Objective To investigate the changes of circulating miRNAs expression profiles in different subtypes of ischemic stroke according to the Trial of Org 10 172 in Acute Stroke Treatment.Methods We selected 16 patients diagnosed as acute ischemic stroke at first time in the Department of Neurology,the Affiliated Hospital of Medical College Qingdao University from November to December in 2012.They were divided into large artery atherosclerosis (LAA) stroke group (n =8) and small artery occlusive (SAO) stroke group (n =8).At the same time,8 healthy checkup subjects were selected as control.High-throughput sequencing was used to detect the expression profiles of miRNAs in the plasma,and the high-throughput sequencing results were validated by quantitative real-time polymerase chain reaction.We performed the miRNAs variance analysis and associated bioinformatics analysis.Results Thirty hundred and sixty-nine miRNAs were detected in LAA group,SAO group and the control group.We found remarkable differences (fold change ＞2,P ＜0.01) in the expression of 12 miRNAs,including let-7a-5p,miR-744-5p,etc.,between any two groups of the three groups.Thirty-four miRNAs,containing miR-126-5p,miR-23a-3p,miR-143-3p,etc.,had a lower expression (fold change ＞2,P ＜0.01)in LAA group than that in the control group.In comparison with the control group,miR-1304-3p and miR-451a were significantly down-regulated (fold change ＞ 2,P ＜ O.01),while 27 miRNAs were significantly up-regulated (fold change ＞2,P ＜0.01) in SAO group.The expression levels of miR-146b-5p,miR-23a-3p and miR-451 a were validated in accordance with the results of real-time PCR.Target gene prediction and functional analysis revealed that target genes regulated by differential expression of miRNAs were mainly associated with cell proliferation,adhesion,metabolism and other biological functions.Conclusion miRNAs are differently expressed in the plasma of LAA group,SAO group and healthy control group,which suggest that miRNAs might play different roles in the pathogenesis of LAA stroke and SAO stroke by regulating downstream target genes.

Objective To investigate the difference of expression profiling of plasma miRNAs (microRNA) between the patients with small artery occlusive stroke (SAO) and the healthy subjects.Methods Eight patients with SAO classified by TOAST were selected and 8 healthy subjects were used as a control group.High-throughput sequencing technology was used to detect the expression profiling of plasma miRNAs.The differentially expressed miRNAs were screened.Real-time fluorescent quantitative polymerase chain reaction was used to validate the results,and the target gene prediction and bioinformatics analysis were performed.Results The miRNA difference analysis showed that the expression profilings of miRNA-127,miRNA-99b-5p,miRNA-320,and other 19 miRNAs in the SAO group were significantly upregulated compared with those in the control group (all P＜0.01),while miRNA-451a and other 5 miRNAs in the SAO group were significantly downregulated compared with those in the control group (all P ＜0.01).The validated results of miRNA-127,miRNA-99b-5p,miRNA-320,and miRNA-451a with real-time fluorescent quantitative polymerase chain reaction were consistent with those of the high-throughput sequencing.Bioinformatics analysis showed that the miRNA-regulated target genes expressed differentially were mainly correlated with cell proliferation,adhesion,phylogenetic development,macromolecule metabolism,and other biological functions.Conclusions There are significant differences in the expression profiling of plasma miRNAs between the patients with SAO and the healthy subjects,suggesting that miRNAs may play a regulatory role via target genes in pathogenesis of SAO.

Objective To investigate the relationships of plasma alpha2-macroglobulin (α2-MG) level with occurrence of small-artery occlusion (SAO) and cognitive function in SAO patients.Methods Subjects were collected in our hospital from December 2012 to September 2013;58 patients with SAO stroke,56 with large artery atherosclerosis (LAA) classified by Trial of Org 10172 in Acute Stroke Treatment (TOAST),as well as 58 healthy controls,were enrolled.The serum α2-MG levels were measured by enzyme-linked immunosorbent assay (ELISA) and compared among the three groups.The receiver operating characteristic (ROC) curve was drawn to evaluate the value of α2-MG as a biomarker for SAO.White matter lesions (WMLs) were assessed by MRI using Fazekas classification,and mini-mental state examination (MMSE) was used to evaluate the cognitive function.The correlations of plasma α2-MG level with both Fazekas scores and MMSE scores were analyzed.Results The serum α2-MG levels in SAO patients ([6.70±3.17] μg/mL) were significantly higher than those in LAA patients ([5.04±1.93] μg/mL) and controls ([4.86±1.68] μg/mL) (P＜0.05).Binary Logistic regression analysis showed that the plasma α2-MG level was an independent risk factor for occurrence of SAO (OR=1.409,P=0.001;OR=1.443,P=0.004).The ROC curves obtained that α2-MG=4.23 μg/mL was the cutoff values for SAO,enjoying sensitivity up to 86.2％.SAO patients with different severities of WMLs had significantly different serum α2-MG levels (F=26.509,P=0.000);the serum α2-MG levels were positively correlated with Fazekas scores (r=0.733,P=0.000) and negatively correlated with MMSE scores (r=-0.679,P=0.000).Conclusions Serum α2-MG level is an independent risk factor for SAO and might be a biomarker for SAO.Furthermore,serum α2-MG level,being associated with high-grade WMLs,might contribute to evaluating the cognitive impairment in SAO patients.