Large-scale randomized trials have found that multivitamin-multimineral (MVM) supplements and cocoa flavanols may benefit several age-related chronic conditions among older adults, but it remains unclear whether these two supplements directly slow the biological aging process. This prespecified ancillary study evaluated the 2-year effect of a daily MVM (Centrum Silver) and cocoa extract (500 mg cocoa flavanols per day, including 80 mg (-)-epicatechin) on five DNA methylation measures of biological aging (PCHannum, PCHorvath, PCPhenoAge, PCGrimAge and DunedinPACE) among 958 participants (482 women and 476 men) in the COcoa Supplement and Multivitamin Outcomes Study (COSMOS). Compared with placebo, daily MVM supplementation modestly reduced the rate of increase of second-generation epigenetic clocks, with a between-group difference in yearly change of -0.113 years (95% confidence interval (CI) -0.205 to -0.020; P = 0.017) for PCGrimAge and -0.214 years (-0.410 to -0.019; P = 0.032) for PCPhenoAge. MVM had a stronger effect on PCGrimAge among those with accelerated biological aging at baseline (-0.236 [-0.380 to -0.091]) compared with those with normal or decelerated biological aging (-0.013 [-0.130 to 0.104]; P = 0.018 for interaction). Cocoa extract did not have an effect on the five epigenetic clocks tested. Although the statistically significant but small effects of daily MVM supplementation on slowing biological aging are encouraging, additional studies are needed to determine the clinical relevance of daily MVM supplementation on epigenetic clocks and whether such effects can help explain the beneficial effects of MVM supplementation on aging-related chronic conditions.

Polycystic ovary syndrome (PCOS) is associated with ovarian granulosa cell dysfunction. Ferroptosis, a regulated cell death driven by lipid peroxidation, represents a novel pathological mechanism. Hypermethylation of the glutathione peroxidase 4 (GPX4) promoter may contribute to its suppression. While glucagon-like peptide-1 receptor agonists (GLP-1 RAs) improve metabolic features of PCOS, their direct effects on ovarian ferroptosis and the underlying epigenetic mechanisms are unclear. To explore the therapeutic potential of GLP-1RAs across PCOS phenotypes, we employed a hyperandrogenism-induced rat model and a letrozole plus high-fat diet mouse model, treating them with exenatide or semaglutide, respectively. Phenotypic assessment included estrous cycle monitoring, ovarian histology, and serum hormone profiling. Ferroptosis was evaluated using a multi-parametric approach, including iron deposition (Perls' staining), lipid peroxidation (MDA), redox status (GSH/GSSG), ultrastructural analysis (TEM), and expression of key regulators. The methylation status of the GPX4 promoter was analyzed by methylation-specific PCR (MSP) and bisulfite sequencing (BSP), alongside the expression of related epigenetic modifiers (DNMTs, TET1). In vitro studies using DHT-stimulated primary granulosa cells further validated the semaglutide effects. GLP-1 RA exenatide alleviated the polycystic ovarian morphology in rats with PCOS, semaglutide treatment not only alleviated PCOS phenotypes but also reversed ovarian ferroptosis markers, restored GPX4 expression, and reduced the GPX4 promoter hypermethylation and DNMTs levels, with efficacy comparable to 5-azacytidine. In vitro, semaglutide corrected DHT-induced GPX4 hypermethylation and ferroptosis in granulosa cells. This study demonstrates that semaglutide alleviates PCOS phenotypes and reverses ovarian granulosa cell ferroptosis. These beneficial effects may be related to the alleviation of GPX4 promoter hypermethylation. Our findings extend the therapeutic rationale for semaglutide in PCOS beyond metabolic benefits, suggesting potential direct ovarian protection via epigenetic modulation.

Multiple myeloma (MM) is an incurable plasma cell neoplasm that is highly reliant on endoplasmic reticulum-associated degradation (ERAD) to maintain protein homeostasis. Disrupting ERAD has been proposed as a therapeutic strategy to overcome proteasome inhibitor resistance; however, the identification of novel inhibitors has been limited. To address this, we conducted a cell-based high-throughput screen using the FDA repurposing library and identified omaveloxolone (RTA408) as a potent ERAD inhibitor that selectively impairs the degradation of ER luminal and membrane substrates, without affecting the degradation of key cytosolic proteins that are implicated in disease relapse. Surprisingly, although ER stress response pathways are activated after ERAD inhibition in MM, we find that apoptosis is mediated by altered lipid raft organization, leading to aberrant activation of the death-inducing signaling complex (DISC) and caspase 8 in the extrinsic apoptotic pathway. Notably, ERAD inhibition by RTA408 is cytotoxic to primary malignant plasma cells, including those resistant to proteasome inhibitors, and demonstrates in vivo anti-myeloma activity. Our findings establish a novel ERAD inhibitor, which is a valuable tool to dissect ERAD biology, and provide pre-clinical evidence for RTA408 as a therapeutic agent in MM.

The treatment of colorectal cancer (CRC) remains challenging due to chemotherapy resistance and genetic heterogeneity. Indole-3-lactic acid (ILA), a tryptophan metabolite derived from gut microbiota, exhibits promising anti-inflammatory and anticancer properties; however, its specific molecular targets and regulatory mechanisms in CRC remain poorly understood. In this study, we combined network pharmacology and machine learning with molecular docking to identify candidate targets and pathways for ILA in CRC. We identified 39 ILA-CRC common targets, ultimately identifying four hub genes through the intersection of machine learning models. Validation in independent GEO datasets confirmed significant differential expression of these genes in CRC tissues. Functional enrichment analyses linked these genes to the PPAR, PI3K-AKT, and IL-17 signaling pathways, and gene set enrichment analysis further implicated ascorbate and aldarate metabolism, DNA replication, and fatty acid metabolism. Immune infiltration analysis indicated associations between hub gene expression and immune cell populations, including mast cells, neutrophils, and macrophages, suggesting potential involvement in the tumor immune microenvironment. Molecular docking supported favorable binding of ILA to all four hub proteins, and 100-ns molecular dynamics simulations specifically validated the dynamic stability of the ILA-HMOX1 complex. In conclusion, these results highlight EPHA2, HMOX1, MMP3, and PARP1 as candidate targets and suggest that ILA may influence CRC-related signaling, metabolic programs, and immune contexture, providing a theoretical foundation for developing gut microbiota-derived metabolites as novel anticancer strategies.

Glucocorticoids (GCs) are steroid hormones that bind to the glucocorticoid receptor (GR) as ligands to initiate systemic anti-inflammatory effects. GCs are commonly administered alongside chemotherapy to reduce treatment-related side effects in breast cancer patients. However, GC administration has been shown to promote metastasis in breast cancer. In this study, we used quantitative mass-spectrometry-based approaches to analyze proteome and phosphoproteome of three breast cancer cell lines following treatment of a clinically approved synthetic GC, dexamethasone (Dex). By comparing MCF7, MDA-MB-231, and MDA-MB-436 cells, we suggest that the level of GR significantly affects Dex-mediated responses. Additionally, we identify noncanonical transcription factors (TFs) and kinases that are regulated by GR in different cell lines. Together, our data present Dex-induced protein modulations and modifications involving several TFs and kinases that regulate cytoskeletal remodeling and migration in breast cancer cell lines. These findings highlight the need for careful consideration of GC use in breast cancer therapy and identify potential molecular targets for mitigating adverse effects.

MsWIP3, a C2H2-type zinc finger transcription factor in alfalfa, plays a key role in enhancing saline-alkali stress tolerance by regulating stress responses and reducing oxidative damage. Saline-alkali stress severely restricts plant growth and yield worldwide, particularly in the Songnen Plain of northeastern China, where alkaline salts with high pH values, mainly NaHCO₃, are a major constraint on agriculture and forage production. C2H2-type zinc finger proteins (C2H2-ZFPs) are important transcription factors involved in plant responses to multiple environmental stresses. However, their roles in alfalfa (Medicago sativa) adaptation to saline-alkali stress remain poorly understood. We integrated transcriptome (RNA-seq) analysis of alfalfa under NaHCO₃ treatment with Weighted Gene Co-expression Network Analysis (WGCNA) to identify stress-responsive transcription factors. MsWIP3, a WIP subfamily member, contains four conserved zinc finger domains and the characteristic "WIP" motif. Subcellular localization and transcriptional activity assays were performed, and the function of MsWIP3 was evaluated through heterologous expression in yeast and Nicotiana benthamiana. MsWIP3 localized to the nucleus and exhibited transcriptional repression activity. Overexpression of MsWIP3 suppressed the growth of yeast and tobacco but significantly enhanced NaHCO₃ tolerance in transgenic tobacco, as evidenced by reduced PSII photoinhibition and lower oxidative damage from reactive oxygen species. Our findings indicate that MsWIP3 plays a role in the saline-alkali stress response of alfalfa. This study contributes to our understanding of the molecular mechanisms of stress adaptation and identifies a potential target for the development of salt-alkali tolerant alfalfa varieties.

Cell surface receptors play vital roles in cancer growth and metastasis. Integrin αvβ3 is overexpressed in various cancer cells and interacts with different growth factors to stimulate cancer progression. Thyroid hormone binds to αvβ3 to activate signal transduction and cell proliferation. However, thyroxine (T4) deaminated analogue, tetraiodothyronine (tetrac), competes for the binding on integrin and inhibits cancer cell growth and metastasis. The current study investigated the pathogenic role of integrin αvβ3 and the potential of a novel therapeutic strategy targeted to integrin αvβ3. Pathogenetic studies of clinical samples revealed integrin αvβ3 cross-talked with EGFR and downstream signal transduction networks affected by thyroid hormone and EGF related to the progression of cholangiocarcinoma malignancy. Thyroxine and EGF stimulated PD-Ligand 1 (PD-L1) expression and cancer growth in cholangiocarcinoma. The thyroxine-induced PD-L1 accumulated in the nuclei and colocalized with p300. Alternatively, EGF increased cytosolic PD-L1 and nuclear accumulation of β-catenin. Targeting integrin αvβ3 with lipo-tetrac and its Dox-derivative induced anti-proliferation in vitro and in the xenografted animal model. Our research provides a fundamental understanding of the therapeutic role of integrin αvβ3 and the potential therapeutic approach in cholangiocarcinoma treatment.

Triple-negative breast cancer (TNBC) is an aggressive subtype lacking targetable hormone receptors, making conventional chemotherapy the primary treatment option, despite its associated toxicity and potential for drug resistance. Melatonin, a natural hormone with anticancer and immunomodulatory properties, has shown promise in multiple cancers; however, its role in TNBC remains unclear.

Prostate cancer (PCa) is the most prevalent malignancy in men and often correlates with distant metastasis in its advanced stages. The study aimed to investigate the effects of Ugonin J, a natural compound isolated from Helminthostachys zeylanica, on PCa metastasis.

B-cell lymphoma 6 (BCL6) is a transcriptional repressor whose overexpression is closely linked to the progression of diffuse large B-cell lymphoma (DLBCL), making it a promising therapeutic target. This study aims to identify a novel small molecule, synthesized via proteolysis-targeting chimeras (PROTACs), capable of degrading BCL6, thereby inhibiting DLBCL growth and providing a foundation for future preclinical studies.

Tamoxifen is a key drug that provides endocrine therapy for estrogen receptor (ER) α-positive breast cancer; however, resistance remains a significant clinical challenge. This study aims to investigate the molecular mechanisms of tamoxifen resistance in ERα-positive breast cancer, with particular focus on the role of SET Domain Containing 1A (SETD1A)-driven forkhead box A2 (FOXA2) as a key regulator of this resistance.

Clear cell renal cell carcinoma (ccRCC) is predominantly treated with anti-angiogenic therapies (AATs), such as sunitinib and axitinib. While these therapies initially improve outcomes, resistance frequently emerges, limiting long-term efficacy. Understanding the molecular mechanisms underlying AAT resistance is essential to optimize treatment strategies.

PD-L1 is one of the most critical immune checkpoint proteins, inhibiting T-cell immune responses by binding to PD-1. This study aims to validate that allicin can regulate PD-L1 expression through the IL-6/JAK2/STAT3 pathway, thereby inhibiting immune evasion in osteosarcoma.

Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive fibrosing interstitial lung disease without any effective treatment. Berberine (BBR), a botanical alkaloid, possesses extensive biological activities and has significant therapeutic value in various diseases. However, the effect and potential mechanisms of BBR on pulmonary fibrosis remain elusive. In vivo, BBR was administered by gavage following intratracheal instillation of bleomycin (BLM) in a mouse model from Day 1 to Day 20. In vitro, Human Lung Fibroblast (HLF) and A549 cell lines were used to explore the effects of BBR on transforming growth factor β1 (TGF-β1) treated cells. Both cell lines were transfected with a lentivirus carrying TGF-β receptor 2 (TGFBR2) knockdown genes, and the autophagy inhibitor chloroquine (CQ) and PI3K inhibitor LY294002 were employed to investigate the underlying effects of BBR on TGF-β signaling and autophagy in pulmonary fibrosis. BBR administration attenuates pulmonary inflammation and fibrosis of BLM-induced mice in vivo. Analogously, BBR treatment significantly alleviates matrix collagen deposition and reduces the expression of fibrotic markers in TGF-β1-treated human lung fibroblasts (HLF) and alveolar epithelial cell (A549) in vitro. Mechanistically, we found that BBR downregulates the expression of TGFBR2 and suppresses TGF-β/Smad2/3 signaling in vivo and in vitro. Furthermore, BBR inhibits the activation of the PI3K/AKT/mTOR pathway and autophagy, then downregulates the expression of pro-fibrotic genes. The effect of BBR on pulmonary fibrosis was further verified using both TGF-β1-treated HLF and A549 cells with the addition of the inhibitors of PI3K, LY294002, and autophagy, CQ in vitro, respectively. Our study suggests that BBR can inhibit pulmonary fibrosis by down-regulating the expression of TGFBR2, attenuating TGF-β/Smad2/3 signal, and activating autophagy through phosphorylation of PI3K/AKT/mTOR.

Vibrio alginolyticus is a widespread marine pathogen responsible for major disease outbreaks in aquaculture and zoonotic infections in humans. Its pathogenicity is tightly regulated by quorum sensing and driven by biofilm formation, motility, and multiple virulence factors. The organism's high adaptability and virulence complicate traditional antibiotic treatments, especially amid rising antibiotic resistance. Although thymol, a natural monoterpenoid, is known for its broad-spectrum bactericidal activity, its effects on the survival and virulence of V. alginolyticus have not been previously investigated. In this study, we evaluated the inhibitory potential of thymol against V. alginolyticus. Experimental results demonstrated that thymol significantly reduced the survival of both planktonic and mature biofilm cells, induced reactive oxygen species generation, and disrupted membrane integrity. Furthermore, thymol at sub-inhibitory concentrations markedly impaired quorum sensing regulated biofilm formation and motility. Global transcriptomic analysis revealed that thymol exerts a multifaceted inhibitory mechanism by downregulating key genes involved in antioxidant defense, membrane biosynthesis, energy metabolism, ABC transporter function, quorum sensing, biofilm formation, and motility. In addition, computational studies revealed strong binding affinities between thymol and the upstream key quorum sensing regulators LuxS and LuxU, with hydrogen bonds formed at key active site residues, further supporting its anti-virulence potential. To our knowledge, this is the first study demonstrating that thymol acts as a potent inhibitor of V. alginolyticus survival and virulence. These findings highlight thymol as a promising natural agent for managing V. alginolyticus associated infections in aquaculture systems.

In tomato, the 25 AHL family members were classifi ed into three subfamilies. SlAHL5 and SlAHL25 belonged to CladeB, interacted and enhanced salt and osmotic stress tolerence of transgenic Arabidopsis. AT-hook motif nuclear-localized (AHL) proteins participate in plant growth, development, and response to abiotic stress. Their functions in the resistance to salt and osmotic stresses are largely unknown in tomato. Here, a total of 25 AHL genes in the tomato (Solanum lycopersicum) genome were identified. Phylogenetic and gene structure analyses indicated that they were classified into two clades and three subfamilies. Synteny relationship analysis demonstrated that all paralogous SlAHL pairs evolved under purifying selection. Promoter structure analysis revealed that many stress-related and phytohormone-related cis-acting elements existed. Gene expression pattern assays indicated that they had significantly different expressions in various organs, and most of them were up-regulated by high salinity and/or osmotic stress. Yeast two-hybrid (Y2H) assays demonstrated that SlAHL5 and SlAHL25, two nucleus-localized members in Clade B, interacted with each other to form a heterodimer, with SlAHL5 having self-activation activity, which is absent in SlAHL25. Constitutive expression of either SlAHL5 or SlAHL25 increased the resistance of transgenic plants to both salt and osmotic stresses, as revealed by the promoted primary root growth and biomass production, the relatively higher chlorophyll and proline content, and the enhanced catalase (CAT) and peroxide dismutase (POD) activity under both stress conditions. Our study on SlAHL genes under different stress conditions reported here provides a basis for further functional analysis of SlAHL genes, as well as for the development of new breeding strategies to improve resistance to multiple abiotic stresses in tomato.

Cardiac arrest (CA) remains a major public health challenge with high incidence and mortality. Post-cardiac arrest brain injury (PCABI) is the primary determinant of poor neurological outcomes and survival. Although curcumin (Cur) exhibits neuroprotective effects in multiple cerebral injury models, its precise pharmacological mechanisms in PCABI remain incompletely understood.

Hepatocellular carcinoma (HCC) is a primary malignant tumour that impacts patients' quality of life. Currently, clinical experience from The First Affiliated Hospital of Guangzhou University of Chinese Medicine suggests that Jianpi Jiedu Xiaozheng Fang (JPJDXZF) demonstrates promising efficacy in the treatment of HCC. We aimed to explore the mechanisms of JPJDXZF in HCC based on network pharmacology. The components and their relevant targets of JPJDXZF were identified using databases such as SymMap, TCMID, TCMSP, and TCM-ID. Following ADME screening, 1443 active components of JPJDXZF were identified, and 435 corresponding drug targets were predicted using the SwissTargetPrediction database. Subsequently, prognosis-related differentially expressed genes (DEGs) associated with HCC were analyzed using TCGA and GTEx datasets, and a gene expression matrix was derived. Key genes involved in HCC regulation were identified, and functional analyses were performed. Furthermore, we explored the regulatory effects of JPJDXZF at the cellular, organoid, and animal levels. We identified 18 intersecting genes between HCC prognosis-related genes and JPJDXZF-target genes. Venn diagram analysis successfully identified BIRC5 and CYP2E1 as two potential targets for JPJDXZF in treating HCC. Pathway enrichment analysis indicated that the core targets of JPJDXZF were enriched in multiple signalling pathways, including the Hippo pathway, in which BIRC5 is involved as a downstream regulatory gene. In in vitro experiments, JPJDXZF-containing serum significantly reduced the viability and migration of HepG2 and MHCC97-H cells, leading to a decrease in organoid diameter and ATP activity in HCC organoids. In in vivo experiments, tumours in nude mice treated with JPJDXZF exhibited reduced volume and weight, along with decreased expression of BIRC5 and Hippo pathway effectors YAP and TAZ. At the mechanistic level, JPJDXZF treatment was associated with altered Hippo pathway-related signalling, accompanied by reduced YAP/TAZ activity and changes in BIRC5 expression, together with effects on HCC cell proliferation and apoptosis. In addition, siMST1/2 interference and EMT inhibitor-1 treatment partially attenuated the effects of JPJDXZF on cell viability, migration, and apoptosis. JPJDXZF regulates BIRC5 expression in association with Hippo pathway activity in HCC. In vitro, in vivo, and molecular mechanism analyses support JPJDXZF as a potential therapeutic strategy for HCC by modulating key proteins in the Hippo pathway, thus affecting HCC cell proliferation, apoptosis, and migration.

Histone deacetylase 6 (HDAC6) has emerged as a promising therapeutic target in cancer due to its immunomodulatory effects. While its prognostic significance remains debated, we demonstrate that HDAC6 loss significantly impairs myeloid leukemia progression in vivo, despite having no functional impact on leukemia cell proliferation in vitro. Global proteome and secretome profiling of HDAC6-knockout (KO) cells revealed upregulation of several immune-related modulators, including RNase T2, a tumor suppressor known to modulate the tumor microenvironment. Notably, RNase T2 upregulation upon HDAC6 loss was observed in myeloid leukemia cells but not in lymphoblastic leukemia cells. Moreover, pharmacological inhibition of HDAC6 recapitulated this phenotype, leading to RNase T2 upregulation in myeloid leukemia cells. ATAC-seq revealed increased chromatin accessibility of RNase T2 following HDAC6 loss, highlighting a functionally epigenetic regulatory contribution. Further functional assays conducted in an immunocompetent setting, both ex vivo and in vivo, demonstrated that HDAC6 inhibition sensitized murine myeloid leukemia cells to broad CD8+ T cell activation as evidenced by increased TNFα and CD107a expression. Consistently, in a syngeneic murine model, HDAC6 inhibition restricted the growth of myeloid leukemia cells. Moreover, an extended drug screening analysis identified Cytarabine and Clofarabine as significantly synergizing with HDAC6 inhibitor (Ricolinostat) in myeloid leukemia cell lines and in patient-derived xenograft (PDX) cells, while showing limited synergy in lymphoid leukemia cell lines, PDX, or healthy control cells. These findings suggest that HDAC6 represents a promising therapeutic target in myeloid lineage-derived leukemia cells by simultaneously enhancing immune activation and increasing chemosensitivity.

Silicosis remains a critical occupational health concern worldwide, lacking effective treatments due to unclear mechanisms. In this study, we investigated the citrullinated proteomic profile and its effects in mice exposed to silica. Our findings demonstrated elevated levels of citrullinated peptides and citrullinated vimentin (Cit-Vim) in silicotic mice and silica-treated macrophages, regulated by peptidylarginine deiminase (PADI2). Unlike vimentin, Cit-Vim amplified the production of tumor necrosis factor-α (TNF-α), Interleukin-6 (IL-6), and IL-1β in silica-treated macrophages through interaction with Toll-like receptor 4 (TLR4) signaling. RNA sequencing revealed that early growth response protein 1 (EGR1) is a target of PADI2, with Cit-Vim inducing lung inflammation via EGR1 signaling. Pharmacological inhibition or genetic knockout of Padi2 attenuated silica-induced lung inflammation and fibrosis. These findings suggest that targeting PADI2 may represent a novel therapeutic strategy of silicosis.