Diabetic kidney disease (DKD), characterized by mesangial fibrosis and renal dysfunction, is a major microvascular complication of diabetes. Studies have shown that miRNAs are closely related to the progression of DKD. Therefore, in this study, we aimed to explore whether miR-1225-3p can regulate Smad ubiquitin regulatory factor 2 (SMURF2)-mediated carbohydrate response element binding protein (ChREBP) ubiquitination through Rho GTPase-activating protein 5 (ARHGAP5) to affect fibrosis in DKD.

DKD mice were established by intraperitoneally injecting streptozocin (STZ), and a DKD cell model was generated by culturing in media supplemented with 25 mmol/L glucose (high glucose, HG). StarBase was used to predict the target binding sites between miR-1225-3p and ARHGAP5, and a dual-luciferase reporter gene assay was used to verify this relationship. Western blotting, RT-qPCR, flow cytometry, immunoprecipitation, ELISAs, HE staining, and Masson staining were used to detect relevant indicators.

ARHGAP5 and SMURF2 expression was decreased, but ChREBP was highly expressed in the renal tissue of DKD mice and HG-induced mouse mesangial cells (MMCs). miR-1225-3p could target and regulate the transcription of ARHGAP5, and an association between ARHGAP5 and SMURF2 was revealed. miR-1225-3p facilitated fibrosis and oxidative stress in MCCs by inhibiting ARHGAP5. In addition, SMURF2 promoted the ubiquitination of HA-ChREBP, and miR-1225-3p facilitated fibrosis and oxidative stress by mediating the ARHGAP5/SMURF2-mediated ubiquitination of ChREBP in MCCs. Furthermore, the miR-1225-3p inhibitor inhibited fibrosis and inflammation in the renal tissues of DKD mice.

miR-1225-3p facilitates fibrosis and oxidative stress by mediating ARHGAP5/SMURF2-mediated ubiquitination of ChREBP.

Acute kidney injury (AKI) is a clinical syndrome with high morbidity and mortality. AKI has emerged as a significant global public health issue, particularly among hospitalized patients, with the highest incidence observed in those admitted to intensive care units (ICUs). However, early diagnosis of AKI remains challenging due to the limited sensitivity and specificity of conventional biomarkers, including serum creatinine and urine output. Recently, microRNAs (miRNAs) have garnered increasing interest for their potential in the early detection and management of AKI. Owing to their high stability, ease of quantification, well-characterized regulatory functions, and close association with key pathophysiological processes, miRNAs are considered promising diagnostic and therapeutic candidates. Nevertheless, the clinical utility of miRNAs remains limited by confounding factors such as co-infections, comorbidities, and medication use, which may lead to false-positive results. Challenges also persist regarding off-target effects and developing safe and efficient delivery systems. Furthermore, only a few studies have systematically characterized miRNA expression profiles in AKI, considering its heterogeneous etiologies and the dynamic nature of miRNA regulation. Interactions between miRNAs and between miRNAs and non-coding RNAs such as circular (circRNAs) and long non-coding RNAs (lncRNAs) warrant further investigation.

The eukaryotic initiation factor 5A2 gene (EIF5A2) is a highly conserved and multifunctional gene that significantly influences various cellular processes, including translation elongation, RNA binding, ribosome binding, protein binding and post-translational modifications. Overexpression of EIF5A2 is frequently observed in multiple cancers, where it functions as an oncoprotein. Additionally, EIF5A2 is implicated in drug resistance through the regulation of various molecular pathways. In the review, we describe the structure and functions of EIF5A2 in normal cells and its role in tumorigenesis. We also elucidate the molecular mechanisms associated with EIF5A2 in the context of tumorigenesis and drug resistance. We propose that the biological roles of EIF5A2 in regulating diverse cellular processes and tumorigenesis are clinically significant and warrant further investigation.

Endocrine therapy resistance (ETR) in breast cancer (BC) is a multicausal phenomenon with diverse alterations in the tumor cell interactome. Within these alterations, non-coding RNAs (ncRNAs) such as micro-RNAs (miRNAs) modulate the expression of tumor suppressor genes and proto-oncogenes, such as the ESR1 gene encoding estrogen receptor alpha (ERα). This work aims to review the effects of miRNAs targeting ERα mRNA and their mechanisms related to ETR in BC. A thorough review of the literature and an in silico study were carried out to elucidate the involvement of each miRNA, thus contributing to the understanding of ETR in BC.

Precision nutrition aims to tailor dietary interventions based on genetic and molecular profiles. MicroRNAs (miRNAs) and small interfering RNAs (siRNAs) are emerging as critical tools in precision obesity management. miRNAs serve as biomarkers for predicting dietary response and obesity risk, while siRNAs provide a targeted approach to silencing obesity-related genes. This review explores the mechanisms, applications, and potential of integrating miRNA and siRNA in personalized dietary strategies to combat obesity.

A comprehensive literature review was conducted using Boolean operations to identify studies on miRNAs, siRNAs, and their roles in precision nutrition. The review focused on molecular mechanisms, clinical applications, challenges, and future directions in integrating miRNA detection and siRNA therapy for obesity management.

miRNAs regulate gene expression related to lipid metabolism, adipogenesis, and insulin sensitivity, with miRNA-33 and miRNA-103/107 being notable examples. siRNAs offer precise gene silencing for targets like SREBP-1c and PPARγ, addressing metabolic pathways resistant to dietary interventions. The synergistic integration of miRNAs as biomarkers and siRNAs as therapeutic tools enhances the personalization and efficacy of obesity management.

The dual application of miRNAs and siRNAs in precision nutrition represents a transformative approach to obesity management. While challenges such as molecular stability and delivery systems persist, advancements in RNA technology and clinical research promise to revolutionize personalized dietary strategies. Future research should focus on large-scale trials and ethical considerations to ensure equitable and effective implementation.

While radiation-induced lung injury decreases quality of life and suppresses efficacy of radiotherapy, to date, the relationship between radiation-induced lung injury and repair remains unclear. Our previous studies revealed that TNFRSF10B-RIPK1/RIPK3-MLKL signaling induces necroptosis of alveolar epithelial cells and potentiates radiation-induced lung injury. We also found that microRNA-541-3p is differentially expressed in radiation-damaged lungs. The connection between microRNA-541-3p, TNFRSF10B signaling, and TGFβ1 signaling is also unclear.

This study was performed to explore the regulatory effects of microRNA-541-3p on TNFRSF10B and TGFβ1 signaling.

Mouse alveolar epithelial cells were transfected with a vector expressing microRNA-541-3p to regulate expression of target genes. Flow cytometry, polymerase chain reaction, and western blotting were used to analyze cell necroptosis, target gene expression, and target protein expression, respectively.

Overexpression of microRNA-541-3p positively regulated TNFRSF10B-RIPK1/RIPK3-MLKL signaling through Rac2 to induce cell necroptosis. MicroRNA-541-3p negatively regulates Rac2. MicroRNA-541-3p and Rac2 regulate the expression of Tgf-beta1 and its encoded proteins.

The Rac2 gene synchronously regulates TNFRSF10B-RIPK1/RIPK3-MLKL and TGFβ1 signaling. MicroRNA-541-3P/Rac2 act as mediators of radiation damage and repair signaling.

Severe corneal injuries often result in corneal scarring, leading to visual impairment and corneal blindness. Currently, there is a lack of effective anti-corneal fibrosis drugs in clinical practice. MicroRNA-based therapies hold significant potential in combating fibrosis. However, the barrier function of the cornea and the fluid environment of the ocular surface reduce drug permeability and bioavailability, presenting significant challenges for local drug application. This study employs microfluidic technology to encapsulate miRNA29b in lipid nanoparticles (LNP) to create an LNP-miRNA29b delivery system (LNP-mir29b) for treating corneal mechanical injuries. In vitro experiments show that LNP-mir29b significantly inhibits the expression of α-smooth muscle actin (α-SMA) in an induced corneal stromal cell fibrosis model. In vivo experiments using rabbit corneal mechanical injury models indicate that LNP-mir29b effectively reduces fibrosis in the corneal stroma, promotes organized rearrangement of stromal collagen fibers, and decreases the expression of fibrosis-related genes, including Col1A2, Col3A1, Fn, and α-SMA. Additionally, LNP-mir29b accelerates the migration of corneal epithelial cells, promotes wound healing of the epithelium, restores the structural integrity of the corneal epithelium. The LNP system proposed in this study offers a novel approach with anti-fibrotic functionality, providing a new strategy for reducing scarring during the corneal injury repair process.

miRNAs are critical for different disease development processes, including cell growth, signaling, apoptosis, cancer and neurodegenerative diseases. It has been shown that altered miRNA levels are associated with reactive oxygen species (ROS) formation and mitochondrial dysfunction. While mitochondrial dysfunction and ROS formation occur in many neurodegenerative diseases, such as Alzheimer's disease, Parkinson's disease, and multiple sclerosis, amyotrophic lateral sclerosis, miRNAs have the potential to be diagnostic biomarkers and therapeutic targets with a high degree of specificity, which is highly relevant in neurodegenerative pathologies.This paper gives a general summary of the current expression of miRNAs in neurodegenerative diseases, including miRNAs up-regulated or down-regulated in a variety of diseases, as well as the associated factors of influence. miRNAs are more like a double-edged sword, their multi-targeted role has brought light to many diseases for which there are currently no clear therapeutic options, but at the same time, their low specificity and possible side effects on the whole body should not be ignored, therefore However, at the same time, its low specificity and possible side effects on the whole body should not be ignored, therefore, more attention should be paid to the development of miRNA therapy in terms of its high efficiency, the use of carriers, and the clarification of side effects.

MicroRNAs (miRNAs) have emerged as promising tools for diagnosis and treatment in numerous pathophysiological processes, including cardiovascular diseases (CVD). In this context, acute myocardial infarction (AMI) is one of the leading causes of death by CVD worldwide. In this sense, physical exercise (PE) is considered a non-pharmacological strategy to reduce the complex alterations in AMI. This study is an integrative review of the literature to explore the effects of PE on the cardiomyocyte post-AMI, including an understanding of the mechanisms by which the PE acts on the miRNAs expression. A review was performed on PubMed, Scopus, and Web of Science. After the searches, all records were imported into the Mendeley software, and duplicate articles were removed. The year of publication of the papers was not limited. 19 studies were performed on animal models, 10 in experimental models using rats, and 08 in models with mice and only one study was carried out on patients with AMI. The results showed the potential use of miRNAs as diagnostic tools and attractive biomarkers for treating AMI. In addition, PE can regulate miRNAs expression in the myocardial cell, promotes apoptosis resistance, autophagy regulation, lower cardiac fibrosis and cardiac hypertrophy, and higher angiogenesis through the signaling of miRNAs. The main microRNAs mitigating the deleterious effects of AMI and modulated by PE were miRNA-222, miRNA-1192, miRNA-146, and miRNA-126. PE modulates the expression of specific miRNAs that support cardiac function, promoting cardioprotective effects or facilitating cardiac recovery post-AMI.

miR-135b, a microRNA, is consistently up-regulated in various cancer tissues and cells, promoting cancer progression. By inhibiting one or more target genes, miR-135b regulates phenotypes such as cancer growth, apoptosis, migration, invasion, drug resistance, and angiogenesis, establishing it as a critical driver of cancer progression. Additionally, miR-135b is regulated by various oncogenes and therapeutic drugs, highlighting its complexity and therapeutic potential. Significant progress has been made in understanding miR-135b's impact on cancer cell behavior, establishing it as a promising biomarker for cancer diagnosis and prognosis, as well as a potential target for future cancer therapies. However, despite the extensive research on this topic, there has been no comprehensive review summarizing its role and mechanisms across different cancer types. This review aims to provide a detailed overview of the biological characteristics of miR-135b, its regulatory targets, upstream signaling pathways, and its therapeutic potential, including its influence on cancer chemoresistance. The review also addresses key controversies surrounding miR-135b in cancer research, aiming to deepen the understanding of its role, promote the transformation of its clinical application, and provide a theoretical foundation for developing more effective cancer treatment strategies.

The present study was conducted with the objective ascertaining the clinical implication of microRNA-133a-3p (miR-133a-3p) for urinary incontinence (UI) and rehabilitation effects in prostate cancer after radical prostatectomy.

The measurements of miR-133a-3p in urethral tissue samples from prostate cancer patients after radical prostatectomy were carried out via quantitative real-time polymerase chain reaction (qRT-PCR) detection. Receiver operation characteristic (ROC) curve and logistic regression analysis were employed for evaluating the predictive significance of miR-133a-3p for the early UI of prostate cancer patients with radical prostatectomy. Bioinformatics tools were employed for mining the miR-133a-3p possible genes.

An obvious reduction of miR-133a-3p was detected in patients with UI compared with those with urinary continence (UC) (P < 0.001), demonstrating a high diagnostic capacity for patients with UI. Moreover, miR-133a-3p could be an independent predictive index for the early UI in patients with prostate cancer after radical prostatectomy (P < 0.001). Additionally, urine miR-133a-3p was notably increased in the UI patients after rehabilitation (P < 0.001). MiR-133a-3p largely concentered on the muscle-related diseases pathways using bioinformatics tools.

MiR-133a-3p was a promising indicator for predicting early UI in patients with prostate cancer after radical prostatectomy.

Not applicable.

Sinonasal inverted papilloma (SNIP) is a benign epithelial neoplasm of the Schneiderian membrane, known for its locally aggressive behavior, high recurrence rates, and potential for malignant transformation into sinonasal squamous cell carcinoma (SNSCC). Emerging evidence emphasizes the role of microRNAs (miRNAs) in the pathogenesis, progression, and clinical management of SNIP. These small non-coding RNAs regulate key cellular pathways, particularly the PTEN/PI3K/AKT axis, which governs tumor growth, apoptosis resistance, and chemoresistance. Among the miRNAs studied, miR-296-3p, miR-214-3p, and the miR-449 cluster show significant dysregulation. miR-296-3p is upregulated in SNSCC, promoting oncogenesis by inhibiting PTEN and activating the PI3K/Akt pathway. Conversely, miR-214-3p is downregulated in SNIP and correlates with advanced disease and increased recurrence, identifying it as a potential diagnostic and prognostic biomarker. The miR-449 cluster, with known tumor-suppressive properties, is progressively downregulated during malignant transformation, highlighting its role in maintaining epithelial structure. Despite their promise, clinical application of miRNA-based diagnostics and therapies faces challenges such as delivery optimization, specificity, and off-target effects. Nonetheless, the noninvasive detection of circulating miRNAs in bodily fluids offers a compelling approach for future diagnostic tools and patient monitoring. This review highlights the transformative potential of miRNA research in advancing SNIP diagnosis and treatment. By integrating molecular insights into clinical practice, miRNA-based strategies could pave the way for more personalized interventions, ultimately reducing recurrence rates and preventing malignant transformation.

The field of RNA research has experienced significant changes and is now at the forefront of contemporary drug development. This narrative overview explores the scientific developments and historical turning points in RNA research, emphasising the field's critical significance in the development of novel therapeutics. Important discoveries like antisense oligonucleotides (ASOs), mRNA therapies, and RNA interference (RNAi) have created novel treatment options that can be targeted, such as the ground-breaking mRNA vaccinations against COVID-19. Advances in high-throughput sequencing, single-cell RNA sequencing, and epitranscriptomics have further unravelled the complexity of RNA biology, shedding light on the intricacies of gene regulation and cellular diversity. The integration of computational tools and bioinformatics has propelled the identification of RNA-based biomarkers and the development of RNA therapeutics. Despite significant progress, challenges such as RNA stability, delivery, and off-target effects persist, necessitating continuous innovation and ethical considerations. This review provides a critical insight into the current state and prospects of RNA research, emphasising its transformative potential in drug discovery. By examining the interplay between technological advancements and therapeutic applications, we underscore the promising horizon for RNA-based interventions in treating a myriad of diseases, marking a new era in precision medicine.

MicroRNAs (miRNAs) are small non-coding RNAs that regulate various biological processes, including cell differentiation. Despite their potential, their role in promoting neuronal differentiation by targeting neuronal genes and modulating signaling pathways is poorly understood. In this study, we aimed to elucidate the functions of miR-937-3p-, miR-4536-3p-, and miR-4650-5p-inhibitors in the neuronal differentiation of SH-SY5Y cells. We also aimed to determine the underlying mechanisms via qPCR, luciferase assay, immunocytochemistry, and western blotting analysis. Our findings confirmed that miRNAs participated in neuronal differentiation and regulated the Wnt/MAPK signaling pathway. Specifically, we identified Netrin1 (NTN1), Drebrin1 (DBN1), and Netrin-G1 (NTNG1) as target genes of miR-937-3p, miR-4536-3p, and miR-4650-5p, respectively. The treatment with the miRNA inhibitors increased the expression levels of neuronal markers such as TUBB3, NEFH, NEFM, NEFL, and MAP2. It also enhanced the protein expression levels of Wnt and MAPK signaling. Therefore, the inhibitors of miR-937-3p, miR-4536-3p, and miR-4650-5p could promote neuronal differentiation by targeting neuronal genes and activating the Wnt/MAPK signaling pathway.

There is no diagnostic test for primary immune thrombocytopenia (ITP). Certain microRNAs have shown to have diagnostic potential in ITP. We validated 12 microRNAs identified from two previous studies to find a diagnostic biomarker. The study included two ITP cohorts (n = 61) and healthy controls (n = 28). The first ITP cohort involved 24 patients from the Prolong study, patients with newly diagnosed/persistent ITP (<1 year) treated with corticosteroids ± IVIG but relapsed/failed to respond. The second cohort comprised 37 patients from ITP biobank, Østfold Hospital, Norway, patients had different disease stages and therapies. Twelve microRNAs were measured: miR-199a-5p, miR-33a-5p, miR-195-5p, miR-130a-3p, miR-144-3p, miR-146a-5p, miR-222-3p, miR-374b-5p, miR-486-5p, miR-1341-5p, miR-766-3p and miR-409-3p. miR-199a-5p, miR-33a-5p, miR-374b-5p, miR-146a-5p and miR-409-3p were expressed differentially in the entire ITP cohort compared to controls; of those only miR-199a-5p showed good discriminative ability between ITP and controls with area under the curve (AUC) of 0.718 (95% CI: 0.599-0.836). In the Prolong cohort (ITP < 1 year), miR-199a-5p and miR-374b-5p showed very good discriminative ability between ITP and controls with AUC of 0.824 (0.708-0.940) and 0.806 (0.688-0.924) respectively. This study confirmed that miR-199a-5p has good discriminative ability between primary ITP and healthy controls, thus may be a diagnostic biomarker of ITP.

Epithelial ovarian cancer (EOC) is the most common subtype of ovarian cancer and is a highly recurrent and lethal malignancy of the female reproductive system. Accumulating evidence has indicated that long non-coding RNAs (lncRNAs) play pivotal roles in tumorigenesis. Long intergenic non-protein coding RNA 704 (LINC00704) has been recognized as an oncogenic lncRNA in several malignancies. However, its specific role in EOC remains unclear. In this study, RT-qPCR revealed the upregulation of LINC00704 in EOC tissues and cell lines. CCK-8 and EdU assays showed that LINC00704 knockdown inhibited EOC cell proliferation. Flow cytometry analysis demonstrated that LINC00704 silencing induced EOC cell cycle arrest and apoptosis. Transwell assays indicated the inhibitory effects of LINC00704 silencing on EOC cell migration and invasion. Mechanistically, bioinformatics analysis, RNA pull-down, luciferase reporter and RNA immunoprecipitation assays revealed that LINC00704 upregulated SLC44A1 expression by competitively binding to miR-323a-3p. Moreover, rescue experiments showed that SLC44A1 overexpression could reverse LINC00704 silencing-mediated effects on the malignant behaviors of EOC cells. In conclusion, LINC00704 promotes EOC cell aggressiveness in vitro by regulating the miR-323a-3p/SLC44A1 axis.

The high mortality rate of ARDS is closely related to pulmonary fibrosis (PLF), and the degree of pulmonary fibrosis affects the prognosis of ARDS. Numerous studies indicated the abnormal expression of miRNAs in the pathogenesis of various lung diseases, such as ARDS and PLF. This study aimed to explore the expression of serum miR-6515-5p and its clinical performance in ARDS complicated with PLF.

RT-qPCR analysis was employed to measure miR-6515-5p levels within the serum specimens from ARDS patients who either had PLF or did not. Receiver Operating Characteristics (ROC) curve was conducted to evaluate the diagnostic value of miR-6515-5p. To analyze the risk factors linked with the development of PLF in ARDS patients, logistic regression analysis was conducted. Kaplan-Meier curve was conducted to assess the prognostic value of miR-6515-5p in predicting the outcome of ARDS patients with PLF. Multivariate COX regression analysis was performed to identify the PLF-related risk factors associated with outcomes.

Serum miR-6515-5p was downregulated in ARDS patients, especially in patients suffering from PLF. miR-6515-5p expression can distinguish ARDS patients from healthy individuals and related to the occurrence of PLF. Most patients with low miR-6515-5p expression had low FVC and DLCO, as well as high Murray score and APACHE II score. Moreover, miR-6515-5p expression has a certain high value in differentiating ARDS patients with PLF from those without PLF. In addition, miR-6515-5p may be a prognostic marker in ARDS patients suffering from PLF.

These data identified the abnormal expression of miR-6515-5p in ARDS and PLF, and implied a potential clinical early diagnostic and prognostic marker in ARDS suffering from PLF.

Depression, particularly major depressive disorder (MDD), is a debilitating neuropsychiatric condition characterized by high disability rates, primarily driven by chronic stress and genetic predispositions. Emerging evidence highlights the critical role of microRNAs (miRNAs) in the pathogenesis of depression, with plasma-derived small extracellular vesicles (sEVs) emerging as promising biomarkers. In this study, we collected peripheral blood plasma samples from patients diagnosed with MDD, as assessed by the Hamilton Depression Rating scale, alongside healthy individuals serving as controls. Plasma-derived sEVs were isolated via ultracentrifugation, followed by high-throughput sequencing of miRNAs encapsulated within sEVs, and finally image acquisition and differential expression analysis. Our results revealed a significant elevation of miR-182-5p in plasma-derived sEVs from MDD patients compared to healthy controls, a finding further validated in chronic mild stress (CMS) models. Further analysis suggested that miRNAs encapsulated within sEVs may influence depression onset and progression by modulating hypothalamic-pituitary-adrenal (HPA) axis activity. These findings underscore the potential of miRNAs and their target genes as novel biomarkers, offering improved diagnostic accuracy and therapeutic efficacy for MDD.

This review explores the dual role of miR-195 in cancer, acting as both a tumor suppressor and, in specific contexts, a tumor promoter. It highlights its molecular mechanisms, focusing on key signaling pathways such as Wnt-1/β-catenin, VEGF/VEGFR, and PI3K/AKT/mTOR, as well as its involvement in competitive gene regulation. The clinical potential of miR-195 in cancer screening, diagnosis, prognosis, and therapy is examined, particularly its ability to enhance therapeutic efficacy and reduce recurrence risk when combined with chemotherapy or immunotherapy. Despite these promising aspects, challenges such as precise regulation, efficient delivery systems, and clinical translation remain. Future research should prioritize advancing miR-195's integration into personalized medicine, immunotherapy, and novel delivery technologies, aiming to establish it as a reliable biomarker and therapeutic target for improved cancer care.

Hemorrhagic neurovascular diseases, with high mortality and poor outcomes, urge novel biomarker discovery and therapeutic targets. Micro-ribonucleic acids (miRNAs) are potent post-transcriptional regulators of gene expression. They have been studied in association with disease states and implicated in mechanistic gene interactions in various pathologies. Their presence and stability in circulating fluids also suggest a role as biomarkers. This review summarizes the current state of knowledge about miRNAs in the context of cerebral cavernous malformations (CCMs), a disease involving cerebrovascular dysmorphism and hemorrhage, with known genetic underpinnings. We also review common and distinct miRNAs of CCM compared to other diseases with brain vascular dysmorphism and hemorrhage. A systematic search, following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guideline, queried all peer-reviewed articles published in English as of January 2025 and reported miRNAs associated with four hemorrhagic neurovascular diseases: CCM, arteriovenous malformations, moyamoya disease, and intracerebral hemorrhage. The PubMed systematic search retrieved 154 articles that met the inclusion criteria, reporting a total of 267 unique miRNAs identified in the literature on these four hemorrhagic neurovascular diseases. Of these 267 miRNAs, 164 were identified in preclinical studies, while 159 were identified in human subjects. Seventeen miRNAs were common to CCM and other hemorrhagic diseases. Common and unique disease-associated miRNAs in this systematic review motivate novel mechanistic hypotheses and have potential applications in diagnostic, predictive, prognostic, and therapeutic contexts of use. Much of current research can be considered hypothesis-generating, reflecting association rather than causation. Future areas of mechanistic investigation are proposed alongside approaches to analytic and clinical validations of contexts of use for biomarkers.

MicroRNAs (miRNAs) are small RNA molecules that do not have coding functions but play essential roles in various biological processes. In lung cancer, miRNAs affect the processes of tumor initiation, progression, metastasis, and resistance to treatment by regulating gene expression. Tumor-suppressive miRNAs inhibit oncogenic pathways, while oncogenic miRNAs, known as oncomiRs, promote malignant transformation and tumor growth. These dual roles position miRNAs as critical players in lung cancer biology. Studies in recent years have shown the significant potential of miRNAs as both prognostic and diagnostic biomarkers. Circulating miRNAs in plasma or sputum demonstrate specificity and sensitivity in detecting early-stage lung cancer. Liquid biopsy-based miRNA panels distinguish malignant from benign lesions, and specific miRNA expression patterns correlate with disease progression, response to treatment, and overall survival. Therapeutically, miRNAs hold promise for targeted interventions. Strategies such as miRNA replacement therapy using mimics for tumor-suppressive miRNAs and inhibition of oncomiRs with antagomiRs or miRNA sponges have shown preclinical success. Key miRNAs, including the let-7 family, miR-34a, and miR-21, are under investigation for their therapeutic potential. It should be emphasized that delivery difficulties, side effects, and limited stability of therapeutic miRNA molecules remain obstacles to their clinical use. This article examines the roles of miRNAs in lung cancer by indicating their mechanisms of action, diagnostic significance, and therapeutic potential. By addressing current limitations, miRNA-based approaches could revolutionize lung cancer management, offering precise, personalized, and minimally invasive solutions for diagnosis and treatment.

Early screening methods for gastric cancer (GC) are lacking; therefore, the disease often progresses to an advanced stage when patients first start to exhibit typical symptoms. Endoscopy and pathological biopsy remain the primary diagnostic approaches, but they are invasive and not yet widely applicable for early population screening. miRNA is a highly conserved type of RNA that exists stably in plasma. Dysfunction of miRNA is linked to tumorigenesis and progression, indicating that individual miRNAs or combinations of multiple miRNAs may serve as potential biomarkers.

To identify effective plasma miRNA biomarkers and investigate the clinical value of combining multiple miRNAs for early detection of GC.

Plasma samples from multiple centres were collected. Differentially expressed genes among healthy controls, early-stage GC patients, and advanced-stage GC patients were identified through small RNA sequencing (sRNA-seq) and validated via real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR). A Wilcoxon signed-rank test was used to investigate the differences in miRNAs. Sequencing datasets of GC serum samples were retrieved from the Gene Expression Omnibus (GEO), ArrayExpress, and The Cancer Genome Atlas databases, and a multilayer perceptron-artificial neural network (MLP-ANN) model was constructed for the key risk miRNAs. The pROC package was used to assess the discriminatory efficacy of the model.

Plasma samples of 107 normal, 71 early GC and 97 advanced GC patients were obtained from three centres, and serum samples of 8443 normal and 1583 GC patients were obtained from the GEO database. The sRNA-seq and RT-qPCR experiments revealed that miR-452-5p, miR-5010-5p, miR-27b-5p, miR-5189-5p, miR-552-5p and miR-199b-5p were significantly increased in early GC patients compared with healthy controls and in advanced GC patients compared with early GC patients (P < 0.05). An MLP-ANN model was constructed for the six key miRNAs. The area under the curve (AUC) within the training cohort was 0.983 [95% confidence interval (CI): 0.980-0.986]. In the two validation cohorts, the AUCs were 0.995 (95%CI: 0.987 to nearly 1.000) and 0.979 (95%CI: 0.972-0.986), respectively.

Potential miRNA biomarkers, including miR-452-5p, miR-5010-5p, miR-27b-5p, miR-5189-5p, miR-552-5p and miR-199b-5p, were identified. A GC classifier based on these miRNAs was developed, benefiting early detection and population screening.

This study aimed to investigate the regulatory effect of linc00963 on postmenopausal osteoporosis and the potential molecular mechanisms.

Taking MC3T3-E1 cells as the study object, a cell cycle assay was used to evaluate the effect of linc00963 on cell proliferation. mRNA levels of Runx2, OCN, collagenia-1, OPG, RANKL and RANK were detected. Dual luciferase reporter assay verified the targeting relationship between linc00963 and miR-506-3p. A postmenopausal osteoporosis rat model was established after ovariectomy in 32 Sprague-Dawley rats. The rats were divided into sham group, OVX group, linc00963 overexpression group, and blank plasmid group. The bone mineral density (BMD) of the rat femur was measured by X-ray bone densitometer. Serum linc00963 expression in rat was detected by RT-qPCR. The protein expression of ALP, and BGP in the serum of rats was detected by ELISA.

Cell studies have shown that linc00963 alleviates postmenopausal osteoporosis by down-regulating the expression of miR-506-3p. Animal studies showed that compared with the sham group, the serum linc00963 level, BMD, serum Ca, P, LEP, SOD, and OPG levels in the OVX group were significantly decreased, while the levels of body weight, ALP, BGP, IL-6, IL-13, RANKL, and RANK were significantly increased. Compared with the OVX group, the use of linc00963 overexpression plasmid can significantly improve the above indexes and play a corresponding therapeutic effect on menopausal osteoporosis rats.

Linc00963 is involved in the pathogenesis of postmenopausal osteoporosis by up-regulating miR-506-3p and activating the OPG/RANKL/RANK pathway. Linc00963 is expected to be a potential therapeutic target for postmenopausal osteoporosis.

Background/Objectives: Ischemic stroke remains a leading cause of death and disability worldwide. Despite significant progress in reperfusion therapy, the optimal ischemic stroke management strategy has not been developed. Recent studies demonstrate that microRNA may play an essential role in the pathophysiology of ischemic stroke and its possible potential to be a treatment target point. The proposed systematic review aimed to report the relationship between IS's clinical severity and miRNA expression. Secondary outcomes included infarct volume, systemic inflammatory markers, and prognosis, as well as additional features such as stroke subtype, comorbidity, and risk of subsequent stroke in correlation to miRNA expression. Methods: We have performed a systematic search of database resources according to PRISMA statement guidelines. Twenty-seven studies on a total number of 3906 patients were assessed as suitable for the present SR. Included studies analyzed the expression of 30 different miRNA fragments. Results: After investigating available data, we have identified a set of possible miRNA fragment candidates that may be used in stroke diagnostics and have the potential to be a base for the development of future treatment protocols. Conclusions: Studies included in the presented SR indicate that miRNA expression may be significantly associated with clinical severity, infarct volume, and inflammation in ischemic stroke. More prospective, properly designed protocols with consistent methods of miRNA testing and optimized clinical assessment are needed to confirm the role of miRNA expression in the course of a stroke.

The significant role of microRNAs (miRNAs) in the regulation of pathological ocular neovascularization makes them both biomarkers and therapeutic targets in vascular eye diseases. However, their acute and sensitive detection and regulation continue to be a challenge. An integrative DNA probe (detection probe) was developed to enable the single-step detection of miRNAs with high sensitivity and specificity. This probe combines the functions of specific target recognition, the DNAzyme unit-based signal reaction, and the DNA polymerase/endonuclease-assisted signal cycle. In particular, a single-stranded sequence that engages with the DNAzyme unit of the s2 sequence can identify the target miRNA and liberate the DNAzyme unit to facilitate the signal reaction with the help of metal ions. In addition, the target recycling process and signal cycle were initiated by the DNA polymerase/endonuclease-assisted chain extension and displacement process, resulting in a low limit of detection of 4.56 fM. In addition, the method demonstrated a high level of septicity for the detection of target miRNAs and was capable of distinguishing interfering miRNAs with a one-base mismatch. Our research has shown that the integrative DNAzyme nanomachine is a promising biosensor for the sensitive detection and regulation of miRNA in a single step. It is anticipated that this technology will be used in the early diagnosis and therapy of vascular eye diseases.

Simple yet specific miRNA detection remains an enormous challenge due to its low abundance in samples and the high similarity among homologous miRNAs. Here, we propose a novel fluorescent approach for miRNA detection with greatly elevated accuracy by utilizing exonuclease-iii (Exo-iii) assisted twice target recognition. The proposed method involves a "Sensing probe" engineered with two loop sections to facilitate dual target miRNA recognition. The collaboration between Exo-iii and miRNA initiates target recycling for signal amplification, resulting in the formation of complete DNAzyme. The intact DNAzyme connects with the "Signal probe" and creates a nicking site within its loop region. The fluorescence signal of the "Signal probe" reemerges, correlating with the quantity of miRNA in the sensing system. The suggested technique demonstrates great selectivity for target miRNA and can readily differentiate sequences with a one-base mismatch, based on dual-target identification. Furthermore, the Exo-iii-assisted signal recycling imparts the approach with great sensitivity and a low detection limit of 548 aM. This method has the potential to be a robust alternative for the detection of miRNAs in real samples due to its high accuracy, simplicity, and resistance to potential fluorescence interferences.

Macropinocytosis is a nonselective form of endocytosis that allows cancer cells to largely take up the extracellular fluid and its contents, including nutrients, growth factors, etc. We first elaborate meticulously on the process of macropinocytosis. Only by thoroughly understanding this entire process can we devise targeted strategies against it. We then focus on the central role of the MTOR (mechanistic target of rapamycin kinase) complex 1 (MTORC1) in regulating macropinocytosis, highlighting its significance as a key signaling hub where various pathways converge to control nutrient uptake and metabolic processes. The article covers a comprehensive analysis of the literature on the molecular mechanisms governing macropinocytosis, including the initiation, maturation, and recycling of macropinosomes, with an emphasis on how these processes are hijacked by cancer cells to sustain their growth. Key discussions include the potential therapeutic strategies targeting macropinocytosis, such as enhancing drug delivery via this pathway, inhibiting macropinocytosis to starve cancer cells, blocking the degradation and recycling of macropinosomes, and inducing methuosis - a form of cell death triggered by excessive macropinocytosis. Targeting macropinocytosis represents a novel and innovative approach that could significantly advance the treatment of cancers that rely on this pathway for survival. Through continuous research and innovation, we look forward to developing more effective and safer anti-cancer therapies that will bring new hope to patients.Abbreviation: AMPK: AMP-activated protein kinase; ASOs: antisense oligonucleotides; CAD: carbamoyl-phosphate synthetase 2, aspartate transcarbamylase, and dihydroorotase; DC: dendritic cell; EGF: epidermal growth factor; EGFR: epidermal growth factor receptor; ERBB2: erb-b2 receptor tyrosine kinase 2; ESCRT: endosomal sorting complex required for transport; GAP: GTPase-activating protein; GEF: guanine nucleotide exchange factor; GRB2: growth factor receptor bound protein 2; LPP: lipopolyplex; MTOR: mechanistic target of rapamycin kinase; MTORC1: mechanistic target of rapamycin kinase complex 1; MTORC2: mechanistic target of rapamycin kinase complex 2; NSCLC: non-small cell lung cancer; PADC: pancreatic ductal adenocarcinoma; PDPK1: 3-phosphoinositide dependent protein kinase 1; PI3K: phosphoinositide 3-kinase; PIK3C3: phosphatidylinositol 3-kinase catalytic subunit type 3; PtdIns(3,4,5)P3: phosphatidylinositol-(3,4,5)-trisphosphate; PtdIns(4,5)P2: phosphatidylinositol-(4,5)-bisphosphate; PTT: photothermal therapies; RAC1: Rac family small GTPase 1; RPS6: ribosomal protein S6; RPS6KB1: ribosomal protein S6 kinase B1; RTKs: receptor tyrosine kinases; SREBF: sterol regulatory element binding transcription factor; TFEB: transcription factor EB; TNBC: triple-negative breast cancer; TSC2: TSC complex subunit 2; ULK1: unc-51 like autophagy activating kinase 1; UPS: ubiquitin-proteasome system.

β-elemene has a variety of anti-inflammatory, antioxidant, and antitumor effects. Currently, the influence of β-elemene on adrenocortical carcinoma (ACC) malignant progression and action mechanism remains unclear. This research aims to explore the influence and action mechanism of β-elemene on ACC progression. The impacts of β-elemene on ACC cell viability, proliferation, migration, and apoptosis were investigated through CCK-8 assay, clone formation assay, Transwell experiment, Wound healing assay, and flow cytometry. The miR-486-3p expression was analyzed utilizing RT-qPCR. According to different databases, neuronal pentraxin 1 (NPTX1) is the predicted downstream target gene of miR-486-3p. Western blot and RT-qPCR were utilized to examine NPTX1 expression. Silencing miR-486-3p or Overexpression NPTX1 in ACC cells further explored whether β-elemene affects ACC cells by regulating miR-486-3p/NPTX1. Finally, a subcutaneous graft tumor model was constructed to investigate how β-elemene may impact tumor growth in vivo. β-elemene decreased the cell viability, hindered cell proliferation and migration capacity, and induced apoptosis of ACC cells. miR-486-3p level in ACC cells was notably reduced in comparison to normal cells, but treatment with β-elemene markedly increased miR-486-3p expression. Additionally, ACC cells showed high level of NPTX1, while miR-486-3p targeted negative regulation of NPTX1. Overexpression miR-486-3p hindered the malignant progression of ACC cells, whereas overexpression NPTX1 reversed the impact of overexpression miR-486-3p. Silencing miR-486-3p or overexpression NPTX1 both attenuated the suppressive influence of β-elemene on the malignant behavior of ACC cells. Additionally, tumor growth was suppressed and apoptosis was induced in tumor cells in vivo by β-elemene. In conclusion, β-elemene reduces ACC cell viability, hinders proliferation and migration, and induces apoptosis through the miR-486-3p/NPTX1 axis.

Primary Sjögren's syndrome (pSS) is a chronic autoimmune disease characterized by inflammation and tissue destruction of the salivary and lacrimal glands, leading to sicca symptoms. Dysregulation of microRNAs (miRNAs), key post-transcriptional regulators, has been implicated in pSS, but their role in conjunctival epithelial cells (CECs) remains unclear. This study aimed to identify altered miRNA expression patterns in CEC from patients with pSS and their potential involvement in pSS pathogenesis.

CEC samples were collected from six patients with pSS and six healthy controls (HCs) using nylon-tipped swabs. The miRNA expression was profiled using the NanoString nCounter system with minimal RNA input. Differentially expressed (DE) miRNAs were identified via ROSALIND software, and bioinformatics tools (miRNet and miRTargetLink) were applied to construct miRNA-centric networks, predict target genes, and perform pathway enrichment analysis.

We identified 11 DE miRNAs in patients with pSS compared with the HCs. Key miRNAs, including hsa-miR-548j-3p and hsa-miR-219b-3p, are central to immune and inflammatory regulation pathways. Pathway enrichment analysis highlighted their involvement in processes such as immune cell regulation, inflammatory signaling, and glandular damage. Dysregulated miRNAs modulate key targets, like TNFAIP3, IL6R, IFNAR1, IL7, and ICOSLG, suggesting their potential role in pSS pathogenesis.

This study underscores the potential of miRNAs as biomarkers and therapeutic targets in pSS-associated dry eye disease. Despite limitations like small sample size and reliance on in silico predictions, our findings provide valuable insights into miRNA-mediated regulation of immune responses and inflammation, paving the way for future diagnostic and therapeutic advancements.

The aim of this study was to explore the bibliometric analysis of nanomaterials-based therapies for hepatocellular carcinoma as a means of assessing the current state of development and future trends in the field.

Literature on hepatocellular carcinoma and nanomedicine interactions was searched from the core database of the Web of Science and bibliometric and visualisation analyses were performed using VOSviewer, CiteSpace and GraphPad Prism data analysis software. We focused on important keywords, countries, authors, affiliations, journals, and literature in the field of nanomaterials for HCC.

The search resulted in the finalization of 421 documents. The search resulted in the finalization of 421 documents. From 2008 to 2023, nanomedicine research in HCC has developed rapidly, and the number of published papers has steadily increased, increasing by about 2300%. There are currently 57 countries involved in research in this area. Among them, The USA had the strongest international cooperation network and cooperated most closely with China. Gene delivery and carbon nanotubes were early keywords, immunotherapy and nanocarriers are recent research hotspots. It is important that the selection of nanocarriers and drug delivery have become the core trends driving the development of hepatocellular carcinoma.

The combination of nanomaterials with traditional imaging techniques such as MRI can improve the early diagnosis rate of HCC. Nanomaterials can achieve precise targeting of cancer cells by encapsulating drugs, loading bioactive molecules or modifying specific targeting ligands, thus significantly improving drug efficacy and effectively reducing adverse reactions in therapy.

The diagnosis and monitoring of eosinophilic esophagitis (EoE), a common pediatric pathology, typically involves invasive procedures such as an upper endoscopy with biopsies, imposing a significant burden on patients and healthcare systems. We aimed to assess miR-21-5p and miR-223-3p levels in pediatric EoE patients and evaluate their as potential non-invasive biomarkers of disease activity and response to treatments. We enrolled 13 children with EoE and 8 controls. Plasma and esophageal mucosa samples from patients were collected at diagnosis and after 8-10 weeks of therapy and compared with control samples. After microRNA(miRNA) extraction, the levels of miR-21-5p and miR-223-3p and their relevant target genes were analyzed. Bioinformatic analysis was used to identify the predicted target genes and pathways that are potentially relevant for disease pathophysiology. Plasma levels of miR-21-5p and miR-223-3p were significantly higher in EoE patients than in the controls, reflecting their levels in esophageal mucosa. The target genes of these miRNAs are involved in key signaling pathways (MAPK, Ras, and FoxO), relevant for EoE pathophysiology. Among these, STAT3 (Signal Transducer and Activator of Transcription 3) and PTEN (Phosphatase and Tensin Homolog), which are significantly downregulated in patient esophageal mucosa, are implicated in eosinophilic gastroenteropathies and autoimmune diseases. Following therapy (proton pump inhibitors and/or fluticasone propionate), plasma and tissue expression of both miRNAs significantly decreased and were no longer different from the controls. These microRNAs may serve as complementary non-invasive EoE markers and reduce the need for endoscopy/biopsies.

Autoimmune diseases (ADs) are a group of complex, chronic conditions characterized by disturbance of immune tolerance, with examples including systemic lupus erythematosus, rheumatoid arthritis, multiple sclerosis, and psoriasis. These diseases have unclear pathogenesis, and traditional therapeutic approaches remain limited. However, advances in high-throughput histology technology and scientific discoveries have led to the identification of various pathogenic factors contributing to ADs. Coupled with improvements in RNA nucleic acid-based drug synthesis, design, and delivery, RNA-based therapies have been extensively investigated for their potential in treating ADs. This paper reviews the progress in the use of miRNAs, lncRNAs, circRNAs, siRNAs, antisense oligonucleotides (ASOs), aptamers, mRNAs, and other RNA-based therapies in ADs, focusing on their therapeutic potential and application prospects, providing insights for future research and clinical treatment of autoimmune diseases.

Cardiac and kidney diseases are intimately linked through numerous pathophysiological pathways, frequently exerting reciprocal influences on one another. This interconnection often culminates in heightened morbidity and mortality rates within the clinical spectrum of cardiorenal syndrome (CRS). CRS is categorized into five types based on the primary organ involved and the chronicity of the condition. Each type of CRS encompasses a complex array of pathophysiological mechanisms. In recent years, the field of microRNAs (miRNAs) has risen to prominence, playing a crucial role in the pathogenesis of a multitude of diseases. By uncovering novel therapeutic targets through the study of miRNAs that influence the expression of the CRS genes, the prognostic outcomes for patients could be significantly improved. This article provides a comprehensive review, examining the pathophysiological underpinnings of CRS, miRNAs alterations and their associated mechanisms in various forms of CRS, as well as the potential of miRNAs in precision medicine and the use of miRNAs for the diagnosis of the disease.

Autophagy is a widespread physiological process in the body, which also protects the host by degrading invading pathogens and harmful substances during pathological conditions. Nevertheless, Mycobacterium tuberculosis (MTB), the causative agent of tuberculosis, has evolved strategies to subvert autophagy by modulating microRNA (miRNA) expression, enabling its escape from host defenses. In this study, we established an in vitro model using the human macrophage cell line infected with the highly virulent MTB strain H37Rv. Through RNA sequencing and bioinformatic analysis post H37Rv infection, we screened 14 differentially expressed miRNAs. We predicted and demonstrated that miR-30c-1-3p inhibits autophagy and promotes MTB survival by targeting ATG4B and ATG9B during the infection process. The results showed that miR-30c-1-3p expression was gradually increased before 12 h of H37Rv infection, followed by a decrease. Overexpression of miR-30c-1-3p suppressed autophagic activity. We also identified the targeting of miR-30c-1-3p to ATG4B and ATG9B for the first time, and overexpression of both ATG4B and ATG9B, alone or together, on the basis with upregulation of miR-30c-1-3p reversed the inhibition of autophagy. Autophagy levels were analyzed at different levels by western blot, immunofluorescence, and transmission electron microscopy, all of which showed that upregulation of miR-30c-1-3p inhibited autophagy during H37Rv infection. Additionally, the intervention of miR-30c-1-3p mimics resulted in an increased bacterial load in macrophages, suggesting that MTB achieves immune evasion by upregulating miR-30c-1-3p during infection. In conclusion, our study provides a valuable target for the development of host-directed anti-tuberculosis therapy as well as a new diagnostic marker.

Status epilepticus (SE) is a life-threatening disorder associated with neuronal pyroptosis. This study aims to explore the mechanism of HDAC1 in hippocampal neuronal pyroptosis induced by kainic acid in mice, providing a theoretical basis for SE treatment. A mouse model of SE was established by kainic acid. After sh-HDAC1 injection, the severity of SE and hippocampal neuronal damage were assessed. Cell model was established using kainic acid-induced HT22, followed by detection of HDAC1, miR-15a-5p, Caspase-1, cleaved Caspase-1, H3K9ac, and GSDMD-N using qRT-PCR and Western blot assays. Levels of IL-1β, IL-18, and LDH were measured. The enrichment of HDAC1 on the miR-15a-5p promoter was detected. The binding of miR-15a-5p to Caspase-1 was validated. We found that HDAC1 was highly expressed in kainic acid-induced SE. HDAC1 knockdown alleviated the symptoms of SE, inhibited cleaved Caspase-1, GSDMD-N, IL-1β, and IL-18, and suppressed hippocampal neuronal pyroptosis. HDAC1 bound to the miR-15a-5p promoter and reduced H3K9ac, thereby inhibiting miR-15a-5p expression. miR-15a-5p bound to Caspase-1 and inhibited Caspase-1 expression. Inhibiting miR-15a-5p or overexpressing Caspase-1 partially reversed the inhibitory effect of si-HDAC1 on kainic acid-induced cell pyroptosis. In conclusion, HDAC1 aggravates hippocampal neuronal pyroptosis in SE via the miR-15a-5p/Caspase-1 axis through deacetylation of H3K9.

Prostate cancer (PCa) is a frequently occurring malignant tumor affecting male reproductive system. miR-374a-5p was identified to participate in regulation of several tumors. The aim of the research was to discuss the influence for miR-374a-5p upon PCa progression and prognosis. A total of 112 PCa and 110 benign prostatic hyperplasia tissue samples were collected for the study. Real-time quantitative polymerase chain reaction was adopted to examine miR-374a-5p level in PCa tissues and cells. Kaplan-Meier and Cox model were applied to evaluate prognostic significance of miR-374a-5p for PCa. CCK8 and Transwell assays were carried out to analyze the efficacy of miR-374a-5p in PCa cell proliferation, migration and invasion. miR-374a-5p was under-expressed in PCa tissues and cells. Low expression of miR-374a-5p is linked to less favorable prognosis in PCa sufferers. Additionally, Cox analysis revealed that miR-374a-5p and TNM stage were two independent prognostic factors for PCa. Cellular assays showed that upregulating miR-374a-5p suppressed PCa cell proliferation, migration, and invasion.

Conversely, knockdown of miR-374a-5p facilitated PCa cell proliferation, migration, and invasion. miR-374a-5p expression decreased in PCa and was remarkably related to poor prognosis in PCa patients. miR-374a-5p acts in PCa by inhibiting cell proliferation, migration, and invasion. Consequently, miR-374a-5p has potential to act as a prognostic biomarker and a target for clinical therapeutic intervention in PCa.

Extracellular vesicles (EVs) are emerging as crucial biomarkers in molecular diagnostics, providing early detection of disease progression. Although ultracentrifugation remains the gold standard for vesicle isolation from biofluids, it has limitations in scalability and accessibility. This study presents lyophilization as an innovative method for preserving EVs and isolating microRNAs from saliva, utilizing its proven ability to maintain biological activity and prevent unwanted chemical reactions. We assessed five different sample preparation protocols combined with a dual-purification strategy. Structural and molecular integrity analyses revealed that lyophilized samples retained essential EV characteristics, including CD63/CD9 membrane localization. QELS analysis and electron microscopy confirmed distinct vesicle populations in both ultracentrifuged (30-50 nm and 320-360 nm) and lyophilized samples (50-70 nm and 360-380 nm). Importantly, lyophilized samples exhibited higher total RNA concentrations (p < 0.0001) while preserving key microRNA signatures (miR-16, miR-21, miR-33a, and miR-146b) with high fidelity. The efficacy of lyophilization is linked to its ability to systematically reduce solvent content through sublimation while maintaining vesicle integrity and molecular cargo. This method offers a practical, scalable alternative for EV isolation with significant implications for biomarker-based diagnostics.

Pancreatic ductal adenocarcinoma (PDAC) is the third leading cause of cancer-related death. Its poor prognosis is closely related to late-stage diagnosis, which results from both nonspecific symptoms and the absence of biomarkers for early diagnosis. MicroRNAs (miRNAs) exert a regulatory role in numerous biological processes and their aberrant expression has been found in a broad spectrum of diseases, including cancer. Cancer stem cells (CSCs) represent a driving force for PDAC initiation, progression, and metastatic spread. Our previous research highlighted the interesting behavior of miR-216a-5p and miR-125a-5p related to PDAC progression and the CSC phenotype. The present study aimed to evaluate the effect of miR-216a-5p and miR-125a-5p on the acquisition or suppression of pancreatic CSC traits. BxPC-3, AsPC-1 cell lines, and their CSC-like models were transfected with miR-216a-5p and miR-125a-5p mimics and inhibitors. Following transfection, we evaluated their impact on the expression of CSC surface markers (CD44/CD24/CxCR4), ALDH1 activity, pluripotency- and EMT-related gene expression, and clonogenic potential. Our results show that miR-216a-5p enhances the expression of CD44/CD24/CxCR4 while negatively affecting the activity of ALDH1 and the expression of EMT genes. MiR-216a-5p positively influenced the clonogenic property. MiR-125a-5p promoted the expression of CD44/CD24/CxCR4 while inhibiting ALDH1 activity. It enhanced the expression of Snail, Oct-4, and Sox-2, while the clonogenic potential appeared to be affected. Comprehensively, our results provide further knowledge on the role of miRNAs in pancreatic CSCs. Moreover, they corroborate our previous findings about miR-216a-5p's potential dual role and miR-125a-5p's promotive function in PDAC.

The objective of our study was to exploit the potential mechanism of microRNA-126-5p (miR-126-5p) in the occurrence and formation of allergic rhinitis (AR) in children.

Nasal mucosal tissues were obtained from AR in children and patients with adenoidectomy. Human nasal epithelial cell line (RPMI-2650) and BALB/c mice models were, respectively, established via ovalbumin (OVA) stimulation. Target genes and proteins levels were determined through quantitative real-time polymerase chain reaction (qRT-PCR) and enzyme-linked immunosorbent assay (ELISA) assays. The interaction of miR-126-5p with homeodomain-interacting protein kinase 2 (HIPK2) was confirmed via dual-luciferase reporter detection.

MiR-126-5p was memorably increased in nasal mucosal tissue specimens of AR children compared with patients with adenoidectomy, while HIPK2 was distinctly declined (all P<0.05). A negative association was found between miR-126-5p and HIPK2 expression (r=-0.5757, P<0.001). Moreover, HIPK2 was predicted to be targeted by miR-126-5p. Proinflammatory cytokines expressions were significantly increased, and anti-inflammatory cytokines were obviously decreased in AR RPMI-2650 cell model (P<0.001). NF-κB signaling pathway was also activated in AR RPMI-2650 cell model. MiR-126-5p inhibitor mitigated the stimulated function by OVA. Silencing HIPK2 recused miR-126-5p inhibitor phenomena in AR RPMI-2650 cell model. Furthermore, in vivo experiments further verified in vitro results, documenting that miR-126-5p inhibitor and si-HIPK2 relieved AR in the mice model.

MiR-126-5p down-regulation relieved inflammation response and events of AR in children and mice model of AR through HIPK2/NF-κB signaling pathway, suggesting being a latent therapeutic target in AR.