Search Results (30,206)

Background: Experimental autoimmune thyroiditis is an important animal model for studying Hashimoto’s thyroiditis. Our aim was to develop the model using CBA/J-DR3 mice expressing human HLA-DR3, which is associated with autoimmune thyroiditis in humans, to better simulate human autoimmune thyroiditis. Such a humanized model can be used to test specific antigen therapies for autoimmune thyroiditis. Methods: CBA/J-DR3 mice were produced by back-crossing B6-DR3 mice to the CBA/J background. Female CBA/J-DR3 mice were immunized with human thyroglobulin (Tg) in complete Freund’s adjuvant on days 0 and 7. On day 21, mice were sacrificed, blood collected, spleen and thyroid harvested for analysis. Splenocytes were analyzed for T cell responses to Tg and its major T-cell epitope in human autoimmune thyroiditis, Tg.2098. Serum anti-thyroglobulin antibodies were measured by ELISA, and thyroid-stimulating hormone was measured using the Luminex assay. Thyroid histology and immunohistochemistry were examined. Results: Immunized CBA/J-DR3 mice showed significant T cell proliferation in response to Tg (stimulation index 3.4 ± 4.5) and Tg.2098 (1.5 ± 0.7). Anti-thyroglobulin antibody levels were elevated in immunized mice when compared to control mice (2.05 ± 0.75 vs. 0.15 ± 0.06, p < 0.0001). T cells demonstrated higher reactivity to thyroid antigens by enhanced production of pro-inflammatory cytokines. Thyroid immunohistochemistry revealed mild CD3-positive T-cell infiltration. Conclusions: This novel humanized CBA/J-DR3 mouse model of Hashimoto’s thyroiditis demonstrates key features of human autoimmune thyroiditis. The HLA-DR3 background and the immune response to Tg and Tg.2098 enhance translational relevance, making this a valuable model for studying thyroid disease pathogenesis and testing targeted immune-modifying therapies. Full article

Introduction: Parkinson’s disease can influence oral health by impairing motor function and altering salivary composition, potentially affecting the oral microbiome. Materials and Methods: The objectives of this study are fourfold: (a) to compare the prevalence of bacterial species associated with periodontal disease in patients with and without Parkinson’s disease (PD), (b) to assess whether the coexistence of periodontal disease in PD patients contributes to an imbalance in the oral microbiome, (c) to evaluate the correlation between periodontal clinical indices (plaque index, tartar index, bleeding index, and probing depth) and the concentrations of specific periodontopathogenic bacterial species, and (d) to explore the potential implications of these evidences for clinical management and preventive strategies in Parkinson’s patients. The main objective of this study is to compare periodontal clinical indices (plaque index, tartar index, bleeding index, and probing depth) and the bacterial profile of patients with periodontal and Parknson’s disease. Two groups were included: 15 patients with periodontal disease (control group) and 16 patients with both periodontal and Parkinson’s disease (study group). Microbial samples were collected from the periodontal pockets at baseline and analyzed using the Polymerase Chain Reaction (PCR) Perio-Ident 12 kit to detect major periodontal pathogens. Results: Periodontal indices showed no statistically significant differences between groups, although the study group presented lower mean tartar index (49.31% vs. 67.4%, p = 0.069), bleeding on probing (44.31% vs. 56.67%, p = 0.137), and plaque index (66% vs. 68.93%, p = 0.754). Median bacterial loads were generally higher in control group, with Tannerella forsythia, but without statistically significant difference (p = 0.072). Significant correlations between plaque index and multiple pathogens occurred only in control gorup, suggesting disrupted plaque–pathogen dynamics (p < 0.05). Conclusions: The results highlight the potential value of integrating clinical and microbiological assessment when managing periodontal disease in patients with Parkinson’s disease. Full article

The polymorphism of the bovine β-casein gene (CSN2) is of increasing interest due to its relevance for A2 milk production. This study genotyped 2773 Holstein-Friesian cows for five CSN2 alleles (A1, A2, A3, B, I) using both conventional DNA-based PCR and a newly evaluated direct PCR protocol. Eleven genotypes were detected, with A2/A2 (33.9%) and A1/A2 (30.3%) being the most common, resulting in an A2 allele frequency of 59.0%. Genetic diversity indices indicated moderate polymorphism and a significant deviation from Hardy–Weinberg equilibrium, consistent with ongoing selection for the A2 allele. Associations between CSN2 genotype and milk traits (305-day milk, fat, and protein yield; fat% and protein%) were evaluated using linear mixed-effects models including lactation number, age at calving, and calving year as covariates, and cow ID as a random intercept. Several genotype effects reached statistical significance (p < 0.05); however, all effect sizes were very small (partial η² < 0.01), indicating that any influence of CSN2 on production traits is negligible within this population and management context. These findings suggest that A2-oriented selection is unlikely to compromise productivity. The direct PCR genotyping method achieved 96–100% success and enabled substantially faster and more cost-efficient processing (approximately 80–90% reduction in reagent costs), providing a rapid and scalable approach for large herds. Full article

Background/Objectives: Diffuse large B-cell lymphoma (DLBCL) is an aggressive and heterogeneous malignancy, for which predicting clinical outcomes remains challenging. Although immune-checkpoint pathways are known to influence tumor biology, the impact of their germline variants on DLBCL susceptibility and prognosis has not been fully elucidated. Methods: Variants in PD-L1 gene CD274 (rs4143815, rs822336), and miR-155 gene MIR155HG (rs767649, rs1893650), assessed by TaqMan assays in 99 DLBCL patients and 113 age- and sex-matched healthy controls, were associated with clinicopathological features, treatment response, overall survival (OS), relapse-free survival (RFS), and disease susceptibility. Results: The PD-L1 variant rs822336 was significantly associated with relapse status (p = 0.005) and RFS (p = 0.008), with the wild-type GG genotype showing the poorest RFS that remained independent in the multivariate Cox analysis (HR = 2.387, p = 0.003). Conversely, rs4143815 showed a nominal association with treatment resistance (p = 0.026), while patients carrying the GG genotype had worse OS (p = 0.006). In susceptibility analyses, miR-155 variant rs767649 showed a nominal association with DLBCL risk, with the rare AA genotype showing an increased risk of DLBCL (OR = 5.234, p = 0.045), which did not remain significant after Bonferroni correction. Conclusions: In a hypothesis-generating manner, these findings suggest that PD-L1 genetic variants may predominantly influence disease progression and outcomes, while miR-155 variation may contribute to DLBCL susceptibility. These findings highlight germline immunogenetic variants as stable, treatment-independent markers that may inform future studies on risk stratification and prognosis in DLBCL. Full article

Prolonged exposure to ultraviolet (UV) radiation in sunlight is a major extrinsic factor that impairs skin function and accelerates photoaging. In this study, a murine model of ultraviolet B (UVB)-induced photoaging exhibited characteristic symptoms, including skin roughness, erythema, hyperpigmentation, and increased wrinkle formation. Epicatechin gallate (ECG), a natural flavonoid, has demonstrated potential skin-protective properties. However, its specific effects and mechanisms against UVB-induced photoaging are not fully understood. Here, we investigated the protective role and underlying mechanism of ECG against UVB-induced damage in human epidermal keratinocytes (HaCaT cells). Using network pharmacology, p38 mitogen-activated protein kinase (p38 MAPK), specifically the p38α isoform, was identified as a key potential target of ECG. Our experimental results confirmed that ECG significantly attenuated UVB-induced photoaging. Mechanistically, ECG treatment effectively suppressed UVB-triggered phosphorylation of p38α, promoted autophagic flux (as evidenced by increased LC3B conversion and decreased p62 levels), and substantially reduced intracellular reactive oxygen species (ROS) accumulation. Consequently, ECG mitigated mitochondrial dysfunction, restored normal cell cycle progression, and decreased the expression of senescence-associated markers (p53, p16, p21) and inflammatory cytokines (IL6, TNF-α). In summary, our findings demonstrate that ECG protects against UVB-induced photoaging primarily by inhibiting p38α activation, thereby enhancing autophagy and alleviating oxidative stress. This study positions ECG as a promising therapeutic candidate for preventing and treating skin photoaging. Full article

The development of effective vaccines is a critical step in effective disease management in aquaculture. This study introduces a novel Multiepitope Chimeric Vaccine (MCV) designed to enhance immunity in lumpfish against Vibrio anguillarum, Aeromonas salmonicida, Yersinia ruckeri, Moritella viscosa and Piscirickettsia salmonis. Epitopes from major toxins and virulence factors were selected to construct the MCV in silico. Structural validation showed 96.7% of residues in favored regions, confirming stability. Codon optimization yielded a G+C content of 54.61% and a Codon Adaptation Index (CAI) of 1, indicating strong expression potential in Escherichia coli. Immune simulations predicted robust B- and T-cell responses, suggesting induction of both humoral and cell-mediated immunity. Experimental vaccination of lumpfish (n = 35/group) with E. coli-expressed MCV led to significantly elevated IgM levels at four- and six-weeks post-vaccination (p ≤ 0.05, p ≤ 0.01, respectively). Upon pathogen challenge, vaccinated groups showed delayed mortality against V. anguillarum, A. salmonicida, and P. salmonis, though survival differences were not statistically significant across treatments. These results highlight the immunogenicity potential of the MCV and its capacity to elicit targeted immune responses. However, further optimization is necessary to improve protective efficacy and survival outcomes. This study lays a foundation for the application of multiepitope vaccines in lumpfish aquaculture and supports ongoing efforts toward sustainable disease control strategies. Full article

Background: The human leukocyte antigen (HLA) is crucial for antigen presentation and vaccine efficacy. This study examined the association between HLA polymorphisms and the immune response to hepatitis B vaccination in children with acute lymphoblastic leukemia (ALL). Methods: 101 pediatric ALL patients at Shanghai Children's Medical Center affiliated with Shanghai Jiaotong University School of Medicine who tested negative for hepatitis B surface antibody (anti-HBs) and were not infected with hepatitis B received three doses of the hepatitis B vaccine. Anti-HBs titers were measured before and after vaccination. Participants were divided into high- and low-response groups based on post-vaccination anti-HBs titers. Sequence-specific primer polymerase chain reaction (PCR-SSP) was used to genotype HLA-A, -B, -Cw, -DRB1, and -DQB1 alleles. Results: Pre-vaccination anti-HBs titers were 3.38 ± 2.97 mIU/mL, and the post-vaccination seroconversion rate was 100% with mean titers of 429.61 ± 303.13 mIU/mL (p < 0.001). Following immunization, the low-response group (11.88%) had an anti-HBs titer of 56.47 ± 28.38 mIU/mL, while the high-response group (88.12%) had an anti-HBs titer of 479.93 ± 287.70 mIU/mL. There were significant differences in allele frequencies of B*3501 and Cw*0303 between the two response groups (p < 0.05). Binary logistic regression analysis showed that the B*3501 allele was negatively correlated with the anti-HBs response level (p < 0.05). Conclusions: HLA-B*3501 may be associated with lower antibody response levels in children with ALL who completed the full hepatitis B vaccination series. All these children demonstrated protection against the hepatitis B virus (HBV). We will subsequently validate the association between HLA-B*3501 and the level of hepatitis B vaccine immune response in children with ALL through expanding the sample size or conducting a multicenter study. Full article

Acute liver failure (ALF) is a rapidly progressive and life-threatening condition with limited pharmacological interventions. Poria cocos, a medicinal fungus widely used in traditional Chinese medicine, has been reported to exhibit anti-inflammatory and hepatoprotective activities; however, its potential involvement in ALF remains incompletely understood. In this study, an integrative exploratory strategy combining network pharmacology, molecular docking, and in vivo experiments was employed to investigate the protective effects of Poria cocos against lipopolysaccharide/D-galactosamine (LPS/D-GalN)-induced ALF in rats. Rats were pretreated with Poria cocos extract (50 or 200 mg/kg), and hepatoprotective effects were assessed by survival analysis, serum biochemical indicators(alanine aminotransferase [ALT], aspartate aminotransferase [AST], total bilirubin [TBil], and international normalized ratio [INR]), histopathology, and expression of inflammation- and PI3K/AKT-related markers. Network pharmacology analysis identified fifteen putative bioactive components of Poria cocos and 178 ALF-related overlapping targets, with enrichment analyses highlighting multiple inflammation-, apoptosis-, and PI3K/AKT-related signaling pathways. Molecular docking suggested potential interactions between major components and predicted core targets. In vivo, Poria cocos pretreatment was associated with improved survival, alleviated liver injury, and reduced the expression of inflammatory and apoptosis-associated markers, including PI3K, AKT1, NF-κB, TNF-α, MAPK14(p38), Caspase-3, and MMP2. Taken together, network pharmacology analysis identified PI3K/AKT-associated signaling as a candidate pathway, and the in vivo findings were generally consistent with this prediction, suggesting that the hepatoprotective effects of Poria cocos may involve multi-target regulation of inflammation- and apoptosis-related pathways. Full article

Bacillus atrophaeus has considerable potential for development as a microbial pesticide. Optimization of fermentation conditions and the wettable powder (WP) formulation is critical for its industrialization and application in sustainable agriculture. In this study, the fermentation of B. atrophaeus YL84 was optimized using single-factor experiments and response surface methodology. Based on these results, a WP formulation was developed and further optimized. The optimal carbon, nitrogen, and inorganic salt sources were sucrose (13.9 g·L⁻¹), tryptone (11.8 g·L⁻¹), and MgSO₄ (5.9 g·L⁻¹), respectively; optimal fermentation conditions were pH 7.0, 32 °C, and 210 r·min⁻¹. After optimization, the inhibition rate and OD₆₀₀ reached 83.71% and 1.758, respectively. The optimized formulation comprised attapulgite-based powder (79%, as carrier), sodium alkyl naphthalene sulfonate (5.4%) as a wetting agent, PEG-6000 (12.6%), CaCO₃ (2%), and vitamin C (1%). The resulting WP exhibited a spore viability of 2.63 × 10⁹ CFU·g⁻¹, and its 50-fold dilution demonstrated antagonistic activity in vitro against Cytospora pyri (Korla pear valsa canker agent) and biocontrol efficacy in vivo on detached-branch assays. These findings demonstrate that the YL84 WP is a promising candidate for the biological control of Korla pear valsa canker. Full article

Epithelial-mesenchymal transition (EMT) is a key driver of invasion, metastasis, and treatment resistance in gastric cancer, yet its post-transcriptional regulation by microRNAs (miRNAs) is not fully delineated. We performed a structured literature search in PubMed, Web of Science, and Scopus for studies evaluating miRNAs in relation to EMT in gastric cancer and synthesised tumor-intrinsic, microenvironmental, and circulating EMT-related miRNA networks. Downregulated, predominantly tumor-suppressive miRNAs, including miR-34a, miR-200 family, miR-148a, miR-204, miR-30a, miR-101, miR-218, miR-26a, miR-375, miR-506, and others, converge on EMT transcription factors and pathways such as ZEB1/2, Snail, TGF-β/SMAD, Wnt/β-catenin, c-Met, and PI3K/AKT, and their restoration reverses EMT phenotypes in preclinical models. Upregulated oncomiRs, such as miR-21, miR-17-5p, miR-106b-5p, miR-23a, miR-130a-3p, miR-196a-5p, miR-181a, miR-616-3p, miR-301a-3p, miR-150, miR-27a-3p and miR-192/215, target tumor suppressors and reinforce these pathways. Cancer-associated fibroblast, macrophage, neutrophil, and natural killer cell-derived miRNAs, together with systemic indices such as the neutrophil-to-lymphocyte ratio and mediators like FAM3C, add microenvironmental layers of EMT regulation. Several EMT-related miRNAs show consistent associations with invasion, metastasis, peritoneal dissemination, prognosis, and chemoresistance, and many are detectable in circulation. Overall, EMT-related miRNAs orchestrate gastric cancer cell plasticity and tumor-microenvironment crosstalk and represent promising biomarker and therapeutic candidates that warrant validation in prospective, subtype-stratified, and translational studies. Full article

Background/Objectives: Pancreatic ductal adenocarcinoma (PDAC) exhibits poor clinical response to gemcitabine, largely due to intrinsic and acquired mechanisms of chemoresistance. Identifying agents capable of enhancing gemcitabine efficacy without increasing cytotoxicity remains an unmet therapeutic need. Here, we characterise a small drug sensitiser molecule, B12, and evaluate its potential to sensitise PDAC cells to gemcitabine. Methods: Gemcitabine’s dose–response was assessed by MTT assay to determine IC₅₀ values and dose-modifying factor (DMF). Phenotypic consequences of co-treatment were examined using colony formation and wound scratch assays. Mitochondrial membrane potential (JC-1) and apoptosis (Annexin V/PI) were measured using flow cytometry. Transcriptomic profiling was performed using mRNA-seq with differential expression analysis and pathway enrichment (KEGG/GSEA). NF-κB activity was assessed by nuclear and cytoplasmic fractionation of p65, and RT-qPCR validation of NF-κB associated target genes. Results: B12 alone displayed minimal cytotoxicity in the PANC-1 cell line and normal pancreatic ductal HPDE cells, yet shifted the gemcitabine dose–response curve in PANC-1 cells, reducing the IC₅₀ and yielding a dose-modifying factor of 1.39. Functionally, B12 enhanced gemcitabine-induced suppression of colony formation and reduced wound closure relative to gemcitabine alone. The co-treatment also increased both mitochondrial depolarisation and apoptotic cell populations, with increased cell proliferation inhibition over time. Transcriptomic profiling identified a set of B12-associated genes downregulated both in B12-treated and B12 + gemcitabine conditions, including factors linked to growth, survival, inflammation, metabolism, and drug inactivation. Gene set enrichment analysis revealed negative enrichment of NF-κB associated pathways during B12 co-treatment. Consistently, nuclear-cytoplasmic fractionation showed that B12 reduced gemcitabine-induced nuclear accumulation of p65, accompanied by decreased expression of NF-κB associated targets such as BCL2L1, CCL20, SLC2A1, and MAP3K14. Conclusions: In PDAC cell models, B12 enhances gemcitabine cytotoxic response while displaying minimal intrinsic toxicity under the conditions tested. The sensitising phenotype is accompanied by increased apoptotic susceptibility and is associated with reduced NF-κB signalling at the pathway, transcript, and p65 nuclear localisation levels. However, to establish causality, the lack of sensitisation in HPDE cells will require further validation. Full article

Background/Objectives: Accurate risk stratification at diagnosis is crucial for the optimal management of diffuse large B-cell lymphoma (DLBCL). While Fluorine-18 fluorodeoxyglucose positron emission tomography/computed tomography ([¹⁸F]FDG PET/CT) is the standard imaging modality for staging, the prognostic value of non-tumoral uptake patterns remains under investigation. Here, we aimed to investigate the prognostic significance of qualitative splenic [¹⁸F]FDG uptake on baseline PET/CT in patients with newly diagnosed DLBCL, focusing on relapse-free survival (RFS) and overall survival (OS). Methods: This retrospective study included consecutive patients with newly diagnosed DLBCL in a Korean cohort who underwent baseline [¹⁸F]FDG PET/CT between December 2016 and August 2023. Qualitative splenic uptake was visually assessed on maximum intensity projection images. Associations between splenic uptake, prognostic indices, and clinicopathologic characteristics were evaluated. Survival outcomes and independent prognostic factors were analyzed using Kaplan–Meier methods and Cox proportional hazards regression models. Results: A total of 142 patients were analyzed (43 relapsed, 58 died). Positive splenic [¹⁸F]FDG uptake was observed in 72 patients and was significantly more frequent in patients who relapsed (p < 0.001). Positive splenic uptake was significantly associated with inferior RFS (p < 0.001) and OS (p = 0.010). For RFS, advanced Ann Arbor stage, ECOG performance status, and extranodal involvement were also significant factors. In multivariable analysis, positive splenic uptake remained an independent predictor of poorer RFS (hazard ratio 2.175, p = 0.043), along with advanced stage (hazard ratio 2.872, p = 0.004). Conclusions: Qualitative splenic [¹⁸F]FDG uptake on baseline PET/CT is associated with adverse clinical outcomes in patients with DLBCL and serves as an independent prognostic factor for RFS. Full article

Lipopolysaccharides (LPSs) are potent pro-inflammatory neurotoxins abundant in the gut microbiome and originate primarily from Gram-negative bacteria, such as Escherichia coli. LPS levels increase with brain aging and accumulate around neurons in Alzheimer’s disease (AD) brains. Microbiome-generated LPS and other endotoxins cross gut barriers, enter systemic circulation, and translocate across the blood–brain barrier into vascularized brain regions. These processes are exacerbated by aging and neurovascular diseases. Although pro-homeostatic systems mitigate LPS effects, these defenses can fail. This study provides the first evidence that acyloxyacyl hydrolase (AOAH; EC 3.1.1.77), a microglia-enriched LPS detoxifying enzyme, shows reduced expression in AD brain tissue. Analysis of AD patient brains revealed reduced AOAH messenger RNA (mRNA) levels, accompanied by elevated expression of microRNA (hsa-miR-450b-5p), an inflammation regulator. Furthermore, luciferase reporter assays demonstrated that miR-450b-5p specifically targets the AOAH 3′-UTR, leading to a dose-dependent suppression of reporter activity. Also, in vitro experiments on human neuronal glial (HNG) cells further confirmed down-regulation of AOAH expression at protein levels by miR-450b-5p. These findings suggest miR-450b-5p-mediated AOAH deficiency drives LPS-associated neurotoxicity and inflammatory neurodegeneration in AD. Full article

Outer membrane vesicles (OMVs) are tractable, cell-free microbial outputs that can shape innate immune programs. Here, we compared OMVs from the probiotic Escherichia coli Nissle 1917 (EcN) and the commensal strain ECOR12 in a paired within-donor model of human monocyte-derived dendritic cells (Mo-DCs) (N = 20). In the core integrated arm, Mo-DCs were exposed to iDC control, EcN OMVs, or ECOR12 OMVs (10 µg/mL, 24 h) and profiled for maturation markers (CD14, CD83, CD209), cytokines (IL-6, TNF-α, IL-10), and a targeted miRNA panel (miR-155-5p, let-7i-3p, miR-146b-5p, miR-29a-5p). Both OMV types promoted maturation (increased CD83 and reduced CD14), but generated distinct cytokine–miRNA configurations, with ECOR12 tending toward an IL-10–high profile and EcN toward higher IL-6/TNF-α tendencies. Multivariate integration separated conditions into reproducible, strain-specific immune fingerprints, supporting the key take-home that probiotic versus commensal E. coli OMVs imprint distinguishable coordinated response states in human DCs. In an extended phenotyping arm, ECOR63 OMVs were evaluated by ELISA and flow cytometry only and were not included in miRNA profiling or integrated PCA due to unavailable matched miRNA measurements. Full article

Background/Objectives: SLC13A5 encodes a sodium–citrate cotransporter implicated in early-onset epileptic encephalopathy and metabolic brain dysfunction, yet its developmental regulation and molecular context in the human brain remain incompletely defined. Methods: Leveraging human developmental transcriptomes from the Evo-Devo resource, we delineated tissue trajectories and network context for SLC13A5 across the fetal–postnatal life. Results: In the cerebrum, SLC13A5 expression rises from late fetal stages to peak in the first postnatal year and then declines into adulthood, while cerebellar levels increase across the lifespan; liver shows a fetal decrease followed by sustained postnatal upregulation. A transcriptome-wide scan identified extensive positive and negative associations with SLC13A5, and a signed weighted gene co-expression network analysis (WGCNA) built on biweight midcorrelation placed SLC13A5 in a large module. The module eigengene tracked brain maturation (Spearman rho = 0.802, p = 8.62 × 10⁻⁶) and closely matched SLC13A5 abundance (rho = 0.884, p = 2.73 × 10⁻⁶), with a significant partial association after adjusting for developmental rank (rho = 0.672, p = 6.17 × 10⁻⁴). Functional enrichment converged on oxidative phosphorylation and mitochondria. A force-directed subnetwork of the top intramodular members (|bicor| > 0.6) positioned SLC13A5 adjacent to a densely connected nucleus including CYP46A1, ITM2B, NRGN, GABRD, FBXO2, CHCHD10, CYSTM1, and MFSD4A. Conclusions: Together, these results define a developmentally tuned, mitochondria-centered program that co-varies with SLC13A5 in the human brain across the lifespan. It may provide insights to interrogate age-dependent phenotypes and therapeutic avenues for disorders involving citrate metabolism. Full article