荧光原位杂交检测多发性骨髓瘤患者13号染色体缺失和IgH基因易位

目的分析多发性骨髓瘤（MM）患者13号染色体长臂缺失[del（13q）]和免疫球蛋白重链（IgH）基因易位[t（14q）]与临床相关因素的关系。方法采用间期荧光原位杂交（FISH）技术对25例初诊MM患者应用探针D13S319、IgH分别进行del（13q）和t（14q）的检测。结果 25例患者中del（13 q）8例（32.0%）、IgH易位4例（16.0%）,同时存在上述2种异常2例（8.0%）。伴del（13 q）及IgH易位的患者具有较高的血清β2-微球蛋白（β2-MG）水平及较高的骨髓浆细胞百分比。结论间期FISH是一种检测MM患者骨髓瘤细胞突变的敏感方法,具有临床应用价值。del（13q）、t（14q）对判断临床疗效和预后具有指导作用。     

多发性骨髓瘤分子细胞遗传学异常的研究

目的探讨多发性骨髓瘤（MM）的分子细胞遗传学异常。方法应用CD138单克隆抗体磁珠分选系统纯化23例初治MM患者的骨髓浆细胞，结合一组探针和间期荧光原位杂交技术检测MM患者13q14缺失、p53缺失以及IgH基因重排的发生率。结果23例MM患者中，10例（43．5％）13q14缺失。阳性率为79％-96％；11例（47．8％）IgH基因重排；7例（30．4％）有13q14缺失和IgH基因重排；所有病例均未检测到p53基因缺失。结论13q14缺失及IgH基因重排在MM患者中的发生率较高；13q14的缺失和IgH基因重排的发生率同疾病进展、预后的关系有待进一步研究。     

FICTION技术在检测多发性骨髓瘤遗传学异常中的应用

目的 探讨联合免疫荧光和荧光原位杂交(FISH)的FICTION技术在多发性骨髓瘤(MM)遗传学异常检测中的应用价值.方法 采集18例MM患者和2例浆细胞白血病患者的骨髓标本,分离单个核细胞制作滴片.从细菌人工染色体文库中选取覆盖IgH、MMSET待测基因位点的质粒,用缺口平移法制备带有半抗原检测标签的核酸探针.在经CD138标记和酪胺信号放大的细胞滴片标本上,使用上述自制探针[t(4;14)、t(11;14)和t(14;16)]和商品化直标缺失探针(13q和p53)进行FISH检测.结果 20例患者标本均使用上述5种探针进行检测,其中检出t(4;14)4例,t(11;14)6例,t(14;16)1例,p53缺失3例,13q缺失8例;另有4例未检测出此5种异常.结论 应用FICTION技术原位分析骨髓中特定瘤细胞亚群的特征性遗传学异常,能够提高FISH检测的效率和敏感性,并可作为对MM患者遗传学分层诊断的初筛实验,指导治疗并判断预后.

Abstract：

Objective To investigate the diagnostic value of FICTION (Fluorescence Immunophenotyping and Interphase Cytogenetics as a Tool for the Investigation of Neoplasms) technique, combining immunofluorescence and fluorescence in situ hybridization (FISH), to detect genetic aberrations in multiple myeloma (MM). Methods Bone marrow samples were collected from 18 MM and 2 plasma cell leukemia (PCL)patients. Probes targeting IgH and MMSET were prepared using a Nick Translation Kit from Bacterial artificial chromosome (BAC) clones. The immunophenotyping was achieved via the CD138 tyramide signal amplification (TSA)-mediated immunofluorescence, followed by FISH with the prepared probes [t (4;14), t (11;14), t(14;16)] and the commercial deletion probes (13q and p53) to detect common genetic aberrations in MM. Results All the 20 samples were assayed with the probes mentioned above, and revealed 4 cases with t(4;14) ,6 with t(11 ;14), 1 with t(14;16), 3 with p53 deletion; and 8 with 13q deletion. The remaining 4 cases had none of the 5 aberrations. Conclusion FICTION technique facilitates the detection of genetic abnormalities of MM in situ; enhances both efficiency and sensitivity of positive det~tion, thus, could be used as the screening test of molecular diagnosis of MM to guide coming-up risk-adapted therapy and evaluate prognosis.     

FICTION技术在检测多发性骨髓瘤遗传学异常中的应用

目的 探讨联合免疫荧光和荧光原位杂交(FISH)的FICTION技术在多发性骨髓瘤(MM)遗传学异常检测中的应用价值.方法 采集18例MM患者和2例浆细胞白血病患者的骨髓标本,分离单个核细胞制作滴片.从细菌人工染色体文库中选取覆盖IgH、MMSET待测基因位点的质粒,用缺口平移法制备带有半抗原检测标签的核酸探针.在经CD138标记和酪胺信号放大的细胞滴片标本上,使用上述自制探针[t(4;14)、t(11;14)和t(14;16)]和商品化直标缺失探针(13q和p53)进行FISH检测.结果 20例患者标本均使用上述5种探针进行检测,其中检出t(4;14)4例,t(11;14)6例,t(14;16)1例,p53缺失3例,13q缺失8例;另有4例未检测出此5种异常.结论 应用FICTION技术原位分析骨髓中特定瘤细胞亚群的特征性遗传学异常,能够提高FISH检测的效率和敏感性,并可作为对MM患者遗传学分层诊断的初筛实验,指导治疗并判断预后.     

FICTION技术在检测多发性骨髓瘤遗传学异常中的应用

目的 探讨联合免疫荧光和荧光原位杂交(FISH)的FICTION技术在多发性骨髓瘤(MM)遗传学异常检测中的应用价值.方法 采集18例MM患者和2例浆细胞白血病患者的骨髓标本,分离单个核细胞制作滴片.从细菌人工染色体文库中选取覆盖IgH、MMSET待测基因位点的质粒,用缺口平移法制备带有半抗原检测标签的核酸探针.在经CD138标记和酪胺信号放大的细胞滴片标本上,使用上述自制探针[t(4;14)、t(11;14)和t(14;16)]和商品化直标缺失探针(13q和p53)进行FISH检测.结果 20例患者标本均使用上述5种探针进行检测,其中检出t(4;14)4例,t(11;14)6例,t(14;16)1例,p53缺失3例,13q缺失8例;另有4例未检测出此5种异常.结论 应用FICTION技术原位分析骨髓中特定瘤细胞亚群的特征性遗传学异常,能够提高FISH检测的效率和敏感性,并可作为对MM患者遗传学分层诊断的初筛实验,指导治疗并判断预后.     

人多发性骨髓瘤细胞系与初治多发性骨髓瘤遗传学异常的比较研究

本研究采用荧光原位杂交（fluorescence in situ hybridization,FISH）检测人多发性骨髓瘤细胞系（human multiple myeloma cell lines,HMCL）和初治多发性骨髓瘤（MM）的分子遗传学异常并进行比较分析,以帮助筛选浆细胞肿瘤高危遗传学异常,为多发性骨髓瘤MM的危险度分层提供依据,并为利用HMCL进行MM研究提供遗传学资料。选用13q14（RB-1）、14q32（IGHC/IGHV）、1q12（CEP1）、17p13（TP53）荧光探针应用直接法检测7株HMCL,用CD138免疫磁珠分选（magnetic-activated cell sorting,MACS）联合FISH方法检测85例初治MM患者作为对照。对于存在14q32异常的患者进行IGH/CCND1、IGH/FGFR3、IGH/MAF双色双融合探针杂交以检测t（11;14）（q13;q32）、t（4;14）（p16;q32）、（14;16）（q32;q23）。结果显示：7株HMCL中RB-1/D13S19缺失6株（85.7%）,P53缺失5株（71%）,1q21扩增6株（85.7%）;5株（71.4%）HMCL存在IGH异常,IGH/CCND1阳性细胞系1株,IGH/FGFR3阳性细胞系2株,IGH/MAF阳性细胞系3株;其中KMS11细胞系同时存在IGH/FGFR3和IGH/MAF两种异常。85例初治MM细胞遗传学总体检出率为85.9%,38例（44.7%）伴有RB-1缺失,17例（20%）出现p53缺失,45例（52.9%）伴有1q21扩增,53例（62.4%）存在IGH异常,其中IGH/CCND1阳性23例（27.1%）,IGH/FGFR3阳性21例（24.7%）,IGH/MAF阳性3例（3.5%）。与初治MM相比较,HMCL的抑癌基因p53缺失率和IGH/MAF明显升高（p=0.008,p=0.005）,而其他分子遗传学异常检出率未达统计学差异。结论：HMCL作为浆细胞肿瘤恶性程度最高的形式,积累了大量的遗传学异常,p53缺失是其显著特征。绝大多数HMCL来源于疾病晚期的髓外浆细胞肿瘤细胞或者浆细胞白血病,提示p53缺失是这部分极高危浆细胞肿瘤患者的特征。     

染色体13q14缺失在多发性骨髓瘤发生发展中的意义及对其临床预后的影响

目的 研究染色体13q14缺失在多发性骨髓瘤(MM)患者中的发生率及其临床意义.方法 对我院淋巴瘤中心132例初治MM患者骨髓标本行CD138免疫磁珠富集骨髓瘤细胞后,采用13q14(RB1)探针进行荧光原位杂交(FISH)检测,结合不同治疗方案分析其临床意义.结果 ①检出率:FISH检测13q14缺失率为51.5%,而常规染色体核型分析△l3(-13/13q-)检出率仅为5.0%.②单因素分析显示,13q14缺失比例＞25%、ISS分期为Ⅱ或Ⅲ期、血β2-MG≥5.5 mg/L、起病时骨髓涂片浆细胞比例＞0.500是不良预后因素.多因素分析显示,只有13q14缺失比例＞25%是独立的不良预后因素.硼替佐米与传统化疗相比可明显提高13q14缺失患者的近期疗效.应用硼替佐米后13q14缺失比例＞25%患者与缺失比例≤25%患者的总生存时间差异无统计学意义,提示硼替佐米能克服13q14缺失对预后的不良影响.结论 CD138磁珠分选后行FISH检测可以显著提高MM患者中13q14缺失的检出率;FISH检测MM患者13q14缺失比例＞25%是独立的预后不良因素,且与患者自身的肿瘤负荷、其他遗传学指标密切相关;硼替佐米可以克服13q14缺失对近期疗效的不良影响.

Abstract：

Objective To determine the incidence and clinical significance of chromosome 13q14 deletion in multiple myeloma(MM). Methods Bone marrow samples were collected from 132 newly diagnosed MM patients referred to our hospital. Interphase fluorescence in situ hybridization (i-FISH) combined ith magnetic activated cell sorting (MACS) were performed on chromosome 13q14(RB-1). Results ①i-FISH was used to investigate CD138-enriched bone marrow MM cells and revealed a 13q14 deletion rate of 51.5% (68/132), while conventional cytogenetic (CC) analysis revealed 13q deletions/monosony13 (△13)only of 5.0% (6/120). ②Univariate analysis showed that 13q14 deletion rate by i-FISH ＞25%, bone marrow plasma cells ＞ 50%, ISS stage and β2 -MG ≥ 5.5 mg/L were associated with shorter overall survival (OS). Multivariate analysis revealed that 13q14 deletion rate by i-FISH ＞ 25% was an independent unfavorable factor (P = 0.042). ③Patients treated with bortezomib had a much better response than those treated with traditional chemotherapy (P = 0. 001). There was no significant difference in OS between patients received bortezomib with and without 13q14 deletion (P ＞0.05), indicating that bortezomib could reverse the poor prognosis of 13q14 deletion. Conclusion ①i-FISH followed CD138 cell sorting appeares to be a highly sensitive method for detecting 13q14 deletion. ②13q14 deletion rate by i-FISH ＞25% is an independent unfavorable factor. ③Bortezomib could reverse the poor prognosis of l3q14 deletion.     

伴有14q32异常的淋巴系统恶性肿瘤的临床与细胞遗传学分析

目的 分析14q32异常的淋巴系统恶性肿瘤（lymphiod maligances,LM)的临床及细胞遗传学特征。方法 应用常规细胞遗传学技术对225例不同类型的LM进行研究并综合分析伴14q32异常LM的临床及实验室特征。结果 15（6.67%）LM患者可见14q32异常,不同类型的14q32异常的分布与LM的不同类型有关。t（8;14）（q24;q32)）最为常见,主要见于急性白血病,其白血病类型呈现形态学及免疫表型特征的异质性,但仍具有独特的临床、预后特征和附加的染色体异常ins（1;6）(q11;q23q27)。t(11;14）（q13;q32)仅见于浆细胞白血病,1例多发性骨髓瘤继发骨髓增生异常综合征（MDS）患者可见累及MDS常见的染色体异常7q-和20q-。结论 特征性14q32异常与附加染色体改变有助于同类型的LM的诊断和预后估计。     

检测骨髓异常浆细胞有助于诊断原发性系统性轻链淀粉样变性

原发性系统性轻链淀粉样变性(AL)是系统性淀粉样变性疾病中最常见的类型,最近已明确AL是异常浆细胞导致的恶性肿瘤。这些浆细胞产生过量的免疫球蛋白轻链,形成具有蛋白质毒性的纤维状淀粉样物,通过异常折叠和沉积于多种组织器官,引起多脏器损害。AL诊断较困难,以往主要通过活检组织的刚果红染色。现在使用流式细胞术可以通过患者骨髓浆细胞的免疫球蛋白轻链限制性及异常表型特征检测异常浆细胞克隆,有助于早期确诊AL,并作为诊断AL后临床疗效监测的重要指标。本文就AL的诊断和分型、临床特点、骨髓浆细胞免疫表型的流式细胞术检测,异常浆细胞免疫球蛋白轻链限制性和表型特点等作一简要的综述。     

CD5-negative blastoid variant mantle cell lymphoma with complex CCND1/IGH and MYC aberrations

The coexistence of CCND1/IGH and MYC rearrangements in mantle cell lymphoma (MCL) is a rare finding associated with a very poor prognosis. In this study, a patient with blastoid variant (MCL) is reported. The disease was clinically aggressive and refractory to chemotherapy, and the patient only survived for 1 month following diagnosis. Conventional cytogenetic study, FISH, and multicolor FISH (mFISH) demonstrated the involvement of the BCL1/CCND1 locus in a complex translocation, t(3;11)(q25;p15)t(11;14)(q13;q32). In addition, subclonal abnormalities in the 8q24 region, manifested as a t(8;14)(q24;q32)/MYC rearrangement, were identified. To the best of our knowledge, this is the first MCL case in Korea bearing these complex genomic aberrations.     

Detection of translocations diagnostic for Berkitt's lymphoma by fluorescent in situ hybridization on histological sections of paraffin blocks

AIM: To test feasibility of detection of translocations which are diagnostic for Berkitt's lymphoma with the method of fluorescence in situ hibridization (FISH) on histological sections of paraffin blocks. MATERIAL AND METHODS: FISH on histological sections for detection of t(8;14)(q24;q32) and variant t(2;8)(p12;q24) and t(8;22)(q24;q11) was performed on the material obtained from 53 patients with typical clinical, morphological and immunological picture. DNA probe LSI IgH/MYC, CEP 8 Tri-color, Dual Fusion Translocation Probe (Vysis, USA), for variant translocations DNA-probe LSI MYC, Dual color, Break Apart Rearrangement Probe (Vysis, USA) were used. RESULTS: Histological material from 31 patients contained translocations characteristic for LB: in 29 (93.5%)--t(8;14)(q24;q32), in 2--variant rearrangements of locus of gene c-myc. Translocation t(8;14)(q24;q32) and its variants were not detected in 22 patients, the diagnosis was changed for diffuse large B-cell lymphoma (DLBCL). CONCLUSION: Typical for BL clinical, morphological and immunological picture may present in extra-nodal diffuse large B-cell lymphoma with high proliferative activity. Differential diagnosis between BL and the latter lymphoma is possible only basing on detection of translocation t(8;14)(q24;q32) or its variants. If it is impossible to obtain native material, FISH on histological sections of parasffin blocks is the only possible method of differential diagnosis.     

Cytogenetic and molecular basis of multiple myeloma

Multiple myeloma(MM) originating from plasma cells is characterized by complex chromosomal aberrations. The most prominent chromosomal abnormalities of MM are aneuploidy and translocations affecting the immunoglobulin heavy chain locus on chromosome 14q32. Additionally, a variety of genetic aberrations such as ras mutations have been found in MM. Because these chromosomal and genetic abnormalities are closely associated with clinical behavior including prognosis, cytogenetic findings have a great impact on planning treatment strategy. Furthermore, studies of signal transductions and mechanisms of oncogenesis in association with these abnormalities will provide targets to develop novel therapeutic agents. Here we summarize the chromosomal and genetic abnormalities of MM and their clinical implications.     

8号染色体四体、三体共存的t(15;17)(q22;q12)急性早幼粒细胞白血病

本研究报道首例伴有8号染色体四体(四体8)、8号染色体三体(三体8)异常的t(15；17)急性早幼粒白血病(AML-M3a)，并探讨其形态学、细胞遗传学、分子生物学、免疫学及临床特点。用外周血及骨髓标本直接涂片观察形态学改变；采用骨髓细胞24小时短期培养法制备染色体标本，RHG显带技术进行核型分析；以筑巢式逆转录聚合酶链反应(nested RT—PCR)技术检测PML-RARa融合基因转录本；以间期荧光原位杂交(fluorescence in situ hybridization，FISH)技术检测8号染色体数目异常；以流式细胞术检测免疫表型。结果表明：外周血涂片早幼粒细胞占65％，可见中晚幼粒细胞。骨髓涂片显示有核细胞增生明显活跃，粒系83．6％，其中早幼粒细胞占72．4％，胞浆内可见大量紫红色颗粒。染色体核型分析揭示核型为48，XY， 8， 8，t(15；17)(q22；q12)[16]／47，XY， 8，t(15；17)(q22；q12)[3]／46，XY，t(15；17)(q22；q12)[1]。RT—PCR检测PML-RARa( )。FISH检测显示具有1，2，3，4，5，6个绿色荧光信号细胞的百分比分别为0．5，7，19，55，18和0．5。这不但证实了三体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。白血病细胞免疫表型检测显示CD13(96．2％)、CD33(55．9％)、CYMPO(93．5％)阳性，其余抗原包括淋系抗原在内均为阴性。患者生存期只有10天。结论 本例四体8是t(15；17)的继发性改变，可能是三体8克隆进展的结果。伴有四体8的t(15；17)AML-M3预后差。     

Overexpression of G protein-coupled receptor 5D in the bone marrow is associated with poor prognosis in patients with multiple myeloma

Eur J Clin Invest 2012; 42 (9): 953-960 ABSTRACT: Background G protein-coupled receptor 5D (GPRC5D) is a novel surface receptor. As this new subtype of G protein-coupled receptors was discovered, little is known about the role of this gene. Materials and methods In this retrospective study, we investigated GPRC5D mRNA expression by real-time polymerase chain reaction (RT-PCR) in bone marrow (BM) of 48 patients with multiple myeloma (MM). Results Highly variable levels of GPRC5D (median, 288; quartiles, 17-928) were detected in patients with MM, whereas only low expression was detected in normal tissues (median, 1; quartiles, 1-23). High mRNA expression of GPRC5D correlated positively with high plasma cell count in bone marrow (r?=?0·64, P?相似文献   

免疫磁珠分选对荧光原位杂交检测多发性骨髓瘤细胞遗传学异常的影响

本研究旨在明确免疫磁珠浆细胞分选对使用荧光原位杂交(FISH)技术检测多发性骨髓瘤(MM)细胞分子遗传学异常的影响,探索适合我国国情的检测方法。对同一MM患者的标本进行2种方法检测:1种为不经分选,直接进行FISH;另1种为经过CD138免疫磁珠分选(MACS)富集浆细胞后进行FISH检测;比较2种FISH技术对MM遗传学异常检出率并分析其影响因素。探针类型为13q14(RB-1)和14q32(IGH)。结果表明:①利用13q14(RB-1)探针对42例患者同时进行了分选和不分选的的FISH检测显示,经过CD138磁珠分选后25例(56.8%)检测到13q14缺失;不经分选直接进行FISH检测仅9例(21.4%)患者检出13q14缺失,2种标本处理方法的检出率具有统计学差异(p=0.01);②利用14q32(IGH)探针对22例患者同时进行了分选和不分选的FISH检测显示,经过CD138磁珠分选后14例(63.6%)可以检测到14q32重排,不经分选直接进行FISH检测仅7例(31.8%)患者检出14q32重排,2种标本处理方法的检出率具有统计学差异(p=0.035);③不分选FISH的阳性细胞检出率与骨髓涂片和流式细胞仪所检测的浆细胞比例呈明显的正相关。当骨髓涂片细胞中浆细胞比例超过50%,或者流式细胞仪检测浆细胞比例超过10%时,不分选FISH与磁珠分选FISH结果之间没有统计学差异。结论:MACS-FISH显著提高了MM遗传学异常的检出率,避免了假阴性的发生;直接进行FISH检测的检出率低的主要原因是检测标本中浆细胞比例较低;未经分选直接进行13q14(RB-1)和14q32(IGH)探针FISH检测,其阳性细胞率与骨髓涂片、流式细胞仪所检测出的浆细胞比例呈显著正相关,说明随着所检验标本中浆细胞比例的升高,不进行分选直接进行FISH检测的缺点得到一定弥补。当骨髓涂片细胞中浆细胞比例超过50%,或者流式细胞仪检测浆细胞比例超过10%时,2种检测方法之间没有差异。     

Low-grade B-Cell lymphomas with plasmacytic differentiation lack PAX5 gene rearrangements

The chromosomal translocation t(9;14)(p13;q32) has been reported in association with lymphoplasmacytic lymphoma (LPL). Although this translocation involving the paired homeobox-5 (PAX5) gene at chromosome band 9p13 and the immunoglobulin heavy chain (IgH) gene at 14q32 has been described in approximately 50% of LPL cases, the actual number of cases studied is quite small. Many of the initial cases associated with t(9;14)(p13;q32) were actually low-grade B-cell lymphomas with plasmacytic differentiation other than LPL. Thus, we analyzed a series of low-grade B-cell lymphomas for PAX5 gene rearrangements. We searched records from the Department of Pathology, Stanford University Medical Center for low-grade B-cell lymphomas, with an emphasis on plasmacytic differentiation, that had available paraffin blocks or frozen tissue. We identified 37 cases, including 13 LPL, 18 marginal zone lymphomas (nodal, extranodal, splenic, and alpha-heavy chain disease), and 6 small lymphocytic lymphomas. A novel dual-color break-apart bacterial artificial chromosome probe was designed to flank the PAX5 gene, spanning previously described PAX5 breakpoints, and samples were analyzed by interphase fluorescence in situ hybridization. All cases failed to demonstrate a PAX5 translocation, indicating that t(9;14)(p13;q32) and other PAX5 translocations are uncommon events in low-grade B-cell lymphomas with plasmacytic differentiation. This study also confirms recent reports that found an absence of PAX5 rearrangements in LPL, suggesting the reassessment of PAX5 rearrangements in LPL.     

伴有t（2；21；8）（p12；q22；q22）急性髓系白血病M2的MICM特征与诊断的探讨

本研究应用形态学、免疫学、细胞遗传学和分子生物学（MICM）分型技术联合检测伴有复杂变异核型t（2;21;8）（p12;q22;q22）的急性髓系白血病（AML—M2）,并探讨其特征及诊断意义。取患者骨髓涂片行瑞氏-姬姆萨染色和细胞化学染色进行骨髓细胞形态学FAB分型;流式细胞术（FCM）检测白血病细胞免疫表型;新鲜骨髓细胞短期培养法常规制备染色体标本,RHG显带技术进行核型分析,采用双色双融合AML／ETO探针及全染色体涂染（CP）探针检测有丝分裂中期荧光原位杂交（FISH）信号,并与常规R显带细胞遗传学检测结果进行比较分析,巢式RT—PCR检测AML1-ETO融合转录本。结果表明：病例1原始粒细胞伴嗜酸粒细胞及单核细胞比例增高;病例2符合以异常中幼粒细胞增高为主的AML—M2b;染色体核型分析结合FISH检测证实两例患者均存在t（2;21;8）（p12;q22;q22）复杂核型;AMLL／ETO融合基因转录本阳性,免疫表型显示CD34和HLA—DR共表达,伴CD19和cD56表达。结论：应用WHO提出的细胞形态学、免疫学、细胞遗传学和分子生物学技术（MICM）联合检测的实验室诊断方法对准确诊断复杂变异核型t（2;21;8）（p12;q22;q22）AML的分型具有重要意义。     

Immunoglobulin heavy chain gene rearrangement in heavy chain deposition disease suggests it is a plasma cell disease: a case report

Heavy chain deposition disease (HCDD) is characterized by the deposition of truncated monoclonal immunoglobulin heavy chains along glomerular basement membranes. Truncated heavy chains are thought to be associated with plasma cell disease (PCD), but previous bone marrow cytology tests showed that only 30% of HCDD cases are related to PCDs. We report the first known use of immunoglobulin heavy chain (IGH) gene rearrangement to diagnose a patient with γ3-HCDD, although bone marrow morphology test identified no abnormalities. Our findings provide strong evidence for a correlation between PCDs and HCDD, which could help understand the genetic background underlying abnormal heavy chains and assess disease prognosis. Further, concordant with previous findings, bortezomib-based chemotherapy had a good therapeutic effect in our patient. We summarize the experience of diagnosing and treating a case of HCDD, and combine this with a literature review to further explore the correlation between PCDs and HCDD, which has important clinical value.     

2-甲氧基雌二醇对原代骨髓瘤细胞分化的作用

本研究观察2-甲氧基雌二醇对多发性骨髓瘤患者原代细胞诱导的分化作用。采用CD138+磁珠分离纯化7例骨髓瘤患者原代骨髓瘤细胞,通过形态观察、细胞表面标志分析和细胞分泌轻链蛋白的情况,观察2-甲氧基雌二醇对多发性骨髓瘤原代骨髓瘤细胞分化的影响。结果表明:患者原代骨髓瘤细胞经0.5μmol/L2-甲氧基雌二醇分别作用36及72小时后,细胞形态向成熟阶段发展,表现为细胞胞核缩小,胞浆丰富,核浆比例下降,核仁减少或消失,核染色质变粗、变密。CD49e阳性细胞率由(11.02±1.75)%(DMSO对照组)增加到(23.80±1.62)%(36小时组)及(35.68±1.85)%(72小时组),差异具有统计学意义(P<0.05);细胞分泌轻链蛋白由(225.7±30.4)ng/ml(DMSO对照组)升高到(296.7±18.8)ng/ml(36小时组)及(378.9±15.8)ng/ml(72小时组),差异具有统计学意义(P<0.05)。结论:较低浓度2-甲氧基雌二醇可诱导骨髓瘤患者原代骨髓瘤细胞向成熟阶段分化。     

Detection of bone marrow infiltration of lymphoma cells by fluorescence in situ hybridization

BACKGROUND: It is sometimes difficult to detect the bone marrow infiltration of lymphoma cells, because lymphoma cells are not distinguishable from normal lymphocytes due to the similarity of their phenotype. METHODS: Bone marrow involvement of 17 samples of 15 patients with follicular lymphoma, whose lymphoma cells were confirmed to harbor the translocation of chromosome14q32, were examined by microscopic analysis of bone marrow smear and biopsy, flow cytometorical analysis (FCM), chromosomal analysis of G-banding and fluorescence in situ hybridization (FISH). FISH was performed using a probe, which detects the split of IGH gene on 14q32. RESULTS: The positivity of FISH was highest among these methods and FISH was able to detect the bone marrow involvement in one case who was defined as negative by bone marrow biopsy. CONCLUSIONS: FISH can be used for detection of bone marrow involvement of malignant lymphoma that carries chromosomal rearrangement involving 14q32.