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This study identified miR-5701 and TP53 through bioinformatics analysis utilizing the GEO, GeneCards, and miRDB databases. The serum expression levels were quantified via Reverse Transcription Quantitative Real-time Polymerase Chain Reaction (RT-qPCR). The binding relationship between miR-5701 and TP53 was confirmed using dual luciferase reporter assays. Subsequently, MC3T3-E1 cell differentiation was induced with Bone Morphogenetic Protein 2 (BMP2), and the expression changes of miR-5701 and TP53 after cell differentiation were detected. Subsequently, after overexpression of miR-5701 and TP53, the expression levels of key osteogenic differentiation markers Alkaline Phosphatase (ALP), Osteocalcin (OCN), and type I collagen were evaluated by ELISA. Serum miR-5701 expression is downregulated in osteoporotic fracture patients, while it increases after osteoblast differentiation. TP53 is a negative regulatory binding factor downstream of miR-5701. The mRNA expression of TP53 in the patient serum and the differentiated osteoblast cells is exactly opposite to that of miR-5701. Overexpression of miR-5701-mediated promotion of osteoblast proliferation and differentiation can be reversed by TP53. miR-5701 was found to facilitate osteoblast proliferation and differentiation through negative regulation of the target gene TP53 expression, suggesting that miR-5701 could serve as a promising therapeutic target for promoting fracture healing.

The present investigation was designed to assess the prognostic value of microRNA-648 (miR-648) in osteosarcoma (OS) and elucidate its regulatory mechanisms. Quantitative real-time PCR was employed to measure miR-648 expression levels in 80 paired OS specimens and their matched adjacent non-tumor tissues. Statistical assessments of clinical parameters were conducted using Chi-squared tests, while patient survival data were evaluated through Kaplan–Meier estimation and Cox proportional hazards regression modeling. Functional assays were performed in OS cell lines. Bioinformatic prediction of target genes was followed by experimental validation using dual-luciferase reporter assays. MiR-648 exhibited significant downregulation in OS clinical specimens and cell lines (p < 0.001). Low miR-648 expression correlated with lung metastasis (p = 0.027), advanced Enneking stage (p = 0.031), and poorer progression-free survival (p < 0.001). MiR-648 was identified as a significant independent prognostic indicator (hazard ratio [HR] = 0.235, p < 0.001). Moreover, the overexpression of miR-648 significantly suppressed cellular proliferation, migration capacity, and invasion potential while enhancing apoptotic activity (p < 0.001). High mobility group box 1 (HMGB1) was confirmed as a direct target, with its role in reversing miR-648's tumor-suppressive effects. MiR-648 exerts tumor-suppressive effects in OS by modulating HMGB1, suggesting its clinical utility as both a prognostic biomarker and a therapeutic intervention point.

Circulating microRNAs (miRNAs) are promising prostate cancer biomarkers, but their cellular origin and regulatory mechanisms remain unclear. Prostate samples from 70 men (30 prostate cancer, 40 benign) were analyzed by in situ hybridization for nine miRNAs identified via serum next-generation sequencing (NGS). Digital image analysis was used to quantify miRNAs in benign and malignant epithelial compartments and stromal cells. Five of the nine miRNAs were detected in prostate tissue. miR-550a, miR-4754, and miR-4326 exhibited an “epithelial down, stromal up” pattern accompanied by elevated serum levels; serum miR-550a correlated with high-grade tumor cells. miR-1246 progressively decreased from benign epithelium through Gleason grade 3 to grade 4 tumor cells, suggesting increased secretion by advanced tumor cells. However, extraprostatic contributions and assay cross-reactivity likely influenced its serum profile. miR-4632 exhibited distinct compartmental alterations, but methodological or extraprostatic factors appeared to affect serum detection. Overall, this integrative profiling reveals that not all miRNAs altered in serum from prostate cancer patients originate from prostate tissue, underscoring the value of comparative tissue/serum analyses. miR-550a, miR-1246, miR-4754, and miR-4326 emerge as promising biomarkers warranting further longitudinal evaluation.

Alzheimer's disease (AD) is a progressive neurodegenerative disorder, with mild cognitive impairment (MCI) as its critical prodromal stage. Identifying reliable biomarkers to distinguish AD from MCI is key for early intervention. This study explored miR-769-5p's potential as a diagnostic biomarker for AD and its mechanism, especially its links to oxidative stress and target gene CREB5. Serum miR-769-5p and CREB5 levels were measured via qRT-PCR. Cellular models used Aβ₄₂-treated SH-SY5Y cells, validated by confirming key AD pathologies including increased Tau phosphorylation. Functional/mechanistic assessments employed CCK-8, MDA, SOD, and dual-luciferase reporter assays. Clinically, multivariate logistic regression identified miR-769-5p as a strong independent protective factor (OR = 0.029, p < 0.001). miR-769-5p was downregulated in AD versus MCI and accurately distinguished them (AUC = 0.919). In Aβ-treated cells, miR-769-5p overexpression reduced Aβ-induced proliferation suppression and oxidative stress (lower MDA, higher SOD, all p < 0.01), while inhibition worsened damage. CREB5, a direct miR-769-5p target, was upregulated in AD. Functionally, rescue assays confirmed CREB5 overexpression reversed miR-769-5p mimic's protection. In conclusion, serum miR-769-5p is a promising biomarker for AD-MCI distinction and may protect against AD by targeting CREB5 to mitigate oxidative stress.

The diagnosis of lymphadenitis caused by nontuberculous mycobacteria (NTM) can be difficult, time consuming, and may need invasive investigations. The aim of this study was to analyze the use of a simple blood test and PPD stimulation in the diagnosis of NTM infections in non-BCG-vaccinated children and to determine the cut-off value for PPD response. For this purpose, the result of a modified Elispot IGRA assay and clinical data from 939 pediatric patients tested in 2011–2015 at the Helsinki University Hospital were reviewed retrospectively. Altogether, 45 children with NTM cervical lymphadenitis were identified. Fifteen of the cases were confirmed by culture and comprised the study group for the PPD cut-off value calculation. One hundred forty-one healthy, non-BCG-vaccinated children with recent contact with an MTB positive case comprised the control group. The cut-off value for PPD response for the diagnosis of NTM cervical lymphadenitis was 29 spots/million lymphocytes. The PPD antigen in the Elispot IGRA test can also be used in the evaluation of the BCG vaccination status. The PPD cut-off for BCG vaccination was 36 spots/million lymphocytes. In conclusion, the PPD stimulation response is a useful minimally invasive diagnostic method in the diagnosis of NTM cervical lymphadenitis in non-BCG-vaccinated children.

Although dexamethasone reduces mortality in patients with COVID-19, plasma glucose (PG) levels increase upon initiation. In a multicenter observational cohort of 530 adults, we estimated glycemic variability based on baseline HbA1c among patients with normoglycemia (N = 238), prediabetes (N = 159), unknown (N = 63), and known diabetes (N = 159). Glycemic variability, diabetic- and hyperglycemic events (≥ 11.1 and ≥ 16 mmol/L) were analyzed using a linear mixed model and competing risks analysis adjusted for confounders. Before dexamethasone, mean PG levels were similar in those with normoglycemia (6.5 mmol/L) and prediabetes (6.6 mmol/L), but higher in unknown (8.5 mmol/L) and known diabetes (9.9 mmol/L). After treatment, PG increased across all groups. Prediabetes showed a larger increase (1.5 mmol/L) than normoglycemia (0.7 mmol/L, p = 0.002), and known diabetes had the highest increase (2.4 mmol/L, p < 0.001), reaching an average of 12.6 mmol/L. All groups except prediabetes returned to baseline after dexamethasone. The cumulative incidence of diabetic events was 98% in known diabetes, 67% in unknown diabetes, 31% in prediabetes, and 8% in normoglycemia, with significant differences between groups (p < 0.001). We conclude that dexamethasone treatment increased average PG and caused frequent hyperglycemic events in patients with prediabetes, unknown, and known diabetes, while persistent PG elevation post-treatment occurred in prediabetes.

Comamonas aquatica, an emerging nosocomial pathogen, poses significant clinical challenges through biofilm-mediated antimicrobial resistance. This study investigated the efficacy of cuminaldehyde combined with tetracycline against C. aquatica biofilms using an integrated approach. In silico predictions (PASS online, SwissADME, PROTOX 3.0, OSIRIS) indicated that cuminaldehyde exhibited favorable oral bioavailability with acceptable toxicity profiles, while tetracycline showed limited oral absorption due to molecular size and polarity constraints. Experimentally, individual minimum inhibitory concentrations (MICs) were determined as 300 μg/mL for cuminaldehyde and 0.2 μg/mL for tetracycline. The fractional inhibitory concentration index (FICI) of 0.66 demonstrated additive interactions between the compounds (cuminaldehyde and tetracycline). The result indicated that the combinatorial application of compounds exhibited enhanced antimicrobial potential against the test organism. Furthermore, co-application of cuminaldehyde and tetracycline was found to show increased antibiofilm potential against the same organism. The result showed that the biofilm inhibition under the influence of the combinatorial application could be attributed to the enhancement of bacterial cell membrane permeability and accumulation of intracellular reactive oxygen species. In a nutshell, the findings of this study highlight a promising strategy of using combinatorial therapy involving cuminaldehyde-tetracycline for dealing with biofilm-associated infections caused by C. aquatica.

Bone and joint infections (BJIs) are debilitating conditions that reduce quality of life. Key features include interosseous bacteria, biofilm formation, suppurative inflammation, and osteolysis. Traditionally, bacteria-associated osteolysis has been attributed to RANKL-mediated osteoclast activation, based largely on in vitro and murine studies, but emerging evidence challenges this view. In this translational study, a porcine implant-associated osteomyelitis (IAO) model was combined with clinical fracture-related infection (FRI) data to investigate mechanisms of bone loss. In the IAO model, RANKL signaling was inhibited using Denosumab, yet pathological and radiographic osteolysis remained unchanged. Local RANKL mRNA expression and active osteoclast numbers also did not correlate with bone destruction. In FRI patients, mRNA in situ hybridization showed that MMP1 expression was higher in osteolytic cases compared to non-osteolytic ones, whereas RANKL expression did not differ. The technique revealed heterogeneous expression of MMP1 and RANKL, in contrast to uniform C3 expression. The number of active osteoclasts likewise did not correlate with osteolysis severity, and a patient with chronic osteomyelitis receiving Denosumab for osteoporosis still experienced extensive bone loss. These findings do not refute the RANKL–osteoclast pathway but indicate that bacteria-associated osteolysis is multifactorial, shaped by inflammatory and osteoimmunological interactions, proteolysis, neutrophil activity, and impaired osteogenesis.

Acinetobacter baumannii is a Gram-negative opportunistic pathogen linked to over 400,000 deaths annually, with International Clone 2 (IC2) recognised as a major multidrug-resistant (MDR) lineage. We analysed 668 IC2 genomes from BIGSdb, assessed completeness using usegalaxy-BUSCO, and annotated them with Prokka. Protein–protein interactions were mapped using STRING; interacting genes were identified using Cytoscape; functional enrichment was conducted using Enrichr and KEGG, retrieving protein from UniProt; and potential drug candidates were obtained from ChEMBL. Molecular docking was carried out using Autodock 4, and the complexes were visualised in Discovery studio. ADME profiling with SWISSADME and toxicity were assessed using MCULE and ProTOX-3.0. We identified polA as a major hub gene involved in DNA repair and essential for survival under genotoxic antibiotic stress, suggesting that it could impair bacterial adaptability and resistance mechanisms. Molecular docking using AutoDock Vina identified Nodulisporone and Nodulisporol from ChEMBL as potential inhibitors that also fulfilled Lipinski's criteria, with higher binding affinities for A. baumannii PolA than for human DNA polymerase lambda (PolL). This suggests that polA inhibition may potentiate existing antibiotics, reducing the dissemination of resistance and disrupting essential repair processes. Our bioinformatic and computational biology approach offers an effective strategy for developing novel treatments against multidrug-resistant pathogens.