Abstract:Objective To investigate whether artificial heterozygous chimeras of platelets can be established by intrauterine transplantation of fetal liver stem cells and evaluate its potential use for the treatment of ...Abstract:Objective To investigate whether artificial heterozygous chimeras of platelets can be established by intrauterine transplantation of fetal liver stem cells and evaluate its potential use for the treatment of Glanzmann thrombasthenia.Methods Platelet glycoprotein (GP) Ⅱb Bak a/b (or GPⅡb Ⅰle843Ser) was used as a genetic marker. A homozygous 16-week-old Bak a/a fetus (as donor) and a homozygous 16.5-week-old Bak b/b fetus (as recipient) were screened from 42 pregnant women hospitalized for abortion. PCR with allele specific primers and FOK Ⅰ digestion based on PCR products were used. Aborted donor fetal liver cell suspensions were prepared and intrauterine transplantation was carried out by infusion of 4?ml fetal liver cells (22×105) into the recipient umbilical vein under ultrasonic visualization.Results At gestation termination (abortion), 21 days after transplantation, chimera GPⅡb Bak a/b of the recipient were detected by FOK 1 digestion based on PCR from DNA and RT-PCR from platelet RNA. Conclusion Intrauterine transplantation of fetal liver cell may provide an effective way for curing GT or other inherited diseases.展开更多
目的研究重组人白细胞介素13(rhIL-13)诱导巨核细胞白血病细胞株Dami细胞STAT6(signal transducers and activators of transcription 6)磷酸化及对Dami细胞GPⅡb表达的影响。方法RT-PCR检测Dami细胞IL-13Rα1 mRNA、IL-4RαmRNA和GPⅡb...目的研究重组人白细胞介素13(rhIL-13)诱导巨核细胞白血病细胞株Dami细胞STAT6(signal transducers and activators of transcription 6)磷酸化及对Dami细胞GPⅡb表达的影响。方法RT-PCR检测Dami细胞IL-13Rα1 mRNA、IL-4RαmRNA和GPⅡb mRNA;Western blot检测Dami细胞磷酸化STAT6蛋白表达;流式细胞仪检测Dami细胞GPⅡb蛋白表达。结果Dami细胞表达IL-13 Rα1 mRNA;Dami细胞表达IL-4RαmRNA;IL-13作用Dami细胞2 h,STAT6磷酸化;IL-13组Dami细胞GPⅡb表达高于空白对照组(P<0.05)。结论IL-13诱导Dami细胞STAT6磷酸化并上调Dami细胞GPⅡb表达。展开更多
目的探讨GPⅡb/Ⅲa拮抗剂RGDS和钙通道拮抗剂Nifedipine对ADP诱导的血小板GPⅡb/Ⅲa活化的影响。方法 GP II b/11I a特异性抗体PAC-1标记活化GP II b/11I a,流式细胞仪检测静息及ADP(5μmol/L)诱导的血小板PAC-1表达率。结果静息状态下...目的探讨GPⅡb/Ⅲa拮抗剂RGDS和钙通道拮抗剂Nifedipine对ADP诱导的血小板GPⅡb/Ⅲa活化的影响。方法 GP II b/11I a特异性抗体PAC-1标记活化GP II b/11I a,流式细胞仪检测静息及ADP(5μmol/L)诱导的血小板PAC-1表达率。结果静息状态下的血小板PAC-1表达率为(0.80±0.85)%;在ADP(5μmol/L)激动下,PAC-1表达率为(77.27±9.47)%;RGDS以浓度依赖性的方式抑制PAC-1表达率,IC50值为206μmol/L;Nifedipine能以浓度依赖性的方式抑制PAC-1表达率,IC50值为178μmol/L;RGDS联合Nifedipine对PAC-1表达的联合抑制率为(60.15±16.35)%,二者的交互效应差异有统计学意义(P<0.05)。结论 GPIIb/IIIa受体拮抗剂RGDS和Ca2+拮抗剂Nifedipine均抑制ADP诱导的血小板GPIIb/IIIa活化,其抑制作用呈浓度依赖性,随拮抗剂浓度增加抑制作用增强;两者对血小板GPIIb/IIIa活化的抑制存在协同作用。展开更多
基金ThisworkwassupportedbyNaturalScienceFoundationofHunan Province (No 1995 2 5 2 3)
文摘Abstract:Objective To investigate whether artificial heterozygous chimeras of platelets can be established by intrauterine transplantation of fetal liver stem cells and evaluate its potential use for the treatment of Glanzmann thrombasthenia.Methods Platelet glycoprotein (GP) Ⅱb Bak a/b (or GPⅡb Ⅰle843Ser) was used as a genetic marker. A homozygous 16-week-old Bak a/a fetus (as donor) and a homozygous 16.5-week-old Bak b/b fetus (as recipient) were screened from 42 pregnant women hospitalized for abortion. PCR with allele specific primers and FOK Ⅰ digestion based on PCR products were used. Aborted donor fetal liver cell suspensions were prepared and intrauterine transplantation was carried out by infusion of 4?ml fetal liver cells (22×105) into the recipient umbilical vein under ultrasonic visualization.Results At gestation termination (abortion), 21 days after transplantation, chimera GPⅡb Bak a/b of the recipient were detected by FOK 1 digestion based on PCR from DNA and RT-PCR from platelet RNA. Conclusion Intrauterine transplantation of fetal liver cell may provide an effective way for curing GT or other inherited diseases.
文摘目的探讨GPⅡb/Ⅲa拮抗剂RGDS和钙通道拮抗剂Nifedipine对ADP诱导的血小板GPⅡb/Ⅲa活化的影响。方法 GP II b/11I a特异性抗体PAC-1标记活化GP II b/11I a,流式细胞仪检测静息及ADP(5μmol/L)诱导的血小板PAC-1表达率。结果静息状态下的血小板PAC-1表达率为(0.80±0.85)%;在ADP(5μmol/L)激动下,PAC-1表达率为(77.27±9.47)%;RGDS以浓度依赖性的方式抑制PAC-1表达率,IC50值为206μmol/L;Nifedipine能以浓度依赖性的方式抑制PAC-1表达率,IC50值为178μmol/L;RGDS联合Nifedipine对PAC-1表达的联合抑制率为(60.15±16.35)%,二者的交互效应差异有统计学意义(P<0.05)。结论 GPIIb/IIIa受体拮抗剂RGDS和Ca2+拮抗剂Nifedipine均抑制ADP诱导的血小板GPIIb/IIIa活化,其抑制作用呈浓度依赖性,随拮抗剂浓度增加抑制作用增强;两者对血小板GPIIb/IIIa活化的抑制存在协同作用。
