Demographic data of the subject
The study included 50 samples from the case group and 50 samples from the control group. As Table 1 shows, there were no significant differences in diabetes, smoking, age, sex, alcohol consumption, weight, and height between the two groups (p > 0.05). However, there were notable differences in hypertension and Body mass index (BMI) between the two groups (p <0.05). And 54% of patients with IS (27 patients) had large vessel ischemic stroke, while 46% of patients with IS (23 patients) had small vessel ischemic stroke.
Table 1
Demographic characteristics of the study population.
Variable
|
Total
|
Control
|
Case
|
P-value*
|
Gender N (%)
Female
Male
|
45
55
|
23 (51.1%)
27 (49.1%)
|
22 (49%)
28 (51 %)
|
0.50
|
Age, Mean (SD)
|
|
54.16 (14.2)
|
58.46 (10.5)
|
0.89
|
History of Cigarette, N (%)
Yes
No
|
26
74
|
11 (42.3%)
39 (52.7%)
|
15 (57.7%)
35 (47.3%)
|
0.247
|
History of Alcohol, N (%)
Yes
No
|
17
83
|
6 (35.3%)
44 (53.0%)
|
11 (64.7%)
39 (47.0%)
|
0.143
|
History of Diabetic disease, N (%)
Yes
No
|
29
71
|
12 (41.4%)
38 (53.5%)
|
17 (58.6%)
33 (46.5%)
|
0.189
|
History of Hypertension, N (%)
Yes
No
|
42
58
|
15 (35.7%)
35 (60.3%)
|
27 (64.3%)
23 (39.7%)
|
0.013
|
Height, Mean (SD)
|
|
167.42 (9.5)
|
167.68 (9.7)
|
0.89
|
Weight, Mean (SD)
|
|
69.70 (12.4)
|
70.2 (12.7)
|
0.84
|
BMI, Mean (SD)
|
|
20.67 (1.6)
|
24.63 (3.6)
|
<0.001
|
*T-test and chi-square test were used. |
Evaluation of the serum levels of biochemical factors
As Table 2 shows, there are significant differences in serum levels of ATG5, Apo B-48, TG, cholesterol, and oxidative stress markers such as MDA, TOS and TAC between both groups. However, no remarkable differences were found in serum levels of LDL-C and HDL-C between both groups.
Table 2
Comparison of the blood levels of the biochemical parameters between case and control groups
Variable
|
Control
Mean (SD)
|
Patients with a stroke
Mean (SD)
|
P-value*
|
Apo B48(μg/mL)
|
4.09 (0.35)
|
4.67 (0.26)
|
<0.0001
|
ATG5(ng/mL)
|
18.60 (3.2)
|
29.72 (6.2)
|
<0.0001
|
MDA (nmoL/mL)
|
1.60 (0.30)
|
1.97 (0.46)
|
<0.0001
|
TOS(μmoL/L)
|
10.72 (1.14)
|
11.89 (1.62)
|
<0.0001
|
TAC (mmoL/L)
|
1.23 (0.22)
|
1.48 (0.29)
|
<0.0001
|
LDL-C(mg/dL)
|
105.92 (37.5)
|
112.90 (49.0)
|
0.426
|
HDL-C(mg/dL)
|
46.26 (9.4)
|
44.62 (10.8)
|
0.422
|
CHOL (mg/dL)
|
172.28 (38.9)
|
192.54 (58.4)
|
0.044
|
TG (mg/dL)
|
152.64 (57.1)
|
200.38 (64.4)
|
<0.0001
|
T-test was used.
TG: Triglyceride, LDL-C: Low-density lipoprotein cholesterol, CHOL: Cholesterol, HDL-C: High–density lipoprotein cholesterol
|
*
Evaluation of ATG5, Apo B-48, MDA, TOS, and TAC Serum Levels
As Figure 1 shows, serum levels of ATG5 were significantly increased (29.72 ± 6.2 ng/ml) compared to the control group (18.60 ± 3.2 ng/ml) with p 0.0001. Serum levels of Apo B-48 were significantly increased in the case group (4.67 ± 0.26 μg/ml) compared to the control group (4.09 ± 0.35 μg/ml) with p 0.0001. Serum levels of MDA were significantly increased in the case group (1.97 ± 0.46 nmol/ml) compared to the control group (1.60 ± 0.30 nmol/ml) with p< 0.0001. Serum levels of TOS were significantly increased in the case group (11.89 ± 1.62 μmol/l) compared to the control group (10.72 ± 1.14 μmol/l) with p< 0.0001. Serum TAC levels were significantly elevated in the case group (1.48 ± 0.29 mmol/l) compared to the control group (1.23 ± 0.22 mmol/l) with p< 0.0001.
logistic regression analysis
As shown in Table 3, logistic regression analysis revealed that with a one-unit increase in serum levels of Apo B-48, ATG5, MDA, TOS, and TAC, the odd of developing IS increased 905-, 2-, 15-, 1.8-, and 53-fold, respectively, which is statistically significant (P< 0.05).
Table 3
The relationship between the blood levels of Apo B48, ATG5, and stress biomarkers and Stroke disease by Logistic regression
Variables
|
B
|
S. E
|
Odds ratio
|
95 % CI
|
P-value*
|
Apo B48(μg/mL)
|
6.808
|
1.328
|
905.375
|
67.016 to 12231.446
|
<0.0001
|
ATG5(ng/mL)
|
0.712
|
0.156
|
2.039
|
1.501 to 2.769
|
<0.0001
|
MDA (nmoL/mL)
|
2.734
|
0.692
|
15.388
|
3.960 to 59.790
|
<0.0001
|
TOS(μmoL/L)
|
0.600
|
0.167
|
1.822
|
1.314 to 2.527
|
<0.0001
|
TAC (mmoL/L)
|
3.985
|
1.002
|
53.792
|
7.547 to 383.402
|
<0.0001
|
*Logistic regression was used. |
Analyzing ROC curve
To investigate the diagnostic value of the above serum factors, a ROC curve analysis is required. As shown in Table 4, the area under the ROC curve (AUC) values for ATG5, Apo B-48, MDA, TOS, and TAC were 0.9656, 0.9120, 0.7530, 0.7102, and 0.7588 of 95%, respectively. The results were statistically significant at p 0.05 because the area under the curve of the results (95% confidence interval) did not have an AUC value of 0.5. The largest AUC belongs to ATG5 and Apo B-48, indicating these two parameters are more suitable for diagnosing IS (Figure 2). Table 5 shows the cutoff values for ATG5, Apo B-48, MDA, TOS, and TAC based on the Youden index results. According to the Youden index, the best cutoff value for Apo B-48 is a serum level of ≥4.35, indicating that serum levels above 4.35 indicate patients with IS. Serum levels below 4.35 indicate healthy individuals. Additionally, the sensitivity is 84%, and the specificity is 80% at this cutoff value, so 84% of patients with IS and 80% of healthy individuals is correctly diagnosed as patients with IS and healthy individuals, respectively. Also, the cutoff value of ATG5 in serum ≥23.5 shows a sensitivity of 90% and a specificity of 96% in diagnosing patients with IS and healthy individuals, respectively (Table 5).
Table 4
The AUC analysis for Apo B48, ATG5, and stress biomarkers in IS patients.
P-Value
|
95% CI
|
Std. Error
|
Area
|
Variable(s)
|
<0.0001
|
0.8589 to 0.9651
|
0.02708
|
0.9120
|
Apo B48(μg/ml)
|
<0.0001
|
0.9341 to 0.9971
|
0.01607
|
0.9656
|
ATG5(ng/mL)
|
<0.0001
|
0.6585 to 0.8475
|
0.04821
|
0.7530
|
MDA (nmoL/mL)
|
0.0003
|
0.6086 to 0.8118
|
0.05181
|
0.7102
|
TOS(μmoL/L)
|
<0.0001
|
0.6644 to 0.8532
|
0.04814
|
0.7588
|
TAC (mmoL/L)
|
Table 5
The sensitivity and specificity of the Apo B48, ATG5, and stress biomarkers in the detection of stroke according to the Youden index
Variable
|
Cutoff point value
|
sensitivity
|
specificity
|
Apo B48(μg/mL)
|
≥4.35
|
0.84
|
0.80
|
ATG5(ng/mL)
|
≥23.5
|
0.90
|
0. 96
|
MDA (nmoL/mL)
|
≥1.85
|
0.62
|
0.76
|
TOS(μmoL/L)
|
≥10.32
|
0.84
|
0.42
|
TAC (mmoL/L)
|
≥ 1.235
|
0.80
|
0.48
|
Spearman's correlation
The results of the Spearman correlation test showed there was a small but statistically nonsignificant association between serum levels of ATG5, Apo B-48, MDA, TOS, and TAC in patients with IS (p > 0.05) (Table 6).
Table 6
The correlation between serum concentration of the Apo B48, ATG5, and stress biomarkers in the detection of stroke
Variable
|
N
|
Correlation coefficient
|
P-value*
|
Apo B48(μg/mL)
|
50
|
-0.070
|
0.629
|
ATG5(ng/mL)
|
50
|
0.081
|
0.576
|
MDA (nmol/mL)
|
50
|
-0.203
|
0.158
|
TOS(μmol/L)
|
50
|
0.045
|
0.757
|
TAC (mmol/L)
|
50
|
0.032
|
0.828
|
Logistic regression analysis
An analysis of the correlation between serum levels of Apo B-48, ATG5, MDA, TOS, and TAC with vascular involvement in patients with IS by logistic regression analysis revealed there was no statistically significant correlation between the serum levels of these factors and vascular involvement in IS patients (P >0.05) (Table 7).
Table 7
The relationship between the blood levels of Apo B48, ATG5, and stress biomarkers and type of vessel involvement¥ in patients with stroke disease.
Variable
|
B
|
S. E
|
Odds ratio
|
95 % CI
|
P-value*
|
Apo B48(μg/mL)
|
1.395
|
1.116
|
4.033
|
0.452 to 35.958
|
0.212
|
ATG5(ng/mL)
|
0.092
|
0.052
|
1.096
|
0.991 to 1.213
|
0.075
|
MDA (nmoL/mL)
|
-1.295
|
0.739
|
0.274
|
0.064 to 1.166
|
0.080
|
TOS(μmoL/L)
|
-0.138
|
0.178
|
0.871
|
0.614 to 1.235
|
0.438
|
TAC(mmoL/L)
|
-0.119
|
0.959
|
0.888
|
0.135 to 5.820
|
0.901
|
* Logistic regression was used. ¥ type of vessel involvement: Large vessel and small vessel. |