Adipose-derived mesenchymal stem cells (ADSCs) have demonstrated significant therapeutic effects on acute lung injury. Numerous studies have reported that preconditioning ADSCs can enhance their therapeutic efficacy. Currently, there is a lack of research on intermittent hypoxia preconditioning of ADSCs. In this study, we subjected ADSCs to intermittent hypoxic preconditioning, followed by the detection of their expressions of HIF-1α, HGF, VEGF, and IL-10. We also evaluated the therapeutic effects of ADSCs on oxidative stress and cell apoptosis in LPS-induced MLE-12 cells, and compared the outcomes resulting from different types of hypoxic preconditioning. Finally, we concluded that Intermittent hypoxia preconditioning leads to decrease in the secretion of VEGF by ADSCs compared to sustained hypoxia preconditioning; but their therapeutic effects in terms of anti-apoptosis and anti-oxidation are comparable.

Background: Transcranial alternating current stimulation (tACS) enhances motor performance, but its subcortical mechanisms in aged populations remain unclear. We investigated whether 20 Hz tACS over M1 engages striatal circuitry in aged mice under sevoflurane anesthesia via two parallel mechanisms: nigrostriatal dopamine release and THIN activation. Methods: Aged mice received 20 Hz tACS (0.15 mA, 20 min/day) or sham over M1 for seven days under sevoflurane. Motor performance was assessed by single‑pellet reaching task. Analyses included: FSCV for dopamine release; Golgi staining for spine density; c‑Fos/TH for THIN activation; EEG and cortical blood flow (CBF). Optogenetic gain‑of‑function with selective 20 Hz THIN activation was performed in a separate cohort. Results:tACS improved post‑anesthesia reaching success (p < 0.05), enhanced dopamine release (p < 0.01), THIN activation (p < 0.01), spine density (p < 0.01), and β‑band oscillations (p < 0.01). Optogenetic THIN activation recapitulated motor and β‑band enhancement (both p < 0.01), but not CBF increase. Conclusion: 20 Hz tACS enhances motor function in aged mice under sevoflurane via dopamine‑dependent plasticity and THIN‑mediated oscillatory modulation, identifying THINs as a key cellular substrate for tACS‑induced motor

Acute Lung Injury (ALI) and its severe manifestation, Acute Respiratory Distress Syndrome (ARDS), represent critical clinical challenges characterized by diffuse alveolar damage, uncontrolled inflammatory storms, and oxidative stress. Despite supportive therapies such as mechanical ventilation have advanced considerably, mortality rates remain persistently high. Over the past five years, Carbon Dots (CDs)-a novel class of zero-dimensional carbon nanomaterials-have demonstrated significant potential for the precision theranostics of ALI/ARDS due to their ultra-small size (< 10 nm), tunable photoluminescence, superior biocompatibility, and intrinsic enzyme-mimicking activities. This review comprehensively synthesizes frontier advancements in CDs applications for pulmonary diseases over the past five years. We systematically elucidate eco-friendly synthesis strategies, surface functionalization (eg, mannose and RGD peptide targeting), and the mechanisms by which CDs function as nanozymes (mimicking SOD, CAT, and POD) to scavenge reactive oxygen species (ROS). Particular emphasis is placed on novel therapeutic strategies, including the modulation of the gut-lung axis to remodel intestinal flora and the construction of ROS-responsive smart drug delivery systems (eg, for siRNA and glucocorticoids). Furthermore, we compare the inhalation toxicology of CDs against traditional carbon materials like carbon nanotubes and evaluate their utility in in vivo lung inflammation imaging and microenvironmental sensing (NO, pH). This review aims to provide a theoretical foundation and strategic direction for the clinical translation of CD-based nanomedicine.

Background Perioperative neurocognitive disorder (PND) is a frequent postoperative complication in older individuals and is commonly linked to microglial activation. Endoplasmic reticulum (ER) stress has been implicated in neuroinflammation and postoperative cognitive decline; however, whether hippocampal ER stress acts upstream to bias microglia toward a pro-inflammatory M1 phenotype, rather than serving only as a trigger of cytokine cascades, remains insufficiently defined in PND. To determine whether hippocampal ER stress promotes an M1-like microglial shift and amplifies neuroinflammation after anesthesia and surgery. Methods Eighteen-month-old male C57BL/6 mice underwent aseptic tibial fracture surgery under isoflurane anesthesia. Tauroursodeoxycholic acid（TUDCA）, a pharmacological ER stress suppressor, was delivered bilaterally into dorsal hippocampal CA1 as a perioperative mechanistic probe. ER ultrastructure in hippocampal cells was examined using transmission electron microscopy (TEM). ER stress related proteins (GRP78, p-PERK, p-eIF2ɑ, p-IRE1ɑ, ATF4, and ATF6) in hippocampus were quantified by Western blotting. Microglial phenotypic signatures were assessed by Western blotting (CD86, iNOS, CD206, ARG-1) and double immunofluorescence (CD86/Iba-1 and CD206/Iba-1). Open-field testing was performed to control for locomotion/anxiety, followed by memory assessments using novel object recognition and Morris water maze starting on postoperative day 3. Results Anesthesia and surgery induced postoperative cognitive deficits, accompanied by heightened hippocampal inflammatory signaling and a shift toward a pro-inflammatory microglial signature characterized by increased CD86 and iNOS with concomitant reductions in CD206 and ARG-1. In parallel, hippocampal ER stress activation was evident, including ER ultrastructural disruption and increased GRP78 with engagement of PERK/eIF2ɑ/ATF4-, IRE1ɑ-, and ATF6-related pathways. hippocampus-targeted delivery of TUDCA attenuated ER stress activation, mitigated the pro-inflammatory microglial shift (decreasing CD86/iNOS and restoring CD206/ARG-1), reduced inflammatory readouts, and improved postoperative memory performance. Conclusions These findings support a mechanistic link between hippocampal ER stress and the pro-inflammatory microglia bias linked to postoperative neuroinflammation and cognitive impairment in aged mice.

Hyperoxia-induced lung injury is a prominent inflammatory complication encountered in neonatal and adult critical care, contributing to acute lung injury and bronchopulmonary dysplasia. Although oxidative stress is a primary initiating factor, accumulating evidence suggests that dysregulated immune responses, particularly those mediated by macrophages, critically influence disease progression and resolution. Macrophages exhibit remarkable phenotypic plasticity in response to hyperoxic stress, extending beyond the conventional pro-inflammatory and anti-inflammatory polarization framework. Recent advances, including single-cell transcriptomic analyses, have revealed substantial heterogeneity among macrophage subsets, highlighting inflammatory, metabolically reprogrammed, senescent, and pyroptotic phenotypes in hyperoxic lung injury. These phenotypic shifts are tightly regulated by inflammatory signaling pathways, immunometabolic alterations, and cellular stress responses. In this review, we summarize current evidence regarding macrophage phenotypic plasticity in hyperoxia-induced lung injury, with a focus on key inflammatory pathways, metabolic reprogramming, inflammasome activation, and emerging cell fate programs such as pyroptosis and cellular senescence. We further discuss macrophage-mediated intercellular communication with epithelial and endothelial cells and its contribution to persistent inflammation and impaired lung repair. By integrating these findings, this review aims to provide updated insights into macrophage-driven inflammatory mechanisms and to highlight potential avenues for therapeutic modulation in hyperoxic lung injury.

Neuropathic pain (NP) is tightly linked to neuroinflammation driven by aberrant activation of spinal microglia; however, the endogenous regulatory mechanisms that limit excessive microglial responses following nerve injury remain incompletely understood. Adenosine (ADO) is widely recognized as a homeostatic mediator released under conditions of tissue damage and stress, and activation of the adenosine A1 receptor (A1R) has been reported to exert antinociceptive effects at both spinal and peripheral levels. Nevertheless, the potential role of A1R as an endogenous modulator of microglial activation in NP, as well as the underlying signaling mechanisms, remains poorly defined. In this study, we found that A1R expression was markedly increased in spinal microglia after nerve injury. Notably, ATPγS-mediated damage-associated molecular pattern (DAMP) stimulation, but not lipopolysaccharide (LPS)-mediated inflammatory stimulation, increased microglial A1R expression. Pharmacological activation of A1R reversed the pro-inflammatory phenotypic shift of microglia and suppressed their excessive proliferation, resulting in significant attenuation of NP behaviors across different injury models. Mechanistically, A1R activation inhibited extracellular signal-regulated kinase 1/2 (ERK1/2) phosphorylation in microglia, thereby preventing activation and nuclear translocation of the nuclear factor-κB (NF-κB) p65 subunit, reducing the release of pro-inflammatory cytokines (PICs) and alleviating the neuroinflammatory microenvironment in the spinal dorsal horn. Taken together, our findings suggest that injury-associated upregulation of microglial A1R may act as a negative feedback mechanism that restrains microglial activation and neuroinflammation via the ERK/NF-κB signaling axis, providing mechanistic insight into endogenous neuroimmune regulation following nerve injury.

Ovarian cancer (OC) is a prevalent malignancy; however, the role of kinesin light chain 3 (KLC3) in OC remains unclear. The present study conducted a comprehensive investigation of KLC3 using bioinformatics analysis, as well as in vitro and in vivo experiments. The findings revealed that KLC3 expression was significantly upregulated in the OC group compared with that in the normal group, and high KLC3 expression in patients with OC was associated with a poorer overall survival. Functional studies demonstrated that targeting KLC3 effectively suppressed the proliferation, migration, epithelial‑mesenchymal transition and DNA damage resistance of OC cells in vitro, while also inhibiting tumor growth in vivo, underscoring the pivotal role of KLC3 in tumor progression and metastasis. Additionally, RNA‑sequencing analysis identified collagen type III α1 (COL3A1) as a potential downstream gene cooperating with KLC3 to promote the occurrence and development of OC through the PI3K/AKT signaling pathway. Rescue experiments revealed that the KLC3 knockdown‑induced suppression of the malignant phenotype could be partially reversed by overexpression of COL3A1. In summary, the present findings demonstrated that KLC3 acts as an oncogene by influencing COL3A1 expression to promote the proliferation and migration of OC cells in vivo and in vitro.

Sepsis remains a major challenge in critical care, with high mortality despite ongoing improvements in treatment. The early uncontrolled burst of reactive oxygen and nitrogen species (RONS) and cytokine storms form a vicious cycle, ultimately leading to multiple organ dysfunction syndrome (MODS). The absence of effective therapies to interrupt this process is likely a key reason for poor outcomes. In recent years, the emergence of nanozymes has represented a transformative breakthrough in addressing this challenge. With strong antioxidant capacity, high stability, and low cost, nanozymes surpass conventional antioxidants and offer a promising therapeutic strategy for sepsis, especially through effective redox regulation. Nanozymes not only efficiently scavenge diverse RONS but also inhibit hyperactivated inflammatory pathways, thereby breaking the fatal vicious cycle between oxidative stress and cytokine storms. This provides a novel approach for immunomodulation and organ protection in sepsis. This review summarizes the key role of redox imbalance in sepsis progression and the therapeutic potential of nanozymes targeting redox imbalance, discusses their in vivo metabolic distribution and biosafety, and outlines prospects for future clinical translation and development. The objective is to provide insights that facilitate the development of innovative therapies targeting the RONS-inflammation axis in sepsis.

BackgroundTargeted therapy is a potent strategy for the treatment of advanced and metastatic cancers, with pancreatic cancer (PC) being one of the leading causes of cancer-related deaths worldwide. In order to resolve the limitations of existing targeted agents, there is an urgent need to find new targets and therapeutic strategies. Poly (ADP-ribose) polymerase 9 (PARP9) is aberrantly expressed in a variety of tumors. However, its relationship with PC has not been fully investigated. Lysyl oxidase like 2 (LOXL2) is potential therapeutic targets in challenging PC, which contributes to the malignant progression of PC and poor prognosis.MethodsCell lines with PARP9 knockdown or overexpression were established by lentiviral transfection, while LOXL2 was overexpressed by plasmid, and we validated the effect of PARP9 on apoptosis and DNA damage in PC cells using flow cytometry, comet assay, and western blots. The changes in downstream targets and signaling pathways after PARP9 expression changes were analyzed by RNA sequencing and protein blotting analysis. Finally, the relationship between PARP9 and LOXL2 was analyzed by immunoprecipitation, and the multidrug efflux system was determined by boinformatics analysis and western blots.ResultsPARP9 and LOXL2 were highly expressed in PC tissues and were associated with poor prognosis. PARP9 knockdown significantly inhibited the proliferation, invasion, and migration of PC cells, while also promoting apoptosis, increasing DNA damage, and inhibiting multidrug efflux systems. Meanwhile overexpression of LOXL2 reduced apoptosis and DNA damage, and drug sensitivity in PC cells caused by PARP9 knockdown. The therapeutic process of PARP9 in PC may be achieved through the synergistic action of LOXL2 and PI3K/AKT signaling pathway.ConclusionOur study reveals a potential link between PARP9 and PC, and targeting PARP9 and LOXL2 in monotherapy or combination therapy may provide novel strategies to advanced PC.

The serotonin (5-hydroxytryptamine) system represents a crucial neurotransmitter network that regulates mood, behavior, and cognitive functions, playing a significant role in the pathogenesis and progression of depression. Although this perspective faces significant challenges, the serotonin system continues to exert substantial modulatory effects on specific aspects of psychological functioning and actively contributes to multiple pathological processes in depression development. Therefore, this review systematically integrates interdisciplinary research advances regarding the relationship between the 5-hydroxytryptamine (5-HT) system and depression. By focusing on core biological processes including serotonin biosynthesis and metabolism, SERT gene regulatory networks, and protein molecular modifications, it aims to elucidate how 5-HT system dysregulation contributes to the development of depression, while providing novel research perspectives and therapeutic targets for innovative antidepressant drug development.

Targeted therapy is an effective strategy for the treatment of advanced and metastatic pancreatic cancer, one of the leading causes for cancer-related death worldwide. To address the limitations of existing targeted drugs, there is an urgently need to find novel targets and therapeutic strategies. Transcription factor FOS like 1 (FOSL1) is a potential therapeutic target for challenging pancreatic cancer, which contributes to the malignant progression and poor gnosis of pancreatic cancer. High mobility group A1 (HMGA1) is a nonhistone chromatin structural protein that contributes to malignant progression and poor prognosis of cancer. Human FOSL1 complete RNA, shRNA against FOSL1 and shRNA against HMGA1 lentiviral recombination vectors were used to overexpress FOSL1 and knock down FOSL1 and HMGA1. RNA sequencing, Q-PCR and Western blots were used to investigate the mechanism of FOSL1 in regulating the proliferation of pancreatic cancer cells. The relationship between FOSL1 and HMGA1 were analyzed by co-immunoprecipitation Mass spectrometry, Q-PCR of chromatin immunoprecipitation and Western blots. The regulation of FOSL1 and HMGA1 in the invasion and migration, stemness, and multidrug efflux system were determined by transwell assay, sphere formation assay, immunofluorescence, Q-PCR and Western blots. We found that FOSL1 promoted the proliferation and progression of pancreatic cancer by trigging stemness, invasion and metastasis, and drug resistance. HMGA1 was a key downstream target regulated by FOSL1 at the transcriptional level and directly interacted with FOSL1. Knockdown of HMGA1 inhibited the proliferation of pancreatic cancer cells by regulating the expression of genes related to stemness, epithelial-mesenchymal transition and multidrug efflux system. Targeted inhibition of FOSL1 and HMGA1 expression significantly inhibited the proliferation of pancreatic cancer cells. FOSL1 promote the malignant progression of pancreatic cancer by promoting HMGA1 expression. Targeting FOSL1 and HMGA1 in monotherapy or combination therapy is a promising strategy for the treatment of advanced and metastasis pancreatic cancer.

Gastric cancer (GC) is one of the most common tumors; one of the reasons for its poor prognosis is that GC cells can resist normal cell death process and therefore develop distant metastasis. Cuproptosis is a novel type of cell death and a limited number of studies have been conducted on the relationship between cuproptosis-related genes (CRGs) in GC. The purpose of the present study was to establish a prognostic model of CRGs and provide directions for the diagnosis and treatment of GC. Transcriptome and clinical data of patients with GC were collected from The Cancer Genome Atlas and Gene Expression Omnibus datasets. Single sample gene set enrichment analysis (GSEA) and the randomized forest method were used to establish the prognostic model. Kaplan-Meier survival curve, receiver operating characteristics diagram and a nomogram were used to evaluate the reliability of the model. GSEA and gene set variation analysis (GSVA) were used to examine enrichment pathways between high and low risk groups. Finally, immunohistochemical analysis was used to examine ephrin 4 (EFNA4) expression in GC samples and determine the prognosis of patients with GC based on the expression pattern of EFNA4. A group of 7 predictive models (RTKN2, INO80B, EFNA4, ELF2, MUSTN, KRTAP4, and ARHGEF40) was established which were correlated with CRGs. This model can be used as an independent prognostic factor to predict the prognosis of patients with GC. GSEA and GSVA results indicated that high risk patients with GC were mainly associated with the enrichment of ANGIOGENESIS and TGF_BETA_SIGNALING pathways. Finally, EFNA4 expression in GC was significantly higher than that in normal tissues, and patients with GC and high EFNA4 expression exhibited improved prognosis. In conclusion, the prognosis model based on CRGs could be used as the basis for predicting the potential prognosis of patients with GC and provide new insights for the treatment of GC.

Despite being a lifesaving intervention for many conditions, supplemental oxygen therapy carries the risk of severe hyperoxia-induced lung injury (HLI). The lack of effective countermeasures drives the urgent need for novel therapeutic strategies and mechanistic research to improve clinical outcomes. Eicosapentaenoic acid (EPA), a marine-derived polyunsaturated fatty acid with anti-inflammatory, antioxidant, and immunomodulatory properties, has exhibited lung-protective effects in experimental models of chronic obstructive pulmonary disease and bronchial asthma, suggesting its potential clinical value. However, its effects on HLI remain unexplored. This study investigated the therapeutic potential of EPA in alleviating HLI and elucidated its underlying mechanisms. We established both cellular and animal models of HLI; using these models, we integrated network pharmacology analysis to evaluate the efficacy of EPA and explore its potential mechanisms of action. Results demonstrated that EPA significantly ameliorated hyperoxia-induced oxidative stress, inflammatory responses, glycolytic reprogramming, and apoptosis in BEAS-2B and MLE-12 cells in vitro. Network pharmacology analysis revealed that EPA likely targets the MAPK/NF-κB signaling pathway. Western blotting and immunofluorescence confirmed that EPA attenuated hyperoxia-induced cellular damage by inhibiting p38 MAPK phosphorylation and reducing NF-κB nuclear translocation. In vivo, EPA suppressed the p38 MAPK/NF-κB signaling pathway, reduced inflammation and metabolic reprogramming in lung tissue, and thereby alleviated hyperoxia-induced pathological damage. This study substantiates that EPA ameliorates HLI by modulating glycolytic reprogramming via the p38 MAPK/NF-κB signaling pathway in vitro and in vivo, thereby providing a potential therapeutic strategy and laying the groundwork for its clinical application in HLI management.

BACKGROUND:Immunocompromise is common in the intensive care unit (ICU) and is strongly associated with adverse outcomes. However, robust quantitative tools for assessing the severity of immunocompromise are lacking. We aimed to develop and validate a machine learning-powered immunocompromise score and its risk stratification based on common immunocompromise conditions and biomarkers in the ICU. METHODS:A single-centre retrospective study was carried out in two ICUs of an academic tertiary care center in China. Adult patients who were admitted to the ICU for at least three days were enrolled. Feature selection was performed via the Boruta algorithm. The primary endpoint was 28-day all-cause mortality, whereas secondary endpoints included septic shock, the use of special antimicrobial agents, the peak levels of interleukin-6 (IL-6), etc. Seven machine learning models were developed via 10-fold cross-validation. The predicted probabilities from the optimal model were used to define the Immunocompromise and Severity (ICS) Score. To rapidly obtain patient immunocompromise information, a simplified ICS score (SICS score) was developed based on LASSO-selected features from day 1. Additionally, secondary endpoints were used to validate the rationale of ICS and SICS scores, and 216 patients were included for temporal validation. RESULTS:A total of 1863 patients were included for Algorithm derivation, with 679 deaths (36.9%). Among the seven machine learning models, the XGBoost model using data from the first 3 ICU days showed the highest performance, defined as ICS score (AUC 0.887; sensitivity 0.896; specificity 0.723; accuracy 0.787), and its performance was superior to that of the APACHE II and SOFA (P < 0.001). The LASSO algorithm selected 5 key variables (age, organ failure, immune impairing diseases and treatments, and IL-6 > 100 pg/mL with lymphopenia < 0.8 × 109/L) on ICU Day 1, for the SICS score (AUC 0.851; sensitivity 0.83; specificity 0.72; accuracy 0.765). The ICS and SICS scores effectively stratified patients into low, moderate, and high-risk groups with similar mortality rates (2.6% vs. 2.8%; 24.2% vs. 24.4%; 69.9% vs. 69.6%). Notably, the ICS score identified a greater proportion of patients in both the low (31.4% vs. 19.6%) and high risk (42.6% vs. 36.6%) groups. Secondary endpoints were significantly correlated with higher risk stratification, supporting the rationality of ICS and SICS scores. An additional 216 patients were used for temporal validation, yielding an AUC of 0.854 for the ICS score and 0.845 for the SICS score. CONCLUSIONS:Derived from common immunocompromising conditions and biomarkers, the ICS and SICS scores and their risk stratification system provide an accurate and timely solution for identifying and stratifying immunocompromise in critically ill patients. This framework may facilitate early clinical decision-making and resource allocation.

Polycystic ovary syndrome (PCOS), an endocrine disorder emerging in adolescence and reproductive years, has been linked to glycolysis in prior studies, though the precise mechanistic role of glycolysis in its pathogenesis remains unclear. Therefore, this study sought to identify glycolysis-related biomarkers in PCOS and elucidate their regulatory mechanisms to provide novel therapeutic strategies. Utilizing publicly available datasets, biomarkers were identified via differential analysis, various PPI algorithms, and validation of expression patterns. Subsequent analyses included functional enrichment, tissue and cell-specific expression profiling, m6A modification site prediction, compound screening, molecular network construction, and molecular docking. RT-qPCR was performed on clinical samples for experimental validation. Two biomarkers, TXNIP and TGFBI, were identified and jointly enriched in “complement and coagulation cascades”. TXNIP showed elevated expression in tongue and endocrine cells, whereas TGFBI was highly expressed in placental and adipocyte tissues. TGFBI had 14 high-confidence m6A modification sites and TXNIP had 1 high-confidence m6A modification site. The identified regulatory networks included hsa-miR-6761-5p-TXNIP-PPARG and hsa-miR-6761-5p-TGFBI-RB1. Four key compounds—acetaminophen, bisphenol A, tetrachlorodibenzodioxin, and valproic acid—were prioritized, with molecular docking revealing strongest binding affinities between bisphenol A and both biomarkers (TXNIP: -5.9 kcal/mol; TGFBI: -13.1 kcal/mol). RT-qPCR validation in granulosa cells from PCOS patients confirmed significant upregulation of TGFBI and TXNIP, aligning with bioinformatics predictions. These findings suggest that TXNIP and TGFBI may serve as potential biomarkers associated with glycolytic dysregulation in PCOS, offering insights into the interplay between metabolic dysfunction and disease mechanisms.

Background Phenotypic switching of vascular smooth muscle cells (VSMCs) serves as a critical pathological basis for various cardiovascular diseases. This phenotypic transformation enables cells to regain proliferative, secretory, and migratory capabilities, triggering pathological vascular remodeling and subsequent disease development, including pulmonary arterial hypertension (PAH). PAH represents a common complication of congenital heart disease (CHD). Beyond early surgical correction of anatomical defects, effective therapeutic options remain limited. Consequently, there is an urgent need to elucidate the mechanisms underlying PAH pathogenesis and identify novel therapeutic strategies. In CHD, pulmonary arteries are chronically exposed to a high-flow, hyperoxic environment. This study hypothesizes that chronic hyperoxia—a previously overlooked factor—may significantly contribute to phenotypic switching and functional alterations in pulmonary arterial smooth muscle cells (PASMCs). Methods PASMCs were isolated from Sprague-Dawley rats through enzymatic digestion and cultured under hyperoxic conditions to establish an in vitro hyperoxia model(95% O₂/5% CO₂). Cells were divided into two primary groups: Normoxia group (N-group) and Hyperoxia group (H-group). The effects of hyperoxia on phenotypic switching were assessed by examining the expression of contractile markers α-SMA and SM22α, and synthetic marker OPN using qRT-PCR and Western blot analysis. To evaluate changes in cellular secretory capacity, MMP-2 expression was analyzed at both mRNA and protein levels. Cell proliferation and migration capacities were evaluated using CCK-8 assays and scratch wound healing assays, respectively. To investigate the regulatory role of the PI3K/AKT pathway, cells were treated with the PI3K inhibitor LY294002 and divided into four experimental groups: Normoxia (N-group), Hyperoxia (H-group), Hyperoxia + LY294002 (HI-group), Normoxia + LY294002 (NI-group). The phenotypic and functional modifications of PASMCs were assessed using qRT-PCR, Western blot analysis, CCK-8 proliferation assays, and scratch wound healing migration assays to investigate the regulatory involvement of the PI3K/AKT signaling pathway. Results In PASMCs cultured under hyperoxic conditions, a phenotypic transition from contractile to synthetic state was observed. The expression of contractile proteins α-SMA and SM22α was downregulated, while the synthetic protein OPN was upregulated. Concurrently, hyperoxia induced elevated secretion of MMP-2, enhanced cellular viability as demonstrated by CCK-8 assay, and significantly augmented migratory capacity observed in wound healing assays. These alterations exhibited a time-dependent progression. Hyperoxia induced increased phosphorylation of PI3K and AKT, leading to activation of the PI3K/AKT signaling pathway. This activation was associated with enhanced cellular anti-apoptotic capacity and concomitant phenotypic and functional alterations Notably, treatment with the PI3K inhibitor LY294002 effectively abolished the hyperoxia-induced effects on both molecular profiles and cellular behavior. Conclusion The PI3K/AKT signaling pathway mediates hyperoxia-induced phenotypic transition in PASMCs, conferring enhanced proliferative, secretory, and migratory capacities. These findings establish a mechanistic link between chronic hyperoxia and pathological vascular remodeling, providing a novel pathological basis for CHD-associated PAH (CHD-PAH) development.

Background:Oxygen supplementation is essential for patients with a multitude of diseases but can cause severe hyperoxia-induced lung injury (HLI), necessitating the identification of therapeutic targets to improve clinical outcomes. Cuproptosis, a novel copper-dependent form of cell death characterized by proteotoxic stress resulting from lipoylated protein aggregation and loss of iron-sulfur cluster proteins, is distinct from other forms of cell death. However, the role of cuproptosis in HLI remains unclear. Methods:We established an HLI model in MLE-12 cells and C57BL/6 mice to investigate the involvement of cuproptosis in hyperoxia-induced toxicity. Results:We observed a time-dependent increase in the cuproptosis-related gene Fdx1 under hyperoxia. Moreover, hyperoxia activated the membrane-associated copper transporter SLC31A1 and significantly elevated copper levels in MLE-12 cells, as well as in the serum and lung tissue of C57BL/6 mice. Further analysis revealed that hyperoxia significantly altered the expression of cuproptosis-related genes without affecting DLAT levels, but significantly increased lipoylated-DLAT levels. ELISA, CCK-8 assays, HE staining, lung wet-to-dry weight ratio, and bronchoalveolar lavage fluid analysis demonstrated that treatment with the cuproptosis inhibitor TTM reduced pro-inflammatory cytokines (TNF-α and IL-1β) and alleviated hyperoxia-induced injury in both MLE-12 cells and C57BL/6 mice. Conclusion:Our study identifies the involvement of cuproptosis in HLI, providing new insights into the pathogenesis of hyperoxic lung injury and potential therapeutic strategies.

Hyperoxia therapy is a critical clinical intervention for both acute and chronic illnesses. However, prolonged exposure to high-concentration oxygen can cause lung injury. The mechanisms of hyperoxic lung injury (HLI) remain incompletely understood, and current treatment options are limited. Improving the safety of hyperoxia therapy has thus become an urgent priority. Ferroptosis, a novel form of regulated cell death characterized by iron accumulation and excessive lipid peroxidation, has been implicated in the pathogenesis of HLI, including diffuse alveolar damage, vascular endothelial injury, and bronchopulmonary dysplasia. In this review, we analyze the latest findings on ferroptosis and therapeutic strategies for HLI. Our aim is to provide new insights for the treatment of HLI and to facilitate the translation of these findings from bench to bedside.

Among the 321 million surgeries performed globally each year, sevoflurane dominates the inhaled anesthesia field due to its unique pharmacological properties. However, studies indicate that sevoflurane exerts multiple adverse effects on the nervous system, and its potential neurotoxic effects are increasingly drawing attention. This article integrates multi-level evidence from molecular mechanisms, cellular models, animal experiments, and clinical studies to comprehensively elucidate the key mechanisms underlying sevoflurane-induced neurotoxicity, including ferroptosis pathway activation, calcium homeostasis disruption, BDNF signaling abnormalities, neuroinflammatory responses, and endoplasmic reticulum stress. The findings aim to provide a theoretical foundation for developing precise neuroprotective strategies and optimizing clinical anesthesia protocols.