The fixation methods and immunoperoxidase stains suitable for demonstrating both lymphoid cell surface antigens and Giemsa-stain morphology on imprint smears of lymph nodes were studied, with the following results.
1) The fixation methods for air-dried smears were:(1)35 % BFA, 30 sec at 4°C, (2) 25% PLPA, 10 to 15min. Both (1) and (2) were suitable for the purpose. BFA was more useful for preparing and preserving the fixatives than PLPA.
2) The PAB and ABC staining methods were both suitable.
3) Endogenous peroxidase activity should be blocked after washing of the secondaryantibody with methanol containing 0.3% H₂O₂ for 30min.
4) Unstained air-dried smears should be kept in a container with silica gel and stored at -18°C so that they can be stained after a considerable length of time.

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The effects of human embryonic fibroblasts (WI-38) on the morphology of a human scirrhous carcinoma cell line from a stomach cancer (KATO-III) were examined by light and lectron microscopy. When co-cultured with fibroblasts, KATO-III cells significantly increased in cell size and had vacuoles containing glycoproteins and acid polysaccharides. These characteristic changes in KATO-III cells were also observed when they were cultured with culture medium onditioned with fibroblasts. However, when KATO-III cells were cultured for another 7 days without fibroblasts, these characteristic changes gradually disappeared. These results suggest that humoral factor(s) secreted by ibroblasts had a reversible effect on the morphology of scirrhous carcinoma cells.

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The cell surfaces of the oral mucosa of denture wearers and non-wearers were examined for differences in ultrastructure. The subjects consisted of 10 denture wearers and 10 non-wearers, all of them healthy adult males or females. The hard palate and maxillary alveolarl ridge were selected as regions to be examined. The mucosa in all cases and exfoliative cells in some cases were observed in these two regions by scanning electron microscopy (SEM).
The results are as follows:
1. In SEM images of the mucosa, the ultrastructure of non-denture wearers included three types of microridges: the honeycomb type, the process type and the scattered micropits type, of which the honeycomb type was most frequently observed. With denture wearers, however, the honeycomb type was rarely observed. Instead, the scattered micropits type was dominant and the microridges of this type showed a tendency toward flatness. Also, in denture wearers, the curved type and straight type, which were not found in non-wearers, were noted.
2. In SEM images of exfoliative cells, conspicuous changes due to denture use were shown in keratinized and superficial cells of the hard palate and in intermediate cells of the alveolar ridge. With denture wearers, the honey comb type was decreased in keratinized cells, the scattered micropits type was increased in supercial cells and curves were increased in intermediate cells. These changes were almost identical to those in SEM images of the mucosa. However, in regard to the kinds of cells in other regions, their ultrastructural changes did not correspond to those in SEM images of the mucosa. A clear relationship was noted between each microridge type and the kinds of cells in terms of frequency regardless of whether or not dentures were used. The details are as follows: The honeycomb type appeared rather frequently in keratinized cells and parakeratotic cells. The scattered micropits type and process type were frequently observed in keratinized cells parakeratotic cells and superficial cells and less frequently in intermediate cells. The curved type, straight type and co-existence type tended to appear most frequently in superficial cells and intermediate cells. A comparative study of SEM images of exfoliative cells and the mucosa indicated possible differences in the ultrastructure of obverse and reverse exfoliative cells.
3. Ridge-like and fissure-like linear structures, which were observed in SEM images of exfoliative cells, are considered to be cytoplasmic impressions and not cell borders. The ridge-like linear structure was particularly revealed in SEM images of the mucosa.

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Fine needle aspiration cytology was performed in 99 cases of thyroid diseases during 8 years from 1980 to 1987 in Kurume University Hospital.
Among the 99 lesions, cystic change was observed in 32 cases.
The specimens for cytological and histological examinations were available in 19 cases (adenomatous goiter, 7 cases; Follicular adenoma, 4 cases; papillary carcinoma, 8 cases) out of the 32 cases with cystic change. In preoperative cytological examinations, atypical cells suggestive of malignancy were seen in 5 of the 8 cases of papillary carcinoma.
Thus, cytological examination of cystic fluid of the thyroid seems to be useful in the diagnosis of cancer.

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Intraoperative Douglas cavity washing cytology was performed in 387 patients with gastric cancer, of whom 243 had macroscopical serosal invasion. The relationship between the occurrence of intraperitoneal free cancer cells and stage, histological type, and histologically demonstrated depth of wall invasion was studied, and the prognostic value of the cytological features, and the number of intraperitoneal free cancer cells was investigated.
1. The rate of occurrence of intraperitoneal free cancer cells was 35% in the group with histologically demonstrated serosal invasion but 6% without it.
2. Poorly differentiated types of cancer showed a high rate of occurrence of intraperitoneal free cancer cells.
3. In well-differentiated types of cancer, the cluster formation type and the single scattered type of intraperitoneal free cancer cells were observed at almost the same frequency, but in poorly differentiated types of cancer, the single scattered type was dominant.
4. The influence of the difference in the cytological features of intraperitoneal free cancer cells on the outcome of the disease was not clear.
5. The outcome worsened as the number of intraperitoneal free cancer cells increased.

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DNA ploidy pattern of paraffin-embeded block from 67 patients with 10 benign, 41 malignant and 16 ovarian tumors of low malignant potential (LMP) were analysed by flow cytometry. Staining for DNA analysis was achieved with propidium iodide. All of 10 benign ovarian tumors exhibited only diploid cells. Tumor with aneuploidy or multiploidy pattern were found in 85.4% of carcinomas. Of the 16 tumors of LMP, almost tumors were diploid like adenoma, but only one was multiploid and had a episode of recurrence after 4 years interval.
Our results suggest that estimation of DNA ploidy pattern may be used as an additional diagnostic tool supplementing conventional histopathologic evaluation of ovarian tumors of LMP, but prolonged follow-up and increased patient accrual is necessary to assess whether the flow cytometric analysis will provide practical importance.

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A short-term estrogen test was used to obtain a correct cytologic diagnosis in 17 patients with an equivocal atrophic cellular pattern. Overestimation in cytodiagnosis was markedly eliminated and the correct cytologic diagnosis was made in 70% of the cell samples after estrogen test. Maturation of normal squamous cells was induced after the test. Drying artefact of the cell samples and cell aggregates were eliminated after the test.

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We investigated the relationship between the result of cytodiagnosis of peritoneal washings and other prognostic factors in specimens obtained from 42 patients with endometrial carcinoma. Cytodiagnosis was restricted to patients at stage I, II or DI in whom no extrauterine invasion or rupture of the coat was macroscopically evident upon laparotomy.
The percentage of patients with positive cytodiagnostic results was 16.7%(7/42), showing no obvious difference in the positive rate in relation to the histologic type or clinical stage. There was no difference in the positive rate between patients who had undergone hysteroscopy and those who had not. Among patients with myometrial invasion, no significant difference in the positive rate was found in relation to the depth of invasion, provided that the invasion had not reached the serosa.
The positive rate generally became higher as the proportion of the lesion extending to the uterus increased, some cases of small lesions showing positive cytodiagnostic results. Among patients in whom the lesion was distant from the tubal ostium, 20%(3/15) showed positive results. All 7 patients with positive results were given additional treatment and are presently alive without any sign of recurrence (after exactly 6 years at maximum, mean 3 years and 7 months).

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Findings of a gastric cancer with increased AFP, observed in a 71-year-old male, was examined by exfoliative cytology.
Two types of malignant cell clusters were distinguished in touch smear preparations of the biopsied gastric mucosa; one was adenocarcinoma cell clusters with the piling-up feature, and the other was one-cell-layer sheets composed of polygonal cells resembling that of hepatocellular carcinoma.
Histopathologically the gastric cancer showed a picture simulating trabecular hepatocellular carcinoma in almost all areas including small foci of tubular adenocarcinoma.
By immunohistochemistry, AFP, ALB, PALB and TF were detected in the cells of the first type of clusters.
A lectin-affinity test using immunoelectrophoresis revealed that 94% of the total AFP in the serum was associated with con-A, showing that the serum AFP of this patient was identical with that of ordinary hepatocellular carcinoma.

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A 76-year-old female was admitted to our hospital, complaining of epigastralgia. Gall stones and a cystic tumor of the pancreatic body were found on further examination. Aspiration cytology of the pancreatic cyst was performed during surgery. A moderate number of tumor cells loosely connecting with each other were found in the smears. Both tumor cells and their nuclei were round and of almost the same size. The nuclei had a smooth margin, and showed a fine reticular chromatin pattern. The cytoplasm of the tumor cells was faintly stained and had a blurred margin.
An enucleated pancreatic tumor measured 2.5×2.5cm in two diameters and had a cystic cavity containing bloody fluid and having a thin and yellowish friable wall. It was diagnosed as a non-functional glucagonoma, because almost all of the tumor cells gave a positive reaction with glucagon on immunohistochemical examination, but this patient did not present with either glucagonemia or symptoms.
Almost all cystic pancreatic tumors originate in the pancreatic duct and contain features of mucinous and serous types, but a small number of cystic islet cell tumors have been reported. Cells of islet cell tumors in cytologic smears are able to be differentiated from those of the mucinous type tumors, which show the presence of mucin in the cytoplasm, and from the serous type, which contain cytoplasmic glycogen granules, and tumor cells of both types appear in the smears and frequently form tightly connected cell clusters. These findings and those in the present case are thought to be of great value in the differential diagnosis of cystic tumors of the pancreas.

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Glucagonoma, difficult to find before operation, is a rare pancreatic islet cell tumor. A 71-year-old woman with glucagonoma which originated from the tail of the pancreas was reported. This tumor was diagnosed as benignancy that originated from a pancreatic islet tumor by preoperative aspiration cytology, and various preoperative examinations and clinical findings showed no evidence of glucagonoma. After operation, however, glucagon was shown by immunocytoand histochemical demonstration, and glucagon granules were shown by electron microscopy. And thus the tumor was fi nally diagnosed as glucagonoma. Consequently, the use of both aspiration cytology and immunocytochemical method will make it extremely helpful before operation to diagnose glucagonoma originating from pancreas, especially non-functioning case.

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Alveolar soft part sarcoma is an uncommon soft tissue tumor. We recently encountered an alveolar soft part sarcoma, primarily in the right lower leg. which was detected by the presence of a metastatic lesion in the left lung.
A 19-year-old woman was admitted to the hospital for evaluation of an abnormal chest shadow. A chest X-ray showed a round mass, 1.5 cm in diameter, in the left middle lung field. The patient underwent a left thoracotomy with removal of a well-circumscribed, yellow-white tumor measuring 2×2×2cm. A touch preparation yielded abundant cellular material. The tumor cells were present in loose groups or singly. The cells were relatively large and contained single or multiple eccentric nuclei, with a large nucleolus.The cytoplasm appeared cyanophilic. finely granulated. and on occasion vacuolated. Crystals were seen on PAS-stained preparations. Alveolar aggregates of the cells which were enveloped by a fine fibrous framework were readily found.
Histological examination revealed an organoid arrangement of cells separated by a thin fibrovascular stroma. The individual cells were large, with vesicular cytoplasm. Diastase-resistant, PAS-positive crystals were demonstrated. Immunoreactivity for NSE and S-100 protein was demon strated in the tumor cells. By electron microscopy, crystals a myelin-like structure were observed in the tumor cells.
An extensive search for the primary lesion disclosed amass in the right lower leg. This tumor mass was completely excised. A histologic diagnosis of alveolar soft part sarcoma was made.

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The clinical and cytological features of an inflammatory pseudotumor of the lung were studied. The patient was a nine-year-old boy in whom roentgenograms of the chest revealed a solitary circumscribed round mass, which was diagnosed as a granulomatous lesion by preoperative brushing cytology. As the mass increased slowly in size in spite of therapy for tuberculosis, an open lung biopsy was performed. In cytological specimens, short spindle-shaped cells tended to be arranged in a storiform pattern, and had basophilic cytoplasm, enlarged oval nuclei with slight atypia, thinner nuclear membranes, and a finer chromatin structure. These findings suggested their origin as epitheloid cells. Foreignbody-type multinucleate histiocytes were seen, but there was little necrotic substance and the background was clear. The diagnosis of an inflammatory pseudotumor was made by histopathological study. This lesion is a rare granulomatous lesion that must be distinguished from malignant tumors. However in none of the previously reported cases was the diagnosis established preoperatively. When brushing cytology of an intrapulmonary lesion shows histiocytic spindleshaped cells in a storiform pattern, but smears and cultures for tuberculosis are negative, particularly in children, it is necessary to examine cytological specimens carefully and take into consideration the possibility of an inflammatory pseudotumor.

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Spindle cell lipoma, proposed by Enzynger as a specific type of lipoma easily misdiagnosed as liposarcoma in 1975, frequently occurs in males at the age of about 50 to 70 years as tumorlike adipose tissue with occurrence predominantly in the occipital region, shoulder and back.
In the present study, we conducted fine needle aspiration biopsy cytology (ABC) for spindle cell lipoma on the back of a 48-year-old man and experienced difficulty in differentiating between benignancy and malignancy from the cytodiagnostic and histological aspects. The number of cells collected were enough to show a cellular mass consisting of mature fat cells with spindle-shaped nuclei. These cells were mixed with spindle-shaped cells with nuclei like a uniform short rod. Nucleoli were not prominent, but partially tachycromatic atypical nuclei were occasionally found.
Liposarcoma of the differentiated type could not be ruled out, so the tumor was classified as class III. No recurrence of the tumor was found 10 months after excision.
These observations cofirmed the diagnostic role of ABC for soft tissue tumors. The cytological and histological pictures of this tumor were discussed.

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A case in which mucinous adenocarcinoma of the ovary and carcinoma in situ of the uterine cervix were coexistent, was reported. A 53-year old woman visited our hospital because of abdominal distension and genital bleeding. The aspiration smears of ascites showed a great number of atypical clusters. Overlapping, small nucleus and hyperchromasia were observed.
Malignant cells contained positive materials for PAS staining. The cervical and endometrial smears showed same malignant cells, and many of them were not degenerated. The cervical smears showed small number of stripped cells.
The endometrial smears contained keratinizing atypical squamous cells. The histological postoperative diagnosis was double cancer: mucinous adenocarcinoma of the ovary with metastasis to uterine corpus and cervix, carcinoma in situ of the uterine cervix.

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