Endometriosis is a complex condition with a wide range of clinical manifestations, presenting significant challenges, particularly for young women. Its diverse and often perplexing presentations pose difficulties within the medical community. Laparoscopy remains the gold-standard diagnostic tool for endometriosis. However, alternative diagnostic methods are valuable for monitoring disease progression, assessing the likelihood of recurrence, reducing the need for surgical procedures, and facilitating timely decisions regarding fertility concerns. Recent research highlights the potential of microRNAs (miRNAs) as an alternative diagnostic test for endometriosis. A case-control study was conducted at the infertility unit of Arash Women's Hospital, involving 50 female participants, 25 with endometriosis and 25 without it. Plasma samples were collected and analyzed for the expression levels of 16 miRNAs using quantitative reverse transcription polymerase chain reaction (qRT-PCR). Diagnostic accuracy measures were evaluated to establish a reliable and comparable diagnostic framework. Compared to the control group, downregulation of 11 miRNAs and upregulation of 5 miRNAs were observed in the case group. Regarding expression patterns, evidence from this study indicates that half of the evaluated miRNAs fall into the high-agreement category with similar studies. Sensitivity (SN) of the evaluated miRNAs ranged from 64.0% to 88.0%, while specificity (SP) ranged from 56.0% to 88.0%. The area under the curve (AUC) was reported between 0.619 (miR-135a) and 0.846 (miR-340). These findings suggest that the evaluated miRNAs demonstrate moderate to acceptable diagnostic accuracy for endometriosis.

MicroRNAs (miRNAs) serve as important biomarkers for various diseases, including malignant tumors, and have broad applications in diagnosis, treatment, and prognosis. The development of real-time in situ imaging methods for monitoring miRNAs has both scientific and clinical value, and in this regard, catalytic hairpin assemblies (CHAs) can be used as precise and efficient nucleic acid circuits that facilitate hybridization without depleting targets. In this study, we developed a detection system based on CHA circuits encapsulated within CaCO3 nanoparticles, which represents a novel strategy for the detection of human pancreatic cancer. This encapsulation facilitates the pH-sensitive release of DNA probes, thereby ensuring the selective and sensitive detection of cancer-associated miRNAs. Our experimental results confirmed that the fabricated nanoparticles contributed to enhancing the stability and performance of the DNA circuits, thereby enabling precise miRNA detection and effective discrimination between cancerous and non-cancerous cells. Our findings in this study highlight the potential utility of CaCO3 nanoparticle-encapsulated CHA circuits for advancing miRNA-based cancer diagnostics and therapeutics.

Colorectal cancer is a major global health burden, with significant heterogeneity in clinical outcomes among patients. Identifying robust prognostic gene expression signatures can help stratify patients, guide treatment decisions, and improve clinical management. This review provides an overview of current prognostic gene expression profiles in colorectal cancer research. We have synthesized evidence from numerous published studies investigating the association between tumor gene expression patterns and patient survival outcomes. The reviewed literature reveals several promising gene signatures that have demonstrated the ability to predict disease-free survival and overall survival in CRC patients, independent of standard clinicopathological risk factors. These genes are crucial in fundamental biological processes, including cell cycle control, epithelial-mesenchymal transition, and immune regulation. The implementation of prognostic gene expression tests in clinical practice holds great potential for enabling more personalized management strategies for colorectal cancer.

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.

MicroRNAs (miRNAs) are non-coding RNAs that regulate gene expression. Among these, miR-660, located on chromosome Xp11.23, is increasingly studied for its role in cancer due to its abnormal expression in various biological contexts. It is regulated by 8 competing endogenous RNAs (ceRNAs), which adds complexity to its function. miR- 660 targets 19 genes involved in 6 pathways such as PI3K/AKT/mTOR, STAT3, Wnt/β-catenin, p53, NF‑κB, and RAS, influencing cell cycle, proliferation, apoptosis, and invasion/migration. It also plays a role in resistance to chemotherapies like cisplatin, gemcitabine, and sorafenib in lung adenocarcinoma (LUAD), pancreatic ductal adenocarcinoma (PDAC), and hepatocellular carcinoma (HCC), thus highlighting its clinical importance. Additionally, leveraging liposomes as nanocarriers presents a promising avenue for enhancing cancer drug delivery. Our comprehensive study not only elucidates the aberrant expression patterns, biological functions, and regulatory networks of miR-660 and its ceRNAs but also delves into the intricate signaling pathways implicated. We envisage that our findings will furnish a robust framework and serve as a seminal reference for future investigations of miR-660, fostering advancements in cancer research and potentially catalyzing breakthroughs in cancer diagnosis and treatment paradigms.

Cancer screening is an essential public health intervention for diagnosing cancers at an early stage that can enable earlier treatment - ideally with curative intent - and thus lead to improved outcomes. Over the past decade, liquid biopsy-based tests have emerged as a promising, minimally invasive and broadly applicable screening approach by combining multi-cancer early detection (MCED) with tumour tissue-of-origin identification. Large-scale randomized clinical trials evaluating liquid biopsy-based MCED approaches are now under way, although whether the diagnostic performance of this first generation of MCED tests is sufficient to translate into clinical benefits remains to be determined. In this Review, we discuss the promises and pitfalls of current MCED tests and highlight possible trajectories for the field of early cancer detection.

Overstretching of lung parenchyma may lead to injury, especially during mechanical ventilation. To date, there are no specific biomarkers of lung stretch, but transcriptomic signatures have not been explored. Our objective was to identify stretch-specific signatures using micro-RNA and gene expression.

Data on micro-RNA and RNA expression in response to stretch in experimental models were systematically pooled. Signatures were identified as those micro-RNAs or genes with differential expression in samples from stretched cells or tissues, and optimized using a greedy algorithm. Expression data was used to calculate transcriptomic scores. The accuracy of these scores was validated in animal models of lung injury, ex vivo mechanically ventilated human lungs, and bronchoalveolar lavage fluid (BALF, n = 7) and in serum samples (n = 31) of mechanically ventilated patients.

Six micro-RNAs (mir-383, mir-877, mir-130b; mir-146b, mir-181b, and mir-26b) were differentially expressed in stretched cell cultures (n = 24). Amongst the genes regulated by these micro-RNAs, a 451-gene signature was identified in vitro (n = 106) and refined using data from animal models (n = 143) to obtain a 6-gene signature (Lims1, Atp6v1c1, Dedd, Bclb7, Ppp1r2 and F3). Transcriptomic scores were significantly higher in samples submitted to stretch or injurious mechanical ventilation. The microRNA and RNA signatures were validated in human tissue, BALF and serum, with areas under the ROC curve between 0.7 and 1 to identify lung overdistention.

Lung cell stretch induces the expression of specific micro-RNA and genes. The potential of these signatures to identify lung stretch in a clinical setting must be explored.

Canine multicentric lymphoma is a common malignancy in dogs. It often responds well to initial chemotherapy but frequently relapses and has a poor response to subsequent treatment. B-cell (BCL) and T-cell (TCL) lymphomas differ in both their prognoses and chemotherapeutic treatment protocols. Currently, immunophenotyping can be costly and can only be performed on specific high-quality samples. MicroRNAs (miRNAs) are small molecules present in blood and tissues and are dysregulated in both human and canine lymphoma. We investigated 59 miRNAs by RT-qPCR to establish a serum miRNA profile in dogs with B-cell and T-cell multicentric lymphoma. Multiple miRNA pruned decision tree models were used to classify BCL and TCL cases from each other and controls, and to predict prognosis in BCL cases receiving standard CHOP chemotherapy. Six individual miRNAs were differentially expressed in serum between BCL and controls, and three were differentially expressed between BCL and TCL. A three-miRNA model (miR-155-5p, miR-1 and miR-181b) could differentiate between BCL, TCL and control samples with an accuracy of 83.02%. A three-miRNA model (miR-125b-5p, miR-350 and let-7b-5p) in BCL samples separated the cases into four groups with hazard ratios ranging from 0.44 to 3.5 for overall survival. This study established a serum miRNA profile for both BCL and TCL and demonstrated the utility of multiple serum miRNA models to assist in the diagnosis of lymphoma and BCL prognostication.

Cerebral cavernous malformation (CCM) is a hemorrhagic cerebrovascular disease where lesions develop in the setting of endothelial mutations of CCM genes, with many cases also harboring somatic PIK3CA gain of function (GOF) mutations. Rapamycin, an mTORC1 inhibitor, inhibited progression of murine CCM lesions driven by Ccm gene loss and Pik3ca GOF, but it remains unknown if rapamycin is beneficial in the absence of induction of Pik3ca GOF. We investigated the effect of rapamycin at three clinically relevant doses on lesion development in the Ccm3-/-PDGFb-icreERPositive murine model of familial CCM disease, without induction of Pik3ca GOF. Lesion burden, attrition, and acute and chronic hemorrhaging were compared between placebo and rapamycin-treated mice. Plasma miRNome was compared to identify potential biomarkers of rapamycin response. Outlier, exceptionally large CCM lesions (> 2 SD above the mean lesion burden) were exclusively observed in the placebo group. Rapamycin, across all dosages, may have prevented the emergence of large outlier lesions. Yet rapamycin also appeared to exacerbate mean lesion burden of surviving mice when outliers were excluded, increased attrition, and did not alter hemorrhage. miR-30c-2-3p, decreased in rapamycin-treated mouse plasma, has gene targets in PI3K/AKT and mTOR signaling. Progression of outlier lesions in a familial CCM model may have been halted by rapamycin treatment, at the potential expense of increased mean lesion burden and increased attrition. If confirmed, this can have implications for potential rapamycin treatment of familial CCM disease, where lesion development may not be driven by PIK3CA GOF. Further studies are necessary to determine specific pathways that mediate potential beneficial and detrimental effects of rapamycin treatment, and whether somatic PIK3CA mutations drive particularly aggressive lesions.

Liquid biopsy, a minimally invasive biopsy method that uses patient body fluids (e.g., blood, urine, or saliva), is considered a useful biomarker for early diagnosis, monitoring of tumor progression, and evaluating treatment efficacy. Extracellular vesicles (EVs), a diverse group of particles classified according to their size and biosynthetic method, are liquid bilayer structures released from various cells. EVs contain specific information, such as DNA, RNA, and proteins derived from released cells. Consequently, they have attracted attention for use in liquid biopsy. EV-derived microRNAs (miRNAs) and long noncoding RNAs (lncRNAs) are useful biomarkers for cancer diagnosis, tumor progression, and drug treatment resistance. Renal cell carcinoma (RCC), one of the most common type of urological cancer, accounts for 90% of all renal tumors. In contrast to prostate cancer, for which a tumor marker has been established, clinically applicable and useful biomarkers remain to be established for RCC. EV-derived miRNAs and lncRNAs have been identified as useful biomarkers in several types of carcinoma for determining the diagnosis and predicting tumor progression, and drug treatment resistance in patients with RCC. The development and identification of biomarkers to diagnose and predict tumor progression in RCC will improve the management and prognosis of patients with RCC. This review focuses on EV-derived miRNAs and lncRNAs and discusses the currently available EV-based biomarkers in RCC and their future prospects.

Small cell lung cancer (SCLC) is associated with high mortality and limited therapeutic options. There is increasing recognition that SCLC harbors molecular heterogeneity. Using a new liquid biopsy assay, it is demonstrated that SCLC subtypes, as determined by patient tumor tissue staining and cell lines, can be accurately identified by measuring the mRNA expression of subtype transcription factors (ASCL1, POU2F3, and NEUROD1) in circulating exosome-rich extracellular vesicles (Exo). Additionally, upregulation of Delta-like ligand 3 (DLL3) mRNA in Exo and its membrane protein (mProtein) in extracellular vesicles associated with tumor (tEV) may distinguish both limited- and extensive-stage SCLC patients from high-risk smokers, with AUC/ROC values of 0.836 and 0.839, respectively. By incorporating Exo-ASCL1 and Exo-POU2F3 mRNA expression with DLL3 Exo-mRNA/tEV-mProtein expression, the classifier enhances the AUC/ROC to 0.912 and 0.963 for limited- and extensive-stage SCLC patients, respectively.

MicroRNAs (miRNAs) are essential modulators of cardiac arrhythmias, influencing electrical remodeling, ion channel dynamics, and fibrosis. Dysregulated miRNAs, including miR-1, miR-21, and miR-328, contribute to arrhythmogenesis through altered ion channel expression, heightened fibrosis, and inflammation. Recent investigations have identified novel circulating miRNA signatures-such as hsa-miR-96-5p and hsa-miR-184 for post-operative atrial fibrillation (POAF) post-coronary artery bypass grafting, offering high predictive accuracy and targeting pathways like MAPK and TGF-β, as revealed by biological pathway analysis. These miRNAs serve as noninvasive biomarkers for early risk assessment. Therapeutically, miRNA inhibitors and mimics, enhanced by nanoparticle and viral vector delivery, provide targeted interventions. Despite challenges in specificity and delivery, miRNA-based approaches hold transformative potential for arrhythmia diagnosis and personalized management.

Heart failure (HF) is among the most important causes of worldwide morbidity, hospitalisation, and mortality. A reduction in anabolic hormonal axes seems to potentially play an important role in chronic HF progression and prognosis. Several lines of evidence support the critical roles of miRNAs in the endocrine system, and differentially expressed miRNA patterns were found to be able to detect HF. To date, the ability of miRNAs to detect HF patients affected by hormonal deficiencies has yet to be addressed. The aim of this study was to explore the association between circulating miRNA profiles and multiple hormonal deficiencies in HF patients to provide new insights into HF pathophysiology. The study cohort included 129 subjects (94 HF patients and 35 controls). Circulating miRNAs assayed in plasma samples were miR-1-3p, miR-10b-5p, miR-24-3p, miR-193a-5p, miR-454-3p, miR-503-5p, miR-551b-3p, and miR-598-3p. NT-proBNP, IGF-1, fT3, DHEA-S, testosterone, HF subtypes, and NYHA class were also evaluated. A multiple hormonal deficiency syndrome (MHDS) was defined as the presence of ≥two hormone deficiencies. We found that miR-10b-5p, miR-193a-5p, and miR-1-3p could distinguish chronic HF patients from controls. The identified miRNAs were downregulated in HF patients, particularly those with NYHA I-II classifications and pathological values of NT-proBNP. In addition, these three circulating miRNAs correlated with each other, and their deregulation seems to be influenced by hormone deficiencies, especially in patients with reduced ejection fraction. Among the three miRNAs, miR-10b-5p was the best able to diagnose chronic HF-MHDS patients (AUC = 0.8). These results support the clinical utility of miR-10b-5p, miR-193a-5p, and miR-1-3p in detecting HF patients, especially those with hormone deficiencies.

Multiple endocrine neoplasia type 1 (MEN1) is a rare hereditary tumor syndrome caused by inactivating mutations of the MEN1 gene and characterized by the occurrence of multiple endocrine tumors within a single patient (i.e., parathyroid, pituitary, and pancreatic neuroendocrine tumors (NETs)). However, the lack of a genotype-phenotype correlation does not allow individual disease evolution to be foreseen. Epigenetic factors, such as microRNAs, are suspected to contribute to MEN1 tumorigenesis, presumably explaining the lack of genotype-phenotype association. Our previous studies indicated miR-24-3p, miR-1301-3p, miR-664a-3p, and miR-4258 as potentially involved in MEN1 parathyroid tumorigenesis. In this study, we examined the expression of two circulating microRNAs (c-miRNAs), miR-24-3p and miR-1301-3p, in the serum of MEN1 patients. c-miRNAs were evaluated by RT-qPCR in serum collected from 25 MEN1 patients and 25 age- and gender-matched healthy volunteers (HCs). Receiver operating characteristic (ROC) curves were constructed to determine miRNA sensitivity and specificity. RT-PCR analysis revealed that expression levels of circulating miR-1301-3p were significantly downregulated, while those of miR-24-3p were significantly upregulated in the serum of MEN1 patients compared to HCs. Additionally, ROC analysis exhibited a good diagnostic power for both miRNAs (area under the ROC curve (AUC) values: 0.7356 and 0.7928 for miR-1301-3p and miR-24-3p, respectively) in distinguishing MEN1 patients from matched HCs. These preliminary data suggest circulating miR-1301-3p and miR-24-3p as potential non-invasive diagnostic biomarkers for MEN1 syndrome, regardless of different clinical phenotypes and MEN1 mutation types.

Rolling circle amplification (RCA) at ambient temperature is prone to false positive signals during nucleic acid detection, which makes it challenging to establish an efficient RCA detection method. The false positive signals are primarily caused by binding of non-target nucleic acids to the circular single-stranded template, leading to non-specific amplification. Here, we present an RCA method for miRNA detection at 37 °C using two circular ssDNAs, each of which is formed by ligating the intramolecularly formed nick (without any splint) in a secondary structure. The specific target recognition is realized by utilizing low concentrations (0.1 nM) of circular ssDNA1 (C1). A phosphorothioate modification is present at G*AATTC on C1 to generate a nick for primer extension during the primer self-generated rolling circle amplification (PG-RCA). The fragmented amplification products are used as primers for the following RCA that serves as signal amplification using circular ssDNA2 (C2). Notably, the absence of splints and the low concentration of C1 significantly inhibits non-target binding, thus minimizing false positive signals. A high concentration (10 nM) of C2 is used to carry out linear rolling circle amplification (LRCA), which is highly specific. This strategy demonstrates a good linear response to 0.01-100 pM of miRNA with a detection limit of 7.76 fM (miR-155). Moreover, it can distinguish single-nucleotide mismatch in the target miRNA, enabling the rapid one-pot detection of miRNA at 37 °C. Accordingly, this method performs with high specificity and sensitivity. This approach is suitable for clinical serum sample analysis and offers a strategy for developing specific biosensors and diagnostic tools.

MicroRNAs (miRNAs) are small non-coding RNA molecules that function as gene regulators in physiological processes, including proliferation, differentiation, and apoptosis. Various microRNAs have been linked to pathophysiological events associated with heart disease. In this case-control study, we investigated the levels of human miR-21, miR-23a, miR-142-5p, and miR-126 among heart failure patients with reduced ejection fraction (HFrEF), compared to healthy control participants.

We prospectively enrolled clinically stable patients with heart failure (HF) and left ventricle ejection fraction (LVEF) ≤ 40%, and healthy individuals. MicroRNAs were analyzed from venous blood, using a microRNA enzymatic immunoassay (miREIA) method. Plasma miRNA levels were compared between HFrEF patients and healthy individuals, using non-parametric tests.

We enrolled 73 patients with HFrEF (86% males, mean age: 66.3 ± 10.7 years) and 99 healthy subjects (36% males, mean age: 44.7 ± 15.9 years). All four assayed miRNAs exhibited significantly higher median levels in heart failure patients compared to healthy controls: miR-21p, 243 pmol·l-1 vs. 14 pmol·l-1; miR-23a-3p, 705 pmol·l-1 vs. 119 pmol·l-1; miR-142-5p, 1,695 pmol·l-1 vs. 146 pmol·l-1; and miR-126-3p, 528 pmol·l-1 vs.21 pmol·l-1 (P ≤ 0.001 for all). The analyzed miRNA levels did not differ according to age, weight, height, or body mass index. No miRNA levels correlated with NTproBNP levels.

Our findings revealed that the levels of miR-21-5p, miR-23a-3p, miR-142-5p,and miR-126-3p were significantly higher among HFrEF patients compared to healthy controls. Further exploration of these miRNAs may lead to new diagnostic, prognostic, and therapeutic options for HF patients.

Intervertebral disc degeneration (IDD) is a primary cause of degenerative disc disease; however, the mechanisms underlying it remain unknown. Although great efforts have been made to develop new regenerative therapies, their clinical success is limited. Recent research has indicated that microRNAs (miRNAs) are significantly involved in the progression of IDD. Investigating the role of miRNA intervention in IDD could facilitate the development of therapeutic strategies based on miRNAs. However, circulating miRNAs have not yet been recognized as standard biomarkers for IDD. In this study, we observed that the expression of miR-27b-3p was elevated in the blood and nucleus pulposus (NP) tissue of patients with IDD. Furthermore, reducing the expression of miR-27b-3p was shown to impede the progression of IDD. MiR-27b-3p could reduce the expression of collagen II and ACAN and promote the expression of MMP13 and ADAMT-5 in vitro and in vivo. miR-27b-3p aggravated IDD progression by directly targeting peroxisome proliferator-activated receptor gamma (PPARG), a negative regulator of the NF-κB signal pathway. This study also established that PPARG serves a protective role in IDD. The overexpression of PPARG was able to mitigate the detrimental effects caused by miR-27b-3p in NP cells and animal models of IDD, indicating that miR-27b-3p facilitates the progression of IDD through its interaction with PPARG. Additionally, the transcription factor ATF1 was found to enhance the expression of miR-27b-3p by targeting its promoter region, thereby promoting the degenerative impact of miR-27b-3p on NP cells. Given that miR-27b-3p can promote IDD both in vitro and in vivo, it holds potential as a biomarker, and the inhibition of miR-27b-3p expression may represent a novel therapeutic target for IDD.

MicroRNAs have emerged as potential biomarkers of liver injury during organ transplantation due to their specificity, easy detection and stability in many biofluids. Heparin, which has a well-known inhibitory effect on RT-qPCR based measurements, is commonly used during organ donation. Heparinase I treatment has been used to overcome the inhibiting effect of heparin in RNA RT-qPCR analysis. However, there is a lack of evidence regarding its effective, feasible use improving specific miRNA quantification yield in the liver transplant setting. The aim of this study is to evaluate the effect of heparinase I on miRNA detection levels by RT-qPCR in different samples from liver donors.

Prospective, single-centre study including evaluation of liver biopsy, perfusate fluid and serum from deceased organ donors from October 2019 to May 2021. Samples from brain death donors (DBD, n = 4) and donors after circulatory death recovered with abdominal normothermic regional perfusion (DCD n = 4) were analysed for the presence of liver-injury related miRNAs (miR-122 and miR-148a) in the absence or presence of heparinase I (6 IU or 12 IU) to evaluate its effect on miRNA detection levels by RT-qPCR. A subgroup of heparinized serum samples from patients undergoing cardiopulmonary bypass was analysed for validation purposes. The study is registered with ClinicalTrials.gov (NCT06611046), and accrual is complete.

The expression of miR-122 relative to reference genes was 44.5, 16.8 and 4.2-fold higher in liver biopsy, perfusates and serum respectively, while miR-148a was 3.4, 2.2 and 2.6-fold higher, without differential expression between donor groups (p > 0.05). Heparinase I treatment did not improve PCR results and affected miRNA detection yields in a dose-dependent way with delayed and dispersed Ct values. In highly heparinized DCD serum samples, heparinase I treatment significantly reduced the relative expression of miR-122 and miR-148a compared to non-treated samples, 2-fold and 6.1-fold, p < 0.05 respectively. Moreover, treatment with heparinase I led to a rise in lost values, from 12.5% to 25% in perfusates and 67.7% to 68.7% in serum samples treated with 6IU and 12IU of heparinase I respectively.

The need for heparinase I treatment to overcome RNA quantification interference in heparinized samples should be addressed in each individual analysis. Heparin inhibition seems variable among miRNAs, and the additional handling with heparinase may affect reliable miRNA quantification due to RNA degradation, introducing bias in gene expression interpretation.

MicroRNAs (miRNAs) are emerging as circulating biomarkers in germ cell tumors (GCT) with potential to guide management. Their role and expression patterns are more established in testicular GCTs, while lesser data exist in ovarian GCTs (OGCT).

Patients diagnosed with OGCT with plasma and tumor tissue available in our provincial biobank were included. Total RNA was extracted, and RT-qPCR was performed to measure miR-371-3 and miR-302/367 levels. Healthy plasma and ovarian tissue served as controls. Statistical analyses were performed using ANOVA and the Mann-Whitney U test. Clinicopathologic data was collected by chart review.

From 2007 to 2022, 23 patients with OGCT were identified: 13 with viable non-teratoma germ cell (VNTGC) and 10 with immature teratoma germ cell (ITGC) tumors. Compared to healthy controls, all patients with VNTGC but not ITGC tumors had significantly higher miRNA levels in preoperative plasma and tumor tissue. Plasma miRNA kinetics correlated with disease burden, decreasing to undetectable levels following treatment, and increasing significantly upon relapse.

MiR-371-3 and miR-302/367 are highly expressed in ovarian VNTGC but not ITGC tumors, and their plasma levels correlate with disease burden. Future studies validating these findings in a larger cohort are needed to develop miRNAs as circulating biomarkers for clinical use.

Silver nanoclusters (Ag NCs) are widely applied in the biosensing of metal ions, small organic molecules, nucleic acids, amino acids, and proteins due to their particular fluorescence and chemical properties. Organic matrices such as DNA are usually employed for Ag NC synthesis and stabilization. They make Ag NC/matrix complexes biocompatible and sensitive to the environment. It has recently been shown that Ag NCs based on DNA matrices are capable of self-assembly and rearrangement followed by a change in the fluorescence and absorbance characteristics. These attributes allow the development of sensors with target molecule detection visible even by the naked eye. Here we suggest a simple one-step turn-on highly specific microRNA-210 sensor based on a fluorescent Ag NC. The main feature of the sensor is the smart design of a binary matrix, which provides the appearance of a bright green fluorescence signal only after Ag NCs/DNA-matrix complexes are bonded to the target sequence. The microRNA detection assay requires no additional action because the process proceeds by itself. A comprehensive optimization of the binary probe structure and location was carried out. An approach to detection leading to minimal background signal was defined as follows. The approach involves the preliminary synthesis of non-fluorescent silver clusters using a single strand of the binary matrix containing a 5'-CCCGTTTT-3' part. It was shown that these "dark" structures can be stored for at least a month before analysis. The fluorescence intensity of the green Ag NCs increases in the presence of the microRNA-210 sequence, and that dependence on the target concentration tends to be linear in the range of 5-500 nM. The sensor demonstrates specificity to the miR-210 sequence, and the LOD (limit of detection) was established as 5 nM in serum samples.

Gastric cancer (GC)，the fourth leading cause of cancer-related deaths globally and thus early detection, is considered critical for diagnosis and treatment of this disease. It is well known that measurement of microRNA (miRNA) may serve as diagnostic and prognostic biomarker for GC. The aim of this study was to determine whether miR-30c was present in patients with gastric cancer and to correlate relative expression with patient survival. A total of 162 GC patients and 150 healthy controls were recruited. miR-30c levels were quantified in serum using quantitative real-time PCR(QRT-PCR). The sensitivity and specificity of circulating miR-30c was compared to carbohydrate antigen (CA) CA72-4, CA19-9, and carcinoembryonic antigen (CEA), 3 known markers associated with GC. QRT-PCR demonstrated downregulation of gene expression of miR-30c in GC patients. Downregulation of miR-30c gene expression was significantly correlated with stage of cancer, lymphatic metastasis, and distal metastasis. The sensitivity to detect GC of miR-30c, CA72-4, CA19-9, and CEA in serum of GC was 80%, 43%, 21%, and 42%, respectively, while specificity was 89%, 57%, 30%, and 78% respectively. Kaplan-Meier survival analysis showed that the presence of low gene expression of miR-30c was effective in predicting poor prognosis in GC patients. Our data suggest that circulating serum miR-30c concentrations may serve as a reliable biomarker for GC occurrence. (212words).

Nephrotic syndrome is a complex renal condition characterized by abnormal protein permeability into the urine space, leading to edema and renal failure. Recent research suggests that deregulation of microRNAs contributes to the pathogenesis of this disease. MicroRNAs are small, non-coding RNA molecules that regulate gene expression by binding to complementary messenger RNA sequences. In this study, we employed bioinformatics techniques to analyze microRNA expression in urine samples from nephrotic syndrome patients and healthy control participants. Our results revealed a significant disruption of microRNA expression profiles in patients with nephrotic syndrome, indicating that these microRNAs may play a crucial role in the disease. This study highlights the potential of urinary microRNAs as biomarkers for nephrotic syndrome and warrants further investigation into their functional significance in the disease pathogenesis.

To research the diagnostic value and mechanism of miR-1281 involved in type 2 diabetes mellitus (T2DM) and diabetic kidney disease (DKD).

One hundred and eighteen patients with T2DM (68 DKD patients) and 35 healthy individuals were included. Fasting venous blood and one-time morning urine were collected for biochemical testing. RT-qPCR detected miR-1281 expression, ROC curve assessed the diagnostic value of miR-1281 for T2DM and DKD, and logistic regression predicted risk factors affecting the progression of DKD. ELISA analysed inflammatory cytokine expression, and Pearson correlation assessed its relevance to miR-1281. CCK8 detected cell proliferation and flow cytometry recorded apoptosis.

miR-1281 was upregulated in T2DM patients and increased more significantly in DKD patients. The ROC curves indicated that miR-1281 had diagnostic value in predicting T2DM and DKD, and miR-1281 was closely related to the pathological characteristics of DKD patients. The logistic results showed that increased miR-1281 expression was a risk factor for DKD progression. ELISA showed that inflammatory cytokines (IL-6, IL-18, TNF-α) were significantly increased in patients with T2DM, and Pearson correlation analysis indicated a positive correlation between miR-1281 and inflammatory cytokine expression. The high-glucose environment promoted cell proliferation, decreased apoptosis, and increased inflammatory factor levels, but transfection of the miR-1281 inhibitor resisted the adverse effects of the high-glucose environment on cells.

miR-1281 promotes glomerular cell proliferation, inhibits apoptosis, and increases the level of intracellular inflammation, leading to impaired renal function in T2DM patients.

MicroRNAs (miRNAs) serve as potential biomarkers for many diseases such as cancer, neurodegenerative diseases and cardiovascular conditions. The portable and accurate detection of miRNA is of great significance for the early diagnosis, treatment optimization and prognostic evaluation of diseases. Herein, a photothermal/visual dual-mode assay for let-7a is developed utilizing oxidized 3, 3', 5, 5' - tetramethylbenzidine (oxTMB) as signal reporter. Let-7a efficiently initiates two cascade amplification processes of catalytic hairpin assembly (CHA) and hybridization chain reaction (HCR) on the magnetic microspheres. It is worth noting that the hairpin DNA employed for HCR is labeled with horseradish peroxidase (HRP). Consequently, a substantial accumulation of HRP occurs on the magnetic microspheres and further catalyzes the oxidation of TMB by H2O2 to oxTMB. The blue color of oxTMB enables its detection through visual sensing, while the photothermal conversion characteristic of oxTMB allows for its detection through photothermal sensing. The concentration of miRNA is positively correlated with the enrichment of HRP on magnetic microspheres, which in turn affects the production of oxTMB. Therefore, the photothermal and visual dual-mode assay for let-7a can be indirectly realized by the detection of oxTMB. The dual signals are cross-validated to effectively reduce false positives. Precise miRNA quantification is achieved using a household thermometer, which is conducive to the popularization of this proposed strategy. Moreover, by adjusting the hairpin DNA sequences utilized for CHA and HCR, the dual-mode assay platform has a potential to serve as a universal approach for other biomarkers.

In recent years, iodine deficiency-related diseases have been effectively controlled; the prevalence of excessive iodine-induced thyroid diseases has increased, such as hyperthyroidism. However, there are still several controversial outcomes regarding the relationship between excessive iodine intakes and hyperthyroidism. MicroRNAs (miRNAs) extensively participate in the progression of thyroid diseases; nevertheless, the relationship and mechanism between iodine exposure and miRNAs have not been explored in hyperthyroidism patients. In this study, a total of 308 pairs of hyperthyroidism patients and healthy controls were enrolled in. Logistic regression analysis showed that level of water iodine >100 μg/L was an independent risk factor for hyperthyroidism. Compared with the healthy control, the serum thyroglobulin (Tg) content and levels of interferon-γ (IFN-γ), interleukin 6 (IL-6), and tumor necrosis factor-α (TNF-α) were significantly elevated in hyperthyroidism patients. Further, high-throughput miRNA sequencing was applied to find crucial miRNAs involved in the occurrence of hyperthyroidism related to excessive water iodine. Based on the fold change and Q value, miR-144-3p, miR-204-5p, miR-346, miR-23b-5p, and miR-193b-3p were selected for validation by qRT-PCR. Our results showed that miR-346 and miR-204-5p in the case group were significantly lower than those of the control group, and the similar results found under the level of water iodine >300 μg/L. Nonetheless, no significant difference was found at 10-100 μg/L level of water iodine. Furthermore, the ROC curve indicated that miR-346 and miR-204-5p had the ability to diagnose hyperthyroidism patients. Taken together, excessive water iodine may decrease the expression of miR-346 and miR-204-5p, which mediate the elevation of Tg and cytokines, ultimately making contribution to the development of hyperthyroidism.

Type 2 diabetes mellitus (T2DM) is a medical disorder characterized by high blood sugar levels resulting from a lack of insulin caused by impaired activity of 𝛽-cells and/or the inability of insulin to efficiently transport glucose from the bloodstream into cells, a condition referred to as insulin resistance. This occurs not only in insulin-sensitive tissues such as muscles, adipose tissue, and the liver, but also in the gastrointestinal tract, which may be caused by a defect in the insulin signaling pathway. MicroRNAs (miRNAs) are RNA molecules that do not code for proteins and play a role in multiple pathways. Several studies have suggested that specific miRNAs could potentially be used as biomarkers for diagnosing diabetes. These miRNAs regulate the formation of pancreatic islets, the differentiation of β-cells, the secretion of insulin, and the control of glucose metabolism. miRNA-mediated pathways are associated with human genetic illnesses resulting from mutations in the maturation process of miRNAs. The changes in miRNAs impact their ability to bind to mRNA targets, hence modifying gene expression. This review provides a concise overview of the latest studies investigating the correlation between miRNA expression and the regulation of glucose levels in cases of β-cell malfunction and insulin resistance.

Dysregulation of microRNA (miRNA) expression in the brain is a common feature of neurodegenerative diseases. Beyond their conventional role in regulating gene expression at the post-transcriptional level, certain miRNAs can act extracellularly as signaling molecules. Our study elucidates the identity of such miRNA species serving as ligands for membrane receptors expressed in central nervous system (CNS) neurons and the impact of such miRNAs on neurons in the context of neurodegenerative disease.

We combined a machine learning approach with the analysis of disease-associated miRNA databases to predict Alzheimer's disease (AD)-associated miRNAs as potential signaling molecules for single-stranded RNA-sensing Toll-like receptors (TLRs) 7 and 8. TLR-expressing HEK-Blue reporter cells, primary murine microglia, and human THP-1 macrophages were used to validate the AD miRNAs as ligands for human and mouse TLR7 and/or TLR8. Interaction between mouse cortical neurons and extracellularly applied AD miRNAs was analyzed by live cell imaging and confocal microscopy. Transcriptome changes in cortical neurons exposed to AD miRNAs were assessed by RNAseq and RT-qPCR. The extracellular AD miRNAs' effects on CNS neuron structure were investigated in cell cultures of murine primary cortical neurons and iPSC-derived human cortical neurons by immunocytochemistry. We employed a mouse model of intrathecal injection to assess effects of AD miRNAs acting as signaling molecules on neurons in vivo.

We identified the AD-associated miRNAs miR-124-5p, miR-92a-1-5p, miR-9-5p, and miR-501-3p as novel endogenous ligands for TLR7 and/or TLR8. These miRNAs being extracellularly stable and active were taken up by murine cortical neurons via endocytosis and induced changes in neuronal inflammation-, proliferation-, and apoptosis-related gene expression. Exposure of both murine and human cortical neurons to the AD-associated miRNAs led to alterations of dendrite and axon structure, synapse protein expression, and cell viability in a sequence-dependent fashion. Extracellular introduction of the AD miRNAs into the cerebrospinal fluid of mice resulted in both changes in neuronal structure and synapses, and neuronal loss in the cerebral cortex. Most of the observed extracellular miRNA-induced effects on cortical neurons involved TLR7/8 signaling.

Neurodegenerative disease-associated miRNAs in extracellular form act as signaling molecules for CNS neurons including human cortical neurons, thereby modulating their structure and viability.

While mesangial IgA deposition is the pathognomonic feature of IgA nephropathy (IgAN), the extent of mesangial IgA accumulation does not correlate with the future risk of kidney failure. This has led to the search for other serum factors that may influence clinical outcome. The emergence of microRNAs (miRs) as negative regulators of gene expression and the increasingly recognized role of extracellular miRs in intercellular communication has prompted study of the influence of miRs on inflammatory and scarring pathways in the kidneys.

Here, next generation sequencing and subsequent qPCR validation identified a significant increase in the serum levels of miR-483-5p, largely packaged within exosomes.

Levels of miR-483-5p in serum exosomes were greatest in those IgAN patients with higher levels of proteinuria who subsequently developed kidney failure. Exosomal miR-483-5p content significantly correlated with numerous soluble isoforms of the tumor necrosis factor (TNF) receptor super family suggesting lymphocytes as a source of the miR-enriched exosomes. In PBMC miR-483-5p expression was almost exclusively seen in CD19+ lymphocytes. Activation of a human IgA secreting B-cell line with soluble TNFR1 induced miR-483-5p synthesis and enrichment within exosomes. Exposure to miR-483-5p-enriched B cell exosomes resulted in a proinflammatory phenotypic change in cultured human collecting duct epithelial cells, likely mediated through suppression of the transcription factor SOCS3. miR-483-5p-enriched exosomes were also present in the urine of patients with IgAN.

Interaction of B lymphocyte-derived miR-enriched exosomes with tubular epithelial cells may provide an explanation for the progressive tubulointerstitial scarring and loss of kidney function seen in IgAN.

MicroRNAs (miRNAs) are essential regulators of gene expression and are significantly involved in both preventing and treating a range of diseases. To that end, we developed an ultra-sensitive detection method for miRNA-141 by integrating exonuclease-assisted target recycling signal amplification with the CRISPR/Cas12a system. This method employs a variable hairpin probe (HP) designed to hybridize with miRNA, which, under the action of exonuclease III (ExoIII), cleaves the hairpin probe and triggers target recycling signal amplification. This results in the formation of output DNAs (ODs) containing multiple repeat sequences. The CRISPR/Cas12a system identifies these repeated sequences in ODs through its crRNA component, which in turn triggers the trans-cleavage function of the Cas12a/crRNA complex. It leads to the cleavage of a fluorescently quenched reporter probe. Consequently, this process restores fluorescence, producing a significantly enhanced fluorescent signal that facilitates the detection of miRNA-141, achieving a detection threshold down to 62 fM. This detection approach can specifically differentiate miRNA-141 from other confounding substances and has effectively identified low concentrations of miRNA-141 in actual sample human serum and diverse cancer cell lysates, showcasing its capability for tracing various nucleic acid biomarkers at minimal levels.

Since microRNAs are released into the bloodstream and miRNA profiles are supposed to differ between healthy and tumour patients, miRNAs seem to have potential as biomarkers. An essential prerequisite for biomarkers in a routine diagnostic setup is their stability in serum over time. In this study, serum samples from 10 healthy dogs were analysed at different time points and under various temperature conditions (after 24 and 48 h, at 4° or 20 °C) for the copy number of eight miRNAs (miR-20b, 21, 122, 126, 192, 214, 222, 494) using ddPCR. The miR-21 had the highest copy number, whereas miR-494 had the lowest copy number in canine blood samples. The values of each miRNA varied individually between the dogs, showing a 5 to 10-fold range. Stability differed between the miRNAs, with miR-192 having the best stability. The copy number of miR-20b, miR-126 and miR-214 decreased not significantly during 48 h storage time. In contrast, miR-21, miR-122 and miR-222 were stable for 24 h only but decreased significantly after 48 h. The (in)stability of individual canine miRNAs must be considered when transferring study results into veterinary routine diagnostics, as the transport and storage conditions are variable. As far as possible, standardisation of sampling, storage and quantification of miRNAs is needed.

The 2024 Nobel Prize in Physiology or Medicine was awarded to the pioneers who reported that microRNAs (miRNAs) regulate and direct the switch between physiological and pathological pathways via their over- or underexpression. The discovery changed the medical landscape and there are many completed and on-going clinical studies based on miRNAs. MiRNAs occur at the femtomolar level in biological fluids and are typically quantified using amplification-based techniques. Experimental nanopores have illustrated potential for trace analysis including amplification-free miRNA quantification. We repurposed the MinION, the only commercially available nanopore array device, and developed unique probes and protocols to detect and measure miRNA copies in blood and urine. Here, we report that miRNA copies are proportional to the total RNA isolated from the biospecimen, and that three known miRNA cancer biomarkers, i.e., miR-21, miR-375, and miR-141, were more than 1.5-fold overexpressed in blood samples from breast, ovarian, prostate, pancreatic, lung, and colorectal cancer patients compared to healthy patients. In these cancer samples, miR-15b was not overexpressed, in agreement with earlier studies. In contrast to literature reports, sample variability was undetectable in this study. The potential and limitations of this ready-to-use MinION/Yenos platform for multiple-cancer early detection (MCED) using blood or urine are discussed.

Mucosal healing in Crohn's disease (CD) has been established as a crucial target of treatment, leading to long term remission and decrease in complication rates. Endoscopy still serves as the gold standard for assessment, particularly in the small bowel where balloon or capsule enteroscopy is frequently needed. However, these modalities are often unavailable, expensive, and invasive, posing risks to patients. Consequently, the identification of accessible and reliable biomarkers, especially in small intestinal CD, remains a challenge. The study by Ohno et al, published in this issue, further illuminates this field. It confirms the potential role of fecal biomarker leucine-rich α2 glycoprotein (LRG) and validates findings from previous smaller trials. Comparing to other markers LRG showed a much higher predictive value for mucosal healing of the small bowel, making it a useful option for small intestinal CD follow up. In this editorial, we explore the optimal marker of inflammation or mucosal healing in CD, particularly in the small bowel. We provide an overview of available conventional biomarkers and introduce several novel biomarkers, including an update on emerging technologies and innovations.

Aberrant microRNA expression is associated with tumor progression in various organs. Detecting microRNAs as clinical cancer biomarkers can facilitate early cancer diagnosis and monitoring. However, the rapid and accurate quantification of microRNAs from biological samples remains a significant challenge. Here we developed a one-pot isothermal assay utilizing a molecular circuit with CRISPR/Cas13a (CRISPR-circuit) to rapidly convert, amplify and report different microRNAs within 15 min at the attomolar (aM) level. Then the full process was performed on an active centrifugal microfluidic chip and its corresponding portable equipment for parallel detection of multiple microRNAs, including miR-21, miR-141, miR-196a, and miR-1246. We also demonstrated its application for identifying cell lines and clinical samples of cancer patients with varying microRNA levels, which showed a strong correlation with the RT-qPCR. The assay can be easily adapted for the detection of any microRNA by simply modifying the converter primer, thereby holding significant potential for accurate disease detection and clinical diagnosis.

Prostate cancer (PCa) is a highly life-threatening disease in men, causing numerous deaths worldwide. As PCa is often diagnosed at a late stage, current diagnostic methods can be invasive and sometimes lead to unnecessary treatments. Therefore, new non-invasive approaches are needed to detect biomarkers for more rapid and accurate PCa diagnosis. Exosomes, extracellular vesicles, provide valuable insights into cellular health and disease progression. Recent studies have indicated the potential use of exosomes as biomarkers for diagnosing PCa. Developing fast, reliable, and sensitive methods for exosome detection is essential. Biosensors, powerful analytical tools for biological samples, have become increasingly crucial in exosome analysis. This review summarizes recent advancements in biosensor technology for exosome detection and provides insights into future perspectives. The goal is to encourage innovative biosensor-based approaches for exosome detection and contribute to the early diagnosis and clinical monitoring of various diseases.

Pancreatic cancer is recognized as one of the most lethal types of cancer globally, characterized by a high mortality rate and a bleak prognosis, which greatly contributes to cancer-related deaths. Forecasts suggest that by 2030, pancreatic cancer will exceed other cancer types in prevalence. The disease presents considerable difficulties owing to the lack of prominent symptoms in its early stages, restricted options for early detection, rapid progression, and unfavorable outcomes. Presently, traditional methods for diagnosing pancreatic cancer primarily rely on imaging techniques. However, these methods often entail significant costs, require considerable time, and necessitate specialized skills for both operating the equipment and interpreting the resulting images. To overcome these obstacles, the use of biosensors has been proposed as a potentially valuable tool for the early detection of pancreatic cancer. MicroRNAs (miRs), a type of small non-coding RNA molecules, have emerged as highly sensitive molecular diagnostic tools that have the potential to function as precise indicators for a range of diseases, including cancer. Biosensors have been suggested as a potential solution for tackling these challenges, offering a promising approach for the early detection of pancreatic cancer. Small non-coding RNA molecules known as MicroRNAs (miRs) have become recognized as extremely sensitive molecular diagnostic tools and can act as precise biomarkers for different diseases, such as cancer. Moreover, this manuscript presents a thorough summary of the latest innovations in nano-biosensors that have been specifically developed for the identification of non-coding RNAs related to pancreatic cancer.