PDF《The association between BMP4 gene》

T (rs17563). Reactions were performed in a total volume of

20 μl. The thermocycling procedure consisted of initial denaturation at 95°C for

3 minutes,

35 cycles of denaturation at 94°C for

30 sec- onds, annealing at 60°C for

40 seconds, extension at 72°C for

1 minute and a final extension at 72°C for

10 minutes. The PCR products were analyzed by electrophoresis on 1% agarose gel [24]. Serum BMP4 and other cytokine Measurement Serum BMP4 was measured using the BMP4 Duoset enzyme linked immunoassay kit (R &

D Systems, Minneapolis, MN) according to manufacturer'

s instructions. Samples were assayed in duplicate. The interassay coefficient of variance was <

14% and the detection limit of the assay was 15.6 pg/mL [25]. Serum cystatin-C was measured using an immunoturbidimetric method (Roche Diagnostics, Indianapolis, IN) following the manufacturer'

s recommen- dations. The PIIINP levels were assayed using enzyme linked immunosorbent assay kits (Nivelles, Belgium). The Serum hs-CRP level was measured by using the high sensi- tivity CRP Vario assay (Abbott Laboratories........