Postbiotics, defined as preparations of inanimate microorganisms and/or their components, have attracted increasing attention because of their stability and functional benefits. Although lactic acid bacteria (LAB) modulate immune responses and oxidative stress, their direct effects on host mitochondrial homeostasis remain poorly understood. In this study, we investigated whether postbiotics derived from Ligilactobacillus salivarius enhance mitochondrial robustness in porcine intestinal macrophages (IPIMs). To establish a model of disrupted mitochondrial redox homeostasis, mitochondrial dysfunction was induced using the mitochondrial complex III inhibitor Antimycin A (AMA). High-dose AMA induced mitochondrial reactive oxygen species (mtROS) accumulation and suppressed the expression of antioxidant genes, including SOD2. Pre-stimulation with heat-killed L. salivarius suppressed AMA-induced mtROS production in selected representative strains, with the most effective strains enhancing SOD2 expression. These mtROS-suppressive strains selectively activated Toll like receptor (TLR)1/2 and TLR2/6 signaling, whereas non-protective strains failed to induce TLR responsiveness. Pharmacological inhibition of TLR2 abolished mtROS suppression and SOD2 induction, confirming TLR2 dependency. Furthermore, Seahorse extracellular flux analysis revealed that postbiotics enhanced mitochondrial respiration, including maximal respiration and spare respiratory capacity, via TLR2 signaling. Collectively, our findings demonstrate that postbiotics function as immunometabolic modulators that reinforce mitochondrial redox homeostasis and respiratory capacity via TLR2 signaling, highlighting a novel mechanism by which microbial components contribute to intestinal immune cell homeostasis.
This multi-source bibliometric and translational mapping study provides a panoramic synthesis of how research on microglia-mediated spinal pain signaling has evolved from foundational mechanistic studies to clinically oriented innovations. The aim is to identify developmental trajectories, mechanistic hotspots, and translational opportunities, thereby offering strategic insight into guiding the future direction of neuropathic pain research. We analyzed 1313 original research papers from the Web of Science Core Collection (WoSCC; 2005-2024) using CiteSpace and VOSviewer to construct collaboration networks, journal co-citation graphs, and keyword-driven mechanism clustering. To add a translational medicine dimension, we conducted a targeted PubMed search ("microglia AND spinal cord AND (translational OR therapeutic OR drug targets)"), retrieving 692 additional records, enabling cross-database overlay to link mechanistic themes with specific therapeutic targets. The scientometric model indicates that spinal pain research has shifted from primarily descriptive work to more detailed regulatory models. Key themes include glial cell activation, oxidative stress, mitochondrial dysfunction, and changes in microglia state. Research on heat shock protein pathways and sex-related microglial responses is also increasing. Some core terms have remained frequent over the years, such as "neuroinflammation" and "activated protein kinases". In contrast, the explosive emergence of brain-derived neurotrophic factor (BDNF) and spinal cord stimulation (2020-2021; burst intensity = 2.56) indicates a growing interest in synaptic and circuit control and neuromodulation-based approaches. In the PubMed subset, 33.6% of studies directly focused on treatment development, with gene therapy, intrathecal administration, and microenvironment remediation also appearing more frequently. When we combine data from WoSCC and PubMed over the past 20 years, we can see a significant shift in the explanation of spinal pain in this field. Early research often described the problem as "glial cell activation-cytokine release." Recent research, however, focuses on specific pathways, particularly microglial state regulation, oxidative stress-autophagy connections, and kinase signaling. This shift in treatment approaches is also reflected in translational studies. Many studies no longer rely primarily on systemic drugs but instead focus on targeted strategies such as intrathecal administration, gene or cell therapy, extracellular vesicles, and neuromodulation. These trends make polarization-related molecular nodes ideal candidate targets for precision analgesia. However, bibliometric results are dependent on database coverage, keyword processing, and clustering settings. Some "hotspots" may reflect changes in terminology or citation habits rather than true mechanistic importance. The rise of neuromodulation keywords may also reflect broader clinical applications; microglial mechanisms are plausible, but contributions from other circuit-level mechanisms may also play a role. These results indicate that the field is moving beyond a purely inflammatory perspective toward systemic intervention models. Currently, there is a greater focus on microglial homeostasis and M2-like anti-inflammatory/immune repair processes, as well as sex and metabolic factors that may influence responses. This research direction supports immune repair and more personalized analgesia. Simultaneously, stronger mechanistic arguments require cell state-specific measurements rather than broad phenotypic labels.
Although fear conditioning has elucidated cue-evoked acute fear responses, the mechanisms by which stress experiences induce generalized internal states linked to anxiety or phobia are poorly understood. Here, we report that robust stress induces a persistent behavioral change characterized by avoidance of a confined space, claustrophobia-like behavior (CLB) in Drosophila. Unlike aversive memory formation, the development of CLB does not require dopamine receptors. Our neuronal screening determined that neuropeptide signaling via Allatostatin-A inactivates the downstream neurons via its receptor AstA-R1, causally inducing CLB. Moreover, gene expression profiling of individual fly heads revealed that innate immune response activation in the blood-brain barrier is involved in CLB. Our data demonstrate that stress-induced persistent behavioral change would not be related to a canonical mechanism of aversive memory formation, rather involves neuropeptidergic signaling and the blood-brain barrier, providing the mechanism determining internal states which persistently change into a phobia-like mode.
The major histocompatibility complex class II (MHC-II) pathway is central to adaptive immunity and immune tolerance, and its age-related dysregulation is increasingly linked to chronic neuroinflammation. The HLA-DRB1*15:01 allele, the strongest genetic risk factor for multiple sclerosis, has been implicated in shaping pathogenic CD4+ T-cell responses and broader neuroimmune vulnerability, yet how this allele modulates age- and sex-dependent neuroimmune processes within the central nervous system (CNS) remains poorly defined. We investigated the impact of HLA-DRB1*15:01 expression using a humanized mouse model (HLA mice) and wild-type (WT) controls. Male and female mice were analyzed at 6, 9, and 15 months of age, with endocrine stratification in females. Behavioral testing, flow cytometry, immunofluorescence, and multiplex cytokine analyses were used to assess cognitive performance, glial immune-associated changes and oxidative stress, astrocyte-microglia IL-3/IL-3R signaling, endothelial activation, selective immune cell accumulation at CNS borders, tissue organization, and hippocampal cytokine profiles. HLA mice developed age- and sex-dependent cognitive impairment, most pronounced in aged females. HLA-DRB1*15:01 expression promoted progressive microglial immune-associated changes, characterized by increased CD14 and CD68 expression, elevated mitochondrial oxidative stress, altered astrocyte phenotypes, and enhanced IL-3/IL-3R signaling. Hippocampal axonal and myelin organization was disrupted in aged HLA mice and was spatially associated with increased microglial presence. HLA mice also exhibited selective immune remodeling, including increased accumulation of CD4+ T cells and NK1.1+CD3+ natural killer T (NKT) cells, particularly in females, accompanied by endothelial activation marked by elevated ICAM-1 and E-selectin expression. Hippocampal cytokine profiling revealed selective sex-biased alterations, without broad induction of classical inflammatory cytokines. Together, these findings demonstrate that HLA-DRB1*15:01 drives a coordinated, age- and sex-dependent neuroinflammatory program linking behavioral dysfunction, glial immune-associated changes and oxidative stress, selective immune cell recruitment, endothelial activation, tissue remodeling, and targeted cytokine imbalance. This integrated phenotype provides mechanistic insight into how this major MS risk allele confers vulnerability to chronic neuroinflammation during aging, with heightened impact in females, independent of reproductive cycling stage.
DNA N6-methyladenine (6 mA) has recently been recognized as a novel epigenetic modification, with ALKBH4 identified as a specific demethylase. However, the role of ALKBH4 and DNA 6 mA in the pathogenesis of ossification of the posterior longitudinal ligament (OPLL) remains unclear. Tissue samples from OPLL patients and normal posterior longitudinal ligaments were collected, and ligament fibroblastic cells (LFCs) were isolated from OPLL tissues using primary culture. The expression and functional relevance of ALKBH4 in OPLL were investigated through reverse transcription quantitative polymerase chain reaction and western blot analyses. Osteogenic potential was further assessed using alizarin red S staining and alkaline phosphatase activity assays. Methylation status was evaluated by enzyme-linked immunosorbent assay and chromatin immunoprecipitation. Furthermore, the regulatory role of ALKBH4 in LFC ossification was investigated through the bone morphogenetic protein 2 (BMP2) and Wnt/β-catenin pathways. ALKBH4 expression was significantly elevated in both OPLL tissues and LFCs, whereas global 6 mA levels and BMP2-associated 6 mA were reduced. Overexpression of ALKBH4 promoted the ossification of LFCs, while its knockdown suppressed osteogenesis. ALKBH4-mediated DNA demethylation at the 6 mA site facilitated Yin Yang 1 (YY1) binding to the BMP2 promoter, enhancing BMP2 transcription and driving ossification. Silencing of BMP2, Wnt/β-catenin, or YY1 attenuated the pro-ossification effects of ALKBH4. Furthermore, the mutation that affects the action of ALKBH4 at the 6 mA site fails to induce the osteogenic effect of LFC. These findings demonstrate that ALKBH4 regulates OPLL progression by modulating BMP2 and Wnt/β-catenin signaling and by promoting BMP2 transcription through site-specific demethylation in human OPLL tissues and primary LFCs. ALKBH4 may therefore represent a promising therapeutic target for the management of OPLL.
In transfected cells, adhesion G protein-coupled receptors (GPCRs) are activated by tethered agonists that are embedded in their canonical autoproteolytic GAIN domain. It is unknown, however, whether a tethered agonist-dependent activation mechanism generally mediates the physiological functions of adhesion GPCRs. Here, we show that G protein signaling by BAI3 (Adgrb3), a brain-specific adhesion GPCR, is essential for its functions in controlling axon and dendrite growth and promoting synapse formation. Moreover, our signal transduction assays confirm that constitutive exposure of BAI3's tethered agonist massively stimulates (~5-fold) its GPCR activity. However, the constitutive exposure of BAI3's tethered agonist, produced by deletion of its extracellular domains, blocked instead of activating BAI3's functions in regulating axonal and dendritic growth and promoting synapse formation. Moreover, inactivating mutations of BAI3's tethered agonist or deletion of BAI3's constituent GAIN domain did not detectably impair BAI3's physiological functions. Thus, the GPCR activity of BAI3 is functionally required, whereas tethered agonist-mediated stimulation of its GPCR activity is not.
Autism spectrum disorder (ASD) encompasses a group of neurodevelopmental disorders influenced by genetic and environmental factors, although the molecular mechanisms underlying their interactions remain unclear. We previously reported dysregulated cytokine signalling in chromosome 15q-duplication syndrome (Dup(15q)), a common syndromic form of ASD. Dup(15q) induced pluripotent stem cell (iPSC)-derived neurons exhibit an amplified Signal Transducer and Activator of Transcription-3 (STAT3) response to Interleukin-6 (IL-6), a cytokine often upregulated in ASD. To identify candidate genes within the 15q region that may modify cytokine signalling, we investigated Cytoplasmic FMRP-Interacting Protein 1 (CYFIP1), as CYFIP1 dysregulation has been linked to altered expression of genes involved in immunoregulatory pathways. CYFIP1 was overexpressed using a plasmid vector in human HEK-293 and SH-SY5Y neuroblastoma cells. Following stimulation with IL-6 or Interferon (IFN)-γ, a variety of biochemical assays (qRT-PCR, Western blotting and dual-luciferase reporter assays) and neurite tracing experiments were performed to assess the effects of increased CYFIP1 expression on IL-6/STAT3 and IFN-γ/STAT1 signaling responses. CYFIP1-overexpression HEK-293 cells display reduced STAT3 and STAT1 expression, but enhanced IL-6-induced/IFN-γ-induced STAT3/STAT1 transcriptional activity. Furthermore, CYFIP1-overexpression in SH-SY5Y cells was associated with reduced basal neurite outgrowth and altered IL-6-associated neurite outgrowth. These findings suggest that CYFIP1-overexpression modifies cytokine-responsive transcriptional pathways in vitro and provides novel insight into how CYFIP1 dysregulation may contribute to dysregulated cytokine signaling in ASD, advancing our understanding of the molecular mechanisms underlying neuroinflammatory processes in this disorder.
Domestication has profoundly transformed human production and lifestyles. The Qingtian rice-fish co-culture system is the first globally important agricultural heritage system (GIAHS). PF-carp are a key species in the Qingtian rice-fish system and have been domesticated in rice paddies for more than a millennium, yet the mechanisms of their tolerance to high temperature conditions remain unresolved. In this study, 28℃ as the control group (C0), and two heat stress groups were established at 38℃ for 0 h (H0) and 24 h (H24). Brain tissues were sampled for physiological index measurements and transcriptomic analysis. Physiological analyses showed that the activities of SOD, CAT, and GSH-Px increased initially and then declined, whereas MDA levels exhibited a continuous increase. Transcriptome profiling identified 9,825 differentially expressed genes (DEGs). KEGG enrichment analysis of DEGs indicated that immune responses and metabolic regulation were consistently involved throughout the thermal adaptation process. During acute warming phase, pathways such as protein processing in the endoplasmic reticulum, FoxO signaling pathway and glycerophospholipid metabolism were enriched. Under prolonged high temperature exposure, cytokine-cytokine receptor interaction, PPAR signaling pathway and TGF-β signaling pathway were prominently enriched. Within these pathways, genes including grp94, hsp70, bip, nef, il-1r, tlr8, cctgα6, ccr4, cxcr3, and il-10 were significantly up-regulated (p < 0.05). These results indicate that PF-carp exhibit coordinated brain physiological and transcriptomic responses to high temperature exposure, involving protein quality control, immune signaling, and metabolic regulation. Collectively, our findings provide new insights into the mechanisms by which PF-carp adapt to thermal exposure and provide theoretical support for the breeding of heat tolerant fish.
Hepatocellular carcinoma (HCC) is a major health issue, but treatment options are limited. This study investigated the role of CCR4-NOT transcription complex subunit 9 (CNOT9) in the pathogenesis of HCC and its potential as a therapeutic target. Bioinformatics analysis was performed on RNA-seq data from the TCGA and GTEx databases to assess CNOT9 expression and prognostic significance. CNOT9 expression was validated in clinical samples and cell lines using qRT-PCR and Western blot. CNOT9 was knocked down in HCC cell lines, and its effects on proliferation, apoptosis, and cell cycle were investigated using functional assays (CCK-8, EdU, colony formation, and flow cytometry). The underlying molecular mechanisms were explored via RNA-seq and Western blot analysis of the AKT pathway and cell cycle regulators. Xenograft mouse models were used to confirm the oncogenic role of CNOT9 in vivo. CNOT9 mRNA and protein expression were upregulated in HCC patients and associated with poor prognosis. CNOT9 induces abnormal proliferation of HCC cells and G2/M phase cell cycle progression. Knocking down CNOT9 reduces cell proliferation, increases apoptosis, and causes cell arrest at the G2 phase. CNOT9 knockdown activates PTEN to inhibit the AKT pathway and suppresses the expression of cell cycle-related proteins p53, p21, CCNE1 and CDK2. CNOT9-deficient tumors exhibited reduced growth in mice, supporting its pro-oncogenic role. This study first elucidates the molecular mechanism by which CNOT9 drives HCC progression through post-transcriptional regulation of the PTEN/AKT/p53 axis, providing a theoretical basis for precision treatment strategies targeting CNOT9 or the PTEN/AKT/p53 pathway.
MDM2 is transcriptionally activated by the ST-MYCL-Tip60 complex in virus-positive Merkel cell carcinoma (MCC). MDM2 suppresses p53 and is a rational therapeutic target. MDM2 inhibitors face an intrinsic limitation: p53 activation induces MDM2 transcription, creating a feedback loop that blunts inhibitor efficacy. We demonstrate that MDM2 degraders KTX-049 and KT-253 overcome this limitation by collapsing the p53/MDM2 negative feedback loop. KTX-049 was >100-fold more potent than the MDM2 inhibitor DS-3032 across WT p53 MCC cell lines, and this superior potency was quantitatively supported by mechanistic mathematical modeling. In vivo, KT-253 produced deep and durable tumor regressions, including complete responses in patient-derived xenograft models. Acquired resistance was strongly associated with acquisition of TP53 mutations, confirming on-target pathway pressure. These findings establish feedback architecture as a critical determinant of therapeutic response and position MDM2 degradation as a qualitatively distinct strategy that produces more durable pathway engagement than MDM2 inhibition, providing a preclinical rationale for prioritizing MDM2 degraders in WT TP53 MCC.
This study aimed to explore the dental applications of AG73, a laminin-derived adhesive peptide, by examining its effects on human dental pulp cells (hDPCs), particularly in cell adhesion and mineralization. It also sought to identify the key functional residue of AG73 and the signaling pathways involved in its pro-mineralization activity. We compared AG73's pro-mineralization activity with peptides derived from LAMA5 using alizarin red staining. Immunofluorescence staining assessed its cell adhesion properties. Alanine-scanning mutagenesis was performed to identify key residues for AG73's adhesion activity. Finally, we used p38 and JNK MAPK inhibitors to determine which signaling pathways were involved in AG73-induced mineralization. AG73 exhibited superior pro-mineralization activity compared to other peptides derived from LAMA5 in hDPCs. Immunofluorescence staining confirmed AG73's strong cell adhesive ability within 20 minutes. Alanine-scanning identified isoleucine at the tenth position as crucial for adhesion. AG73 was the most effective peptide in inducing mineralization, with no mutated versions outperforming it. p38 signaling was found to play a positive role in AG73-induced mineralization, while JNK inhibition enhanced mineralization, suggesting JNK inhibitors as potential inducers. AG73 has strong pro-mineralization activity and significant cell adhesive properties in hDPCs. The isoleucine at the tenth amino acid position was identified as a critical residue for AG73's adhesive ability. Additionally, p38 was found to be involved in AG73-regulated mineralization, while JNK inhibition promoted the process. This is the first study to explore AG73's effects in hDPCs and provides a foundation for its potential use as a dental-pulp regenerative material.
To investigate the anti-fibrotic effects of Isofraxidin (IF) on liver fibrosis and its underlying mechanisms. C57BL/6J mice (bred by The Jackson Laboratory) were induced with liver fibrosis using CCl4 and treated with different doses of IF (20 mg/kg and 40 mg/kg) via gavage. Colchicine served as the positive control. Liver fibrosis markers (α-smooth muscle actin,α-SMA; Fibronectin, FN; Collagen type I, Col-I) were assessed by histological staining, immunohistochemistry, and western blotting. Network pharmacology and molecular docking were adopted to conjecture underlying targets and signaling pathways of IF, followed by experimental validation. IF at various doses improved liver function and exhibited dose-dependent anti-fibrotic effects. Network pharmacology identified Protein kinase Kinase B (PKB, Akt), Proto-oncogene tyrosine-protein kinase (Src), Epidermal growth factor receptor (EGFR), Heat Shock Protein 90 Alpha Family Class A Member 1 (HSP90AA1), Glycogen Synthase Kinase 3 beta (GSK3β), and Monoamine Oxidase B (MAOB) as the core targets. Molecular docking showed good binding affinity between IF and these targets. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses indicated that the Nuclear Factor kappa-light-chain-enhancer of activated B cells (NF-κB) signaling pathway is a major pathway involved in IF's anti-fibrotic effects. Experimental validation confirmed that IF inhibited the expression of inflammatory factors (Tumor Necrosis Factor-alpha, TNF-α; Interleukin-1 beta, IL-1β; Interleukin-10, IL-10) and the phosphorylation of NF-κB and Inhibitor of Nuclear Factor Kappa B Alpha (IκBα). IF effectively alleviated CCl4-induced liver fibrosis in mice by modulating key targets (AkT, Src, EGFR, HSP90AA1, GSK3B, and MAOB) and the NF-κB signaling pathway.
RNA helicases are a large family of enzymes crucial for virtually all aspects of RNA metabolism, forming the backbone of gene expression regulation. Among them, DEAH-box helicase 33 (DHX33) has emerged as a pivotal player in fundamental cellular processes, including ribosomal biogenesis, transcription, and translation initiation. A compelling body of evidence now positions DHX33 as a significant oncoprotein, with its overexpression documented in a wide spectrum of human cancers such as lung carcinoma, hepatocellular carcinoma, glioblastoma, and acute myeloid leukemia. Its oncogenic drive is mediated through the transcriptional regulation of genes governing the cell cycle and apoptosis, its interplay with major signaling pathways like Wnt/β-catenin and PI3K/Akt/mTOR, and its role in metabolic reprogramming, notably the Warburg effect. Furthermore, DHX33 acts as a key downstream effector of potent oncogenes like c-Myc. Genetic or pharmacological inhibition of DHX33 consistently impedes tumor growth, underscoring its non-redundant role in oncogenesis. This review systematically synthesizes the current understanding of the mechanisms by which DHX33 promotes tumorigenesis. It delves into its regulation of core cellular processes, its integration into oncogenic signaling networks, and its recently discovered functions in epigenetic and metabolic reprogramming. By consolidating this knowledge, we aim to highlight the multifaceted nature of DHX33 in cancer biology and firmly establish its potential as a viable and promising therapeutic target for future anticancer strategies.
Tea plant (Camellia sinensis) is an economically important crop whose yield and quality are threatened by anthracnose caused by Colletotrichum camelliae. While C. camelliae infection elevates indole-3-acetic acid (IAA) levels in tea plants, how this hormone increases influences disease progression remains unclear. Here, we show that C. camelliae actively rewires auxin homeostasis in tea plants to compromise immunity. Using functional genomics and molecular analyses, we identified an auxin transporter, CsWAT1-19, which is associated with intracellular auxin compartmentalization. Its pathogen-induced expression is essential for cytosolic free IAA accumulation and subsequent disease development. We further delineated a regulatory cascade in which pathogen infection downregulates the transcriptional repressor CsNF-YB4, thereby relieving its repression of CsWAT1-19. The resulting increase in cytosolic free IAA activates the auxin-responsive factor CsARF9, which directly suppresses key lignin biosynthetic genes and disrupts cell wall-based defense. Together, our findings define a CsNF-YB4→CsWAT1-19→CsARF9 regulatory module through which a fungal pathogen hijacks host auxin transport and signaling to attenuate lignin-mediated physical barriers, providing new insight into the intersection of plant hormone signaling and immune responses in tea plants.
Chronic graft-versus-host disease (cGVHD) is a major complication of allogeneic hematopoietic stem cell transplantation, with cutaneous fibrosis representing a debilitating and treatment-resistant manifestation. We identified a hypoxia-driven inflammatory and fibrotic program in cGVHD skin mediated by a hypoxia-inducible factor 1α-phosphatidylinositol 3-kinase-interleukin-13 (HIF-1α-PI3Kδ-IL-13) signaling axis. Spatial transcriptomics, single-cell RNA sequencing, and multiplex immunofluorescence revealed stabilization of HIF-1α in epidermal cells in hypoxic regions. HIF-1α stabilization was accompanied by IL-13 production from PI3Kδ-activated T cells and macrophage-epidermal cross-talk, which together promoted fibrosis and the formation of tertiary lymphoid structure (TLS)-like aggregates. In human epidermal cell lines and induced pluripotent stem cell-derived skin organoids, IL-13 directly induced HIF-1α expression in keratinocytes under normoxic conditions. HIF-1α expression was further amplified in hypoxic conditions, driving profibrotic signaling. Pharmacologic inhibition of HIF-1α, PI3Kδ, or IL-13 reduced tissue hypoxia, disrupted TLS-like aggregates, and ameliorated disease in murine cGVHD models. These findings identify a targetable circuit linking hypoxia, type 2 inflammation, and immune-stromal dysfunction in cGVHD and provide a translational framework for treating fibrotic cGVHD and related immune-mediated fibrosing diseases.
Probiotics hold promise for enhancing coral resilience under climate-driven thermal stress, yet their mechanisms remain poorly understood. Although the bacterial genus Endozoicomonas has been proposed to benefit corals, in vivo evidence of beneficial effects on the host remains limited. Here, we establish Endozoicomonas acroporae Acr-14T as a coral probiotic and characterize its effects on the reef-building coral Stylophora pistillata. We show that E. acroporae Acr-14T enhances host thermal tolerance, colonizes coral tissues, and forms coral-associated microbial aggregates (CAMAs). Microbial profiling indicates that probiotic treatment is associated with reduced relative abundances of opportunistic microbes and enrichment of putatively beneficial taxa. To support transcriptomic analyses, we assembled a chromosome-level genome of S. pistillata clade 1 (Pacific lineage) and found that E. acroporae Acr-14T treatment mitigates heat-induced protein-folding stress and apoptotic signaling. Single-cell transcriptomics further revealed altered expression of genes involved in S-adenosylmethionine (SAMe) metabolism and pro-survival signaling in gastrodermal cells of probiotic-treated corals. Together, our results provide a cell-type-resolved view of host responses linked to Endozoicomonas-mediated coral thermal resilience and offer insight into molecular mechanisms implicated in host-microbe interactions under environmental stress.
Diabetic keratopathy (DK) is a prevalent ocular surface complication of diabetes, frequently unnoticed until significant structural and functional deterioration occurs. Chronic hyperglycemic stress promotes inflammation in the corneal epithelial-neural-immune (epineuroimmune) unit, impeding tissue recovery and increasing infection susceptibility. To address this metabolic-immune-infection imbalance, we developed a light-regulated biomimetic catalytic platform (WCNx-Rh2) that integrates glucose degradation and monitoring, immune-modulated epithelial-neural regeneration, and antibacterial defense. This platform features an S-scheme heterojunction (WCNx) composed of graphitic carbon nitride and tungsten oxide, enabling visible-light (VIS)-driven glucose degradation, dark-state colorimetric detection, and near-infrared (NIR) photothermal antibacterial activity. Complementarily, the loaded ginsenoside Rh2-identified via systematic screening-inhibits the Nucleotide-binding and oligomerization domain (NOD)-like receptor signaling pathway. The combined action of WCNx and Rh2 reduces advanced glycation end products (AGEs), reactive oxygen species (ROS), and inflammatory signaling, reprogramming dendritic cell function to restore epineuroimmune homeostasis and drive tissue repair. These functions were validated in two animal models. In diabetic mice, VIS-irradiated WCNx-Rh2 accelerated corneal epithelial and nerve recovery. In a diabetic keratitis model, NIR-activated WCNx-Rh2 enabled effective corneal bacterial eradication and tissue repair. Overall, this work establishes a light-driven metabolic-modulation biomimetic paradigm and proposes an integrated strategy for managing DK.
Milk oligosaccharides (MO) support intestinal, microbial, and immune development in young pigs. However, modern production practices wean pigs at an early age, removing them from their source of MO prior to intestinal and immune maturation. The purpose of this study was to investigate the effects of dietary supplementation of galacto-oligosaccharides (GOS) and 2'-fucosyllactose (FL) on the jejunal mucosa-associated microbiota, intestinal immune signaling, morphology, and growth performance of nursery pigs. Forty-eight pigs (6.8 ± 0.2 kg body weight) weaned at 3-weeks-of-age were allotted into 6 dietary treatments, using a randomized complete block design, with sex and initial body weight as blocks. Dietary treatments were 1) basal diet; 2) basal diet, with supplemental GOS at 1.5% of the diet; 3) basal diet, with supplemental FL at 0.2% of the diet; 4) basal diet, with GOS and FL at 1.5 and 0.2% of the diet, respectively; 5) basal diet, with GOS at 2.3% of the diet; and 6) basal diet, with FL at 0.3% of the diet. These MO were provided alone or in combination at levels mimicking intake at the end of the suckling period, and at 1.5-fold higher, to observe potential dose-dependent responses. Pigs were fed for 21 d in 2 phases. On d 21, pigs were euthanized for sampling of jejunal mucosa and jejunal tissue. Data were analyzed using the PROC MIXED of SAS 9.4 and contrasts were used to determine the effects of GOS, FL, and their combination (interaction), in addition to the linear effects of increasing dietary GOS or FL. Increasing levels of GOS and FL linearly decreased (P < 0.05) Shannon and Simpson alpha diversity of the jejunal mucosa-associated microbiota. Supplementation with FL increased (P < 0.05) the absolute abundance of Helicobacter in the jejunal mucosa-associated microbiota, although no dose response was observed. Increasing levels of GOS and FL tended to linearly decrease (P = 0.051 and 0.076, respectively) the gene expression of TLR4. Increasing levels of GOS tended to increase (P = 0.054) and increasing supplementation of FL increased (P < 0.05) the number of Ki-67 proliferative cells in the crypt of the jejunum. Increasing levels of GOS increased (P < 0.05) body weight and average daily gain in the early post-weaning period and tended to increase (P = 0.093) body weight by the end of the experimental period. Notably, increasing levels of GOS, and GOS in combination with FL, improved growth performance, whereas FL alone did not. Milk oligosaccharides are indigestible carbohydrates with prebiotic effects for young mammals. Prebiotics support the intestinal microbiota and development of a functional intestine and intestinal immune system. Nursery pigs are weaned at a young age and could benefit from the continued provision of milk oligosaccharides beyond the suckling period to mitigate the negative impacts of weaning on intestinal health. The purpose of this study was to evaluate the effects of two milk oligosaccharides abundantly present in porcine milk, galacto-oligosaccharides and 2’-fucosyllactose, on the jejunal mucosa-associated microbiota, immune response, stress status, morphology, and growth performance of nursery pigs. These oligosaccharides were provided alone or in combination at physiologically relevant levels mimicking intake at the end of the suckling period, and at 1.5-fold higher, to observe potential dose-dependent responses. The use of these milk oligosaccharides modestly influenced the mucosa-associated microbiota, supplementation with galacto-oligosaccharides or 2’fucosyllactose reduced the expression of genes associated with inflammatory signaling and increased intestinal stem cell proliferation. Increasing supplementation of galacto-oligosaccharides up to 1.5 times the physiologically relevant levels improved growth performance of nursery pigs. The combination of both milk oligosaccharides at physiologically relevant levels provided the most consistent benefits related to intestinal function and growth performance of nursery pigs.
Osteoarthritis (OA) is a multifactorial degenerative joint disorder characterized by intricate interactions among oxidative stress, inflammation, apoptosis, and extracellular matrix (ECM) degradation, ultimately disrupting chondrocyte homeostasis and accelerating cartilage deterioration. Although advanced intra-articular biomaterial systems have improved local therapeutic delivery for OA, the rational integration of a multi-component bioactive core with a multifunctional carrier remains challenging. In this study, a multifunctional nanomedicine, K@BMT@HM, was developed by integrating a supramolecular bioactive core with a functionalized hydrogel microsphere carrier. The K@BMT core, composed of the KRFK peptide, bisdemethoxycurcumin (BDMC), and MnTBAP, exerted multi-dimensional regulatory effects by enhancing antioxidant defense, restoring autophagic flux, and modulating TGF-β-associated signaling, thereby mitigating oxidative stress, inflammation, and apoptosis while favoring ECM anabolic balance in chondrocyte-based assays. However, because TGF-β signaling is highly context- and compartment-dependent, these observations should not be interpreted as evidence that TGF-β activation is uniformly beneficial across the whole joint. Meanwhile, the HM-SCHW carrier, composed of chitosan (CS), sodium alginate (SA), hyaluronic acid (HA), and WYRGRL peptide, provided thermosensitive depot formation, lubrication, cartilage-associated localization/retention, and in vitro sustained/ROS-responsive release, thereby supporting localized delivery potential rather than proving WYRGRL-specific cartilage targeting. Moreover, the laser-responsive thermal behavior of K@BMT@HM provided a controllable local heating modality that may support thermosensitive in situ retention. Remarkably, the integrated K@BMT@HM, which couples the multi-component regulatory capacity of the K@BMT core with the multifunctional delivery advantages of the HM-SCHW carrier, was validated through a series of in vitro and in vivo experiments. Collectively, this work presents a rationally designed nanomedicine that integrates bioactive regulation with functionalized delivery to enhance chondroprotection, supporting further preclinical investigation of sustained and multimodal OA intervention. However, because the laser-assisted thermal component was validated only in a rat small-joint model, these findings cannot be directly extrapolated to human OA joints, where deeper articular cartilage and overlying skin, adipose tissue, synovium, and other tissues may substantially limit optical penetration and thermal delivery. In addition, because systematic single-component, two-component, and component-deletion controls were not performed, the present data should be interpreted as validating the integrated K@BMT@HM system as a whole rather than defining the quantitative contribution or indispensability of BDMC, MnTBAP, KRFK, 808-nm laser activation, the CS/SA hydrogel matrix, HA, or WYRGRL.
Pancreatic ductal adenocarcinoma (PDAC) is the most frequent type of pancreatic cancer with a poor prognosis and resistance to conventional radio- and chemotherapy. Radiation can induce pro-inflammatory signaling by activating the type 1 interferon (IFN-I) response, which can be enhanced by targeting negative regulators of the IFN-I pathway, such as ADP-ribosyltransferase PARP7. Here, we show that PARP7 inhibitors (PARP7i) enhance radiation-induced cell death and promote STING- and NF-κB-dependent immunogenic signaling in PDAC cells, leading to inflammatory gene expression and cytokine release. These effects were most pronounced in BxPC-3 cells, which exhibit higher baseline expression of PARP7, AHR, and interferon response genes. PARP7i potentiated the immunogenic effects of hypofractionated radiation by inducing a STING-dependent IFN-I response, leading to immunogenic cell death and activation of monocytes and NK cells. Carbon ion irradiation elicited stronger immunogenicity than X-rays when combined with PARP7i. KRAS-mutated PANC-1 cells showed a higher expression of enzymes that convert ATP to immunosuppressive adenosine, which was enhanced by radiation. This may explain why PARP7i were more effective as monotherapy in PANC-1 cells, promoting NK cell activation. These findings support further evaluation of PARP7i in PDAC in combination with radiotherapy or as monotherapy, depending on the immunosuppressive effects of radiation.