Parotid gland aging is a key factor for reduced salivary production in older individuals. The role of 4-Octyl itaconate (4-OI) in age-related parotid gland dysfunction remains unclear. Thirteen-month-old male mice were administered 4-OI (25 mg/kg) or vehicle once daily for 4 weeks. The salivary flow rate was calculated to assess gland secretion function. Hematoxylin and eosin staining was used to analyze morphological alterations. Masson staining was used to quantify collagen deposition. Oil Red O staining was used to evaluate lipid droplet accumulation. Western blotting was used to detect protein expression levels. Metabolomics, transcriptome, and molecular docking analyses were performed to identify potential mechanisms of action. Functional assays revealed that 4-OI increased the salivary flow rate. AMPKα has been identified as a direct target of 4-OI by molecular docking. 4-OI upregulates AMPKα phosphorylation, decreases lipid droplet deposition, and relieves fibrosis to restore the aging characteristics of the gland. 4-OI regulates fatty acid oxidation (FAO) in the parotid gland through AMPKα/PPARα signaling pathways. The protective effects of 4-OI in aged mice were abolished by Compound C and GW6471, which inhibit AMPKα and PPARα, respectively. In addition, we found that 4-OI treatment ameliorated parotid gland aging-induced oxidative reactions, enhanced mitochondrial biogenesis, and activated the Nrf2 pathway. Moreover, the Nrf2 inhibitor ML385 was used to validate this mechanism. Treatment with 4-OI improved the aging of the parotid gland by increasing gland FAO, decreasing gland fibrosis, and preventing oxidative reactions. This was achieved by upregulating the AMPKα/Nrf2 and AMPKα/PPARα signaling pathways.

CD8[+] T cells play an important role in anti-infection and anti-tumor immunity. Metabolic reprogramming has emerged as a critical regulator of T cell function, yet the roles of specific cholesterol-associated proteins during T cell fate trajectories remain unclear. Here, using an in vivo CRISPR screen in CD8[+] T cells during infection with a customized sgRNA library targeting key cholesterol metabolism-associated proteins, we successfully identified several candidate targets, including the LBR (Lamin B Receptor). Lbr knockout significantly reduced the proportion and absolute number of short-lived effector cells (SLECs), severely impairing effector responses, while concomitantly increasing the memory precursor population upon LCMV infection. Furthermore, transcriptomic analysis revealed that Lbr depletion markedly altered signaling pathways governing effector T cell differentiation. These findings reveal the role of LBR in modulating effector and memory T cell differentiation, suggesting it as a potential target for metabolic reprogramming to enhance T cell-based immunotherapies.

As the leading cause of uterine factor infertility, intrauterine adhesion (IUA) poses a severe threat to the fertility and mental health of women at childbearing age. However, the efficacy of the present clinical treatments is unsatisfactory. There remains an urgent need to develop a safe, effective, and clinically feasible therapeutic strategy to restore the damaged endometrium. Here, we fabricated a gelatin-based electrospun nanofibrous membrane (GENM) via electrospinning and chemical crosslinking technique and characterized its properties. The anti-fibrotic and pro-angiogenic effects of GENM were evaluated in cellular models. A rat IUA model induced by mechanical scraping was established to assess the therapeutic efficacy of GENM by histological staining. Additionally, bulk RNA sequencing was performed to explore the underlying mechanisms. Our results demonstrated GENM inhibited transforming growth factor beta 1 (TGF-β1) induced fibrosis in human endometrial stromal cells (HESCs) and promoted angiogenesis of human umbilical vein endothelial cells (HUVECs). Animal experiments also suggested that GENM significantly improved endometrial receptivity and neovascularization while reducing fibrosis in the rat IUA model. RNA sequencing further identified several signaling pathways contributing to uncovering its mechanism of action. These findings indicate that GENM holds significant potential for clinical application in the management of IUA.

Well-regulated trophoblast proliferation, migration, invasion, and cell turnover are essential for normal placental development in humans and rodents. Given the established role of oxidative stress in placentation and the redox activity of iron, we investigated whether ferroptosis-associated redox signaling contributes to trophoblast function and placental development. Placental iron profiling revealed significantly elevated total and ferrous iron levels in first-trimester human villi compared with term placentas, accompanied by the transcript levels of several iron uptake- and reduction-related genes, including TFRC, DMT1, ZIP8, STEAP3, and STEAP4, which were elevated in first-trimester villi. Murine placentas also exhibited gestational changes in iron abundance and iron-homeostasis-related gene expression. HO-1 protein abundance was highest during early gestation and declined thereafter, suggesting a potential association between heme degradation and gestational iron homeostasis. In contrast, placental labile iron pool (LIP) levels showed only modest, statistically nonsignificant changes across gestation, suggesting a relatively stable redox-active iron pool. Immunofluorescence analyses demonstrated spatially distinct expression of ferroptosis-associated regulators, with ACSL4 enriched in invasive trophoblast populations and GPX4 predominantly localized in surrounding decidual tissues, suggesting regional heterogeneity in ferroptosis-associated molecular features during placentation. Functional studies in HTR-8/SVneo trophoblast cells further demonstrated that mild ferroptosis-associated redox perturbation induced by low-dose erastin or ferrous iron enhanced trophoblast migration and invasion without overt cytotoxicity. These effects were attenuated by ferrostatin-1, deferoxamine mesylate (DFOM), or the mitochondria-targeted antioxidant MitoQ and were blunted following FTH1 and TFRC knockdown, indicating that trophoblast responsiveness depends on iron availability and ferroptosis-associated redox signaling. Importantly, these pro-invasive effects were preserved under physiologically relevant hypoxic conditions. Together, our findings support a model in which gestational changes in placental iron homeostasis are associated with ferroptosis-related molecular features, while experimentally induced iron-dependent redox signaling modulates trophoblast behavior. Rather than inducing overt ferroptotic cell death, sublethal iron-dependent redox perturbation may act as a signaling mechanism influencing trophoblast function during placental development.

Preeclampsia (PE) affects 3%-5% of pregnancies worldwide and is a leading cause of maternal/perinatal mortality, yet its etiology remains unknown. We established a PE rat model using L-NAME to investigate postpartum cognitive dysfunction and its mechanisms, evaluated brain microstructural changes with MRI, explored correlations between MRI parameters and cognitive impairment, and assessed pravastatin's therapeutic effects. Sixty-three pregnant rats were divided into normal pregnant (NP), PE, and pravastatin-treated (PRA) groups, with three postpartum subgroups (10 days, 1 month, 3 months; n = 7 each). Morris water maze, HE/Nissl staining, transmission electron microscopy, and multimodal MRI (DTI, IVIM, DCE-MRI) were performed. ROIs included anterior/posterior cortex and hippocampus. Pearson correlation was used to relate MRI parameters to escape latency. (1) L-NAME induced a PE-like phenotype, alleviated by pravastatin. (2) Cognitive impairment appeared at 10 days postpartum and worsened over time. (3) FA, D, and f values decreased while Ktrans increased in the PE group (p < 0.05), changes that progressed over time and were abrogated by pravastatin. (4) FA and D were negatively correlated with escape latency, whereas Ktrans showed a positive correlation. (5) Neuronal degeneration, astrocyte activation, and BBB disruption in the PE group were ameliorated by pravastatin. PE model rats developed progressive postpartum cognitive dysfunction, likely due to neuronal degeneration, glial activation, and increased BBB permeability. Pravastatin partially alleviated these pathological and cognitive changes. Multimodal MRI effectively detected alterations in brain microstructure, microvascular perfusion, and BBB, consistent with histopathological findings.

Succinate is a key metabolic intermediate in the tricarboxylic acid (TCA) cycle, which participates in energy metabolism. It accumulates under various pathological conditions and exerts regulatory effects through binding to its specific receptor GPR91. However, its specific function in sepsis-associated acute kidney injury (SA-AKI) remains unknown. Herein, we utilized GPR91 knockout (GPR91-/-) mice and LPS-induced SA-AKI mouse models to elucidate the role and mechanisms of the succinate-GPR91 axis in SA-AKI. The results demonstrated significantly elevated succinate levels in the circulation of both SA-AKI patients and mice. Exogenous succinate supplementation markedly mitigated renal inflammatory responses and histopathological damage in SA-AKI mice. Notably, the deletion of GPR91 not only markedly exacerbated renal inflammation and tissue injury but also completely abolished the protective effects of exogenous succinate. Further mechanistic studies revealed that succinate suppresses activation of the PI3K-AKT signaling pathway in a GPR91-dependent manner, thereby reducing inflammation and attenuating renal damage in SA-AKI. In conclusion, this study is the first to demonstrate that the succinate-GPR91 axis exerts anti-inflammatory and renal protective effects by inhibiting the PI3K-AKT pathway, highlighting it as a potential therapeutic target for SA-AKI.

Integrating recent biological and epidemiological insights, this analysis re-examines the Linear No-Threshold (LNT) model-the current basis for radiological protection-which assumes that cancer risk increases proportionally with any dose of ionizing radiation (IR). Empirical data from a cross-sectional study of 5156 radiation workers indicated a lower frequency of micronuclei in lymphocytes at very low cumulative doses (< 1 mSv) compared to a pre-employment control group. This observation is suggestive of, but does not directly prove, an adaptive cellular response; importantly, micronuclei are downstream cytogenetic endpoints that do not directly measure DNA repair capacity or carcinogenic risk, and thus this finding cannot, by itself, be taken as evidence for enhanced repair or reduced cancer risk. Meanwhile, ecological studies in high-background radiation areas (Yangjiang, Kerala, Ramsar) have reported nonlinear dose-response patterns, although these ecological studies are subject to confounding and should be interpreted with caution. These findings raise the hypothesis that chronic, near-background exposures (~2-3 mSv/year) could trigger adaptive responses (radiation hormesis), a hypothesis that remains to be rigorously tested. Plausible mechanisms, which are speculative at this stage, include upregulated DNA repair (e.g., PARP1) and antioxidant systems (e.g., SOD), potentially enhancing radical scavenging and epigenetic adaptation, which could reduce genomic instability. However, the existence of a universal "practical threshold" cannot be concluded from the present observational data alone, and further mechanistic and prospective studies are needed. Therefore, it is possible that the low-dose regime follows a "practical threshold" or a J-shaped curve rather than a strictly linear relationship, although this remains an open scientific question. Future efforts should link the processing of complex damage to organism-level effects, including genomic instability and immune responses, in order to advance biologically grounded risk models.

Acute myeloid leukemia (AML) is a heterogeneous disease with large spectrum of specific mutations and gene aberrations. Recently, the Bcl-2 inhibitor Venetoclax, in combination with hypomethylating agents (HMAs), was approved for older (> 65 years) AML patients, as well as for those unfit for intensive induction chemotherapy. In addition to Bcl-2 inhibition, Venetoclax also induces generation of reactive oxygen species (ROS). We demonstrated that distinct fraction exhibiting specific features arises during 24 h of sample exposure to Venetoclax. This fraction displays characteristic preapoptotic markers as mitochondria depolarization and partial Annexin V surface positivity. Moreover, monitoring of ROS showed negative correlation between signals detected using H2DCFDA and CellROX probes pointing to dynamic ROS changes induced by Venetoclax. The addition of HMA (Decitabine) had almost no effect on cell viability or ROS production but caused proliferation arrest in sensitive cells. In our panel of AML cell lines and primary AML samples we have found a correlation between ROS production, markers of apoptosis, and attenuation of Bcl-2 activity after Venetoclax treatment. Level of Mcl-1, another antiapoptotic protein from the Bcl-2 family, was reduced in sensitive cells, but increased in the resistant samples in response to Venetoclax. Moreover, the nucleolar protein nucleolin (NCL), which is frequently overexpressed in AML cells, was significantly deregulated in Venetoclax-treated cells. In particular, both NCL protein level and specific phosphorylation decreased in fractions sensitive to Venetoclax. Our findings suggest that Venetoclax targets distinct cell subpopulation, and that ability of a cell to follow increased ROS drives its response to Venetoclax.

Cardiac aging is a major risk factor for cardiovascular diseases, with an imbalance in mitochondrial dynamics homeostasis being a pivotal aspect. Although Astragaloside IV (AS-IV) has been shown to delay aging, its specific effects on cardiac aging and the link to mitochondrial dynamics remain unclear. This study aimed to elucidate the role and regulatory mechanism of AS-IV in delaying cardiac aging by using naturally aged mouse models and D-galactose (D-gal)-induced H9c2 cell senescence models. The results demonstrated that AS-IV reduced the expression of senescence markers (p16[INK4a], p21, and p53), β-galactosidase (SA-β-gal), and senescence-associated secretory phenotype (SASP) factors (Il6, Il1b, and Tnfa) while also significantly ameliorating aging-induced cardiac dysfunction and myocardial fibrosis. Furthermore, AS-IV significantly enhanced mitochondrial morphology and function in cardiomyocytes. This improvement was demonstrated by increased mitochondrial membrane potential, elevated ATP production, and reduced ROS levels. Mechanistically, AS-IV upregulated SIRT3 and promoted its interaction with DRP1, leading to the inhibition of DRP1 acetylation. This action suppressed excessive mitochondrial fission and thereby improved mitochondrial dynamics. Transfection with SIRT3-siRNA attenuated the beneficial effects of AS-IV. In conclusion, this study demonstrates that AS-IV mitigates cardiac aging by activating SIRT3 to mediate DRP1 deacetylation, which in turn improves mitochondrial dynamics. These findings provide a strong theoretical and experimental foundation for the potential application of AS-IV in preventing and treating age-related cardiovascular diseases.

Mitochondrial apoptosis is a key regulator of tumor progression, therapeutic resistance, and immune evasion in non-small cell lung cancer (NSCLC), yet promising candidate biomarkers remain inadequately defined. Using an integrative multi-omics systems biology framework, we identified 16 mitochondrial apoptosis-associated genes and uncovered a highly interconnected apoptotic network centered on MCL1, BCL2L1, BAX, CASP8, and CASP9. Comprehensive transcriptomic, genomic, and epigenetic analyses revealed recurrent genomic alterations, subtype-specific dysregulation, copy number variation-driven transcriptional changes, differential DNA methylation, and significant survival associations, highlighting their clinical relevance. Independent validation across external transcriptomic cohorts confirmed robust differential expression of the prioritized biomarkers, while physiological transcriptomic and protein atlas analyses demonstrated conserved transcript architecture, widespread tissue expression, and mitochondrial localization of MCL1. Importantly, spatial transcriptomic profiling in a mouse NSCLC model experimentally provided complementary evidence consistent with the computational findings by demonstrating markedly increased and spatially heterogeneous expression of MCL1, BCL2L1, and CASP9 within tumor tissues and their enrichment in distinct cellular populations, supporting their role in tumor heterogeneity and mitochondrial apoptosis regulation. Integrative analysis consistently prioritized MCL1 as the dominant mitochondrial apoptosis biomarker, with BCL2L1 and CASP9 emerging as additional high-confidence candidates and CASP8 and BAX serving as complementary prognostic biomarkers. These findings establish MCL1 as a promising candidate biomarker and promising precision therapeutic target, while demonstrating that the integration of multi-omics discovery with spatial transcriptomic validation provides a comprehensive molecular framework for apoptosis-driven precision oncology in NSCLC.

A novel gold nanoparticle-chitosan (AuNP@chitosan) composite is introduced as a potent biointerface modulator for burn trauma care. In this study, the unique interfacial characteristics of the composite are linked to its therapeutic activity, and its effects on endothelial function and immune response in severe bleeding and inflammatory models are examined. AuNP@chitosan synthesis was verified using FTIR and fluorescence, and its effects were assessed in TNF-α-induced HUVECs, LPS-stimulated macrophages, and an animal model. In vitro, AuNP@chitosan protects TNF-α-injured human endothelial cells (HUVECs) by restoring thrombomodulin expression, reducing ROS, and relieving the upregulated hypoxia-adaptive markers (HIF-1α and fibronectin). Concurrently, in LPS-stimulated RAW 264.7 macrophages, it suppresses pro-inflammatory mediators (TNF-α and IL-6) while preserving COX-1 and enhancing IL-10 production. These cell-specific responses reflect distinct but complementary mechanisms, vascular protection and immune regulation, which were further validated in a rat burn model. In vivo application of AuNP@chitosan dressings significantly reduced infiltration of CD4[+], CD8[+], and CD68[+] immune cells in wound tissue, supporting the translational relevance of the dual in vitro findings. The integrated cell and animal studies demonstrate that AuNP@chitosan acts via redox and immune modulation to promote a pro-healing wound environment.

Female reproductive disorders are closely associated with ovarian oxidative stress and lipid dysregulation, but the upstream triggers remain poorly defined. In this study, we found that Cyp27b1, a key mitochondrial enzyme that catalyzes the conversion of 25-hydroxyvitamin D3 (25(OH)D3) to 1,25-dihydroxyvitamin D3 (1,25(OH)2D3), is an important regulator of ovarian lipid homeostasis and redox balance. In the cyp27b1[-/-] zebrafish model, cyp27b1 deletion impaired ovarian development and caused mitochondrial dysfunction and oxidative stress, all of which were alleviated by exogenous 1,25(OH)2D3 treatment. Lipidomic analysis revealed an abnormal accumulation of triglycerides (TG) and diglycerides (DG) in the knockout group, while phosphatidylcholine (PC) and phosphatidylethanolamine (PE), key membrane lipid components, were significantly reduced, disrupting membrane integrity and energy homeostasis. Dual-luciferase reporter assays showed that 1,25(OH)2D3 increased pnpla2 promoter activity through a predicted vitamin D response element, thereby promoting TG hydrolysis in cyp27b1[-/-] ovaries. In contrast, insufficient 1,25(OH)2D3 production increased Pparγ expression and promoted abnormal lipid deposition. Exogenous 1,25(OH)2D3 treatment reduced lipid deposition and restored Pparγ and Pnpla2 expression toward normal levels. In conclusion, our findings suggested that Cyp27b1 deficiency impaired ovarian development through lipid overload associated oxidative damage. These results linked impaired Cyp27b1 mediated 1,25(OH)2D3 production to ovarian lipid metabolic disturbance and redox imbalance. Lipid metabolic changes involving pnpla2 and pparγ may represent potential regulatory targets for 1,25(OH)2D3 deficiency associated ovarian pathologies.

Postmenopausal osteoporosis (PMOP) is frequently complicated by iron accumulation, which exacerbates bone loss. Baicalein, a natural flavonoid with iron-chelating and antioxidant properties, has shown bone-protective effects, but its role in iron-overloaded PMOP remains unclear. This study investigated whether baicalein ameliorates iron-accumulated PMOP by inhibiting osteoclastogenesis via Nrf2 activation. In vitro, bone marrow-derived macrophages (BMMs) were induced into osteoclasts with RANKL under iron overload. Baicalein (10 μM) suppressed osteoclast differentiation, F-actin ring formation, and bone resorption, reduced intracellular and mitochondrial ROS, lowered Fe[2+] and total iron content, and upregulated Nrf2 nuclear translocation and HO-1 expression. These effects were abrogated by the Nrf2 inhibitor ML385 or Nrf2 siRNA. In vivo, an ovariectomized (OVX) mouse model with iron dextran injection (100 mg/kg/week) was used. Oral baicalein treatment (100 mg/kg/day) significantly prevented bone loss, improved bone microarchitecture and biomechanical properties, reduced iron deposition in bone, liver and spleen, lowered serum ferritin and bone ROS, and activated Nrf2 signaling in bone tissue. Co-administration of ML385 reversed these protective effects. Collectively, baicalein ameliorates iron-accumulated postmenopausal osteoporosis by activating the Nrf2 pathway, thereby inhibiting osteoclast differentiation and bone resorption. Baicalein represents a promising multi-target candidate for treating PMOP, especially in patients with concomitant iron accumulation.

Obesity triggers ovarian dysfunction and female infertility accompanied by granulosa cell damage. Ferroptosis, iron-dependent lipid peroxidation-mediated cell death, participates in multiple reproductive diseases, yet the molecular cascade linking metabolic obesity to granulosa ferroptosis remains undefined. Using high-fat diet obese mice, primary human granulosa cells from obese infertile patients, and KGN cell lines, we identified the nuclear receptor NR4A1 as a key mediator of obesity-induced ferroptosis downstream of PI3K/AKT signaling. Elevated NR4A1 was associated with upregulated ACSL4 and suppressed GPX4, accompanied by iron overload, lipid peroxidation, and mitochondrial collapse, impairing folliculogenesis and oocyte quality. Metformin reduced NR4A1 activation in association with PI3K/AKT signaling modulation, thereby attenuating ferroptotic injury and contributing to the restoration of ovarian function. In vivo NR4A1 agonist administration attenuated the protective effects of metformin, while ovarian-specific NR4A1 knockdown alleviated HFD-triggered ovarian ferroptosis. Single-cell RNA-seq further suggested that metformin remodels oocyte redox and folliculogenic transcriptional networks. These findings support a role for the PI3K/AKT-NR4A1 signaling pathway in driving ferroptosis during obesity-related ovarian dysfunction and suggest a potential anti-ferroptotic mechanism of metformin in this context.

Growing evidence highlights the central role of inflammation in the onset and progression of DKD. In this study, we investigate the association between the neutrophil percentage-to-albumin ratio (NPAR) and clinicopathological features, as well as long-term renal outcomes, in patients with diabetic kidney disease (DKD). Three hundred five biopsy-confirmed DKD patients with baseline eGFR ≥ 15 mL/min/1.73 m[2] were retrospectively enrolled in this study. Baseline clinical and pathological characteristics were compared across NPAR quartiles. The primary outcome was 2-year DKD progression, defined as a > 40% increase in serum creatinine, end-stage renal disease, or all-cause mortality. Multivariable logistic regression, restricted cubic spline (RCS) analysis, and subgroup analyses were conducted to examine the association between the NPAR and kidney outcomes. Subsequently, a prognostic nomogram was constructed for DKD patients incorporating NPAR to stratify individual risk. Higher baseline NPAR quartiles were significantly associated with renal dysfunction and advanced renal pathological lesions. In a subset of renal biopsy specimens, MPO immunostaining demonstrated increased renal neutrophil infiltration in patients with higher NPAR quartiles. Renal MPO expression was positively correlated with mesangial expansion, interstitial fibrosis, and tubular atrophy. Among patients with complete follow-up, 65.3% developed the composite kidney outcome. Compared with the lowest NPAR quartile, multivariable-adjusted odds ratios (95% CIs) for kidney outcomes were 1.68 (0.36-8.15), 6.21 (1.35-33.55), and 6.88 (1.27-46.86) for Q2, Q3, and Q4, respectively. RCS analysis indicated a nonlinear association between NPAR and kidney outcomes, with risk increasing sharply above an NPAR value of 1.38. Subgroup analyses identified significant interactions between NPAR and Kimmelstiel-Wilson nodules as well as SGLT2 inhibitor use on kidney outcomes. A prognostic model incorporating NPAR quartiles, age, total cholesterol, eGFR, and interstitial fibrosis demonstrated good discrimination (AUROC 0.863). In conclusion, elevated NPAR is associated with increased renal neutrophil infiltration, more severe histopathological injury, and adverse renal outcomes in patients with DKD, and independently predicts long-term renal progression.