Study On The Difference Of Two Non-coding RNAs Expression In Coronary Heart Disease And Moderate Cognitive Impairment Plasma

PartⅠ Differential Expression of Circular RNA in Plasma of Patients with Coronary Heart DiseaseObjective:Circular RNA（circRNA）is a type of closed circular RNA.It belongs to non-coding RNAs（ncRNAs）classification.The circular RNA have no 5’end cap structure and 3’terminal poly（A）tail srtuctrue,and is not easily affected by RNA exonuclease.So compared with linear RNA structure,circRNA is more stable.In recent years,related studies have shown that circular RNA is not only widely present in eukaryotes,but also has a variety of biological functions and plays an important regulatory role.Studies have shown that circular RNA is closely related to the occurrence and development of various diseases.circRNA can be detected extensively in human body tissues and even in body fluids such as blood and saliva.The expression of circRNA in various organs and tissues in human body is different.It is with the characteristics of tissue-specific.The above-mentioned characteristics of circular RNA can be used as a diagnostic marker of diseases,and related literatures have reported that circular RNA can serves as the biomarker for diseases such as tumors.However,the research of circRNA that uesed as biomarker for cardiovascular disease（CVD）is relatively insufficient.In the previous study of our team,bioinformatics analysis techniques were used to screen out a number of circRNAs that were differentially expressed in human vascular smooth muscle cells.In this study,we selected several circRNAs with significantly expression from them and studied their expression in peripheral blood of patients with coronary atherosclerosis by real-time fluorescence quantitative PCR（qRT-PCR）.And we further explored whether there are differences expression in the plasma between patients with coronary atherosclerosis and healthy controls.This study aims to find potential diagnostic biomarkers of coronary atherosclerotic disease,and provides novel directions and ideas for the clinical diagnosis and assessment of coronary atherosclerotic disease.Methods:We collected a total of 23 blood samples from patients diagnosed with coronary atherosclerosis and blood samples from 24 healthy controls at the same time.Real-time fluorescence quantitative analysis was used to detect the expression levels of hsa＿circ＿0007888,hsa＿circ＿0000219,hsa＿circ＿0000721in the blood of patients with coronary atherosclerosis.hsa＿circ＿0007888,hsa＿circ＿0000219,hsa＿circ＿0000721 were calculated in the plasma of patients with coronary atherosclerosis and healthy people by the 2^-△△Ctmethod to express difference of multiples.Data processing was performed using two independent sample t-tests for statistical analysis.Results:1.hsa＿circ＿0007888,hsa＿circ＿0000219,hsa＿circ＿0000721 are expressed in the peripheral blood of patients with coronary atherosclerosis.2.The expression of hsa＿circ＿0007888,hsa＿circ＿0000219,hsa＿circ＿0000721 in the peripheral blood of patients with coronary atherosclerosis are significantly higher than that in health controls.and the expression up-regulation multiples are 7.605、7.532、3.555 times respectively.Both differential expression are statistically significant（P<0.05）.Conclusions:hsa＿circ＿0007888,hsa＿circ＿0000219,hsa＿circ＿0000721 are up-regulated significantly in peripheral blood of patients with coronary atherosclerosis,and hsa＿circ＿0007888,hsa＿circ＿0000219,hsa＿circ＿0000721can serve as potential clinical diagnostic biomarkers for coronary atherosclerotic disease.PartⅡ Study on MicroRNA expression in the plasma of patients withmoderate cognitive impairmentObjective:MicroRNAs（miRNAs）are endogenous non-coding RNAs（ncRNAs）,which are commonly found in human body fluid.In recent years,a large number of miRNAs have been detected in living organisms.Although miRNAs do not directly participate in the coding of proteins in vivo,they can control the expression of related genes by inhibiting target genes or degrading target genes,so they have very important biological significance.In recent years,related studies have shown that miRNA can not only participate in cell differentiation,metabolism,aging and other life activities,but also has close relationship with the development of human diseases such as cancer,diabetes,and nervous system diseases.Alzheimer disease（AD）is a neurodegenerative disease and there is currently no effective cure.Related studies have shown that miRNA may be involved in the development of Alzheimer’s disease.A large number of abnormally expressed miRNAs were found in the hippocampus,cerebrospinal fluid,and blood of Alzheimer’s patients.Therefore,miRNA may be used as a potential diagnostic biomarker for Alzheimer’s disease,and there have also been reported in some researchers.However,is there abnormal expression of miRNA in the plasma of MCI patients with a diabetic background?Whether miRNA can be served as an early biomarker for AD diagnosis?There is no relevant research be reported.Therefore,in this study,we aimed to find miRNAs which are abnormally expressed in the plasma of MCI patients with diabetes backgroud and search for potential diagnostic markers.In this experiment,the expression levels of seven kinds of miRNAs were detected in the plasma of MCI patients with diabetes backgroud by real-time fluorescence quantitative（qRT-PCR）.Our further study explored whether these 7 miRNAs have different expression levels in the plasma between MCI patients with diabetes backgroud and patients in the control group.This study may provide a new idea and direction for the early diagnosis and evaluation of Alzheimer’s disease.Methods:Seven miRNAs are detected in this study:hsa-miR-9,hsa-miR-29a,hsa-miR-29b,hsa-miR-106,hsa-miR-137,hsa-miR-146a,andhsa-miR-181c.We collected a total of 5 blood samples from MCI patients with diabetes background（test group）and 8 blood samples from patients with diabetes（control group）in hospital neurology department.Real-time fluorescence quantitative technology is used to detect the expression levels of the above 7miRNAs in patients.The expression difference mutiples of 7 miRNAs:hsa-miR-9,hsa-miR-29a,hsa-miR-29b,hsa-miR-106,hsa-miR-137,hsa-miR-146a,and hsa-miR-181c are calculated using the relative quantitation2^-ΔΔCt method.Experimental data is processed with two independent samples t test for statistical analysis.Results:1.Seven miRNAs:hsa-miR-9,hsa-miR-29a,hsa-miR-29b,hsa-miR-106,hsa-miR-137,hsa-miR-146a,and hsa-miR-181c can be expressed in the plasma of MCI patients who have a diabetic background.2.There are no statistically significant differences in the expression of 7miRNAs:hsa-miR-9,hsa-miR-29a,hsa-miR-29b,hsa-miR-106,hsa-miR-137,hsa-miR-146a,and hsa-miR-181c in the blood samples between patients in the case group and in the control group（P>0.05）.Conclusions:There is no significant expression difference in hsa-miR-9,hsa-miR-29a,hsa-miR-29b,hsa-miR-106,hsa-miR-137,hsa-miR-146a,hsa-miR-181c in the plasma between moderate cognitive impairment with diabetes backgroud and diabetes.