目的分析一先天性血小板减少症家系的临床、实验室特点,并探讨其分子发病机制。方法收集该家系成员的临床资料,采集先证者及其家系成员的静脉血,分别进行全自动及人工血小板计数;显微镜下观察血小板形态;流式细胞术分析血小板膜蛋白;透...目的分析一先天性血小板减少症家系的临床、实验室特点,并探讨其分子发病机制。方法收集该家系成员的临床资料,采集先证者及其家系成员的静脉血,分别进行全自动及人工血小板计数;显微镜下观察血小板形态;流式细胞术分析血小板膜蛋白;透射电镜观察中性粒细胞胞浆包涵体。聚合酶链反应扩增非肌性肌球蛋白重链9基因(non-muscle myosin heavy chain9gene,MYH9)的40个外显子,分析PCR产物的核苷酸序列,并直接测序确定突变位点。结果镜下观察外周血涂片巨大血小板占90%以上,血小板膜糖蛋白(CD41、CD61、CD42a、CD42b)均在正常范围内,血小板功能正常;中性粒细胞胞浆透射电镜可见无包膜分隔的包涵体,MYH9基因38号外显子第5521位核苷酸存在G→A杂合突变(GAG→AAG),从而导致其编码的非肌性肌球蛋白重链A(NMMHC2A)第1841位谷氨酸变为赖氨酸,正常对照及该家系正常者未见此突变。结论 MYH9基因点突变并伴有血小板减少及巨大血小板是Fechtner综合征的主要特征。展开更多
Objective.To establish a flow cytometric internal standard method for counting platelet-derived microparti-cles(PMPs)and to study its clinical significance. Methods. PMPs suspension(platelet poor plasma,PPP) was extra...Objective.To establish a flow cytometric internal standard method for counting platelet-derived microparti-cles(PMPs)and to study its clinical significance. Methods. PMPs suspension(platelet poor plasma,PPP) was extracted by gradual centrifugation. According to the size of PMPs,3 μm and 0.8μm 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. Results. In 30 healthy donors,the average concentration of resting PMPs was(1.2×105±5.7×104 )/ml and that of activated PMPs was(1.6×106±9.1×105)/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,( 6.1×105±2.5×105 )/ml, ( 6.8×105±3.4×105)/ml and(5.9×105±3.1×105)/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 [1.3×105±6.1×104)/ml,(P >0.05)] .Conclusions. 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.展开更多
文摘目的分析一先天性血小板减少症家系的临床、实验室特点,并探讨其分子发病机制。方法收集该家系成员的临床资料,采集先证者及其家系成员的静脉血,分别进行全自动及人工血小板计数;显微镜下观察血小板形态;流式细胞术分析血小板膜蛋白;透射电镜观察中性粒细胞胞浆包涵体。聚合酶链反应扩增非肌性肌球蛋白重链9基因(non-muscle myosin heavy chain9gene,MYH9)的40个外显子,分析PCR产物的核苷酸序列,并直接测序确定突变位点。结果镜下观察外周血涂片巨大血小板占90%以上,血小板膜糖蛋白(CD41、CD61、CD42a、CD42b)均在正常范围内,血小板功能正常;中性粒细胞胞浆透射电镜可见无包膜分隔的包涵体,MYH9基因38号外显子第5521位核苷酸存在G→A杂合突变(GAG→AAG),从而导致其编码的非肌性肌球蛋白重链A(NMMHC2A)第1841位谷氨酸变为赖氨酸,正常对照及该家系正常者未见此突变。结论 MYH9基因点突变并伴有血小板减少及巨大血小板是Fechtner综合征的主要特征。
文摘Objective.To establish a flow cytometric internal standard method for counting platelet-derived microparti-cles(PMPs)and to study its clinical significance. Methods. PMPs suspension(platelet poor plasma,PPP) was extracted by gradual centrifugation. According to the size of PMPs,3 μm and 0.8μm 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. Results. In 30 healthy donors,the average concentration of resting PMPs was(1.2×105±5.7×104 )/ml and that of activated PMPs was(1.6×106±9.1×105)/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,( 6.1×105±2.5×105 )/ml, ( 6.8×105±3.4×105)/ml and(5.9×105±3.1×105)/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 [1.3×105±6.1×104)/ml,(P >0.05)] .Conclusions. 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.
