OBJECTIVE:To investigate the role of early growth response 1 (EGR1) in the progression of endometrial cancer (EC) and to elucidate the molecular mechanism by which EGR1 regulates ferroptosis, with a focus on NADPH oxidase 4 (NOX4) as a downstream mediator. METHODS:An EC xenograft model was established in nude mice. EGR1 was overexpressed in HEC-1B cells, and cell proliferation and migration were assessed using CCK-8, colony formation, and Transwell assays. Ferroptosis-related indicators (Fe2+, MDA, GSH, and lipid ROS) and related proteins (GPX4, SLC7A11, and IREB2) were measured. The ferroptosis inhibitor Fer-1 was used for rescue experiments. Gene and protein expression were analyzed via RT-qPCR, western blotting, and immunohistochemistry. The transcriptional regulation of NOX4 by EGR1 was validated using chromatin immunoprecipitation (ChIP) and dual-luciferase reporter assays. RESULTS:EGR1 expression was downregulated in EC. Overexpression of EGR1 inhibited the proliferation and migration of HEC-1B cells and suppressed tumor growth in vivo. EGR1 overexpression induced ferroptosis, as evidenced by increased levels of Fe2+, MDA, and lipid ROS; decreased GSH levels; downregulation of GPX4 and SLC7A11; and upregulation of IREB2. These effects were reversed by Fer-1. Mechanistically, EGR1 directly bound to the NOX4 promoter and increased its transcription, leading to increased ROS production and thereby facilitating ferroptosis. CONCLUSION:EGR1 inhibits the progression of EC at least in part through transcriptionally activating NOX4, which promotes ROS accumulation and contributes to ferroptosis induction. This study provides a theoretical basis for targeting EGR1 as a potential therapeutic strategy for treating EC.
This study aimed to investigate the role of Src homology collagen-binding protein 1 (SHCBP1) in the progression and cisplatin resistance of laryngeal squamous cell carcinoma (LSCC), and to elucidate the underlying regulatory mechanism. LSCC cell lines were used to assess SHCBP1 expression by western blotting and quantitative real-time PCR (qRT-PCR). SHCBP1 knockdown was achieved by siRNA transfection, and its effects on cell proliferation, migration, and cisplatin sensitivity were evaluated through cell viability assays, wound healing assays, and flow cytometry. The transcriptional regulation of SHCBP1 by POU class 2 homeobox 1 (POU2F1) was investigated using dual-luciferase reporter assays and chromatin immunoprecipitation (ChIP) analysis. In vivo, tumor xenograft models were established to determine the impact of SHCBP1 knockdown on tumor growth and cisplatin responsiveness. SHCBP1 was highly expressed in LSCC cells, and its knockdown significantly inhibited LSCC cell proliferation and migration. Moreover, SHCBP1 silencing enhanced the sensitivity of LSCC cells to cisplatin treatment. Mechanistically, POU2F1 was shown to activate SHCBP1 transcription in LSCC cells. In vivo experiments further confirmed that SHCBP1 knockdown suppressed tumor growth and metastasis and increased cisplatin sensitivity. Overall, SHCBP1 contributes to LSCC cisplatin resistance and progression by promoting POU2F1-mediated transcriptional activation.
BACKGROUND:Acute lung injury (ALI) is a critical condition characterized by severe inflammatory responses. High-density lipoprotein (HDL) has been demonstrated to possess anti-inflammatory properties, yet its mechanistic role in ALI through the SIRT1/NF-κB/NLRP3 signaling axis remains to be elucidated. OBJECTIVE:This study aimed to investigate the role of HDL in ALI and to elucidate its underlying mechanisms, with a focus on modulation of the SIRT1/NF-κB/NLRP3 signaling pathway. METHODS:BEAS-2B cells were exposed to lipopolysaccharide (LPS) and treated with HDL (20-200 μg/mL). Cellular viability was assessed using CCK-8. In vivo, LPS-induced ALI mice received HDL treatment with or without the SIRT1 inhibitor EX-527. Lung pathology and edema were evaluated via H&E staining and wet-to-dry (W/D) ratios. Inflammatory markers and myeloperoxidase (MPO) activity were measured using qPCR and ELISA, while western blotting analyzed SIRT1, NF-κB, and NLRP3 inflammasome components. RESULTS:HDL at concentrations ≤100 μg/mL effectively mitigated LPS-induced cytotoxicity and inflammation in BEAS-2B cells (p<0.01). This protective effect was dose-dependent, evidenced by reductions in the expression of IL-6 (p<0.01), IL-1β, TNF-α (p<0.05), NLRP3, and caspase-1, alongside restoration of SIRT1 activity suppressed by LPS (p<0.01). In vivo, HDL (40 mg/kg) significantly improved pulmonary histopathology and reduced serum pro-inflammatory cytokine levels and MPO activity in ALI mice (p<0.01). Importantly, co-treatment with EX-527 abolished HDL´s protective effects, underscoring the pivotal role of SIRT1. Mechanistically, HDL upregulated SIRT1, which subsequently inhibited NF-κB signaling and suppressed NLRP3 inflammasome activation in a SIRT1-dependent manner (p<0.01). This sequential inhibition disrupted the NF-κB-driven cytokine amplification loop. CONCLUSION:HDL confers robust protection against LPS-induced ALI by upregulating SIRT1, thereby suppressing the NF-κB/NLRP3/IL-1β inflammasome signaling axis. These findings highlight the HDL-SIRT1 axis as a potential therapeutic target for ALI, supporting the development of HDL mimetics or SIRT1 activators.
BACKGROUND:This study aimed to evaluate the association between pretreatment immunohistochemical expression of hypoxia-inducible factor 1 alpha (HIF-1α) and excision repair cross-complementation group 1 (ERCC1) and clinical outcomes in patients with locally advanced cervical cancer (LACC) treated with definitive concurrent radiochemotherapy (cRCT). METHODS:Eighty-seven patients with histologically confirmed LACC who received definitive cRCT between 2005 and 2015 were retrospectively analyzed. Immunohistochemical staining for HIF-1α and ERCC1 was performed on pretreatment biopsy specimens, and H-scores were calculated semi-quantitatively. Receiver operating characteristic (ROC) curve analysis determined optimal cut-off values: ≤100 and >100 for HIF-1α, and ≤130 and >130 for ERCC1. Overall survival (OS), local recurrence-free survival (LRFS), and distant metastasis-free survival (DMFS) were estimated using the Kaplan-Meier method, and prognostic variables were analyzed with Cox regression models. RESULTS:The median age was 57 years (range: 37-85). The five-year OS, LRFS, and DMFS rates were 57.8%, 69.7%, and 69.1%, respectively. In multivariate analysis, incomplete treatment response (p<0.001) and high neutrophil-to-lymphocyte ratio (p=0.042) were independent predictors of poor OS. For LRFS, tumor size >4 cm (p=0.048), incomplete response (p<0.001), and high ERCC1 expression (p=0.032) were identified as unfavorable prognostic factors. High ERCC1 expression also correlated with increased late toxicity (p=0.038). HIF-1α expression showed no significant association with any survival outcome. CONCLUSIONS:High ERCC1 expression was an independent prognostic indicator for poor LRFS, suggesting its potential as a biomarker of radioresistance. In patients with elevated ERCC1, alternative non-platinum agents may be considered. HIF-1α had no predictive or prognostic value in this cohort.
BACKGROUND:Acute lung injury (ALI) induced by lipopolysaccharide (LPS) involves severe inflammation and epithelial damage, necessitating effective therapeutic strategies. This study investigated the protective effects and mechanisms of salvianic acid A sodium (SAAS) against LPS-induced cytotoxicity and ALI. METHODS:In vitro cytoprotection and anti-inflammatory effects of SAAS (0-60 μM) were assessed in LPS-challenged alveolar type II (AT2) epithelial cells using CCK-8 assays and quantification of inflammatory mediators (NLRP3, ASC, IL-1β, IL-6, TNF-α). In vivo efficacy was evaluated in an LPS-induced murine ALI model treated with SAAS via histopathology, lung wet/dry weight ratio, myeloperoxidase (MPO) activity, serum/lung cytokine levels (ELISA), and immunohistochemistry (IHC). Mechanistic insights were gained through western blot and qPCR analysis of HMGB1 signaling components. RESULTS:SAAS (≤60 μM) exhibited no intrinsic cytotoxicity but dose-dependently attenuated LPS-induced AT2 cell death. It significantly suppressed LPS-triggered expression of NLRP3 inflammasome components (NLRP3, ASC), caspase-1, and pro-inflammatory cytokines (IL-1β, IL-6, TNF-α) in vitro. In the ALI model, SAAS administration ameliorated histopathological damage, attenuated pulmonary edema, suppressed MPO activity, and reduced systemic and pulmonary levels of TNF-α, IL-6, IL-18, and IL-1β. Mechanistically, SAAS inhibited LPS-induced HMGB1 upregulation, acetylation, and nuclear-to-cytoplasmic translocation. Consequently, it suppressed downstream NF-κB activation and NLRP3 inflammasome assembly/activation. CONCLUSION:SAAS confers significant protection against LPS-induced ALI by targeting the HMGB1 signaling axis. It inhibits HMGB1 expression, acetylation, and cytoplasmic translocation, thereby disrupting NF-κB-mediated cytokine production and NLRP3 inflammasome activation. These findings identify SAAS as a promising multi-target therapeutic agent for inflammatory lung injury.
Satellite cells (SCs) are recognized as the resident stem cells of adult skeletal muscles, essential for post-natal skeletal muscle growth and regeneration following focal myotrauma. In healthy muscle, SCs are quiescent, but in response to damage or growth signals, they become activated to proliferate and differentiate to form new myofibers. A small population self-renews to replenish the basal pool for future demands. The ability of SCs to precisely balance quiescence, self-renewal, and myogenic commitment/differentiation is essential for ensuring long-term muscle homeostasis and tissue maintenance. Their state and functionality are strictly regulated by different intrinsic and extrinsic cues, the latter deriving from the microenvironment in which SCs reside, known as the niche. The niche is a dynamic compartment where extracellular matrix components, soluble factors, mechanical stimuli, and multiple interacting cell populations modulate the morphological, molecular, and electrophysiological properties of SCs. This review provides an updated overview of the morpho-functional features of SCs and of non-myogenic stromal interstitial cells, highlighting their reciprocal crosstalk within the regenerative niche. Such stromal cells play a dual role, acting as "good" or "bad" cells: while functioning as nursing cells for SCs during muscle repair/regeneration via juxtracrine and paracrine interactions, their excessive accumulation and adoption of a fibrotic/fat phenotype may lead to aberrant tissue repair, compromising muscle function. A deeper understanding of SC biology and of collaborative spatiotemporal cell interactions in the healthy, damaged, and regenerative niche is essential to identify potential novel targets and to better address interventions for maintaining, restoring, or enhancing muscle regeneration capacity and mitigating the deleterious effects of extended, severe, or pathological muscle damage.
The thymus is a key lymphoid organ responsible for T-cell development, yet information on its postnatal dynamics in the dromedary camel remains limited. In this study, we examined age-related morphological and histological changes in the camel thymus across four age groups (up to 1 year, 1-3 years, 3-5 years, and over 5 years) using fixed and fresh anatomical samples and stained paraffin-embedded sections. The thymus consisted of cervical and thoracic parts, with the thoracic thymus reaching its maximal size shortly before or at one year of age and achieving its greatest weight at one year, followed by a progressive decline thereafter in size, accompanied by increasing fat infiltration. Histologically, advancing age was associated with thickening of the interlobular septa, reduced lobular diameter, and the progressive replacement of parenchymal tissue with fibrous and adipose components, and keratinization or degeneration of Hassall's corpuscles. While the number of Hassall's corpuscles peaked in 1-3-year-old camels and declined thereafter, their thickness increased progressively across age groups. Collectively, these findings indicate an early onset of thymic maturation followed by gradual involution in the dromedary camel, providing valuable insight into the timeline and structural basis of immunosenescence in this species.
OBJECTIVE:To investigate the anti-fibrotic effects of Isofraxidin (IF) on liver fibrosis and its underlying mechanisms. METHODS:C57BL/6J mice (bred by The Jackson Laboratory) were induced with liver fibrosis using CCl4 and treated with different doses of IF (20 mg/kg and 40 mg/kg) via gavage. Colchicine served as the positive control. Liver fibrosis markers (α-smooth muscle actin,α-SMA; Fibronectin, FN; Collagen type I, Col-I) were assessed by histological staining, immunohistochemistry, and western blotting. Network pharmacology and molecular docking were adopted to conjecture underlying targets and signaling pathways of IF, followed by experimental validation. RESULTS:IF at various doses improved liver function and exhibited dose-dependent anti-fibrotic effects. Network pharmacology identified Protein kinase Kinase B (PKB, Akt), Proto-oncogene tyrosine-protein kinase (Src), Epidermal growth factor receptor (EGFR), Heat Shock Protein 90 Alpha Family Class A Member 1 (HSP90AA1), Glycogen Synthase Kinase 3 beta (GSK3β), and Monoamine Oxidase B (MAOB) as the core targets. Molecular docking showed good binding affinity between IF and these targets. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses indicated that the Nuclear Factor kappa-light-chain-enhancer of activated B cells (NF-κB) signaling pathway is a major pathway involved in IF's anti-fibrotic effects. Experimental validation confirmed that IF inhibited the expression of inflammatory factors (Tumor Necrosis Factor-alpha, TNF-α; Interleukin-1 beta, IL-1β; Interleukin-10, IL-10) and the phosphorylation of NF-κB and Inhibitor of Nuclear Factor Kappa B Alpha (IκBα). CONCLUSION:IF effectively alleviated CCl4-induced liver fibrosis in mice by modulating key targets (AkT, Src, EGFR, HSP90AA1, GSK3B, and MAOB) and the NF-κB signaling pathway.
BACKGROUND:Melanoma represents a highly aggressive form of skin cancer characterized by significant invasiveness and immune evasion capabilities. Although heme oxygenase-1 (HMOX1) participates in cellular stress responses and ferroptosis, its precise functional roles in melanoma pathogenesis remain incompletely defined. METHODS:HMOX1 expression levels in melanoma tissues and cell lines were evaluated using immunohistochemistry (IHC), quantitative real-time PCR (qRT-PCR), and western blotting. HMOX1 function was investigated through short hairpin RNA (shRNA)-mediated knockdown, followed by assessment of cellular proliferation, migration, invasion, and ferroptosis sensitivity. Forkhead box O3 (FOXO3)-mediated transcriptional regulation of HMOX1 was experimentally validated using dual-luciferase reporter assays. The impact of HMOX1 expression in melanoma cells on CD8+ T cells was examined through co-culture experiments with peripheral blood mononuclear cells (PBMCs). The in vivo effect of HMOX1 knockdown was evaluated using a mouse subcutaneous xenograft model. RESULTS:HMOX1 expression was significantly elevated in melanoma compared with normal controls and strongly correlated with poor prognosis. HMOX1 knockdown effectively suppressed melanoma cell proliferation, migration, and invasion, while simultaneously inducing ferroptosis, characterized by increased intracellular ferrous iron (Fe2+) and reactive oxygen species (ROS) levels, along with decreased glutathione peroxidase 4 (GPX4) expression. Additionally, HMOX1 depletion attenuated immune escape mechanisms, evidenced by reduced programmed death-ligand 1 (PD-L1) expression, promoted CD8+ T cell expansion in co-culture systems, and elevated tumor necrosis factor-alpha (TNF-α) and interferon-gamma (IFN-γ) secretion. Mechanistically, the transcription factor FOXO3 was identified as a transcriptional activator of HMOX1, thus establishing a FOXO3/HMOX1 regulatory axis that drives melanoma progression. In vivo xenograft experiments corroborated these findings, demonstrating significant tumor growth inhibition upon HMOX1 silencing. CONCLUSION:FOXO3-mediated upregulation of HMOX1 promotes melanoma cell proliferation, invasion, immune escape, and ferroptosis resistance. The FOXO3/HMOX1 axis represents a potential therapeutic target for melanoma treatment.
CD44, a cell-surface glycoprotein involved in cell-cell interactions and adhesion, has been implicated in tumorigenesis and cancer stem cell (CSC) maintenance in various human malignancies. While its expression has been investigated in oral squamous cell carcinoma (OSCC), further evaluation from diverse molecular perspectives is warranted to fully understand its clinical and molecular relevance. Therefore, this study aimed to immunohistochemically evaluate CD44 expression and its correlation with clinicopathological factors and molecular markers involved in cellular signaling (EGFR, STAT3) and lipid metabolism (FAS, SREBP1, E-FABP) in 50 cases of OSCC. Positive CD44 immunoreactivity was observed in 49 of 50 cases (98.0%). While CD44 expression was not statistically correlated with clinicopathological factors such as tumor differentiation, pT stage, or pTNM stage in this cohort, significant positive correlations were found between CD44 expression and EGFR (p=0.021), STAT3 (p=0.008), and FAS (p=0.010) expression. No significant correlations were observed with SREBP1 (p=0.057) or E-FABP (p=0.058). These findings suggest that CD44 expression is statistically associated with EGFR, STAT3, and FAS expression in OSCC. Although a direct correlation with clinicopathological aggressiveness was not evident in this study, the co-expression of CD44 with these molecular markers provides a basis for the hypothesis that CSC properties may be interrelated with signal transduction and lipid metabolic reprogramming in oral cancer cells.
BACKGROUND:Although betulinic acid (BA) possesses diverse pharmacological properties, its cardioprotective potential in sepsis-associated myocardial damage remains undefined. This investigation explores BA's therapeutic potential and underlying mechanisms against lipopolysaccharide (LPS)-induced septic cardiomyopathy in murine models. METHODS:Experimental sepsis was induced via intraperitoneal LPS administration (10 mg/kg), with subjects randomized into six cohorts: normal controls, LPS model group, positive control (imipenem 120 mg/kg), and BA treatment arms (15/30/60 mg/kg oral gavage). Pharmacological interventions preceded the LPS challenge by seven days. Comprehensive cardiac evaluation encompassed echocardiographic parameters, ELISA quantification, histopathological assessment (H&E), with complementary TUNEL and DCFH-DA staining for pyroptotic quantification and oxidative stress evaluation. Immunoblotting analyzed the TLR4/MyD88/NF-κB cascade, MAPK phosphorylation states, and NLRP3 inflammasome activation. RESULTS:BA administration demonstrated dose-dependent cardioprotection, evidenced by normalized ejection fraction (echocardiography), attenuated serum biomarkers of myocardial injury (LDH, CK-MB, and cTnI), and preserved histoarchitecture (H&E). Mechanistically, BA suppressed proinflammatory cytokine secretion (TNF-α, IL-1β, IL-6) through dual inhibition of TLR4-mediated NF-κB nuclear translocation and MAPK (p38/JNK) phosphorylation cascades. In vitro validation using H9c2 cardiomyoblasts (12.5-50 μM BA) confirmed NLRP3 inflammasome downregulation, evidenced by reduced GSDMD cleavage, caspase-1 activation, and interleukin maturation. CONCLUSION:These findings substantiate BA's myocardial preservation through coordinated modulation of the TLR4/NF-κB/MAPK axis and NLRP3 inflammasome activation, proposing BA as a novel multi-target therapeutic candidate for sepsis-induced cardiac dysfunction.
Extension loss is a prevalent and clinically debilitating complication following anterior cruciate ligament reconstruction (ACLR). Joint immobilization is widely recognized as a primary etiology of range of motion (ROM) restriction across various musculoskeletal pathologies. Even following ACLR, diminished joint excursion during activity likely contributes to the development of extension loss, and postoperative joint immobilization exacerbates extension loss by facilitating intra-articular adhesion formation. Although early mobilization strategies, such as continuous passive motion, have traditionally been the cornerstone of postoperative rehabilitation aimed at counteracting the deleterious effects of immobilization, this approach often fails to fully prevent or resolve extension loss. Translational research indicates that post-ACLR extension loss progresses through distinct biological stages. During the acute postoperative period, termed the active arthrofibrosis phase, intra-articular hemorrhage and subsequent inflammation trigger fibrotic responses, resulting in adhesion formation and joint capsule thickening. In the later residual arthrofibrosis phase, persistent fibrosis may lead to persistent extension loss despite resolution of inflammation. Crucially, the efficacy of therapeutic interventions is highly phase-dependent. Aggressive exercise during the active arthrofibrosis phase may paradoxically exacerbate inflammation and fibrosis, whereas mechanical loading during the residual arthrofibrosis phase may facilitate ROM recovery. In addition, adjunctive pharmacological and physical modalities targeting inflammation may mitigate subsequent fibrotic progression. This narrative review synthesizes clinical and basic research evidence to elucidate the pathophysiological mechanisms underlying extension loss after ACLR and proposes a phase-dependent framework for its management. A paradigm shift toward biologically informed, timing-specific interventions is essential to minimize the risk of persistent extension loss.
OBJECTIVES:Knee osteoarthritis (KOA) is a progressive degenerative condition prevalent in older adults. ShenQiWan (SQW) has shown notable clinical efficacy in treating KOA. However, the role and mechanism of SQW in KOA are still poorly understood. METHODS:Here, a destabilization medial meniscus (DMM)-induced KOA mouse model and LPS-induced chondrocytes were established to evaluate the protective effect of SQW in treating KOA. The impact of SQW on KOA cartilage and subchondral bone was assessed via micro-CT, histological examination, and immunohistochemical analysis. Simultaneously, LPS-stimulated KOA chondrocytes were treated with SQW to assess the proliferation, apoptosis, and inflammatory responses via CCK-8, ELISA, flow cytometry, RT-qPCR, and western Blot. NLRP3 overexpression was performed in chondrocytes to explore the role of SQW-mediated serum on the apoptosis and inflammation in LPS-induced chondrocytes by regulating the NF-κB/NLRP3 pathway. RESULTS:SQW effectively improved gait and reduced pain-related behaviors in KOA mice. Micro-CT and histological analyses revealed that SQW alleviated subchondral bone loss and cartilage destruction, improved joint pathology. Molecular analyses revealed that SQW inhibited chondrocyte apoptosis and reduced the inflammatory response. Mechanistically, SQW down-regulated the expression of NLRP3 inflammasome-related proteins and inhibited the phosphorylation of NF-κB pathway components. In vitro, SQW protected LPS-induced chondrocytes from injury, improving viability and reducing apoptosis, while NLRP3 overexpression reversed these effects, confirming SQW acts via the NLRP3/NF-κB pathway. CONCLUSIONS:SQW effectively reduces KOA symptoms by blocking the NF-κB/NLRP3 pathway, thereby lessening cartilage damage, inflammation, and cell death, while improving mobility and pain responses, highlighting its potential as a therapeutic agent for KOA.
BACKGROUND:Lung adenocarcinoma (LUAD) is a kind of lung cancer characterized by cancer stem cell (CSC) properties. Cell division cycle 6 (CDC6), a cell cycle regulator, is significantly associated with tumor progression, but its role in LUAD stemness remains unclear. METHODS:CDC6 expression levels were analyzed using RNA-seq data from the TCGA database and clinical samples. Cell sphere formation assays, immunofluorescence, and western blot were performed to evaluate the impact of CDC6 on LUAD stemness. Lactate production and the extracellular acidification rate (ECAR) were measured to assess glycolysis levels. The hTFtarget database was used to screen potential upstream regulators, and the regulatory relationship between transcription factor AP-2α (TFAP2A) and CDC6 was validated by luciferase reporter assays and ChIP-qPCR. A xenograft tumor model was established to examine the result of CDC6 knockdown on LUAD tumor progression. RESULTS:The results indicated that CDC6 expression was markedly elevated in LUAD tissues compared with adjacent normal tissues, and high CDC6 expression was associated with poorer overall survival. In vitro experiments confirmed that CDC6 enhanced LUAD stemness by activating glycolysis. Mechanistic studies revealed that TFAP2A directly bound to the CDC6 promoter region, regulating its transcription and expression, thereby promoting glycolysis and LUAD stemness. In vivo experiments further demonstrated that knockdown of CDC6 in LUAD cells suppressed tumor growth. CONCLUSION:This study clarifies the molecular mechanism through which TFAP2A modulates CDC6-mediated glycolysis to enhance LUAD stemness, implying that targeting the TFAP2A/CDC6 axis could serve as a potential therapeutic approach for LUAD.
The metabolic reprogramming known as the Warburg effect, in which cancer cells preferentially convert glucose to lactate even in the presence of oxygen, is a defining hallmark of malignant transformation. Rather than reflecting defective mitochondria or a simple adaptation to hypoxia, aerobic glycolysis now emerges as a central hub that integrates glycolysis, the tricarboxylic acid cycle, the pentose phosphate pathway, and lactate metabolism to sustain the bioenergetic, biosynthetic, and redox demands of cancer cells. Enhanced glycolytic flux supports rapid ATP generation, nucleotide and lipid biosynthesis, NADPH production, and acetyl-CoA-dependent epigenetic regulation, thereby promoting DNA repair, antioxidant defense, chromatin remodeling, and proliferation. Beyond these metabolic outputs, many glycolytic enzymes, such as hexokinase 2, phosphoglycerate mutase 1, pyruvate kinase M2, phosphoglucose isomerase/autocrine motility factor, and aldolase A, exert non-enzymatic "moonlighting" functions that regulate apoptosis, checkpoint signaling, transcriptional programs, cytoskeletal remodeling, and epithelial-mesenchymal transition. Through this combination of enzymatic and non-enzymatic activities, the Warburg effect reinforces multiple dimensions of therapeutic resistance, including ATP-dependent drug efflux, resistance to DNA-damaging therapies, immune evasion, and invasive, EMT-associated phenotypes. In this review, we summarize current evidence linking aerobic glycolysis, lactate metabolism, and the pentose phosphate pathway to resistance against chemotherapy, radiotherapy, targeted therapy, and immunotherapy, with a particular focus on the non-metabolic functions of glycolytic enzymes. We further discuss how targeting glucose uptake, glycolysis, lactate transport, or pentose phosphate pathway flux, alone or in combination with conventional therapies and immune checkpoint blockade, may provide metabolism-informed strategies to overcome drug resistance in cancer.
OBJECTIVES:Acupuncture, a core component of traditional Chinese medicine, is widely used for treating ischemic stroke; however, its neuroprotective mechanisms remain unclear. This study investigated the pro-angiogenic and neuroprotective effects of acupuncture following ischemic stroke and examined the potential involvement of DLL4/Notch1 signaling in these processes. METHODS:A middle cerebral artery occlusion (MCAO) rat model was established to assess acupuncture's therapeutic effects. Body weight, survival, and motor function (Bederson score) were evaluated, infarct size was determined by TTC staining, and CD31 expression was examined by immunohistochemistry. Western blot and RT-qPCR were used to measure HIF-1α, DLL4, Notch1, and VEGF expression. In vitro, acupuncture serum was evaluated as a modulatory factor in BMSC endothelial differentiation and was found to enhance cellular responsiveness to VEGFA165 and bFGF rather than acting as an independent angiogenic inducer, while DLL4 overexpression and knockdown were used to clarify its role in angiogenesis. In PC12 OGD/R cells, DLL4 plasmid transfection assessed viability (CCK-8), apoptosis (Hoechst and Annexin V/PI), and network-like phenotypes, along with Notch1/Hes1/Hes5, HIF-1α, and VEGF pathway analysis. RESULTS:Acupuncture improved survival, motor function, and body weight, reduced infarct size, and increased microvascular density, associated with HIF-1α downregulation and DLL4, Notch1, and VEGF upregulation. Acupuncture serum alone had limited instructive capacity for endothelial differentiation, but acted permissively to enhance BMSC responsiveness to VEGFA165 and bFGF, resulting in improved differentiation-associated marker expression and tube formation. DLL4 overexpression enhanced vascular integrity, while knockdown reversed these effects. In PC12 OGD/R cells, DLL4 overexpression reduced apoptosis, improved viability, and enhanced network-like phenotypes, accompanied by increased Notch1/Hes1/Hes5 and VEGF expression and reduced HIF-1α expression. CONCLUSIONS:Acupuncture facilitates angiogenesis and neurofunctional recovery after ischemic stroke, with DLL4/Notch1-associated regulatory responses potentially contributing to these effects. DLL4 upregulation may further enhance angiogenic and neuroprotective outcomes, suggesting that DLL4 may represent a potential therapeutic target in acupuncture-mediated post-stroke repair.
Glial cells, particularly microglia and astrocytes, orchestrate the neuroinflammatory response in the central nervous system (CNS) through sequential phenotypic changes encompassing inflammatory activation, migration, phagocytosis, and tissue remodeling. Kinases serve as central regulators of these processes, yet systematic investigations of kinase signaling networks in brain glia have been remarkably limited compared to oncology or peripheral immunology. To address this gap, we developed a multiplexed kinome-wide small interfering RNA screening platform that simultaneously interrogates 623 kinases across four distinct neuroinflammatory glial phenotypes. This approach identified distinct kinase signaling networks governing glial activation, migration, phagocytosis, and cell survival, and revealed that the same signaling pathway can exert opposing effects depending on the phenotype examined. Most unexpectedly, T-cell receptor signaling components were enriched among kinases promoting glial activation, leading to the identification of interleukin-2-inducible T-cell kinase (ITK)-a canonical adaptive immune kinase-as a potent regulator of microglial inflammatory activation. Subsequent validation demonstrated that microglial ITK drives neuroinflammation and behavioral deficits in a traumatic brain injury model through the Vav1-PLC-γ-NF-κB axis, with confirmatory evidence from brain tissues of patients with chronic traumatic encephalopathy and human microglial cells. Multiple intracellular kinase cascades-including the TLR-TAK1-IKK, MAPK, PI3K-Akt-mTOR, JAK-STAT, and SYK-BTK pathways-converge on key transcription factors such as NF-κB, AP-1, STAT1/3, and NFAT to shape glial inflammatory gene expression, and extensive cross-talk between these cascades adds further complexity to the neuroinflammatory response. Viewed alongside the emergence of Bruton's tyrosine kinase as a therapeutic target in microglia-driven multiple sclerosis, these findings suggest that the Tec kinase family constitutes a previously unrecognized signaling hub in brain innate immunity. This review discusses how systematic kinome screening reshapes our understanding of glial biology and highlights the translational potential of microglial ITK as a disease-relevant therapeutic target.
Marrow adipose tissue (MAT) is no longer considered a passive filler but a dynamic component of the bone marrow niche that actively regulates bone metabolism. This narrative review focuses on the dynamics of MAT accumulation following anterior cruciate ligament (ACL) reconstruction. Experimental studies, including rat models, demonstrate that ACL reconstruction induces MAT accumulation, which is further promoted by postoperative joint immobilization or non-weight-bearing. While MAT accumulation induced by immobilization or non-weight-bearing in intact joints is reversible upon remobilization or reloading, MAT accumulation induced by postoperative immobilization or non-weight-bearing following ACL reconstruction fails to recover even after subsequent remobilization or reloading. These findings suggest that the recovery mechanism observed in intact joints is disrupted in the post-ACL reconstruction environment, leading to potentially irreversible negative effects. In addition, this review highlights that the pathophysiology of surgery-induced MAT accumulation is distinct from pharmacological influences. For example, corticosteroid-induced MAT accumulation early after ACL reconstruction is reversible following treatment discontinuation, indicating that the reversibility of MAT accumulation is fundamentally dependent on the initiating stimulus. Furthermore, sex-related dimorphism is evident, with estrogen likely exerting a protective effect that delays MAT accumulation following ACL reconstruction in females. Given the potential contribution of aberrant MAT accumulation to osteoarthritis pathogenesis, future research should prioritize elucidating the biological crosstalk between MAT and non-osseous tissues, particularly articular cartilage. Clarifying the regulatory mechanisms underlying post-surgical MAT accumulation will be essential for developing effective preventive and therapeutic strategies aimed at preserving joint integrity and improving long-term patient outcomes.
BACKGROUND:Wild-type (WT) gastrointestinal stromal tumors (GISTs) are a rare and molecularly heterogeneous subset of GISTs lacking KIT and PDGFRA mutations. These tumors pose significant diagnostic and therapeutic challenges due to resistance to standard tyrosine kinase inhibitors and a wide spectrum of genetic drivers. METHODS:A total of 156 GIST patients were retrospectively reviewed, among whom 19 WT GIST cases were identified. Immunohistochemistry, Sanger sequencing, and targeted next-generation sequencing (NGS) covering 39 cancer-related genes were performed to evaluate clinicopathological and molecular characteristics. Variants were annotated and filtered using public mutation databases. RESULTS:Among the 19 WT GISTs, 8 (42.1%) harbored additional genetic alterations, including FLT3 (p.R973X), CDH1 (p.D786N), and TERT promoter alterations, along with copy number gain in CCND1 and copy number loss in RB1. SDHA and SDHD mutations were found in SDH-deficient tumors. Immunohistochemical analysis revealed a subset of SDHA-mutated tumors with retained SDHA protein expression. Kaplan-Meier analysis showed a difference in progression-free survival (PFS) between WT and non-WT GISTs (p=0.001); however, this finding should be interpreted with caution due to the small sample size and lack of adjusted analysis. CONCLUSION:WT GISTs exhibit considerable molecular heterogeneity with novel or rare mutations of uncertain significance. Targeted NGS enables the detection of clinically relevant alterations that may guide future diagnostic and therapeutic strategies. Our findings emphasize the importance of targeted molecular profiling and raise the potential for personalized treatment in this challenging subset of GISTs.
Broncho-pulmonary club (Clara) cells (Cc) play a protective role in the lung due to the secretion of the CC16 protein. Their normal and pathological morphology and function have long been well characterized by histologists and histopathologists. In recent years, human prostatic Cc have become the focus of intensified research. A three-dimensional cellular atlas of normal and pathologic human and experimental prostates has been developed using the most advanced research methods (single-cell RNA sequencing, RNA in situ). This review provides a comprehensive synthesis of current knowledge regarding prostatic Cc and Cc-phenotype conversion across prostate disease. Prostatic Cc possesses anti-inflammatory, regenerative, and immunoregulatory potential, and exhibits features like those of their pulmonary counterparts. Their ontogenesis, localization across different prostatic zones, gene expression profile, and histo-functional morphology are discussed and illustrated immunohistochemically using original material from the authors. Pathophysiological and morphogenetic considerations are also presented.
