Employing 99 rabbits, white and male, in size about 2kg., conducted a femur marrow puncture, and by pouring Indeanink and Berliner blue into a femur marrow vessel, immediately after the operation until 100th day, made some transparent specimens, casts of vessels by means of injection of Resin, as well as H. E. sections for the purpose of investigating the regenerative process of the bone-marrow, especially to know the unique property of sines nets of the marrow; results were as follows: Further, I studied on destruction and regeneration of 3 corpses that had underwent a sternal puncture before death. (1) After the puncture. that part remains, the cavity of which has been constructed owing to the destruction, filled with blood; yet it happens seldom that such thick vessels as the principal artery or principal sinus should be broken. (2) The regeneration of sinus nets, begins on 3rd day, roundabout the destructed part; the invasion of regenerative vessels into a part of destruction becomes gradually strong from about 4th day. Moreover, their regenerative conditions look like the sporting or budding of trees, and seen to formulate come sinus nets by anastmosis of one regenerative sinus with another, or a regenertive sinus and completed sinus. (3) Though small in number. the regeneration of arterial capillaries are seen in the destructed part on about 4th day of operation, which gives birth to anastmosis with some regenerative sinus nets in the neighbourhood, or already completed ones. But no anastomosis between arterial capillaries could be detected. (4) It took about 7-12 days for an unfinished atypical sinus nets that were thin and variously-formed to cover the whole destructed part. (5) Most of the regenerative marrow vessels near the destructed part of the bone were derived from those vessles due to endoperiost, and due to anastomosis with regenerative vessels in the bone marrow. (6) The so-called intersinusoidal capillaries are seen not only under the feeble state of bone-marrow function, but also in the regenerative process of the bone-marrow vessels. (7) The regenerative faculty, or the regenerative function of bone marrow vessels mainly depends upon age of those test rabbits; it has proved much stronger in young rabbits than in old ones. (8) In case the marrow-punture is done continually for a short time, clinically, care should be taken to except at least over 20 mm distant place from the previpus one.
Using 49 test rabbits to observe both the destruction and regeneration of their marrow-tissue, by dint of H. E. section, as well as print preparation of the bonemarrow, obtained results as follows: (1) The marrow part aspirated and demolished by the marrow puncture has been filled with clots of blood as well as necrotic tissue; but it has to be replaced comparatively quick by some connective tissue cells originated from normal marrow tissue around the destructed part and endoperiost; then, it became regenerative marrow tissue rich in fat cells which, in turn would recover its proper state. (2) Most of the histiocytes that have been producted within the regenerative marrow tissue having cleaned that part, have yeilded to regression and degeneration, and until to disappearance. However, a part remained as it was. (3) The giantcells, which would appear after the regenerative marrow had been cleaned due to histiocytes, compared to normal marrow tissue, have proved a denser distribution for a while, but disappeared, leaving a part. (4) The fact that we can detect owing to print preparation of the bone marrow is that hyperemia is detected due to aseptic inflamative process, the multiplication and regeneration of interstitial cells are seen at first, next, it turns to multiplication of parenchymal cells.
I have taken some investigations on the manipulation of intramedullary transfusion, along with on the effects of the transfused blood on bone-marrow vessels, parenchyma, as well as the peripheral blood picture; then, further about the fate of blood used for the blood transfusion. (1) It has been observed that sinus nets have increased its capacity to an maximum extent, immediately after the blood transfusion, centering on a part of blood transfusion, so that the diameter of sinuses has become several times larger that of normal one; though it is evident that certain sinus nets would never succumbed to change. The destruction-ends of sinuses owing to marrow-puncture have already been blockaded immediately after the operation. Some sort of recovery on the part of vessels was taken place in an area destructed in the bone marrow, due to blood transfusion; which probably would cost it 7 days or so until the entire regeneration has been achieved. No intersinusoidal capillaries could be detected in those sinus nets. (2) Though such sinuses as the collective sinuses which were not affected by the marrow puncture, or the principal sinus that were not subjected to ordinary obstruction, have increased their volume after the transfusion on a sudden, no self-destruction occurred; which, it would cost 2 or 3 days to recover normality; in some cases, even after 5 days, entire recovery could not occur. (3) Arterial capillaries have been influenced in no wise at the time of puncture, except they suffered the destruction; even in that circumstance, their destraction-hole would be closed in two days lapse. Moreover, no marked change occurred to the position of arterial capillaries and capacity, under the blood transfusion. (4) Almost no vessel destruction would happen to both the paralell arterial branches or the principal artery, due to the marrow puncture, and transfused blood; besides, neither the positions nor the capacity proved change at all. (5) A marked appearance of nucleated erythocytes were detected in peripheral blood after the blood transfusion, but it would decrease in paralell whith the regeneration of sinus nets. so, after 5-6 days, no trace has been detected. (6) The left shift of nuclear is wont to cotinue till 7th day from the operation, at leucocyte picture, arriving at the climax 2 or 3 days after. Also, the leucocytes stood in a correlation with those pseudoeosinophilic lucocytes which, even if would decrease for a while after the blood transfusion, increases on the spot, regenerating gradually into the normal state in 24 hours. (7) As to the fate of transfused blood, a part would invade directly into sinuses at its end, then went out of the body; while other part, though existing for a while between the sinus-wall and the parenchyma, in gradual stage demolish sinuswall, shifting into the sinuses and out of the bone-marrow. Further, that part of transfused blood destructed in parenchyma has turned to Hemosiderin, and been absorbed as foreign matter.
Studies on reverse anaphylaxis and influence of various medicines for growth by means of tissue culture. (1) The reverse anaphylaxis were achieved by means of tissue culture. (2) Adding complement is necessary to cause reverse anaphylaxis. (3) It needs, at least and enoughly, 20 minutes for complement activity to be gottenon. (4) Forssman's antigens were included in hens embryo and membrane of blood vessel. (5) Penicillin, dyhydrostreptomycin and merzonin could inhibit the growth of bacteria. (6) The higher concentration of medicine inhibit the growth of tissue. (7) Megabion accerate the growth of tissue, if added a drop at the concentration of 10-610-7 and yet megabion could not coagulate plasma by itself.
The production of antibody in blood was studied by tissue culture, and the results were as follows. (1) The blood antibody is produced in leucocytes. The production of antibody is serum>leucocytes>defiblinated blood in descending order. (2) The appearance of precipitin and agglutinin in cultivated tissue proves serum>lymphnode>leucocytes>defibrinated blood in descending order. (3) The liofilized chick embryo juice could be stored aseptically. The activity of growth accelerater in chick embryo tissue juice does not decrease for two months.
Of course tissue culture is best method to apply for experimental reserch on organs producing antibodies. The author, on late having been informed of the recent importance of antibodies production due to lymphocytes, conducted certain experiments about it: results were as follows; (1) The productions of hemagglutinin, hemolysin, Forssman's antibody, bacterioagglutinin and precipitin were recognized in pure cultivated lymphocytes. (2) Antibodies producted in pure cultivated lymphocytes had specifity against corresponding antigens. (3) The productions of antibodies in lymphocytes reached the highest titer more rapidly compared to the titer of antibodies in blood, but decreased more rapidly too. The maximum titer of tissue antibodies proved higher than that of blood antibodies. (4) It was recognized that precipitins are made rather in a slower degree than that of other antibodies. In case immunized antibodies prove great in quantity serum antibody value proves higher, but in case it is small in amount antibody production supersedes that of serum.
While the researcher was studying on the heterogene reaction in the serum protein precipitation and the anaphylactic phenomena, especially on the heterogene reaction in the anaphylactic reaction from the view-point of changes of the generations, he observed the heterogene reaction in the serum protein precipitation in Part I, the heterogene reaction in the anaphylactic reaction from the view-point of change of the genations when seen in the case of anaphylactic shock, the drop of temperature, and the anaphylactic intestinal contraction in Part II; the heterogene reaction in the anaphylactic reaction from the view-point of changes of the generation when seen in the cases of anaphylactic shock; a drop of temperature and anaphylactic intestinal contraction; the heterogene reaction in the anaphylactic reaction from the view-point of changes of the generations when seen in the case of changes of blood images, and those of pulmonary tissue. And he is going to summarize the results of his experiments as follows; 1. In the anti-serum whose antigen is cow serum and every fraction of the serum protein, its precipitation titer and precipitation gehalt are in proportion to the frequency of injection, to which dose the heterogene reaction to the serum of other animals run parallel. The subject of heterogene reaction is contained in the euglobulin and pseudoglobulin or fraction of globulin. The specific singularity of albumin is very strong. 2. According to the experiment on the anaphylactic reaction of a guinea pig, caused by the anaphylactic shock, the anaphylactic intestinal contraction, the change of blood images, and that of lung tissue, etc. when cow serum is used as sensitizing antigen, anaphylaxis changes through generations and in the case of a child of the second generation, is produced not only by the chief antigen cow serum, but also by such the side antigens as goat serum, pig serum, horse serum. Moreover in the experiment on anaphylactic intestinal contraction. it can be noticed that anaphylaxis changes through generations in the case of a child of the third generation, too. 3. When cow serum albumin is used as sensitizing antigen, nothing but the reaction to the chief antigen cow serum and the side antigen goat serum can be noticed, and the specific singularity of albumin is strong, and consequently it can be confirmed from the view-point of changes of the generations that the subject of the heterogene reaction in anaphylaxis is contained in serum euglobulin and pseudoglobulin, that are fractions of globulin.