Vascular occlusions are a major cause of cardiovascular disease and require interventions including graft bypass surgery; yet, the development of synthetic vascular grafts remains a challenge due to the inability of synthetic biomaterials to support an anti-immunogenic endothelium. Topographical micropatterning of biophysical cues offers a promising strategy to enhance graft endothelialization. As the interface between the biomaterial and blood, endothelial cells provide a bioactive interface which regulates inflammation. We utilized topographical micropatterning of polyurethane biomaterial to induce endothelial cell elongation and alignment and measured the resultant transcriptional changes using bulk RNA-sequencing. Cell phenotype was characterized by gene and protein expression as well as chemokine secretion. The biomaterial micropatterning-modulated elongated and aligned morphology promoted an anti-inflammatory transcriptome, reduced pro-inflammatory chemokine secretion, and induced the transcription coregulator yes-associated protein (YAP) phosphorylation, supporting the anti-immunogenic properties of endothelial cells. Cytoskeletal regulation of this anti-immunogenic phenotype was quantified through measurement of intermediate filament vimentin regulation of YAP localization and endothelial pro-immunogenic gene expression. Aggregation of vimentin was positively correlated with YAP dephosphorylation and suppressed proinflammatory gene expression, indicating that endothelial elongation and alignment influence gene regulation and cellular function independent of vimentin-mediated mechanisms. The ability of micropatterned endothelial cells to maintain their anti-immunogenic functions under pro-inflammatory conditions was demonstrated by the increased phosphorylation of YAP and decrease in pro-inflammatory IL-8 release. Our study highlights the potential of biomaterial cues to regulate endothelial cell behavior for vascular graft design. STATEMENT OF SIGNIFICANCE: We used topographical micropatterning to separate the role of endothelial monolayer morphology from unidirectional blood flow and test how architecture alone regulates gene programs. Micropatterned substrates produced a reproducibly elongated, aligned endothelium with coordinated modeling of the vimentin intermediate filament network. Bulk transcriptomics revealed a distinct morphology-associated expression state, including reduced inflammatory signaling and altered secretory profiles, alongside shifts in mechanosensitive regulatory programs and downstream transcriptional signatures. Together, these findings link cytoskeletal organization to endothelial mechanotransduction and highlight biomaterial topography as a design lever to tune endothelial function for preclinical and clinical applications.

Benzo[a]pyrene (B[a]P), a prominent persistent organic pollutant that is widespread in marine environment due to anthropogenic activities, poses considerable threats to marine ecosystem and public health. Although prior research has documented the adverse impacts of B[a]P on organisms and their response to relieve injury, there are relatively few studies on marine crustaceans that show commercial significance. In this study, we investigated the dose- and time-dependent effects of B[a]P on the lipid peroxidation and gene expression in the hepatopancreas and gill of Marsupenaeus japonicus. The results revealed that malondialdehyde contents in the hepatopancreas and gills of M. japonicus increased at 5 μg/L and 50 μg/L B[a]P exposure, indicating that higher B[a]P concentrations could induce lipid peroxidation. Exposure to 50 μg/L B[a]P resulted in the downregulated expression of several antimicrobial protein genes, including C-type lectins, i-type lysozyme, and stylicin. In contrast, B[a]P exposure significantly upregulated the gene expression of heat shock proteins and antioxidant enzymes. Moreover, correlation analysis suggested that malondialdehyde (MDA) content and gene expression levels in the hepatopancreas and gill of M. japonicus could serve as candidate biomarkers for B[a]P. Overall, this study provided basic insights into the effects of B[a]P on oxidative stress and gene expression in M. japonicus, enriched the understanding of molecular mechanisms in crustaceans responding to B[a]P stress, and identified candidate biomarkers for B[a]P.

Nanoplastics (NPs) and heat stress co-occur in agricultural systems, yet their interactive effects on crops under different heat regimes remain unclear. Here, we investigated rice (Oryza sativa L.) responses to polystyrene NPs under chronic (CH; 36°C, 10 d) and acute (AH; 45°C, 3 h) heat stress using integrated transcriptomics, metabolomics, and alternative splicing analyses. Combined stressors produced additive biomass reductions; however, molecular responses exhibited regime-dependent non-additivity. Under CH, NPs induced widespread antagonistic interactions, suppressing the transcriptional acclimation program by ∼50%. Rather than alleviating stress, this antagonism reflected a systemic failure of heat defense activation, mediated by the disruption of a core regulatory module governing cell wall biosynthesis, phenylpropanoid production, and oxidative stress responses. This resulted in depletion of structural phospholipids and defense compounds (e.g., sakuranetin). Under AH, NPs altered the alternative splicing of circadian clock genes (OsCRY1, OsPRR73, OsGI). These findings reveal that NPs induce regime-specific, non-additive molecular effects despite additive phenotypic responses, demonstrating that traditional organism-level endpoints (e.g., biomass) lack the sensitivity to detect substantial disruption of molecular acclimation programs. This work highlights the need for integrating quantitative interaction modeling and systems-level analyses into multi-stressor risk assessments for agricultural systems facing concurrent pollution and climate stress.

Cellular stresses regulate transcriptional readthrough, whereby RNA polymerase II elongates past a gene's polyadenylation cleavage site without RNA cleavage. Readthrough has been reported in several cancer types. Here, we use long-read sequencing of nascent RNA to quantify transcriptional readthrough in chronic myeloid leukemia (CML) cells and characterize early responses to the targeted therapeutic, imatinib. We show that the amount, length, and gene specificity of readthrough increase within 1 hour, before gene expression and alternative splicing alterations emerge. Notably, imatinib-dependent messenger RNA (mRNA) isoform changes involved "readthrough chimeras," in which exons from an upstream gene are alternatively spliced to exons in a downstream gene. Altered mRNA isoforms and chimera levels were detected in imatinib-resistant K562 cells as well as cells of patients with CML. Thus, imatinib can provoke a cascade of early changes to transcription and splicing fidelity that may lead to longer-term adjustments in gene expression, cancer cell differentiation, and the development of therapy resistance.

Preoperative systemic chemotherapy plays a crucial role in enhancing the outcomes of patients with locally advanced esophageal squamous cell carcinoma (ESCC). Andrographis (major bioactive diterpenoid lactone isolated from Andrographis paniculata; PubChem ID: 5318517), a safe and cost‑effective dietary compound, has demonstrated antitumor effects against various gastrointestinal adenocarcinomas. However, its impact on squamous cell carcinoma remains unclear. The present study explored the antitumor effects of Andrographis and its potential to augment the antitumor efficacy of 5‑fluorouracil (5‑FU). A series of in vitro experiments was conducted, including cell proliferation, colony formation and apoptosis assays, using the ESCC cell lines KYSE410 and TE1. Compared with the controls, Andrographis significantly inhibited cell proliferation (P<0.05), suppressed colony formation (P<0.05), induced apoptosis (P<0.05), and upregulated the expression of ferroptosis‑related genes and proteins, such as HMOX1 (P<0.01), GCLC (P<0.05) and GCLM (P<0.001). Notably, even at a sub‑IC50 dose of 5‑FU, its combination with Andrographis resulted in additive antitumor effects (P<0.05) and further upregulation of ferroptosis‑related gene expression, particularly HMOX1 (P<0.05), compared with either mono‑treatment. The findings of the present study indicate that Andrographis exerts antitumor effects and enhances the efficacy of 5‑FU in ESCC by activating both apoptosis and ferroptosis, suggesting its potential as an adjunctive therapy for ESCC to improve efficacy and reduce 5‑FU dosage and toxicity.

Exposure to waste anesthetic gases (WAGs) is an underestimated occupational hazard in veterinary operating rooms (VORs), where insufficient ventilation and the absence of scavenging systems remain common worldwide. Veterinarians occupationally exposed to WAGs have been poorly investigated to date. Addressing this critical gap, we present the first integrative study evaluating circulating microRNAs (miRNAs), global DNA methylation, and urinary anesthetic quantification in veterinarians occupationally exposed to WAGs isoflurane and sevoflurane. In a case-control design, plasma profiling revealed 11 dysregulated miRNAs in the exposed group (n = 29) compared to the control group (n = 28) based on nominal p-values (p < 0.05), as part of an exploratory screening approach, including seven upregulated miRNAs meeting a predefined fold-change criterion (FC≥1.5). Among these, hsa-miR-1252-5p and hsa-miR-520f-3p showed robust discriminatory performance based on Receiver Operating Characteristic (ROC) curve analysis (AUC≥0.70). Functional enrichment analysis highlighted epigenetic regulators as major network hubs. For hsa-miR-1252-5p, hubs included EP300, TP53, CREBBP, HDAC1 and SIRT1, linking miRNAs modulation to histone acetylation/deacetylation, DNA damage response, apoptosis, and stress regulation. For hsa-miR-520f-3p, included MAPK1, TNRC6B, MECP2 and KMT2A, associated with cell signaling pathways, proliferation and epigenetic regulation. Global DNA methylation levels did not differ significantly between groups, suggesting that exposure under the evaluated conditions may not trigger genome-wide alterations. Occupational exposure was confirmed by urinary quantification of isoflurane and sevoflurane, indicating highly polluted workplaces. In conclusion, although global DNA methylation remained unchanged, WAG exposure was associated with modulation of circulating miRNAs, and our findings suggest that hsa-miR-1252-5p and hsa-miR-520f-3p emerge as potential biomarkers of effect associated to WAG occupational exposure in veterinarians who work in inadequately equipped VORs.

Emodin, a natural compound derived mainly from Reynoutria japonica (Huzhang), has demonstrated notable antitumor effects, though its epigenetic regulatory mechanisms in cervical cancer remain largely unclear. In this study, we show that emodin suppresses cervical cancer cell proliferation and induces oxidative stress. Integrated transcriptomic and metabolomic analyses further demonstrated that emodin markedly disrupted lipid metabolism, particularly by increasing phospholipid accumulation and promoting phospholipid peroxidation. Epigenetic profiling using Western blotting showed that emodin treatment altered global histone modification patterns, with notable increases in H3K27ac and H3K27me3 levels. Subsequent CUT&Tag analysis combined with ChIP-qPCR validation indicated that emodin modulates these two histone marks to regulate the transcription of key phospholipid metabolism-related genes, thereby contributing to enhanced phospholipid peroxidation and antitumor activity. Together, these findings provide new insights into the epigenetic antitumor mechanisms of emodin and highlight potential histone modification targets involved in phospholipid metabolism.

Body axial curvature is a common toxic effect of fish after exposure to environmental pollutants, but the molecular mechanism remains unclear. To investigate the underlying mechanisms, zebrafish were exposed to different persistent organic pollutants (POPs) including polychlorinated biphenyl (PCB) and 2,2',4,4'-tetrabromodiphenyl ether (BDE-47). PCB40 was chosen as the typical POP to discover the mechanism because of its remarkable body axial developmental toxicity. The PCB40 exposure reduced the density of spinal cord cilia. We observed the upregulation of neuropeptides urotensin II-related peptide (urp2), which is downstream of cilia signaling. Further analysis revealed an upregulation of lysine (K)-specific demethylase 1a (Kdm1a, also known as LSD1), and the microscale thermophoresis and chromatin immunoprecipitation assays indicated Kdm1a directly bound PCB40 and enriched in urp2 promoter regions to enhance urp2 expression. Both interventions of Kdm1a using a specific inhibitor and CRISPR/Cas9-mediated knockout partially rescued the body axial curvature induced by PCB40. Further molecular docking suggested that the previously reported pollutants which induced zebrafish body axial curvature could interact with Kdm1a. We found that PCB95 and BDE-47 also induced body axial curvature through Kdm1a and urp2. In conclusion, the study identified a previously unrecognized molecular pathway, specific POPs induced zebrafish body axial curvature by binding to Kdm1a, with urp2 emerging as a key candidate downstream mediator via Kdm1a-dependent upregulation. The study provides a new mechanism for fish body axial curvature caused by specific POPs and provides new insights for pollutant toxicity evaluation and prevention.

The effectiveness of immunotherapy is significantly limited by the inherently low immunogenicity and immunosuppressive phenotypes of tumors. To address this challenge, we develop assemblies composed of chlorin e6 and regorafenib (designated as CeRe), which combine photodynamic therapy (PDT) with immune regulatiing functions. CeRe exhibits uniform nanoscale distribution and good stability without requiring additional carriers. Upon light activation, CeRe eradicate tumor cells via PDT-induced reactive oxygen species (ROS) while simultaneously triggering immunogenic cell death (ICD). Furthermore, CeRe downregulates PD-L1 expression in tumor cells, promotes macrophage polarization, and relieves the immunosuppressive tumor microenvironment (TME). These synergistic immunomodulatory effects substantially improve tumor responsiveness to αPD-L1 treatment, leading to the effective inhibition of both primary and metastatic tumor growth. Collectively, this work presents a carrier-free nanodrug assembly strategy with multifaceted mechanisms, offering a promising approach for precise tumor therapy and metastasis suppression.

Evidence suggests that environmental exposures induce epigenetic modifications that can have long-lasting effects on multiple health outcomes, and an in-depth review of the epidemiological evidence is urgent. We aimed to comprehensively assess the associations between long-term exposure to air pollution and epigenetic changes in adults.

Plant-specific HD2s have been characterized; HDT1 orthologs were dicot-specific. AtHDT4 interacts with AtILR3 to increase Cd tolerance. The plant-specific histone deacetylase 2 (HD2) family is crucial for growth and stress responses, yet its evolutionary origins and functional diversification remain largely unknown. Here, we systematically elucidated the evolutionary trajectories of HDT homologs. We found the emergence of HDT1 orthologs as a dicot-specific innovation, characterized by unique motif acquisition and accompanied by relaxed purifying selection. Synteny analysis indicated that the duplication events establishing the HDT1 and HDT3 lineages were associated with whole-genome duplications (WGDs) specific to the dicot lineage. In contrast, the expansion of the HDT gene family in monocots appears to rely primarily on local duplication mechanisms, such as tandem duplications. Codon usage analysis revealed distinct species-specific preferences: lycophytes, bryophytes, and algae exhibited higher frequencies of G3s, C3s, GC3, CBI, Nc, and overall GC content, suggesting potential adaptive evolution or optimization for translational efficiency. Functional validation demonstrated that AtHDT4 contributes to the plant response to cadmium (Cd) stress. Specifically, AtHDT4 expression was significantly upregulated under CdCl₂ treatment. Compared with wild-type (WT) plants, the hdt4 mutant exhibited markedly reduced activities of key antioxidant enzymes, including superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT). Inductively coupled plasma mass spectrometry (ICP-MS) analyses confirmed that AtHDT4 regulates Cd accumulation. Furthermore, AtILR3 expression was significantly downregulated in the hdt4 mutant, implicating it in the Cd stress response. As anticipated, direct protein-protein interaction between AtHDT4 and AtILR3 was verified. This study not only uncovers the critical role of AtHDT4 in mediating plant responses to Cd stress but also provides a broader evolutionary perspective on the functional diversification and specialization of HDT homologs across plant lineages.

SUMOylation regulates chromatin states and transcriptional programs that preserve cellular identity, yet how perturbation of the SUMOylation pathway impacts adipocyte plasticity remains unclear. Here, we show that brief pharmacologic inhibition of SUMOylation in human pre-adipocytes using TAK-981 primes stable de novo beige differentiation in the presence of the PPARG agonist rosiglitazone. Transient TAK-981 exposure produces changes in the transcriptome and metabolism of mature adipocytes, including robust induction of canonical beiging markers like UCP1 and increased mitochondrial respiration. Mechanistically, ATAC-seq and RNA-sequencing revealed immediate chromatin remodeling and early mobilization of CEBP family members, followed by stable activation of CEBPA and PPARG regulatory networks. ChIP experiments demonstrated loss of H3K27me3 and gain of H3K27ac at PPAR response elements at thermogenic enhancers, and increased PPARG occupancy across the UCP1 regulatory unit. This mechanism is enforced by enhanced cAMP-PKA-p38 signaling and stabilization of beiging transcription activators. We propose that transient relief of SUMO-mediated repression unlocks dominant regulatory units, notably the UCP1 enhancer cluster, producing a monomorphic reprogramming toward adaptive thermogenesis. These findings identify SUMOylation as a reversible epigenetic barrier to adipocyte beiging and suggest that temporally controlled SUMO pathway inhibition combined with PPARG activation could be exploited to modulate adipose tissue thermogenic capacity.

The oomycete Phytophthora capsici causes Phytophthora blight, a major constraint on global pepper production. Our previous observations indicated that pretreating plants with thiamethoxam (TMX) and imidacloprid (IMI) could reduce the incidence of pepper blight, but the underlying mechanisms remained unclear. Here, we investigated how TMX and IMI induced resistance in pepper (Capsicum frutescens) against P. capsici. Both in vitro and in vivo assays demonstrated that TMX and IMI suppressed disease, not by directly impairing pathogen virulence but by inducing systemic resistance in susceptible (Cusheng L09) and resistant (Cusheng 356) pepper cultivars. Split-plant systemic resistance assays showed that TMX/IMI-primed plants developed smaller lesions in both treated and untreated leaves following P. capsici infection. Foliar application of TMX and IMI effectively alleviated disease severity, with IMI showing superior efficacy in attenuating reactive oxygen species (ROS) accumulation, and TMX/IMI priming concomitantly altering the activities of ROS-scavenging enzymes under pathogen challenge. Reverse transcription-quantitative PCR analysis revealed time-dependent changes in defence gene expression, and whole-genome transcriptome profiling highlighted temporal reprogramming of pathogenesis-related genes. Further functional validation identified CaNEN4 as a susceptibility factor. Collectively, our findings reveal that IMI/TMX primes pepper plants with systemic resistance by modulating ROS homeostasis, defence gene expression, and susceptibility gene function, offering novel insights into chemical-induced plant immunity and genetic targets for durable blight resistance in crops.

EGFR, one of the most successful therapeutic targets, has recently been found to exert a novel function for regulating homologous recombination (HR). Activation of HR is the critical event of treatment failure of PARPi in BRCA1/2 wild-type ovarian cancer (OC). Besides, the antitumor effects of biguanides have also been a focus of attention. Here, we discovered that the new biguanide 4C inhibited HR and sensitized BRCA1/2 wild-type OC cells to PARPi by targeting EGFR. Mechanistically, EGFR promoted nuclear accumulation of both BRCA2 and Rad51, and HR activation by competitively inhibiting the binding of BRCA2 and Rad51 to E3 ubiquitin ligase c-Cbl, thereby reducing cancer cell sensitivity to PARPi following ATM-mediated DNA damage signal transmission from the nucleus to the cytoplasm. Interestingly, EGFR was downregulated by 4C, which in turn enhanced the interaction of BRCA2 and Rad51 with c-Cbl. Consequently, BRCA2 and Rad51 were then ubiquitinated and degraded to inhibit HR and increase the sensitivity of OC to PARPi. Thus, these findings reveal that the combination of 4C with PARPi leading to "synthetic lethality" is an effective strategy for treating BRCA1/2 wild-type OC.

Dihuang Yinzi (DY) is a classic formula traditionally used for stroke-related disorders. Its potential therapeutic effect on post-stroke depression (PSD), however, remains to be investigated.

The mouse double minute 2 homolog (MDM2) - p53 interaction inactivates p53, a key tumor suppressor, and has been validated as a target for cancer therapy. Four triazolyl-thio-oxazines 4(a-d) were rationally designed based on a three-finger pharmacophore model, which parallels the binding conformation of p53 to suppress this oncogenic interaction. All compounds were characterized by 1H/13C NMR, IR, and mass spectrometry, whereas 4a was characterized by X-ray crystallography. All compounds bound to the MDM2 binding site at key residues (Leu54, Leu57, Ile61, Met62, Phe86, and Tyr100) with similar binding affinities (-7.88 to -8.75 kcal/mol). Further 200 ns molecular dynamics simulations and MM-GBSA analyses were performed to determine the precise differences. Complex 4a demonstrated better binding stability (RMSD <2.0 Å) and MM-GBSA Binding Score ΔGbind (-172.718 kcal/mol) than either Nutlin-3a or the p53 peptide. Its fluorobenzene dives further into the MDM2 cleft than the chlorobenzene ring in Nutlin-3a, resulting in stronger π-π stacking and hydrophobic interactions. SwissADME profiling also predicted that 4a has a good balance of pharmacokinetic properties. This was evident from the cytotoxicity studies (MCF-7 breast cancer cells), which revealed 4a to be the most potent (IC₅₀ = 4.379 μM). Further Western blot analysis showed significant upregulation of p53 and p21 in MCF-7 cells upon treatment with 4(a-d). In contrast, no significant induction of p53 or p21 was observed in MDA-MB-468 cells, confirming the upregulation of wild-type p53. These results suggest that 4a is a promising candidate for further biological evaluation as an MDM2-p53 interaction inhibitor.

Children with favorable-histology Wilms tumor (FHWT) who relapse or whose tumors show blastemal predominance post-chemotherapy often face poor outcomes. The purpose of this study is to identify mechanisms of chemotherapy resistance in FHWT. We induce a patient-derived xenograft model (KT-47) to develop blastemal predominance after chemotherapy and to become resistant to vincristine, actinomycin-D, and doxorubicin (VAD). Multi-omics analyses reveal chromatin and transcriptional changes, including increased H3K4me3 and decreased H3K27me3 at stem cell and nephrogenesis gene loci. LIN28B is the most upregulated resistance-associated gene, linked to MYCN copy gain/upregulation and chromatin remodeling. ABCB1 expression correlates with interchromosomal enhancer interactions and functions as the mediator of chemotherapy resistance in vitro. These findings are validated in additional Wilms tumor models. Overall, resistance is associated with de-differentiation to a stem-like state and is driven by ABCB1 upregulation, suggesting that therapeutic strategies targeting chromatin regulation and drug efflux may be relevant in therapy-resistant Wilms tumor.

Eradicating leukemia stem cells (LSCs) and overcoming tyrosine kinase inhibitor (TKI) resistance is urgent for chronic myeloid leukemia (CML) treatment. We find that F-box protein 3 (FBXO3) is highly upregulated in CD34+ CML stem cells from TKI-resistant patients and identify it as an innovative CML-LSC marker via single-cell RNA sequencing (scRNA-seq). FBXO3 deficiency induces apoptosis and reduces proliferation of CML cell lines and LSCs in vitro and in vivo, with minimal effects on normal CD34+ hematopoietic stem cells (HSCs). Mechanistically, FBXO3 interacts with DUSP9 to promote its ubiquitination and activate the MAPK pathway, critical for CML cell activity. DUSP9 knockdown partially reverses FBXO3-deficiency-mediated LSC elimination. Furthermore, FBXO3 inhibitor monotherapy or combination with imatinib effectively eradicates CML-LSCs, overcomes TKI resistance, and spares normal hematopoiesis. Collectively, our findings highlight FBXO3's role in CML progression and support combining FBXO3 inhibitors with TKIs for durable LSC elimination.

The application of salt-tolerant plant growth-promoting rhizobacteria (PGPR) represents a promising strategy to alleviate salt stress in crops. However, the mechanisms by which volatile organic compounds (VOCs) from actinomycetes mitigate salinity stress remain unclear. In this study, exposure to VOCs from the halotolerant actinomycete Glutamicibacter halophytocola KLBMP 5180 significantly promotes the growth of tomato seedlings under salt stress, as evidenced by increased fresh weight, lateral root number, and chlorophyll content. Physiological analyses show that VOCs exposure reduces oxidative damage and enhances antioxidant enzyme activity. Furthermore, elevated levels of total phenolics and flavonoids are detected, along with decreased sodium ion accumulation and an improved Na⁺/K⁺ ratio. Transcriptome analysis reveales that VOCs treatment upregulates genes associated with pathways involved in phenylpropanoid biosynthesis, glutathione metabolism, MAPK signaling, plant hormone signal transduction, and plant-pathogen interactions under saline conditions. VOCs also increase levels of endogenous auxin, jasmonic acid (JA), and 1-aminocyclopropane-1-carboxylic acid (ACC), while activating genes related to their respective signaling pathways. The salt tolerance enhancement mediated by VOCs is compromised by ethylene and JA inhibitors, suggesting that KLBMP 5180-derived VOCs improve tomato salt tolerance by stimulating ethylene synthesis and signal transduction. Additional VOCs profiling identifies several key bioactive compounds, including 3-methyl-1-butanol, phenylethyl alcohol, 2-aminopropanediamide, and 2-undecanone. These findings demonstrate that VOCs from G. halophytocola KLBMP 5180 enhance tomato growth under salt stress and advance our understanding of the role of actinomycetes in pronoting plant growth and mitigating salinity stress.

Hydrogen sulfide (H2S) has been recognized for its potent neuroprotective effects in Parkinson's disease (PD) models, yet the underlying cellular and molecular mechanisms remain to be fully elucidated. Ferroptosis, an iron-dependent form of regulated cell death, is increasingly implicated in the pathogenesis of PD. Isthmin 1 (ISM1) is a secreted protein predominantly expressed in the brain with emerging roles in neuroprotection and metabolic regulation. This study aimed to investigate whether H2S protects against rotenone (ROT)-induced neuronal injury by modulating ferroptosis and to explore the potential mediating role of ISM1 in this process.