Serum miR-146a Research Articles

Significance of serum miR-29a in the occurrence and progression of diabetic nephropathy: A cross-sectional study.

BackgroundDiabetic nephropathy (DN), a common microvascular complication of type 2 diabetes mellitus (T2DM), is an important factor causing chronic kidney disease. However, the relationship between miR‐29a and DN remains unknown. Therefore, a cross‐sectional study was conducted to identify a potential molecular biomarker for DN prevention and management by detecting the serum miR‐29a levels.MethodsThe serum miR‐29a levels were measured in 360 subjects (180 T2DM patients and 180 healthy controls) using quantitative reverse transcription PCR (qRT‐PCR), and other conventional indicators were measured and analysed. A binary logistic regression was used to evaluate the DN risk factors; a receiver operating characteristic (ROC) curve was applied to analyse the diagnostic efficacy of miR‐29a for DN, and a Spearman's rank correlation analysis was used to evaluate the correlation between serum miR‐29a and cystatin C.ResultsThe serum miR‐29 levels in the T2DM patients were higher than those in the healthy subjects and significantly increased with the progression of DN (p < 0.05). Serum miR‐29a and cystatin C are independent predictors of the occurrence of DN. Compared with a single indicator, the combination of serum miR‐29a and cystatin C has better DN diagnostic performance. In addition, the serum miR‐29a levels were positively correlated with cystatin C in the patients with DN (r = 0.521, p < 0.001).ConclusionThe expression of serum miR‐29a was significantly associated with the occurrence and progression of DN and is expected to become a potential biomarker for the diagnosis of DN.

MiR-181a Ameliorates the Progression of Myasthenia Gravis by Regulating TRIM9.

Abnormally activated CD4+ T cells are considered to be an important factor in the pathogenesis of myasthenia gravis (MG). In the pathogenesis of MG, the imbalance of proinflammatory cytokines and immune cells maintains the imbalance of immune response and inflammatory microenvironment. Studies have shown that miRNA is involved in the pathogenesis of MG. In our experiment, we extracted peripheral blood mononuclear cells (PBMCs) from MG patients and detected the expression of miR-181a and TRIM9 in PBMCs by qRT-PCR. In vitro experiments were conducted to explore the regulatory mechanism of miR-181a on target genes and its influence on inflammatory factors related to MG disease. Experimental autoimmune myasthenia gravis (EAMG) model mice are established, and the effects of miR-181a on EAMG symptoms and inflammatory factors are explored through in vivo experiments. According to a total of 40 EAMG mice that were successfully modeled, all EAMG mice showed symptoms of muscle weakness; their diet was reduced; their weight gain was slow; and even weight loss occurred. In MG patients and EAMG mice, the expression of miR-181a was low and TRIM9 was highly expressed. Bioinformatics website and dual-luciferase report analysis of miR-181a had a targeting relationship with TRIM9, and miR-181a could target the expression of TRIM9. After upregulating miR-181a or interfering with TRIM9, serum miR-181a in EAMG mice was significantly upregulated; TRIM9 was significantly downregulated; its clinical symptoms were reduced; and the expression of inflammatory factors was reduced. The study finally learned that miR-181a can reduce the level of MG inflammatory factors by targeting the expression of TRIM9 and has the effect of improving the symptoms of MG.

MiRNA-29a serves as a promising diagnostic biomarker in children with temporal lobe epilepsy and regulates seizure-induced cell death and inflammation in hippocampal neurons.

Temporal lobe epilepsy (TLE) in children is one of the most common refractory epilepsies. MicroRNAs (miRNAs) show abnormal expression in neurological disorders. The objective of this study was to determine changes in expression and the role of miR-29a in children with TLE. Sixty-five TLE patients and 70 normal controls were recruited. The levels of miR-29a were quantified using qRT-PCR. An in vitro TLE cell model was established using primary hippocampal cells cultured in magnesium-free medium. Cell viability, cell apoptosis and inflammatory cytokine concentrations were evaluated. The luciferase reporter assay was applied to confirm the target gene, HMGB1. A low level of MiR-29a expression was observed in the serum of children with TLE, which demonstrated a negative association with the concentration of serum TNF-α, IL-6, and IFN-γ. The level of MiR-29a demonstrated high specificity and sensitivity in children with TLE. A low level of expression of miR-29a was also detected in the TLE cell model. MiR-29a over-expression reversed the decreased cell viability induced by TLE, and alleviated cell apoptosis. Release of TNF-α, IL-6, and IFN-γ induced by TLE was also inhibited by miR-29a over-expression. HMGB1, which was downregulated in the serum of TLE patients, was shown to be a target gene of miR-29a, and negatively correlated with miR-29a level. The downregulation of serum miR-29a may serve as a non-invasive diagnostic biomarker for children with TLE. MiR-29a may be involved in the pathogenesis of TLE through regulation of neuronal apoptosis and neuroinflammation via targeting HMGB1.

Clinical value of miR-135 and miR-20a combined with multi-detector computed tomography in the diagnosis of gastric cancer

BackgroundTo study the clinical value of miR-135 and miR-20a combined with multi-detector computed tomography (MDCT) in the diagnosis of gastric cancer (GC).MethodA total of 146 patients with GC admitted to our hospital from January 2017 to June 2019 were selected and enrolled in the GC group. Another 103 patients with gastritis received in the same period were selected for the non-GC group. Besides, 95 healthy subjects who received physical examination in our hospital were selected into the healthy control group. Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression of serum miR-135 and miR-20a for each group. MDCT was used for detecting the clinical staging map of the enrolled patients. Pearson’s correlation analysis was used to analyze the correlation between serum miR-135 and miR-20a in patients with GC. The receiver operating characteristic (ROC) curve was drawn to analyze value of miR-135 and miR-20a in the diagnosis of GC.ResultsCompared with non-GC group and healthy control group, the levels of serum miR-135 and miR-20a increased significantly in the GC group, while no significant difference was found between non-GC group and healthy control group (P > 0.05). Analysis of the relationship with clinical characteristics showed that the expression of serum miR-135 and miR-20a in the GC group was significantly correlated with the progression of GC, TNM stage, degrees of differentiation, status of lymph node metastasis, and distant metastasis (P < 0.01). Pearson’s correlation analysis results showed positive correlations between miR-135 and miR-20a (r = 0.634, P = 0.000). The ROC analysis results showed that the optimal diagnostic values of miR-135 and miR-20a for GC were 7.56 and 5.82 respectively. The area under the curve (AUC) was 0.873 and 0.793 respectively. The 95% confidence interval (CI) was 0.811-0.935 and 0.697-0.890 respectively. The sensitivity and specificity of miR-135 and miR-20a combined with MDCT in the diagnosis of GC were 90.41% and 93.20% respectively. The sensitivity of combined use was significantly higher than that of single detection (P < 0.01).ConclusionThere are high expression levels of serum miR-135 and miR-20a in patients with GC. A combined detection of miR-135 and miR-20a with MDCT can improve the diagnostic sensitivity of GC and improve the accuracy of the final diagnosis. Therefore, multiple combined detection is valuable in the diagnosis of GC.

Serum MiRNA-23a as a Diagnostic and Prognostic Biomarker of Hepatitis C Related Hepatocellular Carcinoma

Background: Micro-ribonucleic acids (MiRNAs) are small, non-coding RNA molecules which regulate gene expression. Several miRNAs including miR-23a were found to be frequently deregulated in hepatocellular carcinoma (HCC). Objective: This study aimed to evaluate serum miR-23a as a biomarker of hepatitis C related HCC. Methods: This study was conducted on 60 hepatitis C virus (HCV) infected patients (group I: without cirrhosis, group II with cirrhosis and group III with HCV associated HCC) and a control group of 20 healthy volunteers. All patients were submitted to history taking, clinical examination in addition to categorization and staging of HCC patients. Following extraction of RNA from serum samples, quantitative reverse transcription polymerase chain reaction (qRT-PCR) was performed. Calculation of serum miR-23a was done using the comparative cycle threshold (Ct) method (2–ΔΔCT). Results: Serum miR-23a levels (2-ΔΔCT) were significantly higher in cirrhotic and HCC patients compared to chronic hepatitis C patients (CHC). However, no significant difference was noted between cirrhotic and HCC patients. The sensitivity and specificity of miR-23a levels for discriminating HCC patients from cirrhotic patients were 55% and 65%, respectively. MiR-23a levels had sensitivity of 90% and specificity of 70% for discriminating metastatic from non-metastatic HCC patients. Conclusion: Higher miR-23a levels were detected among metastatic HCC patients than among those without metastasis. The sensitivity and specificity of miR-23a levels for discriminating HCC patients from cirrhotic patients were lower than those of alpha fetoprotein (AFP).

Investigation of the levels of circulating miR-29a, miR-122, sestrin 2 and inflammatory markers in obese children with/without type 2 diabetes: a case control study

AimThe present work investigated serum levels of miR-29a, miR-122 and sestrin2 in obese children with/without type-2-diabetes mellitus (T2DM), and their correlations with inflammatory, metabolic and anthropometric parameters.MethodsThe study included 298 children, divided into: G1 (control, n = 136), G2 (obese without diabetes, n = 90) and G3 (obese with T2DM, n = 72). Metabolic and anthropometric parameters, miR-29a, miR-122 relative expressions, and sestrin2, high sensitivity C-reactive protein (hsCRP), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) levels were measured by their specific methods. The data was processed and analyzed by SPSS V.26 using the corresponding tests. After testing the variables’ normality, Kruskal–Wallis one-way-ANOVA, Spearman correlations coefficient were used.ResultsSignificant higher serum miR-29a, miR-122, IL-6, hsCRP and TNF-α and lower sestrin2 levels were found in G2 and G3 than G1 and in G3 than G2 (p= > 0.001 for all). Especially in G3, miR-29a and miR-122 levels correlated positively while sestrin2 levels correlated negatively with waist circumference and BMI percentiles, serum levels of LDL-cholesterol, triacylglycerol, total cholesterol, HbA1c%, glucose, insulin, c-peptide, homeostatic model assessment-insulin resistance (HOMA-IR), IL-6, hsCRP and TNF-α.ConclusionThe change in the serum miR-29a, miR-122 and sestrin2 levels in obese children with/without T2DM may suggest a possible role of these biomarkers in the pathogenesis of childhood obesity and their accompanied complications e.g. inflammations and T2DM. Also, further studies are required to test drugs that antagonize the action miR-29a and miR-122 or upregulate sestrin2 in the management of these cases.

Serum miR-27a is a biomarker for the prognosis of non-small cell lung cancer patients receiving chemotherapy.

BackgroundLung cancer has a high incidence and a 5-year survival rate of less than 15%. Non-small cell lung cancer (NSCLC) accounts for approximately 85% of lung cancer cases. Chemotherapy and immunotherapy are the most frequently used alternative treatments for patients with advanced-stage NSCLC in whom surgery failed. Previous studies have suggested that miR-27a is involved in cancer development and progression. The purpose of this study was to investigate the clinical value of miR-27a in the prognosis of NSCLC patients after chemotherapy.MethodsFlow cytometry was used to detect the apoptosis rate of SPC-A1 cells treated with optical cisplatin at different times. Simultaneously, the expression of miR-27a in supernatants and cells was detected. Fifty-two newly diagnosed NSCLC patients were recruited. All patients received gemcitabine and cisplatin as first-line chemotherapy and docetaxel as second-line chemotherapy. At the end of every chemotherapy cycle, a therapeutic evaluation was performed according to the RECIST criteria. The expression of serum miR-27a was detected in each cycle.ResultsAfter treatment with 2.5 µg/mL cisplatin, the apoptosis rates of SPC-A1 cells were significantly greater than those of the paired untreated control groups at 12, 24, 48 and 72 h. The expression of miR-27a in supernatants and cells was also consistent with the apoptosis rate and changed a time-dependent manner. The chi-square test showed that an increase in miR-27a after chemotherapy was more common in patients who achieved partial response (PR) than in those who achieved no response (NR) (61.5% vs. 30.8%, P=0.026). Kaplan-Meier survival analysis indicated that patients with decreased miR-27a levels had poorer outcomes than those with increased miR-27a levels (P<0.05). Furthermore, dynamic changes in serum miR-27a with a gradual increasing trend during chemotherapy predicted a good prognosis.ConclusionsCollectively, our results suggest that miR-27a is involved in the apoptosis of lung cancer cells and that serum miR-27a levels are related to the prognosis of NSCLC patients. The expression levels of miR-27a in the serum may be an independent predictor for the prognosis of NSCLC.

Expression and Significance of Serum Mir-146a and Mir-30e in Patients with Secondary Hemophagocytic Lymphohistiocytosis

ObjectiveHemophagocytic lymphohistiocytosis (HLH) is a clinical syndrome which is caused by uncontrolled activation and proliferation of CD8+ T cell , and abundant releasing of in inflammatory cytokines. It has been confirmed that miR-146a is widely participate in the regulation of intensity of inflammatory response, while miR-30e is related to the regulation of NK cell cytotoxicity. In this study, RT-PCR was used to detect the expression of serum miR-146a and miR-30e in patients with secondary HLH (sHLH). Their correlation with clinical data, and diagnosis and prognosis value in sHLH patients were explored. In addition, their relation with the proportion of cytotoxic lymphocyte subsets and the expression of perforin were explored.MethodsA total of 35 patients with sHLH and 10 healthy controls were enrolled in this study. Serum expression levels of miR-146a and miR-30e were detected by using RT-PCR. The expression of perforin protein oncytotoxic lymphocytes and the proportion of cytotoxic lymphocyte subsets were tested by flow cytometry. The clinical data of sHLH patients were collected.Results(1) The serum expression level of miR-146a in the newly diagnosed sHLH group was significantly lower than that of healthy control group (P <0.001); the serum expression level of miR-30e in the newly diagnosed sHLH group was significantly lower than that of healthy control group (P <0.001). (2) The serum expression level of miR-146a and miR-30e in the lymphoma-associated HLH group, the infection- associated HLH group, and the autoimmune-associated HLH group were compared group by group, however no significant difference was found (P >0.05). (3) ROC analysis revealed that the serum expression level of miR-146a can effectively identify sHLH patients from healthy people, with area under the curve (AUC) was 0.931; the serum expression level of miR-30e can effectively identify sHLH patients from healthy people, with area under the curve (AUC) was 0.914. (4) Correlation analysis was applied and no correlation was found between the serum expression level of miR-146a and miR-30e, and Age, WBC, HB, PLT, SF, ALB, ALT, AST, LDH, FIB, TG, ESR, sCD25, CRP level in the newly diagnosed sHLH (P >0.05). (5) According to the median of serum expression level, sHLH patients were divided in to Low expression group and High expression group. OS analysis was applied and revealed that there was no significance difference between the Low miR-146a expression group and the High miR-146a expression group (P >0.05); meanwhile there was no significance difference between the low miR-30e expression group and the high miR-30e expression group (P >0.05). However, sHLH patients were divided in to Low miR-30e/miR-146a group and High miR-30e/miR-146a group according to the median of miR-30e/miR-146a. The OS of Low miR-30e/miR-146a group was significantly longer than High miR-30e /miR-146a group (P=0.022). (6)The proportion of CD8+ T cell in Low miR-146a group was significantly lower than that in High miR-146a group (P = 0.016). The serum exression level of miR-146a was corelated with the proportion of CD8+ T cell (r=0.734, P = 0.007). However, the proportion of cytotoxic lymphocyte subsets in Low miR-146a group has no significant difference with that in Low miR-146a group (P >0.05); the serum exression level of miR-30e has no corelation with the proportion of cytotoxic lymphocyte subsets (P >0.05). (7) The expression of perforin in cytotoxic lymphocyte subsets in Low miR-30e group has no significant difference with that in Low miR-30e group (P >0.05); the serum exression level of miR-30e has no corelation with the expression of perforin in cytotoxic lymphocyte subsets (P >0.05).ConclusionThe serum expression level of miR-146a and miR-30e in the newly diagnosed sHLH group was significantly decreased, which is important in the differential diagnosis of sHLH. Individuals with lower serum expression level of miR-146a and miR-30e were more likely to be sHLH patients. The ratio of miR-30e and miR-146a (miR-30e /miR-146a) can indicate the prognosis of sHLH patients. The prognosis of sHLH patients with lower miR-30e /miR-146a was better than that of sHLH patients with higher miR-30e /miR-146a. The serum exression level of miR-146a was corelated with the proportion of CD8+ T cell. The serum exression level of miR-146a was corelated with the proportion of CD8+ T cell. Therefore, miR-146a may play a role in the deficiency of CD8+ T cell in the pathogenesis of sHLH. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Association between miR-146a and Tumor Necrosis Factor Alpha (TNF-α) in Stable Coronary Artery Disease.

Background and Objectives: Tumor necrosis factor alpha (TNF-α) is proatherogenic and associated with the risk of acute ischemic events, although the mechanisms that regulate TNF-α expression in stable coronary artery disease (SCAD) are not fully understood. We investigated whether metabolic, inflammatory, and epigenetic (microRNA (miRNA)) markers are associated with TNF-α expression in SCAD. Materials and Methods: Patients with SCAD were prospectively recruited and their metabolic and inflammatory profiles were assessed. TNF-α levels were assessed using an enzyme-linked immunosorbent assay. The relative expression of six circulating miRNAs associated with the regulation of inflammation and/or atherosclerosis was determined. Results: Of the 24 included patients with the mean age of 65 (9) years, 88% were male, and 54% were diabetic. The TNF-α levels were (median (interquartile range)) 1.0 (0.7–1.1) pg/mL. The percentage of glycosylated hemoglobin (r = 0.418, p = 0.042), serum triglyceride levels (r = 0.429, p = 0.037), and C-reactive protein levels (r = 0.407, p = 0.048) were positively correlated with TNF-α levels. Of the candidate miRNAs, miR-146a expression levels were negatively correlated with TNF-α levels (as indicated by r = 0.500, p = 0.035 for correlation between delta cycle threshold (ΔCt) miR-146a and TNF-α levels). In multivariate analysis, serum triglyceride levels and miR-146a expression levels were independently associated with TNF-α levels. miR-146 expression levels were not associated with metabolic or other inflammatory parameters and were negatively correlated with the number of coronary vessels with obstructive disease (as indicated by r = 0.556, p = 0.017 for correlation between ΔCt miR-146a and number of diseased vessels). Conclusions: miR-146a expression levels were negatively correlated with TNF-α levels in patients with SCAD, irrespective of other metabolic or inflammatory markers, and with the severity of coronary artery disease. The results add to the knowledge on the role of miR-146a in TNF-α-based inflammation in SCAD and support future research on the potential therapeutic use of miR-146a in such a clinical scenario.

Circulating Levels of MicroRNAs in Hypertrophic Cardiomyopathy: The Relationship With Left Ventricular Hypertrophy, Left Atrial Dilatation and Ventricular Depolarisation-Repolarisation Parameters

MicroRNAs are small, endogenous, non-coding RNAs that regulate the expression of many genes. It has recently been shown that circulating microRNAs may be biomarkers of hypertrophy and fibrosis in patients with hypertrophic cardiomyopathy (HCM). To determine whether circulating levels of microRNAs involved in HCM are associated with electrocardiographic and echocardiographic parameters. This study enrolled 20 patients with familial HCM and 20 blood donors. Peripheral serum levels of miR-29a-3p, miR-199a-5p and miR-451a were assessed by quantitative real-time polymerase chain reaction and compared with levels in the control group. Whether circulating levels of miRNAs in HCM patients correlated with electrocardiographic and echocardiographic parameters was also assessed. Median circulating levels of miR-29a and miR-451a were significantly higher in HCM than the control group. Median miR-199a levels did not differ between groups. However, circulating levels of miR-199a negatively correlated with corrected QT duration (Bazett formula). Median miR-29a levels positively correlated with QRS duration. In addition, circulating levels of miR-29a correlated with maximal wall thickness, left ventricular mass index and left atrial volume index. The data suggested that serum levels of miR-29a and miR-451a were significantly increased in HCM patients. As the circulating level of miR-29a correlated with QRS duration, left ventricular hypertrophy and left atrial dilatation, the serum miR-199a level negatively correlated with corrected QT duration. These miRNAs may be seen as potential biomarkers for further research in HCM pathophysiology.

Expression and Clinical Significance of miR-146a and Tumor Necrosis Factor Receptor-Associated Factor 6 (TRAF6) in Myasthenia Gravis Patient Serum.

Objective To investigate the expression and clinical significance of miR-146a and tumor necrosis factor receptor-associated factor 6 (TRAF6) in myasthenia gravis (MG) patient serum. Methods The serum of 52 patients with MG and 60 healthy individuals was collected in our hospital. The expression of miR-146a and TRAF6 in serum was measured by real-time PCR (RT-PCR). Comparison among serum miR-146a and TRAF6 mRNA group clinical characteristics and assorted expressions was done with the correlation among the two groups evaluated. Logistics regression was used to analyze the effect of miR-146a and TRAF6 mRNA on MG development with the ROC curve applied for an investigation into the diagnostic role of miR-146a and TRAF6 mRNA expression in MG development. Results miR-146a and TRAF6 mRNA were significantly increased in the patients with MG compared with the healthy controls. Significant differences were identified in respiratory muscle endurance, muscle weakness level, vital capacity, and maximal voluntary ventilation between the two groups. Additionally, correlation analysis has discovered a positive correlation between miR-146a and TRAF mRNA expression in patients with MG. miR-146a and TRAF6 mRNA are independent MG occurrence factors exhibited by multivariate analysis while areas under ROC curve (AUCs) of miR-146a and TRAF6 mRNA in MG diagnosis were established by ROC curve analysis with results being 0.782 and 0.703, correspondingly. Conclusion miR-146a and TRAF6 mRNA are highly expressed in MG patients and can affect MG occurrence. miR-146a is a suitable candidate marker for diagnosing MG.

Role of serum miR-21 and miR-92a in colorectal cancer diagnosis as novel molecular biomarkers

Background: Our study aimed to evaluate the clinical utility of detecting plasma microRNAs (miR-21 & miR-92a) for diagnosis of colorectal cancer patients and its relation to tumor staging. Patients and Methods: Quantitative real-time RT-PCR was applied to determine the relative expression level of miR-21 and miR-92a in serum. The sensitivity and specificity of these markers were evaluated by receiver operating characteristic (ROC) curve analysis. Final staging of colorectal cancer cases was assigned according to results of histopathologic examination of surgically resected specimens. Results: This study included 52 cases of colorectal cancer (CRC), 20 cases of precancerous colorectal lesions, and 20 healthy controls. Both Plasma miR-21 and Plasma miR-92a were significantly higher in CRC group compared to both the control group and precancerous group. Also, they were significantly higher in advanced CRC stages than early CRC stages. The sensitivity and specificity of miR-21 for discriminating CRC from controls were found to be 90.38% and 100.0%, respectively. However, for miR-92a, sensitivity and specificity were found to be 94.23% and 100.0%, respectively. For discriminating CRC cases from precancerous lesions, the sensitivity and specificity of miR-21 were found to be 75.08% and 95.0%, respectively. However, for miR-92a, sensitivity and specificity were found to be 80.77% and 100.0%, respectively. Conclusions: both plasma miR-21 and miR-92a have significant value for early detection of CRC as non-invasive screening molecular biomarkers with high sensitivity and specificity. They also help for differentiation between patients with benign and malignant colorectal lesions and those with early and advanced CRC.

Prognostic markers for severity, activity, and recurrence of alopecia areata

Background Alopecia areata (AA) is considered an autoimmune disorder caused by altered T-cell-mediated immunity. MicroRNAs (miR) are important translational regulators of genes in various tissues and biological processes involved in autoimmune responses and have been identified in autoimmune diseases such as rheumatoid arthritis and type I diabetes mellitus. However, little is known about their role in pathogenesis of AA.Objective To assess serum levels of miR-155 and miR-146a in patients with AA and to correlate them with different clinical variables. Patients and methods MiR-155 and miR-146a serum levels were identified by real-time PCR in 50 patients with AA and 50 healthy age-matched and sex-matched patients as controls. Disease severity in the patients was assessed by Severity of Alopecia Tool score. Results There was a statistically significant increase in serum miR-155 and miR-146a levels in patients with AA than in controls. A significant increase in serum levels of miR-155 was found in patients with active disease and recurrent lesions of AA, whereas significant increase in serum level of miR-146a was found in patients with recurrent disease only. Significant positive correlations were found between serum miR-155 and miR-146a levels and disease severity. Conclusion Serum levels of miR-155 and miR-146a seem to have an essential role in the etiopathogenesis of AA and could be markers for severity and early detection of recurrent AA. In addition, serum miR-155 could be a marker of activity in AA, whereas serum miR-146a could be a marker of multiplicity. Further understanding of the function and regulation of miR-155 and miR-146a could be of great value for the introduction of new therapeutic approaches for AA.

Diagnostic Value of Vaginal Microecology, Serum miR-18a, and PD-L1 for Identifying HPV-Positive Cervical Cancer.

Objective:We aimed to investigate the diagnostic value of the vaginal microecology, serum miR-18a, and programmed death ligand-1 (PD-L1) for human papillomavirus (HPV)-positive cervical cancer.Methods:Eighty-four patients with HPV-positive cervical cancer were assigned to the observation group, 107 HPV-positive patients without cervical cancer were assigned to the positive group, and 191 healthy women were assigned to the control group. Vaginal microecology and serum levels of miR-18a and PD-L1 on the surface of CD4+ and CD8+ T cells were compared among the 3 groups. The observation group was further divided into subgroups according to patients’ characteristics for comparison. The diagnostic value of miR-18a and PD-L1 for HPV-positive cervical cancer was investigated.Results:Women in the control group had better vaginal microecology and lower levels of miR-18a and PD-L1 than those in the observation and the positive groups (all P < 0.05). Compared with the positive group, the observation group had similar vaginal microecology (all P > 0.05) but higher levels of miR-18a and PD-L1 (all P < 0.05). Moreover, the patients at stage III had higher levels of miR-18a and PD-L1 than those at stage I and II (all P < 0.05). The values of area under the curve for miR-18a and PD-L1 in the diagnosis of HPV-positive cervical cancer were over 0.8 (all P < 0.001).Conclusion:Patients with HPV-positive cervical cancer have vaginal microbial dysbiosis and high serum levels of miR-18a and PD-L1. miR-18a and PD-L1 have diagnostic value for identifying HPV-positive cervical cancer.

MiR-18a increases insulin sensitivity by inhibiting PTEN.

The miR-17-92 cluster (miR-17, miR-18a, miR-19a, miR-20a, miR-19b-1 and miR-92a) contributes to the occurrence and development of various diseases by inhibiting multiple target genes. Here, we explored the effects of miR-18a on insulin sensitivity. Quantitative real-time PCR indicated that serum miR-18a levels were lower in type 2 diabetes mellitus patients than in healthy controls, suggesting that miR-18a may influence blood glucose levels. Global overexpression of miR-18a in transgenic mice increased their glucose tolerance and insulin sensitivity, while it reduced expression of the phosphatase and tensin homolog deleted on chromosome ten (PTEN) in their skeletal muscle and adipose tissue. Western blotting indicated that overexpressing miR-18a in 3T3-L1 and C2C12 cells enhanced insulin-stimulated AKT phosphorylation and suppressed PTEN expression, while inhibiting miR-18a had the opposite effects. These results suggest that miR-18a improves insulin sensitivity by downregulating PTEN. This makes miR-18a a potentially useful target for the treatment of diabetes mellitus in the future.

Targeting miR-18a sensitizes chondrocytes to anticytokine therapy to prevent osteoarthritis progression

Inflammation participates in the development of OA and targeting inflammatory signaling pathways is a potential strategy for OA treatment. IL-1β is one of the most important inflammatory factors to trigger the activation of NF-κB signaling and accelerate OA progression, whereas OA patients could hardly benefit from inhibiting IL-1β in clinic, suggesting the importance to further explore the details of OA inflammation. We here showed that expression of miR-18a in chondrocytes was specifically induced in response to IL-1β in vitro as well as in rat model of OA during which NF-κB signaling was involved, and that nuclear-translocated p65 directly upregulated miR-18a expression at transcriptional level. Further, increased miR-18a mediated hypertrophy of chondrocytes, resulting in OA degeneration, by targeting TGFβ1, SMAD2, and SMAD3 and subsequently leading to repression of TGF-β signaling. And the level of serum miR-18a was positively correlated to severity of OA. Interestingly, other than IL-1β, pro-inflammation cytokines involving TNFα could also remarkably upregulate miR-18a via activating NF-κB signaling and subsequently induce chondrocytes hypertrophy, suggesting a pivotal central role of miR-18a in inflammatory OA progression. Thus, our study revealed a novel convergence of NF-κB and TGF-β signaling mediated by miR-18a, and a novel mechanism underlying inflammation-regulated OA dependent of NF-κB/miR-18a/TGF-β axis. Notably, in vivo assay showed that targeting miR-18a sensitized OA chondrocytes to IL-1β inhibitor as targeting IL-1β and miR-18a simultaneously had much stronger inhibitory effects on OA progression than suppressing IL-1β alone. Therefore, the diagnostic and therapeutic potentials of miR-18a for OA were also revealed.

Serum miRNAs associated with tumor-promoting cytokines in non-small cell lung cancer.

Tumor-promoting cytokines are a cause of tumor progression; therefore, identifying key regulatory microRNAs (miRNAs) for controlling their production is important. The aim of this study is to identify promising miRNAs associated with tumor-promoting cytokines in non-small cell lung cancer (NSCLC). We identified circulating miRNAs from 16 published miRNA profiles. The selected miRNAs were validated in the serum of 32 NSCLC patients and compared with 33 patients with other lung diseases and 23 healthy persons using quantitative real-time PCR. The cytokine concentration was investigated using the enzyme-linked immunoassay in the same sample set, with clinical validation of the miRNAs. The correlation between miRNA expression and cytokine concentration was evaluated by Spearman's rank correlation. For consistent direction, one up-regulated miRNA (miR-145) was found in four studies, and seven miRNAs were reported in three studies. One miRNA (miR-20a) and four miRNAs (miR-25-3p, miR-223, let-7f, and miR-20b) were reported in six and five studies. However, their expression was inconsistent. In the clinical validation, serum miR-145 was significantly down-regulated, whereas serum miR-20a was significantly up-regulated in NSCLC, compared with controls. Regarding serum cytokine, all cytokines [vascular endothelial growth factor (VEGF), interleukin-6 (IL-6), and transforming growth factor β (TGF-β)], except tumor necrosis factor-α (TNF-α), had a higher level in NSCLC patients than controls. In addition, we found a moderate correlation between the TGF-β concentration and miR-20a (r = -0.537, p = 0.002) and miR-223 (r = 0.428, p = 0.015) and a weak correlation between the VEGF concentration with miR-20a (r = 0.376, p = 0.037) and miR-223 (r = -0.355, p = 0.046). MiR-145 and miR-20a are potential biomarkers for NSCLC. In addition, the regulation of tumor-promoting cytokine, through miR-20a and miR-223, might be a new therapeutic approach for lung cancer.

The value of serum microRNA-92a and microRNA-146a levels combined with pulmonary ultrasound score in predicting the severity and prognosis of acute respiratory distress syndrome

To investigate the value of serum microRNA-92a (miR-92a) and microRNA-146a (miR-146a) expression levels combined with lung ultrasound score (LUS) in predicting the severity and prognosis of acute respiratory distress syndrome (ARDS). 116 patients with ARDS admitted to Danzhou People's Hospital from January 2017 to March 2020 were enrolled. On the day of admission, the expression levels of serum miR-92a and miR-146a were detected by real-time fluorescent quantitative reverse transcript-polymerase chain reaction (RT-PCR), and pulmonary ultrasound examination was performed in 12 lung regions, with the total score as LUS score. The difference of each index was analyzed among the ARDS patients with different 28-day prognosis (survival group and death group) and different severity [mild group: 200 mmHg < oxygenation index (OI) ≤ 300 mmHg (1 mmHg = 0.133 kPa), moderate group: 100 mmHg < OI ≤ 200 mmHg, severe group: OI ≤ 100 mmHg]. Multivariate Logistic regression was used to analyze the risk factors of death in patients with ARDS. Receiver operating characteristic (ROC) curve was drawn to analyze the value of miR-92a and miR-146a combined with LUS score in predicting the death of patients with ARDS. 116 ARDS patients were included, 39 cases in the death group, 77 cases in the survival group; 20 cases in the mild group, 38 cases in the moderate group and 58 cases in the severe group. The expression levels of serum miR-92a, miR-146a and LUS score in the death group were significantly higher than those in the survival group [miR-92a (2-ΔΔCt): 3.75±1.64 vs. 2.10±0.78, miR-146a (2-ΔΔCt): 1.93±0.72 vs. 0.76±0.20,LUS score: 25.80±4.75 vs. 13.40±3.60, all P < 0.01]. With the aggravation of ARDS patients, the expression levels of serum miR-92a and miR-146a and LUS score gradually increased (F values were 8.115, 6.740 and 6.216 respectively, all P < 0.01). The expression levels of serum miR-92a, miR-146a and LUS score in severe group were significantly higher than those in the moderate group and mild group [miR-92a (2-ΔΔCt): 3.65±1.62 vs. 2.87±1.16, 1.94±0.68; miR-146a (2-ΔΔCt): 1.85±0.58 vs. 1.30±0.51, 0.68±0.17; LUS score: 24.15±4.65 vs. 18.60±4.20, 12.20±3.15, all P < 0.01]. Multivariate Logistic regression analysis showed that low OI [odds ratio (OR) = 2.748, 95% confidence interval (95%CI) was 1.913-6.225, P = 0.024], high LUS score (OR = 1.685, 95%CI was 1.183-2.758, P = 0.016), high expression levels of serum miR-92a (OR = 2.560, 95%CI was 1.806-5.627, P < 0.001) and miR-146a (OR = 1.984, 95%CI was 1.375-3.816, P = 0.008) were independent risk factors for the death of ARDS patients. ROC curve analysis showed that the area under ROC curve (AUC) of patients with ARDS predicted by miR-92a and miR-146a combined with LUS score was significantly higher than that predicted by the three alone (0.918 vs. 0.842, 0.825, 0.807, all P < 0.01), and the sensitivity (94.0%) and specificity (85.2%) were higher. The expression levels of serum miR-92a, miR-146a and LUS score are related to the severity and prognosis of the patients with ARDS, and the combination of the three indicators has better value in predicting the prognosis of the patients with ARDS.

Potential Value of miR-23a for Discriminating Neuromyelitis Optica Spectrum Disorder from Multiple Sclerosis

Until now, no laboratory test or test set can guarantee the diagnosis of multiple sclerosis (MS) at early disease stages, and the disease symptoms may interfere with many other disease conditions. Analyzing the expression of circulating miRNAs may provide a useful approach for early and differential MS diagnosis. The main objective is assessment of the potential of serum miR-23a, miR-155, and miR-572 to differentiate between MS and other neuroinflammatory diseases. Serum miRNAs were obtained from 37 MS patients and 25 healthy age-matched controls, along with patients with neuromyelitis optica spectrum disorder (NMOSD) [n = 13] and neuropsychiatric systemic lupus erythematosus (NPSLE) [n = 10]. miRNA expression levels were analyzed using real-time polymerase chain reaction (PCR) and pairwise comparisons were made to reveal the diagnostic/distinguishing potential of the analyzed miRNAs. In the study cohort, the three investigated miRNAs failed to display significant dysregulation in MS patients. However, they could significantly discriminate patients with NMOSD and NPSLE [median (IQR): 8.1 (6.1-9.2) and 8.8 (7.9-9.7) for miR-23a, 7.5 (5.3-8.3) and 8.0 (7.5-9.5) for miR-155 and 6.9 (5.0-8.8) and 6.4 (5.3-8.8) for miR-572 in NMOSD and NPSLE, respectively] from healthy subjects [median (IQR): 3.4 (1.5-4.3), 3.1 (1.1-5.6) and 3.5 (1.7-5.6) for miR-23a, miR-155 and miR-572, respectively], with area under the curve (AUC) ≤0.8. Remarkably, miR-23a has been emerging as a prospective biomarker for differentiation of MS from NMOSD as well as NPSLE (AUC<0.9). The miRNA combined use contributed to enhanced diagnostic and discriminatory performance in the study groups. Certain miRNA expression levels would contribute to discriminating MS from other neuroinflammatory diseases.

Serum miR-29a/b expression in gestational diabetes mellitus and its influence on prognosis evaluation.

ObjectiveTo evaluate serum microRNA (miR)-29a/b expression in gestational diabetes mellitus (GDM) and its influence on neonatal prognosis.MethodsThis was a retrospective study including 68 pregnant women with GDM (GDM group) and 55 healthy pregnant women of similar age range and gestation period (healthy group).ResultsThe area under the curve was 0.829 for the diagnosis of GDM using serum miR-29a expression, 0.857 for diagnosis using serum miR-29b expression, and 0.944 for combined diagnosis (using both miR-29a and miR-29b). The fasting insulin (FINS) level of the GDM group was significantly lower than that of the healthy group; levels of fasting plasma glucose (FPG), 2-h plasma glucose (2hPG), and glycated hemoglobin (HbA1c) were significantly higher in the GDM group than in the healthy group. Both miR-29a and miR-29b were positively correlated with FINS levels and negatively correlated with FPG, 2hPG, and HbA1c levels. Serum miR-29a/b expression in pregnant women with GDM was not correlated with neonatal weight, premature delivery, or asphyxia but was correlated with pathologic jaundice.ConclusionsSerum miR-29a/b expression was downregulated in pregnant women with GDM and correlated with neonatal pathologic jaundice, showing good individual (miR-29a or miR-29b) diagnostic value and excellent combined (miR-29a and miR-29b) diagnostic value.