Journal of advanced research最新文献

Introduction: Rapeseed (Brassica napus L.) is a major oilseed crop with considerable potential for improving saline-alkali soils, yet the molecular mechanisms underlying its salt tolerance remain unclear.

Objectives: This study investigates the functional roles and regulatory mechanisms of glycine-rich protein 3 (BnaGRP3) in rapeseed under salt stress.

Methods: We employed molecular genetics, phenotypic and biochemical evaluation of transgenic rapeseed and Arabidopsis, transcriptome sequencing, protein interaction assays including immunoprecipitation-mass spectrometry (IP-MS), yeast two-hybrid (Y2H), luciferase complementation (LCA), and bimolecular fluorescence complementation (BiFC) assays, gene expression analysis by RT-qPCR, and hydrogen peroxide (H₂O₂) permeability assays conducted in yeast.

Results: BnaGRP3 was induced by salt stress and enhanced salt tolerance. Transcriptome analysis revealed that BnaGRP3 modulated expression of ion transporters under salt stress, especially NHX1 and SKOR. BnaGRP3 physically interacted with four plasma membrane intrinsic proteins (BnaPIPs). Overexpression of these BnaPIPs improved salt tolerance in Arabidopsis and increased H₂O₂ tolerance when expressed in yeast. In addition, these BnaPIPs formed both homomeric and heteromeric complexes, suggesting they may facilitate H₂O₂ permeability.

Conclusions: BnaGRP3 enhances salt tolerance by maintaining Na⁺/K⁺ homeostasis and, through its interactions with BnaPIPs, may participate in the regulation of H₂O₂ balance·H₂O₂ potentially serves as a bridge linking BnaGRP3-mediated ion homeostasis and redox regulation. The previously uncharacterized BnaGRP3-BnaPIP module broadens the mechanistic framework of GRP-mediated salt stress responses, thereby expanding our understanding of salt tolerance mechanisms in Brassica napus.

Introduction: Viruses are abundant biological entities within the gastrointestinal tract (GIT) of ruminants. Current understanding is extensive for bacterial and archaeal communities, but limited for viral communities.

Objectives: The study aimed to investigate viral diversity, virus-host interactions and ecological functions of viruses across GIT regions and ruminant species.

Methods: We collected 373 short-read and long-read metagenomes from 10 GIT regions of seven ruminant species, combining Illumina, PacBio HiFi, and Nanopore sequencing. Viral contigs were identified using sequence homology, viral hallmark gene and machine learning, and employed to uncover community assembly of spatial heterogeneity by analyzing virus-host linkage, lifestyle, and auxiliary metabolic genes (AMGs).

Results: We constructed a Ruminant Gastrointestinal Virome Catalog (RGVC) comprising 43,981 vOTUs, revealing that viral communities were remarkably diverse and mainly driven by the GIT regions rather than by the ruminant species. Virus-host linkage analysis identified 4603 putative prokaryotic hosts across 34 classes for 5954 host-linked viruses, along with robust correlation (R² = 0.91) observed between abundances of prokaryotic hosts and host-linked viruses across GIT regions. The lysogenic lifestyle was a dominant feature, with integrases being the predominant lysogenic-specific genes. We identified 864 high-confidence AMGs in lysogenic viruses that are annotated as key genes for polysaccharide degradation, glycolysis, and the Wood-Ljungdahl pathway, indicating a putative role for the viruses in supporting these host metabolic functions. The metabolic features of host-linked viruses were further verified by genomic context of selected AMGs of GH10, GPI and FHS with target function.

Conclusion: These findings suggest that the GIT viral communities exhibit spatial heterogeneity with distinct virus-host interactions, and offer new perspectives on maintenance of complex ecological and nutritional functions in ruminant GIT.

Background: Tryptophan (Trp) catabolism has been recognized as a key immunosuppressive axis in cancer. However, this largely centered on indoleamine-2,3-dioxygenase 1 (IDO1). The clinical failure of IDO1 inhibitors has exposed the limitations of this reductionist view.

Aim of review: To re-synthesize current knowledge into a further understanding of Trp metabolism, and propose biomarker-guided, multi-node intervention strategies that can resurrect Trp metabolism as a precision immuno-oncology target. Key Scientific Concepts of Review: This review comprehensively describes the pathways of Trp metabolism in the human body and the key enzymes that can serve as therapeutic targets, thus proposing the possibility of multi enzyme combined inhibition. Second, we synthesize how Trp metabolites can modulate the functionality of immune cells, mainly T cells, within the tumor microenvironment, thereby affecting tumor immune surveillance and the efficacy of immunotherapy. Then we discuss how tumor cells manipulate Trp metabolic pathways to enhance their survival and metastasis. We also propose a new framework for targeting Trp metabolism, such as combining enzymes inhibitors or Aryl hydrocarbon receptor (AhR) antagonists with immune checkpoint blockade. By shifting from "IDO1-focus" paradigms to comprehensive metabolic interventions, we may release more potential of Trp modulation in cancer immunotherapy.

Introduction: Autoimmune uveitis (AU) is an autoimmune disease of the eye that can lead to irreversible vision loss. Current therapies are limited by suboptimal efficacy and substantial side effects, highlighting the urgent need for the discovery of novel therapeutic targets. Nicotinamide phosphoribosyltransferase (NAMPT) is a key enzyme controlling the NAD⁺ salvage pathway and also exerts immunoregulatory and anti-inflammatory effects. However, its role in AU remains unclear.

Objective: To investigate NAMPT's effects on AU and underlying mechanisms.

Methods: Single-cell RNA sequencing (scRNA-seq) was performed on cervical draining lymph node (CDLN) cells from normal, experimental autoimmune uveitis (EAU), and NAMPT inhibitor-treated EAU mice. The influence of NAMPT inhibition on immune cell subsets, transcriptional programs, and intercellular communication networks was comprehensively analyzed. Additionally, scRNA-seq was performed on peripheral blood mononuclear cells (PBMCs) collected from Vogt-Koyanagi-Harada (VKH) disease patients and healthy controls (HC) to assess NAMPT expression and its modulation in human CD4⁺ T cells. In vivo and in vitro experiments, flow cytometry, and adoptive transfer experiments confirmed NAMPT's role in uveitis.

Results: NAMPT inhibition significantly ameliorated the clinical and histopathological manifestations of EAU. scRNA-seq revealed that NAMPT blockade reshaped immune cell composition and reversed disease-associated transcriptional programs, particularly within CD4⁺ T cells. It suppressed pro-inflammatory T helper (Th)-17 and Th1 responses while promoting regulatory T cell (Treg) populations. Mechanistically, NAMPT inhibition modulated the Th17/Treg balance by downregulation of Hif1α expression. In VKH patients, CD4⁺ T cells exhibited elevated NAMPT expression, which led to increased Th17 and Th1 cells and reduced Tregs. NAMPT knockdown reproduced the protective phenotype observed with FK866 treatment, suggesting a conserved NAMPT-Hif1α axis in human uveitis.

Conclusions: Inhibiting NAMPT can reverse the imbalance of effector T (Teff)/Treg cells by suppressing the expression of Hif1α in CD4⁺T cells, thereby effectively alleviating the symptoms of EAU. Therefore, NAMPT might be a potential target for AU.

Introduction: The widespread presence of antibiotic pollutants, such as tetracycline hydrochloride (TCH), causes significant environmental and public health concerns. Biochar-based photocatalysts derived from renewable biomass have attracted increasing attention due to their low cost, structural tunability, and environmental sustainability. However, their photocatalytic performance is often limited by poor charge separation and a lack of active sites.

Objectives: This study aims to construct a visible-light-responsive Cu/Fe co-doped biochar composite using Sphagnum palustre as a biomass precursor for the synergistic adsorption and photocatalytic removal of TCH from aqueous environments.

Methods: The Cu/Fe co-doped photocatalyst (CFO/S) was synthesized via a hydrothermal method by integrating Cu-Fe oxides with Sphagnum-derived biochar. The composite was comprehensively characterized, and its visible-light performance was evaluated. The photocatalytic mechanism was elucidated through radical trapping experiments and DFT+U simulations.

Results: The CFO/S-10 composite achieved a TCH removal efficiency of 94.56% within 60 min under visible-light irradiation. Adsorption was identified as the primary removal mechanism, while photocatalysis contributed to the degradation of adsorbed molecules. A layered FeO/CuFe₂O₄/S structure promoted charge separation and intermediate desorption. Multiple degradation products were detected, involving demethylation, hydroxylation, and ring-opening reactions.

Conclusion: The Cu/Fe co-doped biochar composite exhibited excellent removal performance through a synergistic adsorption-photocatalysis mechanism. Photogenerated electrons were the dominant reactive species, supported by •OH, •O₂^-, and h⁺. An S-scheme charge transfer mechanism was proposed to explain the enhanced redox capability. These findings demonstrate the potential of CFO/S as a promising candidate for visible-light-driven removal of antibiotic contaminants in water.

Background: Edible and medicinal mushroom polysaccharides (EMMPs) display wide-ranging bioactivities, yet progress has been limited by siloed workflows. Extraction, structural analysis, and biological evaluation are often conducted independently, obscuring how processing parameters shape polymer architecture and, consequently, biological function. This fragmentation contributes to inconsistent outcomes and poor reproducibility across studies.

Aim of the review: To bridge these gaps, this review (2020-2025) applies an extraction-driven structure-activity relationship (ESAR) framework that directly links processing conditions to structural features, mechanisms, and functional outcomes. It prioritizes well-characterized β-glucans where defined structural features allow direct mapping from extraction parameters to molecular architecture and biological effects. The objective is to shift the field from "finding an active extract" to engineering process-defined polymer architectures that deliver targeted mechanisms and reproducible, application-specific outcomes. Key scientific concepts of the review: Within ESAR, extraction variables such as solvent system, temperature, pH, and enzymatic or physical assistance influence branching patterns, molecular-weight distributions, and conformational features such as triple-helix stability. These structural attributes in turn influence bioactivity by governing receptor engagement and activation pathways. Comparative analyses across representative β-glucans reveal that differences in molecular weight ranges, branching patterns, and helix/coil conformations account for the distinct potencies observed across studies. It also clarifies common sources of variability related to strain differences, cultivation substrate, processing severity, and analytical methods. This review introduces ESAR as a unifying framework that converts fragmented polysaccharide studies into predictive design principles for real-world translation. It demonstrates how extraction-defined molecular engineering can drive reproducible development of functional foods, nutraceuticals, and adjuvant therapeutics. Such reproducibility is essential for translating laboratory findings into reliable industrial and clinical applications.

Introduction: Idiopathic pulmonary fibrosis (IPF) is a fatal interstitial lung disease with limited therapeutic options, thus necessitating novel strategies targeting upstream fibrogenic drivers; the exact impact of apolipoprotein E (apoE) on IPF and its therapeutic potential remain unexplored.

Objectives: This study aims to identify novel therapeutic targets for pulmonary fibrosis and elucidate the mechanism by which plasma apoE alleviates this condition.

Methods: We conducted an integrated meta-analysis of seven plasma cohorts and two-sample Mendelian randomization to assess apoE's association with IPF risk. CRISPR-engineered APOE-deficient canines and Apoe^‒/‒ mice were studied for pulmonary fibrosis. Mechanistic studies employed single-cell transcriptomics to identify fibroblast-enriched apoE receptors and SPIDER technology coupled with surface plasmon resonance (SPR) to characterize novel apoE interactors. Therapeutic potential was tested using the LXR agonist RGX-104 in murine models and human precision-cut lung slices.

Results: Plasma apoE was identified as a robust protective factor against IPF, with genetically elevated levels correlating with improved pulmonary function, and its deficiency in plasma showed potential diagnostic value for IPF. APOE-deficient canines developed spontaneous pulmonary fibrosis, and Apoe^‒/‒ mice exhibited exacerbated bleomycin-induced pulmonary fibrosis, reversible by tail vein injection of recombinant apoE protein. Fibroblast-specific enrichment of LRP1 and identification of PLAU as a high-affinity apoE interactor were observed. Mechanistically, apoE suppressed TGF-β/Smad-driven fibroblast activation via dual LRP1/PLAU co-engagement, attenuating α-SMA, collagen 1, and fibronectin. Pharmacological LXR activation (RGX-104) rescued apoE expression, reduced collagen deposition in vivo, and mitigated fibrosis in human precision-cut lung slices.

Conclusions: Plasma apoE is a causal guardian against pulmonary fibrogenesis, inhibiting TGF-β/Smad signaling through dual receptor (LRP1/PLAU) engagement. Cross-species validation and mechanistic elucidation position RGX-104, a small-molecule LXR agonist, as a potential therapeutic candidate for clinical translation in IPF.

Introduction: The eukaryotic translation initiation factor 4E (eIF4E) has emerged as a compelling target for cancer therapeutics due to its pivotal role in regulating cap-dependent translation of oncogenic mRNAs and its implication in various malignancies. However, the clinical potential of current eIF4E inhibitors is limited by suboptimal potency and binding affinity.

Objectives: Based on an analysis of the eIF4E/eIF4G binding pocket and structural features of existing inhibitors, 75 compounds were designed, synthesized, and screened. The binding affinity, molecular mechanism and antitumor activity of the most potent compound b14 were evaluated in vitro and in vivo.

Methods: Through structure-activity relationship analysis, 75 thiazole derivatives were synthesized and screened for binding affinity using fluorescence polarization (FP) and surface plasmon resonance (SPR). Hit compounds were evaluated for antitumor activity using the SRB assay. The most promising compound, b14, was further investigated for its antitumor activity and molecular mechanism via Western blotting (WB), quantitative real-time PCR (qRT-PCR), immunofluorescence, co-immunoprecipitation, and proteomics. The in vivo antitumor activity and safety of b14 were assessed using HeLa xenograft models and acute/subacute toxicity models, respectively.

Results: Compound b14 emerged as a lead molecule, exhibiting a 10-fold higher binding affinity to eIF4E than the reference inhibitor 4EGI-1. Mechanistic studies revealed that b14 disrupts eIF4F complex formation by inhibiting AKT-mTOR-4EBP1 and ERK-eIF4E phosphorylation, subsequently triggering mitochondrial dysfunction and apoptosis in tumor cells, with relatively low IC₅₀ values. Moreover, proteomics analysis further demonstrated that b14 suppresses oncogenic lipogenesis by downregulating key enzymes involved in lipid metabolism. Finally, oral administration of b14 significantly inhibits HeLa xenograft growth in vivo without measurable side effects.

Conclusions: Together, our results demonstrate that b14 is an excellent novel small-molecule inhibitor of eIF4E for future cancer therapy.