Study participants and exclusion criteria
Relevant clinical and sociodemographic data were obtained from Kangbuk Samsung Health Study (KSHS). KSHS is a cohort study to investigate the medical data of Koreans who have received medical health check-up in Kangbuk Samsung Hospital. Korea’s Industrial Safety and Health law orders that all of Korean employees should receive medical health check-up annually or biennially. Ethics approvals for the study protocol and analysis of the data were obtained from the institutional review board (IRB) of Kangbuk Samsung Hospital (IRB No. KBSMC 2020-09-25). All procedures performed in studies involving human participants were in accordance with the ethical standards of the IRB of the Kangbuk Samsung Hospital and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. IRB of Kangbuk Samsung Hospital approved the exemption of informed consent for the study because we only assessed retrospective data with de-identified personal information obtained from routine health check-up.
Among study participants in KSHS, we initially enrolled 136,405 subjects who had responded to semi-quantitative food frequency questionnaire (FFQ) including mushrooms consumption and Center for Epidemiologic Studies Depression (CES-D) between March 2011 and December 2012. Among these subjects, we excluded 820 subjects with taking sedative or anxiolytic medications and 15,148 subjects with depressive symptom in baseline analysis. Furthermore, 14,541 subjects with missing value in covariate data (e.g. BMI, hypertension, education) and 3,340 subjects with a history of serious medical diseases (e.g. coronary heart disease, stroke, and cancer) were further excluded. Out of the remaining 102,556 subjects, we finally recruited 87,822 participants who revisited and responded to the CES-D questionnaire from January 2013 to December 2018. Therefore, the total number of eligible study participants was 87,822.
Clinical and sociodemographic data
Study data include medical history assessed by self-administered questionnaire, anthropometric measurements and laboratory measurements. All study subjects were asked to respond to a health-related behavior questionnaire, which included the topics of alcohol consumption, smoking and exercise. Marriage, education, income, and occupation, which may influence psychological state and dietary patterns, were also included in the questionnaire. Hypertension was defined as a prior diagnosis of hypertension or having a measured BP ≥140/90 mmHg at initial and follow up examinations. Trained nurses measured BP on sitting position by automatic device (53000-E2, Welch Allyn, USA) three times after a 5 min rest with at least 30 seconds interval. Final BP levels were obtained as average of second and third BP measurements. The BMI was calculated by dividing weight (kilograms) by square of height (meters2). DM was defined as one of following conditions; fasting glucose ≥ 126 mg/dL, hemoglobin A1 c (HbA1c) ≥ 6.5%, and a prior diagnosis of DM (29).
Blood samples were collected after more than 12 hours of fasting and were drawn from an antecubital vein. The fasting serum glucose was measured using the hexokinase method, and hemoglobin A1c (Hba1c) was measured using an immunoturbidimetric assay with a Cobra Integra 800 automatic analyzer (Roche Diagnostics, Basel, Switzerland). Serum uric acid levels were measured enzymatically using an automatic analyzer Advia 1650 Autoanalyzer, Bayer Diagnostics; Leverkusen, Germany).
Assessment of FFQ data
We assessed the dietary intake of KSHS participants using the FFQ that was developed for the Korean genome epidemiologic study. The dietary data to design the FFQ were obtained from the Korea Health and Nutrition Examination Survey (30, 31). A detailed description of the FFQ (30) and its validation in the Korean population has been described in a previous study (31). The frequency of food consumption was composed of nine categories (e.g., mushroom intake was categorized never or rarely, once a month, two or three times a month, once or twice a week, three or four times a week, five or six times a week, one times a day, two times a day, and more than three times a day) and three serving sizes for each food. In mushrooms consumption, one serving size in mushrooms consumption was 30g and the serving size was classified into 15g, 30g, and 45g per day.
Participants answered their intake of oyster mushrooms and other mushrooms were categorized into five group according to mushroom consumption as follows: rare/never, <1/month (<one serving size/month), 1/month-1/week (one serving size/month ≤ ~ < one serving size/week), 1-3/week (one serving size/week ≤ ~ < three serving sizes/week), ≥ 3/week (≥ three serving sizes/week). Total energy and nutrient intake was calculated by the Can-Pro 3.0 software developed by The Korean Nutrition Society (32).
Assessment of depressive symptom
Depressive symptoms were assessed using the Korean versions of CES-D scale (33). The CES-D is a self-report questionnaire designed to assess the current presence of depressive symptoms in the general population (34). We used the 4-factors 20-items CES-D Scale with scores ranging from 0 to 3, with 0 indicating that the depressive symptom was experienced rarely and 3 indicating that the symptom was experienced most of the time in the past week. (e.g. “I thought my life had been a failure.” 0=seldom (not at all or less than 1 day), 1= sometimes (1~2 days), 2 = often (3~4 days), 3 = almost always (5~7 days)). This scale has been widely used across the world and shown the validity and reliability in the Korean general population (33). Depressive symptom was defined in the total score of CES-D ≥16. Therefore, in baseline analysis, subjects with CES-D ≥16 were regarded as the presence of depressive symptom in baseline and excluded from study participants. During follow-up, the subjects who newly fulfilled CES-D ≥16 were determined as the cases of incident depressive symptom.
STATISTICAL ANALYSES
The baseline parameters among groups of mushrooms consumption are presented as means ± standard deviation for continuous variables and as proportions for categorical variables. Main clinical characteristics and parameters among study groups were compared using ANOVA for continuous variables and chi-square test for categorical variables.
A Cox proportional hazards model was used to calculate the unadjusted and multivariable-adjusted hazard ratio (HR) and 95% confidence intervals (CI) for depressive symptom (multivariable adjusted HR [95% CI]) in each study group. The models were adjusted for multiple covariates including age, BMI, sex, alcohol intake, hypertension, diabetes, smoking, marital status, education, and total calorie intake. The covariates of the multivariable model were selected for the presence of significant differences between groups or established risk factors for depression. The incidence cases, incidence density (incidence cases per 1,000 person-years), person years of each group were calculated. Trend analysis conducted with median value of mushroom consumption. Given the different prevalence of depression in men and women, gender subgroup analyses were conducted. Furthermore, analyses by age subgroup were also conducted in consideration of different food preferences for each age subgroup (< 40 or ≥ 40 years old).
All statistical analyses were performed using R 3.6.3 (R Foundation for Statistical Computing, Vienna, Austria), and a value of P < 0.05 (two-sided) was considered statistically significant in all analyses.