The Incidence Rate and Severity of Orthotopic Lung Cancer in an Animal Model Depends on the Number of A549 Cells and Transplantation Period / 한국실험동물학회지

The incidence rate of lung cancer is continually increasing, and lung cancer is the leading cause of cancer-related death worldwide. Nevertheless, few therapeutic methods are available for lung cancer. Therefore, establishing appropriate lung cancer animal models is important to investigate mechanisms and to evaluate new drugs for lung cancer. In the present study, we transplanted non-small cell lung cancer A549 human adenocarcinoma cells (2x10(4), 2.0x10(5), and 2.0x10(6) cells) into the right lobe of BALB/c nude mice via the intercostal space to develop an orthotopic lung cancer animal model that is minimally invasive and similar to human lung cancer. We then investigated the incidence rate and severity of lung cancer according to the A549 cell number (2x10(4), 2.0x10(5), and 2.0x10(6) cells) and transplantation periods (4~23 days). Lung cancer development was confirmed with gross examination, which was supported by histopathological examination. These results indicate that the incidence rate and severity of lung cancer was increased depending on the number of transplanted cells and transplantation period which the cell number and duration are increasing risk of lung cancer. Thus, this study can provide appropriate reference data to develop an orthotopic lung cancer animal model using the non-small cell lung cancer A549 cell line for researching mechanisms and evaluating candidate drugs, including various approaches for treating lung cancer.

Point Mutation of Hoxd12 in Mice

PURPOSE: Genes of the HoxD cluster play a major role in vertebrate limb development, and changes that modify the Hoxd12 locus affect other genes also, suggesting that HoxD function is coordinated by a control mechanism involving multiple genes during limb morphogenesis. In this study, mutant phenotypes were produced by treatment of mice with a chemical mutagen, N-ethyl-N-nitrosourea (ENU). We analyzed mutant mice exhibiting the specific microdactyly phenotype and examined the genes affected. MATERIALS AND METHODS: We focused on phenotype characteristics including size, bone formation, and digit morphology of ENU-induced microdactyly mice. The expressions of several molecules were analyzed by genome-wide screening and quantitative real-time PCR to define the affected genes. RESULTS: We report on limb phenotypes of an ENU-induced A-to-C mutation in the Hoxd12 gene, resulting in alanine-to-serine conversion. Microdactyly mice exhibited growth defects in the zeugopod and autopod, shortening of digits, a missing tip of digit I, limb growth affected, and dramatic increases in the expressions of Fgf4 and Lmx1b. However, the expression level of Shh was not changed in Hoxd12 point mutated mice. CONCLUSION: These results suggest that point mutation rather than the entire deletion of Hoxd12, such as in knockout and transgenic mice, causes the abnormal limb phenotype in microdactyly mice. The precise nature of the spectrum of differences requires further investigation.

Status of parasitic infection diagnosed by surgical biopsy in Kwangju and Chollanam-do

: In order to know the species and frequency of human parasitic infection diagnosed by biopsy, 149 cases (0.18%) of parasitic infection were reviwed, which were selected from 80,947 biopsied materials submitted for routine histopathological examination during a period of 10 years from 1980 to 1989 at Department of Pathology, Chonnam National University Hospital. They consisted of 112 cases of cysticercosis, 17 paragonimiasis, 7 clonorchiasis, 4 amebiasis, 1 sparganosis, 1 enterbiasis, 1 aniskiasis, and 1 fascioliasis respectively. Based on morphological preservation of cysticercus, they could be divided into mild (20.2%), morderate (40.4%), and severe (39.4%) degeneration. Except 2 cases biopsied at the lungs, 15 cases of ectopic paragonimiasis were located at abdominal cavity (8 cases) and central nervous system (7 cases). One case of intrahepatic fascioliasis was observed. This is the 13th human fascioliasis reported in Korea. From the above results, the frequency of parasitic infections found in biopsied specimens was on the decrease as the cysticercosis and ectopic paragonimiasis.

Histological, Enzyme Histochemical, and Electron Microscopic studies of the Rat Kidney Following Administration of Folic Acid

To investigate the morphologic evidence of acute renal failure by folate, histological, histochemical (PAS), enzyme histochemical (Na-K-ATPase, G6PD, and ALP), and ultrastructural studies were performed. The results are as follows: l) Oliguria was most severe 3 hours after folate and the urine volume was 24.8% that of the control group. 2) Histologically, dilatation of tubules, degeneration and focal necrosis of the cortical tubules, and PAS(+) droplets in the tubular lumen were noted. And also frequent mitoses, mild interstital connective tissue proliferation, and neutrophilic infiltrates were observed in the late stage. 3) On enzyme histochemical examination, the activities of Na-K-ATPase and ALP were decreased, but G6PD activity was increased in comparison with the control group. 4) The ultrastructural studies revealed cytoplasmic vacuoles, apical cytoplasmic blebbing, dense bodies, mildly swollen mitochondria, dilated endoplasmic reticulum, loss bf brush border of the proximal tubules, and loss of microvilli of the thin limb of Henle's loop. Later, marked attenuation or loss of infoldings of basal plasma membrane of the cortical tubules was recognized. According to above results, the cause of acute renal failure by late is thought to be the injuries of tubular epithelial cells including sodium pump secondary to tubular obstruction.

An Image Analytical Study on the Structural Spectrum of Intestinal Metaplasia-Dysplasia-Carcinoma of the Stomach

Intestinal metaplasia and dysplasia of the stomach have been stressed as precursors of gastric carcinoma of the intestinal type, although their preneoplastic nature is still debated. In this study, the cytomorphometric and cytokinetic spectra of the suggested preneoplastic and neoplastic lesions of the stomach were investigated. From the resected stomachs of early gastric carcinoma of intestinal type, areas of normal, intestinal metaplasia, dysplasia, and carcinoma were selected. They were immunostained for proliferating cell nuclear antigen, counterstained with propidium iodide, and various nuclear parameters were measured by image analysis. Normal and intestinal metaplastic mucosae differed by the localization of proliferation zone, but not by nuclear profile area, circular shape factor, and proliferation index. In dysplasia, proliferation zone covered large parts of the dysplastic area. Nuclear profile area and proliferation index were larger whereas circular shape factor was smaller than in normal or intestinal metaplasia. Carcinomatous lesion had diffuse proliferation activity, the largest nuclear profile area and proliferating index, and circular shape factor in-between those of normal or intestinal metaplasia and dysplasia. The above results showed a structural spectrum among normal of intestinal metaplasia, dysplasia, and carcinoma of intestinal type in cytomorphometric and cytokinetic terms. The structural spectrum raises the possibility that dysplasia of the stomach is a preneoplastic lesion.

DNA ploidy and Cellular Proliferation Activity in Experimentally Induced Malignant Fibrous Histiocytoma

To fine out the changes of DNA ploidy and cellular proliferation activity during carcinogenesis and evaluate correlation between flow cytometrically determined S-phase fraction and proportion of proliferation cell nuclear antigen(PCNA, PC10) immunoreactive cells, the authors studied on malignant fibrous histocytoma induced by intra-articular injection of 9, 10-dimethy1-1, 2-benzanthracene(DMBA) in the rats. Forty Wistar rats were used. The results obtained were as follows. 1) Firstly, tumors were palpated 5 weeks after the last injection of DMBA and formed in 27 rats at sacrificed. Histologically, these lesions showed storiform, indicative of malignant fibrous histiocytoma. 2) Three cases of DNA aneuploidy were observed at 4 and 5 months after the last injection of DBMA and one of them, which was DNA diploidy at main mass, was found at daughter mass. 3) Flow cytometrically determined S-phase fraction and proportion of PCNA(PC10) immunoreactive cells in malignant fibrous histiocytoma induced by DMBA were much higher than in control groups and slightly increased according to sequential changes after formation of mass. The comparison of flow cytometrically determined S-phase fraction and proportion of PCNA(PC10) immunoreactive cells showed significant correlation(r=0.6092, p<0.001). Above results strongly suggest that ploidy pattern may evolve into aneuploid type during the development of tumor and proliferation activity increases during the carcinogenesis.

Correlation between Expression of c-erbB-2 Oncogene and Various Prognostic Factors in the Colorectal Carcinoma

The c-erbB-2 oncogene, which is a new human proto-oncogene similar to EGFR structurally, generates a glycoprotein of tyrosine kinase family with a molecular weight of 185,000 To evaluate the prognostic significance of c-erbB-2 oncogene expression in colorectal carcinoma, We analysed 73 colorectal carcinomas in paraffin sections immunohistochemically, using the monoclonal antibody specific for the c-erbB-2 oncogene product and correlated with clinicopathological data. The results were as follows 1) The immunoreactivity for c-erbB-2 oncogene was localized to cell membrane of the tumor cells and occasionally observed within the cytoplasm. 2) The positivity of c-erbB-2 oncogene expression was 71.2%(52/73) of the colorectal carcinomas overall. According to the histological types, the positivity of c-erbB-2 oncogene in adenocarcinoma(77.4%) was higher than that in mucinous carcinoma(36.4%)(p<0.05). 3) Expression of c-erbB-2 oncogene was significantly correlated with lymph node metastasis or distant metastasis(p=0.0117), Dukes stage(p=0.0432), and TNM classification(p=0.0102). These results suggest that c-erbB-2 oncogene expression may be used as a prognostic factor of colorectal carcinoma because of its correlation with other clinicopathological prognostic factors.

Chronological Observation of an AIDS Patient from Onset to Death and Post-mortem Autopsy Study

The ‘32-year-old sailor’ patient was a native Korean male, infected with HIV through heterosexual contacts in the high risk areas of foreign country, mostly during. 1984 to 1986. During the following 6-months until his death, the skin nodules suggesting Kaposi's sarcoma, and penile ulcers like chancroid were gradually disappeared. Several skin diseases such as condyloma accuminatum, generalized xeiosis of the skin, acquired icthyosis, seborfheic dermatitis, diffuse hair loss and alopecia developed. His health gradually deteriorated with general symptom such as weakness, weight loss, remAtmt mild fever cough with sputum, and dyspnea with frequent pneumonia signs. The tongue became coated white and thick in which candida albicans was detected, and sore throat and dysphagia accompanied. The patients expired suddenly with acute respiratory distress syndrome, which was may be caused by a opportunistic infection of Pneumocystis carinii. In post-mortem autopsy studies, most of the lung tissue showed the findings of severe interstitial pneumonia and hyaline membrane formation, and pneumocystis cadnii and candida albicans were found in histopathological or micorbiological examination. As other infections, add fast bacilli suggesting Mycobacterium avium-intracellulare were found out in the lymph node, spleen, liver. Cytomegalovirus infection was found on the adrenal gland by histopathological examination. And various lymphoid tissue of the lymph node, spleen, thymus, respiratory tract, gastomtestinal tracts showed severe lymphoid depletion with fibrosis. The other findings had multiple glial nodules in the cerebral white matter and focal segmental glomerulosderosm There wasr t any evidence of Kaposi's sarcoma.

A Case of Angiodysplasia in the Jejunum / 대한소화기내시경학회지

Angiodysplasia of the small bowel is an uncommon disorder in Korea, and is often found to be the cause of unexplained gastrointestinal bleeding. A 53-year-old male patient visited an emergeney room because of repeated melena and dizziness, He underwent selective superior mesenteric arterial angiography which revealed abnormal clusters of small arteries and early opacification of veins draining a segment of lower jejunum. An exploratory laparotomy was performed under the impression of angiodysplasia. An ulceration on the lower jejunum was found, and was resected. Subsequent jejunojejunostomy was performed. Histology of the resected specimen showed an irregularly dilated vessels in mucosa and submucosa.

A Case of Angiodysplasia in the Jejunum / 대한소화기내시경학회지

Angiodysplasia of the small bowel is an uncommon disorder in Korea, and is often found to be the cause of unexplained gastrointestinal bleeding. A 53-year-old male patient visited an emergeney room because of repeated melena and dizziness, He underwent selective superior mesenteric arterial angiography which revealed abnormal clusters of small arteries and early opacification of veins draining a segment of lower jejunum. An exploratory laparotomy was performed under the impression of angiodysplasia. An ulceration on the lower jejunum was found, and was resected. Subsequent jejunojejunostomy was performed. Histology of the resected specimen showed an irregularly dilated vessels in mucosa and submucosa.

Immunohistochemical and Ultrastructural Cellular Differentiation in Papillary and Solid Epithelial Neoplasm of the Pancreas

Papillary and solid epithelial neoplasm of the pancreas from five patients were studied using immunohistochemistry and electron microscopy to define the cellular origin of this type of tumor. The tumors ranged in diameter form 5.5 to 15 cm Grossly, these were well circumscribed by a firm, gray-white, fibrous capsule and their cut-surface showed mainly area containing mucinous substance with necrotic and hemorrhagic material, with some solid portion. Microscopically, there was a solid and papillary pattern, with uniform cells typically having round to ovoid nuclei containing indistinct nucleoli and eosinophilic, granular cytoplasm. Within the cytoplasm of the tumor cells, numerous PAS-positive granules were found. Immunostaining was positive for neuron-specific enolase(three of five cases), alpha1-antitrypsin and alpha1-antichymotrypsin(three of five cases) in the solid and papillary portion of the tumor. But no polypeptide hormone immunoreactive cells were present in all cases except for gastrin which showed focally weak positivity in the papillary area. Ultrastructurally, the papillary and solid epithelial neopasm of the pancreas showed evidence of acinar cell differentiation, because in the cell of one observed some zymogen-like granules and presence of annulate lamellae. But also, abundant typical neurosecretory granules were detected in the tumor cells ultrastructurally. Both facts suggested acinar and islet cell differentiation of the tumor. From the these findings, it concluded that papillary and solid epithelial neoplasm of the pancreas may be originated from a primordial cell which will be able to render both endocrine and exocine component.

Cytopathologic Observation of Primary Malignant Melanoma of the Lung: A case report

The pulmonary cytology has reached a high level of accuracy. By the examination of the sputum and/or bronchial brushings, it is now possible to make a diagnosis in 70% to 90% of patients with cancer. Primary melanoma of the lung is very rare and there have been reported about 20 cases in the world literature. We present a case of primary malignant melanoma of the lung in a 61-year-old male diagnosed by cytologic examination of sputum, bronchial brushing and aspirated pleural fluid. Histologic examination of bronchoscopic biopsy and examination of the skin and other primary sites confirmed the diagnosis.

Evaluation of DNA Ploidy and Other Morphometric Parameters of Ovarian Mucinous Tumors

Biological behavior of malignant tumors has been assessed by morphological grading, clinical staging, and estimating other tumor markers. Recently DNA ploidy measured by flow cytometry and image analyser has been suggested as an additional useful indicator of the tumor behavior. In order to extract useful tumor cell-specific information in ovarian mucinous tumors, DNA contents and other morphologic parameters were measured by image analysis and DNA ploidy was also measured by flow cytometry. In all cases of cystadenoma, DNA diploidies were observed. In borderline malignancy, DNA diploidies were chiefly observed except one case of polyploidy. In true malignancy, DNA aneuploidies were observed except one case of polyploidy and two cases of diploidies by image analysis, and except four cases of diploides and one cas of polyploidy by flow cytometry. The statistical significance were observed in DNA ploidy pattern by image analysis. In nuclear areas, perimeters and major axis, statistical significance were not observed. These results suggest that DNA ploidy pattern are more or less independent parameter as an additional useful indicator of the histological grade of malignancy and that image analysis are better than flow cytometry in detecting DNA aneuploidy.

The Expression of ras Oncogene in Benign and Malignant Lesions of Breast

To evaluate correlation between the amount of oncogene products in tumor cell extracts and malignant potentiality in breast tumor, immunohistochemical staining for the ras Oncogene products was performed in the sections of benign and malignant lesions of the breast. The results obtained were as follows: 1) The positive reaction to ras Oncogene products was usually observed in the cytoplasm and cell membrane. 2) The ratio of positive reaction was 30.4% in epithelial hyperplasia of fibrocystic disease, 26.5% in fibroadenoma. 49.5% in intraductal carcinoma 71.6% in infiltrating ductal carcinoma, 85.2% in metastatic infiltrating ductal carcinoma, and 89.7% in relatively preserved neighboring lobules of infiltrating ductal carcinoma. In conclusion, the ras oncogene products are found by a significantly higher ratio in the more aggressive lesions, and the infiltrating ductal carcinoma might represent its potential of malignant transformation. 3) The expression of ras oncogene was heterogeneous in primary as well as metastatic mammary carcinomas.

A Study on the Cell Kinetics of the Dysplastic Epithelium in the Stomach

This study was designed to evaluate the biological behavior of the dysplastic lesion of the stomach by applying immunohistochemical method for bromodeoxyuridine (BrdUrd). The results obtained were as follows. 1) In most hyperplastic and dysplastic lesions, the proliferative cell zones, loci of BrdUrd-labelled cells, were found in the upper later of the mucosa, whereas they were confined to the neck zone in the normal gastric mucosa. 2) The labelling indices (LIs), percentages of BrdUrd-labelled cells, were 11.0% to 13.6% in the normal gastric mucosa, and were 14.3% to 17.9%, 16.4% to 19.2% and 17.4% to 20.7% in the simple hyperplasia, in the atypical hyperplasia and in the dysplasia, respectively. These findings suggested that proliferative potential in hyperplasia and dysplasia were greater than that in normal gastric mucosa, the higher the grade of dysplasia being, the greater the proliferative potentials.

An Evaluation of Results of Cytological Diagnosis for the Uterine Cervical Lesions

It awakens interest that the frequency of carcinoma of the uterine cervix is the highest among the carcinomas among the Korean women, althouth it is not so high among Westerns. It is admitted that the exfolicative cytologic technique is useful not only as a screening test for early detection of cervical cancer and its precursor lesions but also as a tool for follow-up, because it is accurate, economical, and easy to perform. But it remains to be solved that the false negativity of the cytologic diagnosis is relatively high. Five hundred and sevently two cases of cytology specimens, confirmed by either colposcopic biopsy or cone biopsy, were studied among 15,844 cases which were submitted to the Department of Pathology, Chonnam University Hospital between October of 1982 and August of 1986. The results obtained were as follows. 1) The accuracy of the cytologic diagnosis was 69.9%, the false nagativity, 19.4% and the false positivity, 6.6%. 2) Among the causes of discrepancies between cytologic diagnosis and histologic diagnosis, error in cytology reading was the most frequent (57%), error in cell collection was the second frequent (43.7%), and error in biopsy location was the least frequent (5.3%). 3) Among the 38 cases of false positive diagnosis, the percentage of error in cytology reading was 82.3%, and that of error in biopsy location was 20.7%. Among the 105 cases of false negative diagnosis, the percentage of error in cytology reading was 50.5%, and the percentage of error in cell collection was 56.6%.

Congenital Melanocytic Schwannoma in Ankle Joint Potentially Malignant: A case report

Congenital malignant melanocytic schwannoma in ankle joint was not reported on literature and it was a very interesting case. Light microscopically, melaninladen cells were mixed in abundant wavy spindle cells, some mitotic cells were also observed. Ultrastructurally, melanosomes in variable stages of development were scattered in the cytoplasm which had basal lamina. Collagen bundles were abundant in the intercellular connective tissue. It was histologically malignant tumor and clinically recurred. But in non-congenital potentially malignant melanocytic schwannoma which had been reported, reccurrence or distant metastasis were not noted. So, further clinical survey may be necessary for evaluation of the malignant behavior of this neoplasm.

The Expression of ras and myc Oncogene in Transitional Cell Carcinoma of the Urinary Bladder

The oncogenes, which have been detected in various human solid tumors, transform culture cells, and the level of m-RNA specific for an oncogene increases in the cellular extract of the tumor cells. These findings suggested that oncogene expression was closely related with carcinogenesis. Recently, oncogen products were considered as tumor markers, but it was not confirmed that the relationship between quantitative change of oncogene product and malignant potential of a neoplasm. To evaluate the relationship between the quantitative change of oncogene product and malignant potential, immunohistochemical staining for the ras and myc oncogene products was performed in the sections of papilloma and transitional cell carcinoma of urinary bladder. 1) Positive reaction of c-ras oncogene product was noted along the cell membrane and in the cytoplasm, and c-myc oncogene product in the nucleus, and along the unclear membrane and cell membrane. 2) Tissue expression of c-ras oncogene was homogeneous and strong in the transitional cell carcinoma rather than in papilloma. 3) The ratio of the positive cells with c-ras oncogene product was 35.1% in the papilloma, 79.4% in the grade I, 81.9% in the grade II, 87.6%, in the grade III of transitional cell carcinoma of the urinary bladder. There was a tendency for the ratio to increase with the degree of histological grading. 4) By the immunoperoxidase staining of c-myc oncogene product, the number of the cells showing positive nuclear staining incrased with the tumor grading.

Light and Electron Microscopical Studies on the Stroma of Hydatidiform Mole

Many investigators were interested in the pathogenesis and the relationship between microscopical features and clinical behavior of hydatidiform mole. Trophoblastic cells in the trophoblastic disease were intensively examined histologically, ultrastructurally, immunohistochemically, and with hormone assay method, etc. But ultrastructural study on the stroma of hydatidiform mole was scarcely reported. In this paper, hydatidiform mole was examined at light and electron microscopic levels, with emphasis on the stroma. The results were as follows: 1) Hydropic degeneration of H-mole is more severe in the center of stroma and is not related with the degree of trophoblastic proliferation. Hofbauer cell and vascular structure are extremely rarely observed in the periphery of stroma which has relatively preserved cellular components. 2) Basement membrane is sometimes separated from trophoblastic layer. Degenerated cells in the stroma contain vacuoles, autophagosomes, and lipid droplets. Collagen is abundant in the loose interstitium. Hofbauer cells have no lysosome or phagosome. Vascular lumen is patient and endothelial cells are degenerated. From the above results, H-mole may be produced due to abnormal changes of trophoblasts and stromal changes may be a secondary process, so called autolysis. Hofbauer cells are not engaged in the stromal degeneration and may be different from usual tissue macrophages.

Morphologic Study on the Mesothelial Hyperplasia in the Rat

Reactive mesothelial cells are frequently found in the smear, of body fludid dffusion when associated with liver cirrhosis, pulmonary tuberculosis, congestive heart failure or pulmonary infarction. But it is not easy to differentiate these reactive mesothelial cells from macrophages or various types of malignant cells. In order to investigate the morphology of hyperplastic mesothelial cells, in comparison with the various types of malignant cells in the effusion, this experiment was designed. The tissue reaction was evoked by intraperitoneal injection of 2.5 ml of 2% Talcum suspension. The results obtained were as follows: 1) On light microscopic observation of the tissure, until 5 days, cytoplasm and nucleus of mesothelial cells became larger and nucleoi became prominent. After then, cytoplasm and nucleus became smaller and at 28 days after injection of Talcum suspension, cellular features became similar to those of normal ones. 2) On cytologic observation of touch print specimens, hyperplastic mesothelial cells became round to oval, and their cytoplasm became conspicuous. Nuclei were vesicular, and nucleoi were prominent. N/C ratio was decreased than normal. 3) On electron microscopic observation, hyperplastic mesothelial cells did not have microvilli, but had a few cytoplasmic globular projections, loosely approximated cellular junctions, larger and more abundant intracytoplaomic vesicles when compared with normal one. Chromatin clumping, irregular thickening of nuclear membrane and prominent nucleoi were also noted.