Data source
The data was from Zhejiang Ageing and Health Cohort Study, a community-based cohort study focusing on aging and health among the elderly in Zhejiang province, China [15]. It was conducted by Zhejiang Provincial Center for Disease Control and Prevention. Seven counties were randomly selected from a total of 90 counties in Zhejiang province. One town in each county and several communities in each town were then randomly selected. All permanent residents aged ≥60 years old in these selected communities were eligible and expected to be included in the study. In total, 13,955 individuals were eligible in the selected communities. The baseline survey was initiated in 2014, and 10,901 elderly were recruited and completed the survey (10,911 were primarily documented in the database, but 10 records were found duplicated later and were ultimately deleted). Three waves of follow-up surveys have been administered in 2015, 2016, and 2019–2020, especially. Due to funding restraint, the second wave of follow-up survey was administered in six counties, while other waves of follow-up surveys were administered in all seven counties.
A face-to-face interview based on a self-designed questionnaire was performed by trained research assistants for each participant at the baseline survey and each wave of the follow-up survey. The questionnaire included information on demographic characteristics, family status, reproductive history, medical disease, behavioral habits, diet habits, injury, depressive symptoms, self-care ability, and cognitive function. Data were checked by staff at Zhejiang Provincial Center for Disease Control and Prevention. Missing data and logical errors were fed back to the initial interviewers who would try to complete the dataset by reinvestigating the participants.
Selection of participants
Ten thousand nine hundred one participants were enrolled in the baseline survey of the Zhejiang Ageing and Health Cohort Study. Participants with depressive symptoms at baseline (n = 1107) as well as those with incomplete baseline items in the questions related to depressive symptoms (n = 7) and egg consumption (n = 4) were excluded from the dataset. Moreover, participants with the presence of cognitive impairment (n = 1173) at baseline were excluded due to the higher possibility of recall bias. It was examined through the mini-mental state examination (MMSE) in the cognitive function section within the questionnaire, with a lower score indicating a poorer cognitive function. The widely accepted cut-off point to define cognitive impairment in China (MMSE Chinese Standard) is education-specific [16]: 17/18 for people with lower than primary education level, 20/21 for people with primary education level, 24/25 for people with higher than primary education level. Finally, a cohort for the association between egg consumption and risk of depressive symptoms included 8610 participants.
Among them, 7575 (88.0%) completed the first wave of the follow-up survey. In the second wave of follow-up, 5791 (79.7%) from six counties completed the survey. 7203 (83.6%) completed the third wave of follow-up. Overall, more than half (4806, 55.8%) completed all three waves of the follow-up, 2668 (31.0%) completed two waves of the follow-up, 815 (9.5%) completed only one wave of the follow-up, and 321 (3.7%) never completed any wave of the follow-up. Hence, we included 8289 (95.8%) who attended the follow-up survey at least once in the final analysis (Fig. 1).
Assessment of depressive symptoms
Patient Health Questionnaire-9 scale (PHQ-9), a 9-question version of the Primary Care Evaluation of Mental Disorders, was adopted to evaluate depressive symptoms [17]. The total score for the nine items ranges from 0 to 27, with greater values indicating increased severity. Scores of 5, 10, 15, and 20 represented cut-off points for mild, moderate, moderately severe, and severe depressive symptoms, respectively. The PHQ-9 has been validated with acceptable psychometric properties for depression screening in the Chinese population [18]. Our purpose was to identify participants with mild to severe depressive symptoms in the elderly. Therefore, a cut-off score of five or more was used to define depressive symptoms in this study.
Egg consumption
Egg consumption at baseline was regarded as an exposure variable. Two items in the diet habits section of the baseline questionnaire were related to egg consumption: frequency of egg intake (days/week, “how many days did you have eggs every week generally in the last 12 months?”) and quantity of egg intake (eggs/day, “how many eggs did you have in the days you consumed eggs?”) [15].
Egg consumption (eggs/week) was calculated from frequency of egg intake multiplied by quantity of egg intake. We categorized participants into three groups according to their egg consumption: none or not weekly, < 3 eggs/week, and ≥ 3 eggs/week.
Covariates
Based on findings reported in the literature, variables from baseline survey described below were considered as potential confounders in our analysis: age (years, continuous variable), gender (“male” and “female”), race (“Han ethnicity” and “minority”), education level (“lower than primary”, “primary”, “junior middle”, “senior middle” and “college and above”), marital status (“single”, “married” and “divorced/widowed”), family income (“≤10,000”, “10,001 ~ 20,000”, “20,001 ~ 50,000”, “50,001 ~ 100,000” and “> 100,000” Chinese Yuan/year), body mass index (“< 18.5”, “18.5 ~ < 24” and “≥24” kg/m2), hypertension (“presence” and “absence”), diabetes (“presence” and “absence”), coronary heart disease (“presence” and “absence”), smoking (“never”, “past” and “current”), alcohol drinking (“never”, “past” and “current”), exercise (“yes” and “no”), tea drinking (“yes” and “no”), vegetable intake (“< 7” and “≥7” days/week), fruit intake (“< 3” and “≥3” days/week), red meat intake (“< 3” and “≥3” days/week), fish intake (“< 3” and “≥3” days/week). Detailed information on some of these variables was described as follows.
(1) Medical disease section of the questionnaire contained the items on the presence or absence of 16 common diseases, which are supposed to be formally diagnosed by a physician. Hypertension, diabetes, and coronary heart disease (CHD) were considered in this study. (2) The frequency of other food intake was also collected in the diet habits section of the questionnaire. Vegetables, fruits, red meat, and fish were included in this study. It was divided into two categories for each variable based on the median. (3) Han ethnicity is the major ethnicity in the region.
Statistical analysis
Descriptive statistics were applied to illustrate the general characteristics of included participants. The associations between general characteristics and egg consumption were examined by ANOVA, Kruskal-Wallis test, or Chi-square test as appropriate to variables. We used the log-binomial regression models for repeated measures with the Generalized Estimating Equations (GEE) method [19] to assess the longitudinal effect of egg consumption on the risk of depressive symptoms. Crude relative risks (RRs), 95% confidence intervals (CIs), and corresponding P values of depressive symptoms associated with egg consumption were calculated in model 1. In model 2, we adjusted for sociodemographic characteristics including age, gender, race, education level, marital status, and family income. In model 3, we additionally adjusted for BMI, hypertension, diabetes, coronary heart disease, smoking, alcohol drinking, exercise, and tea drinking. In model 4, vegetable intake, fruit intake, red meat intake, and fish intake were additionally adjusted. When the log-binomial model failed to converge, COPY method was adopted using a newly expanded data set that contained c-1 copies (c is usually set as 1000) of the original data and 1 copy of the original data with the dependent variable values interchanged [20]. Tests for linear trends were simultaneously implemented by assigning the median values to each category of egg consumption and modeling these values as a single continuous variable. Once the linear trend has been detected, the RR for each egg increment per week was calculated with the continuous variable of egg consumption (before transformation into the categorical variable) in the model. Furthermore, we also conducted stratified analyses to test whether observed associations varied by frequency of other food intakes. Meanwhile, interactions between egg consumption and these factors were checked through the addition of cross-product terms in the corresponding main-effects models. Finally, sensitivity analyses were adopted to evaluate the robustness of our results by 1) restricting the participants to those completing at least two waves of follow-up survey; 2) including the participants with depressive symptoms at baseline in GEE models, whose dependent variable value is expected to be similar (classified as depressive symptoms) at each wave of follow-up; 3) using the PHQ-9 score ≥ 9 to define depressive symptoms, which is generally accepted to screen for depression in China.
Statistical analyses were conducted using SAS Software Version 9.4 (SAS Institute Inc., Cary, NC, USA). A P value < 0.05 was considered statistically significant.