver index (FLI). The outcomes of interest included incident IBD, ulcerative colitis (UC), and Crohn’s disease (CD). Multivariable Cox proportional hazard regression was used to calculate hazard ratios (HRs) and 95% confidence intervals (CIs).Results::Among 418,721 participants (mean FLI: 48.11 ± 30.11), 160,807 (38.40%) participants were diagnosed as NAFLD at baseline. During a median of 12.4 years’ follow-up, 2346 incident IBD cases (1545 UC, 653 CD, and 148 IBD-unclassified) were identified. Due to limited events, those IBD-unclassified were combined in UC or CD when examining the associated risk of UC or CD, separately. Compared with the lowest quartile of FLI, the highest quartile showed a separately 36.00%, 25.00%, and 58.00% higher risk of incident IBD (HR Q4 vs. Q1 =1.36, 95% CI: 1.19-1.55, Ptrend <0.001), UC (HR Q4 vs. Q1 =1.25, 95% CI: 1.07-1.46, Ptrend=0.047), and CD (HR Q4 vs. Q1 =1.58, 95% CI: 1.26-1.97, Ptrend <0.001) after multivariable adjustment. Compared with non-NAFLD, NAFLD participants had a significantly higher risk of incident IBD (HR=1.13, 95% CI: 1.04-1.24) and CD (HR =1.36, 95% CI: 1.17-1.58). Conclusions::Higher degree of non-alcoholic fatty liver is associated with increased risk of incident IBD. Interventions aimed at improving NAFLD may be a potential targeted strategy for the detection and treatment of IBD.

Hypoxia-inducible factor (HIF-1α), a core transcription factor responding to changes in cellular oxygen levels, is closely associated with a wide range of physiological and pathological conditions. However, its differential impacts on vascular cell types and molecular programs modulating human vascular homeostasis and regeneration remain largely elusive. Here, we applied CRISPR/Cas9-mediated gene editing of human embryonic stem cells and directed differentiation to generate HIF-1α-deficient human vascular cells including vascular endothelial cells, vascular smooth muscle cells, and mesenchymal stem cells (MSCs), as a platform for discovering cell type-specific hypoxia-induced response mechanisms. Through comparative molecular profiling across cell types under normoxic and hypoxic conditions, we provide insight into the indispensable role of HIF-1α in the promotion of ischemic vascular regeneration. We found human MSCs to be the vascular cell type most susceptible to HIF-1α deficiency, and that transcriptional inactivation of ANKZF1, an effector of HIF-1α, impaired pro-angiogenic processes. Altogether, our findings deepen the understanding of HIF-1α in human angiogenesis and support further explorations of novel therapeutic strategies of vascular regeneration against ischemic damage.
Humans ; Vascular Endothelial Growth Factor A/metabolism* ; Endothelial Cells/metabolism* ; Transcription Factors/metabolism* ; Gene Expression Regulation ; Hypoxia/metabolism* ; Cell Hypoxia/physiology*

AIM: To determine the patient-related risk factors for pain during phacoemulsification.METHODS: Retrospective case-control study. A total of 62 patients(62 eyes)diagnosed as cataract in the First Affiliated Hospital of Zhengzhou University from December 2023 to January 2024 were included. The numeric rating scale was used to assess the pain level within 5 min postoperatively. The highest pain value was used as the primary outcome during the procedure. Based on pain values, patients were divided into pain group(n=25)and pain-free group(n=37). Subsequently, patients in the pain group were further divided into mild(n=16), moderate(n=7), and severe groups(n=2). Spearman correlation and Logistic regression analysis were conducted to determine risk factors for pain during the phacoemulsification.RESULTS: Binary Logistic regression showed preoperative sleep durations and times of operations were important risk factors for intraoperative pain(all P<0.05). Spearman analysis showed that intraoperative pain was negatively correlated with sleep duration(rs=-0.386, P=0.002), and positively correlated with times of operations(rs=0.421, P<0.001). The results of the ordinal Logistic regression analysis showed that for every additional hour of sleep, the likelihood of experiencing one higher level of intraoperative pain decreased by 37.60%(OR=0.376, P=0.014). In contrast, the times of operations did not show a statistically significant difference(P=0.083). Receiver operating characteristic curve showed a joint prediction model of sleep duration and operative times with an area under the curve of 0.809, 84% sensitivity, and 73% specificity.CONCLUSION: The intraoperative pain during phacoemulsification is negatively correlated with sleep duration and positively correlated with times of operations.

Objective To investigate the correlation between left ventricular dysfunction and cogni-tive impairment in patients with ischemic heart disease(IHD),and explore the potential patho-genesis of cognitive impairment in IHD patients.Methods Fifty IHD patients who underwent hybrid cardiac PET/MR in our hospital from September 2020 to December 2022 were retrospec-tively enrolled in this study.According to their Montreal cognitive assessment(MoCA)scores,they were categorized into a cognitively normal group(21 cases,MoCA score ≥26)and a cogni-tively declining group(29 cases,MoCA score＜26).All patients received 13N-NH3·H2O myocar-dial perfusion imaging and 18F-FDG cardiac PET/MR to evaluate cardiac function and obtain car-diac imaging parameters.Spearman correlation analysis was applied to analyze the correlation between cardiac PET/MR parameters and MoCA scores.General linear model analysis was applied to analyze the correlation between MoCA scores and cardiac PET/MR parameters after adjusting for co variates.Results Compared with the cognitively normal group,the patients in the cognitively declining group had significantly lower stroke volume[56.95(47.51,77.64)ml vs 82.66(73.88,92.92)ml,P=0.001],stroke volume index[34.07(28.93,43.20)ml/m2 vs 44.28(38.06,49.49)ml/m2,P=0.008],cardiac output[3.92(3.18,5.34)L/min vs 5.13(4.58,5.67)L/min,P=0.007],and cardiac index[(2.42±0.68)L/(min·m2)vs(2.78±0.39)L/(min·m2),P=0.021].Spearman correlation analysis revealed that MoCA score was positively correlated with stroke volume(r=0.497,P=0.001),stroke volume index(r=0.365,P=0.009)and cardiac output(r=0.361,P=0.010).After adjusting for sex,age,education level and vascular risk factors,MoCA score re-mained positively correlated with stroke volume(r=0.497,P=0.003).Conclusion Systemic hy-poperfusion in IHD patients may be one of the mechanisms leading to cognitive impairment.

ObjectiveTo investigate the willingness of middle school students to engage in medicine and its influencing factors during COVID-19 pandemic in Lanzhou， so as to provide references for the early career planning education on middle school students. MethodsConvenient sampling method was used to choose 1 795 middle school students from 5 schools in Lanzhou from July 21 to July 31， 2022 for this study where an online questionnaire survey was distributed to collect data. A self-compiled questionnaire was used to gather the general information， assess the willingness to engage in medicine and seek the underlying reasons. The psychological resilience level of students was assessed using Resilience Scale for Adolescents. Then multivariate Logistic regression analysis was conducted to screen the factors associated with the willingness of middle school students to engage in medicine. ResultsAfter the local outbreak of COVID-19 in Lanzhou， 835 （46.52%） middle school students demonstrated their willingness to engage in medicine， and the top three reasons for their willingness were being affected by medical staff during the pandemic （75.69%）， having interests in medical sciences （67.31%）， and the potential increase of attention to national medical development after the epidemic （43.23%）. Logistical regression analysis showed that female gender （OR=1.438， P<0.01）， family members engaged in the medical and health industry （OR=1.574， P<0.01） and having a high psychological resilience score （OR=1.011， P<0.01） were related to the clear willingness to engage in medicine， while studying in senior middle school （OR=0.473， P<0.01） and excessive exposure （≥50%） to negative information during the epidemic （OR=0.797， P<0.05） were related to declining willingness to engage in medicine. ConclusionAfter the COVID-19 outbreak in Lanzhou， a large proportion of local middle school students show their willingness to engage in medicine， especially among those of female gender， having family members engaged in the medical and health industry and having high level of psychological resilience. Whereas students studying in senior middle school and with excessive exposure to negative information during the outbreak indicate low willingness to engage in medicine.

Aging poses a major risk factor for cardiovascular diseases, the leading cause of death in the aged population. However, the cell type-specific changes underlying cardiac aging are far from being clear. Here, we performed single-nucleus RNA-sequencing analysis of left ventricles from young and aged cynomolgus monkeys to define cell composition changes and transcriptomic alterations across different cell types associated with age. We found that aged cardiomyocytes underwent a dramatic loss in cell numbers and profound fluctuations in transcriptional profiles. Via transcription regulatory network analysis, we identified FOXP1, a core transcription factor in organ development, as a key downregulated factor in aged cardiomyocytes, concomitant with the dysregulation of FOXP1 target genes associated with heart function and cardiac diseases. Consistently, the deficiency of FOXP1 led to hypertrophic and senescent phenotypes in human embryonic stem cell-derived cardiomyocytes. Altogether, our findings depict the cellular and molecular landscape of ventricular aging at the single-cell resolution, and identify drivers for primate cardiac aging and potential targets for intervention against cardiac aging and associated diseases.
Aged ; Animals ; Humans ; Aging/genetics* ; Forkhead Transcription Factors/metabolism* ; Myocytes, Cardiac/metabolism* ; Primates/metabolism* ; Repressor Proteins/metabolism* ; Transcriptome ; Macaca fascicularis/metabolism*

ObjectTo observe the clinical efficacy of Huangqin Beimutang on chronic rhinosinusitis in children. MethodA randomized controlled trial （RCT） was conducted on 70 children who met the criteria for chronic rhinosinusitis， with 35 cases in the Chinese medicine group and 35 cases in the western medicine group. In the western medicine group，children received mometasone furoate nasal spray，one spray per nostril，once a day for two weeks， and also received a small dose of azithromycin suspension at 4 mg·kg^-1·d^-1，once a day，3 days a week for 2 weeks. The children in the Chinese medicine group were treated with oral Huangqin Beimutang，one dose per day for 2 weeks. Before and after treatment，the scores of primary symptoms and signs of traditional Chinese medicine （TCM），visual analogue scale （VAS） symptom scores，sinus computed tomography （CT） efficacy scores，and clinical efficacy of TCM syndromes in the two groups were evaluated，and the occurrence of adverse events was recorded. ResultThe total effective rate of clinical efficacy of TCM syndrome in the Chinese medicine group was 88.57% （31/35）， which was higher than 71.43% （25/35） in the western medicine group（χ²=8.458，P<0.05）. The VAS scores， scores of TCM primary symptoms of nasal obstruction and runny nose， and physical sign scores in both groups were lower than those after treatment （P<0.01）. The above indicators in the Chinese medicine group were superior to those in the western medicine group after treatment （P<0.05， P<0.01）. Compared with the conditions before treatment， there was no significant improvement in headache in the western medicine group， while the headache score in the Chinese medicine group decreased after treatment （P<0.01）. The CT scores of the two groups showed a downward trend， but the difference was not statistically significant. There were no adverse reactions during treatment in both groups. ConclusionHuangqin Beimutang can effectively improve the clinical symptoms of patients with chronic rhinosinusitis， and it is safe and effective.

Objective:To evaluate the value of integrated PET/MR in assessing myocardial viability in ischemic heart disease.Methods:A total of 39 patients (28 males, 11 females; age (60.1±12.0) years) diagnosed with ischemic heart disease in Xuanwu Hospital, Capital Medical University were retrospectively enrolled from September 2020 to December 2021. All patients underwent cardiac 13N-NH 3·H 2O and 18F-FDG PET/MR examinations. Late gadolinium enhancement (LGE) sequence was included in MRI scan. PET and MRI images were analyzed and myocardial viability of each myocardial segment was evaluated according to the American Heart Association (AHA) 17 segment method. The extent of left ventricular infarcted myocardium was measured based on PET and MRI images. Weighted Kappa test was used to evaluate the agreement of PET and MRI in assessing myocardial viability. The extent of infarcted myocardium measured by PET and MRI was compared by paired- t test, and Pearson correlation analysis was used to assess the correlation between them. Results:There was a moderate agreement between PET and MRI in assessing myocardial viability ( Kappa=0.532, P<0.001), with the agreement rate of 69.83%(463/663). There was no significant difference but strong correlation between the extents of infarcted myocardium measured by PET and MRI ((23.89±14.23)% vs (23.55±11.90)%; t=-0.24, P=0.809; r=0.79, P<0.001). In segments with normal perfusion and metabolism on PET, 22.52% (100/444) showed abnormal enhancement on MRI. On the other hand, 39.89% (73/183) of the segments classified as non-viable on MRI showed normal or viable on PET. Conclusion:Integrated PET/MR is able to take full advantage of the complementary nature of PET and MRI, achieving the comprehensive and accurate evaluation of myocardial viability.

Objective:To construct a mutant strain of hypervirulent Klebsiella pneumoniae NTHU-K2044 with hfq gene deletion and to analyze its biological characteristics. Methods:The hfq gene of NTUH-K2044 was knocked out by homologous recombination technology to construct △ hfq mutant strain. Its biological characteristics including growth rate, environmental stress tolerance, biofilm formation, capsular polysaccharide synthesis, resistance to neutrophil phagocytosis and lethality to Galleria mellonella larvae were analyzed by comparing with the wild-type strain using phenotypic experiments. Results:The △ hfq mutant strain of hypervirulent Klebsiella pneumoniae NTHU-K2044 was successfully constructed. Phenotypic experiments showed that the △ hfq mutant strain had significantly slower growth rate, smaller colonies and decreased hypermucoviscosity. Its growth was significantly inhibited under different environmental stress conditions such as pH9, pH5.5, 0.7 mmol/L SDS, 5% NaCl, 0.1% H 2O 2 and high temperature of 50℃. In terms of virulence and pathogenicity, the △ hfq mutant strain showed decreased ability to form biofilm and capsule, significantly down-regulated expression of magA and rmpA genes required for capsule synthesis, lower survival rate in the neutrophil bactericidal test and obviously reduced lethality to Galleria mellonella larvae. Conclusions:As a RNA chaperone, Hfq protein could participate in post-transcriptional regulation and play an important role in regulating the physiology, environmental adaptability and virulence of hypervirulent Klebsiella pneumoniae. This study provided reference for further study on hypervirulent Klebsiella pneumoniae.