The in vitro colony formation by marrow from 16 children with normal bone marrow and 27 children with acute leukemia (16 cases at onset or relapse, and 11 cases during complete remossion) has been studied. The number of colonies per 2×10⁵ cells in normal bone marrow ranged between 126 to 504, and the average count was 259 colonies. This was a higher number of colonies than that in adult bone marrow. After 7 days of culture, colonies were macroscopically recognized as white spots. The ratio between compact colonies to loose colonies was approximately 2:1, when classified using an inverted microscope.
In 12 cases which had counts in excess of 95% of blasts at onset or relapse of acute leukemia, there was a decrease in colony formation. One case of ALL showed an almost normal colony forming capacity. All of the 4 cases with 30 to 60% of blasts developed colonies, but the number of colonies was less than that of normal bone marrow. Colony forming capacity in 11 cases with remission was similar to normal bone marrow. We consider that in some cases of acute leukemia at onset or relapse there may be blasts that can differentiate and maturate.

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体盤幅1mmと5mmのアラスカカスベBathyraja aleuticaの胚のパラフィン連続切片を作成し, それらの組織発生を調べた.材料は, 1982年に東部ベーリング海大陸斜面の海底域からトロール網によって漁獲された卵殻内より採集されたもので, 主として, 始原生殖細胞の出現部位とリンパ組織の発達を観察した.体盤幅1mmの胚で, 神経管, 脊索, 体節, 原始腸管, 中胚葉側板などが観察されたが, 器官形成はまだ行われていなかった.始原生殖細胞は, 体壁中胚葉及び内胚葉と内臓中胚葉の間にある間葉に存在していることが分かった.一方, 体盤幅5mmの胚では, 形成過程にある多くの内臓諸器官がみられた.始原生殖細胞は, 前段階の部位には認められず, 生殖隆起表層内に集合していた.二対の胸腺原基は咽頭域に存在しており, リンパ球で充満していた.また, 多数の未熟な血液細胞からなる脾臓原基と思われる小型の組織が認められた.

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As it is well known, the abdominal lymph nodes are influenced by food. But this fact has not yet been shown on the cells and tissues of the lymph nodes of rats. Through the per os administration of D. A. B.[p-Dimethylaminoazobenzene (Butter yellow)]in olive oil a hypertrophy of the abdominal lymph nodes, resulted, increasing large cells in the sinuses and parenchyma. The giant cells seemed to form by the fusion of a number of cells. The large cells and the giant cells appeared unexpectedly not only through the administration of D. A. B., but also of olive oil as well as of high molecular palmitic acid. Through non-fat food, giant cells appeared in the parenchyma, but the large cell did not increase in number in the sinuses.
The large cells are phagocytic. This was seen after the injection of Indian ink or YOSHIDA sarcoma cells in the peritoneal cavity. The large cells, which had been too much loaded with erythrocytes, presumably could no more protect the invasion of the sarcoma cells. The large cells are in their multiplying period, judging from the behavior of the mitochondria, in full activity.
The above-mentioned large cells in the sinus are derived from reticular cells of the sinus, by losing the reticular fibres. Most of the giant cells in the parenchyma seem to be producted by the fusion of the large cells.

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Tlae existence and the distribution of the Nymph vessels in the t?amstertestis were studied morphologically in detail. By the perfusion metlaod for fixation, theNymph vessels were observed in the interstitial tissue just adjacent to the seminiferotustubules with tire light and electron microscope.The penetrating of the Nymph vessels through the albuginea testis was confirmed andas a result of tracing these Nymph xessels, the coarse net works of the Nymph capillarieswere reconstructed and figured. The lymphatic net works were recognized to anastomoseeach other and their branches were situated just close to the seminiferous tubules and tlaeinterstitial cells. In the electronmicroscopic observation, these Nymph capillaries sltowedthe closed s)stem and so their free endings were not opened.The tissue spaces had no direct communication to the Nymph capillaries but they weresuggested to form the sinusoidal pathways of the same protein-rich liquid as that in theNymph vessels, and the important role of the interstitial tissue spaces as the passages orextracellular liquid were discussed.

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Reciprocal interaction between the hemopoietic organ stromal cells and the cells of the granulocytic, megakaryocytic and erythrocytic series in the human liver obtained from 109 embryos 28 to 49 days after ovulation and 76 fetuses from 8 to 22 weeks of gestation were investigated by light and electron microscopy.
The close association of stromal cells with immature cells of the three series was confirmed under the electron microscope and a presumptive HIM (hemopoietic inductive microenvironment) was visualized. A majority of immature erythroblasts intruded into the cytoplasm of the hepatocytes, so the presumptive hemopoietic stem cell types II and IV are undoubtedly differentiated into cells of the erythroid line by contact with hepatocytes at a certain stage of maturation. Granulopoiesis developed among the reticular cells around the ductus venosus—or large arteries in hepatic parenchyma—and the cells of the granulocytic series were enclosed by thin cytoplasmic projections of mesenchymal cells. Neither erythropoiesis nor megakaryopoiesis was noted here. Therefore the compartments composed of one or more reticular cells around the ductus venosus or large arteries seem to have a capacity to regulate the differentiation of the presumptive hemopoietic stem cells type IV into cells of the granulocytic series. This differentiation of presumptive hemopoietic stem cell types II and IV into the megakaryocytic series is belived to be induced by the presence of the microenvironments that consist of foci of a few reticular cells in the hepatic parenchyma, as immature cells of megakaryocytic lineage were encircled by the cytoplasmic projection of one or more reticular cells among hepatocytes.
Erythroblastic islets are concluded to be a kind of HIM, where erythroblasts loosely adhere to the central macrophages and undergo mitoses and maturation.

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Two EBV-negative human B-lymphoma cell lines, BJAB and DG75, were transfected with an Epstein-Barr virus (EBV) nuclear antigen 2 (EBNA-2) gene, which plays a critical role in the EBV-induced immortalization of primary B lymphocytes. Furthermore, DG75 cells were co-transfected with the EBNA-2 gene and a latent membrane protein (LMP) gene. Expression of eight surface antigens on the resultant EBNA-2-expressing cell clones was analyzed by flowcytometry. None of the EBNA-2-expressing cell clones derived from BJAB and DG75 showed a significant increase in the expression of cell surface marker CD23, of which enhancement by EBNA-2 in a different EBV-negative human B cell line, Louckes, was previously reported. Expression of CD25 (IL-2R/Tac) on cell surface, however, was induced in two of six DG75-derived cell clones. One of the two CD25-induced cell clones was expressing EBNA-2 only, and the other was co-expressing EBNA-2 and LMP. The results suggest that EBNA-2 has a potential to up-regulate CD25 independently of CD23 on human B cells.

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Histological changes of the skin inoculated with vaccine virus by means of the sand paper method are observed fromimmune biological point of view. (See P. 108)
1) After 5 hours of this vaccination, epidermis cell changes to necrosis and the neutrophilic infiltration appears in demarcation zone. When these tissues discharged after 5 days, there prepared already regenerated epidermis beneath the scab. However, these changes are seen in the contrast experiment by simple sand paper rabbing or electric burning. So the charactericity of the vaccinated epidermis should be represented by the regenerated epidermis in the following stage.
2) The inclusion body appears in the cell body of the regenerated hypertrophic epidermis and the follicule epithelium after 2 days and till 7th day. After 4 days a part of the hypertrophic epidermis is liquidated calling the infiltration of the neutrophilic cell and postulating the locus. Following this reaction the monocytic infiltration appears in the papilla, growing gradually to the depth of the corium along the hair follicule and reaching to the reticular layer.
3) After 8 days the hemorrhagic inflammation appears in the monocyte infiltrated corium which makes this leisions necrotic and causes to fall off. After this time neither the inclusion bodies nor the monocytic infiltration were to be found. These findings show that there is a close correlation between the existence of the inclusion body and the monocytic reaction in the epidermis. It can be said that the tissue reaction against the vaccinia virus is the monocytic nature.
4) The colour of the neutral-red vacuoles of the histiocyte in the reticular layer was observed supravitall, which changed to brownish during the period of 3 to 17 days after the vaccination in parallel to the alkalosis which occured the inoculated tissue.
5) The plasma cells began to grow close to the addventitia cells in the subcutaneous tissue 4 days after the inoculation. After 7 days they infiltrate in cluster and after 8 days explosively around the area of the hemorrhagic infiltration. The proliferation of the plasma cell occurs on the period of alkalosis of the inoculated locus as well as the bone marrow successive to the leucocyte infiltration and alkalosis of the tissues.
6) 20 to 25 days after the primary vaccination the reinfection experiment was applied to the abdominal side of the animals (for the observation of the Pirquet's immediate reaction). The erythema and the swelling of the inoculated locus reached to its maximum after 24 hours transitorily without any following pustulation.
7) The neutrophilic reaction on the vaccinated locus at the beginning of the second inoculation appeared more intense than that of the primary, somewhat like the mode of Arthus Phenomenon.
8) As the destruction of the virus was performed by this Phenomenon so neither the inclusion body nor the monocytic infiltration was to be seen in this reinoculation. However, the call of the plasma cell reaction is rather quickened.
9) pH of the locus is alkalified in parallel to its erythema and then returned to the normal state in a short time. However, the brown colour of histiocyte-vacuoles is still remained after the recovery of the tissue pH, which means brown colour of histiocyte-vacuoles is not influenced simply by the tissue pH.

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Cancer in the genitourinary tract is the increasing problem. The lymphatic system, which plays an important role in metastasis is still obscure and vast field is left for investigation.
This paper presents animal and clinical study of the lymphatic flow, utilizing radioactive colloidal gold ¹⁹⁸Au.
(1) Animal study; after subcutaneous injection of ¹⁹⁸Au on the dorsal part of dog's foot, uptake of radioactivity of various organs was investigated by scientillation counter, rate meter, well counter etc.
Thoracic duct canulation was performed and the uptake correlation between thoracic duct lymph and peripheral lymph node was investigated, using accelerationg and delaying factors as well as obsering the effect of anethesia and death.
(2) Clinical study; 104 cases (normal 85/abnormal 19) were investigated for femo-inguinal and iliacal nodes uptake after injection of ¹⁹⁸Au to dorsal part of feet.
a) 6 hours after injection, all of them (104/104) had some uptake, proved by scintillation counter.
b) 1 hour after injection, only 2 out of 57 without accelerating factors, 19 out 19 with them in normal legs, compared to 0 out of 4 without them, 3 uou of 3 with them in edema legs had uptake
3) Urological study (comparison of lymphangiography and 198An lymphscanning) 38 urological and gynecological patients were investigated.
Abnormal scintiscanning found in 2/3 in kidney and adrenal tumor, 1/1 in giant tumor in the pelvis, 4/4 in penis and genital tumor, 1/7 in bladder tumor and 1/13 in uterus carcinoma.
Though my study is still limited, I feel that ¹⁹⁸Au is useful for investigation of lymphatic flow and its clinical urological study can be applied to broad field.

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