Study population
The CKB is a prospective cohort study of over 0.5 million adults recruited from ten diverse areas (five rural and five urban) in China, selected to cover a wide range of risk exposures, disease patterns and stages of economic development. Details of the study design, methods and population have been previously reported [20]. In brief, between June 2004 and July 2008, all permanent residents (aged 35–74 years; not severely disabled) in pre-selected communities or villages were invited to participate in the study. Among them, about one in three (33% in rural and 27% in urban areas) responded. A total of 512,713 participants (including a few who were just outside the target age range) were included in our baseline database. All participants provided written informed consent. Regional, national and international ethics approval was obtained prior to the start of recruitment.
For the current study, we excluded participants with baseline prevalent diabetes (n = 30,299), ischaemic heart disease (IHD; n = 15,472), stroke or transient ischaemic attack (n = 8884), or cancer (n = 2577) or those with missing values for BMI (n = 2), leaving 461,036 participants in the analysis (please note that some participants were excluded for meeting more than one criteria).
Baseline data collection in CKB
Information on sociodemographic status, smoking [21], alcohol drinking, physical activity [22], medical history and diet [23, 24] were collected by trained health professionals using a laptop-based questionnaire. Each participant provided a 10 ml venous blood sample (with time since last eating or drinking any energy-containing food or beverage recorded). Anthropometry (e.g. body weight, height, waist circumference) [3] and BP [25] were measured following standard protocols. BMI was calculated as weight (kg) divided by height squared (m2). In addition, body fat percentage (BF%) was estimated using a TBF-300 monitor (Tanita, Tokyo, Japan). Random blood glucose levels were measured immediately following sample collection using the SureStep Plus System (Johnson & Johnson, New Brunswick, NJ, USA), which provided plasma-equivalent readings and was regularly calibrated with manufacturer’s control solutions. Individuals who did not report a history of diabetes but who had random blood glucose ≥11.1 mmol/l or fasting blood glucose ≥7.0 mmol/l were defined as having screen-detected diabetes. Participants with either screen-detected diabetes or self-reported prior history of physician-diagnosed diabetes were classified as prevalent diabetes and excluded from the present study.
Dietary assessment
Information on consumption frequency (daily, 4–6 days/week, 1–3 days/week, monthly or never/rarely) of red meat (fresh and processed pork, beef and lamb/mutton), poultry (chicken, duck and goose) and fish (fish and shellfish) was collected using a validated interviewer-administered laptop-based questionnaire asking participants to report their eating habits during the past 12 months. The questionnaire has good reproducibility and relative validity against multiple 24 h recalls (weighted κ was 0.60, 0.61 and 0.75, respectively, for red meat, poultry and fish intake) [26]. In addition, strong positive associations were found between red meat consumption and blood levels of creatinine, total choline and sphingomyelin, and between fish consumption and blood levels of docosahexaenoic acid (DHA), DHA/fatty acid ratio, and total n-3 fatty acids (see electronic supplementary material [ESM] Fig. 1).
Following the completion of the baseline survey (2004–2008), 5–6% of the surviving participants were randomly selected to participate in re-surveys in order to understand the long-term variations and measurement errors of various baseline exposures. During the re-survey conducted in 2013–2014 (response rate 76%), the quantity of each food group consumed in addition to the consumption frequency was recorded, allowing us to estimate the usual mean amount consumed (i.e. average intake level during follow-up period) for each baseline exposure category.
Follow-up for incident diabetes
The vital status of each participant was obtained periodically through China’s Disease Surveillance Points (DSP) system [27] (death registry checked annually against local residential and health insurance records, and by street committees or village administrators). In addition, information on diabetes incidence was collected through linkages with chronic disease registries (for IHD, stroke, cancer and diabetes) and national health insurance claim databases, which provided almost universal (~99%) coverage of all hospitalisations for participants in the study. Both fatal and non-fatal events were coded using ICD-10 (https://icd.who.int/browse10/2014/en) by staff who were blinded to baseline information [20]. For the present study, incident diabetes included all recorded cases (E10-E14) that occurred between the ages of 35 and 79 years. A medical record review of approximately 1000 incidences of diabetes confirmed the validity of diabetes diagnosis (positive predictive value 97%). By 1 January 2017 (global censoring date), only 5276 (~1%) participants were lost to follow-up and they were censored in the prospective analyses.
Statistical analysis
To ensure an adequate number of diabetes cases in each consumption category for the prospective analyses, individuals were classified into four groups for red meat (daily, 4–6 days/week, 1–3 days/week and <1 day/week) and fish (≥4 days/week, 1–3 days/week monthly and never/rarely) consumption, and three groups for poultry consumption (weekly, monthly and never/rarely) by combining those original categories with less than 5% participants into the adjacent categories.
Means (SDs) or percentages of baseline characteristics were calculated across categories of each dietary exposure, adjusting for age, sex and region, where appropriate, using either multiple linear regression for continuous outcomes or logistic regression for binary outcomes. Cross-sectional associations of each dietary exposure under study with adiposity (BMI, waist circumference and BF%) were examined in men and women separately using multiple linear regression analyses. Adjustments were made for age (continuous variable), region (ten regions), smoking (four categories), alcohol intake (four categories), education (four categories), income (four categories), physical activity (continuous variable) and fresh fruit intake (five categories), and mutual adjustment for intake of the other two exposure variables. Analyses for waist circumference and BF% were additionally adjusted for BMI.
HRs and 95% CIs for diabetes incidence across exposure categories were estimated using Cox proportional hazards models, stratified by age-at-risk (groups of 5 years), sex and region, and adjusted for potential confounders including the above-mentioned covariates and family history of diabetes (dichotomous). Except for fresh fruit [24] and the three dietary exposures under study, no other dietary variables were included in the main models because none were associated with diabetes risk in the current analysis. In model 4, BMI (continuous) was also added in as a covariate. The proportion of diabetes risk explained by BMI was calculated as follows: [(loge HRmodel3 − loge HRmodel4)/loge HRmodel3] × 100%. The mean proportion and associated 95% CIs were obtained through bootstrap techniques with 1000 replications. The ‘floating absolute risk’ method was used to calculate 95% CIs of HRs in all exposure categories (including the reference category), without altering the point estimates. This method allows valid comparisons to be made between any two exposure groups for polychotomous risk factors [28]. We used data from 20,084 participants who attended the re-survey in 2013–2014 to correct for regression dilution bias [29, 30] and quantify the mean usual consumption quantities for each baseline exposure category (ESM Methods). The HR for each 50 g/day of usual red meat, poultry and fish intake was calculated using Cox regression analyses.
Stratified analyses by potential effect modifiers (e.g. sex, region, socioeconomic status [SES] and BMI) were performed and χ2 tests for trend and heterogeneity were applied to the loge HR and its SE. Comparison of HRs for the first and second halves of the follow-up period revealed no clear evidence of departure from the proportional hazards assumption. Sensitivity analyses were performed by excluding the first 2 years of follow-up or participants with incident cardiovascular disease (CVD) and cancer during follow-up, and by additional adjustment for other dietary factors and other adiposity indices.
All analyses were conducted using SAS (version 9.3, SAS Institute, Cary, NC, USA). Graphs were plotted using R 3.3.2 (https://www.R-project.org/).