Effects of Tobacco Smoke Exposure on Epigenetic Mechanism and Immunological Development

• Change from Baseline in cotinine level at 3 months [ Time Frame: 3 month ] [ Designated as safety issue: Yes ]
  Cotinine was the major proximate metabolite of nicotine,has been widely used as a biomarker of tobacco exposure. We will detect the urinary cotinine for children who had active smoking habit.
  
  
• Change from Baseline in smoking status at 3 months [ Time Frame: 3 months ] [ Designated as safety issue: Yes ]
  The smoking staus for children will be measure by structure questionnaire.
  
  

• Change from Baseline in DNA methylation at 3 months [ Time Frame: 3 months ] [ Designated as safety issue: Yes ]
  
• Change from Baseline in immunological cytokine at 3 months [ Time Frame: 3 months ] [ Designated as safety issue: Yes ]
  
• Change from Baseline in miRNA expression at 3 months [ Time Frame: 3 months ] [ Designated as safety issue: Yes ]
  
• Change from Baseline in immunological function at 3 months [ Time Frame: 3 months ] [ Designated as safety issue: Yes ]
  

Behavioral: Tobacco smoke reduction trial intervention program
Both the parents and children will receive 8 times intervention program weekly in first session. The telephone counseling will reduce the child's tobacco smoke exposure every weekly in second session. The interventionist and child jointly select behavioral goals for reducing tobacco smoke exposure for the child to work toward. The goals will be formalized as a written smoke exposure reduction plan for any of four specific behaviors, as relevant to that family: (a) smoking cessation for the child, (b) making the child's primary home smoke-free, (c) making non-home locations smoke-free.

Teenager Smoking Reduction Trial (TSRT)

1. Study subjects

   TSRT is a solution-oriented trial aiming to evaluate the effectiveness of tobacco smoke reduction among teenagers. Eligible teenagers will fit the following criteria: (a) age between 12 and 16 years old; (b) have active smoke habits in past 6 months; (c) amount of smoke exceed 10 cigarettes per week. The subjects will be recruited from our previous database for schoolchildren. Telephone interview will be used to screen the eligibility for the teenagers. Recruitment will occur over 3-month period from 2011 April to June. The procedure for TSRT is showed in figure 2. In this plan, we will recruit at least 31 asthmatic children and 31 non-asthmatic children who have active smoke habits.

2. Data collection

A baseline data and at the 1-month and 2-month follow-up visits, the primary caregiver provided data by personal interview on parents reports. The demographic information in questionnaires included age, gender, level of education.

Teenagers will report the frequency of cigarette smoking by using a structured questionnaire. The definition of tobacco smoke exposure including: (a) ''Do you currently smoke?''-yes/no; (b) ''When did you begin to smoke regularly?''; (c) Smokers were also asked for the number of cigarettes they smoked on an average day. Nicotine dependence will be examined by using structure questionnaires from Fagerstrom model.

We will contact eligible teenagers individually to get informed consent before data collection. The effects of our teenager smoking reduction intervention program will be assessed by primary outcomes such as cotinine/creatinine ratio (CCR) and the status of tobacco smoke exposure from questionnaire. No matter which group the participant belongs to, intervention and usual control group, the secondary outcomes in all participants will be tested as follows:

1. Cytokines. We will test levels of interleukin 4, 5, 13 (IL-4, 5, 13), tumor necrosis factor-α (TNF-α), interferon-gamma (INF-γ) and toll-like receptors 2, 4, 9 (TLR-2, 4, 9).
2. Immunotoxicological reaction. We will determine levels of exhaled nitric oxide (eNO), serum eosinophil cationic protein (ECP) and absolute eosinophil count (AEC).
3. Oxidative stress markers. Levels of the markers 8-hydroxy-2-deoxyguanosine (8-OHdG), malondialdehyde (MDA) and F2-isoprostane (Iso-P) will be examined.

After first session is applied, all participants will be tested for primary outcomes and secondary outcomes at baseline, month 1 and month 3 of the intervention process. We can then compare the biomarkers before and after the intervention program.