The rapid and accurate identification of nucleic acids (NAs) is essential to early diagnosis and treatment of disease in healthcare. Paper-based analytical devices (PADs) offer a preferred alternative to laborious, expensive, and time-consuming conventional nucleic acid testing (NAT) methods with considerable success. However, their development has been constrained by challenges in NA recognition and signal amplification within a restricted paper fiber network. Recently developed PADs, which establish a three-dimensional environment for NA reactions by the integration of porous fibers and modified substrate materials, are limited by the precision of base identification and user convenience. In this review, we focus on paper-based analytical devices with enhanced point-of-care applicability for NA detection and discuss the strategies for enhanced NA cascade signal amplification and background noise suppression. Emphasis is placed on the integrated detection of NA molecules from input to output, highlighting the effective signal amplification and rapid signal output of low-concentration NA molecules of the paper-based assay in complicated detection systems.

Distinguishing benign from malignant pulmonary nodules remains a major clinical challenge, where misdiagnosis may lead to either delayed cancer treatment or unnecessary invasive procedures. Here, we report apurinic/apyrimidinic endonuclease 1 (APE1)-triggered inhalable microsphere (ATIM) for non-small cell lung cancer (NSCLC) theranostics by leveraging inhalation delivery and homotypic targeting to accelerate the local enrichment of DNA tetrahedrons (TDNs) in pulmonary tumors, which enables in situ NSCLC theranostics. Upon intracellular recognition of the APE1, entropy-driven catalytic circuits are activated, triggering nanoparticle aggregation to amplify fluorescence for real-time tumor imaging and releasing miR-126-3p to induce tumor cell apoptosis by suppressing ADAM9. In a mouse orthotopic NSCLC model, the tumor-bearing group showed a fluorescent intensity 1.67-fold higher than the healthy group, and the pulmonary accumulation of the ATIM system via inhalation was 3.12-fold higher than that via intravenous injection, while ATIM therapy significantly reduced the tumor burden to a relative area of 45.3 ± 1.6%. Our results demonstrate that ATIM achieves accurate discrimination between benign and malignant pulmonary nodules while effectively inducing apoptosis in tumor cells. This theranostic system offers a promising dual-functional platform for precision diagnosis and targeted therapy in early-stage NSCLC.

The PD-1/PD-L1 blockade is a cornerstone of breast cancer immunotherapy, yet predictive biomarkers for immunotherapy outcomes remain scarce, restricting its clinical benefit. Glycosylated PD-L1 (gPD-L1) regulates tumor progression, immune suppression, and immune-checkpoint blockade (ICB) response, but it remains undetectable by conventional immunohistochemistry (IHC) due to glycan-mediated epitope shielding. This "detection blind spot" largely undermines current ICB biomarker prediction. To address this unmet clinical need, we developed an ultrasensitive CAP-g bioplatform with a novel CF/DPEDOT:PSS/Au nanoflexible scaffold. It integrates a dual-recognition-driven proximity cascade amplification for accurate gPD-L1 on small extracellular vesicle (gPD-L1-sEV) detection. We constructed this three-dimensional (3D) CF/DPEDOT:PSS/Au nanonetwork scaffold electrode, where π-π stacking synergistically minimizes charge-transfer resistance to improve electron transfer efficiency and maximizes the effective reaction area. Co-anchoring of a PD-L1 aptamer and a glycan probe further triggers hybridization chain reaction (HCR) for signal amplification, and this integrated design outperforms conventional electrochemical or immunological detection strategies. The CAP-g platform achieves an ultralow limit of detection (LOD) of 52 particles/mL, which is 1-2 orders of magnitude lower than existing gPD-L1-sEV or PD-L1-sEV detection methods, and a broad linear dynamic range of 103-108 particles/mL. Clinical validation in breast cancer patients and healthy controls showed robust discrimination between the two groups (P < 0.0001), with high specificity and reproducibility. This work overcomes the technical bottleneck of glycosylation interference in gPD-L1 detection, establishes a new liquid-biopsy paradigm for gPD-L1-sEV detection, and provides a reliable tool for predicting ICB efficacy in breast cancer, thus laying a critical foundation for personalized immunotherapy.

Acute kidney injury-induced acute lung injury (AKI-ALI) is a severe clinical syndrome characterized by systemic inflammation, oxidative stress, and immune cell activation. Vascular non-inflammatory molecule-1 (Vanin-1, VNN1), a pantetheinase enzyme involved in oxidative stress and inflammation, has been implicated in various inflammatory diseases. However, its role in AKI-ALI and its therapeutic potential remain unclear. An AKI-ALI model was established via bilateral kidney ischemia-reperfusion (KIR) in mice. VNN1-/- mice and pharmacological inhibition of Vanin-1 with RR6 were used to evaluate its role in AKI-ALI. Lung injury, oxidative stress, and inflammation were assessed using histological analysis, biochemical assays, and proteomic profiling. Neutrophil extracellular traps (NETs) formation was evaluated in vitro using immunofluorescence and ELISA. KIR-induced AKI resulted in severe lung injury, characterized by impaired oxygenation, increased broncho-alveolar lavage fluid protein leakage, and elevated inflammatory cytokines. Vanin-1 knockout significantly alleviated lung injury, reduced oxidative stress, and suppressed inflammation, without affecting renal injury. Proteomic and bioinformatics analyses revealed the pivotal role of neutrophils and their associated inflammatory responses during AKI-ALI. In vitro, Vanin-1 stimulation enhanced neutrophil activation and NETs formation. Pharmacological inhibition of Vanin-1 with RR6 significantly improved oxygenation, reduced lung injury, and attenuated oxidative stress and inflammation in AKI-ALI mice. Vanin-1 contributes to AKI-ALI progression by promoting NETs formation, oxidative stress, and inflammation. Both genetic deletion and pharmacological inhibition of Vanin-1 effectively alleviated lung injury, highlighting Vanin-1 as a promising therapeutic target for AKI-ALI.

Chronic diabetic wounds complicated by methicillin-resistant Staphylococcus aureus (S. aureus) (MRSA) remain refractory because the milieu evolves from a protease- and bacteria-dominated inflammatory phase to a reactive oxygen species (ROS)-rich, angiogenesis-impaired reparative phase. Here we report a stage-adaptive hydrogel dressing that integrates orthogonal, pathology-stage-driven activation with pathological-factor neutralization coupled to triggered delivery. Two bioresponsive microspheres are embedded in the hydrogel: MMP-9-responsive gelatin methacrylate (GelMA) microspheres loaded with a cationic antimicrobial peptide (ILRWPWWPWRRK-NH2), and ROS-responsive HA-PBA-PVA microspheres loaded with vascular endothelial growth factor (VEGF). In the early stage, elevated MMP-9 cleaves GelMA microspheres, consuming MMP-9 and releasing AMP to eradicate MRSA and mitigate infection-driven inflammation. In the later stage, ROS degrades HA-PBA-PVA microspheres, quenching ROS while releasing VEGF to restore endothelial function and neovascularization. In vitro, the system shows stimuli-responsive release, potent antibacterial activity, ROS scavenging, and improved endothelial migration and tube formation under oxidative stress. In a streptozotocin-induced diabetic rat model with MRSA-infected full-thickness wounds, the dual-microsphere hydrogel accelerates closure, reduces bacterial burden and inflammation, enhances collagen deposition and mature vessel formation, and shows no systemic toxicity. This work demonstrates a translational, stage-matched strategy for regulating infected diabetic wounds.

Precise imaging of cancer cells serves as the foundation for subtype analysis, significantly advancing the development of precision medicine. The unavoidable cellular internalization of fluorescent labels constrains the resolution and timeliness, presenting a significant obstacle. This study introduces a dynamic decorating strategy of DNA scaffolds that enables the execution of four distinct Boolean logic operations through self-assembly and self-disassembly. By integrating the in-built molecular circuit, the proposed assay achieved signal-amplified detection of low-abundance nucleic acid inputs. In the application of cell imaging, inputs were labeled with aptamers to operate membrane-confined self-assembly or self-disassembly of DNA scaffolds on the cell surface, enabling simultaneous identification of distinct subtypes of cancer cells with high fidelity. The intrinsic durability of DNA scaffolds successfully prevented cellular internalization for up to 300 min, boosting the long-duration imaging. Moreover, the assay was capable of profiling a broad spectrum of cancer cell abundances from as low as 0.1% to 10% in clinical blood samples, consistently achieving recognition efficiency exceeding 60%. These findings underscore the transformational potential of DNA scaffold-based imaging tools in biological research and precision medicine.

Background:Sepsis is a life-threatening condition caused by a dysregulated host response to infection, characterized by biphasic immune dysregulation and high mortality rates. Artesunate (AS), a semisynthetic artemisinin derivative, has demonstrated broad pharmacological properties, yet its overall efficacy and mechanisms in sepsis remain systematically unassessed at the preclinical level. Objectives:In this study, we aimed to conduct the first systematic review and meta-analysis to evaluate the therapeutic efficacy and underlying mechanisms of AS in animal models of sepsis. Methods:We systematically searched five electronic databases up to 3 September 2025, for controlled in vivo studies analyzing the effects of AS in septic animals. The study quality was assessed using the SYRCLE risk-of-bias tool, and evidence certainty was rated via the GRADE approach. Statistical analyses, including meta-analysis, publication bias, and sensitivity analyses, were performed using RevMan 5.4 and Stata 17.0. Results:Fifteen studies involving mice and rats were included. Meta-analysis indicated that AS was associated with improved survival (10 studies, OR: 6.87, 95% CI: 3.81-12.41, p < 0.00001), reduced bacterial load, and promotion of body weight recovery. Organ protection was evidenced by attenuated lung injury (reduced histological scores, MPO activity, and wet-to-dry ratio) and improved liver function (decreased AST and ALT levels). Analysis of cytokine data from different time-points suggested a potential phase-dependent immunomodulatory effect: AS suppressed pro-inflammatory cytokines (TNF-α and IL-6) during the hyperinflammatory phase while restoring immune competence in the immunosuppressive phase, accompanied by elevated IL-1β. Furthermore, AS reduced apoptosis (decreased TUNEL-positive cells) and enhanced pro-survival signaling (increased p-mTOR/mTOR ratio); however, its effect on caspase-3 was not significant. Sensitivity analyses supported the robustness of the primary findings, and no significant publication bias was detected within the limits of the available studies. Conclusion:AS is associated with survival benefits and multi-organ protection in septic animal models through multimodal mechanisms, potential phase-aware immunomodulation, antiapoptotic effects, and enhanced bacterial clearance. Despite methodological heterogeneity across studies, these preclinical findings support further investigation of AS as a potential therapeutic candidate for sepsis treatment. Systematic Review Registration:https://www.crd.york.ac.uk/PROSPERO/view/CRD420251146068.

Enterovirus A71 (EV-A71) is a major causative agent of hand, foot, and mouth disease, yet no specific antiviral therapy has been approved. This study systematically evaluated the efficacy and mechanisms of alkaloids against EV-A71 infection by integrating meta-analysis, network pharmacology, and molecular docking. Nine animal studies were included. Meta-analysis suggested that alkaloid intervention significantly improved survival (OR = 30.62, 95% CI: 10.44-89.82), reduced clinical severity, attenuated body weight loss, and decreased viral loads in infected tissues. Subgroup analyses preliminarily suggested that quinolizidine alkaloids and high-dose regimens (>5 mg/kg) may be associated with preclinical intervention effects. Network pharmacology predicted 155 shared targets between seven active alkaloids and EV-A71-related genes, with MAPK1, MAPK3, JUN, AURKB, and MAPK8 recognized as core targets through computational screening. Functional enrichment analysis suggested significant involvement of the MAPK, TNF, and IL-17 signaling pathways. Molecular docking provided computational support for stable binding affinities between active alkaloids and core targets (-6.7 to -9.3 kcal/mol). Collectively, these findings suggest that alkaloids exert anti-EV-A71 effects through both direct antiviral activity and host-directed regulatory mechanisms, supporting their potential as candidates for the development of novel anti-EV-A71 therapeutics.

High-fidelity profiling of intracellular biomarkers is crucial for deciphering tumorigenesis and progression, as well as classifying cell types. Conventional techniques suffer from high background interference and poor stability due to the scattered, point-like signals. Herein, we present a DNA tile-driven in vivo logical encoding self-assembly (TILES) strategy for robust and signal-amplified imaging of multiple biomarkers in cancer cells and tumor-bearing mice. Capitalizing on AND-gated logic operations, the TILES assay enables confined recognition of dual biomarkers to initiate the intracellular self-assembly of DNA scaffolds, yielding nanotube-like signals. Owing to the compact nucleic acid framework, DNA scaffolds exhibit an over sevenfold nuclease resistance enhancement than double-stranded DNA probes. Moreover, the structural output, characterized by high-density fluorophore architecture and defined morphology, substantially boosts the visualization and precise classification of diverse cancer types. The proposed strategy provides a robust platform to enable the high-fidelity, multiplexed profiling of intracellular biomarkers and dynamic gene regulatory networks analysis in living organisms.

Background Preeclampsia (PE) is a pregnancy-related hypertensive disorder and a leading cause of maternal and perinatal mortality. Current treatments focus primarily on symptom management, as delivery remains the only definitive cure. This underscores the urgent need for innovative therapeutic strategies. Cytokines released by placental immune cells may contribute to the progression of PE and represent promising therapeutic targets. Methods We conducted single-cell sequencing on placental tissues obtained via cesarean section from patients with severe PE and cases of non-infectious preterm birth. Machine learning was applied to identify critical immune cells. Immune Response Enrichment Analysis was performed on these key cells to evaluate their polarization states and cytokine responses associated with PE. Additionally, we investigated cell–cell communications, key genes, and their related functions Results We identified cell type-specific alterations in PE, including an increased proportion of CD4+ T cells polarized towards an IL-1α- and IL-1β enriched T4-c state. Altered SPP1 signaling between macrophages and CD4+ T cells was observed, indicating immune response dysregulation. Additionally, machine learning algorithms identified four hub genes, and six small-molecule drugs were predicted to hold therapeutic potential for PE. Conclusions This study emphasizes the importance of further understanding CD4+ T cell dynamics in preeclampsia, providing potential insights into the principles of placental immunity and contributing to the exploration of immunotherapeutic strategies ### Competing Interest Statement The authors have declared no competing interest.

Tumor‐associated macrophages (TAMs) are one of the most abundant immune cells in solid tumors and play a critical role in tumor progression. This study found that the expression of LIM domain‐only protein 7 (LMO7) in TAMs is associated with poor patient survival. LMO7 deficiency significantly inhibited tumor growth and increased the accumulation of antitumor TAMs and CD8 + T cells. Specifically, single‐cell RNA sequencing (scRNA‐seq) reveals that LMO7‐deficient TAMs undergo an antitumor reprogramming, characterized by upregulated expression of pro‐inflammatory and phagocytosis‐related genes. Notably, LMO7 deficiency enhances immune‐mediated tumor confinement by regulating the phagocytic activity of TAMs. Mechanistically, LMO7 inhibits TAM phagocytosis by promoting the lysine 48‐linked polyubiquitination at lysine 45 of the β chain of the phagocytic receptor low‐density lipoprotein receptor‐related protein 1 (LRP1), leading to degradation via the ubiquitin‐proteasome system. Furthermore, combined targeting of LMO7 deficiency and SIRPα blockade demonstrates synergistic antitumor efficacy. Collectively, these findings demonstrate the critical role of LMO7 in orchestrating TAM phagocytosis and suggest that LMO7 inhibition is a promising drug target to enhance cancer immunotherapy.

INTRODUCTION:The intricate balance between immunometabolic homeostasis and redox equilibrium is crucial for maintaining health, and its dysregulation is implicated in a wide spectrum of diseases. Vascular non-inflammatory molecule-1 (VNN1) is an emerging pantetheinase that sits at the crossroads of inflammation and metabolism, yet a comprehensive review that synthesizes its tissue- and disease-specific roles and systematically evaluates its potential as a therapeutic target remains lacking. METHODS:A systematic literature search was conducted to identify relevant domestic and international studies on VNN1. The search included databases such as PubMed using keywords related to VNN1's structure and function, the disease roles of its metabolites (pantothenic acid, cysteamine), or inhibitors efficacy. The selected studies were critically reviewed and summarized to extract key pathways, inhibitor profiles and molecular docking analyses synthesized. RESULTS:VNN1 hydrolyzes pantetheine to generate metabolites essential for CoA synthesis and glutathione redox balance. Its upregulation is closely associated with the pathogenesis of acute and chronic inflammatory diseases and certain cancers, often serving as a biomarker for disease severity. Inhibiting VNN1, either genetically or pharmacologically with compounds like RR6, OMP-7, or natural products such as oleuropein, demonstrates significant anti-inflammatory and antioxidant effects in preclinical models. CONCLUSIONS:VNN1 represents a promising therapeutic target for modulating oxidative stress and immunometabolism in various diseases. Future research should develop disease-specific inhibitors, clarify tissue-specific mechanisms, and conduct clinical trials for translation.

The absence of sensitive, multiplexed, and point-of-care assays poses a critical obstacle in promptly responding to emerging human respiratory virus (HRV) pandemics. Herein, RECOGNIZER (re-building commercial pregnancy strips via large-size nanoflowers), an innovative one-pot CRISPR assay, is presented that employs commercially available strips to identify several types of HRVs. The superiority of the RECOGNIZER assay mainly relies on two aspects: (i) DNA nanoflowers possessing a high surface-to-volume ratio and well-defined surface allow for a considerable probe loading density and minimized non-specific interaction, achieving an impressive signal-to-noise proportion exceeding tenfold at 1 nM target. (ii) The design of the one-pot reaction, multi-channel chip, and custom-made app enables the rapid, sample-to-answer, and multiplexed analysis of four HRVs in 25 min. This assay demonstrates a sensitivity of 5.42 pM for synthetic SARS-CoV-2 RNA and 10 copies mu L-1 for SARS-CoV-2 plasmids after pre-amplification. Finally, the proposed approach indicated 100% accuracy in 50 clinical swab samples, demonstrating the robust performance in distinguishing SARS-CoV-2 from other HRVs. The versatility and scalability of RECOGNIZER renders it a user-friendly platform for virus infection monitoring, offering significant potential for improving pandemic response efforts.

Dynamic tracing of intracellular telomerase activity plays a crucial role in cancer cell recognition and correspondingly in earlier cancer diagnosis and personalized precision therapy. However, due to the complexity of the required reaction system and insufficient loading of reaction components into cells, achieving a high-fidelity determination of telomerase activity is still a challenge. Herein, an Aptamer-Liposome mediated Telomerase activated poly-Molecular beacon Arborescent Nanoassembly(ALTMAN) approach was described for direct high-fidelity visualization of telomerase activity. Briefly, intracellular telomerase activates molecular beacons, causing their hairpin structures to unfold and produce fluorescent signals. Furthermore, multiple molecular beacons can self-assemble, forming arborescent nanostructures and leading to exponential amplification of fluorescent signals. Integrating the enzyme-free isothermal signal amplification successfully increased the sensitivity and reduced interference by leveraging the skillful design of the molecular beacon and the extension of the telomerase-activated TTAGGG repeat sequence. The proposed approach enabled ultrasensitive visualization of activated telomerase exclusively with a prominent detection limit of 2 cells center dot mu L-1 and realized real-time imaging of telomerase activity in living cancer cells including blood samples from breast cancer patients and urine samples from bladder cancer patients. This approach opens an avenue for establishing a telomerase activity determination and in situ monitoring technique that can facilitate both telomerase fundamental biological studies and cancer diagnostics.

Enterovirus A71 (EV-A71) is a major pathogen causing hand, foot, and mouth disease (HFMD) in children worldwide. It can lead to severe gastrointestinal, pulmonary, and neurological complications. The innate immune system, which rapidly detects pathogens via pathogen-associated molecular patterns or pathogen-encoded effectors, serves as the first defensive line against EV-A71 infection. Concurrently, the virus has developed various sophisticated strategies to evade host antiviral responses and establish productive infection. Thus, the virus-host interactions and conflicts, as well as the ability to govern biological events at this first line of defense, contribute significantly to the pathogenesis and outcomes of EV-A71 infection. In this review, we update recent progress on host innate immune responses to EV-A71 infection. In addition, we discuss the underlying strategies employed by EV-A71 to escape host innate immune responses. A better understanding of the interplay between EV-A71 and host innate immunity may unravel potential antiviral targets, as well as strategies that can improve patient outcomes.

Sepsis is a syndrome of organ dysfunction caused by an uncontrolled inflammatory response, which can seriously endanger life. Currently, there is still a shortage of specific therapeutic drugs. Quercetin and its natural derivatives have received a lot of attention recently for their potential in treating sepsis. Here, we provide a comprehensive summary of the recent research progress on quercetin and its derivatives, with a focus on their specific mechanisms of antioxidation and anti-inflammation. To obtain the necessary information, we conducted a search in the PubMed, Web of Science, EBSCO, and Cochrane library databases using the keywords sepsis, anti-inflammatory, antioxidant, anti-infection, quercetin, and its natural derivatives to identify relevant research from 6315 articles published in the last five years. At present, quercetin and its 11 derivatives have been intensively studied. They primarily exert their antioxidation and anti-inflammation effects through the PI3K/AKT/NF-κB, Nrf2/ARE, and MAPK pathways. The feasibility of these compounds in experimental models and clinical application were also discussed. In conclusion, quercetin and its natural derivatives have good application potential in the treatment of sepsis.

Background: To assess the relationship between soluble interleukin-17 receptor (sIL-7R) levels and CD3-positive t cells and lymphocytes in patients with sepsis and their predictive clinical significance. Methods: The study cohort comprised individuals diagnosed with sepsis based on the Third International Consensus Definitions for Sepsis and Septic Shock, treated in the emergency and critical care medicine departments at Beijing Chuiyangliu Hospital and Baoding No. 1 Central Hospital between December 2020 and June 2022. Patient outcomes were classified based on survival or mortality. Biomarkers, including sIL-7R levels and illness severity scores, were documented. All statistical analyses, including predictive modeling and comparisons were carried out using SPSS v.23.0 software and R software. Results: On the fifth day post-admission, sIL-7R levels significantly decreased in both the survival and death groups, compared with levels on day one (2.09 +/- 0.65 vs 1.07 +/- 0.53 ng/mL, P < 0.01). There was a significant correlation between the sIL-7R level and the CD3+ T-lymphocyte count (CD3+) (r = 0.44) and lymphocyte count (LYM) (r = 0.42). The combination of the sIL-7R level with the Sequential Organ Failure Assessment (SOFA) score demonstrated optimal predictive value for clinical outcomes in patients with sepsis, demonstrated by an area under the receiver operating characteristic curve of 0.998. Conclusion: sIL-7R levels are correlated with CD3+ and LYM counts. Additionally, the combination of serum sIL-7R level and SOFA score provides a robust method for predicting sepsis outcomes.

Metastasis suppressor 1 (MTSS1) has been reported to play important roles in suppressing cancer progression. In this study, we investigated the underlying mechanism that regulates MTSS1 expression. We showed that in breast cancer cells, lncRNA-SNHG15-induced cell invasion and proliferation was accompanied with the decreased expression of MTSS1 mRNA. Further study revealed that SNHG15 mediated MTSS1 repression through blocking its promoter activity. Mechanistically, SNHG15 complexes with DDX5 and RTF1 and interacts with the core promoter of the MTSS1 gene to interfere with RNA-Pol-II-directed transcriptional initiation. Association with DDX5 stabilizes SNHG15 while binding to RTF1 allows SNHG15 to carry RTF1 to the core promoter, where RTF1 forms a complex with PNA pl II to enhance transcriptional pausing. Our findings revealed a molecular mechanism by which SNHG15 serves as a regulator to suppresses MTSS1 transcription via interaction with the gene core promoter.

BACKGROUND Atherosclerosis is a chronic lipid-driven inflammatory disease, largely influenced by hemodynamics. Neutrophil extracellular traps (NETs)-mediated inflammation plays an important role in atherosclerosis. However, little is known about the mechanism of the generation of NETs under different shear stress and subsequent damage to endothelial cells. We sought to identify a novel mechanical signal provokes NETs generation and to investigate its potential role in atherosclerosis. METHODS ApoE −/− mice were fed with high-fat diet (HFD) to induce atherosclerosis. The model of lower shear stress (LSS) with a partial ligation of the left carotid artery was established to assess the role of LSS in NETs generation and atherosclerotic lesions development. Furthermore, the underlying mechanism of LSS promoting NETs generation and injuring endothelial cells was deciphered in neutrophil-like human promyelocytic leukemia (HL-60) cells in parallel-plate flow chamber. RESULTS We found that LSS correlated spatially with both NETs and atherosclerosis, while inhibition of NETosis could significantly reduce plaque formation in ApoE −/− mice. In vitro , LSS could promote NETs generation directly through down-regulation of Piezo1, a mechanosensitive ion channel. downexpression of Piezol could activate neutrophils and promote NETosis in static. Conversely, Yoda1-evoked activation of Piezo1 attenuated LSS-induced NETosis. Mechanistically, the downexpression of Piezo1 resulted in decreased Ca 2+ influx and increased histone deacetylase 2 (HDAC2), which increase reactive oxygen species levels, then led to NETosis. LSS-induced NETs generation promoted the apoptosis and adherence of endothelial cells. CONCLUSIONS LSS directly promotes NETosis through piezo1-HDAC2 axis in atherosclerosis progression. This study uncovers the essential role of Piezo1-mediated mechanical signaling in NETs generation and plaque formation, which provides a promising therapeutic strategy for atherosclerosis. Graphic Abstract Proposed mechanism for lower shear stress LSS exacerbating atherosclerosis. LSS stimuli decrease Piezo1 expression in the neutrophils, resulting in decreased intracellular Ca 2+ concentration, as well as the higher expression level of HDAC2, which could activate oxidative stress and promote intracellular reactive oxygen species formation, and ultimately lead to NETs generation. NETs could aggravate endothelial cells injury and exasperate atherosclerosis. Highlights ■ Lower shear stress (LSS) promotes Neutrophil extracellular traps (NETs) formation, which is critical for lipid deposits and plaque formation in Atherosclerosis. ■ Atherosclerotic plaque formation was significantly reduced in the aorta of high fat diet fed ApoE −/− mice intraperitoneal injected with NETs inhibitor, GSK484, especially in the lower shear stress regions. ■ Piezo1 is a key molecule in the process of neutrophils sense lower shear stress. ■ lower shear stress inhibits the activation of Piezo1 and promotes NETosis through piezo1-HDAC2 axis. ■ LSS-induced NETs promote the apoptosis and adhesion of endothelial cells.