CAJ | 학술논문

[目的] 研究非血缘双份脐血移植( DUCBT)的植入规律及植入动力学.[方法] 选择接受DUCBT的血液病患者29例,采用复合扩增荧光标记短串联重复序列结合聚合酶链反应(STR-PCR)方法对供受者16个特异性等位基因进行定量分析,了解造血细胞嵌合体动态变化,判断植入情况,研究DUCBT植入规律.同时,通过对两份脐血复苏后有核细胞总数(TNC)、CD34细胞数、CD3细胞数和NK细胞数、冷冻前集落形成单位(CFU)和粒-巨噬细胞集落形成单位(CFU-GM)进行分析,研究植入份脐血的植入动力学和植入份脐血的特点.[结果] 29例患者中,23例获得植入,其中22例为单份脐血的优势植入,1例为两份脐血的均势植入.22例单份脐血优势植入者中,20例移植后14d、另2例移植后21dSTR-PCR检测呈100％单份脐血嵌合体；6例未获得植入的患者,移植后14 d STR-PCR显示2例呈双份脐血嵌合体,4例呈双份脐血或单份脐血与受者混合嵌合体,移植后21 d外周血和移植后30 d骨髓均呈100％受者嵌合体.DUCBT后7 d STR-PCR检测结果与是否植入不一致(kappa=0.112,P=0.198),14、21d外周血和30d骨髓标本均一致(kappa=0.811,P=0.001；kappa=0.900,P=0.001；kappa=0.900,P=0.001).DUCBT获得植入患者植入份与非植入份脐血相比,复苏后TNC、复苏后CD34细胞数、冷冻前CFU、冷冻前CFU-GM、复苏后CD;细胞数、复苏后NK细胞数差异均无统计学意义(P=0.783、0.455、0.615、0.534、0.114、0.463).[结论] 采用STR-PCR技术检测供受者16个特异性等位基因的定量分析方法确定DUCBT后的植入状态是敏感和特异的,DUCBT后14d可作为确定脐血是否植入的时间.DUCBT中优势植入份脐血的特点仍不清楚,需进一步研究.
[목적] 연구비혈연쌍빈제혈이식( DUCBT)적식입규률급식입동역학.[방법] 선택접수DUCBT적혈액병환자29례,채용복합확증형광표기단천련중복서렬결합취합매련반응(STR-PCR)방법대공수자16개특이성등위기인진행정량분석,료해조혈세포감합체동태변화,판단식입정황,연구DUCBT식입규률.동시,통과대량빈제혈복소후유핵세포총수(TNC)、CD34세포수、CD3세포수화NK세포수、냉동전집락형성단위(CFU)화립-거서세포집락형성단위(CFU-GM)진행분석,연구식입빈제혈적식입동역학화식입빈제혈적특점.[결과] 29례환자중,23례획득식입,기중22례위단빈제혈적우세식입,1례위량빈제혈적균세식입.22례단빈제혈우세식입자중,20례이식후14d、령2례이식후21dSTR-PCR검측정100％단빈제혈감합체；6례미획득식입적환자,이식후14 d STR-PCR현시2례정쌍빈제혈감합체,4례정쌍빈제혈혹단빈제혈여수자혼합감합체,이식후21 d외주혈화이식후30 d골수균정100％수자감합체.DUCBT후7 d STR-PCR검측결과여시부식입불일치(kappa=0.112,P=0.198),14、21d외주혈화30d골수표본균일치(kappa=0.811,P=0.001；kappa=0.900,P=0.001；kappa=0.900,P=0.001).DUCBT획득식입환자식입빈여비식입빈제혈상비,복소후TNC、복소후CD34세포수、냉동전CFU、냉동전CFU-GM、복소후CD;세포수、복소후NK세포수차이균무통계학의의(P=0.783、0.455、0.615、0.534、0.114、0.463).[결론] 채용STR-PCR기술검측공수자16개특이성등위기인적정량분석방법학정DUCBT후적식입상태시민감화특이적,DUCBT후14d가작위학정제혈시부식입적시간.DUCBT중우세식입빈제혈적특점잉불청초,수진일보연구.
[Objective]To study the discipline of implantation and implantation dynamics in unrelated double umbilical cord blood transplantation (DUCBT).[Methods]Twenty-nine patients with hematologic malignancies who undergoing two-units unrelated donor cord blood transplantation were included in the study.After transplantation,hematopoietic chimerism of peripheral blood was evaluated by the Results of short tandem repeat with polymerase chain reactions (STR-PCR) which quantitatively determinated 16 specific alleles between donor and receptor, to find out their chimerism dynamic change, to judge whether transplantation was implanted and judge which one was implanted,and to study the discipline of implantation in DUCBT.At the same time,total nucleated cells(TNC),dose of CD34 cells,colony forming unit(CFU),colony forming unit-granulocyte and macrophage(CFU-GM),dose of CD; cells,dose of natural killer(NK)cells were compared between dominant units and non-dominant ones,to quest the discipline implantation dynamics of DUCBT.[Results]In 29 clinical cases,23 cases obtained engraftment,including 22 cases appearing one unit cord blood engraftment and 1 case appearing two units cord blood engraftment.Of 22cases with one dominant unit engraftment,at 14 days after DUCBT,the results of STR-PCR showed that 20cases appeared one dominant unit engraftment,other 2 cases appeared one dominant unit engraftment at 21days after DUCBT.Of 6 cases without engraftment,at 14 days after DUCBT,2 cases showed chimerism of two units cord blood,other 4 cases showed chimerism of two units cord blood or one unit cord mixed with receptor.At 30 days after DUCBT,their STR-PCR results of bone marrow showed full donor chimerism.Compared results at day 7,day 14,day 21 by peripheral blood,and day 30 by bone marrow with results of implantation after DUCBT,their coherence were kappa=0.112,P=0.198,kappa =0.811,P =0.001,kappa =0.900,P =0.001 and kappa =0.900,P =0.001,respectively.In addition,compared dominant unit with nondominant unit,TNC,doses of CD+34 cells,CFU,CFU-GM,CD; cells and NK cells were all no significant difference between them (P=0.783,0.455,0.615,0.534,0.114,0.463,respectively).[Conclusion]STR-PCR which quantitatively determinates 16 specific alleles between donor and receptor is sensitively and specifically to judge implant status.The 14 days after DUCBT was the time when implant is embedded.However,the implantation dynamics of DUCBT is still unknown which need further quest in the future.