Project description:We performed microarray miRNA expression profiling of diabetes induced rat via intraperitoneal (I.P) administration of streptozotocin (STZ). Rats were considered diabetic when their blood glucose exceeded 200 mg/dL (11 mmol/L).
Project description:Urinary exosomal miRNA profiling was conducted in urinary exosomes obtained from 8 healthy controls (C), 8 patients with type II diabetes (T2D) and 8 patients with type II diabetic nephropathy (DN) using Agilent´s miRNA microarrays.
Project description:The aim of this study was to compare miRNA expression in urinary exosomes from type 1 diabetic patients with and without incipient diabetic nephropathy. Overnight urine collections were obtained from normo- and microalbuminuric type 1 diabetic patients. Urines were pre-cleared by both centrifugation and filtration, urinary exosomes were isolated by two consecutive ultracentrifugation steps and total RNA extracted. Differential miRNA profiling was performed using a Human TaqMan miRNA Array A on an 7900HT Fast Real-Time PCR System. Results showed that expression of 22 urinary exosomal miRNAs differed in incipient diabetic nephropathy. Differential qPCR miRNA profiling was performed on urinary exosomes from 2 pairs of micro/normoalbuminuric type 1 diabetic patients comparable for age, sex, diabetes duration, and tightly matched for HbA1c (1° pair: 8.1 vs. 8.1, 2° pair: 8.7 vs. 8.7). MiRNAs were considered differentially expressed if they exhibited greater than twofold expression differences in both pairs.
Project description:Diabetes mellitus is a complex metabolic disorder. Long-term hyperglycemia may induce diabetic keratopathy, which is mainly characterized by decreased tear secretion, damaged innervation, weakened cell junctions, and impaired wound healing responses. To investigate the differential expressed genes in the regulation of diabetic keratopathy, we established streptozocin-induced diabetic and non-diabetic control male Brown Norway rats. Total RNA was extracted from the corneal epithelium rats, and were subjected to whole-transcriptome sequencing analysis. Firstly, Cutadapt was used to remove the reads that contained adaptor contamination, low quality bases and undetermined bases. Then sequence quality was verified using FastQC(http://www.bioinformatics.babraham.ac.uk/projects/fastqc/). We used Bowtie2 and Hisat2to map reads to the genome from corneal epithelium of streptozocin-induced diabetic and non-diabetic control rats. The mapped reads of each sample were assembled using String Tie. Then, all transcriptomes from corneal epithelium Samples were merged to reconstruct a comprehensive transcriptome using perl scripts. We then performed gene expression profiling analysis using data obtained from RNA-seq of 6 corneas from STZ-induced diabetic rats and 6 corneas from normal controls.
Project description:The purpose of this study was the identification of RNAs contained in the urinary exosome (UExo) from dogs and cats. The quality of total RNA in isolated urinary exosome (UExo)-derived total RNAs obtained from the column-based method (urine 1 mL) was checked by using a Bioanalyzer, and samples from normal renal function (NR) group and kidney disease (KD) group were pooled as one sample for each group. We collected NR dogs (n = 37), KD dogs (n = 47), NR cats (n=43), and KD cats (n = 45). For the next generation sequencing, libraries were prepared according to the manufacturer’s protocols and sequenced using 50-base reads acquired by using a HiSeq 2000 platform. The December 2011 (GRCm38/mm10) mouse (Mus musculus) genome data were used as reference. As a result, we could identify the miRNA from these samples.
Project description:The aim of this study was to compare miRNA expression in urinary exosomes from type 1 diabetic patients with and without incipient diabetic nephropathy. Overnight urine collections were obtained from normo- and microalbuminuric type 1 diabetic patients. Urines were pre-cleared by both centrifugation and filtration, urinary exosomes were isolated by two consecutive ultracentrifugation steps and total RNA extracted. Differential miRNA profiling was performed using a Human TaqMan miRNA Array A on an 7900HT Fast Real-Time PCR System. Results showed that expression of 22 urinary exosomal miRNAs differed in incipient diabetic nephropathy.
Project description:This study is to identify urinary exosome microRNAs (miRNAs) that are unique to premature ovarian insufficiency (POI) with and without Turner syndrome and to use them as diagnostic markers for POI patients. We examined the miRNAexpression profile in urine exosomes from POI patients with and without Turner syndrome.
Project description:Diabetes mellitus (DM) is a leading cause of chronic kidney disease and the pathobiology of diabetic nephropathy is widely studied. Less, however, is known about urinary bladder disease in DM despite dysfunctional voiding being a common clinical problem. We hypothesised that diabetic cystopathy would have a characteristic molecular signature, due to the adaptive response to increased urine load combined with the metabolic impacts of DM. To distinguish the consequences of DM from polyuria we compared bladders of untreated control, diabetic (streptozotocin-induced) and sucrose-treated male Wistar rats after 16 weeks using gene array