搜索结果：Tissue & cell

Endometrial cancer (EC) is the most common gynecologic malignancy, yet effective therapies for advanced and recurrent disease remain limited. Sodium butyrate (NaB), a gut microbiota-derived short-chain fatty acid (SCFA) with known anticancer activity, remains poorly understood in EC. In this study, we investigated the anticancer effects of NaB in two EC cell lines, HEC1A and AN3CA. NaB dose-dependently inhibited cell viability, colony formation, and migration in both cell lines, with HEC1A cells exhibiting greater sensitivity. NaB differentially modulated epithelial-mesenchymal transition (EMT)-related markers between the two cell lines. NaB markedly induced apoptosis in HEC1A cells, whereas AN3CA cells showed resistance to apoptotic cell death, despite mitochondrial membrane depolarization occurring in both cell lines. Cell cycle analysis revealed subG1-accumulation in HEC1A cells and G1 phase arrest in AN3CA cells. Notably, NaB potently suppressed thymidylate synthase (TS) at both mRNA and protein levels in both cell lines, representing the first demonstration of TS suppression by NaB in EC. NaB also broadly reprogrammed pyrimidine metabolism by downregulating de novo synthesis enzymes while upregulating salvage pathway components. Taken together, these findings suggest that TS suppression and pyrimidine metabolic reprogramming are important components of the in vitro response of EC cells to NaB. These results provide mechanistic insight into the metabolic effects of NaB in EC and support further investigation of this pathway in pharmacologically relevant and locally applicable therapeutic contexts.

Excitable cells are commonly studied via the extracellular potentials (EPs) they generate, which underlie signals in electroencephalography (EEG), electrocardiography (ECG), and multielectrode array (MEA) recordings. However, some excitable systems produce little or no detectable EPs, for reasons that remain poorly understood. Here we show mathematically that homogeneous excitable cells and tissues - with spatially uniform ion channel distributions and no external stimulation - are extracellularly silent during spatially uniform, non-propagating action potentials (i.e., in the absence of a traveling wavefront). Specifically, an isolated, autonomous cell with uniform membrane properties generates zero EP, independent of shape, kinetics, or model complexity. The result extends to coupled cells provided the tissue remains fully homogeneous. EPs emerge only from spatial inhomogeneities, propagating electrical waves, or applied currents. We demonstrate the physiological relevance of this principle in Purkinje neurons, where clustering of sodium channels enables ephaptic synchronization, while uniform cells remain asynchronous and undetectable extracellularly. We further show that connected human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) and pancreatic β-cells exhibit EPs in proportion to cellular or tissue-level heterogeneity.

Complex tissue architecture is achieved through multiple rounds of morphological transitions. Here, we analyzed cellular flows and tissue mechanics during avian skin development by employing chicken and transgenic quail skin explant models. We demonstrate how novel cellular flows initiate chemo-mechanical circuits that guide epithelial protrusion, folding, invagination, and spatial cell fate specification. During initial feather bud formation, stiff dermal condensates protrude vertically from the locally softened epithelial sheet. As the bud elongates, it stretches the epithelial cells at the base, thus mechanically activating YAP, which causes the epithelial sheet to fold downward and form a stiff cylindrical wall that invaginates into the skin. This stiff epithelial tongue is essential for the compaction and formation of the tightly packed dermal papillae. These topological transformational events are mechanically interconnected, and the completion of one circuit initiates the next. In contrast, during scale development, the rigid epithelial sheet restricts dermal cell flows, preventing further topological transformation. Based on these findings, we developed a topological transformation model describing how this process enabled the evolution of feather follicles from scales.

Despite being a standard treatment for locally advanced esophageal squamous cell carcinoma (ESCC), neoadjuvant chemoradiotherapy (nCRT) followed by surgery was frequently associated with toxicities. Body composition was a potential factor influencing both the toxicity and efficacy of nCRT. The aim of this study was therefore to assess, within a large retrospective cohort derived from prospective trials, the associations of body composition measures-including skeletal muscle index (SMI), skeletal muscle radiodensity (SMD), subcutaneous adipose tissue (SAT), and visceral adipose tissue (VAT) with nCRT-related toxicities and outcomes. Patients with locally advanced ESCC who underwent standard nCRT between January 2019 and December 2024 were included in this retrospective analysis. Body composition parameters were derived from pre-treatment computed tomography (CT) scans at the third lumbar vertebra (L3). SMD was calculated as the mean radiation attenuation (Hounsfield units) of the muscle area. Based on established criteria, patients were stratified as having low or high SMI according to Martin's BMI-specific thresholds, while SMD and VAT were categorized into tertiles (low, intermediate, high). Hematologic toxicities and dose-limiting toxicities (DLT) were graded using CTCAE v5.0. Associations between body composition and toxicities/efficacy were analyzed using logistic regression. The relationship between body composition and absolute lymphocyte count (ALC) nadir and baseline lymphocyte subsets (CD4+, CD8+, CD3+) was also evaluated. This study included 297 patients (median age 66 years; 85.5% male; 72.4% stage III). Following nCRT, 278 patients underwent surgery, achieving a pathological complete response (pCR) rate of 42.8% and tumor regression grade (TRG) 0-1 of 72.3%. Grade ≥2 hematologic toxicity occurred in 80.5% (n=239) of patients, and 35.4% (n=105) experienced DLT. While body composition showed no significant association with pCR or TRG, multivariate analysis revealed that both high SMI (adjusted odds ratio [OR] 0.48, 95% confidence interval [CI] 0.26-0.89, P=0.02) and high VAT (adjusted OR 0.46, 95% CI 0.22-0.94, P=0.034) were independent protective factors against grade ≥2 hematologic toxicity. Additionally, high SMI was significantly associated with reduced risk of DLT (adjusted OR 0.56, 95% CI 0.34-0.92, P=0.023). Notably, patients with low VAT had significantly lower median ALC nadir and baseline CD4+ T-cell counts. In this cohort, pretreatment high SMI and high VAT were associated with reduced nCRT-related hematologic toxicities (grade ≥2) and DLT in patients with locally advanced ESCC. Pathological response (pCR/TRG 0-1) was not associated with body composition. The protective role of high VAT might be partially mediated by its association with higher lymphocyte nadirs and baseline CD4+ T-cell counts. Future studies should investigate long-term outcomes and assess whether prehabilitation strategies targeting SMI and VAT can mitigate toxicity without compromising treatment efficacy.

Cold preservation is a critical logistical step in liver transplantation but induces ischemia-reperfusion injury (IRI), a key driver of early graft dysfunction. While bulk tissue assays capture global damage, they obscure the cell-type-specific transcriptional programs engaged during hypothermic storage. We utilized a multicellular human liver-on-chip model comprising Patient-Derived Organoids (PDOs), hepatic stellate cells (HSCs), liver sinusoidal endothelial cells (LSECs), and macrophages. Chips were exposed to 24-h static cold storage using either the clinical standard University of Wisconsin (UW) solution or a hyperbranched polyglycerol (HPG)-based formulation, followed by normothermic reperfusion. Single-cell RNA sequencing (scRNA-seq) was performed to map transcriptional trajectories across the preservation-reperfusion axis. We identified candidate solution-dependent transcriptional differences across cell types. PDOs from UW-preserved chips showed comparatively higher mean expression of inflammatory and oxidative stress-associated transcripts (IFI27, SAA1, HMOX1) and mitochondrially-encoded genes (MT-ND5) relative to HPG-preserved samples, which retained comparatively higher expression of homeostatic epithelial markers (EPCAM, KRT18). HSCs and LSECs in the UW group showed comparatively elevated expression of fibrosis-associated (COL1A1, TAGLN) and endothelial adhesion (ICAM1) transcripts. Ligand-receptor interaction modelling identified candidate inflammatory communication axes, including chemokine signaling interactions (CXCL1, CCL20) between macrophages and epithelial compartments, with higher predicted activity under UW preservation. This study provides an exploratory, high-resolution map of cell-type-specific transcriptional patterns associated with hypothermic preservation in a liver-on-chip model. Our findings suggest that preservation solution chemistry is associated with distinct transcriptional signatures spanning stress response, mitochondrial, and intercellular signaling pathways. Transcriptional patterns in HPG-preserved cells were consistent with comparatively attenuated injury responses; however, these observations are hypothesis-generating and require independent biological replication and functional validation, including metabolic flux assays and ROS production measurements before conclusions regarding mitochondrial protection or clinical preservation efficacy can be drawn.

Cytokine-mediated cross-talk between immune cells and fibroblasts is a driver of excessive ECM accumulation during fibrosis. In this study, we used a 3D in vitro model of a connective tissue to discern the roles of three pro-inflammatory cytokines; TNF-α, IL-18 and IL-1β, alone, and in combination with TGF-β1 to simulate the fibrotic environment. Ring-shaped tissues were formed by seeding human fibroblasts into circular molds of agarose, wherein the cells self-assembled, formed a 3D tissue and synthesized de novo a collagen-rich ECM. Cytokine treated tissues were analyzed at days 7 and 14 by histology and measured for thickness, collagen, DNA and strength and stiffness by tensile testing. Despite their pro-inflammatory nature, none of the cytokines increased collagen alone or in combination with TGF-β1. TNF-α and IL-1β reduced collagen, tissue strength and stiffness, and altered tissue morphology. When combined with TGF-β1, TNF-α and IL-1β counteracted TGF-β1-mediated increases in collagen, strength, and stiffness. In contrast, IL-18 had minimal effects alone or when combined with TGF-β1. These data suggest that IL-18 has no effect on fibroblast activation, whereas TNF-α and IL-1β may modulate TGF-β1's effects. This 3D model of a human collagen-rich tissue can help define cytokine-mediated cross-talk between immune cells and fibroblasts.

Micro- and nanoplastics have been recognized to pose potential threat for human environment and health. Plastic particles of different, sizes, forms and composition have already been detected in human tissue, but little information is available so far on how particle numbers are relating to cellular effects in dependence of size. We here set out to utilize a mast-cell/basophilic cell model to investigate the role of different polystyrene bead numbers and sizes on mast-cells under conditions that allow for local accumulation of auto- and paracrine released cytokines and mediators, resembling the conditions in inflamed tissue. Cultured RBL-2H3 cells responded to spherical polystyrene (PS) microplastic particles with degranulation and release of inflammatory mediators. Our results indicate a non-linear dose response curve for mast-cell mediator release, due to induction of apoptosis at high particle counts for all tested particle sizes. Cellular uptake of larger (1µm) particles was found to enhance necrotic cell death.

Pain is a hallmark of inflammation and tissue injury, particularly following major surgical procedures. Despite its prevalence and clinical impact, pathological pain, defined as persistent pain that impairs function, remains a major unmet medical need. Human birth tissue, including the amniotic membrane and umbilical cord, has long been recognized for its regenerative properties and its ability to support wound healing. Recent studies have identified heavy chain 1 (HC)-hyaluronic acid (HA)/pentraxin 3 (HC-HA/PTX3) as a key matrix component within these tissues. This complex exhibits anti-inflammatory and anti-scarring activities and helps maintain stem cell quiescence. Emerging evidence, including our own findings, indicates that human birth tissue-derived products may also modulate pain responses in certain settings, such as post-surgical pain, potentially through mechanisms involving neuronal inhibition and regenerative healing. We summarize the molecular and cellular mechanisms by which human birth tissue-derived products and HC-HA/PTX3 may exert anti-inflammatory and analgesic effects. We then highlight preclinical and clinical studies evaluating their potential roles in wound healing and pathological pain. Finally, we discuss translational opportunities, current challenges, and future directions for advancing these biologics within the emerging field of regenerative pain medicine. This review outlines a framework for potential regenerative pain management using birth tissue-derived products, which may serve as a foundation for developing new therapies for certain pathological pain conditions.

Benign breast disease (BBD) is common and confers heterogeneous increases in breast cancer risk; however, risk prediction relies mainly on histopathology and clinical factors. Sclerosing adenosis (SA) is a proliferative BBD lesion associated with an approximately two-fold increase in risk, yet most women with SA never develop breast cancer. We hypothesize that the immune-stromal microenvironment of SA and its surrounding lobular field relates to subsequent invasive breast cancer. In a nested case-control study within a BBD cohort, we profiled 24 sclerosing adenosis (SA) biopsies (9 developing invasive breast cancer within 15 years, cases; 15 cancer-free at ≥ 15 years, controls). We integrated whole-tissue NanoString gene-expression profiling with multiplex immunofluorescence (MxIF) imaging of SA lesions and surrounding morphologically normal lobules. We measured immune and stromal biomarkers in SA lesions and adjacent lobules, with image analysis masked to case-control status, integrated these data with whole-tissue gene expression, and summarized both microenvironment patterns and the proximity of immune cells to proliferating epithelium. SA biopsies from women who later developed cancer showed a low-immune, high-stromal gene-expression program, whereas controls were enriched for immune signatures. Stromal densities of CD8⁺, CD68⁺ and RUNX3⁺ cells in both lobular stroma and SA lesions mirrored this axis and were markedly lower in cases than controls. Unsupervised clustering identified immune-cold and immune-hot lobule types and four SA lesion field archetypes; immune-hot lobules and immune-hot/epithelium-proliferative lesion fields were enriched in controls. Spatial analyses further showed that immune-hot lobules have stromal immune cells positioned closer to proliferating epithelium and enriched CD27-CD8 microclusters, whereas SA lesions from cases exhibit greater immune-to-Ki67 distances, fewer boundary-proximal CD8⁺ sentinels, and depletion of CD27-RUNX3 and RUNX3-CD8 microclusters. These findings support an association of an immune-cold SA lesion embedded within an immune-cold lobular field phenotype with subsequent invasive breast cancer risk in women with SA, and suggest that spatially organized, RUNX3-rich immune microenvironments may contribute to epithelial surveillance. Validation in larger cohorts will be needed to confirm generalizability and clarify lesion-specific versus field-wide contributions.

Integrin-linked kinase (ILK) is a key oncogenic driver in oesophageal squamous cell carcinoma (ESCC). This study evaluated the antitumour effects of the novel ILK inhibitor Nilotinib and explored its downstream mechanisms. In vitro, TE-1 and KYSE150 cells were assessed using Cell Counting Kit-8, lactate dehydrogenase release, colony formation, 5-ehynyl-2 ' -deoxyuridine incorporation, flow cytometry, Transwell assays, and Western blotting to confirm ILK targeting and determine functional changes. Electron microscopy and fluorescent probes with flow cytometry were used to analyse mitochondrial alterations. In vivo, a nude mouse subcutaneous xenograft model was established to examine tumour growth after peritumoural Nilotinib administration; hematoxylin and eosin staining assessed tissue changes, and immunohistochemistry measured Ki67 and cleaved-caspase 3 expression. ILK overexpression alleviated Nilotinib-induced cytotoxicity, restored proliferation, increased proliferating cell nuclear antigen (PCNA) and Ki67, and reduced cleaved-caspase 3 and cleaved poly(ADP-ribose) polymerase (PARP), supporting ILK as a primary target. Nilotinib dose-dependently inhibited proliferation, invasion, and metastasis while promoting apoptosis, accompanied by downregulation of PCNA, Ki67, [matrix metalloproteinase 2 (MMP2), MMP9, and COX2] and upregulation of cleaved-caspase 3 and cleaved-PARP. In xenografts, Nilotinib significantly reduced tumour size and weight, decreased Ki67, and increased cleaved-caspase 3.RNA sequencing identified autoimmune regulator (AIRE) as a markedly downregulated molecule following Nilotinib treatment. Cycloheximide chase assays indicated accelerated AIRE protein degradation, while MG132 partially rescued AIRE levels, implicating proteasome-dependent degradation. Overall, Nilotinib suppresses ESCC progression by inhibiting ILK and destabilising AIRE, suggesting its potential as a targeted therapy for ILK-positive ESCC.

Osteoarthritis is a chronic disabling disease characterized by progressive degeneration of articular cartilage. The condition is characterized by an imbalance in the inflammatory response, the degradation of the extracellular matrix, the apoptosis of chondrocytes, and the disorder of the immune microenvironment. The present clinical efficacy of the treatment is unsatisfactory due to the limited self-healing ability of the articular cartilage. In recent years, there has been a growing body of research focusing on the therapeutic potential of exosomes (Exos) derived from mesenchymal stem cells (MSCs). The findings of these studies have demonstrated the significant potential of MSC-derived exosomes as a promising cell-free therapeutic strategy. This article provides a synopsis of the research progress and mechanisms of mesenchymal stem cell-derived exosomes (MSC-Exos) from diverse tissue sources in the treatment of osteoarthritis. Furthermore, the application of MSC-Exos in the treatment of osteoarthritis were also discussed. The review will provide a valuable reference for future research directions in this field.

Connexin 43 (Cx43) exhibits remarkable functional diversity that is precisely dictated by its dynamic subcellular localization. Beyond its canonical role at the plasma membrane, where it assembles into gap junctions (GJs) and hemichannels (HCs) to mediate intercellular communication, Cx43 translocates to the nucleus and mitochondria, where it exerts non-channel functions including transcriptional regulation and metabolic adaptation. At the plasma membrane, dysregulation of Cx43 trafficking, anchoring, or turnover leads to excessive HC opening and impaired GJ communication, contributing to cardiovascular arrhythmias, ischemia-reperfusion injury, neuroinflammation, osteoporosis, and retinopathy. In the nucleus, Cx43 or its C-terminal fragment enters through importin-dependent pathways, functioning as a non-canonical transcriptional regulator; its mislocalization is implicated in cancer (context-dependent suppression or promotion), hepatic gluconeogenesis in diabetes, and tissue fibrosis. Within mitochondria, Cx43 is imported via Hsp90/TOM complex- or GJA1-20 k-dependent pathways, where it regulates K+ transport, respiratory chain activity, and redox balance; this mitochondrial pool exerts cardioprotection under preconditioning but exacerbates diabetic cardiomyopathy and neurological injury under pathological stress. This review synthesizes current knowledge on the trafficking mechanisms, pathological outcomes, and therapeutic targeting of Cx43 in these three subcellular compartments. We further discuss peptide-based inhibitors (e.g., Gap19, αCT1), small molecules (e.g., tonabersat, danegaptide), and natural product-derived modulators, highlighting challenges in specificity, bioavailability, and clinical translation. By linking compartment-specific functions to distinct disease entities, this review establishes subcellular localization as a central determinant of Cx43 biology and a promising axis for precision medicine.

Danshensu (DSS) is one of the water-soluble components extractable from the traditional Chinese medicine Salvia miltiorrhiza Bge., exhibiting pharmacological effects such as promoting blood circulation, dilating coronary arteries, and improving cerebral blood flow. The Danshensu derivative (OZD-1) obtained through the derivatization of DSS is a potential multi-target drug for the central nervous system, however, its mechanism of action against cerebral ischemia-reperfusion injury (CIRI) remains unclear. Systematically investigating the therapeutic potential and mechanisms of action of Danshensu derivative against cerebral ischemia-reperfusion injury. Rat brain microvascular endothelial cells (RBMVECs) were cultured in vitro to establish an oxygen-glucose deprivation/reoxygenation (OGD/R) injury model. Groups included a control group, an OGD/R model group, and OZD-1 low-dose (12.5 μmol/L), medium-dose (25 μmol/L), and high-dose (50 μmol/L) groups. Cell viability, migration capacity, and vascular lumen formation were assessed using the CCK-8 assay, cell scratch assay, and matrigel matrix gel assay, respectively. In vivo, a transient middle cerebral artery occlusion (tMCAO) model was established in rats. Animals were randomly divided into the sham, model, OZD-1 (35, 70, 140 mg/kg), Edaravone (Eda), and DSS groups. Daily oral administration was performed post-surgery for 14 consecutive days. Tissue pathology staining, behavioral tests, and regional cerebral blood flow imaging assessed brain tissue damage, cognitive function, and ischemic side cerebral blood flow recovery, respectively. Transcriptome sequencing analyzed differential gene expression and pathway enrichment patterns. Western blot detection measured expression levels of proteins related to the PI3K-AKT-CREB signaling pathway, phosphoproteins, downstream apoptosis-related proteins, and CD31, CD34, and VEGFA proteins. In vitro experiments demonstrated that OZD-1 dose-dependently enhanced the viability of RBMVECs following OGD/R injury, significantly improving cell migration and luminal formation capabilities. In vivo studies revealed that compared to the model group, rats in all OZD-1 dosage groups exhibited markedly improved cognitive function, significantly restored cerebral blood flow in the ischemic hemisphere, and substantially reduced pathological brain tissue damage. Transcriptome sequencing results indicated significant enrichment of genes associated with the PI3K-AKT signaling pathway following OZD-1 intervention. Western blot experiments confirmed that OZD-1 significantly upregulates the phosphorylation levels of proteins related to the PI3K-AKT-CREB signaling pathway in OGD/R-injured cells and brain tissue from tMCAO rats, thereby promoting VEGFA-mediated angiogenesis and inhibiting apoptosis. To further verify pathway involvement, in vitro inhibition experiments were performed in RBMVECs using the PI3K inhibitor LY294002 and CREB inhibitor 666-15. These inhibitors abolished the OZD-1-induced upregulation of p-PI3K, p-AKT, and p-CREB, and reversed its protective effects on cell viability, migration, and tube formation. These results confirm that OZD-1 protects vascular endothelial cells directly via activating the PI3K-AKT-CREB pathway. OZD-1 exhibits significant neuroprotective effects against CIRI in rats, improving cognitive function, promoting vascular regeneration in ischemic areas, repairing damaged RBMVECs, and reducing apoptosis. Its mechanism of action is associated with the activation of the PI3K-AKT-CREB-VEGFA signaling pathway.

The umbilical cord and the amniotic membrane are a precious source of human mesenchymal stem cells (hMSCs), even though they are often discarded after the delivery. Due to their immunomodulatory and anti-inflammatory properties, hMSCs could be part of relevant strategies in the field of regenerative medicine. Additionally, they can be obtained from these tissues via a non-invasive and cost-effective process, overcoming ethical controversies. This study aims to propose protocol refinement to obtain hMSCs from the amnion and umbilical cord of healthy donors (utilizing limited and defined laboratory resources), and to compare and characterize these cells, thereby enabling future research on their properties. 30 women from "Paolo Giaccone" University Hospital of Palermo (Italy) were enrolled, according to the inclusion and exclusion criteria approved by the local Ethics Committee. A sample of umbilical cord and amnion was obtained from every patient and processed via an enzymatic or mechanical method. After refining the isolation protocol, hMSCs were characterized using flow cytometry, RT-qPCR, inducing a trilinear differentiation, and testing the formation of spheroids. This research shows reliable and practical methods to isolate hMSCs from birth tissues, validating them with extensive cell characterization. No direct association was observed between mothers' age and newborns' sex and the success rate in establishing hMSCs primary cultures, while a possible association between neonatal weight and the successful establishment of umbilical cord-derived cultures was found. Moreover, a difference in the adipogenic potential of the two hMSCs sources was highlighted. hMSCs have a relevant role in biomedicine, along with their derivatives, for their promising regenerative properties: this study aims to explore new insights to promote further research in this field.

Spatial transcriptomics extends traditional transcriptomic methods by quantifying gene expression within intact tissues while preserving each cell's precise spatial context. This technology also captures gene expression under physiological conditions, including interactions with the surrounding microenvironment, thereby enhancing our understanding of cellular states in both health and disease. Rapid recent advances have improved throughput, transcript capture, accuracy, and overall data quality. In this review, we summarize the major spatial transcriptomics platforms and outline their strengths and limitations. We also highlight key applications in neuroscience, including brain cell-type identification, structure-function relationships, and developmental processes. Additionally, we examine spatial gene-expression patterns in psychiatric and neurodegenerative disorders such as Alzheimer's disease and depression. Finally, we discuss emerging directions, including spatial multi-omics integration and the potential for artificial intelligence to advance brain research. Collectively, this work provides a foundation for future studies in neuroscience and brain disorders.

Bone remodelling is essential for maintaining skeletal integrity by preserving the balance between bone formation and resorption, with excessive osteoclast activity contributing to osteoporosis. Osteocytes act as central regulators of osteoclastogenesis through mechanically sensitive paracrine signals, yet the influence of osteoblasts and their mesenchymal precursors remains less defined. Extracellular vesicles (EVs) have recently emerged as mediators of bone cell communication, although their role in osteoclast regulation are still underexplored. This study demonstrates that mesenchymal-derived bone cells inhibit osteoclastogenesis through an EV-dependent mechanism shaped by their differentiation stage and mechanical environment. Mechanically stimulated osteocyte-derived EVs showed the strongest anti-catabolic response. Notably, we identify miR-150-5p as a mechano-responsive miRNA enriched within osteocyte EVs, capable of inducing a dose-dependent reduction in osteoclastogenesis. Transcriptomic analyses reveal that EV treatment and miR-150-5p delivery induce substantial transcriptional changes in osteoclast precursors, including downregulation of shared target genes linked to bone remodelling. Overall, we highlight mechanically activated osteocytes as key regulators of osteoclastogenesis through an EV-mediated mechanism, in which miR-150-5p represents a promising candidate contributor within the broader EV cargo landscape, highlighting their potential for future cell-free therapeutic strategies.

Accurate detection of KRAS codon mutations is essential for precision oncology in colorectal cancer (CRC), yet conventional liquid biopsy methods often lack sufficient sensitivity for rare ctDNA variants, particularly in early diseases. We developed a three-dimensional (3D) plasmonic KRAS microarray integrating blocked recombinase polymerase amplification with plasmon-enhanced fluorescence. Quencher-modified blocking probes suppress wild-type DNA while selectively enabling mutant signal amplification. A single primer-probe set per codon allows comprehensive detection of all substitutions within KRAS codons 12/13, 61, and 146. The platform achieved detection down to 1 fM by direct hybridization and 100 zM after blocked amplification, exceeding conventional PCR and next-generation sequencing sensitivity. Codon-level specificity was validated in CRC cell lines, with distinct signals for each mutation. Clinical analysis of 58 patients showed 100% concordance between tissue, plasma, and urine in mutation-positive malignant cases when sufficient input was available, indicating accurate reflection of tumor profiles. In benign tumors, detection was rare despite tissue mutations, likely due to limited ctDNA release.This plasmonic microarray enables ultra-sensitive, specific, and non-invasive detection, supporting early diagnosis, minimal residual disease monitoring, and longitudinal CRC management.

The metabolic enzyme lactate dehydrogenase C4 (LDHC4) is aberrantly expressed in cancers and linked to poor prognosis. However, its role in lung adenocarcinoma (LUAD) and the molecular mechanisms beyond glycolysis remain unclear. This study investigates whether LDHC4 promotes LUAD by modulating protein lactylation, a lactate-derived post-translational modification, focusing on the tumor suppressor retinoblastoma protein (RB1). LDHC4 expression and its correlation with clinicopathological features and survival were analyzed using public databases (UALCAN, Kaplan-Meier Plotter, LOGpc) and validated in a cohort of 90 paired LUAD tissues via immunohistochemistry. The functional impact of LDHC4 on proliferation, migration, and invasion was assessed in A549 and PC-9 cells using gain- and loss-of-function models. The global lactylation profile was analyzed using DIA-based lactylation proteomics on the Astral platform. The interaction between RB1 and E2F1 (E2F transcription factor 1) was examined through molecular dynamics simulations, co-immunoprecipitation (Co-IP), and immunofluorescence. The functional consequences of site-specific RB1 lactylation at lysine 900 (RB1-K900lac) were determined using RB1-K900R mutant constructs and cell cycle analysis. LDHC4 was significantly overexpressed in LUAD tissues, correlating with poor patient survival, and was an independent prognostic risk factor. In vitro, LDHC4 promoted LUAD cell proliferation, migration, and invasion, and its tumor-promoting role was corroborated in an LUAD xenograft model, in which derived tumors exhibited increased volume and weight compared with mock-transfected controls. Mechanistically, LDHC4 overexpression elevated global protein lactylation levels and specifically increased lactylation of RB1. Bioinformatics and molecular dynamics simulations identified K900 as a key conserved residue for RB1-E2F1 binding; its lactylation destabilized the complex by increasing structural fluctuation and weakening intermolecular interactions. Cellular experiments confirmed that the lactylation-resistant RB1-K900R mutant bound E2F1 more strongly than wild-type RB1. Functionally, cells expressing RB1-K900R exhibited suppressed malignant phenotypes and G1/S cell cycle arrest, accompanied by downregulation of CDKs/cyclins and upregulation of P21. This study uncovers a novel LDHC4-driven oncogenic axis in LUAD. LDHC4 facilitates RB1 lactylation at the K900 residue, which disrupts the RB1-E2F1 tumor-suppressive complex, leading to cell cycle dysregulation and tumor progression. These findings may position the "LDHC4-RB1 lactylation" axis as a promising therapeutic target for LUAD.

The kappa class of glutathione S-transferases 1 (GSTK1) is a vital regulatory factor in metabolic diseases. This study was conducted to investigate the regulatory effects of GSTK1 on renal ectopic fat deposition (EFD) and lipotoxic injury in diabetic nephropathy (DN) . HK-2 cells under high glucose(HG) / high fatty acid (HFA) stimulation, diabetic mice and human renal biopsy tissues were used. GSTK1 plasmid, GSTK1 siRNA and OSBPL8 siRNA were applied in vitro. Lipid accumulation was analyzed in the renal tissue of type 2 DN patients, diabetic mice and HK-2 cells under HG/HFA stimulation. The expression of GSTK1, DGAT1, ACAT1, CPT-1, BECLIN1, LC3II, ATG5 and RAB7 in renal tubular cells of diabetic mice and HK-2 cells under HG/HFA condition decreased significantly. Metformin treatment restored the expression of GSTK1 in diabetic mice. Additionally, the GSTK1 pharmacological modulator metformin relieved lipophagy dysfunction and promoted fatty acid (FA) β-oxidation enzyme CPT-1. In vitro, GSTK1 plasmid reduced lipid accumulation, fibrosis and inflammation and up-regulated the expression of CPT1 in HK-2 cells, but GSTK1 plasmid had no effect on lipid metabolizing enzymes (ACAT1, DGAT1) . In addition, GSTK1 plasmid could obviously restore lipophagy. However, pretreatment of HK-2 cells with the AMPK inhibitor Compound C, GSTK1 siRNA or OSBPL8 siRNA negated the activating effects of GSTK1 on lipophagy. This study indicated that GSTK1 could contribute to alleviate EFD in DN tubular cell through increasing the expression of FA β-oxidation enzyme CPT-1 and restoring lipophagy via AMPK-OSBPL8 pathway.