Morphological Study of Respiratory Portion of Mouse Lung following Chemical Glue-Vapor Inhalation / 대한해부학회지

Industrial glues contain many kinds of organic solvents and glue sniffing by young people has become a social problem in Korea. Glue vapor may induce chronic toxicities different from those induced by exposures to the solvent of single component. We studied the effects of the inhalation of glue vapor on the primary target organ, the pulmonary epithelium of the respiratory system. Vapor samples of glue were collected for analysis; the components were acetone, n-hexane, methyl cyclopentane, c-hexane, and toluene. For the inhalation of glue vapor, experimental mice were exposed in a whole body chamber for 20 min/d for 3, 5, 7, and 14 d. Control groups were exposed to room air. Animals were euthanized and lung tissues were fixed in 10% neutral formalin for light microscopy, and in 2.5% glutaraldehyde plus 1.5% paraformaldehyde for electron microscopy. The results are as follows. 1. Alcianophilic bands were not detected in the normal alveolar epithelium, but weak alcianophilic bands were detected in bronchioles. Alcian blue-PAS and PAS positive cells were found in the mucosae of mice exposed to glue vapor for 5 and 7 d. 2. Types I and II pneumocytes and capillary endothelial cells were found in the normal alveolar epithelium. The blood-air barrier consists of Type I pneumocytes, a common basal lamina, and the capillary endothelium. 3. The alveolar epithelium of vapor-exposed mice showed more type II pneumocytes. In the longerexposed group, Type I pneumocytes and endothelial cells contained many pinocytotic vesicles. 4. The vapor-exposed lungs showed macrophages in the alveolar space, mild interstitial swelling, and increased numbers of collagenous fibers. Clearly, ultrastructural changes in pulmonary epithelia can occur following glue sniffing.

Study on the Cerebral Laterality Pattern of Medical Students / 체질인류학회지

In this study, cerebral functional laterality patterns of medical students in the year 2005 were compared with those in the year 1995. Questionnaires on the behavior patterns were asked, and the laterality patterns were classified as right hemispheric (R)-balanced hemispheric (B)-left hemispheric (L). 385 students were studied (210 male and 175 female). Of the 3 categories, male students showed the patterns of R (42.8%), B (31.9%) and L (25.3%). Female students showed R (45.2%), B (30.9%) and L (23.9%). As the above result shows, laterality patterns of male and female were similar. The above data were compared with the another data in the article reported in 1995. Previous report showed that R (63.5%), B (24.9%) and L (11.6%) in the male students, and R (49.0%), B (22.4%) and L (28.6%) in female students, respectively. From the above results, It was suggested that cerebral laterality patterns of medical students in the year 2005 shifted toward left, but it still remained 42~45% in right hemispheric as contrast to 24~25% in left hemispheric. Hemispheric shift was interpreted as that, it may be the result of student's adaptative or competitive activities in the fast changing social environment.

Effects of Antitumor Agents to the DNA Synthesis of Cecal Mucosa of Mouse / 체질인류학회지

This experiment was performed to evaluate the morphological responses of the cecal mucosa of the mouse, inoculated with Ehrlich carcinoma cells in the inguinal area, following administration of 5-fluorouracil, mitomycin C or adriamycin. Healthy adult ICR mice weighing 25 gm each were divided into normal and experimental groups. In the experimental groups, each mouse was inoculated with 1 x 10(7) Ehrlich carcinoma cells subcutaneous in the inguinal area. From next day, 0.2 mL of saline, 5-fluorouracil (30 mg/kg), mitomycin C (400 microgram/kg) or adriamycin (2 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 micro Ci/gm of methyl-3H-thymidine (25Ci/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 cecal crypts (mean number of labeled epithelial cells per 3.5 mm length of mucosa) were observed and evaluated. On histological study, in the experimental control and mitomycin C-treated mice, general morphology of the cecal mucosae was similar. And in the 5-fluorouracil-treated mice, slightly swelled epithelial cells and expanded lumen of the intestinal crypts were observed. But in the adriamycin-treated groups, slightly disrupted intestinal crypts, a large number of basophilic epithelial cells and the expanded lumen of the intestinal crypts were observed. On autoradiographic study, number of the labeled cells of normal control, experimental control, 5-fluorouracil treated, mitomycin C-treated, or adriamycin-treated groups were 362.2+/-56.12, 350.7+/-71.13, 215.7+/-80.55, 144.2+/-34.60 and 125.0+/-37.45, respectively. In the adriamycin and mitomycin C-treated groups, poorly-labeled cells containing only a few silver grains were observed more frequently than in those of the normal and experimental control groups. From the above results, adriamycin and mitomycin C suppressed the DNA synthesis of the epithelial cells of the cecal mucosa more severely as compared with 5-fluorouracil did. Especially, adriamycin was more harmful than mitomycin C and 5-fluorouracil on the cecal mucosae.

Effects of BCG or CP-2 on the DNA Synthesis in the Epithelial Cells of the Mouse Appendix / 대한해부학회지

This experiment was performed to evaluate the morphological responses of the appendicular mucosa of the mouse, inoculated with Ehrlich carcinoma cells in the inguinal area, following administration of BCG or CP-2 (Coptis chinensis-Croton tiglium extracts). Healthy adult ICR mice weighing 25 gm each were divided into normal and experimental groups (experimental control, BCG or CP-2 treated group). Each experimental group mouse was inoculated with 1 x 10(7) Ehrlich carcinoma cells subcutaneously in the inguinal area. From the next day after inoculations, 0.2 mL of saline, BCG (0.5 mL/25 g B.W.: 0.03 x 10(8) ~ 0.32 x 10(8) CFU) or CP-2 (30 mg/kg) were injected subcutaneously to the animals every other day, respectively. The day following the 7 th injection of BCG or CP-2, 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 tritiated thymidine injection, animals were sacrificed. The number of the labeled epithelial cells of the appendicular mucosae were observed and evaluated. On histological studies of the experimental control, BCG or CP-2 treated mice, general morphologies of the appendicular mucosae were similar. On autoradiographic study, number of the labeled cells of normal control, experimental control, BCG treated or CP-2 treated groups were 362.2+/-56.12, 350.7+/-42.65, 265.8+/-27.08 and 241.3+/-53.29, respectively. Above results show that BCG and CP-2 suppress the DNA synthetic activity of the epithelial cells of the appendix, but did not show any remarkable morphological alterations on the mucosae. These results suggest that BCG and CP-2 are ones of effective anticancer drugs for the cytostatic therapy.

Effects of Acriflavine-Guanosine Composition (AG60) on the DNA Synthesis and Ultrastructure of Epithelial Cells of the Appendix of Mice Inoculated with Ehrlich Carcinoma Cells / 대한해부학회지

This experiment was performed to evaluate the morphological responses of the mucosa of the mouse appendix, inoculated with Ehrlich carcinoma cells in the inguinal area, following administration of Acriflavine-Guanosine Composition (AG60). Healthy adult ICR mice weighing 25 gm each were divided into normal, experimental control and AG60 treated group. Experimental control and AG60 treated groups, mice were subcutaneously inoculated with 1 x 10(7) Ehrlich carcinoma cells in the inguinal area. From next day after the carcinoma cell inoculations, 0.2 mL of saline or AG60 (5 mg/kg/0.2 mL) were injected subcutaneously to the animals every other day, respectively. The day following the 7 th injection of saline or AG60, 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, and appendix tissues were fixed in 10% formalin solution for light microscopy. The number of the labeled mucosal epithelial cells of the appendix were observed and evaluated. For the electron microscopic study, the tissues were fixed in 2.5% glutaraldehyde-1.5% paraformaldehyde solution, followed by post-fixation with 1% osmium tetroxide solution. Ultrathin sections were counter stained with uranyl acetate-lead citrate solutions, and observed. On light microscopic observation of experimental control and AG60 treated mice, did not show any remarkable morphological alterations on the mucosae. On autoradiographic study, number of the labeled cells within 3.5 mm width mucosae of normal control, experimental control, AG60 treated mice were 362.2+/-56.12, 350.7+/-42.65 and 90.7+/-33.48, respectively. On ultrastructural observation of the experimental control and AG60 treated mice, general morphologies of the epithelial cells of appendix were similar. But intranuclear filamentous structures, intramitochondrial dense granules, and myelin figures were occasionally observed in the absorptive cells of AG60 treated mice than control ones. Above results show that AG60 suppress the DNA synthetic activity of the mucosal epithelial cells of mouse appendix, but did show slight ultrastructural alterations in the absortive cells. These results suggest that AG60 is one of effective anticancer drug for the cytostatic therapy.

Effects of Antitumor Agents to DNA Synthesis in the Gastric Epithelia of Mice Implanted with Ehrlich Carcinoma Cells: An Autoradiographic Study / 대한해부학회지

This experiment was performed to evaluate the morphological responses of the gastric epithelium of the mouse, inoculated with Ehrlich carcinoma cells, following administration of 5-fluorouracil, adriamycin or mitomycin C. Healthy adult ICR mice weighing 25 gm each were divided into normal control and experimental groups (tumor control group, 5-fluorouracil treated group, adriamycin treated group, and mitomycin C treated group). In the experimental groups, each mouse was inoculated with 1 x 10(7) Ehrlich carcinoma cells subcutaneously in the inguinal area. From next day after inoculations, 0.2 mL of saline, 5-fluorouracil (30 mg/kg), adriamycin (2 mg/kg) or mitomycin C (400 microgram/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-3H-thymidine (25Ci/mmol, Amersham Lab., England) through tail vein. Seventy minutes after the thymidine injection, animals were sacrificed. The number of labeled epithelial cells in the gastric mucosae (mean number of labeled epithelial cells per 3.5 mm length of mucosa) were observed and calculated. On histological study, in the gastric mucosae of adriamycin-treated groups, denatured surface epithelial cells, expanded lumen of the gastric gland, and congested lamina propria were observed. But in the 5-fluorouracil or mitomycin treated groups, severe morphological changes of the gastric mucosae were not observed. On autoradiographic study, numbers of the labeled cells in the gastric mucosae per 3.5 mm length of normal control, tumor control, 5-fluorouracil-treated, adriamycin-treated and mitomycin C treated groups were 267.3 (+/-48.86), 273.6 (+/-59.41), 375.3 (+/-83.36), 15.3 (+/-9.66) and 124.0 (+/-32.66), respectively. In the adriamycin and mitomycin C-treated group, poorly-labeled cells containing only a few silver grains of 3H-thymidine were observed more frequently as compared in those of the normal control group. But in the 5-fluorouracil-treated 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 mitomycin C may severely suppress the DNA synthesis of the epithelial cells of the gastric mucosae. But some amount of the 5-fluorouracil (30 mg/kg) may not suppress the DNA synthesis of gastric epithelial cells.

Effects of Adriamycin or CP-2 to the Rectal Mucosae of Mouse Implanted with Ehrlich Carcinoma Cells: An Autoradiographic Study / 대한해부학회지

This experiment was performed to evaluate the morphological responses of the rectal intestinal glands of the mouse, inoculated with Ehrlich carcinoma cells, following administration of adriamycin or composition of the extracts of the Croton tiglium and Coptis chinensis rhizome (CP-2, Institute of Experimental Tumor Research, Seoul, Korea). Healthy adult ICR mice weighing 25 gm each were divided into normal and experimental groups (tumor control group, adriamycin treated group, and CP-2 treated group). In the experimental groups, each mouse was inoculated with 1 x 10(7) Ehrlich carcinoma cells subcutaneously in the inguinal area. From next day, 0.2 mL of saline, adriamycin (2 mg/kg) or CP-2 (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- 3H-thymidine through tail vein. Seventy minutes after the thymidine injection, animals were sacrificed. and rectal 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. The number of the labeled epithelial cells of the rectal crypts (mean number of labeled epithelial cells per 3.5 mm length of mucosa) were observed and calculated. On histological study, in the rectum of adriamycin treated groups, length of the intestinal crypts is shorter than those of the normal control ones. Disrupted intestinal crypts and epithelial cells were observed. But in the CP-2 treated group, morphological changes of the rectum were not observed. On autoradiographic study, number of the labeled cells of normal control, rumor control, adriamycin-treated, CP-2-treated groups were 263.1 (+/-38.65), 395.7 (+/-52.52), 73.3 (+/-22.54), 96.3 (+/-28.36), respectively. In the adriamycin and CP-2 treated groups., poorly-labeled cells containing only a few silver grains of 3H-thymidine were observed more frequently than in those of the normal and tumor control groups. But in the tumor 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 CP-2 may suppress the DNA synthesis of the cells of the rectal crypts. But CP-2 does not result any histological defect on the rectal mucosa. These results suggest that CP-2 is expected as one of most effective anticancer drugs.

Effects of 5-Fluorouracil, Mitomycin C or AG60 to the DNA Synthesis of Rectal Epithelium of Mice Implanted with Ehrlich Carcinoma Cells / 체질인류학회지

This experiment was performed to evaluate the morphological responses of the intestinal gland of the mouse, rectum inoculated with Ehrlich carcinoma cells, following administration of 5- fluorouracil, mitomycin C or AG60. Healthy adult ICR mice weighing 25 gm each were divided into normal and experimental groups (tumor control group, 5-fluorouracil, mitomycin C treated group, and AG60 treated group). In the experimental groups, each mouse was inoculated with 1*10 (7) Ehrlich carcinoma cells subcutaneously in the inguinal area. From next day, 0.2 mL of saline, 5-fluorouracil (30 mg/kg), mitomycin C (400 microgram/kg) or AG60 (5 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-3H-thymidine through tail vein. Seventy minutes after the thymidine injection, animals were sacrificed. The number of the labeled epithelial cells of the rectal crypts (mean number of labeled epithelial cells per 3.5 mm length of mucosa) were observed and calculated. On histological study, in the rectum of mitomycin C treated groups, narrowed intestinal gland, a number of the nectotic changed epithelial nuclei and loosely arranged lamina propria were observed. But in the AG60 treated group, morphological changes of the rectum were not observed. On autoradiographic study, number of the labeled cells of normal control, tumor control, 5-fluorouracil (30 mg/kg) treated, mitomycin C (400 microgram/kg) treated and AG60 (5 mg/kg) treated groups were 246.3+/-42.30, 253.8+/-20.54, 172.7+/-19.02, 108.7+/-17.67 and 53.8+/-11.70, respectively. In the AG60 and mitomycin C treated group, poorly-labeled cells containing only a few silver grains of 3H-thymidine were observed more frequently than in those of the normal control group. From the above results, AG60 (5 mg/kg) and mitomycin C (400 microgram/kg) are more suppressed the DNA synthesis of the cells of the rectal crypts as compare with 5- fluorouracil (30 mg/kg). And AG60 does not result any histological defect on the rectal mucosa. These results suggest that AG60 is expected as one of most effective anticancer drugs.

Ultrastructure of Rabbit Pyloric Mucosa Following Ligation of Common Bile Duct / 체질인류학회지

The purpose of this study was to examine the ultrastructural characteristic of the normal pylorus mucosa, and their structural changes induced by the ligation of common bile duct of the male rabbits weighing about 1.5 kg each. Experiment animals were divided into normal, sham operation, and experimental groups. Common bile duct ligation was performed under ether anesthesia and anjmals were sacrificed on the 1st, 3rd, 5th, 7th and 14th day after operation. The mucosal specimen of the pylorus, were fixed and embedded with common method. The sections were cut on a LKB-V ultratome, and observed under a JEM 100CX II electron microscope. The results were as follow : 1. In the early stages (1st, 3rd, 5th day groups) following the ligation, surface mucous cells have the various electron densities and shape of the mucous granules. In the late stages (7th, 14th day groups) following the ligation, many surface mucose cells containing numerous electron dense mucous granules are seen. 2. In the early stage of the ligation of bile duct, secretory function of EC cells was depressed, but in the later stage, the cells showed recovered secretory activity. 3. Secretory function of D cells was depressed on the early groups after the ligation of common bile duct, but they showed recovered secretory activity from the late groups after the ligation of the common bile duct. 4. Secretory function of G cells was activated on the early groups after the ligation of common bile duct, but they showed depressed secretory activity from the late groups after the ligation of the common bile duct. Considering the above findings, common bile duct ligation probably causes the dysfunction of the pyloric surface mucous cells that results in delayed mucous formation and secretion, and recovered mucous secretory function on the late stages. EC cells and G cells, depressed the secretory activities on the early stages and recovered on the late stages of the ligation of common bile duct. But D cells in the pyloric mucosa was activated on the early groups after the ligation of common bile duct ligation, but they was depressed secretory activities on the late groups.

Ultrastructural Alterations in the Gastric Chief Cells of Mouse, induced by 5-Fluorouracil or Mitomycin C / 대한해부학회지

This experiment was performed to evaluate the morphological responses of the gastric chief cells of the mouse, inoculated with Ehrlich carcinoma cells in the inguinal area, following administration of 5-fluorouracil or mitomycin C. Healthy adult ICR mice weighing 25 gm each were divided into normal and experimental groups (experimental control group, 5-fluorouracil-treated group and mitomycin C-treated group). In the experimental group, 1x107 Ehrlich carcinoma cells were inoculated subcutaneously in the inguinal area. From next day after inoculations, 0.2mL of saline (experimental control group), 5-fluorouracil (30 mg/kg, 5-fluorouracil-treated group), or mitomycin C (400 microg/kg, mitomycin C-treated group) were injected subcutaneously to the animals every other day, respectively. The day following the 7th injection, animals were sacrificed. Pieces of the tissue were taken from the gastric mucosa, prefixed with 2.5% glutaraldehyde-1.5% paraformaldehyde solution, followed by post-fixation with 1% osmium tetroxide solution. The ultrathin sections were stained with uranyl acetate and lead citrate. The size of zymogen granule and the size of the mitochondrion in the gastric chief cells were observed and compared. In the 5-fluorouracil treated group, most chief cells did not show any difference in ultrastructure, except myelin figures were more frequently observed, in comparison with those of normal control group. But in the mitomycin Ctreated group, necrotic cells were more frequently observed than in normal control and 5-fluorouracil-treated group. The size of zymogen granule in the gastric chief cells of normal control, experimental control, 5-fluorouracil-treated and mitomycin C-treated groups were 0.98 (+/-0.108)microm, 1.05 (+/-0.092)microm, 0.94 (/-0.123)microm and 0.93 (+/-0.156)microm, respectively. And the size of mitochondrion in the gastric chief cells of normal control, experimental control, 5-fluorouracil-treated and mitomycin C-treated groups were 0.80 (+/-0.130)microm, 0.83 (+/-0.143)microm, 0.87 (+/-0.165)microm and 0.81 (+/-0.083)microm, respectively. From the above results, in the treatment of low therapeutic doses of anticancer drugs into the animals inoculated with Ehrlich carcinoma cells, 5-fluorouracil may not suppress function of the gastric chief cells, but mitomycin C may exert a vicious influence on the function of the gastric chief cells.

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

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 5-fluorouracil, mitomycin C or CP -2. Healthy adult ICR mice weighing 25 gm each were divided into normal and experimental groups. In the experimental groups, each mouse was inoculated with 1 x10(7) Ehrlich carcinoma cells subcutaneously in the inguinal area. From the next day of inoculation, 0.2 mL of saline (experimental control group), 5-fluorouracil (30 mg/kg), mitomycin C (400 microgram/ kg) or CP -2 (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/g of methyl -(3)H-thymidine (25 Ci/mmol) 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 mitomycin C treated groups, narrowed intestinal gland, a number of the nectotic epithelial nuclei and loosely arranged lamina propria were observed. However, in the CP-2 treated group, morphological changes of the duodenum were not observed. On autoradiographic study, number of the labeled cells of normal control, experimental control, CP-2 treated, 5-fluorouracil treated and mitomycin C treated groups were 625.5 +/-58.85, 691.3 +/-82.32, 428.3 +/-83.16, 527.5 +/-79.84 and 297.33 +/-45.72, respectively. In the CP-2 and mitomycin C 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. From the above results, CP-2 and mitomycin C are more suppressed the DNA synthesis of the cells of the duodenal crypts as compare with 5-fluorouracil. But CP-2 does not result any histological defect on the duodenal mucosa. These results suggest that CP-2 is expected as one of most effective anticancer drugs.

Immunohistochemical Study of Nitric Oxide Synthase in Lacrimal Gland and Salivary Gland / 대한해부학회지

Endogenous nitric oxide (NO) has been known to regulate the salivary secretion and glandular blood flow. However, nitric oxide synthase (NOS) responsible for NO synthesis has not been well studied in lacrimal glands. The present study was aimed to investigate the distribution of nitric oxide synthase isoforms (endothelial, neuronal, and inducible NOS). Immunohistochemistry, using monoclonal mouse anti-endothelial NOS, anti-neuronal NOS, and anti-inducible NOS, was performed in exorbital lacrimal glands of the rat. Endothelial NOS (eNOS)-positive immunoreactivity was observed in vascular endothelium, intralobular duct and interlobular duct of the exorbital lacrimal gland of the rats, and also in the 3 major salivary glands of the rat. eNOSpositive immunoreactivity was most prominent in the intralobular and interlobular duct was well concentrated in cytoplasm of columnar epithelial duct cell. However, eNOS-positive immunoreactivity of the intercalated duct and serous acinus was absent. Neuronal NOS (nNOS)-positive immunoreactivity was seen in ganglion cells of exorbital lacrimal gland. iNOS or nNOS-positive immunoreactivy was not detected either in excretory ducts or in acinar cells. Inducible NOS-positive immunoreactivity was not seen. There results reveal the presence of eNOS and nNOS in the exorbital lacrimal gland, which may be related with regulation of the glandular secretion and blood flow.

Ultrastructure of Fronto-Accumbens Synapses after Prefrontal Decortication of the Rat / 대한해부학회지

Prefrontal cortex is called psycological cortex, since it deals with making up of individual personality, regulation of personal depth of feeling, working memory, planning, maintaining attention, etc. Whereas, nucleus accumbens (septi) is called the center of reward and motivation or the center of pleasure, since it deals with feeding, drinking, sex, exploration, appetitive learning, drug addiction, etc. Present study was aimed at the proving the prefronto-accumbens input ultrastructurally. Sprague Dawley rats anesthetized with sodium pentobarbital and were removed their prefrontal cortex with suction instrument. Two days following the operation, heads of rats were fixed by perfusion of with 1% glutaraldehyde-1% paraformaldehyde solution via left ventricle. Peristaltic pump was used during perfusion. Two hours later, brains were removed and refixed for 24 hours in the refrigerator, and small tissues of the nucleus accumbens were punched out with punching needle. Tissue blocks were fixed in 2% osmic acid for 2 hours and were embedded in araldite mixture. Ultrathin sections stained with uranyl acetate-lead citrate solution were observed with JEOL 100 CX II electron microscope. In the nucleus accumbens, some axodendritic terminals and axospinous terminals were found degenerated, and volume of activated glial cytoplasm was increased. The degenerated terminals were seen isolated from intact structures by activated glial processes and removed by glial cytoplasm. The result confirms that axon terminals coming from prefrontal cortex input to the spiny neurons of nucleus accumbens septi, on their dendrites and/or dendritic spines.

Effects of BCG or AG60 on Gastric Parietal Cells of the Mouse Implanted with Ehrlich Carcinoma Cells / 대한해부학회지

This experiment was performed to evaluate the morphological responses of the gastric parietal cells of mouse inoculated with Ehrlich carcinoma cells, following administration of Bacillus Calmette -Guerin (BCG) or acriflavine -guanosine composition (AG60, Taerim Pharm. Co. Seoul, Korea). In the experimental groups, each mouse was inoculated with 1 X 10(7) Ehrlich carcinoma cells subcutaneously in the inguinal area. From next day, 0.2 ml of saline, BCG (0.03 X 10(8) ~0.32 X 10(8) CFU) or AG60 (30 mg/kg) was injected subcutaneously to the animals every other day. Animals were sacrificed on the 14th day from 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 stained with uranyl acetate and lead citrate were observed with a JEM 100CX -II electron microscope. In the experimental control, the BCG and the AG60 treated groups, most parietal cells showed reduced lumenal spaces of the intracellular canaliculi, since microvilli of intracellular canaliculi were very irregularly shaped and crowed with each other. And in the BCG and the AG60 treated mice, myelin figures, lysosomes and multivesicular bodies in the parietal cells were observed more frequently than in those of the experimental control ones. In the BCG treated rats, membranes of the tubulovesicles of the parietal cells were disintegrated, but the similar changes were not observed in the AG60 treated mice,. Above results suggest that the BCG treated animals inoculated with Ehrlich carcinoma cells might suffer from reduced acid secretion of the parietal cell, since the disintegration of the tubulovesicular membranes in the parietal cells are occurred following injections. Whereas AG60 dose not affect remakably defect on the parietal cells.

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.

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 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.

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.