Thrombopoietic (Tpo) and megakaryocyte-colony stimulating (Meg-CSF) activities were found in urinary extracts from patients with aplastic anemia. Preparative biochemical extractions were accomplished using Sephadex G-50 and DEAE-cellulose column chromatography. The biological activities of these extracts were assessed using not only an in vivo assay but were also examined in vitro employing the clonal development of megakaryocyte colonies. Both in vivo, as well as in vitro, biological activities were detected in the batch fraction which was stepwise eluted from DEAE-cellulose between 0.022 M NaCl in 0.016 M NaH2PO4 and 0.15 M NaCl in 0.05 M Na2HPO4 as a single fraction. When 0.4 mg of this fraction was injected daily into rats, a marked thrombopoiesis ensued producing an increase of 40% over initial platelet counts by 3 days after administration. This was followed by a decrease in platelets to a subnormal range by 21 days after the injection. Hemoglobin concentration gradually increased from 5% above initial value by day 7 to 20% above initial value by day 21. The effect of neuraminidase (NAse) on the properties of this extract was also examined. NAse-treated extracts, similar to the native extracts described above retained Tpo activity. Changes in megakaryocyte numbers in the spleen and bone marrow of rats were assayed with both the NAse-treated extract as well as with the native extract. A remarkable increase in megakaryocyte numbers, threefold above the normal count, was found in the spleens of rats given the native extract preparation; by contrast, however, no change was observed in splenic megakaryocyte numbers in rats given the NAse-treated extract. On the other hand, NAse-treated extract retained its ability to stimulate bone marrow megakaryocyte proliferation in the same rat. The urinary extract also revealed in vitro Meg-CSF activity with a specific activity of 31, 750 CFU-Meg colonies/mg of protein.

中文翻译：

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从再生障碍性贫血患者尿液中提取的血小板生成素的部分纯化和生物学特性。

在再生障碍性贫血患者的尿液提取物中发现了血小板生成（Tpo）和巨核细胞集落刺激（Meg-CSF）活性。使用Sephadex G-50和DEAE-纤维素柱色谱法完成制备性生化提取。这些提取物的生物学活性不仅使用体内测定法进行评估，而且还利用巨核细胞集落的克隆发育进行了体外检查。在分批级分中检测了体内和体外的生物学活性，该批级分从DEAE-纤维素中逐步洗脱出0.016 M NaH2PO4中的0.022 M NaCl和0.05 M Na2HPO4中0.15M NaCl中的单个级分。每天将0.4 mg的该级分注射到大鼠中后，出现明显的血小板生成，给药后3天比初始血小板计数增加40％。随后在注射后21天，血小板减少至低于正常范围。血红蛋白浓度从第7天高出初始值的5％逐渐增加到第21天高出初始值的20％。还检查了神经氨酸酶（NAse）对这种提取物性能的影响。与上述天然提取物相似，NAse处理的提取物保留了Tpo活性。用NAse处理的提取物和天然提取物均检测大鼠脾脏和骨髓中巨核细胞数目的变化。使用天然提取物制剂的大鼠脾脏中巨核细胞数量显着增加，是正常数量的三倍。相比之下，在用NAse处理的大鼠中，脾脏巨核细胞数量没有变化。另一方面，NAse处理的提取物在同一只大鼠中仍具有刺激骨髓巨核细胞增殖的能力。尿液提取物还显示了体外Meg-CSF活性，比活性为31，750 CFU-Meg菌落/ mg蛋白。