Search Results (5,565)

Benzo(a)pyrene (B[a]P) is a pervasive freshwater pollutant, yet its toxicity to the fish gallbladder remains poorly understood. This study investigated the toxicological impacts of 2.5 and 25 μg/L B[a]P on common carp (Cyprinus carpio) using histological, transcriptomic, and single-cell RNA sequencing (scRNA-seq) analyses. Results showed that the gallbladder is a primary site for B[a]P accumulation. High B[a]P concentrations caused vacuolar degeneration of mucosal epithelial cells and nuclear deformities. Transcriptomic analysis revealed that B[a]P stress triggered autoimmune homeostasis imbalance and overinhibited apoptosis. scRNA-seq identified cellular heterogeneity changes, specifically T-cell impairment and epithelial cell (EC) proliferation. Mechanistically, T-cell reduction was linked to the T-cell 2 subset, while EC proliferation involved EC 0 and EC 4 subsets, all participating in the apoptosis pathway. These findings demonstrate that the apoptosis pathway is a key target of B[a]P toxicity in the gallbladder. This work provides a cellular-level framework for assessing environmental polycyclic aromatic hydrocarbon (PAH) risks in aquaculture. Full article

Carbosulfan is a widely used carbamate insecticide, yet its mechanisms of respiratory toxicity remain poorly understood. This study integrated network toxicology, untargeted metabolomics, and molecular docking to systematically investigate the potential mechanisms of carbosulfan-induced respiratory toxicity in male Sprague Dawley rats. Rats were administered a single oral dose of carbosulfan (125 or 250 mg/kg) and assessed after 12 h. Exposure resulted in significant pathological lung damage, characterized by disrupted alveolar architecture, inflammatory cell infiltration, and increased serum levels of the pro-inflammatory cytokines IL-6, IL-1β, and TNF-α. Network toxicology analysis identified 51 potential targets associated with respiratory toxicity, with core targets including SRC, EGFR, PTGS2, CXCL8, CYP3A4, and NR3C1. Enriched pathways were primarily related to neuroactive ligand–receptor interaction, VEGF signaling, and arachidonic acid metabolism. Untargeted metabolomics revealed significant metabolic perturbations in pathways central to antioxidant defense and energy homeostasis, including glutathione metabolism, the tricarboxylic acid cycle, and arginine biosynthesis. Molecular docking confirmed stable in silico binding affinities between carbosulfan and the predicted core targets. Integrative analysis suggests that carbosulfan exposure is associated with respiratory damage, potentially through interconnected mechanisms involving oxidative stress, inflammation, and disruption of cell signaling and metabolic enzyme systems. However, given the acute high-dose nature of the model and the interpretative integration of multi-omics data, these findings should be considered hypothesis-generating. This study provides a novel system-level perspective on carbosulfan-induced respiratory toxicity and highlights key pathways and targets for future validation in chronic exposure models. Full article

Oral squamous cell carcinoma (OSCC) remains a major cause of cancer-related mortality worldwide, with limited biomarker-driven tools for risk stratification. CD147 is a membrane glycoprotein implicated in tumor metabolism, invasion, immune evasion, and therapy resistance. This study aimed to evaluate the prognostic and predictive relevance of CD147 expression in distinct tumor compartments of OSCC. Formalin-fixed tumor samples from 229 OSCC patients were analyzed via tissue microarray and immunohistochemistry to assess CD147 expression in the tumor center, periphery, and adjacent mucosa. Associations with clinicopathological parameters, survival, and therapy response were evaluated using non-parametric statistical tests, Kaplan–Meier, multivariate Cox, and binary logistic regression analyses. Complementary transcriptomic and immunological analyses were performed using The Cancer Genome Atlas (TCGA), the University of Alabama at Birmingham Cancer data analysis (UALCAN), Tumor and Immune System Interaction Database (TISIDB), and the Genotype-Tissue Expression (GTEx) project’s datasets. Low CD147 expression in the tumor invasive front was independently associated with improved overall survival, while expression in the tumor center or mucosa showed no prognostic value. No significant associations between CD147 and adjuvant therapy response were identified. TCGA-based analyses confirmed CD147 overexpression in tumors and its correlation with immunosuppressive signaling and resistance-associated transcriptional networks. Peripheral CD147 expression serves as a compartment-specific, independent prognostic marker in OSCC in this retrospective single-center cohort. Its spatially restricted prognostic relevance and association with immune modulation and therapy resistance highlight CD147 as a promising candidate for future biomarker-driven and therapeutic strategies. Full article

Neonicotinoid insecticides and fluoroquinolone antibiotics frequently co-occur in aquatic and terrestrial environments, posing a threat to human health, yet their combined neurotoxic potential remains poorly characterized. This study aimed to assess the cytotoxicity of typical neonicotinoids and fluoroquinolones as well as their mixtures in human neuroblastoma SK-N-SH cells and identify affected pathways. SK-N-SH cells were exposed to clothianidin (CLO), imidacloprid (IMI), enrofloxacin (ENR), and ofloxacin (OFX) individually and in fixed-ratio mixtures (50% of each compound’s IC₅₀) for 24 h and 48 h, and cell viability was quantified using the alamarBlue^® method. Single-compound dose–response testing showed time-dependent cytotoxicity, with higher potency for fluoroquinolones (24 h IC₅₀: ENR 1.446 mM, OFX 2.742 mM; 48 h IC₅₀: ENR 0.826 mM, OFX 2.005 mM) than neonicotinoids (24 h IC₅₀: IMI 4.754 mM, CLO 5.356 mM; 48 h IC₅₀: IMI 3.631 mM, CLO 4.029 mM). Concentration-addition analysis indicated that most mixtures produced synergistic interaction in reduction in cell viability, with ENR+OFX showing the strongest effect at 48 h (Observed viability 7.138% vs. Predicated viability 82.368%). RNA-seq (24 h) revealed that binary mixtures generally induced more differentially expressed genes than single exposures, and ENR-containing mixtures showed the largest transcriptomic shifts, enriching pathways related to cellular stress and injury as well as neuronal signaling and connectivity. RT-qPCR validated the changes in expressions of five key neurobiology-relevant genes (LMO3, NOS1, ADCY8, FGF7 and TNFRSF12A). These findings highlight the importance of assessing insecticide–antibiotic mixtures when evaluating their hazards in environment. Full article

Background: Overcoming therapeutic resistance and tumor microenvironment-mediated survival remains a major challenge in ovarian cancer treatment. This study investigated the individual and combined antitumor effects of etoposide and curcumin in human ovarian cancer models, with emphasis on synergistic interactions, apoptosis induction, tumor microenvironment modulation, and oxidative stress-associated mechanisms. We hypothesized that the combination of etoposide and curcumin exerts enhanced antitumor activity through an integrated mechanism involving DNA damage-associated apoptotic signaling, tumor microenvironment modulation, and ROS-associated cellular stress, rather than through a single dominant pathway. Methods: Cell viability was assessed using the MTT assay, and drug–drug interactions were quantitatively evaluated using the Chou–Talalay and Bliss independence models. Apoptosis was analyzed by Annexin V/propidium iodide flow cytometry, caspase-8/9 activity assays, and cell cycle analysis. Transcriptional regulation of apoptosis- and microenvironment-related genes was examined by quantitative real-time PCR. Inflammatory and angiogenic cytokines were measured by ELISA. Therapeutic efficacy was further validated in three-dimensional (3D) tumor spheroid models using morphological assessment and adenosine triphosphate (ATP)-based viability assays. The contribution of reactive oxygen species (ROS) was evaluated using antioxidant pretreatment. Results: The etoposide–curcumin combination demonstrated significantly enhanced antiproliferative and pro-apoptotic effects compared with either agent alone, with consistent synergism observed across quantitative interaction models. Combination treatment increased apoptotic cell death, activated both intrinsic and extrinsic apoptotic pathways, and induced G2/M cell cycle arrest. In parallel, inflammatory and angiogenic signaling was markedly suppressed at both transcriptional and protein levels. These effects were preserved and amplified in 3D tumor spheroid models, where combination therapy induced pronounced spheroid shrinkage and reduced viability. Antioxidant pretreatment partially attenuated ROS generation and cytotoxicity, indicating that oxidative stress contributes to, but does not fully account for, the observed antitumor effects. Conclusions: The combination of etoposide and curcumin exerts synergistic and multi-layered antitumor effects in ovarian cancer models by integrating apoptosis induction, tumor microenvironment modulation, and ROS-associated mechanisms. These findings support further preclinical evaluation of this combination as a rational therapeutic strategy for ovarian cancer. Full article

DENSE AND ERECT PANICLE 2 (DEP2) is a key pleiotropic gene regulating panicle architecture and grain shape in rice. To explore its favorable allelic variations, a stably inherited mutant with short, round and thick grains (srt) was identified in this study using the Japonica rice variety ‘Zhonghua 11’ (ZH11) as the genetic background. Compared with ZH11, srt exhibited reduced plant height and a shorter panicle length but significantly increased numbers of primary branches, secondary branches, and grains per panicle. The mutant grains showed decreased length, increased width, and thickness, ultimately leading to reductions in both thousand-grain weight and grain yield per plant. Cytological analysis revealed that both the longitudinal and transverse dimensions of hull cells in srt were significantly enlarged, while the number of longitudinal cells per unit length decreased. This suggests that srt remodels grain shape by altering the balance between cell division and expansion. Genetic analysis indicated that the phenotype is controlled by a single recessive gene. Map-based cloning and whole-genome resequencing localized the target gene to chromosome 7 and confirmed that srt is the DEP2 gene. Sequence analysis revealed the presence of a large fragment insertion of approximately 87 kb entirely within the seventh exon region of the DEP2 gene in srt. However, unlike most reported DEP2 mutants, in which the numbers of primary branches, secondary branches, and grains per panicle showed no significant difference from the wild type, the srt mutant exhibited a significant increase in all these traits. This unique phenotypic combination expands the understanding of the functional diversity of this gene. This study provides new genetic material for further elucidating the molecular mechanisms by which DEP2 regulates panicle development and for molecular design breeding of rice grain shape. Full article

Herein, we present a comprehensive single-cell investigation of the biochemical and metabolic responses of normal human colon fibroblasts (CCD-18Co) and colorectal adenocarcinoma cells (Caco-2) to supplementation with the amino acids leucine, threonine, and arginine, employing State-of-the-Art Raman spectroscopy and Raman imaging. This fully label-free and noninvasive methodology enabled high-spatial-resolution mapping of intracellular components, providing unprecedented insight into subcellular biochemical organization and metabolic remodeling associated with colorectal carcinogenesis. By synergistically integrating Raman spectroscopic data with advanced chemometric methods, we demonstrate robust, reproducible discrimination between normal and malignant colon cells, both in their native state and after amino acid treatment, based solely on their intrinsic vibrational fingerprints. Partial Least Squares Discriminant Analysis (PLS-DA) and one-way ANOVA revealed that perturbations in lipid metabolism and protein composition constitute key molecular determinants underlying the observed phenotypic divergence between control and amino acid–supplemented cells. Notably, detailed analysis of diagnostic Raman band intensity ratios (2845/3015, 2845/2930, 3015/2888, and 1444/1256) uncovered pronounced amino acid–driven alterations in metabolic pathways at the single-cell level. Raman imaging further enabled spatially resolved visualization of these biochemical shifts and changes in Raman band intensities, highlighting distinct lipid- and protein-rich subcellular domains that respond differentially to amino acid exposure in normal versus cancerous cells. Collectively, our findings establish Raman spectroscopy combined with chemometric analysis as a powerful and sensitive platform for decoding amino acid–induced metabolic reprogramming in colorectal cells. This approach deepens the mechanistic understanding of nutrient–cancer cell interactions and opens new avenues for the development of Raman-based strategies in cancer diagnostics and therapeutic response assessment. Full article

Environmental pollutants are persistent chemicals that pose substantial risks to human health, contributing to global mortality and economic burden. In real-world situations, exposure rarely occurs to single compounds; instead, people are chronically exposed to complex mixtures at low concentrations. However, most regulatory frameworks still rely on single-substance risk assessments, potentially underestimating the hazards associated with combined exposures. This study investigated the cytotoxic interactions of binary mixtures of five environmentally relevant pollutants: bisphenol A (BPA), bisphenol A diglycidyl ether (BADGE), dibutyl phthalate (DBP), di(2-ethylhexyl) phthalate (DEHP), and perfluorooctanoic acid (PFOA), using the human lymphoblast cell line NALM-6. Cells were exposed for 72 h to each compound individually and to all possible binary combinations, reflecting concentrations reported in human plasma or serum. Cell viability was assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, and interactions were analyzed using the Bliss model of independence and two-way analysis of variance (ANOVA). Intracellular reactive oxygen species were measured using the 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA) probe to explore the involvement of oxidative stress. Synergistic interactions were observed under specific conditions, although not all statistically identified interactions corresponded to biologically significant effects. The BPA-DBP combination produced the highest cytotoxicity when both pollutants were present at 100 nM (31%), consistent with a strong synergistic effect. A similar pattern was observed for BADGE-BPA. ROS production was partially associated with cytotoxicity in these selected mixtures. Overall, these findings highlight the importance of distinguishing statistical synergy from toxicological relevance. Full article

Fate-mapping studies have challenged the longstanding view of the adult mammalian heart as a post-mitotic organ, suggesting limited cardiomyocyte renewal. This has spurred efforts to determine whether selected cardiac stromal cells have regenerative potential; however, their contribution to cardiac regeneration has been found to be minimal compared with that of cardiomyocyte proliferation. Despite this, transplantation of some cardiac stromal cell populations has shown therapeutic potential through paracrine signalling. The identity of the paracrine-active stromal cell populations remains unclear due to overlapping characteristics with other cardiac stromal cell populations, such as fibroblasts, mesenchymal cells, and pericytes. This study sought to clarify the transcriptional identity and heterogeneity of adult mouse cardiac stromal cells by developing a cardiac collagenase–trypsin protocol and comparing it to the established method for isolating cardiosphere-derived cells (CDCs). This novel protocol resulted in a higher cell yield and shorter expansion time, and the resulting cells showed superior survival under serum starvation compared to commercially acquired cardiac fibroblasts (CFs). Single-cell qRT-PCR analysis revealed that collagenase–trypsin cells (CTs) and CDCs share similar gene expression profiles, distinct from those of CFs. Notably, CTs exhibited higher expression of Tcf21 and lower expression of Tbx5, suggesting an epicardial-derived fibroblast phenotype, whereas Tbx5 was enriched in CDCs and CFs, reflecting heterogeneity within the cardiac fibroblast compartment. This study offers insights into the complex identity of cardiac stromal cells and concludes that CTs closely resemble CDCs but can be generated more rapidly, making them a robust and efficient source of paracrine-active cardiac stromal cells. Full article

Phospholipases A₂ (PLA₂s) are key mediators of the cytotoxic and inflammatory activities of snake venoms. While PLA₂ isoforms from Bothrops diporus venom have been characterized and shown to possess antimetastatic and antiangiogenic properties, their impact on mitochondrial bioenergetics and immune modulation has not yet been investigated. In this study, we examined the bioenergetic and immunomodulatory effects of B. diporus PLA₂s using integrated biochemical, metabolic, and multiplex cytokine analyses. In MDA-MB-231 breast cancer cells, pooled PLA₂s induced a dose-dependent decrease in MTT-reducing activity, increased mitochondrial ROS, caused Δψm hyperpolarization, and decreased NADH autofluorescence, collectively indicating sustained mitochondrial stress. Real-time impedance measurements further revealed a marked inhibition of cell proliferation. In human PBMCs, pooled PLA₂s elicited a dynamic cytokine program, inducing early cytotoxic (Granzyme B) and chemotactic (CCL2, CCL3, CCL4) mediators, followed by late pro-inflammatory and regulatory factors such as IL-6, TNF-β, IL-10 and IL-15. Analysis of a single purified PLA₂ isoform (Fraction 6) confirmed activation of the canonical IL-6/TNF-α/IL-1β axis but uniquely induced IL-10 and CCL20, revealing isoform-specific immunomodulatory properties. Altogether, these findings provide the first integrated characterization of mitochondrial and immune perturbations induced by B. diporus PLA₂s, expanding their recognized biological scope and underscoring their potential as molecular templates for novel pharmacological strategies targeting mitochondrial vulnerabilities or modulating the tumor immune microenvironment. Full article

Bee pollen is a nutrient-dense food; however, its dense cell wall limits the bioavailability and digestive absorption of nutrients. This study established a co-fermentation process that combines Lactobacillus strains isolated from bee bread with commercial probiotics to improve the nutritional profile and functional properties of bee pollen. L. acidophilus (LBA1) and L. plantarum (LBP3) were isolated from bee bread and used for single-strain fermentation of bee pollen and its co-fermentation with commercial probiotics. The results indicated that fermentation increased the protein, free amino acid, vitamin C, and flavonoid contents. The co-fermentation product (FHL-99) of LBP3 and the commercial inoculant (99 strains) exhibited the highest cell wall disruption rate (67.57%) in artificial intestinal juice. Ex vivo activity analysis revealed enhanced DPPH, hydroxyl, and ABTS⁺ radical scavenging capacities of fermented bee pollen. Its inhibitory effects on hyaluronidase activity and protein thermal denaturation were also enhanced. FHL-99 demonstrated optimal performance across multiple indices, achieving a DPPH radical scavenging rate of 77.46% and hyaluronidase inhibition rate of 37.38%. In conclusion, synergistic co-fermentation can disrupt pollen cell walls and enrich bioactive constituents, providing an efficient biotechnological approach for the development of high-quality fermented bee pollen products. Full article

Root rot disease severely impacts the yield of Cardamine violifolia, a selenium-enriched cruciferous vegetable. However, the causal pathogens and effective control strategies of this disease remain poorly characterized. This study systematically isolated and identified three key pathogens from diseased tissues in the Enshi region: Aspergillus costaricensis, Mucor circinelloides cf. lusitanicus, and Fusarium pernambucanum. Morphological characterization, phylogenetic analysis, and pathogenicity testing were conducted. Candidate fungicides were screened using plate inhibition assays, and combinations were optimized and validated through soil drenching experiments. While propiconazole showed broad-spectrum activity, its efficacy against Aspergillus and Mucor was suboptimal. A novel ternary compound fungicide, T10, combining propiconazole, hymexazol, and difenoconazole, demonstrated significantly enhanced potency with EC₅₀ values of 7.313, 12.2983, and 0.1781 mg/L against the three pathogens, representing reductions of 66.0%, 77.7%, and 92.1% compared to the most effective single application of propiconazole. At 10 mg/L, T10 increased inhibition rates by 62.62%, 77.53%, and 20.85% against the three pathogens, respectively, compared to propiconazole alone. Propidium iodide (PI) staining revealed increased cell membrane permeability in T10-treated pathogens, suggesting that membrane damage may contribute to its antifungal effect. This study provides a robust scientific basis for sustainable disease management of this high-value selenium-enriched vegetable crop. Full article

Tumor-associated macrophages (TAMs) are pivotal in the clear cell renal cell carcinoma (ccRCC) microenvironment. Methylenetetrahydrofolate dehydrogenase 2 (MTHFD2), a central enzyme in one-carbon metabolism, is increasingly recognized for its oncogenic roles in both cancer cells and immune compartments. We integrated bulk and single-cell transcriptomic datasets to interrogate the expression, prognostic impact, and immunomodulatory landscape of MTHFD2 in ccRCC. Robust differential expression, meta-analysis, Cox regression, and cell type deconvolution were performed. MTHFD2 expression and its association with prognosis were validated using tissue microarrays (TMAs), multiplex IHC, and in vitro macrophage polarization assays. MTHFD2 was upregulated in ccRCC tumors and associated with poor prognosis across multiple cohorts. High MTHFD2 expression remained an independent prognostic marker after adjustment for clinical stage. Single-cell analyses identified macrophages as the principal immune subpopulation expressing MTHFD2, with MTHFD2+ macrophages displaying a transcriptional signature of immunosuppression and metabolic adaptation. In vitro, MTHFD2-induced M2 macrophage polarization was reversed by DS18561882, promoting M1 polarization. MTHFD2 is a robust biomarker for poor prognosis in ccRCC, influencing tumor–immune interactions and macrophage polarization. Targeting MTHFD2 may represent a dual-action strategy to suppress tumor growth and reprogram the tumor immune microenvironment. Full article

Fibrosis is a hallmark of the tumor microenvironment in many solid cancers, driving tumor progression, immune evasion, and treatment resistance; however, the molecular and cellular mechanisms underlying fibrogenesis—particularly stromal–immune crosstalk across organs—remain incompletely understood, compounded by organ-specific heterogeneity and a lack of reliable immune-related biomarkers. To address this, we performed an integrative single-cell RNA sequencing (scRNA-seq) analysis of fibrotic tissues from four major organs—liver, lung, heart, and kidney—alongside non-fibrotic controls, applying unsupervised clustering, trajectory inference, cell–cell communication modeling, and gene set variation analysis (GSVA) to map the fibro-immune landscape. Our analysis revealed both conserved and organ-specific features: fibroblasts were the dominant extracellular matrix (ECM)-producing cells in liver and lung, whereas endothelial-derived stromal populations prevailed in heart and kidney. Immune profiling uncovered distinct infiltration patterns—macrophages displayed organ-specific polarization states; T cells were enriched for tissue-resident subsets in lung and mucosal-associated invariant T (MAIT) cells in liver; and B cells exhibited marked heterogeneity, including a pathogenic interferon-responsive subset prominent in pulmonary fibrosis. GSVA further identified divergent signaling programs across organs and lineages, including TGF-β/TNF-α in the heart, NOTCH/mTOR in the kidney, glycolysis/ROS in the lung, and KRAS/interferon pathways in the liver. Cell–cell communication analysis highlighted robust crosstalk between macrophages, T/B cells, and stromal cells mediated by collagen, laminin, and CXCL signaling axes. Together, this cross-organ atlas delineates a highly heterogeneous fibro-immune ecosystem in human fibrotic diseases, revealing shared mechanisms alongside organ-specific regulatory networks, with immediate translational implications for precision anti-fibrotic therapy, immunomodulatory drug repurposing, and the development of context-specific biomarkers for clinical stratification and therapeutic monitoring. Full article

The involvement of the immune system in pulmonary fibrosis is established, the precise contributions of tissue-resident memory T (T_RM) cells are still poorly defined. This study sought to define the contribution of CD4⁺ T_RM cells to pulmonary fibrosis, their origin, and regulatory mechanisms. We combined bioinformatic analysis of human fibrotic lung single-cell RNA-sequencing data with experiments in a bleomycin-induced C57BL/6 mouse model. Flow cytometry, targeted in vivo depletion, lymphocyte trafficking blockade, cell co-culture, and pharmacological inhibition were employed. CD4⁺ T_RM cells were observed at higher frequencies within fibrotic lung tissue. Their presence correlated with disease severity, and they were found to exhibit a pro-inflammatory and pro-fibrotic phenotype. Their specific depletion alleviated fibrosis. These cells primarily originated from recruited circulating lymphocytes, as blocking this recruitment reduced T_RM accumulation and attenuated disease. Furthermore, the Notch signaling pathway was activated in fibrotic lung CD4⁺ T_RM cells, and its inhibition suppressed their differentiation and impaired their pro-fibrotic function. We conclude that CD4⁺ T_RM cells are pathogenic drivers in pulmonary fibrosis, originating from circulating precursors and being regulated by Notch signaling, underscoring their relevance for therapeutic intervention. Full article