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The association between microRNA (miRNA) and various diseases has been established; miRNAs have the potential to be biomarkers for these diseases. Nevertheless, the challenge of correctly quantifying an miRNA arises from its low abundance and a high degree of family homology. Therefore, in the present study, we devised a chemiluminescence (CL) detection method for miRNAs, known as the hybridization chain reaction (HCR)-CL, utilizing the enzyme-free signal amplification technology of HCR. The proposed methodology obviates the need for temperature conversion and offers a straightforward procedure owing to the absence of enzymatic participation, and the lumino-H2O2-mediated CL reaction occurs at a high rate. The technique successfully detected 2.5 amol of the target analyte and 50 amol of miR-146b in a 1% concentration of human serum. In summary, the method developed in this study is characterized by its ease of operation, cost-effectiveness, remarkable analytical prowess, and ability to detect miRNA without the need for total RNA extraction from serum samples. This method is expected to be widely used for biological sample testing in clinical settings. [ABSTRACT FROM AUTHOR]

Introduction: Samples classified as indeterminate correspond to 10-20% of cytologies obtained by fine needle biopsy of thyroid nodules, preventing an adequate distinction between benign and malignant lesions and leading to diagnostic thyroidectomies that often prove unnecessary, as most cases are benign. Furthermore, although the vastmajority of patients with differentiated thyroid cancer (DTC) have such a good prognosis that active surveillance is permitted as an initial therapeutic option, relapses are not rare, and a non-negligible number of patients experience poor outcomes. MicroRNAs(miR)emergeaspotentialbio markers capable of helping to define more precise management of patients in all these situations. Methods: Aiming to investigate the clinical utility of miR-146b-5p in the diagnostic of thyroid nodules and evaluating its prognostic potential in a realworld setting, we studied 89 thyroid nodule samples, correlating miR-146b-5p expression with clinical tools such as the 8th edition from the American Joint Committee on Cancer (AJCC/UICC) and the American Thyroid Association Guideline Stratification Systems for the rate of recurrence (RR). Results: miR-146b-5p expression levels distinguished benign from malignant thyroid FNA samples (p< 0.0001). For indeterminate nodules, overexpression of miR-146b-5p with a cut-off of 0.497 was able to diagnose malignancy with a 90% accuracy; specificity=87.5%; sensitivity=100%. An increased expression of miR-146b-5p was associated with greater RR (p=0.015). A cut-off of 2.21 identified cases with more vascular involvement (p=0.013) and a cut-off of 2.420 was associated with a more advanced TNM stage (p-value=0.047). Discussion: We demonstrated that miR-146b5p expression in FNA samples is able to differentiate benign frommalignant indeterminate nodules and is associatedwith an increased risk of recurrence andmortality, suggesting that this single miRNAmay be a useful diagnostic and prognostic marker in the personalized management of DTC patients. [ABSTRACT FROM AUTHOR]

Introduction: The repertoire of microRNAs (miRNAs) in thyroid carcinomas starts to be elucidated. Among differentiated thyroid carcinomas (DTCs), papillary thyroid carcinoma (PTC) is the most frequent. The assessment of miRNAs expression may contribute to refine the pre-surgical diagnosis in order to obtain a personalized and more effective treatment for patients. Aims: This study aims to evaluate (1) the miRNAs in a series of DTCs, and their association with the presence of selected genetic mutations in order to improve diagnosis and predict the biologic behavior of DTC/PTC. (2) The reliability of molecular tests in Ultrasound-guided Fine Needle Aspiration Cytology (US-FNAC) for a more precise preoperative diagnosis. Material and Methods: This series includes 176 samples (98 cytology and 78 histology samples) obtained from 106 patients submitted to surgery, including 13 benign lesions (controls) and 93 DTCs (cases). The microRNA expression was assessed for miR-146b, miR-221, miR-222, and miR-15a through quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR). The results were analyzed by the 2−ΔΔCT method, using miR16 as an endogenous control. Regarding PTC diagnosis, the discriminative ability of miRNAs expression was assessed by the area under the Receiver Operating Characteristic Curve (AUC). In PTCs, the association of miRNAs expression, clinicopathological features, and genetic mutations (BRAF, RAS, and TERTp) was evaluated. Results/Discussion: All the analyzed miRNAs presented a tendency to be overexpressed in DTCs/PTCs when compared with benign lesions, both in cytology and histology samples. In cytology, miRNAs expression levels were higher in malignant tumors than in benign tumors. In histology, the discriminative abilities regarding PTC diagnosis were as follows: miR-146b (AUC 0.94, 95% CI 0.87–1), miR-221 (AUC 0.79, 95% CI 0.68–0.9), miR-222 (AUC 0.76, 95% CI 0.63–0.89), and miR-15a (AUC 0.85, 95% CI 0.74–0.97). miR-146b showed 89% sensitivity (se) and 87% specificity (sp); miR-221 se = 68.4, sp = 90; miR-222 se = 73, sp = 70; and mi-R15a se = 72, sp = 80. MicroRNAs were associated with worst-prognosis clinicopathological characteristics in PTCs (p < 0.05), particularly for miR-222. Our data reveal a significant association between higher expression levels of miR-146b, miR-221, and miR-222 in the presence of the BRAF mutation (p < 0.001) and miR-146b (p = 0.016) and miR-221 (p = 0.010) with the RAS mutation, suggesting an interplay of these mutations with miRNAs expression. Despite this study having a relatively small sample size, overexpression of miRNAs in cytology may contribute to a more precise preoperative diagnosis. The miRNAs presented a good discriminative ability in PTC diagnosis. The association between the miRNAs expression profile and genetic alterations can be advantageous for an accurate diagnosis of DTCs/PTCs in FNAC. [ABSTRACT FROM AUTHOR]

IntroductionSamples classified as indeterminate correspond to 10-20% of cytologies obtained by fine needle biopsy of thyroid nodules, preventing an adequate distinction between benign and malignant lesions and leading to diagnostic thyroidectomies that often prove unnecessary, as most cases are benign. Furthermore, although the vast majority of patients with differentiated thyroid cancer (DTC) have such a good prognosis that active surveillance is permitted as an initial therapeutic option, relapses are not rare, and a non-negligible number of patients experience poor outcomes. MicroRNAs (miR) emerge as potential biomarkers capable of helping to define more precise management of patients in all these situations.MethodsAiming to investigate the clinical utility of miR-146b-5p in the diagnostic of thyroid nodules and evaluating its prognostic potential in a realworld setting, we studied 89 thyroid nodule samples, correlating miR-146b-5p expression with clinical tools such as the 8th edition from the American Joint Committee on Cancer (AJCC/UICC) and the American Thyroid Association Guideline Stratification Systems for the rate of recurrence (RR).ResultsmiR-146b-5p expression levels distinguished benign from malignant thyroid FNA samples (p< 0.0001). For indeterminate nodules, overexpression of miR-146b-5p with a cut-off of 0.497 was able to diagnose malignancy with a 90% accuracy; specificity=87.5%; sensitivity=100%. An increased expression of miR-146b-5p was associated with greater RR (p=0.015). A cut-off of 2.21 identified cases with more vascular involvement (p=0.013) and a cut-off of 2.420 was associated with a more advanced TNM stage (p-value=0.047).DiscussionWe demonstrated that miR-146b5p expression in FNA samples is able to differentiate benign from malignant indeterminate nodules and is associated with an increased risk of recurrence and mortality, suggesting that this single miRNA may be a useful diagnostic and prognostic marker in the personalized management of DTC patients.

In the face of the elevated incidence and mortality rate of septic shock in the ICU, this retrospective study seeks to investigate the indicative and predictive value of high-mobility group box 1 (HMGB1) and miR-146b in patients with septic shock. Quantitative RTPCR was employed in this study to quantify the HMGB1 and miR-146b levels in plasma samples obtained from the patient group and healthy controls. The investigation involved the comparison between the two groups and tracking changes in the patient group over time. The finding revealed that upon admission, the patient group exhibited markedly elevated relative expression levels of HMGB1, which subsequently decreased over time. Conversely, the patient group displayed significantly reduced relative expression levels of miR-146b upon admission, which subsequently increased over time compared to the control group. Receiver operating characteristic (ROC) curves showed good predictive value for HMGB1 and miR-146b. The experimental results suggest that HMGB1 and miR-146b serve as valuable and convenient biomarkers for evaluating the severity of septic shock and predicting mortality. Additionally, it is proposed that serum miR-146b may be inducible and potentially exerts a negative regulatory effect on the expression of HMGB1. [ABSTRACT FROM AUTHOR]

The objective was to identify, through in silico analysis, the genes to which miR-146a, miR-146b, and miR-155 bind and to analyze the metabolic pathways in which they participate during tuberculosis infection. For the analysis, it was used: miRBase, UniProtKB, TargetScan Human, miRDB, and miRTarBase. miR-146a interacts with or binds to genes important in cell adhesion and the process of phagocytosis (CLDN16 and ATP6V1C2, respectively) (P< 0.05); this interaction could have important implications in the pathogenesis of tuberculosis or related diseases. The results of this work suggest that the activation of specific molecular mechanisms in response to tuberculosis is regulated by miR-146a, miR-146b, and miR-155. The genes with which miR-146a and miR-155 interact or bind are involved in the immune response and cellular processes essential during tuberculosis infection. [ABSTRACT FROM AUTHOR]

Purpose: MicroRNA-146b (miR-146b) alleviates experimental colitis in mice by mediating macrophage polarization and the release of inflammatory factors. Our goals were to evaluate the antitumor efficacy of miR-146b in colorectal cancer (CRC) and to investigate the underlying mechanisms. Methods: We used murine models of CRC to evaluate whether miR-146b influenced the progression of tumors independent of tumor-associated macrophages (TAMs). RNA immunoprecipitation, N6-methyladenosine (m6A) RNA immunoprecipitation and in vitro pri-miRNA processing assays were conducted to examine whether m6A mediates the maturation of pri-miR-146b/miR-146b. In a series of in vitro and in vivo experiments, we further defined the molecular mechanisms of methyltransferase-like 3 (METTL3)/miR-146b-mediated antitumor immunity and its efficacy in combination with anti-PD-1 immunotherapy. Results: We found that miR-146b deletion supported tumor progression by increasing the number of alternatively activated (M2) TAMs. Mechanistically, the m6A-related "writer" protein METTL3 and "reader" protein HNRNPA2B1 controlled miR-146b maturation by regulating the m6A modification region of pri-miR-146b. Furthermore, miR-146b deletion promoted the polarization of M2-TAMs by enhancing phosphoinositide 3-kinase (PI3K)/AKT signaling, and this effect was mediated by the class IA PI3K catalytic subunit p110β, which reduced T cell infiltration, aggravated immunosuppression and ultimately promoted tumor progression. METTL3 knockdown or miR-146b deletion induced programmed death ligand 1 (PD-L1) production via the p110β/PI3K/AKT pathway in TAMs and consequently augmented the antitumor activity of anti-PD-1 immunotherapy. Conclusions: The maturation of pri-miR-146b is m6A-dependent, and miR-146b deletion-mediated TAM differentiation promotes the development of CRC by activating the PI3K/AKT pathway, which induces upregulation of PD-L1 expression, inhibits T cell infiltration into the TME and enhances the antitumor activity of anti-PD-1 immunotherapy. The findings reveal that targeting miR-146b can serve as an adjuvant to anti-PD-1 immunotherapy. [ABSTRACT FROM AUTHOR]

Spinal cord injury (SCI) is an incurable central nervous system impairment that lack of efficient treatment. Exosomes derived from mesenchymal stem cells (MSCs) are widely applied in disease treatment. This work aimed to determine the promising therapeutic effects of MSC-derived exosomal miRNA146b on SCI. A rat spinal cord injury (SCI) model and lipopolysaccharide (LPS)-induced PC12 cell model were established. Exosomes were extracted from human umbilical cord mesenchymal stem cells (hUCMSCs). The identification of exosomes was performed by using transmission electronic microscope (TEM) and nanoparticle tracking analysis (NTA). Hematoxylin and eosin (HE) staining and TUNEL assay were performed to assess tissue damage and apoptosis, respectively. ELISA was performed to detect levels of inflammatory cytokines. Cell viability was checked by cell counting kit 8 (CCK-8). Gene expression and protein levels were detected by qPCR and western blotting assay. The interaction between miR-146 b and Toll-like receptor 4 (TLR4) was assessed by luciferase reporter gene assay. The hUCMSC-derived exosomes could notably alleviate the spinal cord injury and cell apoptosis. The exosomal miR-146 b treatment suppressed the release of IL-1 β, IL-6, and TNFα. The miR-146 b suppressed the expression of TLR4, directly interact with the 3′-untranslated region (3′UTR) of TLR4, and inactivated the nuclear factor κB (NF-κB) signaling. The hUCMSCs-derived exosomal miR-146 b protects neurons from spinal cord injury through targeting the TLR4 and inactivating the NF-κB signaling. Our findings supported the application of hUCMSCs-derived exosomal miR-146 b for the protection of SCI.

Multiple sclerosis (MS) is a central nervous disorder caused by several factors. Studies have recently shown that non-coding RNA such as miRNA could participate in MS initiation, progression, and active lesion. This study aims to theoretically analyze the potential impact of single-nucleotide polymorphisms (SNPs) on mir-21 and mir-146a/b, which has been previously demonstrated as MS microRNA signature. To fulfill this purpose, the SNPs were investigated for functionality through several online tools, including miRNA-SNP, SNP2-TFBS, RBP-Var, and RNAfold. Furthermore, SNPs of miR-21 and miR-146a/b that exist in pre-miRNA, mature miRNA, and promoter area were extracted; moreover, miRNA and RNA-binding protein interactions were analyzed. This article presented a list of validated SNPs that could affect the expression or function of miR-21 and miR-146a/b for the future practical study of MS and active lesions. [ABSTRACT FROM AUTHOR]

The miR-146 family consists of two microRNAs (miRNAs), miR-146a and miR-146b, which are both known to suppress a variety of immune responses. Here in this study, we show that miR-146b is abundantly expressed in neuronal cells, while miR-146a is mainly expressed in microglia and astroglia of adult mice. Accordingly, miR-146b deficient (Mir146b-/-) mice exhibited anxiety-like behaviors and enhanced cognition. Characterization of cellular composition of Mir146b-/- mice using flow cytometry revealed an increased number of neurons and a decreased abundancy of astroglia in the hippocampus and frontal cortex, whereas microglia abundancy remained unchanged. Immunohistochemistry showed a higher density of neurons in the frontal cortex of Mir146b-/- mice, enhanced hippocampal neurogenesis as evidenced by an increased proliferation, and survival of newly generated cells with enhanced maturation into neuronal phenotype. No microglial activation or signs of neuroinflammation were observed in Mir146b-/- mice. Further analysis demonstrated that miR-146b deficiency is associated with elevated expression of glial cell line-derived neurotrophic factor (Gdnf) mRNA in the hippocampus, which might be at least in part responsible for the observed neuronal expansion and the behavioral phenotype. This hypothesis is partially supported by the positive correlation between performance of mice in the object recognition test and Gdnf mRNA expression in Mir146b-/- mice. Together, these results show the distinct function of miR-146b in controlling behaviors and provide new insights in understanding cell-specific function of miR-146b in the neuronal and astroglial organization of the mouse brain. [ABSTRACT FROM AUTHOR]

Epidermal growth factor receptor (EGFR) signalling and the interleukin-6 (IL-6)/signal transducer and activator of transcription 3 (STAT3) are aberrantly activated in ovarian cancer. However, inhibition of EGFR signalling in ovarian cancer patients resulted in a disappointing clinical benefit. In this study, we found that EGFR could activate IL-6-STAT3 pathway in ovarian cancer cells. However, we also demonstrated that EGFR knockdown could increase STAT3 phosphorylation in HO8910 and OVCAR-3 ovarian cancer cells. Interestingly, we further demonstrated that the non-coding RNA miR-146b could simultaneously block both the EGFR and IL-6-STAT3 pathways. Finally, our data demonstrated that miR-146b overexpression resulted in a greater suppression of cell migration than STAT3 pathway inhibition alone.These results suggest a complex and heterogeneous role of EGFR in ovarian cancer. Combined blockade of EGFR and IL-6-STAT3 pathways by miR-146b might be a strategy for improving the clinical benefit of targeting the EGFR pathway in ovarian cancer patients in the future.

Epidermal growth factor receptor (EGFR) signalling and the interleukin-6 (IL-6)/signal transducer and activator of transcription 3 (STAT3) are aberrantly activated in ovarian cancer. However, inhibition of EGFR signalling in ovarian cancer patients resulted in a disappointing clinical benefit. In this study, we found that EGFR could activate IL-6-STAT3 pathway in ovarian cancer cells. However, we also demonstrated that EGFR knockdown could increase STAT3 phosphorylation in HO8910 and OVCAR-3 ovarian cancer cells. Interestingly, we further demonstrated that the non-coding RNA miR-146b could simultaneously block both the EGFR and IL-6-STAT3 pathways. Finally, our data demonstrated that miR-146b overexpression resulted in a greater suppression of cell migration than STAT3 pathway inhibition alone.These results suggest a complex and heterogeneous role of EGFR in ovarian cancer. Combined blockade of EGFR and IL-6-STAT3 pathways by miR-146b might be a strategy for improving the clinical benefit of targeting the EGFR pathway in ovarian cancer patients in the future. [ABSTRACT FROM AUTHOR]

This study designed to investigate the potential role of lncRNA NEAT1/miR-146b in infantile pneumonia. In this study, 58 children with pneumonia and 58 healthy children collected for routine examination from December 2016 to January 2019. The lncRNA NEAT1 and miR-146b expression levels were detected by qPCR in both groups. The pneumonia model was established by inducing human embryonic lung fibroblasts HFL1 with LPS, and then transfected with lncRNA NEAT1 inhibition and miR-146b over-expression vector to observe the effect on cell viability and apoptosis after induction. Starbase predicted the binding site between lncRNA NEAT1 and miR-146b, and the targeted relationship between them was detected by dual luciferase reporter gene. The relative expression of lncRNA NEAT1 in serum of infantile pneumonia was up-regulated. Knocking down lncRNA NEAT1 promotes cell growth and reduces apoptosis in LPS-induced HFL1 cells. Results showed that the fluorescence activity of lncRNA NEAT1 obviously reduced when combined with miR-146b. In conclusion, the relative expression of miR-146b in serum of infantile pneumonia decreased, and over-expressing it could promote LPS-induced cell viability and reduce apoptosis. Taken together, this study demonstrated that the lncRNA NEAT1 regulates infantile pneumonia by sponging miR-146b. [ABSTRACT FROM AUTHOR]

BACKGROUND: More and more studies have confirmed that miR-146b plays an important role in the nervous system, which provides a new therapeutic direction for treatment of nervous system diseases. OBJECTIVE: To construct rat miR-146b overexpression lentiviral vector and observe the effect of miR-146b overexpression on the proliferation of primary hippocampal neural stem cells. METHODS: (1) PCR extended to obtain the full-length sequence of rat miR-146b, which was ligated into the lentiviral vector pLVX-Luciferase-Puro. After double digestion and DNA sequencing identification, lentivirus was packed in 293T cells and virus supernatant was collected. (2) Primary hippocampal neural stem cells of newborn SD rats were isolated and cultured, and pLVX-Luciferase-rno-miR-146b-Puro lentivirus was transfected into hippocampal neural stem cells. After 24 and 48 hours of transfection, RT-PCR was applied to detect the expression level of miR-146b, while the expression of Nestin was observed by immunofluorescence staining. MTS method and Edu method were used to measure cell proliferation. After 48 hours of transfection, cell cycle was detected by flow cytometry. RESULTS AND CONCLUSION: (1) The results of sequencing identification showed that rat miR-146b was successfully cloned into pLVX-Luciferase-Puro overexpression vector. (2) After 24 and 48 hours of transfection, expression level of miR-146b in hippocampal neural stem cells increased significantly. (3) The number of Nestin immunofluorescence-positive cells was significantly reduced after miR-146 overexpression, and the proliferation ratio was decreased, and the cell cycle mainly remained in G1 phase. (4) The results confirm that the rat miR-146b overexpression lentivirus vector was successfully packaged and it could inhibit the proliferation of primary hippocampal neural stem cells after transfection, but the mechanism of its regulation of target genes needs further study. [ABSTRACT FROM AUTHOR]

aimed to evaluate the efficacy of berberine for improving hepatic insulin resistance and the possible molecular mechanisms involved. Methods: In vitro, HepG2 cells were induced with palmitic acid, and glycogen synthesis was examined. In vivo, a high-fat diet (HFD)-fed mouse model was established, and metabolic parameters were assessed. The expressions of miR-146b and sirtuin 1 (SIRT1) in liver were also examined. The relationship between miR-146b and SIRT1 was examined by the dual-luciferase reporter gene assay. Results: Serum biochemical parameters, such as glucose and HOMA-IR index, were increased in HFD mice; miR-146b and SIRT1 were abnormally expressed in HFD mice and palmitic acid-treated HepG2 cells. Interestingly, berberine reduced body weight and caused a significant improvement in glucose tolerance and HOMA-IR index without altering food intake in mice. Overexpression of miR-146b abolished the protective effect of berberine on palmitic acid-induced impaired glycogen synthesis in HepG2 cells. Luciferase assay showed that miR-146b directly targeted SIRT1. Conclusion: The present findings suggest that berberine could attenuate hepatic insulin resistance through the miR-146b/SIRT1 pathway, which may represent a potential therapeutic target for the prevention and treatment of metabolic diseases, particularly diabetes. [ABSTRACT FROM AUTHOR]

Background: Evidences indicate that exosomes-mediated delivery of microRNAs (miRNAs or miRs) is involved in the neurogenesis of stroke. This study was to investigate the role of exosomal miRNAs in non-drug therapy of electro-acupuncture (EA) regulating endogenous neural stem cells for stroke recovery.Methods: The model of focal cerebral ischemia and reperfusion in rats were established by middle cerebral artery occlusion (MCAO) and treated by EA. The exosomes were extracted from peri-ischemic striatum and identified by exosomal biomarkers, and detected differentially expressed miRNAs with microarray chip. Primary stem cells were cultured, and oxygen–glucose deprivation and reperfusion (OGD/R) was used to mimic vitro ischemic injury.Results: The levels of exosomal biomarkers TSG101 and CD81 were increased in peri-ischemic striatum after EA treatment, and we revealed 25 differentially expressed miRNAs in isolated exosomes, of which miR-146b was selected for further analysis, and demonstrated that EA increased miR-146b expression and its inhibitors could block the effects. Subsequently, we confirmed that EA upregulated miR-146b expression to promote neural stem cells differentiation into neurons in peri-ischemic striatum. In vitro, it was verified that OGD/R hindered neural stem cells differentiation, and miR-146b inhibitors furtherly suppressed its differentiation, simultaneously NeuroD1 was involved in neural stem cells differentiation into neurons. Moreover, in vivo we found EA promoted NeuroD1-mediated neural stem cells differentiation via miR-146b. In addition, EA also could improve neurological deficits through miR-146b after ischemic stroke.Conclusion: EA promotes the differentiation of endogenous neural stem cells via exosomal miR-146b to improve neurological injury after ischemic stroke.

Abstract Background DNA methylation in miRNA genes has been reported as a mechanism that may cause dysregulation of mature miRNAs and consequently impact the gene expression. This mechanism is largely unstudied in papillary thyroid carcinomas (PTC). Methods To identify differentially methylated miRNA-encoding genes, we performed global methylation analysis (Illumina 450 K), integrative analysis (TCGA database), data confirmation (pyrosequencing and RT-qPCR), and functional assays. Results Methylation analysis revealed 27 differentially methylated miRNA genes. The integrative analyses pointed out miR-21 and miR-146b as potentially regulated by methylation (hypomethylation and increased expression). DNA methylation and expression patterns of miR-21 and miR-146b were confirmed as altered, as well as seven of 452 mRNAs targets were down-expressed. The combined methylation and expression levels of miR-21 and miR-146b showed potential to discriminate malignant from benign lesions (91–96% sensitivity and 96–97% specificity). An increased expression of miR-146b due to methylation loss was detected in the TPC1 cell line. The miRNA mimic transfection highlighted putative target mRNAs. Conclusions The increased expression of miR-21 and miR-146b due to loss of DNA methylation in PTC resulted in the disruption of the transcription machinery and biological pathways. These miRNAs are potential diagnostic biomarkers, and these findings provide support for future development of targeted therapies.

Background/Aims: Resveratrol (RSV) has been reported as a promising oral supplementation for osteoarthritis treatment, while the mechanism of its action is still unclear. The specific aim of this study is to decode one of the mechanisms by which RSV protects chondrocyte. Methods: Mouse chondrogenic cell line ATDC5 was treated with 30 µM RSV for 24 h, and 10 µg/ml LPS for 12 h, after which cell viability, apoptosis, and the release of pro-inflammatory cytokines were assessed. The expression of miR-146b in ATDC5 cells was silenced by the specific inhibitor transfection, and then cell viability, apoptosis and inflammation were re-assessed. Results: The IC50 value of LPS in ATDC5 cells was about 10.27 µg/ml. LPS with a dosage of 10 µg/ml repressed cell viability, induced apoptosis, and increased the release of IL-1β, IL-6 and TNF-α. RSV pre-treatment (30 µM) significantly alleviated LPS-induced apoptosis and inflammation. More importantly, miR-146b was up-regulated by RSV, and the protective functions of RSV on ATDC5 cells were attenuated by miR-146b silence. Further, NF-κB and p38MAPK pathways were activated by LPS, and were deactivated by RSV. Besides, RSV-induced the deactivation of NF-κB and p38MAPK pathways was reversed by miR-146b silence. Conclusions: Our findings suggest that RSV protects ATDC5 cells from LPS-induced inflammatory and apoptotic injury via up-regulation of miR-146b and thereby deactivation of NF-κB and p38MAPK pathways.

High mobility group box 1 (HMGB1) is essential for the pathogenesis of liver injury and liver fibrosis. We previously revealed that miR-146b promotes hepatic stellate cells (HSCs) activation and proliferation. Nevertheless, the potential mechanisms are still unknown. Herein, HMGB1 increased HSCs proliferation and COL1A1 and α-SMA protein levels. However, the knockdown of miR-146b inhibited HSCs proliferation and COL1A1 and α-SMA protein levels induced via HMGB1 treatment. miR-146b was upregulated by HMGB1 and miR-146b targeted hepatocyte nuclear factor 1A (HNF1A) 3′-untranslated region (3′UTR) to modulate its expression negatively. Further, we confirmed that HMGB1 might elicit miR-146b expression via p65 within HSCs. Knockdown or block of HMGB1 relieved the CCl4-induced liver fibrosis. In fibrotic liver tissues, miR-146b expression was positively correlated with p65 mRNA, but HNF1A mRNA was inversely correlated with p65, and miR-146b expression. In summary, our findings suggest that HMGB1/p65/miR-146b/HNF1A signaling exerts a crucial effect on liver fibrogenesis via the regulation of HSC function. [ABSTRACT FROM AUTHOR]

Background: Evidences indicate that exosomes-mediated delivery of microRNAs (miRNAs or miRs) is involved in the neurogenesis of stroke. This study was to investigate the role of exosomal miRNAs in non-drug therapy of electro-acupuncture (EA) regulating endogenous neural stem cells for stroke recovery. Methods: The model of focal cerebral ischemia and reperfusion in rats were established by middle cerebral artery occlusion (MCAO) and treated by EA. The exosomes were extracted from peri-ischemic striatum and identified by exosomal biomarkers, and detected differentially expressed miRNAs with microarray chip. Primary stem cells were cultured, and oxygen–glucose deprivation and reperfusion (OGD/R) was used to mimic vitro ischemic injury. Results: The levels of exosomal biomarkers TSG101 and CD81 were increased in peri-ischemic striatum after EA treatment, and we revealed 25 differentially expressed miRNAs in isolated exosomes, of which miR-146b was selected for further analysis, and demonstrated that EA increased miR-146b expression and its inhibitors could block the effects. Subsequently, we confirmed that EA upregulated miR-146b expression to promote neural stem cells differentiation into neurons in peri-ischemic striatum. In vitro , it was verified that OGD/R hindered neural stem cells differentiation, and miR-146b inhibitors furtherly suppressed its differentiation, simultaneously NeuroD1 was involved in neural stem cells differentiation into neurons. Moreover, in vivo we found EA promoted NeuroD1-mediated neural stem cells differentiation via miR-146b. In addition, EA also could improve neurological deficits through miR-146b after ischemic stroke. Conclusion: EA promotes the differentiation of endogenous neural stem cells via exosomal miR-146b to improve neurological injury after ischemic stroke. [ABSTRACT FROM AUTHOR]

Objective: This study aimed to investigate the potential of microRNA (miR)-146a and miR-146b for predicting restenosis and rapid angiographic stenotic progression (RASP) risk in coronary heart disease (CHD) patients who underwent percutaneous coronary intervention (PCI) with drug-eluting stent (DES) implantation. Methods: In total, 255 CHD patients who underwent PCI with DES were enrolled, and their baseline, procedural, and post procedure characteristics were recorded. Plasma samples were obtained before PCI treatment to detect the miR-146a and miR-146b expression by reverse transcription quantitative polymerase chain reaction. Besides, restenosis and RASP occurrences were assessed based on coronary angiograms at 12 months after the surgery. Results: The occurrence rates of restenosis and RASP were 9.0% and 32.9% respectively in CHD patients who underwent PCI with DES. Furthermore, miR-146a and miR-146b expressions were elevated in CHD patients with restenosis compared with CHD patients without restenosis. Subsequent receiver operating characteristic (ROC) curve analysis showed that miR-146a (area under the curve (AUC), 0.674; 95% CI, 0.567-0.781) and miR-146b (AUC, 0.801; 95% CI, 0.729-0.875) could predict increased restenosis risk, among which miR-146b numerically exhibited a better predictive value for higher restenosis risk. Besides, miR-146a and miR-146b expressions were raised in CHD patients with RASP compared with CHD patients without RASP. Followed ROC curve analysis illuminated that miR-146a (AUC, 0.772; 95% CI, 0.714-0.829) and miR-146b (AUC, 0.706; 95% CI, 0.644-0.769) presented similar values in predicting elevated RASP risk. Conclusion: miR-146a and miR-146b predict increased restenosis and RASP risk in CHD patients who underwent PCI with DES. [ABSTRACT FROM AUTHOR]

Background/Aims: Stroke is a major cerebrovascular disease threatening human health and life with high morbidity, disability and mortality. It is aimed to find effective biomarkers for the early diagnosis on stroke. Methods: The expressions of 17 previously reported stroke-associated miRNAs were measured using quantitative RT-PCR and the expressions of plasma high-sensitivity C reactive protein (hs-CRP) and serum interleukin 6 (IL-6), the pro-inflammation markers in brain injury, were examined using enzyme-linked immunosorbent assay in 128 acute ischemic stroke (AIS) patients and control group. Results: Serum miR-146b expression was significantly increased within 24 hours after stroke onset in patients compared with control group. In addition, the upregulation of serum miR-146b was strong positively correlated with plasma hs-CRP, infarct volume and National Institutes of Health Stroke Scale (NIHSS) score, and moderate positively correlated with serum IL-6 of patients. Importantly, the combination of plasma hs-CRP and serum miR-146b gained a better sensitivity/specificity for prediction of AIS (AUC from 0.782 to 0.863). Conclusion: Our preliminary findings suggested that upregulated serum miR-146b in acute ischemic stroke might be a potential biomarker for AIS evaluation.

Background/Aims: Emerging studies on brain-derived neurotrophic factor (BDNF) have shown that might be novel biomarkers and therapeutic targets for cancer. We explore the role of BDNF in the tumorigenesis of bladder cancer and the underlying molecular mechanism. Methods: 368 patients with diagnosed bladder cancer and 352 healthy controls were enrolled to evaluate the association of BDNF and the miR-146b. Bioinformatics algorithm analysis and luciferase assay were performed to identify the target genes of miR-146b. Real-time PCR and western-blot were carried out to validate the relationship between miR-146b and CRK. MTT assay and FACS were used to evaluated the proliferation and apoptosis of cancer cells. MVs were isolated and transfect into the culture cells to confirm the above observation. Results: The clinical study shows that BDNF Met/Met was significantly associated with the risk of bladder cancer. In addition, comparing with Val/Val and Val/Met, Met/Met has lower miR-146b level. Luciferase assay shows that BDNF Val/Val is apparently enhanced miR-146b promoter-luciferase, but not BDNF Met/Met. Based on luciferase assay, CRK is a direct target gene of miR-146b. MiR-146b mimics significantly inhibited the expression of CRK and activation of AKT level. The expression of CRK and the activation of AKT (p-AKT) were significantly inhibited by MV-BDNF Val/Val-miR-146b or MV-BDNF Val/Met-miR-146b, but not MV-BDNF Met/Met-miR-146b. MV-BDNF Val/Val-miR-146b or Val/Met-miR-146b obviously inhibited cell proliferation, which eliminated by CRK. Meanwhile, with MV-BDNF Met/Met-miR-146b or Met/Met-miR-146b+CRK did not affect the proliferation. MV-BDNF Val/Val-miR-146b or Val/Met-miR-146b enhanced cell apoptosis, which could be eliminated by CRK. Meanwhile, MV-BDNF Met/Met-miR-146b or Met/Met-miR-146b+CRK did not promote apoptosis. Conclusion: BDNF VAL66MET polymorphism is associated with miR-146b and its target gene CRK. MiR-146b and CRK mediated BDNF VAL66MET polymorphism associated proliferation and apoptosis via activation of AKT. Thus, BDNF Val66Met is associated with the risk of bladder cancer, and the BDNF variant could be used a biomarker for the diagnosis of bladder cancer.

Abstract Background Weaning stress affects the small intestine of piglets. MiR-146b is differentially expressed in suckling and weaned piglets. In this study, we evaluated the effects of miR-146b on cell viability, proliferation, and apoptosis in IPEC-J2 cells. Results Transfection with miR-146b mimics successfully increased miR-146b levels by 1000× (P

Background/aims: The purpose of this study is to explore the inhibition or activation effects of microRNA-146 B on the expression of PTP1B in gastric cancer cells.Methods: The expressions of PTP1B and miR-146b in gastric cancer were detected by RT-qPCR. The effects of miR-146b on cell apoptosis and proliferation of gastric cancer were detected. The methods used in the detection process included Annexin V/PI dying method, colony formation assay, and MTT assay. The downstream target gene miR-146b was predicted and screened by bioinformatics and luciferase reporter assay. The mRNA and protein expressions of the target gene PTP1B miR-146b were determined using RT-qPCR and western blot. The expression of miR-146 B in mice was detected by the cells transfected with microRNA-146 B in vivo.Results: Compared with normal tissues, PTP1B was higher and miR-146b was lower in cancer cells. Over-expression of miR-146b can inhibit cell viability and increase the apoptosis rate. According to the luciferase reporter assay, PTP1B was the downstream target gene of miR-146b. The re-introduction of PTP1B reversed the growth inhibition and apoptosis of gastric cancer cells induced by miR-146b. From the mouse xenograft model, the over-expression of miR-146b inhibited the tumor growth and reduced the expression level of PTP1B.Conclusion: miR-146b directly inhibits the expression of PTP1B and suppressed the growth and development of gastric cancer. [ABSTRACT FROM AUTHOR]

Background: Pneumonia is a common respiratory disease worldwide that can be prevented and treated. However, it is considered to be the leading cause of children death. The present study was aimed to explore the functional role and molecular mechanism of miR-146b in the inflammation injury in pediatric pneumonia. Materials and methods: The lipopolysaccharide (LPS)-induced pulmonary injury cell model was established in WI-38 human lung fibroblasts cells. QRT-PCR and Western blot was applied to detect miR-146b and MyD88 expression. ELISA assay was used to analyze the production of pro-inflammatory factors. Cell viability was evaluated by CCK-8 assay. The apoptosis proteins and the downstream genes of NF-jB pathway were detected by Western blot. Results: we displayed that miR-146b was down-regulated, whereas MyD88 was up-regulated in the serum of children patients with pneumonia and in WI-38 cells treated with LPS. Moreover, re-expression of miR-146b suppressed the production of inflammatory factors in the serum of pneumonia patients and WI-38 cells treated with LPS. In addition, elevating miR-146b expression increased WI-38 cell viability and reduced cell apoptosis. More importantly, bioinformatics analysis revealed that MyD88 was a target of miR-146b and could overturn the protective effect of miR-146b on the inflammation injury in LPS-injured WI-38 cells. Furthermore, miR-146b over-expression inhibited the activation of NF-jB signaling pathway by suppressing MyD88. Conclusion: miR-146b attenuated the inflammation injury in pediatric pneumonia through inhibiting MyD88/NF-jB signaling pathway. These preliminarily findings further deepened our understanding of this mechanism and identified new potential therapeutic targets for pediatric pneumonia. [ABSTRACT FROM AUTHOR]

Background and Aims Macrophages are a promising therapeutic target for intestinal mucosal repair. MiR-146b appears to control macrophage activation and cell proliferation. Methods By loading miR-146b mimic on mannose-modified trimethyl chitosan [MTC]-conjugated nanoparticles [NPs] [MTC-miR146b], a molecular targeted immunotherapeutic approach was developed to selectively target intestinal macrophages for mucosal regeneration and tumourigenesis in mouse models. Results We first confirmed that miR-146b expression was significantly enhanced during mucosal regeneration in a murine colitis model. Moreover， after mucosal damage, MTC-miR146b mimic-treated wild-type mice had dramatically restored body weight and mucosal barrier function compared with MTC-NC treated mice. Strikingly, MTC-miR146b mimic oral administration protected miR-146b-deficient mice from dextran sodium sulphate [DSS] injury and the colitis-associated cancer process. Mechanistically, miR-146b strongly inhibited M1 macrophage activation by suppressing the Toll-like receptor 4 [TLR4] signalling pathway, resulting in the repression of the induction of pro-inflammatory cytokines including TNF-α, IL-6, and IL-1β. More importantly, miR-146b overexpression in bone marrow-derived macrophages [BMDMs] in M1 differentiation conditions induced a phenotype similar to M2 macrophages and improved the proliferation of co-cultured colonic epithelial cells via STAT3-dependent IL-10 production. Conclusions MTC-miR146b should be regarded as an effective candidate for oral delivery and could improve the efficacy of immunotherapies for ulcerative colitis and colitis-associated cancer. [ABSTRACT FROM AUTHOR]

Background/Aims: The primary objective of this study is to evaluate the diagnosis effect of serum protein factors and microRNAs for children suffering from viral myocarditis (VMC). Methods: The expression levels of serum cardiac troponin I (cTnI), interleukin-18 (IL-18) and tumor necrosis factor-α (TNF-α) in both VMC and control groups were examined by using the Elisa Kit. The expression levels of miR-1 and miR-146b were measured through RT-PCR. Subsequently, the Receiver Operating Characteristic (ROC) curves were drawn based on the diagnostic results of VMC. Moreover, the Spearman correlation analysis was carried out to unveil the association between the indicator expression levels and the ultrasonic cardiogram results, including the left ventricular fractional shortening (FS) and left ventricular ejection fraction (EF). Results: It is found that the expression levels between the VMC and control group portrait significant differences with respect to cTnI, IL-18, TNF-α, miR-1 and mIR-146b (P Conclusions: The expression levels of the TNF-α, IL-18 and cTnI and the expression levels of the miR-1 and miR-146b could be used to predict VMC among children and this approach may reinforce the diagnosis of VMC in clinical practices.

Background. We previously identified miR-146b as being up-regulated during the development of hepatic fibrosis using deep sequencing technology and gene expression analysis. However, the roles and related mechanisms of miR-146b in hepatic stellate cells (HSCs), which are involved in fibrogenesis and fibrosis, have not been elucidated.Results. We report that miR-146b expression was increased in TGF-β1-treated HSCs. TGF-β1 enhanced a-SMA and COL1A1 protein expression in HSCs and stimulated proliferation of these cells compared with cells transfected with inhibitor NC. Conversely, miR-146b knock-down decreased α-SMA and COL1A1 expression and inhibited HSC proliferation. In addition, we found that miR-146b specifically regulated the translation of Krüppel-like factor 4 (KLF4) by targeting its 3’ untranslated region. Forced expression of KLF4 inhibited TGF-β1-induced enhancement of α-SMA and COL1A1 expression in HSCs, as well as proliferation of these cells. Moreover, miR-146b expression was negatively associated with KLF4 expression but positively associated with expression of α-SMA and COL1A1 during hepatic fibrosis.Conclusions. Our findings demonstrate the participation of miR-146b as a novel upstream effector of HSC activation via direct targeting of KLF4. Thus, targeted transfer of miR-146b into HSCs could be a useful strategy for the treatment of hepatic fibrosis.

Viral myocarditis (VMC) is a common cardiac disease, however, there still lacks an effective therapeutic strategy for VMC. Astragalus mongholicus (Fisch.) Bge (AB), a Chinese herb with some functional metabolites, may have some pharmacological effects on VMC. AB ingredients were measured by a full-scan LCQ mass spectrum. We aimed to explore the effects of AB on the VMC children by investigating peripheral Treg cell homeostasis. A total of 68 VMC children were random and evenly assigned into an AG group (received 10-mL AB oral liquid daily), and a CG group (received placebo daily). Peripheral blood mononuclear cells (PBMC) were obtained from peripheral blood and Treg cells were isolated. The levels of miR-146b, miR-155, Treg immunity activity and myocarditis biomarkers were measured in Treg cells. There were four main components (sucrose, calycosin, Astragaloside IV and calycosin-7-glucoside) in AB. The cases sinus tachycardia, frequent premature ventricular contractions, and supraventricular tachycardia were significantly reduced in the AG group (P < 0.05). Meanwhile, the myocardial enzymes and cardiac function indexes were improved in the AG group when compared with the CG group (P < 0.05). The time of electrocardiogram recovery, symptom duration and hospital stay was shorter in the AG group than in the CG group (P < 0.05). The levels of miR-146b and miR-155 were higher in the CG group than in the AG group (P < 0.05). The levels of ROR-γt (retinoic acid receptor-related orphan nuclear receptor gamma), FoxP3 (forkhead transcription factor), IL-10 (interleukin-11) and TGF-β (transforming growth factor beta) were lower in the CG group than in the AG group (P < 0.05). In contrast, the levels of IL-17, IL-21, CK-MB (creatine kinase-MB), cTnI (cardiac troponin I), GrB (granzyme B), sFasL (soluble fas ligand) and caspase-3 were higher in the CG group than in the AG group (P < 0.05). Furthermore, the levels of ROR-γt, FoxP3, IL-10, and TGF-β were positively, whereas the levels of IL-17, IL-21, CK-MB, cTnI, GrB, sFasL and caspase-3 were negatively, associated with the levels of miR-146b and miR-155 (P < 0.05). AB treatment improved cardiac functions, peripheral Treg cell immunity imbalance in the children with VMC by reducing the levels of miR-146b and miR-155.

Objective To explore the expression and clinical significance of plasma miR-1 and miR-146b in patients with acute ST-segment elevation myocardial infarction(STEMI). Methods Seventy-nine cases of STEMI patients(observation group) and 59 cases of patients with non-STEMI chest pain(control group) were selected. There was no significant difference in gender, age,history of hypertension,history of diabetes,smoking history, bMI,systolic blood pressure, diastolic blood pressure or TC levels between the two groups(all P > 0. 05). The levels of creatine kinase isoenzyme(CKMB) and cardiac troponin I(cTnI) in the two groups were measured, and the expression of plasma miR-1 and miR-146b in the two groups at the time of on admission(T0) and at 12 h(T1),24 h(T2),48 h(T3), and 7 d(T4) after admission was detected. Results The plasma levels of CK-MB and cTnI in the observation group were higher than those in the control group(all P < 0. 05). The expression levels of plasma miR-1 and miR-14b in patients with STEMI at T0, t1, t2 and T3 was significantly higher than those of the control group(all P < 0. 05). The levels of plasma miR-1 and miR-146b were the highest at T1, and there was a statistically significant difference in comparison with other time points(P < 0. 05).The expression levels of plasma miR-1 and miR-146 b in STEMI patients was positively correlated with CK-MB, CTnI and coronary angiography score(all P < 0. 05). Conclusion The elevated level of miR-1 and miR-146 b in STEMI patients is expected to become a new target for STEMI diagnosis and treatment. [ABSTRACT FROM AUTHOR]

Background/Aims: Resveratrol (RSV) has been reported as a promising oral supplementation for osteoarthritis treatment, while the mechanism of its action is still unclear. The specific aim of this study is to decode one of the mechanisms by which RSV protects chondrocyte. Methods: Mouse chondrogenic cell line ATDC5 was treated with 30 µM RSV for 24 h, and 10 µg/ml LPS for 12 h, after which cell viability, apoptosis, and the release of pro-inflammatory cytokines were assessed. The expression of miR-146b in ATDC5 cells was silenced by the specific inhibitor transfection, and then cell viability, apoptosis and inflammation were re-assessed. Results: The IC50 value of LPS in ATDC5 cells was about 10.27 µg/ml. LPS with a dosage of 10 µg/ml repressed cell viability, induced apoptosis, and increased the release of IL-1β, IL-6 and TNF-α. RSV pre-treatment (30 µM) significantly alleviated LPS-induced apoptosis and inflammation. More importantly, miR-146b was up-regulated by RSV, and the protective functions of RSV on ATDC5 cells were attenuated by miR-146b silence. Further, NF-κB and p38MAPK pathways were activated by LPS, and were deactivated by RSV. Besides, RSV-induced the deactivation of NF-κB and p38MAPK pathways was reversed by miR-146b silence. Conclusions: Our findings suggest that RSV protects ATDC5 cells from LPS-induced inflammatory and apoptotic injury via up-regulation of miR-146b and thereby deactivation of NF-κB and p38MAPK pathways. [ABSTRACT FROM AUTHOR]

BACKGROUND: To facilitate the neuronal differentiation of induced pluripotent stem cells (iPSCs) is of great benefit for the repair of nerve injury and nerve regeneration. OBJECTIVE: To investigate the expression of miR-146b in iPSCs differentiation, and its effects on the neural differentiation of iPSCs. METHODS: After 7-day neural induction of mouse iPSCs, the expressions of miR-146b and pluripotent stem cell markers were detected by real-time PCR. Then, we generated miR-146b overexpressing iPSCs followed by the neural induction. The expression of stem cell and neuroectoderm markers were examined by cell immunofluorescence and real-time PCR respectively. Meanwhile, the expression of Tuj-1, a nerve cell marker, was also detected. RESULTS AND CONCLUSION: Compared with the normal iPSCs, the expressions of pluripotent stem cell markers Nanog, Oct4 and Sox2 were significantly down-regulated in iPSCs undergoing 7-day neural induction, while the level of miR-146b was increased obviously. In miR-146b-overexpressing iPSCs, we found that the expression of Oct4 was substantially decreased, while there were no statistical changes in the expression of Nanog and Sox2. Meanwhile, during the neural differentiation, miR-146b overexpression significantly down-regulated the expression of Nanog, Oct4 and Sox2, and up-regulated the expression of neuroectoderm markers, Nestin, Sox1 and Pax6. After 18 days of neural induction, the miR-146b overexpression significantly increased the number of Tuj-1-positive cells. Taken together, miR-146b plays a vital role in facilitating the differentiation of iPSCs into neuron-like cells. [ABSTRACT FROM AUTHOR]

A previous study has indicated that Krüppel-like factor 7 (KLF7), a transcription factor that stimulates Schwann cell (SC) proliferation and axonal regeneration after peripheral nerve injury, is a promising therapeutic transcription factor in nerve injury. We aimed to identify whether inhibition of microRNA-146b (miR-146b) affected SC proliferation, migration, and myelinated axon regeneration following sciatic nerve injury by regulating its direct target KLF7. SCs were transfected with miRNA lentivirus, miRNA inhibitor lentivirus, or KLF7 siRNA lentivirus in vitro. The expression of miR146b and KLF7, as well as SC proliferation and migration, were subsequently evaluated. In vivo, an acellular nerve allograft (ANA) followed by injection of GFP control vector or a lentiviral vector encoding an miR-146b inhibitor was used to assess the repair potential in a model of sciatic nerve gap. miR-146b directly targeted KLF7 by binding to the 3′-UTR, suppressing KLF7. Up-regulation of miR-146b and KLF7 knockdown significantly reduced the proliferation and migration of SCs, whereas silencing miR-146b resulted in increased proliferation and migration. KLF7 protein was localized in SCs in which miR-146b was expressed in vivo. Similarly, 4 weeks after the ANA, anti-miR-146b increased KLF7 and its target gene nerve growth factor cascade, promoting axonal outgrowth. Closer analysis revealed improved nerve conduction and sciatic function index score, and enhanced expression of neurofilaments, P0 (anti-peripheral myelin), and myelinated axon regeneration. Our findings provide new insight into the regulation of KLF7 by miR-146b during peripheral nerve regeneration and suggest a potential therapeutic strategy for peripheral nerve injury. [ABSTRACT FROM AUTHOR]

Non-alcoholic fatty liver disease (NAFLD) is one of the most common chronic liver diseases worldwide, and precision therapeutic will be a benefit for the NAFLD regression. In this study, we observed low microRNA 146 b (miR-146 b) expression in NAFLD mice model induced by methionine-choline-deficient diet (MCD) compared with control group. Furthermore, miR-146b−/− mice induced MCD exhibited severe liver steatosis and hepatitis. A bio-distribution study showed that novel Lactosylated PDMAEMA nanoparticles effectively targeted hepatocytes Lac-PDMAEMA. We coupled miR-146b mimic with Lac-PDMAEMA and then were administrated to NAFLD mice model, which could obviously alleviate the hepatic steatosis. Lac-PDMAEMA effectively delivered miR-146b mimic to hepatocytes with a ∼8-fold upregulation of miR-146b mimic targeting MyD88 and IRAK1, and in turn suppressed the expression of PPARγ. Meanwhile, TNF-α and IL-6 mRNA levels were decreased after administration of Lac-PDMAEMA/miR-146b mimic. So, we made a conclusion that targeted delivering miR-146b mimic to the hepatocytes by, coupling Lac-PDMAEMA nanoparticles could effectively alleviate the hepatic steatosis in NAFLD mice, which maybe bring a new and effective way to intervene and therapy the NAFLD. [ABSTRACT FROM AUTHOR]

Adipose tissue plays an important role in energy metabolism. Adipose dysfunction is closely related to obesity and type II diabetes. Glucose uptake is the key step for fat synthesis in adipocyte. miRNAs have been proven to play a crucial role in adipocyte differentiation, adipogenesis and glucose homeostasis. In this paper, we firstly reported that miR-146b decreased glucose consumption by up-regulating miR-146b in a porcine primary adipocyte model, while the inhibitor of endogenous miR-146b rescued the reduction. Then, miR-146b was predicated to target IRS1 by bioinformatics analysis, and a dual-luciferase reporter assay validated this predication. Western blot analyses indicated both IRS1 and glucose transporter type 4 (GLUT4) were down-regulated by miR-146b overexpression. Our study demonstrated that miR-146b regulated glucose homeostasis in porcine primary pre-adipocyte by targeting IRS1, and provided new understandings on regulations of lipogenesis by miRNAs. [ABSTRACT FROM AUTHOR]

This study evaluated the gene expression of the pro-inflammatory cytokines, interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-α) in canine mammary tumors (CMTs), and correlated them with gene expression of miRNAs expected to regulate the secretion of pro-inflammatory cytokines within the tumor microenvironment (TME). Furthermore, gene expression of cytokines and miRNAs involved in tumor cell proliferation and invasion (i.e. miR-21; miR-124; miR-145) were correlated with tumor proliferation index (Ki67 index) to determine the prognostic value in CMTs. Twenty-six canine mammary samples were used, including 22 CMTs and 4 control samples. MiR-21, IL-6 and TNF-α were upregulated in mammary carcinomas compared with controls (p < 0.05). MiR-146b was downregulated in CMTs compared with control cases (p < 0.05). IL-6 expression showed a significant positive correlation with miR-21 and a negative correlation with miR-146b; while, TNF-α gene expression was positively correlated with miR-21 and miR-145 in mammary carcinomas. In carcinomas, the Ki67 index correlated positively with gene expression of IL-6 and miR-21 and negatively correlated with miR-145 and miR-146b. Specifically, gene expression of IL-6 and miR-21 was positively correlated with ki67 index >33.3%, whereas, expression of miR-145 and miR-146b was negatively correlated with ki67 index <33.3%. Results reinforce the concept of interaction between tumor cells and inflammatory cells within the TME, with a central role of IL-6 and TNF-α. Since the upregulation of miR-21 reflects the gene overexpression of interleukins and the high proliferation index of tumor cells, this miRNA may be considered a biomarker with prognostic value in CMTs., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2023 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Background/Aims: The primary objective of this study is to evaluate the diagnosis effect of serum protein factors and microRNAs for children suffering from viral myocarditis (VMC). Methods: The expression levels of serum cardiac troponin I (cTnI), interleukin-18 (IL-18) and tumor necrosis factor-α (TNF-α) in both VMC and control groups were examined by using the Elisa Kit. The expression levels of miR-1 and miR-146b were measured through RT-PCR. Subsequently, the Receiver Operating Characteristic (ROC) curves were drawn based on the diagnostic results of VMC. Moreover, the Spearman correlation analysis was carried out to unveil the association between the indicator expression levels and the ultrasonic cardiogram results, including the left ventricular fractional shortening (FS) and left ventricular ejection fraction (EF). Results: It is found that the expression levels between the VMC and control group portrait significant differences with respect to cTnI, IL-18, TNF-α, miR-1 and mIR-146b (P < 0.05). The diagnostic thresholds for cTnI, IL-18, TNF-α were 160.00 ng/L, 2.26 ng/L and 1.14ng/L, respectively. The diagnostic thresholds for miR-1 and miR-146b were 0.75 and 1.27, respectively. Results from the Spearman correlation analysis showed that levels of the miR-1 were negatively correlated with FS and EF, while levels of the cTnI, IL-18, TNF-α and miR-146b were positively correlated with FS and EF. Conclusions: The expression levels of the TNF-α, IL-18 and cTnI and the expression levels of the miR-1 and miR-146b could be used to predict VMC among children and this approach may reinforce the diagnosis of VMC in clinical practices. [ABSTRACT FROM AUTHOR]

Objective Toll-like receptor signaling activated by bacterial otitis media pathogens in the middle ear has been shown to play a key role in OM susceptibility, pathogenesis and recovery. Recent studies implicate microRNA 146 (miR-146) in regulation of inflammation via negative feedback of toll-like receptor signaling (TLR) in a wide variety of tissues, however its involvement in otitis media is unknown. Methods Human middle ear epithelial cells were stimulated with proinflammatory cytokines, interleukin 1 beta or tumor necrosis factor alpha, for two to twenty-four hours. Middle ear biopsies were collected from children with otitis media with effusion (n = 20), recurrent otitis media (n = 9), and control subjects undergoing cochlear implantation (n = 10). miR-146a, miR-146b expression was assayed by quantitative PCR (qPCR). Expression of miR-146 targets involved in TLR signaling, IRAK1 and TRAF6, was assayed by qPCR in middle ear biopsies. Middle ear biopsies were cryosectioned and epithelial thickness measured by a certified pathologist. Results Proinflammatory cytokines induced expression of miR-146 in middle ear epithelial cells in vitro . Middle ear miR-146a and miR-146b expression was elevated in otitis media patients relative to control subjects and correlated with middle ear epithelial thickness. A trend towards inverse correlation was observed between miR-146 and TRAF6 expression in the clinical population. Conclusions This report is the first to assess miRNA expression in a clinical population with OM. Findings herein suggest miR-146 may play a role in OM. [ABSTRACT FROM AUTHOR]

Introduction: Micro-RNAs are a new class of regulatory RNAs with length of 18-22 nucleotides involved in the pathogenesis of many human cancers including papillary thyroid cancer (PTC). In general, it acts as an oncogenes or tumor suppressor. This study examined the expression levels of miR-146b in patient and healthy people plasma to find out that if it can be used as an early diagnostic indicator in papillary thyroid cancer. Materials and Methods: in this study, we examined the expression level of miR-146b using q rt-PCR method in plasma of 40 patients, divided into four groups (10 PTC patients on the day before the operation, 10 patients with benign nodules and 10 patients after the Thyroidectomy and 10 healthy subjects as control). All data were normalized with expression level of GAPDH, as an internal control. Findings: expression level of miR-146b in plasma samples of patients was about 5.5 times greater than that in control patients (P <0.05), and generally statistical difference was found in plasma levels for the expression of miR-146b between controls and patients with PTC. However, significant difference was not found in the expression level of miR-146b between control subjects, thyroidectomy and benign patients. Conclusion: This study showed that increased expression of miR-146b and its presence in blood circulation are directly correlated with PTC. The results also indicated the capacity of this technique in distinguishing benign, thyroidectomy, PTC, and normal groups. Therefore, this method can be introduced as a non-invasive method, versus previous methods like FNA, aid in diagnosis to pathologist. [ABSTRACT FROM AUTHOR]

Background: Tumor invasiveness is directly related to the ability of tumor cells to migrate and invade surrounding tissues, usually degrading extracellular matrix. Despite significant progress in the knowledge about migration and invasion, there is much more to elucidate about their regulatory mechanisms, especially in cancer cells. MicroRNAs (miRs) were recently described as important regulators of migration. Differential expression of miRs in cancer is frequently associated with progression, invasion and metastasis. In papillary thyroid carcinoma (PTC), miR-146b-5p is highly expressed and positively correlated to the degree of malignancy.Methods: This study aimed to investigate the role of miR-146b-5p on the migratory and invasive behaviors of thyroid cells, using a non tumor rat thyroid follicular cell line (PCCl3) transfected with the miR-146b-5p genomic region, and two PTC cell lines (TPC-1 and BCPAP, bearing distinct oncogenic backgrounds), which express high levels of miR-146b-5p, after miR-146b inhibition by antagomiR and miR-146b overexpression by mimics-miR. Migration and invasion were studied by time-lapse and transwell assays (with and without Matrigel®). Gelatin degradation assays were also employed, as well as F-actin staining.Results: Migration and invasion of PCCl3 were increased 2-3x after miR-146b-5p overexpression (10X) and large lamellipodia were evident in those cells. After miR-146b-5p inhibition, TPC-1 and BCPAP migration and invasion were significantly reduced, with cells showing several simultaneous processes and low polarity. Gelatin degradation was inhibited in TPC-1 cells after inhibition of miR-146b-5p, but was unaffected in BCPAP cells, which did not degrade gelatin. The inhibition of miR-146b-5p in PCCl3 also inhibited migration and invasion, and additional (exogenous) overexpression of this miR in TPC-1 and BCPAP cells increased migration and invasion, without effects on cell morphology or gelatin degradation. The overexpression of SMAD4 in BCPAP cells, a validated target of miR-146b-5p and key protein in the TGF-β signaling pathway, inhibited migration similarly to the effects observed with the antagomiR 146b-5p.Conclusions: miR-146b-5p positively regulates migration and invasion of thyroid normal and tumor follicular cells (independently from their original mutation, either BRAF or RET/PTC), through a mechanism that involves the actin cytoskeleton but not an increased capacity of matrix degradation. [ABSTRACT FROM AUTHOR]

The major goal of this study was to perform an in depth characterization of the “gene signature” of human FoxP3 + T regulatory cells (Tregs). Highly purified Tregs and T conventional cells (Tconvs) from multiple healthy donors (HD), either freshly explanted or activated in vitro , were analyzed via RNA sequencing (RNA-seq) and gene expression changes validated using the nCounter system. Additionally, we analyzed microRNA (miRNA) expression using TaqMan low-density arrays. Our results confirm previous studies demonstrating selective gene expression of FoxP3 , IKZF2 , and CTLA4 in Tregs. Notably, a number of yet uncharacterized genes ( RTKN2 , LAYN , UTS2 , CSF2RB , TRIB1 , F5 , CECAM4 , CD70 , ENC1 and NKG7 ) were identified and validated as being differentially expressed in human Tregs. We further characterize the functional roles of RTKN2 and LAYN by analyzing their roles in vitro human Treg suppression assays by knocking them down in Tregs and overexpressing them in Tconvs. In order to facilitate a better understanding of the human Treg gene expression signature, we have generated from our results a hypothetical interactome of genes and miRNAs in Tregs and Tconvs. [ABSTRACT FROM AUTHOR]

Studies have shown that the calcium-sensing receptor (CaSR) mediates the antitumorigenic effects of calcium against colorectal cancer (CRC). Expression of the CaSR in colorectal tumors is often reduced. We have reported previously that silencing of CaSR in CRC is caused in part by methylation of CaSR promoter 2 and loss of histone acetylation. We investigated the impact of aberrant microRNA expression on loss of CaSR expression. A microarray study in two Caco-2 subclones (Caco2/AQ and Caco2/15) that have similar genetic background, but different CaSR expression levels (Caco2/AQ expressing more CaSR than Caco2/15), identified 22 differentially expressed microRNAs that potentially target the CaSR. We validated these results by performing gain- and loss-of-function studies with the top candidates: miR-9, miR-27a, miR-135b, and miR-146b. Modulation of miR-135b or miR-146b expression by mimicking or inhibiting their expression regulated CaSR protein levels in two different colon cancer cell lines: Caco2/AQ (moderate endogenous CaSR expression) and HT29 (low endogenous CaSR levels). Inhibition of miR-135b and miR-146b expression led to high CaSR levels and significantly reduced proliferation. In samples of colorectal tumors we observed overexpression of miR-135b and miR-146b, and this correlated inversely with CaSR expression (miR-135b: r = −0.684, p < 0.001 and miR-146b: r = −0.448, p < 0.001), supporting our in vitro findings. We demonstrate that miR-135b and miR-146b target the CaSR and reduce its expression in colorectal tumors, reducing the antiproliferative and prodifferentiating actions of calcium. This provides a new approach for finding means to prevent CaSR loss, developing better treatment strategies for CRC. [ABSTRACT FROM AUTHOR]