Table 1.
Primer sequences.
Fig 1.
Experimental technical route.
Table 2.
Clinical data characteristics of the experimental group and control group.
Fig 2.
Expression levels of hsa_circ_0004099 in healthy subjects and patients with AIS.
A: The Mann-Whitney U-test was used for two-sample comparisons. The expression level of hsa_circ_0004099 was lower in patients with AIS compared with that in healthy subjects. B: The Steel-Dwass test was used for pairwise comparisons. The levels of hsa_circ_0004099 in patients with small-artery occlusion (SAO), large-artery arteriosclerosis (LAA), and cardiac-origin cardioembolism (CE) were different compared to those of normal individuals (P<0.001). C: Comparison of National Institutes of Health Stroke Scale (NIHSS) disease severity scores and hsa_circ_0004099 levels using Spearman rank correlation: 1 on the abscissa represents "mild,” 2 represents "moderate,” and 3 represents "severe." The expression level of hsa_circ_0004099 showed a downward trend and was highly negatively correlated with the NIHSS scores (P<0.001, r = -0.7053). D: Pearson correlation analysis was used to compare the infarct time with the hsa_circ_0004099 levels: 1 grid on the abscissa represents 1 hour, and each circle represents a patient with AIS (40 patients in total). As shown in the figure, with the prolongation of infarction time, the expression of hsa_circ_0004099 showed a downward trend. The levels of hsa_circ_0004099 were negatively correlated with the infarct time (P<0.001, r = -5130).
Fig 3.
Receiver operating characteristic (ROC) curve.
The abscissa indicates the sensitivity, while the ordinate indicates 1-specificity. At a cut-off of 0.85, the area under the curve (AUC) peaked, while the sensitivity and specificity of the best cut-off point were 0.95 and 0.90, respectively. At this time, the hsa_circ_0004099 level was the highest predictive value for AIS risk.
Fig 4.
Downstream of hsa_circ_0004099 intersecting microRNAs (miRNAs) and mRNAs.
A: Ten intersecting miRNAs predicted using the circular RNA Interactom and circBank. B: Ten miRNAs predicted 616 intersecting mRNAs. C: CSCD identified the miRNA-response elements (MRE), RNA-binding proteins (RBP), and open reading frames (ORF) of hsa_circ_0004099, which obtained the binding score of hsa_circ_0004099 and the intersection of 10 miRNAs; the closer the value was to 100, the higher was the confidence.
Fig 5.
Circular RNA-microRNA-mRNA (circRNA-miRNA-mRNA) network.
Inverted triangles represent hsa_circ_0004099, triangles represent miRNAs, while circles represent mRNAs.
Fig 6.
A: GO enrichment analysis: BP, biological process; CC, cellular component; MF, molecular function. The abscissa represents the number of enriched genes, while the ordinate describes the enriched items. The darker the bar color, the more significant the difference between the enriched items. B: KEGG enrichment analysis: the abscissa represents the number of enriched genes, while the ordinate represents the enriched pathway entries. Many cancer pathways and some common pathways are shown. The size of the circle represents the number of enriched genes: the larger the circle, the higher the number of enriched genes, and the redder the color, the closer the relationship with the enriched entry.
Fig 7.
Kyoto encyclopedia of genes and genomes (KEGG) signaling pathway map.
Red indicates genes that are enriched in cancer signaling pathways. The four genes, TCF7L2, NRAS, CTNNB1, and KRAS, framed in yellow are jointly enriched by these two pathways and are mainly involved in cell proliferation.
Fig 8.
Protein-protein interaction (PPI) network.
SMAD4, HIF1A, CTNNB1, CDKN1B, CDK6, FOXO3, KRAS and NRAS are the group of core proteins screened using NetworkAnalyzer.