Distribution of zinc in the human body and its metabolism in neoplastic patients have long been a subject of intensive study, and numerous reports are available in this respect. Recently McNary has reported a new histochemical method of identification of myeloid granules in the human leucocyte using dithizone as staining agent, and the method was applied by Szmigielski as a means of cancer diagnosis. Several other researchers have recognized some advantage in this method for its relative simplicity and reliability. As part of the study on the zinc metabolism in the cancer patients that has been conducted in our laboratory, the present author made a detailed study on the method of leucocyte staining by dithizone and results obtained were presented in the followings. In view of the situation, the present author undertook to invenstigate into the details of the leucocyte staining procedure by dithizone from initial step of blood smear preparation and laking of erythrocyte to various conditions such as room temperature, time required for air drying and staining, pH optima of the staining solution and the complex-forming buffer. The present studies on such basic problem concerning the possibility of analyzing the trace amount of zinc in leucocytes have proved successful in establishing a reliale staining procedure of myeloid granules in leucocytes with the use of dithizone, and evidences were presented that the granular bodies stained in bronze color by the procedure were exclusively of zinc. Zinc was decreased significantly in granulocytes in patients of stomach cancer (12 cases) but not in patients of oral cancer (4 cases) when examined by staining solution from which the complex-forming buffer was omitted.

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Studies were made on serum protein fractions and serum iron levels in 33 patients with oral cancer and in the rabbits which were implanted with Vx2 carcinoma in the oral region. Antitumor agents were administered to the patients and the experimental animals by intra-arterial infusion. The serum protein fractions and the iron levels of serum before and after the infusion were analyzed for their possible significance as diagnostic aid of cancer, and at the same time observation was made on the effect of the antitumor agents upon the progress of the tumor lesions. Determination of serum protein fractions was carried out by electrophoresis on cellulose acetate membrane and iron was determined by Matsubara's method. Results obtained were summarized as follows. 1. Total protein, albumin fractions and iron level of the patients serum before administration of antitumor agents were slightly lower than healthy controls and patients bearing oral tumors of benign type. Levels of alpha 2-globulin and gamma-globulin were respectively slightly higher in the cancer patients than in the healthy controls. 2. A significant difference was found in serum protein spectrum and iron level between patients with cancer of maxillary sinus and patients with maxillary sinuitis, and these data seemed to serve as an aid in differential diagnosis. 3. The decrease of serum albumin, increase of alpha 2-globulin and gamma-globulin and decrease of serum iron level paralleled well to the progressive stages of cancer lesion. 4. No significant fluctuation in the serum patterns was observed throughout the period of medication except some irregular serum patterns temporarily observed. 5. Significant improvement in the serum patterns was obtained after administration of antitumor agents in most cases of patients where the drugs were effective, and the lesser the effect of drugs, the poorer the improvement, and no such improvement was shown when the drugs were totally ineffective. 6. The intra-arterial infusion given to healthy rabbits caused slight decrease of serum total protein, albumin fractions and iron level and slight increase of beta-globulin. 7. Administration of antitumor agents to the experimental animals bearing Vx2 carcinoma by way of intra-arterial infusion often resulted in improvement of the serum patterns, but failed to do so by way of intravenous injection of the agents. Excellence of intraarterial infusion to intravenous injection of the antitumor agents was further confirmed by far extended survival time of animals given intra-arterial infusion of the drugs as compared to those given the drugs intravenously. Improvement of the serum patterns was correspondingly noted in the former group of animals but not in the latter group.

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To approximate as nearly as possible to clinical condition for fractionated intra-arterial chemotherapy in oral cancer chemotherapy KB cells of 100x10^4/10ml LY-BSS cultivated at 37℃ for 3 days were exposed for a 30 or 60 minutes period at 37℃ to each of the following antitumor agents such as Nitromin (1mcg/ml, 10mcg/ml), Endoxan (2mcg/ml, 20mcg/ml), Toyomycin (0.01mcg/ml, 0.1mcg/ml), Mitomycin C (0.04mcg/ml, 0.4mcg/ml) and Bleomycin (3mcg/ml, 30mcg/ml) prepared with LY-culture medium containing 10% inactivated bovine serum. The antitumor agents were then washed out with Hanks's BSS. Then new LY-culture medium was added to the cells and kept at 37℃ for 3 days. Then the effects of antitumor agents from standpoint of viable cell number, morphologic changes, succinic dehydrogenase activity were studied. Subculture to another culture flasks was carried out simultaneously. Forgowing operation was repeated 5 times. The results obtained were as follows : 1. The results in inhibition of propagation of cells : Effect of Toyomycin varied with concentration and period of treatment. Nitromin and Mitomycin C had nearly the same tendency in inhibition of propagation in cells but they were affected more by time than concentration of treatment. Endoxan and Bleomycin showed different results from above three agents and it was rather affected by concentration than time of treatment. 2. No significant difference was apparent between any two agents in morphologic changes. However, slight tendency for degeneration of cytoplasm was observed in Nitromin ; regressive changes of nucleolus in Mitomycin C ; multinucleated cells in treatment of 20mcg/ml for 60 minutes in Endoxan, compared with other agents. In addition, segregation, cohesion in chromatin, pycnosis, atrophy, fragmentation as regressive changes in nuclei, atrophy in nucleolus, cytolysis were somewhat more noticeable when propagation of cells was inhibited. 3. Giant cells, microcells were observed in each of experimental series but types of agents, time, concentration and number of treatment did not cause any significant difference. The number of giant cells appeared was higher in lower concentration of agents. Also regressive changes of nuclei in giant cells was stronger in higher concentration of agents. Somewhat more microcells were observed in Bleomycin compared with other agents. 4. Increase and decrease in mitosis nearly agreed with increase and decrease in cells in Nitromin and Toyomycin but did not obtain so close an agreement in Mitomycin C. A lot of mitosis and aberrant division were observed through each of experimental series in Endoxan and Bleomycin compared with other agents. 5. No significant difference in changes of succinic dehydrogenase activity was evident depending on type of agents, time, concentration and number of treatment. The changes, however, had a correlation with increase and decrease in cells.

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The purpose of this investigation was 1) to analyze horizontally the growth changes of dentofacial pattern in mandibular prognathism before treatment, and to evaluate the influence of treatment by comparing the mean changes occured during treatment periord with the general tendency of the growth changes, and 2) to detect the factors influencing the severity of mandibular prognathism, and to find the differences of response to treatment among mandibular prognathisms of various intensity. Two hundred and thirty-one cases including seventy-two males and one hundred and fifty-nine females, ranging from age 4.67 years to age 27 years, of anterior cross bite and mandibular prognathism were selected for roentgeno-cephalometric study. These cases were classified into five age groups and four skeletal types, by males and females respectively, according to the chronological age and the apical base relationship before treatment. All these cases showed negative overjet before treatment and showed positive overjet after treatment. The lateral headplates taken before and after treatment were used for tracing and analysis. Areas studied were relative to the craniofacial skeletal pattern and the denture pattern. Several findings were observed : 1) The flexion of the cranial base (∠BaSN) showed no significant difference among various age groups in the average before treatment, and revealed no significant change after treatment. 2) Dimensional increment of S-N, S-Ba, Pns-Ans, Go-Me, and Ar-Gn during treatment periord were regarded as contribution of the growth. 3) In the skeletal pattern, ∠SNA, ∠SNAns, ∠PP-SN, gonial angle, Ar-Go, and Ar-Ba remained stable during treatment period. 4) No evidence of growth activation on anteroposterior dimension of the maxilla was noted although maxillary dentition had been treated on the basis of labial expansion. Dimensional growth of the mandibular body and effective length of the mandible did not seem to be retarded in spite of the fact that the retraction force had been applied on the mandible during treatment period. 5) The position of the mandible was found to be shifted backward and downward as relative to the cranial base under the influence of the treatment. Skeletal discrepancy between the maxilla and the mandible was found being corrected to a certain degree mainly by this positional change of the mandible. 6) Axial inclination of the incisors was found to be affected most markedly by treatment. The maxillary incisor was tipped labially and the mandibular incisor was tipped lingually by the treatment. 7) Dimension and the flexion of the cranial base did not seem to be correlative with severity of mandibular prognathism. Undergrowth and retrusion of the maxilla seemed to play only a small part in the severity of mandibular prognathism. 8) However, summation of the dimensional factors and positional factors of the mandible would affect the severity of mandibular prognathism more conspicuously. 9) Contrary to the expectation and the ordinary concept, it was found that the cases of severer mandibular prognathism would tend to have a smaller gonial angle and a smaller mandibular plane angle in the average. 10) It was revealed that the more the mandible protruded, the more the maxillary incisor inclined labially and the more the mandibular incisor inclined lingually before treatment. The tilting up anteriorly of the occlusal plane and the increasing of the overbite would associate with this. 11) The more the mandible protruded before treament, the more it would be retruded by the treatment. It does'nt necessarily mean that the mandible was easier to be retruded by the treatment when it had been more protruded. It would rather mean that it was more necessary to retrude the mandible in the severer mandibular prognathism although there exist a limitation of the retrusion. 12) The more the mandible protruded before treatment, the less the maxillary

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For the purpose of pursuing cytological effects of anticancer drugs on oral malignanttumors, mitomycin c and bleomycin were injected intraarterilly and intravenously into rabbit ear transplanted with Vx2 Carcinoma. The author took daily smear from transplanted tumor, and observed gross and microscopic findings. In clinical study, intraarterial infusion of anticancer drugs alone was used for 16 cases. Those treated by radical operation and/or irradiation were excluded from this study. Smear taken daily for study of exfoliated cytology and prognosis. The following results were obtained. 1. The form changes occuring in the smear obtained by anticancer drugs, were also observed in experiment of rabbits and clinical cases, these changes were thickening, resolution and extinction of nuclear rims, paling of staining, prominency, resolution and extinction of nucleoli ; in cytoplasm, vacuolation, paling of staining, indefinite border, enlargement of the cell. 2. In the experiments of rabbits, exfoliative cytological effects were observed 4th or 5th injection after intraarterial infusion of mitomycin c, and 3rd for bleomycin. Effect occured after the 5th for intravenous infusion of mitomycin c and bleomycin. During the period, constraction of tumors was observed. 3. In clinical cases, cytological effects were observed after administration of 20 amples of mitomycin c by intraarterial infusion ; 10 amples of bleomycin. During the period, constraction of tumor and ulcer was observed in many cases. 4. In experiments of rabbits and clinical cases, cytological effects of anticancer drugs were observed earliest for bleomycin intraarterial infusion. 5. These cases showing higher results in experiments and clinic in cytological diagnosis also showed eminent effect in histopathological diagnosis and prognosis were good.

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