Rotenone (RTN), an insecticide that functions through inhibiting mitochondrial electron transport chain complex I, has been demonstrated to impair reproductive health in mammals. Nevertheless, its impact on the process of preimplantation development remains to be elucidated. In this study, the toxic effects of RTN on mouse preimplantation embryo development were determined by exposure to different concentrations of RTN. RTN exposure resulted in developmental arrest of mouse preimplantation embryos in a dose-dependent manner. Mechanistically, RTN treatment induced mitochondrial dysfunction, elevated ROS levels, severe DNA damage and developmental arrest at the 2-cell stage. Furthermore, RTN treatment resulted in the failure of nuclear localization of the mitochondrial TCA cycle enzyme, pyruvate dehydrogenase (PDH), and abnormalities in histone modifications. This, in turn, led to impaired zygotic genome activation and development beyond the 2-cell stage. In addition, dichloroacetic acid enhanced the nuclear localization of PDH in 2-cell embryo, thereby augmenting TCA flux and promoting preimplantation development. In conclusion, RTN has the potential to exert cytotoxic effects on mammalian reproduction, and electron transport chain complex I inhibitors should be used with caution when applied to the treatment of reproductive diseases. Furthermore, this study enhances our comprehension of the interplay between mitochondrial metabolism and epigenetic regulation in early embryos.

HIPK2 is a nuclear serine/threonine kinase that regulates development and cellular stress responses including DNA damage response. Previously we have shown that HDAC6 increases HIPK2 protein stability via UBP domain rather than its deacetylase activity. Here, we identify USP2 as a deubiquitinase that stabilizes HIPK2 by complexing to HDAC6 UBP. Overexpression of USP2 increased while its knockdown or chemical inhibition decreased the protein level of HIPK2. USP2 decreased the polyubiquitination of HIPK2, suggesting that USP2 is a HIPK2 deubiquitinase. We found evidence that HDAC6, USP2 and HIPK2 form complex in cells. Importantly, USP2 knockdown or chemical inhibition suppressed the HDAC6-mediated accumulation of HIPK2 protein. This indicates that the HDAC6-mediated HIPK2 regulation is USP2-dependent. Functionally, upregulation of HIPK2 by USP2 promoted the TNFα-induced cell death. Overall, our data suggest that USP2 increases HIPK2 protein stability and enhances its proapoptotic function.

The mechanism of COVID-19-associated acute kidney injury (AKI) remains incompletely understood. This study aimed to investigate the regulatory role of the C5a-C5aR1 axis in neutrophil extracellular trap (NET) formation in SARS-CoV-2 nucleocapsid (N) protein-induced AKI. An AKI mouse model with kidney-specific N protein expression was established using ultrasound-microbubble-mediated technology, and interventions with the C5aR1 inhibitor DF2593A or DNase I (Pulmozyme) were administered to detect renal neutrophil infiltration, NET deposition, C5a-C5aR1 expression, and renal function indices. The results showed that N protein expression significantly induced neutrophil infiltration and NET formation, accompanied by elevated serum C5a and upregulated C5aR1. Inhibition of C5aR1 or degradation of NETs effectively attenuated renal tubular injury and decreased serum creatinine, blood urea nitrogen, and kidney injury molecule-1 (KIM-1) levels. In conclusion, activation of the C5a-C5aR1 axis contributes to N protein-mediated AKI by driving NET formation, and targeting this axis may provide a potential therapeutic strategy for COVID-19-associated renal injury.

Hypoxia promotes ovarian cancer progression, EMT and platinum resistance, but the lncRNA mediators involved remain unclear. Here, we identified ZNF213-AS1 as a downregulated lncRNA in ovarian cancer by analyzing TCGA-OV and GTEx datasets. Low ZNF213-AS1 expression predicted poor overall survival and was associated with hypoxia, EMT, NF-κB and IL6/JAK/STAT3 signatures. ZNF213-AS1 was relatively abundant and enriched in the chromatin-associated fraction. Hypoxia reduced ZNF213-AS1 expression in ovarian cancer cells, but this repression was not rescued by HIF1α knockout or mutation of the predicted HRE motif, suggesting a noncanonical hypoxic regulatory mechanism. Functionally, ZNF213-AS1 overexpression suppressed hypoxia-induced colony formation, invasion and cisplatin resistance, whereas ZNF213-AS1 knockdown enhanced these phenotypes. Mechanistically, ZNF213-AS1 inhibited NF-κB reporter activity and reduced NF-κB/STAT3/EMT-related gene expression. RNA pull-down and RIP assays showed that ZNF213-AS1 associates with RELA/p65-containing NF-κB complexes. Moreover, p65 re-expression partially reversed the inhibitory effects of ZNF213-AS1 on STAT3 activation, EMT markers and xenograft growth. These findings define a hypoxia-repressed ZNF213-AS1-p65 axis that limits NF-κB/STAT3-driven EMT and platinum-resistant ovarian cancer progression.

Environmental salinity instability associated with climate change increasingly challenges aquatic organisms inhabiting marine and estuarine ecosystems. Although osmotic acclimation mechanisms in euryhaline fish have been extensively investigated, whether ancestral osmotic stress exposure alters offspring osmotic responsiveness remains poorly understood. In the present study, marine medaka (Oryzias melastigma) derived from osmotic stress-experienced (EXP) and non-experienced (Non-Exp) F2 lineages were subjected to acute hypotonic challenge before gill RNA-sequencing analysis. Acute hypotonic exposure induced broader transcriptomic remodeling in the Non-Exp lineage, whereas the Exp lineage exhibited a smaller but more condensed enrichment profile associated with osmosensing- and ion transport-related pathways. The Non-Exp lineage predominantly enriched pathways related to RNA transcription, membrane trafficking, epithelial remodeling, and stress-associated signaling. In contrast, the Exp lineage preferentially enriched pathways associated with TRP channels, stimuli-sensing channels, ion channel transport, and SLC-mediated transmembrane transport. Representative osmoregulatory transcripts including trpv4, aqp3, and slc12a1 were additionally altered in the Exp lineage. Collectively, these findings suggest that ancestral osmotic stress exposure may influence offspring transcriptomic responses to acute hypotonic challenge and provide evidence for transgenerational plasticity in osmotic acclimation.

Nanoplastics have emerged as important environmental contaminants with potential risks to animal and human health; however, their effects on intestinal epithelial cells remain unclear. In this study, polystyrene nanoplastics (PS) were used to investigate cytotoxic mechanisms in porcine intestinal epithelial cells(IPI-2I). PS exposure was associated with oxidative stress, as evidenced by elevated reactive oxygen species levels, altered oxidative stress-related indicators, and mitochondrial structural alterations. Increased γ-H2AX expression suggested activation of DNA damage responses. Transcriptomic analysis revealed enrichment of pathways associated with oxidative stress and telomere maintenance. PS exposure was also associated with telomere-associated alterations, including altered expression of shelterin complex proteins (TERF1, TERF2, and POT1), decreased telomerase activity, and shortened relative telomere length. In addition, PS treatment was associated with senescence-related phenotypes, including increased p53 and p21 expression and a higher proportion of SA-β-gal-positive cells. Bioinformatic and experimental analyses suggested a potential association between PPARα and POT1 regulation under PS exposure conditions. Chromatin immunoprecipitation-qPCR analysis suggested that PPARα could associate with the POT1 promoter region, and this enrichment was reduced after PS exposure. Collectively, the present findings suggest that PS exposure is associated with oxidative stress, DNA damage, telomere-associated alterations, and senescence-related changes in intestinal epithelial cells. In addition, PPARα may participate in telomere-associated cellular responses under PS exposure conditions.

Hypercholesterolemia (HCh) represents a prevalent comorbidity in patients with diabetic foot ulcers (DFUs), frequently exacerbating wound recalcitrance, yet the mechanisms linking systemic lipid perturbation to local immune dysregulation remain poorly defined. Here, we applied an integrated transcriptomic and machine learning approach to explore the molecular basis of this comorbidity. Initial analyses identified 70 shared DEGs between DFUs and HCh, primarily enriched in pathways related to lipid homeostasis disruption and inflammatory cascade activation. Subsequently, a four-algorithm machine learning pipeline highlighted AKR1C3 and CLIC3 as candidate diagnostic biomarkers, both consistently downregulated across disease states and supported by independent external validation cohorts (AUC = 0.779 and 0.920). Immune deconvolution further indicated a dysregulated microenvironment featuring sustained innate immune activation alongside adaptive immune impairment, which correlated with reduced AKR1C3 and CLIC3 expression. Drug repurposing analyses proposed that pirfenidone and statins might attenuate this pathological transcriptomic signature, while molecular docking simulations suggested stable binding of exemestane and indomethacin to AKR1C3 (binding energies: -8.0 and -7.3 kcal/mol, respectively). Collectively, these findings suggest that AKR1C3 and CLIC3 act as molecular bridges linking the TGF-β/lipid metabolism axis to local immune dysfunction, offering a rationale for early risk stratification and targeted drug repurposing in DFUs-HCh comorbidity.

Mutations in the cardiac splicing factor RNA binding motif protein 20 (RBM20) are associated with severe forms of dilated cardiomyopathy. Mutations predominate within the highly conserved RSRSP-stretch and are characterized by a cytoplasmic mislocalization of RBM20. However, a sequence downstream of the RSRSP-stretch, the SRSLSP-stretch, is also phylogenetically highly conserved. The impact of the SRSLSP-stretch is unclear. In our study we investigated subcellular localization of RBM20 variants in HEK293 cells after transient transfection. We showed that variants within the RSRSP-stretch lead to significant cytoplasmic mislocalization. Aberrant TTN-splicing was observed in the explanted myocardium of an RBM20-p.R634W carrier. Within the SRSLSP-stretch, variants at amino acid position 641 are associated with partial mislocalization of RBM20. Molecular modelling of RBM20 and transportin-3 interaction by AlphaFold-3 suggests that the RSRSP stretch and arginine at position 641 are involved in transportin-3 interaction. In summary, our study supports the functional relevance of the RSRSP-stretch for the RBM20-TNPO3 interaction, whereas variants within the adjacent SRSLSP stretch have a minor impact on the nuclear localization. Furthermore, the data underscore the limitations of in silico-based pathogenicity predictions of RBM20 variants.

Associationof cationic liposomes to cells is not governed by their mean surface charge alone. Using flow cytometry and confocal microscopy, we show that coating DOTAP/DOPC liposomes with anionic carboxymethyl-β-cyclodextrin (β-CMCD) increases cell association: the fraction of liposome-associated cells rises from 35% to 45% for MSC and from 59% to 65% for MDA-MB-231 cells, with a shift from peripheral to more volume-coupled accumulation in cancer cells. Based on an energy-based model, we attribute this enhancement to the formation of a rough, patchy, multivalent β-CMCD shell. Despite partial screening of the average positive charge, local contact amplification by protrusions, charge-patch matching, cooperative surface accumulation and secondary cyclodextrin-lipid contacts outweigh the electrostatic loss. This resolves the apparent paradox: β-CMCD-coated liposomes show greater association with model membranes and cells than bare cationic liposomes despite their lower electrophoretic mobility.

3-Hydroxy-3-methylglutaryl-CoA reductase (HMGR) catalyzes the committed step of the mevalonate pathway and is encoded by multigene families in plants. However, the functional specialization of individual isoforms in abiotic stress remains poorly understood in legumes. Here, we characterize three HMGR isoforms from chickpea (Cicer arietinum) and demonstrate that CaHMGR2 and CaHMGR3 function as stress-inducible isoforms with structural and regulatory divergence from the constitutively expressed CaHMGR1. Quantitative expression profiling revealed strong induction of CaHMGR2 and CaHMGR3 under drought, salinity, and cold stress. Promoter analysis identified enrichment of abiotic stress-responsive cis-elements exclusively in these isoforms. Structural comparison uncovered conserved substitutions within the dimerization motif, transmembrane regions, and residues proximal to the regulatory serine, suggesting altered regulatory properties. Subcellular localization further distinguished CaHMGR2 and CaHMGR3, which associate with both endoplasmic reticulum and nuclear envelope. Functional validation using overexpression lines demonstrated enhanced germination, root growth, reduced water loss, and improved recovery under stress. Metabolic profiling and KEGG pathway enrichment revealed selective metabolic re-programming associated with branched chain amino acid and fatty acid metabolism. Together, these findings establish functional specialization of HMGR isoforms and their role in abiotic stress resilience in chickpea.

The regenerative capacity of the liver critically determines recovery outcomes following partial liver transplantation or hepatectomy. While cytokines, immune responses, and metabolic dynamics modulate hepatic regeneration, the role of aconitate decarboxylase 1 (Acod1)-a key enzyme catalyzing itaconate biosynthesis-remains underexplored. This study elucidates the regulatory function of Acod1 in liver regeneration and its underlying mechanisms. Male wild-type (WT) and Acod1-knockout (Acod1-/-) mice underwent 70% or 90% partial hepatectomy (PHx), with interventions including 4-octyl itaconate (4OI) and citraconate administration. Postoperative assessments at 0, 3, 6, 12, 24, 36 and 48 h included liver-to-body weight ratios, serum ALT/AST levels, histopathology, proliferation markers (Ki67, PCNA), cell cycle gene expression, and survival analysis. RNA seq and metabolic profiling were performed to explore mechanistic pathways. Our results demonstrated that Acod1 expression peaked at 36 h post-hepatectomy. In 90% lethal PHx model, Acod1-/- mice showed significantly improved survival versus WT controls. Following 70% PHx, Acod1-/- mice exhibited enhanced liver regeneration at 36 h, with significantly lower serum transaminase levels compared to WT controls. Exogenous 4-OI administration abrogated this pro-regenerative phenotype in Acod1-/- mice, whereas CITRA treatment in WT mice produced a similar pro-regenerative pattern, improving both regeneration-associated readouts and survival. Mechanistically, RNA-seq revealed upregulated ketone body metabolism genes in Acod1-/- mice, concomitant with elevated hepatic β-hydroxybutyrate and reduced intrahepatic lipids versus WT controls. This study demonstrates that Acod1 deficiency enhances post-hepatectomy liver regeneration by suppressing itaconate production, thereby promoting fatty acid mobilization. These findings support Acod1-associated metabolic regulation as a potential therapeutic target for optimizing liver regeneration and postoperative recovery.