Pulmonary Venous Malformation in a 4-Year-Old Boy: a Case Report

We report a case of a pulmonary venous malformation in a 4-year-old boy who presented with recurrent pneumonia. A radiograph revealed a right infrahilar mass and a hyperlucent right lung. Computed tomography (CT) demonstrated a mass containing intensely enhancing areas and multiple phleboliths located in the right lower lobe and encasing the right bronchus and right inferior pulmonary vein. Magnetic resonance imaging (MRI) precisely revealed the mass demarcation. A right lower lobectomy was performed and a pathological examination confirmed the diagnosis of a venous malformation. To the best of our knowledge, a venous malformation in pulmonary tissue has not been reported in the English literature. Herein, we report a case of a pulmonary venous malformation, with the radiograph, CT, MRI, and blood pool scan findings, along with its pathologic correlation.

Expression and Localization of Epidermal Growth Factor in the Testosterone administered Submandibular Gland of the Rat / 대한해부학회지

The submandibular gland of rodents is under-developed state at birth, and the differentiation takes place post-natally under the control of neuronal and hormonal factors. In particular, testosterone plays an important role in the differentiation of duct system of rodent submandibular gland, but the underlying mechanism is not clear. To clarify the relationship between testosterone administration and differentiation of granular convoluted tubular (GCT) cells in immature rats, the expression and localization of epidermal growth factor (EGF), as a marker of GCT cell, on the submandibular gland of 3 week -old rats were examined by means of RT -PCR, immunohistochemistry and in situ hybridization. 1) RT-PCR demonstrated gradual increases of the amounts of EGF mRNA following the administration of testosterone for 1~4 days. 2) Immunohistochemistry shows that the occurrence of EGF protein first appeared at the striated duct after the administration of testosterone for 1 day, and had increased in number at the duct portions, 2 and 4 days after the testosterone administration. 3) In situ hybridization also indicates testosterone-induced expression of EGF mRNA became evident as EGF protein at the duct system of rat submandibular gland. These results suggest that testosterone is involved in the differentiation of the granular convoluted tubular cells, during the post-natal development of the rat submandibular gland.

Effect of Acriflavine-Guanosine Compound (AG60) Treatment on the Gastrointestinal Endocrine Cells of Mouse Inoculated with Ehrlich Carcinoma Cells / 대한해부학회지

To study the tumor-suppression effect of a newly developed anti-tumor agent AG60 [ acriflavine (1) : guanosine (1) composition, Taerim Pharm. Co., Seoul, Korea], each Ehrlich carcinoma (107 cells)-inoculated mouse received the subcutaneous injection of 0.2 ml of saline, 5mg/kg of AG60, and 30 mg/kg of AG60, every other day for two weeks. Animals were sacrificed, and stomach, duodenum, appendix vermiformis and rectal tissues were resected and fixed in 10% neutral formalin. Tissue blocks were washed, dehydrated, embedded and cut in 6 microgram-thick sections. For immunocytochemistry, the streptavidine-biotin-peroxidse method was used with a InnoGenex (San Ramon, Calif., USA) staining kit. The tissues were incubated with rabbit antisera against somatostatin (Biogenesis, Poole, England, UK) diluted 1 : 300, secretin (Biogenesis, Poole, England, UK) diluted 1 : 2,400, neurotensin (Biogenesis, Poole, England, UK) diluted 1 : 2,600, or motilin (Biogenesis, Poole, England, UK) diluted 1 : 1,000 for 24 hour at 4dreeges C, followed by incubation in biotinylated antirabbit IgG and horseradish peroxidase-streptavidin conjugate for 1 hour at room temperature. The antigen-antibody reaction sites were visualized by incubating the sections with diaminobezidine tetrahydrochloride (DAB) for 5~15 minutes at room temperature. After mounting in canada balsam, they were examined in a Leica DM RB microscope. The number of the immunoreactive cells in the area of gastrointestinal mucosae (mean number of immunoreactive cells per 0.25mm2) were observed and calculated. The results are as follows : 1. In the fundic gland of normal mouse, somatostatin immunoreactive cells were detected (18.5+/-0.71), but neurotensin, secretin, or motilin immunoreactive cells were not found. In the duodenal mucosa of normal mouse, somatostatin immunoreactive cells were detected (7.0+/-0.10), but neurotensin, secretin or motilin immunoreactive cells were rarely found. 2. Immunoreactivity of somatostatin, secretin, neurotensin or motilin cells was not found in appendix vermiformis and rectum of normal mouse. 3. On immunocytochemical study, somatostatin immunoreactive cells in the fundic glands of normal, experimental control, AG60 (5mg/kg)-treated, AG60 (30 mg/kg)-treated and 5-fluorouracil (60 mg/kg)-treated groups were 18.5+/-0.71, 10.0+/-4.20, 11.5+/-0.71, 13.5+/-2.10, 11.5+/-2.71, respectively. 4. On immunocytochemical study, somatostatin immunoreactive cells in the duodenal mucosae of normal, experimental control, AG60 (5 mg/kg)-treated, AG60 (30 mg/kg)-treated and 5-fluorouracil (60 mg/kg)-treated groups were 7.0+/-2.10, 0.5+/-2.71, 3.0+/-1.41, 0.5+/-0.71, 2.50+/-0.71, respectively. 5. On immunocytochemical study, secretin immunoreactive cells in the duodenal mucosae of normal, experimental control, AG60 (5 mg/kg)-treated, AG60 (30 mg/kg)-treated and 5-fluorouracil (60 mg/kg)-treated groups were rarely found. 6. On immunocytochemical study, neurotensin and motilin immunoreactive cells in the duodenal mucosae of normal groups were detected, but immunoreactivies were not detected in experimental control, AG60 (5 mg/kg)-treated, AG60 (30 mg/kg)-treated or 5-fluorouracil (60 mg/kg)-treated groups.

Morphological Study on the Thymic Cortex of the Ehrlich Carcinoma-Inoculated Mice, Treated with 5-Fluorouracil and Acriflavine-Guanosine Complex (AG60) / 대한해부학회지

In cancer therapy, immunological disorder is one of most severe problem. Since thymic cortex is the home of T-cell proliferation and "education", thymic morphology following administration of certain drugs can be used as a parameter of immunological safety of the drug. In this study, morphology of thymic cortex, following administration of 5-fluorouracil or AG60, was studied. AG60 is a newly developed anti-cancer remedy, the compound of acriflavine and guanosine (1 : 1). ICR mice were subcutaneously inoculated with Ehrlich carcinoma cells (10(7) cells/mouse) in their inguinal areas. Each mouse in 5-fluorouracil group was injected subcutaneously with a single dose of 30 mg/kg of 5-fluorouracil every other day, and the mouse in AG60 group, with 30 mg/kg of AG60 (Taerim Pharm. Co., Seoul) every other day. The control mouse was injected with saline. The mice were sacrificed on the day after 7th injection. Tissues of thymic cortices were fixed in 2.5% glutaraldehyde-1.5% paraformaldehyde solution (0.1 M Millonig's phosphate buffer, pH 7.3), and refixed in 2% osmium tetroxide solution (0.1 M Millonig's phosphate buffer, pH 7.3). Tissue blocks were dehydrated, and were embedded in araldite mixture. For the overview-comparison, semithin sections stained with toluidine blue solution were photographed. And the typical portions were cut with ultratome, stained and observed with electron microscope. In light microscopy, thymic cortical morphology of AG60-injected mouse was similar with that of control mouse. But the cortical morphology of 5-fluorouracil-injected mouse was impressively different from those of the control or AG60 group mice. Thymocytes in the thymic cortex of 5-fluorouracil-injected mice were severely depleted. In electron microscopy, thymocytes in the thymic cortices of the control or AG60 group mice were crowded, and small groups of thymocytes were surrounded by the cytoplasmic processes of epithelial reticular cells. Mitotic figures were randomly seen. Thymocytes of 5-fluorouracil-injected mouse were naked out from the epithelial reticular cells, and were completely depleted out from the cortex composed mainly of enlarged epithelial reticular cells. Numerous microvilli were protruded from the naked thymocytes. The results were interpreted as that 5-fluorouracil induce leukopenia, and homing of lymphocytes to thymic cortex is severely depressed. 5-fluorouracil also disturb the normal protective and supportive function of epithelial reticular cells for thymocytes. Whereas the complex of acriflavine-guanosine compound (AG60) is immunologically safe, as seen in thymic cortical morphology.

Effects of Adriamycin or AG60 to the DNA Synthesis of Duodenal Epithelium of Mice Implanted with Ehrlich Carcinoma Cells: An Autoradiographic Study / 체질인류학회지

This experiment was performed to evaluate the morphological responses of the intestinal gland of the mouse, duodenum inoculated with Ehrlich carcinoma cells, following administration of adriamycin or acriflavine -guanosine composition (AG60, Taerim Pharm. Co. Seoul, Korea). Healthy adult ICR mice weighing 25 g each were divided into normal and experimental groups (experimental control group, adriamycin treated group, and AG60 treated group). In the experimental groups, each mouse was inoculated with 1 x10 7 Ehrlich carcinoma cells subcutaneously in the inguinal area. From next day, 0.2 ml of saline, adriamycin (2 mg/ kg), AG60 (5 mg/kg) or AG60 (30 mg/kg) were injected subcutaneously to the animals every other day, respectively. The day following the 7th injection of anticancer drugs, each mouse was injected with a single dose of 0.7 microCi/gm of methyl -3 H -thymidine (25 Ci/mmol, Amersham Lab., England) through tail vein. Seventy minutes after the thymidine injection, animals were sacrificed. The number of the labeled epithelial cells of the duodenal crypts (mean number of labeled epithelial cells per 3.5 mm length of mucosa) were observed and calculated. On histological study, in the duodenum of adriamycin treated groups, vesiculated epithelial cells of the intestinal villi, expanded lumen of the intestinal gland (G) and loosely arranged lamina propria were observed. But in the AG60 treated group, morphological changes of the duodenum were not observed. On autoradiographic study, number of the labeled cells of normal control, experimental control, adriamycin -treated, AG60 (5 mg/kg)-, and AG60 (30 mg/kg)-treated groups were 595.3 +/-48.96, 715.+/-89.11, 96.0 +/-15.62, 632.0 +/-83.16 and 370.3 +/-49.65, respectively. In the adriamycin and AG60 30mg/kg -treated group, poorly -labeled cells containing only a few silver grains of 3 H -thymidine were observed more frequently than in those of the normal control group. But in the experimental control group, number of the heavy labeled cells were observed more frequently than in those of the normal control group. From the above results, adriamycin and AG60 (30 mg/kg) may suppress the DNA synthesis of the cells of the duodenal crypts. But AG60 does not result any histological defect on the duodenal mucosa. These results suggest that AG60 is expected as one of most effective anticancer drugs.

Effects of Adriamycin or CP -2 on the Spleen of Mouse Implanted with Ehrlich Carcinoma Cells: An Autoradiographic Study / 체질인류학회지

In this experiment, side effects of two anticancer drugs (adriamycin and CP -2) on the structure of spleen were histologically studied. Each of ICR mice was inoculated with 1 x10 7 Ehrlich carcinoma cells subcutaneously in the inguinal area. From next day, 0.2 ml of saline solution, adriamycin (2 mg/kg) or CP -2 (30 mg/kg) were injected subcutaneously every other day. The day following the 7th injection of adriamycin or CP -2, each mouse was injected with a single dose of 0.7 micro Ci/gm of methyl -3 H -thymidine (25 Ci/mmol, Amersham Lab., England) through tail vein. Seventy minutes after the thymidine injection, animals were sacrificed, and splenic tissues were collected and fixed in 10% neutral formalin. Deparaffinized sections were coated with autoradiographic emulsion EM -1 (Amersham Lab., England) in the dark room and dried, and were kept in a light -tight box. The sections were exposured for 5 weeks in the dark room, and were developed in D -19 developer. The number of the labeled cells in the areas of the white pulp, the red pulp and the marginal zone (mean number of labeled cells per 0.21 mm 2 ) were observed and calculated. In the spleen of adriamycin treated group, vacuoles containing pyknotic nuclei were observed frequently. Whereas in the CP -2 treated group, morphological changes of the spleen were not observed. The number of the labeled cells of normal control, experimental control, CP -2 treated and adriamycin treated groups were 240.3 +/-53.28, 252.3+/- 58.24, 216.7 +/-55.17 and 45.4 +/-15.46, respectively, and most of the labeled cells were located near the marginal zone of the spleen. In the adriamycin treated group, labeled cells containing a few silver grains of 3 H -thymidine were observed more frequently than in those of the normal and experimental control groups. From the above results, adriamycin and CP -2 may suppress the DNA synthesis of the splenic tissues. Especially, CP -2 does not results any histological defect on the splenic tissues. These result suggest that CP -2 is expected as one of effective anticancer drugs.

Ultrastructural Alterations Induced by 5-Fluorouracil or Mitomycin C on the Gastric Parietal Cells of Mouse / 대한해부학회지

This experiment was performed to evaluate the morphological responses of 5-fluorouracil or mitomycin C on the gastric parietal cells of mouse. 5 -fluorouracil (30 mg/kg) or mitomycin C (400 micro gram/kg) were injected subcutaneously every other day, and the animals were sacrificed at 4th day and 7th day following the first injection. Pieces of the tissue were taken from the stomach, prefixed with 2.5% glutaraldehyde -1.5% paraformaldehyde, followed by post-fixation with 1% osmium tetroxide. The ultrathin sections were stained with uranyl acetate and lead citrate. In both of the 5-fluorouracil or the mitomycin C treated groups, most parietal cells showed severely reduced luminal spaces of the intracellular canaliculi, since microvilli of intracellular canaliculi were very irregular shaped and nearly contacted with each other, and the cytoplasmic tubulovesicular membranes were disintegrated and indistinct. The changes in the 5-fluorouracil treated group were more indistinct than in those of the mitomycin C treated group. In the 5-fluorouracil treated group, balooning of the cytoplasm, focal cytolysis, myelin figures, lysosomes and multivesicular bodies in the parietal cells were observed more frequently than in those of the mitomycin C treated group. Above results suggest that the 5-fluorouracil or mitomycin C treated animals might suffer from reduced acid secretion of the parietal cell, since the collapsed lumen of the intracellular canaliculi, the disintegration of the tubulovesicular membranes, and the reduction of cell organelles in the parietal cells are occurred within a few days following injections. 5-fluorouracil was proved more harmful on the parietal cell than mitomycin C does.

Morphological Study on the Mouse Spleen following the Administration of Mercuric Chloride or Potassium Dichromate / 대한해부학회지

This experiment was performed to study the morphological changes of the spleen of mice following injection of sodium dichromate (K2Cr2O7) or mercuric chloride (HgCl2). Male mice were divided into normal and experimental groups. The mice were subcutaneously injected with mercuric chloride (5 mg or 10 mg/kg) or sodium dichromate (10 mg or 20 mg/kg). Animals were sacrificed on 6 hours, 1 day, 3 days, 1 week and 2 weeks after injections. Pieces of splenic tissue were taken from each mouse, and fixed in 10% neutral formalin for light microscopy. The paraffin sections were stained with hematoxylin-eosin, Masson-trichrome, Bielschowsky's silver impregnation or aldehyde-fuchsin stain. For electron microscopy, the tissues were fixed in 2.5% glutaraldehyde-1.5% paraformalde-hyde, and post-fixed in 1% osmium tetroxide. Dehydrated blocks were embedded in araldite mixture. The ultrathin sections stained with uranyl acetate and lead citrate were observed with JEM 100CX-II electron microscope. On histological study, in the early stage (6 hours) of experimental groups, splenic white pulp exhibited numerous vacuoles containing pyknotic nuclei were observed as compared with those of normal control group. But after 3 days(sodium dichromate, 10 mg/kg or 20 mg/kg; mercuric chloride, 5 mg/kg) and 1 week (mercuric chloride, 10mg/kg), the morphology was recovered to normal one. In the experimental groups, positive reactions to Bielschowsky's silver impregnation, Masson-trichrome or aldehyde-fuchsin stain were similar to those of normal control group. On the ultrastructural study, in white pulps of experimental groups, nuclear bodies were observed frequently in the nuclei of the lymphocytes and the reticular cells, and myelin figures were observed in the nucleus or in the cytoplasm of the lymphocytes and the reticular cells. The plasma cells showed many irregularly distended cisternae of granular endoplasmic reticula and the macrophages containing phagosomes, were observed frequently. From the above results, it was concluded that potassium dichromate or mercuric chloride could disturb the normal differentiation or maturation of the lymphocytes and the reticular cells of the spleen, especially in the early stage of treatment. But histological changes occurred in the spleen following injection of the potassium dichromate or mercuric chloride were recovered to normal appearance in 3 days (potassium dichromate) or 1 week (mercuric chloride). Mercuric chloride was more harmful than potassium dichromate on the spleen.

Thymic Neuroendocrine Tumor Associated with Cushing's Syndrome: case report / 대한흉부외과학회지

Neuroendocrine tumor in thymus is rare and has poor prognosis due to frequent recurrence and distant metastasis. Approximately half of thymic carcinoids are hormonally active and Cushing's syndrome is seen in 33% of affected patients. Treatment of choice is surgical excision of tumor and role of chemotherapy and radiotherapy is controversal. We report 2 cases of thymic neuroendocrine carcinoma associated with Cushing's syndrome.

Effects of Bacillus Calmette-Guerin (BCG) on the Spleen of Mouse Inoculated with Ehrlich Carcinoma Cells: A Morphological Study / 대한해부학회지

This experiment was performed to study the morphological responses of the spleen of mouse inoculated with Ehrlich carcinoma cells, following administration of Bacillus Calmette-Guerin (BCG). Healthy adult ICR mice weighing 25 g each were divided into normal and experimental groups (experimental control group and BCG treated group). In the experimental groups, each mouse was inoculated with 1X10(7) Ehrlich carcinoma cells subcutaneously in the inguinal area. From next day, 0.2 ml of saline or BCG (0.03X10(8)-0.32X10(8) CFU) were injected subcutaneously to the animals every other day. The day following the 7th injection, each mouse was injected with a single dose of 0.7 microCi/gm of methyl-3H-thymidine (25 Ci/mmol, Amersham Lab., England) through tail vein. Seventy minutes after the 3H-thymidine injection, animals were sacrificed. Pieces of the splenic tissue, fixed in 10% neutral formalin for light microscopy. The sections were coated with autoradiographic emulsion EM-1 (Amersham Lab., England) and the coated sections were exposured for 5 weeks in the dark room. For electron microscopy, tissues were prefixed with phosphate buffered 2,5% glutaraldehyde-1.5% paraformaldehyde (pH 7.3), and post-fixed with phosphate buffered 1% osmium tetroxide solution (pH 7.3). Ultrathin sections of the white pulp area stained with uranyl acetate and lead citrate were observed with a JEM 100CX-II electron microscope. On histological study in the splenic white pulp, BCG treated mice showed more macrophages containing pyknotic nuclei than normal or experimental control mice showed. On autoradiographic study, a large number of the 3Hthymidine labeled cells were seen near the marginal zone, whereas only a small number of labeled cells were seen in the red pulp or the white pulp of the spleen. The number of the labeled cells in experimental control group was similar to that in the normal control mice, whereas that in BCG-treated mice was significantly increased as compared with that of normal control one. On electron microscopic study, in the white pulp of BCG treated mouse, mitotic cells were observed more frequently than in those of the normal or experimental control mice. In the BCG treated mice, macrophages and plasma cells in the white pulp were observed more frequently than in those of the normal or experimental control mice, whereas a few eosinopile leucocytes were observed, and perichromatin granules within the nuclei of the lymphocytes and the reticular cells were observed frequently. From the above results, it was concluded that DNA syntheses were more active in the cells of the marginal zone than in the cells of the white pulp or the red pulp. And repeated treatment with BCG could activate the DNA syntheses of splenic cells and increase the number of the macrophages and the plasma cells in the white pulp.

Effects of Antitumor Agents on Ultrastructure of the Splenic White Pulp of the Mouse Implanted with Ehrlich Carcinoma Cells / 대한해부학회지

This experiment was performed to study the morphological responses of the splenic white pulp, the lymphatic tissue of the spleen, of Ehrlich carcinoma cell-implanted mice to three different anticancer drugs (5-fluorouracil, mitomycin C and AG60). Healthy adult ICR mice weighing 20 g each were divided into normal and experimental groups. In the experimental groups, each of mice was inoculated with 1X10(7) Ehrlich carcinoma cells subcutaneously in the inguinal area. From next day, 0.2 ml of saline solution, 5-fluorouracil (30 mg/kg), mitomycin C (400 microgram/kg) or AG60 (30 mg/kg, Taerim Pharm. Co., Seoul, Korea) were injected subcutaneously every other day, and animals were sacrificed at 14th day following the f irst injection. Pieces of the tissues were taken from the spleen, and prefixed with phosphate buffered 2.5% paraformaldehyde-1.5% glutaraldehyde solution (pH 7.3) followed by post-fixation with phosphate buffered 1% osmium tetroxide solution (pH 7.3). Fixed tissue blocks were dehydrated, and embedded in araldite mixture. Ultrathin sections stained with uranyl acetate and lead citrate were observed with a JEM 100CX-II electron microscope. In the experimental control group (carcinoma cell-inoculated mouse), splenic white pulp did not show pronounced morphological alterations, but myelin figures were frequently observed in the cytoplasm of some lymphocytes and reticular cells than those of normal control mice. In the AG60 treated group, splenic white pulp did not show specific morphological defect, but nuclear bodies and severe invaginations of the nuclear envelope of the lymphocytes and reticular cells were observed occasionally. In the mitomycin C treated group, myelin figures, severe invaginations of the nuclear envelope, nuclear protrusions, nuclear bodies and interchromatin granules were frequently observed in the lymphocytes and reticular cells of the white pulp. In the 5-f luorouracil treated group, myelin f igures, severe invaginations of the nuclear envelope, nuclear protrusions, nuclear bodies and interchromatin granules were observed more frequently in the lymphocytes and reticular cells of the white pulp, as compared with those of mitomycin C treated mice. From the above results, 5-f luorouracil or mitomycin C may suppress the splenic immune function of cancerinoculated mice, since they suppress the process of differentiation and maturation of splenic lymphocyte and reticular cells, and 5-fluorouracil was more harmful on the spleen than mitomycin C. Whereas AG60 does not affect remarkably the process of differentiation and maturation of lymphocytes and reticular cells in the splenic white pulp.

Effects of Lower Dose-Injections of Acriflavine-Guanosine Compound (AG60) on the DNA Synthesis and Ultrastructure of the Ehrlich Carcinoma Cells Inoculated to the Mouse / 대한해부학회지

AG60, a recently introduced anti-cancer compound, was reported to show highly effective anti-cancer activities, when injected with doses from 30 mg/kg to 5 mg/kg.The purpose of this study was to know the lower effective doses of AG60, and to give the informations for preparing more advanced therapeutic tools for anti-cancer war. Ehrlich cancer cells were inoculated in the subcutaneous tissue of inguinal region of ICR mice, and saline (treated control groups) or AG60 (experimental groups) were injected daily. Animals of experimental groups were injected subcutaneously with doses of 0.2 mg/kg, 0.5 mg/kg, 1.0 mg/kg, or 2.0 mg/kg body weight, according to their subgroups. Five mice from each subgroup were sacrificed on 1 week, 2 weeks, 3 weeks following the first injection. Seventy minutes before sacrifice, each mouse was injected with 0.7 microCi/g body weight of 3H-thymidine (Amersham Lab.) through tail vein. After sacrifice, cancer masses were fixed in 10% formalin solution for autoradiography and light microscopy, and in 2.5% glutaraldehyde-1.5% paraformaldehyde solution, followed by post-fixation in 2% osmium tetroxide solution, for electron microscopy. The observed results were as follows: Autoradiographic observations show, 1. Labelled cancer cell indices of the experimental groups received AG60 were decreased around to 80% (0.2 mg/kg), to 74% (0.5 mg/kg), to 75~60% (1.0 mg/kg), and to 70~50% (2.0 mg/kg), as compared with those of the controls. 2. The contents of silver grains were dramatically decreased nearly to 35% (0.2 mg/kg), to 20% (0.5 mg/kg), to 21~16% (1.0 mg/kg), and to 20~15% (2.0 mg/kg), as compared with those of the controls. 3. Total granular content in 100 cancer cells on the third week of the experiment decreased nearly to 30% (0.2 mg/kg), to 15% (0.5 mg/kg), to 10% (1.0 mg/kg), and to 8% (2.0 mg/kg), as compared with those of the controls. Histological observations show, 1. AG60 induces large amount of apoptosis on Ehrlich cancer cells. 2. Following the treatment with AG60, multinuclear cells or giant cells were increased in number. Comparing by autoradiography and histology, multinuclear or giant cells were interpreted as those cells supplied by poor amounts of thymidine, or almost no new DNA content. Electronmicroscopic readings show, 1. AG60 induces numerous macroclefts and microclefts within the nuclei of Ehrlich cancer cells. 2. AG60 induces numerous apoptosis among Ehrlich cancer cells. 3. Apoptotic bodies are phagocytosed by adjacent cancer cells or by macrophages. From the above results, AG60 is expected to be a very successful anti-cancer candidate. And it is suggested that combined or cocktail therapy including AG60 may greatly improve the anti-cancer therapy on certain kind of cancer.

Morphological Changes of Cultured Rat Hepatocytes Induced by Phalloidin and Cytochalasin D / 대한해부학회지

The microfilaments of hepatocyte are distributed throughout the vicinity of cell membranes, especially numerous around the region of bile canaliculus, and provide the maintenance of cell shape, cellular wall tension, canalicular motility, the secretion for bile, etc. To evaluate the relationship between the microfilament and alteration of cell shape, we examined the morphological changes of cultured rat hepatocytes, following treatments with phalloidin or cytochalasin D with fluorescent and electron microscopes. 1. In the fluorescent micrographs, actin microfilament was distributed near the plasma membrane and bile canaliculus. 2. Both drugs, phalloidin or cytochalasin D, produce the cytoplasmic protrusions from the surface. Their shapes were pedunculated with narrow neck or bulged with broad base, respectively. 3. In the phalloidin treated group, cytoplasmic protrusion was seperated from the internal cytoplasm by microfila-ments networks at the narrow base. In contrast, in the cytochalasin D treated group, cytoplasm was bulged with broad base and kept in direct continuity with the canalicular ectoplasm. 4. Pericanalicular ectoplasm of phalloidin treated group was widened and accumulated with microfilaments. But, bile canaliculus of cytochalasin D treated group was markedly dilated and devoid of microvilli, and the ectoplasm was almost disappeared. Considering above results, dysfunction of microfilaments leads to the structural changes and inhibition of bile secretion of hepatocytes.

Age Related Morphological Study on the Spleen of Mouse / 대한해부학회지

In this study, the structural components of mouse spleen were compared during their aging processes. Splenic tissues of 1 week-, 5 weeks-, 8 weeks-, 6 months-, 12 months-, 18 months-, 24 months-, and 30 months-old ICR mice were dissected out under anesthesia. Pieces of the tissues were taken from the spleen, fixed in 10% neutral formalin for light microscopy, and some splenic tissues, were prefixed with 2.5% glutaraldehyde-1.5% paraformaldehyde followed by post-fixation with 1% osmium tetroxide for electron microscopy. Paraffin sections stained with hematoxylin-eosin, Masson's trichrome, Bielschowsky's silver impregnation or aldehyde-fuchsin, were observed with light microscope. Ultrathin sections stained with uranyl acetate and lead citrate were observed with a JEM 100CX-II electron microscope. The observed results were as follow: 1. The thickness of the splenic capsule was prominently increased between 1 week-old and 5 weeks-old ones, whereas after 5 weeks-old, the thickness was very slightly increased during aging. 2. In the 1 week-old, 5 weeks-old and 8 weeks-old mice, blood forming cells were observed more frequently than those found in older ones. 3. The collagenous fibers and elastic fibers were increased in the spleens of 12 months-old mice, whereas after 18 months-old, fibers were not increased during aging. 4. The reticular fibers were increased by 8 weeks, whereas fibers were not increased afterwards. 5. In the 1-week old, mast cells were observed frequently, whereas from 5 weeks to 6 months they were observed rarely, and after 12 months, mast cells were observed frequently. 6. In the 5-weeks old, distended cisternae of granular endoplasmic reticula were first observed within the plasma cells. 7. After 12 months, the mast cell containing phagocytosed debris were observed frequently. 8. After 18 months, the plasma cell containing irregular distended cisternae of granular endoplasmic reticula were observed frequently. 9. In 30 months, the plasma cells containing myelin figures were observed frequently. From the above results, it was suggested that spleen of the mouse matures structurally in five weeks, but the function of the spleen is suppressed around 18 months, and thereafter the functional suppression is continued on aging.

Effects of Antitumor Agents on the Spleen of Mouse Implanted with Ehrlich Carcinoma Cells: An Autoradiographic Study / 대한해부학회지

In this experiment, side effects of three anticancer drugs (5-fluorouracil, mitomycin C and AG60) on the structure of spleen were histologically studied. Each of ICR mice was inoculated with 1X10(7) Ehrlich carcinoma cells subcutaneously in the inguinal area. From next day, 0.2 ml of saline solution, 5-fluorouracil (30 mg/kg), mitomycin C (400 micorgram/kg), 5 mg/kg of AG60, or 30 mg/kg of AG60 (acriflavine (1) : guanosine (1) composition, Taerim Pharm. Co., Seoul, Korea) were injected subcutaneously every other day. The day following the 7th injection of anticancer drugs, each mouse was injected with a single dose of 0.7 microCi/gm of methyl-(3)H-thymidine (25 Ci/mmol, Amersham Lab., England) through tail vein. Seventy minutes after the thymidine injection, animals were sacrificed, and splenic tissues were collected and fixed in 10% neutral formalin. Tissue blocks were washed, dehydrated, embedded and cut into 6 micrometer-thick sections. Deparaffinized sections were coated with autoradiographic emulsion EM-1 (Amersham Lab., England) in the dark room and dried, and were kept in a light-tight box. The sections were exposured for 5 weeks in the dark room, and were deveoped in D-19 developer. The number of the labeled cells in the areas of the white pulp, the red pulp and the marginal zone (mean number of labeled cells per 0.21 mm2) were observed and calculated. The results were as follow: 1. On histological study, in the spleen of mitomycin C treated group, vacuoles containing pyknotic nuclei were observed more frequently than in those of 5-fluorouracil treated group. Whereas in the AG60 treated group, morphological changes of the spleen were not observed. 2. On autoradiographic study, most of the labeled cells were located near the marginal zone of the spleen. 3. On autoradiographic study, number of the labeled cells of normal control, experimental control, 5-fluorouracil-treated, mitomycin C-treated, AG60 (5 mg/kg)-, and AG60 (30 mg/kg)-treated groups were 214.0+/-56.87, 235.7+/-59.69, 331.0+/-67.20, 137.0+/-33.48, 124.6+/-34.28, and 64.9+/-16.26, respectively. 4. In the mitomycin C treated group and AG60 (5 mg/kg or 30 mg/kg) treated group, labeled cells containing a few silver grains of (3)H-thymidine were observed more frequently than in those of the normal and experimental control groups. But in the 5-fluorouracil treated group, number of the heavy labeled cells were observed more frequently than in those of the normal and experimental control groups. From the above results, DNA synthesis, in the cells of the marginal zone were more active than in the cells of the white pulp or the red pulp. And mitomycin C and AG60 may suppress the DNA synthesis of the splenic tissues. Especially, AG60 does not results any histological defect on the splenic tissues. These result suggest that AG60 is expected as one of most effective anticancer drugs.

A Case of Retroperitoneal Cystic Lymphangioma

Cystic lymphangiomas are comparatively rare, benign tumors of lymphatic system and their histogenesis are uncertain. About 75% of theses lesions are in the neck, 20% are in the axillary region, and 5% are in the mediastinum, retroperitoneal region or groin. But retroperitoneal cystic lymphangiomas are very rare. Retroperitoneal cystic lymphangiomas are usually found incidentally during diagnostic procedures performed for unrelated clinical reasons or at surgery. Although retroperitoneal cystic lymphangioma is a benign lesion, it may cause significant morbidity due to its large size and its often invasive character with a strong tedency to secondary infection. They generally present as a palpable mass or abdominal pain and fever related to hemorrhage or inflammation of the cystic wall. The treatment of choice is surgical excision. We experienced a case of retroperitoneal cystic lymphangioma in a 4-year-old boy who had developed rapid abdominal distention and abdominal pain for 4 days. We have completely excised large cystic mass & histologically confirmed cystic lymphangioma. A brief review of literature was made.

Morphological Study on the Spleen following the Administration of 5-Fluorouracil or Mitomycin C / 체질인류학회지

This experiment was performed to evaluate the morphological responses of 5-fluorouracil or mitomycin C on the spleen of the mice. 5-fluorouracil (60 mg/kg) or mitomycin C (400 microgram/kg) were injected subcutaneously every other day, and the animals were sacrificed at 1 week and 2 weeks following the first injection. Pieces of the tissue were taken from the spleen, fixed in 10% neutral formalin for light microscopy. The paraffin sections stained with hematoxylin-eosin, Masson-trichrome, Bielschowsky's silver impregnation or aldehydefuchsin. For electron microscopy, the tissues were prefixed with 2.5% glutaraldehyde-1.5% paraformaldehyde followed by post-fixed with 1% osmium tetroxide. The ultrathin sections stained with uranyl acetate and lead citrate. The observed results were as follows: 1. On histological study, in the mitomycin C treated group, macrophages which contain pyknotic nuclei were observed more frequently than in those of 5-fluorouracil treated group. 2. In the 5-fluorouracil treated group, positive reactions to Masson-trichrome and Bielschowsky's silver impregnation were observed in the splenic capsule and traculae at the 1 week, and weak postive stains were observed at the 2 weeks. 3. In the mitomycin C and the 5-fluorouracil group, positive staining reaction to aldehyde-fuchsin were observed in splenic capsule, trabeculae and around artery at the 1 week and 2 weeks. 4. On the ultrastructural study, distended cisternae of granular endoplasmic reticula were observed frequently at 1 week. 5. In the mitomycin C treated group, myelin figures in the lymphocytes and reticular cells were observed more frequently than in those of 5-fluorouracil treated group. From the above results, it was concluded that lymphocytes and reticular cells of the spleen were slightly damaged by 5-fluorouracil or mitomycin C, and mitomycin C seems more harmful on the spleen than 5-fluorouracil does.

Ultrastructural Changes of the Cultured Hepatocytes in Microfilamentous Dysfunction induced by Drugs / 대한해부학회지

To examine the role of actin microfilaments which are located at beneath the plasma membrane, we observed the ultrastructural changes of rat hepatocyte induced by alteration of the microfilamentous integrity. The isolated hepatocytes from Sprague-Dawley were cultured in the L-15 medium containing phalloidin (agent that cause polymerization of actin) or cytochalasin D (agent that cause depolymerization of actin) for 30 min, 1 hour, 2 hours, 4 hours, 10 hours and 20 hours, respectively. The results observed with scanning and transmission electron microscope were as follows. 1. Following the alteration of actin microfilaments, bile canaliculi were dilated and devoid of microvilli. In phalloidin treated group, the thickening of microfilamentous ectoplasm was more marked than that of cytochalasin D treated group. Whereas, the dilation of bile canaliculi was more marked in cytochalasin D group. 2. Both drugs, phalloidin or cytochalasin D, produced the alteration of cell shape to form cytoplasmic protrusions at the cell surface. In the phalloidin treated group, protrusions were pedunculated, and the microfilament networks were accumulated at the narrow neck region. 3. In cytochalasin D treated group, no microfilament barrier was seen at the broad base of protrusion which exhibit direct continuity with the internal cytoplasm. 4. Single hepatocyte tend to recover their structural integrity as those in vivo. The new bile canaliculus was sealed off at the intercellular space by tight junctions, and intercellular contacts were established by the junctional complexes. The results demonstrated that excessive accumulation or depletion of microfilaments induced by phalloidin or cytochalasin D altered the cell shape different, respectively. The microfilaments of ectoplasm play an important role in the maintenance of the structural integrity of cultured hepatocytes.

Fluorescent Study on the Tissue-Distribution of Acriflavine-Guanosine Compound (AG60) / 대한해부학회지

The study was carried out to evaluate the tissue-distribution of acriflavine or AG60 (acriflavine-guanosine compound, 1 : 1), the newly developed anticancer remedy. Successful access or distribution of a drug to specific tissue is important to attack the cancer cells in the same area. But it also means that the drug may disturb the activities of labelled tissues or cells. On the other hand, unlabelled elements may survive from massive treatment with the drug. In this study, distribution of acriflavine or AG60 in Yac-1 leukemic cells (0.25~25 microgram/ml) and in the tissues of rats or mice (5~50 mg/kg) were evaluated. Yac-1 cells showed prominent fluorescence on the heterochromatin and more or less prominent fluorescence on the nucleoplasm, cytoplasm and plasma membrane. Cytotoxicity of AG60 led to morphologic changes such as bleb- or baloon-formation on the surface, general swelling of the cell, and lysis of the cell. Following the subcutaneous administration of acriflavine or AG60 (5~50 mg/kg) to the Ehrlich carcinoma-inoculat-ed rats or mice, most tissues including cancer cells showed acriflavine-fluorescence with some exception. The nuclei of cells of tissues were labelled more strongly than those of cytoplasm. Fluorescence was especially strong over biliary tree, renal corpuscle, gastrointestinal mucous coat, and bronchial mucous coat. But parenchymal components of central nervous system did not show any fluorescence. As shown in Yac-1 cells treated with AG60, the drug strongly attached to nucleic acids, and it induced swelling and disintegration of cancer cells. Fast turn-over of AG60 was seen in the secretory passages of bile juice, urine, gastrointestinal mucin, and bronchial mucin. The results show that AG60 could reach most tissues except parenchymes of central nervous system.

Ultrastructural Changes of the Ventricular Choroid Plexus of the Rat following Irradiation on the Head / 대한해부학회지

Acute irradiation effects on the choroid plexus were studied. Healthy male rats (Sprague Dawley) were anesthetized with sodium thiopental, and were placed on the table of Mistubishi linear accelerator ML-4MV. The rats were irradiated on their heads and necks with the doses of 3,000 rads or 6,000 rads. Six hours, 2 days and 6 days following the irradiations, rats were sacrificed by perfusion fixation through the heart, with 1% glutaraldehyde-1% paraformalde-hyde solution. Choroid plexuses of lateral ventricles were taken out and fixed in the same fixative overnight, and were postfixed in 2% osmium tetroxide solution. Fixed tissue blocks were dehydrated within series of alcohol and acetone, and were embedded in araldite mixture. Ultrathin sections were cut, and contrasted with uranyl acetate and lead citrate solutions, and observed with electron microscope. The results obstained were as follows : 1. Choroid epithelial cells of irradiated rats exhibited severe alterations of their organelles concerning intracellular material transport; such as disappearance of microvilli and basal infoldings, reduction of invaginating pits on the basal and apical plasma membranes, reduction of transfer vesicles, shrinked Golgi complexes, etc. 2. In choroidal epithelial cells of the rats exposed with doses of 6,000 rads, the morphological alterations of the intracellular transport system were more severe than those of the rats exposed with doses of 3,000 rads. 3. Cellular elements and collagen fibrils within the pericapillary space were not so much affected following irradiations. 4. Zonula occludentes in between choroidal epithelials cells were not altered after the irradiations. The above results indicate that the production of cerebrospinal fluid by choroid plexus is severely reduced. Since the choroid plexus functions not only the secretions but it also concerns the homeostasis of many important ions, neuromodulaters, and other peptides, heavy irradiation on the head may result severe malfunction of central nervous system and humoral homeostasis.