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BACKGROUND: It has been recently discovered that vitamin D receptor is widely distributed in the female reproductive system such as the ovary and uterus, which is closely related to adverse pregnancy outcomes such as spontaneous abortion and gestational diabetes mellitus, but its specific mechanism is still unclear. OBJECTIVE: To investigate potential differentially expressed miRNAs and related regulatory networks via a whole transcriptome analysis of the uterine tissue of vitamin D-deficient mice. METHODS: Eighteen female C57BL/6J mice were randomly divided into two groups: a vitamin D deficiency group and a normal control group. Mice in the vitamin D deficiency group were fed with vitamin D deficiency diet, and mice in the normal control group were fed with vitamin D sufficiency diet. The body mass of the mice was recorded every week. After 8 weeks, the serum levels of 25,(OH)D3 and hormones in the mice were tested. Hematoxylin-eosin staining was used to detect mouse uterine tissue. Uterine tissue samples of two groups were collected for transcriptome sequencing, and bioinformatics analyses of gene ontology and Kyoto encyclopedia of genes and genomes. RESULTS AND CONCLUSION: There was no difference in body mass between the two groups. Compared with the normal control group, the levels of 25,(OH)D3 and estradiol in mouse serum were significantly reduced, and the testosterone level was significantly increased in the vitamin D deficiency group. Hematoxylin-eosin staining of uterine tissue showed a reduction in the endometrial folds of mice in the vitamin D deficiency group. Twenty-five differentially expressed miRNAs were screened out by transcriptome sequencing, of which 9 were up-regulated (such as miR-541-5p and miR-205-5p) and 16 were down-regulated (such as miR-378d and miR-708-5p). At the same time, vitamin D deficient mice showed significantly differences in multiple gene ontology enrichment categories, such as developmental process, cell composition, tissue or biogenesis. In addition, the functional enrichment results of Kyoto genes and genomes encyclopedia showed that differentially expressed genes were mostly related to biochemical metabolic pathways and signal transduction pathways. Differentially expressed genes in these enrichment pathways mainly affected PI3K-Akt signaling pathway, MAPK signaling pathway, insulin signaling pathway, gonadotropin releasing hormone signaling pathway, and oocyte meiosis pathway, all of which were closely related to the regulation of the reproductive system. To conclude, vitamin D deficiency can lead to poor uterine implantation conditions in female mice, which is related to multiple signaling pathways, such as insulin regulation, in mouse reproductive system. [ABSTRACT FROM AUTHOR]

Background: Vitiligo is an acquired skin depigmentation disease. It can be misdiagnosed at an early stage and tend to relapse. Serum markers are essential to monitoring the progression of vitiligo. Exosomal miRNAs act as the communication mediator between melanocytes and immune cells. Our study aimed to use serum exosomal miRNAs as a reference for evaluating vitiligo progression. Methods: The miRNAs were extracted from the serum exosomes of ten progressive vitiligo patients (before and after treatment) and ten healthy individuals. We profiled miRNAs expression by RNA sequencing and screened out potential miRNAs and plotted their receiver operating characteristic (ROC) curves to explore their sensitivity and specificity as prognostic biomarkers in vitiligo progression. We examined the correlation between miRNA expression and the lesion area. Different databases were used to predict gene targets of miRNAs, which were analyzed by gene ontology and Kyoto encyclopedia of genes and genomes (KEGG). Results: Our results showed that 141 miRNAs were differentially expressed in serum exosomes of progressive vitiligo patients, and 365 miRNAs were differentially expressed in these patients after treatment compared to healthy individuals. The expression of hsa-miR-487b-3p was significantly lower in these patients compared to healthy individuals. Still, there was no difference in its levels in patients after corticosteroid treatment compared to healthy controls. ROC curve analysis (area under curve = 0.840) indicated that hsa-miR-487b-3p could serve as a biomarker for the prognosis of vitiligo progression. Its expression positively correlated with the lesion area. A total of 41 target genes of hsa-miR-487b-3p were predicted via different databases. KEGG pathways were enriched in phenylalanine metabolism, glycan degradation, and protein export. Conclusion: Serum exosomal hsa-miR-487b-3p can be a biomarker to detect vitiligo progression. The predicted target genes of hsa-miR-487b-3p were enriched in catabolism. Thus, its in progressive vitiligo may accelerate catabolism in melanocytes and cause its impairment. [ABSTRACT FROM AUTHOR]

Seed development is closely related to plant production and reproduction, and MicroRNAs (miRNA) is widely involved in plant development including seed development. Chinese kale, as a Brassicaceae vegetable, mainly depends on seed for proper reproduction. In the present study, Chinese kale seed and silique at different stages were selected to establish small RNA (sRNA) libraries including silique wall sRNA libraries at torpedo-embryo stage (PC), silique wall sRNA libraries at cotyledonary-embryo stage (PD), seed sRNA libraries at torpedo-embryo stage (SC), and seed sRNA libraries at cotyledonary-embryo stage (SD). The results showed that miRNA expressed differentially in the seeds and corresponding siliques at different stages. To further clarify the functional mode of miRNA in the process of seed development, Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis was performed on target genes of the differentially expressed miRNAs, and these target genes were mainly enriched in plant hormone signal transduction, primary and secondary metabolic pathways. After joint analysis with the transcriptome change of the corresponding period, miR156- SPL10 / SPL11 , miR395- APS3 , and miR397- LAC2 / LAC11 modules were identified to be directly involved in the development of Chinese kale seeds. What's more, modified 5′RLM-RACE and Agrobacteria -mediated Chinese kale transient transformation suggest miR395b_2 is involved in sulfur metabolism during seed development by regulating its target gene APS3. [ABSTRACT FROM AUTHOR]

Background: Preeclampsia (PE) is an idiopathic hypertensive disorder of pregnancy and is the leading cause of maternal death, fetal malformation, and premature birth. The purpose of this study is to identify the key molecules and lncRNA-related competitive endogenous (ceRNA) regulatory network in PE. Methods: The differentially expressed mRNAs (DEGs), lncRNAs (DELs), and miRNAs (DEMs) were identified between PE and control using the Deseq R package. In addition, we performed Geno ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) on DEGs and DELs-target genes to explore their function. The ceRNA network was established based on lncRNA-miRNA and miRNA-mRNA interactions and visualized using Cytoscape software. LINCO2532, SLCO4A1-AS1, miR23a-5p, and DYNLRB1 were selected for qRT-PCR assay. Results: Using microarray analysis, we screened 726 DELs (456 upregulated and 370 downregulated), 49 DEMs (37 upregulated and 12 downregulated), and 318 DEGs (230 upregulated and 88 downregulated) between PE patients and control. Based on lncRNA-miRNA pairs and miRNA-mRNA pairs, the ceRNA network was constructed, which contained 16 lncRNA, 1 miRNA (miR-23a-5p), and 1 mRNA (DYNLRB1). LncRNA (LINCO2532 and SLCO4A1-AS1) and DYNLRB1 were downregulated and the expression of miR23a-5p was upregulated in PE patients compared with healthy controls. Conclusion: In this study, the novel ceRNA network was established in the placentas of PE patients. It elucidated the regulatory mechanism of PE, and identified novel PE biomarkers, which have important guiding significance for clinical treatment and further scientific research of PE. [ABSTRACT FROM AUTHOR]

Sichuan mountainous black‐bone (SMB) chicken is a small‐sized black‐feathered chicken breed with low amount of meat, while Dahen (DH) chicken has a larger body size and a faster growth rate. MicroRNAs (miRNAs) are involved in various physiological processes, but their role in chicken muscle growth remains unclear. We aimed to investigate the miRNAs and pathways participating in the muscle growth of chicken. MiRNA profiles of four SMB chickens and four DH chickens were detected by small RNA sequencing. A total of 994 known miRNAs were identified, among which gga‐miR‐1a‐3p, gga‐miR‐148‐3p and gga‐miR‐133a‐3p exhibited the highest enrichment in both breeds of chickens. Thirty‐two miRNAs were differently expressed between SMB and DH chickens. The differently expressed miRNAs were mainly associated with fatty acid metabolism, immunity and MAPK activation‐related processes. Kyoto encyclopaedia of genes and genomes (KEGG) analysis showed that miRNAs were involved in the immunity‐related and MAPK signalling pathways. Moreover, miR‐204 was downregulated in DH chicken compared with SMB chicken, and significantly inhibited the expression of MAP3K13, which is involved in the MAPK pathway. It was confirmed through luciferase reporter assays that miR‐204 specifically inhibited the activity of MAP3K13. Our results helped demonstrate the potential molecular mechanisms of muscle growth in chickens and provide valuable information for chicken breeding. [ABSTRACT FROM AUTHOR]

Background: Oral squamous cell carcinoma (OSCC) is a solid tumor, which originates from squamous epithelium, with about 400,000 new-cases/year worldwidely. Presently, chemoradiotherapy is the most important adjuvant treatment for OSCC, mostly in advanced tumors. However, clinical resistance to chemotherapy still leads to poor prognosis of OSCC patients. Via high-throughput analysis of gene expression database of OSCC, we investigated the molecular mechanisms underlying cisplatin resistance in OSCC, analyzing the differentially expressed genes (DEGs) and their regulatory relationship, to clarify the molecular basis of OSCC chemotherapy resistance and provide a theoretical foundation for the treatment of patients with OSCC and individualized therapeutic targets accurately. Methods: Datasets related to "OSCC" and "cisplatin resistance" (GSE111585 and GSE115119) were downloaded from the GEO database and analyzed by GEO2R. Venn diagram was used to obtain drug-resistance-related DEGs. Functional enrichment analysis and Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis were performed on DEGs using The Database for Annotation, Visualization and Integrated Discovery (DAVID) software. Protein–protein interaction (PPI) network was constructed by STRING (search tool for recurring instances of neighbouring genes) database. Potential target genes of miRNA were predicted via miRDB, and cBioportal was used to analyze the function and survival of the potential functional genes. Results: Forty-eight upregulated DEGs and 49 downregulated DEGs were obtained from the datasets, with cutoff as p < 0.01 and |log FC| > 1. The DEGs in OSCC mainly enriched in cell proliferation regulation, and chemokine activity. In PPI network with hub score > 300, the hub genes were identified as NOTCH1 , JUN , CTNNB1 , CEBPA , and ETS1. Among miRNA–mRNA targeting regulatory network, hsa-mir-200c-3p, hsa-mir-200b-3p, hsa-mir-429, and hsa-mir-139-5p were found to simultaneously regulate multiple hub genes. Survival analysis showed that patients with high CTNNB 1 or low CEBPA expression had poor outcome. Conclusions: In the OSCC cisplatin-resistant cell lines, NOTCH1 , JUN , CTNNB1 , CEBPA , and ETS1 were found as the hub genes involved in regulating the cisplatin resistance of OSCC. Members of the miR-200 family may reverse drug resistance of OSCC cells by regulating the hub genes, which can act as potential targets for the treatment of OSCC patients with cisplatin resistance. [ABSTRACT FROM AUTHOR]

Background: Sepsis is a life-threatening organ dysfunction, initiated by a dysregulated host response to infection. This study aimed to determine key genes and microRNAs (miRNAs) correlated with sepsis. Methods: Three patients with sepsis and three healthy individuals treated as controls were recruited in the current study. To identify differentially expressed mRNAs (DEmRNAs) and differentially expressed miRNAs (DEmiRNAs) between patients with sepsis and controls, RNA-sequencing and bioinformatics analysis were conducted. DEmiRNA-target DEmRNAs analysis and functional annotation of DEmiRNA-target DEmRNAs were performed. Dataset GSE46955, used to validate the expression of selected DEmRNAs, was downloaded from the Gene Expression Omnibus database. Results: Compared with septic patients, a total of 1199 DEmRNAs and 23 DEmiRNAs were identified. Based on DEmiRNA-target DEmRNAs analysis, hsa-miR-106b-5p (degree = 155), hsa-miR-128-3p (degree = 128), and hsa-miR-144-3p (degree = 79) were the top 3 DEmiRNAs that covered most DEmRNAs. The T cell receptor signaling pathway, pathways in cancer, FoxO signaling pathway, and influenza A were the significantly enriched Kyoto Encyclopedia of Genes and Genomes pathways of DEmiRNAtarget DEmRNAs in sepsis. Conclusion: We identified key genes and miRNAs related to sepsis. Our findings will provide new insights into understanding sepsis pathogenesis. [ABSTRACT FROM AUTHOR]

To construct a long noncoding RNA (lncRNA)–microRNA (miRNA)–messenger RNA (mRNA) regulatory network related to epithelial ovarian cancer (EOC) cisplatin‐resistant, differentially expressed genes (DEGs), differentially expressed lncRNAs (DELs), and differentially expressed miRNAs (DEMs) between MDAH and TOV‐112D cells lines were identified. Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were conducted to analyze the biological functions of DEGs. Downstream mRNAs or upstream lncRNAs for miRNAs were analyzed at miRTarBase 7.0 or DIANA‐LncBase V2, respectively. A total of 485 significant DEGs, 85 DELs, and 5 DEMs were identified. Protein–protein interaction (PPI) network of DEGs contrains 81 nodes and 141 edges was constructed, and 25 hub genes related to EOC cisplatin‐resistant were identified. Subsequently, a lncRNA–miRNA–mRNA regulatory network contains 4 lncRNAs, 4 miRNAs, and 35 mRNAs was established. Taken together, our study provided evidence concerning the alteration genes involved in EOC cisplatin‐resistant, which will help to unravel the mechanisms underlying drug resistant. [ABSTRACT FROM AUTHOR]

Background: Although an immense effort has been made to develop a novel biomarker for response to trastuzumab, no reliable biomarkers are available to guide management, expect for HER2. The aim of this study was to examine the relationship between microRNA (miRNA) expression and resistance to trastuzumab. Methods: Differentially expressed miRNAs between trastuzumab‐resistant and trastuzumab‐sensitive cell lines were analyzed using microarrays. We performed Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses to determine the functions of differentially expressed miRNA and their targeted genes. Furthermore, the protein–protein interactions (PPI) network was analyzed. Serum samples were collected from patients with HER2‐positive breast cancer who were treated with trastuzumab. We validated the miRNAs expression levels by quantitative reverse‐transcription polymerase chain reaction (qRT‐PCR) in these serums. Receiver operating characteristic (ROC) curve analysis was performed to evaluate the predictive performance of the miRNA. Results: Using miRNA microarrays, 151 miRNAs that significant differentially expressed between the trastuzumab‐resistant and sensitive cells were identified, including 46 upregulated and 105 downregulated miRNAs. Results of real‐time PCR confirmed seven miRNAs in cell lines. PI3K‐Akt signaling pathway was involved in regulating biological function according to KEGG analysis. Compared with the serums of trastuzumab‐sensitive patients, three miRNAs, namely miR‐200b, miR‐135b, and miR‐29a, were identified to be upregulated, and miR‐224 was downregulated in the trastuzumab‐resistant serums. ROC analysis showed that four miRNAs were correlated with trastuzumab resistance. Furthermore, three subnetwork modules of PPI network were obtained. Conclusion: The results indicated that miRNAs were reliable predictive biomarkers for response to trastuzumab. [ABSTRACT FROM AUTHOR]

Introduction: A great deal of research has reported dysregulated expression of genes in systemic lupus erythematosus (SLE). This study aimed to analyze the lncRNA, miRNA and mRNA expression profile in SLE.Material and Methods: RNA sequencing (RNA-seq) was used to detect the dysregulated RNAs in SLE. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways analysis were used to explore the function of these differentially expressed RNAs.Results: 2,353 lncRNAs, 827 mRNAs and 24 miRNAs were shown to be differentially expressed. GO analyses demonstrated that differentially expressed RNAs were enriched in a variety of molecular functions and biological processes including ribonucleotide, protein serine/threonine kinase activity function, regulation of B cell differentiation and others. KEGG pathway analyses revealed that differentially expressed mRNAs and lncRNAs were both enriched in FcγR-mediated phagocytosis, glycosaminoglycan biosynthesis-chondroitin sulfate/dermatan sulfate and glyoxylate and dicarboxylate metabolism pathways. The up-regulated miRNAs target genes were mainly enriched in the nuclear factor-κB (NF-κB) signaling pathway. The down-regulated miRNAs target genes were significantly enriched in metabolism of xenobiotics by cytochrome P450, bile secretion and terpenoid backbone biosynthesis pathways.Conclusions: The current study reveals a comprehensive expression profile of lncRNAs, miRNAs and mRNAs and implies potential regulatory functions of these RNAs which are involved in the pathogenesis of SLE. [ABSTRACT FROM AUTHOR]

The aim of this study was to explore the expression difference of miRNAs and mRNAs between the follicular phase (FP) and luteal phase (LP) in porcine ovaries and provide a theoretical basis for the research on mammalian reproductive regulation. RNA‐Seq and miRNA‐Seq were used to identify differentially expressed genes (DEGs) and miRNAs (DEMs) between the FP and LP in ovaries of six sows (3‐year‐old Yorkshire pigs with similar weights and same parities). Bioinformatic analysis was used to screen potential genes and miRNAs related to porcine ovarian function. Real‐time qualitative PCR was used to validate the sequencing results. RNA‐Seq results showed that 3,078 genes were up‐regulated, and 1,444 genes were down‐regulated in the LP compared with the FP, and DEGs were significantly enriched in 242 Gene Ontology (GO) terms and 33 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. miRNA‐Seq identified 112 DEMs, of which 25 were up‐regulated and 87 were down‐regulated in the LP compared with the FP. We obtained 186 intersection genes (IGs) between the 4,522 DEGs and 2,444 target genes predicted from the 112 DEMs. After constructing a miRNA‐gene‐pathway network, we identified key miRNAs and genes including miR‐17‐3p, miR‐214, miR‐221‐5p, miR‐125b, FGF1, YWHAG, YWHAZ, FDFT1 and DHCR24, which are enriched in Hippo and PI3K‐Akt signalling pathways, and various metabolic pathways. These results indicate that these key genes and miRNAs may play important roles in the developmental transition from FP to LP in porcine ovaries and represent candidate targets for further study. [ABSTRACT FROM AUTHOR]

The molecular mechanisms underlying neuropathic pain (NP) remain poorly understood. Emerging evidence has suggested the role of microRNAs (miRNAs) in the initiation and development of NP, but the specific effects of miRNAs in NP are largely unknown. Here, we use network‐ and pathway‐based methods to investigate NP‐induced miRNA changes and their biological functions by conducting a systematic search through multiple electronic databases. Thirty‐seven articles meet the inclusion criteria. Venn analysis and target gene forecasting are performed and the results indicate that 167 overlapping target genes are co‐regulated by five down‐regulated miRNAs (rno‐miR‐183, rno‐miR‐96, rno‐miR‐30b, rno‐miR‐150 and rno‐miR‐206). Protein‐protein interaction network analysis shows that 77 genes exhibit interactions, with cyclic adenosine monophosphate (cAMP)‐dependent protein kinase catalytic subunit beta (degree = 11) and cAMP‐response element binding protein 1 (degree = 10) having the highest connectivity degree. Gene ontology analysis shows that these target genes are enriched in neuron part, neuron projection, somatodendritic compartment and nervous system development. Moreover, analysis of Kyoto Encyclopedia of Genes and Genomes reveals that three pathways, namely, axon guidance, circadian entrainment and insulin secretion, are significantly enriched. In addition, rno‐miR‐183, rno‐miR‐96, rno‐miR‐30b, rno‐miR‐150 and rno‐miR‐206 are consistently down‐regulated in the NP models, thus constituting the potential biomarkers of this disease. Characterizing these miRNAs and their target genes paves way for their future use in clinical practice. [ABSTRACT FROM AUTHOR]

[Objectives] This study aims to analyze host miRNAs in the regulation of influenza virus-host interaction using gene chip sequencing technology. [Methods] BALB/c mice were infected with low pathogenic A/Anhui/1/2013 (H7N9) virus and PBS. The lung tissues were collected at day 3 post-infection and were subjected to miRNA chip sequencing to screen differentially expressed miRNA and then verified by RT-qPCR. Targetscan, PITA and microRNAorg programs were used to predict target genes, respectively, and Gene Ontology (GO) and Kyoto Encyclopedia of Gene and Genomes (KEGG) were then used to analyze the biological functions of differential miRNA and their possible signaling pathways. [Resuts] Compared with the PBS infection group, 265 differentially expressed miRNA were detected in the low pathogenic H7N9 infection mice, of which 143 miRNA were significantly up-regulated and 122 miRNA were significantly down-regulated. The differentially expressed miRNA were verified by RT-qPCR, and the results of 10 candidates miRNA were consistent with the chip sequencing results. Further KEGG analysis indicated that these differentially expressed miRNA were mainly enriched in the Rap1 signaling pathway, PI3K-Akt signaling pathway, Hippo signaling pathway, MAPK signaling pathway, Wnt signaling pathway, focal adhesion, autophagy and other immune-related signaling pathways. Combined with differential mRNA information, miRNA-mRNA regulatory network revealed that there were mainly 15 differentially expressed miRNAs and 31 differential target genes enriched into these signaling pathways. [Conclusions] This experiment successfully screened differentially expressed miRNAs caused by low pathogenic H7N9 infection in mice lung tissue, and RT-qPCR proved that it had a good consistency with the expression trend of gene chip sequencing results. This study laid the foundation for further research of the important role of host miRNA in regulating influenza virus replication in mice model. [ABSTRACT FROM AUTHOR]

Background: Patients with dilated cardiomyopathy, increased ventricular volume, pressure overload or dysynergistic ventricular contraction and relaxation are susceptible to develop serious ventricular arrhythmias (VA). These phenomena are primarily based on a theory of mechanoelectric feedback, which reflects mechanical changes that produce alterations in electrical activity. However, very few systematic studies have provided evidence of the preventive effects of artemisinin (ART) on VA in response to left ventricle (LV) afterload increases. MicroRNAs (miRNAs) are endogenous small non-coding RNAs that regulate expression of multiple genes by suppressing mRNAs post-transcriptionally. Aims: The aims of this study were to investigate preventive effects of ART on mechanical VA and the underling molecular mechanisms of differentially expressed miRNAs (DEMs). Methods: For the study, 70 male Wistar rats were randomly divided into seven groups: group 1 was a control group (sham surgery); group 2 was a model group that underwent transverse aortic constriction (TAC) surgery; groups 3, 4, 5 and 6 were administered ART 75, 150, 300 and 600 mg/kg before TAC surgery, respectively; and group 7 was administered verapamil (VER) 1 mg/kg before TAC surgery. A ventricular arrhythmia score (VAS) was calculated to evaluate preventive effects of ART and VER on mechanical VA. The high throughput sequencing-based approach provided DEMs that were altered by ART pretreatment between group 2 and group 4. All predicted mRNAs of DEMs were enriched by gene ontology (GO) and Kyoto Encyclopedia annotation of Genes and Genomes (KEGG) databases. These DEMs were validated by a real time quantitative polymerase chain reaction (RT-qPCR). Results: The average VASs of groups 3, 4, 5, 6 and 7 were significantly reduced compared with those of group 2 (2.70 ± 0.48, 1.70 ± 0.95, 2.80 ± 0.79, 2.60 ± 0.97, 1.40 ± 0.52, vs 3.70 ± 0.67, p < 0.01, respectively). The three top GO terms were neuron projection, organ morphogenesis and protein domain specific binding. KEGG enrichment of the 16 DEMs revealed that MAPK, Wnt and Hippo signaling pathways were likely to play a substantial role in the preventive effects of ART on mechanical VA in response to LV afterload increases. All candidate DEMs with the exception of rno-miR-370-3p, rno-miR-6319, rno-miR-21-3p and rno-miR-204-5p showed high expression levels validated by RT-qPCR. Conclusions: Artemisinin could prevent mechanical VA in response to LV afterload increases. Validated DEMs could be biomarkers and therapeutic targets of ART regarding its prevention of VA induced by pressure overload. The KEGG pathway and GO annotation analyses of the target mRNAs could indicate the potential functions of candidate DEMs. These results will help to elucidate the functional and regulatory roles of candidate DEMs associated with antiarrhythmic effects of ART. [ABSTRACT FROM AUTHOR]

A high incidence of thromboembolic events associated with high mortality has been reported in severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) infections with respiratory failure. The present study characterized post-transcriptional gene regulation by global microRNA (miRNA) expression in relation to activated coagulation and inflammation in 21 critically ill SARS-CoV-2 patients. The cohort consisted of patients with moderate respiratory failure (n = 11) and severe respiratory failure (n = 10) at an acute stage (day 0–3) and in the later course of the disease (>7 days). All patients needed supplemental oxygen and severe patients were defined by the requirement of positive pressure ventilation (intubation). Levels of D-dimers, activated partial thromboplastin time (aPTT), C-reactive protein (CRP), and interleukin (IL)-6 were significantly higher in patients with severe compared with moderate respiratory failure. Concurrently, next generation sequencing (NGS) analysis demonstrated increased dysregulation of miRNA expression with progression of disease severity connected to extreme downregulation of miR-320a, miR-320b and miR-320c. Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis revealed involvement in the Hippo signaling pathway, the transforming growth factor (TGF)-β signaling pathway and in the regulation of adherens junctions. The expression of all miR-320 family members was significantly correlated with CRP, IL-6, and D-dimer levels. In conclusion, our analysis underlines the importance of thromboembolic processes in patients with respiratory failure and emphasizes miRNA-320s as potential biomarkers for severe progressive SARS-CoV-2 infection. [ABSTRACT FROM AUTHOR]

Toxoplasma gondii is an intracellular parasite that infects humans and other warm-blooded animals. Exosomes are endocytic-derived vesicles released by cells, representing an important mode of intercellular communication. In exosomes, specific molecules of proteins, lipids, and mRNAs or miRNAs have been detected, some of which are capable of transferring biologically active molecules to recipient cells. Dendritic cells (DCs) are the only antigen-presenting cells (APCs) that activate the initial immune response. In this study, high-throughput sequencing was used to analyze the exosomal miRNA profile of DC2.4 cells infected with Toxoplasma gondii for 28 h, compared with those of uninfected DC2.4 cells. Differential exosomal miRNAs (DEmiRs) from these two cell groups were analyzed. Through high-throughput sequencing, 3434 DEmiRs were obtained, and 12 stably enriched DEmiRNAs were verified by Reverse Transcription-quantitative Polymerase Chain Reaction (RT-qPCR) and selected for further analysis. The target genes of these 12 miRNAs were predicted with online analysis software and subjected to bioinformatics analyses including protein–protein interaction (PPI) network analysis, key driver analysis (KDA), gene ontology (GO) enrichment, and Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis. These DEmiRs were found to be associated with a variety of biological processes and signaling pathways involved in host ubiquitin system, innate immunity, biosynthesis, and transferase activity and could be potential biomarkers for T. gondii infection. [ABSTRACT FROM AUTHOR]

Background: To explore long-non-coding RNA (lncRNA), microRNA (miRNA) and messenger RNA (mRNA) expression profiles and their biological functions in the urine samples in calcium oxalate (CaOx) patients. Methods: Five CaOx kidney stone patients were recruited in CaOx stone group and six healthy people were included as control group, whose midstream morning urine was collected before the patients were given any medicine on admission. After total RNA was extracted from urine, microarray of miRNA, mRNA and lncRNA were applied to explore their expression variation. Gene ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed to reveal the gene functions of the dysregulated lncRNA-associated competing endogenous RNA (ceRNA) network. Quantitative real-time PCR were performed on HK-2 cells treated with sodium oxalate (NaOx) to further screen out the differentially expression profiles of these RNAs. Results: A total of nine miRNAs, 883 mRNAs and 1002 lncRNAs were differentially expressed in urine of CaOx patients compared with normal population. GO analysis revealed that most of mRNAs from ceRNA network were enriched in terms of respiratory burst, regulation of mitophagy, and protein kinase regulator activity. KEGG pathway analysis of these genes related to ceRNA network highlight their critical role in pentose phosphate pathway, glyoxylate and dicarboxylate metabolism, and Janus kinase/signal transducer and activator of transcription (JAK-STAT) signaling pathway. Five miRNAs (miR-6796-3p, miR-30d-5p, miR-3192–3p, miR-518b and miR-6776-3p), four mRNAs (NT5E, CDH4, CLEC14A, CCNL1) and six lncRNAs (lnc-TIGD1L2–3, lnc-KIN-1, lnc-FAM72B-4, lnc-EVI5L-1, lnc-SERPINI1–2, lnc-MB-6) from the HK-2 cells induced by NaOx were consistent with the expression changes of microarray results. Conclusion: The differential expressed miRNAs, mRNAs and lncRNAs may be associated with numerous variations of the signaling pathways or regulation of metabolism and kinase activity, providing potential biomarkers for early diagnosis of urolithiasis and new basis for further research of urolithiasis mechanism. [ABSTRACT FROM AUTHOR]