Objective To explore the correlations of the severity degrees of coronary stenosis with cardiac function and exercise tolerance. Methods A total of 112 patients who underwent coronary angiography in Cardiology Department of Zhongshan Hospital, Fudan University between October 2024 and January 2025 were enrolled. According to the imaging results, the Gensini score was calculated, and the patients were divided into three groups based on the scores: mild group (＜20 points, n＝42), moderate group (20-40 points, n＝43), and severe group (＞40 points, n＝27). The left ventricular ejection fraction (LVEF), N-terminal pro-brain natriuretic peptide (NT-proBNP) level and its abnormal elevation ratio, 6-minute walk distance (6MWD), and grip strength were compared among the groups. The correlations between Gensini score and various indicators were analyzed using multivariate linear regression. And the multivariate binary logistic regression analysis was used to analyze the related factors of severe coronary stenosis. Results The 6MWD and LVEF values in the severe group were lower than those in the mild and moderate groups (P＜0.01), while the NT-proBNP level and its abnormal elevation ratio in the severe group were higher than those in the mild group (P＜0.05). The Gensini score was negatively correlated with 6MWD (β＝−0.965, P＜0.01), and positively correlated with NT-proBNP level and its abnormal elevation ratio (β＝0.015, P＜0.001; β＝0.003, P＝0.037). 6MWD and diabetes were independent related factors of severe coronary stenosis (OR＝0.956, P＜0.001; OR＝5.701, P＝0.038). Conclusions The cardiac function and exercise tolerance in patients with severe coronary stenosis decrease, 6MWD is helpful of recognizing severe coronary stenosis population.

Objective: To investigate the effect of the senescence state of parental human bone marrow mesenchymal stem cells (BMSCs) on the osteoinductive properties of their derived apoptotic vesicles (apoVs), and to provide an experimental basis and a quality control reference for the treatment of alveolar bone defects and osteoporosis based on apoVs. Methods: A replicative senescence model of human aging BMSCs (A-BMSCs) was established via serial passaging, with young BMSCs (Y-BMSCs) serving as controls. Following the induction of apoptosis with staurosporine, apoVs were isolated from Y-BMSCs and A-BMSCs (termed Y-apoVs and A-apoVs, respectively) via differential centrifugation. The physicochemical properties (morphology, size, zeta potential, and yield) and protein markers of both apoV populations were systematically characterized. Subsequently, Y-BMSCs were divided into a proliferation medium (PM) group, osteogenic induction medium (OM) group, OM + Y-apoVs group, and OM + A-apoVs group. The osteoinductive efficacy in vitro was evaluated by alkaline phosphatase (ALP) staining, alizarin red S staining, and quantitative real-time PCR detection of key osteogenic genes, including runt-related transcription factor 2 (RUNX2), ALP, osteopontin (OPN), and osterix (OSX). Approved by the Institutional Animal Care and Use Committee, in vivo biodistribution (labelling apoVs with the red fluorescent dye PKH26) and bone regeneration efficacy were assessed in 18-month-old osteoporotic C57BL/6 mice. Mice were randomly divided into a control group (injected with phosphate-buffered saline), Y-apoVs group (injected with Y-apoVs), and A-apoVs group (injected with A-apoVs). Following tail-vein injection administration of apoVs for 8 weeks, bone regeneration was evaluated via micro-computed tomography and histological analysis. Results: Electron microscopy and particle size analysis revealed that both Y-apoVs and A-apoVs displayed typical biconcave discoid structures, with diameters mainly ranging from 100 to 500 nm. Western blot assays confirmed high expression of universal vesicle markers (CD9, CD63, CD81) and the apoptotic marker Fas in both groups. Y-apoVs and A-apoVs exhibited indistinguishable morphologies, size distributions, zeta potentials, and yields. In vitro experiments showed that, compared with the PM group, the OM, OM + A-apoVs, and OM + Y-apoVs groups significantly enhanced ALP activity, calcium nodule formation, and the expression of osteogenic genes (RUNX2, ALP, OPN, OSX) in recipient Y-BMSCs (P<0.05); moreover, compared with the OM and OM + A-apoVs groups, the OM + Y-apoVs group exhibited a more significant promoting effect (P<0.05). An in vivo analysis demonstrated that tail-vein-injected apoVs effectively homed to bone tissue. Moreover, the Y-apoVs group significantly improved trabecular microarchitecture, bone mineral density, and bone volume fraction in aged mice, exhibiting superior therapeutic efficacy over the control and A-apoVs groups (P<0.05) Conclusion The senescence state of parental cells significantly impairs the osteoinductive ability of BMSC-derived apoVs. Y-apoVs are effective biological agents for the treatment of age-related bone loss.

OBJECTIVE: To evaluate the protective effects of gentiopicroside (GPS) against reactive oxygen species (ROS)-induced NOD-like receptor family, pyrin domain containing 3 (NLRP3) inflammasome activation in endothelial cells, aiming to reduce atherosclerosis. METHODS: Eight-week-old male ApoE-deficient mice were randomly divided into 2 groups (n=10 per group): the vehicle group and the GPS treatment group. Both groups were fed a high-fat diet for 16 weeks. GPS (40 mg/kg per day) was administered by oral gavage to the GPS group, while the vehicle group received an equivalent volume of the vehicle solution. At the end of the treatment, blood and aortic tissues were collected for assessments of atherosclerosis, lipid profiles, oxidative stress, and molecular expressions related to NLRP3 inflammasome activation, ROS production, and apoptosis. Additionally, in vitro experiments on human aortic endothelial cells treated with oxidized low-density lipoprotein (ox-LDL) were conducted to evaluate the effects of GPS on NLRP3 inflammasome activation, pyroptosis, apoptosis, and ROS production, specifically examining the role of the sirtuin 1 (SIRT1)/nuclear factor erythroid 2-related factor 2 (Nrf2) pathway. SIRT1 and Nrf2 inhibitors were used to confirm the pathway's role. RESULTS: GPS treatment significantly reduced atherosclerotic lesions in the en face aorta (P<0.01), as well as in the thoracic and abdominal aortic regions, and markedly decreased sinus lesions within the aortic root (P<0.05 or P<0.01). Additionally, GPS reduced oxidative stress markers and proinflammatory cytokines, including interleukin (IL)-1 β and IL-18, in lesion areas (P<0.05, P<0.01). In vitro, GPS inhibited ox-LDL-induced NLRP3 activation, as evidenced by reduced NLRP3 (P<0.01), apoptosis-associated speck-like protein containing a CARD, cleaved-caspase-1, and cleaved-gasdermin D expressions (all P<0.01). GPS also decreased ROS production, apoptosis, and pyroptosis, with the beneficial effects being significantly reversed by SIRT1 or Nrf2 inhibitors. CONCLUSION GPS exerts an antiatherogenic effect by inhibiting ROS-dependent NLRP3 inflammasome activation via the SIRT1/Nrf2 pathway.
NLR Family, Pyrin Domain-Containing 3 Protein/metabolism* ; Reactive Oxygen Species/metabolism* ; Iridoid Glucosides/therapeutic use* ; NF-E2-Related Factor 2/metabolism* ; Animals ; Atherosclerosis/metabolism* ; Inflammasomes/drug effects* ; Male ; Sirtuin 1/metabolism* ; Signal Transduction/drug effects* ; Humans ; Endothelial Cells/pathology* ; Mice ; Oxidative Stress/drug effects* ; Apoptosis/drug effects* ; Lipoproteins, LDL ; Mice, Inbred C57BL

An assay was established for detection of toxigenic Clostridioides difficile by combining microfluidic chip analysis with immunochromatography,and its performance was evaluated and compared with those of the Xpert C.difficile/Epi and VIDAS CD AB tests.Primer pairs were designed according to the tcdB and tpi genes in C.difficile.The specificity,limit of detection,reproducibility,and stability were evaluated.A total of 215 stool samples from patients with diarrhea were collected and tested in parallel with the Xpert C.difficile/Epi,VIDAS CDAB,and our assay.C.difficile was isolated from samples,and the tcdB gene was identified when discrepant results were obtained from the three above assays.Our assay showed no cross-reaction with other diarrhea-associated pathogens.Its reproducibility was 100％in testing of two standard plasmids containing tcdB and tpi genes at two concentrations(105 and 102 copies/μL).Two standard plasmids were detected after the PCR and immunochromatography reagents had been stored for 3,6,9,and 12 months,and all the results were posi-tive.The limit of detection was 10 copies/μL for toxigenic C.difficile.Testing of 33 samples positive for C.difficile with our assay(33/215,15.3％)yielded findings statistically coherent with those of the Xpert C.difficile/Epi test(kappa value=0.965).The sensitivity,specificity,positive predictive value,and negative predictive value of our assay,with respect to Xpert C.difficile/Epi as the standard,were 94.3％,100.0％,100.0％,and 98.9％;these values were significantly higher than those of VIDAS CDAB(60.0％,98.9％,91.3％,and 92.7％)(Kappa=0.714,OR=157.50,95％CI:62.03-847.28,P=0.013).In conclusion,our newly developed assay is specific,stable,and reproducible,and may be used for rapid and accu-rate detection of toxigenic C.difficile.The assay could be used for C.difficile infection screening in outpatient and emergen-cy,community medical service center,and epidemiological settings.

Objective:To compare the prognostic performance of six trauma scoring systems—Injury Severity Score (ISS), Acute Physiology and Chronic Health EvaluationⅡ (APACHE Ⅱ), Sequential Organ Failure Assessment (SOFA), Prehospital Index (PHI), Revised Trauma Score (RTS), and the Circulation, Respiration, Abdomen, Motor, Speech (CRAMS) score—in predicting 28-day mortality among patients with severe trauma in Southwest China.Methods:A retrospective cohort study was conducted involving 479 patients with severe trauma admitted to the First Affiliated Hospital of Army Medical University between January 2018 and October 2023. Inclusion criteria were: ① age ≥16 years; ② Injury Severity Score (ISS) ≥16; ③ admission within 24 hours post-injury. Exclusion criteria included: ① severe underlying chronic conditions; ② burns or electrical injuries; ③ incomplete clinical data. Based on 28-day outcomes, patients were stratified into a survival group ( n=424) and a death group ( n=55). All patients received standardized resuscitation and damage control interventions. Prehospital scores (PHI, RTS, CRAMS) were recorded at admission. In-hospital scores (ISS, APACHE Ⅱ, SOFA) were calculated using the worst physiological parameters within the first 24 hours. The Mann-Whitney U test was used for group comparisons. Discriminative ability was assessed by the area under the receiver operating characteristic curve (AUC), with pairwise comparisons using DeLong's test. Decision curve analysis (DCA) was performed to evaluate net clinical benefit. Results:The RTS and CRAMS scores were significantly higher in the survival group, whereas ISS, APACHE Ⅱ, SOFA, and PHI scores were significantly lower in the death group (all P<0.05). The AUC values for predicting 28-day mortality, in descending order, were: APACHE Ⅱ (0.917), RTS (0.897), SOFA (0.873), PHI (0.848), CRAMS (0.831), and ISS (0.708). No significant difference in AUC was found between APACHE Ⅱ and RTS ( P=0.325). DCA showed that across most decision thresholds, both APACHE Ⅱ and RTS provided greater net clinical benefit than "treat-all" or "treat-none" strategies and other scores. Conclusions:Among the six scoring systems, APACHE Ⅱ demonstrated the highest predictive accuracy for 28-day mortality in severe trauma patients, though its efficacy was comparable to RTS. DCA confirmed their superior clinical utility. A two-phase assessment strategy—using prehospital RTS for rapid triage followed by in-hospital APACHE Ⅱ for dynamic monitoring—is recommended to optimize clinical decision-making and improve patient outcomes in Southwest China.

Objective To assess the clinical value of a novel surgical technique—Tubeless subxiphoid uniportal video-assisted thoracoscopic surgery with percutaneous suspension technique via balance-shaped sternal elevation device in the resection of anterior mediastinal masses. Methods Patients who underwent tubeless subxiphoid uniportal video-assisted thoracoscopic surgery via balance-shaped sternal elevation device in anterior mediastinal masses process at the Department of Thoracic Surgery, West China Hospital, Sichuan University from March to April 2025 were included, and their clinical data were analyzed. Results A total of 4 patients were included, with 2 males and 2 females, aged 58-75 years. The diameter of the tumor was 2.5-3.0 cm. The operation time was 60.0-150.0 min, intraoperative blood loss was 5-10 mL, pain score on the 3rd day after surgery was 0 points, and postoperative hospital stay was 2-3 days. All patients achieved complete resection of the masses and thymus without perioperative complications. Conclusion The tubeless subxiphoid uniportal video-assisted thoracoscopic surgery with percutaneous suspension technique via balance-shaped sternal elevation device technique optimizes surgical visualization and instrument maneuverability while avoiding complications related to conventional anesthesia and tubing, thereby markedly enhancing the minimally invasive profile of anterior mediastinal masses resections. In addition to maintaining procedural safety, this approach effectively reduces postoperative pain and accelerates patient recovery, highlighting its potential for widespread clinical adoption.

This study aimed to explore the therapeutic mechanisms of Colquhounia Root Tablets(CRT) in treating diabetic kidney disease(DKD) by integrating biomolecular network mining with animal model verification. By analyzing clinical transcriptomics data, an interaction network was constructed between candidate targets of CRT and DKD-related genes. Based on the topological eigenvalues of network nodes, 101 core network targets of CRT against DKD were identified. These targets were found to be closely related to multiple pathways associated with type 2 diabetes, immune response, and metabolic reprogramming. Given that immune-inflammatory imbalance driven by metabolic reprogramming is one of the key pathogenic mechanisms of DKD, and that many core network targets of CRT are involved in this pathological process, receptor for advanced glycation end products(RAGE)-reactive oxygen species(ROS)-phosphatidylinositol 3-kinase(PI3K)-protein kinase B(AKT)-nuclear factor-κB(NF-κB)-NOD-like receptor family pyrin domain containing 3(NLRP3) signaling axis was selected as a candidate target for in-depth research. Further, a rat model of DKD induced by a high-sugar, high-fat diet and streptozotocin was established to evaluate the pharmacological effects of CRT and verify the expression of related targets. The experimental results showed that CRT could effectively correct metabolic disturbances in DKD, restore immune-inflammatory balance, and improve renal function and its pathological changes by inhibiting the activation of the RAGE-ROS-PI3K-AKT-NF-κB-NLRP3 signaling axis. In conclusion, this study reveals that CRT alleviates the progression of DKD through dual regulation of metabolic reprogramming and immune-inflammatory responses, providing strong experimental evidence for its clinical application in DKD.
Animals ; Diabetic Nephropathies/metabolism* ; Receptor for Advanced Glycation End Products/genetics* ; NF-kappa B/genetics* ; Signal Transduction/drug effects* ; Rats ; NLR Family, Pyrin Domain-Containing 3 Protein/genetics* ; Proto-Oncogene Proteins c-akt/genetics* ; Drugs, Chinese Herbal/administration & dosage* ; Male ; Phosphatidylinositol 3-Kinases/genetics* ; Reactive Oxygen Species/metabolism* ; Humans ; Plant Roots/chemistry* ; Rats, Sprague-Dawley ; Tablets/administration & dosage*

This study aims to explore the protective effect of Chaihu Jia Longgu Muli Decoction on rats with heart failure after myocardial infarction, and to clarify its possible mechanisms, providing a new basis for basic research on the mechanism of classic Chinese medicinal formula-mediated inflammatory response in preventing and treating heart failure induced by apoptosis after myocardial infarction. A heart failure model after myocardial infarction was established in rats by coronary artery ligation. The rats were divided into sham group, model group, and low, medium, and high-dose groups of Chaihu Jia Longgu Muli Decoction, with 10 rats in each group. The low-dose, medium-dose, and high-dose groups of Chaihu Jia Longgu Muli Decoction were given 6.3, 12.6, and 25.2 g·kg~(-1) doses by gavage, respectively. The sham group and model group were given an equal volume of distilled water by gavage once daily for four consecutive weeks. Cardiac function was assessed using color Doppler echocardiography. Myocardial pathology was detected by hematoxylin-eosin(HE) staining, apoptosis was measured by TUNEL assay, and mitophagy was observed by transmission electron microscopy. The levels of tumor necrosis factor-α(TNF-α), interleukin(IL)-1β, and N-terminal pro-B-type natriuretic peptide(NT-proBNP) in serum were detected by enzyme-linked immunosorbent assay(ELISA). The expression of apoptosis-related proteins B-cell lymphoma 2(Bcl-2), Bcl-2-associated X protein(Bax), and cleaved caspase-3 was detected by Western blot. Additionally, the expression of phosphorylated nuclear transcription factor-κB(NF-κB) p65(p-NF-κB p65)(upstream) and nuclear factor kappa B inhibitor alpha(IκBα)(downstream) in the NF-κB signaling pathway was assessed by Western blot. The results showed that compared with the sham group, left ventricular ejection fraction(LVEF) and left ventricular short axis shortening(LVFS) in the model group were significantly reduced, while left ventricular end diastolic diameter(LVEDD) and left ventricular end systolic diameter(LVESD) increased significantly. Myocardial tissue damage was severe, with widened intercellular spaces and disorganized cell arrangement. The apoptosis rate was increased, and mitochondria were enlarged with increased vacuoles. Levels of TNF-α, IL-1β, and NT-proBNP were elevated, indicating an obvious inflammatory response. The expression of pro-apoptotic factors Bax and cleaved caspase-3 increased, while the anti-apoptotic factor Bcl-2 decreased. The expression of p-NF-κB p65 was upregulated, and the expression of IκBα was downregulated. In contrast, the Chaihu Jia Longgu Muli Decoction groups showed significantly improved of LVEF, LVFS and decreased LVEDD, LVESD compared to the model group. Myocardial tissue damage was alleviated, and intercellular spaces were reduced. The apoptosis rate decreased, mitochondrial volume decreased, and the levels of TNF-α, IL-1β, and NT-proBNP were lower. The expression of pro-apoptotic factors Bax and cleaved caspase-3 decreased, while the expression of the anti-apoptotic factor Bcl-2 increased. Additionally, the expression of p-NF-κB p65 decreased, while IκBα expression increased. In summary, this experimental study shows that Chaihu Jia Longgu Muli Decoction can reduce the inflammatory response and apoptosis rate in rats with heart failure after myocardial infarction, which may be related to the regulation of the IκBα/NF-κB signaling pathway.
Animals ; Apoptosis/drug effects* ; Drugs, Chinese Herbal/administration & dosage* ; Rats ; Myocardial Infarction/physiopathology* ; Male ; NF-kappa B/genetics* ; Heart Failure/etiology* ; Rats, Sprague-Dawley ; Signal Transduction/drug effects* ; NF-KappaB Inhibitor alpha/genetics* ; Humans ; Tumor Necrosis Factor-alpha/genetics*

This study aims to explore the intervention effects of isorhamnetin(Isor) on acute lung injury(ALI) and its regulatory effects on pyroptosis mediated by the NOD-like receptor family pyrin domain containing 3(NLRP3)/apoptosis-associated speck-like protein containing a CARD(ASC)/cysteine aspartate-specific protease-1(caspase-1) axis. In the in vivo experiments, 60 BALB/c mice were divided into five groups. Except for the control group, the other groups were administered Isor by gavage 1 hour before intratracheal instillation of LPS to induce ALI, and tissues were collected after 12 hours. In the in vitro experiments, RAW264.7 cells were divided into five groups. Except for the control group, the other groups were pretreated with Isor for 2 hours before LPS stimulation and subsequent assessments. Hematoxylin-eosin(HE) staining was used to observe pathological changes in lung tissue, while lung swelling, protein levels in bronchoalveolar lavage fluid(BALF), and myeloperoxidase(MPO) levels in lung tissue were measured. Cell proliferation toxicity and viability were assessed using the cell counting kit-8(CCK-8) method. Enzyme-linked immunosorbent assay(ELISA) was used to detect the levels of interleukin-1β(IL-1β), IL-6, IL-18, and tumor necrosis factor-α(TNF-α). Protein levels of NLRP3, ASC, cleaved caspase-1, and the N-terminal fragment of gasdermin D(GSDMD-N) were evaluated using immunohistochemistry, immunofluorescence, and Western blot. The results showed that in the in vivo experiments, Isor significantly improved pathological damage in lung tissue, reduced lung swelling, protein levels in BALF, MPO levels in lung tissue, and levels of inflammatory cytokines such as IL-1β, IL-6, IL-18, and TNF-α, and inhibited the high expression of the NLRP3/ASC/caspase-1 axis and the pyroptosis core gene GSDMD-N. In the in vitro experiments, the safe dose of Isor was determined through cell proliferation toxicity assays. Isor reduced cell death and inhibited the expression levels of the NLRP3/ASC/caspase-1 axis, GSDMD-N, and inflammatory cytokines. In conclusion, Isor may alleviate ALI by modulating pyroptosis mediated by the NLRP3/ASC/caspase-1 axis.
Animals ; Pyroptosis/drug effects* ; NLR Family, Pyrin Domain-Containing 3 Protein/genetics* ; Acute Lung Injury/physiopathology* ; Mice ; Mice, Inbred BALB C ; Quercetin/pharmacology* ; Caspase 1/genetics* ; CARD Signaling Adaptor Proteins/genetics* ; Male ; RAW 264.7 Cells ; Humans ; Lung/metabolism*