Ferroptosis, driven by iron-dependent lipid peroxidation, represents a promising therapeutic strategy for hepatocellular carcinoma (HCC). However, reactive oxygen species (ROS) released from ferroptotic HCC cells may activate hepatic stellate cells (HSCs), potentially exacerbating liver fibrosis, an unexpected risk in HCC treatment. Resolving this rapid intercellular crosstalk requires methods that can track extracellular ROS release and key parameters of HSC activation (i.e., intracellular redox homeostasis and mitochondrial activity) with high spatiotemporal resolution. Here, we established an in vitro coculture model of HuH7 (HCC) and LX-2 (HSCs) cells and employed scanning electrochemical microscopy (SECM) to spatiotemporally resolve extracellular ROS fluxes from ferroptotic HuH7 cells, while subsequent intracellular ROS generation, mitochondrial respiratory activity, and intracellular temperature changes in LX-2 cells. We found that ROS released from ferroptotic HuH7 cells increased by ∼2.44 μM, and was temporally followed by NOX2-associated intracellular ROS bursts in LX-2 cells, with a maximum generation rate of approximately 5.44 × 10-18 mol min-1. This intracellular ROS burst was coupled to mitochondrial hypermetabolism in LX-2 cells, reflected by a ∼1.27-fold elevation in oxygen consumption and a ∼1.65 K rise in intracellular temperature. Finally, we also found that pirfenidone, an antifibrotic agent, could effectively suppress this ferroptosis-induced HSC activation. Our study introduces an SECM approach providing real-time single-cell evidence that ferroptotic HCC cells can trigger ROS-mediated activation of neighboring HSCs, and supports further evaluation of antifibrotic combination strategies for safer ferroptosis-based HCC therapy.

ABSTRACT Hydrogel microfluidic devices have garnered significant interest as cell culture platforms owing to the efficient diffusion of nutrients through the hydrogel. Stereolithography, a versatile fabrication technique, has been widely adopted for the rapid prototyping of various microfluidic devices. Although evaluating cellular activity within microchannels during in vitro assays is essential, integrating electrochemical sensors into hydrogel devices remains challenging as these materials often cannot withstand conventional electrode fabrication processes. To address this limitation, scanning electrochemical microscopy was employed to assess cellular activity within hydrogel microfluidic devices, thereby eliminating the requirement for internal electrode fabrication. Initially, a novel hydrogel microfluidic design was developed, featuring accessible regions and thin hydrogel films. As a proof of concept, the respiratory activity of MCF‐7 cells within a hydrogel microchannel was measured. This strategy is expected to facilitate future cell‐based drug‐screening applications.

Liver cancer (LC) remains one of the most prevalent and lethal malignancies worldwide. Alpha-fetoprotein (AFP) and miRNA 106a are both robust biomarkers of LC, and the multiplexed detection of these two biomarkers can significantly improve the diagnostic accuracy over single-analyte strategies. Here, we present an extended gate indium-tin-zinc-oxide (ITZO) thin-film transistor (TFT) biosensor array that enables label-free, multiplexed detection of AFP and miRNA 106a, achieving a linear dynamic range spanning nine orders of magnitude and ultralow detection limits of 83.37 ag/mL and 35.48 aM, respectively. The extended gate physically decouples the biorecognition interface from the ITZO channel, preserving high transconductance while preventing ionic and biological degradation of the biosensor, greatly enhancing the biosensor's long-term stability with >98% of its initial sensitivity remained after one month of refrigerated storage. As a consequence, clinical detection results obtained from the biosensor array platform revealed that a dual-biomarker "OR" logic achieves 100% diagnostic accuracy (AUC = 1.00) when analyzing undiluted serum from LC patients and healthy donors. Therefore, the proposed platform offers a miniaturized, low-cost alternative for early LC screening and timely therapeutic intervention.

Human intestinal organoids serve as valuable models in regenerative medicine and drug screening. Nondestructive evaluation of both the hydrogel domes and the organoids cultured within them, including assessment of alkaline phosphatase (ALP), a key differentiation marker, remains a critical analytical challenge. To address this, we have developed a scanning electrochemical microscopy (SECM)-based approach that enables in-situ, real-time monitoring of ALP activity of organoids in hydrogel domes. Based on the recorded oxidation currents of enzymatic product of ALP, we have quantitatively determined ALP activity at the single-organoid level, reporting for the first time the enzyme activity in units per organoid (where 1 U corresponds to 1 μmol of product formed per minute). This innovative SECM strategy provides a noninvasive and spatially resolved analytical platform that is expected to advance organoid-based drug screening and transplantation studies.

Single-cell analysis is essential for probing cellular heterogeneity and understanding biological and disease mechanisms. Cellular imaging plays a key role in this field by providing critical spatiotemporal and functional information. Scanning electrochemical probe microscopy (SEPM), a kind of electrochemical imaging technique using micro/nanometer-sized probes to scan cells, can provide information on cellular topography and various interfacial processes in a nondestructive and real-time manner with high spatiotemporal resolution, therefore becoming a powerful tool for single-cell analysis. In this perspective, we systematically review the advances of three SEPM techniques (including scanning electrochemical microscopy (SECM), scanning ion conductance microscopy (SICM), and scanning electrochemical cell microscopy (SECCM) for single-cell analysis, including imaging cellular topography, membrane protein distribution, metabolic activity, surface charge, and mechanical property. We also discuss the current challenges in enhancing spatiotemporal resolution, detection sensitivity, and imaging throughput and realizing the multiparameter monitoring capability of SEPM for single-cell studies. Last, we highlight their promising development directions, such as high-performance imaging, multimodal integration, functionalized probes, 4D monitoring, and intelligent data analysis, with which SEPM can support more in-depth investigation in cell function studies.

Background:Patients with psoriasis often experience emotional distress, which may be alleviated through psychological interventions. Objective:To evaluate online Mindfulness-Based Cognitive Therapy (MBCT) as an adjuvant treatment for psoriasis, focusing on severity of lesion, anxiety, depression, quality of life, and itching. Methods:This randomized trial enrolled 109 patients with psoriasis assigned to either treatment as usual (TAU) or TAU plus MBCT, which included 8 weekly online sessions. Primary outcomes included Psoriasis Area and Severity Index (PASI), Self-Rating Anxiety/Depression Scale (SAS/SDS), and Dermatology Life Quality Index (DLQI). Itching was a secondary outcome measured by Visual Analogue Scale (VAS). Assessments were conducted at baseline, 4, 8, and 12 weeks. Treatment effects were analyzed with mixed linear models. Results:A total of 109 patients were randomized 1:1 to MBCT + TAU (n = 53) or TAU (n = 56). Significant group × time interactions favoring MBCT + TAU were observed for PASI (F = 3.746, P = .013, d = 0.331), SDS (F = 3.205, P = .025, d = 0.269), DLQI (F = 3.130, P = .028, d = 0.309), and VAS (F = 3.126, P = .028, d = 0.295). No significant between-group difference was found for anxiety. Limitations:Single-center. Conclusions:Adjunctive online MBCT led to improvements in psoriasis severity, depression, DLQI and itching, suggesting its potential to enhance comprehensive psoriasis management.

This study establishes a stable, reproducible, and biologically relevant system for the isolation, identification, and cultivation of primary keratinocytes from mouse tails, aiming to provide reliable in vitro models for investigating psoriasis pathogenesis and drug screening. Primary cells were isolated using a sequential digestion protocol involving Dispase II and trypsin, followed by purity validation via Krt14 immunofluorescence and flow cytometry. Using primary cells and human keratinocyte cell line (HaCaT), two-dimensional (2D) and three-dimensional (3D) psoriasis-like cell models were successfully established. The 2D model was induced with the M5 cocktail, a cytokine mixture consisting of TNF-α, IL-1α, IL-6, IL-17A and IL-22 to mimic the psoriatic inflammatory microenvironment, and utilized for assessing cell proliferation and inflammatory factor expression. The 3D model was generated using the air-liquid interface (ALI) culture technique, reconstructing a physiologically stratified epidermal structure, and its features were characterized by hematoxylin-eosin (H&E) staining after M5 stimulation to evaluate epidermal hyperplasia and differentiation. This dual-model system integrates the high-throughput capacity of the 2D platform with the physiological relevance of the 3D platform, offering an important experimental tool for deciphering the pathological mechanisms of psoriasis and screening candidate therapeutic agents.

Resveratrol (RES), a naturally occurring polyphenolic compound found in grapes, berries, and peanuts, has attracted considerable interest because of its antioxidant, anti−inflammatory, and neuroprotective properties. This narrative review examines the current evidence regarding the potential effects of RES on memory−related processes and neuroinflammatory biomarkers in major neurological disorders, including Alzheimer’s disease (AD), Parkinson’s disease (PD), multiple sclerosis (MS), and cerebral ischemia. Relevant literature was identified through searches of major scientific databases, and studies addressing the molecular mechanisms, experimental outcomes, and therapeutic implications of RES in these conditions were evaluated. The available evidence indicates that RES can modulate several biological pathways associated with neurodegeneration, including oxidative stress, inflammatory signaling, mitochondrial dysfunction, and neuronal survival. Experimental studies suggest that RES may influence key molecular mediators such as pro−inflammatory cytokines, nitric oxide (NO) signaling, and matrix metalloproteinases, which are implicated in neuronal damage and blood–brain barrier disruption. In preclinical models of AD and PD, RES has been associated with improvements in cognitive performance, reduction of neuroinflammatory markers, and attenuation of neuronal loss. Similarly, studies in MS and cerebral ischemia models indicate that RES may modulate immune responses, reduce oxidative damage, and limit ischemia−related neuronal injury. However, most of the current evidence derives from in vitro and animal studies, and clinical data remain limited. Moreover, the low bioavailability of RES and variability in dosing regimens represent important challenges for clinical translation. Therefore, although experimental findings support the potential neuroprotective role of RES, further well−designed clinical studies are required to determine its therapeutic relevance and safety in human neurological disorders. This narrative review was developed through a structured search of PubMed, Scopus, and Web of Science for articles published between 2000 and 2024, focusing on mechanistic, preclinical, and clinical investigations of RES in neurological disorders. This review synthesizes current evidence on the molecular and cellular mechanisms underlying the neuroprotective effects of RES, with particular emphasis on its antioxidant, anti-inflammatory, and immunomodulatory activities. By integrating findings from experimental and clinical research, the review highlights the potential of RES to modulate key pathways involved in neurodegeneration and neuroinflammation. Although further well-designed clinical studies are required to clarify its therapeutic efficacy and translational relevance, the available evidence supports continued investigation of RES as a promising candidate for neuroprotective strategies in neurological disorders.

Polymorphisms of mouse chitinase-like protein 3 (Chil3), a member of the mammalian chitinase-like protein (CLP) family, have been demonstrated to be associated with inflammatory diseases by regulating lipid metabolism. However, the specific immunomodulatory impacts of CLPs, mainly mouse CHIL3 and its human functional homologue chitinase-3-like 2 (CHI3L2), on macrophage cholesterol metabolism and atherosclerosis have remained unclear. Here, we find CLPs (CHIL3 and CHI3L2) accelerate atherogenesis in a macrophage-dependent manner. Mechanistically, we identify an autocrine mechanism through which CLPs regulate cholesterol metabolism in macrophages. Macrophage-secreted CLPs exacerbate lipid uptake by binding to CD36. CLPs exhibit glycosidase activity, targeting and hydrolyzing N-glycosylated glycans on CD36, predominantly at sites N220 and N321, thereby enhancing lipid uptake. Increased lipid influx activates mTOR in macrophages, driving their transition to a pro-inflammatory phenotype while simultaneously suppressing peroxisome proliferator-activated receptor gamma (PPARγ) expression and thus impairing ABCG1-mediated cholesterol efflux. Single-cell sequencing reveals that CLPs increase atherosclerotic foamy macrophages, favoring vascular smooth muscle cells (VSMC) transformation into foam and osteoblast-like cells. Additionally, neutralizing antibodies targeting CHI3L2 prevent and treat atherosclerosis. These findings highlight the potential of CLPs as targets for disease diagnosis and therapy.

Ferroptosis, characterized by lipid peroxidation and iron-dependent oxidative damage, is a crucial factor in various diseases. Although researchers have extensively characterized ferroptosis in cancer and neurodegenerative disorders, its interaction with pathogenic infections remains underexplored. Recent research indicates that ferroptosis contributes to host cell damage during pathogen invasions, impacting disease outcomes. This review summarizes the characteristics, mechanisms, and regulatory networks of ferroptosis. It delineates the key regulatory steps of ferroptosis during infections caused by various pathogens, including viruses, bacteria, fungi, and parasites. Additionally, it examines changes in host markers and related signaling pathways. Furthermore, this review explores the potential similarities and differences among these pathogens and discusses therapeutic strategies for addressing pathogen-related diseases through ferroptosis-dependent mechanisms.

Photothermal nanomaterials have gained significant attention in cancer treatment due to their excellent photothermal conversion properties. However, photothermal therapy alone often results in incomplete tumor ablation. To improve therapeutic efficacy, we introduce a thermo-hydrogen coupled strategy using palladium hydride (PdH) nanoparticles that combine photothermal heating with hydrogen-driven oxidative stress modulation. PdH nanoparticles were synthesized via a chemical method and systematically characterized using transmission electron microscopy (TEM), X-ray diffraction (XRD), UV-Vis spectrophotometry, and thermos response measurements. The results demonstrated that PdH nanoparticles possess small size, high structural stability, good dispersibility, and a photothermal conversion efficiency of 61.9% at 100 µg mL−1. Hydrogen release upon 532 nm laser irradiation was confirmed using methylene blue decolorization. In vitro studies demonstrated that under laser irradiation, PdH nanoparticles efficiently and stably released hydrogen, enhancing intracellular oxidative stress and leading to selective apoptosis in liver cancer cells while sparing normal liver cells. This effect resulted in an 82% cancer cell death rate, significantly surpassing that of Pd nanoparticles without hydrogen. These findings highlight the mechanistic advantage of thermo-hydrogen synergy and support PdH nanoparticles as a promising platform for controlled and selective cancer therapy.

Traumatic brain injury (TBI) disrupts central nervous system homeostasis, leading to extracellular matrix (ECM) softening and localized hypoxia and thus contributing to astrocytic activation and sustained neuroinflammation. But the interplay between ECM softening and hypoxia in regulating astrocytic activation and response remains elusive. To understand this, we developed an in vitro model incorporating tunable-stiffness hydrogels and a precise oxygen-control system to simulate the mechanical and hypoxic microenvironment of TBI. We characterized the activation-related proteins and cytokine production of astrocytes under the in vitro model, and found that soft ECM and hypoxia independently promote astrocytic activation and synergistically activate astrocytes via HIF-1α/YAP-NF-κB signaling, resulting in astrocytic redox imbalance and neuroinflammation. We further used scanning electrochemical microscopy (SECM) to track the dynamic changes in glutathione (GSH) efflux and membrane integrity in live astrocytes in situ under pathophysiological conditions of ECM softening and hypoxia. The SECM results show that the combined ECM softening and hypoxia progressively impair cellular membrane integrity and promote GSH efflux of astrocytes, corresponding to the early changes in astrocytic function and indicative of an early activation-primed state to exacerbate secondary injury of astrocytes. Last, we found that the lovastatin (a neuroprotective agent) treatment can effectively attenuate astrocytic membrane impairment and decrease GSH efflux, proving the potential of lovastatin to mitigate inflammation and preserve neuroregulatory function. Our work observes the in situ and early state changes of astrocytes under a combined mechanical-hypoxic microenvironment for the first time. The findings offer mechanistic insights into TBI pathogenesis and highlight promising strategies for early therapeutic intervention.

Spinal cord injury (SCI) triggers secondary injury cascades dominated by microglia-driven neuroinflammation, mitochondrial dysfunction, and neuronal apoptosis, leading to progressive tissue loss and persistent neurological deficits. Andrographolide, a diterpenoid lactone from Andrographis paniculata, has broad anti-inflammatory activity, but its therapeutic potential and mechanism in SCI remain unclear. Here, we combined network pharmacology with experimental validation to evaluate andrographolide after SCI. Candidate targets and pathways were screened in silico, prioritizing PI3K-Akt signaling. In a mouse contusion SCI model, andrographolide was administered intraperitoneally at 30 or 60 mg/kg starting 2 h after injury and then once daily for 14 consecutive days. PI3K-Akt pathway involvement was examined using intrathecal LY294002 under the 60 mg/kg andrographolide regimen. Behavioral assessments were conducted from 0 to 28 days post-injury to evaluate the sustained effects of early andrographolide treatment. In vitro, LPS-stimulated BV2 microglia were treated with andrographolide with or without LY294002, followed by cytokine assays, JC-1 analysis of mitochondrial membrane potential, and p-PI3K immunofluorescence. Network pharmacology identified overlapping genes between andrographolide-related targets and SCI-associated genes, and highlighted PI3K-Akt as a key enriched pathway. Consistent with this prediction, andrographolide dose-dependently restored PI3K/Akt/GSK3β/CREB phosphorylation and shifted apoptosis markers toward survival in vivo, with 60 mg/kg producing stronger effects, while LY294002 attenuated these molecular benefits. Using the selected regimen, andrographolide improved locomotor recovery and gait performance, reduced lesion cavitation, and preserved neuronal survival after SCI. In BV2 microglia, andrographolide increased p-PI3K, suppressed LPS-induced TNF-α/IL-1β/IL-6 release, and restored mitochondrial membrane potential, whereas LY294002 attenuated these effects. Collectively, andrographolide confers neuroprotection after SCI in association with activation of PI3K-Akt signaling, attenuation of inflammatory responses, preservation of mitochondrial function, and inhibition of apoptosis. Complementary BV2 cell experiments suggest that microglia-related inflammatory modulation may contribute to these protective effects.

Traumatic brain injury (TBI) is a severe central nervous system injury, but the current diagnostic methods have limitations. Identifying plasma biomarkers and developing precise and convenient detection methods are of great significance for diagnosis and prognosis prediction. This study analyzed publicly available plasma miRNA sequencing data from post-TBI samples using bioinformatics to identify potential TBI-specific miRNAs and validated them in TBI model rats. Subsequently, the study developed a biosensor based on floating gate carbon nanotubes field effect transistor (FGCNT FET) functionalized with target miRNA probes for detecting plasma miRNA concentrations and compared its performance with PCR methods. Through bioinformatics analysis, we found that plasma rno-miR-323-3p levels significantly increased after TBI, suggesting its potential as a biomarker for TBI. PCR results indicated that plasma rno-miR-323-3p concentrations were elevated in TBI rats and positively correlated with injury severity. The developed biosensor exhibited excellent electrical and sensing performance. Specifically for rno-miR-323-3p detection, the biosensor exhibited high sensitivity and specificity, good reproducibility, and a wide detection range (1 fM to 1 mu M). Compared to traditional qPCR methods, the biosensor offers the advantages of directly using plasma as the input sample, simple operation, a lower detection limit, and reduced measurement error, which indicate the significant clinical application potential.

MOF-derived catalysts have been widely investigated to enhance the catalytic activity of MgH2 due to their highly adjustable morphology and abundant active sites. While significant efforts have been made to improve the catalytic performance of these materials, the effect of particle size gradation has not been systematically investigated. Herein, a series of hierarchical Co@C-x (x = S, M, L) catalysts with controlled particle sizes were synthesized by tailoring the crystallization conditions of ZIF-67 and systematically investigated for their catalytic effects on MgH2 hydrogen storage. The results reveal that reducing the particle size of ZIF-67-derived Co@C catalysts significantly enhances hydrogen absorption and desorption kinetics. Among the samples, MgH2-Co@C-S with the smallest particle size exhibits an initial dehydrogenation temperature of 194 degrees C, which is 176 degrees C lower than that of pure MgH2, while the activation energy for desorption decreases to 93.1 kJ/mol. Moreover, it maintains 97.92 % of its hydrogen storage capacity after multiple cycles without notable degradation. This enhancement is attributed to the "hydrogen pump" effect induced by the in situ formation of Mg2Co/Mg2CoH5, nanoscale surface effect, as well as the presence of carbon species that stabilize the MgH2 matrix by preventing particle aggregation. These findings provide new insights into the size-dependent catalytic behavior of MOFderived materials and highlight the importance of hierarchically structured catalysts in optimizing MgH2 hydrogen storage performance.

Psoriasis is a chronic inflammatory skin disease, with the IL-17 pathway being a key contributor. Ym1, a positionally cloned inflammation regulatory gene linked to various disorders, has an unclear effect on skin inflammation. In this study, the role of Ym1 was investigated in both mannan and imiquimod-induced psoriasis-like dermatitis models, using Ym1-deficient congenic mice. Natural polymorphism of Ym1 influenced the development of skin inflammation, dependent on macrophages, since adoptive transferring of Ym1-deficient macrophages alleviated disease, whereas recombinant Ym1 worsened it. Particularly, Ym1 congenic mice exhibited decreased IL-17 production in innate immune cells, and depletion of γδT cells mitigated disease and lowered skin IL-17 levels. Additionally, RNA-seq analysis revealed Ym1-regulated keratinization in lesional skin. Recombination Ym1 directly influenced the inflammatory response and proliferation of mouse primary keratinocytes. Collectively, we conclude that Ym1 regulates γδT cell-derived IL-17 production and keratinocyte functionality, and thereby contributes to skin inflammation in mice.

Pancreatic cancer, recognized for its extreme lethality, is normally diagnosed at an advanced stage, leaving only a minority of patients eligible for surgical resection. Pulsed electric field (PEF) ablation, an ablative technique for clinical treatment of locally advanced pancreatic cancer, causes tumor cell death by disrupting cellular redox balance, resulting in the overproduction of reactive oxygen species (ROS) and reactive nitrogen species (RNS). In situ monitoring of the dynamic changes in ROS/RNS levels of pancreatic cancer cells under PEF ablation is important for understanding its mechanism for pancreatic cancer treatment. Here, we constructed an in vitro pancreatic cancer cell model via culturing PANC-1 cells on polyacrylamide gels with a stiffness of 4.0 kPa to simulate the mechanical microenvironment of advanced pancreatic cancer stage. We then established a theoretical model and simulated the electric field strengths of PEF ablation used for the clinical treatment of pancreatic cancer. After applying PEF ablation with these electric field strengths on the in vitro pancreatic cancer cell model, we in situ monitored the dynamic releasing processes of hydrogen peroxide (H2O2) and nitric oxide (NO) from PANC-1 cells by employing scanning electrochemical microscopy (SECM). Through quantitative analysis of SECM results, we found that the electric field strengths of PEF ablation over 1.5 kV cm-1 caused cells to exhibit periodic bursts of H2O2 and NO effluxes during the initial 5 min and then reduced to their initial values within 6 min after treatment, while mitochondrial dysfunction persisted. It demonstrates a prolonged impact of electric field strength on disrupting cellular redox balance, offering valuable insights into the mechanism of electric field strength-dependent redox imbalance of pancreatic cancer under electric ablation.

Precisely controlling the cutting of water using mechanical forces remains challenging due to water's inherent surface tension and rapid self-healing properties. Inspired by the effortless movement of water striders, a strategy is developed involving magnetic manipulation of a hydrophobic sphere across hydrophobic particle-encapsulated water (HPEW). Stable mechanical cutting of water is first demonstrated by coating its surface with hydrophobic particles (silica nanoparticles, paraffin, and polytetrafluoroethylene (PTFE)) and maintaining the water thickness below 1 mm. Through systematic theoretical and numerical analyses, it is clarified how water thickness and particle distribution influence cutting performance and accuracy. Moreover, a magnetically controlled approach is established for precise cutting, creating versatile open millifluidic chips suitable for diverse applications such as biochemical assays, chemical synthesis, and 3D cell culture. The approach thus offers a robust platform with wide-ranging implications in materials science, chemistry, physics, biomedical engineering, and microfluidics.