Balkan Journal of Medical Genetics

CHRONIC OBSTRUCTIVE PULMONARY DISEASE RISK AND SMOKING CESSATION CHANGES INDUCED BY CHRNA5-A3 AND CHRNB3-A6 VARIATION IN A CHINESE MALE POPULATION
Zhao L1,, Zou L-Y2,, Cheng B-F3, Yu X-J4, Zou J-H5,*, Han W6,*
*Corresponding Author: Dr. Wei Han, Department of Pulmonary Medicine, Qingdao Municipal Hospital, Qingdao University, No. 5 Donghai Mid Road, Qingdao, People’s Republic of China. Tel: +86-185- 6185-7838. E-mail: sallyhan1@163.com. And: Dr. Jian-Hong Zou, Center of Diseases Control of Qingdao Shi-Bei District, No. 3 Deping Road, Qingdao, People’s Republic of China. Tel: +86-185-6185-7599. E-mail: 277517366@qq.com -Long Zhao, Ling-Yan Zou contributed equally to this study.
page: 51

MATERIALS AND METHODS

Research Participants. From January 2017 to May 2018, we recruited patients admitted to the Second Ward of Respiratory Department of Qingdao Municipal Hospital, Qingdao, Shandong Province, People’s Republic of China (PRC). The study consisting of 823 male smokers with COPD (407 patients with COPD who failed to quit smoking and 416 non smokers with COPD ≥1 year) and 435 healthy male smokers as control subjects. All subjects were recruited from the Smoking Cessation Clinic and the Medical Examination Department of Qingdao Municipal Hospital. Demographic information, including age, body mass index (BMI), and detailed circumstances of smoking, was collected in interviews conducted by trained medical doctors. Those smoking at least 20 packs in their life, or smoking at least one cigarette a day for more than 1 year were classified a smokers, non smokers were classified as having quit smoking ≥1 year. Smoking COPD subjects meet the following criteria: for those who still smoke or have quit smoking, physician-diagnosed COPD, pulmonary function test showing post-bronchodilator forced expiratory volume (FEV) in 1 second (FEV1)/forced vital capacity (FVC) of less than 70.0% and FEV1 of less than 80.0% predicted in the Global Initiative for COPD 2015 (GOLD 2015; https://glodcopd/org/). Subject exclusion criteria: definite diagnosis of lung cancer, asthma or smoking-related cancer. Cases were matched according to age, BMI and smoking history. The study was approved by Qingdao Municipal Hospital Ethics Committee (approval number: KYLL2010058). All patients participating in the program signed an informed consent form. Analysis. Blood samples of all participants were collected. Using the DNA extraction kit (Tiangen Biotech Co. Ltd., Beijing, PRC) to extract the genomic DNA of the subjects. Genotyping was carried out commercially using Sequenom MassARRAY® via Beijing Genomic Institute (BGI) (Shenzhen, PRC) [13]. In order to ensure the repeatability and consistency of the test, we randomly selected 5.0% of the samples for a second test. In addition, in order to further verify the accuracy of the BGI results, we also genotyped some samples by direct sequencing or restriction enzyme digestion. Statistical Analysis. The t-test was used to confirm the difference in population characteristics and gene distribution between the experimental group and the control group. The χ2 test was used to compare the demographic characteristics of the case group and the control group. The correlation between each SNP and the risk of smoking and nicotine-dependence in COPD patients was evaluated by a logistic regression model, which adjusted the age, BMI, current smoking status and the number of packages per year for long-term smokers. Age, BMI, current smoking status and pack-years of the number of cigarettes smoked per year, including those who quit and those who are currently smoking, were analyzed in a hierarchical manner to assess whether there was any difference between these subgroups. All statistical analyses adopted a two-sided test, and a p value of <0.05 was considered to be statistically significant. The Statistical Package for the Social Sciences (SPSS) version 22 (IBM® SPSS; www.ibm.com/SPSSStatistics/) was used for all statistical analysis.

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