• Applied Microscopy
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• v.15 no.2
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• pp.111-129
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• 1985

The morphology and histology of the gastrointestinal mucosa of the mole, Talpa micrura coreana (Thomas), were studied using light and scanning electron microscopes. Tissue specimens were taken from body and pyloric portions of the stomach, and from the initial, proximal, middle, distal and terminal portions of the intestine. For light microscopy, tissue blocks were fixed in 10% buffered neutral formalin, embedded in paraffin wax, and sectioned at a thickness of $5{\mu}m$. These sections were stained with hematoxylin-eosin. For scanning electron microscopy, tissue blocks were fixed in 1% glutaraldehyde-1.5% paraformaldehyde, and postfixed in 1% osmium tetroxide, dehydrated in graded alcohol, transferred to isoamylacetate and dried by the critical point drier(Polaron E 3000). Subsequently, specimens were coated with gold and observed with a JSM-35C scanning electron microscope. The results were as follows: The mucous membrane of the body portion of the stomach had numerous irregular folds and the pyloric mucosa formed the strawberry-shaped folds, and general histological structures of each portion were similar to those of man. The intestine could not be differentiated macroscopically and microscopically into small and large intestines. There was no cecum, appendix, taenia coli, haustra coli or appendices epiploicae. In the initial portion (4 mm long), conical or tongue-shaped villi with the height of $143.3{\pm}10.7{\mu}m$ were present, and large mucous glands were seen in the submucosa. In the proximal, middle and distal portions, wavy folds composed of the epithelium and lamina propria were densely and transversely arranged, and their heights were $440.4{\pm}45.5{\mu}m,\;454.4{\pm}19.9{\mu}m\;and\;205.2{\pm}33.5{\mu}m$, respectively. The mucosa of the terminal portion (3 cm long) formed several longitudinal folds, and the intestinal glands were directly opened on the smooth surface of the folds. Aggregated lymphoid follicles were observed in the major portions of the intestine except the initial and terminal portions. There was no circular or semilunar fold throughout the intestine.

• Applied Microscopy
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• v.23 no.2
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• pp.53-77
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• 1993

In this study, an attempt was made to investigate the probable organelles participating in the secretion of biligrafin. The animals (ICR male mice, 25-30gm) were divided into normal control and 6 biligrafin injected groups to which 30% biligrafin (0.006ml/gm b.w.) were injected at 10, 20, 40, 80, 160 and 320 min prior to the sampling. The mice of each group were perfused through the heart with ice-cold 2.5% glutaraldehyde buffered with 0.1M Na-cacodylate (pH. 7.4) under the Na-pentobarbital (Nembtal 0.0015mg/gm b.w.) anesthesia and liver tissues were taken from each group. Some specimens were immersed 1 hr in the same solution used in the perfusion. After an overnight rinse in 0.1M Na-cacodylate buffer containing 10% DMSO and 7.6% sucrose, $75{\mu}m$ fronzen sections were made for cytochemical study. The sections were incubated in thiamin pyrophosphatase (TPPase) and inosine diphosphatase (ID Pase) media for 70 min at $37^{\circ}C$ respectively and acid phosphatase (AcPase) medium for 40 min at $37^{\circ}C$. They were postfixed in 1 % $OsO_4$ for 1 hr. The other specimens were immersed for 8 hrs in the fixative consisting of 2.5% glutaraldehyde and 3.0% paraformaldehyde buffered with Na-cacodylate (pH. 7.4). All of the osmificated specimens were processed for electron microscopy. In both normal and biligrafin injected groups, endoplasmic reticulum (ER), vacuoles, Golgi apparatus and lysosomes were seen in the vicinity of bile canaliculus. In the biligrafin injected groups, however, the Golgi apparatus appeared to be decreased and ER and vacuoles were dilated and increased. The rough endoplasmic reticulum (RER) having a few attached ribosomes appeared to be the round saccule, especially at 20 min after biligrafin injection. Smooth endoplasmic reticulum (SER) seemed to be formed by the detachment of ribosomes at the cisternal end of RER. The cistern of SER showed saccules which probably budded off to form the vacuole. The vacuoles were devoid of visible centents. This finding seemed to be in agreement with the biochemical property of the bile constituents. The fusion between the vacuoles and bile canaliculus were frequently seen in the groups injected with biligrafin. The lysosome did not show any changes in the biligrafin injected groups. Accumulation of some material and lipid droplets were seen at the 40 and 80 min after biligrafin injection, especially at the latter. At 160 and 320 min after biligrafin injections, however, they were decreased successively while the RER stack, free ribosomes and polysomes were increased. Although the reactive products of TPPase and IDPase were observed in the ER saccules and vesicles of the normal control and biligrafin injected groups, the fusion between the bile canaliculus and saccules or vesicles could easily be seen in the latter. The AcPase activity, however, was observed in the cistern at the maturing face of Golgi apparatus and lysosomes in both normal and biligrafin groups. The results suggest that the biligrafin is excreted via the vesicles, vacuoles or sacoules probably derived from the SER without the participation of Golgi apparatus and lysosomes, and the excess amount of material is stored as inclusions during the repairing of the organelles being overactive.

• Applied Microscopy
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• v.29 no.1
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• pp.57-66
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• 1999

The nervous tissue in the cerebral ganglion of Korean planaria was observed using electron microscope. The obtained results are as follows: A cerebral ganglion is composed of the nerve cells, neurosecretory cells, neuroglial cells and neuropils. The nerve cells are round or ovoidal-shaped cells (diameter, $5{\mu}m$), which has a large ellipsoidal nucleus containing the evenly developed heterochromatin. Their cytoplasms were found to be relatively simple, because of their undeveloped cell organelles. The neurosecretory cells are long and ellipsoid or spindle-shaped cells, where there were found a large ellipsoidal nucleus and cytoplasm filled with secretory granules (diameter, 60 nm). The neuroglial cells were seldom observed. They are spindle-shaped cells (size, $6\times0.8{\mu}m$), which were observed mainly among the nerve fibers. The neuropils are formed by the nerve fibers and nerve endings which are filled with mitochondria, neurotubules and secretory granules of four kinds (high electron dense granules of sizes 75 nm, 50 nm and 37 nm, and electron lucent granule of size 30 nm etc.). These granular vesicles are divided into single vesicle type and compound vesicle type in the nerve terminals, and neuronal synapses were observed to be the axo-dendritic and dendro-dendritic synapse type.

• Applied Microscopy
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• v.28 no.4
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• pp.425-434
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• 1998

This study was designed to investigate the microglial reactions to the neurodegenerative changes in the cat retina. All experiments were performed using adult cats of both sex, weighing $2,500g\sim3,500g$. 5,7-DHT $(100{\mu}g)$ dissolved in 0.1% ascorbic acid was injected into the vitreous body. All injections were performed in one-side eye; the other side served as the control, which was injected only with 0.1% ascorbic acid. Cats were sacrificed at 1, 3, 7, 14 and 21 days after intravitreal injection of 5,7-DHT For light microscopy, retinae were fixed with 4% paraformaldehyde and processed using NDPase histochemistry. Same retinae were fixed with 1% para(formaldehyde-2.5% glutaraldehyde and processed for electron microscopy. NDPase-positive microglial cells were mainly distributed in the inner plexiform layer of the retina, and characterized by a small somata with a few slender processes, which were also extended in the ganglion cell layer (GCL) and inner nuclear layer (INL). The intensity of the microglia stained for NDPase was abruptly increased at 7 day as compared with that of the control, and thereafter continuously sustained until 21 day, the last experimental group in this study. Under the electron microscopical observation, microglial cells in the control group exhibited elongate nucleus with perinuclear chromatin condensation, and the perikaryon was scanty. However, a few hypertrophic glial cells were frequently found at 3 days after the drug injection. By 7 day, most microglial cells directed toward the degenerated neurons in the GCL, and the number of microglial cells was slightly increased as compared with the former group. At the 14 day, most microglial cells wrapped the degenerated cells in the GCL, and a few cells showed phagocytotic features. By 21 day, most microglial cells were engaged in phagocytotic activity, and their cytoplasm was filled with the phagorytosed material. Based on the results, 5,7-DHT may act as a specific neurotoxin to the cat retina, and microglial reactions to the neuronal death are already induced in early experimental stage. These results indicate that the microglial cells in the cat retina show characteristic features as a protective effect of neural tissue.

• Restorative Dentistry and Endodontics
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• v.24 no.2
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• pp.372-380
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• 1999

The purpose of this study was to investigate the distribution of calcitonin gene-related peptide(CGRP) containing nerve fivers after pulp exposure in rats. The Spague-Dawley rats weighing about 250 - 300g were used. The animals were devided into normal control group and experimental groups. Experimental animals were sacrified on 2, 4, 7, 10 days after pulp exposure. The maxillary teeth and alveolar bone were removed and immersed in the 4% paraformaldehyde plus 0.1M phosphate buffer (pH 7.4). Serial frozen $50{\mu}m$ thick sections were cut with a cryostat. In the immunohistochemical staining procedure, the rabbit CGRP antibody was used as a primary antibody. The sections were incubated for 48 hours at $4^{\circ}C$, and placed into biotinylated anti-rabbit IgG as a secondary antibody and incubated in ABC (avidin-biotin complex), The sections were visualized by 0.05% 3.3 diaminobenzidine tetrahydrochloride. The results of this study were as follows: 1. In control group, CGRP containing nerve fibers ran parallel to the long axis of root and reached the coronal pulp. They were distributed on Raschkow plexus under the odontoblastic layer. 2. In 2 day group after pulp exposure, tissue necrosis and acute inflammation occurred and CGRP containing nerve fibers increased. In 4 day group, the necrotic tissue extended to the pulp and CGRP containing nerve fibers were distributed around the inflammation zone. 3. In 7 day group after pulp exposure, pulp necrosis occurred, and in 10 day group, the abscess under the necrotic pulp extended to the root apex area and CGRP containing nerve fibers were not observed in root canals. 4.The sprouting of CGRP nerve fibers was most remarkable at the pulp chamber under injury in 4 day group, and it was found at inflammation zone under the necrotic tissue in 7 day group and the remaining root pulp tissue in 10 day group. As mentioned above, CGRP nerve fibers had a tendency to increase around the inflammatory zone, especially around the acute inflammation tissue, when compared with control group. It is suggested that CGRP nerve fibers maybe related to the control of inflammatory response of pulp tissue.

• Restorative Dentistry and Endodontics
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• v.25 no.2
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• pp.225-234
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• 2000

The purpose of this study was to investigate the distribution and fluorescene intensity of vasoactive intestinal polypeptide(VIP) immunoreactive cells in rat trigeminal ganglion after inferior alveolar nerve axotomy. The animals were divided into normal and two experimental groups. The experimental animals were sacrificed at 14th and 28th day after inferior alveolar nerve axotomy. The trigeminal ganglion was removed and immersed in the 4% paraformaldehyde-0.2% picric acid in 0.1M phosphate buffer. Serial frozon sections about $16{\mu}m$ in thickness were cut with a cryostat. The immunofluorescence staining was performed. The rabbit anti-VIP(1 : 8,000) was used as primary antibody and fluorescene isothiocynate(FITC)-conjugated anti-rabbit IgG(1 : 80) as secondary antibody. The slides were observed under confocal laser scanning microscope. Three-dimensional images were constructed from 9 serial images(each $1{\mu}m$ in thickness) made by automatic optical sectioning. Unprocessed optical sections were obtained and stored on a optical disk. Color picture were printed by a video copy processor. The results were as follows; 1. The appearance of VIP immunoreactive cells in the mandibular part of trigeminal ganglion was 8.79${\pm}$1.99% in normal group and 39.16${\pm}$5.62% in 14 days, 16.25${\pm}$2.39% in 28 days after inferior alveolar nerve axotomy groups. 2. The relative fluorescence intensity of VIP immunoreactive cell bodies in the mandibular part of trigeminal ganglion was 134.40${\pm}$10.39 in normal group and 192.88${\pm}$14.06 in 14 days, 143.10${\pm}$5.02 in 28 days after nerve axotomy groups. Therefore, the relative fluorescence intensity of 14 days after nerve axotomy group was 43.3% higher than intensity of normal group. 3. In optical single section analysis of VIP immunoreactive cell bodies, white cell bodies(moderate fluorescence intensity) were the most abundant in normal and 28 days after nerve axotomy groups. Whereas, in 14 days after nerve axotomy group, red cell bodies(high fluorescence intensity) were the most abundant. 4. In optical serial section analysis of VIP immunoreactive cell bodies, red cell bodies(high fluorescence intensity) were observed in a part of the 9 sections of normal and 24 days after nerve axotomy groups. Whereas, red cell bodies were observed in all of the 9 sections of 14 days after nerve axotomy group. 5. The results indicates that number and fluorescence intensity of VIP immunoreactive cells were increased in the mandibular part of trigeminal ganglion following inferior alveolar nerve axotomy.

• The Journal of the Korea Contents Association
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• v.19 no.7
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• pp.186-191
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• 2019

Introduction: In aged people, stroke incidence is increased. But standardized experimental animal protocol study for the research of stroke therapy is rare. There is little report on the success rate of cerebral artery occlusion model using standardized Nylon thread length of precise thread end-size controlled. Method: In this study, the operator intended the occlusion of middle cerebral artery (MCA) using $0.18{\pm}0.02mm$ end 5-0 Nylon thread. Middle cerebral artery occlusion was induced for 60min under isoflurane anesthesia. After 60min, the operator removed the Nylon thread and reperfusion was induced for 23hrs. The mice was killed 23hrs after reperfusion and infarction area of brain was confirmed by 1.5% TTC (2,3,5-tryphenyl tetrazolium chloride) staining. Results: According to end size and insert length of Nylon thread, Middle cerebral artery occlusion (n=50), internal carotid artery occlusion (n= 14), distal middle cerebral artery occlusion (n= 36), anterior cerebral artery (n= 1) were induced. And no infarction (n= 50) was observed. Conclusion: According to weight of mice, the operator induced reversible cerebral artery occlusion model by different insert length (30.0~36.9g : 9.0mm, 37.0~40.0g : 9.5mm) of Nylon thread. Success of cerebral artery occlusion model was confirmed by checking infarction area using TTC staining. The success rate (66.9%, 101/151) of reversible cerebral artery occlusion model in the mouse and the operational conditions are shown.

• 대한예방의학회:학술대회논문집
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• 1994.02b
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• pp.169-179
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• 1994

B-mode ultrasound is being used to assess carotid atherosclerosis in epidemiological studies and clinical trials. Recently the interpretation of measurements made from ultrasound images has been questioned. This study examines the anatomical correlates of B-mode ultrasound of carotid arteries in vitro and in situ in cadavers. Twenty-seven segments of human carotid artery were collected at autopsy. pressure perfusion fixed in buffered 2.5% gluteraldehyde and 4% paraformaldehyde and imaged using an ATL UM-8 (10 MHz single crystal mechanical probe). Each artery was then frozen, sectioned and stained with van Gieson or elastin van Gieson. The thickness of the intima. media and adventitia were measured 'to an accuracy of 0.01 mm from histological sections using a calibrated eye graticule on a light microscope. Shrinkage artifact induced by histological preparation was determined to be 7.8%. Digitised ultra sound images of the artery wall were analysed off-line. The distance from the leading edge of the first interface ($LE_{1}$) to the leading edge of the second interface ($LE_2$) was measured using a dedicated programme. $LE_{1}$-$LE_{2}$ measurements were correlated against histological measurements corrected for shrinkage. Mean values for the far wall were: ultra sound $LE_{1}$-$LE_{2}$ (0.97 mm, S.D. 0.26), total wall thickness (1.05 mm, S.D. 0.37), adventitia (0.35 mm, S.D. 0.16), media (0.61 mm, S.D. 0.18). intima (0.09 mm, S.D. 0.13). Ultrasound measurements corresponded best with total wall thickness, rather than elastin or the intima-media complex. Excision of part of the intima plus media or removal of the adventitia resulted in a corresponding decrease in the $LE_{1}$-$LE_{2}$ distance of the B-mode image. Furthermore. increased wall thickness due to intimal atherosclerotic thickening correlated well with $LE_{1}$-$LE_{2}$ distance of the B-mode images. B-mode images obtained from the carotid arteries in situ in four cadavers also corresponded best with total wall thickness measured from histological sections and not with the thickness of the intima plus media. In conclusion, the $LE_{1}$-$LE_{2}$ distance measured on B-mode images of the carotid artery best represents total wall thickness of intima plus media plus adventitia and not intima plus media alone.

• The Korean Journal of Zoology
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• v.21 no.1
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• pp.29-39
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• 1978

The authors observed the ultrastructure of the mucous glandular epithelial cells in the amphibian skin by mean of electron microscope. The specimens from the experimental animals were fixed in 2.5% glutaraldehyde-oaraformaldehyde fixative in phosphate buffer at pH 7.2 prior to fixation in 1% osmium tetroxide, dehydrated in graded ethanol and acetone, embedded in Epon 812 mixture, and sectioned with Sorvall MT-2 ultramicrotome. The ultrasections were contrasted with uranyl acetate and lead citrate and observed with a JEOL-100B electron microscope. The results were as follows: 1. The cutaneous mucous glands in amphibia consisted of the glandular epithelial and the myoepithelial cells. 2. Several different cells in ultrastructure were observed in the mucous glandular epithelium of the adult amphibian skin. a. The dark and the light cells were observed in Hynobius leechi. b. The mitochondria-rich and the round secretory granule-containing cells were observed in Bombina orientalis. c. The round secretory granule-containing and the foam-like granule mass-containing cells were observed in Kaloula tornieri. d. The cutaneous mucous gland of Rana nigromaculata were divided into two types: A and B-type glands. In the A-type mucous gland, the mitochondria-rich and the round secretory granule-containing cells and in the B-type mucous gland, the mitochondria-rich, the secretory granule-containing and the ER-rich cells were observed. 3. Based upon the above findings, the authors infer that the mucous granular epithelium of the amphibian skin consists of the mitochondria-rich undifferentiated, the secetory granule-containing and mature, and the ER-rich evacuated cells.

• Restorative Dentistry and Endodontics
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• v.27 no.4
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• pp.355-362
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• 2002

One fifth dilution of formocresol is usually used for pulpotomy of the primary teeth and emergency pulpotomy of the permanent teeth. However the use of formaldehyde has been subjected to criticism because it may be absorbed into the blood stream and become distributed systemically, it nay also alter the pulp tissue rendering it immunologically active, and have carcinogenic potential. Recently Depulpin$^{\circledR}$(VoCo., Germany) gains popularity as a devitalizing agent during root canal therapy in spite of high concentration of 49 % paraformaldehyde because it facilitate devitalization of pulp and make root canal therapy easier But there have been not enough publications about the reaction of pulp and periapical tissue caused by Depulpin. This study was performed to evaluate the histological changes in pulp and periapical tissue of rats after pulpotomy using formocresol and Depulpin and to elucidate the toxic effects of these agents. Thirty six Sprague-Dawley rats were anesthetized by intraperitoneal injection of ketamine Maxillary first molar teeth were used for pulpotomy with formocresol and Depulpin. Rats were sacrificed after 2 days, 4 days, 1 week, 2 weeks, 3 weeks and 4 weeks respectively. Specimens were histologically observed by light microscope changes in pulp and periapical tissue. The obtained results were as follows. 1. Formocresol group A zone of fixed tissue. in which odontoblasts could clearly be defined, was present directly underneath the pulpotomy dressing in almost all teeth of this group. This was followed by an area of necrotic tissue which resembled dried out fibrous tissue with no cellular detail except some pyknotic nuclei. In the specimens of after 2 days, 4 days, 1 week, 2 weeks in which vital tissue was present, it was separated from the fibrous area by a zone of inflammation. In the specimens of after 3 weeks and after 4 weeks, inflammatory infiltrate was in the periodontal ligament adjacent to the apical foramina of the teeth. 2. Depulpin$^{\circledR}$ group The area of necrotic tissue which had no cells and fibers, was present adjacent to the dressing. This was followed by dried out fibrous tissue with no cellular details except some pyknotic nuclei, A short stump of vital pulp with odontoblasts was present at the end of the canal after 2 days. Inflammatory infiltrate was in the periodontal ligament after 4 days and after 1week. Severe root resolution and necrosis of periapical tissue opposite the root resorption site were defined after 2 weeks and after 3 weeks. Periapical lesion which consist of necrotic tissue surrounded by a fibrous connective wall, was found after 4 weeks. The results indicated that Depulpin can cause more adverse reaction to the dental pulp and periapical tissue than formocresol, and further studies are needed for its clinical use with safety.