BACKGROUND:Hepatocellular carcinoma (HCC) is a leading cause of cancer death worldwide. Gemcitabine (Gem) is a commonly used drug against HCC, but its efficacy is limited by the development of resistance. Resveratrol (Res), a natural polyphenol with antitumor activity, may reverse Gem resistance in HCC, although the mechanism remains unclear. METHODS:The effects of Res on the proliferation, apoptosis, cell cycle, and invasion of Hep3B and HuH-7 cells were assessed via cell counting kit-8 (CCK-8), clonogenic, flow cytometry, and Transwell assays, respectively. Potential Res targets were predicted by network pharmacology, and markers of HCC prognosis were identified from the cancer genome atlas (TCGA) data. The interaction between Res and thymidylate synthase (TYMS) was validated by molecular docking and dynamics simulation. A Gem-resistant HuH-7 cell line (HuH-7/GR) was established, and when these cells were treated with Res combined with Gem, the effect on Gem sensitivity was detected by CCK-8 assay, clonogenic assay, and flow cytometry. Finally, a subcutaneous nude mouse model of HCC was used to evaluate the in vivo effects of Res combined with Gem. RESULTS:Res inhibited HCC cell proliferation, induced apoptosis and G2/M arrest, and suppressed invasion in a concentration-dependent manner. Network pharmacology and TCGA analysis identified TYMS as an important target gene for Res. TYMS was highly expressed in HCC tissues and correlated with poor prognosis. Res treatment reduced TYMS expression, while molecular docking and simulation showed stable binding of Res to TYMS. TYMS levels were elevated in HuH-7/GR resistant cells. Res combined with Gem was found to reverse drug resistance, inhibit proliferation and colony formation, and induce apoptosis. The Res + Gem combination group showed the smallest tumor volume in the in vivo model. CONCLUSION:By attenuating Gem resistance through TYMS inhibition, Res holds promise as a clinically viable adjunct to Gem-based chemotherapy, offering a potential strategy to improve outcomes in HCC patients.
BACKGROUND:Multipotent mesenchymal stromal cells (MSCs) are widely used in regenerative medicine. Their decellularized extracellular matrix (dECM) has emerged as a key bioactive substrate for tissue engineering. The properties of MSC-derived dECM depend on cell culture conditions. Physiological hypoxia mimics the native MSC milieu and represents a promising preconditioning strategy. This study aimed to evaluate the structural and mechanical properties of dECM derived from MSCs cultured under different oxygen levels and to assess the phenotype of MSCs recellularized on these matrices. METHODS:Human adipose-derived MSCs were permanently expanded under 20% or 5% O2 to obtain 20-dECM and 5-dECM. Matrix architecture was visualized using scanning electron microscopy and quantitative phase contrast. Stiffness of dECM was measured by atomic force microscopy. MSCs were recellularized on dECM and cultured under 20% O2 for 72 hours. The assessment of cell morphology, cytoskeletal organization, nuclear translocation of Yes-associated protein (YAP) and transcriptional coactivator with PDZ-binding motif (TAZ), integrin expression, levels of reactive oxygen species (ROS), and cell cycle analysis were assessed using confocal microscopy and flow cytometry. RESULTS:In comparison with 20-dECM, 5-dECM demonstrated significant alignment of fibrillar bundles and a 1.6-fold increase in stiffness (p < 0.05). Reseeded MSCs on 5-dECM have acquired a spindle-shaped, aligned morphology, whereas cells on 20-dECM displayed a rounded morphology with multiple processes. The YAP and TAZ nuclear-to-cytoplasmic ratio was significantly lower on both dECM compared with uncoated surfaces (p < 0.0001). Total fluorescence intensity of YAP and TAZ per cell revealed divergent patterns on 5-dECM vs 20-dECM: YAP levels were increased, whereas TAZ levels were decreased. The expression of integrin α2 and α6 was higher (p < 0.01), while the expression of integrin α1 and intercellular adhesion molecule 1 (ICAM-1) was lower on both dECM vs control (p < 0.05). Intracellular ROS levels were reduced twofold on dECM (p < 0.01). The proportion of MSCs in the G2/M phase and the increase in cell number were more significant on dECM compared with uncoated plastic. CONCLUSIONS:Hypoxia-derived dECM exhibits increased anisotropy and stiffness, promoting MSC alignment and cytoskeleton organization, integrin expression, altering the subcellular localization of YAP and TAZ and reducing ROS levels. Thus, it may represent a promising, effective bioactive scaffold for MSC expansion.
BACKGROUND:Glioblastoma is the most aggressive primary malignant brain tumor of the central nervous system and remains difficult to treat because of rapid progression, diffuse invasion, and frequent recurrence. This study aimed to identify a potential natural compound against glioblastoma using a reverse network pharmacology strategy and to investigate its anti-glioma effects and underlying mechanism. METHODS:Glioblastoma-related targets were collected from public disease databases and integrated with differentially expressed genes from a public transcriptomic dataset to identify intersecting targets. Protein-protein interaction analysis, hub target screening, functional enrichment analysis, and reverse network pharmacology were performed to identify candidate compounds. Molecular docking and 100-ns molecular dynamics simulations were used to assess interactions between 20-hydroxyecdysone and pathway-related targets. Cell viability, colony formation, apoptosis, migration, and protein expression were examined in U251 and U87 glioma cells. Statistical differences were analyzed using a paired Student's t-test for two-group comparisons or one-way analysis of variance (ANOVA) followed by Dunnett's multiple-comparisons test for multi-group comparisons, as appropriate. RESULTS:A total of 136 intersecting targets and 18 hub targets were identified. Functional enrichment analysis indicated that the phosphatidylinositol 3-kinase/protein kinase B signaling pathway was one of the key enriched pathways. Reverse network pharmacology identified 20-hydroxyecdysone as a candidate compound. Molecular docking showed relatively strong binding of 20-hydroxyecdysone to epidermal growth factor receptor, fms-related receptor tyrosine kinase 1, and integrin subunit alpha 5, and molecular dynamics simulations supported the stability of these complexes. In vitro experiments showed that 20-hydroxyecdysone inhibited cell viability, clonogenicity, and migration, while promoting apoptosis. It also reduced the phosphorylation levels of phosphatidylinositol 3-kinase, protein kinase B, and glycogen synthase kinase 3 beta at Ser9 without markedly altering total protein expression. CONCLUSION:Treatment with 20-Hydroxyecdysone showed anti-glioma activity in vitro and may exert its effects, at least in part, through suppression of the phosphatidylinositol 3-kinase/protein kinase B signaling pathway. These findings support its further evaluation as a potential therapeutic candidate for glioblastoma.
BACKGROUND:PDE4DIP encodes a scaffold protein that has been implicated in compartmentalized signaling and cytoskeletal organization, but the role of its myomegalin variant 8 (MMG8) isoform in hepatocellular carcinoma (HCC) remains unclear. To address this gap, we examined PDE4DIP expression in public HCC datasets and investigated the functional role of MMG8 in HCC models. METHODS:PDE4DIP expression was analyzed in The Cancer Genome Atlas Liver Hepatocellular Carcinoma (TCGA-LIHC) cohort and two Gene Expression Omnibus (GEO) cohorts (GSE14520, GSE36376). MMG8 function was assessed in Huh7 cells using siRNA-mediated knockdown and in Hepa1-6 cells using lentiviral Clustered Regularly Interspaced Short Palindromic Repeats - CRISPR-associated protein 9 (CRISPR-Cas9)-mediated knockout. Cell proliferation in MMG8-KD Huh7 cells and MMG8-KO Hepa1-6 cells was assessed using Cell Counting Kit-8 (CCK-8) assays, while Huh7 cell migration was evaluated using Transwell assays. Tumor growth was assessed using a murine subcutaneous tumor model. Immunohistochemical staining for Ki67 and cleaved caspase-3 was employed to assess tumor cell proliferation and apoptosis-associated changes, respectively. Gene set enrichment analysis was performed in TCGA-LIHC tumors stratified based on PDE4DIP expression. RESULTS:PDE4DIP expression differed between tumor and non-tumor tissues across HCC cohorts, although the directionality of this difference was not uniform. MMG8 knockdown in Huh7 cells reduced proliferation and migratory activity. A single-cell-derived MMG8-KO Hepa1-6 clone exhibited reduced proliferation in vitro and formed smaller tumors in vivo, with lower Ki67 positivity but no significant difference in cleaved caspase-3 positivity between groups. In tumors from the TCGA-LIHC cohort, PDE4DIP expression was associated with distinct transcriptional programs. Specifically, PDE4DIP-high tumors presented with positive normalized enrichment score (NES) values for several metabolic pathways, whereas adhesion/extracellular matrix (ECM), cell cycle/proliferation, and translation/ribosome-related pathways exhibited negative NES values. CONCLUSIONS:These findings support a functional contribution of MMG8 to proliferative, migratory, and tumor-growth phenotypes in the tested HCC models. Bulk gene-level PDE4DIP expression in human tumors was associated with context-dependent transcriptional states and should not be interpreted as a direct surrogate for MMG8 function.
BACKGROUND:Small cell lung carcinoma (SCLC) is a highly aggressive, rapidly proliferating malignancy largely corresponding to immune-cold tumor type. The role of the key oncogene c-Myc in regulating the complex immune microenvironment of SCLC remains unknown. METHODS:Immunohistochemistry and multiplex immunofluorescence techniques were used to analyze c-Myc expression and immune cell infiltration. c-Myc and β-catenin expression were analyzed using western blotting. Flow cytometry was used to quantify the expression of immunosuppressive ligands-including programmed death ligand 1 (PD-L1), CD47, CD155, and human leukocyte antigen-E (HLA-E)-and characterize the function of T cells and macrophages. Cytokine levels were evaluated using enzyme-linked immunosorbent assays. RESULTS:c-Myc expression in high tumor-infiltrating lymphocyte infiltration (TILhigh) tumor exceeded that in TILlow tumors (n = 24) and was positively correlated with CD163+ macrophage infiltration (p = 0.0180). Immunosuppressive ligands were universally expressed on SCLC cells. 10058-F4-a c-Myc inhibitor-substantially downregulated the expression of PD-L1, CD47, and CD155 and upregulated HLA-E expression. In the coculture system, the inhibition of c-Myc in SCLC cells significantly upregulated interferon-gamma production in Jurkat cells, inhibited macrophage phagocytosis, and decreased cytokine concentrations in the supernatant. Further mechanistic studies revealed that c-Myc inhibition modulated the Notch signaling pathway and reduced the expression of E-cadherin, vascular endothelial (VE)-cadherin, and β-catenin. A β-catenin agonist improved the regulatory effect of 10058-F4 on the immunosuppressive ligands in SCLC cells. CONCLUSIONS:c-Myc expression in SCLC is associated with immune infiltration and may regulate the expression of immune-related ligands on SCLC cells via WNT/β-catenin signaling.
BACKGROUND:To investigate the protective role of dexmedetomidine (DEX) in sepsis-associated acute lung injury through the regulation of macrophage ferroptosis. METHODS:In silico screening of GeneCards and FerrDb Vv3 databases was performed to identify ferroptosis-related candidate genes modulated by dexmedetomidineDEX, lung injury, and macrophage activity, prioritizing nuclear factor erythroid 2-related factor 2 (Nrf2)/Nfe2l2 for subsequent experimental validation. A murine cecal ligation and puncture model of acute lung injury (ALI) and lipopolysaccharide-stimulated macrophages were established and subjected to treatment with DEX and/or the Nrf2 inhibitor ML385. Glutathione peroxidase 4 (GPX4), Nrf2, and heme oxygenase-1 (HO-1) protein levels were analyzed using western blotting. Ferroptosis and oxidative stress markers (superoxide dismutase [SOD], glutathione [GSH], iron content, reactive oxygen species [ROS], malondialdehyde [MDA]) were measured. Samples of fresh lung tissue were observed by electron microscopy. Macrophage phenotypes and tumor necrosis factor alpha (TNF-α) expression were assessed. Macrophage-conditioned medium was used to culture non-small cell adenocarcinoma epithelial cells (A549), and cell migration was evaluated. Proliferation-related proteins (matrix metalloprotease 9 [MMP-9] and Snail-1), key tight junction proteins (zonula occludens-1 [ZO-1] and occludin) and the apoptotic status were also analyzed. RESULTS:DEX significantly reduced ROS, iron content, and MDA levels while concomitantly increasing SOD activity and GSH content in both ALI model mice and LPS-treated macrophages. DEX inhibited pro-inflammatory M1 macrophage polarization with a concomitant decrease in TNF-α release through Nrf2-dependent mechanisms. DEX was found to mitigate inflammatory microenvironment-induced macrophage ferroptosis through activation of Nrf2 signaling, thereby reducing damage in A549 cells. This effect may be linked to the regulation of macrophage inflammatory phenotypes. CONCLUSIONS:DEX improved sepsis-associated ALI by inhibiting macrophage ferroptosis and polarization toward the M1 type by activation of the Nrf2/HO-1 pathway.
BACKGROUND:Menopause induces estrogen deficiency and promotes a pro-inflammatory and pro-fibrotic cardiovascular environment that increases susceptibility to adverse remodeling after myocardial infarction (MI). This study evaluated resveratrol as an inflammatory modulator and assessed its effects on cardiac fibrosis and hypertrophy in aged ovariectomized Wistar rats with chronic MI. METHODS:Thirty-six, one-year-old female Wistar rats (450-500 g) were allocated to six groups (n = 6 per group), with or without resveratrol treatment: (1) Sham, (2) Sham-R, (3) Bilateral ovariectomy (OVX), (4) OVX-R, (5) OVX + left anterior descending coronary artery ligation (LADL); OVX-LADL, and (6) OVX-LADL-R. Structural alterations, collagen deposition, and inflammatory profiles were assessed using histological analysis, morphometric measurements, and correlation-based heatmap. LADL in ovariectomized rats resulted in marked structural disorganization, increased cardiac fibrosis, and a predominance of pro-inflammatory mediators. RESULTS:Resveratrol treatment attenuated collagen deposition, partially preserved tissue architecture, and shifted the inflammatory profile towards a less pro-fibrotic pattern. Heatmap analysis showed that resveratrol reduced the strength of correlations between inflammatory markers and fibrotic remodeling, particularly in estrogen-deficient animals. CONCLUSION:These findings suggest that resveratrol attenuates adverse cardiac remodeling after MI in ovariectomized rats by modulating inflammation-driven fibrosis under estrogen-deficient conditions a response associated with increased IFN-γ.
BACKGROUND:Calcium signaling plays a critical role in plant growth, development, and stress adaptation. However, the tissue-specific mechanisms governing calcium homeostasis in medicinal plants adapted to high-calcium karst environments remain poorly understood. This study investigated the functional divergence and potential molecular basis of calcium homeostasis in leaves and root tips of Sophora tonkinensis under pharmacological perturbation. METHODS:Treatments included lanthanum chloride (LaCl3) (a plasma membrane Ca2+ channel blocker), sodium orthovanadate (Na3VO4) (a Ca2+-ATPase inhibitor), and ethylene glycol-bis(β-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA) (an extracellular Ca2+ chelator). Subcellular calcium distribution and expression of 14 calcium signaling-related genes were analyzed at 20, 40, and 60 days post-treatment using potassium pyroantimonate precipitation in conjunction with transmission electron microscopy and quantitative real-time PCR. RESULTS:LaCl3 treatment initially induced intracellular calcium accumulation, followed by the engagement of alternative efflux pathways in leaves and increased extracellular deposition in root tips. Na3VO4 treatment exacerbated intracellular calcium overload, resulting in sustained calcium toxicity in leaves, whereas root tips exhibited coordinated calcium redistribution. EGTA treatment dampened overall calcium signaling. However, root tips maintained functionality by mobilizing internal calcium stores. Gene expression analysis revealed a significant upregulation of Sophora tonkinensis calmodulin-like protein 9 (StCML9) in roots (up to 11.7-fold relative to the control), with expression closely synchronized with Ca2+ fluctuations, suggesting a potential role for StCML9 in calcium sensing and adaptive responses. In contrast, Sophora tonkinensis calcium-dependent protein kinases (StCDPKs) were generally suppressed in leaves. CONCLUSIONS:These findings suggest that S. tonkinensis may employ organ-specific calcium distribution and transcriptional reprogramming to maintain calcium homeostasis under signaling perturbation. This study provides correlative evidence that contributes to mechanistic insights into the species' adaptation to high-calcium karst environments and offers candidate genes for future functional validation.
Septic cardiomyopathy (SCM) is a prevalent and serious cardiac complication arising from sepsis-induced multiple organ dysfunction syndrome (MODS). The pathogenesis of SCM is complex and primarily involves immune-inflammatory responses, oxidative stress, programmed cell death, and mitochondrial dysfunction. In recent years, mitochondrial quality control (MQC) has attracted growing interest as a central mechanism for maintaining cellular homeostasis and myocardial energy metabolism in SCM. This review systematically summarizes recent advances in four key MQC mechanisms involved in SCM: (1) phosphatase and tensin homolog-induced putative kinase 1 (PINK1)/Parkin-mediated mitophagy; (2) mitochondrial dynamics, including dynamin-related protein 1 (Drp1)/fission protein 1 (FIS1)-driven fission and optic atrophy protein 1 (OPA1)/mitofusin (MFN)-regulated fusion; (3) mitochondrial biogenesis under the regulatory control of the peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PGC-1α)/nuclear respiratory factor 1 (NRF1)/mitochondrial transcription factor A (TFAM) axis; and (4) the mitochondrial unfolded protein response (UPRmt), which maintains mitochondrial proteostasis through mediators such as C/EBP homologous protein (CHOP), YME1-like protease (YME1L), and Overlapping activity with m-AAA protease 1 (OMA1). These mechanisms have been shown to work synergistically to regulate mitochondrial clearance, renewal, and functional maintenance. Any imbalance among them can exacerbate myocardial injury. This review also emphasizes the redox crosstalk between oxidative stress and immune inflammation, with an emphasis on the pivotal contributions of NADPH oxidase 2 (NOX2), high mobility group box 1 (HMGB1), and the nucleotide-binding oligomerization domain-like receptor family pyrin domain containing 3 (NLRP3) inflammasome in vascular endothelial dysfunction and cardiac depression. In conclusion, preserving the dynamic equilibrium of MQC is crucial for preventing or reversing SCM and may present novel molecular targets and therapeutic strategies.
Myocardial infarction (MI) remains a major cause of global morbidity and mortality, with post-MI inflammation significantly exacerbating tissue damage and adverse cardiac remodeling. Recent advancements have highlighted the gut microbiota as a crucial regulator of cardiovascular health via the gut-heart axis. This review introduces a novel therapeutic perspective by exploring the potential mechanisms through which fucoidan, a sulfated polysaccharide derived from marine algae, may alleviate post-MI inflammation. Fucoidan demonstrates unique anti-inflammatory and antioxidant properties and has been shown to modulate the gut microbiota. Specifically, it enhances the abundance of beneficial gut bacteria, boosts short-chain fatty acids (SCFA) production, and reduces bacterial endotoxin translocation. Additionally, fucoidan directly inhibits inflammatory signaling pathways, decreases pro-inflammatory cytokine secretion, and protects cardiomyocytes through antioxidant effects. These multifaceted actions position fucoidan as a promising novel therapeutic strategy for mitigating post-MI inflammation and promoting cardiac recovery. Future research should focus on optimizing fucoidan extraction methods, identifying effective dosing regimens, and developing targeted delivery systems to maximize clinical efficacy.
Bone marrow failure (BMF) syndromes encompass inherited disorders, immune-mediated aplasia, and clonal myeloid neoplasms, which share overlapping biology but divergent clinical trajectories. Inflammation shapes hematopoietic stem and progenitor cell fate across this spectrum, yet its network organization, genetic conditioning, and therapeutic implications remain incompletely defined. An integrative, literature-based analysis of inflammatory cytokines, genetic contexts, and therapeutic targeting across major BMF disorders, including inherited BMF syndromes, aplastic anemia (AA), myelodysplastic syndromes (MDS), myeloproliferative neoplasms (MPNs), and myelofibrosis (MF). Gene-inflammation-outcome networks and co-occurrence maps were constructed with stratification by germline and somatic alterations. Therapeutic activity was mapped across cytokine-directed and pathway-targeted agents. Inflammatory signaling emerged as a conserved feature across all BMF categories, with TNF-α, IFN-γ, and IL-6 forming a recurrent core module, and TGF-β linking inflammatory programs to fibrotic remodeling. TP53- and JAK2-associated pathways appeared as central nodes integrating multi-cytokine inputs. Germline lesions in FANCA, TERC/TERT, and GATA2 were predominantly associated with apoptotic stem-cell attrition. In contrast, somatic drivers, including TP53, JAK2, TET2, DNMT3A, and ASXL1, were more frequently associated with clonal expansion. Multi-cytokine convergence on shared effectors was common, suggesting pathway redundancy. The therapeutic literature predominantly focused on TNF-α/IFN-γ/IL-6 pathways, with comparatively limited exploration of TGF-β, IL-8, and IL-17. These findings support a role for inflammation as a unifying, context-dependent modifier across the BMF spectrum, and highlight TP53- and JAK2-centered circuitry as potential nodes for rational combination strategies. TGF-β, IL-8, and IL-17 axes and inherited BMF entities represent underexplored opportunities for biomarker-guided cytokine-directed intervention.
Fungal esophagitis, predominantly caused by Candida species, remains the leading form of infectious esophagitis worldwide. Although historically linked to human immunodeficiency virus infection, its epidemiology has shifted over recent decades, with a growing incidence among oncology patients receiving immune checkpoint inhibitors, individuals with eosinophilic esophagitis on oral corticosteroids, and solid-organ transplant recipients. Increasing evidence highlights that disease pathogenesis extends beyond simple fungal overgrowth and involves intricate host-pathogen interactions. Pathogenic mechanisms such as adhesion, hyphal transition, and biofilm formation enhance fungal virulence, while mucosal immune dysfunction, particularly impaired T helper 17/interleukin 17 signaling and reduced antimicrobial peptide activity, predisposes to persistent infection. Antifungal resistance represents an emerging challenge, driven by efflux pump overexpression, ERG11 mutations conferring azole resistance, and mutations of FKS genes leading to reduced echinocandin susceptibility. These molecular mechanisms underscore the complexity of treatment and have important implications for antifungal selection and clinical management. While fluconazole remains the first-line agent for most cases, the increasing prevalence of non-albicans Candida species and drug resistance underscore the importance of alternative medications with different molecular targets. Agents such as the recently approved rezafungin as well as compounds such as ibrexafungerp and fosmanogepix, represent promising options for the treatment of refractory disease. This review synthesizes current advances in the molecular pathogenesis of fungal esophagitis, with particular emphasis on host-fungal interactions, antifungal resistance mechanisms, and novel treatment strategies. A deeper understanding of these processes is essential to improve management and to guide the development of innovative immunomodulatory and antifungal therapies.
BACKGROUND:Heart transplantation (HT) remains the primary treatment for end-stage heart failure, but graft rejection-including acute cellular rejection (ACR), antibody-mediated rejection (AMR), and chronic rejection such as cardiac allograft vasculopathy (CAV)-significantly impacts long-term patient outcomes. This study investigates the role of circulating plasma extracellular vesicle profiles as potential biomarkers for distinguishing between different rejection types following HT. METHODS:We enrolled 85 HT patients with post-transplant follow-up ranging from 2 to 140 months. The cohort included patients diagnosed with AMR (n = 23), ACR (n = 11), CAV (n = 20), and those without rejection (R0, n = 31). Extracellular vesicle profiles were analyzed using the MACSPlex Exosome Kit, and associated cytokine profiles were assessed using the MILLIPLEX Human Cytokine Panel A. Statistical analysis involved the Kruskal Wallis test followed by Dunn's post-hoc test for multiple comparisons and discriminant analysis. RESULTS:Circulating plasma extracellular vesicle profiles demonstrated significant differences across post-transplantation time strata and varied according to the type of transplant rejection. During the first year post-HT, the main discriminant factors were extracellular vesicles (EVs) characterized by tetraspanins (CD9+, CD63+) and platelet markers (CD62P+, CD42a+). Five years post-transplantation, significant differences emerged in patients with AMR and ACR (both compared to each other and to the CAV/R0 groups). This difference corresponded with an increase in EV markers associated with immune cell activity (CD3+, CD4+, CD49e+, CD86+, CD20+, CD14+, CD209+, CD1c+, CD29+). Levels of prominent EV subpopulations correlated with IL‑22 in CAV patients, whereas in AMR patients, they correlated with IL‑17 and IL‑25. CONCLUSIONS:These findings support the hypothesis that extracellular vesicles may participate in both direct and indirect antigen presentation and in the regulation of immune responses leading to allograft rejection after HT. Plasma extracellular vesicle profiles hold promise as non-invasive biomarkers for monitoring and differentiating rejection types in heart transplant recipients.
BACKGROUND:Calcium oxalate (CaOx) crystal retention on the renal tubular epithelium is a key step in urolithiasis. Phosphatidylserine (PS) exposure may facilitate crystal-cell adhesion, but the upstream signaling mechanisms and the relative contributions of impaired inward PS flipping versus outward PS redistribution remain unclear. MATERIALS AND METHODS:Global proteomic profiling using 2-dimensional electrophoresis and matrix-assisted laser desorption/ionization time-of-flight/time-of-flight mass spectrometry (2-DE/MALDI-TOF/TOF) in an immortalized human proximal tubular epithelial cell line (HK-2) cells exposed to calcium oxalate monohydrate (COM) identified upregulation of the calcium-sensing receptor (CaSR). HK-2 cells were treated with COM with or without the CaSR antagonist NPS2390 or the CaSR agonist gadolinium chloride (GdCl3). Bidirectional PS transport was assessed using an N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) (NBD)-labeled phosphatidylserine (NBD-PS) fluorescence-quenching assay, and surface PS exposure was measured by annexin V binding. Aminophospholipid translocase (APLT) expression, APLT-dependent inward PS transport, crystal adhesion, oxidative stress, and apoptosis-related signaling were evaluated. RESULTS:COM increased CaSR expression, enhanced surface PS exposure, and promoted crystal adhesion with concurrent oxidative stress and apoptosis-related signaling. COM induced a CaSR-sensitive defect in APLT-dependent inward PS flipping: NPS2390 partially restored inward PS transport and APLT expression, whereas GdCl3 exacerbated these changes. In contrast, COM-enhanced outward PS redistribution and externalization was largely unaffected by CaSR modulation, indicating relative CaSR insensitivity of the outward process. Consistently, CaSR activation aggravated, while CaSR inhibition attenuated, crystal adhesion and injury-related readouts. CONCLUSIONS:COM was associated with enhanced crystal-cell adhesion, CaSR activation, and a CaSR-sensitive impairment of APLT-dependent inward PS flipping, whereas enhanced outward PS redistribution appeared largely CaSR-insensitive. Pharmacologic inhibition of CaSR attenuated epithelial injury and crystal retention-related readouts, suggesting that CaSR may represent a potential therapeutic target.
Aging is one of the strongest risk factors for cancer, and its impact is particularly evident in malignancies of the reproductive system. Ovarian, endometrial, cervical, vulvar, prostate, and penile cancers are mainly diagnosed in older adults and often show different clinical and biological features compared with the same tumors in younger patients. Aging is associated with hormonal changes, immune decline, epigenetic alterations, and accumulation of DNA damage, all of which contribute to cancer development and progression. At the same time, many older patients have frailty and multiple comorbidities, which can limit the use of screening programs and invasive diagnostic procedures. This often leads to delayed diagnosis and worse outcomes. Cell-free DNA (cfDNA) is a minimally invasive biomarker that can be obtained from blood samples and provides molecular information on both tumor and host tissues. Circulating DNA reflects tumor-specific alterations but is also influenced by aging-related changes in DNA release, fragmentation, and methylation. For this reason, aging must be considered when cfDNA-based biomarkers are applied in clinical practice. In this review, we describe how aging influences the biology of reproductive system cancers and how these processes are mirrored in cfDNA profiles. We focus on the clinical use of cfDNA for cancer detection and monitoring in older and fragile patients. Special attention is given to repetitive elements in cfDNA, which are strongly affected by aging and tumor-related epigenetic changes and can be detected with high sensitivity even when the tumor fraction is low. We propose an integrative mechanistic framework in which age-related epigenetic and genomic changes influence both tumor biology and cfDNA composition, with transposable elements acting as a central link between aging and cancer.
BACKGROUND:Kir2.1 channels are responsible for membrane hyperpolarization of many cell types. While these Kir2.1 channels are known to be necessary for proper bone development and play a critical role in osteogenesis, the underlying mechanisms remain poorly understood. Here, we examined the effect of Kir2.1-mediated membrane hyperpolarization on the osteogenic differentiation of human dental follicle stem cells (hDFCs), a type of mesenchymal stem cell (MSC), and explored the underlying mechanisms. METHODS:Levels of Kir2.1 and osteogenic marker expression were evaluated by quantitative real-time polymerase chain reaction (qRT-PCR) and western blotting. Alkaline phosphatase (ALP) and Alizarin red staining were employed to evaluate ALP enzymatic activity and mineralized nodule formation, respectively. Intracellular Ca2+ levels were measured using fluorescent Ca2+ indicators with Ca2+ imaging. RESULTS:Reversal of membrane hyperpolarization via modulation of extracellular K+ concentration ([K+]e) was shown to suppress osteogenic differentiation of hDFCs, whereas the induction of membrane hyperpolarization through the opening of ATP-sensitive K+ channels had the opposite effect, enhancing hDFC osteogenesis. Kir2.1 channel expression was upregulated during the osteogenic differentiation of hDFCs. Inhibition of Kir2.1 using short hairpin RNA (shRNA) or a pharmacological inhibitor suppressed osteogenic differentiation. Hyperpolarizing the membrane by decreasing [K+]e led to the elevation of intracellular Ca2+ levels, whereas this effect was eliminated by the removal of extracellular Ca2+, Kir2.1 inhibition, or treatment with La3+, a store-operated Ca2+ channel (SOC) blocker. CONCLUSIONS:Our findings indicate that membrane hyperpolarization promotes osteogenic differentiation of hDFCs by increasing intracellular Ca2+ levels and that the Kir2.1 and SOC channels play important roles in this process.
Metabolic dysfunction-associated steatotic liver disease (MASLD) affects more than 1.66 billion people globally. This metabolic disorder shows strong connections with mitochondrial dysfunction and disrupted mitochondrial quality control (MQC) processes. Our review systematically analyzes the molecular mechanisms underlying the relationship between MQC and MASLD development. Research evidence indicates that impaired MQC regulation worsens three key pathological conditions: it intensifies liver fat accumulation, increases oxidative damage, and promotes inflammatory cell death through activating inflammatory signaling pathways, boosting reactive oxygen species generation, and triggering programmed cell death in hepatocytes. Current pharmacological interventions have shown mixed effects. Metformin, resveratrol, and empagliflozin have moderating effects on MQC homeostasis in experimental models, but their clinical application remains limited. The review further predicts two key systemic impacts of MASLD: it amplifies cardiovascular disease risk through links between organ systems, and it highlights the need to develop targeted regulation of the gut microbiota. Three major research challenges require urgent attention: first, clarifying the complex interactions between different MQC pathways; second, establishing clinical effectiveness through human trials; third, bridging the gap between laboratory findings and practical medical interventions. These priorities highlight the need for coordinated multidisciplinary research efforts to address this growing public health challenge.
BACKGROUND:Sustained activation of adipose tissue macrophages (ATMs) drives metabolic dysfunction in obesity, with the hypoxia-inducible factor-1α (HIF-1α)/interleukin-1β (IL-1β) axis being a core signaling pathway. However, specific strategies targeting this axis are still lacking. Here, we aimed to identify novel compounds capable of disrupting this pathway. METHODS:We screened a natural product library for inhibitors of IL-1β expression in macrophages. The hit compound LJ-6 was then evaluated in lipopolysaccharide (LPS)-stimulated bone marrow-derived macrophages (BMDMs) and in a physiologically relevant model using conditioned medium from obese adipose tissue. Mechanism of action was investigated via Western blotting, molecular docking, dynamics simulations, and hypoxia-response element (HRE)-luciferase reporter assays with site-directed mutagenesis. RESULTS:LJ-6 was identified as a potent inhibitor of IL-1β. It dose-dependently suppressed IL-1β in both LPS-induced and obese adipose tissue-conditioned medium-induced macrophage inflammation. Mechanistically, LJ-6 reduced HIF-1α protein stability. Molecular docking and mutagenesis studies supported an interaction involving the Tyr92 and His197 residues of HIF-1α, and suggested that this interaction is critical for its inhibitory function on the HIF-1α/IL-1β axis. CONCLUSIONS:This study identifies the natural compound LJ-6 as a novel direct inhibitor of HIF-1α. By binding to HIF-1α, LJ-6 effectively suppresses the HIF-1α/IL-1β axis in macrophages, thereby reducing IL-1β-driven inflammatory responses in cellular and ex vivo models of obesity-associated adipose tissue inflammation, and thus represents a promising lead compound for therapeutic development.
BACKGROUND:Hepatocellular carcinoma (HCC) is the third leading cause of cancer-related deaths worldwide. However, the role of small ubiquitin-like modifier 2 (SUMO2), a core member of the small ubiquitin-like modifier (SUMO) family, regarding its expression patterns and metabolism-related functions in HCC remains inadequately understood. METHODS:A multidimensional analytical framework was applied, integrating immunohistochemistry (153 HCC vs. 21 non-HCC samples), proteomics (159 paired samples), bulk transcriptomics (3240 HCC vs. 2267 non-HCC samples), single-cell RNA sequencing (RNA-seq) (10 HCC vs. 8 non-HCC samples), spatial transcriptomics, and external CRISPR/Cas9 functional genomics data. Systematic analyses included standardized mean difference (SMD), pathway enrichment, pseudotime trajectory inference, in silico knockout, cell-cell communication, metabolic flux scoring, immune infiltration, clinical correlation, drug sensitivity prediction, and molecular docking. RESULTS:At the protein level, immunohistochemistry (nuclear positivity) and external proteomic data collectively demonstrated consistent SUMO2 overexpression in HCC. Consistent upregulation was also observed at the mRNA level across large-scale cohorts. Single-cell RNA-seq and spatial transcriptomics localized SUMO2 enrichment to malignant hepatocytes and tumor-dominant regions. CRISPR-mediated SUMO2 knockout suppressed proliferation in multiple HCC cell lines. Mechanistically, high SUMO2 expression was significantly associated with metabolic reprogramming involving glycolysis/gluconeogenesis, pyruvate metabolism, and the tricarboxylic acid cycle. SUMO2-high malignant hepatocyte subpopulations exhibited enhanced activity of the macrophage migration inhibitory factor signaling axis and enhanced iron-sensor interactions. Further, the immune infiltration analysis revealed a negative correlation between SUMO2 expression and M1 macrophages and a positive correlation between follicular helper T cells and regulatory T cells. Clinically, elevated SUMO2 levels were found to be associated with adverse prognostic features. Furthermore, high SUMO2 expression was associated with increased sensitivity to dasatinib, and molecular docking simulations predicted potential binding between SUMO2 and dasatinib, with a Vina score of -8.5 kcal/mol. CONCLUSIONS:SUMO2 is aberrantly expressed at the protein, mRNA, single-cell, and spatial transcriptomic levels in HCC and is significantly associated with metabolic reprogramming and altered migration inhibitory factor (MIF)-mediated intercellular communication, suggesting its potential as a novel biomarker for diagnosis and treatment.
