<clinical_study>
  <!-- This xml conforms to an XML Schema at:
    https://clinicaltrials.gov/ct2/html/images/info/public.xsd -->
  <required_header>
    <download_date>ClinicalTrials.gov processed this data on September 20, 2023</download_date>
    <link_text>Link to the current ClinicalTrials.gov record.</link_text>
    <url>https://clinicaltrials.gov/ct2/show/NCT03018574</url>
  </required_header>
  <id_info>
    <org_study_id>KY20163381</org_study_id>
    <nct_id>NCT03018574</nct_id>
  </id_info>
  <brief_title>Candidate Gene Screening for Attention Deficit/Hyperactive Disorder (ADHD)</brief_title>
  <official_title>Candidate Gene Screening for 6-14 Year Old Patients With ADHD (Attention Deficit/ Hyperactivity Disorder)</official_title>
  <sponsors>
    <lead_sponsor>
      <agency>Xijing Hospital</agency>
      <agency_class>Other</agency_class>
    </lead_sponsor>
    <collaborator>
      <agency>Soochow University</agency>
      <agency_class>Other</agency_class>
    </collaborator>
  </sponsors>
  <source>Xijing Hospital</source>
  <oversight_info>
    <has_dmc>Yes</has_dmc>
  </oversight_info>
  <brief_summary>
    <textblock>
      Source: Sample bank of Xijing Hospital and Children's Hospital Affiliated to Soochow&#xD;
      University; Sample form: Whole blood; Estimated number of samples: 100 patients with ADHD and&#xD;
      age, sex matched healthy controls; Case exclusion criteria: all kinds of neuropsychiatric&#xD;
      disorders, IQ value of less than 70;&#xD;
&#xD;
      Study protocol:&#xD;
&#xD;
        1. Using Qiagen kit to extract the genomic DNA of 200 microliters of blood.&#xD;
&#xD;
        2. UV spectrophotometer test DNA purity of 260/280 close to 1.8 (1.8 ± 0.05), the&#xD;
           concentration of 100ng/μL or more before the next sequencing.&#xD;
&#xD;
        3. The extracted genomic DNA will be sent to Sangon Biology Engineering Limited Company&#xD;
           (Shanghai) and sequenced to find candidate mutations related to ADHD risk sequence.&#xD;
           According to NIH gene database, the longest transcript of NDRG2 (ID: 57447 gene,&#xD;
           https://www.ncbi.nlm.nih.gov/nuccore/NC_000014.9?&#xD;
           Report=genbank&amp;from=21016763&amp;to=21070872&amp;strand=true) (a total of 17 exons and 16&#xD;
           introns and the gene 5 'UTR and 3' UTR region) will be alignmented sequences to find&#xD;
           potential mutations.&#xD;
&#xD;
        4. Using the chi square analysis and other statistical methods to determine the&#xD;
           relationship between the mutations and susceptibility to ADHD.&#xD;
    </textblock>
  </brief_summary>
  <detailed_description>
    <textblock>
      Source: Sample bank of Xijing Hospital and Children's Hospital Affiliated to Soochow&#xD;
      University; Sample form: Whole blood; Estimated number of samples: 100 patients with ADHD and&#xD;
      age, sex matched healthy controls; Case exclusion criteria: all kinds of neuropsychiatric&#xD;
      disorders, IQ value of less than 70;&#xD;
&#xD;
      Study protocol:&#xD;
&#xD;
        1. Using Qiagen kit to extract the genomic DNA of 200 microliters of blood. (1) melt frozen&#xD;
           blood at room temperature; (2) take the blood of 0.2 ml into a sterile anti coagulation&#xD;
           centrifuge tube, add an equal volume of PBS phosphate buffer, after fully mixing&#xD;
           12000rpm centrifugation 5min, discard supernatant; (3) 66.7 L STE 2.4 L, 20% SDS, 37 DEG&#xD;
           C water bath 1h; (4) protease K plus 1 20mg/ml l mix, 55 DEG C water bath digestion&#xD;
           overnight (10 ~ 14h); (5) the digested samples treated with Tris saturated phenol, shake&#xD;
           well, 12000rpm centrifugal 10min; (6) the upper aqueous phase to a sterile centrifuge&#xD;
           tube; adding volume of Tris saturated phenol, shake well, 12000rpm centrifugal 10min;&#xD;
           (7) the upper aqueous phase was transferred to another sterile centrifuge tube, and an&#xD;
           equal volume of phenol was added: chloroform: isoamyl alcohol (25: 24: 1). The&#xD;
           oscillation of the vortex was fully mixed, and the 12000rpm was centrifuged to 10min;&#xD;
           (8) the upper aqueous phase was transferred to another sterile centrifuge tube, and an&#xD;
           equal volume of chloroform was added: iso amyl alcohol (24: 1), which was fully vibrated&#xD;
           and mixed with 12000rpm centrifugation 10min; (9) transfer the supernatant to another&#xD;
           sterile centrifuge tube; adding 2 times volume ethanol, the volume level of sodium&#xD;
           acetate 1/10 shake, until the flocculent precipitation of DNA visible; (10) the sample&#xD;
           is placed at -20 DEG C refrigerator freezer 30min or ice bath for 15 ~ 20min, after&#xD;
           removal of 12000rpm centrifugal 10min again, so that DNA precipitation; (11) the gun&#xD;
           head will pick DNA to another sterile centrifugal tube, with the amount of 70% ethanol&#xD;
           washing and shaking, to wash out impurities DNA; (12) out of ethanol, DNA by vacuum&#xD;
           drying or dry naturally, TE buffer adding dissolution, -20 stored at standby.&#xD;
&#xD;
        2. UV spectrophotometer test DNA purity of 260/280 close to 1.8 (1.8 ± 0.05), the&#xD;
           concentration of 100ng/μL or more before the next sequencing.&#xD;
&#xD;
        3. The extracted genomic DNA will be sent to Sangon Biology Engineering Limited Company&#xD;
           (Shanghai) and sequenced to find candidate mutations related to ADHD risk sequence.&#xD;
           According to NIH gene database, the longest transcript of NDRG2 (ID: 57447 gene,&#xD;
           https://www.ncbi.nlm.nih.gov/nuccore/NC_000014.9?&#xD;
           Report=genbank&amp;from=21016763&amp;to=21070872&amp;strand=true) (a total of 17 exons and 16&#xD;
           introns and the gene 5 'UTR and 3' UTR region) will be alignmented sequences to find&#xD;
           potential mutations.&#xD;
&#xD;
        4. Using the chi square analysis and other statistical methods to determine the&#xD;
           relationship between the mutations and susceptibility to ADHD.&#xD;
    </textblock>
  </detailed_description>
  <overall_status>Completed</overall_status>
  <start_date>May 2016</start_date>
  <completion_date type="Actual">January 2017</completion_date>
  <primary_completion_date type="Actual">January 2017</primary_completion_date>
  <study_type>Observational</study_type>
  <has_expanded_access>No</has_expanded_access>
  <study_design_info>
    <observational_model>Case-Control</observational_model>
    <time_perspective>Retrospective</time_perspective>
  </study_design_info>
  <primary_outcome>
    <measure>Gene point mutation in the genome of ADHD patients and healthy controls</measure>
    <time_frame>2016-2017</time_frame>
  </primary_outcome>
  <number_of_groups>2</number_of_groups>
  <enrollment type="Actual">100</enrollment>
  <condition>Gene Product Sequence Variation</condition>
  <arm_group>
    <arm_group_label>ADHD-patients</arm_group_label>
    <description>The whole blood sample from diagnoses of the children 6-14 years old with ADHD. Diagnoses of the children with ADHD were made in Xijing Hospital and Children's Hospital Affiliated to Soochow University according to criteria described in the Diagnostic and Statistical Manual of Mental Disorders (DSM-IV). Children with ADHD had an IQ score above 70.</description>
  </arm_group>
  <arm_group>
    <arm_group_label>Controls-healthy children</arm_group_label>
    <description>The whole blood sample from age- and gender- matched healthy 6-14 years old children</description>
  </arm_group>
  <biospec_retention>Samples With DNA</biospec_retention>
  <biospec_descr>
    <textblock>
      Whole blood&#xD;
    </textblock>
  </biospec_descr>
  <eligibility>
    <study_pop>
      <textblock>
        Diagnoses of the children with ADHD were made in Xijing Hospital and Children's Hospital&#xD;
        Affiliated to Soochow University according to criteria described in the Diagnostic and&#xD;
        Statistical Manual of Mental Disorders. Children with ADHD had an IQ score above 70.&#xD;
      </textblock>
    </study_pop>
    <sampling_method>Probability Sample</sampling_method>
    <criteria>
      <textblock>
        Inclusion Criteria:&#xD;
&#xD;
          -  Diagnoses of the children with ADHD were made in Xijing Hospital and Children's&#xD;
             Hospital Affiliated to Soochow University according to criteria described in the&#xD;
             Diagnostic and Statistical Manual of Mental Disorders (DSM-IV). Children with ADHD had&#xD;
             an IQ score above 70.&#xD;
&#xD;
        Exclusion Criteria:&#xD;
&#xD;
          -  Children who had a past history of or were currently affected by neurological&#xD;
             diseases, including convulsive disorders or brain damage; or who had any evidence of&#xD;
             comorbid psychiatric conditions, such as Tourette's syndrome, IQ below 70, pervasive&#xD;
             developmental disorder (autism), bipolar disorder, psychosis, language difficulties or&#xD;
             learning disorders (reading disorders, mathematics disorders and disorders of written&#xD;
             expression).&#xD;
      </textblock>
    </criteria>
    <gender>All</gender>
    <minimum_age>6 Years</minimum_age>
    <maximum_age>14 Years</maximum_age>
    <healthy_volunteers>Accepts Healthy Volunteers</healthy_volunteers>
  </eligibility>
  <overall_official>
    <last_name>Yan Li, PhD&amp;MD</last_name>
    <role>Principal Investigator</role>
    <affiliation>Xijing hospital of The fourth military medical university</affiliation>
  </overall_official>
  <location>
    <facility>
      <name>Xijing Hospital of the Fourth Military Medical University</name>
      <address>
        <city>Xi'an</city>
        <state>Shaanxi</state>
        <zip>710032</zip>
        <country>China</country>
      </address>
    </facility>
  </location>
  <location_countries>
    <country>China</country>
  </location_countries>
  <results_reference>
    <citation>Won H, Mah W, Kim E, Kim JW, Hahm EK, Kim MH, Cho S, Kim J, Jang H, Cho SC, Kim BN, Shin MS, Seo J, Jeong J, Choi SY, Kim D, Kang C, Kim E. GIT1 is associated with ADHD in humans and ADHD-like behaviors in mice. Nat Med. 2011 May;17(5):566-72. doi: 10.1038/nm.2330. Epub 2011 Apr 17.</citation>
    <PMID>21499268</PMID>
  </results_reference>
  <results_reference>
    <citation>Salatino-Oliveira A, Genro JP, Chazan R, Zeni C, Schmitz M, Polanczyk G, Roman T, Rohde LA, Hutz MH. Association study of GIT1 gene with attention-deficit hyperactivity disorder in Brazilian children and adolescents. Genes Brain Behav. 2012 Oct;11(7):864-8. doi: 10.1111/j.1601-183X.2012.00835.x. Epub 2012 Sep 7.</citation>
    <PMID>22897819</PMID>
  </results_reference>
  <results_reference>
    <citation>Vegt R, Bertoli-Avella AM, Tulen JH, de Graaf B, Verkerk AJ, Vervoort J, Twigt CM, Maat-Kievit A, van Tuijl R, van der Lijn M, Hengeveld MW, Oostra BA. Genome-wide linkage analysis in a Dutch multigenerational family with attention deficit hyperactivity disorder. Eur J Hum Genet. 2010 Feb;18(2):206-11. doi: 10.1038/ejhg.2009.148. Epub 2009 Aug 26.</citation>
    <PMID>19707245</PMID>
  </results_reference>
  <verification_date>January 2017</verification_date>
  <study_first_submitted>January 11, 2017</study_first_submitted>
  <study_first_submitted_qc>January 11, 2017</study_first_submitted_qc>
  <study_first_posted type="Estimate">January 12, 2017</study_first_posted>
  <last_update_submitted>February 22, 2018</last_update_submitted>
  <last_update_submitted_qc>February 22, 2018</last_update_submitted_qc>
  <last_update_posted type="Actual">February 26, 2018</last_update_posted>
  <responsible_party>
    <responsible_party_type>Sponsor</responsible_party_type>
  </responsible_party>
  <condition_browse>
    <!-- CAUTION:  The following MeSH terms are assigned with an imperfect algorithm            -->
    <mesh_term>Attention Deficit Disorder with Hyperactivity</mesh_term>
  </condition_browse>
  <patient_data>
    <sharing_ipd>Yes</sharing_ipd>
    <ipd_description>Publish all data in a paper</ipd_description>
  </patient_data>
  <!-- Results have not yet been posted for this study                                          -->
</clinical_study>


download date: ClinicalTrials.gov processed this data on September 20, 2023
link text: Link to the current ClinicalTrials.gov record.
url: https://clinicaltrials.gov/ct2/show/NCT03018574
org study id: KY20163381
nct id: NCT03018574
lead sponsor: 
collaborator: 
has dmc: Yes
Source: Sample bank of Xijing Hospital and Children's Hospital Affiliated to Soochow
      University; Sample form: Whole blood; Estimated number of samples: 100 patients with ADHD and
      age, sex matched healthy controls; Case exclusion criteria: all kinds of neuropsychiatric
      disorders, IQ value of less than 70;

      Study protocol:

        1. Using Qiagen kit to extract the genomic DNA of 200 microliters of blood.

        2. UV spectrophotometer test DNA purity of 260/280 close to 1.8 (1.8 ± 0.05), the
           concentration of 100ng/μL or more before the next sequencing.

        3. The extracted genomic DNA will be sent to Sangon Biology Engineering Limited Company
           (Shanghai) and sequenced to find candidate mutations related to ADHD risk sequence.
           According to NIH gene database, the longest transcript of NDRG2 (ID: 57447 gene,
           https://www.ncbi.nlm.nih.gov/nuccore/NC_000014.9?
           Report=genbank&from=21016763&to=21070872&strand=true) (a total of 17 exons and 16
           introns and the gene 5 'UTR and 3' UTR region) will be alignmented sequences to find
           potential mutations.

        4. Using the chi square analysis and other statistical methods to determine the
           relationship between the mutations and susceptibility to ADHD.
Source: Sample bank of Xijing Hospital and Children's Hospital Affiliated to Soochow
      University; Sample form: Whole blood; Estimated number of samples: 100 patients with ADHD and
      age, sex matched healthy controls; Case exclusion criteria: all kinds of neuropsychiatric
      disorders, IQ value of less than 70;

      Study protocol:

        1. Using Qiagen kit to extract the genomic DNA of 200 microliters of blood. (1) melt frozen
           blood at room temperature; (2) take the blood of 0.2 ml into a sterile anti coagulation
           centrifuge tube, add an equal volume of PBS phosphate buffer, after fully mixing
           12000rpm centrifugation 5min, discard supernatant; (3) 66.7 L STE 2.4 L, 20% SDS, 37 DEG
           C water bath 1h; (4) protease K plus 1 20mg/ml l mix, 55 DEG C water bath digestion
           overnight (10 ~ 14h); (5) the digested samples treated with Tris saturated phenol, shake
           well, 12000rpm centrifugal 10min; (6) the upper aqueous phase to a sterile centrifuge
           tube; adding volume of Tris saturated phenol, shake well, 12000rpm centrifugal 10min;
           (7) the upper aqueous phase was transferred to another sterile centrifuge tube, and an
           equal volume of phenol was added: chloroform: isoamyl alcohol (25: 24: 1). The
           oscillation of the vortex was fully mixed, and the 12000rpm was centrifuged to 10min;
           (8) the upper aqueous phase was transferred to another sterile centrifuge tube, and an
           equal volume of chloroform was added: iso amyl alcohol (24: 1), which was fully vibrated
           and mixed with 12000rpm centrifugation 10min; (9) transfer the supernatant to another
           sterile centrifuge tube; adding 2 times volume ethanol, the volume level of sodium
           acetate 1/10 shake, until the flocculent precipitation of DNA visible; (10) the sample
           is placed at -20 DEG C refrigerator freezer 30min or ice bath for 15 ~ 20min, after
           removal of 12000rpm centrifugal 10min again, so that DNA precipitation; (11) the gun
           head will pick DNA to another sterile centrifugal tube, with the amount of 70% ethanol
           washing and shaking, to wash out impurities DNA; (12) out of ethanol, DNA by vacuum
           drying or dry naturally, TE buffer adding dissolution, -20 stored at standby.

        2. UV spectrophotometer test DNA purity of 260/280 close to 1.8 (1.8 ± 0.05), the
           concentration of 100ng/μL or more before the next sequencing.

        3. The extracted genomic DNA will be sent to Sangon Biology Engineering Limited Company
           (Shanghai) and sequenced to find candidate mutations related to ADHD risk sequence.
           According to NIH gene database, the longest transcript of NDRG2 (ID: 57447 gene,
           https://www.ncbi.nlm.nih.gov/nuccore/NC_000014.9?
           Report=genbank&from=21016763&to=21070872&strand=true) (a total of 17 exons and 16
           introns and the gene 5 'UTR and 3' UTR region) will be alignmented sequences to find
           potential mutations.

        4. Using the chi square analysis and other statistical methods to determine the
           relationship between the mutations and susceptibility to ADHD.
observational model: Case-Control
time perspective: Retrospective
measure: Gene point mutation in the genome of ADHD patients and healthy controls
time frame: 2016-2017
arm group label: ADHD-patients
description: The whole blood sample from diagnoses of the children 6-14 years old with ADHD. Diagnoses of the children with ADHD were made in Xijing Hospital and Children's Hospital Affiliated to Soochow University according to criteria described in the Diagnostic and Statistical Manual of Mental Disorders (DSM-IV). Children with ADHD had an IQ score above 70.
arm group label: Controls-healthy children
description: The whole blood sample from age- and gender- matched healthy 6-14 years old children
Whole blood
study pop: 
sampling method: Probability Sample
criteria: 
gender: All
minimum age: 6 Years
maximum age: 14 Years
healthy volunteers: Accepts Healthy Volunteers
last name: Yan Li, PhD&MD
role: Principal Investigator
affiliation: Xijing hospital of The fourth military medical university
facility: 
country: China
citation: Won H, Mah W, Kim E, Kim JW, Hahm EK, Kim MH, Cho S, Kim J, Jang H, Cho SC, Kim BN, Shin MS, Seo J, Jeong J, Choi SY, Kim D, Kang C, Kim E. GIT1 is associated with ADHD in humans and ADHD-like behaviors in mice. Nat Med. 2011 May;17(5):566-72. doi: 10.1038/nm.2330. Epub 2011 Apr 17.
PMID: 21499268
citation: Salatino-Oliveira A, Genro JP, Chazan R, Zeni C, Schmitz M, Polanczyk G, Roman T, Rohde LA, Hutz MH. Association study of GIT1 gene with attention-deficit hyperactivity disorder in Brazilian children and adolescents. Genes Brain Behav. 2012 Oct;11(7):864-8. doi: 10.1111/j.1601-183X.2012.00835.x. Epub 2012 Sep 7.
PMID: 22897819
citation: Vegt R, Bertoli-Avella AM, Tulen JH, de Graaf B, Verkerk AJ, Vervoort J, Twigt CM, Maat-Kievit A, van Tuijl R, van der Lijn M, Hengeveld MW, Oostra BA. Genome-wide linkage analysis in a Dutch multigenerational family with attention deficit hyperactivity disorder. Eur J Hum Genet. 2010 Feb;18(2):206-11. doi: 10.1038/ejhg.2009.148. Epub 2009 Aug 26.
PMID: 19707245
responsible party type: Sponsor
mesh term: Attention Deficit Disorder with Hyperactivity
sharing ipd: Yes
ipd description: Publish all data in a paper