Flow cytometric detection of immunoglobulin light chain in hematolymphoid immunophenotyping

During B-cell development and maturation,the antigen receptor,which is encoded by the immunoglobulin heavy-(IgH)and light-chain genes,rearrange to associate one of a number of variable,diverse,andjoining gene segments.Asingle mature Bcell expresses an IgHchain and either a kappa or lambda light     

克隆性基因重排检测在NHL诊断中的意义

目的探明PCR克隆性基因重排检测（基因诊断）在NHL诊断中的意义。方法应用病理组织学、免疫组化、基因诊断方法三结合方案分析178例与会诊31例NHL诊断的作用。结果分析结果显示：以病理组织学为主,辅以免疫组化确诊NHL114例。疑难、早期、微量标本共64例与会诊31例均作了基因诊断。总的阳性率为61．4％（58／95）。其中IgH阳性13例（22．4％）,TCRβ芝邱阳性41例（70．6％）,双克隆性4例（7％）。本室基因诊断送检率为35．8％（64／178）,总确诊率为93．7％（60／64）。结论本文分析证明基因诊断是NHL三结合诊断中确诊疑难病例及分型的最有效的方法。     

Bcl-2 gene rearrangements and apoptosis rates in patients with non-Hodgkin's lymphoma during chemotherapy

OBJECTIVES: Overexpression of the bcl-2 gene as a result of the t(14,18) translocation leads to neoplastic transformation by suppressing apoptosis. However, apoptotic cell death in response to chemotherapy has not been investigated. This study was planned with the aim to investigate the association between bcl-2 gene rearrangements and apoptotic changes during chemotherapy in patients with non-Hodgkin's lymphoma. DESIGN AND METHODS: Lymphocytes from 33 patients were collected before and during chemotherapy. Bcl-2 gene rearrangements were investigated by PCR. Apoptotic cell death was analyzed by enzyme immunoassay. RESULTS: In 24 cases, mbr gene rearrangements were detected. Apoptosis was successfully induced by chemotherapy in 48% of patients. Two characteristic, clearly distinguishable apoptotic response patterns with transient peaks either following the first or the third course were observed. It was found that apoptosis rates measured after the first course exactly reflect the final response. No correlation was found between bcl-2 gene rearrangements and apoptosis. CONCLUSIONS: Apoptotic cell death rates show transient changes during chemotherapy. Because the midterm response can be misleading, the apoptotic response should be evaluated following the first course of chemotherapy.     

Immunophenotyping by flow cytometry of fine needle aspirates in the diagnosis of lymphoproliferative disorders: A retrospective study.

In order to determine the value of flow cytometric (FCM) immunophenotyping of fine-needle aspirates (FNA) in the diagnosis and classification of lymphoproliferative diseases, 61 tissue samples were studied and compared with the cytologic/histological results. In vivo and ex vivo FNA biopsy yielded the material for FCM, which comprised an extensive number of lymphoid cell markers. In all but three cases sufficient cells were collected. Overall, malignancy was diagnosed in 33 cases from a total of 47 (70.2%), and in the remaining cases malignancy was not detected. Eleven cases were correctly diagnosed as reactive processes (11/11). There were no false positive cases of malignancy, as diagnosed by FCM-FNA. The best accuracy was achieved in the low-grade B-cell lymphomas and lymphoblastic lymphoma/leukemia. We conclude that in a significant number of cases, FCM-FNA permits the separation between lymphoid malignancies and reactive processes without false positive results. It was found to be particularly useful in the differential diagnosis of mantle-cell and small-lymphocytic lymphoma and in the identification of lymphoblastic lymphoma/leukemia.     

Flow cytometric findings in the floral variant of follicular lymphoma

The floral variant of follicular lymphoma (FL-F) is an unusual variant of follicular lymphoma. The malignant “follicles” have an unusual “floral” appearance due to the proliferation of surrounding and infiltrating small lymphocytes. This variant of malignant lymphoma has been supported by immunohistochemical immunophenotyping and gene-rearrangement studies; however, the flow cytometric findings have not previously been described. We report the flow cytometric findings in two cases of FL-F. The surrounding and infiltrating small lymphocytes represented mantle cells which, in one case, obscured monoclonality were CD10+, supporting a follicular center cell origin. J. Clin. Lab. Anal. 12:310–314, 1998. © 1998 Wiley-Liss, Inc.     

超声对腹部恶性淋巴瘤与转移性淋巴结肿大鉴别诊断的探讨

目的:通过回顾性分析发生在腹部的恶性淋巴瘤与转移性淋巴结肿大的声像图特征,探讨超声鉴别诊断要点。方法:超声检出131例腹部实性结节或肿块,对其中41例经病理证实的恶性淋巴瘤与90例经手术(原发癌灶)、穿刺活检病理证实的转移性淋巴结肿大进行对比,重点分析了8项声像图特征。结果:两组对比病灶的发生部位、大小、内部回声、边界是否清晰,均无明显差别;病灶有无包膜、融合现象、融合内部回声多呈蜂窝状,其内带状分隔境界清晰,发生部位是否多区域,均有明显的差别。结论:病灶多有包膜回声、融合现象、融合内部回声多呈蜂窝状,发生部位多区域,是腹部恶性淋巴瘤与转移性淋巴结肿大声像图主要鉴别点,能为临床选择治疗方案和预后提供依据,具有较大的临床价值。     

细针吸取细胞块及细针吸取细胞学联合应用对涎腺肿瘤的诊断价值

目的:探讨细针穿刺细胞块联合细针吸取细胞学对涎腺肿瘤的诊断价值。方法:有病理组织学对比的123例涎腺肿瘤患者,均行细针吸取细胞学和细胞块切片诊断,分析二者对涎腺肿瘤诊断的准确率。结果:二者联合诊断敏感性为95.1%,细针吸取细胞学诊断敏感性为91.0%。对涎腺肿瘤分型诊断中二者联合诊断准确率为92.6%,细针吸取细胞学诊断准确率为65.0%,比较差异有统计学意义(P<0.05)。其中对恶性肿瘤分型中,二者联合诊断准确率91.8%,细针吸取细胞学诊断准确率为56.7%,差异有统计学意义(P<0.05)。结论:细针吸取细胞块和细针吸取细胞学对涎腺肿瘤的诊断准确性均较高,二者联合应用对组织分型、分类诊断有很高的应用价值。     

Flow cytometric detection of platelet-reactive antibodies and application in platelet crossmatching

Background: Alloimmunization against HLA or platelet antigens can cause refractoriness to platelet transfusions in multiply transfused patients. Crossmatching of platelet concentrates is effective in overcoming this problem. Study Design and Methods: A flow cytometric assay was used for simultaneous detection of lymphocyte-reactive and platelet-reactive antibodies in a single sample using fluorescein isothiocyanate-labeled anti-IgG. This assay was compared with the monoclonal antibody-specific immobilization of platelet antigens (MAIPA) assay in selected sera containing HLA and platelet antibodies. In a further study, this assay was compared with lymphocytotoxicity test results from thrombocytopenic patients, for whom platelet concentrates were ordered. The results of both assays were then correlated with the 1-hour corrected count increment, with a corrected count increment greater then 7500 considered as an adequate transfusion response. Results: The results of the MAIPA and flow cytometric assay in detecting platelet-reactive antibodies correlated well (p<0.0001, r = 0.84). The sensitivity and specificity of the flow cytometric assay in detecting platelet-reactive antibodies were 94.7 and 96.3 percent, when the MAIPA assay was taken as a reference. In unselected sera from patients, the sensitivity and specificity of the flow cytometric assays were, respectively, 72.7 and 91.7 percent in detecting lymphocyte- reactive antibodies and 70.6 and 77.7 percent in detecting platelet- reactive antibodies, when the lymphocytotoxicity test was used as a reference. With regard to an adequate transfusion response, the sensitivities and efficiencies were 20.0 and 82.1 percent, 33.3 and 84.3 percent, and 70.0 and 88.6 percent for the lymphocytotoxicity test and the lymphocyte-reactive and platelet-reactive flow cytometric assays, respectively. Conclusion: Flow cytometric crossmatching appears to be an effective method of detecting platelet-reactive antibodies that may affect the success of platelet transfusions. This procedure is well-suited for routine conditions and can be performed within 2 hours.     

经纤维支气管镜针吸术对纵隔淋巴结肿大及支气管腔外肺肿瘤的诊断意义

目的 探讨经纤维支气管镜(纤支镜)行纵隔淋巴结及叶、段支气管腔外肺肿瘤针吸活检的诊断价值。方法103例在纤支镜检查过程中，对经CT检查的84例纵隔淋巴结肿大和19例叶、段支气管腔外新生物而管腔结构基本正常的患者行针吸术和刷检。结果针吸阳性诊断率为75．73％；刷检阳性诊断率为8．74％；针吸与刷检联合诊断阳性率为77．67％；24．27％的患者穿刺点有少许出血。结论经纤支镜针吸术是一种简便、安全、阳性诊断率高的检查技术。     

细针穿刺细胞学在乳腺肿块诊断中的临床应用

目的探讨细针穿刺细胞学检查(FANC)在乳腺肿块诊断中的应用价值及诊断体会。方法回顾分析554例乳腺肿块患者的FNAC,并对其FNAC进行做病理对照。结果 FNAC与病理组织学结果进行对比分析,总准确率96.0%,敏感度94.3%,特异度96.8%,假阴性率5.1%,潜在假阳性率3.2%。结论 FNAC可以部分替代乳腺癌术中冰冻切片,对部分良性肿块确定组织学类型。     

Fine needle aspiration biopsy and flow cytometry in the diagnosis of lymphoma.

Fine needle aspiration biopsy (FNAB) is emerging as a technique of potential value in the diagnosis of benign and malignant lesions in areas such as the breast and thyroid gland. Its place in distinguishing reactive from neoplastic lymphoid proliferations, when compared to the established practice of excision biopsy and histopathology, continues to undergo evaluation. Morphology alone discriminates poorly between atypical or lymphoproliferative disorders as seen in the presence of Epstein-Barr or human immunodeficiency virus. Furthermore the polymorphic populations of follicular lymphoma may mimic reactive changes. In addition previous classifications of these tumours using working formulation or Kiel classification relied heavily on architecture, which is a feature not reflected in cytology smears. The World Health Organisation approach includes clinical features, immunophenotyping and cytogenetic profiles to define neoplasms of immunohaematopoietic tissues. Flow cytometry on fine needle aspiration biopsy offers additional advantages in being rapid and objective in quantitatively as well as qualitatively documenting cell surface characteristics. All patients referred for this procedure to Tygerberg Academic Hospital with suspected nodal or extranodal lymphoma between January 2002 and December 2004 were analysed. In each case flow cytometry and cytomorphology were correlated with histopathology on tissue biopsy, bone marrow examination and clinical follow-up for confirmation of diagnosis. Results of the 124 cases were tabulated and statistically processed. Eighty-one met the inclusion criteria, thirteen (16.1%) were not malignant, two (2.5%) were falsely negative, two (2.5%) were equivocal needing histology and in the remaining sixty-four (79%) diagnosis was achieved. SUMMARY: Fine needle aspiration coupled with flow cytometry can reliably distinguish between nodal and extranodal neoplastic B-cell population. It is concluded that appropriate use, in a collaborative multidisciplinary setting, may eliminate the need for surgical procedures in many cases. CONCLUSION: These advances are not widely recognised and this is particularly true in South Africa. Accordingly, such an approach has been prospectively evaluated in the Western Cape showing that the combination of ready availability and diagnostic accuracy, after an initial learning curve, allow accurate characterisation of haematologic malignancies so that excision biopsy may be reserved for specific further studies to provide data not available from this less invasive procedure.     

GAG、MDA检测在非霍奇金淋巴瘤诊疗中的临床意义

目的探讨葡糖氨基葡聚糖(GAG)、丙二醛(MDA)在非霍奇金淋巴瘤(NHL)中的诊疗价值。方法利用酶联免疫吸附试验(ELISA)测定99例NHL患者和58名健康对照者的血清GAG、MDA水平,对其中20例滤泡型淋巴瘤(FL)患者化疗效果进行随访监测,并对测定结果用非参数秩和检验进行分析比较。结果 99例NHL患者血清中GAG和MDA水平显著高于正常对照组(P0.01),且在FL患者中升高尤为显著(P0.01)。20例FL患者随访测定GAG水平与疾病进展情况基本一致(符合率75%),而MDA的变化率与病情并不完全一致。结论 GAG和MDA在NHL诊断和疗效监测中有较好的临床应用价值。     

超声引导下细针穿刺活检在甲状腺结节的鉴别诊断中的价值

目的 评价超声引导下细针穿刺活检(FNAB)在甲状腺结节的鉴别诊断方面的临床价值.方法 回顾性分析64例经手术后病理证实的甲状腺结节.结果 64例病灶全部施行了超声波检查.有61例病灶施行了FNAB.在病灶的鉴别诊断中,灰阶超声波检查总的正确率为57.8%;而灰阶超声检查正确率与病灶直径的关系为10mm以下微小结节的正确率为14.3%,11～20mm结节的正确率为55.2%,21～30mm的正确率为75%,＞30mm的正确率为68.8%.以声像图为特征中的边缘不整,内部回声不均,内有钙化而考虑为恶性时,其正确率分别为65.9%、60.9%、62.5%.在病灶的鉴别诊断中,FNAB的正确率是91.7%.不同直径病灶的FNAB正确率分别为:＜10mm的为100%,11～20mm的为96.4%,21～30mm的为91.7%,＞30mm的为85.7%.结论 超声引导下FNAB在甲状腺结节的诊断中有较高的临床价值.     

超声引导下FNAC联合BRAFV600E基因突变检测在甲状腺结节诊断中的意义

目的探讨超声引导下细针穿刺细胞学(UG-FNAC)联合BRAFV600E基因突变检测在甲状腺结节术前诊断中的作用。方法回顾性分析300例甲状腺结节UG-FNAC检查结果及BRAFV600E基因突变检测结果,同时对其中120例手术切除病例的组织病理学进行分析。结果300例UG-FNAC结果按照Bethesda报告系统(2017版)进行分类,Ⅰ类:25例,占8.3%;Ⅱ类:53例,占17.7%;Ⅲ类:14例,占4.7%;Ⅳ类:7例,占2.3%;Ⅴ类:20例,占6.7%;Ⅵ类:181例,占60.3%。300例FNAC样本中,BRAF V600E检测173例(57.7%)为突变型,127例(42.3%)为野生型。120例手术切除病例的组织病理学结果,良性病变6例,占5%;恶性肿瘤114例,占95%;BRAFV600E在甲状腺乳头状癌(PTC)中的突变率为87.5%。结论UG-FNAC联合BRAFV600E基因突变检测是诊断甲状腺结节良、恶性准确有效的方法,具有重要的临床指导作用。     

精液生精细胞学检查与睾丸细针穿刺对无精症诊断的比较

目的探讨精液生精细胞检查对无精症诊断的应用价值及临床意义。方法分析81例无精子症患者精液生精细胞学检查结果，与睾丸穿刺活检结果对照分析。结果精液生精细胞学检查对梗阻性和非梗阻性无精症分型诊断的符合率分别为85％、95．1％。41例非梗阻性无精症中，生精阻滞阶段在精子细胞、精母细胞、精原细胞分别为14例、21例和6例。对同一患者精液细胞学检查与睾丸穿刺活检一致性分析中，生精细胞发育程度一致（P〈0．01）。结论精液生精细胞学检查能对非梗阻性无精症诊断、分型有很高的预示性价值，但对梗阻性无精子症诊断还需与其它指标进一步确诊。     

细针穿刺细胞学与抗酸染色检验在淋巴结核诊断中的比较

目的比较细针穿刺细胞学与抗酸染色在淋巴结核临床诊断中的应用价值。方法对100例淋巴结病变患者穿刺物标本应用细针穿刺细胞学和涂片抗酸染色法检测,对2种检测方法的结果进行比较分析。结果 100例淋巴结核细胞学分型不同时期,抗酸染色阳性例数不同,其中结核结节期44例中抗酸染色阳性2例;干酪样坏死期38例中抗酸染色阳性36例,其他各期抗酸染色阳性几乎为0。结论细针穿刺细胞学诊断淋巴结核不同时期抗酸染色阳性率不同,其中干酪样坏死期抗酸染色阳性率最高,细针穿刺细胞学和抗酸染色相结合在临床上有重要的应用价值。     

Value of preoperative fine needle puncture cytology in the diagnosis of highly malignant thyroid tumors

Highly malignant thyroid tumours are frequently diagnosed only at an advanced stage where radical resection is no longer feasible. By the wide application of fine needle aspiration biopsy as a diagnostic tool in nodular thyroid diseases occasionally early stages of these tumours can be diagnosed. Despite early detection in 23% of our patients with anaplastic carcinomas and sarcomas of the thyroid gland and subsequent radical resection an improvement in the long-term outcome was not achieved. In contrast, thyroidectomy and appropriate oncologial therapy may result in long-term survival of patients with early stages of thyroid lymphomas.     

细针穿刺细胞学与抗酸染色检验在淋巴结核诊断中的比较

目的比较细针穿刺细胞学与抗酸染色在淋巴结核临床诊断中的应用价值。方法对100例淋巴结病变患者穿刺物标本应用细针穿刺细胞学和涂片抗酸染色法检测,对2种检测方法的结果进行比较分析。结果 100例淋巴结核细胞学分型不同时期,抗酸染色阳性例数不同,其中结核结节期44例中抗酸染色阳性2例;干酪样坏死期38例中抗酸染色阳性36例,其他各期抗酸染色阳性几乎为0。结论细针穿刺细胞学诊断淋巴结核不同时期抗酸染色阳性率不同,其中干酪样坏死期抗酸染色阳性率最高,细针穿刺细胞学和抗酸染色相结合在临床上有重要的应用价值。     

经软腭超声引导细针穿刺诊断咽旁间隙肿瘤

目的探讨经软腭彩色多普勒超声(CDFI)诊断咽旁间隙肿瘤的应用价值.方法应用7.0 MHz腔内彩色多普勒超声探头对CT或MRI发现的40例(43个)咽旁间隙内可疑病变行软腭CDFI扫查.观察病变形态、大小、内部回声及血流情况,并行超声引导下细针穿刺(FNA)病理细胞学诊断.结果40例经CDFI扫查显示病变43个,FNA细胞学诊断良性病变6个,恶性病变32个,5个病变内未见癌细胞,其中2个病变CDFI检查高度怀疑为恶性病变,经鼻咽活检病理确诊为鼻咽癌.结论经软腭超声引导细针穿刺细胞学诊断是一种诊断鼻咽部肿瘤的新方法.当病变局限于咽旁间隙内时,FNA是对常规鼻咽活检一种有益的补充,可以提高鼻咽癌的早期诊断率.     

针吸标本细胞块在免疫组化染色方法的探讨

目的探讨针吸标本细胞块通过免疫组化方法进行进一步诊断的方法。方法将细胞块制作方法与免疫组化染色方法相结合。结果细胞块行免疫组化染色其效果与活检标本完全相同。结论细胞块行免疫组化染色方法可行,明显提高了针吸病理的诊断率,已成为细胞学检查非常实用的实验技术。