Background: Hypertrophic scar (HS) arises from aberrant cutaneous wound healing, characterized by uncontrolled myofibroblast proliferation and disproportionate extracellular matrix (ECM) accumulation. Breviscapine, a natural compound, suppresses inflammation and fibrosis. This study aimed to evaluate breviscapine as a therapeutic avenue against HS and unravel its molecular basis. Methods: In vivo, a rabbit ear HS model was established and treated with different doses of breviscapine. Scar morphology was macroscopically evaluated. Fibrosis-related markers (collagen type I alpha 1 [COL1A1], collagen type III alpha 1 [COL3A1], actin alpha 1, skeletal muscle [ACTA1], alpha-smooth muscle actin [α-SMA]) were quantified by quantitative real-time PCR (qRT-PCR) and Western blotting. α-SMA expression, collagen fiber organization and transforming growth factor-β1 (TGF-β1) levels were assessed by immunofluorescence staining, Masson staining, and immunohistochemistry/Western blotting, respectively. In vitro, HS fibroblasts (HSFs) were exposed to breviscapine, and cell counting kit-8 (CCK-8) assay, Sirius red staining and Western blotting were separately performed to determine cell viability, collagen production, and the expression of fibrosis-related markers, TGF-β1, and the Akt-mTOR pathway. Results: In vivo, breviscapine treatment attenuated rabbit ear scar formation, while diminishing COL1A1/COL3A1/ACTA1 mRNA levels and collagen I/collagen III/α-SMA/TGF-β1 protein levels and improving collagen fiber density and arrangement (p < 0.001). In vitro, breviscapine inhibited HSF viability, reversed TGF-β1-stimulated collagen overproduction, and suppressed the protein expression of fibrosis-related markers (p < 0.05). Notably, breviscapine attenuated the protein expression of TGF-β1 and the Akt-mTOR pathway in HSFs (p < 0.01). Conclusion: Breviscapine exerts anti-HS effects by targeting the TGF-β1 signaling and the Akt-mTOR pathway, providing a novel natural therapeutic candidate for HS management with anti-fibrotic, anti-proliferative, and pathway-targeted effects. Further investigation is warranted to validate its efficacy in clinical settings and to explore its long-term safety and optimal delivery strategies.
Mucosal inflammatory disorders, including inflammatory bowel disease (IBD), have increased steadily in prevalence worldwide. These conditions follow a protracted clinical course with high complication rates and no curative therapies. The breakdown of intestinal mucosal immune homeostasis is central to their pathogenesis. Among the key regulatory pathways, the balance between regulatory T cells (Tregs) and T helper 17 (Th17) cells is a pivotal determinant of mucosal immune tolerance and the containment of excessive inflammatory responses. This review synthesizes current knowledge on the biological characteristics, differentiation regulatory networks, and functional crosstalk between Tregs and Th17 cells. We describe how the mutual antagonism between the lineage-specific transcription factors Forkhead box P3 (Foxp3) and retinoic acid-related orphan receptor gamma t (RORγt) acts as a core molecular switch governing this balance, while T cell receptor (TCR) signaling, the cytokine microenvironment, and epigenetic modifications form its upstream regulatory network. We further discuss the precise mechanisms by which commensal microbiota and their metabolites, intestinal epithelial barrier integrity, and regional heterogeneity of mucosal tissues modulate the Treg/Th17 balance, and we dissect how disruption of this balance drives IBD onset and progression. We also summarize Treg/Th17-related biomarkers for IBD diagnosis and disease monitoring and emerging therapeutic strategies targeting this balance. Finally, we outline priority research directions, emphasizing the translational potential of single-cell multi-omics and spatial transcriptomics for achieving tissue-specific precision regulation of Treg/Th17 balance in IBD.
Background: Immune checkpoint blockade targeting the Programmed Death-1 (PD-1)/Programmed Death-Ligand 1 (PD-L1) axis has demonstrated clinical activity in esophageal squamous cell carcinoma (ESCC). However, therapeutic responses remain limited in a substantial proportion of patients. Metabolic reprogramming and lactate accumulation are hallmarks of the tumor microenvironment and have been implicated in immune suppression, yet the epigenetic mechanisms linking lactate metabolism to immune checkpoint regulation in ESCC remain poorly understood. The aim of this study was to investigate whether lactate-induced histone lactylation regulates PD-L1 expression and contributes to tumor-intrinsic PD-L1 regulation in ESCC.Methods: ESCC cell lines were treated with exogenous lactate and metabolic modulators to evaluate PD-L1 expression and histone H3 lysine 18 lactylation (H3K18la). The involvement of the lactyltransferase p300 was assessed using pharmacological inhibition. T cell function was examined using an in vitro co-culture system with activated Jurkat T cells. In vivo validation was performed using a nude mouse xenograft model, followed by immunohistochemical analysis.Results: Lactate treatment significantly increased PD-L1 expression in ESCC cells (p < 0.001), accompanied by elevated global H3K18la (p < 0.001). Inhibition of glycolysis reduced basal H3K18la and PD-L1 expression (p < 0.001), which was restored by exogenous lactate (p < 0.001). Pharmacological inhibition of p300 abrogated lactate-induced H3K18la and PD-L1 upregulation (p < 0.001), indicating a p300-dependent mechanism. Functionally, lactate-treated ESCC cells reduced the expression of the activation-associated cytokines Interleukin-2 (IL-2) and Interferon-gamma (IFN-γ) in co-cultured Jurkat T cells (p < 0.001), indicative of attenuated T-cell activation, an effect partially reversed by PD-L1 knockdown. In vivo, lactate administration increased intratumoral H3K18la and PD-L1 expression in xenograft tumors (p < 0.001).Conclusions: Our findings identify a lactate-p300-H3K18 lactylation axis associated with transcriptional upregulation of PD-L1 and attenuation of T-cell activation in a Jurkat-based co-culture system, providing a metabolic-epigenetic framework for improving immunotherapeutic strategies.
Background: The metabolic characteristics of preterm infants in the early postnatal period are closely linked to their growth and development. Investigating the corresponding metabolic changes that occur during the early postnatal maturation process of preterm infants may help improve their clinical management. In this prospective cohort study, we aimed to characterize urinary metabolism in preterm infants during the first two weeks of life and explore its relationship with metabolic homeostasis regulation in the early postnatal period.Methods: Between June 1, 2021, and October 1, 2022, 36 preterm infants eligible for enrollment were recruited from the Children’s Hospital of Fudan University (Shanghai, China), and urine samples were collected from these 36 preterm infants on postnatal day 1 (PND1), postnatal day 7 (PND7), and postnatal day 14 (PND14), respectively. A longitudinal urinary metabolomics study of preterm infants was conducted using an Ultra-high performance liquid chromatography-MS/MS (UHPLC-MS/MS) technique to compare urinary metabolic characteristics between PND7 and PND1 during the first postnatal week, and between PND14 and PND7 during the second postnatal week.Results: Broad-targeted urinary metabolomics analysis of preterm infants during the first week of life identified 75 differentially expressed urinary metabolites, of which 57 were upregulated compared to birth, while 18 were downregulated. The most significantly upregulated metabolite was L-3-hydroxykynurenine (p = 0.003), while the most significantly downregulated was N-Acetyltyrosine (p < 0.001). The metabolic pathway most significantly enriched among these 75 differentially expressed metabolites was the Arginine biosynthesis (p = 0.001). Compared to the first week, only 29 differentially expressed urinary metabolites were identified in preterm infants during the second week after birth, with 22 metabolites upregulated and 7 downregulated. Among them, (3-hydroxyphenyl)-3-hydroxypropionic acid (HPHPA) was the most significantly upregulated (p = 0.001), while N-Acetylhistidine was the most significantly downregulated (p = 0.008). The metabolic pathways most significantly enriched among these 29 urinary metabolites were Glycolysis/Gluconeogenesis (p = 0.008). Cinnamoylglycine, cyclic-3',5'-GMP and lactate were common urinary differentially expressed metabolites in the first and second weeks after birth in preterm infants, and the Glycolysis/Gluconeogenesis pathway was the metabolic pathway significantly associated with urinary differentially expressed metabolites in the first two weeks after birth.Conclusion: This single-center study describes the general characteristics of urinary metabolomics in preterm infants during the first two weeks after birth. Glycometabolism was identified as a key factor influencing changes in urine metabolism in preterm infants during the first two weeks after birth, providing a reference for metabolic adaptation assessment and clinical management strategies in the early postnatal period. Clinical Trial Registration: ClinicalTrials.gov (NCT06018831).
Amyotrophic lateral sclerosis (ALS) is a progressive, life-limiting neurodegenerative disease characterized primarily by motor dysfunction, although cognitive and behavioral impairment occurs in a substantial subset of patients. Progressive functional decline and prognostic uncertainty may contribute to pronounced fear of disease progression (FoP). Increasing evidence suggests that FoP represents a distinct psychological response to the expected functional loss related to disease, rather than merely a secondary manifestation of ALS symptoms. To clarify the potential biological basis of ALS-related FoP, this review synthesizes current evidence regarding stress hormone pathways, autonomic nervous system (ANS) dysfunction, inflammation, and the gut-brain axis. Based on the available evidence, we propose that FoP may emerge from persistent mismatch between patients’ perceived disease-related threats and their adaptive capacity to these challenges. Multiple interacting systems may contribute to this process, including dysregulation of the hypothalamic-pituitary-adrenal (HPA) axis, autonomic imbalance, sleep and circadian rhythm disruption, peripheral and central inflammatory responses, and changes in the gut microbiota that may influence emotional regulation and physical symptoms. These interacting processes may create a reinforcing cycle: ALS-specific pathological features may enhance FoP, while the neuroendocrine mechanisms associated with FoP may further aggravate the severity and burden of the disease, thereby increasing the complexity of disease management. Future studies should prioritize the development of molecular biomarkers, as these biomarkers may provide valuable guidance for establishing ALS-specific FoP assessment tools and developing novel therapeutic strategies.
Background: The epidermal growth factor receptor (EGFR)-mutated lung adenocarcinoma (LUAD) exhibits significant stage-dependent molecular and spatial heterogeneity, which remains incompletely characterized. Elucidating stage-associated differences in gene expression, signaling pathways, and immune landscapes is critical for developing stage-stratified precision therapy for EGFR-mutated LUAD.Methods: This study integrated bulk transcriptome profiling and digital spatial profiling (DSP) to investigate pathological stage-associated differences. DSP was applied to 23 EGFR-mutated LUAD patients (18 early-stage and 5 late-stage) to elucidate spatial heterogeneity across three tissue compartments: tumor cells, immune cells, and macrophages. Public bulk RNA sequencing data from 38 patients with EGFR-mutated LUAD, including 29 early-stage (I+II) and 9 late-stage (III+IV) cases, were analyzed.Results: Bulk RNA sequencing identified 580 differentially expressed genes between early- and late-stage EGFR-mutated LUAD. Late-stage tumors exhibited enhanced proliferation, marked by elevated E2F, G2M checkpoint, and MYC activities, alongside activated cell cycle, DNA replication, and DNA repair pathways. Notable metabolic reprogramming was also observed, including upregulated serine and purine metabolism. Spatial transcriptomic analysis confirmed increased proliferative, oncogenic, metastatic, and glycolytic signatures in late-stage tumor cells, consistent with findings from bulk RNA sequencing. Pathway enrichment analysis across distinct compartments revealed six stage and compartment specific modules, with modules related to cell cycle, DNA repair, invasion, and metabolism significantly upregulated in the late-stage tumor cell segment. Moreover, late-stage tumors displayed an increase in exhausted CD8+ T cells and elevated expression of immune checkpoints (PD-L2, PVR in tumor cells; B7-H3, LAG3 in immune cells), contributing to an immunosuppressive microenvironment.Conclusions: These findings delineate stage-specific transcriptional, metabolic, and spatial immune reprogramming during the progression of EGFR-mutated LUAD. These results may provide new insights into the molecular mechanisms underlying tumor progression and identify potential targets for stage-stratified precision therapy in EGFR-mutated LUAD.
Background: Patchouli alcohol (PA), a tricyclic sesquiterpene from Pogostemonis Herba, exhibits anti-inflammatory, antioxidant and cardiovascular activities. Adriamycin (Adr) induces dose-dependent cardiotoxicity that progresses to irreversible heart failure (HF). This study investigated the mechanism of PA in rats with Adr-induced HF. Methods: Male Sprague Dawley (SD) rats (initial n = 15/group) received intraperitoneal Adr (2 mg/kg, twice weekly for 4 weeks) to induce HF. Two weeks after modeling, PA (30 mg/kg/d) or Icilin (20 mg/kg/d) was administered for 4 weeks. Cardiac function (left ventricular ejection fraction (LVEF)/left ventricular short axis shortening rate (LVFS)), serum brain natriuretic peptide (BNP), lactate dehydrogenase (LDH), creatine kinase (CK), nitric oxide (NO), nitric oxide synthase (NOS), aspartate aminotransferase (AST), myocardial transient receptor potential melastatin 8 (TRPM8) and apoptosis-related proteins (Bcl-2 associated X protein (Bax), B-cell Lymphoma 2 (Bcl-2), and cleaved Caspase-3) were assessed. In vitro, Adr-treated AC16 cells were used to confirm the anti-apoptotic role of TRPM8 through PA treatment, N-(4-tert-butylphenyl)-N′-(3-chloro-4-fluorophenyl)urea (BCTC), and TRPM8 manipulation. Results: In vivo, PA and Icilin ameliorated cardiac dysfunction in Adr-induced HF rats (p < 0.05), downregulated HF markers and myocardial injury factors (p < 0.05), and mitigated Adr-induced myocardial pathological damage. Both agents also upregulated myocardial TRPM8 expression (p < 0.001) and reversed Adr-induced abnormal apoptosis-related proteins (p < 0.05), thus inhibiting excessive cardiomyocyte apoptosis. In vitro, PA suppressed Adr-induced AC16 cardiomyocyte apoptosis by upregulating TRPM8 (p < 0.01), whereas TRPM8 knockdown or inhibition abrogated PA's protective effect (p < 0.001). Conclusion: This study identifies PA as a novel pharmacological enhancer of TRPM8, demonstrating that PA exerts significant cardioprotective effects in Adr-induced HF by targeting TRPM8 and inhibiting cardiomyocyte apoptosis. These findings support TRPM8 as a key mediator of PA's protective role and provide a promising therapeutic strategy for HF management.
Background: Leptomeningeal carcinomatosis (LC) remains difficult to diagnose at an early stage because current diagnostic approaches rely primarily on magnetic resonance imaging and cerebrospinal fluid (CSF) cytology, which may lack sensitivity or require repeated testing. This study aimed to develop a water-stable, biocompatible, and tumor-targeted fluorescent nanoparticle for the preliminary diagnosis of LC.Methods: CsSn0.5Pb0.5Br3@CS-FA nanoparticles were synthesized using a water-triggered strategy and characterized by X-ray diffraction, Fourier-transform infrared spectroscopy, transmission electron microscopy, photoluminescence spectroscopy, ultraviolet-visible spectroscopy, and in vivo imaging. Their water stability, in vitro targeting ability, cytocompatibility, in vivo tumor-targeting capacity, and biosafety were evaluated using Lewis lung carcinoma (LLC) cells, C2C12 cells, C57BL/6JNifdc mouse models, and clinical CSF samples. CSF samples from 7 patients with LC and 7 non-LC controls were incubated with the nanoparticles, and fluorescence intensity was detected by flow cytometry.Results: CsSn0.5Pb0.5Br3@CS-FA nanoparticles showed a stable core-shell structure and maintained fluorescence in aqueous solution for up to 329 days. In vitro imaging demonstrated stronger fluorescence in LLC cells than in C2C12 cells, and CsSn0.5Pb0.5Br3@CS-FA produced significantly higher fluorescence than non-targeted CsSn0.5Pb0.5Br3@CS nanoparticles (p < 0.001). Cell viability remained above 90% after treatment with 200 μg/mL CsSn0.5Pb0.5Br3@CS-FA. In tumor-bearing mice, fluorescence signals were detectable after injection, peaked at 24 h, and were no longer detectable at 216 h. Hematological, hepatic, renal, histological, and body-weight assessments showed no obvious biosafety concerns. In clinical CSF samples, the fluorescence intensity was markedly higher in the LC group than in the non-LC group (47.08% ± 13.56% vs. 2.75% ± 1.12%, p < 0.001).Conclusion: CsSn0.5Pb0.5Br3@CS-FA nanoparticles exhibited favorable water stability, tumor-targeting ability, fluorescence performance, and preliminary biosafety. Their enhanced fluorescence signal in LC-derived CSF samples suggests potential value as an auxiliary fluorescent tracer for LC diagnosis, although further validation in larger clinical studies is required.
Background: Automated Breast Ultrasound (ABUS) is primarily used for standardized volumetric breast imaging but has been widely considered inadequate for axillary assessment due to technical constraints. This study aims to quantitatively evaluate this assumption by comparing the axillary lymph node (ALN) detection performance of ABUS with that of Mammography (MG), Digital Breast Tomosynthesis (DBT), and Magnetic Resonance Imaging (MRI).Methods: This retrospective, single-center study screened 339 women undergoing imaging between May 2023 and December 2025; after excluding 35 patients (imaging interval >14 days or insufficient data), 304 valid axillary evaluations were included in the analysis. Patients were categorized into three cohorts based on paired imaging of the same axilla within a two-week interval: MG-DBT-ABUS (n = 304), ABUS-MRI (n = 51), and a comprehensive MG-DBT-ABUS-MRI subset (n = 16). A specialized ABUS axilla protocol involving arm hyper-abduction was utilized. ALN counts were compared using paired samples t-tests and Wilcoxon Signed-Rank tests.Results: In the MG-DBT-ABUS group, ABUS detected significantly more lymph nodes (mean 4.52 ± 1.80; median 4.00) compared to the median values of 0.00 (interquartile range (IQR): 0.0–2.0) for both MG and DBT (p < 0.001). In the ABUS-MRI group, MRI detected significantly more nodes (6.51 ± 2.70) than ABUS (4.41 ± 1.94) (p < 0.001). However, ABUS visualized approximately 68% of the lymph nodes detected by MRI. In the four-modality group (n = 16), non-parametric testing of median detection hierarchy demonstrated that MRI [6.00 (IQR: 4.0–8.0)] and ABUS [5.00 (IQR: 4.0–5.5)] were significantly superior to DBT [0.00 (IQR: 0.0–2.0)] and MG [0.00 (IQR: 0.0–1.0)]; no direct statistical comparison between DBT and MG was performed in this subset.Conclusion: ABUS is not a blind spot for the axilla. While MRI remains the current imaging reference standard, ABUS demonstrates superior quantitative detection performance compared to mammography and tomosynthesis. When performed with a dedicated protocol, ABUS serves as a valuable, non-ionizing supplementary tool for axillary lymph node assessment. While this study primarily evaluates quantitative detection performance, pathological correlation and morphological characterization warrant further investigation in future studies.
Background: Polycystic ovary syndrome (PCOS) is a common hormone disorder in women of reproductive age. It is often associated with changes in adipokine signaling, obesity, and insulin resistance. The leptin gene produces a hormone that helps control energy balance and reproductive health. The –2548G>A promoter polymorphism in this gene might affect how leptin is expressed and metabolized, but findings have differed between populations. This study aims to investigate the distribution of –2548G>A genotypes and serum leptin levels in women with PCOS.Methods: The study included 100 women with PCOS and 100 healthy controls of similar age, recruited between July 2023 and September 2025. Researchers measured physical, hormonal, and metabolic factors. They used enzyme-linked immunosorbent assay (ELISA) to check serum leptin levels and polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) to identify –2548G>A genotypes. Data analysis was done with SPSS version 25.0.Results: There were no significant differences between the groups in age, body mass index, or other biochemical and hormonal measures (p > 0.05). However, the A/A genotype of the –2548G>A polymorphism was more common in the PCOS group than in controls (p = 0.03). There was no significant link between –2548G>A genotypes and serum leptin levels.Conclusions: The –2548G>A polymorphism may raise the risk of developing PCOS, but it does not seem to affect serum leptin levels. Larger studies that include leptin receptor variants and functional tests are needed to better understand how leptin is regulated in PCOS.
Background: Traumatic optic neuropathy (TON) causes progressive retinal ganglion cell (RGC) loss and oxidative injury, leading to visual dysfunction. Ephrin B2 (EFNB2) is elevated in optic nerve injury; however, it is unclear how the fat mass and obesity-associated gene (FTO) regulates it upstream. This study examined whether RGCs are shielded from hydrogen peroxide (H2O2)-induced oxidative damage through FTO-mediated EFNB2 overexpression.Methods: Primary RGCs were isolated and exposed to H2O2 to establish an in vitro oxidative stress injury model. We first verified the concentration-dependent induction of FTO and EFNB2 by H2O2. Gain- and loss-of-function assays were performed using FTO overexpression and knockdown plasmids to assess its effects on RGC viability, apoptosis, reactive oxygen species (ROS) accumulation, caspase-3 activity, and the expression of EFNB2, erythropoietin-producing hepatoma receptor B4 (EPHB4) and apoptosis-related proteins. Rescue experiments with EFNB2 knockdown were further conducted to confirm whether EFNB2 acts as a downstream effector of FTO.Results: H2O2 treatment elevated FTO and EFNB2 expression in RGCs in a concentration-dependent manner (p < 0.01). FTO overexpression alleviated H2O2-induced reductions in cell viability, as well as increases in apoptosis, caspase-3 activation, and ROS levels (p < 0.001), while FTO knockdown exacerbated these injuries (p < 0.001). Moreover, FTO overexpression enhanced EFNB2 and EPHB4 expression, and suppressed Bax and cleaved caspase-3 levels (p < 0.001). Knockdown of EFNB2 reversed the protective effects of FTO overexpression on RGC survival, oxidative stress, and apoptosis (p < 0.001).Conclusions: FTO upregulates EFNB2 and EPHB4 expression, reduces ROS and apoptosis, and is associated with the protection of RGCs against H2O2-induced oxidative damage. The FTO/EFNB2 axis may represent a protective mechanism against oxidative damage in RGCs in vitro, warranting further investigation in traumatic optic neuropathy models.
Atypical lobular endocervical glandular hyperplasia (ALEGH) is a cervical precancerous lesion that develops independently of high-risk human papillomavirus (HPV) infection. It is regarded as a key intermediate stage in the progression from lobular endocervical glandular hyperplasia to gastric-type endocervical adenocarcinoma (G-EAC). Because these lesions are often located in the upper endocervical canal or deep cervical stroma, and their cytological and histological atypia may be subtle, ALEGH is prone to being missed or misdiagnosed in clinical practice. Accurate identification of ALEGH and assessment of its malignant potential are central to improving clinical management. In recent years, advances in multimodal imaging, artificial intelligence (AI)-assisted image analysis, preoperative targeted biopsy, and molecular pathology have provided new evidence for improving preoperative detection and risk stratification. In this review, we summarize the current evidence regarding the imaging diagnosis, preoperative pathological confirmation, molecular mechanisms, and stratified clinical management of ALEGH, with the aim of providing practical reference for early recognition, malignant risk assessment, fertility-preserving decision-making, and surgical management of high-risk ALEGH cases.
Background: Type 2 diabetes mellitus (T2DM) is one of the most common chronic metabolic diseases worldwide, and insulin resistance is the core pathophysiological basis for its development. This study aims to explore the relationship between various metabolic-related indicators and insulin resistance in patients with T2DM, and to analyze the correlation between obesity indicators, glucose and lipid metabolism indicators, and serum uric acid with insulin resistance, providing a reference for clinical identification of insulin resistance.Methods: This was a single-center retrospective study, consecutively enrolling 263 patients with type 2 diabetes who received treatment at our center between August 2022 and August 2025. Patients were divided into a non-insulin-resistant group (n = 211) and an insulin-resistant group (n = 52) based on the Homeostasis Model Assessment of Insulin Resistance (HOMA-IR). Baseline clinical data and laboratory parameters were collected, including body mass index, waist-to-hip ratio, fasting blood glucose, 2-hour postprandial blood glucose, lipid profile, glycated hemoglobin, fasting C-peptide, and serum uric acid. The triglyceride-glucose (TyG) index was also calculated. Logistic regression analysis was performed to screen for metabolic indicators associated with insulin resistance, and receiver operating characteristic (ROC) curve analysis was then conducted to evaluate the discriminative ability of these indicators for insulin resistance.Results: Patients in the insulin resistance group had significantly higher levels of body mass index (BMI), waist-to-hip ratio (WHR), triglycerides, fasting blood glucose, TyG index, fasting C-peptide, glycated hemoglobin (HbA1c), systolic blood pressure, diastolic blood pressure, and serum uric acid than the non-insulin resistance group (all p < 0.05), and a higher proportion of patients with hypertension (65.38% vs. 46.45%, p = 0.014). Multivariate logistic regression analysis showed that WHR, TyG index, fasting C-peptide, and serum uric acid were independently associated with insulin resistance. The area under the curve (AUC) for assessing insulin resistance using the combined indicators was 0.81 (95% CI: 0.74–0.88), with a sensitivity of 79.0% and a specificity of 71.0%.Conclusion: In patients with type 2 diabetes, WHR and serum uric acid levels are significantly correlated with insulin resistance status. These indicators reflect various metabolic characteristics from different perspectives, including obesity, glucose and lipid metabolism, and pancreatic β-cell function, and may provide preliminary insights into the metabolic abnormalities associated with insulin resistance in patients with type 2 diabetes.
Background: Patients with compensated early-stage liver fibrosis (F1–F2) are at risk of progressing to cirrhosis, yet routine surveillance with liver biopsy or transient elastography is limited by invasiveness, cost, or accessibility. Simple, non-invasive tools are therefore needed for risk stratification. This study aims to develop a simple risk scoring system based on conventional ultrasound features and evaluate its ability to predict progression to cirrhosis within 3 years in patients with compensated early-stage liver fibrosis (F1–F2).Methods: This retrospective study included 502 patients with biopsy-proven compensated early-stage liver fibrosis. Four ultrasound features—liver surface, liver parenchymal echogenicity, liver margin morphology, and splenic volume (calculated as length × thickness × width × 0.52)—were semiquantitatively scored (0–2 points each) at baseline. The total score (0–8) was used to classify patients into low-risk (≤4) and high-risk (>4) groups. The primary endpoint was progression to compensated cirrhosis within 3 years. A Cox proportional hazards model was used to assess the association between the risk score and cirrhosis progression. Model discrimination was evaluated using the C-index and time-dependent ROC curves. Progression-free survival curves were plotted using the Kaplan–Meier method and compared by the log-rank test.Results: Among 502 patients, 374 (74.5%) were classified as low-risk (Risk Score ≤4) and 128 (25.5%) as high-risk (Risk Score >4). The 3-year cumulative cirrhosis progression rates were 8.6% and 35.2%, respectively (log-rank p < 0.0001). The risk score was an independent predictor (hazard ratio (HR) = 1.43, 95% confidence interval (CI) 1.29–1.59, p < 0.001), with a C-index of 0.716 and a 36-month time-dependent area under the receiver operating characteristic curve (AUC) of 0.734. Bootstrap internal validation yielded an optimism-corrected C-index of 0.716. The model showed good calibration (Brier score = 0.118). Decision curve analysis demonstrated positive net benefit of the score across threshold probabilities of 0.05–0.35. The score showed higher C-index (0.716 vs. 0.650, 0.631, and 0.660) and 36-month AUC (0.734 vs. 0.679, 0.672, and 0.675) compared to FIB-4, APRI, and LSM. Adding the score to liver stiffness measurement (LSM) significantly improved reclassification (net reclassification improvement (NRI) = 0.184, p = 0.007; integrated discrimination improvement (IDI) = 0.048, p = 0.025). Albumin was also an independent protective factor (HR = 0.93, 95% CI 0.87–0.99, p = 0.024) in the multivariable sensitivity model.Conclusion: This simple risk score (0–8) based on conventional ultrasound features (liver surface, parenchymal echogenicity, liver margin, and splenic volume) is an independent predictor of progression to cirrhosis within 3 years in patients with compensated early-stage liver fibrosis. This simple scoring system may provide a convenient and cost-effective non-invasive tool for risk stratification in clinical practice; however, these findings require validation in prospective, multicenter cohorts.
Background: The intestinal microbial ecosystem has emerged as an important contributor to chronic obstructive pulmonary disease (COPD) pathogenesis. Azithromycin (AZI), a commonly used therapeutic agent for COPD, has antibacterial and anti-inflammatory effects. This study aims to explore the mechanism by which AZI regulates the intestinal microbial ecosystem to ameliorate COPD.Methods: A rat COPD model was established by cigarette smoke (CS) exposure. The animals were pretreated with AZI (0.5, 5, 50 mg/kg) with or without the adenosine monophosphate-activated protein kinase (AMPK) inhibitor Compound C. Pulmonary dysfunction and colon injury in COPD rats were evaluated by detecting forced expiratory volume within 0.3 seconds (FEV0.3), forced vital capacity (FVC), as well as hematoxylin and eosin staining. Flow cytometry was used to detect T helper 17 (Th17)/regulatory T (Treg) cell balance in colon tissue. Inflammatory factors and secretory IgA (sIgA) levels were detected using quantitative real-time polymerase chain reaction (qRT-PCR) and enzyme-linked immunosorbent assay (ELISA). Trimethylamine N-oxide (TMAO) content was determined by liquid chromatography with triple-quadrupole mass spectrometry. AMPK/sirtuin 1 (SIRT1) pathway-related proteins were detected using Western blotting.Results: AZI dose-dependently alleviated CS-induced lung dysfunction and colon tissue injury (p < 0.05), reduced the ratio of Th17/Treg, facilitated the expression levels of interleukin (IL)-10 and sIgA, and inhibited the levels of pro-inflammatory factors and TMAO (p < 0.05). In addition, AZI activated the AMPK pathway and upregulated SIRT1 expression in colon tissues (p < 0.05). Critically, the protective effects of AZI were partially reversed by co-treatment with the AMPK inhibitor Compound C (p < 0.05).Conclusion: AZI may regulate the Th17/Treg cell balance in association with TMAO reduction and AMPK/SIRT1 activation, thereby improving intestinal mucosal immunity in COPD rats. These findings highlight the potential of targeting the gut microbiota and mucosal immunity as a therapeutic strategy for COPD management.
Background: Recurrent hypoglycemia is a common complication of intensive glycemic control in patients with type 2 diabetes mellitus and may contribute to cardiovascular injury. However, the molecular mechanisms underlying hypoglycemia-induced myocardial inflammation remain incompletely understood. This study aimed to elucidate the function and mechanistic pathways by which the RNA-binding protein ELAV-like RNA binding protein 1 (ELAVL1, also termed Hu antigen R / HuR) modulates NLRP3 pathway-associated inflammatory responses in the context of myocardial injury exacerbated by recurrent hypoglycemia in type 2 diabetes mellitus.Methods: A total of eighteen male C57BL/6J mice were allocated at random into three experimental groups (n = 6 per group): a control group, a type 2 diabetes mellitus group generated by high-fat feeding followed by streptozotocin administration, and a diabetic hypoglycemia group, in which diabetic mice received daily intraperitoneal insulin treatment for five consecutive days to establish recurrent hypoglycaemic episodes. Hypoglycemia was defined as a blood glucose concentration of ≤3.5 mmol/L maintained for 1 h. Cardiac function was assessed by echocardiography. Myocardial histopathology was examined by hematoxylin–eosin staining. Serum concentrations of cardiac troponin I (cTnI), cTnT, interleukin-1β (IL-1β), IL-18, and insulin were quantified using enzyme-linked immunosorbent assay (ELISA). Myocardial ELAVL1 mRNA expression was quantified by quantitative real-time PCR (qPCR). An intermittent low-glucose/recovery model was established in HL-1 atrial cardiomyocytes. ELAVL1 expression was silenced (sh-ELAVL1) or overexpressed (oe-ELAVL1) using lentiviral vectors. Cell injury was evaluated by CCK-8 assay and lactate dehydrogenase (LDH) release. The transcript and protein abundance of ELAVL1, NLRP3, and cleaved Caspase-1 (p20) were determined using qPCR and immunoblot analysis, whereas GSDMD-N (p30) expression was visualized by immunofluorescence.Results: HDM mice demonstrated greater reductions in left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS) than diabetes mellitus (DM) controls (p < 0.05), with more severe myocardial fiber disorganization, interstitial edema, and inflammatory cell infiltration. The HDM group displayed markedly higher circulating concentrations of cTnI, cTnT, IL-1β, and IL-18, together with increased myocardial ELAVL1 transcript abundance (p < 0.05). In vitro, recurrent hypoglycemic stimulation reduced cell viability and increased LDH release (p < 0.05), accompanied by upregulation of ELAVL1 and NLRP3 expression. ELAVL1 silencing restored cell viability, reduced LDH release, and decreased the expression of NLRP3, cleaved Caspase-1 (p20), and GSDMD-N (p30) (p < 0.05), whereas ELAVL1 overexpression exerted the opposite effects (p < 0.05).Conclusion: Recurrent hypoglycemia exacerbates myocardial injury in diabetic mice and is accompanied by enhanced inflammatory responses and increased ELAVL1 levels. In vitro functional studies further demonstrated that ELAVL1 positively regulates NLRP3/Caspase-1/gasdermin D (GSDMD)-associated pyroptotic signaling and aggravates hypoglycemia-induced cardiomyocyte injury. These findings suggest that ELAVL1 may contribute to the pathogenesis of diabetes-associated hypoglycemic myocardial injury and represents a potential therapeutic target.
Background: Urolithin A (UA) is a safe and promising therapeutic strategy for aging-related disorders, including osteoporosis. Sirtuin 3 (SIRT3) has been reported as a potential modulator of bone-joint pathologies such as osteoclast activation. However, whether UA affects osteoclast activation and function through SIRT3 remains unclear. This study aims to elucidate the mechanisms by which UA alleviates osteonecrosis.Methods: Primary bone-marrow-derived macrophages (BMMs) were isolated from mice, treated with Macrophage Colony-Stimulating Factor (M-CSF) and Receptor Activator of Nuclear Factor-κB Ligand (RANKL). Mature osteoclasts were identified by Tartrate-Resistant Acid Phosphatase (TRAP) staining to assess differentiation. Cell viability was measured with Cell Counting Kit-8 (CCK-8) assays. The expression of osteoclast-related protein was detected by Western blot, and its mRNA level was detected by quantitative Real-Time polymerase chain reaction (qRT-PCR). The effects of UA on osteoclast mitophagy and mitochondrial respiration were assessed using Western blot and a Seahorse analyzer. Osteonecrosis of the femoral head was evaluated by hematoxylin and eosin staining (H&E), Safranin O/Fast Green, and TRAP staining. SIRT3 protein levels were assessed by Western blot.Results: In vitro, UA (3 μM) significantly decreased the number of TRAP-positive osteoclasts (p < 0.05) and downregulated the expression of osteoclast-related genes (Acid phosphatase 5 (Acp5), cathepsin K (Ctsk), Matrix metallopeptidase 9 (Mmp9), Integrin subunit beta 3 (Itgb3), ATPase H+ transporting V0 subunit d2 (Atp6v0d2); p < 0.05). UA treatment reduced SIRT3 protein levels (p < 0.05), elevated ATPase inhibitory factor 1 (ATPIF1) acetylation, and suppressed receptor-mediated mitophagy markers (BCL2/adenovirus E1B interacting protein 3-like (Nix) and BCL2/adenovirus E1B 19 kDa protein-interacting protein 3 (Bnip3); p < 0.05) in osteoclasts, while impairing mitochondrial respiration and increasing mitochondrial ROS production. SIRT3 overexpression partially reversed these effects. In vivo, UA administration attenuated cartilage thinning, trabecular collapse, and reduced TRAP-positive cells and SIRT3/Nuclear factor of activated T cells, cytoplasmic 1 (NFATc1) expression (p < 0.05) in a glucocorticoid-induced osteonecrosis mouse model.Conclusion: UA effectively inhibits osteoclast activation and alleviates glucocorticoid-induced osteonecrosis, and its protective effects are associated with downregulation of SIRT3 expression as well as the expression of mitophagy-related markers Nix and Bnip3. UA may represent a potential therapeutic candidate for osteoclast-mediated bone disorders.
Background: On most postoperative imaging examinations, atelectasis after lobectomy is a transient finding. We focused on the more clinically consequential scenario: persistent collapse of at least one segment of the lung beyond the first postoperative week. These cases may delay mobilisation, impair secretion clearance, and increase the need for additional airway care after video-assisted thoracoscopic (VATS) lobectomy for non-small cell lung cancer (NSCLC). This study investigated perioperative predictors of this specific endpoint and used the identified factors to develop a nomogram.Methods: We reviewed 1268 consecutive NSCLC patients treated with VATS lobectomy from January 2020 to December 2024. Persistent atelectasis meant radiographic collapse of at least one pulmonary segment for 7 or more postoperative days. Before modelling, 40 candidate predictors were coded. We then reviewed these predictors according to clinical domain, avoided retaining overlapping variables that represented the same construct, and performed 1000 bootstrap resamples for internal validation. Age, smoking exposure, forced expiratory volume in 1 second (FEV1), and operative time were additionally examined as continuous variables, and restricted cubic splines were used to assess possible nonlinear associations.Results: Persistent atelectasis was recorded in 368 patients (29.0%). Twenty non-redundant variables were included in the adjusted analysis, corresponding to 18.4 events per variable. The final score retained age ≥65 years, smoking history ≥20 pack-years, chronic obstructive pulmonary disease (COPD), FEV1 <60% predicted, upper lobe resection, and operative time ≥180 minutes (all p < 0.001). The apparent area under the receiver operating characteristic curve (AUC) was 0.842 (95% confidence interval (CI) 0.818–0.865); the bootstrap-corrected AUC was 0.837 (95% CI 0.813–0.861). Calibration was acceptable (Hosmer-Lemeshow chi-square = 5.41, p = 0.714; slope = 0.92). Observed rates were 10.2%, 38.1%, and 79.0% in the low-, intermediate-, and high-risk groups (p for trend <0.001). Sensitivity analyses using continuous predictors produced similar discrimination, and spline analyses showed significant overall associations for age, smoking exposure, FEV1, and operative time.Conclusion: In this cohort, persistent atelectasis was associated with pulmonary reserve, chronic airway disease, lobe removal, and the length of the operation. Because operative time is included in the score, the complete nomogram is best applied during perioperative or early postoperative care rather than preoperatively. Therefore, further multicentre validation remains necessary.
Background: Excisional procedures for high-risk human papillomavirus (HPV)-related lower genital tract lesions fail to eliminate the underlying HPV infection and carry fertility-related risks, leaving patients with persistent low- or high-grade lesions without an effective tissue-preserving option. Real-world data on 5-aminolevulinic acid photodynamic therapy (ALA-PDT) across the full lesion spectrum remain limited. This study evaluated composite and histological/cytological complete response rates of ALA-PDT and explored whether morphological and virological responses follow distinct temporal trajectories.Methods: In this single-center retrospective cohort study conducted between 2021 and 2024, 176 patients were included after excluding isolated vulvar intraepithelial neoplasia (VIN), isolated condylomata acuminata, and HPV infection <12 months without morphological lesions. A total of 177 treatment episodes were recorded, as one patient received two independent ALA-PDT courses separated by >6 months, and only the first episode was retained in the primary per-patient analysis. Of these, 174 patients had confirmed cervical or vaginal intraepithelial neoplasia (with or without concurrent vulvar lesions or condylomata acuminata), and 2 had persistent HPV infection without morphological lesions. All patients received standardized ALA-PDT (20% ALA; 635 nm; 120 J/cm2; 3–6 sessions). Of 174 patients with morphological disease, 147 had valid follow-up and constituted the primary efficacy population. The primary endpoint was composite complete response (CR), requiring concurrent morphological resolution, cytological normalization, and HPV clearance. A predefined secondary endpoint, histological/cytological CR (hCR), required morphological and cytological normalization irrespective of HPV status.Results: Among 147 evaluable patients with morphological disease, composite CR was 68.7% (101/147), with hCR of 83.0% (122/147) and overall response rate of 93.2% (137/147). These estimates were robust across follow-up strata (CR ≥6 months: 70.8%; ≥9 months: 73.3%) and after inverse probability weighting (68.1%). CIN 2/3 achieved the highest composite CR (85.7%). Cross-sectional observations suggested that cytological normalization may plateau earlier than HPV clearance, which continued to increase through 12 months.Conclusions: ALA-PDT demonstrated high efficacy across HPV-related lower genital tract lesions, with morphological response rates suggesting clinically meaningful disease control. These findings support ALA-PDT as an effective, tissue-preserving option for HPV-related lower genital tract lesions. Direct assessment of reproductive outcomes, sexual function, quality of life, and patient satisfaction was not feasible in this retrospective dataset and warrants future prospective studies.
Background: Alzheimer’s disease (AD) is frequently accompanied by type 2 diabetes mellitus (T2DM), and the two disorders share some common pathological mechanisms. However, the core genes responsible for this comorbidity and the therapeutic targets remain to be identified.Methods: Gene expression data were obtained from the Gene Expression Omnibus (GEO) database for AD (GSE5281) and T2DM (GSE76894), and differentially expressed genes (DEGs) were identified. Weighted gene co-expression network analysis (WGCNA) was performed to identify AD-associated modules. Four machine learning algorithms—Least Absolute Shrinkage and Selection Operator (LASSO) regression, Random Forest (RF), Support Vector Machine Recursive Feature Elimination (SVM-RFE), and Boruta—were used to screen core genes. Gene set enrichment analysis (GSEA), immune infiltration analysis using the CIBERSORT method, and drug prediction using the Comparative Toxicogenomics Database (CTD) were then conducted. In order to evaluate the binding mode and affinity of the predicted drugs with core targets, molecular docking was performed.Results: The number of DEGs identified in the AD dataset was 4970, and the number of DEGs identified in the T2DM dataset was 392. Cross-analysis of DEGs and WGCNA identified 15 overlapping disease-related genes. Two core genes, enolase 2 (ENO2) and membrane palmitoylated protein 1 (MPP1), were identified using four machine learning algorithms. Both genes were co-enriched in GSEA for leukocyte transendothelial migration, long-term potentiation/depression, citric acid cycle, oxidative phosphorylation, and glycolysis/gluconeogenesis. Immune infiltration analysis revealed that they participated in the disease process by regulating the immune microenvironment. Valproic acid (VPA) was selected as a candidate drug based on the results of the CTD drug prediction. The molecular docking results revealed that the binding free energies of VPA with ENO2 and MPP1 were –4.747 and –5.346, respectively, suggesting that VPA had a higher binding affinity for MPP1.Conclusion: ENO2 and MPP1 are shared molecular markers between AD and T2DM, and may be involved in the pathobiology of both diseases by regulating energy metabolism, synaptic function, and immune inflammation. VPA may exert potential therapeutic effects by targeting these two shared nodes, providing preliminary molecular evidence for the mechanisms underlying AD-T2DM comorbidity. Further validation in AD-T2DM comorbid cohorts is warranted.
