Search Results (12,065)

Increasing evidence suggests that intratumoral microorganisms and their metabolites can modulate cancer initiation and progression. However, the composition and functional role of intratumoral bacteria in canine mammary tumors (CMTs) remain unclear. In this study, we investigated the functional significance of tumor-derived Staphylococcus in CMTs, focusing on its effects on the proliferation and migration of CMT-U27 cells. 16S rRNA sequencing revealed reduced alpha diversity in CMT tissues, with Staphylococcus pseudintermedius identified as the most frequently isolated species. Functional assays, including CCK-8, wound healing, RT-qPCR, and Western blot analyses, demonstrated that intratumoral Staphylococcus pseudintermedius significantly enhanced cellular proliferation and migration. Mechanistically, Staphylococcus pseudintermedius significantly upregulated the expression of TLR2, as well as the phosphorylation levels of PI3K, Akt and P70S6K. The inhibition of TLR2 using C29 suppressed the mRNA expression of VEGF, MMP9, MMP2, and EGFR. Collectively, these findings indicate that intratumoral Staphylococcus pseudintermedius promotes the proliferation and migration of CMT-U27 cells through activation of the TLR2/PI3K/Akt pathway, highlighting a functional link between tumor-associated bacteria and cancer progression. Full article

Etrasimod is an oral selective sphingosine-1 phosphate receptor modulator, and its anti-inflammatory mechanism of action in inflammatory bowel diseases is not completely understood. It targets pro-inflammatory immune cells expressing sphingosine-1-phosphate receptors during their migration from the lymphatic system to the circulation and intestinal mucosa. Reductions in certain lymphocyte subsets in the peripheral blood have been reported, but its effects in lymph nodes remain unknown. This study investigated changes in leukocyte subpopulations in peripheral lymph nodes and blood in Crohn’s disease patients treated with etrasimod. Moderate-to-severe Crohn’s disease patients participated in this randomized, double-blind study, within the phase 2 CULTIVATE clinical trial. At baseline and after 14 weeks of etrasimod treatment, peripheral blood and inguinal lymph node biopsies were obtained. Isolated peripheral blood mononuclear cells and lymph node leukocyte populations were analyzed at single cell level using mass cytometry at both timepoints. The immunophenotyping revealed 15 innate and adaptive major immune cell populations, as well as 14 subpopulations of CD4+ and CD8+ T-cells. In peripheral lymph nodes, etrasimod resulted in significant accumulation of naïve, central memory, and effector memory CD4+ T-cells (+10.7%, +4.2%, and +2.3%, respectively; all p = 0.03), as well as naïve CD8+ T-cells (+4.2%; p = 0.03). Conversely, these subsets were reduced in peripheral blood (−6.2%, −6.0%, −2.0%, and −2.2%, respectively; all p = 0.03). Naïve and memory B-cells decreased in the circulation (−1.7%, p = 0.057; −0.6%, p = 0.03, respectively) but were unchanged in the lymph nodes. Innate immune cell populations remained mostly unaffected in both compartments. Our data indicate that etrasimod’s pharmacodynamic effect is related primarily with the attenuation of the T-cell mediated inflammation with minor changes in B-cells. However, additional follow-up studies are needed for the validation of these observations in the context of Crohn’s disease. Full article

Melanoma is an aggressive cancer characterized by a rapid metastatic process. Thus, understanding the mechanisms underlying its progression is urgently needed to improve patient outcomes. In this regard, there is consistent evidence of a tumor-sustaining crosstalk between melanoma and subcutaneous adipose tissue; however, the role of extracellular vesicles (EVs) in this communication still needs to be clarified. We demonstrated that the EVs derived from adipocytes did not alter melanoma cell proliferation but significantly promoted tumor cell migration and invasion by determining an enrichment in mesenchymal markers, such as N-cadherin and vimentin. In particular, these changes were accompanied by the transition towards a stem-like phenotype, characterized by enhanced spherogenic ability and ABCG2 upregulation; interestingly, this led to a reduced in vitro response to the BRAF inhibitor vemurafenib. Mechanistically, an increase in PGC-1α expression was found, resulting in higher mitochondrial mass and activity, ATP synthesis, and ROS overproduction; of note, treatment of melanoma cells with SR-18292 and XCT790, two inactivators of mitochondrial biogenesis, and N-acetylcysteine, a ROS scavenger, successfully counteracted the above EV-related effects, suggesting that mitochondrial function could be targeted to suppress the vesicular interactions between adipose tissue and melanoma. Taken together, these results highlight the crucial role played by EVs in melanoma stroma, pointing out the ability of adipocyte-derived vesicles to sustain cancer aggressiveness via PGC-1α–dependent mitochondrial reprogramming. Full article

Cortex Lycii Radicis, a medicinal plant, has been reported to inhibit epithelial–mesenchymal transition (EMT) and exhibit anti-lung cancer properties. Our previous study identified its major compound, Kukoamine B (KuB), as an inhibitor of membrane PD-1/PD-L1 interaction, thereby restoring T-cell function. However, the effect of KuB on EMT and the underlying mechanism thereof remain unknown. Herein, we show that PD-L1 overexpression enhances the proliferation, migration, and EMT of LUAD cells, upregulating N-cadherin and Vimentin, while downregulating E-cadherin. Mechanistically, PD-L1 directly binds phosphorylated p65 (p-p65) and facilitates p65 nuclear translocation, an interaction confirmed by molecular simulations. We found that KuB disrupts the PD-L1/p65 complex, impedes p65 nuclear translocation, and suppresses EMT, proliferation, and migration in LUAD cells. These inhibitory effects were reversed by PD-L1 overexpression. We therefore conclude that KuB suppresses EMT in LUAD by targeting intracellular PD-L1, blocking PD-L1–p65 interaction and nuclear translocation of p65. Full article

Endothelial cells are key regulators of vascular homeostasis, and their dysfunction plays a central role in the development of atherosclerosis and other cardiovascular diseases. Multinucleated variant endothelial cells (MVECs) have been described in pathological vascular regions; however, their functional properties remain poorly characterized. The aim of the present study was to compare lipid handling, inflammatory activation, barrier-associated features, and secretory profiles of typical endothelial cells (TECs, EA.hy926 line) and MVECs under low-density lipoprotein (LDL) exposure. MVECs were generated by polyethylene glycol-induced fusion of EA.hy926 cells and incubated with LDL under standardized conditions. Intracellular cholesterol accumulation was assessed biochemically, cytokine secretion was quantified by ELISA, gene expression of inflammatory, endothelial, junctional, and vasoactive markers was analyzed by quantitative real-time PCR, and the endothelial secretome was characterized using data-independent acquisition liquid chromatography–tandem mass spectrometry (DIA-LC-MS). MVECs demonstrated enhanced cholesterol accumulation compared with TECs following LDL exposure. At the transcriptional level, MVECs were characterized by elevated basal expression of proinflammatory markers, including IL1B, IL6, and NFKB1, and showed a markedly amplified IL6 and IL8 response to LDL. In parallel, MVECs exhibited reduced expression of genes associated with antioxidant defense (SOD1), barrier integrity (TJP1), and hemostatic function (VWF). Consistent with transcriptional data, mass spectrometry-based secretome analysis revealed decreased secretion of von Willebrand factor (vWF), vascular endothelial growth factor C (VEGFC), and endothelin-1 (EDN1) by MVECs, accompanied by increased secretion of tissue-type plasminogen activator (t-PA). Functional enrichment analysis of secretome-associated proteins highlighted pathways related to extracellular matrix–receptor interaction, focal adhesion, cell adhesion molecules, complement and coagulation cascades, and leukocyte transendothelial migration. In contrast, TECs demonstrated a more pronounced transcriptional response in EDN1, consistent with their role in vascular tone regulation. Immunocytochemical analysis further revealed altered subcellular distribution of the tight junction protein ZO-1 in MVECs, indicating junctional destabilization. Taken together, these results indicate that MVECs represent a distinct endothelial phenotype characterized by enhanced lipid accumulation, sustained proinflammatory activation, altered secretory signaling, and reduced barrier and hemostatic potential. Such features suggest that MVECs may contribute to the maintenance of chronic endothelial dysfunction and vascular inflammation under conditions of lipid overload. Full article

Background: Lung cancer remains a leading cause of mortality, mainly due to aggressive metastasis and therapeutic resistance. Snake venom metalloproteinases (svMPs), particularly the P-I class, are promising sources for novel antitumor agents. Objectives: This study investigated the impacts of BthMP, a P-I svMPs from Bothrops moojeni venom, on human lung carcinoma (A549) cells in comparison to non-cancerous human bronchial epithelial cells (BEAS-2B). Methods and Results: BthMP demonstrated potent and selective anti-cancer activity. It significantly inhibited key metastatic processes in A549 cells, including adhesion, migration, and invasion, while suppressing long-term proliferation, as shown by reduced colony formation and increased lactate dehydrogenase (LDH) release. Mechanistically, BthMP induced a massive increase in intracellular reactive oxygen species (ROS) by over 2000% and elevated nitric oxide (NO) by 35% in A549 cells, driving a state of lethal oxidative stress. Crucially, these cytotoxic and anti-metastatic effects were minimal in BEAS-2B cells; BthMP even suppressed basal ROS and NO levels in this non-cancerous line. The anti-migratory effects of BthMP were completely dependent on its zinc-based catalytic activity, as they were abolished by pretreatment with ethylenediaminetetraacetic acid. By simultaneously disrupting cell–matrix interactions and inducing selective, catastrophic oxidative stress in cancer cells, BthMP presents a dual-pronged anti-metastatic mechanism. Conclusions: These findings establish BthMP as a promising therapeutic scaffold for developing novel treatments against lung cancer progression. Full article

The blood–brain barrier (BBB), a core component of the neurovascular unit (NVU), meticulously regulates material exchange between the blood and brain parenchyma, serving as a critical barrier for maintaining the homeostasis of the central nervous system (CNS). Neuroinflammation, a pivotal response of the CNS to injury and disease, can disrupt NVU homeostasis when excessive or persistent, acting as a core pathogenic driver of various intractable neurological disorders. Chemokines, as key signaling molecules guiding the directional migration of immune cells, form the central hub mediating the dynamic regulation of neuroinflammation and the BBB. However, existing studies mostly focus on single disease systems or chemokine families, neglecting the bidirectional heterogeneity of different chemokine axes in BBB regulation and the common regulatory rules across diseases, while lacking systematic exploration of clinical translation challenges caused by the redundancy and spatiotemporal heterogeneity of the chemokine network. This review systematically clarifies the bidirectional regulatory effects of the core axes of the three major chemokine families (e.g., CCL2/CCR2, CXCL12/CXCR4, CX3CL1/CX3CR1) on the BBB. For the first time, we integrate a multi-dimensional regulatory model based on concentration, location, and time to analyze their molecular mechanisms and regulatory heterogeneity in promoting BBB disruption under pathological conditions versus mediating barrier repair and neuroprotection under specific spatiotemporal conditions. Combined with advancements in cutting-edge models such as microfluidic chips, we discuss the clinical translation progress of chemokine research, including potential biomarkers and targeted therapeutic strategies, and propose precise breakthrough paths for the two core challenges of network redundancy and spatiotemporal heterogeneity. Finally, we construct a complete research framework for chemokine-mediated regulation of NVU homeostasis, providing novel insights and directions for restoring BBB function and treating intractable neurological diseases. Full article

Osteosarcoma, the most common malignant bone tumor in young individuals, often exhibits poor outcomes due to MDM2-mediated suppression of the p53 pathway. Whereas conventional MDM2 inhibitors block the p53–MDM2 interaction but frequently induce compensatory MDM2 upregulation, proteolysis-targeting chimeras (PROTACs) directly degrade MDM2 and bypass this limitation. Here, we investigated the anticancer efficacy of two MDM2-targeting PROTAC compounds, CL0144 and CL0174, in osteosarcoma models. In Saos-2 and U2OS cells, both PROTACs efficiently induced MDM2 degradation, leading to activation of p53 or p73 signaling, increased reactive oxygen species production, apoptotic cell death, and marked reductions in viability. PROTAC treatment also significantly suppressed proliferation, colony formation, sphere formation, migration, and invasion. In vivo, xenograft assays demonstrated robust tumor growth inhibition following PROTAC administration. Collectively, these findings demonstrate that MDM2-targeting PROTACs exert strong antitumor effects by degrading MDM2 and disrupting downstream oncogenic pathways, supporting their potential as a promising therapeutic strategy for osteosarcoma. Full article

Background: Artemisia annua L. is a medicinal plant with documented antimicrobial, antioxidant, and anti-inflammatory properties. Although widely studied for internal therapeutic applications, its topical use—especially in hydrogel-based systems—has not been thoroughly investigated. The aim of this study was to develop sodium alginate hydrogels containing Artemisia annua extract, supplemented with hyaluronic acid and dexpanthenol, and to evaluate their physicochemical characteristics as well as their biological activities in vitro and in vivo. Methods: Select bioactive constituents of the Artemisia annua extract were quantified using liquid chromatography coupled with electrospray ionization mass spectrometry (LC-ESI-MS). Hydrogels were prepared by cross-linking sodium alginate with a calcium carbonate–glucono-delta-lactone system and were formulated with or without hyaluronic acid and dexpanthenol. Physicochemical evaluations included measurements of moisture content, water-retention capacity, gelation time, and pH. The hydrogel microstructure was examined by scanning electron microscopy (SEM). Antioxidant activity was assessed using three methods: the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, the ferric reducing antioxidant power (FRAP) assay, and the cupric reducing antioxidant capacity (CUPRAC) assay. Biocompatibility and regenerative effects were analyzed using cell viability assays and an in vitro scratch wound model on human keratinocyte cells. In vivo wound-healing efficacy was examined in rats with full-thickness skin excisions. Results: The extract contained high levels of methylated flavonoids and sesquiterpenes characteristic of Artemisia annua. Hydrogels supplemented with hyaluronic acid and dexpanthenol exhibited improved hydration stability and higher porosity. All formulations demonstrated measurable antioxidant activity, and those containing hyaluronic acid showed the strongest effects. The preparations were biocompatible and enhanced keratinocyte migration in vitro, with the combined hyaluronic acid–dexpanthenol formulation promoting the fastest wound closure. In vivo, Artemisia annua hydrogels accelerated wound healing by two to three days compared with untreated wounds. Conclusions: These results confirm the promise of Artemisia annua hydrogels for topical wound care and highlight the beneficial contributions of hyaluronic acid and dexpanthenol to their structural and therapeutic performance. Full article

Current wound healing strategies must confront numerous challenges. Helminth-induced immunomodulation offers a promising therapeutic avenue for inflammatory diseases and injury repair. However, research on the role of helminths in damage recovery remains limited. We utilized glycogen—a naturally occurring biomaterial—to encapsulate SJMHE1, a bioactive peptide derived from Schistosoma japonicum, and successfully developed a facilely prepared hydrogel formulation denoted as SJMHE1-gel. The properties of SJMHE1-gel, its effect on cell activity, and its performance in a murine full-thickness skin defect model were evaluated. The glycogen-based hydrogel exhibited a uniform pore size, excellent biocompatibility, and sustained release of SJMHE1. Topical application of SJMHE1-gel enhanced collagen deposition, promoted angiogenesis, facilitated the regeneration of hair follicles and sebaceous glands, and accelerated full-thickness wound healing. SJMHE1-gel also promoted M2 macrophage polarisation and suppressed inflammatory cytokine expression both in vivo and in vitro. Mechanistically, SJMHE1-treated macrophages upregulate TGF-β, which in turn promotes the migration of L929 fibroblasts and human umbilical vein endothelial cells (HUVECs) via the Smad3 pathway. Neutralization of TGF-β attenuates phosphorylated Smad3 (p-Smad3) levels and impairs the migratory capacity of both fibroblasts and HUVECs. Additionally, SJMHE1-treated macrophages upregulate VEGFA, thereby enhancing angiogenic tube formation in HUVECs. This easy-to-prepare hydrogel can regulate macrophage polarization, inhibit inflammation, promote angiogenesis, and accelerate collagen deposition, acting across wound healing stages to provide a novel therapeutic strategy. Full article

Pulp necrosis remains a significant clinical challenge in dentistry, as current therapeutic approaches fail to achieve functional pulp regeneration. Extracellular vesicles (EVs), as crucial mediators of intercellular communication, offer new opportunities for regenerative strategies. In this study, we focus on CD24⁺ human dental papilla cells (CD24⁺ hDPCs), a functionally defined subpopulation previously characterized as having superior regenerative potential, and evaluate the regenerative potential of their derived EVs (CD24⁺ EVs) in pulp-like tissue regeneration. CD24⁺ EVs significantly enhanced the proliferation, migration, and osteo/odontogenic differentiation of human dental pulp stem cells (hDPSCs) and markedly promoted endothelial tube formation in vitro. In a treated dentin matrix (TDM)-based ectopic regeneration model, CD24⁺ EVs increased cellular accumulation within the regenerated tissue and robust angiogenesis, inducing the formation of well-organized, highly vascularized pulp-like tissue with dense cellular architecture and positive DSPP expression. Together, these findings suggest that CD24⁺ EVs concurrently enhance cell migration, odontogenic differentiation, and angiogenesis, and support a promising cell-assisted EV strategy grounded in functionally defined cellular subpopulations for pulp-like tissue regeneration. Full article

Breast cancer is the most common malignant tumor among women worldwide, and its occurrence is closely associated with long-term exposure to environmental pollutants. Polycyclic aromatic hydrocarbons (PAHs) are a class of persistent organic pollutants widely present in the living environment. Epidemiological studies indicate that exposure to PAHs increases the risk of breast cancer. PAH derivatives exhibit stronger toxicity or endocrine-disrupting activity than their parent compounds; however, research on their specific effects and mechanisms in breast cancer cells remains limited. For this purpose, this study selected 3-Hydroxybenz[a]anthracene, a PAH derivative with potential estrogenic activity, as the target compound. Using the estrogen receptor-positive breast cancer cell line MCF-7 as the model, we performed EdU staining, colony formation assays, scratch healing assays, Transwell invasion assays, and apoptosis assays and preliminarily examined changes in relevant signaling proteins via Western blot. Results indicate that 3-Hydroxybenz[a]anthracene promotes proliferation and migration in MCF-7 cells while inhibiting apoptosis under certain conditions, but it has no effect on cell invasion. Mechanistically, it upregulates key proteins including AKT, c-Myc, E-Cadherin, Vimentin, MMP2, MMP9 and Bcl-2 while downregulating p-AKT expression. This study confirms through in vitro experiments that 3-Hydroxybenz[a]anthracene exhibits estrogen-like effects and modulates malignant behavior in breast cancer cells by regulating relevant signaling pathways. These findings provide experimental evidence for further evaluating the potential role of this environmental contaminant in breast cancer initiation and progression. Full article

Pulmonary arterial hypertension (PAH) is a progressive disease characterized by endothelial dysfunction, vascular remodeling, and a sustained increase in pulmonary vascular resistance, causing cardiopulmonary damage. The apelin receptor (APJ), a member of the G protein-coupled receptor family, has emerged as an essential modulator of vascular homeostasis. Clinical and preclinical studies have demonstrated that its activation exerts beneficial effects on the progression of PAH. Its main actions include the restoration of endothelial function, reactivation of the BMPR2/SMAD axis, induction of nitric oxide-mediated vasodilation, inhibition of autophagy and the migration of the pulmonary artery smooth muscle cells (PASMCs). Furthermore, its expression and functionality are modulated by epitranscriptomic mechanisms, particularly by microRNAs involved in the post-transcriptional regulation of key genes for vascular homeostasis. These findings position the APJ as a relevant therapeutic target in PAH. However, the clinical application of its agonists still faces pharmacokinetic limitations that restrict their therapeutic use. Therefore, the aim of this review is to gather current information on APJ in the pathophysiology of PAH and focus attention on its potential as a therapeutic target. Full article

The red resin of Dracaena cochinchinensis (Lour.) S.C. Chen, known as Chinese dragon’s blood, is formed through metabolic reprogramming following trunk injury, during which the original steroidal saponins are depleted and transformed. To investigate the steroidal degradation intermediates in this process, a systematic phytochemical study was conducted on the resin from Yunnan Province, leading to the isolation of 14 steroidal constituents (2 new and 12 known). The two new compounds, dracaenogenins C (1) and D (2), were identified as rare 12(13→14)-abeo-spirostanol aglycones, with 2 representing an unusual C-14α-hydroxylated derivative. Their structures, including absolute configurations, were unambiguously determined by comprehensive spectroscopic analysis (1D and 2D NMR, HRESIMS) and single-crystal X-ray diffraction. Biogenetic analysis suggests that these unusual aglycones arise from the acid-catalyzed Wagner–Meerwein rearrangement of diosgenin-type saponins via C-18 angular methyl migration (C-10→C-13) and C-ring contraction, serving as rare catabolic intermediates trapped during the metabolic shift from saponin accumulation to polyphenol biosynthesis. Furthermore, cytotoxicity evaluation against HepG2 cells revealed that while the parent glycosylated saponins (e.g., dioscin and gracillin) exhibited significant toxicity, the rearranged aglycones (1, 2, and 3) and other degradation products were devoid of cytotoxicity, supporting a detoxification mechanism during resin formation. Full article

Prostate cancer (PCa) is the most general cancer in men and is often linked with distant metastasis in its later stages. The caffeic acid (CA) derivative, N-(4-methoxyphenyl)methylcaffeamide (MPMCA), demonstrates superior liver-protective effects compared to CA. Nevertheless, the functions of MPMCA on prostate cancer metastasis remain unclear. Here, we demonstrate that MPMCA blocks migration and invasion in prostate cancer cells without affecting cell viability. By suppressing the production of mesenchymal markers Vimentin, N-cadherin and β-catenin and upregulating the production of the epithelial marker Zonula Occludens-1 (ZO-1), MPMCA also controls Epithelial–Mesenchymal Transition (EMT). The Phosphoinositide 3-kinase (PI3K), Protein kinase B (AKT) and mechanistic target of rapamycin (mTOR) pathway has been documented to regulate MPMCA-inhibited cell motility. Transfection with Snail and Slug cDNA reverses MPMCA’s suppression of EMT, migration, and invasion in prostate cancer cells. Importantly, our in vivo data indicates that MPMCA reduces Snail and Slug expression and prostate cancer metastasis. Our evidence suggests that MPMCA is a novel therapeutic candidate for treating metastatic prostate cancer. Full article