Search Results (4,621)

Sertoli cells (SCs) are essential for spermatogenesis and provide structural and nutritional support to germ cells in the Chinese Holstein cattle testis. Although microRNAs (miRNAs) are known to regulate SC function, the specific role of Bta-miR-146a in bovine SCs is unclear. This study investigated the mechanisms by which Bta-miR-146a regulates bovine immature SCs. Using molecular cloning, we constructed Bta-miR-146a overexpression and interference vectors and transfected them into SCs via lipofection. Quantitative real-time PCR (RT-qPCR), 5-ethynyl-2′-deoxyuridine (EdU) proliferation assays, Cell Counting Kit-8 (CCK-8) viability assays, and flow cytometry revealed that Bta-miR-146a overexpression inhibited SC proliferation and promoted apoptosis, whereas Bta-miR-146a inhibition increased proliferation and suppressed apoptosis. Dual-luciferase reporter assays confirmed that SMAD4 is a direct target of Bta-miR-146a; SMAD4 interference reduced SC proliferation and increased apoptosis, whereas overexpression had the opposite effect. Furthermore, activity of this gene modulates the TGFβ/MAPK signaling pathway; SMAD4 interference reduces the expression of TGFβ, TGF-βRII, DAXX, MAP3K5, P38, and MAX. These findings indicate that the Bta-miR-146a/SMAD4/TGFβ/MAPK axis is a key regulator of SC proliferation and apoptosis, offering insights into the molecular mechanisms underlying bovine spermatogenesis and potential targets for improving reproductive performance. Full article

Background/Objectives: Carotid intima–media thickness (CIMT) is widely recognized as an established biomarker of systemic atherosclerosis and coronary artery disease (CAD). Nevertheless, the connection between circulating miR-29 family members, CIMT and CAD severity has not yet been determined. The aim of this study was to investigate the association of plasma miR-29a and miR-29c with CIMT and CAD severity. Methods: A total of 628 patients scheduled for elective coronary angiography were included in the study. CAD severity was estimated using the Gensini scoring system and was classified into mild, moderate, and severe categories. Additionally, CIMT was evaluated using high-resolution B-mode ultrasonography, while plasma miR-29a and miR-29c levels were detected using qRT-PCR. Results: CIMT increased significantly with CAD severity (0.76 ± 0.15, 0.94 ± 0.18, and 1.12 ± 0.22 mm for mild, moderate, and severe CAD, respectively; p < 0.001). MiR-29a expression progressively increased, whereas miR-29c expression decreased with advancing CAD (both p < 0.001). MiR-29a correlated positively with CIMT (r = 0.612) and Gensini score (r = 0.658), while miR-29c showed negative correlations (r = −0.527 and −0.584, respectively; all p < 0.001). CIMT, miR-29a, and miR-29c were found to be independently associated with CAD severity. Their combined model demonstrated a powerful association with severe CAD (AUC = 0.927; sensitivity 88.6%; specificity 84.1%). Conclusions: Plasma miR-29a and miR-29c are independent biomarkers of CIMT and CAD severity. Their combination with CIMT can improve diagnosis of severe CAD and may enhance cardiovascular risk stratification. Full article

The purpose of the present study was to investigate how periodontitis and diabetes mellitus (DM) are associated with Alzheimer’s disease (AD) through microRNA (miRNA) using AD model mice. The experimental period was 8 weeks. Twenty-four male knock-in mice (B6-AppNL-G-F/NL-G-F/J) were divided into four groups: control group fed a normal diet (C), DM group fed a high-fat/sucrose diet (DM), periodontitis (P) group, and DM + periodontitis (DM+P) group. Memory performance was compared using the Y-maze test. Next-generation sequencing was performed on brain samples, and fold changes in miRNA expression were calculated by comparing the DM+P and C groups. Integrated miRNA–mRNA analysis identified putative miRNA-targeted mRNAs, and protein expression of the top candidate gene was assessed. Memory function in the DM+P group was significantly lower than in the C group. Among the seven mRNAs identified by the integrated analysis, Neurod1 showed the greatest decrease in expression, and it was predicted to be regulated by miR-693-3p. Neurod1 protein expression in the hippocampus was significantly lower in the DM+P group than the C group. Our results suggest that the combined exposure to periodontitis and DM was associated with AD-like pathological changes and identified the miR-693-3p/Neurod1 pair as a candidate regulatory axis. Full article

The earliest gut and nasal bacterial community profiles after birth remain incompletely characterized, particularly when both sites are examined in parallel. We aimed to characterize these profiles according to delivery mode within the first 48 h of life in a Mexican cohort. Meconium and nasal wash samples from 60 neonates (30 vaginal and 30 cesarean deliveries) were analyzed by V3–V4 16S rRNA gene amplicon sequencing on an Illumina MiSeq platform, followed by QIIME 2 processing. All 60 meconium samples yielded analyzable 16S rRNA profiles. Nasal washes yielded analyzable profiles in 10/30 vaginally delivered and 12/30 cesarean-delivered neonates (p = 0.79). No statistically significant differences in alpha diversity were detected by delivery mode in gut samples (observed ASVs, p = 0.842; Shannon index, p = 0.183; Pielou’s evenness, p = 0.121; Faith’s phylogenetic diversity, p = 0.574) or nasal samples (p = 0.356, 0.947, 0.644, and 0.114, respectively). In unadjusted PERMANOVA comparisons, gut community structure differed by delivery mode for Bray–Curtis (p = 0.001), weighted UniFrac (p = 0.021), and unweighted UniFrac (p = 0.013) distances, whereas nasal comparisons were not statistically significant. Traditional ANCOM identified Staphylococcus as the feature with the highest W statistic in cesarean-delivered neonates (W = 65). Descriptive phylum-level profiles were examined across maternal urinary tract infection, premature rupture of membranes, maternal comorbidities, maternal BMI, hours since birth, and recorded antibiotic exposure; these comparisons were not multivariable and cannot exclude residual confounding. Delivery mode was associated with differences in early gut bacterial community structure in this cohort. Nasal findings were inconclusive because only 22 washes yielded analyzable 16S rRNA profiles and the workflow was not optimized to quantify ultra-low-biomass bacterial DNA. These cross-sectional data do not establish persistence, causality, or the timing at which nasal communities become stable. Full article

Distinguishing relapsing–remitting multiple sclerosis (RRMS) from secondary-progressive disease (SPMS) remains a clinical challenge, as no single laboratory test reliably identifies the transition. We explored whether plasma extracellular-vesicle (EV) microRNAs could separate the two phenotypes. Four candidate EV-miRNAs: miR-30a-5p, miR-223-5p, miR-155-5p, and miR-146a-5p, were quantified by qRT-PCR in plasma-derived EVs from 13 healthy controls (HC), 7 RRMS, and 16 SPMS patients, normalized to miR-16-5p. EVs were characterized by electron microscopy, dynamic light scattering, and zeta potential, confirming vesicles of expected morphology, size, and negative surface charge. Two miRNAs were informative. miR-155-5p was significantly downregulated in both RRMS and SPMS versus controls but did not differentiate the phenotypes (AUC 0.52), behaving as a disease-general marker. In contrast, miR-223-5p was selectively reduced in SPMS, to approximately one-quarter of control abundance, and provided the only signal distinguishing progressive from relapsing disease (AUC 0.73; rank-biserial −0.46). miR-30a-5p and miR-146a-5p were uninformative. These findings, consistent with the circulating/EV literature, suggest EV-miR-223-5p as a candidate marker of the progressive phenotype and EV-miR-155-5p as a disease indicator. Given the small cohort and near-detection-limit measurements, the study is hypothesis-generating and requires validation in larger, longitudinal cohorts. Full article

The Northeast black soil region is a major grain-producing area in China, where county-scale assessment of topsoil quality is essential for soil conservation and provides a potential indicator framework for regional soil quality monitoring. In this study, 500 composite cultivated-layer samples were collected from dryland croplands in Keshan County, Heilongjiang Province. Soil physical properties, basic chemical properties, and macro-, secondary, and micronutrient contents were measured to establish a total data set (TDS). Three hierarchical total data sets (TDS1, TDS2, and TDS3) were established from the original TDS according to the progressive incorporation of soil physical properties, basic chemical properties, macronutrients, secondary nutrients, and micronutrients. Within each hierarchical TDS, key indicators were selected using random forest (RF), mutual information (MI), principal component analysis (PCA), and minimum spanning tree (Tree) methods to construct algorithm-specific minimum data sets (MDSs). TDS1 included soil physical properties, basic chemical properties, and macronutrients; TDS2 further incorporated secondary nutrients; and TDS3 additionally included micronutrients to represent progressively comprehensive soil nutrient information. The soils exhibited considerable soil organic matter and cation exchange capacity, with mean values of 51.45 g kg⁻¹ and 34.85 cmolc kg⁻¹, respectively, and a mean pH of 6.04. Across the four algorithms, commonly retained indicators included SOM, TN, pH, CEC, and Silt, while nutrient-related indicators such as AP, AK, S, Zn, and Fe were additionally selected under different hierarchical MDSs, reflecting the importance of multi-nutrient information in soil quality characterization. Available phosphorus, available potassium, sulfur, and zinc showed greater spatial variability than basic physicochemical properties. The four algorithms differed in indicator selection, reflecting their distinct sensitivities to linear variation, information gain, multilevel contributions, and network structure. RF_MDS1 showed the highest agreement with the TDS (R² = 0.924), indicating its strong capability in preserving overall soil quality information using a simplified indicator set. RF_MDS3 maintained a high consistency with the TDS (R² = 0.836) while incorporating additional secondary nutrients and micronutrients. Therefore, RF_MDS3 was considered a more comprehensive MDS framework when multi-nutrient representation and potential nutrient constraint identification were prioritized, whereas RF_MDS1 remained an efficient option for simplified soil quality assessment. This framework may provide a transferable approach for soil quality assessment and nutrient management in comparable black-soil regions and dryland farming systems. Full article

Pleurotus ostreatus is an important edible mushroom, and its growth and development are affected by genetic and epigenetic mechanisms. miRNAs play an important role in fungal epigenetic mechanisms. In previous studies, Po-3KAT was identified as a candidate target of Po-milR-1, and its product was a substrate for ergosterol synthesis. However, the potential relationship between Po-milR-1 and Po-3KAT has not been studied in P. ostreatus. Therefore, this study aims to explore the potential role of Po-milR-1 and Po-3KAT in P. ostreatus. By integrating the Po-milR-1 interference vector into the genome of P. ostreatus, it was found that the mycelial growth rate was accelerated and the fruiting body phenotype changed in the Po-STTM transformants. The expression analysis during development suggested an inverse expression pattern between Po-milR-1 and Po-3KAT. The analysis of qRT-PCR and HPLC showed that 2 mmol/L H₂O₂ promoted the expression of Po-3KAT, and the content of ergosterol increased to 1.17-fold, while 5 mmol/L H₂O₂ inhibited the expression of Po-3KAT, and the content of ergosterol decreased to 0.82-fold. In contrast to oxidative stress, 100 mmol/L and 250 mmol/L salt stress inhibited the expression of Po-3KAT, and the content of ergosterol decreased to 0.87-fold and 0.94-fold, respectively. These results suggest that Po-milR-1 and Po-3KAT may be involved in the regulation of growth, development, and stress responses in P. ostreatus, and may be associated with the change in ergosterol content. Full article

A coordinated extracellular matrix (ECM) and cytoskeletal remodeling are critical drivers of hepatocellular carcinoma (HCC) progression, yet the upstream post-transcriptional mechanisms regulating these processes in spontaneous canine HCC remain poorly defined. Previously, we reported significantly downregulated miRNAs and enriched pathways from differentially expressed genes (DEGs) in canine HCC tissues. In the present study, we investigated putative post-transcriptional target interactions between the prioritized miRNAs and enriched ECM–cytoskeleton pathway-associated genes (n = 34). Integrative bioinformatic analysis prioritized ACTG1 and SDC1 as the key putative targets of cfa-miR-10a based on their concordant prediction by four target-prediction algorithms, high transcript abundance, and inverse correlations with cfa-miR-10a-5p. RNAhybrid and miRanda analyses supported favorable predicted interactions between cfa-miR-10a and the 3′UTRs of both transcripts, with RNAhybrid minimum free energy values ≤ −20 kcal/mol and miRanda scores ≥ 140. In the matched canine transcriptomic subset, cfa-miR-10a and its predicted targets showed inverse expression trends. Cross-species analysis using TCGA-LIHC datasets further demonstrated concordant dysregulation of ACTG1, SDC1 and hsa-miR-10a-5p in human HCC, despite their weak or non-concordant correlation with hsa-miR-10a-5p. Collectively, these exploratory findings identify a candidate cfa-miR-10a–ACTG1/SDC1 network associated with coordinated ECM and cytoskeletal transcriptomic alterations in canine HCC. While cross-species analysis supports the conservation of target gene dysregulation, the divergent miRNA-mRNA correlation patterns highlight potential species-specific post-transcriptional co-expression landscapes that warrant future functional validation. Full article

Background/Objectives: While currently termed premature ovarian insufficiency (POI), premature ovarian failure (POF) remains a prominent driver of female infertility with a molecular pathogenesis that is still lacking comprehensive clarification. In in vitro studies, the pathology of POI is commonly simulated through a well-characterized model involving VCD (4-vinylcyclohexene diepoxide)-mediated cytotoxicity within KGN-derived human granulosa cells. However, the key regulatory molecular networks involved in this process are still poorly characterized. Although microRNAs (miRNAs) have emerged as critical regulators in ovarian function decline, the specific role and underlying mechanism of miR-27b-3p in POI remain elusive. Methods: A VCD-induced KGN cell injury model was established by treating cells with 1.0 mM VCD for 24 h. Cell viability, apoptosis rate, and miR-27b-3p expression were assessed by CCK-8 assay, flow cytometry, and RT-qPCR, respectively. Overexpression and targeted suppression of miR-27b-3p were achieved by introducing its specific mimics and inhibitors, respectively. Target identification was conducted via bioinformatic prediction, EdU incorporation, Western blot, and dual-luciferase reporter assays. Functional rescue experiments were carried out by co-transfection with a PAPPA-overexpressing plasmid (oe-PAPPA). IGF-1 secretion was quantified by ELISA, and phosphorylation of IGF1R and AKT was analyzed by Western blot to determine whether miR-27b-3p modulates cellular phenotypes via the PAPPA–IGF-1–PI3K/AKT axis. Exogenous IGF-1 supplementation was further applied to confirm pathway dependence. Results: VCD treatment dose-dependently restrained cellular growth and stimulated apoptotic pathways in KGN cells; paralleling these phenotypic changes, miR-27b-3p abundance was remarkably increased. Ectopic expression of miR-27b-3p exacerbated VCD-induced growth inhibition and apoptosis, whereas its inhibition conferred cytoprotective effects. Through the integration of computational predictions and dual-luciferase reporter systems, PAPPA was definitively established as a direct downstream target of miR-27b-3p. miR-27b-3p negatively regulated both PAPPA mRNA and protein levels, thereby impairing PAPPA-mediated cleavage of IGF-binding proteins (e.g., IGFBP4) and subsequent release of free IGF-1. This led to reduced IGF-1 secretion and significantly diminished phosphorylation of IGF1R and AKT. Remarkably, PAPPA overexpression effectively reversed the detrimental effects of miR-27b-3p, and exogenous IGF-1 supplementation similarly attenuated miR-27b-3p–mediated proliferation arrest and pro-apoptotic phenotypes. Conclusions: This study uncovers a novel pathogenic mechanism whereby miR-27b-3p exacerbates VCD-induced granulosa cell injury by directly targeting PAPPA, suppressing IGF-1 release, and consequently inhibiting the PI3K/AKT pro-survival signaling pathway. A novel perspective on the fundamental basis of POI is established by this research, which further posits therapeutic manipulation of the miR-27b-3p/PAPPA/IGF-1 module as a prospective treatment for disrupted ovarian function. Full article

MicroRNAs (miRNAs) are key regulators of gene expression that act primarily by binding to target messenger RNAs (mRNAs). However, the biology of miRNAs is more complex than initially thought, with functional complexity extending beyond canonical sequences. A multilayered miRNA regulatory landscape involving isomiR generation, altered 5p/3p strand usage, arm switching, A-to-I RNA editing, and epitranscriptomic RNA modifications operates in eukaryotic cells to regulate miRNA function. Collectively, these mechanisms expand the functional diversity of miRNAs by regulating their biogenesis, stability, strand selection, and target specificity, increasing their functional plasticity and contributing to regulatory heterogeneity found in cells. IsomiRs arise from alternative Drosha/Dicer processing, terminal nucleotide additions, RNA editing, and genetic variation, producing functionally distinct isoforms. Arm switching alters gene regulatory outputs through context-dependent changes in predominant 5p/3p strand usage. In addition, epitranscriptomic RNA modifications, such as m6A and m5C, together with A-to-I RNA editing, represent an additional layer of miRNA regulation. These mechanisms can act directly on miRNAs or their precursors, or indirectly by modifying circRNAs and lncRNAs, thereby altering miRNA availability and function. Together, these processes form a dynamic regulatory network that influences key cancer hallmarks, including cell proliferation, apoptosis, epithelial–mesenchymal transition, metastasis, immune evasion, and therapy resistance. However, the contribution of these non-canonical regulatory layers to tumor-specific miRNA function remains poorly understood. In this review, we explore how isomiR generation, miRNA strand selection, arm switching, and epitranscriptomic regulation expand the functional diversity of miRNAs in gynecologic cancers. Full article

MicroRNAs (miRNAs) have emerged as promising biomarkers for treatment response prediction in breast cancer. However, the role of circulating and tumour-derived miRNAs in predicting pathological complete response (pCR) following neoadjuvant therapy in HER2-positive (HER2+) breast cancer remains incompletely defined. To systematically evaluate the association between miRNA expression profiles and pCR in HER2+ breast cancer undergoing neoadjuvant systemic therapy, including chemotherapy with HER2-targeted agents. A systematic review of PUBMED, EMBASE, SCOPUS, and WEB OF SCIENCE databases was performed according to PRISMA guidelines. Studies evaluating circulating or tumour-derived miRNAs associated with pCR following neoadjuvant therapy in HER2+ breast cancer were included. Methodological quality was assessed using the QUIPS tool. Nine studies involving 937 HER2+ breast cancer patients were included. Overall, 39 unique miRNAs and four circulating miRNA signatures were associated with pCR outcomes. Seven studies evaluated circulating miRNAs, while two assessed tumour-derived miRNAs. The increased expression of six circulating miRNAs, four circulating signatures, and seventeen tumour-derived miRNAs was associated with pCR. Conversely, the increased expression of eleven circulating miRNAs and five tumour-derived miRNAs was associated with residual disease and non-pCR. MiR-210 was the only miRNA associated with response across more than one study, with an increased expression associated with residual disease in both circulating and tumour-derived analyses. Several circulating and tumour-derived miRNAs demonstrate potential as predictive biomarkers of pCR following HER2-targeted neoadjuvant therapy. However, substantial heterogeneity and limited validation currently restrict clinical implementation. Future prospective multicentre studies using standardised methodologies are required to clarify the role of miRNA profiling in personalised treatment strategies for HER2+ breast cancer. Full article

This study aimed to examine position-specific associations of contextual factors (match outcome, match location, and opponent quality) and match running performance (MRP). The dataset comprised 1806 individual observations obtained from 160 matches with 509 players from 36 participating teams across the UEFA Champions League 2024/25 season. Metrics included total distance covered (TD), low-(LIR), moderate-(MIR), and high-intensity running (HIR), high-speed running (HSR), sprinting (SPR), and high-intensity accelerations (HIAs) and decelerations (HIDs). Significant playing position × match outcome interactions were observed for TD (F = 2.75, p = 0.005), MIR (F = 2.39, p = 0.015), HIR (F = 5.24, p < 0.001), HSR (F = 3.90, p < 0.001), and SPR (F = 3.69, p < 0.001). Significant playing position × opponent quality interactions were also identified for TD (F = 8.77, p < 0.001), LIR (F = 3.56, p = 0.007), MIR (F = 6.20, p < 0.001), HSR (F = 3.09, p = 0.015), and HIDs (F = 5.14, p < 0.001). In contrast, no significant playing position × match location interactions were observed. Match outcome, match location, and opponent quality explained 0.7–2.5% of overall variability in MRP (Part R² = 0.007–0.025). Therefore, these contextual factors should be considered as supplementary information that may improve the interpretation of position-specific MRP rather than as factors that substantially determine the physical demands imposed during match-play. Full article

Butyl benzyl phthalate (BBP), a widely used endocrine-disrupting chemical, has been associated with reproductive toxicity; however, its long-term effects during aging remain poorly understood. In the present study, we examined the effects of prolonged BBP exposure on the male reproductive system using naturally aged C57BL/6J mice. Mice received BBP at 169 μg/kg/day in drinking water for 10 or 22 months and were analyzed at 24 months of age. No significant differences in body weight, food intake, or water consumption were observed among the experimental groups. Representative gross images showed apparent distension and dark-red discoloration of the seminal vesicles in BBP-exposed aged mice. Compared with Young mice, the BBP-exposed aged groups showed higher expression of IL-1β, IL-6, TNFα, SOD1, and SOD2 and lower CAT expression in the seminal vesicle. H₂DCFDA fluorescence showed a non-significant increasing trend. Because an untreated age-matched Old group was not included in the seminal vesicle analyses, the effects of aging and BBP could not be distinguished. In contrast, the testis showed limited changes in inflammatory cytokine-, antioxidant enzyme-, and steroidogenesis-related gene expression and in H₂DCFDA fluorescence relative to the untreated Old group. Exploratory miRNA sequencing of pooled testicular RNA and comparison with a TM3 Leydig cell dataset identified miR-664-5p as a candidate showing higher relative abundance in both datasets. TargetScan analysis predicted binding sites for miR-664-5p in FOXO1 and FOXO3. In BBP-treated TM3 cells, FOXO3 protein expression was decreased, and FOXO6 expression was increased, whereas FOXO1 showed no consistent dose-dependent change. These molecular findings do not establish direct regulation of FOXO proteins by miR-664-5p or explain the seminal vesicle findings. Full article

Insects periodically undergo molting to replace their old epidermis for growth and morphological adaptation. Muscleblind-like (MBNL) is a multifunctional protein involved in miRNA processing. To explore the role of miRNA processing in insect molting, we characterized BmMBNL in Bombyx mori. BmMBNL exhibited spatiotemporal expression across tissues and developmental stages, with the highest levels in the epidermis and upregulation during molting. CRISPR/Cas9-mediated knockout of BmMBNL (BmMBNL-KO) caused molting defects at the 3rd and 4th instar larvae, resulting in reduced size and eventual lethality. Moreover, BmMBNL-KO individuals showed a thinner chitin layer between the new and old epidermises. RT-PCR and transcriptome analyses revealed upregulation of miR-1, miR-71, miR-263, miR-8-5p, and miR-2a-3p, which suppressed chitinase expression, ultimately leading to molting failure. This study provides novel insights into the potential correlation between BmMBNL, miRNA abundance and insect molting in Bombyx mori. Full article

Background: Frontotemporal dementia (FTD) is a neurodegenerative disease that shares numerous clinical features with other forms of dementia. In this context, non-coding RNAs, specifically microRNAs (miRNAs), represent a promising tool for differential diagnosis. Since these miRNAs can be isolated from circulating extracellular vesicles (EVs) in peripheral blood, they provide a direct insight into FTD-specific molecular processes. Consequently, while EV-contained miRNAs hold potential as disease-specific biomarkers, investigating their relative target genes can help elucidate their precise functional roles. Aim: This work aimed to identify a specific miRNA signature to better characterize FTD pathology. Methods: Building on a previous Next-Generation Sequencing (NGS) analysis, three candidate miRNAs were selected for validation in both EVs and peripheral blood mononuclear cells (PBMCs) of FTD patients. Subsequently, the predicted target genes of two of these miRNAs were validated in PBMCs to assess their expression levels. Results: Our findings revealed that miR-365a-3p and miR-212 were significantly down-regulated in FTD. Conclusions: Together with their target genes, these miRNAs are involved in cell cycle and apoptotic pathways, suggesting a potential role in the pathological mechanisms of the disease. Full article