Search Results (176)

Background: Anti-programmed death 1 receptor (PD-1) therapy is a promising treatment strategy for patients with unresectable advanced or recurrent gastric/gastroesophageal junction (G/GEJ) cancer. However, its response rate and survival benefits are still limited; an immunological analysis of the residual tumor after anti-PD-1 therapy would be important. Methods: We evaluated the clinical efficacy of tumor resection (TR) after chemotherapy or anti-PD-1 therapy in patients with unresectable advanced or recurrent G/GEJ cancer and analyzed the immune status of tumor microenvironment (TME) by immunohistochemistry using their surgically resected specimens. Results: Patients treated with TR after anti-PD-1 therapy had significantly longer survival compared to those treated with chemotherapy and anti-PD-1 therapy alone. Expression of human leukocyte antigen (HLA) class I and major histocompatibility complex (MHC) class II on tumor cells was markedly downregulated after anti-PD-1 therapy compared to chemotherapy. Furthermore, the downregulation of HLA class I may be associated with the activation of transforming growth factor-β signaling pathway in the TME. Conclusions: Immune escape from cytotoxic T lymphocytes may be induced in the TME in patients with unresectable advanced or recurrent G/GEJ cancer after anti-PD-1 therapy due to the downregulation of HLA class I and MHC class II expression on tumor cells. TR may be a promising treatment strategy for these patients when TR is feasible after anti-PD-1 therapy. Full article

Myocardial infarction (MI) remains one of the most common cardiovascular diseases and a leading cause of morbidity and mortality worldwide. In recent years, natural polymeric patches have attracted increasing attention as a promising therapeutic platform for myocardial tissue repair. This study explored the fabrication and evaluation of screen-printed electrodes (SPEs) on chitosan film as a novel platform for cardiac patch applications. Chitosan is a biodegradable and biocompatible natural polymer that provides an ideal substrate for SPEs, providing mechanical stability and promoting cell adhesion. Silver ink was employed to enhance electrochemical performance, and the electrodes exhibited strong adhesion and structural integrity under wet conditions. Mechanical testing and swelling ratio analysis were conducted to assess the patch’s physical robustness and aqueous stability. Silver ink was employed to enhance electrochemical performance, which was evaluated using cyclic voltammetry. In vitro, electrical stimulation through the chitosan–SPE patch significantly increased the expression of cardiac-specific genes (GATA-4, β-MHC, troponin I) in bone marrow mesenchymal stem cells (BMSCs), indicating early cardiogenic differentiation potential. In vivo, the implantation of the chitosan–SPE patch in a rat MI model demonstrated good tissue integration, preserved myocardial structure, and enhanced ventricular wall thickness, indicating that the patch has the potential to serve as a functional cardiac scaffold. These findings support the feasibility of screen-printed electrodes fabricated on chitosan film substrates as a cost-effective and scalable platform for cardiac repair, offering a foundation for future applications in cardiac tissue engineering. Full article

Background/Objectives: Contagious agalactia (CA) is a disease typically caused by Mycoplasma agalactiae, affecting small ruminants worldwide and being endemic in certain countries. CA causes severe economic losses due to mastitis, agalactia, and arthritis. As an alternative to existing immunoprophylactic measures, this study aimed to develop a recombinant subunit vaccine against M. agalactiae and evaluate its specific immune response in goats. Methods: Goats were divided into three groups: group 1 received recombinant proteins (P40 and MAG_1560), group 2 received formalin-inactivated M. agalactiae, and group 3 received Tris-buffered saline (negative control). All solutions were emulsified in Freund’s adjuvant. Animals were monitored for 181 days. IgG antibody production was assessed by ELISA, and peripheral blood mononuclear cells (PBMCs) were analyzed by real-time PCR for the expression of IL-1β, IFN-γ, IL-12, and MHC class II genes. Results: M. agalactiae-specific antibody response was observed for six months in the sera of animals from group 1. Analysis of cytokine gene expression revealed increased IL-1β mRNA levels over time in both experimental groups. In group 1, IFN-γ mRNA levels increased with P40 stimulation and decreased with MAG_1560. IL-12 mRNA expression decreased over time in group 1 with P40 stimulation, whereas group 2 showed increased IL-12 expression for both proteins. MHC-II expression was stimulated in both groups. Conclusions: The recombinant proteins induced antibody production and cytokine expression, demonstrating immunogenic potential and supporting their promise as vaccine candidates capable of eliciting both humoral and cellular immune responses against M. agalactiae. Full article

Peripheral nerve injury initiates a complex cascade of events coordinating immune responses, extracellular matrix (ECM) remodeling, and neuronal repair. While β2-microglobulin (B2M) is well known for its role in MHC class I-mediated antigen presentation and CD8⁺ T-cell differentiation, its potential contributions to non-immune processes remain underexplored. In this study, we investigated the role of B2M in peripheral nerve regeneration using a chronic constriction injury (CCI) model in wild-type and B2M-deficient (B2M-KO) mice. Flow cytometry, RNA sequencing (RNA-seq), and quantitative PCR (qPCR) were performed to assess T-cell subset dynamics and gene expression following injury. Flow cytometric analysis showed that CD3⁺CD4⁺ and CD3⁺CD8⁺ T-cell populations increased by day 7 post-injury. While CD3⁺CD4⁺ T-cell expansion occurred in both groups, a significant increase in CD3⁺CD8⁺ T cells was observed only in wild-type mice. RNA-seq analysis at 3 days post-injury—prior to substantial T-cell accumulation—revealed marked downregulation of ECM-related genes in B2M-KO mice, including collagens, matrix-associated proteins, and other key ECM components. KEGG analysis identified suppression of ECM–receptor interaction, PI3K-Akt, and TGF-β signaling pathways. qPCR confirmed reduced expression of Thbs1 in B2M-KO mice. These findings suggest that B2M plays a critical, CD8⁺ T-cell-independent role in regulating ECM dynamics and regenerative signaling during early nerve repair, expanding the conceptual framework of B2M’s function beyond classical immune roles. Full article

Background/Objectives: Myogenesis, the process by which myoblasts differentiate into multinucleated muscle fibers, is tightly regulated by transcription factors, signaling pathways, and metabolic cues. Among these, fatty acids have emerged as key regulators beyond their traditional role as energy substrates. Oleic acid, a monounsaturated fatty acid, has been shown to modulate muscle differentiation, potentially influencing myogenic pathways. This study examines the role of oleic acid in promoting C2C12 myoblast differentiation and its associated molecular mechanisms, comparing it to standard horse serum (HS)-based differentiation protocols. Methods: C2C12 murine myoblasts were cultured under proliferative conditions and differentiated using DMEM supplemented with either 2% HS or oleic acid (C18:1, n-9). The molecular signaling pathway was evaluated by measuring the expression of p38 MAPK, β-catenin, GLUT4, and NDRG1. Results: Oleic acid promoted the differentiation of C2C12 cells, as evidenced by a progressively elongated morphology, as well as the induction of muscle-specific myogenin, myosin heavy chain (MHC), and MyoD. Moreover, oleic acid reduced the expression of Atrogin-1 and MuRF1 ubiquitin E3 ligase. BODIPY staining revealed the enhanced accumulation of lipid droplets in oleic acid-treated cells. The Western blot analysis demonstrated robust activation of p38 MAPK and β-catenin pathways in response to oleic acid, compared with HS. Additionally, oleic acid upregulated GLUT4 expression and increased the phosphorylation of insulin receptor and NDRG1, indicating an enhanced glucose uptake capacity. Conclusions: These findings demonstrate that oleic acid promotes C2C12 myoblast differentiation and improves glucose uptake via GLUT4. Oleic acid emerges as a promising metabolic regulator of myogenesis, offering potential therapeutic applications for muscle regeneration in muscle-related pathologies. Full article

BAP31, a transmembrane protein in the endoplasmic reticulum, is known for its oncogenic properties, but its role in immunotherapy is not well understood. While BAP31’s involvement in liver, gastric, and cervical cancers has been documented, its role in pan-cancer immune regulation, particularly in breast cancer, remains unexplored. Using TCGA data, analysis via the Xiantao academic and GEPIA2 database showed that BAP31 upregulation correlates with advanced clinical stages and a poor prognosis. ROC analysis demonstrated BAP31’s high accuracy in distinguishing cancerous tissue from normal tissues. Additionally, BAP31 expression is associated with CNV, methylation, TMB, and MSI. In breast cancer, TIMER database analysis revealed that BAP31 expression is inversely correlated with the infiltration levels of myeloid-derived suppressor cells (MDSCs), macrophages, T lymphocytes, B lymphocytes, and neutrophils. Additionally, we investigated the relationship between BAP31 and the expression of major histocompatibility complex (MHC) molecules and chemokine receptors utilizing the TISIDB database. LinkedOmics analysis demonstrated associations between BAP31 and various immune-inflammatory pathways, while also indicating a negative correlation between BAP31 expression and four critical pathways: extracellular matrix receptor interaction, focal adhesion, JAK-STAT signaling, and TGF-β signaling. Furthermore, loss-of-function experiments employing shRNA-mediated knockdown of BAP31 resulted in a marked reduction in cell proliferation and an increase in apoptosis in breast cancer cells, thereby confirming its role in tumor promotion. These findings suggest that BAP31 may serve as a promising prognostic biomarker and a potential target for immunotherapy in breast cancer. Full article

Background: The increasing incidence of infection with Gram-negative bacilli (GNB) producing broad-spectrum β-lactamases, such as extended-spectrum β-lactamases (ESBLs), cephalosporinases (AmpCs), and carbapenemases, has become a great clinical concern. AmpCs are found in many clinically relevant Enterobacterales, where they may compromise the effectiveness of most β-lactams, including carbapenems when associated with an impaired outer membrane. Detection and distinction between these resistance mechanisms are crucial for antimicrobial therapy and for implementation of proper infection control procedures to prevent further spread. Methods: The disk diffusion antibiogram using Mueller-Hinton agar (MHA) supplemented with cloxacillin (MHC), which inhibits AmpCs, was validated to identify AmpC-producing Enterobacterales (AmpC-PE). As cloxacillin is not available in several countries, we investigated the use of oxacillin as an alternative compound to inhibit AmpCs. The ability of MHA supplemented with oxacillin (MHO) to distinguish between carbapenem-resistant Enterobacterales (CREs) due to AmpC hyperproduction and the presence of a carbapenemase has particularly been investigated. Results: MHOs containing several concentrations of oxacillin were compared to MHA and MHC containing 250 mg/L cloxacillin (MHC250). A set of well-characterized Enterobacterales with different β-lactam resistance mechanisms were evaluated. MHO containing 300 mg/L of oxacillin (MHO300) gave similar results to MHC250. Conclusions: The use of MHO300 proved to be efficient in inhibiting AmpCs, allowing differentiation between AmpC hyperproducers and carbapenemase producers. In addition, the use of MHO300 allowed detection of resistance mechanisms hidden by AmpCs, such as ESBLs. Full article

Vitamin D plays a vital role in human physiology, including a crucial role in regulating bone metabolism and various extra-skeletal effects. Calcitriol exerts anti-inflammatory effects on monocytes and macrophages by increasing IL-10 production and decreasing the production of proinflammatory IL-1β, IL-6, tumor necrosis factor-α (TNF-α), receptor activator of nuclear factor kappa-Β ligand (RANKL), and cyclo-oxygenase-2 (COX-2). In addition, calcitriol also exerts important effects on adaptive immunity by downregulating MHC-II class and co-stimulatory molecules on antigen-presenting cells, but it also directly affects T lymphocytes. In multiple studies, the influence of vitamin D on eye diseases, including corneal diseases, has been demonstrated. Adequate vitamin D supplementation in patients with dry eye significantly improves tear quality and consequently heals the epithelial cells of the ocular surface. Pterygium is a fibrovascular growth of conjunctival tissue that histologically consists of superficial conjunctival epithelium and an underlying fibrovascular layer. The prevalence of pterygium depends on the region. In zones closer to the equator—“pterygium zone”, it is up to 22%, and outside of them it can be even less than 2%. While UV radiation is recognized as a primary risk factor, other factors, including vitamin D, may influence its development. This review aims to summarize the effects of vitamin D on the pathophysiological mechanism of pterygium and its possible therapeutic impact. Current research suggests that vitamin D is protective through its immunomodulatory and anti-inflammatory properties. Finally, there is still insufficient evidence of the therapeutic benefit of vitamin D in pterygium, and future large-scale randomized controlled studies are needed to elucidate the exact role of vitamin D in pterygium onset and recurrence as well as its potential therapeutic benefit. Full article

Multipotent vascular stem cells (MVSCs) are found in the vascular wall and surrounding tissues and possess the ability to differentiate into mesenchymal lineages. Previous studies have shown that MVSCs can be activated in response to vascular injury and differentiate into vascular smooth muscle cells (SMCs), contributing to vascular remodeling and microvessel formation. However, it remains unclear as to whether and how microenvironmental changes in the extracellular matrix, such as substrate stiffness, modulates MVSC differentiation under pathological conditions. This study demonstrated that MVSCs cultured on stiff substrates exhibited increased cell spreading, stronger cell adhesion, and a higher expression of SMC markers, including myosin heavy chain (MHC), myocardin (MYCD), calponin 1 (CNN1), and smooth muscle α-actin (SMA). In contrast, MVSCs on soft substrates showed an elevated expression of the chondrogenic markers aggrecan 1 (AGC1) and collagen-II (COL2A1). The presence of TGF-β1 further increased the expression of SMC markers on stiff substrates and chondrogenic markers on the soft substrates. Collectively, these results establish substrate stiffness as a key regulator of MVSC lineage commitment through cytoskeletal reorganization, with TGF-β1 acting as a biochemical amplifier. Our findings highlight the substrate-stiffness-dependent differentiation of MVSCs and provide mechanistic insights into the role of MVSCs in vascular remodeling during atherosclerosis development and blood vessel regeneration. Full article

Introduction. The pathophysiology of Pyoderma Gangrenosum (PG) involves altered innate and adaptive immunity, mutagenic and epigenetic changes, the autoinflammatory state, and the overexpression of cytokines. This study investigated the potential contribution of inflammation, redox signaling, and the immune system in the pathogenesis of PG. Materials and Methods. This case–control study included 36 patients with PG and 30 controls. We have determined the serum concentrations of acute phase proteins (C-reactive protein—CRP, alpha1 glycoprotein acid—AGPA, Albumin), interleukin-17A -IL-17A, β2 microglobulin-β2MG, reduced glutathione-GSH, oxidized glutathione- GSSG, the GSH/GSSG ratio, and hematological parameters (white blood cells-WBC, neutrophil-lymphocyte ratio-NLR, erythrocyte sedimentation rate-ESR) in patients with PG compared with controls. Furthermore, we have evaluated the variations in these markers before and after treatment in PG patients. Results. The serum concentrations of acute phase proteins (CRP, AGPA, and Albumin) and the IL-17A, β2MG, GSH, GSSG, and GSH/GSSG ratio were significantly different between the PG group and controls. Hematological parameters (WBC, NLR, and ESR), acute phase proteins (CRP, AGPA, and albumin), and IL-17A showed an exaggerated and persistent inflammatory response in patients with PG. In patients with PG associated with systemic diseases, the dysregulation of the biochemical events was more severe. Conclusions. The acute phase proteins, β2MG-MHC class I complex, and the GSH-GSSG system are unbalanced in PG. Our results could improve the diagnosis and our understanding of the pathogenic basis of PG. Full article

Growth differentiation factor 6 (GDF6), a member of the TGF-β superfamily, plays multifaceted roles in tumorigenesis, yet its molecular mechanisms and cancer-type-specific regulatory networks remain poorly defined. This study investigates GDF6’s context-dependent functions through pan-cancer multi-omics integration and functional validation. Transcriptomic data from TCGA (33 cancers, n = 10,535) and GTEx were analyzed to assess GDF6 dysregulation. Co-expression networks, pathway enrichment (KEGG/GO), and epigenetic interactions (m6A, m5C, m1A) were explored. Functional assays included siRNA knockdown, wound healing, and validation in immunotherapy cohorts. GDF6 exhibited bidirectional expression patterns, with downregulation in 23 cancers (e.g., GBM, BRCA) and upregulation in 7 malignancies (e.g., KIRC, PAAD). Mechanistically, GDF6 activated the PI3K-Akt/VEGF pathways, thereby promoting angiogenesis and metastasis. It modulated epigenetic regulation through interactions with m6A readers and erasers. Additionally, GDF6 reshaped the immune microenvironment by recruiting myeloid-derived suppressor cells (MDSCs) and cancer-associated fibroblasts. Notably, GDF6’s dual role extended to immunotherapy: it suppressed anti-PD1 efficacy but enhanced anti-PD-L1 sensitivity, linked to differential MHC-II and hypoxia-response regulation. This study deciphers GDF6’s context-dependent molecular networks, revealing its dual roles in metastasis and immune evasion. These findings highlight GDF6 as a central node in TGF-β-mediated oncogenic signaling and a potential therapeutic target for precision intervention. Full article

Polyphenol-rich plant products are widely used as feed additives for their anti-inflammatory, antioxidant, and antimicrobial properties. The aim of this research is to test the hypothesis that feeding grape pomace (GP) to fattening pigs modulates the intestinal immune and antioxidant response, promotes the morphostructure of the small intestine, and improves growth performance. Fifty Piétrain fattening pigs were randomly divided into five groups, each consisting of 10 pigs. The groups were fed a basal diet with no supplementation (control) or a diet supplemented with 1 gGP/kg (E1), 5 gGP/kg (E2), 10 gGP/kg (E3), or 15 gGP/kg (E4). The doses were selected based on preliminary tests. The pigs were slaughtered after 90 days, and their duodenum, jejunum, ileum, and caecum were sampled. We recommend a dose of 10 gGP/kg because it leads to many beneficial effects, including a significantly increased villous height, intestinal crypt depth, and V/C ratio in the duodenum and jejunum (p < 0.05). It also decreased the expression of pro-inflammatory markers such as IL-1β in the ileum and caecum, TNF-α in multiple intestinal segments, and MHC-II in the duodenum, jejunum, and caecum. Additionally, antioxidant activity was promoted through the increased immunohistochemical expression of Nrf2 and reduced NF-kB p65 expression. Growth performance also improved, with significantly higher ADG (p = 0.01) and ADFI values (p < 0.01) than those in the control group. In conclusion, polyphenol-rich grape pomace can be used as a supplement in fattening pig diets to maintain their health and productive performance. Full article

This study aimed to evaluate the intrinsic in vitro performance of naïve porcine prepubertal Sertoli cells (SCs) and SCs loaded with blank poly(lactic acid) microparticles (MP) or amphotericin B poly(lactic acid) microparticles (AmB-MP) against Candida tropicalis, a prevalent pathogenic non-albicans species. The objective was to assess their impact on biofilm formation and the cellular response mechanisms involved, building on previous findings that highlight SCs’ potential as anti-infective agents and drug carriers. Our results demonstrated that SCs successfully internalized Candida tropicalis while maintaining viability and exhibited a strong anti-infective effect, inhibiting biofilm formation by 70%. This inhibition increased to 80–90% when SCs were combined with AmB-MP. The interaction between SCs (both naïve and MP-loaded) and Candida tropicalis triggered the activation of MAPK, AKT, and NF-kB signaling pathways, leading to the upregulated expression of innate immune factors such as MHC-II, TLR-4, TGF-β, IDO, and β-defensin 123. These findings reinforce the role of SCs in infection control and drug delivery. Furthermore, their anti-infective and scavenging activity is linked to a tolerogenic phenotype, suggesting a potential dual therapeutic role at the host–pathogen interface. Full article

Tilapia Parvovirus (TiPV) is a rising pathogen responsible for high mortality in tilapia aquaculture. Understanding TiPV’s pathogenesis is crucial for developing effective management strategies. This study aimed to elucidate TiPV pathogenesis by evaluating its cytotoxic effects on Danio rerio gill (DRG) cell monolayers and its impact on host immune responses. PCR-confirmed TiPV-infected DRG cell monolayers were subjected to an MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay at 24, 48, 72, and 96 h post-infection to assess cell viability and cytotoxicity. The MTT assay revealed a progressive decline in DRG cell viability over time, with viable cell percentages decreasing from 66.71% at 24 h to 31.28% at 96 h in TiPV-infected cultures, compared to consistently high viability in controls. Simultaneously, quantitative real-time PCR (qPCR) was used to assess the expression of key immune-related genes, including Interleukins (IL-1β, IL-8), Toll-like receptor 7 (TLR7), Major Histocompatibility Complex II (MHC-II), Tumor Necrosis Factor α (TNF-α), Nuclear Factor Kappa B (NF-κB), and Chemokine Receptors (CRs).qPCR analysis showed an upregulation of IL-8, IL-1β, TNF-α, and CRs, indicating an early inflammatory response. However, significant downregulation of TLR7, MHC-II, and NF-κB suggests TiPV’s ability to modulate host immune responses. The results highlight that TiPV induces significant cytotoxicity in DRG cells, leading to severe cellular damage. The virus also alters host immune responses by modulating the expression of key immune genes, which may contribute to its virulence and persistence. These findings enhance our understanding of TiPV pathogenesis and highlight the need for targeted research to develop effective control strategies for TiPV in aquaculture systems. Full article

The largemouth bass ranavirus (LMBRaV) caused significant mortality and economic loss in the largemouth bass aquaculture industry around the world, including China. Vaccination is an efficient method for virus defense. In this study, an inactivated LMBRaV vaccine was prepared, and the prevention effect as well as the immune responses were analyzed after the primary and the secondary immunization. Compared to the control group, the counts of leucocytes and erythrocytes increased and peaked at day 14 after the primary immunization, and the proportions of leucocytes, including lymphocytes, monocytes, and neutrophils, were also up-regulation after the primary immunization. Serum neutralizing antibody titers increased and peaked (1:128) at day 28 after the primary immunization. Following the secondary immunization, antibody titers were increased to a higher level (1:512) at 28 days after the secondary immunization. Quantitative real-time PCR analysis demonstrated varying degrees of up-regulation of mhc II, igM, il-1β, and cd8α transcriptions in the head kidney, which showed that innate and adaptive immune responses were both induced after the primary and the secondary immunization. After challenge with LMBRaV, the relative percent survival rates (RPS) for primary and secondary immunization with inactivated LMBRaV vaccine were determined to be 62.92% and 95.51%, respectively. Therefore, this study suggests that utilizing an inactivated LMBRaV vaccine could induce efficient immune responses and antibody, which might provide a potential efficient countermeasure for LMBRaV prevention. Full article