• Journal of Physiology & Pathology in Korean Medicine
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• v.23 no.4
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• pp.805-817
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• 2009

This study was to investigate central localization of neurons projecting to the urinary bladder and urinary bladder-related acupoints(Wijung, $BL_{40}$) and neurons of immunoreactive to hormones and hormone receptors regulating urinary bladder function by using peudorabies virus(PRV). In this experiment, Bartha's strain of pseudorabies virus was used in rats to trace central localization of urinary bladder-related neurons and urinary bladder-related acupoints($BL_{40}$) which can regulate urinary system. PRV was injected into the urinary bladder and acupoints($BL_{40}$) related urinary system. After six days survival of rats, mainly common labeled neurons projecting to the urinary bladder and urinary bladder-related acupoints were identified in spinal cord, medulla, pons and diencephalon by PRV immunohistochemical staining method. First-order PRV labeled neurons projecting to urinary bladder and urinary bladder-related acupoints were found in the cervical, thoracic, lumbar and sacral spinal cord. Commonly labeled preganglionic neurons were labeled in the lumbosacral spinal cord and thoracic spinal cord. They were found in the lateral horn area(sacral parasympathetic nucleus and intermediolateral nucleus), lamina V-X, intermediomedial nucleus and dorsal column area. The area of sensory neurons projecting to urinary bladder and Wijung($BL_{40}$) was L5-S2 spinal ganglia and T12-L1 spinal ganglia, respectively. In the brainstem, the neurons were labeled most evidently and consistently in the nucleus of tractus solitarius, area postrema, dorsal motor nucleus of vagus nerve, reticular nucleus, raphe nuclei(obscurus, magnus and pallidus), C3 adrenalin cells, parapyramidal area(lateral paragigantocellular nucleus), locus coeruleus, subcoeruleus nucleus, A5 cell group, Barrington's nucleus and periaqueductal gray matter. In the diencephalon, PRV labeled neurons were marked mostly in the paraventricular nucleus and a few ones were in the lateral hypothalamic nucleus, posterior hypothalamic nucleus, ventromedial hypothalamic nucleus, arcuate nucleus, median eminence, perifornical nucleus, periventricular nucleus and suprachiasmatic nucleus. In cerebral cortex, PRV labeled neurons were marked mostly in the frontal cortex, 1,2 area, hind limb area, agranular insular cortex. Immunoreactive neurons to Corticotropin releasiing factor(CRF), Corticotropin releasiing factor-receptor(CRF-R), c-fos and serotonin were a part of labeled areas among the virus-labeled neurons of urinary bladder and Wijung($BL_{40}$). The commonly labeled areas were nucleus tractus solitarius, area postrema, reticular nucleus, raphe nuclei(obscurus, magnus and pallidus), locus coeruleus, A5 cell group, Barrington,s nucleus, arcuate nucleus, paraventricular nucleus, frontal cortex 1, 2 area, hind limb, and perirhinal(agranular insular) cortex. These results suggest that overlapped CNS locations are related with autonomic nuclei which regulate the functions of urinary bladder-relate organs and it was revealed by tracing PRV labeled neurons projecting urinary bladder and urinary bladder-related acupoints. These commonly labeled areas often overlap with the neurons connected with hormones and hormone receptors related to urination.

• The Journal of Korean Medicine
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• v.18 no.1
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• pp.58-71
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• 1997

In order to the location and local arrangement of nerve cell bodies and nerve fibers projecting to the meridian points related to facial nerve paralysis in the rat using the neural tracers, CTB and WGA-HRP, labeled neurons the were investigated by immunohistochemical and HRP histochemical methods following injection of 2.5% WGA-HRP and 1% CTB into Hyopko$(S_6)$. Chichang$(S_4)$, Sugu$(GV_{26})$, Sajukkong$(TE_{23})$ and Yangbaek$(G_{14})$. Following injection of Hyopko$(S_6)$, Chichang$(S_4)$, labeled motor neurons were founded in facial nucleus, trigeminal motor nucleus, reticular nucleus and hypoglossal nucleus. labeled sensory neurons were founded in trigeminal ganglia and $C_{1-2}$ spinal ganglia. sympathetic motor neurons were found in superior cervical ganglia. Sensory fibers labeled in brainstem were found in mesencephalic trigeminal tract, sensory root of trigeminal nerve, oral, interpolar and caudal part of trigeminal nucleus, area postrema, nucleus tractus solitarius, lateral reticular nucleus and $C_{1-2}$ spinal ganglia. Following injection of Sugu$(GV_{26})$, labeled motor neurons were founded in facial nucleus. Labeled sensory neurons were founded in trigeminal ganglia and $C_{1-2}$ spinal ganglia. Sympathetic motor neurons were found in superior cervical ganglia. Sensory fibers labeled in brainstem were found in spinal trigeminal tract, trigeminal motor nucleus, mesencephalic trigeminal tract, oral. interpolar and caudal parts of trigeminal nucleus, area postrema, nucleus tractus solitarius, lateral reticular nucleus, dorsal part of reticular part and $C_{1-2}$ spinal ganglia. Following injection of Sajukkong$(TE_{23})$ and Yangbaek$(G_{14})$, labeled motor neurons were founded in facial nucleus, trigeminal motor nucleus. Labeled sensory neurons were founded in trigeminal ganglia and $C_{1-2}$ spinal ganglia. sympathetic motor neurons were found in superior cervical ganglia. Sensory fibers labeled in brainstem were found in oral, interpolar and caudal parts of trigeminal nucleus, area postrema, nucleus tractus solitarius, inferior olovary nucleus, medullary reticular field and lamina I-IV of $C_{1-2}$ spinal cord. Location of nerve cell body and nerve fibers projecting to the meridian points related to the facial nerve paralysis in the rats were found in facial nucleus and trigeminal motor nucleus. Sensory neurone were found in trigeminal ganglia and $C_{1-2}$ spinal ganglia. Sympathetic motor neurons were found in superior cervical ganglia. Sensory fibers labeled in brainstem were found in mesencephalic trigeminal tract, oral, interpolar and caudal parts of trigeminal nucleus, area postrema, nucleus tractus solitarius. lateral reticular nucleus, medullary reticular field.

• Development and Reproduction
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• v.4 no.1
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• pp.19-28
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• 2000

Diploid and triploid hybrid between female mud loach Misgurnus mizolepis and male cyprinid loach, M. andguillicaudatus were produced, using artificial propagation. The influence of hybridization and triploidization on characteristics of the some tissues of retina optic tectum and trunk kidney in loach was determined histologically. Diploid hybrid have almost size similarities in nucleus and the cell size of some tissues to both parents. In the triploid hybrid the nucleus and／or the cell size in erythrocyte, horizontal cell in inner nuclear layer of retina ganglion cell in ganglion cell layer of retina, neuronal cell in granular layer of optic tectum, and secondary proximal tubule cell of trunk kidney, are much larger than those of the corresponding cells in the diploid hybrid individuals. However, triploid hybrid showed fewer number of cells in outer nuclear layer in retina and nuclei in secondary proximal tubule of trunk kidney than in diploid hybrid. Results of this study suggest that some characteristics in the triploid hybrid exhibiting larger cell and nucleus size with fewer number of cell than diploid hybrid can be useful criteria for the distinction between diploid hybrid and triploid hybrid, and the ploidy level in loach.

• Applied Microscopy
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• v.25 no.1
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• pp.1-14
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• 1995

The purpose of this study was to investigate the main sensory trigeminal nucleus in the aging rat brain by means of electron microscope. Male Sprague-Dawley rats, two (control group) and thirty six (aging group) months of age, were used. These animals were sacrificed by perfusion fixation with 2.5% glutaraldehyde-2.0% paraformaldehyde (0.1M phosphate buffer, pH 7.4) under sodium pentobarbital. The objective area was punched out with a sharp-edged metal cylinder of 0.8 mm in diameter. These blocks of tissue were then washed in 0.1M phosphate buffer, postfixed in 2% osmium tetroxide, dehydrated in a graded series of ethyl alcohol, and embedded in Epon 812. Thin sections were cut with Super Nova ultramicrotome, pick up on grids and double stained with lead citrate and uranyl acetate, and observed in JEOL 100B electron microscope. The results were as follows: 1. In the control group, the neuronal cell body of the main sensory trigeminal nucleus was filled with nucleus, Golgi complex, Nissl substance, mitochondria, microfilaments and microtubules. However, few Nissl substances are seen in neuronal cell body. Axoaxonic synapse, axodendritic synapse, axosomatic synapse, axospinous synapse, myelinated and unmyelinated nerve fibers were well organized around cell bodies. Neurons with abnormal changes were not seen. 2. In the aging group, the neuronal cell body of the main sensory trigeminal nucleus contained large number of lipofuscin granules, dense body and swollen mitochondria. Terminal boutons contained glycogen, crystal-like vesicle and membranous indicating first signs of degeneration. The dendrites were found to be in synaptic contact with altered axon terminals. Frequently axons filled with dark axoplasn and splitted myelin sheath were noticed.

• Korean Journal of Microbiology
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• v.14 no.3
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• pp.135-145
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• 1976

Confluent AGMK cells were infected by large plaque SV40 virus. Levels of DNA polymeras $({\alpha}\;and\;{\beta})$ were measured in the cytoplasm and the cell nucleus. The activities of DNA $polymerase-{\alpha}$ which found in both the cell nucleus and the cytoplasm were increased approximately eight folds at 48 hours after infection of SV40 virus. Only insignificant but constant amounts of DNA $polymerase-{\beta}$ were found either in the nucleus of the SV40 infected cell or of the uninfected cell. The characteristics of the SV40 virus induced DNA polymerases were compared with that of the uninfected cellular DNA polymerase in regard of the effects of pH, salt concentration, NEM concentration and temperature on those enzyme activities. No differential effect was found between both enzymes. Endouclease activities wre examined in the purified DNA $polymerase-{\alpha}\;and\;{\beta}$. The low level of endonuclease activity which might cut SV40 DNA 1 at one site was observed in the DNA $polymerase-{\alpha}$ whereas high but nonspecific endonuclease activities were found in the DNA $polymerase-{\beta}$.

• Applied Microscopy
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• v.21 no.2
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• pp.39-56
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• 1991

The development of notochordal cells of nucleus pulposus was studied with electron microscope in human fetuses ranging from 30 mm to 260 mm crown-rump length. At 30 mm fetus, primitive notochordal cells were large with central nucleus, few organelles, and their cytoplasm usually contained dense glycogen and fine filaments. Notochordal cells at all ages contained bundles of fine filaments of indeterminate nature. One unusual feature of fetal notochordal cells was the consistent presense of rough endoplasmic reticulum surrounding poorly developed mitochondria. At 50 mm fetus, notochordal cells formed dense masses with interdigitating cell membranes connected by a variety of cell to cell junctions. With increasing age, the cell connections became slender threaded cytoplamic extending from cell and enclosed large extracellular space. Chondrocyte-like cells appeared to be separated by large volumes of extracellular matrix. Viable notochordal and condrocyte-like cells existed in specimen from all age. The extracellular spaces were filled with fibrillar and granular material by 90 mm fetus. Necrotic cells were distinguished by loss of their membrane integrity, vacuolization of their organelles, and the presence of dense osmiophilic masses. In adult tissue, notochordal cells became rounded or irregular in shape and developed a pericellular matrix consisting of collagen fibrile, and dense particle. The structure of notochordal cells and their persistance in the nucleus pulposus after fetal life suggested that they may have a significant role in the formation and maintenance of the nucleus pulposus. The presence of Golgi complex and well-developed endoplasmic reticulum in chondrocyte-like cells suggested that they are capable of producing and maintaining the extracellular matrix.

• Journal of Embryo Transfer
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• v.9 no.2
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• pp.153-160
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• 1994

This study evaluated the influence of cell stage of donor nucleus on nuclear injection, electrofusion and in vitro development in the rabbit to improve the efficiency of nuclear transplantation in the rabbit. The embryos of 8-, 16- and 32-cell stage were collected from the mated does by flushing viducts with Dulbecco's phosphate buffered saline(D-PBS) containing 10% fetal calf serum(FGS) at 44, 54 and 60 hours after hCG injection. The blastorneres separated from these embryos were used as donor nucleus. The ovulated oocytes collected at 14 hours after hCG injection were used as recipient cytoplasm following removing the nucleus and the first polar body. The separated blastomeres were injected into the enucleated oocytes by micromanipulation and were electrofused in 0.28 M mannitol solution at 1.5 kV /cm, 60 $\mu$sec for three times. The fused oocytes were cocultured with a monolayer of rabbit oviductal epithelial cells in M-199 solution containing 10% FGS for 72~120 hours at 39$^{\circ}C$ in a 5% $CO_2$ incubator. The cultured nuclear transplant embryos were stained with Hoechst 33342 solution and the number of cells were counted by fluorescence microscopy. The successful injection rate of 8-, 16- and 32-cell-stageblastomeres into enucleated oocytes was 86.7, 91.0 and 93.9%, respectively. The electrofusion rate of 8-, 16- and 32-cell-stage blastomeres with enucleated oocytes was 93.3,89.3 and 79.0%, respectively. Development of blastomeres to blastocyst was similar with 8-,16- and 32-cell-stage donor nuclei(26.2, 25.8 and 26.6%, respectively, P<0.05). The mean number of cell cycle per day during in vitro culture in nuclear transplant embryos which received 8-, 16- and 32-cell- stage nuclei was 1.87, 1.81 and 1.43, respectively.

• Applied Microscopy
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• v.44 no.1
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• pp.15-20
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• 2014

Cytokeratin 19 (CK19) expressed in epidermis of skin, bulge region of hair follicle, outermost layer of outer root sheath and proximal and distal to bulge. Vimentin is a fibrous protein that localized in cytoplasm of fibroblast and forms cytoskeleton to maintain shape of cell and nucleus. In this study, CK19 and vimentin in skin were confirmed with light, fluorescence and transmission electron microscope. As a result, CK19 was localized epidermis, hair follicles, outer root sheath and nucleus of Merkel's cell. However, vimentin was localized some epidermis, dermis, hypodermis and nucleus of Merkel's cell. The role of CK19 is self-renewal and homeostasis in skin. Also, hair follicle regeneration and hair growth is known to be related. It is supposed that required of structural proteins that make up cytoskeleton is increased. Thereby, expression of CK19 is increased. It is considered that vimentin localized in order to stabilize structure of cell and cytoskeleton of fibroblasts. Also, CK19 and vimentin present in nuclei of Merkel's cell, and to act as a fibrous protein that make up end of a nerve fiber present in Merkel's cell and paracrine function of Merkel's cell.

• Journal of Biomedical Engineering Research
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• v.10 no.2
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• pp.106-108
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• 1989

A new method of digital image analysis technique for discrimination of cancer cell was presented in this paper. The object image was the Thyroid eland cells image that was diagnosed as normal and abnormal (two types of abnormal: follicular neoplastic cell, and papillary neoplastic cell), respectively. By using the proposed region segmentation algorithm, the cells were segmented into nucleus. The 16 feature parameters were used to calculate the features of each nucleus. A9 a consequence of using dominant feature parameters method proposed in this paper, discrimination rate of 91.11% was obtained for Thyroid Gland cells.

• Korean Journal of Veterinary Research
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• v.37 no.3
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• pp.479-487
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• 1997

The present study was carried out to localize the central nuclei innervating the rat colon using pseudorabies virus-Bartha strain which has been known as a very useful neurotracer. The results were as follows. The central nuclei innervating the proximal colon were premotor area, subfornical organ, preoptic area in telencephalon, and paraventricular nucleus, bed nucleus of stria terminalis, retrochiasmatic area in the diencephalon, and periaqueductal gray, Edinger-Westphal nucleus, tegmental nucleus in the mesencephalon, and parabrachial nucleus, locus ceruleus, A5 area, $K{\ddot{o}}lliker$-Fuse nucleus, magnocellular reticular nucleus in the metencephalon, and nucleus tractus solitarius, A1 noradrenergic cell group, dorsal motor nucleus of vagus nerve, nucleus ambiguus, area postrema in the myelencephalon. In the spinal cord, the thoracic division had some nuclei innervating the proximal colon. The nuclei innervating the distal colon were paraventricular nucleus of the diencephalon, Edinger-Westphal nucleus of midbrain, and parabrachial nucleus, locus ceruleus, A5 area, $K{\ddot{o}}lliker$-Fuse nucleus, magnocellular reticular nucleus of the metencephalon, and nucleus tractus solitarius, dorsal motor nucleus of vagus nerve, nucleus ambiguus, area postrema in the myelencephalon. In the spinal cord, thoracic, lumbar and sacral division innervated the distal colon.