一株新的急性髓系白血病细胞系SH-2的建立及其鉴定

目的 采用自体骨髓基质细胞与原代白血病细胞加rhIL-3后共培养的方法建立1株新的人急性髓系白血病(AML)细胞系SH-2,并对其生物学特征进行全面鉴定.方法 采集1例经联合化疗和异基因外周血干细胞移植末缓解的AML-M2a型患者骨髓,分离单个核细胞,加入含20%胎牛血清的IMDM培养基内置37℃、5%CO2、饱和湿度培养箱中传代培养,培养过程中保留自体骨髓基质细胞,同时加入细胞因子rhIL-3,长期体外培养成功建立伴有-Y,der(16)t(16;17)(q24;q12),-17,+19和p53突变的AML细胞系SH-2,并通过细胞学、遗传学、免疫学、分子学和小鼠致瘤实验等多种方法对SH-2细胞的生物学特征进行全面鉴定.结果 SH-2细胞已在体外持续生长3年余而不需加用生长因子和基质细胞.其EB病毒、支原体检测均为阴性.SH-2细胞具有和原代白血病细胞相同的髓系细胞形态学特点,伴有自然杀伤相关抗原表达的AML免疫表型(CD13+、CD33+、CD56+、CD16/56+)和45,X,-Y,der(16)t(16;17)(q24;q12),-17,+19的亚二倍体核型,后者逐渐被伴有上述染色体异常的近四倍体核犁所取代.荧光原位杂交(FISH)和多色FISH证实了以上异常,并揭示-17导敛1个p53基因丢失.DNA序列分析揭示另1个p53等位基因第5号外显子的576编码子点突变(CAC→CAT).RT-PCR显示除了表达于细胞因子(SCF)外,其余细胞因子均不表达;不表达多药耐药基因而表达凋亡相关基因,如bcl-2、Fas、GST-1T和p21.短串联重复序列PCR证明了SH-2细胞和患者门血病细胞的同源性.SH-2细胞有一定的集落形成能力并能在裸鼠皮下及SCID小鼠内脏成瘤.结论 SH-2是一株新的具有明晰生物学背景的AML细胞系,为白血病研究提供了又一有用工具.     

一株新的急性髓系白血病细胞系SH-2的建立及其鉴定

Objective To establish and characterize a novel human myeloid leukemia cell line SH-2. Methods Bone marrow mononuclear cells(BMMNC) isolated from a AML-M2 patient, who failed to ob-tain complete remission after chemotherapy and allogenic bone marrow transplantation were passed in a long term IMDM culture medium supplemented with 20% fetal calf serum. Stromal cells were retained and rh-IL-3was added in the culture system. A new human myeloid leukemia cell line SH-2 was successfully established with a cytogeuetic characteristics of a loss of Y chromosome(- Y), a derivative chromosome 16 resulting from unbalanced translocation between chromosome 16 and 17, monosome 17, trisomy 19 and p53 alteration. Vari-ous methods were employed to characterize SH-2 cell line. Results SH-2 cells has been maintained without cytokine and stromal cells for more than 3 years without EB virus and mycoplasma contamination. SH-2 cells had the basically same morphological, immunophenotypic and cytogenetic features as the patient' s leukemia cells did, such as myeloid morphology, an immunophenotype of CD13+ , CD33+ , CD56+ , CD16/56+ and a hypodiploid karyotype of 45, X, - Y, der(16) t(16;17) (q24;q12) , - 17, + 19, which were gradually de-creased and replaced by the near-tetraploid cells with a karyotype of 73 - 102 (80), XX, - Y, - Y, del (lq31) ×2, der(16)t(16;17) (q24;q12) ×2, - 17, - 17, + 19, + 19. FISH and multiple FISH delineated all the abnormalities and revealed a loss of one p53 allele due to monosomy 17. DNA direct sequencing detec-ted a point mutation of CAG to CAT at codon 576 of exon 5 in another p53 allele. RT-PCR showed that SH-2 cells expressed apoptosis-related genes (bcl-2, Fas, GST- π and p21) rather than MDR-related genes. Short tandem repeat PCR provided powerful evidence for the derivation of SH-2 cell line from the patient' s leukemia cells. SH-2 cells had certain colony formation and tumorigenic capacities in nude and SCID mice. Conclu-sion SH-2 is a new myeloid leukemia cell line with a unique biology background, and will provide a useful tool for leukemia research.     

非血缘关系与单倍型相合供者造血干细胞移植治疗恶性血液病疗效比较

目的 比较非血缘关系供者造血干细胞移植(URD-HSCT)与单倍型相合供者移植(Hi-HSCT)的临床疗效.方法 接受URD-HSCT和Hi-HSCT的血液病患者分别为25例和30例,以改良BUCY方案或TBI/CY方案预处理,以环孢素联合甲氨蝶呤及霉酚酸酯为基础方案预防急性移植物抗宿主病(aGVHD),部分患者加用抗胸腺细胞球蛋白(ATG)或抗淋巴细胞球蛋白(ALG)及CD25单抗.结果 URD-HSCT组均获造血重建,中位随访13个月,3年无病生存率(DFS)(54.1±11.9)%;Hi-HSCT组29例造血重建,中位随访10.3个月,3年DFS(43.1±9.1)%(P=0.13).两组累积Ⅲ～Ⅳ度aGVHD发生率分别为40.0%(10例)和37.9%(11例)(P＞0.05).血液病复发各2例(复发率8.0%和6.0%,P＞0.05),移植相关死亡率分别为40.0%(10例)和56.7%(17例)(P＞0.05),死因依次为重度aGVHD合并感染、重症肺部感染和血液病复发.结论 URD-HSCT及Hi-HSCT均为治疗难治及高危恶性血液病的有效手段,应个体化选择最适供者,重度aGVHD和继发感染仍需要更积极有效的预防措施.     

急性早幼粒细胞白血病的临床和生物学特征,初诊和ATRA治疗中高白细胞患者复发率高

急性早幼粒细胞白血病(APL)为AML的一种特殊亚型,具有特殊的形态特征和染色体易位t(15;17)及PML-RAR α融合基因,根据PML基因断裂点不同分为长型(L),短型(S)。变异型(V)。全反式维甲酸(ATRA)对具有t(15;17)的APL患者的诱导缓解(CR)率高达95％,CR后强烈化疗可使60％的患者长期无病生存,然而仍有40％患者复发,另外ATRA治疗中具有危及生命的维甲酸(RA)综合征。本文旨在阐述APL患者临床及生物学特征(台湾),CD2表达与PML-RAR α     

一株新的急性髓系白血病细胞系SH-2的建立及其鉴定

Objective To establish and characterize a novel human myeloid leukemia cell line SH-2. Methods Bone marrow mononuclear cells(BMMNC) isolated from a AML-M2 patient, who failed to ob-tain complete remission after chemotherapy and allogenic bone marrow transplantation were passed in a long term IMDM culture medium supplemented with 20% fetal calf serum. Stromal cells were retained and rh-IL-3was added in the culture system. A new human myeloid leukemia cell line SH-2 was successfully established with a cytogeuetic characteristics of a loss of Y chromosome(- Y), a derivative chromosome 16 resulting from unbalanced translocation between chromosome 16 and 17, monosome 17, trisomy 19 and p53 alteration. Vari-ous methods were employed to characterize SH-2 cell line. Results SH-2 cells has been maintained without cytokine and stromal cells for more than 3 years without EB virus and mycoplasma contamination. SH-2 cells had the basically same morphological, immunophenotypic and cytogenetic features as the patient' s leukemia cells did, such as myeloid morphology, an immunophenotype of CD13+ , CD33+ , CD56+ , CD16/56+ and a hypodiploid karyotype of 45, X, - Y, der(16) t(16;17) (q24;q12) , - 17, + 19, which were gradually de-creased and replaced by the near-tetraploid cells with a karyotype of 73 - 102 (80), XX, - Y, - Y, del (lq31) ×2, der(16)t(16;17) (q24;q12) ×2, - 17, - 17, + 19, + 19. FISH and multiple FISH delineated all the abnormalities and revealed a loss of one p53 allele due to monosomy 17. DNA direct sequencing detec-ted a point mutation of CAG to CAT at codon 576 of exon 5 in another p53 allele. RT-PCR showed that SH-2 cells expressed apoptosis-related genes (bcl-2, Fas, GST- π and p21) rather than MDR-related genes. Short tandem repeat PCR provided powerful evidence for the derivation of SH-2 cell line from the patient' s leukemia cells. SH-2 cells had certain colony formation and tumorigenic capacities in nude and SCID mice. Conclu-sion SH-2 is a new myeloid leukemia cell line with a unique biology background, and will provide a useful tool for leukemia research.     

异基因造血干细胞移植治疗T淋巴母细胞淋巴瘤/白血病1例并文献复习

目的:探讨T淋巴母细胞淋巴瘤/白血病的临床诊断及治疗方法。方法:报告1例T淋巴母细胞淋巴瘤/白血病的病例,并复习相关文献。结果:患者通过手术病理及骨髓相关检查确诊T淋巴母细胞淋巴瘤/白血病,联合化疗后行母供子半相合造血干细胞移植术,术后1年患者出现复发趋势,予供体淋巴细胞输注后本病再次处于缓解状态,目前患者病情稳定。结论:T淋巴母细胞淋巴瘤/白血病是高度恶性淋巴瘤,一旦进展为白血病阶段,预后恶劣,生存期短,造血干细胞移植联合供体淋巴细胞输注是治疗本病延长生存期的一种有效的方法。     

造血干细胞移植后早期出血并发症的危险因素与预后分析

目的了解造血干细胞移植（HSCT）患者早期出血并发症的状况,分析其危险因素及与生存的关系。方法观察318例HSCT患者的出血状况,按出血程度分为轻、中、重度出血,按出血部位分为皮肤黏膜、消化道、呼吸道、女性阴道、眼、颅内出血及出血性膀胱炎（HC）,选用Logistic回归模型分析各因素与出血的相关性,Cox比例风险模型进行预后分析。结果211例患者在移植后初期发生出血并发症,其中轻度143例（45．O％）,中度59例（18．6％）,重度9例（2．8％）。预处理方案含ATG（OR=3．460）、异基因来源干细胞（OR=1．918）、感染（OR=1．691）、急性移植物抗宿主病（aGVHD）（OR：2．252）、血小板最低值≤15×109／L（OR=2．499）是出血的危险因素,异基因移植（OR=5．431）、aGVHD（OR=5．059）、巨细胞病毒感染（OR：4．241）、多瘤病毒尿（OR=5．723）是HC的危险因素,重度出血（RR=6．106）、出血部位位于颅内（RR=12．131）及呼吸道（RR=9．202）可独立增加移植后死亡风险。结论移植期间血小板减少等多种因素可造成出血风险增加,重度出血、颅内出血及呼吸道出血增加移植后患者死亡风险。     

非T细胞去除单倍体造血干细胞移植治疗T淋巴母细胞性淋巴瘤的初步临床研究

 目的　探讨非T细胞去除单倍体造血干细胞移植（haplo-HSCT）治疗T淋巴母细胞性淋巴瘤（T-LL）的疗效。方法　3例确诊的T-LL患者获得缓解后接受了粒细胞集落刺激因子（G-CSF）动员后的非T细胞去除的单倍体亲缘供体的骨髓干细胞输注。其中2例患者预处理采用包含环磷酰胺（CTX）、全身照射（TBI）和阿糖胞苷（Ara-C）的清髓性方案,另1例患者采用包含CTX和白消安（BU）、Ara-C的清髓性方案;3例患者都采用了包含兔抗人胸腺细胞球蛋白（ATG）的加强的急性移植物抗宿主病预防方案。结果　所有患者均获得快速完全的造血重建,中性粒细胞和血小板中位恢复时间分别为12 d和13 d。3例患者中位随访24个月（9～75个月）,均无瘤生存。结论　非T细胞去除haplo-HSCT治疗T-LL的相关不良反应可以耐受,患者可能获得长期生存。     

伴SET-NUP214融合基因急性白血病异基因造血干细胞移植疗效分析

目的探讨异基因造血干细胞移植(allo-HSCT)治疗伴SET-NUP214融合基因急性白血病(SET-NUP214+AL)的疗效。方法回顾性病例系列研究。收集2014年12月至2021年10月在苏州大学附属第一医院和苏州弘慈血液病医院接受allo-HSCT治疗的18例SET-NUP214+AL患者的临床资料, 分析其疗效及预后, 采用Kaplan-Meier法进行生存分析。结果 18例患者中男性12例, 女性6例, 年龄13～55岁, 中位数29岁。急性混合细胞白血病6例(髓/T表达3例、B/T表达2例、髓/B/T表达1例), 急性淋巴细胞白血病9例(急性B淋巴细胞白血病1例、急性T淋巴细胞白血病8例), 急性髓系白血病3例。所有患者确诊后接受诱导化疗, 17例患者化疗后获得完全缓解, 后续所有患者接受异基因造血干细胞移植。移植前状态:第1次完全缓解15例, 第2次完全缓解1例, 部分缓解1例, 未缓解1例。所有患者完全植入, 中性粒细胞植活和血小板植活的中位时间分别为12、13 d。随访时间4～80个月, 中位数23个月, 18例患者中15例存活, 死亡3例, 死因均为移植后复发。...