恶性髓系血液病-7/7q-异常的分子细胞遗传学分析

目的 分析染色体-7／7q-在骨髓增生异常综合征(myelodysplastic syndrome，MDS)和急性髓细胞白血病(acute myeloblastic leukemia，AMI，)中的发生频率；探讨荧光原位杂交技术(fluorescence in situ hybridization，FISH)在检测和鉴定-7／7q-异常中的价值。方法 回顾性分析所有接受细胞遗传学分析(conventional cytogenetic analysis，CCA)的MDS／AML患者的核型特征，其中70份进行FISH分析。应用双色荧光直接标记的7号着丝粒探针(CEP7，光谱绿)和7q31基因序列探针(D7S486，光谱桔红)，15份正常样本作为对照。结果 -7／7q-在AML和MDS中出现频率分别为4．5l％(31／687例)和5．7l％(28／490例)，分别占异常核型病例的5．68％和l0．29％。7q-常见的缺失区域为7q21—22(10例)和7q31—35(10例)。FISH证实伴有克隆性-7／7q-异常，但在随机性-7／7q-异常或正常核型中未检出-7／7q-异常。在核型分析出现7q-异常的病例中，FISH检出7／11例可同时伴有-7克隆的出现，而且7q-异常的细胞数显著高于-7异常细胞数(42．5％vs8．4％，P=0．025)。1例核型为del(7)(q22)患者FISH证实为染色体易位；1例7q 患者FISH显示dup(7q)；1例复杂异常核型，FISH确定其累及7q。结论 FISH是鉴定或确定7q结构异常的强有力工具，能精确地评价-7／7q-。7q-异常通常与-7异常在同一个样本中共存，且7q-细胞数显著增高，推测-7克隆衍生于7q-的丢失。     

伴7号染色体异常的急性白血病32例分析

目的探讨7号染色体异常在急性白血病中的发生率及预后意义。方法采用R带常规显带技术进行染色体检查,对410例急性白血病患者的核型进行分析。结果410例急性白血病患者中检出7号染色体异常患者32例,占7.8%;其中-7/7q-19例(59.4%),t(7;11)3例(9.4%),其他为少见的7号染色体改变der(7), 7,t(2;7),t(5;7),t(7;9),t(7;8),dic(1;7)。-7/7q-中,急性髓细胞白血病12例,其中M0、M1、M2型的-7/7q-发生率高于其他类型急性髓细胞白血病。32例患者中20例同时有其他染色体异常,最常见的是t(9;22)伴-7, 8,-5。30例进行正规化疗的患者,11例缓解,缓解率低于同期急性白血病的总缓解率(36.7%vs65.8%);伴-7/7q-的急性髓细胞白血病的缓解率低于染色体正常的急性髓细胞白血病患者(25%vs55.6%);伴-7/7q-的急性淋巴细胞白血病的缓解率与染色体正常的急性淋巴细胞白血病患者无差异(57.1%vs77.8%),但缓解的4例急性淋巴细胞白血病患者均于短期内复发。伴其他7号染色体异常的11例患者仅4例缓解。结论-7/7q-是7号染色体异常中最为常见的核型改变,且多见于急性髓细胞白血病的M0、M1、M2型,并可能与急性白血病的发病有关;伴7号染色体异常的急性白血病患者预后较差。     

荧光原位杂交技术在骨髓增生异常综合征患者5号和7号染色体异常检测中的应用

目的评价荧光原位杂交（FISH）术在骨髓增生异常综合征（MDS）患者5号和7号染色体异常检测中的应用。方法 40例MDS患者,采用间期FISH,选取4种探针组合检测-5/5q-和-7/7q-。结果利用FISH技术,5号、7号染色体异常检出率分别为15.0%、20.0%。结论 FISH技术可以用于骨髓增生异常综合征（MDS）患者5号和7号染色体异常的检测,采用组合探针的FISH更为敏感和特异。     

间期荧光原位杂交对骨髓增生异常综合征患者5号和7号染色体异常的检测

目的筛选经济、实用的探针组合提高MDS染色体异常的检出率；比较常规细胞遗传学分析（CCA）及荧光原位杂交（FISH）两种技术在MDS染色体异常检测中的灵敏度和特异性。方法采用CCA法和HSH法分析48例患者骨髓细胞的染色体异常情况。结果CCA检出染色体异常18例（37．5％），其中复杂异常4例（8．3％），＋8异常8例（16．6％）、-5／5q-异常5例（10.4％），-7／7q-异常5例（10.4％）、20q-异常2例（4．6％）、不一致的易位3例（6．2％）。FISH除证实CCA发现的-5／5q-和-7／7q-各5例外，还检出2例有5q-，5例有7q-，1例有-7，从而使-5／5q-和-7／7q-的检出率分别增至14．5％和22．9％。平均随访12个月，38例存活，10例死亡，5例转变为急性白血病。结论CCA结合FISH能提高MDS染色体异常的检出率，与CCA相比，采用组合探针的HSH更为敏感和特异。     

荧光原位杂交技术对骨髓增生异常综合征患者染色体异常的检测分析

目的:分析荧光原位杂交(FISH)技术对骨髓增生异常综合征(MDS)5、7、8、20号染色体异常检出率,并与常规细胞遗传学分析(CCA)进行比较。方法:收集48例MDS患者骨髓标本,CCA采用骨髓短期培养法及G带核型分析。FISH采用间期FISH,选取针对5、7、8、20号染色体的不同探针组合。χ2检验比较两种方法检出率。结果:CCA对5、7、8、20号染色体异常检出率分别为8.33%、12.50%、6.25%、12.51%,总的异常检出率为39.58%;FISH对5、7、8、20号染色体异常检出率分别为12.50%、16.67%、10.42%、16.67%,总的异常检出率为56.25%。两法检出率差异无统计学意义(P>0.05),但FISH检测时间(2～3天)明显短于CCA(10～14天)。结论:FISH检测MDS患者5、7、8、20号染色体异常是CCA的重要补充,并明显缩短检测时间。     

158例月经异常患者与染色体异常的关系

目的对月经异常患者行细胞遗传学分析,探讨月经异常与染色体异常的关系。方法用外周血淋巴细胞培养进行染色体制备由2人以上行染色体分析,细胞计数30个,分析3个。结果158例月经异常患者中共检出异常核型33例,异常率20．9％（33／158）。其中纯合型45,X12例,占总数的7．6％,占异常核型的36．4％,嵌合体9例,占总数的5．7％,占异常核型的27．3％,46,XY7例,占总数的4．4％,占异常核型的21．2％,X染色体结构异常5例,占总数的3．2％。占异常核型的15．2％,其它染色体平衡易位1例,占总数的0．6％,占异常核型的3．0％。结论月经异常与性染色体异常密切相关,对月经异常尤其是闭经患者应进行外周血染色体检查。     

C7前结节和第1肋颈间异常韧带的观测及临床意义

目的观察C7横突前结节与第1肋颈间异常韧带与C8颈神经前支的关系,为临床诊治提供解剖学基础.方法32具成人尸体标本,解剖观察C7横突前结节与第1肋颈间异常韧带及其与第1肋颈上面及中小斜角肌内缘所围成的三角形结构与C8颈神经前支的关系.结果C7横突前结节与第1肋颈间异常韧带的出现率为15.63%,其长度为(0.98±0.18) cm,宽度为(0.39±0.11) cm,厚度为(0.06±0.02) cm.C8颈神经前支出椎间孔后经此异常韧带的深面进入三角形结构内,然后跨过中小斜角肌的内缘进入斜角肌间隙.结论C8颈神经前支经过此三角形结构时周围结构的压迫是导致胸廓出口综合征的原因之一.     

C_7前结节和第1肋颈间异常韧带的观测及临床意义

目的 :观察C7横突前结节与第 1肋颈间异常韧带与C8颈神经前支的关系 ,为临床诊治提供解剖学基础。方法 :3 2具成人尸体标本 ,解剖观察C7横突前结节与第 1肋颈间异常韧带及其与第 1肋颈上面及中小斜角肌内缘所围成的三角形结构与C8颈神经前支的关系。结果 :C7横突前结节与第 1肋颈间异常韧带的出现率为 15 .63 % ,其长度为 ( 0 .98± 0 .18)cm ,宽度为 ( 0 .3 9± 0 .11)cm ,厚度为 ( 0 .0 6±0 .0 2 )cm。C8颈神经前支出椎间孔后经此异常韧带的深面进入三角形结构内 ,然后跨过中小斜角肌的内缘进入斜角肌间隙。结论 :C8颈神经前支经过此三角形结构时周围结构的压迫是导致胸廓出口综合征的原因之一。     

应用荧光原位杂交技术验证骨髓增生异常综合征的非克隆性染色体异常

目的探讨应用荧光原位杂交(FISH)技术验证骨髓增生异常综合征(MDS)经传统染色体显带分析(CBA)检测到的非克隆性染色体异常(n-CCA)的价值。方法回顾性分析2011年10月至2020年12月北京大学人民医院CBA检测具有n-CCA的91例MDS患者的临床资料及染色体核型和FISH检测结果。结果在91例患者中, CBA共检测到94例次非克隆性+8、5q-、-7、7q-和20q-异常, 其中43例次(45.7%)经FISH验证为阳性, +8、5q-、-7、7q-和20q-的验证率分别为47.6%(30/63)、25%(2/8)、41.7%(5/12)、40%(2/5)和66.7%(4/6), FISH阳性的细胞占比为4% ～ 90%(中位7%)。将91例患者按CBA分析的中期分裂相数目分为3组, 即≥ 20、10 ～ <20、< 10。3组的FISH验证率分别为43.7%(31/71)、33.3%(3/9)、63.6%(7/11), 未见明显差异(P>0.05)。对26例仅接受支持治疗的患者进行连续监测, 结果显示91.7%(11/12)经FISH验证的异常持续存...     

骨髓增生异常综合征免疫异常的表型

本文探讨免疫异常表型对骨髓增生异常综合征 (MDS )分型诊断及鉴别诊断价值。选用多种单克隆抗体 ,用流式细胞测定分析仪 ,对 4 9例MDS、 5 0例良性血液病患者 (贫血、粒细胞减少症、血小板减少性紫癜、感染等 )的骨髓细胞免疫表型进行测定分析研究。结果MDS 89 3%以上有二系或三系免疫表型异常 ,其中CD7、CD19、CD13、CD14、CD33、CD34、HLA DR免疫标志变化最大 ,明显高于正常对照组 (P <0 0 1) ;髓系抗原表达明显增高 ,而且随MDS进展恶化 ,FAB亚型抗原表达出现规律性变化 ,RA→RAS→RAEB→向RAEB T转化 ,较早期髓系抗原表达 (如CD13、CD33)逐渐增加 ,而较晚期髓系抗原表达 (如CD15、CD14 )逐渐减少 ;同时伴淋系抗原表达逐渐减少 ;骨髓干细胞 /祖细胞表面抗原 (如CD34、HLA DR ) ,随着MDS恶化发展 ,有逐渐明显增加异常表现 ,而且CD34、HLA DR早期抗原表达增高者 ,常常预后较差 ,易于转化成白血病。结论MDS患者骨髓细胞免疫异常表型 ,有利于MDS诊断、分型诊断及鉴别诊断 ,并对治疗、预后判断有重要指导价值[1,2 ,5] 。     

组合探针荧光原位杂交检测骨髓增生异常综合征常见染色体异常

目的：评价组合探针荧光原位杂交（fluorescence in site hybridization,FISH)在检测骨髓增生异常综合征（myelodysplastic syndrome,MDS）常见染色体异常中的价值。方法：应用YAC248F5（5q31)、YAC938G5（7q32）、CEP8、YAC912C3（20q12）4种DNA探针，对核型未知的20例MDS患者进行FISH检测－5／5q-、－7／7q-、＋8、20q－等常见染色体异常，并与常规细胞遗传学分析结果相比较。结果：20例MDS患者中，组合探针FISH检出13例有常见染色体异常（其中5例＋8，1例－5／5q-，5例20q-,1例5q-合并20q-，复杂异常1例）；而常规细胞遗传学发现5例常见染色体异常，1例＋21，复杂异常1例，标记染色体1例，正常5例。结论：组合探针FISH是筛查MDS患者常见染色体异常的有效手段。     

Diagnostic yield of bone marrow and peripheral blood FISH panel testing in clinically suspected myelodysplastic syndromes and/or acute myeloid leukemia: a prospective analysis of 433 cases

It is unclear how often and in what setting fluorescence in situ hybridization (FISH) panels for myelodysplastic syndrome (MDS)/acute myeloid leukemia (AML) provide additional information over metaphase cytogenetics alone. Furthermore, the usefulness of peripheral blood vs bone marrow FISH has also not been directly compared. We prospectively compared metaphase cytogenetics and FISH for -5/5q-, -7/7q-, +8, and 20q- in 433 cases of suspected MDS/AML. FISH testing was abnormal in 6 (14%) of 43 and 10 (19%) of 54 cases with fewer than 20 normal metaphases or no growth, respectively. FISH was only rarely abnormal in cases with 20 normal metaphases obtained (6/222 [2.7%]). Comparison of peripheral blood and bone marrow results in 48 cases showed abnormal peripheral blood FISH results in 18 (69%) of 26 cases with abnormal bone marrow FISH results and in 5 (23%) of 22 cases with normal bone marrow FISH results. These findings, the largest published comparison of FISH vs metaphase cytogenetics in MDS/AML, provide a rational strategy for FISH testing in peripheral blood and bone marrow.     

恶性血液病8号染色体数目异常的间期荧光原位杂交检测

目的　探讨间期荧光原位杂交 (fluorescenceinsituhybridization ,FISH)技术在检测恶性血液病 8号染色体数目异常中的价值。方法　采用常规细胞遗传学 (conventionalcytogenetics ,CC)和 8号染色体着丝粒特异性探针间期FISH技术对 8例CC检测显示 8号染色体数目异常的急性髓细胞样白血病患者、10例慢性髓细胞样白血病加速期或急变期患者和 3名正常人骨髓进行 8号染色体数目检测。结果9例CC检测为三体 8的患者中 ,FISH检测结果均与其一致 ,其中例 5经CC检测仅发现存在二体 8、三体8和四体 8克隆 ,而FISH检测不但证实了三体 8和四体 8克隆的存在 ,还发现存在一个较小的五体 8克隆。例 3和例 17经CC检测只发现一个细胞有三体 8,无法确定是否为三体 8克隆性畸变 ,FISH检测证实有三体 8克隆存在。例 9经CC检测未发现三体 8,FISH检测发现有三体 8克隆存在。与CC检测结果相比 ,除例 16三体 8检出率FISH结果明显高于CC检测结果外 ,其余均低于或接近CC检测结果。结论　间期FISH技术对检测 8号染色体数目异常具有重要价值 ,当CC检测正常、不肯定或中期分裂相质量差、数量少时作用更大 ,是CC的重要补充。     

Acquisition of a Ph chromosome with minor BCR/ABL fusion in treatment-related myelodysplastic syndrome with chromosome 7 abnormalities in a patient treated for Hodgkin disease

The patient reported in this study originally had Hodgkin disease that was treated heavily with multiple courses of combined chemotherapy and radiotherapy. Secondary myelodysplastic syndrome (MDS) with a complex karyotype with monosomy 7, deletion 7q31, and double deletion 7q31 developed 8 years later. During the course of the disease, conventional cytogenetics and interphase FISH (I-FISH) analysis detected a Ph chromosome and BCR/ABL fusion with mBCR rearrangement. Using a multiparametric cell scanning system that enables combined analysis with probes specific for 7/7q- and BCR/ABL in a single cell, we were able to demonstrate the presence of the BCR/ABL fusion only in cells with monosomy of chromosome 7 and 7q31 deletion, but not in cells with a normal chromosome 7 or with a double deletion of 7q31. We propose two possible models that may explain the appearance of the BCR/ABL fusion in the pre-existing treatment-related MDS clones characterized by chromosome 7 rearrangements.     

Trisomy 8 and monosomy 7 detected in bone marrow using primed in situ labeling, fluorescence in situ hybridization, and conventional cytogenetic analyses. A study of 54 cases with hematological disorders

Trisomy 8 and monosomy 7 are the two most frequent aneuploidies found in hematological disorders such as myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML). In this study, primed in situ labeling (PRINS), fluorescence in situ hybridization (FISH) and conventional cytogenetic approaches were used to investigate 54 cases of hematopoietic disorders. Of these cases, there were 22 cases of trisomy 8, 2 cases of tetrasomy 8, 14 cases of monosomy 7, and 16 cases with two copies of both chromosomes 7 and 8. PRINS was carried out in interphase nuclei of bone marrow samples using primers that can specifically detect alpha-satellite DNA sequences of chromosomes 7 and 8. In parallel, using the alpha-satellite probes for chromosomes 7 and 8, FISH was performed for all the cases. PRINS and FISH techniques showed similar specificity and sensitivity. In comparison to FISH, PRINS is more advantageous since it is a faster, easier, and more cost-effective technique. Additionally, for most of the cases, a higher proportion of aneuploidy was detected in metaphases using conventional cytogenetics, as compared to the one found in interphase nuclei identified with PRINS and FISH techniques. In other words, for these cases, the cells with trisomy 8 or monosomy 7, had a distinct proliferative advantage compared to the disomic cell population. Therefore, to better quantify the proportion of aneuploid cells in bone marrow, we recommend to investigate the frequency of aneuploidy in nuclei using PRINS, rather than studying only the dividing cells as detected by conventional cytogenetic techniques.     

Detection of monosomy 7 by fluorescenceIn situ hybridization in acute nonlymphocytic leukemia and myelodysplastic syndrome

Summary Fluorescencein situ hybridization (FISH) with a chromosome 7 specific alpha satellite DNA probe was used to detect monosomy 7 in interphase and metaphase cells obtained from patients with myelodysplastic syndrome (MDS) and acute nonlymphocytic leukemia (ANLL). Chromosome analysis revealed monosomy 7, either alone or as part of a complex chromosome abnormality, in all cell samples. FISH analyses of 12 marrow samples and a blood sample using a chromosome 7 specific alpha satellite DNA probe revealed a single fluorescence spot in 80.5–97.5% of interphase cells indicating monosomy 7. In contrast, 83.5–92.0% of the same cells had two copies of chromosome 17 as two fluorescent spots were detected using a chromosome 17 specific alpha satellite DNA probe used as a positive control. The proportion of interphase cells with monosomy 7 did not correlated with the percentage of metaphase cells with monosomy 7 detected by conventional karyotyping or with the percentage of blast cells in the bone marrow.     

Molecular cytogenetic profiling of complex karyotypes in primary myelodysplastic syndromes and acute myeloid leukemia

Complex chromosomal aberrations are present in < or =30% of patients with primary myelodysplastic syndrome (MDS) or acute myeloid leukemia (AML) and are associated with a poor prognosis. Specific alterations in complex karyotypes are difficult to define by conventional cytogenetics alone. To obtain a more comprehensive view of the recurrent aberrations, we performed spectral karyotyping (SKY) and fluorescence in situ hybridization (FISH) with selected probes on bone marrow samples from 17 patients with primary MDS and 3 with primary AML. All cases had chromosome 5 alterations. Two different types of 5q loss were identified: unbalanced translocations and interstitial deletions, or del(5q), each occurring in 10 patients. The most frequent additional chromosome aberrations were -3/-3p/-3q, -7/7q-, +8, 13q-, -16, 17p-, -18/18p-, -20/20q-, and +21q, each occurring in 25%. In the five cases with gain of 21q, involvement of the AML1 gene was excluded. Unbalanced 5q translocations occurred more often in combination with monosomy 3 and 7 and with gain of 21q, whereas del(5q) was associated more often with -1p and trisomy 8. A detailed analysis of specific breakpoints and deletions revealed recurrent involvement of specific chromosomal bands harboring known tumor suppressor genes or oncogenes. Analysis of large numbers of MDS and AML cases in a similar detailed manner with SKY and FISH will reveal whether new subgroups can be identified according to their genetic alterations. Correlation with clinical parameters may reveal the prognostic significance of these genetic subgroups.