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ống sau đó được đặt trên vỏ bao băng, được lưu trữ trong các thùng chứa xốp, trở về phòng thí nghiệm của chúng tôi bằng chuyển phát nhanh qua đêm, ly tâm, và chia thành aliquots lưu trữ trong tủ lạnh nitơ lỏng ("130 ° C hoặc lạnh hơn). | inflammatory markers and the risk of coronary heart disease in men and women tubes were then placed on ice packs stored in Styrofoam containers returned to our laboratory by overnight courier centrifuged and divided into aliquots for storage in liquid-nitrogen freezers -130 C or colder . The levels ofC-reactive protein were determined by means ofa highly sensitive immunoturbidimetric assay with the use of reagents and calibrators from Denka Seiken this assay has a day-to-day variability of 1 to 2 percent. Levels ofsTNF-R1 sTNF-R2 and interleukin-6 were measured by means of enzyme-linked immunosorbent assays R D Systems which have a day-to-day variability of3.5 to 9.0 percent. Levels of inflammatory markers were largely unaffected by transport conditions and reproducible within subjects over time.22 23 Total high-density lipoprotein HDL and directly obtained low-density lipoprotein LDL cholesterol and triglycerides were measured according to standard methods with the use of reagents from Roche Diagnostics and Genzyme. Study samples were sent to the laboratory for analysis in randomly ordered batches and the laboratory personnel were unaware of a sample s case-control status. The study protocol was approved by the institutional review board of the Brigham and Women s Hospital and the Human Subjects Committee Review Board of Harvard School of Public Health. exclusions After the exclusion ofparticipants with missing data on biomarker levels our data sets consisted of 708 women 239 patients and 469 controls and 794 men 265 patients and 529 controls . The assay for interleukin-6 required slightly more plasma than we originally reserved for this assay among women. Therefore analyses involving interleukin-6 were restricted to the subgroup of 676 women for whom interleukin-6 levels were available. statistical analysis We analyzed the two cohorts separately. Inflammatory markers were divided into quintiles from the lowest to highest levels on the basis of the sex-specific .
