While I was at the department of surgery, Lund University, Sweden, for recent 5 months, I had an opportunity to yisit the aspirations clinic of university hospital. I describe brief summary of what I saw at the clinic. The cytology section belongs to the department of pathology, and most of the doctors working in the aspiration clinic are pathologists. The annual numbers of cytology cases during the last years have been increasing. The figures for 1977 are as follows: Vaginal cytology 43, 169, respiratory cytology and effusions 5, 364, urinary cytology 6, 704 cases and aspiration cytology 8, 574, whereof 4, 619 aspirations were made by the cytologists and the remaining cases were sent to the laboratory from other hospitals. An aspiration is performed by using a very fine needle attached to a disposable syringe, which is connected to a pistol-type aspiration apparatus. Aspirations cytology is performed at he outpatient clinic in most cases. Ten to 20 patients are received aspiration cytology every day. It is also frequently performed in the operating room and the fluoroscopy room. The number of cases by organs performed in 1977 is: 1, 969 cases of mammary gland, 1, 160 cases thyroid, 1, 004 cases of prostate, 1, 103 cases of lymph node, 513 cases of liver, and 344 cases of salivery gland cytology: thus, almost all the organs are included and cytology of the lung, pancreas and kidneys is also frequently performed. The clinicians in all the departments are greatly interested in and also greatly rely on cytologic diagnosis, utilizing the diagnosis for the treatment of various deseases, especially, for the early diagnosis and the determination of therapeutic regimen of cancer.
Our AUTO-CYTO-SCREENER, like all the other apparatus for automated cyto-screening, cannot always discriminate correctly malignant cells from other particles on smears, especially, neutrophile clumps. Smears in which cells are perfectly dispersed and equally scattered should therefor be constantly prepared to actualize automated cyto-screening. The special method of specimen preparation for this purpose has already been reported, but has not yet arrived at perfection. In this paper is reported further betterment of this method for preparation of smears for automated screening, which may be summarized as follows: 1) Vaginal smears for automated screening has so far been suspended and preserved in 50% ethanol according to our method. In this study, however, the preserving solution for smears was changed from 50% ethanol to 2% formaldehyde phosphate buffer (pH 7. 4), because coagulation of serum protein and aggregation of hemolyzed erythrocytes were caused by suspending smears in 50% ethanol, consequently impeding the exact measurement of cell numbers in suspensions with the blood cell counter. This phenomenon never occurred in the formaldehyde solution. 2) In order to disperse cell aggregates in smear suspensions, ultrasonic treatment has been adopted in our method, but has proved not so effective. A homogenizer was used for trial purposes to disperse cell aggregates instead of an ultrasonicator, and proved more effective than the latter. 3) It is earnestly desirable in putting automated cytoscreening to practical use to remove neutrophiles, erythrocytes or bacteria from smear suspensions. The use of Separate-L was attempted for this purpose. That is, 3ml of Separate-L (1, 067) in a test tube was overlaid with the same amount of a smear suspension. The loaded tube was allowed to stand without agitation for 3 to 5 hours instead of centrifugation. After this step, squamous epithelial cells in a smear suspension, including malignant ones, were concentrated at the bottom of the tube, while neutrophiles, erythrocytes and bacteria formed a band at the interface of the liquid layers. Utilization of this technique seems to be useful for preparing more excellent specimens for automatic cyto-screening purposes.
In order to obtain good specimens suitable for our automated prescreening machine for cervical cancer, Saccomanno's procedure being used for the detection of lung cancer was applied to the vaginal washings. The following procedure has been established: 10ml of vaginal saline washings is blended with a super high speed blender (Warning Timer-Light Blender, Scientific Products 8350 -1) at 21, 000 rpm for 5 sec. and fixed with an equal amount of 4% carbowax dissolved in 95%; ethyl alcohol for 30 min. and then centrifuged at 1, 500 rpm for 5 min. An appropriate amount of sediment is taken with a capillary pipet and homogeneously suspended in 4ml of 2% carbowax dissolved in 50% ethyl alcohol comparing the turbidity with that of Kingsbury-Clark's standard solution containing 10mg/dl protein so as to make the same turbidity. This solution is originally used for quantitative assay of urine protein. The suspension thus obtained is smeared with an automatic cytosedimentation machine (Sakura Autosmear, Model: CF-12SB) through 50μ square mesh filter at 1, 500 rpm for 5 min. The slides are allowed to dry completely. Subsequent Papanicolaou's stain is done by an automatic cytostainer (Sakura, Model: RSP-50). Following this procedure, 227 cases including 202 normal and 25 abnormal cases (dysplasia 8, carcinoma in situ 3, squamous carcinoma 13, adenocarcinoma 1) were examined and 126 good specimens (56%) were obtained. Good specimens were those which contained sufficient dispersed squamous cells for analysis without cellular clumps, excessive numbers of polymorphonuclear leukocytes or erythrocyte debris. Since the turbidity of suspensions was applied to the cell count, there were individual variations in the number of squamous cells. However, this procedure was simple enough for mass screening and excellent for cell dispersal. In addition, cells were sufficiently well preserved and showed no drying distortion as often observed in conventional Papanicolaou smears. In this study, Separate-L (Mixture of sodium metrizoate and ficoll) originally developed for separation of lymphocytes from blood was also applied to the vaginal washings of 26 cases to remove polymorphonuclear leukocytes which sometimes lead to misclassification of normal cells and was considered to be effective for 16 cases (62%). However, considering that this procedure is complicated and has a possibility for the concurrent loss of cancer cells, it requires further investigation.
Correct diagnosis of needle biopsy specimens of mammary disease cases based only cell morphology is frequently difficult. Therefore the author performed quantitative nuclear DNA measurement in mammary disease cases and investigated the relationship between morphological diagnosis and cytochemical aspects. The author investigated the relationship between the degree of histological differentiation, which reflects the degree of clinical malignancy, and nuclear DNA content. The material studied consisted of 125 cases; 30 cases of mastopathia, 30 cases of fibroadenoma, 7 cases of intraductal papilloma, 56 cases of carcinoma and 2 cases of sarcoma. Cells from imprint smears were used for nuclear DNA content mesurement. After morphological diagnesis of Papanicolaou stain specimens, Papanicolaou destaining followed by Feulgen staining was performed and DNA measured by microspectrophotometer. All benign cases indicated mostly diploid mode nuclear DNA content, with, on average, about 10% of cells above the diploid and up to the tetraploid region. However, there were two cases each of mastpathia and intraductal papilloma in which, albeit few in number, cells were observed beyond the tetraploid region. The findings of this study indicated that nuclear DNA measurement might therefore be a useful means for indicating those benign cases which require close follow up. Carcinoma cases displayed abnormal nuclear DNA content with distribution from the diploid to 5c modes, and many were observed with smaller peaks beyond the tetraploid region. Nevertheless some cases of well differentiated adenocarcinoma exhibited patterns not completely different from benign cases and therefore caution is advised in employing this method for definitive diagnosis. The result of this study revealed a relationship between the degree of histological differentiation of breast cancer and the nuclear DNA content can be applied to the estimation of degree of morphological malignancy.
In this report is present an examination of electron microscopic features of papillary thyroid carcinoma and benign and malignant breast tumors. There were many similarities between papillary carcinoma of the thyroid and papillotubular carcinoma of the breast in electron microscopic findings, but intracytoplasmic lumina were seen only in the carcinoma of the breast. There were many differences between benign and malignant breast tumors. The cells derived from the carcinomas indicated such features as intranuclear cytoplasmic invagination, coarse granular pattern of chromatin, large nucleoli, abundant intracytoplasmic organella and intracytoplasmic lumina. Among these features the intracytoplasmic lumina were considered significant findings in recognizing carcinomas in the breast.
Eight cases of bronchioloalveolar cell carcinoma were reviewed cytologically, and differential diagnosis conducted accordingly. There are two problems underlying differential diagnosis of bronchioloalveolar cell carcinoma. First, it is difficult to differentiate tumor cells from benign hyperplastic cells, especially in sputum cytology, owing to both the uniformity of tumor cells and the lesser degree of cellular atypism. Second, even if such cells are diagnosed as malignant, differentiation of these cells from the cells derived from other types of adenocarcinoma is problematic. Consequently, in an attempt to resolve these difficulties, the authors marked the shapes of tumor cell clusters by the bronchial brushing method. The shapes characteristic of cell clusters are presented below, and serve as a useful guide in cytological diagnosis of bronchioloalveolar cell carcinoma. 1. Single-layer cluster of cells along a one-dimensional plane, in which cell circumferences are smooth and/or sometimes dented. 2. Solid multi-layered cell cluster exhibiting distinct cell boundaries and abundant cytoplasma, conforming to a tortoise shell or honeycomb pattern. 3. Solid multi-layered cell cluster surrounded by amorphous mucin in which the cluster appears floating. 4. Solid multi-layered cell cluster conforming to a pattern similar to a bunch of grapes.
Cytological features of alveolar cell carcinoma in sputum and direct method obtained by four patients were described and the source of exfoliated cancer cells in the airway discussed from the pathohistological standpoint. The cytological characteristics of alveolar cell carcinoma in that smear material consisted of a cluster formation of carcinoma cells with tight cellular junction and the rounded and smooth outer margin of the cell cluster. Each cancer cell was columnar or cuboidal, the nucleus was eccentric, the nucleoli were small, and the nuclear chromatin did not marked increase. The cancer cells were uniform in shape. As exfoliated alveolar cell carcinoma clusters were observed in the normal alveolar speaces, it was considered that exfoliated cells were expectorated outward from the route via normal alveoli.
Smear examinations were performed on 31 oral lesions of squamous cell carcinoma. Cytologic diagnosis was positive (Class IV, V) in 21 (67.7%), suspicious of malignancy (ClassIII) in 5 (16.1%) and false negative (Class I, II) in 5 cases (16.1%). Histologic sections were classified into 5 groups, based on the degree of differentiation or some variants. The cytologic specimens were correlated with the histologic findings, and those in the false negative cases were also studied. The results may be summarized as follows: 1. Clinically, leukoplakic lesions show less malignant characteristics in cytologic examination; however, erythroplakic, erosive or ulcerative lesions generally exibit a positive cytologic diagnosis. 2. Histologically, in the cases showing severe atypical changes and frequent mitotic figures, positive cytologic diagnoses were obtained easily. It was also shown that the more dedifferentiated the cells, the more were lost the characteristics of squamous epithelium. We can therefore diagnose poorly differentiated cases as malignant by cytology, but it is difficult to confirm the histologic type. 3. We think that cytology of oral lesions has limitations according to our technique, and that further studies are needed to improve its reliability.
In this study it was examined by using Yoshida Sarcoma (YS) cells what was the most suitable method of preparing specimens of the pure pancreatic juice obtained through a cannula using a duodenoscope. The following are the experimental conclusions and their applications. 1. When ethylalcohol was diluted with distilled water, YS cells were well preserved for 5 days at room temperature in a more than 50 per cent solution of ethylalcohol. 2. When ethylalcohol was diluted with Hanks solution, YS cells were well preserved for 30 minutes even in a 25 per cent ethylalcohol solution in an ice bath. But the cells became small in proportion to the concentration of ethylalcohol. For this reason the pancreatic juice was collected in an equal volume of 50 per cent ethylalcohol in an ice bath. 3. Usually, the pancreatic juice is collected with a contrast medium (Urografin) after endoscopic pancreatography. The contrast medium (76% Urografin: osmotic pressure 51.9 at., specific gravity 1.422) made YS cells small and obscure, and also made centrifugal cell concentration difficult. To avoid the loss of the cells, we use the nuclepore filter technique. And at the time of screening attention needs to be paid to small cells. 4. To avoid the distortion of the cell with the nuclepore filter, the positive pressure must be kept at a minimum.
Among human neoplastic tumors in gynecology, uterine sarcoma is a very rare malignancy and accounts for 1.1 to 5.5% of all malignant uterine tumors. Uterine sarcomas arise from the muscle, connective tissue, vascular elements, endometrial stroma, and others. In the case of poorly differentiated sarcoma, it is difficult to diagnose the type of sarcoma. However, there should be little difficulty in diagnosing leiomyosarcoma by the use of specific stains such as PTAH, van-Gieson. and Mallory. We attemped to use these specific stains in smear cytology and were able to diagnose two leiomyosarcomas. The cytoplasm was stained light green by Papanicolaou and contained fibrous materials. The fibrous materials were stained deep blue by PTAH, yellow by van-Gieson and red by Mallory. The histological appearances in the 2 cases were interpreted as leiomyosarcoma. These cellular details in smear cytology by specific stains well corresponded to the histological features of leiomyosarcoma.
A case of bilateral breast tumors in a 33-year-old woman with complaints of a tumor mass in the left breast and bloody discharge from the right nipple is reported together with the results of cytological and light microscopic examinations. The tumor of the left breast was 4×6cm in size, well demarcated, and diagnosed clinically as fibroadenoma. In cytology of the touch smears of the resected tumor were found distinct malignant cells, and the specimens of the resected tumor were diagnosed histologically as “Noninfiltrating papillary carcinoma”. The smears of bloody discharge from the right nipple which was free from tumor mass showed the following features: the cells were arranged in clusters; the cytoplasmas were 9-24 microns in size; the nuclei were 7.5-18 microns in diameter, round or oval in shape, and had fine or coarse granules. These cytological features were judged as malignant. But the histological features of the specimens obtained by the simple mastectomy revealed atypical intraductal papilloma and/or duct papillomatosis (borderline).
A case diagnosed by sputum cytology as hepatoma is reported. Two kinds of carcinoma cells were observed. One comprised very atypical and like adenocarcinoma cells, and the other, those not so atypical but containing cytoplasmic droplets, and granular pigments which were stained green by Papanicolaou stainning. It was confirmed by autopsy that these cells were hepatoma cells. Well differentiated hepatoma cells can be diagnosed cytologically, and intracytoplasmic bile pigments are pathognomonic for hepatoma.