Search Results (31,236)

Background/Objectives: Local delivery of gene therapy products through the airways shows great promise for the treatment of a number of serious lung diseases, but its effectiveness is hampered by the mucus layer protecting the lung epithelium in the trachea and bronchi. Methods: To overcome this barrier, we engineered carbon dots (CDs) with mucus penetrating properties. Results: The CDs were synthesized by solvothermal treatment of citric acid and branched polyethyleneimine, and functionalized with maleamic acid groups to create cationic mucoinert nanoparticles with tunable charge. We characterized their interactions with a mucus model through turbidity and transport measurements, and assessed their impact on the viscoelastic properties of the biopolymer. We then demonstrated that the carriers are effective at delivering pDNA to a variety of cell models in vitro. In particular, mucus-producing Calu-3 cells cultured at the air–liquid interface (ALI) were used as a discriminating model to evaluate intracellular delivery of the genetic cargo through a thick layer of mucus at the cell surface. Conclusions: The functionalization of CDs with maleamic acid groups resulted in a 1000- to 10,000-fold increase in transfection efficiency in the mucus-producing model, offering new opportunities for lung gene therapy. Full article

Background: As the member of the B7 family, V-set and immunoglobulin domain-containing 4 (VSIG4) plays an essential role in regulating immune responses against bacterial infection, autoimmune disease, and chronic viral infection. However, the role of VSIG4 in acute viral infections remains largely unclear. Methods: Here, we constructed a gene-targeted VSIG4-deficient mouse model and then infected it with influenza to explore the detailed VSIG4-involved mechanism. Results: Our results demonstrated that the gene-deficient mice exhibited reduced survival rates, ranging from 25% to 50%, after being infected with different influenza virus strains. At the sites of infection, an increased number of CD8⁺ T cells, along with heightened expression of pro-inflammatory cytokines, e.g., Il-6 and TNFα, may have contributed to tissue damage. The recombinant VSIG4 protein slightly improved protection from the influenza challenge, suggesting regulatory functions of VSIG4 during infection. Using in vitro cell models, we show that the type C lectin receptor pathway member DC-SIGNR1 (CD209) is an essential factor during acute virus infection. The affinity and CO-IP tests indicated an interaction between CD209 and VSIG4, but not through protein modification. Conclusions: Therefore, VSIG4 functionally protected mice by regulating the type C lectin receptor pathway to inhibit excessive Th1 immune responses and inflammation. Our findings highlight the importance of considering immune homeostasis in the development of therapies for severe infections. Full article

Early B-cell development is primarily regulated at the transcriptional level and comprises the consecutive differentiation stages B-cell progenitor, pro-B-cell and pre-B-cell. These entities provide the cells of origin in B-cell precursor acute lymphoid leukemia (BCP-ALL) that show aberrations of developmental transcription factors (TFs), representing major oncogenic drivers. Analysis of physiological TFs in these developmental entities helps us to understand their normal and disturbed activities and regulatory connections. Here, we focused on NKL-subclass homeodomain TF NKX6-3, which is active in both normal B-cell progenitors and TCF3::PBX1 fusion gene-positive BCP-ALL cases. By performing siRNA-mediated knockdown and forced expression experiments in BCP-ALL model cell lines, we established a gene regulatory network for NKX6-3 together with TALE-class homeodomain TFs IRX1 and MEIS1, as well as ETS-TF SPIB. Importantly, NKX6-3 was activated by TCF3::PBX1, underlying their co-expression in BCP-ALL. Furthermore, comparative expression profiling analysis of public BCP-ALL patient data revealed TGFb-pathway in-hibitor CD109 as a downregulated target gene of NKX6-3. TGFb-signalling, in turn, enhanced NKX6-3 expression, indicating mutual activation. Finally, RNA-seq analysis of BCP-ALL cell line RCH-ACV after NKX6-3 knockdown revealed MPP7 as an upregulated target gene of both NKX6-3 and TCF3::PBX1, revealing a role for the HIPPO-pathway in B-cell progenitors and TCF3::PBX1-positive BCP-ALL. Collectively, our data introduce novel players and regulatory connections to normal and aberrant TF-networks in B-cell progenitors to serve as potential diagnostic markers or therapeutic targets. Full article

Background: Vitamin D has a significant role in immune system regulation due to its profound impact on various immune cells, including Natural Killer T-like (NKT-like) cells. While previous studies have explored the effects of vitamin D on the overall NKT-like cell population, detailed investigations into its impact on specific NKT-like subpopulations are lacking. This study aimed to analyze the correlation between vitamin D levels and NKT-like cell subpopulations (CD4+CD8+; CD4-CD8+; CD4+CD8-; CD4-CD8-) in peripheral blood collected from patients without diseases that can influence vitamin D and/or calcium levels. Methods: Peripheral blood mononuclear cells (PBMCs) were isolated from 86 patients. NKT-like cells were separated from PBMCs using a CD3+ CD56+ NKT cell isolation kit and a magnetic bead separator. Flow cytometry (FC) was applied in order to evaluate the distribution of NKT-like cell subpopulations. Results: A significant positive correlation between vitamin D levels and the CD4-CD8+ NKT-like cell population, particularly the CD4-CD8high subtype was found. Importantly, this correlation was independent of calcium levels, emphasizing the unique impact of vitamin D on CD4-CD8+ NKT-like cells. Conclusions: Our findings suggest that vitamin D concentrations may influence the distribution of NKT-like cell subpopulations in peripheral blood, although further evidence is necessary to confirm this observation. These novel results provide a foundation for elucidating the mechanism underlying the effect of vitamin D on the immune system and may contribute to future therapeutic strategies targeting CD4-CD8+ NKT-like cells in immune and oncological disorders. Full article

Background/Objectives: General anesthesia is occasionally required for cesarean delivery (CD). Propofol target-controlled infusion (TCI) enables dosing based on pharmacokinetic modeling. During the transition from induction to maintenance, infusion pauses. This simulation study assessed propofol from induction to delivery and the proportion of deliveries estimated during this pause. Methods: Surgical data from women undergoing CD were compiled, and the demographics were entered into a TCI pump using the Schnider model. Effect-site targets (6 and 8 mcg/mL) were simulated for induction, followed by 2.5 mcg/mL for maintenance. Outcomes were estimated propofol dose from induction to delivery and timing of delivery relative to infusion pause. Results: Among 50 women, the estimated mean propofol dose from induction to delivery was 19 ± 22 mg (0.2 ± 0.3 mg/kg) at 6 mcg/mL and 13 ± 17 mg (0.2 ± 0.2 mg/kg) at 8 mcg/mL. Delivery occurred during the infusion pause in 40% and 50% of cases, and it was more often in emergency than elective procedures. Emergency status, but not age or body mass index, predicted delivery during the pause. Conclusions: Standardized TCI with reduced effect-site targets for maintenance resulted in modest propofol administration between induction and delivery. These findings require confirmation in clinical studies, where dosing should be guided by depth-of-anesthesia monitoring. Full article

Exosomes are nanoscale extracellular vesicles that carry valuable biomolecular information. However, their characterization still depends on complex and costly techniques such as flow cytometry. In this study, a paper-based Vertical Flow Assay (VFA) specifically designed for the detection and profiling of exosomes derived from metastatic breast cancer cell lines is presented. The assay operates in an ELISA-like format, targeting exosomal surface proteins (CD9, CD63, CD81, and EGFR1) with specific antibodies and a secondary antibody conjugated to alkaline phosphatase. Upon reaction with the NBT/BCIP substrate, an insoluble indigo precipitate forms on the nitrocellulose membrane, generating a visual signal that can be further quantified by smartphone imaging. The VFA was optimized for membrane type, pore size, and blocking agents, reaching a detection limit of ~6 × 10⁷ exosomes µL⁻¹ in less than 20 min. Comparative studies with bead-based flow cytometry confirmed consistent biomarker expression profiles, demonstrating the reliability of the method. By enabling exosome biomarker profiling in a simplified and low-cost format, this approach provides a promising alternative to flow cytometry and other applications required for exosome characterization. Full article

Inflammatory bowel disease (IBD) is a chronic immune-mediated condition of the gastrointestinal tract, characterized by dysregulated inflammatory responses throughout the gastrointestinal tract. It includes two major phenotypes, Crohn’s disease (CD) and ulcerative colitis (UC), which present with varying gastrointestinal and systemic symptoms. The pathophysiology of IBD is multifactorial including genetic predisposition, mucosal and epithelial dysfunction, environmental injury, and both innate and adaptive immune response abnormalities. Several predisposing genetic factors have been associated with IBD explaining the strong hereditary risk for both CD and UC. For example, Caspase Recruitment Domain 9 (CARD9) variant rs10781499 increases risk for IBD, while other variants are specific to either CD or UC. CD is related to loss-of-function mutations in the nucleotide oligomerization domain containing the protein 2 (NOD2) gene and Autophagy-Related 16-like 1 (ATG16L1) gene. UC risk is increased particularly in Chinese populations by the A-1661G polymorphism of the Cytotoxic T-lymphocyte antigen 4 (CTLA-4) gene. This abnormal CTLA-4 interferes with B- and T-cell responses causing predisposition to autoimmune conditions. Previous studies suggested that IBD results from breakdown of the adaptive immune system, primarily of T-cells. However, new evidence suggests that a primary breakdown of the innate immune system in both CD and UC increases susceptibility to invasion by viruses and bacteria, with a compensatory overactivation of the adaptive immune system as a result. When this viral and microbial invasion continues, further damage is incurred, resulting in a downward cycle of further cytokine activation and epithelial damage. Released biomarkers also affect the permeability of the epithelial membrane, including lactoferrin, nitric oxide (NO), myeloperoxidase (MPO) and its activation of hypochlorous acid, matrix metalloproteinases (MMPs), especially MMP-9, omentin-1, and others. Increased macrophage and dendritic cell dysfunction, increased neutrophil activity, increased numbers of innate lymphoid cells, increased T-cells with decreased regulatory T-cells (Tregs), and changes in B-cell populations and immunoglobulin (Ig) functions are all associated with IBD. Finally, treatment of IBD has typically consisted of medical management (e.g., aminosalicylates and corticosteroids) and lifestyle modification, and surgical intervention in extreme cases. New classes of medications with more favorable side effect profiles include anti-integrin antibodies, vedolizumab, etrolizumab, and carotegrast methyl. Additionally, fecal microbiota transplant (FMT) is a newer area of research for treatment of IBD along with TNF-blockers, JAK inhibitors, and S1PR modulators. However, expense and long preparation time have limited the usefulness of FMT. Full article

This study presents the fabrication and performance optimization of porous fly ash-based geopolymer (FAGP)–polyethersulfone (PES) composite fibers with tunable FAGP loading for the multivariate adsorption of heavy-metal ions from aqueous solutions. Fibers containing 20 wt%, 40 wt%, and 60 wt% FAGP were prepared using phase inversion method and were characterized using X-ray computed tomography and mechanical testing. Adsorption experiments were conducted to assess the removal efficiencies of Pb²⁺, Cd²⁺, Cu²⁺, and Ni²⁺ at different pH values, temperatures, contact times, adsorbent dosage and initial metal-ion concentrations. The composite containing 60 wt% FAGP exhibited the high performance for all ions, and its performance was especially high for Pb²⁺. The isotherm and kinetic modeling revealed that the adsorption process followed Freundlich and Redlich–Peterson models, with mixed chemisorption–physisorption mechanisms depending on the metal-ion type. Compared with conventional adsorbents, the optimized composite fibers exhibited high adsorption capacity, enhanced handling suitability, and scalability in addition to their sustainability owing to the use of industrial by-products as precursors. These findings provide new insights into the structure–function relationships of FAGP composite fiber adsorbents and their potential for wastewater treatment applications. Full article

27-Hydroxycholesterol (27OHChol), a cholesterol metabolite, induces inflammatory responses in monocytic cells and promotes their differentiation into mature dendritic cells. Here, we examined whether inhibition of heat shock protein 90 (HSP90) modulates these responses. Treatment with ganetespib, a selective HSP90 inhibitor, significantly reduced chemokine CCL2 expression, lowering monocytic cell migration. It also suppressed matrix metalloproteinase-9 (MMP-9) expression and attenuated the lipopolysaccharide (LPS) response otherwise amplified by 27OHChol. Furthermore, ganetespib decreased mature dendritic cell markers (CD80, CD83, CD88) and restored endocytic activity, indicating a less activated state. These changes suggest that HSP90 regulates 27OHChol-induced pro-inflammatory activation via its client proteins. To explore this mechanism, we examined the phosphorylation status of signaling proteins. 27OHChol enhanced phosphorylation of Akt and its downstream targets, S6 and 4E-BP1 within the Akt/mTORC1 pathway. Ganetespib reduced total and phosphorylated Akt and 4E-BP1, and selectively inhibited S6 phosphorylation without altering total protein level. Collectively, these findings demonstrate that HSP90 inhibition by ganetespib mitigates 27OHChol-driven monocytic cell activation through suppression of the HSP90-Akt/mTORC1 axis. Targeting this pathway may provide a promising therapeutic strategy for metabolic inflammation associated with oxysterols. Full article

For patients with metastatic uveal melanoma (UM), tebentafusp is currently the only systemic therapy approved by the EMA and FDA, but its use is limited to HLA-A*02:01-positive individuals. Immune checkpoint blockade (ICB) represents another option, though only a small subgroup of patients benefits, and no reliable predictive biomarkers are available to date. The aim of this study was therefore to identify parameters associated with favorable ICB response. Tumor samples and clinical data from 30 patients were analyzed. Group A (n = 16) showed clinical benefit, while Group B (n = 14) experienced disease progression. NanoString^® analyses revealed 258 upregulated genes in Group A, including IDO1, CD28, and CCL8. The enriched pathways were predominantly linked to immune activation, leukocyte adhesion, and responses to external stimuli. Immunohistochemistry confirmed significantly higher CD28 expression on infiltrating immune cells in Group A, while a machine learning approach identified CCL8 as a predictive marker with ~78% accuracy. Overall survival differed significantly between the groups. These findings indicate that patients responding to ICB display tumors with enhanced immune activation. CD28 and CCL8 emerged as promising candidates and should be validated in prospective studies to determine their clinical utility. Full article

Tumor-infiltrating lymphocytes (TILs) are thought to play important roles in tumor shrinkage and survival prolongation in patients with breast cancer receiving neoadjuvant chemotherapy (NAC). TILs are mononuclear immune cells such as lymphocytes and plasma cells, including CD4+ and CD8+ T cells, natural killer cells, B cells, macrophages, regulatory T cells (Tregs), and myeloid/dendritic cells. The pre-NAC presence of more T cells and fewer Tregs in biopsy samples of primary breast tumors is known to contribute to tumor shrinkage and prolonged survival. This review was conducted to elucidate these roles in patients with breast cancer treated with NAC. Publications selected for inclusion in this review were identified by a PubMed search for articles published in English, performed using the terms “breast cancer”, “neoadjuvant chemotherapy”, “tumor-infiltrating lymphocyte”, “pathological complete response”, and “immune response”. The search was completed in July 2024. The functional roles of TILs in the achievement of these outcomes may vary by tumor subtype; increases and decreases in TIL levels before and after NAC have been shown to have conflicting effects. Biomarkers have been reported to predict local responses in the tumor microenvironment (e.g., immune-related gene signatures) and systemic immune responses (e.g., neutrophil-to-lymphocyte and platelet-to-lymphocyte ratios). Immune gene signatures and immune cell infiltration do not appear to be universally associated with tumor response or outcome in patients with breast cancer treated with NAC. The functional roles of TILs in breast tumor response and breast cancer survival may vary by tumor subtype, and conflicting results for the same subtypes may be due to differences in NAC regimens, immune responses, tumor heterogeneity, sample size, and the technical methods used to evaluate TILs in tumor samples. Full article

Accurate spatial information of cropland is crucial for precision agricultural management and ensuring national food security. High-resolution remote sensing imagery combined with deep learning algorithms provides a promising approach for extracting detailed cropland information. However, due to the diverse morphological characteristics of croplands across different agricultural landscapes, existing deep learning methods encounter challenges in precise boundary localization. The advancement of large-scale vision models has led to the emergence of the Segment Anything Model (SAM), which has demonstrated remarkable performance on natural images and attracted considerable attention in the field of remote sensing image segmentation. However, when applied to high-resolution cropland extraction, SAM faces limitations in semantic expressiveness and cross-domain adaptability. To address these issues, this study proposes a dual-branch framework integrating SAM and a semantically aware network (SAM-SANet) for high-resolution cropland extraction. Specifically, a semantically aware branch based on a semantic segmentation network is applied to identify cropland areas, complemented by a boundary-constrained SAM branch that directs the model’s attention to boundary information and enhances cropland extraction performance. Additionally, a boundary-aware feature fusion module and a prompt generation and selection module are incorporated into the SAM branch for precise cropland boundary localization. The former aggregates multi-scale edge information to enhance boundary representation, while the latter generates prompts with high relevance to the boundary. To evaluate the effectiveness of the proposed approach, we construct three cropland datasets named GID-CD, JY-CD and QX-CD. Experimental results on these datasets demonstrated that SAM-SANet achieved mIoU scores of 87.58%, 91.17% and 71.39%, along with mF1 scores of 93.54%, 95.35% and 82.21%, respectively. Comparative experiments with mainstream semantic segmentation models further confirmed the superior performance of SAM-SANet in high-resolution cropland extraction. Full article

Melanoma shows heterogeneity across body sites like skin, acral skin, and the uvea, driven by molecular characteristics and genetic variations. However, comparative studies exploring the heterogeneity of melanoma across different anatomical sites remain limited, hindering a comprehensive understanding of its underlying biology. We proposed a research framework through bioinformatics to analyze the tumor ecosystems of cutaneous, acral, and uveal melanoma, from molecular characteristics to genetic variations at single-cell resolution. We found that oxidative phosphorylation (OXPHOS) is a critical driver of tumor cell evolution, with abnormal ribosomal gene and tumor suppressor expression observed in uveal melanoma (UM). Additionally, we screened for potential drug targets and drugs against tumor cells. In the immune microenvironment, acral melanoma (AM) and UM exhibit stronger immunosuppressive characteristics compared to cutaneous melanoma (CM). OXPHOS contributes to T cell cytotoxicity dysregulation in CM and AM, while interferon-γ is crucial in UM. Tumor cells may also induce T cell dysfunction through biological signals such as MIF-CD74 and HLA-E-NKG2A. This study offers valuable insights into melanoma heterogeneity, providing a comprehensive research framework for understanding the distinct molecular and immune characteristics of CM, AM, and UM, and potentially guiding the development of therapeutic strategies tailored to each melanoma subtype. Full article

In-depth exploration of tumor immune suppression mechanisms may provide new therapeutic options for NF2-associated tumors. In this study, we found that sEVs secreted by NF2-associated schwannomas (NF2-EVs) facilitate the conversion of CD14⁺ monocytes into an MDSC-like phenotype, showcasing MDSC-like inhibitory functions. Moreover, these NF2-EVs are capable of enhancing tumor cell proliferation. Through proteomic analysis and subsequent validation of the NF2-EVs, we identified elevated levels of HSP90. When we knocked down HSP90 expression in tumor cells, the sEVs secreted showed diminished capacity to convert monocytes into MDSCs and a reduced ability to promote tumor cell proliferation. Conversely, sEVs secreted by tumor cells that overexpress HSP90 displayed the opposite effects. Further mechanistic studies revealed that HSP90 could influence the expression of AKT/p-AKT and ERK/p-ERK. Our results suggest that NF2 tumor cells could regulate the AKT/p-AKT and ERK/p-ERK pathways to promote tumor cell proliferation and the formation of an immunosuppressive microenvironment by secreting sEVs’ HSP90, offering valuable insights into the involvement of HSP90 in exosome-mediated communication within the context of NF2-related schwannomatosis (NF2-SWN). This information has the potential to inform the design of effective immunotherapeutic protocols and offer new treatment options for NF2-SWN patients. Full article

Under the global climate change, variations in climatic elements such as temperature, precipitation, and sunshine duration significantly impact the growth, development, and yield formation of winter wheat. A precise understanding of the impact of climate change on winter wheat growth and the scientific use of meteorological resources are crucial for ensuring food security, optimizing agricultural planting structures and agricultural sustainability. This study uses statistical methods and focuses on the Beijing–Tianjin–Hebei region, utilizing data from 25 meteorological stations from 1961 to 2010 and winter wheat yield data from 1978 to 2010. Twelve refined indicators encompassing temperature, precipitation, and sunshine duration were constructed. Path analysis was employed to determine their weights, establishing a comprehensive climate indicator model. Results indicate: Temperature indicators in the region show an upward trend, with accumulated temperature of the whole growth period increasing at a rate of 61.1 °C·d/10a. Precipitation indicators reveal precipitation of the whole growth period rising at 3.9 mm/10a and pre-winter precipitation increasing at 4.2 mm/10a. Sunshine duration exhibits a declining trend, decreasing at 72.7 h/10a during the whole growth period. Comprehensive climate indicators decrease from south to north, with the southwest region exhibiting the highest tendency rate (18.41), while the central and southern regions show greater variability. This study provides scientific basis for optimizing winter wheat planting patterns and rational utilization of climate resources in the Beijing–Tianjin–Hebei region. It recommends prioritizing cultivation in western southern Hebei and improving water conditions in the central and northern areas through irrigation technology to support sustainable crop production. Full article