CARDIOVASCULAR JOURNAL OF AFRICA • Volume 34, No 2, May/June 2023 100 AFRICA to check the normality for continuous variables, followed by the test for homogeneity of variances, while the chi-squared test was used to categorise the variables. The independent sample t-test was used for continuous variables consistent with normal distribution and homogeneity of variance. If variables were not consistent with normal distribution and homogeneity of variance, the Kruskal–Wallis H-test was used; p < 0.05 was considered statistically significant. All statistical analyses were performed using SPSS 20.0 (SPSS Inc, Chicago, IL, USA). Results The participants in this study were divided into two cohorts. The clinical and demographic characteristics are summarised in Tables 1 and 2. Micro-array analysis of the expression profiles of circRNAs was performed using the Agilent human circRNA array (v2.0). The expression levels of circRNAs varied significantly between the two groups (Fig. 2). A total of 2 884 circRNAs was detected with differential expression. Of these, 1 989 were upregulated and 895 were downregulated in the type II cardio-renal syndrome group (Table 3). Differentially expressed mRNAs between patients with type II cardio-renal syndrome and healthy controls were enriched in two pathways, including haematopoietic cell lineage and cell adhesion molecules. To obtain the important circRNA that would play a major role in type II cardio-renal syndrome, the candidates that were to be verified genes were selected from the 1 989 upregulated circRNAs utilising stricter screening criteria: fold change > 6 and p < 0.0001, and from the 895 downregulated circRNAs utilising stricter screening criteria: fold change > 2.6 and p < 0.0003. We selected hsa_cir_0001763. Table 2. Comparision of the clinical and demographic characteristics of the participants in this study Characteristics Control group (n = 12) CRS group (n = 15) p-value Male gender 7 (58) 9 (60) 1.00 Age (years) 36.00 ± 8.12 79.00 ± 5.55 0.000 Hypertension 3 10 0.05 Coronary heart disease 5 9 0.45 Smoker 7 9 1.00 Height (cm) 160.58 ± 4.79 161.53 ± 7.15 0.697 Weight (kg) 59.70 ± 6.25 65.00 ± 13.27 0.186 BMI (kg/m2) 23.13 ± 1.97 24.89 ± 4.82 0.215 SBP (mmHg) 107.66 ± 9.56 140.00 (135.00–140.00) 0.000 DBP (mmHg) 67.58 ± 6.51 71.80 ± 14.77 0.333 ALT (U/l) 12.00 (11.25–16.75) 14.00 (9.00–19.00) 0.897 AST (U/l) 17.05 (15.77–20.70) 21.50 (16.60–26.00) 0.142 TC (mmol/l) 4.70 ± 0.86 3.99 ± 0.90 0.059 TG (mmol/l) 1.06 (0.82–1.47) 1.42 ± 0.50 0.157 HDL-C (mmol/l) 1.25 ± 0.29 1.11 ± 0.33 0.284 LDL-C(mmol/l) 2.56 ± 0.64 2.29 ± 0.64 0.310 NT-proBNP (pg/ml) 90.95 (73.67–107.75) 2037 (1153–4156.5) 0.000 eGFR (ml/min/1.73 m2) 107.40 (101.13–112.56) 54.43 (42.46–58.30) 0.000 CRS, type II cardio-renal syndrome; BMI, body mass index; SBP, systolic blood pressure; DBP, diastolic blood pressure; ALT, alanine aminotransferase; AST, aspartate aminotransferase; TC, total cholesterol; TG, triglyceride; HDL-C, high-density lipoprotein cholesterol; LDL-C, low-density lipoprotein cholesterol; NT-proBNP, N-terminal pro-B-type natriuretic peptide. Table 1. Comparision of the clinical and demographic characteristics of the participants in this study Characteristics Control group (n = 5) CRS group (n = 20) p-value Male gender, n (%) 3 (60) 12 (60) 1.00 Age (years) 40.80 ± 7.19 76.90 ± 7.77 0.000 Hypertension 0 9 0.12 Coronary heart disease 0 10 0.06 Smoker 3 11 1.00 Height (cm) 161.20 ± 4.43 164.45 ± 9.28 0.459 Weight (kg) 54.40 ± 3.13 66.65 ± 13.32 0.001 BMI (kg/m2) 20.95 ± 1.34 24.56 ± 4.18 0.074 SBP (mmHg) 106.00 ± 7.96 161.20 ± 4.43 0.001 DBP (mmHg) 69.40 ± 6.02 74.35 ± 12.15 0.391 ALT (U/l) 12.00 (10.50–29.00) 12.50 (10.00–20.25) 0.413 AST (U/l) 23.28 ± 6.93 19.20 (17.20–27.00) 0.708 TC (mmol/l) 4.46 ± 0.95 4.25 ± 0.68 0.591 TG (mmol/l) 0.95 (0.79–2.49) 1.31 (1.03–1.68) 0.126 HDL-C (mmol/l) 1.31 ± 0.43 1.15 ± 0.21 0.250 LDL-C (mmol/l) 2.36 ± 0.72 2.48 ± 0.49 0.661 NT-proBNP (pg/ml) 80.15 (55.00–121.5) 1462 (945–3122.5) 0.000 eGFR (ml/min/1.73 m2) 106.54 (94.38–109.45) 48.69 (41.05–56.76) 0.000 CRS, type II cardio-renal syndrome; BMI, body mass index; SBP, systolic blood pressure; DBP, diastolic blood pressure; ALT, alanine aminotransferase; AST, aspartate aminotransferase; TC, total cholesterol; TG, triglycerides; HDL-C, high-density lipoprotein cholesterol; LDL-C, low-density lipoprotein cholesterol; NT-proBNP, N-terminal pro-B-type natriuretic peptide. D5 E4 E3 E5 E2 E1 D1 C8 B4 A4 D3 D2 A5 B1 A1 B2 C5 D4 B5 B3 C1 C3 A2 C4 A3 Colour Key –3 –2 –1 0 1 2 3 Value Fig. 2. Heat map of circRNA micro-array profile in control individuals (n = 5) and cardio-renal syndrome patients (n = 20). The expression of circRNAs is hierarchically clustered on the y-axis; the corresponding circRNAs are shown on the right. Blood samples from control individuals and cardio-renal syndrome patients are hierarchically clustered on the x-axis. Expression values are presented in red and green to indicate upregulation and downregulation, respectively. Numbers with E indicate control individuals, and numbers with A–D indicate CAD patients.
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