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[Microcytofluorometrical measurement of DNA content in immunologically identified micromegakaryocytes of human bone marrow]. [免疫鉴定的人骨髓微巨核细胞DNA含量的微细胞荧光测定]。
Y Kobayashi, M Ozawa, N Maruo, M Kondo

We have developed a method to microfluorometrically determine the amount of DNA in immunologically identified micromegakaryocytes on bone marrow smears. Bone marrow smears were fixed with acetone-formalin buffer and immunostained with a monoclonal anti-GPIIb/IIIa antibody, followed by FITC conjugated anti-mouse IgG. After the smears were re-fixed with methanol, DAPI (4', 6-diamidino-2-phenylindole) staining was performed. Using an automatic Digital-Microfluorometer (Olympus MMSP-FR-II), megakaryocytes on the smears were identifies by the GPIIb/IIIa immunofluorescence and, after changing the barrier filters, their nuclear DNA content was measured by the intensity of DAPI fluorescence, which is proportional to the amount of DNA. Using this method, we found that the DNA histogram of the megakaryocytes from a patient with myelodysplastic syndrome showed a shift to small ploidy compared with normal controls. This method may be valuable in the measurement of the megakaryocyte DNA content.

我们已经开发了一种方法,以微荧光法确定骨髓涂片上免疫鉴定的微巨核细胞的DNA量。骨髓涂片用丙酮-福尔马林缓冲液固定,用抗gpiib /IIIa单克隆抗体进行免疫染色,然后用FITC偶联抗小鼠IgG进行免疫染色。涂片用甲醇重新固定后,进行DAPI(4′,6-二氨基-2-苯基吲哚)染色。使用自动数字微荧光仪(Olympus MMSP-FR-II),对涂片上的巨核细胞进行GPIIb/IIIa免疫荧光鉴定,更换屏障过滤器后,通过DAPI荧光强度测量其核DNA含量,DAPI荧光强度与DNA量成正比。使用这种方法,我们发现来自骨髓增生异常综合征患者的巨核细胞的DNA直方图显示与正常对照相比,向小倍体转移。该方法可用于巨核细胞DNA含量的测定。
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引用次数: 0
Plasma levels of von Willebrand factor and plasminogen activator in patients with arterial thromboembolism--with special reference to their correlation to the increased catabolism of both fibrinogen and platelets. 动脉血栓栓塞患者血浆血管性血友病因子和纤溶酶原激活剂水平——特别参考它们与纤维蛋白原和血小板分解代谢增加的相关性
M Matsumoto, T Uchiyama, N Kobayashi, T Maekawa

Plasma levels of von Willebrand factor(vWF) and plasminogen activator(PA) in the plasma of venous blood before and after 5 minutes' venous occlusion were studied in 104 patients with various types of arterial thromboembolic disease and 30 age-matched healthy subjects. Levels of beta-thromboglobulin(beta-TG) were determined in plasma prior to the venous occlusion. In 29 patients and 7 control subjects, turnover of intravenously injected 125I-labeled fibrinogen was studied. Mean plasma levels of von Willebrand factor antigen(vWF: Ag) and ristocetin cofactor activity(vWF: RCo) were significantly higher in patients than in controls both before and after the venous occlusion. Mean plasma PA activity was significantly lower in patients than in controls both before and after the venous occlusion, but mean plasma PA antigen before the venous occlusion was significantly higher in patients than in controls. Plasma clearance of i.v. injected 125I-labeled fibrinogen was significantly accelerated, and the catabolic flux(j3x) of fibrinogen calculated according to a two-compartment model was significantly higher in patients than in controls. Significant relationships were observed between T1/2 of 125I-labeled fibrinogen and the following: plasma levels of vWF: Ag both before and after the venous occlusion, PA activities after the occlusion, PA antigen before the occlusion, and the net decrease in PA activities and the net increase in PA antigen as a result of the occlusion. Significant relationships were also observed between j3x of fibrinogen and the following: plasma levels of vWF: Ag both before and after the venous occlusion, vWF: RCo after the occlusion, PA activities after the occlusion, PA antigen before the occlusion, and the net decrease in PA activities resulting from the occlusion. Plasma levels of beta-TG, which were significantly higher in patients than in controls, were not correlated to plasma levels of vWF and PA or to parameters of fibrinogen turnover. These results suggest that the change in endothelial cell function is responsible for the abnormal plasma levels of both vWF and PA and for the acceleration of fibrinogen metabolism in patients with thromboembolic disease.

本文研究了104例不同类型动脉血栓栓塞性疾病患者和30例年龄匹配的健康人在静脉闭塞5分钟前后静脉血血浆中血管性血友病因子(vWF)和纤溶酶原激活剂(PA)的水平。静脉闭塞前测定血浆中β -血栓球蛋白(β - tg)水平。在29例患者和7例对照者中,研究了静脉注射125i标记纤维蛋白原的周转。静脉闭塞前后,患者平均血浆血管性血友病因子抗原(vWF: Ag)水平和利斯托司汀辅助因子活性(vWF: RCo)均显著高于对照组。静脉阻断前后患者血浆平均PA活性均明显低于对照组,但静脉阻断前患者血浆平均PA抗原明显高于对照组。静脉注射125i标记纤维蛋白原的血浆清除率明显加快,根据双室模型计算的纤维蛋白原分解代谢通量(j3x)在患者体内明显高于对照组。125i标记纤维蛋白原T1/2与静脉阻断前后血浆vWF: Ag水平、阻断后PA活性、阻断前PA抗原、阻断后PA活性净降低和PA抗原净升高之间存在显著相关性。纤维蛋白原j3x与静脉阻断前后血浆vWF: Ag水平、阻断后血浆vWF: RCo水平、阻断后血浆PA活性、阻断前血浆PA抗原水平、阻断后血浆PA活性净下降呈显著相关。血浆β - tg水平在患者中明显高于对照组,与血浆vWF和PA水平或纤维蛋白原转换参数无关。这些结果表明,内皮细胞功能的改变是导致血栓栓塞性疾病患者血浆vWF和PA水平异常以及纤维蛋白原代谢加速的原因。
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引用次数: 0
Fatal graft-versus-host disease following transfusion during open heart surgery. 心脏直视手术输血后致死性移植物抗宿主病。
T Aso, Y Asano, M Harada, J Kudo, K Fujimoto, T Okamura, Y Tsuda, Y Niho

We report fatal transfusion-associated graft-versus-host disease (GVHD) in a patient who was not severely immunosuppressed. A 58-year-old man received 800 ml of fresh whole blood from his son and an unrelated volunteer donor during open heart surgery. On the 10th day after the operation, he suddenly had a high fever, followed by generalized skin rash and liver dysfunction. Pancytopenia due to bone marrow aplasia developed a week later. A skin biopsy revealed a cutaneous lesion highly compatible with acute GVHD. The patient did not respond to high-dose methylprednisolone therapy, and died of multiple organ failure on the 18th day after the operation.

我们报告了一名没有严重免疫抑制的患者的致命输血相关移植物抗宿主病(GVHD)。一名58岁的男子在心脏直视手术中接受了800毫升新鲜全血,来自他的儿子和一名无关的志愿献血者。术后第10天,患者突然出现高热,继发全身皮疹及肝功能障碍。一周后出现骨髓发育不全引起的全血细胞减少症。皮肤活检显示皮肤病变与急性GVHD高度一致。患者对大剂量甲基强的松龙治疗无反应,于术后第18天死于多器官功能衰竭。
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引用次数: 0
Clinical and cytogenetic features in six patients with chronic myelogenous leukemia and a complex Philadelphia translocation. 6例慢性骨髓性白血病伴复杂费城易位的临床和细胞遗传学特征。
S Misawa, K Nishida, M Taniwaki, H Nishigaki, T Takino, S Nakanishi, C Shimazaki, M Nakagawa, J Inazawa, T Abe

Six patients with chronic myelogenous leukemia and a complex Philadelphia (Ph1) translocation are described. The complex Ph1 translocations were a three-way translocation in five patients and a five-way translocation in one. Additional chromosomal aberrations were detected in four of five patients when the blastic crisis supervened. The median survival time was 42 months. The remaining patient died of acute myocardial infarction 23.5 months after the diagnosis of CML was made. There seems no difference between these six patients and those with the standard Ph1 with respect to clinical, hematologic and cytogenetic findings.

6例慢性骨髓性白血病和复杂的费城(Ph1)易位的描述。复杂的Ph1易位在5例患者中为三向易位,在1例患者中为五向易位。当胚性危机发生时,在5例患者中有4例检测到额外的染色体畸变。中位生存时间为42个月。其余患者在诊断为CML后23.5个月死于急性心肌梗死。在临床、血液学和细胞遗传学方面,这6名患者与标准Ph1患者之间似乎没有差异。
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引用次数: 0
Systemic lupus erythematosus in a patient with refractory anemia. 难治性贫血患者的系统性红斑狼疮。
S Nakayama, F Nakamura, H Yabe, K Nagai

A 69-year-old man was diagnosed as having refractory anemia (RA), accompanied by pancytopenia of two years' duration, myelodysplasia in all three cell lines, abnormal karyotype of 46, XY, 20q--in bone marrow cells, and positive antinuclear and DNA antibody tests. One year after his first visit, he developed arthritis and maculopapular erythematous rashes. The histologic features of the skin lesions were similar to those of in discoid lupus erythematosus. He was diagnosed as having systemic lupus erythematosus supervening refractory anemia.

一位69岁的男性被诊断为难治性贫血(RA),伴有持续两年的全血细胞减少症,所有三种细胞系骨髓增生异常,骨髓细胞核型异常46、XY、20q-,抗核和DNA抗体试验阳性。第一次就诊一年后,他出现了关节炎和黄斑丘疹。皮肤病变的组织学特征与盘状红斑狼疮相似。他被诊断为系统性红斑狼疮并发难治性贫血。
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引用次数: 0
Effects of recombinant human G-CSF and GM-CSF on primary human leukemic cells. 重组人G-CSF和GM-CSF对人白血病原代细胞的影响。
K Itoh, M Bessho, K Hirashima

The effects of recombinant human granulocyte colony-stimulating factor (rhG-CSF) and recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) on primary human leukemic cells were studied. Phagocyte-depleted mononuclear cells containing more than 88% blasts were obtained from peripheral blood of 11 AML and 2 ALL patients and from bone marrow aspirates from 2 ALL patients. The leukemic cells were incubated with these CSF in suspension cultures or in methylcellulose cultures. In suspension cultures, the spontaneous proliferation was observed in 1 M4 patient. RhG-CSF stimulated the leukemic cell proliferation in 5 AML, cases and rhGM-CSF that in 4 AML cases. In methylcellulose cultures, spontaneous colony formation occurred in 3 M4 patients. RhG-CSF and rhGM-CSF stimulated the leukemic colony formation in 8 AML cases. The CSFs had an additive effect in both cultures. Neither CSF induced O2- production or phagocytic activity. From these results, we concluded that both CSFs stimulated the proliferation of leukemic cells without inducing differentiation.

研究了重组人粒细胞集落刺激因子(rhG-CSF)和重组人粒细胞-巨噬细胞集落刺激因子(rhGM-CSF)对人白血病原代细胞的作用。从11例AML患者和2例ALL患者的外周血和2例ALL患者的骨髓抽吸液中获得了含有88%以上原细胞的吞噬细胞耗尽的单核细胞。白血病细胞与这些脑脊液在悬浮培养或甲基纤维素培养中孵育。在悬浮培养中,1例M4患者观察到自发增殖。RhG-CSF在5例AML中刺激白血病细胞增殖,在4例AML中刺激白血病细胞增殖。在甲基纤维素培养中,3例M4患者发生了自发的菌落形成。RhG-CSF和rhGM-CSF刺激8例AML患者白血病集落形成。csf在两种培养中均具有加性效应。脑脊液均未诱导氧生成或吞噬活性。从这些结果,我们得出结论,这两种csf刺激白血病细胞的增殖而不诱导分化。
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引用次数: 0
Effect of vitamin K deficiency on urinary gamma-carboxyglutamic acid excretion in rats. 维生素K缺乏对大鼠尿γ -羧谷氨酸排泄的影响。
M Yamano, Y Yamanaka, K Yasunaga, K Uchida

Since gamma-carboxyglutamic acid (Gla) in Gla-containing proteins is stoichiometrically excreted into urine as free Gla, urinary Gla excretion is believed to reflect the rate of synthesis and degradation of vitamin K-dependent proteins and the utilization of vitamin K in body. We studied the daily changes in urinary Gla excretion and plasma vitamin K-dependent clotting factor levels in rats fed vitamin K-deficient diets followed by subcutaneous injection of vitamin K1 or after the oral administration of Warfarin. Urinary Gla excretion in normal rats fed a standard diet that contained about 500 ng of vitamin K1 per gram of diet was 2.35 +/- 0.25 mumoles/day, but the level in rats fed a markedly vitamin K-deficient diet (less than 5 ng/g) decreased to 1.40 +/- 0.14 mumoles/day. When rats were fed a moderately vitamin K-deficient diet (20-50 ng/g), plasma vitamin K-dependent clotting factor levels decreased significantly, but urinary Gla excretion did not decrease. Warfarin, a vitamin K antagonist, caused a significant decrease in urinary Gla excretion and plasma clotting factor levels. When vitamin K, (200 micrograms/kg) was injected subcutaneously in rats fed a markedly vitamin K-deficient diet, the plasma vitamin K-dependent clotting factor levels recovered quickly to normal, but urinary Gla excretion showed only a partial recovery to 1.74 +/- 0.15 mumoles/day. These results indicate that urinary Gla excretion decreases in vitamin K deficiency, but changes in urinary Gla excretion do not reflect vitamin K deficiency in rats as sensitively as changes in the prothrombin time and plasma K-dependent clotting factor levels.

由于含Gla蛋白中的γ -羧谷氨酸(Gla)以游离Gla的形式排泄到尿中,因此尿中Gla的排泄量被认为反映了维生素K依赖性蛋白的合成和降解速度以及体内维生素K的利用情况。我们研究了在维生素k缺乏饮食后皮下注射维生素K1或口服华法林后大鼠尿Gla排泄和血浆维生素k依赖性凝血因子水平的每日变化。正常大鼠每克饮食中含有500纳克维生素K1的标准饮食,其尿玻璃排泄量为2.35 +/- 0.25 μ摩尔/天,而明显缺乏维生素k的大鼠(低于5纳克/克)的尿玻璃排泄量降至1.40 +/- 0.14 μ摩尔/天。当给大鼠喂食中度维生素k缺乏(20-50 ng/g)时,血浆维生素k依赖性凝血因子水平显著降低,但尿玻璃排泄量未减少。华法林,一种维生素K拮抗剂,导致尿玻璃排泄和血浆凝血因子水平显著下降。在明显缺乏维生素K的大鼠中皮下注射维生素K(200微克/千克),血浆维生素K依赖性凝血因子水平迅速恢复正常,但尿Gla排泄仅部分恢复到1.74 +/- 0.15 μ mol /d。这些结果表明,尿Gla排泄在维生素K缺乏时减少,但尿Gla排泄的变化并不像凝血酶原时间和血浆K依赖性凝血因子水平的变化那样敏感地反映维生素K缺乏。
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引用次数: 0
Aplastic anemia: lack of increase of in vitro colony formation after T cell depletion with monoclonal antibodies and complement. 再生障碍性贫血:单克隆抗体和补体T细胞耗竭后体外集落形成缺乏增加。
S Kojima, K Matsuyama, K Miyamura, Y Kodera

To detect suppressor T cells to hematopoietic stem cells, growth of granulocyte-macrophage colony-forming cells (CFU-GM) and burst-forming unit (BFU-E) was compared before and after treatment of bone marrow cells with anti-T monoclonal antibodies and complement in 29 patients with aplastic anemia. The anti-T monoclonal antibodies used were 35.1 (CD2), Tp120 (CD6) and ATL27 (not clustered). Treatment of normal bone marrow with anti-T monoclonal antibodies and complement resulted in complete (greater than 99%) lysis of T cells with negligible effects on colony growth. Preincubation of marrow samples with monoclonal antibodies and complement did not enhance CFU-GM or BFU-E colony growth in patients with aplastic anemia. Using this assay, there was no evidence of T cell-mediated inhibition of colony proliferation in any of 29 patients.

采用抗T单克隆抗体和补体治疗29例再生障碍性贫血患者,比较骨髓细胞治疗前后粒细胞-巨噬细胞集落形成细胞(CFU-GM)和突发性形成细胞(BFU-E)的生长情况,检测造血干细胞的抑制性T细胞。使用的抗t单克隆抗体为35.1 (CD2)、Tp120 (CD6)和ATL27(未聚集)。用抗T单克隆抗体和补体治疗正常骨髓导致T细胞完全(大于99%)溶解,对菌落生长的影响可以忽略不计。骨髓样品与单克隆抗体和补体预孵育不能促进再生障碍性贫血患者的CFU-GM或BFU-E集落生长。在29例患者中,没有T细胞介导的集落增殖抑制的证据。
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引用次数: 0
Glanzmann's thrombasthenia associated with a transient deficiency of factor XIII. 格兰兹曼凝血症与暂时性因子XIII缺乏有关。
Y Mizuno, T Hara, T Otsuki, H Tsuda, T Kajiwara, K Nibu, N Yoshida, S Miyazaki, K Ueda

A three-year-old girl suffering from ecchymoses developed severe epistaxis. The diagnosis of thrombasthenia was made on the basis of platelet aggregation studies, flow cytometric analysis with monoclonal antibodies and gel electrophoretic analysis. In addition, coagulation studies at the time of epistaxis repeatedly showed a transient deficiency of factor XIII activity and antigen.

一名患有淤血的三岁女孩发展为严重的鼻出血。血栓减少的诊断是基于血小板聚集研究、单克隆抗体流式细胞分析和凝胶电泳分析。此外,凝血研究在出血时反复显示暂时性缺乏因子XIII活性和抗原。
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引用次数: 0
1 alpha, 25-dihydroxyvitamin D3 enhances the up-regulation of interleukin-2 receptor (p55) by interleukin-2. 1 α, 25-二羟基维生素D3增强白介素-2对白细胞介素-2受体(p55)的上调。
S Tamori, T Uchiyama, H Uchino

The effect of 1 alpha, 25-dihydroxyvitamin D3 (1 alpha, 25(OH)2D3) on the expression of interleukin-2 (IL-2) receptor in activated T lymphocytes was examined. 1 alpha, 25(OH)2D3 enhanced the expression of IL-2 receptor (p55, Tac peptide) in phytohemagglutinin (PHA)-stimulated (3 days) human peripheral blood mononuclear cells (PBM) only in the presence of IL-2 without affecting the proliferation of the cells. This enhancement was dependent on the concentration of both IL-2 (0-1 U/ml) and 1 alpha, 25(OH)2D3(0-10(-7)M). The addition of interleukin-1 (IL-1, 0-100 U/ml), did not enhance the expression of IL-2 receptor in these cells in the presence of IL-2. Moreover, 1 alpha, 25(OH)2D3 had the same effect on two cell lines, Kit225 (an IL-2 dependent cell line established from a patient with T cell chronic lymphocytic leukemia) and YT (an IL-2 independent natural killer (NK)-like cell line from a patient with acute lymphocytic leukemia). Thus, 1 alpha, 25(OH)2D3 enhances the up-regulation of IL-2 receptor (p55) by IL-2 not only in activated T cells but also in the NK-like cell line.

研究了1 α, 25-二羟基维生素D3 (1 α, 25(OH)2D3)对活化T淋巴细胞白细胞介素-2 (IL-2)受体表达的影响。1 α, 25(OH)2D3仅在IL-2存在的情况下增强植物血凝素(PHA)刺激(3天)人外周血单核细胞(PBM)中IL-2受体(p55, Tac肽)的表达,而不影响细胞的增殖。这种增强依赖于IL-2 (0-1 U/ml)和1 α, 25(OH)2D3(0-10(-7)M)的浓度。白细胞介素-1 (IL-1, 0 ~ 100 U/ml)的加入对IL-2存在的细胞中IL-2受体的表达没有增强作用。此外,1 α, 25(OH)2D3对两种细胞系Kit225(来自T细胞慢性淋巴细胞白血病患者的IL-2依赖性细胞系)和YT(来自急性淋巴细胞白血病患者的IL-2非依赖性自然杀伤(NK)样细胞系)具有相同的作用。因此,1 α, 25(OH)2D3增强IL-2对IL-2受体(p55)的上调,不仅在活化的T细胞中,而且在nk样细胞系中也是如此。
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引用次数: 0
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Nihon Ketsueki Gakkai zasshi : journal of Japan Haematological Society
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