| Objective:1.We investigated related social factors of child Attention deficit-hyperactivity disorder(ADHD)to regulate ADHD prevention and control measures.2.We studied varinance and related factors of Phthalate ester(PAEs)exposure to provide scientific basis for PAEs exposure prevention and control.3.We explored associations between PAEs exposure and child ADHD,combined Oppositional defiant disorder(ODD),related symptoms and behavior problems to provide potential clue for ADHD etiology research.4.We examined associations between neurotransmitter pathway gene polymorphism and ADHD,subtype and ADHD combined ODD to explore potential functional single nucleotide polymorphisms(FSNPs),and associations between PAEs exposure and ADHD among children with different genotype,and gene-PAEs exposure interaction to provide clue and evidence for study on ADHD genetics and potential mechanism of PAEs-ADHD.Methods:During December 2015 to May 2016,we enrollded diagnosed ADHD and health children in Liuzhou woman and children healthcare hospital and primary schools.After informed consent of parents and children,we collected ADHD related information,urinary samples and blood samples to conduct studies as follows:1.In this case-control study,social information of 233 ADHD children and 711 health children were investigated,then Odd ratio(OR)and 95%confidence interval(CI)of ADHD related factors were calculated by Logistic regression model.2.In this repeated measurements study,1299 urine samples of 461children were collected at different time points.Seven PAEs metabolites concentrations were measured by high performance liquid chromatography-mass spectrometry.Varinance and related factors of PAEs metabolites were analysed by mix linear models.3.In this 1:1 matched case-control study,ADHD related factors and PAEs concentrations of 225 ADHD children and health children were collected.Conditional and multivariate Logistic regressions were used to evaluate associations of PAEs exposure with ADHD and ADHD-ODD,respectively,and multiple linear regression was used to assess associations between PAEs concentrations and diseases symptoms and behavior problems.4.In this case-control study,blood samples of 168 ADHD and 233 health children were collected.Ten Single nucleotide polymorphisms(SNPs)of DAT,DRD4,NET and ADRA2A were tested by Miseq genotyping platform.Associations between SNPs and ADHD,subtype and ADHD-ODD,associations of Di 2-Ethyl Hexyl Phthalate(DEHP)exposure with ADHD among different genotype children,and gene-PAEs exposure interactions were analysed.Result:1.Male(OR=4.96,95%CI:3.28-7.49),6-9 years old(OR=1.77,95%CI:1.24-2.53),month income<3000 yuan per capita(OR=1.85,95%CI:1.31-2.61),preterm birth(OR=3.09,95%CI:1.15-8.33),maternal gernal/poor mood(OR=1.67,95%CI:1.18-2.36)and passive smoking(OR=1.81,95%CI:1.20-2.72)during pregnancy,gernal/poor parents relationship(OR=1.92,95%CI:1.31-2.82),father-child time<4 hours/week(OR=1.80,95%CI:1.16-2.79),hourse decoration last year(OR=1.62,95%CI:1.03-2.55)and child passive smoking(OR=1.77,95%CI:1.16-2.69)were related to ADHD.2.Mono-n-butyl phthalate(MBP)and Mono-2-ethyl-5-hydroxyhexyl phthalate(MEHHP)were the main urine PAEs metabolites,and the temporary reproducibility of MBP concentration were gernal(ICC=0.31),and the reproducibility of MEHHP,Mono-2-ethyl-5-oxohexyl phthalate(MEOHP)and Mono-2-ethylhexyl phthalate(MEHP)concentrations were poor(ICCs:0.21-0.29).In the mix linear models,MEHHP and MEOHP concentrations were related to 6-9 years old,male,minority and recent boltted drinks intake,and MEHHP concentration was also associated with recent plastic cups usage(all P<0.05).MBP concentration was related to urban,minority,frequent meat intake,rencent bottled drinks intake and shampoo usage(all P<0.05).3.Adjusted for covariates,the ORs of ADHD for children with the highest MEHHP,MEOHP,MEHP and Monobenzyl phthalate(MBzP)concentrations were 2.88(95%CI:1.37-6.02),2.77(95%CI:1.31-5.83),2.22(95%CI:1.12-4.39)and 2.14(95%CI:1.12-4.09),respectively,compared to the lowest exposure children.Besides,MEHHP,MEOHP and MEHP concentartions were associated with inattention scores,hyperactivity/impulse scores and oppositional defiant scores with positive dose-dependent relationship(Ptrend<0.05).MEHHP,MEOHP and MEHP concentartions were also related to ADHD-ODD,and the ORs were 2.05(95%CI:1.21-3.50),2.17(95%CI:1.25-3.76)and 1.98(95%CI:1.15-3.41),respectively.Regrading to children behavior checklists,MEHHP,MEOHP and MEHP concentartions were associated with inattention problems(β1=0.64,95%CI:0.04-1.23;P2=0.60,95%CI:0.01.19;β3=0.87,95%CI:0.18-1.56),aggressive behaviors(β1＝1.41,95%CI:0.38-2.44;β2=1.37,95%CI:0.35-2.38;β3=1.50,95%CI:0.30-2.70)and externalization behavior(β1=1.74;95%CI:0.35,3.16;β2=1.72,95%CI:0.34-3.09;β3=1.80,95%CI:0.17-3.43).There were positive associations between MEHP concentration and depression behavior(β=0.70,95%CI:0.05-1.34)and interlization behaviors(β=1.32,95%CI:0.06-2.58).In addition,MBzP concentration were associated with girls’ withdrawal behavior(β=0.69,95%CI:0.02-1.35)with sex difference(Psex=0.011).4.Adjusted for sex and age,the ORs of ADHD and ADHD-ODD for children with DRD4 rs747302 GG genotype were 2.46(95%CI:1.43-4.23)and 3.69(95%CI:1.23-11.09)times than CC children,respectively.The OR of ADHD for children with DRD4 rs3758653 CC genotype was 0.09(95%CI:0.01-0.73)times than TT children.The OR of ADHD-Inattention for DRD4 rs 1800955 CT children was 1.84(95%CI:1.05-3.21)times than CC children.The ORs of ADHD and ADHD-ODD for children with ADRA2A rs521674 AT genotype were 1.87(95%CI:1.23-2.84)and 3.16(95%CI:1.46-6.83)times than TT children.The OR of ADHD-Inattention for children with ADRA2A rs2484516 CG was 0.44(95%CI:0.22-0.89)times than CC children.The OR of ADHD for children with NET rs40615 AT-AA genotype was 0.64(95%CI:0.43-0.97)times than TT children,and the OR of ADHD-ODD for children with A allele was 0.49(95%CI:0.25-0.95)times than children with T allele.The OR of ADHD for children with NET rs 168924 G allele was 1.66(95%CI:1.15-2.39)times than children with A allele.The OR of ADHD-ODD for children with NET rs28386840 AT-TT genotype was 2.28(95%CI:1.05-4.96)times than AA children.Adjusted for covariates,with the increasing of MEHHP and MEOHP concentrations,the ORs of ADHD for children with DRD4 rs747302 CG-GG genotype were 1.73(95%CI:1.10-2.73)and 1.92(95%CI:1.20-3.09),respectively.With the increasing of MEHHP,MEOHP and MEHP concentrations,the ORs of ADHD for children with DRD4 rs1800955 CT-CC genotype were 1.76(95%CI:1.03-3.01),1.91(95%CI:1.11-3.30)and 1.71(95%CI:1.04-2.82),respectively;and the ORs of ADHD for children with NET rs168924 GG genotype were 1.67(95%CI:1.02-2.72),1.67(95%CI:1.10-3.03)and 1.67(95%CI:1.09-2.99),respectively.There were no significant interactions of gene-DEHP(Pinteraction>0.05).Conclusions:1.Boys aged 6-9 years old and preterm children should be screened preferentially for ADHD in order to find susceptible children as early as possible.Effects of adverse factors during pregnancy,poor parent’s and parent-child relationship,and adverse family environment on children ADHD are recommended to health education.2.Children expose to PAEs commonly with relatively high exposure level.The varinance of temporaty PAEs exposure indicate the necessity of repeated measurement to assess exposure more accurately.PAEs exposure were related to food-intake behavior,especially for recent bottled drinks intake and plastic cups use.3.High DEHP exposure are related to ADHD,ADHD-ODD and disease symptoms with dose-dependent relationship,suggesting that DEHP may contribute to the etiology of ADHD or disease progression.4.The polymorphisms of DRD4(rs747302,rs3758653,rs1800955),ADRA2A(rs521674,rs2484516)and NET(rs 168924,rs40615,rs28386840)are associated with ADHD,subtype and ADHD-ODD,providing clue for further study on potencial FSNP related to ADHD.5.Among children with DRD4 rs747302 CG-GG genotype,DRD4 rs1800955 CT-CC genotype and NET rs168924 GG genotype,DEHP exposure are positively related to ADHD,indicating that the potencial mechanism on the associations between DEHP and ADHD may involve those SNPs,and further researches are needed.Innovations:In the study,we combined macroscopic epidemiology,environmental hygiene and molecular epidemiology research to comprehensively investigate ADHD risk factors and identify ADHD etiology from sociological field,environmental field,genetic field,to provide clue and evidence for ADHD prevention and control measures and further study on combined diseases.Besides,the ORs of DEHP exposure for children ADHD and ADHD-ODD are identified,which fill a gap in the field.We also recognized the associations between DEHP and ADHD among children with different genotype,which provide some clues for potential mechanism.In addition,we observed the relationship between PAEs and external exposure factors by repeated measurements among children,which also fill a gap in the field. |
