Search Results (35,605)

This study examined how 1-hydroxypyrene (1-OHP) affects bladder cancer (BCa) progression and tumor-associated immunosuppression. Public transcriptomic data and target-prediction databases were used to screen for targets related to 1-OHP and BCa. Machine learning was then used to narrow down the candidate genes. Molecular docking and single-cell RNA sequencing were also used to examine the selected targets and related pathways. The role of the main target was tested in vitro. CCK-8, colony formation, EdU, and Transwell assays were used to assess cell growth, migration, and invasion. Protein expression was examined by Western blotting. A tumor cell–CD8⁺ T-cell co-culture system was used to evaluate changes in CD8⁺ T-cell function. We identified 29 targets shared by 1-OHP and BCa. Machine learning further narrowed these targets to six genes, and macrophage migration inhibitory factor (MIF) was chosen for experimental validation. Treatment with 1-OHP increased BCa cell viability, proliferation, migration, and invasion. Silencing MIF partly reduced these effects. 1-OHP also increased MIF and CD74/CD44 expression and enhanced PI3K/AKT/mTOR signaling. These changes were reduced after MIF silencing. In the co-culture system, 1-OHP-treated BCa cells reduced CD8⁺ T-cell proliferation and function. Ki67, GZMB, and IFNG levels were decreased. MIF knockdown partly reversed these changes. Collectively, our findings show that 1-OHP promotes bladder cancer progression and tumor-associated immunosuppression, at least in part, through a MIF-dependent mechanism. By linking environmental exposure to activation of the CD74/CD44–PI3K/AKT/mTOR axis and impaired CD8⁺ T-cell function, our study provides new insight into the molecular basis of environmentally associated bladder carcinogenesis. Full article

Background: Differences in gene expression between inflammatory-like (iEOS-like) and resident-like (rEOS-like) eosinophil subtypes, and in eosinophil-derived serum mediators, may reflect eosinophil functional activity and their potential role in the pathogenesis of allergic asthma (AA). Methods: Twenty-three patients with non-severe AA and thirteen healthy subjects (HS) were examined. AA patients underwent a bronchial allergen challenge (BAC) with Dermatophagoides pteronyssinus and were re-evaluated 24 h later. Blood eosinophils were isolated by gradient centrifugation and magnetic separation, followed by subtyping based on CD62L expression. Gene expression was assessed by TaqMan-based quantitative PCR. Serum eosinophil cationic protein (ECP), eosinophil-derived neurotoxin (EDN), Galectin-10 (Gal10), and NADPH oxidase 2 (NOX2) were measured using ELISA. Results: Blood eosinophil subtypes from AA patients showed significantly higher expression of CLC, ECP, EPX, EDN, MBP, ALOX5, NOX2, and TGF-β1 compared to those from HS (p < 0.05), with no significant changes in LTA4H and LTC4S. iEOS-like cells in AA patients exhibited higher CLC and EPX expression compared to rEOS-like cells, (p < 0.05). Following BAC, CLC, MBP, and TGF-β1 expression increased in both eosinophil subtypes, while EPX and NOX2 increased only in iEOS-like cells (all p < 0.05). Serum ECP, EDN, and Gal10 concentrations were elevated in AA compared to HS and further increased after BAC (all p < 0.05); serum NOX2 remained unchanged. Conclusions: BAC induces a late-phase eosinophilic response in AA characterized by a partially eosinophil subtype-specific increase in gene expression and in circulating eosinophil-derived mediators. An increase in Gal10, observed at the CLC transcript level in eosinophils and in serum—but not in ECP or EDN—suggests the existence of mediator-specific mechanisms that regulate gene expression and extracellular release. Full article

In rapidly expanding African cities, the ecotoxicological relevance of PM₁₀ remains largely unresolved, especially where particle mass, chemical composition and emission sources are rarely assessed together. Here, 116 daily PM₁₀ samples collected in Luanda between 27 June and 5 November 2023 were tested using the Microtox^® Aliivibrio fischeri assay on aqueous extracts. Chemical speciation was combined with centred log-ratio (CLR) compositional clustering to identify recurring chemical regimes, while Positive Matrix Factorisation (PMF) was used to resolve the corresponding emission sources. Toxicity was moderate but highly variable: Toxic units after 15 min (TU₁₅) ranged from 0.84 to 8.54, with changes in toxicity units between 5 and 15 min (ΔTU) varying from −1.51 to +4.84. Only 3 of the 116 samples showed TU₁₅ < 1, and none reached TU₁₅ ≥ 10. The strongest responses occurred under a metal-enriched chemical regime (C2), characterised by elevated Zn, Pb and Cd concentrations, which showed the highest mean TU₁₅ (4.22) and ΔTU (1.52). This regime coincided with PMF evidence identifying non-ferrous metallurgy plus galvanised/metal scrap handling and burning as the factor most strongly associated with toxicity (ρTU₁₅ = 0.63; ρΔTU = 0.78), followed by a Ni-rich metallurgical factor with possible aviation influence (ρTU₁₅ = 0.51). Conversely, a marine/ionic chemical regime (C3) showed lower, stable toxicity, consistent with negative marine aerosol associations. These findings show that integrating chemical regimes with PMF-resolved sources identifies metal-related emissions as the main drivers of PM₁₀ ecotoxicity. Full article

Background: Pancreatic adenocarcinoma (PDAC) remains one of the most lethal malignancies, with limited survival gains despite advances in systemic therapy. However, rapid expansion of biomarker-directed therapies, immunotherapy, and novel treatment modalities has created an increasingly complex clinical trial landscape. We aimed to characterize contemporary PDAC clinical development and identify emerging therapeutic trends. Methods: We performed a narrative review with a scoping approach of active PDAC clinical trials registered on ClinicalTrials.gov. Eligible studies were initiated between January 1, 2021, and February 16, 2026, and included recruiting, active, or not-yet-recruiting phase I–III trials. Data extracted included trial phase, disease setting, therapeutic strategy, endpoints, enrollment, sponsorship, and late-phase development. Results: A total of 355 interventional trials were included. Most trials were phase I, phase I/II, or phase II trials, with fewer than 10% being evaluated in phase II/III or phase III development. Advanced or metastatic disease was the predominant setting. Chemotherapy remained the most frequently incorporated treatment modality, while molecularly targeted therapies were evaluated in 167 trials. KRAS/RAS-directed approaches represented the largest targeted subgroup, although only daraxonrasib and setidegrasib reached phase III evaluation. Immunotherapy was evaluated in 139 trials, although only a limited number progressed to late-phase development, reflecting the immune-resistant biology of pancreatic adenocarcinoma. Additional areas of active investigation included Claudin 18.2-targeted therapies, MTAP-associated approaches, homologous recombination deficiency-directed strategies, CD73 inhibition, radiotherapy, local interventions, surgery-focused optimization, imaging-guided approaches, and supportive-care interventions. Industry organizations were listed as the lead sponsor for approximately half of all studies, while non-commercial organizations supported most remaining trials. Conclusions: The contemporary PDAC clinical trial landscape is characterized by broad therapeutic diversification but limited late-phase maturity. Chemotherapy remains the dominant treatment backbone, whereas KRAS/RAS-directed therapies have emerged as the most advanced precision oncology strategy. Future progress will likely depend on successful integration of biomarker-selected therapies with established multidisciplinary treatment approaches, including optimized systemic therapy, local-control strategies, and supportive care. Full article

Background: Congenital pulmonary airway malformation (CPAM) is a rare developmental disorder characterized by cystic lung lesions, yet its extracellular matrix (ECM) composition remains poorly understood. This study employed decellularization and data-independent acquisition (DIA) proteomics to compare ECM profiles between cystic (CPAM) and histologically normal non-diseased (ND) regions from the lungs of four patients. Results: The decellularized scaffolds retained their native architecture with minimal residual DNA (<50 ng/mg). Proteomic analysis revealed 431 differentially expressed proteins (DEPs), with 171 upregulated and 260 downregulated in CPAM. Key findings revealed CPAM-specific enrichment of collagens (COL4A6, COL4A2, COL10A1, COL21A1 and PIIINP), glycoproteins (SPP1, FRAS1, FREM1, FREM2, LTBP1 and FBLN7), ECM regulators (TENM2, ROR2 and OMD), and ECM-affiliated proteins (ANXA7), alongside downregulation of glycoproteins (VASN and ABI3BP), proteoglycans (PODN and MXRA7), ECM regulators (SCARA5, PAPPA, SAA4, CPXM1, CTSC, THY1, SERPINA6/A1/D1, BSG, LYVE1, ITIH4 and CD44), and ECM-affiliated proteins (LGALSL). Pathway analysis highlighted the dysregulation of TGF-β, PI3K–AKT, and mTOR signaling in CPAM and aberrant ECM–cell interactions in pathogenesis. We also evaluated their functional properties and investigated the impact of ECM-based hydrogels on recellularization. Conclusions: These findings provide a comprehensive proteomic atlas of CPAM ECM alterations, offering insights into disease mechanisms and potential therapeutic targets. Full article

While carbon dots (CDs) are promising green nanomaterials for sustainable agriculture, how they regulate cotton growth via physiological and metabolic reprogramming remains poorly understood. This study evaluated foliar CD application (0–200 mg·L⁻¹) to uncover dose–response patterns and metabolic drivers. CDs exerted a concentration-dependent impact, with 100 mg·L⁻¹ yielding optimal results. This application markedly boosted shoot and root biomass (fresh weight: 103.43% and 44.28%; dry weight: 56.03% and 40.39%) and enhanced specific leaf area by 52.51% over the control. CDs improved photosynthetic efficiency (elevated Pn, Gs, and E values, alongside increased chlorophyll a and carotenoids) and strengthened antioxidant defenses (enhanced leaf POD/SOD and root POD/CAT activities). Despite mild MDA increases indicating oxidative stress, accumulated proline and soluble sugars in roots suggested adaptive osmotic adjustment. These findings suggest that CDs maintain physiological homeostasis by modulating antioxidant defense and osmotic adjustment. Untargeted metabolomics identified 2440 metabolites. Phenylpropanoid biosynthesis and flavonoid biosynthesis were the most prominently enriched pathways based on KEGG enrichment results. K-means and correlation analyses revealed key metabolites linked to cotton biomass. Overall, CDs facilitate cotton development through synergistic physiological and metabolic reprogramming, underscoring their potential as innovative agricultural growth regulators. Full article

Cutaneous T-cell lymphoma (CTCL), including mycosis fungoides (MF) and Sézary syndrome (SS), is defined by malignant CD4+ T-cell clones that evolve within and remodel the skin and systemic immune compartments. For disease biology and immune remodeling, we prioritize evidence from human primary samples; for therapeutic implications, we integrate clinical-trial and translational biomarker studies. We organize evidence by compartment (skin vs. blood) and, within skin, by stage (patch/plaque vs. tumor). Early patch/plaque MF often shows features consistent with constrained inflammation: skin-resident memory T-cell (TRM)-like malignant clones are in a Th1-leaning microenvironment with relatively preserved cytotoxic surveillance. Progression to tumor-stage MF (and/or large-cell transformation) is frequently associated with clonal dominance, Th2 cytokine polarization, upregulation of checkpoint pathways, and remodeling of myeloid/fibroblast populations. SS is typically associated with leukemic clonal dominance, systemic Th2 skewing, and broad impairment of cytotoxic immune control. These trajectories support an interpretive framework that aligns immune-directed therapies with compartment- and stage-associated biology; these include strategies that (i) deplete malignant clones (e.g., CCR4, CD30, KIR3DL2 targeting), (ii) re-engage exhausted effectors (PD-1/PD-L1 axis), (iii) reprogram cytokine balance (IFN-γ, IL-12, extracorporeal photopheresis), or (iv) suppress malignant signaling programs with secondary immune effects (JAK/STAT inhibition). We highlight potential candidate predictive biomarkers at varying levels of maturity, most of which require prospective validation, including antigen density, compartmental tumor burden, tumor cell fraction, effector-cell substrate, and interferon/cytokine signatures. Full article

This image report aims to illustrate the diagnostic value of integrating various diagnostic modalities to distinguish angiomatoid fibrous histiocytoma (AFH) from malignant soft tissue tumors. A 33-year-old man presented with a painful palpable mass in the medial left thigh. Magnetic resonance imaging revealed a 27 × 23 × 20 mm intramuscular lesion in the vastus medialis with a mixed T1 signal, predominantly high T2/STIR signal, internal hyperintense foci, a capsule-like rim, septations, heterogeneous enhancement, and marked peritumoral edema extending beyond the apparent tumor margins. Because these findings raised concern for a malignant soft tissue tumor, wide excision was performed after biopsy suggestive of low-grade sarcoma. Histologically, the tumor comprised spindle to epithelioid cells arranged in fascicular and storiform patterns surrounded by a lymphoid cuff. Immunohistochemistry showed positivity for CD68, CD99, and EMA, partial desmin and S-100 expression, and a Ki-67 index of 15%. Differential diagnosis included AFH and a malignant peripheral nerve sheath tumor. FISH demonstrated EWSR1 rearrangement, while PCR testing for EWSR1–CREB1 and EWSR1–ATF1 was negative. Although the specific fusion partner could not be identified, an EWSR1 rearrangement, together with the histologic and immunohistochemical findings, was considered supportive but not definitive for AFH. AFH, a rare intermediate tumor with nonspecific imaging features and variable pathology, makes diagnosis challenging. This case emphasizes the need to consider AFH in the diagnosis of intramuscular tumors with disproportionate peritumoral edema and highlights the role of molecular analysis in confirming the diagnosis. Full article

The progression of tuberculosis (TB) from a latent infection to an active disease involves intricate modifications in host immune, inflammatory, oxidative, and metabolic pathways. Ferroptosis represents a distinct form of regulated cell death that requires iron and is associated with excessive lipid peroxidation and has been associated with tissue damage in TB; however, its connection with host transcriptional changes during active TB is not fully understood. This study sought to identify and externally validate immune–inflammatory hub genes among differentially expressed genes (DEGs) in active TB that overlap with a ferroptosis-associated gene set derived from FerrDb, employing an integrated transcriptomic and systems-biology methodology. Differential expression analysis of GSE37250 revealed 1015 DEGs in active TB compared to latent TB, comprising 585 upregulated and 430 downregulated genes, and 93 DEGs in active TB compared to healthy controls, including 65 upregulated and 28 downregulated genes. Intersection analysis identified 94 DEGs common to the active TB versus latent TB comparison and the ferroptosis-associated gene set, and eight DEGs common to the active tuberculosis versus healthy-control comparison and the same gene set, with no genes shared across all three sets. Functional enrichment of the 94 intersection genes underscored immune response, defense response, stress response, Toll-like receptor signaling, NOD-like receptor signaling, IL-17 signaling, TNF signaling, glutathione metabolism, neutrophil degranulation, cytokine signaling, and antimicrobial metal sequestration. Protein–protein interaction analysis followed by cytoHubba prioritization identified 10 hub genes: IL1B, TLR4, CXCL10, MMP9, CYBB, MPO, CD36, LCN2, S100A8, and LTF. Subsequent to outcome-independent probe selection, external validation in GSE28623 demonstrated significant positive differential expression of LCN2, S100A8, and LTF, while GSE62525 showed significant positive differential expression of IL1B, TLR4, MMP9, MPO, LCN2, and LTF. LCN2 and LTF were significantly upregulated in both validation datasets, indicating the strongest cross-dataset reproducibility. These results identify an immune–inflammatory transcriptional network intersecting with ferroptosis-associated genes in active TB. Notably, the transcriptomic findings do not confirm ferroptotic cell death but suggest candidate genes and biological processes for future experimental exploration. Full article

Lipid nanoparticles (LNPs) are an effective platform for delivering therapeutic payloads, but their use for targeting specific cell populations in vivo is limited by a lack of specificity. To address this issue, we developed an innovative approach for creating targeted LNPs based on the post-insertion of protein–lipid conjugates. By expanding the genetic code, a single p-azido-L-phenylalanine residue was specifically incorporated into a DARPin against murine CD8, enabling copper-free click-chemistry with dibenzocyclooctyne (DBCO) to functionalize the particle surface with DARPin conjugates. DARPin-LNP characterization demonstrated preservation of particle size and acquisition of a characteristic negative charge after DARPin insertion, confirming successful post-insertion. In vitro experiments using primary murine splenic T-cells showed that targeted LNPs selectively transfected CD8⁺ T-lymphocytes. These data indicate that the developed platform enables selective in vitro delivery of nucleic acids to cytotoxic lymphocytes, highlighting its potential relevance for designing safer immunotherapeutic strategies. Full article

Background: Retroviral vectors remain widely used for CAR T-cell manufacturing, including emerging multi-component engineering strategies that incorporate additional functional modules to enhance therapeutic performance. However, genome-wide profiling of integration patterns in CAR T cells subjected to multiplex retroviral engineering has not previously been reported. Methods: We performed genome-wide profiling of retroviral integration sites in CAR T-cell products derived from two targeting platforms (IL15-GPC3 and HER2). Each platform was evaluated under conditions with or without co-transduction of a retroviral vector encoding ADA1 and CD26. Experiments were conducted using cells obtained from a single donor with three technical replicates. Vector copy number (VCN), chromosomal distribution, genomic feature annotation, and integration hotspot analyses were conducted. Results: VCN levels were comparable across all CAR T-cell products and were not significantly affected by ADA1/CD26 co-expression. Integration events were broadly distributed across the genome with enrichment in gene-dense chromosomal regions. Chromosome 19 showed a prominent integration preference, with a secondary enrichment observed on chromosome 17. The majority of integration sites were located within intronic regions, with no enrichment observed in promoter regions. Recurrent integration hotspots were identified; however, no evidence of dominant clonal expansion was observed across conditions. Comparative analyses between ADA1/CD26 and control groups showed minimal differences in integration patterns. Conclusions: To our knowledge, this study provides the first comprehensive, genome-wide characterization of retroviral integration site profiles in CAR T-cell products manufactured by multi-vector co-transduction with an ADA1/CD26 module. By employing harmonized comparison with external reference datasets, we offer a directly benchmarked integration site characterization of multi-component CAR T-cell manufacturing. These findings establish a reference framework for integration site monitoring in next-generation CAR T-cell engineering systems and inform risk evaluation for emerging combinatorial vector strategies. Full article

Astragalus membranaceus (AM) has been reported to support immune function, but its epigenetic mechanism on immune restoration of cyclophosphamide (CTX)-induced immunosuppression remains unclear. To evaluate the recovery of immune function, we assessed the injury of splenic pathology, cytokines levels in serum, proportional changes in CD4⁺ and CD8⁺ T cells, and the recall response of Hspx-specific T cell. In addition, to define the underlying epigenetic regulatory mechanism, we measured the expression levels of DNA methyltransferases (Dnmt1, Dnmt3a and Dnmt3b) and the methylation status of the Carnitine palmitoyltransferase 1A (Cpt1a) gene. The results showed that the crude extract of AM repaired the immunosuppression induced by CTX. This is reflected in the alleviation of splenic pathological injury, the elevated proportions of CD4⁺ and CD8⁺ T cells, and the increased levels of serum IL-2 and IFN-γ. The crude extract of AM also enhanced the recall response of Hspx-specific CD8⁺ T cells; that is, the frequencies of the resulting IL-2⁺ and IFN-γ⁺ cells were significantly higher than those in the CTX + Vaccine group. All these results indicate that the adaptive immune response induced by the crude extract of AM is significantly enhanced. On the other hand, AM crude extract reversed the CTX-induced downregulation of Dnmt3a mRNA and restored CpG methylation of Cpt1a gene to a level comparable to the vaccine group at molecular level. Consistent with the elevated CpG methylation, CTX-induced upregulation of Cpt1a mRNA was also reduced. These findings reveal that AM extract enhances the antigen-specific functional recall responses of CD8⁺ T cells in immunosuppressed mice were associated with Cpt1a gene methylation, thereby establishing a translational link between traditional herbal medicine and modern epigenetics. Full article

Background and Objectives: Anti-hepatitis B core antibody (anti-HBc) is generally regarded as a lifelong serological marker of previous hepatitis B virus (HBV) infection. However, anti-HBc seroclearance has been increasingly reported in patients receiving anti-CD20 therapy, while its incidence, longitudinal serological dynamics, and clinical significance remain insufficiently characterized. In this study, we aimed to evaluate the incidence, timing, and predictors of anti-HBc seroclearance and investigate longitudinal changes in HBV serology in HBsAg-negative/anti-HBc-positive patients undergoing anti-CD20 therapy. Materials and Methods: This retrospective single-center cohort study included 230 HBsAg-negative/anti-HBc-positive adults who received anti-CD20 therapy between 2011 and 2025. Anti-HBc seroclearance was evaluated using Kaplan–Meier analysis and Cox proportional hazards regression. Longitudinal changes in anti-HBc and anti-HBs titers were assessed using linear mixed-effects models. Results: Confirmed anti-HBc seroclearance occurred in 38 of 230 patients (16.5%). Younger age (adjusted hazard ratio (aHR) 0.96, 95% confidence interval (CI) 0.93–0.98; p = 0.001), fewer anti-CD20 treatment cycles (aHR 0.90, 95% CI 0.83–0.99; p = 0.025), and mycophenolate mofetil therapy (aHR 2.52, 95% CI 1.14–5.56; p = 0.022) were independently associated with anti-HBc seroclearance. Anti-HBc titers progressively declined during follow-up, with a significantly steeper decline among patients receiving mycophenolate mofetil. HBV reactivation occurred in five of 230 patients (2.2%), including two who had developed anti-HBc seroclearance. HBsAg seroreversion occurred in all five patients. Conclusions: Anti-HBc seroclearance is a relatively frequent serological finding in HBsAg-negative/anti-HBc-positive patients receiving anti-CD20 therapy and is accompanied by progressive declines in anti-HBc titers. Despite seroclearance, HBV reactivation may still occur, indicating that anti-HBc loss should not be considered evidence of elimination of HBV-related immunological risk. These findings improve our understanding of HBV serological dynamics during anti-CD20 therapy and support maintaining current antiviral prophylaxis and virological monitoring strategies regardless of anti-HBc seroclearance. Full article

Change detection in high-resolution optical remote sensing images is essential for dynamic surface monitoring and land resource management. However, existing methods still face challenges in representing multiscale change information, accurately locating change boundaries, and handling the imbalance between changed and unchanged samples. This study proposes SAM3TGNet, which transforms the general visual features of Segment Anything Model 3 (SAM3) into change-oriented representations through lightweight channel adaptation, multiscale cross-temporal interaction, and spatiotemporal feature fusion, while the Global Context Spatial Attention (GCSA) module and dynamic weighted loss enhance boundary representation and alleviate class imbalance, respectively. First, a SAM3-based bi-temporal remote sensing image feature encoder with channel adapters is developed, where intermediate multiscale feature layers are exploited to enhance semantic representation and generalization for diverse change patterns. Second, the GCSA module is introduced after spatiotemporal feature fusion to model global dependencies and enhance local details, improving boundary localization accuracy. Third, a dynamic weighted change loss function is designed to adaptively adjust the contribution of changed pixels according to their proportion in each batch, reducing background bias caused by sample imbalance and improving change localization and completeness. Experiments on the Wuhan University Change Detection (WHU-CD) and Sun Yat-sen University Change Detection (SYSU-CD) datasets demonstrate that the proposed method achieves F1-scores of 94.08 ± 0.12% and 82.91 ± 0.20%, respectively, outperforming existing approaches in multiscale change detection and boundary delineation. Full article

Telocytes are stromal cells characterized by small cell bodies and long cytoplasmic extensions termed telopodes. Although identified in the rat testicular interstitium, no standardized protocol for their isolation and in vitro enrichment has been established. This study aimed to develop an enriched primary culture model for rat testicular telocytes and characterize their morphology in vitro. Testicular cells from adult male Wistar rats were isolated by enzymatic digestion, subjected to short-term culture with differential adhesion, and enriched by fluorescence-activated cell sorting based on a CD34-positive/CD31-negative profile with empirical exclusion of events within the αSMA-associated region to reduce PTMC-like contamination. Post-sorting CD34 immunoreactivity, absence of detectable CD31 immunoreactivity, and characteristic morphology supported interpretation of the sorted fraction as telocyte-enriched. Light microscopy revealed recurring stellate, bipolar, compact branched, chain-like, and dense network-forming patterns. Some long, thin cytoplasmic processes showed a moniliform appearance with alternating thin segments and focal dilations. The findings demonstrate culture-associated morphological heterogeneity within the enriched population and provide a methodological framework for its isolation and light-microscopic characterization. Expanded post-sorting phenotyping, including PDGFRα and vimentin, together with ultrastructural validation, will be required in future studies. Full article