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Background： At present, a diagnostic tool with high specificity for impaired endometrial receptivity, which may lead to implantation failure, remains to be developed. We aimed to assess the different endometrial microRNA （miRNA） signatures for impaired endometrial receptivity by microarray analysis. Methods： A total of 12 repeated implantation failure （RIF） patients and I0 infertile patients, who conceived and delivered after one embryo transfer attempt, were recruited as RIF and control groups, respectively. Endometrial specimens from the window of implantation （WOI） were collected from these two groups. MiRNA microarray was conducted on seven and five samples from the RIF and control groups, respectively. Comparative, functional, and network analyses were performed for the microarray results. Quantitative real-time polymerase chain reaction （PCR） was performed on other samples to validate the expression of specific miRNAs. Results： Compared with those in the control group, the expression levels of 105 miRNAs in the RIF group were found to be significantly up- or down-regulated （at least 2-fold） by microarray analysis. The most relevant miRNA functional sets of these dysregulated miRNAs were miR-30 family, human embryonic stern cell regulation, epithelial-mesenchymal transition, and miRNA tumor suppressors by tool for annotations ofmicroRNA analysis. Network regulatory analysis found 176 miRNA-mRNA interactions, and the top 3 core miRNAs were has-miR-4668-5p, has-miR-429, and has-miR-5088. Expression levels of the 18 selected miRNAs in new samples by real-time PCR were found to be regulated with the same trend, as the result ofmicroarray analysis. Conclusions： There is a significant different expression of certain miRNAs in the WOI endometrium for RIF patients. These miRNAs may contribute to impaired endometrial receptivity.

MicroRNAs (miRNAs) are a class of small noncoding RNAs ~22 nt in length that regulate gene expression and play fundamental roles in multiple biological processes, including cell differentiation, proliferation and apoptosis as well as disease processes. The study of miRNA has thus become a rapidly emerging field in life science. The detection of miRNA expression is a very important first step in miRNA exploration. Several methodologies, including cloning, northern blotting, real-time RT-PCR, microRNA arrays and ISH (in situ hybridization), have been developed and applied successfully in miRNA profiling. This review discusses the main existing microRNA detection technologies, while emphasizing microRNA arrays.

miRNA profile deregulation affecting downstream signaling pathways activates endpoints that represent potential biomarkers for prognosis and treatment of tumor patients. In the past 20 years conventional therapy for osteosarcoma (OS) reached a survival plateau, highlighting the need for new therapeutic approaches. In this study, microarray unsupervised and supervised analysis identified, respectively, 100 and 40 differentially expressed miRNAs in OS samples with different grades of malignancy compared to normal bone. When analyzing low-grade and high-grade OS by unsupervised analysis, 12 miRNAs were found to be differentially expressed. Real-time PCR performed on a larger series of OS confirmed a significant lower expression of miR-1, miR-133b and miR-378* in tumors with respect to control, also showing lower mRNA levels in 31 high-grade OS than in 25 low-grade and in metastatic versus non-metastatic patients. We demonstrated that miR-1 and miR133b were downregulated in OS cell lines compared to normal osteoblasts. Secondly, by transfection with miRNA precursor molecules, we demonstrated that the ectopic expression of miR-1 and miR-133b in U2-OS cell lines significantly reduced cell proliferation and MET protein expression and negatively regulated cell invasiveness and motility in a short-term assay. Cell cycle distribution revealed block in G1 and delay of cell cycle progression associated with increased apoptosis in miR-1- and miR-133b-transfected cells, respectively. Our data assessed specific miRNA profiling deregulation in OS clinical samples and suggest that the expression of miR-1 and miR-133b may control cell proliferation and cell cycle through MET protein expression modulation.

Purpose To identify the expression profile of novel microRNAs (miRNAs) in colon cancer and evaluate their clinical applicability. Methods Differences in the expression of serum miRNAs in patients with colon cancer and healthy controls were identified using miRNA microarrays. Differentially expressed miRNAs were verified via real-time polymerase chain reaction (PCR) using sera from 50 patients with colon cancer and 44 healthy controls. These miRNAs were also verified in a double-blind validation experiment using sera from 30 patients with colon cancer, 30 patients with colonic polyps, and 30 healthy controls. Results Microarray hybridization revealed that 87 miRNAs were differentially expressed between the sera of patients with colon cancer and healthy controls. Among these miRNAs, 39 miRNAs were up-regulated, whereas 48 miRNAs were down-regulated. Verification of the expression of these miRNAs using real-time PCR revealed that the expression levels of miR - 31 , miR - 141 , miR - 224 - 3p , miR - 576 - 5p , and miR - 4669 were significantly different between patients with colon cancer and healthy controls. Using these five miRNAs to construct a miRNA expression profile (or miRNA panel) will facilitate more effective diagnosis of colon cancer. Conclusion Clinical analysis of miR - 31 , miR - 141 , miR - 224 - 3p , miR - 576 - 5p , and miR - 4669 expression in patients with colon cancer may facilitate the diagnosis of colon cancer.

Airway remodeling is a significant pathological change of asthma. This study aimed to detect differentially expressed microRNAs in the serum of asthma patients and airway smooth muscle cells (ASMCs) of asthmatic mice, exploring their role in the airway remodeling of asthma. The differentially expressed microRNAs in the serum of mild and moderate-severe asthma patients compared to healthy subjects were revealed using the \"limma\" package. Gene Ontology (GO) analysis was used to annotate the functions of microRNA target genes. The relative expressions of miR-107 (miR-107-3p in mice sharing the same sequence) in the primary airway smooth muscle cells (ASMCs) of the asthma mice model were tested by RT-qPCR. Cyclin-dependent kinases 6 (Cdk6), a target gene of miR-107, was predicted by algorithms and validated by dual-luciferase reporter assay and Western blot. The roles of miR-107, Cdk6, and protein Retinoblastoma (Rb) in ASMCs were examined by transwell assay and EDU KIT in vitro. The expression of miR-107 was down-regulated in both mild and moderate-severe asthma patients. Intriguingly, the level of miR-107 was also decreased in ASMCs of the asthma mice model. Up-regulating miR-107 suppressed ASMCs' proliferation by targeting Cdk6 and the phosphorylation level of Rb. Increasing the expression of Cdk6 or suppressing Rb activity abrogated the proliferation inhibition effect of ASMCs induced by miR-107. In addition, miR-107 also inhibits ASMC migration by targeting Cdk6. The expression of miR-107 is down-regulated in serums of asthma patients and ASMCs of asthmatic mice. It plays a critical role in regulating the proliferation and migration of ASMCs via targeting Cdk6.

After spinal cord injury, dysregulated miRNAs appear and can participate in inflammatory responses, as well as the inhibition of apoptosis and axon regeneration through multiple pathways. However, the functions of miRNAs in spinal cord ischemia-reperfusion injury progression remain unclear. miRCURY LNATM Arrays were used to analyze miRNA expression profiles of rats after 90 minutes of ischemia followed by reperfusion for 24 and 48 hours. Furthermore, subsequent construction of aberrantly expressed miRNA regulatory patterns involved cell survival, proliferation, and apoptosis. Remarkably, the mitogen-activated protein kinase (MAPK) signaling pathway was the most significantly enriched pathway among 24- and 48-hour groups. Bioinformatics analysis and quantitative reverse transcription polymerase chain reaction confirmed the persistent overexpression of miR-22-3p in both groups. These results suggest that the aberrant miRNA regulatory network is possibly regulated MAPK signaling and continuously affects the physiological and biochemical status of cells, thus participating in the regulation of spinal cord ischemia-reperfusion injury. As such, miR-22-3p may play sustained regulatory roles in spinal cord ischemia-reperfusion injury. All experimental procedures were approved by the Animal Ethics Committee of Jilin University, China [approval No. 2020 (Research) 01].

The identification of antemortem burns and postmortem burns is essential in forensic practice. In this study, microRNA (miRNA) microarray analysis was conducted to identify differentially expressed miRNAs in the skin of an experimental burn model. Microarray analysis revealed 24 differentially expressed miRNAs in antemortem burned mice skin, with 19 miRNAs significantly upregulated and 5 downregulated. Based on the intersection predicted using three databases (Targetscan, microRNA.org, and PITA), 293 potential miRNA targets were identified. These dysregulated miRNAs and their predicted targets were further analyzed using the Gene Ontology and Kyoto Encyclopedia of Genes and Genomes databases. Several functional categories and signaling pathways were enriched, including the \"fc epsilon ri signaling pathway,\" \"endometrial cancer,\" and \"mTOR signaling pathway.\" Expression patterns of 10 differentially expressed miRNAs were verified by reverse transcription-quantitative polymerase chain reaction in mice skins. The results agreed with the results of microarray analysis. These findings suggest that differentially expressed miRNAs revealed by microarray are potential markers for forensic molecular diagnosis of antemortem burns.

Diabetic nephropathy (DN) is a major cause of end-stage renal disease, in which the SMAD signaling pathway plays an important role. The aim of the present study was to identify differentially expressed microRNAs (miRNAs) during the progression of DN and to investigate a selected miRNA in relation to SMAD3/4 and its therapeutic efficacy. The miRNA microarray was used to identify differentially expressed miRNAs in db/db DN mice. Reverse transcription-quantitative polymerase chain reaction and immunoblot analyses were used to detect SMAD3/4 expression. The development of DN in the db/db mice was demonstrated by glucose dysregulation and typical morphological changes in the kidney. miRNA-346 (miR-346) was identified as one of the differentially expressed miRNAs. The expression of SMAD3/4 was significantly attenuated by miR-346 administration and the therapeutic effects of miR-346 were observed in the DN mouse models. miR-346 was identified as a negative regulator of SMAD3/4. SMAD3/4 was upregulated in the renal tissue of db/db mice. The administration of miR-346 attenuated the SMAD3/4 expression in renal tissue and ameliorated the renal function and glomerular histology in DN mice. This study paves the way for clinical studies of miR-346 in DN.

The aim of this study is to examine the microRNA (miRNA) expression of epithelial ovarian cancer (EOC) in both drug-resistant and drug-sensitive tissues and to explore the pathogenic characteristics of drug-resistant miRNAs in EOC. The samples with 10 cases of drug-resistant and drug-sensitive EOC tissue were obtained from undergoing surgical resection of ovarian cancer (OC). Total miRNAs were extracted and isolated, respectively. Hybridization was carried out on miRNA microarray chip. Real-time polymerase chain reaction (RT-PCR) was performed to confirm the difference of miRNA expression. Bioinformatic software was used to predict the possible target genes of each miRNA which expressed differently. The results indicated that four miRNAs related drug-resistance been identified, and the expression of hsa-miR-152 and hsa-miR-381 in drug-resistant OC tissue was significantly higher compared with those in drug-sensitive tissue ( P  < 0.01). However, expression of hsa-miR-200a-3p and hsa-miR-429 were downregulated in drug-resistant tissues ( P  < 0.01). The results obtained by miRNA microarrays of differential expression with hsa-miR-106b-3p, hsa-miR-152, hsa-miR-200a-3p, hsa-miR-381, and hsa-miR-429 were confirmed by real-time PCR. There were 62 significantly different miRNAs, including 42 significant upregulated miRNAs and 20 significant downregulated miRNAs in the drug-resistant tissue. Five databases, including Target Scan, miRanda, miRDB, PicTar5, and RNA22, were used for bioinformatics prediction. In conclusion, miRNA microarray analysis has become a fast and efficient molecular biological technology for the study of biological information. hsa-miR-152, hsa-miR-200a-3p, hsa-miR-381, and hsa-miR-429 may participate in the formation of drug resistance in EOC through the target genes predicted.