Experimental and clinical studies were carried out to investigate the optimum conditions of the washing solutions in lavage cytology under direct vision by fibergastroscope for the diagnosis of gastric neoplasms. 1) The effects of various washing solutions on preservation of morphology and function of gastric cells were examined experimentally by using cytological, electronmicroscopic and cytochemical methods on exfoliated gastric cancer cells. Hanks' solution was the best among 5 kinds of washing solutions. 2) Effects of various washing solutions on viability and morphological changes of HeLa Cells were investigated. Hanks' solution at an optimum pH 7.4, gave the most excellent results. Effects of osmotic pressure were not significant. 3) When pH of washing solutions was less than 6.0, marked degeneration of cells was observed. Phosphate buffer was more effective in buffer action, but degeneration of cells was more remarkable in phosphate buffer than in Hanks' solution. Excellent results were obtained by adjusting the pH of Hanks' solution to 8.0, when pH of gastric juice was less than 4.0.
[A] Three methods are used at present in direct vision cytology; the lavage method, the suction method, and contact smear of biopsy tissue. This paper is to discuss the clinical value of these techniques with special reference to the suction method. The instrument used for suctioh method is the Model B fiberscope (fiberscope for biopsy), with which a thin Teflon tube with a beveled metal tip instead of the biopsy forceps is introduced into the stomach and advanced, under direct vision, to the given target. Repeated negative pressure using a 100 ml syringe is then applied to suck material into the tube. The material is then flushed and smeared on a slide glass. This method is characterized by the following, 1. Preliminary lavage is not necessary. 2. Abundant fresh cells can be obtained with minimal necrotic debris. 3. Multiple lesions can be examined simply by changing the tube. 4. The damage given to the tissue is least in comparison with the lavage method or biopsy. This method is, therefore, particularly suitable for minute lesions, but is difficult to obtain material from deeper portions. 5. The suction method has given correct diagnosis in 97% of the cases, that is comparable to the other methods. The method thus suffices clinical purposes, but the result will be further improved if these techniques are combind acording to the location or morphological characteristics of the lesion to be examined. [B] By the morphological analysis of exfoliated cells of gastric cancer, by examining the touch smear specimen of the biopsy tissue, the following results were obtained. The general features of the tumor cells of early gastric carcinoma, in comparison with those of advanced gastric carcinoma, were as follows. 1. The tumor cells of early gastric carcinoma had poor anisocytosis, and were smaller in size. 2. Chromatin was thin and poor in early carcinoma cells. 3. Nucleoli were smaller in early carcinoma cells. 4. In advanced gastric carcinoma mitosis was found more frequently than in early carcinoma. 5. Glandular formations were seen more ofen in early carcinoma specimen. Accordingly, in the aspect of clinical cytology, the exfolliated cells of early gastric carcinoma have lower malignant signs than these of advanced gastric carcinoma.
1. Basic Experiments As a model experiment to analyse the malignant behavior of tumor cells, two kinds of transplantable rat ascites hepatoma cells (AH-62F and AH-66F) were investigated by a follow-up study on the metastic processes, and its results were compared with surface property of tumor cells which was examine by the colloid and Methylene Blue absorption method, and by cell electrophoresis. Marked differences were observed in biological properties between these two hepatoma cell lines; AH-66F showed a prodominent malignant behavior like short survival of the host rats and higher frequency of tumor formation in various organs and lymph nodes after interavenous ingculation However, frequency in liberation of tumor cells into the thoracic ducts did not vary greatly in both cases, in which tumor cells either of the two lines were inoculated into the colonic mesenterium. A marked difference was also found in electrokinetic properties between the two lines of these ascites hepatomas. The cell surface of AH-66F was constructed of substances containing electrically charged substances of higher density and particulary of sialic acid-dependent electrical charge, which was confirmed by measuring the decreased rate of electrical charge density of cell surface after treatment with sialidase. Based on these results, the electrokinetic properties of tumor cells was discussed and compared with their malignant behaviors. 2. Electrokinetic property of human gastric cancer cells and their histopathological characteristics. The electrokinetic properties of human gastric cancer cells and all the surrouding mucosal cells were investigated comparatively by using cell electrophoresis. Cells were obtained from surgical specimens by treatment with collagenase and hyaluronidase immediately after their surgical removal. In all 20 cases, gastic cancer cells showed higher electrophoretic mobility than the mucosal cells. In the differentiated cases of gastric cancer, individual tumor cells were all uniform, while wide individual difference in electrophoretic mobilities was noted in the anaplastic cases of gastric cancer. In particul the latter cases showed higher avarage value in electrophoretic mobility than the formor.
An attempt was made to demonstrate the presence of tumor specific antigens in cancer cell of the human endometrial adenocarcinoma and to apply it to the diagnosis of malignant cells in endometrial smear using immunofluorescent antibody technique. Rabbits were divided into 3 groups. The different groups were immunized with nuclear, mitochondrial and microsomal fractions prepared from cancer cells of endometrial adenocarcinoma by the method of W.C. Schneider. The sera obtained from each groups were absorbed by the each fractions prepared from normal epithelial cells of the endometrium. Their specifities were determined by the Ouchterlony plates. On agar gel diffusion tests, tumor specifities were shown in antimitochondrial and antimicrosomal sera. In the immunofluorescent staining for tissue sections of endometrial adenocarcinoma and aspiration smears, the indirect technique of Coons was carried out using the sheep anti-rabbit globulin antiserum conjugated to fluorescein. On the histologic sections and aspiration smears prepared with antimitochondrial and antimicrosomal sera, the specific fluorescence was seen in the cytoplasm and a few granules in the nucleus of cancer cells. On the contrary, no fluorescence was seen in normal endometrial cells. From above mentioned facts, it is evident that the cancer specifity existed in mitochondrial and microsomal fractions, and this specific fluorescence can contribute largely to detection of cancer cell in aspiration smears and endometrial tissue.