| Objective:To analysis the change characteristics of steroid hormones in saliva of prepubertal boys with ASD,and to explore whether steroid hormones can be used as biomarkers for ASD.Methods:1.G-power was used to analyze the minimum sample size.2.From January 2021 to May 2022,Prepubertal boys with ASD under 8years of age who were diagnosed in the Children’s Health Department of Jingyang District Maternal and Child Health Hospital of Deyang City and the Children’s Health Department of Guangyuan Central Hospital according to the DSM-V standard and Autism Diagnostic Observation Schedule-2 scale(ADOS-2)were included in the experimental group.The boys who underwent physical examination in the Children’s Health Care Department of Guangyuan Central Hospital and normal prepubertal boys under 8 years old in Guangyuan Government kindergartens were included as the healthy control group.Indicators related to Child Behavior Scale(CBCL),Autism Behavior Scale(ABC),Social Response Scale(SRS)and Repetitive Behavior Scale-Revised(RBS-R)were collected to assess autistic symptoms.Age,height,and weight were collected to analyze the effects of age,height,and weight on ASD.QTRAP(?) 6500+ LC-MS/MS system was used to analyze the concentrations of the steroid hormones cortisol,dehydroepiandrosterone(DHEA),and pregnenolone in saliva samples from the two groups.Mann Whitney U test,Spearman correlation analysis,logistic regression model,and receiver operating characteristic(ROC)analysis were used to compare the concentrations of cortisol,dehydroepiandrosterone,and pregnenolone among the behaviors of perpubertal boys with ASD.Results:1.G-power results showed that at least 21 cases should be included in each group.We actually collected 55 cases of prepubertal boys with clinically confirmed ASD and 24 cases of normal boys as controls.Therefore,our sample size is reasonable.2.The results of case information analysis showed that the age of prepubertal boys with ASD was 5.08(4.50,5.75)years,height Z-score(HAZ)was 0.24(-0.44,0.53),weight Z-score(WAZ)was(0.33 ± 0.87),which was not significantly different from the age 5.17(4.58,5.54),HAZ 0.15(-0.07,0.57)and WAZ(0.12±0.84)of normal prepubertal boys(P>0.05).3.In ASD group,the ABC score was 54.00(37.00,74.00),SRS score was89.00(62.00,103.00),RBS-R score was 17.00(10.00,31.00)and CBCL score was 45.00(31.00,66.00).In normal controls,the ABC scale was 0.00(0.00,5.50),SRS scale score was 26.50(24.00,40.50),RBS-R scale was 3(1.00,9.75),and CBCL scale was 10.00(6.00,40.75).The Mann Whitney U test resulted in significantly higher scores for all scales in the ASD group than in the normal control group(P<0.001).4.The results of mass spectrometry analysis showed that the salivary cortisol concentration of boys with ASD was 864.04(552.71,1152.89)pg/ml,which was not significantly different from that of normal boys 798.70(622.26,1176.80)pg/ml;The level of dehydroepiandrosterone(DHEA)in saliva of boys with ASD was 71.99(19.92,166.03)pg/ml higher than that of normal boys20.33(6.96,77.70)pg/ml(P=0.004).The level of pregnenolone in saliva of boys with ASD was 29.95(12.03,168.06)pg/ml higher than that of normal boys 12.28(6.62,24.88)pg/ml(P=0.001).5.Spearman correlation analysis showed that the level of pregnenolone in saliva was correlated with the score of ABC scale(r=0.687,P<0.001).6.Logistic regression analysis showed that pregnenolone in saliva may be a potential biomarker of ASD(P=0.048).7.ROC analysis showed that the saliva pregnenolone had good distinguishing sensitivity between ASD group and normal control group(P<0.001).Conclusion:1.Prevalence of ASD in prepubertal boys independent of general demographic characteristics such as age and physical development;2.The CBCL,ABC,SRS,RBS-R scales provide good assessment of autistic behavioral symptoms in prepubertal boys with ASD;3.Boys with high salivary pregnenolone may be at greater risk for ASD than those with low pregnenolone.And that salivary pregnenolone has good discriminative sensitivity for ASD.Therefore,salivary pregnenolone may be a biomarker of ASD. |
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