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血小板α 颗粒膜蛋白 (CD62P)和溶酶体膜蛋白 (CD63)是血小板活化标志物 ,但血小板膜蛋白极不稳定 ,易活化 ,造成实验结果不可靠 ,为此应采用统一标准的检测方法。为探讨该目的并了解糖尿病患者血小板的活化情况 ,实验中采用全血单克隆抗体直接免疫荧光标记法和流式细胞术 ,对正常人及 37例成人糖尿病患者进行了研究。正常人无固定组在标记后 30 ,60 ,90和 12 0分钟时 ,CD62P和CD63阳性率 (% )分别为 7.5 7± 2 .33,2 0 .5 0± 5 .70 ,2 8.70± 5 .67,36.5 2± 6.13及 0 .89± 0 .36,1.11± 0 .84 ,2 .35± 2 .0 2 ,5 .4 3± 3.66,其相应点的平均荧光强度各自为 1.5 7± 0 .13,1.88± 0 .0 8,2 .0 0±0 .0 9,2 .38± 0 .2 2及 3.91± 0 .11,4 .0 7± 0 .16,4 .38± 0 .14,4 .4 4± 0 .19,二项均随检测时间推移逐渐升高 ,其中阳性率增高更加显著。而正常人多聚甲醛固定组CD62P和CD63在相同各时间点的阳性率和平均荧光强度基本相同 ,未出现明显变化。此外 ,37例成人糖尿病患者CD62P和CD63的阳性率分别为 (14.11± 6.68) %及 (2 .71± 1.74 ) % ,比相对正常对照组显著升高 (P <0 .0 0 1,P <0 .0 5 )。由此可见 ,血小板CD62P和CD63非常敏感 ,在体外易受激活 ,采血后需立即标记 ,全部操作应在 30分钟内完成 ,
Platelet α-granule membrane protein (CD62P) and lysosomal membrane protein (CD63) are platelet activation markers, but platelet membrane protein is very unstable and easily activated, resulting in unreliable experimental results. Therefore, we should adopt a uniform standard detection method. To explore the purpose and to understand the activation of platelet in diabetic patients, normal human and 37 adult patients with diabetes were studied by direct immunofluorescent labeling with whole blood monoclonal antibody and flow cytometry. The positive rates (%) of CD62P and CD63 at 30, 60, 90 and 120 minutes after labeling were 7.5 7 ± 2 .33, 2.05 ± 5 .70 and 2 8.70 ± 5 .67, 36.5 ± 6.13 and 0.89 ± 0.36, 1 .1 ± 0 .84, 2 .35 ± 2 .0 2 and 5 .4 3 ± 3.66, respectively. The average fluorescence intensities of corresponding points were 1.5 7 ± 0 .13, 1.88 ± 0 .0 8,2 .0 0 ± 0 .0 9,2 .38 ± 0 .2 2 and 3.91 ± 0 .11, 4 .0 7 ± 0 .16, 4.38 ± 0 .14,4 .4 4 ± 0.19, both with the passage of time gradually increased, of which the positive rate increased more significantly. However, the positive rate and the average fluorescence intensity of CD62P and CD63 in normal paraformaldehyde fixation group at the same time point were basically the same, with no obvious change. In addition, the positive rates of CD62P and CD63 in 37 adults with diabetes were (14.11 ± 6.68)% and (2.71 ± 1.74)%, respectively, which were significantly higher than those in the control group (P <0.01, P < 0 .0 5). Thus, platelet CD62P and CD63 are very sensitive, susceptible to activation in vitro, need to immediately mark after blood collection, all operations should be completed within 30 minutes,