Osteoarthritis (OA) is a prevalent degenerative joint disease characterized by progressive cartilage destruction, yet the heterogeneity of transitional chondrocyte states and their contributions to pathogenesis remain incompletely understood. In this study, we constructed a single-cell transcriptomic atlas of human knee cartilage from 16 OA samples and 2 non-OA normal donors. High-resolution sub-clustering of the prehypertrophic chondrocytes (preHTC) compartment revealed eight transcriptionally distinct subtypes, among which CRIP1+ preHTC emerged as the most significantly expanded population in OA, particularly in weight-bearing regions. Pseudotemporal trajectory inference and regulon analysis identified EGR3 as a candidate key regulator for CRIP1+ preHTC. High-dimensional weighted gene coexpression network analysis (hdWGCNA) demonstrated matrix-remodeling and proteostasis-stress transcriptional programs were highly active in CRIP1+ preHTC. Cell-cell communication analysis uncovered that CRIP1+ preHTC acts as a central signaling hub in OA, with markedly enhanced FN1 signaling pathway. Spatial transcriptomics confirmed that CRIP1+ preHTC co-localizes with prefibroblasts (preFC) in pathological niches and interacts with other cells in the OA microenvironment. Finally, a machine-learning-based 10-gene classifier (JUN, MCOLN3, RHOC, AKR7A2, HSPG2, AK4, B4GALT2, CLSTN1, LRRC41, and GADD45A) derived from CRIP1+ preHTC-associated hub genes accurately discriminated OA from normal tissues in two independent bulk RNA-seq cohorts. Collectively, these findings suggest that CRIP1+ preHTC may represent a pathogenic cell state in OA, provide a multi-layered molecular framework that links a specific chondrocyte transitional subset to cartilage degradation, and offer a potentially useful gene signature for OA diagnosis.

Alcohol use is a major coexposure in patients taking acetaminophen (APAP), yet how subchronic alcohol exposure reshapes hepatic vulnerability to APAP remains incompletely defined. In this study, we examined the effects of chronic alcohol intake on APAP-induced liver injury in mice. Our results indicated that subchronic alcohol exposure significantly increased the mortality rate in APAP-overdosed mice. Alcohol pretreatment also exacerbated APAP-induced hepatic damage, as evidenced by elevated serum ALT and AST levels, enhanced inflammatory responses, and aggravated oxidative stress. Quantitative polymerase chain reaction (qPCR) analysis revealed that alcohol consumption suppressed mitochondrial biogenesis (MB)-related genes, including Pgc-1α and transcription factor A, mitochondrial (TFAM). Notably, macrophage depletion via clodronate liposomes not only attenuated hepatocyte injury and inflammation induction but also restored the expression of MB/function-related genes, including Pgc-1α, TFAM, and Opa1. These findings suggest macrophage-involved inflammatory amplification may aggravate APAP-induced liver injury, at least in part, by impairing mitochondrial adaptive repair responses.

Liver cirrhosis represents the irreversible end-stage of various chronic liver diseases, whereas hepatic fibrosis, characterized by hepatic stellate cell (HSC) activation, constitutes the critical pathological intermediate in this progression. In this study, we investigated the antifibrotic effects of loganin, a natural iridoid glycoside, and elucidated its underlying mechanisms in bile duct ligation (BDL)-induced cholestatic liver injury and hepatic fibrosis and in TGF-β1-stimulated HSCs. Sprague-Dawley rats subjected to BDL surgery received loganin for 14 days, resulting in marked amelioration of hepatomegaly, inflammatory infiltration, hepatocyte necrosis, and collagen deposition, alongside restoration of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), and total bilirubin (T-BIL) levels and suppression of fibrogenic gene expression (collagen type I alpha 1 chain [COL1α1], alpha-smooth muscle actin [α-SMA], and fibronectin). Moreover, loganin attenuated hepatic autophagy, as evidenced by reduced Beclin-1 expression and LC3B-II/I ratio, with decreased colocalization of α-SMA and LC3B. Bioinformatics analysis and molecular docking indicated that epidermal growth factor receptor (EGFR) may be the target protein through which loganin exerts its antifibrotic effect. Experimental validation demonstrated that loganin suppressed BDL-induced EGFR phosphorylation in vivo. In TGF-β1-activated HSC-T6 cells, loganin inhibited cell activation, downregulated fibrosis markers, impaired autophagic flux, and suppressed TGF-β1-induced upregulation of Beclin-1 and LC3B-II/I. Critically, cotreatment with the EGFR agonist EGF reversed the autophagy-inhibitory and antifibrotic effects of loganin, whereas the EGFR inhibitor AG-1478 mimicked loganin's actions, confirming EGFR signaling as the functional mediator. Overall, these results indicated that loganin could alleviate liver injury and hepatic fibrosis in BDL rats by inhibiting autophagy and activation of HSCs, and this protective effect is mediated by the EGFR pathway.

Activation of the NLRP3 inflammasome and its downstream cytokines IL-1β and IL-18 has been increasingly associated with the inflammatory mechanisms underlying diabetic nephropathy (DN), the leading cause of chronic kidney disease and end-stage renal disease worldwide. This study investigated the expression of these inflammatory mediators in renal tissue from patients with DN. Eighty renal biopsies from adult patients (≥18 years) with a histopathological diagnosis of DN and 22 control samples obtained from autopsies were analyzed. NLRP3, IL-1β, and IL-18 expression was assessed by immunohistochemistry, and semi-quantitative analysis was performed to determine the proportion of immunostained cells in the glomerular and interstitial compartments. A significant increase in immunostaining for all three mediators was observed in mesangial cells, podocytes, endothelial cells, and the interstitium of DN samples compared with controls (p < 0.05). Positive and significant correlations were found between NLRP3 and IL-1β in endothelial cells (p < 0.0001; rS = 0.4720) and in the interstitium (p = 0.0230; rS = 0.2540), as well as between NLRP3 and IL-18 in podocytes (p = 0.0055; rS = 0.3074). Moreover, interstitial expression of NLRP3 and IL-1β correlated positively with serum creatinine levels (p = 0.0015; rS = 0.3708; and p = 0.0029; rS = 0.3480, respectively), suggesting an association between inflammasome-mediated inflammation and renal dysfunction. Collectively, these findings indicate that the NLRP3/IL-1β/IL-18 axis plays a central role in glomerular and interstitial injury in DN, supporting its potential as a prognostic biomarker and a promising therapeutic target for disease modulation.

Qingxin Tongmai Yin (QXTMY), a classic traditional Chinese medicine (TCM) formula widely used for cardiovascular and cerebrovascular diseases, has unclear multitarget mechanisms against atherosclerosis (AS). This study integrated network pharmacology with experimental validation to investigate these mechanisms. Network pharmacology identified 128 bioactive compounds and 135 potential targets of QXTMY, highlighting key anti-AS targets such as TNF, IL6, insulin (INS), IL1B, MMP9, CCL2, and ALB, with the AGE-RAGE signaling pathway as a central mechanism; principal active ingredients included quercetin, kaempferol, luteolin, and cryptotanshinone. In vitro experiments using THP-1-derived macrophages induced with phorbol-12-myristate-13-acetate (PMA) and oxidized low-density lipoprotein (ox-LDL) showed that QXTMY significantly suppressed inflammatory responses, inhibited foam cell formation, and downregulated the AGE-RAGE signaling pathway, as measured by Western blot, ELISA, cholesterol quantification, and Nile Red staining. These findings suggest that QXTMY may exert protective effects against AS, potentially via suppression of the AGE-RAGE-mediated inflammatory axis. However, in vivo validation is still required to support these preliminary conclusions.

Prolonged air leak (PAL) remains one of the most frequent and clinically consequential complications after lung resection, prolonging chest tube duration, delaying recovery, increasing postoperative morbidity, and expanding healthcare resource utilization. Although traditionally approached as a mechanical failure of alveolar-pleural sealing or surgical technique, PAL is increasingly understood as a heterogeneous postoperative syndrome in which persistent air leak dynamics intersect with impaired inflammatory and reparative biology. Surgical manipulation, one-lung ventilation, parenchymal injury, and patient-specific vulnerability may activate cytokine signaling, epithelial barrier disruption, oxidative stress, extracellular matrix remodeling, glycocalyx degradation, and delayed wound repair. This narrative review synthesizes clinical, translational, and experimental evidence on the pathogenesis of PAL, with particular emphasis on inflammatory mediators and biomarker-defined mechanisms relevant to failed closure of alveolar-pleural fistulas (APFs). Candidate biomarkers are discussed according to the biological domains they reflect, including systemic inflammatory burden and poor healing reserve (C-reactive protein [CRP], IL-6, and serum albumin), epithelial injury and damage-associated signaling (high-mobility group box 1 [HMGB1]/receptor for advanced glycation end products [RAGE] and sRAGE), oxidative epithelial stress (4-hydroxynonenal [4-HNE] and malondialdehyde [MDA]), protease-mediated matrix and junctional remodeling (matrix metalloproteinase [MMP]-9), glycocalyx and barrier disruption (syndecan [SDC]-1), and upstream inflammatory regulation (histone deacetylase 6 [HDAC6]-related pathways). The review also considers how quantitative digital chest drainage parameters may complement molecular biomarkers by capturing the physiologic expression of persistent air leak. By integrating these mechanistic and technological signals, this review proposes a conceptual framework for biomarker-informed perioperative risk stratification, PAL phenotyping, and individualized prevention and management. Because most biomarker-driven strategies remain investigational, prospective validation is required before routine clinical implementation.

The current management of acute pancreatitis (AP) primarily relies on supportive measures, such as fluid resuscitation, nutritional support, and infection control. However, these approaches do not adequately address the core drivers of the disease. This limitation arises from an incomplete understanding of the progression from local injury to systemic inflammation and repair, which is a dynamic process with underlying immunoregulatory mechanisms that remain insufficiently characterized. Macrophages are the key effector cells involved throughout the disease course and exhibit pronounced spatiotemporal heterogeneity during this progression. Therefore, they are central to decoding the evolution of the disease and facilitating precise interventions. In terms of spatial dynamics, tissue-resident macrophages (TRMs), which are derived from embryonic sources, and monocyte-derived macrophages (MDMs) recruited from the bone marrow serve functionally complementary roles. Their relative dominance shifts in conjunction with disease progression rather than remaining static. Temporally, the macrophage phenotype undergoes a programmed evolution, beginning with an early phase dominated by M1 proinflammatory responses, transitioning through an intermediate phase where injury and repair coexist, and culminating in a late phase characterized by M2-dominated reparative coordination. This evolution is accompanied by corresponding metabolic reprogramming. Recent single-cell and spatial multiomics studies have unveiled a functional continuum that goes beyond the traditional M1/M2 dichotomy, revealing a rich diversity of cellular subsets and their spatial niches. This insight shifts targeting strategies from broad anti-inflammatory interventions toward more precise modulation aimed at specific phases, subsets, and regions. This review systematically examines the design principles, strengths, and limitations of three classes of intervention-molecular targeting, bioactive natural products, and nanoscale delivery-and identifies the obstacles that continue to impede their clinical translation. Building on this analysis, we propose a dual-dimensional (spatiotemporal) strategy of precise modulation, integrating single-cell and spatial omics, chronobiological principles, and the traditional Chinese medical concept of yin shi zhi yi (adapting treatment to timing), with the aim of shifting AP therapy from symptomatic support toward cause-directed repair.

Alcohol use is a major global health issue, causing about 3.3 million deaths each year, or roughly 5.9% of all deaths worldwide. Alcohol-related liver disease develops in stages: steatosis, steatohepatitis, fibrosis, cirrhosis, and hepatocellular carcinoma. If alcohol consumption is discontinued at early stages, alcohol-related fatty liver disease can be reversed; however, continued exposure leads to progressive liver injury and increased mortality risk. Current diagnostic tools lack sufficient sensitivity and specificity to detect early-stage disease or accurately assess disease progression. This highlights the need for reliable and mechanistically relevant biomarkers for early diagnosis and staging. This review examines alterations in gut microbiota across different stages of alcohol-associated liver disease and evaluates gut microbiota-associated biomarkers, including microbial metabolites, in the context of their potential diagnostic and prognostic utility. In addition, the review discusses the limitations of existing biomarkers and highlights the emerging role of microbiome-derived signals in reflecting disease mechanisms. These findings suggest that gut microbiota-related biomarkers may provide a promising but still evolving approach for improving early detection and understanding disease progression in alcohol-associated liver disease.