Our previous study demonstrated a potent synergistic antitumor effect between 1 nM digoxin and 1 μM cisplatin in cervical cancer cells, which was dependent on the modulation of the signaling pathway involving Na, K-ATPase/Src. Because of the relationship between Na, K-ATPase/Src and the modulation of oxidative stress, the aim of this study was to demonstrate the effect of combined treatment with digoxin and cisplatin on oxidative stress parameters in HeLa cells and the effect of treatment in non-tumor cell lines. Analysis of oxidative stress parameters revealed a significant reduction in cellular antioxidant defense mechanisms, SOD activity, and content of reduced total thiol groups. Additionally, there was an increase in H2O2 content within 24 h, confirming the elevated cellular oxidative stress during the combined treatment. Furthermore, lipid peroxidation and lipid droplet formation levels were evaluated, showing an increase in these parameters after 48 h of treatment, consistent with oxidative stress. Compared to the other tested cell lines, the combined treatment did not significantly alter cell viability compared to the control. Our data demonstrate that the synergistic antitumor effects of digoxin and cisplatin in HeLa cells are tightly associated with increased oxidative stress via activation of the Na, K-ATPase/Src/ROS signaling pathway, may resulting in antiproliferative and pro-apoptotic effects. These findings indicate that oxidative stress is a key component of the cellular response underlying the synergistic antitumor effects of digoxin and cisplatin.

Ferroptosis is a regulated form of cell death in which disturbed iron metabolism, accumulation of peroxidized membrane lipids, and insufficient glutathione peroxidase 4 (GPX4) activity converge to injure vascular cells. In atherosclerosis, its effects are highly dependent on cellular context rather than being uniform across the vessel wall. In macrophages, ferroptotic injury strengthens inflammatory signaling and contributes to necrotic-core growth; in vascular smooth muscle cells, it impairs mitochondrial fitness, contractile phenotype maintenance, and extracellular matrix support, thereby weakening the fibrous cap; and in endothelial cells, it disrupts redox homeostasis and barrier function, facilitating lipid entry and early lesion formation. This review synthesizes recent advances in the molecular regulation of ferroptosis in atherosclerosis and emphasizes how its functional consequences differ among plaque-resident cell types. We also discuss natural products and traditional Chinese medicine-derived compounds as promising modulators of ferroptosis-related pathways, owing to their multi-target actions and potential suitability for chronic vascular intervention. By connecting mechanistic evidence with therapeutic implications, we propose that cell-type-aware regulation of ferroptosis, rather than indiscriminate suppression, may offer a rational strategy for plaque stabilization and cardiovascular risk reduction. Significance: Ferroptosis is increasingly recognized as an important contributor to atherosclerosis, but its vascular effects vary substantially among endothelial cells, macrophages, and vascular smooth muscle cells. This review analyzes how cell-specific differences in iron handling, lipid peroxidation, and antioxidant defenses shape plaque initiation, inflammatory progression, and fibrous-cap stability. We further summarize evidence that natural compounds can modulate these ferroptosis-related networks and restore redox and cellular homeostasis. By linking vascular cell heterogeneity with natural product-based intervention strategies, this review provides a mechanistic and translational framework for more precise targeting of ferroptosis in atherosclerosis.

Nuptial pads represent a key sexually dimorphic trait in anurans, with male Rana chensinensis exhibiting these structures on the Finger I of forelimbs. However, the specialized mechanisms underlying their functional adaptation remain poorly understood. This study aimed to characterize the structural and molecular features of these nuptial pads by using Finger Ⅲ as a control. Histological staining and transcriptome analysis were conducted to compare glandular morphology, collagen fiber characteristics, and gene expression profiles between the two fingers. Results revealed significantly larger specialized mucous glands (SMGs) and thicker collagen fibers in the nuptial pads, along with downregulation of KRT-related genes and catalase (CAT), and upregulation of genes associated with small heat-shock proteins (sHSPs), melanogenesis, and motor proteins. These findings suggest that nuptial pads achieve functional adaptation through synergistic structural (mechanical support via enlarged glands and thickened fibers) and molecular (metabolic regulation and secretory optimization) mechanisms. Such adaptations align with the demands of successful amplexus and ovulation stimulation, associated with collectively ensuring reproductive success in anurans.

Aging is associated with progressive impairment of hepatic adaptive capacity, including mitochondrial dysfunction, oxidative stress, and disruption of proteostasis. Persistent activation of the unfolded protein response (UPR), particularly under chronic endoplasmic reticulum (ER) stress, has been implicated in age-related liver vulnerability even in the absence of metabolic disease. This study evaluated the effects of time-restricted feeding (TRF), either alone or in combination with exercise training, on age-associated hepatic alterations in adult (6-month-old) and aged (24-month-old) male C57BL/6 J mice maintained on a standard diet. TRF consisted of a 12-h fasting/feeding cycle applied 5 days per week for 8 weeks, and exercise comprised alternating aerobic and resistance sessions (1 h/day). Aged mice exhibited sustained activation of PERK-associated ER stress signaling, particularly along the PERK-CHOP axis, accompanied by mild inflammatory cell infiltration, increased collagen deposition, and elevated MASLD score, despite the absence of increased adiposity or impaired glucose tolerance. TRF alone partially attenuated PERK-associated ER stress signaling and histological alterations; however, only the combined intervention significantly reduced upstream PERK phosphorylation. Persistent elevation of CHOP across aged groups suggests that downstream PERK-associated stress signaling were not fully reversible. Collectively, these findings demonstrate that hepatic aging under non-obesogenic conditions is characterized by sustained PERK-associated ER stress signaling accompanied by mild inflammatory cell infiltration and increased collagen deposition, and that coordinated nutritional and exercise strategies can modulate stress-responsive pathways associated with hepatic resilience during aging.

One of the most efficient and promising viral vectors for the treatment of central nervous system (CNS) and peripheral nervous system (PNS)-related diseases is adeno-associated virals (AAVs), which in recent years, researchers have succeeded in substantially managing and mitigating these disorders by leveraging innovative therapeutic mechanisms at the cellular and molecular levels, which are widely utilized due to their favorable characteristics, including a robust safety profile, the ability to cross the blood-brain barrier, and differential tropisms for distinct types of neuronal cells. Clinically, AAVs have high specificity and have therefore become a dynamic and useful tool for the gene therapy of a variety of neurological diseases. Many researchers around the world have been able to study a variety of neurological diseases in vitro cellular models, in vivo animal models, and human clinical studies using these vectors. Given these characteristics, the overall aim of the present review is to investigate the role of AAV vectors carrying coding and non-coding RNAs in the treatment of a variety of neurological disorders, to study the relationship between these vectors and cellular signaling pathways, and to examine its clinical applications in various neurological diseases.

Epithelial-mesenchymal transition (EMT) is increasingly recognized as a dynamic process of epithelial-mesenchymal plasticity (EMP) rather than a binary switch between epithelial and mesenchymal phenotypes. This plasticity enables tumor cells to acquire invasive, metastatic, stem-like, and therapy-resistant characteristics that contribute to cancer progression and poor clinical outcomes. Despite extensive investigation, significant challenges remain in understanding the functional significance of partial EMT states, identifying robust biomarkers that capture EMT heterogeneity, and translating EMT-targeted interventions into clinical practice. This review critically examines the molecular mechanisms governing EMT, including transcriptional regulators, signaling pathways, epigenetic modifications, and tumor microenvironmental influences. Particular emphasis is placed on hybrid epithelial/mesenchymal states, which have emerged as key drivers of metastatic fitness and therapeutic resistance. We further evaluate established and emerging EMT biomarkers, including E-cadherin, N-cadherin, vimentin, circulating tumor cells, and exosome-derived signatures, highlighting their opportunities and limitations in clinical applications. Finally, we discuss current therapeutic strategies targeting EMT-associated pathways and explain why many promising preclinical approaches have faced translational barriers. By integrating mechanistic, biomarker, and therapeutic perspectives, this review proposes that understanding and targeting epithelial-mesenchymal plasticity, rather than EMT alone, may provide a more effective framework for precision oncology and metastasis prevention.

Persistent organic pollutants such as organochlorine pesticides (OCPs) have been implicated in obesity and metabolic dysfunction. As they bioaccumulate in adipose tissue, OCPs may act as obesogens, but in vivo evidence for aldrin, endosulfan and lindane is limited. This study investigated whether subacute exposure to aldrin, endosulfan and lindane elicits obesogenic effects in male rats by examining weight gain, serum biochemistry and metabolic and inflammatory gene expression in white adipose tissue (WAT) and brown adipose tissue (BAT). Thirty-two adult male Sprague-Dawley rats were randomly assigned to receive corn oil (control) or oral aldrin, endosulfan or lindane (1 mg/kg every other day for 28 days). Body weight was recorded weekly. Serum liver enzymes and lipid profile were determined. In WAT and BAT, we quantified mRNA expression of thermogenic and adipogenic markers (Pparg, Ucp1, Ucp3, Fndc5), and the pro- and anti-inflammatory cytokines (Il-1b, Tnf-α, Il-10, Il-4). Lindane caused significant weight gain from the first week (p < 0.0001), whereas aldrin and endosulfan increased body weight only at week four (p < 0.05). Endosulfan-treated rats had reduced total and high-density lipoprotein cholesterol (p < 0.05) without changes in low-density lipoprotein cholesterol, very-low-density lipoprotein cholesterol, or triglycerides. In WAT, Pparg, Ucp1, and Fndc5 transcripts were unchanged, while Ucp3 tended to decrease in the endosulfan group (p = 0.056). In BAT, endosulfan increased Ucp1 and decreased Fndc5 levels (p < 0.05), with no changes in Pparg or Ucp3. Endosulfan suppressed Il-1b, Tnf-α, and Il-10 (p < 0.05) transcripts in both WAT and BAT, and all pesticides reduced Il-4 expression in BAT (p < 0.05). Overall, short-term exposure to aldrin, endosulfan or lindane interfered with metabolic and inflammatory pathways in a compound-specific manner, with lindane causing early weight gain and endosulfan eliciting complex effects on lipid metabolism and adipose gene expression. We suggest that even short-term exposure to OCPs may affect metabolic regulation in male rats, with each chemical exerting distinct effects.

The metabolic determinants that distinguish Crohn's disease (CD) from ulcerative colitis (UC) and their roles in driving immune dysregulation remain elusive. We aimed to identify systemic metabolic signatures of inflammatory bowel disease (IBD) and elucidate the mechanistic link between metabolite availability and regulatory T cell (Treg) homeostasis. We performed untargeted LC-MS-based metabolomics on plasma from patients with CD (n = 17), UC (n = 13), and healthy controls (n = 15). Pathway enrichment and topology analysis were used to identify perturbed biological processes. The functional role of identified metabolites was validated using primary murine naive CD4+ T cell differentiation assays, Seahorse XF metabolic flux analysis, and siRNA-mediated knockdown. The therapeutic relevance was assessed in a DSS-induced murine model of colitis. Metabolomic profiling identified 175 significant differential metabolites, revealing a profound systemic depletion of fatty acids and a specific deficit in the fatty acid oxidation (FAO) pathway in CD patients. Palmitic acid and butyrate emerged as robust diagnostic biomarkers (AUC = 0.877-0.975). In vitro, butyrate dose-dependently promoted iTreg differentiation by increasing the oxygen consumption rate and maximal respiratory capacity. This effect was dependent on carnitine palmitoyltransferase 1A (CPT1A), as pharmacological or genetic inhibition of CPT1A-mediated FAO abolished butyrate's pro-differentiation capacity. In vivo, the ability of butyrate to attenuate colitis and promote colonic iTreg accumulation was significantly reversed by the CPT1 inhibitor etomoxir. CPT1A-mediated fatty acid oxidation is a critical metabolic pivot through which butyrate signals to maintain immune tolerance. The systemic loss of FAO intermediates in CD patients directly contributes to impaired iTreg development, identifying the butyrate-CPT1A axis as a potential target for precision metabolic intervention.

Alzheimer's disease (AD) is an irreversible neurodegenerative disorder and the major cause of dementia globally. One of the major barriers to curing AD is the lack of experimental models that fully recapitulate the complexity of human AD pathology. Evidence supports the notion that neuroinflammation is a key pathological contributing factor of AD. Therefore, various external stimuli are used to recapitulate the AD-like pathology, i.e., LPS, scopolamine, and aluminum chloride. Although PHA is reported for its pro-inflammatory activity, using PHA to recapitulate the AD-like pathology is relatively new. The current study was designed to evaluate the PHA potential to recapitulate the AD-like pathology in primary neuronal culture as well as in vivo AD modeling. Since LPS is widely used, in the current study, it was used to compare it with the experimental findings using PHA. Confirmation of AD-like pathology was carried out through gene and protein expression in the in vitro study, while in vivo results were verified through behavioral and histopathological assays. It was observed that PHA mimics the AD-like pathology at the cellular level by aggregating, shrinking, and disrupting synaptic plasticity of primary neuronal cells. An elevated level of BACE1 and amyloid-β and hyperphosphorylated tau protein expression, along with reduced levels of the ADAM10 gene, evidently proves that PHA induces the AD-like pathology. Disrupted cognitive and memory patterns confirm the initiation of AD pathogenesis in the in vivo study, which is further confirmed by observing the amyloid plaques stained by Congo red (CR) dye. Based on these findings, it is concluded that PHA is a potent inducer to recapitulate neuroinflammatory-mediated AD-like pathology both in vitro and in vivo.

The thermodynamic logic underlying hemoglobin's cooperative binding and reversible conformational transitions offers a powerful conceptual model for reimagining polymer design in neurodegenerative medicine. In this review perspective, we outline a unified thermodynamic framework that connects molecular energetics, polymer science, and pathological protein aggregation. We discuss how hemoglobin's allosteric adaptability, enthalpy-entropy compensation, and redox responsiveness can inspire polymers capable of sensing and reshaping the free-energy landscapes that govern amyloid formation. Drawing on evidence from protein thermodynamics, polymer chemistry, and neurobiological systems, we propose design principles for adaptive, hemoglobin-inspired polymers that act as artificial chaperones, materials capable of modulating aggregation equilibria, restoring proteostatic balance, and integrating diagnostic and therapeutic functions. This article defines an emerging field at the intersection of thermodynamic polymer science and neurodegeneration, where materials are not passive carriers but active regulators of molecular energy landscapes.

A mutation of the ADAM20 gene was described in an infertile patient whose spermatozoa were observed accumulating in the perivitelline space of oocytes. To decipher its role in the fertilization process, we produced a mouse model knock-out (KO) for the Adam20 gene. Surprisingly, Adam20-KO males showed an in vivo fertility similar to their wild-type littermates and had normal sperm counts and morphology. However, they presented a reduced fertility in in vitro fertilization assays. To explore the hypothesis that the two flanking genes Adam25 and Adam39 could compensate the lack of Adam20 given their very high homology, we produced a triple KO deleting these three neighbor genes simultaneously. The phenotype was strictly identical to that of the single gene deletion. Therefore, ADAM20, ADAM25, and ADAM39 are not essential proteins for fertilization in mice but they seem to participate modestly to the process. Adam20-KO males are fertile in vivo but have a reduced fertility in vitro. Males deleted for three neighbor and very homologous genes (Adam20, Adam25, and Adam39) have the same phenotype as the simple Adam20-KO. These genes are dispensable for normal fertility but contribute modestly to fertilization.

Apoptosis, as the predominant form of programmed cell death, plays a crucial role in maintaining tissue homeostasis. Apoptotic bodies (ABs), which are extracellular vesicles released during the terminal phase of apoptosis, were initially characterized as functionless cellular debris. However, they are now recognized for mediating intercellular communication via biomolecule delivery in both physiological and pathological states. Owing to their diverse biological properties, ABs have emerged as promising therapeutic candidates for treating a wide range of diseases, including cancer, cardiovascular and bone diseases, neurodegenerative disorders, diabetes, and infectious/inflammatory conditions, and have demonstrated potential in promoting tissue regeneration. Furthermore, ABs present in bodily fluids carry abundant disease-related information and offer a dynamic representation of patients' physiological states, rendering them valuable tools for diagnosis and prognosis. This review consolidates progress in ABs research, delineates their pathophysiological roles, and assesses challenges and opportunities for their clinical application.

The ARHGEF (Rho guanine nucleotide exchange factor) family acts as a key upstream regulator of Rho GTPases, including RhoA, Rac1, and Cdc42. It plays critical roles in cytoskeletal remodeling, cell migration, vascular tone regulation, and inflammatory responses. Recent studies have shown that dysregulated ARHGEF activity contributes to cardiovascular diseases such as atherosclerosis, hypertension, cardiac hypertrophy, fibrosis, and myocardial ischemia/reperfusion injury. However, the functions of ARHGEFs in specific cell types and their overall signaling mechanisms in the cardiovascular system remain poorly understood. This review provides a comprehensive framework of current progress in understanding the molecular mechanisms underlying ARHGEF-mediated regulation in endothelial dysfunction, vascular remodeling, myocardial fibrosis, and ischemia/reperfusion injury. It also highlights potential pharmacological strategies that target the ARHGEF and Rho GTPase signaling axis for precise therapeutic intervention. By integrating molecular, cellular, and translational research, this work offers a conceptual framework for ARHGEF-driven signaling in cardiovascular diseases, and offers new insights for future drug development.

Myokines are defined as a class of bioactive molecules-including metabolites, peptides, and proteins-released from skeletal muscle cells and promoted by exercise. Recently, myokine-mediated muscle-organ crosstalk has sparked increased interest. Skeletal muscle secretes hundreds of myokines in an autocrine, paracrine, or endocrine manner, mediating the crosstalk between skeletal muscle and skeletal muscle itself, bone, fat, liver, and so on to regulate the physiological state of multiple organs, exerting a wide range of benefits of exercise. The relationship between muscle and myokines may be better understood as an exercise-responsive regulatory framework involving skeletal muscle and myokines, although direct evidence for an integrated bidirectional feedback network remains incomplete. In the network, myokines regulate skeletal muscle metabolism and remodeling through autocrine/paracrine actions, while exercise-induced muscle-derived signals may also contribute to long-lasting systemic adaptations involving immune mobilization, redox homeostasis, gut microbiota-related metabolic remodeling, and gut-muscle-bone crosstalk. Therefore, the distinct feature of skeletal muscle is not the mere presence of bidirectional signaling but its large mass, fiber-type metabolic heterogeneity, and direct responsiveness to repeated mechanical stimuli during exercise. On the one hand, as a source of myokines, the stimulation of skeletal muscle affects the production of myokines through various signaling pathways. On the other hand, myokines affect skeletal muscle function, such as glucose absorption, fatty acid oxidation, skeletal muscle mass, and muscle fiber type transformation. What's more, some myokines are muscle fiber type-specific, meaning that the expression of myokines may be influenced by muscle fiber type. Based on these, this review aims to summarize current evidence for exercise-responsive interactions between skeletal muscle and myokines and to identify where feedback regulation remains hypothetical.

Deep mining of the molecular mechanisms underlying skin aging is critical for the development of novel therapeutic targets. The aim of this study was to explore potential key signatures and underlying mechanisms in skin aging. Bioinformatics tools were used to evaluate the differentially expressed genes (DEGs), perform enrichment analysis, and construct protein-protein interaction networks on datasets. The hub genes were identified using Cytoscape software and then confirmed by qRT-PCR and Western Blot in vitro. We focused on the possible roles of GRWD1. GRWD1 expression were suppressed in senescence model of HDF. After GRWD1 overexpression by lentivirus in primary human dermal Fibroblasts (HDF), transcriptome sequencing (RNA-seq) was performed. GRWD1 has influence on the inflammation and immune responses. IFN signaling pathway genes significant increase in senescence HDF. When GRWD1 overexpressed, decreased expression of IFN pathway is validated. Besides, age-related markers p16, IL-6, and IL-8 expression decreased and LaminB expression also increased. It was also proved after GRWD1 knockdown. GRWD1 was identified as a novel signature in skin aging. GRWD1 directly interact with IRF3. GRWD1 overexpression can down regulate IFN signaling pathway key genes expression, further diminish IFN aging-inducing effects. GRWD1 alleviates skin inflammatory responses by suppressing IRF3-mediated IFN pathway activation. Our findings suggest that GRWD1 may serve as a molecular biomarker and further as a potential therapeutic target for skin aging.