The mechanical properties of the tumor microenvironment serve as crucial physical cues that shape cell fate decisions. However, whether microenvironmental mechanical forces modulate ferroptosis to drive radioresistance remains unclear. Here, using PDMS-based hydrogels with tunable stiffness to establish tumor cell culture systems, we found that tumor cells cultured on stiff substrates were more sensitive to both ferroptosis and radiotherapy. Mechanistically, tumor cells grown on stiff matrices showed increased microtubule acetylation and ER sheet-to-tubule remodeling, which increased the formation of mitochondria-associated membranes (MAMs) and led to mitochondrial succinate accumulation. Succinate in turn promoted CPT1A-mediated succinylation of ACSL3, facilitating its degradation and thereby enhancing tumor cell sensitivity to ferroptosis. Collectively, these findings identify MAMs as intracellular mechanosensitive structures that regulate mitochondrial metabolism and ferroptosis in tumor cells, providing new insights into the mechanical control of ferroptosis and its implications for tumor radioresistance.

Per- and polyfluoroalkyl substances (PFASs) are a class of highly persistent synthetic chemicals that have raised significant environmental and health concerns due to their stability and bioaccumulation potential. 6:2 chlorinated polyfluoroalkyl ether sulfonic acid (6:2 Cl-PFESA), a novel alternative to traditional PFASs like perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA), has been increasingly used in various industries. However, its neurotoxic effects remain poorly understood. In this study, we compared the neurotoxic impacts of 6:2 Cl-PFESA with those of PFOS and PFOA in PC12 cells. Our findings revealed that while all three PFASs induced dose-dependent neurotoxicity, 6:2 Cl-PFESA exhibited the most potent effects, causing significantly greater reductions in cell viability and higher rates of apoptosis. Lipidomic analysis further demonstrated that high-dose exposure (100 μM) to all three chemicals profoundly altered lipid profiles, perturbing broad neurotoxicity-related networks including phosphoinositide (PI) metabolism, MAPK, and BDNF-TrkB signaling. Crucially, 6:2 Cl-PFESA uniquely induced lipidomic disruptions even at a low dose (0.025μM) reflecting human exposure levels, specifically triggering pathways such as phospholipid biosynthesis and ferroptosis. Our study underscores the urgent need to evaluate the neurotoxic risks of emerging PFAS alternatives, especially considering their widespread environmental distribution and potential to covertly impair neuronal signaling pathways.

Resina Draconis (RD), a traditional Chinese medicinal resin renowned for its potent antioxidant, anti-inflammatory, and tissue-repairing properties, holds promise for combating skin photoaging. However, its therapeutic translation has been hindered by the poor solubility and limited bioavailability of its principal bioactive constituents. In this study, we developed an RD-loaded polymeric micelle (RDPM) hydrogel to overcome these formulation challenges and evaluated its protective efficacy against UV-induced skin aging. The micellar system, composed of poloxamer surfactants and embedded within a carbomer hydrogel matrix, achieved high encapsulation efficiencies ( 80–90

To address the persistent challenges of quality consistency and efficacy stability in traditional Chinese medicine preparations arising from variations in raw materials and manufacturing processes, and the inability of conventional quality control approaches to directly link overall chemical characteristics with bioactivity, this study aimed to develop a function-oriented quality evaluation system and a new pathway for activity-relevant Q-marker screening. Using Xuezhikang capsules (XZK) as a model, high performance liquid chromatography (HPLC) fingerprints and electrochemical fingerprints were constructed for 30 batches of samples, and the Bi Value Method based on macro qualitative similarity (Sm) and macro quantitative similarity (Pm) was applied for qualitative identification and quantitative grading. A novel peak separation information energy index (PRI) was proposed to characterize chromatographic separation performance and information integrity. Antioxidant activity was evaluated using DPPH radical scavenging and Belousov-Zhabotinsky (B-Z) oscillation inhibition assays, and the relationships between fingerprint features and bioactivity were analyzed via partial least squares (PLS) modeling. The results showed that all samples exhibited high chemical similarity (Sm > 0.996), while Pm enabled classification into three quality grades and was highly correlated with total peak area. PRI demonstrated good reproducibility and complemented Pm in quality monitoring. Electrochemical fingerprints revealed more refined functional differences, further differentiating samples into five grades. PLS analysis and Pm-activity coupling consistently identified G1, G3, G4, G8, and G17 as key antioxidant-related Q-marker candidates. Overall, this integrated framework combining chromatographic profiling, electrochemical behavior, PRI evaluation, and bioactivity modeling provides a reliable, function-oriented strategy for quality grading and Q-marker discovery in traditional medicines.

BACKGROUND:Atopic dermatitis (AD) is a chronic inflammatory skin disease often exacerbated by psychological stress, yet the underlying mechanisms remain unclear. Wushe Zhiyang Pills (WZP), an oral Chinese patent medicine, is clinically used for dermatological conditions, but its efficacy and mechanisms in stress-aggravated AD have not been defined. PURPOSE:This study aims to evaluate the protective effects of WZP against AD under stress conditions, elucidate its molecular mechanisms, and identify its bioactive compounds. METHODS:A murine AD model was established using repeated DNCB application, with or without chronic restraint stress to simulate psychological stress exposure. AD-like phenotypic severity, immune dysfunction, and inflammatory responses were comprehensively assessed. Targeted oxylipin lipidomics was performed to profile lipid mediator alterations, while molecular docking, molecular dynamics simulations, and cellular thermal shift assays (CETSA) were employed to identify and validate the direct target and active components of WZP. RESULTS:WZP treatment significantly ameliorated AD-like symptoms, restored the Th1/Th2 immune balance, reduced mast cell infiltration and cutaneous inflammatory cytokine expression, and lowered serum IgE levels. Notably, psychological stress exacerbated AD by upregulating ALOX15 expression and disrupting the cutaneous oxylipin profile, with marked elevations in pro-inflammatory mediators such as 15-HETE. WZP effectively reversed these stress-induced oxylipin derangements. Mechanistically, cimifugin, a bioactive component of WZP, was identified as a direct inhibitor of ALOX15, binding stably to the enzyme and suppressing its activity, thereby restoring oxylipin homeostasis. CONCLUSION:WZP alleviates stress-aggravated AD by inhibiting ALOX15-mediated oxylipin disruption, with cimifugin as a key active component. This reveals a novel "Stress-ALOX15-oxylipin-inflammation" axis and supports WZP as a promising therapy for stress-associated AD.

Viral infections trigger cellular stress responses, and host stress proteins play key roles in antiviral defense. Here we identify stress-responsive protein arachidonate lipoxygenase-15 (ALOX15) is a critical component of mitochondrial antiviral innate immunity. Loss of Alox15 impairs mitochondrial antiviral signaling (MAVS)-mediated type I interferon production, resulting in increased susceptibility to influenza virus, an effect reversed by adeno-associated virus-mediated lung delivery of Alox15. ALOX15 translocates to mitochondria in response to H1N1 and other RNA viruses (H3N2 and human coronavirus-229E), independent of its enzymatic activity. This mitochondrial localization is also strikingly observed in peripheral blood mononuclear cells from influenza-infected individuals. Mechanistically, ALOX15 is recruited to mitochondria by polymerized MAVS, displacing the deubiquitinase USP19 and sustaining MAVS K63-linked ubiquitination and aggregation. Leveraging these insights, we developed a synergistic ALOX15 regulation-based therapeutic strategy for influenza infection by combining the ALOX15 transcriptional activator songorine with its enzymatic inhibitor PD146176. Together, our findings establish ALOX15 as an essential component of mitochondrial antiviral immunity and a promising host-directed target for antiviral therapy.

Natural Calculus Bovis (NCB) and Bovis Calculus Sativus (BCS) are both used clinically but exhibit different therapeutic effects, the material basis of which remains unclear. Lipid-soluble components are hypothesized to contribute to these differences, however the compositional variations at the lipid level have not been fully characterized. In this study, untargeted UHPLC-MS/MS lipidomics was employed to compare the lipid profiles of NCB and BCS. The results indicated that both samples contained fatty acyls, glycerolipids (GL), glycerophospholipids (GP), sphingolipids (SP), and sterol lipids (ST), indicating a conserved core lipid framework. However, NCB showed higher lipid complexity and abundance. NCB contained higher levels and greater molecular diversity of intact GP (phosphatidylcholines and phosphatidylethanolamines), SP (ceramides), fatty acid-derived lipids (polyunsaturated N-acylethanolamines, PU-NAE), and sterol esters. In comparison, BCS exhibited a simplified profile dominated by GL, with reduced GP, SP and ST species, enrichment of lysophospholipids and bile acid-related sterols, and lower overall lipid abundance. These differences may result from distinct formation processes: prolonged in vivo maturation for NCB versus time-limited in vitro culturing for BCS. This study expands the current understanding of the material basis of NCB and BCS, from a lipidomic perspective, offering molecular insights into its pharmacological properties and providing a scientific foundation for the material basis, quality control, standardization and rational clinical application of Calculus Bovis.

ETHNOPHARMACOLOGICAL RELEVANCE:Diabetic kidney disease (DKD) is a major diabetic complication with limited therapeutic options. Vaccinium myrtillus L. (bilberry) has a long history in European traditional medicine for treating microvascular disorders, inflammatory conditions, and urinary system ailments. As a traditional medicinal food rich in water-soluble anthocyanins, it has been reported to exert nephroprotective effects; however, its bioactive constituents and molecular mechanisms remain poorly defined. AIM OF THE STUDY:This study aimed to investigate the renoprotective effects of a water-soluble anthocyanin-rich bilberry extract (VmE) in DKD, to identify its bioactive components, and to elucidate the underlying molecular mechanisms. MATERIALS AND METHODS:A high-fat diet/streptozotocin-induced DKD mouse model was employed. The chemical profile of VmE was characterized by LC-MS. Renal function, histopathology, and metabolomics were evaluated. Integrated network pharmacology, molecular docking, microscale thermophoresis (MST), and molecular dynamics (MD) simulations were conducted to identify and validate key molecular targets. RESULTS:VmE dose-dependently improved renal function, ameliorated tubulointerstitial injury, and restored metabolic homeostasis. Delphinidin-3-O-glucoside (Dp3Glc) and cyanidin-3-O-glucoside (Cy3Glc) were identified as the major anthocyanins. Both MST and MD simulations confirmed that Dp3Glc and Cy3Glc directly bind to arachidonate 15-lipoxygenase (ALOX15). This interaction inhibited ALOX15-mediated lipid peroxidation, resulting in reduced malondialdehyde (MDA) accumulation and iron overload, as well as restored glutathione (GSH) homeostasis, thereby attenuating ferroptosis in renal tubular cells. CONCLUSIONS:VmE through its major anthocyanins Dp3Glc and Cy3Glc, alleviates DKD by directly targeting ALOX15 and suppressing lipid peroxidation-driven ferroptosis. These findings establish a pharmacological basis for developing VmE as a standardized nutraceutical for DKD management, bridging traditional use with modern mechanistic evidence.

Conventional quality control protocols for foods and medicinal products seldom integrate chemical fingerprints with bioactivity readouts, limiting their discriminative power for complex matrices. We developed a digitalization workflow that employs full-spectrum temporal-integration analysis (FTIA) and three-dimensional peak-max projection method (3D-PMPM) on HPLC-DAD data for big red robe tea. The 200-400 nm spectral cube is compressed by FTIA and 3D-PMPM and then evaluated against a reference using a binary evaluation system (BES; Sm, Pm). A post-column on-line ABTS assay produced time-aligned antioxidant fingerprints. A six-parameter "5 + 2" bracketing check verified sequence stability. Median-based activity maps located robust hotspots at peaks P2-P3 and P12; an abundance-weighted Antioxidant Activity Index provided consistent lot ranking. Process screening showed that mild citric acid (0.60-0.85%) with a 1:50 solid-liquid ratio and 20-30 min ultrasonication maximized the digital fingerprints. The workflow delivers quality control-ready, auditable metrics that co-register spectral fingerprints, composition, and antioxidant activity, and is transferable to other foods.

Cancer-associated fibroblasts (CAFs) are major stromal components of the tumor microenvironment, yet how their metabolic states shift during therapy and influence anti-tumor immunity remains unclear. By integrating clinical cancer samples, single-cell RNA analyses, and functional studies, we identify a chemotherapy-conditioned PTGER3+ CAF subset characterized by enhanced lipid oxidation. This metabolic reprogramming strengthens antitumor immunity by promoting CD8+ T cell activation and cytotoxicity through the suppression of PTEN-related signaling. Clinically, higher proportions of therapy-induced PTGER3+ CAFs correlate with improved treatment responses and better patient prognosis. Together, these findings reveal a previously unrecognized stromal metabolic adaptation that supports CD8+ T cell immunity and highlight CAF-driven lipid oxidation and its regulation of CD8+ T cell PTEN signaling as potential avenues to enhance chemotherapy and immunotherapy efficacy.

Skin aging is characterized by reduced hydration, elasticity loss, wrinkles, and impaired skin function. Hyaluronidase-mediated hyaluronic acid degradation, together with oxidative stress, inflammation, and fibroblast senescence, is a key driver of this process. Therefore, safe hyaluronidase inhibitors with additional bioactivities may offer potential for skin-aging intervention. In this study, royal jelly protein hydrolysates (RJPH) were generated by enzymatic hydrolysis and characterized using UHPLC-MS/MS-based peptidomics. Potential hyaluronidase-inhibitory peptides were screened through virtual screening, molecular docking, and in vitro enzyme inhibition assays. A novel hyaluronidase-inhibitory peptide, with the amino acid sequence abbreviated as LVSAFK, was further evaluated using molecular dynamics simulation, the oxygen radical absorbance capacity (ORAC) assay, and cell-based assays in human skin fibroblastsusing lipopolysaccharide (LPS)-induced inflammation and H2O2-induced senescence models. LVSAFK displayed hyaluronidase inhibitory activity with an IC50 of approximately 301.6 mu M. Docking and molecular dynamics analyses indicated stable binding mediated by hydrogen bonds, salt bridges, and cation-It interactions. In addition, LVSAFK exhibited peroxyl radical-scavenging activity, suppressed LPS-induced IL-1 beta production, and reduced SA-beta-galactosidase-positive senescent fibroblasts under H2O2 stress. Collectively, these findings identify LVSAFK as a multifunctional royal jelly-derived peptide with in vitro activities relevant to skin aging, warranting further mechanistic and in vivo studies for potential anti-skin-aging applications.

Systemic rotavirus (RV) infection poses a substantial health challenge in neonates, but the underlying pathogenesis remains elusive. In RV-infected neonatal mice and infants with biliary atresia (BA), we discovered that persistent type I interferon (IFN-I) signaling upregulated hepcidin expression in hepatocytes and TREM2+ macrophages. This impaired SLC40A1-mediated iron excretion, leading to lipid peroxidation- and ferroptosis-mediated tissue damage. In mice deficient in Slc40a1 in myeloid cells, iron accumulation promoted RV replication and IFN-I activation in Kupffer cells. Blocking IFN-I-hepcidin signaling and iron chelation reduced RV-induced tissue damage in mice. Folic acid suppressed IFN-I-hepcidin-iron signaling in mice, and in an open-label clinical trial, folic acid supplementation in infants with BA reduced cholangitis and liver transplantation rates. Our data show that hepcidin-iron dysregulation plays a critical role in neonatal RV infection and reveal therapeutic targets for BA and other RV-related neonatal diseases. The clinical trial was registered in the Chinese Clinical Trial Registry ChiCTR2100050992.

Radix Fici Simplicissimae (RFS) is a commonly consumed edible plant in southern China, renowned for its distinctive aroma and nutritional value. However, its complex chemical composition and the variations in growth environments result in significant differences in quality across the market. Existing identification methods are inadequate for effective quality control. Therefore, a comprehensive and multi-dimensional approach is required to ensure consistent quality, food safety, and customer satisfaction. In this study, we employed high-resolution mass spectrometry to identify the major chemical constituents of RFS. Additionally, we established fingerprints for 26 batches of samples using high-performance liquid chromatography (HPLC) with a diode array detector (DAD), ultraviolet (UV) spectroscopy, and Fourier-transform infrared (FT-IR) spectroscopy. These fingerprints were then analyzed using the Systematic Quantitative Fingerprinting Method (SQFM) to assess the quality of the samples. Antioxidant activity was evaluated, and the relationship between HPLC fingerprint peaks and this activity was explored through partial least squares (PLS) modeling. The Pearson coefficient method was applied to assess the correlation between UV fingerprints and antioxidant activity. Our findings suggest that the PLS model can effectively predict antioxidant components, while UV fingerprints help identify the active structural features of antioxidants. Moreover, mass spectrometry imaging (MSI) revealed that the principal bioactive components of RFS are predominantly localized near the xylem. In conclusion, this study presents a multi-faceted approach to analyzing and evaluating the quality of RFS, which can be applied to quality control in both plant and food industries. This framework also lays the groundwork for further research into the bioactive substances in RFS, their spatial distribution, and their potential application in the quality control of other complex plant and food samples.

Redox‐based diagnostic and therapeutic applications have long suffered from a shortage of suitable drugs and probes of high specificity. In the context of anti‐ferroptosis research for neurological diseases, the inaccessibility of a blood‐brain barrier permeable (BBB) small molecular ferroptosis inhibitor, and the lack of specific ferroptosis probes seriously impeded a deeper understanding of the mechanism of ferroptosis and the development of clinically applicable drugs. We here report a novel 1,3,4‑thiadiazole‐functionalized drug‐like ferrostatin analogue entitled Ferfluor‐1 with superior anti‐ferroptosis potency, favorable BBB permeability and in vivo activity against stroke and Parkinson’s disease. Moreover, the exclusive pseudo excited‐state intramolecular proton‐transfer (ESIPT) property of Ferfluor‐1 via a long‐distance hydrogen‐bonding network facilitated it as the first sensitive ratiometric photoluminescent probe to detect phospholipid hydroperoxides and a specific indicator for the fluctuation of ferroptosis. These unprecedented advantages not only engendered Ferfluor‐1 as a potential tool for ferroptosis‐related diagnostic and therapeutic applications in the central nervous system, but also paved the way to developing new theragnostic agents for precision redox detection and regulation.

Plant metabolomes are highly complex due to their extensive structural diversity,making their identification challenging with-out comprehensive reference databases.Liquid chromatography-mass spectrometry(LC-MS)is commonly utilized for plant meta-bolome analysis,capable of generating numerous informative MS/MS spectra[1-3].However,only a small portion of these spec-tra can be effectively annotated[4].

Despite the known benefits of tea (Camellia sinensis), the functional potential of tea-derived proteins remains largely unexplored. To investigate this, tea proteins were extracted and enzymatically hydrolyzed with alkaline protease to obtain tea protein hydrolysates (TPH). Characterized by ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS), TPH were screened via integrated in silico and molecular docking for dual-action peptides targeting tyrosinase inhibition and antioxidant activity. Two peptides, EGFG and FGDPHG, were identified and validated as potential tyrosinase inhibitors and antioxidants, exhibiting strong free radical scavenging and iron-chelating capabilities in cell-free systems. In B16-F10 melanoma cells, both peptides significantly reduced α-melanocyte-stimulating hormone (α-MSH)-induced melanogenesis and protected against hydrogen peroxide (H2O2)-induced oxidative stress. Their whitening efficacy was further confirmed in a zebrafish model. These findings establish EGFG and FGDPHG as novel, dual-action whitening peptides, highlighting their potential as functional ingredients in cosmetics and nutraceuticals.

Despite wide variation, each cell type has an optimal size. Maintaining optimal size is essential for cellular fitness and function but the biological basis for this remains elusive. Here, we performed fitness analysis involving genome-wide CRISPR-Cas9 knockout data from tens of human cell lines and identified that cell size influences the essentiality of genes related to mitochondria and membrane repair. These genes also included glutathione peroxidase 4 (GPX4), which safeguards membranes from oxidative damage and prevents ferroptosis-iron-dependent death. Growth beyond normal size, with or without cell-cycle arrest, increased lipid peroxidation, resulting in a ferroptosis-sensitive state. Proteomic analysis revealed cell-cycle-independent superscaling of endoplasmic reticulum, accumulation of iron, and lipidome remodeling. Even slight increases from normal cell size sensitized proliferating cells to ferroptosis as evidenced by deep-learning-based single-cell analysis. Thus, lipid peroxidation may be a fitness trade-off that constrains cell enlargement and contributes to the establishment of an optimal cell size.

Ganoderma lucidum (G. lucidum), has been documented as a medicinal herb in classical texts and officially recognized in both Eastern and Western pharmacopeias. G. lucidum spore oil (GLSO), a lipid substance extracted from sporoderm-broken spores, has shown potential in enhancing immune function and prolonging the survival of tumor patients. However, the mechanisms underlying GLSO’s immunomodulatory effects remain poorly unknown. The effect of psychological stress on tumor progression and macrophage phagocytosis was analyzed by an in vivo small animal imaging system and flow cytometry. The effect of psychological stress on phospholipid composition in mice was investigated by LC–MS/MS based lipidomic analysis. The effectiveness of GLSO in tumor-bearing mice subjected to restraint stress was observed by tumor burden and phagocytosis of macrophages. Finally, the underlying mechanism of GLSO on macrophage phagocytosis in mice subjected to psychological stress was explored by RNA-seq, and the FcγR/SYK-mediated macrophage phagocytosis pathway was confirmed by qPCR, Western blotting, and confocal laser technology. Our study discovered that psychological stress-triggered tumor progression is contributed to by liposoluble components-impaired macrophage phagocytosis. Lipidomics analysis further identified lysophosphatidylinositol [LPI (18:0)] as a key factor suppressing macrophage phagocytic capacity under psychological stress. GLSO was shown to mitigate psychological stress-evoked tumor progression by enhancing macrophage-mediated phagocytosis of tumor cells in vivo. Mechanistically, transcriptomics analysis revealed that the LPI-mediated FcγR phagocytosis pathway is a crucial axis driving the therapeutic effect of GLSO under psychological stress. Our findings illustrate that psychological stress-promoted cancer progression is contributed by the critical liposoluble components LPI (18:0)-mediated FcγR phagocytosis signaling inhibition. GLSO alleviates the dampened phagocytosis of macrophages caused by stress through regulating LPI/FcγR-mediated phagocytosis-related pathways, underscoring its potential as a therapeutic intervention for stress-related tumor progression.