An investigation has been made on the intracellular localization of tumor specific antigen of the cervical cancer by means of immunofluorescent antibody technique. The cancerous and normal tissues obtained from the patients of cervical cancer and from the normal cervices were fractionated by the method of W.C. Schneider into nuclear, mitochondrial and microsomal fractions respectively. Rabbits were divided into 3 groups and immunized with each fraction. The antisera vs. each fraction of cancerous tissue were absorbed by the each fraction prepared from the normal cervical tissue. The specificities of these sera were determined by the Ouchterlony plate. On agar geldiffusion tests, the tumor specificities were present to anti-mitochondrial and anti-microsomal sera. Histological sections and vaginal smears taken from squamous cell carcinoma of the uterine cervix and from normal cervix were stained by Coons' indirect immunofluorescent method using anticancer mitochondrial and microsomal sera. In this test it was found that specific fluorescence was shown in the cytoplasm of cancer cells and no fluorescence was seen in normal cervical epithelial cells. Consequently, it is evident that the cancer specificity exists in mitochondrial and microsomal fractions and this procedure will prove of difinitive value in the diagnosis of the uterine cervix.
Cytological criteria of breast cancer for the purpose of mass screening is studied. Method of cytological examination must be aspiration biopsy smear, because the positive ratio of nipple discharge to diagnose the cancer is not so adequate. Staining of preparation have been done by means of May-Giemsa method. State of cell cluster, size of cells, color of cytoplasm, position of nuclei, shape of nuclei, size of nuclei, state of nuclear border and chromatin, size and numbers of nucleoli etc. have been observed about each cell of 100 cells of one preparation in the cases with carcinoma, mastopathy and fibroadenoma. 30 cases of diseases, above mentioned, were studied. 1) Over 20 μ of nuclear diameter means positive cytology. If largest nucleus in one preparation is under 10 μ, it means negative cytology. 2) Over 5μ of nucleolar diameter means positive, and over 5 nucleoli are observed, it means also positive. 3) Few cell between the cell clusters of smear preparation, it means negative. Many cells are scattered between the small clusters of preparation, it may be diagnosed as positive. 4) The largest nuclear diameter shows 2 times as the smallest one in one cell cluster, it means positive cytology, Degenerative nuclei for example pyknoticor karyolytic, have to be distinguished from the cells which keep the distinct chromatin structure. 5) Hyperchromatic nuclei in the cluster are smaller as nuclei which keep the clear chromatin structure, it means negative cytology. 6) The typical structure of the breast cancer cells obtained by aspiration is as follows. Nuclear border is not clear, chromatin is very fine and distributed evenly in the nucleus, and nucleolus has sometimes intranucleolar vacuoles.
1. The vaginal smear specimens of 113 patients with cervical cancer received radiation treatment were examined, and Cancer cell Decreasing Rate (CDR), Cancer cell Decreasing Curve (CDC), Radiation Response (RR) and Sensitization Response (SR) were calculated. 2. On nonmalignant squamous cells following changes were observed after irradiation: enlargement of nucleus and cytoplasm, vacuolation of nucleus and cytoplasm, pyknosis, karyorrhexis, multinucleation, cytolysis and abnormal staining. Among these changes enlargement was most prevalent. Also these changes were observed mor frequently in deep layer cells than in superficial layer cells. 3. These radiation changes, i.e. enlargement, vacuolation, multinucleation, thickened nuclear membrane, pyknosis, karyorrhexis and abnormal staining were also observed on cancer cells. Farthermore, the malignant cells rapidly disappeared in smear specimen on proceeding of the radiation treatment. 4. The CDR agreed quite well with CDC except for 2 cases, but showed no correlation with clinical stage of the patients nor developing type of the tumor. Poor CDR was more frequent in adenocarcinoma than in squamous cell carcinoma. 5. The patients with good CDR or good RR showed statistically better prognosis than those with poor CDR or poor RR respectively, but there was no difference between prognosis of the patients with good SR and those with poor SR. When RR and CDR agreed in a given patient, more accurate estimation of prognosis was attained
The influence of carcinostatic agents upon squamous cell carcinoma of extremities was studied in six cases cytologically and histologically. Out of them, 3 were thermal burn scar carcinoma. Mitomycin C (MMC) and Bleomycin (BLM) were used as carcinostatic agents. Three of four cases to whom MMC was administered underwent a local perfusion. The remaining two cases were given 15 mg of BLM intravenously three times weekly. The cytological changes of cancer cells by carcinostatic agents were summarized as follows 1) vacuole formation in cytoplasm and nucleus 2) multinuclear cell 3) enlargement of cell and nucleus 4) disarrangement of nuclear chromatin pattern 5) diminution of staining intensity. The prominent destruction of carcinoma pearl and the diminution of cancer cells accompanying with nuclear pyknosis or karyorrhexis were histologically observed in cancerous lesion. Moreover, nuclear pyknosis and decrease of basal cells were found in the normal basal layer. In conclusion, these changes of cancer cells are a nonspecific degeneration which is influenced by carcinostatic agents, but occurrs very frequently in noadministered cases. The changes of cancer cells shows only retrogressive degeneration. So appearance of degenerated cells is in parallel with the effect of carcinostatic agents, that the pursuit of cell changes may be of great usefulness in defining the influence of carcinostatic agents.
Cytological studies on vaginal smears of the newborn with special reference to relations between steroid metabolism and cytological changes. Fractionation was made of steroids in the urine of the new born; day to day determinations were performed of estriol and 17 OS in reference to cytological findings. 1) Predominantly abundant esteriol was determined on a day to day basis, revealing the highest amount 1-2 days after birth followed by a rapid decrease till no determination was made on the seventh postnatal day. On cytological vaginal smears there were observed the estrogen effect and a slight progesterone effect on the day of birth, a marked estrogen effect only a day after birth, and on estrogen effect five days after birth showing good correspondence with the results of determination. 2) In the fractination of 17 OS DHEA proved to be dominant. 17 OS showed no marked changes for five days after birth. It showed on increase five days after birth reaching a maximum on the seventh postnatal day. On cytological vaginal smears the androgen effect which was observed about seven days after birth showed good correspondence with the determined value of 17 OS. The development of the androgen effect at this stage seemed to be due to the disappearance of estrogen. 3) On cytological vaginal smears, it was assumed that seven days after birth the value of 17 OS showed an increase which was maintained till about 11 days after birth, followed by a decrease which lasted for same time after the 15 th postnatal day. 4) It was assumed that DHEA in 17 OS played an important role in the androgen effect.
The cytologic examination of bile was investigated to establish it as a definitive diagnosis of cancers of the gallbladder, the biliary tract and the liver. On the procedure of fixation, it was found that the fixative of 99.5% ethylalcohol was best to make cellular characteristics clear in the cytologic examination of bile. The abnormal cells in bile, which was aspirated from the gallbladder and the biliary tract during percutaneous transhepatic cholangiography, external biliary drainage and laparotomy, were demonstrated on the smear-slides with Papanicolaou's stain and the cell-block slides with H.E. stain. Data on the cytologic examination of bile of 74 patients was analyzed. The disease in seven cases was diagnosed from cytology of the bile. These cases included two carcinomas of the gallbladder, two carcinomas of the common bile duct, one hepatoma and two cases of metastatic carcinoma to the liver. No false positive observations were made. Of nine cases with dysplastic report, five cases were found to have one carcinoma of the gallbladder, one carcinoma of the common bile duct, three metastatic carcinomas to the liver diagnosed by histologic section, but the remaining four cases with dysplastic report were proved by the operative findings and the histologic diagnoses to be inflammatory diseases and gallstones. The missed cases, in which pathologic proof of cancer existed, included one carcinoma of the common bile duct and one hepatoma. In summary, the cytologic diagnosis of bile can be reliable for the definitive diagnosis of cancers of the gallbladder, the biliary tract and the liver.
In our uterine cancer clinic in the section of gynecology, Kyoto 1st Red Cross Hospital, we have been making particular efforts to detect uterine cancer at the early stage, by means of a systematic examination including vaginal smear test, colposcopy, and punch biopsy. We statistically summarized the results of the examination of uterine cancer on the patients who visited our clinic during recent 2 years (1968, -1969). Particular attention was paid to the results by the cytological diagnosis. The conclusion are summarized as follows; 1) The subject examined in this study was mainly a woman who was not younger than 30 years old. Total number of the patients was 3268, whose 71% was occupied by the women aged from 30 years old to 49 years old. 2) Concerning the subjective symptom when the patient visited our clinic to receive cancer examination, abnormal genital bleeding was first ranked complain at the frequency of 28% of the total cases. While 19% of the patients received cancer examination without any particular subjective symptoms. It should be promoted in future that more frequent number of women willingly receive the cancer examination before complaining any subjective symptoms. 3) Cytological diagnosis were fiven to 3268 cases, whose 92.5% (3025 individuals) were negative in cyto-diagnosis, whils 3% (98 individuals) were suspected positive and another 4.5% (145 individuals) were positive in cyto-diagnosis. 4) Histological examination was also applied to 1130 patients had received cytological diagnosis. The results proved that a reliability of the cytological diagnosis with vaginal smear could be estimated at about 91% in accuracy. The accuracy of the cytodiagnosis in the patients who were diagnosed cancer after the systemic examination was estimated at 96.4%, while the one of patients who were free of cancer by the same examination was estimated at 98.3%. 5) Our cancer clinic found 166 cases of the uterine cancer (including 10 cases of dysplasia) during this period of examination. The items of an account were; carcinoma in situ-7 cases, Iastage-36 cases, Ib stage-31 cases, II stage-78 cases, III stage-11 cases and IV stage-2 cases. 6) The ratio of detection of uterine cancer summarized as 87.5% in carcinoma in situ, 94.4 % in Ia stage, 100% in Ib stage, 97.5% in II stage, 90.9% in III stage and 100% in IV stage, (dysplasia was detected by cytological diagnosis at the ratio of 80%). 7) For clinical diagnosis, the cervical-erosion, especially the one of relatively slight degree was proved in most of the cases of uterine cancer at early stage. 8) The colposcopy was applied to the patients (1249 women), who were suspected uterine cancer by the cytological diagnosis or were found cervical-erosion by the gynecological examination. The uterine cancer was detected by colposcopy at the level of 89% in accuracy, thus proving lower reliability as compared to the cyto-diagnosis.