骨髓增生异常综合征相关性急性淋巴细胞性白血病(附7例报告)

目的 :探讨骨髓增生异常综合征相关性急性淋巴细胞白血病 ( MDS/ AL L)的特点。方法 :MDS/AL L与骨髓增生异常综合征相关性急性髓细胞性白血病 ( MDS/ AML )的临床及实验室特点进行比较 ,采用 t检验、检验统计。结果 :MDS转化成 AL L 8.14 % ,其类型为 型 5例 , 型 2例。 AL L类型 L1 1例 ,L2 6例。临床及血象在 MDS/ AML与 MDS/ AL L二组之间无明显差异。骨髓象二组之间差异较大。 MDS/ AL L在 MDS期部分类似一过性再生障碍性贫血 ,骨髓幼稚细胞均值为 0 .74 6,高于 MDS/ AML,( P<0 .0 1)。红系为 0 .0 8,低于 MDS/AML( P<0 .0 1)。巨核细胞低于 MDS/ AL L,但无明显差异。MDS/ AL L 在 MDS期及 AL L 期均罕见形态学改变。结论 :MDS/ AL L与 MDS/ AML有异质性。这与干细胞受累的水平不同有关 ,MDS/ AL L可能是在多能造血干细胞时受累     

表皮生长因子受体与骨髓异常造血关系的研究

目的　阐明表皮生长因子受体 (EGFR)与人类骨髓造血细胞异常增生和异常发育的关系。方法　对5 6例恶性血液病、2 9例良性血液病及 9例正常人 ,应用 Southern Blot技术检测骨髓细胞中 EGFR基因的扩增与重排 ,应用免疫酶标技术检测骨髓细胞 EGFR的表达情况。结果　所检测的标本均未发现 EGFR基因的扩增及表达 ,仅在 2例急性白血病 (1例 M3 、1例 M6)中发现可疑重排带 ,2例重排带的大小相同均为 3 .3 kb。结论　 EGFR基因的扩增与重排较少发生在人类血液系统恶性疾患中。人类良性和恶性骨髓造血细胞均无 EGFR的表达。提示EGFR表达异常可能较少与人类造血细胞的增殖与发育异常有关。在骨髓增殖异常综合征 (MDS)和白血病中可能存在发生扩增与异常表达的非 EGFR基因的其它与 v- erb B同源的基因 ,参与 MDS和白血病的发生与发展。     

内源性逆病毒癌基因V-erbB突变与白血病发生

目的：证明鸡原始红细胞增多症逆病毒癌基因V—erbB在人类白血病和骨髓增生异常综合征(MDS)急者基因组内的存在。方法：应用V—erbB PCR，V—erbB寡核苷酸(Oligo)原位杂交技术和PCR产物DNA测序，对84例MDS、可疑MDS和49例其他相关血液病进行检到。结果：2例患者骨殖细胞基因组内均存在V—erbB亚基因片段，同源性占99．5％，它们具有相同的限制性酶勿位点，因此可以在Southern印迹杂交条件下发生骨髓细胞C—erbB重排和重排／扩增。白前P1P2PCR产物有420bp，P1P3产物有650bp，而白血病(AL—M4)P1P2PCR产物仅390bp，P1P3PCR因P3无模板序列而无结果。提示从白前发展为白血病，可能发生V—erbB亚基因序列的缺失，即缺失突变。结论：大鼠和人白血病和食管癌等可能均起源于其基因组内存在V—erbB亚基因及其缺失突变。白血病发病中除了上述病因以外，骨髓细胞染色体不稳定性可能起协同作用。     

骨髓增生异常综合征骨髓细胞端粒酶活性和端粒酶逆转录酶的表达

目的　探讨骨髓增生异常综合征 ( MDS)骨髓细胞端粒酶活性与端粒酶逆转录酶表达的关系及临床意义。方法　采用端粒重复序列扩增 -酶联免疫吸附试验 ( TRAP- EL ISA)检测端粒酶活性 ,RT- PCR法检测端粒酶逆转录酶 ( h TERT) m RNA水平的表达。结果　高危组 MDS患者骨髓细胞端粒酶活性较低危组 MDS患者骨髓细胞端粒酶活性明显增高 ( 0 .2 0 0± 0 .2 2 5 vs 0 .0 2 7± 0 .0 13,P=0 .0 37) ;正常人、缺铁性贫血 ( IDA)患者和低危组 MDS患者骨髓细胞 h TERT m RNA表达检测结果均阴性 ,而高危组 MDS患者和急性白血病 ( AL )患者骨髓细胞均表达h TERT基因 ;h TERT m RNA表达和端粒酶活性呈正相关 ( r=0 .774 ,P=0 .0 2 4 )。结论　端粒酶活性与 h TERT基因 m RNA水平的异常表达能反映出 MDS病情的进展。定期检测 MDS患者骨髓细胞端粒酶活性和 h TERTm RNA水平的表达可作为观察 MDS患者病情变化的指标之一。     

C—erbB2、C—myc及EGFR基因共扩增与非小细胞肺癌预后的研究

目的：探讨C-erbB2、C-myc和EGFR多种癌基因共扩增与人体非小细胞肺癌的复发、转移和预后的相关性.方法：采用DNA印迹技术（Blot）分别检测156例非小细胞肺癌手术标本及正常肺组织DNA中C-erbB2、C-myc和EGFR基团扩增现象，并对上述所检测的患者作5年随访追踪以观察多种癌基因扩增与肺癌患者预后的相关性。结果：C-erbB2、C-myc和EGFR基因中出现双基因以上的共扩增率与肺癌的TNM分期呈正相关，I期患者的共扩增率为46％，Ⅱ～Ⅲ期肺癌患者的阳性共扩增率为64％（P＞0．05）。多种癌基因共扩增呈阴性的肺癌患者生存率高于呈阳性患者，两组间具有明显差异（P＝0．001）。多种癌基因共扩增率与肺癌的组织学类型、性别和年龄无相关性（P＞0．05）。结论：C-erbB2、C-myc和EGFR多基因共扩增率与非小细胞肺癌患者的复发、转移和生存期相关。     

CAT方案治疗难治性急性髓系白血病的临床观察

目的：初步观察CAT(环磷酰胺、阿糖胞苷、拓扑替康)方案对难治性急性髓系白血病(AML)的近期临床疗效并评价此方案的不良副作用。方法：选择8例难治性AML(原发难治性AML3例，AML伴多系形态发育异常，此前有MDS病史患者3例，慢性粒细胞白血病AML变2例)．应用CAT方案治疗，其中4例治疗1疗程，其余4例治疗2疗程。结果：1例在骨髓抑制期死于感染性休克．可评价疗效7例，1例达完全缓解(CR)，3例达部分缓解(PR)，1例CML急变患者经2疗程后回到慢性期，总有效率71．4％(5/7例)，中位生存期为6．5(0．3—22^ )个月。主要不良副作用为骨髓抑制。结论：CAT方案为高危MDS、加速／急变期CML和继发于MDS的AML患者的一个有效且毒性可耐受的治疗新方案。     

c-FLIP mRNA 在恶性血液病中的表达及其意义

目的：探讨C—FLIPmRNA在恶性血液病中的表达及其意义。方法：用采用半定量逆转录聚合酶链反应（RT—PCR）检测42例恶性血液病骨髓单个核细胞C—FLIPmRNA的表达。包括急性白血病（AL）27例，其中初治21例及复发和完全缓解（CR）后AL各3例、慢性粒细胞性白血病（CML）5例、慢性中性粒细胞性白血病（CNL）1例和慢性淋巴细胞性白血病（CLL）4例，多发性骨髓瘤（MM）3例，骨髓增生异常综合征-难治性贫血伴原始细胞增多2型（MDS—RAEB-2）2例。结果：在初治和复发AL、初治CML、CNL、CLL、MDS—RAEB-2、MM中C—FLIPmRNA均呈异常增高表达，初治AL中c—FLIPmRNA的表达与复发AL比较差异无统计学意义（P〉0．05），其FAB各亚型之间的表达差异亦无统计学意义（P〉0．05）。初治AL与CLLC—FLIPmRNA的表达显著高于初治CML（P〈0．001），但初治AL与初治CLLE魄磋淠呒统计学意义（P〉0．05）。MDS—RAEB-2、MMC—FLIPmRNA的表达与AL的cFLIPmRNA表达均无统计学差异（P〉0．05）。对照组和CR后AL均为阴性表达。C—FLIPmRNA的表达与初治AL患者年龄、性别、初诊白细胞数、LDH以及核型、免疫表型无关。初治未达CR的AL患者其c—FLIPmRNA表达高于CR者，但并无统计学意义（尸〉0．05）。结论：恶性血液病C—FLIPmRNA的表达异常增高。C—FLIPmRNA能反映恶性血液病骨髓细胞的凋亡抑制情况，并与恶性血液病的类型、疾病状态、临床疗效和预后密切相关。     

bcl-2和bax基因在急性白血病细胞中的表达及其意义

目的：研究bcl-2和bax基因在急性白血病(AL)细胞中的表达及其与AL的分型、临床特征、疗效及预后因素的关系。方法：采用免疫组化SP法检测54例初治AL骨髓细胞的bcl-2和bax基因蛋白的表达情况，分析其与FAB分型、临床特征、疗效及预后因素的关系。结果：AL骨髓细胞中bcl-2基因的表达显著高于正常对照组(P＜0.05)，而bax基因的表达低于正常对照(P＜0.05)。bcl-2和bax基因表达在急性髓系白血病(AML)与急性淋巴细胞白血病(ALL)无显著差异；在AML的亚型中，bcl-2在M．及M5中的表达高于M1、M2和M3(P＜0.05)；而bax的表达在各亚型中无显著性差异。bcl-2的表达与初诊时的白细胞数呈正相关(r=0.44，P＜0.05)，与疗效呈负相关(r=-0.40，P＜0.05)；bax的表达与骨髓白血病细胞数呈负相关(r＝-0.34，P＜0.05)，与髓外浸润呈负相关(r＝-0.45，P＜0.05)。结论．bcl-2和bax基因的紊乱是AL形成与发展的原因之一。这些细胞凋亡调控基因不仅参与了白血病的形成，还与白血病的某些临床特征、疗效及预后因素密切相关。     

AML／TMDS与MDS—AML的对比研究

目的：分析三系病态造血的急性髓系白血病(AML／TMDS)与骨髓增生异常综合征演变为急性髓系白血病(MDS—AML)的差异。方法：采用常规形态学和细胞化学染色方法观察患者外周血和骨髓细胞。结果：AML/TMDS患者血小板计数和外周原始细胞百分率高于MDS—AML(P＜0．05，P＜0．01)；小巨核细胞和Pseudo-pelger异常明显低于MDS-AML(P＜0．05，P＜0．01)，2组病例MPO染色阳性率基本一致，但AML/TMDS患者MPO积分显著高于MDS—AML(P＜0．01)。AML/TMDS患者多核原始红细胞明显低于MDS—AML(P＜0．05)；AML／TMDS与MDS—AML患者巨大血小板分别为23．1％和86．7％。结论：AML/TMDS与MDS—AML白血病克隆不同，正确诊断对确定方案、判断疗效及预后有重要意义。     

骨髓增生异常综合征和再生障碍性贫血等骨髓TNF-α活性与白血病的关系

目的 :探讨免疫抑制因素 TNF- α在白血病发病中的作用。方法 :在大系列 MDS细胞遗传学研究基础上 ,应用酶联免疫吸附法 (EL ISA)对 2 3例 MDS、17例 AA、10例 AL 和 8例 ITP等患者骨髓液进行 TNF- α活性水平的测定。结果 :MDS TNF- α活性水平有明显增加、AL 则有显著性增加 (P<0 .0 5 )并同它们中染色体核型异常和 SCD阴性检出率的增加相一致。AA与正常人差别不明显 ,ITP与正常人几乎无差别 ,这同它们中大多数患者骨髓细胞核型正常和 SCD阳性的结果相一致。部分病例随访结果显示 ,当 MDS转白血病时不仅其骨髓细胞 SCD由阳性转阴性 ,其 TNF- α活性水平呈数倍于正常值的扩增。结论 :尽管 MDS和 AA同属造血干细胞病并有共同的遗传背景和发病早期 ,但前者主要还与体液免疫抑制因素 (TNF- α)相关 ,而后者则还与细胞免疫抑制因素 (如 T-淋巴细胞 )相关 ,因而有不同的表现。     

The prognostic significance of bone marrow levels of neurofibromatosis-1 protein and ras oncogene mutations in patients with acute myeloid leukemia and myelodysplastic syndrome

BACKGROUND: It has been reported that point mutations of the ras gene occur frequently in acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS). However, the prognostic significance of ras gene mutations in patients with these disorders has been a controversial issue. Although abnormalities in the neurofibromatosis 1 (NF1) gene, which is a gene involved in the ras pathway, have been observed frequently in patients with juvenile chronic myelogenous leukemia, the role of these abnormalities in adult patients with AML or MDS is not clear. METHODS: In this study, bone marrow specimens that were obtained from previously untreated patients with AML and MDS were examined for ras mutations, and the levels of NF1 protein expression were measured. RESULTS: Ras point mutations were detected in 16 of 83 patients with AML (19%) and in 21 of 90 patients with MDS (23%). Fourteen of 28 patients with chronic myelomonocytic leukemia (50%) had ras gene mutations. Decreased bone marrow levels of NF1 protein (< 70% of the median level detected in control bone marrow specimens) were observed in 20 of 66 patients with AML (30.3%) and in 8 of 29 patients with MDS (27.6%); none of the patients with ras gene mutations had decreased bone marrow levels of NF1 protein. The presence of a mutant ras gene was associated with a shorter complete remission duration (CRD) in patients with MDS (P = 0.05) but had no effect on survival in patients with AML or MDS. Patients with AML who had decreased NF1 protein levels had a slightly longer CRD compared with patients who had normal NF1 levels (P = 0.07). However, the NF1 level had no significant impact on survival among patients with AML or MDS. When patients with low NF1 levels were grouped with patients who had ras mutations, the patients who had AML had a significantly longer CRD compared with the patients who had MDS (P = 0.02). CONCLUSIONS: The current results suggest that a reduction in NF1 protein expression and the presence of ras point mutations may complement each other and that they both play a complex role in the biology of AML and MDS.     

骨髓中CDKN1C阳性表达在MDS和AML患者中检测的临床意义

目的 探讨骨髓中CDKN1C阳性表达在骨髓增生异常综合征(MDS)和继发性急性髓系白血病(AML)患者中检测的临床意义。方法 选取125例MDS/AML患者作为研究对象,同时选取20例健康人群作为健康对照组,分析MDS/AML患者骨髓CD34⁺细胞中CDKN1C的mRNA和蛋白表达水平,比较不同CDKN1C表达水平的MDS患者生存率,采用Cox回归分析MDS和AML患者生存率影响因素,并分析治疗方法对不同CDKN1C表达水平的MDS患者生存率的影响。结果 MDS/AML患者骨髓CD34⁺细胞中CDKN1C mRNA和蛋白表达水平显著高于健康对照组,且与BM(Bone marrow)比例呈正相关(r=0.423,P=0.012);CDKN1C高表达组患者的生存率显著低于CDKN1C低表达组和CDKN1C中表达组(P＜0.05);Cox回归分析结果显示高龄、高BM比例、细胞遗传学风险差以及CDKN1C阳性显著影响MDS/AML患者生存率(P＜0.05);MDS/AML化疗的CDKN1C阳性表达组患者生存率显著低于CDKN1C阴性组患者(P＜0.05),MDS/AML化疗后造血干细胞移植(AlloSCT)的CDKN1C阳性组患者生存率显著低于CDKN1C阴性组患者(P＜0.05)。结论 MDS/AML患者骨髓中CDKN1C的表达显著增高,CDKN1C阳性表达显著影响化疗MDS/AML患者的预后生存。     

再生障碍性贫血和克隆演变:种系和体细胞遗传学

对于骨髓衰竭(BMF)的患者,克隆演变为骨髓增生异常综合征(MDS)或急性髓系白血病(AML)仍然是一个严重的并发症.明确演变为MDS或者AML的危险因素可以为个体化治疗方案提供信息,并确定早期或前期进行造血干细胞移植可能受益的患者.目前,临床实验室可以进行二代DNA测序,研究集中在种系和体细胞突变对BMF患者诊断和监测的应用.大多数种系遗传性BMF疾病具有高倾向MDS或AML转化的特征.近来BMF的体细胞突变研究发现高频的克隆性造血干细胞具有获得性基因突变,这些基因与MDS或AML相关.文章就第58届美国血液学会(ASH)年会中对于BMF患者进展为克隆性疾病的种系和体细胞突变的评价以及临床基因检测的挑战进行介绍.     

T-cell receptor beta chain gene rearrangement in acute myeloid leukemia always occurs at the allele that contains the undermethylated J beta 1 region.

The epigenetic phenomenon could play a role in the interaction between chromatin and DNA-binding enzymes, allowing us to consider an association between the phenomenon and gene rearrangement. The correlation between methylation status and rearrangement of the T-cell receptor (TCR) beta chain gene in leukemia cells obtained from patients with acute myeloid leukemia (AML) was examined. All of the AML patients with a TCR-beta rearrangement had hypomethylated CCGG sequences within the J beta 1 region on the rearranged allele, while the germline allele had completely methylated CCmeGG sequence in this region, indicating a strong association between hypomethylation status and rearrangement of the TCR beta chain gene. In the DNA from AML patients with or without a TCR-beta rearrangement, the C beta 2 region contained completely methylated CCmeGG sequences, even though they express T-cell-associated antigens, including CD7; this pattern is quite different from that observed in T-cell neoplasias. Moreover, some AML patients showed a TCR-beta rearrangement without the presence of immunoglobulin heavy-chain gene rearrangement, suggesting that TCR beta chain gene involvement in AML is required for unknown factors other than common recombinase activity.     

荧光原位杂交技术检测c-erbB-2和p53基因在胃癌中的表达

目的：探讨胃癌组织中ｃ－ｅｒｂＢ－２和Ｐ５３基因的表达及其意义。方法：应用荧光原位杂交技术对５５例有癌及移行区、正常区组织的常规石蜡包埋标本进行检测。结果：ｃ－ｅｒｂＢ－２和Ｐ５３基石 阳性率分别为３６．３６％和４５．４５％,移行芡为１０．９１％和３．６４％,正常粘膜区为３．６５％和０,三区间差异无有极显著意义（Ｐ值均〈０．０１）。在肠型和弥温型胃癌中,ｃ－ｅｒｂＢ阳性率分别为５１．６１％和１６．     

Prevalence of N-ras mutations in children with myelodysplastic syndromes and acute myeloid leukemia.

The ras proto-oncogene family encodes a group of 21 kDa nucleotide-binding proteins. Activating mutations of ras genes are associated with certain types of malignancies, indicating that they are related in some way to the malignant process. We have examined bone marrow cells from nine children with myelodysplastic syndromes (MDS) and 35 with acute myeloid leukemia (AML) for activating point mutations of ras genes by in vitro amplification using polymerase chain reaction (PCR), oligonucleotide hybridization and sequencing of PCR products. We found N-ras mutations in cells from 3 of 9 children (33%) with MDS and only 2 of 35 children with AML (6%; 95% confidence interval is 0.7-19%). All mutations the second nucleotide of codon 12 or the first nucleotide of codon 61 of N-ras. There was no apparent correlation with clinical or laboratory characteristics, including karyotype; however, an association of N-ras activation with the most aggressive type of MDS was noted. Among the patients with MDS, 2 of 6 with monosomy 7 had N-ras mutations; however, three children with monosomy 7 which presented with AML lacked ras mutations. One patient was studied at time of diagnosis of MDS and again after progression to AML. At the preleukemic stage of disease, an N-ras mutation was identified; however, after development of AML this mutation was not present in the leukemic clone. In conclusion, these data show that ras mutations, while not necessary for leukemic transformation, may be important for the initiation of preleukemias evolving into overt AML.     

More cell death in refractory anemia with excess blasts in transformation than in acute myeloid leukemia.

Refractory anemia with excess blasts in transformation (RAEB-T) is a subgroup of myelodysplastic syndrome (MDS) in which the bone marrow blast count ranges from 20% to 30%. The recently proposed World Health Organization Classification of Hematologic Malignancies eliminated this category from MDS by lowering the blast count cutoff for acute myeloid leukemia (AML) from 30% to 20%. However, MDS is distinguished from AML by a significant increase in apoptosis. To investigate the difference in apoptosis between RAEB-T, AML, and other categories of MDS, we prospectively analyzed fresh bone marrow samples using the Annexin V and mitochondrial potential assays. There was a significantly higher level of apoptosis in RAEB-T than in AML according to both assays, while no significant differences between RAEB-T and other categories of MDS were noted. The data suggest that RAEB-T is more likely to be an advanced stage of MDS and biologically different from AML.     

PATTERN OF T-CELL RECEPTOR DELTA GENE REARRANGEMENT BY SOUTHERN BLOTTING AND POLYMERASE CHAIN REACTION TECHNIQUE IN HONG KONG CHINESE PATIENTS WITH NON-T-CELL HEMATOLOGICAL MALIGNANCIES

It has been recognized that clonal T-cell receptor delta (TCRδ) gene rearrangement is present in both T- and B-cell malignancies. The highly sensitive polymerase chain reaction (PCR) technique may be applicable to cases of leukemia and lymphoma of non-T-cell origin for detection of minimal residual disease (MRD). A PCR technique was used in this study to investigate the pattern of clonal TCRδ gene rearrangement in Hong Kong Chinese patients with non-T-cell hematological malignancies. Seventy-three patients with the diagnosis of acute leukemia and non-Hodgkin's lymphoma of non-T-cell origin were included in this study. There were 20 patients with common ALL (cALL), seven precursal B-cell ALL (PreB-ALL), 23 acute myeloid leukemia (AML), and 23 non-Hodgkin's lymphoma of B-lineage (B-NHL). Clonal rearrangement was detectable by Southern analysis using a Jδ1 probe in 41 per cent of ALL of B-lineage but in none of the B-NHL or AML. The samples were also studied further by monoclonal PCR amplification for TCRδ gene rearrangement. Three different sets of primers were employed to detect clonal rearrangement of the TCRδ gene. The Vδ1(D)Jδ1 recombination typically seen in T-cell malignancies were not seen in any of the of the non-T-cell malignancies. The Vδ2(D)Dδ3 recombination was found exclusively in ALL of B-lineage and was seen in 73 per cent of the Southern positive cases. Although clonal TCRδ gene rearrangement was undetectable by Southern analysis in our AML cases, 26 per cent had a Vδ2(D)Jδ1 recombination found by the PCR technique. Sensitivity of the PCR technique was determined by serial mixing and was up to 5–10 leukemic cells per 10⁴ nucleated cells. It was apparent from this study that it was feasible to detect clonal TCRδ gene rearrangement by the PCR technique in a proportion of the cases of non-T-cell hematological malignancies. The PCR technique can be applied to detect residual leukemic cells in marrow of patients in an apparent morphological complete remission. The value of this application requires further clinical evaluation and correlation.