血管平滑肌细胞体外摄取寡核苷酸探针的研究

目的:以99mTc标记针对c-myc和bcl-2 mRNA的寡核苷酸并检测其在血管平滑肌细胞(VSMC)中的摄取情况,初步探讨其在粥样硬化病变中的诊断价值。方法99mTc标记寡核苷酸后分别加入体外培养对数生长期及平台期猪冠状动脉VSMC,37℃条件下温育,检测不同时段摄取百分数。结果:增殖期或平台期VSMC对bcl-2探针的摄取均低于对c-myc探针的摄取。在c-myc探针组,增殖期细胞较平台期摄取高反义探针较正义探针摄取高。结论:增殖期VSMC对c-myc反义探针的摄取增高和(或)对bcl-2反义探针的摄取减少,有望成为粥样病变的早期诊断指标。     

Cytogenetics in patients with chronic myelogenous leukemia treated with bone marrow transplantation

Cytogenetic data are reported from 16 patients with Philadelphia chromosome (Ph) positive chronic myelogenous leukemia (CML) treated with bone marrow transplantation (BMT). The usefulness of cytogenetic investigations for the assessment of marrow engraftment is stressed. The significance of persistence or reappearance of Ph after BMT, possibly due to a defective leukemic clone eradication by the conditioning regimen, is also discussed. Generally, Ph-positive cells are damaged and disappear within the first year of BMT. Sometimes, however, the cells may repair the damage and proliferate again, resulting in disease relapse. Rarely, clinical and hematologic relapse does not follow Ph-positive clone expansion although leukemic cells represent more than 50% of marrow metaphases examined. Finally, the effect of interferon on Ph-positive clones after BMT and random chromosome changes, that appear transiently after BMT and are of uncertain significance, are discussed.     

Cytogenetic peculiarities in chronic myelogenous leukemia

Cytogenetic investigations were performed in 185 patients with chronic myelogenous leukemia (CML) at all stages of the disease; 166 patients were Ph positive-159 (95.8%) of these showing the standard Ph translocation, and 7 (4.2%) variant translocations-17 patients were Ph negative. In 2 patients the cytogenetic analysis was unsuccessful. Additional aberrations were found in 40 (24.1%) of the Ph-positive patients. Nine (52.9%) of the Ph-negative patients showed chromosome anomalies. Besides the well known nonrandom abnormalities (-7, +8, i(17q), +19, +Ph) we found a high frequency of clones with rare or not yet described structural rearrangements--in 14 cases (34.2%) of the Ph-positive patients and in 2 cases (20%) of the Ph-negative patients with other chromosome abnormalities. The clinical significance of these findings is discussed.     

慢性粒细胞白血病患者骨髓间质干细胞的生物学特征的研究

目的： 研究不同来源（正常人、慢性粒细胞性白血病）骨髓间质干细胞的生物学特征及分化能力。 方法: 分离获取19例慢性粒细胞性白血病（CML）患者的骨髓间质干细胞，同时获取8例正常人的骨髓间质干细胞作为对照。获取的间质干细胞采用低血清培养液培养。瑞士染色观察形态，FACS检测其免疫表型和细胞周期，通过油红O和von Kossa染色来证实向脂肪和骨的分化情况。RT-PCR检测CML特异性表达的BCR/ABL基因；对正常人、CML患者的骨髓间质干细胞进行染色体分析。 结果: 正常人和CML骨髓间质干细胞具有相似的细胞形态、生长特性和免疫表型，而且都可以向骨和脂肪分化。CML来源的MSCs不表达BCR/ABL融合基因，并且具有正常的核型。 结论: 从CML骨髓中可以分离和培养出具有间质干细胞特性的细胞群体，其具有正常的核型且不表达BCR/ABL基因。CML和正常人骨髓间质干细胞具有相似的生物学特性和多向分化能力。     

Antibody and Cell-Mediated Target Cell Lysis of Technetium-99m Labeled Erythrocytes

An assay for cell-mediated lysis of sheep erythrocytes has been developed using technetium-99m (^99mTc) as a radioisotopic label. Spleen cells obtained from sensitized BALB/c mice in combination with either non-immune or immune mouse anti-sheep RBC serum produced maximum release (100%) of ^99mTc. Non-immune spleen cells in combination with immune serum produced significantly greater release of ^99mTc compared to that observed with non-immune cells in the presence of non-immune serum. The kinetics of antibody-mediated complement dependent release of ^99mTc as a function of time and temperature paralleled the release of ⁵¹Cr and hemoglobin as an indicator of erythrocyte membrane injury. Because of its high specific activity, low spontaneous release, and ready availability in those institutions where diagnostic scanning is performed, ^99mTc appears to be particularly well suited for the assessment of both antibody and cell-mediated lysis of erythrocytes.     

A case of near-triploidy in chronic myelogenous leukemia

A 61-year-old woman with chronic myelogenous leukemia (CML) in accelerated phase had a near-triploid bone-marrow karyotype. This karyotype is an unusual finding in CML, as we review 12 previously published similar cases. These patients do not differ clinically from other patients with CML in blast crisis. The cytogenetic features of near-diploid and near-triploid CML are similar, except that relative loss of chromosomes is more common and that isochromosome 17q has not been reported in near-triploid CML.     

Preleukemic phase of chronic myelogenous leukemia: morphologic and immunohistochemical characterization of 7 cases

Patients with chronic myelogenous leukemia (CML) present typically with an elevated white blood cell count (WBC) and cytogenetic or molecular genetic evidence of t(9;22)/BCR-ABL1 fusion gene. Rarely, CML patients may present with a normal or mildly elevated WBC and are asymptomatic, and we describe 7 patients in this study. The WBC in these patients ranged from 3.6 to 14.3 K/mm³ with 50% to 73% granulocytes and 0% blasts. In all patients, t(9;22)(q34;q11.2) was detected by conventional cytogenetics, and BCR-ABL1 fusion was shown, supporting the diagnosis of preleukemic CML (pre-CML). We compared these patients with a group of 5 cases of CML in chronic phase (CML-CP) and 5 bone marrow specimens with a leukemoid reaction (n = 5). Reticulin, CD34, and CD61 immunostains were performed on all bone marrow biopsy specimens. Peripheral blood absolute basophilia (≥200/mm³) was noted in only 4 of 7 pre-CML cases, whereas it was present in all CML-CP cases and absent in leukemoid reaction cases. The mean ±SD of microvascular density of pre-CML cases (10.0 ± 4.3 vessels/200× field) was twice that of leukemoid reaction cases (5.0 ± 1.0) (P = .02; Student t test) but similar to that of CML-CP cases (12.5 ± 3.6). Microvessels in pre-CML, highlighted by CD34, were tortuous with abnormal branching, although to a lesser extent than those found in CML-CP. Microvessels in leukemoid reaction were generally straight. The percentage of small, hypolobated megakaryocytes, highlighted by CD61 in pre-CML, was 40%, 3 times that found in leukemoid reaction cases (13%) but less than that of CML-CP cases (86%). We conclude that pre-CML should be suspected in patients with a normal to mildly elevated WBC and absolute basophilia. Bone marrow examination can usually distinguish pre-CML from a leukemoid reaction based on the percentage of small, hypolobated megakaryocytes; microvascular density; and morphologic features.     

Karyotype at relapse following allogeneic bone marrow transplantation for chronic myelogenous leukemia.

Eighty-four patients underwent allogeneic or syngeneic bone marrow transplantation as therapy for chronic myelogenous leukemia (CML) during a 5-year period at The Johns Hopkins Oncology Center. We describe the karyotype at relapse in 19 patients who were Ph chromosome positive (Ph+) at diagnosis. Eighty-four percent of patients demonstrated clonal and/or nonclonal chromosome abnormalities in addition to the t(9;22)(q34;q11) at first detection of relapse or later during relapse. These abnormalities included: Ph plus additional clonal abnormalities (three patients), Ph plus nonclonal abnormalities (five patients), Ph plus additional clonal and nonclonal abnormalities (eight patients). Three patients had only the original Ph+ clone. The additional chromosome abnormalities were primarily structural, and entirely different from those most frequently observed during karyotypic evolution in conventionally treated CML. Chromosome 1 was most frequently involved, with 1q32 being the location of three clonal and two nonclonal abnormalities. Other sites included 6p21-22 (the site of two clonal abnormalities), 7p21-22, and 10q21 (the site of two clonal and one nonclonal abnormality each). Chromosomes 5 and 7q, regions of frequent involvement in acute nonlymphocytic leukemia that follows chemotherapy for other malignancies, were infrequently involved. The clinical significance of these additional abnormalities remains undetermined at this time.     

14q+ in Ph-positive chronic myelogenous leukemia

Two cases of chronic myelogenous leukemia with a Ph translocation and an additional chromosome change of the long arm of a chromosome #14 (14q+) are reported. The breakpoints on chromosome #14 were identified as 14q24 and 14q32, respectively. One of the patients did not show any evidence of blastic transformation; the other patient developed a myeloid blastic crisis when the abnormal 14q+ was seen in the bone marrow cells.     

Complex cytogenetic changes in Ph-negative chronic myelogenous leukemia

A Ph-negative chronic myelogenous leukemia (CML) with t(3;7)(q21;q32), t(4;9)(q21;q34), and del(8)(q22) is reported. This case is rather unusual for Ph-negative CML in being associated with complex chromosome changes. The patient was diagnosed as in the accelerated phase of CML. It will be important to study this malignant disorder in detail cytogenetically and molecularly in order to ascertain its nature and place among the myeloproliferative disorders.     

Breakpoint zone of bcr in chronic myelogenous leukemia does not correlate with disease phase or prognosis

The presence of a zone 4 rearrangement in the bcr of two chronic myelogenous leukemia (CML) cell lines and 25 CML patients was investigated by hybridization of HindIII-EcoRI double-digested DNA with the homologous HindIII-BglII breakpoint cluster region (bcr) probe. A rearrangement in zone 4 was found in the EM2 cell line and in 11 of the 25 patients. The presence of a zone 4 rearrangement was not associated with phase of disease, clinical evidence of acceleration, or pathologic evidence of acceleration. The difference between the actuarial duration of chronic phase for patients without (33 months) and with (22 months) a zone 4 rearrangement was not significant. A breakpoint in zone 4 of the bcr in patients with CML does not appear to be a determinant of prognosis.