Ion channels are fundamental to neuronal excitability, synaptic transmission, and the coordinated development of brain circuits. Disruptions in their function—collectively termed ion channelopathies—have emerged as central mechanisms underlying a broad spectrum of neurodevelopmental disorders (NDDs), including epilepsy, autism spectrum disorder, and intellectual disability. This review synthesizes current knowledge on the physiological roles of voltage-gated and ligand-gated ion channels during brain development and elucidates how their genetic and functional dysregulation contributes to disease pathogenesis. We examine key channel families, such as sodium, potassium, calcium, and glutamate/(gamma-aminobutyric acid) GABA receptors, as well as mechanosensitive and polymodal channels including transient receptor potential and Piezo channels, highlighting the molecular mechanisms, pathogenic variants, and circuit-level consequences of their dysfunction. Emerging therapeutic strategies are discussed, spanning subtype-specific small molecule modulators, antisense oligonucleotides, CRISPR-based genome editing, and patient-derived organoid models for precision medicine and drug screening. We also address significant challenges in the field, including cellular heterogeneity, developmental timing, and translational model fidelity. Together, these advances underscore a rapidly evolving landscape in which precision neurogenetics and integrative platforms hold promise for transforming the diagnosis and treatment of ion channel-related neurodevelopmental disorders.
Hepatobiliary malignancies remain a major clinical challenge because they are highly aggressive and resistant to therapy. In eukaryotes, N6-methyladenosine (m6A), the most prevalent internal RNA modification, regulates post-transcriptional gene expression. Insulin-like growth factor 2 mRNA-binding proteins (IGF2BP1/2/3) act as pivotal m6A readers, stabilizing coding and non-coding RNAs to modulate cancer-related signaling networks. In hepatobiliary cancers, dysregulated IGF2BP expression is associated with proliferation, metastasis, metabolic adaptation, and immune evasion, underscoring its potential as a biomarker and therapeutic targets. This review provides a comprehensive overview of IGF2BP-mediated regulatory mechanisms and explores their translational potential in precision diagnostics and targeted interventions.
Achondroplasia (ACH), the predominant inherited form of disproportionate short stature, results from specific genetic alterations in fibroblast growth factor receptor 3 (FGFR3). N6-methyladenosine (m6A) modification is reported to modulate mRNA stability and translation. The present investigation systematically explored the epigenetic regulatory function of METTL16, an m6A RNA methyltransferase, within the pathophysiological framework of ACH.
We generated an ACH mouse model via Fgfr3380R (Fgfr3ach) gene mutation. Primary chondrocytes were isolated from newborn mice and stimulated with IL-1β to induce cell death. Proximal tibia tissues were collected and analyzed with HE staining, toluidine blue staining, safranin O staining, and immunohistochemical (IHC) analysis. Bone structure was analyzed by measuring bone mineral density (BMD), ratio of bone volume to total tissue volume (BV/TV), trabecular number (TbN), and trabecular thickness (TbTh). Cell viability and proliferation were assessed using the Cell Counting Kit-8 (CCK-8) and colony formation assays. The levels of iron (Fe2+), malondialdehyde (MDA), and glutathione (GSH) were measured to assess ferroptosis. Protein and RNA levels were measured by western blotting and quantitative real-time PCR (qPCR) assay, respectively, while the m6A modification level was assessed by m6A mRNA immunoprecipitation (IP).
METTL16 improved bone chondrogenesis in the ACH mouse model, with METTL16 overexpression promoting the proliferation of primary chondrocytes. METTL16 decreased ferroptosis both in vitro and in vivo and increased glutathione peroxidase 4 (GPX4) expression. METTL16 enhanced m6A modification of GPX4 mRNA and suppressed its degradation. Depletion of GPX4 abolished the effects of METTL16 on ACH mice and chondrocytes.
Overexpression of METTL16 improved bone growth and alleviated ferroptosis of chondrocytes by increasing m6A modification of GPX4 mRNA and thus GPX4 expression in chondrocytes. The METTL16/GPX4 axis may be a promising therapeutic approach for ACH treatment.
The dynamic evolution of signaling pathway remodeling and molecular regulation within the tumor microenvironment (TME) of hepatocellular carcinoma (HCC) play a critical role in the onset and progression of this malignancy. As chronic hepatitis progresses to cirrhosis and ultimately to HCC, the signaling pathways and TME show stage-specific characteristics that provide important insights into the therapeutic challenges and opportunities. In this review, we profiled the principal components of the HCC TME, along with pivotal signaling pathways, including the receptor tyrosine kinase (RTK) and extracellular signal-regulated kinase (ERK) pathways. Furthermore, we characterized the dynamic transformation of the TME from an inflammatory state in the hepatitis phase to a fibrotic state in the cirrhosis phase. Ultimately, we assessed the therapeutic potential of current HCC targets emphasizing emerging strategies for precision and personalized treatment.
Activation of autoreactive lymphocytes leads to cellular and tissue damage, which results in the development of autoimmune diseases. External environmental changes, such as chronic microbial infections, can alter the immune homeostasis and disrupt the balance of autoreactive T and B cells. In this review, we first summarize immune tolerance mechanisms of T and B cells, and then describe the breakthroughs of immune tolerance in T and B cells, followed by related autoimmune diseases. Furthermore, we explore how microbial infections can induce the production of autoreactive antibodies via carrier effects when the balance of autoreactive T and B cells is disrupted. These kinds of antibodies can lead to autoimmune diseases through molecular mimicry mechanisms. Our perspective provides a theoretical framework and novel insights into the mechanism of autoreactive antibodies in the pathogenesis of autoimmune diseases associated with microbial infections. This analysis may offer novel directions for drug discovery of autoimmune diseases.
Colorectal cancer (CRC) is a globally prevalent malignancy with rising incidence and mortality rates over the past decades. N6-methyladenosine (m6A) is the most abundant internal RNA modification in eukaryotes, and plays a pivotal role in post-transcriptional regulation. m6A is dynamically modulated by three core components, namely methyltransferases (writers), demethylases (erasers), and binding proteins (readers), which together govern the transcription, processing, translation, decay, and stability of mRNA. There has been accumulating evidence for the association of dysregulated m6A modification with CRC pathogenesis, metastasis, and therapeutic resistance. This review summarizes the biogenesis of m6A modification and its regulatory mechanisms, and discusses the dysregulation of m6A-related factors in CRC and the functional impacts. Most importantly, the review highlights the key roles of m6A modification in mediating CRC resistance to chemotherapy, targeted therapy, and immunotherapy. These insights may facilitate the development of novel therapeutic strategies for CRC.
In recent years, immunotherapy has gained increasing prominence in the treatment of hepatocellular carcinoma (HCC). However, effective immune-related biomarkers for HCC remain limited. In this study, both transcriptomic data and clinical information on HCC were obtained from The Cancer Genome Atlas (TCGA) database.
The TIMER and GEPIA databases were used to validate the association between WDR4 expression and immune infiltration. Additionally, clinical and pathological data from patients who underwent single-agent immunotherapy for HCC were collected from Hunan Provincial People’s Hospital (The First Affiliated Hospital of Hunan Normal University). The relationship between WDR4 expression levels, clinical pathological data, and patient prognosis was assessed using the Kruskal–Wallis test and Kaplan–Meier survival curve analysis. Spearman’s correlation analysis was utilized to confirm the relationship between WDR4, CD68, and PD-L1 in HCC tissue.
WDR4 was significantly upregulated in HCC tissues compared to para-carcinoma tissues (p < 0.001) and exhibited strong diagnostic potential. WDR4 expression showed significant associations with various immune cells, including macrophages (p < 0.001). Kaplan–Meier survival analysis revealed that patients with high WDR4 expression had shorter postoperative progression-free survival in the context of immunotherapy. Data from 37 patients who underwent postoperative single-agent immunotherapy for HCC demonstrated a significant correlation between WDR4 expression levels and disease-free survival (DFS), with strong statistical significance (log-rank p < 0.001).
WDR4 shows elevated expression in HCC tissues and is associated with immune infiltration, establishing it as a prognostic biomarker in HCC. Furthermore, the positive correlation observed between WDR4 and CD68, as well as PD-L1 (CD274), underscores its potential as a guiding factor in immunotherapeutic approaches for HCC.
The pathogenesis of multiple sclerosis (MS) is a complex topic that involves many individual aspects, several MS risk factors and various alterations in the immune system. In addition to the aspects still being researched in MS today, such as the activation of pro-inflammatory lymphocytes and macrophages, a particular scientific interest refers to the dysregulation of iron and to ferroptosis as an important form of programmed cell death (PCD), which is closely associated with this dysregulation. These two aspects are particularly significant because the new McDonald criteria include the diagnostic determination of so-called “paramagnetic rim lesions” (PRLs) in MS, which are areas of abnormal iron accumulation visible on magnetic resonance imaging (MRI) scans. Along with other diagnostic criteria, these PRLs are an important criterion for the diagnosis of MS. This review begins with an overall description of pathogenic risk factors and the dysregulation of the immune system. However, the priority of the review is the dysregulation of the cerebral iron balance and ferroptosis, as well as other forms of programmed cell death, which have also been demonstrated in MS: apoptosis, pyroptosis, necroptosis and NETosis. One of the main current findings on the forms of PCD in MS is their non-physiological occurrence in the brain’s own cells such as oligodendrocytes and neurons, and the extensive protection of pro-inflammatory cells from the occurrence of PCD. Emerging evidence indicates that ferroptosis occurs at an early stage in MS and appears to play a role in its pathogenesis. This review will present the distinct characteristics of the various PCDs in MS, alongside other pathological mechanisms, genetic factors, and current experimental approaches of therapeutic concepts that affect especially ferroptosis and pyroptosis. The aim is to provide an overview on iron dysregulation and forms of PCD as key aspects for understanding the pathogenesis of MS.
Hepatocellular carcinoma (HCC) is a leading cause of cancer-related mortality worldwide, owing to its molecular complexity and limited therapeutic options. Three-dimensional (3D) in vitro models more accurately recapitulate in vivo conditions than traditional two-dimensional (2D) models, due to their ability to accurately reproduce the tumor microenvironment (TME). Among these models, HepG2 cell-derived spheroids have become an important tool for drug screening, toxicity assessment, and liver cancer research. This review highlights the advantages and limitations of currently available 3D culture systems. In particular, special attention is given to the multifaceted role of leucine-rich repeat kinase 2 (LRRK2), a gene traditionally associated with neurological disorders but increasingly implicated in cancer, a kinase, which emerges as a promising therapeutic target in HCC since it regulates oxidative stress, lipid metabolism, and treatment responses, all of which contribute to tumor progression. Finally, we explore future directions, including organ-on-chip technologies and co-culture systems, which hold considerable promise for improving precision medicine and translational research in HCC.
Liver fibrosis, the end-stage pathological state of many liver diseases, is primarily driven by the activation of hepatic stellate cells (HSCs) and collagen deposition resulting from various pathogenic causes. Thrombospondin-2 (THBS2), a secreted extracellular matrix glycoprotein encoded by the TSP gene family, has been found to activate the TLR4-transforming growth factor-β (TGF-β)/FAK signaling axis and HSCs through autocrine signalling, thereby contributing to the development of liver fibrosis. Latexin (LXN), the only known zinc-dependent metallocarboxypeptidase inhibitor in humans, has not yet been studied for its role in liver fibrosis is yet to be studied.
In this study, we used adeno-associated virus 9 (AAV9) to generate a mouse model of liver fibrosis with LXN knockdown and used siLXN to knock down the LXN gene in the human hepatic stellate cell line LX-2. The mechanisms underlying the association between LXN and hepatic fibrosis progression were investigated using quantitative polymerase chain reaction, western blot, immunohistochemistry, and immunofluorescence staining.
LXN knockdown reduced carbon tetrachloride (CCl4)-induced liver injury and suppressed activation of hepatic stellate cells, while also inhibiting the expression of α-SMA and collagen I. Furthermore, LXN demonstrates a substantial positive correlation with THBS2, and LXN knockdown was capable of downregulating THBS2.
The LXN-THBS2 signaling axis may promote liver fibrosis progression by inducing the activation of HSCs.
High-grade ovarian cancer (HGOC) is a heterogeneous and aggressive malignancy with a tumor microenvironment (TME) that suppresses immune responses, limiting immunotherapy efficacy. Ferroptosis, an iron-dependent form of regulated cell death, has emerged as a potential therapeutic target.
We investigated the immunomodulatory effects of the ferroptosis inducer RAS-Selective Lethal 3 (RSL3) in four HGOC cell lines (ES-2, OVCAR-5, HEY, PEO-1) using flow cytometry and lactate dehydrogenase (LDH) release assays.
RSL3 modulated Natural Killer (NK) ligand expression in a cell line-dependent manner, resulting in differential susceptibility to NK cell-mediated cytotoxicity. OVCAR-5 cells became more susceptible to NK cell killing after treatment, whereas HEY cells showed reduced susceptibility, and ES-2 and PEO-1 cells exhibited minimal changes.
Ferroptosis induction alone does not consistently enhance NK cell-mediated cytotoxicity in HGOC cells. These findings underscore the heterogeneity of tumor responses and highlight the need for further studies, particularly in in vivo models, to elucidate mechanisms linking ferroptosis to immune recognition and thereby inform therapeutic development.
Altered homeostasis of the ocular surface microenvironment is a hallmark of dry eye disease (DED). The alpha-7 nicotinic acetylcholine receptor (α7nAChR) plays a key role in DED pathophysiology. In this study, we established a rabbit model of DED using scopolamine hydrobromide (Scop) to determine the effect of electroacupuncture (EA) on ocular surface damage in DED and to explore its underlying mechanisms.
New Zealand White rabbits (1.5–2.0 kg) were subcutaneously administered Scop for 21 days prior to EA treatment. After 35 days, the homeostasis of the ocular surface microenvironment was evaluated using the Schirmer I test (SIT), tear break-up time (BUT), corneal fluorescein (FL) staining, and measurement of tear osmolarity. The expression levels of ACh, α7nAChR, and high mobility group box 1 (HMGB1) were detected via histopathological examination of the cornea, lacrimal glands, and conjunctiva, combined with immunohistochemistry and western blotting. Additionally, protein chip technology was used to determine the expression levels of downstream factors.
EA stimulation significantly improved the homeostasis of the ocular surface microenvironment, as evidenced by increased SIT values and BUT, reduced corneal FL intensity, and decreased tear osmolarity. It also alleviated pathological damage to the cornea, conjunctiva, and lacrimal glands; upregulated the expression of ACh and α7nAChR; and downregulated the expression of HMGB1 and related inflammatory factors. However, these changes were reversed following administration of α-Bungarotoxin.
EA stimulation improves ocular surface homeostasis and reduces inflammation in DED, potentially via activation of the α7nAChR signaling pathway, which in turn inhibits the expression of HMGB1 and inflammatory factors.
Therapeutic strategies for ocular diseases are undergoing a transformative shift from symptom management to regenerative and disease-modifying approaches. This review highlights the development of neurotrophin receptor agonists—including recombinant nerve growth factor (NGF) (cenegermin), peptidomimetics (e.g., REC-0559, tavilermide), and synthetic microneurotrophins (BNN27, ENT-A010)—that target tropomyosin receptor kinases (TrkA/TrkB) and the p75 neurotrophin receptor (p75NTR) pathways to promote neuronal survival, synaptic plasticity, and tissue repair in neurotrophic keratitis, dry eye disease, and retinal degenerations. Parallel advances in peptide-based therapies address vascular and inflammatory pathologies: UPARANT and its derivatives modulate urokinase plasminogen activator receptor (uPAR)/formyl peptide receptor (FPR) signaling to inhibit angiogenesis and inflammation in diabetic retinopathy, whereas sphingosine 1 phosphate (S1P)-S1PR3 pepducins and integrin antagonists (risuteganib, THR-687, OTT166) offer multi-targeted strategies to stabilize the blood-retinal barrier and mitigate neovascularization. Innovations in drug delivery, such as dendrimer-peptide conjugates, enhance the stability and bioavailability of these agents. Further, senolytic therapies (e.g., UBX1325, procyanidin C1) are emerging as a promising approach for age-related and diabetic retinal diseases by clearing senescent cells and attenuating senescence-associated secretory phenotype (SASP)-driven inflammation. Together, these approaches exemplify a paradigm of “mimicking nature to modulate vision”, leveraging molecular insights to develop therapies that restore rather than merely preserve ocular function. While clinical validation is ongoing, the convergence of neurotrophic support, vascular modulation, and senescence targeting heralds a new era in precision ophthalmology.
This review examines the applications of natural coumarins in the context of bone and joint disorders. It provides comprehensive coverage of their fundamental pharmacology, the epidemiology and pathological mechanisms of osteoarticular diseases, as well as the pharmacological actions, clinical applications, future prospects, and ongoing challenges associated with these compounds. Through systematic analysis of relevant studies, we summarize the basic characteristics of natural coumarins, including their chemical structures, metabolic pathways, and pharmacological activities. We also examine their mechanisms of action in conditions such as osteoarthritis, rheumatoid arthritis, and osteoporosis, with a particular emphasis on their anti-inflammatory and cartilage-repairing properties. Furthermore, we summarize current clinical trials, formulation development, delivery strategies, and safety evaluations. We also explore potential new indications, combination therapies, and personalized treatment approaches. Several controversies and ongoing challenges are discussed, including variability in dose-dependent efficacy, divergent therapeutic outcomes across diseases, and concerns regarding long-term safety. This review aims to provide a foundational reference for further research and clinical application of natural coumarins in the treatment of bone and joint disorders.
Accurate biodosimetry is essential for effective radiological triage, precise clinical monitoring, and assessment of risks from diagnostic exposures. Immunofluorescent detection of phosphorylated histone H2AX (γH2AX) and p53-binding protein 1 (53BP1) DNA double-strand break repair foci provides high sensitivity for radiation dose assessment within the first hours after exposure. However, inter-laboratory reproducibility of γH2AX/53BP1-based biodosimetry remains limited, and the contribution of pharmacological modifiers is unresolved.
Here, we conducted an inter-laboratory comparison of in vitro radiation dose–response relationships of foci yields measured in cryopreserved umbilical cord blood lymphocytes (UCBLs) and freshly isolated peripheral blood lymphocytes (PBLs) from healthy donors using fluorescent microscopy with emphasis on workflow harmonization and reproducibility across laboratories.
Under low-dose γ irradiation, both UCBLs and PBLs exhibited a strong linear, dose-dependent induction of γH2AX, 53BP1, and co-localized foci, with co-localization emerging as the most sensitive endpoint. γH2AX pan-nuclear staining was observed exclusively in UCBLs and functioned as a distinct endpoint under the examined conditions. Under harmonized low-dose conditions, calyculin A at a non-toxic concentration of 1 nM did not provide measurable stabilization or enhancement of ionizing radiation-induced foci (IRIF) yields. Although IRIF yields differed between the two laboratories, dose–response slopes were highly concordant, demonstrating reproducibility under harmonized experimental conditions.
These findings demonstrate that inter-laboratory reproducibility of γH2AX/53BP1-based biodosimetry is achieved primarily through disciplined workflow harmonization rather than through pharmacological enhancement. By reinforcing assay reproducibility and biological consistency, this work supports the translational applicability of IRIF-based biodosimetry for broader application in radiation exposure assessment.
Rotator cuff injuries are common musculoskeletal disorders and are frequently complicated by impaired tendon–bone healing and high re-tear rates after surgical repair. Exosomes derived from adipose-derived stem cells (ADSCs) have shown regenerative potential through paracrine mechanisms; however, the role of exosomal insulin-like growth factor 1 (IGF1) in tendon–bone healing remains unclear.
Exosomes were isolated from rat ADSCs with or without lentiviral knockdown of IGF1. A rat supraspinatus tendon tear and repair model was established, and 200 μg of exosomes was administered systemically post-surgery. Tendon–bone healing was evaluated at 8 weeks post-operation using histological, immunohistochemical, and micro-computed tomography analyses. Early molecular responses were assessed at 1-week post-surgery by Western blot and RT-qPCR. Angiogenic markers (vascular endothelial growth factor (VEGF), CD31, α-SMA), inflammatory cytokines (interleukin (IL)-1β, IL-18), pyroptosis-related proteins (gasdermin D N-terminal fragment (GSDMD-N)), and NLRP3 inflammasome components were examined.
ADSC-derived exosomes significantly enhanced bone mineral density, fibrocartilage formation, vascularization, and biomechanical strength at the tendon–bone interface. These effects were accompanied by reduced inflammatory cytokine expression, inhibition of pyroptosis, and suppression of NLRP3 inflammasome activation. In contrast, exosomes derived from IGF1-deficient ADSCs exhibited markedly reduced therapeutic efficacy, with attenuated angiogenic, anti-inflammatory, and anti-pyroptotic effects.
Exosomal IGF1 plays a critical role in promoting angiogenesis, suppressing inflammation and pyroptosis, and improving structural and biomechanical outcomes during tendon–bone healing. IGF1-enriched ADSC-derived exosomes represent a promising therapeutic strategy for enhancing rotator cuff repair.
In recent years, drug-resistant influenza viruses have emerged frequently, making influenza a persistent and serious public health burden. Therefore, potential anti-influenza virus drugs are urgently needed. Nanobodies, variable domains of heavy-chain antibodies (VHHs), have the advantages of easy preparation, excellent solubility, deep tissue penetration, and weak immunogenicity; thus, they have broad application prospects in the fields of basic research and drug development. However, its short half-life and low stability limit its clinical therapeutic application. Fenobody is an engineered display platform with the ability to present multimerized nanobodies on the surface of ferritin to overcome these disadvantages and increase its potency.
In this study, we engineered a fenobody displaying multimerized VHH against haemagglutinin (HA) of influenza virus (A/California/07/2009(H1N1), pdm09) on the surface of ferritin by using the property of SpyTag to spontaneously bind to SpyCatcher, named ferritin-NP-VHH.
Compared with VHH alone, ferritin-NP-VHH improved the cross-neutralizing activity, stability and affinity for influenza virus in vitro and prolonged its half-life in vivo.
These results suggest that the implementation of genetic engineering technology to construct multimerized anti-influenza virus nanoparticles provides new tools to control infection with influenza virus.
Chronic aerobic exercise is known to modulate oxidative stress, yet comparative analyses across multiple tissues remain limited. This study aimed to evaluate the effects of different training durations on oxidative damage and membrane fluidity in metabolically active tissues, including skeletal muscle, heart, and brain.
Forty male Sprague–Dawley rats were randomly assigned to four groups (n = 10/group): control (CON), and aerobic treadmill training for 1 week (1W), 4 weeks (4W), and 12 weeks (12W). Training consisted of four 60-minute sessions per week, alternating intensities at 35% and 80% of maximal velocity. Quadriceps skeletal muscle, heart, and brain were collected. Oxidative damage was assessed via malondialdehyde and 4-hydroxyalkenals (MDA + 4-HDA) and protein carbonyl content. Membrane fluidity was evaluated in plasma and mitochondrial membranes using fluorescence spectroscopy. Statistical analyses were performed using one-way analysis of variance followed by Tukey post-hoc tests.
Aerobic training significantly reduced MDA + 4-HDA levels in skeletal muscle and heart compared with control values, with a downward trend in brain tissue that did not reach statistical significance. Protein carbonyls decreased significantly in skeletal muscle at 4W and 12W, but remained unchanged in heart and brain (despite lower mean values observed in all training groups compared with the control). Plasma membrane fluidity declined significantly in all tissues, especially at 4W and 12W, indicating structural remodeling. Mitochondrial membrane fluidity values were lower in heart and skeletal muscle across the different training groups, with statistically significant differences observed only and remarkably in skeletal muscle compared with control values; meanwhile, values remained stable in brain tissue. These findings reveal tissue-specific biochemical and biophysical adaptations to aerobic training.
Chronic aerobic treadmill training induces protective adaptations against oxidative stress in skeletal muscle, heart, and brain. The reduction in lipid peroxidation and protein oxidation, along with changes in membrane fluidity, reflects enhanced cellular resilience and structural integrity. These results support the role of sustained aerobic exercise as a non-pharmacological strategy to mitigate oxidative damage and promote tissue health. The rat model used provides translational relevance for understanding exercise-induced protection mechanisms.
The metabolic profile of cancer includes changes in energy metabolism and biosynthetic (plastic) metabolism, and redox balance of tumor cells. This study aimed to identify clinically significant salivary metabolic features associated with breast cancer phenotypes.
This study included 660 patients with breast cancer (age 54.6 ± 12.7 years) and 127 healthy volunteers (49.3 ± 14.2 years). Saliva samples were collected from all participants, strictly before the initiation of treatment, and the biochemical composition of saliva was determined, including indicators of antioxidant system activity, lipid profile, cytokines, and free amino acids.
Salivary metabolic features correlated with the breast cancer phenotype. In particular, for luminal A breast cancer, which has the most favorable prognosis, the presence of an active inflammatory process in saliva (C-reactive protein +136.6%, p < 0.0001; IL-1β +317.7%, p = 0.0004) and a pronounced immune anti-inflammatory response (INF-γ +79.1%, p = 0.0004) were shown. In contrast, for triple-negative breast cancer, a low anti-inflammatory response (INF-γ –4.1%) and active cell proliferation (glutamine +45.0%, p = 0.0342) were shown, which correlated with the disease severity, low immunogenicity, and the least favorable prognosis for this subtype of breast cancer.
Overall, salivary composition reflects systemic metabolic changes in breast cancer, which makes it possible to construct a metabolic portrait of breast cancer across distinct phenotypes.
Acute myeloid leukemia (AML) is a hematological malignancy of the myeloid lineage with poor clinical outcomes due to limited targeted therapies. This study elucidates the role of inositol 1,4,5-trisphosphate receptor type 2 (ITPR2) in regulating cell apoptosis by modulating mitochondrial calcium (Ca2+) levels and underscores the clinical significance of ITPR2.
ITPR2 expression in patients with AML compared with that of healthy controls using the quantitative real-time PCR (RT-qPCR) method. The role of ITPR2 in AML and its association with immune infiltration levels were investigated through bioinformatics analyses. ITPR2 knockdown in Tohoku Hospital Pediatrics-1 (THP-1) cells were achieved using small interfering RNA (siRNA) and 2-aminoethyl diphenylborinate (2-APB), followed by comprehensive molecular characterization employing RT-qPCR, western blotting (WB), cell counting kit-8 (CCK-8) assays, and flow cytometry.
ITPR2 was validated as being highly expressed in patients with AML, and this expression correlated with risk stratification and poor prognosis. Functional enrichment analysis revealed that ITPR2 is involved in Ca2+ signaling pathways and mitochondrial-related biological processes, and its expression level was negatively correlated with immune infiltration levels. Knockdown of ITPR2 or inhibition of its activity with 2-APB reduced Ca2+ ion concentrations in both the cytoplasm and mitochondria, leading to mitochondrial dysfunction (characterized by elevated intracellular reactive oxygen species (ROS) levels, reduced mitochondrial membrane potential (MMP) levels, and reduced mitochondrial DNA copy number) and eventually AML cell apoptosis.
ITPR2 facilitates AML progression via the Ca2+-mitochondrial axis and may serve as a prognostic factor and potential therapeutic target.
Recent studies have identified impaired renal gluconeogenesis as a hallmark of chronic kidney disease. Triptolide is a natural compound widely used in China for the treatment of renal diseases. This study investigated whether triptolide mitigates renal fibrosis by promoting renal gluconeogenesis.
Renal fibrosis was induced in vivo by unilateral ureteral obstruction (UUO) surgery in mice. Transforming growth factor-β (TGF-β)-stimulated human kidney-2 (HK-2) cells were used as an in vitro model to investigate renal fibrosis. Metabolomics, western blotting, immunohistochemistry (IHC), and metabolic assays were performed to investigate the underlying mechanisms.
Triptolide reduced the expression of several fibrotic markers in the kidneys of UUO mice. Metabolomic analysis revealed enhanced renal gluconeogenesis following treatment with triptolide, which was confirmed by analyzing gluconeogenic enzyme expression and lactate concentration in UUO kidneys. The pro-gluconeogenic effect of triptolide was further confirmed in TGF-β-stimulated HK2 cells. Inhibition of phosphoenolpyruvate carboxykinase 1 (PCK1) reversed the anti-fibrotic and pro-gluconeogenic effects of triptolide in TGF-β-stimulated HK2 cells. We further demonstrated that peroxisome proliferator-activated receptor-gamma co-activator 1 alpha (PGC1α) expression was downregulated in TGF-β-stimulated HK2 cells and UUO kidneys, and that triptolide reversed this downregulation. Moreover, the PGC1α inhibitor reversed the effect of triptolide on PCK1 expression and glucose metabolism. Finally, IHC analysis revealed that triptolide inhibited histone lactylation in UUO kidneys, which was associated with a decreased production of inflammatory factors and reduced macrophage infiltration.
Triptolide may inhibit renal fibrosis by increasing the PGC1α/PCK1 axis, thereby promoting renal gluconeogenesis. This cascade may reduce histone lactylation and renal inflammation, providing a mechanistic pathway for its anti-fibrotic effect.
Escherichia coli is the leading cause of urinary tract infections (UTIs), and the increasing prevalence of antimicrobial resistance represents a major public health concern. The dissemination of multidrug-resistant uropathogenic E. coli (UPEC), frequently harboring transferable resistance determinants, poses an urgent clinical challenge.
This study investigated the prevalence of β-lactamase genes (blaTEM, blaSHV, blaCTX-M, blaCMY, and blaDHA) and plasmid-mediated quinolone resistance genes (qnrA, qnrB, qnrC, qnrD, and qnrS) in 86 imipenem-non-susceptible UPEC isolates using multiplex and single PCR assays. Gene distribution and co-occurrence were examined across E. coli phylogenetic groups, and pairwise associations were evaluated using correlation analysis. Principal component analysis (PCA) was applied to explore global relationships between antibiotic susceptibility profiles, extended-spectrum β-lactamase (ESBL) phenotype, and resistance determinants.
Overall, 74.4% of imipenem-non-susceptible isolates carried at least one β-lactamase gene. blaTEM was the most prevalent (62.8%), followed by blaCMY II (12.8%). blaSHV, blaCTX-M group I, and blaCTX-M group II showed comparable prevalence (10.5% each). The B2 phylogroup showed the greatest diversity of β-lactamase profiles, with phylogroup E representing the second most frequent reservoir. Among quinolone resistance genes, qnrB was the most prevalent (20.9%), followed by qnrD (5.8%), qnrS (4.7%), qnrA (3.5%), and qnrC (1.2%). All qnrC-positive isolates were resistant to all tested quinolones. No statistically significant associations were observed between β-lactamase genes and qnr genes. Significant within-class correlations were detected for blaCTX-M group II–blaCMY II (φ = 0.893, q = 9.33 × 10–9) and qnrC–qnrA (φ = 0.57, q = 0.374).
A high prevalence of β-lactamase and qnr determinants was observed among imipenem-non-susceptible UPEC, primarily driven by blaTEM and qnrB, frequently detected in B2 isolates. The co-occurrence and correlation of multiple resistance genes highlight the complexity of resistance architectures and underscore the need for ongoing molecular surveillance and strengthened antimicrobial stewardship to limit the dissemination of resistant UPEC.
Purple sweet potato anthocyanins (PSPAs), a class of dietary flavonoids, have shown anticancer potential. However, their ability to induce ferroptosis in T-cell acute lymphoblastic leukemia (T-ALL) remains unexplored. This study aimed to investigate whether PSPAs can trigger ferroptosis in T-ALL cells and to elucidate the underlying mechanisms.
Jurkat T-ALL cells were treated with PSPAs, and cell viability, reactive oxygen species (ROS), lipid peroxidation, intracellular Fe2+, and expression of ferroptosis-related proteins (glutathione peroxidase 4 (GPX4), solute carrier family 7 member 11 (SLC7A11), nuclear factor erythroid 2-related factor 2 (Nrf2)) were assessed. Ferrostatin-1 was used to verify ferroptosis involvement. Ultrastructural changes were examined by electron microscopy. Molecular docking was performed to evaluate PSPA binding to SLC7A11, and in vivo efficacy was tested in T-ALL xenograft mice.
PSPAs exhibited significant cytotoxicity in Jurkat cells, which was reversed by ferrostatin-1, indicating ferroptosis involvement. Treatment elevated ROS and lipid peroxidation, increased intracellular Fe2+, and downregulated GPX4 and SLC7A11 without altering Nrf2, suggesting that SLC7A11 may be directly targeted. Electron microscopy revealed hallmark ferroptotic changes, including increased mitochondrial membrane density, loss of cristae, and rupture of the outer membrane. Molecular docking demonstrated strong binding of four PSPA components to multiple residues of SLC7A11, including Cys158, a key functional site. In vivo, PSPAs markedly inhibited tumor growth in T-ALL xenograft mice, achieving up to 75% suppression, as evidenced by histological analysis showing disrupted tumor architecture and cell membrane rupture.
This study provides the first evidence that PSPAs induce ferroptosis in T-ALL through modulation of the SLC7A11/GPX4 pathway. These findings reveal new mechanistic insights into ferroptosis in T-ALL and highlight PSPAs as safe, naturally derived therapeutic agents with promising therapeutic potential for leukemia.
Bronchopulmonary dysplasia (BPD) is a chronic lung disease in premature infants. Neonatal hyperoxia induces a BPD-like phenotype and lung cell senescence in rodents. In our 3-day hyperoxia model, senescent cells were predominantly lung macrophages, with their abundance peaking at postnatal day 7 (pnd7). However, the molecular and functional characteristics of these senescent macrophages remain undefined.
We reanalyzed a scRNA-seq dataset (GSE207866) generated from senescent lung cells isolated at pnd7 (SD7) following neonatal hyperoxia. Hierarchical clustering combined with manual annotation was used to compare transcriptional profiles with age-matched air-exposed controls (AirD7) and hyperoxia-exposed mice without senescent-cell enrichment (O2D7). Key molecular findings were validated by immunofluorescence. In vivo, neonatal mice received daily injections of the pyruvate dehydrogenase kinase inhibitor, dichloroacetate (DCA) from pnd4 to pnd6, and a senolytic cocktail consisting of quercetin and dasatinib from pnd4 to pnd14, following 3 days of hyperoxia exposure.
Macrophages accounted for 65.90% of senescent cells in the SD7 group. Seven macrophage clusters were identified, enriched in M1-like and alveolar macrophage phenotypes. Two major clusters (clusters 0 and 1), together representing nearly half of all senescent macrophages, exhibited strong expression of genes associated with innate immunity, inflammation, and DNA damage responses. These clusters also showed a shift toward glycolysis, the pentose phosphate pathway, and glutamine metabolism, with reduced reliance on β-oxidation. Administration of DCA activated pyruvate dehydrogenase and attenuated hyperoxia-induced macrophage senescence and lung injury. Pathway enrichment analyses revealed enhanced metal-handling pathways, immune and stress signaling (including p38 mitogen-activated kinase, ataxia-telangiectasia mutated, and mechanistic target of rapamycin), apoptosis, and RNA regulatory processes. Conversely, genes involved in reactive oxygen species detoxification, DNA repair, phagocytosis, cytoskeletal organization, and cell adhesion were downregulated. Notably, reducing senescent cells by a senolytic cocktail during the alveolar stage mitigated hyperoxia-induced persistent lung injury.
Neonatal hyperoxia drives the emergence of a heterogeneous population of senescent macrophages characterized by metabolic reprogramming and dysregulated signaling pathways, which contribute to the development and persistence of lung injury.
Malignant gliomas remain largely refractory to current therapies, in part because abnormal tumor vasculature and a disrupted blood-brain barrier limit intratumoral drug delivery and cause severe tumor-associated edema. The oncolytic adenovirus KD01 is a conditionally replicating adenovirus type 5 with a 27-bp deletion in Early Region 1A (E1A) and a truncated BH3-Interacting Domain Death Agonist (tBID) expression cassette inserted into the E3 region. This study investigated whether bevacizumab-induced vascular normalization would enhance the delivery and efficacy of KD01 in gliomas.
The oncolytic activity and mitochondrial effects of KD01 were evaluated in human glioma cell lines using cell viability assays, JC-1 staining, quantitative real-time-polymerase chain reaction (qRT-PCR), and western blotting for BID/tBID. An orthotopic LN229 nude mouse model was used to assess a sequential bevacizumab→KD01 regimen. Mice were randomized to receive PBS, bevacizumab, KD01, or combination treatment. Body weight and survival were recorded. Tumor cell proliferation (Ki-67), tumor vasculature (CD31), brain water content (ΔWater%), serum biochemistry, coagulation parameters, and organ weights were analyzed to evaluate antitumor activity, edema, and systemic safety.
KD01 induced robust dose- and time-dependent cytotoxicity in glioma cells and caused marked mitochondrial depolarization, accompanied by increased BID mRNA expression, loss of full-length BID, and accumulation of tBID. In the orthotopic LN229 model, bevacizumab administered 48 h before KD01 significantly improved overall outcomes compared with either monotherapy. The bevacizumab→KD01 group showed improved preservation of body weight, pronounced prolongation of survival, and the lowest Ki-67 labeling index. This group also exhibited reduced brain water content (ΔWater%), consistent with sparser CD31-positive vessels resulting from vascular normalization and oncolysis, indicating effective attenuation of tumor-associated edema. Serum liver and kidney function tests, platelet counts, coagulation indices, and major organ weights were comparable across treatment groups, suggesting no additional systemic toxicity associated with combination treatment.
KD01 exerts potent tBID-mediated mitochondrial oncolytic activity against glioma cells. When used as a priming strategy, transient vascular normalization induced by bevacizumab enhanced the intratumoral efficacy of KD01 in an orthotopic glioma model while maintaining a favorable safety profile. These findings support a simple, sequence-dependent combination approach integrating anti-VEGF therapy with oncolytic virotherapy for the treatment of malignant gliomas.
Arsenic trioxide (ATO) is a cornerstone of acute promyelocytic leukemia (APL) therapy but induces severe gut microbiota dysbiosis, limiting its efficacy and safety. This study investigated whether adjunctive Bifidobacterium pseudolongum (BP) could mitigate these adverse effects and enhance therapeutic outcomes.
16S rRNA gene sequencing data of gut microbiota were obtained from a cohort of 22 APL patients treated with ATO-based regimens (20 of 22 data were obtained and analysis further), accessible under BioProject ID PRJNA935705. To evaluate the within-sample microbial community richness and evenness, alpha and beta diversity indices were calculated. Using a murine APL model, we compared ATO monotherapy with ATO+BP co-treatment. Analyses included fecal metagenomic sequencing, single-cell RNA sequencing (sc-RNA-seq), flow cytometric immune profiling, and assessment of intestinal tight junction proteins (claudin-1, occludin, and ZO-1) via immunofluorescence.
ATO treatment significantly reduced gut microbial diversity and depleted beneficial taxa. Sc-RNA-seq data showed that ATO could orchestrate the APL immune microenvironment mainly through functional activation of CD8+ T cells and monocytes. BP supplementation restored microbial homeostasis and synergistically enhanced ATO’s antileukemic effect, reducing the leukemic burden in peripheral blood by 72% and in bone marrow by 64% compared to ATO alone. Mechanistically, BP preserved intestinal barrier integrity by upregulating tight junction protein expression and modulated anti-tumor immunity, notably increasing bone marrow CD8+ T cells by 2.21-fold.
BP is an effective adjunct to ATO therapy, counteracting gut dysbiosis, intestinal damage, and the immune microenvironment while synergistically improving antileukemic efficacy. Targeting the gut–leukemia axis with BP represents a promising strategy for improving the precision and safety of APL treatment.
Metabolic reprogramming is a hallmark of the pathogenesis and progression of endometrial carcinoma (EC). This study comprehensively analyzed the expression profiles of glycine, serine, and threonine (Gly/Ser/Thr) metabolism–related genes in EC. We also established a robust prognostic model and developed a molecular subtyping framework that integrates metabolic and immune characteristics based on the identified prognostic genes. The aims of this work are to enhance diagnostic precision and improve clinical management strategies for patients with EC.
Untargeted metabolomic analysis was performed on 35 EC and 15 normal tissues. The Cancer Genome Atlas (TCGA) transcriptomic data were integrated with weighted gene co-expression network analysis (WGCNA) to identify EC-related metabolic genes and construct a prognostic model using Cox proportional hazards and least absolute shrinkage and selection operator (LASSO) regression analyses. The model was validated using an independent proteomic and single-cell dataset from our institution. Consensus clustering classified patients into three molecular subtypes, which were further characterized by gene set variation analysis (GSVA) and profiling of immune infiltration. Finally, key prognostic genes were validated by reverse transcription quantitative polymerase chain reaction (RT-qPCR) in EC and normal endometrial epithelial cells.
Metabolomic analysis revealed significant enrichment of the Gly/Ser/Thr metabolic pathways. WGCNA identified a tumor-associated metabolic module among 1741 pathway-related genes. A prognostic model comprising methylenetetrahydrofolate dehydrogenase 2 (MTHFD2), ribosomal protein S6 kinase A1 (RPS6KA1), and cyclin-dependent kinase inhibitor 2A (CDKN2A) was subsequently established. Consensus clustering based on risk scores stratified EC patients into three molecular subtypes: immunometabolic-suppressed (C1), proliferative-immunobalanced (C2), and immune-activated (C3). The C1 subtype had the poorest prognosis and was characterized by metabolic suppression and immune evasion. The C2 subtype showed a favorable prognosis and was defined by a “proliferation–immune balance” in which high proliferative activity coexisted with strong anti-tumor immunity. The C3 subtype was also associated with a favorable outcome, driven by upregulated DNA repair and oxidative phosphorylation pathways alongside infiltration of immune-active cells. RT-qPCR confirmed significant differences in the mRNA expression of MTHFD2, RPS6KA1, and CDKN2A between normal and EC cells (p < 0.05).
This study developed a Gly/Ser/Thr pathway–based prognostic model for EC, based on the expression of MTHFD2, RPS6KA1, and CDKN2A as novel biomarkers. The resulting patient stratification framework holds significant clinical potential for guiding precise and personalized management of EC.
Long-QT syndrome type 2 (LQTS2), which is associated with life-threatening cardiac arrhythmias, is caused by pathogenic heterozygous loss-of-function mutations in the KCNH2 gene. This gene encodes the pore-forming Kv11.1 α-subunit of the ion channel that carries the rapid delayed rectifier potassium current (IKr). Pathogenic loss-of-function mutations reduce the amplitude of IKr, thereby prolonging the action potential (AP) of ventricular cardiomyocytes, and in turn, the QT interval of the electrocardiogram (ECG). The aim of the present in silico study was to test the extent to which allele-specific suppression (‘silencing’) of the mutant KCNH2 allele can alleviate the effects of dominant-negative LQTS2 mutations.
Two recent and comprehensive models of the electrical activity of a single human ventricular cardiomyocyte, i.e., the ‘Bartolucci–Passini–Severi model as published in 2020’ and the ‘Tomek–Rodriguez model following the O’Hara–Rudy dynamic (ORd) model’ (known as the BPS2020 and ToR–ORd models, respectively) were used to assess the effects of mild and severe LQTS2 mutations on the AP duration at 90% repolarization (APD90) and the APD90 restitution obtained with an S1-S2 pacing protocol.
For severe mutations, the mutation-induced prolongation of the APD90 at a stimulation rate of 1 Hz is reduced from 166% to 99% in the BPS2020 model and from 111% to 71% in the ToR–ORd model upon 70% suppression of the mutant allele. For mild mutations, this prolongation is reduced from 77% to 44% and from 57% to 34%, respectively. An even greater effect is observed when the mutant KCNH2 allele is inhibited by up to 90%, but the greater suppression is only marginal for mild mutations. The steepness of the mutant APD90 restitution curves is considerably reduced upon suppression, which may exert an anti-arrhythmic effect.
Silencing of the mutant allele can substantially, but only partially, counteract the effects of mild or severe LQTS2 mutations on IKr. Allele-specific inhibition of the mutant KCNH2 allele alone is not sufficient to fully treat the effects of LQTS2 mutations and should be accompanied by a replacement gene therapy, creating a suppression-and-replacement (“SupRep”) gene therapy.
Aging is frequently accompanied by chronic, low-grade inflammation, often referred to as “inflammaging”, which contributes to functional decline of multiple organs including the liver. The NLRP3 inflammasome has emerged as a key mediator of age-related inflammation; however, its pharmacological inhibition in the context of hepatic aging remains insufficiently explored. In this study, we investigated the effects of the selective NLRP3 inflammasome inhibitor MCC950 on inflammatory responses in the liver of aged mice.
Aged C57BL/6 mice (18 months old) were administered MCC950 intraperitoneally for four weeks, and liver tissues were analyzed for inflammatory and stress-related markers.
MCC950 treatment significantly reduced hepatic expression of NLRP3, caspase-1 activation, and IL-1β production, accompanied by a decrease in proinflammatory cytokines such as p-STAT3. Histological analysis demonstrated attenuation of age-associated hepatic inflammatory infiltration and improved tissue architecture. Furthermore, MCC950 administration restored autophagy-related proteins (LC3B, p62) indicating broader protective effects on liver homeostasis.
These findings suggest that NLRP3 inflammasome inhibition with MCC950 alleviates age-associated hepatic inflammation and may represent a potential therapeutic strategy for mitigating inflammaging and preserving liver function in the elderly.