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Background: Age-related macular degeneration (AMD) is the leading cause of vision loss in the developed world and the detection of its onset and progression are based on retinal morphological assessments. MicroRNA (miRNA) have been explored extensively as biomarkers for a range of neurological diseases including AMD, however differences in experimental design and the complexity of human biology have resulted in little overlap between studies. Using preclinical animal models and clinical samples, this study employs a novel approach to determine a serum signature of AMD progression. Methods: Serum miRNAs were extracted from mice exposed to photo-oxidative damage (PD; 0, 1, 3 and 5 days), and clinical samples from patients diagnosed with reticular pseudodrusen or atrophic AMD. The expression of ~800 miRNAs was measured using OpenArray™, and differential abundance from controls was determined using the HTqPCR R package followed by pathway analysis with DAVID. MiRNA expression changes were compared against quantifiable retinal histological indicators. Finally, the overlap of miRNA changes observed in the mouse model and human patient samples was investigated. Results: Differential miRNA abundance was identified at all PD time-points and in clinical samples. Importantly, these were associated with inflammatory pathways and histological changes in the retina. Further, we were able to align findings in the mouse serum to those of clinical patients. Conclusion: In conclusion, serum miRNAs are a valid tool as diagnostics for the early detection of retinal degeneration, as they reflect key changes in retinal health. The combination of pre-clinical animal models and human patient samples led to the identification of a preliminary serum miRNA signature for AMD. This study is an important platform for the future development of a diagnostic serum miRNA panel for the early detection of retinal degeneration.

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Cysticercus pisiformis (C. pisiformis), the larval form of Taenia pisiformis, parasitize mainly the liver, omentum and mesentery of rabbits and cause huge economic losses in the rabbit breeding industry. MicroRNA (miRNA), a short non-coding RNA, is widely and stably distributed in the plasma and serum. Numerous data demonstrates that, after parasitic infection, miRNAs become the key regulatory factor for controlling host biological processes. However, the roles of serum miRNAs in C. pisiformis-infected rabbits have not been elucidated. In this study, we compared miRNA expression profiles between the C. pisiformis-infected and healthy rabbit serum using RNA-seq. A total of 192 miRNAs were differentially expressed (fold change ≥ 2 and p < 0.05), including 79 up- and 113 downregulated miRNAs. These data were verified by qRT-PCR (real time quantitative polymerase chain reaction) analysis. Additionally, GO analysis showed that the target genes of these dysregulated miRNAs were most enriched in cellular, single-organism and metabolic processes. KEGG pathway analysis showed that these miRNAs target genes were involved in PI3K-Akt, viral carcinogenesis and B cell receptor signaling pathways. Interestingly, after aligning clean reads to the T. pisiformis genome, four (miR-124-3p_3, miR-124-3p_4, miR-124a and novel-miR1) T. pisiformis-derived miRNAs were found. Of these, novel-miR1was upregulated in different periods after C. pisiformis infection, which was verified qRT-PCR, and pre- novel-miR-1 was amplified from the cysticerci by RT-PCR, implying novel-miR-1 was derived from C. pisiformis and has great potential for the diagnosis of Cysticercosis pisiformis infection. This is the first investigation of miRNA expression profile and function in the serum of rabbits infected by C. pisiformis, providing fundamental data for developing diagnostic targets for Cysticercosis pisiformis.

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BACKGROUND: We aimed to develop a diagnostic indicator of stroke based on serum miRNAs correlated to systolic blood pressure. METHODS: Using miRNA expression profiles in GSE117604 from the Gene Expression Omnibus (GEO), we utilized the WGCNA to identify hub miRNAs correlated to systolic blood pressure (SBP). Differential analysis was applied to highlight hub differentially expressed miRNAs (DE-miRNAs), whereby we built a miRNA-based diagnostic indicator for stroke using bootstrap ranking Least Absolute Shrinkage and Selection Operator (LASSO) regression with 10-fold cross-validation. The classification value of the indicator was validated with receiver operating characteristic (ROC) analysis in both the training set and test set, as well as quantitative real-time PCR (qRT-PCR) for the feature miRNAs. Further, target genes of hub miRNAs and hub DE-miRNAs were retrieved for functional enrichment. RESULTS: A total of 447 hub miRNAs in the blue modules were significantly correlated with systolic blood pressure (r = 0.32, false discovery rate = 10-6). Target genes predicted with the hub miRNAs were mostly implicated in the Kyoto Encyclopedia of Genes and Genomes (KEGG) terms including mitogen-activated protein kinase (MAPK) pathway, senescence, and TGF-ß signaling pathway. The diagnostic indicator with miR-4420 and miR-6793-5p showed remarkable performance in the training set (area under curve [AUC]= 0.953), as well as in the test set (AUC = 0.894). Results of qRT-PCR validated the diagnostic value of the two miRNAs embedded in the proposed indicator. CONCLUSIONS: We developed a panel of two miRNAs, which is a good diagnostic indicator for stroke. These results require further investigation.

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Parkinson's disease (PD) is the second most common neurodegenerative disease worldwide, being largely characterized by motor features. MicroRNAs (miRNAs) are small non-coding RNAs, whose deregulation has been associated with neurodegeneration in PD. In this study, miRNAs targeting cell death and/or inflammation pathways were selected and their expression compared in the serum of PD patients and healthy controls. We used two independent cohorts (discovery and validation) of 20 idiopathic PD patients (iPD) and 20 healthy controls each. We also analyzed an additional group of 45 patients with a mutation in the leucine-rich repeat kinase 2 (LRRK2) gene (LRRK2-PD). miRNA expression was determined using Taqman qRT-PCR and their performance to discriminate between groups was assessed by receiver operating characteristic (ROC) curve analysis. We found miR-146a, miR-335-3p, and miR-335-5p downregulated in iPD and LRRK2-PD patients versus controls in both cohorts. In addition, miR-155 was upregulated in LRRK2-PD compared to iPD patients showing an appropriate value of area under the ROC curve (AUC 0.80) to discriminate between the two groups. In conclusion, our study identified a panel of inflammatory related miRNAs differentially expressed between PD patients and healthy controls that highlight key pathophysiological processes and may contribute to improve disease diagnosis.

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OBJECTIVE: To investigate the clinical significance of serum let-7a-5p and miR-21-5p in the diagnosis of breast cancer. METHODS: We examined 32 healthy people and 30 patients with benign breast lesions as controls and 90 breast cancer patients as study subjects. The expression of let-7a-5p and miR-21-5p were detected in all subjects' samples, and Cel-miR-39-3p was used as a spike-in reference. Serum miRNAs were extracted by the TRIzol method, and reverse transcription was performed with specific primers for let-7a-5p, miR-21-5p and Cel-miR-39-3p, and 2-µL reverse transcription products were used as PCR templates. A SLAN-96P fluorescent quantitative PCR instrument was used for quantitative PCR detection. RESULTS: (1) The serum levels of carcinoembryonic antigen (CEA)and carbohydrate antigen 15-3 (CA15-3) in the breast cancer group were higher than those in the healthy controls and patients with benign breast lesions; (2) The expression level of let-7a-5p in the serum of the breast cancer group was lower than that in the healthy control group (P<0.05), but there was no significant difference compared to the breast benign lesion group (P>0.05); (3) The serum expression level of miR-21-5p in the breast cancer group was lower than that in the healthy control group (P<0.05) but was not significantly different from that in the patients with benign breast lesions (P>0.05). CONCLUSION: Reduced expression of Let-7a-5p and miR-21-5p levels is of little value for early diagnosis of breast cancer; however reduced expression of Let-7a-5p and miRNA-21-5p may serve as non-invasive biomarkers for the diagnosis of breast cancer metastasis, and combination of these markers with CEA and CA15-3 can help to distinguish benign breast lesions from breast cancer.

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Objective@#To investigate the expression levels and clinical significance of serum miRNAs, including miR-146a-5p, miR-155-5p and miR-124a-3p, in patients with rheumatoid arthritis (RA). @*Methods@#The clinical data, whole blood and serum samples from 39 RA patients hospitalized in our hospital during October 2016 and October 2018 were collected. Whole blood specimens were used to determine erythrocyte sedimentation rate (ESR), and serum samples were used to detect rheumatoid factor (RF), C-reaction protein (CRP) and miRNAs. The expression levels of serum miR-146a-5p, miR-155-5p and miR-124a-3p in RA patients were detected by SYBR Green real-time fluorescence quantitative PCR. @*Results@#The expression levels of serum miR-146a-5p (1.742±1.058) in RA patients before treatment were significantly higher than that in healthy controls (HC, 1.045±0.772), which were positively correlated with DAS28 scores (r=0.836 5,P=0.004 5), ESR (r=0.437 2, P=0.032 5) and RF levels (r=0.733 6,P=0.013 7). However, the expression levels of serum miR-155-5p (U=42.00,P=0.032 9) and miR-124a-3p (U=44.5,P=0.044 5) in RA patients were significantly lower than that in HC, and the expression levels of serum miR-155-5p were negatively correlated with RF levels (r=-0.445 3,P=0.031 6), and the expression levels of serum miR-124a-3p were negatively correlated with DAS28 scores (r=-0.538 7,P=0.025 8) and RF levels (r=-0.436 5,P=0.046 3). After treatment, the expression levels of serum miR-146a-5p (U= 60.00,P=0.003 8) in RA patients were significantly decreased, while the expression levels of serum miR-155-5p (U=64.00,P=0.005 9) and miR-124a-3p (U=85.00,P=0.042 2) were significantly increased. @*Conclusion@#The abnormal expression levels of serum miR-146a-5p, miR-155-5p and miR-124a-3p in RA patients have potential clinical significance for the diagnosis of RA.

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Circulating microRNAs (miRNAs) are emerging as promising diagnostic biomarkers for autism spectrum disorder (ASD), but their usefulness for detecting ASD remains unclear. Nowadays, development of promising biomarkers for ASD remains a challenge. Recently, dysregulation of the miRNAs expression in postmortem brain tissue, serum and peripheral blood, have been associated with ASD. Circulating miRNAs are known to be secreted by a number of different cells and can interpose delivery of information into receiver cells, thus affecting their functions. Based on this fact, it is supposed that serum miRNAs could be a novel class of biomarkers for prognosis or diagnosis of pathological disorders including ASD. In the current research, we investigated whether the expression patterns of circulating miRNAs showed dysregulation in subjects diagnosed with ASD. Expression levels of serum miR-328-3p and miR-3135a were analyzed by quantitative reverse transcription polymerase chain reaction (qRT-PCR) method of subjects diagnosed with ASD in comparison with healthy control subjects. Our data showed that miR-328-3p and miR-3135a were substantially down-regulated in ASD patients than in those of healthy control subjects. Moreover, target gene analysis of altered serum miRNAs displayed that these molecules targeted 162 genes denoted as unique validated targets in the miRWalk database, 71 of which appear to participate in biological pathways involved in synaptic pathways and neurodegenerative condition such as Alzheimer, Huntington and Parkinson diseases. Finally, the results strongly suggested that dys-regulated serum miRNAs might be involved in molecular pathways associated with ASD and miR-328-3p and miR-3135a have the potential to be promising novel biomarkers for ASD.

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BACKGROUND: Colon cancer, one of the most common causes of cancer-related deaths, arises from adenomatous polyps. In these years, circulating microRNAs (miRNAs) have attracted increasing attention as novel biomarkers for colon cancers. The dysregulated circulating miRNAs in patients with colon adenomas has not been well-understood. METHODS: Here, we aimed to identify miRNA profile in the serum of patients with colon adenomas or colon cancer by using microarray. Then we validated eight differentially expressed miRNAs (DEMs) by qRT-PCR and predicted their targets. RESULTS: We identified 26 DEMs from Adenomas versus Normal comparison (11 up-regulations and 15 down-regulations), 72 DEMs from Cancer versus Normal comparison (19 up-regulations and 53 down-regulations) and 17 DEMs from Cancer versus Adenomas comparison (4 up-regulations and 13 down-regulations). Moreover, three DEMs identified from Cancer versus Normal comparison were included in the list of DEMs identified from Cancer versus Adenomas comparison, and may be specific diagnostic biomarkers for colon cancer. Five down-regulated miRNAs identified from Cancer versus Normal comparison were included in the list of DEMs identified from Adenomas versus Normal comparison, and may be important for the development of colon polyps and cancer. CONCLUSIONS: We discovered 8 circulating miRNAs associated with colon adenomas and colon cancer, and these miRNAs may potentially serve as noninvasive screening biomarkers for colon cancer. Our study is useful for expanding our understanding in the development of colon adenomas and colon cancer, and thus provide novel insights into colon cancer pathogenesis and prevention.

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OBJECTIVES: The aim of this study was to compare the expression levels of serum miRNAs in diabetic retinopathy and type 2 diabetes mellitus. METHODS: Serum miRNA expression profiles from diabetic retinopathy cases (type 2 diabetes mellitus patients with diabetic retinopathy) and type 2 diabetes mellitus controls (type 2 diabetes mellitus patients without diabetic retinopathy) were examined by miRNA-specific microarray analysis. Quantitative real-time polymerase chain reaction was used to validate the significantly differentially expressed serum miRNAs from the microarray analysis of 45 diabetic retinopathy cases and 45 age-, sex-, body mass index- and duration-of-diabetes-matched type 2 diabetes mellitus controls. The relative changes in serum miRNA expression levels were analyzed using the 2-ΔΔCt method. RESULTS: A total of 5 diabetic retinopathy cases and 5 type 2 diabetes mellitus controls were included in the miRNA-specific microarray analysis. The serum levels of miR-3939 and miR-1910-3p differed significantly between the two groups in the screening stage; however, quantitative real-time polymerase chain reaction did not reveal significant differences in miRNA expression for 45 diabetic retinopathy cases and their matched type 2 diabetes mellitus controls. CONCLUSION: Our findings indicate that miR-3939 and miR-1910-3p may not play important roles in the development of diabetic retinopathy; however, studies with a larger sample size are needed to confirm our findings.

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BACKGROUND: Circulating miRNAs have been demonstrated as biomarkers for a number of diseases. In the present study, we aimed to compare the serum levels of the miRNA-221-3p, miRNA-382-5p, and miRNA-4271 between ischemic stroke and healthy control subjects and explore the potential roles as noninvasive biomarkers in the diagnosis of ischemic stroke. METHODS: Seventy-eight patients with ischemic stroke (60 ± 10.47 years old) and 39 healthy control subjects (61 ± 5.14 years old), age and sex matched, were recruited into the present study. The circulating miRNA levels were determined by quantitative real-time polymerase chain reaction using miRNA qPCR Assay Kit (CW Biotech, Beijing, China). Receiver operating characteristic (ROC) curves were analyzed using the SPSS software package (version 17) (SPSS Inc., Chicago, IL). RESULTS: Circulating serum miRNA-221-3p and miRNA-382-5p levels were significantly lower in patients with ischemic stroke compared to healthy control subjects, whereas there was no significant difference in the serum levels of miRNA-4271 between the stroke patients and healthy controls (P > .05). ROC curves revealed the areas under the curve for circulating miRNA-221-3p, miRNA-382-5p, and miRNA-4271 to be .8106, .7483, and .6317 in ischemic stroke patients compared with healthy volunteers, respectively. There were no correlations between circulating miRNAs and laboratory determinations except that the levels of circulating miRNA-4271 were positively correlated with glucose (r = .274, P = .031). CONCLUSIONS: Our findings suggest that serum circulating miRNA-221-3p and miRNA-382-5p might be used as potential noninvasive biomarkers for the diagnosis of ischemic stroke.

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BACKGROUND: Given the challenges of confirming prenatal alcohol exposure (PAE) during pregnancy using currently established biomarkers of alcohol consumption, we examined whether serum microRNAs (miRNAs) may serve as stable biomarkers for PAE. Alterations in the levels of specific circulating miRNAs have been associated with various disease states and in animal models of fetal alcohol spectrum disorder. METHODS: Pregnant women in this prospective study were recruited from substance abuse and general maternity clinics affiliated with the University of New Mexico. Serum was collected at the time of admission for delivery from 14 subjects who reported ≥1 binge-drinking episode or ≥3 drinks/wk during pregnancy and 16 subjects who reported abstinence during pregnancy and tested negative for 5 ethanol biomarkers. Total RNA was isolated from serum and used for microarray analysis. RESULTS: False discovery rate-corrected analyses of covariance revealed that 55 miRNAs were significantly altered between the 2 groups. Hierarchical clustering using only the significantly altered miRNAs grouped samples into alcohol-consuming and non-alcohol-consuming individuals. Discriminant analysis then identified miRs-122*, -126, -216b, -221*, -3119, -3942-5p, -4704-3p, -4743, -514-5p, and -602 as the top 10 discriminators between the 2 groups. Ingenuity Pathway Analysis of putative miRNA targets illustrated that miRNAs identified in this study are involved in biological pathways that mediate the effects of alcohol, such as brain-derived neurotrophic factor, ERK1/2, and PI3K/AKT signaling. CONCLUSIONS: This is the first report of alterations in serum miRNA expression that are associated with alcohol use during human pregnancy. These results suggest that serum miRNAs could be useful as biomarkers of alcohol exposure.

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Multiple myeloma (MM) originates from malignant plasma cells, leading to multiple destructive lytic bone lesions that occur in more than 80% of MM patients. MicroRNAs have been reported to be involved in development of bone lesions in MM. However, the circulating microRNA as diagnostic and prognostic biomarkers for bone lesions has not been elucidated yet. In this study, we identified differentially expressed miRNAs that are potentially involved in myeloma-related bone disease in serum of MM patients. MiR-214 and miR-135b was shown to be increased in serum of MM patients with bone lesions. Serum level of miR-214 and miR-135b was highly correlated with the severity of lytic bone lesions and demonstrated as a diagnostic tool for identifying bone diseases based on results of a receiver operating characteristic analysis (ROC). In addition, patients with high levels of serum miR-214 had a dismal survival with significantly shortened progression free survival (PFS) and overall survival (OS). Interestingly, bisphosphonates treatment significantly extended PFS and OS in patients with higher level of miR-214 comparing to patients without bisphosphonates treatment. Taken together, our findings revealed the significance of circulating miR-214 and miR-135b levels in detection of bone disease and in prediction of prognosis of patients with multiple myeloma, suggesting its potential clinical applications. The result of this study also set the foundation for searching more circulating miRNA as biomarker for tumor bone lesions.

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Gliomas are derived from astroglial precursors or astrocytes, accounting for 40 % central nervous system tumors. MicroRNAs (miRNAs) are a class of endogenous, small (19- to 23-nucleotides) non-coding RNAs involved in cancer progression. Recent studies show that circulating miRNAs are associated with the clinicopathological features and prognosis of gliomas. Serum miRNAs may serve as novel biomarkers for gliomas diagnosis. This review explores the possibilities of using serum miRNAs as prognostic, diagnostic markers, and therapeutic targets in gliomas.

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Multiple myeloma (MM) is the second most common hematologic malignancy characterized by the clonal expansion of plasma cells. Despite continuing advances, novel biomarkers are needed for diagnosis and prognosis of MM. In our study, we characterized the diagnostic and prognostic potential of circulating microRNAs (miRNAs) in MM. Serum miRNA levels were analyzed in 108 newly diagnosed symptomatic MM patients and 56 healthy donors (HDs). Our analysis identified 95 dysregulated miRNAs in newly diagnosed MM patients. Of the 95 dysregulated miRNAs, dysregulation of miR-19a, miR-92a, miR-214-3p, miR-135b-5p, miR-4254, miR-3658 and miR-33b was confirmed by quantitative reverse transcription PCR (RT-qPCR). Receiver operating characteristic analysis revealed that a combination of miR-19a and miR-4254 can distinguish MM from HD with a sensitivity of 91.7% and specificity of 90.5%. Decreased expression of miR-19a was positively correlated with international staging system advancement, del(13q14) and 1q21 amplification. Furthermore, downregulation of miR-19a resulted in significantly decreased progression-free survival (PFS) and overall survival (OS). Our analysis indicated that the poor prognostic correlation of miR-19a expression was independent of genetic abnormalities in MM. Multivariate analysis revealed that miR-19a was a significant predictor of shortened PFS and OS. Interestingly, although miR-19a levels portend a poor prognosis, patients with low miR-19a levels had an improved response to bortezomib compared to those with high miR-19a profile. Patients with downregulated miR-19a experienced a significantly extended survival upon bortezomib-based therapy. These data demonstrate that the expression patterns of serum microRNAs are altered in MM, and miR-19a levels are a valuable prognostic marker to identify high-risk MM.

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The purpose of our study was to screen preliminary differential expression bone-related microRNAs (miRNAs) in serum of patients with osteogenesis imperfacta and to clarify whether serum microRNA is a promising biomarker for osteogenesis imperfecta. geNorm and several other programes were performed to select suitable reference genes for quantitative detection of serum miRNAs from 6 candidate control genes. With geometric averaging of selected reference genes as a normalization factor, fluorescence-based quantitative real-time reverse transcriptase-polymerase chain reaction (qRT-PCR) was used to detect expression levels of more than 100 bone-related miRNAs obtained by means of miRanda, Targetscan and Pictar software calculations and reading the literature. Through analysis of expression stability and pairwise variations, all 6 candidate reference genes had a stable expression level in serum of 8 healthy controls and 8 patients with different characrteristics, and the optimal number of reference genes for normalization was 4 (snRNAU6, miR-92a, miR-16, and Let-7a). For further validation, the expression stability of 4 reference genes remained steady in serum of another 8 healthy controls and 16 patients with osteogenesis imperfecta (M < 1.5). When normalized using multiple control genes, 11 bone-related miRNAs showed differential expression in serum of 8 osteogenesis imperfecta patients compared with 8 healthy controls. In conclusion, we identified snRNAU6, miR-92a, miR-16, and Let-7a as an internal reference gene group for qRT-PCR normalization and screening results revealed that there existed many differential expression bone-related miRNAs in serum of patients with osteogenesis imperfecta compared with healthy controls, and that these miRNAs had potential to be biomarkers for serologic tests and diagnosis of osteogenesis imperfecta with analysis of bioinformation.