Gambogic acid inhibits tumor growth and induces HMGB1-mediated pyroptosis in AML models in vitro and in vivo.

Structural characterization and anticancer enhancement of selenized squid ink glycosaminoglycan nanoparticles.

Role of the GAPDH-Mediated Glycolysis-Senescence Axis in Osteoarthritis Chondrocytes and Its Modulation by Semaglutide.

Regenerative endodontics focuses on the restoration of the pulp-dentine complex by promoting odontogenic differentiation of stem cells of the apical papilla (SCAPs). Although SSUH2 has been implicated in developmental processes and dentine dysplasia type I (DD-I), the specific role of SSUH2 in regulating SCAPs differentiation remains unclear. In this study, we identified SSUH2 as a novel biomarker for SCAPs odontogenic differentiation and revealed its critical regulatory mechanism. Immunohistochemical staining of mouse mandibular first molar sections and immunofluorescence staining of human dental pulp tissues were performed to investigate the expression characteristics of SSUH2 during root development. SCAPs were treated with siRNA-mediated knockdown or lentivirus-mediated overexpression of SSUH2, followed by assessment of proliferation and odontogenic differentiation. Subcutaneous implantation of hydrogel-scaffold-root fragments loaded with SSUH2-overexpressing SCAPs in nude mice was performed to evaluate invivo odontogenic capacity. Mechanistically, transcriptome sequencing and bioinformatic analysis identified downstream signalling pathways. Co-immunoprecipitation, nuclear-cytoplasmic fractionation, and immunofluorescence confirmed the direct interaction between SSUH2 and FOXM1, promoting FOXM1 nuclear translocation and subsequent transcriptional upregulation of PDK1. Moreover, transmission electron microscopy, mitochondrial membrane potential assays and ROS detection collectively demonstrated that SSUH2 regulates mitochondrial function in SCAPs via the FOXM1/PDK1 axis. During the development of the first molar in the mouse's lower jaw, SSUH2 is enriched in the root tip papilla region and it is also enriched in the root tip papilla region of humans. SSUH2 positively regulated the odontogenic differentiation of SCAPs invitro. Invivo transplantation models showed that SSUH2 overexpression enhanced the odontogenic differentiation capacity of SCAPs and promoted dentine-pulp-like structure formation. Transcriptomic and functional analyses indicated that SSUH2 maintained mitochondrial function. SSUH2 directly interacted with FOXM1, promoted its nuclear translocation, and upregulated PDK1 expression. FOXM1 silencing abrogated SSUH2-mediated enhancement of both mitochondrial function and odontogenic differentiation in SCAPs. Our study is the first to reveal that SSUH2 promotes the odontogenic differentiation of SCAPs through the FOXM1/PDK1 axis by regulating mitochondrial function. These findings suggest that SSUH2 could serve as a potential therapeutic target for enhancing the odontogenic differentiation of SCAPs and provide a novel strategic direction for pulp-dentine complex regeneration.

Background: Colistin (Col) resistance and heteroresistance in extensively drug-resistant (XDR) Acinetobacter baumannii severely limit therapeutic options. We investigated the activity and mechanism of human albumin nanoparticles (haNPs) as colistin potentiators against genetically characterized clinical isolates. Methods: Sixteen clinical isolates were analyzed. Col MICs were determined by broth microdilution, and heteroresistance by population analysis profiling. Potentiation of Col activity was assessed using both Col-loaded haNPs (Col/haNPs) and free Col co-administered with empty haNPs, alongside the proton motive force (PMF) uncoupler carbonyl cyanide 3-chlorophenylhydrazone (CCCP). Assays included checkerboard synergy (FICI), membrane potential analysis (DiOC2(3)), intracellular Col quantification (UPLC-MS/MS), zeta potential measurements, transmission electron microscopy (TEM), protein leakage, and ROS detection. Results: Heteroresistance was detected in 9/16 isolates. Col/haNPs reduced Col MICs by 4-64-fold in resistant strains and shifted MICs to ≤2 mg/L in most heteroresistant isolates. Empty haNPs displayed no intrinsic antibacterial activity yet selectively potentiated Col, with strong synergy (FICI down to 0.035). Membrane depolarization and increased intracellular Col accumulation under haNP-treated conditions paralleled the effects of CCCP, indicating that haNPs elicit a CCCP-like functional response. These findings are compatible with perturbation of membrane energetics and possible downstream effects on PMF-dependent transport processes. TEM and surface charge analyses supported direct nanoparticle-envelope interaction and progressive membrane disruption. Conclusions: haNPs enhance Col activity across genetically diverse A. baumannii isolates, with particularly strong effects in heteroresistant strains. The combined effects of PMF modulation, increased intracellular drug availability, and envelope interaction provide a mechanistic rationale for the use of albumin-based nanoparticles, either as Col carriers or in combination with free drug, to overcome Col resistance and heteroresistance.

BackgroundColorectal cancer (CRC) continues to be a major cause of cancer-related mortality worldwide, creating an urgent need to develop novel therapeutic agents that are both highly effective and minimally toxic. Gamabufotalin (GA), a bioactive bufadienolide compound, has shown promising antitumor potential, including against CRC; however, its precise mechanisms of action in CRC remain incompletely understood.ObjectivesThe aim of this study was to investigate the therapeutic effect of GA on CRC and explore the underlying molecular mechanisms.MethodsA combination of in vitro and in vivo approaches was employed, including cell viability assays, colony formation, EdU incorporation, cell cycle and apoptosis analyses by flow cytometry, ROS and mitochondrial membrane potential detection, as well as integrated transcriptomic and proteomic profiling. The in vivo antitumor efficacy was further validated in a nude mouse xenograft model.ResultsGamabufotalin exhibited potent, dose-dependent anti-CRC activity in vitro (IC50: 24-30 nM) with high selectivity over normal cells. It triggered mitochondrial apoptosis via ROS generation and arrested the cell cycle, suppressing DNA synthesis. Integrated omics revealed TP53I3/PIG3 upregulation and RFC3/NUCKS1 downregulation as key mechanisms. Critically, these effects converged to suppress tumor growth in vivo without systemic toxicity.ConclusionsGamabufotalin selectively and potently inhibits CRC through dual mechanisms: Inducing ROS-mediated apoptosis and suppressing proliferation. With a favorable therapeutic index and defined molecular targets, GA represents a promising candidate for CRC therapy.

Background/Objectives: The frequent use of antibiotics has led to increasing drug resistance in Aeromonas hydrophila; therefore, there is an urgent need to develop novel antimicrobial agents to prevent and control bacterial diseases in aquaculture. Allicin E (ALE) is derived from garlic (Allium sativum L.), a plant extensively used in traditional medicine for treating infections. This study aimed to evaluate the potential of ALE against A. hydrophila, a major aquaculture pathogen, by investigating its antibacterial efficacy, mechanisms of action, and in vivo protective effects. Methods: The minimum inhibitory and bactericidal concentrations (MIC/MBC) were determined by broth microdilution. Antibacterial mechanisms were investigated through ROS detection, electron microscopy, fluorescent staining, and content leakage measurement. In vivo efficacy was evaluated in Carassius auratus gibelio by monitoring survival rates and bacterial loads, analyzing immune and antioxidant biomarkers, and histopathological analysis after A. hydrophila challenge. Results: ALE exhibited potent antibacterial activity (MIC = MBC = 8 μg/mL), achieving complete bacterial elimination within 1 h and showing a low resistance propensity. Mechanistically, ALE induced ROS accumulation, causing oxidative damage that disrupted membrane integrity and facilitated the leakage of cellular contents. In vivo, ALE significantly enhanced fish survival, reduced bacterial loads, modulated inflammatory cytokines, boosted antioxidant enzyme activities (SOD and CAT), and alleviated tissue damage. Conclusions: ALE possesses potent in vitro antibacterial activity and exerts an inhibitory effect on bacteria-induced inflammatory responses, effectively combating A. hydrophila through a multi-target mechanism and enhancing host resistance.

SSD: Targeting inflammasome and oxidative stress as a therapeutic strategy in inflammatory diseases.

Polysaccharide from Tetrastigma hemsleyanum Diels et Gilg attenuates Poly(I:C)-induced acute lung injury by preserving epithelial barrier integrity and modulating STING/TBK1-NF-κB and IRF1-STAT1 signaling.

Direct antibacterial activity of allicin and restoration of carbapenem susceptibility by inhibiting New Delhi metallo-β-lactamase.

By analyzing the single-cell RNA-Seq libraries we established of OA joints during exercise therapy, we found cartilage intermediate zone might participate in early OA exercise therapy. Early OA rat model was established by 4-week anterior cruciate ligament transection (ACLT). The radiomics was used to evaluate the relative damaged and undamaged area in OA patients’ cartilage. We overexpressed and knocked down CILP in early OA chondrocyte to explore its potential mechanism. The quantitative proteomics was used to examine the protein profiles of the CILP-treated chondorcyte. The Yeast One-Hybrid Assay, Co-Immunoprecipitation (Co-IP), Nrf2 cytosol-nuclei fractionation and ubiquitination assay were used to investigate the potential mechanism in CILP intervention. Western blot, ROS, JC-1, Ferrous ion, MDA and GSH detection, transmission electron microscopy (TEM) were used to explored the therapeutic effect of CILP on OA. Moderate exercise up-regulates CILP in the articular cartilage intermediate zone. CILP recovers the ratio of type II / I collagen, Sox9, α-SMA expression and competitively bind to Keap1 protein and reduce the stability of Keap1-Nrf2 dimer, thereby reducing the degree of Nrf2 ubiquitination and promoting Nrf2 nuclear translocation. Nrf2 nuclear translocation activated SLC7A11, HO-1, GPX4 and SOD-1 expression, then decreased the MDA contents, but increased GSH content, which ultimately inhibited chondrocytes ferroptosis and promoted hyalinization of fibrocartilage. Exercise induced cartilage intermediate zone and CILP-Keap1-Nrf2 axis inhibits hyaline cartilage fibrosis and chondrocyte ferroptosis to alleviate early osteoarthritis.

Oral Delivery of siSIRPα via Yeast-Derived β-Glucan Particles Enhances Macrophage-Mediated Antitumor Immunity by Blocking the CD47-SIRPα Axis.

Transfer RNA-derived small RNAs (tsRNAs) represent a novel class of non-coding RNAs increasingly implicated in cardiovascular regulation. However, their roles in sepsis-induced cardiomyopathy (SICM) remain largely undefined. This study aimed to investigate the function and underlying mechanism of 5'tiRNA-32-LysCTT-11-a highly upregulated tsRNA in SICM-in modulating myocardial injury. A murine model of sepsis was established via cecal ligation and puncture (CLP), and myocardial injury was assessed by serum CK-MB/LDH levels, histology, and cardiac function via echocardiography. In vitro, H9C2 cardiomyocytes were exposed to conditioned media (CM) from lipopolysaccharide (LPS)-stimulated macrophages. The expression of 5'tiRNA-32-LysCTT-11 was measured by qRT-PCR. Functional assays including CCK-8, LDH release, PI staining, JC-1, ATP, ROS detection, and MitoTracker staining were performed. Necroptosis was evaluated via MLKL phosphorylation; Mitochondria-associated endoplasmic reticulum membranes (MAMs) formation was assessed by dual-label immunofluorescence and Pacs2 expression. Bioinformatics analysis identified Mitofusin 2 (Mfn2) as a putative target, validated by Western blot, mRNA stability assay (Actinomycin D), and rescue experiments. 5'tiRNA-32-LysCTT-11 was significantly upregulated in SICM. In vivo, its overexpression improved cardiac function and reduced injury biomarkers. In vitro, 5'tiRNA-32-LysCTT-11 mimics preserved mitochondrial integrity, reduced ROS and ATP depletion, suppressed MAM formation and necroptosis. Inhibitor transfection produced opposite effects. Mechanistically, 5'tiRNA-32-LysCTT-11 enhanced Mfn2 mRNA stability and protein expression. Silencing Mfn2 abrogated the protective effects, confirming its central role in the tsRNA's action. 5'tiRNA-32-LysCTT-11 exerts cardioprotective effects during sepsis by stabilizing Mfn2 mRNA, preserving mitochondrial function, limiting MAMs formation, and suppressing necroptosis. These findings uncover a novel regulatory mechanism and suggest 5'tiRNA-32-LysCTT-11 as a promising therapeutic target in SICM.

Atherosclerosis is a chronic inflammatory disease characterized by lipid-driven immune dysregulation. Argininosuccinate synthase 1 (ASS1) has been implicated in macrophage inflammation, yet its precise mechanistic role in foam cell-mediated vascular injury during atherosclerosis remains unclear. This study investigates whether ASS1 promotes disease progression via the NLRP3/IL-33/ST2 axis. An in vitro foam cell model was established using phorbol 12-myristate 13-acetate (PMA)-differentiated U937 macrophages treated with oxidized low-density lipoprotein (ox-LDL). The role of ASS1 was assessed via knockdown (si-ASS1) and overexpression (ASS1 overexpression) plasmids. Co-culture systems with human umbilical vein endothelial cells (HUVECs) and human aortic vascular smooth muscle cells (HAVSMCs) were used to evaluate endothelial apoptosis and VSMC proliferation/migration. In vivo, atherosclerosis was induced in apolipoprotein E‑deficient (ApoE)-deficient mice via a 12-week high-fat diet, and ASS1 expression was modulated using AAV9 vectors. Molecular analyses included ROS detection, enzyme-linked immunosorbent assay (ELISA), qPCR, western blot, and immunofluorescence. Plaque burden was assessed via Oil Red O staining. Ox-LDL treatment significantly upregulated ASS1 expression in U937-derived foam cells. ASS1 overexpression enhanced intracellular ROS production, NLRP3 inflammasome activation, STAT3 phosphorylation, and IL-33 secretion. These effects were reversed by ASS1 knockdown. Rescue experiments demonstrated that STAT3 is required for ASS1-mediated NLRP3 activation and IL-33 upregulation. ASS1 altered IL-33 receptor ST2 signaling by increasing the soluble decoy isoform (sST2) and decreasing the membrane-bound signaling isoform (ST2L). In co-culture, ASS1-overexpressing foam cells promoted HUVEC apoptosis (via mitochondrial pathway) and HAVSMC proliferation, migration, and dedifferentiation. NLRP3 overexpression alone mimicked the pro-inflammatory effects of ASS1 and reversed the anti-inflammatory effects of ASS1 knockdown. In vivo, ASS1 knockdown in ApoE-/- mice reduced plaque lipid deposition, serum levels of IL-33 and IL-1β, and vascular expression of NLRP3 and p-STAT3, while ASS1 overexpression exacerbated these parameters. ASS1 drives atherosclerosis by activating the STAT3/NLRP3 inflammasome axis, shifting the IL-33/ST2 balance toward a pro-inflammatory state, and amplifying foam cell-mediated endothelial injury and smooth muscle cell dysfunction. Targeting ASS1 may offer a novel therapeutic strategy for inflammatory vascular disease.

Primary open-angle glaucoma (POAG) is the leading cause of irreversible blindness. Regrettably, the roles of ferroptosis-related (FR) genes in POAG remain elusive. Five GEO data sets and a series of experimentations in vitro were used for bioinformatic exploration and biological validation. Using multiple machine learning algorithms, four critical FR genes in POAG progression were screened. The clinical value and biological function of NOX1 were comprehensively analyzed using bioinformatic approaches. A POAG in vitro model was constructed using H2O2 treatment. NOX1 effects on the viability of retinal ganglion cells (RGCs) and ferroptosis process were determined through CCK8, EdU, ROS detection, and transmission electron microscopy. Its upstream transcriptional mechanisms were determined through dual luciferase assays, and chromatin immunoprecipitation (ChIP). NOX1 was identified as the critical FR gene in POAG progression and served as an effective diagnostic biomarker. High-NOX1 expression was tightly associated with increased infiltration levels of multiple subtypes of T cells, such as T cells CD8 and T cells CD4. However, the enrichments of eight metabolic gene sets did not differ between the POAG samples with high- and low-NOX1 expression groups. Silencing NOX1 maintained RGC survival and inhibited the ferroptosis process. Mechanistically, STAT3 upregulated NOX1 by binding its promoter region that was located at the 429th to 419th bases upstream of the NOX1 transcriptional start site. NOX1 overexpression reversed the inhibitory effects of silencing STAT3 on RGC survival and the ferroptosis process. NOX1 was a good biomarker for characterizing POAG and promoted POAG progression through STAT3-mediated transcriptionally activation.

Sepsis is a life-threatening condition characterized by uncontrolled inflammation, oxidative stress, and aberrant cell death. Although the STING pathway is well established as a central mediator of innate immunity, its functions beyond interferon signaling remain largely undefined. Here, we investigated a noncanonical STING–ERO1–PARP1 axis that drives oxidative damage and Parthanatos during sepsis. A combination of in vivo and in vitro approaches was employed to dissect the STING–ERO1–PARP1 signaling cascade. A sepsis model was established via cecal ligation and puncture, and various genetic knockout mice (STING⁻/⁻, cGAS⁻/⁻, and IFNAR1⁻/⁻) were used to assess pathway specificity. In vitro studies in RAW264.7 and iBMDM cells involved STING agonist stimulation combined with genetic knockouts (STING⁻/⁻, TBK1⁻/⁻), targeted inhibitors (PJ34, EN460, NAC), and siRNA transfection to dissect signaling mechanisms. Molecular and cellular responses were evaluated by immunoblotting, immunofluorescence, ELISA, TUNEL staining, and ROS detection. Cell death, cytokine expression, and tissue injury were quantified via standard assays. we demonstrated that the STING signaling axis drives parthanatos and intestinal injury through endoplasmic reticulum (ER) oxidative stress. Mechanistically, STING directly interacts with ER oxidoreductase 1 (ERO1), inducing oxidative stress and cytosolic ROS accumulation, which triggers DNA damage, PARP1 hyperactivation, and excessive PARylation. This cascade promotes the nuclear translocation of apoptosis-inducing factor (AIF) and Parthanatos. Notably, this mechanism occurred independently of the canonical cGAS–TBK1–interferon axis. Genetic deletion or pharmacological inhibition of STING, ERO1, or PARP1 significantly reduces ROS levels, PARylation, and intestinal injury. We identified a novel STING–ERO1–PARP1 signaling cascade that links innate immune sensing to redox dysregulation and cell death, providing new therapeutic targets for mitigating organ injury in sepsis.

Monocrotaline induces liver injury via TRX1-ASK1-JNK Axis-mediated mitochondrial damage in hepatocytes.

From traditional tonic herb to molecular therapy: The natural peptide G-6-Y from Eleutherococcus sessiliflorus protects against drug-induced liver injury.

TRIM59 alleviates neuronal ferroptosis and promotes functional recovery after spinal cord injury by mediating ubiquitination and degradation of ANXA2.

Septic cardiomyopathy (SCM) is a common and life-threatening complication of severe sepsis, with high mortality due to unclear underlying mechanisms. CXCL4, a key pro-inflammatory factor, is implicated in various heart diseases, while ferroptosis (iron and lipid hydrogen peroxide-dependent regulated cell death) plays a crucial role in SCM progression. However, the specific crosstalk between CXCL4, ferroptosis, and SCM remains unelucidated. BALB/c mice were randomly divided into six groups (Control, LPS, LPS + Sodium Cromoglycate (CS), LPS + Ferrostatin-1 (Fer-1), LPS + Pifithrin-α (PFT-α), LPS + Niclosamide) to establish the SCM model via intraperitoneal LPS injection. In vivo experiments included histopathological examination (H&E, toluidine blue staining), survival analysis, ELISA, Western blot, immunofluorescence, immunohistochemistry, TUNEL staining, and detection of myocardial markers (CK-MB, AST, LDH) and oxidative stress indicators (SOD, MDA, iron content). In vitro, RAW264.7 macrophages were treated with CXCL4 alone or combined with inhibitors (Fer-1, PFT-α, Niclosamide), followed by CCK-8 assay, ROS detection, qRT-PCR, Western blot, and phagocytosis microbead assay. In vivo, SCM mice exhibited significantly elevated CXCL4 levels in serum and heart tissue, accompanied by mast cell activation and degranulation. Inhibiting mast cell activation (with CS) reduced CXCL4 production, alleviated cardiac inflammation and ferroptosis (increased SLC7A11/GPX4 expression, decreased 4-HNE), and improved survival. TUNEL staining revealed predominant macrophage death in SCM hearts. In vitro, CXCL4 induced macrophage ferroptosis (downregulated SLC7A11/GPX4) and impaired phagocytic function (reduced CD36/MERTK expression), which was reversed by Fer-1. Mechanistically, CXCL4 activated STAT3 phosphorylation, regulating downstream P53; inhibiting STAT3 (Niclosamide) or P53 (PFT-α) alleviated macrophage ferroptosis, restored phagocytosis, and mitigated cardiac injury in SCM mice. Mast cell-derived CXCL4 induces macrophage ferroptosis via the STAT3/P53 signaling pathway, impairs macrophage phagocytic function, and exacerbates myocardial injury in SCM. Targeting mast cell activation, CXCL4 release, or the STAT3/P53-ferroptosis axis may serve as promising therapeutic strategies for SCM. Not applicable.