• Clinical and Experimental Reproductive Medicine
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• v.38 no.4
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• pp.216-221
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• 2011

Objective: To differentiate the human embryonic stem cells (hESCs) into the retinal pigment epithelium (RPE) in the defined culture condition and determine its therapeutic potential for the treatment of retinal degenerative diseases. Methods: The embryoid bodies were formed from hESCs and attached on the matrigel coated culture dishes. The neural structures consisting neural precursors were selected and expanded to form rosette structures. The mechanically isolated neural rosettes were differentiated into pigmented cells in the media comprised of N2 and B27. Expression profiles of markers related to RPE development were analyzed by reverse transcription-polymerase chain reaction and immunostaining. Dissociated putative RPE cells ($10^5$ cells/5 ${\mu}L$) were transplanted into the subretinal space of rat retinal degeneration model induced by intravenous sodium iodate injection. Animals were sacrificed at 1, 2, and 4 weeks after transplantation, and immnohistochemistry study was performed to verify the survival of the transplanted cells. Results: The putative RPE cells derived from hESC showed characteristics of the human RPE cells morphologically and expressed molecular markers and associated with RPE fate. Grafted RPE cells were found to survive in the subretinal space up to 4 weeks after transplantation, and the expression of RPE markers was confirmed with immunohistochemistry. Conclusion: Transplanted RPE cells derived from hESC in the defined culture condition successfully survived and migrated within subretinal space of rat retinal degeneration model. These results support the feasibility of the hESC derived RPE cells for cell-based therapies for retinal degenerative disease.

• Journal of Ginseng Research
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• v.47 no.2
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• pp.311-318
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• 2023

Background: The beneficial effects of compound K (CK) on different chronic diseases have been shown to be at least related to antioxidant action. Nevertheless, since its antioxidant activity in human retinal pigment epithelial (RPE) cells is still unknown, here we investigated whether CK alleviates oxidative stress-stimulated damage in RPE ARPE-19 cells. Methods: The cytoprotective consequence of CK in hydrogen peroxide (H₂O₂)-treated cells was evaluated by cell viability, DNA damage, and apoptosis assays. Fluorescence analysis and immunoblotting were performed to investigate the inhibitory action of CK on reactive oxygen species (ROS) production and mitochondrial dysfunction. Results: H₂O₂-promoted cytotoxicity, oxidative stress, DNA damage, mitochondrial impairment, and apoptosis were significantly attenuated by CK in ARPE-19 cells. Furthermore, nuclear factor erythroid 2-related factor 2 (Nrf2) phosphorylation level and its shuttling to the nucleus were increased, which was correlated with upregulated activation of heme oxygenase-1 (HO-1). However, zinc protoporphyrin, a blocker of HO-1, significantly abrogated the preventive action of CK in H₂O₂-treated ARPE-19 cells. Conclusion: This study indicates that activation of Nrf2/HO-1 signaling by CK plays an important role in rescuing ARPE-19 cells from oxidative cellular damage.

• Polymer(Korea)
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• v.38 no.3
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• pp.364-370
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• 2014

The retinal pigment epithelium (RPE) plays an important role in maintaining a healthy retina and the degeneration of RPE caused a number of retinal diseases. The transplantation of RPE has recently become a possible therapeutic modality for retinal degeneration. To transplant RPE cells securely, substrates are essential, and then as a substrate, we fabricated films using silk that has unique mechanical properties and biocompatibility. After the FTIR spectra, contact angle and biodegradation of silk films were confirmed, RPE cells were seeded and the influence of RPE cells on silk films was examined. We measured the cell adhesion, cell viability, morphology and specific mRNA expression by MTT assay, SEM, immunofluorescence and RT-PCR. In this study, we confirmed that attachment, proliferation and phenotype maintenance of RPE cells cultured on silk films were great, and thereby we were able to confirm the potential applications of silk films as tissue engineering carrier for regeneration of retina.

• Archives of Pharmacal Research
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• v.24 no.4
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• pp.349-354
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• 2001

Curcumin, a dietary pigment in turmeric, posseses anti-carcinogenic and anti-metastatic properties. The present study was conducted to study in vitro chemopreventive effects of curcumin in transformed breast cells. Here, we show that curcumin inhibits H-ras-induced invasive phenotype in MCF10A human breast epithelial cells (H-ras MCF10A) and downregulates matrix metalloproteinase (MMP)-2 dose-dependently. Curcumin exerted cytotoxic effect on H-ras MCF10A cells in a concentration-dependent manner. Curcumin-induced cell death was mainly due to apoptosis in which a prominent downregulation of Bcl-2 and upregulation of Bax were involved. We also suggest a possible involvement of caspase-3 in curcumin-induced apoptosis. Curcumin treatment resulted in the production of reactive oxygen species (ROS) in H-ras MCF10A cells. Apoptotic event by curcumin was significantly inhibited by pretreatment of an antioxidant N-acetyl-$_L$-cysteine (NAC), suggesting redox signaling as a mechanism responsible for curcumin-induced apoptosis in H-ras MCF10A cells. Taken together, our results demonstrate that curcumin inhibits invasion and induces apoptosis, proving the chemopreventive potential of curcumin .

• Applied Microscopy
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• v.27 no.1
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• pp.1-12
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• 1997

Dorsal and ventral epithelium of leech, Whitemenia edentula, were observed, using both light and electron-microscope. Results are as follows. Epithelia are composed of simple columnar or irregularly-shaped epithelium, the transveral folds were seen in the cross-sectioned dorsal and ventral epithelia, but only longitudinal folds(depth $500{\mu}m$, width $350{\mu}m$) were discovered in the ventral epithelium. As to muscular tissue under the dorsal epithelium of Whitemenia edentula, that of longitudinal muscle layer is thicker and more developed than that of circular muscle layer. Seven gland cells (type-A, B, C, D, E, F and H cells) and one pigment cell (type-G cell) were discovered in the whitemenia edentula. Those were identified as three kinds of the neutral mucopolysaccharide granoles (type-A, B and F cells), two kinds of acidic mucopolysaccharide granules (type-C and D cells), and two kinds of non-reactive granules respectively (type-E and F cells). The glanules of the type-A and type-B gland cells ae electronly high-dense, and surrounded by the muscular tissue. As to the distribution of gland cells, the type-A, type-B, type-C, type-D, type-I and type-H gland cells were discovered only between the connective tissue and the circular muscle layer, while type-F gland cells were discovered only between tile circular muscle layer and the longitudinal muscle layer.

• Environmental Analysis Health and Toxicology
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• v.21 no.1 s.52
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• pp.35-43
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• 2006

Reproduction and organ structure of the pale chub, Zacco platypus in Isa stream were investigated by means of histological methods. The results of the study confirmed reproductive abnormality and histopathological features in the pale chub. The gonadosomatic index (GSI) of the fish showed two peak in April and August. In summer season, GSI of the male was about two times of the female's GSI. Monthly variation of the gonadal development was very irregular. From the histological analysis of the organ structure, epidermal atrophy, necrosis and hyperplasia of pigment cell were observed in the skin. Epithelial layer lifting and clubbing of the lamella and bifurcation of the filament were observed in the gill. Also histological changes as congestion, cytoplasmic degeneration of hepatic cell, degeneration of bile duct, glomerular dilatation, degeneration of renal tubule and pycnosis of interstitial cell were identified in the liver and kidney, respectively.

• Applied Microscopy
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• v.20 no.1
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• pp.105-119
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• 1990

In order to know the morphological characteristics and changes according to the developmental stage, the comparative observations have been studied by light and electron microscope on the midgut(gastric caecum, stomach) of the grasshopper(Oxya sinuosa Mistshenko). The results obtained are summarized as follow : In light microscopic level, the cuboidal shaped cells of the gastric caecum in the 5th instar nymph are differentiated the columnar epithelium in the adult. A number of pigment granules are appeared in the gastric caecum of the 5th instar nymph, however the pigment granules were absent in the adult. Indistinct or undifferentiated folds of the epithelial layer were appeared in the 5th instar nymph, whereas the well-developed folds were appeared the gastric caecum in the adult. The well-developed muscular layers are seen in the 5th instar numph, however in the adult the muscular layers are appeared thin or a few layers. In electron microscopic level, in the midgut epithelium, a number of well-developed rER, a few lipid droplets, multi vesicular bodies, small vesicles and glycogen granules were found. Two types of the pigment granules in the gastric caecum of the 5th instar nymph were appeared, whereas the whorl-membrane, a few secretory granules and one type of the gastro-entero-endocrine cell were found in the adult midgut. The light and dark cells could be distinguished in the stomach epithelim of the 5th instar nymph.

• Applied Microscopy
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• v.27 no.2
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• pp.131-144
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• 1997

The present study was carried out to investigate the processes of the ultrastructural differentiation and the acetylcholinesterase (AChE) activities of the developing rat retina. The results are as follows. The retina of fetal rat on the 13th day of gestation showed the early stage of differentiation. Briefly, there appeared dividing chromosomes, the plentiful free ribosomes, and the high ratio of nucleus to cytoplasm. The reaction products by AChE were localized at the membrane of endoplasmic reticulum and on the outer membrane of nucleus. Ultrastructures and AChE activities in the retina of the fetal rats on the 18th day of gestation were similar to those of the prior stages, except the appearence of rough endoplasmic reticulum and Golgi apparatus. According to the ultrastructural observations, the rat retina was still in immature state at birth, but the pigment epithelial cells were fully differentiated, e. g. the increase of melanin granules, the development of mitochondria and Golgi apparatus. The AChE activity was weekly detected. The differentiated retinal layers and the outer segment of photoreceptor cells were observed on the 7th postnatal day. And the pigment epithelium appeared to be fully differentiated. On the 14th postnatal day, rat retina were completely differentiated. In other words, the rat retina was characterized by the prominent outer segments, phagocytosed residues in the pigment epithelium, and the localization of reaction products by AChE in the synapses. In conclusion, the differentiation of rat retina is charaterized by the changes of cell shape, the increase of retinal layers, and the alterations of AChE activities. It seems that rat retina is to be functional from 2 weeks of birth onward, coinciding with the eye opening of the juvenile rats.

• Molecules and Cells
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• v.46 no.7
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• pp.420-429
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• 2023

Age-related macular degeneration (AMD) is one of the leading causes of blindness in elderly individuals. However, the currently used intravitreal injections of anti-vascular endothelial growth factor are invasive, and repetitive injections are also accompanied by a risk of intraocular infection. The pathogenic mechanism of AMD is still not completely understood, but a multifactorial mechanism that combines genetic predisposition and environmental factors, including cellular senescence, has been suggested. Cellular senescence refers to the accumulation of cells that stop dividing due to the presence of free radicals and DNA damage. Characteristics of senescent cells include nuclear hypertrophy, increased levels of cell cycle inhibitors such as p16 and p21, and resistance to apoptosis. Senolytic drugs remove senescent cells by targeting the main characteristics of these cells. One of the senolytic drugs, ABT-263, which inhibits the antiapoptotic functions of Bcl-2 and Bcl-xL, may be a new treatment for AMD patients because it targets senescent retinal pigment epithelium (RPE) cells. We proved that it selectively kills doxorubicin (Dox)-induced senescent ARPE-19 cells by activating apoptosis. By removing senescent cells, the expression of inflammatory cytokines was reduced, and the proliferation of the remaining cells was increased. When ABT-263 was orally administered to the mouse model of senescent RPE cells induced by Dox, we confirmed that senescent RPE cells were selectively removed and retinal degeneration was alleviated. Therefore, we suggest that ABT-263, which removes senescent RPE cells through its senolytic effect, has the potential to be the first orally administered senolytic drug for the treatment of AMD.

• Applied Microscopy
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• v.32 no.3
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• pp.275-284
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• 2002

Granular gland regeneration in the toad after dorsal skin wound histologically was examined using scanning and transmission electron microscopy. After cutaneous wounds were induced by excision, animals were maintained in special cages for up to 20 days. In transmission electron microscopy (TEM), newly formed granular gland, though poorly developed, was seen on 4 day after injury. Epithelial cells moved toward apical region of newly formed gland. The cells had smooth surface and were not connected to other cells by desmosomes. Mitochondria rich cell (MRC) possessing long cytoplasmic processes formed a gland cavity and hemidesmosomes were found under the cell processes. Basal cavity of newly formed gland consisted of MRC, pro-granular producing cells (pGPC), and granular producing cell (GPC). Moreover it was observed that xanthophores moved to the base of the epithelial tissue on 10 day after the injury. These cells contained numerous pterinosomes and carotenoid vesicles. Immature pterinosomes were large and carotenoid vesicles were moderately electron dense. On 13 day after the injury, xanthophores contained abundant carotinoid vesicles and lammelated pterinosomes. Iridophores were also observed adjacent the developing xanthophores on 16 day post-injury. These observations indicated that regeneration of granular gland from glandular precursor cells during wound healing and subsequent expansion of the glandular cells might be dependent on maturation and proliferation of these newly formed cells.