伴染色体三体异常的慢性中性粒细胞白血病1例

慢性中性粒细胞白血病（chronic neutrophilic leukemia，CNL）是一种少见的骨髓增殖性疾病，其主要特点表现为外周血和骨髓中中性粒细胞的过度增生，同时可伴有肝脾肿大，而细胞遗传学和分子生物学检查无Ph染色体及BCR／ABL融合基因。这种疾病迄今为止全世界报道的病例不超过150例，虽然伴有9号染色体三体异常的CNL国外有报道，但国内尚未见报道。我院近期诊断1例伴有9号染色体三体核型异常的CNL，现报告如下。     

女性A型血友病FⅧ基因突变分析

报道罕见的女性A型血友病1例，并对其凝血因子Ⅶ进行基因突变分析。患者，女，65岁，因为跌倒后右胸痛2d入院，院内查体提示胸壁皮下血肿，双下肢等长，髋屈曲及内旋受限。测定凝血指标提示APTT61．3s，正常血浆纠正后为41．3s，而PT、FIB、TT均正常。有既往出血史。FⅧ活性为2％，FⅨ活性为200％，vWF：Ag为120％，vWF：RCof100％，vWF：CBAl28％，FⅧ结合分析正常；髋关节X片提示;双侧髋臼发育不良，髋关节骨关节炎。临床诊断为血友病A型。提取该患者外周血DNA，根据NM_000132之凝血因子FⅦ基因序列设计合成了其第14外显子特异的引物，行聚合酶链反应扩增，并对扩增产物进行测序分析，测序结果与标准序列进行比较，发现该患者出现4111A→C杂合突变，使1314位氨基酸由苏氨酸变为脯氨酸，产生一错意突变，该突变未见其它文献报道．     

PCR-DGGE法检测DNA碱基突变及多态性的方法学评价

目的 了解聚合酶链反应-变性梯度凝胶电泳(polymerase chain reaction-denaturing gradient gel electrophoresis, PCR-DGGE)法检测DNA碱基突变和分辨单核苷酸多态性(single nucleotide polymorphism, SNP)的能力.方法 分别用DNA序列分析法及PCR-DGGE法检测白血病患者治疗前及缓解期的白细胞线粒体DNA D-loop区,以缓解期为对照,了解两种方法检测治疗前白细胞线粒体DNA D-loop区突变的能力并作比较.同时将野生型和突变型DNA以不同比例混合模拟DNA多态性存在,以了解PCR-DGGE检测SNP的灵敏度.结果 以DNA序列分析作为对照,PCR-DGGE法检测碱基突变的特异度达到100%,灵敏度为97.1%;PCR-DGGE可检出单碱基突变;PCR-DGGE可检测出约0.5%的多态性存在.结论 PCR-DGGE是一个检测DNA碱基突变和多态性存在的较好的方法.     

血管性血友病四种指标实验检测意义的比较

目的探讨血管性血友病4种检测实验诊断方法的临床应用价值。方法使用vWF抗原水平检测(vWFAg)、vWF胶原结合分析实验(vWFCBA)、瑞斯托霉素辅因子活性测定(vWFRcof)、瑞斯托霉素诱导的血小板聚集试验(RIPA)4种方法同时对正常献血员、vWD患者及其他出血性疾病患者进行检测,比较其检测方法的优越性。结果4种检测血管性血友病的实验其结果在vWD患者组与正常献血员组和其他出血性疾病组比较P<001,差异均有统计学意义;正常献血员组与其他出血性疾病组比较P>005,差异无统计学意义。1型vWD患者组中4种检测方法相关性比较显示,vWFCBA与vWF∶Ag相关性最好(r=09610),其次为vWF∶Rcof与RIPA(r=09164),而vWF∶Ag与RIPA相关性最差(r=08132)。vWF∶CBA的变异系数(39%)最小,vWF∶Ag(41%)次之,而vWF∶Rcof和RIPA较高(分别为155%和173%)。4种检测方法与诊断的总符合率分别为vWF∶Ag857%,vWF∶Rcof762%,RIPA809%,vWF∶CBA952%。结论vWF∶CBA操作简便、重复性较好,在常规vWD的诊断分型中可替代vWF∶Rcof和RIPA。     

淋巴结细针穿刺标本的流式细胞术分析

Objective Comparative evaluation of flow cytometric immunophenotyping in the diagnosis and differentiation of lymphadenopathy,lymphoma and reactive lymphoid hyperplasia. Methods Ninty-nine fine-needle aspiration specimens from patients with tentative clinical lymphoprofierative disorders were consecutively analyzed by both cytology and flow cytometry with histology results as the gold standard. The three color antibodies including CD3,CD3,CD4,CD5,CD10,CD19,CD20,CD23,CD45,K,λ,FMC7 and CD34 were used for cell composition evaluation and cells with abnormal phenotype. Lymphoma cases were classified according to new WHO classification and subtypes were categorized by immunophenotypic analysis. The results from flow cytometry and cytology were compared. Results By cytological study, 40 of 99 cases were diagnosed with lymphoma, 29 cases were diagnosed with metastatic carcinoma, and 30 cases were diagnosed with reactive lymphoid hyperplasia, necrosis or tuberculosis. Among them, 2 non-Hodgkin lymphoma(NHL) cases were misdiagnosed as reactive lymphoid hyperplasia by cytology. Biopsy was performed in 18 cases of NHL including 16 B-NHL and 2 T-NHL By flow cytometry study, 35 of 99 eases were diagnosed with lymphoma, including 4 cases of lymphoblast lymphoma, 1 case of T-cell lymphoma, and 30 eases of other B-NHL For those 30 cases of B-NHL, 28 cases showed monoclonal light chain expression, and k: λ orλ: k atios exceed 3: 1, and B-cell proportion was (73. 2±27. 2)%. Twenty-six cases could be sub-classified by immunophenotyped. Among 16 histologically confirmed B-NHL cases, only 2 cases diagnosed with follicular lymphoma showed discrepancy with flow cytometry results. In all cases diagnosed with reactive lymphoid hyperplasia and metastasis carcinoma , no abnormal lymphocytes can be found, and k: λ or k: λ ratios were less than 3: 1. Conclusions Fine-needle aspiration analysis with flow eytometrie immunophenotyping can be helpful in diagnosis and differential diagnosis as well as sub-classification of NHL     

MAC预处理方案在慢性粒细胞白血病异基因骨髓移植中的应用

我们用ＭＡＣ（马法兰、阿糖胞苷和环磷酰胺）方案和短程移植物抗宿主病（ＧＶＨＤ）预防，对２例慢性粒细胞白血病（ＣＭＬ）患者行异基因骨髓移植取得较好疗效。病例和方法１病例２例患者均为ＣＭＬ慢性期。供者分别为同胞姐、弟。供、受者间ＨＬＡ配型（Ａ、Ｂ、ＤＲ、...     

血细胞分析仪性能监控的建立及评价

目的:对血细胞分析仪进行性能监控,保证所有的检测结果准确可靠。方法:参照CAP(CollegeofAmericanPathologists)要求和卫生部《临床实验室管理办法》制定出包括仪器校准、室内质控、不同仪器和不同检测模式间新鲜全血比对、相关性比较、携带污染、精密度、卫生部临床检验中心室间质评、美国CAP的能力比对和线性检测等9项性能指标;对每项监控指标均有明确规定,需要的材料、每年完成的最少次数、操作步骤、计算及失败后的纠正措施等,具有可操作性和规范化。结果:CAP及卫生部临检中心回报结果均为100%。结论:建立血细胞分析仪性能监控在保证不同仪器及同一仪器不同检测模式,检测结果的一致和准确十分重要;CAP及卫生部临检中心回报结果取得的优秀成绩证明检测结果的准确可靠。     

万珂为主化疗方案治疗多发性骨髓瘤的临床研究

目的 研究蛋白酶体抑制剂硼替佐米(万珂,Velcade,V)为主的化疗方案治疗多发性骨髓瘤(MM)的疗效和不良反应. 方法 MM患者接受万珂为主的联合方案进行化疗.疗效评定按照EBMT/ABMT标准.结果 可评估疗效者12例,初治8例,复发难治4例.治疗总体反应率100%(12/12),其中,完全缓解(CR)5例(41.7%),接近完全缓解(nCR)2例(16.7%),部分缓解(PR)5例(41.7%).初治患者达到最佳反应所需疗程中位数为2个(2～3个),复发难治患者达到最佳反应所需疗程中位数为4个(3～5个).中位随访11月(4～27月),所有患者均存活.不良反应多为1～2级,包括周围神经病变(41.7%)、血小板减少(33.3%)、恶心呕吐(33.3%)及呼吸道感染(16.7%),经对症处理后好转.结论 万珂为主的化疗方案治疗初治和复发难治MM病例,起效较快,反应率和完全缓解率较高,不良反应轻,患者耐受好,值得推广.     

青蒿琥酯对人淋巴细胞白血病/淋巴瘤细胞抑制作用及机理的研究

目的 观察青蒿琥酯(ART)对白血病/淋巴瘤细胞株Raji、Jurkat和急性淋巴细胞白血病(ALL)原代细胞的增殖抑制作用,以及ART与长春新碱(VCR)、阿糖胞苷(Ara-C)的细胞毒协同效应,并探讨其作用机制.方法 MTT法观察ART对Raji、Jurkat、ALL原代细胞的增殖抑制效应及ART与VCR、Ara-C的协同效应.Wright-Giemsa染色光镜下及透射电镜观察细胞凋亡的形态变化,Rhodamine-123检测线粒体跨膜电位(MMP)变化,比色法检测细胞内caspase-3浓度变化.结果 ART在体外能显著抑制Raji、Jurket细胞的增殖,对ALL原代细胞亦具有增殖抑制作用.低浓度ART与VCR、Ara-C联合,能增加VCR、Ara-C的细胞毒作用.ART作用后B/T淋巴细胞白血病/淋巴瘤细胞光镜及电镜均表现出凋亡形态学改变,线粒体跨膜电位下降,细胞内caspase-3的表达增加,呈浓度依赖性.与对照组相比,差异具有统计学意义(P<0.05).结论 ART对淋巴细胞白血病/淋巴瘤细胞具有抑制作用,且与VCR、Ara-C联用具有协同效应,其机制与诱导肿瘤细胞凋亡有关.ART有望开发成治疗ALL/淋巴瘤的新药.     

淋巴结细针穿刺标本的流式细胞术分析

Objective Comparative evaluation of flow cytometric immunophenotyping in the diagnosis and differentiation of lymphadenopathy,lymphoma and reactive lymphoid hyperplasia. Methods Ninty-nine fine-needle aspiration specimens from patients with tentative clinical lymphoprofierative disorders were consecutively analyzed by both cytology and flow cytometry with histology results as the gold standard. The three color antibodies including CD3,CD3,CD4,CD5,CD10,CD19,CD20,CD23,CD45,K,λ,FMC7 and CD34 were used for cell composition evaluation and cells with abnormal phenotype. Lymphoma cases were classified according to new WHO classification and subtypes were categorized by immunophenotypic analysis. The results from flow cytometry and cytology were compared. Results By cytological study, 40 of 99 cases were diagnosed with lymphoma, 29 cases were diagnosed with metastatic carcinoma, and 30 cases were diagnosed with reactive lymphoid hyperplasia, necrosis or tuberculosis. Among them, 2 non-Hodgkin lymphoma(NHL) cases were misdiagnosed as reactive lymphoid hyperplasia by cytology. Biopsy was performed in 18 cases of NHL including 16 B-NHL and 2 T-NHL By flow cytometry study, 35 of 99 eases were diagnosed with lymphoma, including 4 cases of lymphoblast lymphoma, 1 case of T-cell lymphoma, and 30 eases of other B-NHL For those 30 cases of B-NHL, 28 cases showed monoclonal light chain expression, and k: λ orλ: k atios exceed 3: 1, and B-cell proportion was (73. 2±27. 2)%. Twenty-six cases could be sub-classified by immunophenotyped. Among 16 histologically confirmed B-NHL cases, only 2 cases diagnosed with follicular lymphoma showed discrepancy with flow cytometry results. In all cases diagnosed with reactive lymphoid hyperplasia and metastasis carcinoma , no abnormal lymphocytes can be found, and k: λ or k: λ ratios were less than 3: 1. Conclusions Fine-needle aspiration analysis with flow eytometrie immunophenotyping can be helpful in diagnosis and differential diagnosis as well as sub-classification of NHL