Philadelphia chromosome-positive (Ph+) leukemia is among the most aggressive forms of leukemia and arises from the Philadelphia chromosome (Bcr-Abl fusion gene) abnormality. This disease is associated with a poor prognosis in adults and resistance to traditional treatment. Thus, the identification of novel therapeutic targets and drugs for Ph+ leukemia remains crucial. S-phase kinase-associated protein 2 (Skp2), a key component of the SCF ubiquitin ligase complex, is aberrantly overexpressed in various malignancies and therefore represents a potential target for anticancer therapy. This study aimed to elucidate the therapeutic mechanism of action of AAA-237, a novel Skp2 inhibitor, in Ph + leukaemia. Cellular responses to AAA-237 were evaluated using cell viability, EdU incorporation, and colony formation assays. Pyroptosis was observed by scanning electron microscopy (SEM), immunofluorescence, lactate dehydrogenase (LDH) detection, ELISA, and Western blotting. Phospholipid metabolism was evaluated through metabolomic profiling and RNA sequencing. Mechanistic findings were validated using Western blotting and quantitative real-time PCR. Mitochondrial and endoplasmic reticulum damage were evaluated using Ca2+, ATP, TMRM, and mitochondrial reactive oxygen species (ROS) assays. In vivo efficacy and safety of AAA-237 were examined in a xenograft model. The results showed that AAA-237 targeted Skp2 and inhibited Bcr-Abl phosphorylation and ubiquitination, suppressing the proliferation of Ph+ leukemia cells in a concentration-dependent manner at the cellular level. AAA-237 induced both pyroptosis and apoptosis through caspase-dependent mechanisms. Treatment with AAA-237 inhibited the PI3K/AKT signaling pathway and disrupted phospholipid metabolism, resulting in dysregulated phospholipid metabolism. AAA-237 also increased PACS2 expression, remodeled mitochondria-associated endoplasmic reticulum membranes (MAMs), and induced mitochondrial dysfunction and endoplasmic reticulum damage. In vivo, AAA-237 effectively suppressed leukemia progression. These results show that AAA-237, a novel Skp2 inhibitor, inhibits the proliferation of Ph+ leukemia cells by reducing Bcr-Abl phosphorylation and ubiquitination. AAA-237 further induces caspase-dependent pyroptosis and apoptosis by disrupting phospholipid metabolism and impairing MAM homeostasis. These results provide preliminary experimental evidence supporting the development of targeted candidate therapeutics for Ph+ leukemia.

Endogenous cytoplasmic DNA (cytoDNA) is increasingly recognized as a mediator of tissue dysfunction and disease progression during aging. As a major cytosolic DNA-sensing pathway, the cyclic GMP-AMP synthase-stimulator of interferon genes (cGAS-STING) pathway can translate aging-associated cytoDNA accumulation into innate immune and inflammatory programs. This review summarizes the evolutionary and signaling features of the cGAS-STING cascade and critically discusses its crosstalk with aging-associated intracellular molecular threats, including nuclear genomic and chromatin stress, mitochondrial dysfunction, oxidative-metabolic stress, and defective clearance of nucleic acids or damaged organelles. We further synthesize evidence linking dysregulated cGAS-STING activation to inflammatory remodeling, senescence-associated changes, cell injury, fibrosis, and tissue dysfunction, while highlighting the context-dependent roles of this pathway across physiological aging and ARDs. Finally, we discuss the therapeutic potential and limitations of cGAS-STING modulation, emphasizing that successful translation will require context-defined therapeutic windows, tissue- and cell-specific targeting, subcellular compartmentalization, and long-term safety assessment.

Alzheimer's disease (AD) progression involves complex molecular mechanisms underlying neuronal dysfunction. While emerging evidence on long noncoding RNAs (lncRNAs) is accumulating, the relevance of nuclear noncoding RNAs (ncRNAs) to neurodegenerative diseases remains poorly understood. Small Cajal body-specific RNA 13 (scaRNA13) is a nuclear ncRNA implicated in RNA species regulation, which remains insufficiently characterized in neuronal systems and AD pathogenesis. Here, we performed integrative analyses of human postmortem brain transcriptomes and AD mouse models to examine scaRNA13 expression across disease stages, sex, and brain regions. RNA-seq and proteomic analyses were used to assess scaRNA13-associated changes in gene expression, splicing, and RNA-protein interactions. Functional assays in neuronal cells were conducted to evaluate the effects of scaRNA13 perturbation on RNA processing, protein synthesis, and tau-related pathology. scaRNA13 was aberrantly upregulated in AD patient brains with a pronounced elevation observed in female patients at advanced stages. Perturbation of scaRNA13 altered splicing patterns and global translational capacity, accompanied by altered tau aggregation- and phosphorylation-related phenotypes in neuronal cell systems. These findings support scaRNA13 as an AD-associated nuclear ncRNA candidate and suggest that scaRNA13 perturbation is associated with changes in RNA processing, translational regulation, and tau-related cellular phenotypes in neuronal cell systems.

Tumor organoids preserve the cellular heterogeneity and structural complexity of native tumors, providing robust platforms for mechanistic studies, preclinical drug testing, and translational oncology research. However, their predictive performance is not an intrinsic property of organoids. It is shaped by platform design, including culture format, matrix composition, medium formulation, and the extent of multicellular reconstruction. This review examines how these design variables influence model fidelity, reproducibility, tumor microenvironment reconstruction, and drug-response interpretation. We compare major culture formats and matrix systems, discuss strategies for incorporating stromal and immune components, and evaluate current and emerging uses of AI-assisted analysis for organoid-derived phenotypic data. Overall, this review highlights how integrated tumor organoid design can strengthen pharmacological modeling and oncology translation.

DNA double-strand breaks (DSBs) represent a major threat to genomic integrity. DSBs are primarily repaired through homologous recombination (HR) and non-homologous end-joining (NHEJ). The Ku complex, composed of XRCC5 and XRCC6, initiates NHEJ by binding to the DNA ends in DSBs. Our previous study demonstrated that FOXL2 regulates Ku availability through acetylation-dependent interaction; however, whether related mechanisms operate across other FOX family proteins and how these can be regulated in response to genotoxic stress remain unclear.In this study, we demonstrated that multiple FOX family members interact with XRCC5/6 and acetylation-dependently suppress NHEJ. GCN5 (KAT2A) promoted the acetylation of a conserved lysine residue within the forkhead domain, thereby enhancing FOX-Ku association and limiting Ku availability. In response to DNA damage, SIRT1 underwent SUMOylation at lysine 734, thereby promoting its nuclear accumulation through KPNA2/KPNA3-dependent import. Nuclear SIRT1 acted as a deacetylase and reversed FOX acetylation, thereby reducing FOX-Ku interaction and permitting NHEJ activation.The results of this study define a DNA damage-responsive GCN5-SIRT1 regulatory axis that modulates FOX acetylation, FOX-Ku interaction, and NHEJ activity. Our study extends FOXL2-centered findings and supports a shared, but context-dependent, DNA repair regulation-supporting mechanism involving multiple FOX proteins.

Skeletal muscle, once regarded solely as a contractile tissue, is now recognized as a dynamic endocrine organ that secretes exercise-induced myokines-bioactive peptides with autocrine, paracrine, and endocrine functions. These myokines coordinate systemic energy homeostasis by regulating glucose and lipid metabolism, mitochondrial function, inflammation, and interorgan communication. Building on our previous review published in 2018, this review synthesizes major advances in exercise-induced myokines within an evidence-based framework considering mechanistic support and translational relevance. We highlight both well-established and emerging myokines, including interleukin-6 (IL-6), irisin, myostatin, growth differentiation factor 11 (GDF11), IL-15, brain-derived neurotrophic factor (BDNF), meteorin-like (METRNL), secreted protein acidic and rich in cysteine (SPARC), fibroblast growth factor 21 (FGF21), β-aminoisobutyric acid (BAIBA), leukemia inhibitory factor (LIF), apelin, and musclin, and discuss their roles across major target tissues including skeletal muscle, liver, adipose tissue, and bone. We also summarize natural and synthetic compounds reported to modulate myokine expression, secretion, or activity, and discuss the opportunities and current limitations of targeting myokine pathways. Although several myokine axes show therapeutic promise, the current literature indicates substantial heterogeneity in causal evidence, receptor or target certainty, and translational readiness. These insights support a more selective view of myokines as biologically heterogeneous mediators of muscle-organ crosstalk and provide a framework for mechanism-based therapeutic development in metabolic disease.

Extracellular heat shock protein 90 alpha (eHsp90α) has been implicated in promoting cancer cell invasion and metastasis. However, the role of eHsp90α in the tumor microenvironment remains poorly understood. In the present study, we provide the first evidence that eHsp90α aggravates the invasiveness of cancer-associated fibroblasts (CAFs) through low-density lipoprotein receptor-related protein 1 (LRP1) and consequent upregulation of epithelial-mesenchymal transition (EMT)-associated transcription factor expression. We observed that eHsp90α induces CAFs invasiveness. However, silencing of Twist, Snail and Slug expressions significantly attenuated eHsp90α-induced CAFs invasiveness. In addition, transfecting CAFs with LRP1 short interfering RNA (siRNA) markedly inhibited eHsp90α-induced CAFs invasion and EMT-associated transcription factor expression. Intriguingly, we observed that the epidermal growth factor receptor (EGFR)/AKT signaling is located downstream of LRP1 and governs eHsp90α-induced Twist and Snail but not Slug expression required for CAFs invasion. Moreover, we demonstrated that these EMT-associated transcription factors of CAFs are indispensable for breast cancer cell invasion. Collectively, the present data reveal that eHsp90α increases CAFs invasiveness through LRP1 and subsequent EMT-associated transcription factor expression to promote breast cancer invasion, presenting potential therapeutic targets to impede breast cancer progression.

Liver organoids are emerging as human-relevant three-dimensional in vitro systems for chemical safety assessment, however, their application value depends on their ability to generate decision-ready evidence within clearly defined context-of-use (CoU). Here, we review the biological fidelity, quality assessment, exposure characterization, and validation requirements of liver organoids across three CoUs: (1) hazard screening and identification, (2) mechanistic elucidation of hazard, and (3) quantitative risk assessment support. Within this CoU framework, we review recent advances in liver organoids and their applications to chemical safety assessment, highlighting models that already provide useful evidence and those where implementation remains partial or inconsistent. Based on this review, we discuss the major barriers to broader regulatory adoption, which include insufficient biological implementation, limited reproducibility, insufficient exposure characterization, and a lack of harmonized performance criteria and standardization frameworks. By evaluating liver organoids in relation to clearly defined decision questions rather than generalized technological promise, this review clarifies their current contributions, explains why their contributions remain CoU-dependent, and outlines key priorities for developing decision-ready liver organoid platforms for next-generation chemical safety assessment.

Sepsis-associated encephalopathy (SAE), the most common neurological complication of sepsis with high mortality, lacks effective treatments. Although microglia play a significant role in SAE pathogenesis, the mechanisms of M2 microglial exosomes (M2-EXOs) and their microRNAs (miRNAs) remain unclear. The present study demonstrated that both M2 microglial conditioned media (M2-CM) and M2-EXO promote the phenotypic transformation of M1 microglia toward the M2 phenotype. In M2-EXO, among the differentially expressed miRNAs, miR-1949 exhibited the most significant difference. The dual-luciferase assay showed that miR-1949 could bind to the 3'UTR of Dickkopf-related protein 1 (DKK1). Further experiments confirmed that M2-EXO-miR-1949 mimic mediated microglial phenotypic transformation by inhibiting DKK1 and activating the Wnt/β-catenin pathway. Notably, a single injection of M2-EXO (10 μg/mouse, i.c.v.) or M2-EXO-miR-1949 mimic (0.1 nmol/mouse, i.c.v.) significantly reduced the mouse sepsis score (MSS), decreased cortical neural cell damage in SAE mice induced by lipopolysaccharide (LPS), inhibiting abnormal microglial activation and alleviating neuroinflammation. Meanwhile, M2-EXO-miR-1949 mimic markedly inhibited DKK1 and activated the Wnt/β-catenin pathway in vivo. What's more, the effects of M2-EXO-miR-1949 mimic were reversed by the Wnt/β-catenin pathway inhibitor XAV939. In contrast, the effect of M2-EXO-miR-1949 inhibitor (0.1 nmol/mouse, i.c.v.) was opposite to that of M2-EXO-miR-1949 mimic. Furthermore, co-transfection of the M2-EXO-miR-1949 mimic and inhibitor mutually neutralized their opposing effects, resulting in no significant changes in the expression of inflammatory and pathway-related markers, thereby confirming the functional specificity of exosomal miR-1949. In summary, the present study indicates that M2-EXO, especially its contained miR-1949, ameliorates LPS-induced SAE in mice through inhibiting DKK1 to activate the Wnt/β-catenin pathway, thereby leading to microglial phenotypic transformation and neuroinflammation inhibition. This study reveals a novel mechanism by which M2-EXO and miRNA ameliorate SAE, providing a theoretical basis for further studies.

Chronic inflammation drives the pathogenesis of diverse disorders, yet current anti-inflammatory strategies often lack mechanistic selectivity. Glycyrrhizic acid (GL), a major triterpenoid saponin from Glycyrrhiza species, exhibits broad anti-inflammatory effects, but its mechanism has been viewed as non-specific. In this review, we propose an integrated framework positioning high-mobility group box 1 (HMGB1) as one major molecular hub for GL's action. Mechanistically, our molecular docking analysis suggests that GL may interact with the Box B domain of HMGB1 andmay potentially interfere with its engagement with TLR4 and RAGE, thereby orchestrating a cascade inhibition of downstream NF-κB, MAPK, and NLRP3 inflammasome pathways. Beyond this core signaling axis, we systematically analyze how GL context-dependently modulates macrophage polarization, monocyte-derived macrophage differentiation, and Th17/Treg balance. We further synthesize its effects across liver, autoimmune, metabolic, oncologic, neurological, and renal diseases, revealing shared mechanisms and disease-specific adaptations. Finally, we critically evaluate translational bottlenecks-including isomer-specific bioavailability, the pseudoaldosteronism risk, and formulation challenges-and propose strategies to overcome them. By shifting from a descriptive summary to a mechanism-centered, integrative analysis, this review provides a conceptual framework for developing GL as a multitarget anti-inflammatory agent, while acknowledging its limitations including low oral bioavailability, microbiota-dependent metabolism, pseudoaldosteronism risk, and limited clinical evidence.

Using a [1]H-NMR-guided isolation approach, twelve novel hasubanan alkaloids, bishernsubanins A-D (1-4) and hernsubanines G-N (5-12), along with eight known monomeric analogues (13-20), were isolated from Stephania japonica var. discolor. Notably, compounds 1-4 represent a new class of homologous dimeric hasubanan alkaloids. Their structures were elucidated by comprehensive spectroscopic data analysis, including NMR, ECD calculations, and single crystal X-ray diffraction. Their biosynthetic pathways are proposed to involve redox reactions, free radical coupling, isomerization, and condensation reactions. All isolates were evaluated for anti-neuroinflammatory activity in vitro. Alkaloids 1, 3, 4, 7, and 11-13 exhibited better inhibitory effects on nitric oxide production in lipopolysaccharide (LPS) induced BV-2 microglial cells than minocycline. Among them, the new compound bishernsubanin A (1) could inhibit Iba-1 expression, and suppress the release of inflammatory factors in LPS stimulated BV-2 microglial cells. Molecular docking, immunofluorescence, and drug affinity responsive target stability (DARTS) studies indicated that compound 1 modulates toll-like receptor 4 to exert its anti-neuroinflammatory effect, positioning it as a promising naturally derived candidate for neuroinflammatory inhibition.

Previous studies have shown that Quzhou Fructus Aurantii extract (QFAEE) ameliorates lipopolysaccharide (LPS)-induced acute lung injury (ALI) by modulating the Stimulator of interferon genes (STING) pathway. Nevertheless, the mechanisms underlying the anti-inflammatory effects of nobiletin (Nob), the primary bioactive component of QFAEE, have not been clearly elucidated. The present study aimed to investigate the protective effects of Nob on a mouse model of LPS-induced ALI and elucidate its underlying mechanism. The pulmonary inflammatory response in mice was quantified by performing fluorescence-based RT‒qPCR, bronchoalveolar lavage fluid (BALF) cell counting, and hematoxylin‒eosin (HE) staining. Transcriptomic profiling was performed to explore the mechanisms by which Nob ameliorates LPS-induced ALI. The anti-inflammatory mechanisms were further elucidated using western blotting, immunohistochemical staining, cellular thermal shift assays (CETSAs), and microscale thermophoresis. Functional validation was conducted using the STING agonist vadimezan (DMXAA), the STING inhibitor SN-001, and STING-knockout (STING-KO) mice. Nobiletin-loaded liposomes (Nob-Lipo) significantly attenuated LPS-induced pulmonary inflammation in mice. The results of the transcriptomic analysis suggested that this protective effect was associated with the modulation of the type I interferon pathway. In RAW264.7 and THP-1 cells, pretreatment with Nob effectively attenuated the interferon regulatory factor 3 (IRF3)-mediated type I interferon response and nuclear factor kappa-B (NF-κB)-dependent proinflammatory cytokine expression following DMXAA-induced STING pathway activation. Furthermore, the STING inhibitor SN‑001 markedly suppressed LPS‑induced inflammatory responses, as evidenced by significantly reduced levels of proinflammatory cytokines and interferon‑stimulated genes, and effectively blocked the nuclear translocation of NF‑κB and IRF3. Notably, after inhibitor treatment, Nob no longer exerted its previously observed anti‑inflammatory effects, further supporting the critical regulatory role of STING in LPS‑mediated inflammation and indicating that the anti‑inflammatory activity of Nob is primarily dependent on the STING pathway. Moreover, in STING‑KO mice with LPS‑induced ALI, the loss of the protective effect of Nob further confirmed that its anti‑inflammatory activity depends on STING, highlighting the essential role of STING signaling in mediating its protective effects. In conclusion, the protective effect of Nob on an LPS-induced ALI model is mediated primarily through the modulation of STING signaling.

The widespread dissemination and serious clinical consequences of hypervirulent Klebsiella pneumoniae (hvKP), an encapsuled gram-negative pathogenic bacterium characterized by its specific virulence factor hypermucoviscosity (HMV), is becoming a concerning global public threat. In contrast to the common health care-associated infection of classical K. pneumoniae (cKp), hvKP is more virulent and capable of causing community-acquired invasive infections in healthy individuals, which mainly relies on the increased production of capsular polysaccharides (CPS) termed hypercapsule, and capsule-associated HMV. The indispensable role of capsule in mediating immune evasion, including resisting phagocytic engulfment and the killing of serum and host-derived antibacterial peptides, and other pathogenesis, renders it an attractive target for drug development against K. pneumoniae. Here, we identified the natural phenylpropanoid compound cinnamic acid (CA) as an effective inhibitor of K. pneumoniae capsule. The significant inhibition effect of CA on capsule biosynthesis was verified by both biochemical analysis and microscopic observation, and such drug action functioned in multiple K. pneumoniae strains. Mechanistically, CA hindered capsule biosynthesis by increasing bacterial carbon metabolism and consequently energy metabolism. Accordingly, hypercapsule-conferred hypermucoviscosity phenotype of hvKP was prominently impeded by CA. As a result, the cellular adherence and phagocytosis ratio, as well as serum killing and antibacterial peptide activity, were all improved by the inhibitor. In vivo, CA treatment significantly protected Galleria mellonella and mice from lethal hvKP infection. In conclusion, this study demonstrates that CA is a potent K. pneumoniae capsule inhibitor, which provides an alternative therapeutic strategy and active compound for K. pneumoniae infections.

Lactylation, as a novel post-translational modification, plays a key role in gene transcription and protein expression, serving as an important link between cellular metabolism and functional regulation. Ferroptosis is a regulatory form of cell death characterized by iron-dependent lipid peroxidation, and its abnormal regulation is closely associated with the onset and progression of various diseases, including tumors, degenerative diseases, and ischemia-reperfusion injury. Recent studies have shown that lactylation can regulate ferroptosis-related pathways and key targets through multiple mechanisms, regulate ferroptosis and thereby influence disease progression. This review first comprehensively summarizes the mechanisms of lactylation and ferroptosis, then systematically reviews the regulatory mechanisms and roles of lactylation in ferroptosis, and finally, we summarize the role of lactylation-regulated ferroptosis in various diseases and its potential as a therapeutic target. The aim is to deepen our understanding of the regulatory mechanisms and roles of lactylation in ferroptosis and to lay the foundation for the development of precision treatment strategies for related diseases.

Mitochondrial DNA copy number (mtDNA-CN) is a critical marker of mitochondrial health and plays a key role in cellular bioenergetics. Alterations in mtDNA-CN have been associated with aging, metabolic disorders and neurodegenerative diseases. Recent studies have revealed that various plant-derived extracts, as well as the secondary metabolites they produce, known as phytochemicals, can modulate mtDNA-CN through mechanisms including the regulation of mitochondrial biogenesis, oxidative stress, and mtDNA repair. This review examines plant-derived extracts and phytochemical compounds from a wide range of plant species- including Ginkgo biloba, Crocus sativus, Curcumin and many others- able to modulate mtDNA dynamics, scavenging oxygen free radicals and improving antioxidant defense systems.