骨髓增生异常综合征345例病态造血特点分析

 目的　分析骨髓增生异常综合征（MDS）病态造血特点。方法　收集2003年7月4日至2007年3月14日原因不明血常规异常的成年患者标本716例,以WHO MDS分类标准为诊断金标准,分别进行细胞形态学、细胞化学染色、骨髓病理检查、细胞遗传学、流式细胞术等检测。分析骨髓细胞学检查中病态造血特征在判断克隆性和非克隆性疾病中的诊断价值,计算灵敏度和特异度。结果　MDS病态造血形态学诊断的主要依据：粒系Auer小体、核出芽、微核有其中之一者;红系核出芽;外周血片中出现巨核细胞;外周血片中出现原粒细胞或早幼红细胞;环状铁粒幼细胞>1 %。次要依据：粒系假Pelger-Hǔet 异常、不能分叶中性粒细胞、同一细胞内核发育不同步、环形核、核染色质聚集;红系的多核、奇数核、子母核、核碎裂、空泡、成熟红细胞大小悬殊;微巨核。结论　细胞形态学是诊断MDS的基础,但也存在一定的局限性,尤其是对于早期MDS细胞形态学改变不典型时,需要结合其他检测手段分析。     

伴髓系抗原CD13高表达的急性B细胞白血病形态学及流式细胞学特点分析

目的 分析伴异常表达CD13急性B细胞白血病形态学及流式细胞学特点,提高诊断此类疾病的诊断水平.方法 对确诊的4例患者的形态学及流式细胞术散点图进行分析,研究并复习相关文献,总结特点.结果 急性B细胞白血病伴异常表达CD13的患者形态学上多数具有高度异质性与分化发育的不协调性,且此类细胞均高表达CD34.结论 急性淋巴细胞白血病伴异常表达髓系抗原的确诊必须依靠免疫学分型,同时该类原始细胞既具有典型B细胞的免疫标志又有髓系细胞的特有标志,具有髓系原始细胞及原始淋巴细胞的双重特点,是形态学误诊的最主要原因.     

应用流式细胞术评分诊断骨髓增生异常综合征

目的 采用流式细胞术（FCM）评分对骨髓增生异常综合征（MDS）与非MDS（主要为非克隆性血细胞减少患者）患者进行鉴别.方法 应用FCM四参数（CD34＋髓系原始细胞占所有有核细胞百分比、B系祖细胞占CD34＋细胞百分比、髓系原始细胞CD45表达水平、成熟粒细胞SSC峰通道值）对50例MDS患者和20例非MDS患者进行评分,每个参数异常时计1分,评分≥2分时诊断为MDS.结果与非MDS对照组比较,MDS患者CD34＋髓系原始细胞比例增高,B系祖细胞比例降低,成熟粒细胞SSC水平降低,差异有统计学意义（P＜0.05）;而CD34＋髓系原始细胞CD45表达水平差异无统计学意义（P=0.06）.MDS组FCM评分高于非MDS组,差异有统计学意义（P＜ 0.001）.45例患者用此方法正确诊断为MDS,敏感度为90％;1例患者为假阳性,特异度为95％.结论 采用FCM四参数评分可帮助诊断MDS,可行性好,简单方便.     

流式细胞免疫分型联合PCR检测抗原受体基因重排在淋巴增殖性疾病诊断与鉴别诊断中的应用

背景与目的:淋巴增殖性疾病是一组病理形态、免疫表型、临床特征高度异质性的疾病.本文旨在评价流式细胞免疫分型联合聚合酶链反应(PCR)检测抗原受体基因重排在淋巴增殖性疾病诊断中的应用价值.方法:分离79例拟诊淋巴增殖性疾病患者的新鲜病理标本(淋巴结、脾脏、胃活检组织和肝穿组织),流式细胞术分析细胞的免疫表型特征;同时采用BIOMED-2多重PCR分析组织的抗原受体基因重排状况及其克隆来源;并与病理诊断及免疫组织化学结果进行比对.结果:患者最终确诊为非霍奇金淋巴瘤(non-Hodgkin's lymphoma,NHL) 37例(46.8％),霍奇金淋巴瘤(Hodgkin's lymphoma,HL)3例(3.8％),淋巴组织良性病变34例(43.0％),恶性肿瘤淋巴结转移4例(5.1％).在19例B-NHL确诊患者中,流式免疫分型和抗原受体基因重排诊断符合率均为94.7％(18例);18例T-NHL确诊患者中,流式免疫分型和抗原受体基因重排诊断符合率分别为72.2％(13例)和50％(9例);在34例淋巴组织良性病变的患者中,流式细胞免疫分型虽未见单克隆幼稚细胞,但有7例(19.4％)抗原受体基因重排呈单克隆性;在3例HL确诊患者中,流式免疫分型未能检测出来,抗原受体基因重排均为多克隆性;4例恶性肿瘤转移患者中(3例转移浸润淋巴结,1例转移浸润肝脏),流式细胞免疫分型均检测到大量异常幼稚细胞,其中3例无特殊抗体标记,另外1例表达幼稚髓系标记;抗原受体基因重排均为多克隆性.结论:应用流式细胞免疫分型联合抗原受体基因重排分析,对于淋巴组织良、恶性增殖的鉴别和不同类型恶性淋巴瘤的诊断,具有很好的辅助价值.     

异常核型骨髓增生异常综合征64例预后分析

 【摘要】　目的　探讨骨髓增生异常综合征（MDS）患者染色体异常与预后的关系,对治疗效果进行分析。方法　回顾性分析122例MDS患者染色体核型,用吉姆萨显带法进行检测。难治性贫血（RA）、环形铁幼粒细胞难治性贫血（RAS）的治疗以诱导分化剂及刺激造血药物为主。原始细胞过多难治性贫血（RAEB）、转化型原始细胞过多难治性贫血（RAEB-t）、慢性粒-单核细胞白血病（CMML）的治疗以小剂量化疗和小剂量联合化疗方案为主。分析异常核型MDS患者疗效,以同期住院的正常核型MDS患者为对照。结果　检出异常核型MDS患者64例,治疗后完全缓解（CR）17例,CR率26.6 %。同期正常核型MDS患者58例,CR 30例,CR率51.7 %。正常和异常核型患者CR率差异有统计学意义（χ2＝8.13,P＝0.04）。复杂核型、-7、+8核型异常者易进展为急性白血病。结论　染色体核型分析在MDS的诊断与预后判断中有重要意义,不同的染色体核型改变进展为白血病的风险不同。     

骨髓增生异常综合征细胞形态学及细胞化学特征的研究

 目的　探讨骨髓增生异常综合征（MDS）的细胞形态学及细胞化学改变,为MDS提供形态学诊断依据。方法　对53例已确诊MDS患者的骨髓涂片进行瑞特-吉姆萨染色及铁染色,外周血中性粒细胞碱性磷酸酶（NAP）染色,分析骨髓细胞形态学特点。结果　53例MDS骨髓细胞粒、红、巨核三系均有不同程度的病态造血表现,分别有35、30、33例,骨髓各系列中出现病态造血频率较高的排列次序为粒系>巨核细胞系>红系,骨髓中细胞形态学病态造血改变类型的发生率比例较高的依次为单圆核巨核细胞29例（54.7 %）、假Pelger-Hu?觕t核28例 （52.8 %）、红系巨幼样变25例（47.2 %）等。46例（86.8 %）的MDS患者NAP染色积分及阳性率减低或为正常值下限。结论　多数MDS有两系以上的病态造血改变以及NAP染色积分及阳性率改变。     

世界卫生组织标准在骨髓增生异常综合征诊断分型中的应用

 【摘要】　目的　依据世界卫生组织（WHO）标准对249例骨髓增生异常综合征（MDS）进行诊断分型。方法　根据WHO（2008）标准对249例MDS行细胞形态学、细胞遗传学、免疫表型分析及骨髓病理活组织检查诊断。结果　所有MDS病例外周血成熟红细胞均见有巨大形与椭圆形改变;幼红、幼粒细胞、Pelger样核异常细胞与无颗粒中性粒细胞检出率随分型进展而增高;骨髓发育异常细胞特点及比例随MDS亚型进展而更明显、更高; 经动态随访观察, MDS随分型进展其转急性白血病（AL）率增高（P＜0.05）;148例患者行免疫表型分析,随分型进展髓系特异性抗原（CD33）表达逐渐增加;138例患者行染色体检查,其中异常核型表达53例（38.7 %）,主要见于20q-和+8,复杂异常核型16例（28 %）,其中5q- 2例;180例患者同时行骨髓活组织检查,19例具有形态异常,符合MDS诊断,42例伴有骨髓纤维化。结论　依据WHO标准多指标联合检测MDS有助于更准确的分型及诊断,进行长期追踪随访有助于判断预后。     

骨髓增生异常综合征患者重现性流式细胞术评分与修订版国际预后评分系统分层相关性

目的 研究骨髓增生异常综合征(MDS)患者预后分层和重现性流式细胞术评分系统(FCM-score)相关性.方法 采用FCM-score四参数[髓系前体细胞和有核细胞的比值(参数1)、B系前体细胞和CD34+细胞的比值(参数2)、淋巴细胞和髓系前体细胞CD45平均荧光强度的比值(参数3)、粒细胞和淋巴细胞SSC峰值的比值(参数4)]对54例MDS患者进行评分,用修订版国际预后评分系统(IPSS-R)对MDS患者进行预后评估,并对FCM-score和各个阶段(由IPSS-R评估得到)的MDS进行相关性分析.结果 FCM-score评分和IPSS-R分层之间存在正相关性(r=0.572,P<0.05),FCM-score四参数中参数1和参数4与IPSS-R分层之间存在正相关性(P<0.05).结论 FCM-score和IPSS-R分层存在相关性,可对MDS患者危险度进行初步评估.     

骨髓增生异常综合征和急性髓系白血病细胞遗传学特征研究

 【摘要】　目的　分析骨髓增生异常综合征（MDS）和急性粒-单核细胞白血病[急性髓系白血病（AML）-M4]、急性单核细胞白血病（AML-M5）患者的细胞遗传学特征。方法　采用骨髓细胞短期培养法,结合R显带技术对100例AML-M4、46例AML-M5与115例MDS患者进行细胞遗传学特征分析。结果　有26 %（26／100）AML-M4患者发生了克隆性细胞遗传学异常,常见的异常为+8、t（8;21）、-7;有26 %（12／46）AML-M5患者发生了克隆性细胞遗传学异常,常见的异常为+11;39 %（45／115）MDS患者发生了细胞遗传学异常,常见的异常为+8、亚二倍体、-7。结论　对MDS和AML患者进行细胞遗传学特征的分析对于其诊断具有重要的价值。     

骨髓增生异常综合征免疫表型的研究

目的：评价免疫表型测定在骨髓增生异常综合征(MDS)诊断及分型中的价值。方法：应用单克隆抗体方法对55例MDS患者进行免疫表型检测。结果：MDS患者髓系抗原表达明显增高,FAB亚型的抗原表达呈现规律性改变,随着RA向RAEB／RAEB-t转化,较早期的髓系抗原(CD33)逐渐增加,较成熟的CD15抗原和T-淋巴细胞抗原逐渐减少;同时在RAEB/RAEB—t阶段CD34^ 细胞数明显增高;较早期的骨髓细胞表面抗原(CD38、HLA-DR)在MDS表达明显增加。结论：免疫表型的检测对MDS更精确的诊断和分型有重要意义。     

Flow cytometric scoring system as a diagnostic and prognostic tool in myelodysplastic syndromes

The aim of this study is to validate the clinical utility of the flow cytometric scoring system (FCSS), quantifying phenotypic aberrancies in the myelomonocytic lineages, in the diagnosis and prognosis for conventionally treated myelodysplastic syndromes (MDS) patients. The bone marrow samples from 56 consecutive newly diagnosed MDS patients were characterized by the FCSS and compared with findings in 27 non-MDS cytopenic patients. The FCSS scores were significantly higher in patients with MDS than those in the non-MDS control. A flow score of 2 or more allowed for a specificity of 100% with 75% sensitivity in distinguishing these two groups. The FCSS scores correlated directly with validated prognostic systems including WHO classification, International Prognostic Scoring System (IPSS), WHO-adjusted prognostic scoring system (WPSS) and transfusion dependency. The median survival of conventionally treated MDS patients was directly related to FCSS group; severe: 6 months; moderate: 19 months and normal/mild: not reached. The multivariate analyses suggested the FCSS risk categories were an independent prognostic factor after adjustment for sex, age (above or below 70 years), IPSS or WPSS risk categories. These results confirm that quantifying aberrancies in the myelomonocytic lineage by FCSS is useful in MDS diagnosis and extends the prognostic utility for conventionally treated/untreated patients, especially among patients classified within the refractory cytopenia with multilineage dysplasia (RCMD) subgroup.     

Validation of a scoring system to establish the probability of myelodysplastic syndrome in patients with unexplained cytopenias or macrocytosis

Myelodysplastic syndromes (MDS) are clonal hematopoietic stem cell disorders primarily seen in the elderly that are characterized by ineffective hematopoiesis and a propensity to develop AML. Clinicians may be hesitant to refer older patients with unexplained cytopenias and/or macrocytosis for a bone marrow biopsy (BM), and consequently undiagnosed patients may be deprived access to effective treatments. Previously, we described factors which were independently predictive of a diagnosis of MDS at time of bone marrow: age ≥65, mean cell volume (MCV), red cell distribution width (RDW), and lactate dehydrogenase (LDH), and suggested a scoring system to calculate the post-test probability of MDS [1]. In this study we validate this scoring system in a cohort of 313 individuals who underwent bone marrow examinations for the investigation of unexplained cytopenias and or macrocytosis over a 3 year period at our institution (2006–2008). Thirty-two percent of all patients were diagnosed with MDS and 9% had suspected MDS. The post-test likelihood of a diagnosis of MDS increased from 12% when none of the four identified factors were present to 48% when 3 or more factors were present. This scoring system can be used to guide the diagnostic testing of patients presenting with unexplained cytopenias or macrocytosis.     

全基因组芯片检测在MDS诊断及预后中的应用

目的:探讨全基因组芯片（SNP-A）检测在骨髓增生异常综合征（MDS）患者诊断方面的特异性、敏感性及预后中的应用价值。方法:对2019年09月到2020年04月期间我院诊治的20例初诊MDS患者,依据修订的国际预后积分系统（IPSS-R）对患者进行危险度分组,应用SNP-A对MDS患者进行全基因组范围 DNA 拷贝数的变异（CNV）和单亲二倍体（UPD）的检测,总结特点并与常规染色体核型分析（CCA）结果进行比较。结果:20例 MDS 患者染色体核型分析的异常检出率为40%,与SNP-A相结合染色体异常的检出率提高至65%,二者比较差异有统计学意义（P＜0.05）;低危组和高危组患者核型异常的检出率分别为15%和25%,与 SNP-A相结合异常检出率分别提高至30%和35%（P＜0.05）;SNP-A检测出CNV和UPD高危组患者均明显高于低危组患者（P＜0.01）。结论:SNP-A与CCA的联合使用,大大提高了MDS患者染色体变异的检出率, 并且SNP-A能够对 MDS 患者提供更多更全面的遗传学信息,对于疾病的诊断和预后有重要价值。     

Primary myelodysplastic syndromes: analysis of prognostic factors in 235 patients and proposals for an improved scoring system.

In an attempt to identify prognostic factors for survival and leukemic transformation, 235 untreated patients with primary myelodysplastic syndromes (MDS) were analyzed in a single center retrospective study. To the well known FAB classification of MDS a supplementary group of patients with pure sideroblastic anemia (PSA) was added, characterized by the absence of dysplastic features of non-erythroid cells. Accordingly, the morphological subtypes were refractory anemia (RA), n = 55; PSA, n = 40; RA with ring sideroblasts (RARS), n = 33; RA with excess of blasts (RAEB), n = 53; RAEB in transformation (RAEB/T) n = 29; and chronic myelomonocytic leukemia (CMML), n = 25. Having screened 28 clinical, cytological, and laboratory parameters by univariate analysis, multiple regression analysis identified six variables with independent prognostic value: percentage of bone marrow blasts, serum LDH activity, PSA, hemoglobin concentration, age, and platelet count. If patients with PSA were excluded, the FAB classification no longer contributed independent prognostic information. Based on the results of this multivariate analysis, a simple scoring system was devised for predicting the survival of patients with MDS. A score of unity was allocated to each of the following parameters: bone marrow blasts greater than or equal to 5%, LDH greater than 200 U/I, hemoglobin less than or equal to 9 g/dl, and platelets less than or equal to 100 x 10(9)/I. As a function of their total score, patients were divided into three risk groups (group A, score 0; group B, score 1-2; group C, score 3-4), which differed significantly in both survival and rates of leukemic transformation. The cumulative survival 2 years after diagnosis was 91% in group A, 52% in group B, and 9% in group C (p less than 0.00005). The actuarial risk of transformation to acute myeloid leukemia at 2 years was 0, 19, and 54%, respectively (p less than 0.05). The inclusion of LDH enzyme levels qualified this scoring system for an accurate assessment of patients with CMML whose prognosis is viewed too favorably when rated by other scores. Furthermore, this score was able to identify those patients with RA and RARS who, without showing an excess of marrow blasts, have an unfavorable prognosis.     

Analysis of WHO-Based Prognostic Scoring System （WPSS） of Myelodysplastic Syndrome and Its Comparison with International Prognostic Scoring System （IPSS） in 100 Chinese Patients

Objective: The aims of this study were to assess the prognostic significance of WHO-based Prognostic Scoring System (WPSS) in myelodysplastic syndrome (MDS) from a single center institute and to compare WPSS with the international prognostic scoring system (IPSS).Methods: A total of 100 cases with de novo MDS were reviewed and their karyotypes were detected. All of them were followed up and classified according to IPSS and WPSS risk groups. SPSS 13.0 software was applied to deal with all the data. The statistical methods included Kaplan - Meier, Log-rank test and cox regression.Results: Multivariate cox regression analysis indicated that WHO Classification (P=0.0190), karyotype abnormalities categorized according to IPSS (P=0.0159) and red blood cell (RBC) transfusion (P=0.0009) were the three most important independent factors for predicting overall survival (OS) of MDS. WPSS and IPSS both had great capacity in predicting the OS of MDS at the time of diagnosis (P<0.0001). In time-dependent analysis, WPSS can predict the OS accurately in the following three years after diagnosis (P<0.0001), while IPSS failed to predict the OS 24 months after diagnosis (P=0.1094).Conclusion: Our single center results proved that WPSS is a dynamic prognostic system which can predict the OS of MDS patients at any time during the course of their disease. This time-dependent prognostic scoring system may replace the IPSS in the near future.     

骨髓增生异常综合征患者骨髓单个核细胞CD34、CD117的表达及其临床意义

 目的　探讨骨髓增生异常综合征（MDS）患者骨髓单个核细胞CD34、CD117的表达及临床意义。方法　采用直接免疫荧光标记法标记细胞表面分化抗原,流式细胞术测定,对37例MDS患者的骨髓单个核细胞CD34、CD117表达及临床意义进行分析。依据MDS的WHO分型方案、染色体核型以及国际预后积分系统（IPSS）将MDS患者划分为RA-RARS-RCMD 组、RAEBⅠ-RAEBⅡ组;染色体良好组、染色体不良组;中危Ⅰ组、中危Ⅱ组、高危组。结果　RA-RARS-RCMD组19例中11例CD34、CD117表达阳性,RAEBⅠ-RAEBⅡ组18例患者CD34、CD117表达均阳性;随着疾病恶性程度的增高,CD34、CD117表达明显升高;染色体良好组22例中14例患者CD34、CD117表达阳性,染色体不良组15例患者均表达CD34、CD117;随着异常克隆的出现,CD34、CD117表达升高;中危Ⅰ组17例中9例CD34、CD117表达阳性;中危Ⅱ组11例、高危组9例,所有患者CD34、CD117表达均阳性;随着预后积分的递增,CD34、CD117表达随之升高。结论　CD34、CD117检测有助于MDS 的分型、预后判断。     

流式细胞术用于良恶性胸腔积液鉴别诊断的研究--DNA倍体分析和突变型p53蛋白检测

目的研究流式细胞术DNA倍体和细胞周期分析、突变型p53检测用于良恶性胸腔积液鉴别诊断的临床可行性.方法 24例标本(12例恶性,12例良性)采用流式细胞术细胞内抗原检测的直接免疫荧光标记法检测胸腔积液细胞中表达突变型p53的阳性细胞百分率和p53表达的平均荧光强度,并且通过PI染色分析细胞倍型和周期分布.结果流式细胞术DNA倍体分析用于恶性胸腔积液诊断的敏感性为66.7%,特异性100%.倍体分析和细胞学检查联合应用能将诊断的敏感性提高到100%,特异性仍为100%.良性胸腔积液和恶性胸腔积液p53表达有显著性差异.以p53阳性细胞百分率＞90%为阳性标准,p53用于良恶性胸腔积液鉴别诊断的敏感性和特异性分别为83.3%和91.7%.联合p53阳性细胞百分率和DNA倍体分析对恶性胸腔积液诊断的敏感性能提高到91.7%,特异性91.7%.细胞周期SPF(S期时相百分比)在良恶性胸腔积液之间没有显著差异.结论流式细胞术DNA倍体分析和突变型p53检测可以作为临床良恶性胸腔积液鉴别诊断的辅助手段,结合细胞学检查更有意义.     

Multi-color CD34⁺ progenitor-focused flow cytometric assay in evaluation of myelodysplastic syndromes in patients with post cancer therapy cytopenia

Bone marrow assessment for myelodysplastic syndrome (MDS) in a patient who develops cytopenia(s) following cancer therapy is challenging. With recent advances in multi-color flow cytometry immunophenotypic analysis, a CD34(+) progenitor-focused 7-color assay was developed and tested in this clinical setting. This assay was first performed in 73 MDS patients and 53 non-MDS patients (developmental set). A number of immunophenotypic changes were differentially observed in these two groups. Based on the sensitivity, specificity and reproducibility, a core panel of markers was selected for final assessment that included increased total CD34(+) myeloblasts; decreased stage I hematogones; altered CD45/side scatter; altered expression of CD13, CD33, CD34, CD38, CD117, and CD123; aberrant expression of lymphoid or mature myelomonocytic antigens on CD34(+) myeloblasts; and several marked alterations in maturing myelomonocytic cells. The data were translated into a simplified scoring system which was then used in 120 patients with cytopenia(s) secondary to cancer therapy over a 2-year period (validation set). With a median follow-up of 11 months, this assay demonstrated 89% sensitivity, 94% specificity, and 92% accuracy in establishing or excluding a diagnosis of MDS.     

Evaluation of comorbidities at diagnosis predicts outcome in myelodysplastic syndrome patients

Recent data suggest that proper assessment of comorbidities is useful to predict the outcome of MDS patients receiving allogeneic stem cell transplantation. However, the results obtained in this highly selected subset of patients cannot be applied to the whole MDS population. We evaluated the impact of comorbidities in 418 consecutive MDS patients diagnosed at our institute from 1992 to 2005. All patients were classified according to WHO criteria and all received only conservative and supportive treatment. One or more comorbidities were detected in 390 patients (93%) at the time of diagnosis, with a higher incidence in older patients. Cardiac diseases were the most frequent comorbidities (30%) while diabetes and correlated adverse events were the second cause of comorbidity (20%). We applied 3 comorbidity prognostic scores (CCI, HCT-CI and a MDS-CI score proposed by Della Porta et al.). According to CCI score, 253 patients had a score 0, 111 patients had a score 1 and 54 patients had a score >2. According to HCT-CI, 209 patients had a score 0, 105 patients had a score 1 and 106 patients had a score >2. With MDS-CI score, 288 patients had a score 0 and 129 patients had a score >1. We found a significant correlation between survival and stratification according to CCI and MDS-CI scores (p = 0.01 and 0.02, respectively), but not according to HCT-CI score. The number of comorbidities as evaluated according to CCI was directly correlated to the development of RBC transfusion-dependency and was associated to a significantly higher risk of death not related to leukemic evolution (HR = 2.12, p ≤ 0.001). Conversely, higher risk of non-leukemic death did not correlate with higher transfusional requirement according to HCT-CI and MDS-CI scores (p = 0.3 and 0.43, respectively). As suggested by Della Porta et al., also in our experience the presence of cardiac, liver, renal, pulmonary diseases and solid tumours was found to independently affect the risk of death in a multivariable Cox regression analysis (p values from <0.01 to 0.004). In conclusion, assessment of comorbidities at diagnosis in MDS patients may improve the ability of therapeutic decisions.