Exact knowledge on cancer cell cycle is important for cancer chemotherapy. However a large quantity of purified cancer cells is necessary for measurement of nuclear DNA contents with Flow cytometry (FCM). And it is possible to measure nuclear DNA contents of clinical small specimens with Micro-spectrophotometer (MSP).
We disigned to study usefulness of MSP to assess cell cycle compared with FCM. We measured nuclear DNA contents in six kinds of human cultured cellslung zancer (adenocarcinoma), hepatoma, bone origine giant cell tumor, rhabdomyosarcoma, neuroblastoma, and Raji cell, and investigated correlation between DNA histogram and cell doubling time.
The results are summarized as follow;
1) Nuclear DNA histogram was seprated into two components-lst component containing G₀G₁ and a part of S phase, and 2nd component containing other part of S phase and G₂M.
2) A significant correlation was found between two 2nd components with MSP and FCM (r=0.914, p<0.02).
3) S phase was detectable with MSP if large number of cells entered S phase.
4) A negative correlation was found between 2nd component with MSP and cell doubling time (r=0.925, p<0.01).
5) These data indicated that it was possible to assess cell cycle of cancer cells with MSP, and this was seemed to be important cytologically.

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Fundamental studies were performed on the detection of IgM in the mouse spleen lymphocyte with the view of application of immunoperoxidase technique to clinical cytology. As for sampling of cells on the glass slide, especially in imprint smear, the important factor is uniform scattering of cytological specimen on the slide, excluding tissue fluid which may contain any kinds of antigen.
For this purpose, we washed out completely the cytological specimen with 1% BSA in PBS in order to remove the liquid component containing extracellular antigen (wash-out method), wet fixation was superior to dry one in two points; clear findings of positive reaction and well-preserved cellular features with minimum swelling and deformation of cells. Sufficient fixation did not reduce immunoreactivity, but gave satisfactory cytological preparation.
Best results were obtained by fixation of the wet smear with methanol-ether (1:1) for 30 minutes, and the dry smear with aceton for 30-60 minutes at 4°C. The attempt of digestion by proteolytic enzymes (pronase P and trypsin) enhanced immunoreactivity, without significant reduction of background staining. In general enzymatic digestion is to be avoided, in dry preparation, since it brings about more or less cellular degeneration, even in for short time incubation and with low concentration of the enzyme. Application of surface active agent such as tween 20 and 80 could not reduce background reaction, but increased specific immuno-reaction and enhanced endogenous peroxidase activity of the smeared cells. Among the immunohistochemical procedures, indirect method seemed to be most appropriate, because of easier technique and less background staining, so long as and-serum titers are high enough.
Simple PAP method showed almost the same stainability as the indirect peroxidase labelled technique applied to enzyme-digested specimens, although the former resulted in slightly increased background reaction.
The most favorable results were given, when the PAP method was applied to washed-out preparation above-mentioned.
These results could contribute to further application of immunoperoxidase technique to clinical cytology.

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In vitro drug sensitivity of several cell lines of cultured human cancer cells against anticancer agents, Mitomycin C and Carboquone, was studied and compared from the view point of morphological changes of cells temporally exposed to the test drugs. The results and conclusions follow.
1. Morphology of the cells exposed to a certain amount of the test drug which has a certain biological effect on the cancer cells, i. e. 90% lethal dose, was not uniform but varied according to length of exposure time.
2. When the exposure time was fixed, the chains of morphological changes of the cancer cells correlated not to their drug sensitivity, but mainly to the growth rate of the test cells.
3. These results lead to the conclusion that drug sensitivity of cancer cells can not be judged on a morphological basis.

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Pulmonary hamartoma is one of the common benign tumors in lungs. Differentiation of hamartoma from malignant lung tumors by chest X-ray is sometimes very difficult. Therefore, it is of clinical value to make a cytological diagnosis of the hamartoma before peration.
For this purpose we studied a pulmonary hamartoma cytologically with materials obtained by needle biopsies (percutaneous, during operation and postoperative). Fourteen resected pulmonary hamartomas including twelve chondromatous and two non-chondromatous hamartomas were utilized for this study.
Cartilage cells, epithelial cells, fibroblasts, histiocytes, lymphocytes, polymorphonucleocytes and fat cells were found in the smears. Cytological findings well reflected histological characteristics of pulmonary hamartomas. In diagnosis of the hamartoma, it is important to recognize two types of cells cartilage and epithelial. Cartilage cells which had a small irregular nucleus with gray or reddish purple cytoplasm were found in all cases of chondromatous hamartomas. Epithelial cells were found in eighteen out of twenty-one materials. These epithelial cells resembled the bronchial epithelial cells, but in some cases showed a slight atypical changes. Ten of twelve chondromatous hamartomas were diagnosed cytologically.
In conclusion, a pulmonary chondromatous hamartoma can be diagnosed by cytological analysis of the smears obtained by needle biopsy.

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Bronchial brushing cytology was performed in 20 patients with pulmonary tuberculosis.
The cells obtained by this procedure included epithelioid cells, Langhans' type giant cells, hyperplastic columnar epithelial cells and squamous metaplastic cells.
Epithelioid cells were found in 55% of the cases, but Langhans' type giant cells in 5%.
Squamous metaplastic cells were found in 75% and hyperplastic colmunar epithelial cells in 20%, respectively.
Epithelioid cells were present in 80% of patients having positive tubercle bacillus in bronchial brush specimens or sputum obtained after bronchial brushing.
Based on our studies, it was confirmed that presence of epithelioid cells in bronchial brush specimens strongly suggest the presence of pulmonary tuberculosis.

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Atypical cells in sputum from 10 cases of the common cold with severe cough were analyzed. These cases were found out of 2840 sputum specimens procedured in Saccomanno's cell concentration method modified by Sato. These cells were seemingly misinterpreted as malignant, but they were not seen after remission of the symptom. The further observation suggested their origin of the degenerated bronchial columnar epithelial cells. Respiratory infection is thought to accelarate the degeneration and desquamation of the bronchial colmnar epithelial cells.

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Under certain circumstances, relatively large lymphocytes which have irregular shaped nuclei and are inversion of the nucleus-cytoplasm relationship appear with very high frequency in the pleural cavity effusions of some patients with malignancy or chronic diseases. They proved to be T lymphocytes by morphological, immunological and cytochemical examinations. Proportions of T lymphocytes of the pleural cavity effusions were extremely higher than those of peripheral blood. However thoracic duct lymph contains almost 100% T lymphocytes. T lymphocytes of the pleural cavity effusions may leak from the thoracic duct and/or branches because of contraction, occlusion, or injury of the thoracic duct as a of result pressures of malignant tumor etc.

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To study the relation of cytologic findings to histologic features in gallbladder diseases, aspiration cytology of the gallbladder using its contents aspirated during surgery and histologic examination of the resected gallbladder were performed in 101 cases with cholelithiasis. On cytologic examination, evidence of epithelial cells, inflammatory cells (leukocytes, lymphocytes and histiocytes) and debris were investigated. The resected gallbladder were histologically classified into normal, mild cholecystitis, lymph-follicular cholecystitis, acute obstructive cholecystitis, chronic obstructive cholecystitis and cancer.
The relation observed between cytology and histology follows; Only scattered clusters of epithelial cells were seen in normal cases, and epithelial cells, a few inflammatory cells and debris in mild cholecystitis. In lymphfollicular cholecystitis, the smears showed lymphocytes-the aggregates of which were occasionally diagnostic-added to epithelial cells and debris. Numerous inflammatory cells, especially leukocytes and histiocytes, debris and single epithelial cells were exfoliated on the dirty smears in acute obstructive cholecystitis. A few clusters of epithelial cells, debris and a few multi-nucleated giant cells were found in chronic obstructive obstructive cholecystitis. Only one of four cancer cases revealed cancer cells.
In conclusion, the relationship between cytology and histology of the gallbladder with moderate or high inflammation is present, but no sufficiently accurate cell collection is obtained to make a cytologic diagnosis possible through aspiration in normal and cancer cases. It would be a problem to be resolved in future to achieve such accurate cell collection through aspiration.

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In order to compare the detection rate of uterine cervical neoplasia in two scraping methods, spatula and cotton tipped applicator, 4, 255 women were screened using both methods simultaneously at Wakayama Rosai Hospital from 1978 to 1980.
With the spatula scraping method, ten cases of frank invasive cancer, fifteen cases of microinvasive cancer, twenty three cases of carcinoma in situ and twenty three cases of severe dysplasia were detected. On the other hand, with the cotton tipped applicator method, ten cases, fifteen cases, twenty cases and fifteen cases were found respectively.
A significant difference was found in the detection ratio of carcinoma in situ and dysplasia: that of carcinoma in situ was 87.0% (20/23) with the cotton tipped applicator method and 100% (23/23) with the spatula scraping method, and that of severe dysplasia was 69.6% (16/23) and 100% (23/23) respectively. There was no difference in the detection ratio of invasive cancer (10/10) and microinvasive cancer (15/15).
Accuracy of cytology with both methods was studied in 96 cases operated for uterine cervical cancer in our hospital.
Accuracy of cytology in frank invasive cancer was 81.2% with the cotton tipped applicator method and 81.0% with the epatula scraping method, respectively 61.9% and 75.0% in microinvasive cancer, 69.6% and 80.6% in carcinoma in situ, 48.2% and 79.2% in severe dysplasia.
The spatula scraping method proved more accurate than the cotton tipped applicator method in cytological diagnosis of uterine cervical neoplasia.

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The authors discovered two different atypical cells while conducting sputum cytology on an 81-year-old male. Because each cell had the features of small cell carcinoma cells and squamous cell carcinoma cells, we believed this to be a case of a primary double pulmonary carcinoma of small cell carcinoma and squamous cell carcinoma. The subject died in six months, and we had the opportunity to perform autopsy, which confirmed the presence of two types of carcinoma. It is extremely rare for a patient to be diagnosed as having double carcinoma while still alive. The major part of this study involves the cytological findings while the subject was still alive. The outline of the histopathological findings upon autopsy is also given along with discussion of the literature on primary double pulmonary carcinoma.
The authors also point out that the possibility of discovering such cases through cytological observation will increase in the future.

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A 62-year old woman was admitted to Miyagiken Seijinbyo Center for examination of a mass lesion in her left breast. The lesion was proved positive for malignancy and was diagnosed as scirrhous carcinoma by aspiration cytology. The patient underwent simple mastectomy.
The tumor, measuring 30×25 mm, was diagnosed histopathologically as infiltrating lobular carcinoma. Smear preparations of the resected material, treated with Papanicolaou staining, revealed following cytolo gical characteristics. Abundant cancer cells were scattered solitarily or arranged in small clusters comprising less than ten cancer cells. Indian filings were noticed. There were 29 pairs of cells (pair cells) out of 300 cells. Most cancer cells were small. The long and short axes of nuclei distributed in the range of 6-11μm (mean 8.83μm, S. D. 1.07μm) and of 5-10μm (mean 7μm, S. D. 1.02μm) respectively. Shape of nuclei was basically circular, but nuclear indentation was recognized in 29 out of 300 nuclei. Nuclear chromatins generally showed finely reticular or finely granular pattern, but coarsely granular chromatins were partly seen. Most of the cancer cells presented only one nucleolus of 1μm in size.

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Since 1978 we are performing cytologic study of the inflammatory bowel disease (IBD) for the early detection of its malignant change. An adenomatous precancer lesion in a patient of Crohn's disease of 20 year duration was detected by cytologic examination.
A 38-year-old woman was admitted to the Institute of Gastroenterology Tokyo Women's Medical College because of bloody diarrhea and abdominal pain. She has been diagnosed as ulcerative colitis at the age of 20. At the first admission to our institute in 1975. a diagnosis of Crohn's disease in the ileum and distal colon was made. After her discharge medical treatment, chiefly oral administration of SAS, continued. In 1979 a flat mucosal elevation was noticed in the rectum by colonoscopy. Brushing cytology of the focus was evaluated as strongly suggestive of malignancy. Eight months later the focus became more distinct and diagnosed as marked dysplasia by biopsy. In May 1980 the involved ileum and left-side colon were resected. A definitive diagnosis of Crohn's disease was settled. The focus in the rectum was 22×16 mm in size and diagnosed as marked dysplasia of adenomatous type (Riddell).
Cytologic study of the focus was carried out by colonscopic brushing in 3 times and imprinting smear of the surgical material. Papanicolaou and Giemsa staining were applied and the former was used for the measurement of the nuclear size. For the comparison, cytologic specimen of ulcerative colitis (46 cases) and colonic cancer (6 cases) were investigated.
The nuclear size of the cells taken from the focus was 16.7μ in the average and bigger than that of large bland cells or large active cells (Galambos) in ulcerative colitis. Anisonucleosis was more marked. The nucleoli were observed in 78% of the cells and more prominent than that of the latters. The cellular features of the former more resembled to that of the colonic cancer cells.
Since malignant-looking atypical cells are often exfoliated in IBD, the cytologic diagnosis of its malignant change is supposed to be very difficult. However presented case has clearly demonstrated the usefulness of the cytologic examination. Multiple biopsies, more than 10 for each segment of the colon, is recommended for the eraly detection of premalignant lesions in the patients with long-standing IBD. We believe that brushing cytology may be more suitable for this purpose.

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In a 24-year-old woman with hepatosplenomegaly and anemia, Gaucher's disease was indicated in aspiration cytology of the bone marrow and diagnosed by β-glucosidase activity in lymphocytes.
On examination of Giemsa-stained and/or Papanicolaou-stained smears, Gaucher's cells were large, and were round or polyhedric shape.
PAS reaction and acid phosphatase reaction were stronger positive in smaller cells than in larger cells. Sudan III stain was negative.
Ultrastructurally, the most characteristic appearance was tubular structures within a single membrane. These tubular structures were straight or serpiginous, 200 to 500 Angstroms in diameter. Lysosomes and autophagosomes were found, although quantity varied greatly from one cell to another. Erythrocytes were often and granulocytes rarely seen in the cytoplasm of Gaucher's cells.
It had been shown that Gaucher's cells carried receptors for C3 and Fc. The presence of Ia-like antigens on their cells were also indicated.
These cytological, cytochemical, ultrastructural, and immunological results showed that Gaucher's cells might be macrophages.

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In this paper a case of a 45-year-old man with bizarre leiomyoblastoma arising exogastrically was reported. Cytologic finding of touch preparation was not characteristic. The cytoplasm was mostly round or polygonal sometimes fibrous and no clear perinuclear spaces. The nucleus of this tumor was round or oval, the nuclear rim was smooth, and no mitosis was found. The tumor cells had abundant cytoplasmic filaments under the electron microscope.

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We have experienced a case of testicular tumor, which was suggestive of malignant lymphoma through the cytological examination of hydrocele fluid.
The patient, 74 year-old man, was admitted to our hospital becouse of painless swelling of the testis. Orchiectomy was performed with histopathological diagnosis of malignant lymphoma, histiocytic type. Generally establishment of preoperative diagnosis of a testicular tumor is not easy and correct diagnosis is usually obtained by histological examination of the orchiectomized specimen. At this point, cytological examination of hydrocele fluid of the testicular tumor is very useful in early preoperative diagnosis. Furthermore we reviewed the literature of the other testicular tumors, compared with malignant lymphoma, and discussed the cytology of the hydrocele fluid.

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A case report of malignant melanoma arising from male urethra was presented, urinary cytological material and stamp smear from the tumor was examined in addition to ultrastructural study of the recurrent tumor of the melanoma.
The patient was a 70-year-old male. Complaining of pollakiuria, dysuria, and hematuria, he visited the clinic of urology. The urethral tumor was found and histopathologically diagnosed as malignant melanoma.
Many tumor cells were observed upon urinary cytology after operation. Most of the tumor cells were solitary and scattered. Large nuclei with prominent nucleoli were eccentrically located in the cytoplasm. The cytoplasm contained a small number of melanin granules. The cells scanty in melanin granules were similar to those of a adenocarcinoma.
Ultrastructurally, the tumor cells had numerous melanosomes in the cytoplasm which were mostly type 1 and 2, with some of type 3 and 4.
The intranuclear vacuoles stained with light green for Papanicolaou's staining were often seen, but under electron microscope the vascuoles were thought to be intranuclear invagination which was well defined with a nuclear membrane, they contained many intracytoplasmic organellae and melanosomes.
In this case, we thought urinary cytological examination might be important before the operation, because the probe biopsy of malignant melanoma was contraindicative.

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1. A cytologic and clinicopathologic analysis of 6 patients with vulvar extramammary Paget's disease was reported.
2. The disease was seen predominantly in the older age group: The median age at onset was 65.7 years. Clinically, the lesion appeared erythematous (4 patients) to pigmentary (one patient), and all patients complained of itching. One patient was found to have the tumor formation into the vulva.
3. A close association of the Paget's disease with cancer was evident by considering that in addition to extramammary Paget's disease of the vulva, 3 of 6 patients had primary lesion in multiple locations; two patients had underlying adenocarcinoma and one patient had squamous cell carcinoma in other lesion of the vulva. Two patient had downward growth of the tumor from the epidermis proper. There were thus only one patients who had Paget's cells confined within the epidermis.
4. The prognosis of this disease was not so good, because one patient died of coexisting underlying adenocarcinoma of the vulva and two other patients had local recurrence.
5. Cellular smears obtained by scrapping and contactpressure technique from the vulvar lesion revealed unsatisfactory cytologic detail and only two cases before surgery showed a positive results, although moderate hyperkeratosis was present in 4 of 6 cases before primary surgical therapy. But we had a positive cytological diagnosis in all of two patients with local recurrence after surgery.
6. The cytological characteristics of vulvar extramammary Paget's disease was similar to that of Paget's disease of the nipple such as described by others. Abnormal cells were characterized by enlarged nuclei, both central and peripheral in location, with an increased nuclear/cytoplasmic ratio, granular nuclear chromatin with abnormal distribution, and prominent nucleoli. A lack of squamous keratinization was common to all specimens. In two cases, “cell in a cell” and “crescent formation” were observed. A small amount of melanine granule was found in the cytoplasm in one case, which indicated the necessity to differentiate extramammary Paget's disease from malignant melanoma.

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A 29 y. o. female patient (gravida 1, para 1) was contined atypical genital bleeding since puerperal. About 4 months after puerperal when she developed tumor on chest wall and cervical tumor formation. According to histopathological findings of above tumors, which diagnosed as Lymphosarcoma. This patient died at about 8 months after puerperal. Vaginal smear cytological studies (Papanicolaou Stain) were made of above rare case of cervical Lymphosarcoma.
In the result of above studies, the tumor cells are characteristically slightly larger than normal small lymphocyte, round forms and met merely differences of shape, with scanty cytoplasm. The nucleus are characteristically slightly variable in size and shape but some are round to oval while others are wedgeshaped and cleaved formation. And the nucleus revealed pale, increased of fine granular or rough granular chromatin, the circumference of nucleus manifested thick and the number of nucleoli tends to be low (1 or 2) and basophilic.
By means of above findings, this case was diagnosed as mid cellular type (B cell type) of diffuse cellular lymphoma owing to the classification of LSG.

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A case of rhabdomyosarcoma of uterine corpus was reported. Pure rhabdomyosarcoma of uterine body is a rare disease which is classified to pure heterologous sarcoma.
There were 50 cases of pure rhabdomyosarcoma of uterine body reported in the literatures. Among these 50 cases a few cases had achieved 5 year survivals. Our case is now living with N. E. D. after 5 years from initial treatment, and this is a very rare instance of this sarcoma.
The pathological and cytological specimens of this case revealed that there were many round or tape shaped or racket shaped rhabdomyoblasts in both of the specimens. Typical cross striations were seen in the malignant cells of elongated type in the pathological specimens. After careful observation, same striations could be found in the cytological specimens.

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