癌患者の血中銅および亜鉛量に関する研究 : 第 1 編 白血球分離に関する研究

抄録

Since McNary reported a new method for staining leucocytes using dithizone and Szmigielski applied the method as a cancer diagnosis, many researchers studied on the zinc content in the leucocytes of the tumor-bearing organism. But the methods applied in these reports are mainly qualitative ones, and quantitative studies have been quite seldom. In the field of oral surgery, only the studies by Fujimoto (1970) and Tamari (1971) can be encountered, and these remain also histochemical, and therefore qualitative. In order to determine the zinc content of leucocytes as accurately as possible, several methods hitherto reported for leucocytes separation were reexamined, and a new method suitable for the purpose have been developed. The procedures and the results obtained with this method were summarized as follows. Ten ml of blood was drawn with the disposable syringe which had been heparinized previously, and centrifuged at 700 G for 10 minutes in a polyethylene tube. Then the buffy coat and its neighboring layer (3 ml), which were rich in leucocytes, were aspirated with a siliconized pipette, transfered into TERAOKA's centrifuge tube, a special tube constricted at the middle portion, and centrifuged again at 230 G for 10 minutes so that the leucocytes were allowed to deposit in the narrow portion. The leucocytes thus collected were transfered into a 50 ml centrifuge tube with a siliconized capillary pipette, and the erythrocytes herein intermixed were hemolysed by adding 0.28% saline solution. After further centrifugation, the supernatant was removed, and the precipitates were washed with 0.9% saline solution and suspended in a total of 3 ml of the saline. The suspention was incubated at 37℃ for 30 minutes, and then the number of the leucocytes was counted. In the final suspention as prepared by this procedure, recovery of the leucocytes from peripheral blood was 56.8% on an average, and the ratio of erythrocytes to leucocytes was as low as 1 to 5, and 90% of thrombocytes was removed. A separate experiment also confirmed that even the ghost of erythrocytes remaining after hemolysis contained no more than a trace of zinc. Thus, the contamination by the remaining thrombocytes, erythrocytes, and their ghost would seem almost negligible with this procedure. It may be added that hematological composition of the isolated leucocytes was not different from that of the peripheral blood. One more advantage is that the procedure made it possible to estimate the amount of copper and zinc in the plasma as well as in erythrocytes with use of a single blood sample.