Cui Wei, Ma Wenxin, Han Yehua, Lin Qisui
Department of Clinical Laboratory, Peking Union Medical College Hospital, CAMS & PUMC, Beijing 100730.
Chin Med Sci J. 2003 Mar;18(1):26-30.
To establish a flow cytometric internal standard method for counting platelet-derived microparticles (PMPs) and to study its clinical significance.
PMPs suspension (platelet poor plasma, PPP) was extracted by gradual centrifugation. According to the size of PMPs, 3 microm and 0.8 microm latex beads were used as internal standards for the quantitation. PMPs were counted by adjusting flow cytometric discrimination and voltage of forward scatter and side scatter.
In 30 healthy donors, the average concentration of resting PMPs was (1.2 x 10(5) +/- 5.7 x 10(4))/ml and that of activated PMPs was (1.6 x 10(6) +/- 9.1 x 10(5))/ml. Compared with healthy donors, PMPs mean value was significantly higher (P < 0.001) in 18 patients with coronary artery disease, 12 with acute cerebral infraction and 23 with chronic renal failure [the average PMPs concentration, (6.1 x 10(5) +/- 2.5 x 10(5))/ml, (6.8 x 10(5) +/- 3.4 x 10(5))/ml and (5.9 x 10(5) +/- 3.1 x 10(5))/ml respectively]. However, no significant difference in PMPs concentration was observed in 25 patients with acute leukemia and severe thrombocytopenia during the aplastic phase after chemotherapy [(1.3 x 10(5) +/- 6.1 x 10(4))/ml, (P > 0.05)].
PMPs is a useful indicator in monitoring platelet activation, and plays an important role in thrombotic disease. By flow cytometric internal standard method, PMPs can be counted rapidly and accurately, which may be very helpful in interlaboratory comparative studies.
建立流式细胞术内标法计数血小板衍生微粒（PMPs）并研究其临床意义。
通过逐步离心提取PMPs悬液（乏血小板血浆，PPP）。根据PMPs大小，使用3微米和0.8微米的乳胶微球作为定量内标。通过调整流式细胞术的鉴别参数以及前向散射和侧向散射电压来计数PMPs。
30名健康供者中，静息PMPs的平均浓度为（1.2×10⁵±5.7×10⁴）/毫升，活化PMPs的平均浓度为（1.6×10⁶±9.1×10⁵）/毫升。与健康供者相比，18例冠心病患者、12例急性脑梗死患者和23例慢性肾衰竭患者的PMPs平均值显著更高（P<0.001）[平均PMPs浓度分别为（6.1×10⁵±2.5×10⁵）/毫升、（6.8×10⁵±3.4×10⁵）/毫升和（5.9×10⁵±3.1×10⁵）/毫升]。然而，25例急性白血病患者化疗后再生障碍期严重血小板减少时，PMPs浓度无显著差异[（1.3×10⁵±6.1×10⁴）/毫升，（P>0.05）]。
PMPs是监测血小板活化的有用指标，在血栓性疾病中起重要作用。通过流式细胞术内标法可快速准确地计数PMPs，这可能对实验室间的比较研究非常有帮助。
