Introduction
Methodology
Ethical considerations
Study settings and subjects
Study tools
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Detailed medical history from caregivers, with particular emphasis on the developmental history, history of allergic or autoimmune manifestations, and the family history of allergic or autoimmune diseases.
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Clinical examination, including general, chest, cardiac, abdominal, and neurological assessments, to ensure the inclusion and exclusion criteria.
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Neuropsychiatric assessment, the degree of the disease severity was assessed using the Childhood Autism Rating Scale (CARS) [16], which rates the child on a scale from one to four in each of fifteen behavioral items. Scores up to 30 indicate mild autism, above 30 to 37 indicate moderate autism, and from 37 to 60 indicate severe autism.
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Assessment of IQ level, using full-scale IQ (FSIQ) [17], derived from the sum of all tasks in the Stanford-Binet 5. The test encompasses both the verbal and nonverbal domains of cognitive ability and offers an overall overview of the current state of general intellectual functioning.
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Assessment of serum levels of IL-17 A and IL-22, three mL of whole blood samples were withdrawn from all participants under complete aseptic conditions into a plain vacutainer tube containing gel and clot activator for serum separation by centrifugation at 3000 RPM for 10 min after whole blood clotting for 20 min. Separated sera were kept at -80°C until the time of analysis of IL-17 A and IL22 by human enzyme-linked immune sorbent assay (ELISA) kits (Bioassay Technology LLC, china, cat.No. E0142Hu and E0038Hu) The results were plotted on standard curves. The kit detection range of IL-17 A was from 2 pg/mL to 600 pg/mL, and the minimum detectable level of human IL-17 A was 1.06 pg/mL. Meanwhile, the kit detection range of IL-22 was from 5 pg/mL to 1500 pg/mL, and the minimum detectable level of human IL-22 was 2.48 pg/mL.
Statistical analysis
Results
ASD Patients | ||
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n = 24 | ||
Type of Autism | Regressive | 0 (0.0%) |
Primary | 24 (100.0%) | |
Childhood Autism Rating Scale (CARS) | Mean ± SD | 33.46 ± 4.83 |
Range | 25–44 | |
Severity groups according to CARS scale | Mild up to 30 | 6 (25.0%) |
Moderate 30–37 | 13 (54.2%) | |
Severe 37–60 | 5 (20.8%) | |
Score of intelligence quotient (IQ) test | Mean ± SD | 61.58 ± 9.30 |
Range | 40–80 | |
Below average intelligence | 5(20.8%) | |
Cognitive impairment | 19 (79.2%) | |
Electroencephalogram (EEG) abnormalities | Negative | 21 (87.5%) |
Positive | 3 (12.5%) | |
Presence of convulsions | No | 21 (87.5%) |
Yes | 3 (12.5%) | |
Frequency of convulsions | Irregular | 2 (66.7%) |
Every 3 months | 1 (33.3%) | |
Controlled convulsions | Yes | 3 (100.0%) |
IL-17 pg/mL | Test value | p-value | IL-22 pg/mL | Test value | p-value | ||
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Median (IQR) | Median (IQR) | ||||||
Gender | Female | 255 (160–380) | -0.536• | 0.592 | 1110 (800–1150) | -1.708• | 0.088 |
Male | 265 (250–320) | 700 (660–900) | |||||
Presence of convulsions | No | 260 (250–320) | -0.132• | 0.895 | 850 (700–1000) | -1.535• | 0.125 |
Yes | 320 (210–320) | 700 (400–700) | |||||
Electroencephalogram (EEG) abnormalities | Negative | 260 (250–320) | -0.132• | 0.895 | 850 (700–1000) | -1.535• | 0.125 |
Positive | 320 (210–320) | 700 (400–700) |
Childhood Autism Rating Scale (CARS) severity | Test value | p-value | ||||
---|---|---|---|---|---|---|
Mild | Moderate | Severe | ||||
n = 6 | n = 13 | n = 5 | ||||
Age (years) | Median (IQR) | 5.25 (5–10) | 5 (5–8) | 7 (6–8) | 1.062≠ | 0.588 |
Range | 413 | 3.59 | 3.514 | |||
Gender (n., %) | Female | 3 (50.0%) | 1 (7.7%) | 2 (40.0%) | 4.677* | 0.096 |
Male | 3 (50.0%) | 12 (92.3%) | 3 (60.0%) | |||
Presence of convulsions (n., %) | No | 6 (100.0%) | 11 (84.6%) | 4 (80.0%) | 1.213* | 0.545 |
Yes | 0 (0.0%) | 2 (15.4%) | 1 (20.0%) | |||
Frequency of convulsions (n., %) | Irregular | 0 (0.0%) | 2 (100.0%) | 0 (0.0%) | 3.000* | 0.083 |
Every 3 months | 0 (0.0%) | 0 (0.0%) | 1 (100.0%) | |||
Controlled convulsions | Yes | 0 (0.0%) | 2 (100.0%) | 1 (100.0%) | – | – |
Family history of allergy (n., %) | No | 6 (100.0%) | 12 (92.3%) | 5 (100.0%) | 0.883* | 0.643 |
Yes (Asthma) | 0 (0.0%) | 1 (7.7%) | 0 (0.0%) | |||
Family history of autoimmune diseases (n., %) | No | 6 (100.0%) | 11 (84.6%) | 5 (100.0%) | 1.846* | 0.397 |
Yes | 0 (0.0%) | 2 (15.4%) | 0 (0.0%) | |||
Score of intelligence quotient (IQ) test | Mean ± SD | 65.83 ± 8.82 | 62.69 ± 8.47 | 53.60 ± 8.59 | 3.011• | 0.071 |
Range | 55–80 | 40–70 | 41–65 | |||
Electroencephalogram (EEG) abnormalities (n., %) | Negative | 6 (100.0%) | 11 (84.6%) | 4 (80.0%) | 1.213* | 0.545 |
Positive | 0 (0.0%) | 2 (15.4%) | 1 (20.0%) | |||
Interleukin (IL)-17 (pg/mL) | Median (IQR) | 262.5 (250–320) | 260 (220–300) | 360 (320–380) | 2.349≠ | 0.309 |
Range | 120–520 | 170–440 | 160–440 | |||
Interleukin (IL)-22 (pg/mL) | Median (IQR) | 750 (660–850) | 700 (600–900) | 1120 (920–1150) | 4.448≠ | 0.108 |
Range | 240–1450 | 400–1100 | 700–1400 |
IL-17 | IL-22 | |||
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r | p-value | r | p-value | |
Interleukin (IL)-17 A (pg/mL) | – | – | 0.524** | 0.009 |
Interleukin (IL)-22 (pg/mL) | 0.524** | 0.009 | – | – |
Age (years) | 0.219 | 0.305 | 0.067 | 0.757 |
Childhood Autism Rating Scale (CARS) scale | 0.097 | 0.654 | 0.432** | 0.035 |
Scores of intelligence quotient (IQ) test | 0.116 | 0.589 | -0.142 | 0.507 |