• 동의신경정신과학회지
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• 제21권2호
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• pp.125-140
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• 2010

Objectives : The purpose of this study is to examine from various angles the protective effect of Gastrodia elata Blume (GEB) against nerve cell death induced by $\beta$-amyloid by using the cell line SH-SY5Y, which is commonly utilized for toxicity testing in nerve cells, and to find out its mechanism of action. Methods : To begin with, as a result of assessing the rate of cell survival by employing MTT reduction assay, the treatment with $\beta$-amyloid at different concentrations caused cytotoxicity, which was inhibited by preprocessing GEB extract. In addition, after $\beta$-amyloid was processed with the cell SH-SY5Y, apoptosis progressed, which was reduced effectively by processing GEB extract. Results : When cytotoxicity was caused by using hydrogen peroxide, a representative ROS, in order to examine the antioxidant effect of GEB, its protective effect was also observed. Apart from ROS, reactive nitrogen species (RNS) are also known to play a crucial role in nerve cell death. The treatment with the NO donor SNAP increased the production of nitric oxide and the expression of iNOS, which was also inhibited by GEB extract. Meanwhile, as an attempt to find out the mechanism of action explaining the antioxidant effect, the intracellular antioxidant enzyme expressions were measured by RT-PCR, which showed that GEB extract increased the expressions of heme oxygenase-1, GAPDH and $\gamma$-glutamate cysteine ligase. Lastly, GEB extract had a protective effect against impaired memory induced by scopolamine in animal models (in vivo). Conclusions : These findings indicate that GEB has a protective effect against the death of cranial nerve cells, suggesting possibilities for the prevention and treatment of AD.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제27권5호
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• pp.446-456
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• 2005

Nerve growth factor(NGF) has a critical role in peripheral nerve regeneration. The aim of this study is to construct a well-functioning hNGF-$\beta$ recombinat adenovirus for the ultimate development of improved method to promote peripheral nerve regeneration with adenovirus mediated hNGF-$\beta$ gene transfection into Schwann cells. First PCR associated cloning of GFP-tagged hNGF-$\beta$ which was ligated into E1/E3 deleted adenoviral vector was performed and tranfected into E. coli to construct hNGF-$\beta$ recombinant adenovirus. After production of recombinat adenovirus in a large scale, its transfection efficiency, expression, and function were evaluated using cell lines or primarily cultured cells of HEK293 cells, Schwann cells, fibroblast(NIH3T3) and myocyte(CRH cells). GFP expression was observed in 90% of infected cells compared to uninfected cells. Total mRNA isolated from hNGF-$\beta$ recombinat adenoviru infected cells showed strong RT-PCR band, however, LacZ recombinant adenovirus infected or uninfected cells did not. NGF quantification by ELISA showed a maximal release of 18.865 +/- 0.31ng/mL at 4th day. PC-12 cells exposed to media with hNGF-$\beta$ recombinant adenovirus infected Schwann cell demonstrated higher levels of differentiation compared with controls. We generated hNGF-$\beta$ recombinant adenovirus and induced over expression of NGF successfully in nonneuronal and neuronal cells. Following these result, it is expected to develop an improved treatment strategy peripheral nerve regeneration using the hNGF-$\beta$ gene transfected cells.

• Applied Microscopy
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• 제32권3호
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• pp.247-255
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• 2002

두족류 Octopus minor와 Todarodes pacificus의 시엽내 신경전달물질을 분비하는 neuron의 특성 및 기능을 확인하기 위해 serotonin 및 somatostatin의 항체를 사용한 면역염색과 면역금표지법을 시행하였다. 면역염색 결과 항-somatostatin은 살오징어인 경우 항-serotonin 면역반응과 유사하게 나타났지만, 서해낙지에서는 외과립세포층의 큰 세포에서만 반응을 보였다. 항-serotonin을 이용한 면역금표지법은 살오징어인 경우 내과립세포층과 수질부의 신경세포에서는 세포질 $0.5{\mu}m^2$당 30개 정도의 금입자가 관찰되어 강한 반응을 보인 반면, 서해낙지에서는 17개 정도의 비교적 약한 반응을 보였다. 항-somatostatin에서는 살오징어의 외과립 및 내과립세포층 그리고 수질부의 반응된 세포의 세포질 $0.5{\mu}m^2$당 30개 정도의 금입자가 관찰되어 강한 반응을 보인 반면, 서해낙지의 외과립세포층의 세포에서는 3개 정도의 금입자만이 관찰되어 역시 약한 반응을 보였다. 이와 같이 시엽의 각 부위별 면역염색과 면역금표지법을 시행한 결과 2종류의 항체에 각각 양성반응을 보인 신경세포들이 두 종에서 다양하게 분포하고 있음을 확인했는데 특히 면역염색과 면역금표지법에 관한 반응외 정도를 비교하면 서해낙지에 비해 살오징어에서 비교적 강하게 나타났다.

• 폴리머
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• 제32권1호
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• pp.49-55
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• 2008

슈반세포는 강력한 말초신경 재생 효과를 가지고 있다. 그리고 신경 유도관으로 사용된 소장점막하조직(small intestinal submucosa, SIS)은 세포활성을 촉진하는 여러 가지 성장인자를 다량 함유하고 있고 이종이식의 면역 거부반응이 없는 생체물질로서 널리 사용되고 있다. 슈반세포는 Fischer 쥐의 좌골신경으로부터 분리하였고, 순수 배양된 슈반세포를 $2{\times}10^6$ 세포로 계산하여 SIS 스폰지에 파종하였다. 그리고 Fischer 쥐의 좌골신경 5 mm를 제거하여 신경과 제조된 스폰지를 연결하는 신경 외막 봉합을 실시하였다. 그런 후 1, 2주 및 4주 후 재생된 신경을 적출 한 후, H &E 염색과 S-100, GFAP 및 NF 면역조직화학염색을 실시하였다. 그 결과, SIS 스폰지와 슈반세포를 함께 이식한 군(Group II)은 아무것도 이식하지 않은 군(Control II)과 SIS 스폰지만 이식한 군(Group I) 보다 뚜렷한 양성반응을 보였고 손상을 받지 않은 군(Control I)과 별차이가 없는 양상을 보여주었다. 이는 SIS 스폰지와 슈반세포를 함께 이식했을 때 손상된 조직공학적 말초 신경 재생에 중요한 역할을 한 것으로 사료된다.

• 대한한의학회지
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• 제18권1호
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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.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제32권4호
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• pp.295-307
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• 2006

Styela clava, called non-native tunicate or sea squirt, is habitat which include bays and harbors in Korea and several sites in the sea faced world. We fabricate cellulose membrane nerve conduit (CMNC) from this native sea squirt skin, and evaluate the capacity of promoting peripheral nerve regeneration in the rat sciatic nerve defect model. After processing the pure cellulose membrane from the sea squirt skin as we already published before, CMNC was designed as a non-tubular sheet with 14 mm length and 4 mm width. Total eleven male Spraque-Dawley rats (12 weeks, weighing 250 to 300g) were divided into sham group (n=2), silicone tube grafted control group (n=3) and experimental group (n=6). Each CMNC grafted nerve was evaluated after 4, 8 and 12 weeks in the experimental group, and after 12 weeks, sciatic function was evaluated with sciatic function index (SFI) and gait analysis, and histomorphology of nerve conduit and the innervated tissues of sciatic nerve were all examined using image analyzer and electromicroscopic methods in the all groups. The regenerated axon and nerve sheath were found only in the inner surface of the CMNC after 4 weeks and became more thicker after 8 and 12 weeks. In the TEM study, CMNC grafted group showed more abundant organized myelinated nerve fibers with thickened extracellular matrix than silicone conduit grafted group after 12 weeks. The sciatic function index (SFI) and ankle stance angle (ASA) in the functional evaluation were $-47.2{\pm}3.9$, $35.5^{\circ}{\pm}4.9^{\circ}$ in CMNC grafted group (n=2) and $-80.4{\pm}7.4$, $29.2^{\circ}{\pm}5.3^{\circ}$ in silicone conduit grafted group (n=3), respectively. And the myelinated axon was 41.59% in CMNC group and 9.51% in silicone conduit group to the sham group. The development of a bioactive CMNC to replace autogenous nerve grafts offers a potential and available approach to improved peripheral nerve regeneration. As we already published before, small peptide fragment derived from the basement membrane matrix proteins of squirt skin, which is a kind of anchoring protein composed of glycocalyx, induced the effective axonal regeneration with rapid growth of Schwann cells beneath the inner surface of CMNC. So the possibilities of clinical application as a peripheral nerve regeneration will be able to be suggested.

• Biomolecules & Therapeutics
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• 제17권4호
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• pp.362-369
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• 2009

We have investigated the effect of glucosamine on the retinal cells after continuous infusion into cerebroventricle by using osmotic minipump to avoid peripheral effect. Continuous intracerebroventricular (i.c.v) infusion of glucosamine with the rate of 0.1 ${\mu}mol$/10 ${\mu}l$/hr for 7 days resulted in morphological changes of the optic nerve in electron microscopic level as well as morphological changes of the retina in light microscopic level. Retinal sections were immunostained for the detection of morphological changes of astrocytes. GFAP immunoreactivity appeared not only in the Muller cells but also many of the radial processes of Muller cells. The optic nerve showed deformed axon and slight lamellar separation of myelin sheath after continuous infusion of glucosamine in observing with electron microscope. Interestingly, vacuoles were observed in deformed axons and retinal layers were folded and detached. These results suggested that glucosamine plays a role in induction of morphological dysfunction in retina and optic nerves.

• Journal of Chest Surgery
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• 제30권10호
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• pp.1051-1053
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• 1997

악성신경초종은 Schwan cell 또는 신경초 세포(nerve sheath cell)에서 발생하는 아주 드문 종양으로 빈번히 신경섬유종증(Von Rechlinghausen's disease)과 연관되어 있다. 환자는 64세 남자로 우측 가슴에 내원 2개월전부터 있던 무통성 종물을 주소로 내원하였으며 신경섬유종 증과의 동반은 없었다 전산화 단층촬영상 종물의 크기는 6 $\times$ 6 cm였으며, 11번 늑골과 횡경막을 침범하였 고 우신을 앞쪽으로 밀고 있었다. 본 교실에서는 흉벽에 발생한 악성신경초종을 경험하였기에 문헌고찰과 함께 보고한다.

• 대한수의학회지
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• 제54권1호
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• pp.7-12
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• 2014

Peripheral nerve sheath tumors (PNSTs) are heterogeneous tumor groups of peripheral nerves that originate from either Schwann cells or modified Schwann cells, fibroblasts, or perineural cells. In this study, signalment and clinical data such as tumor location and size were evaluated for 15 cases of PNSTs collected from local animal hospitals. The mean age of dogs with malignant PNST was higher than that of dogs with benign PNST. Additionally, the male to female ratio in dogs with PNST was 1 : 4. In dogs with PNST, the primary sites of involvement were the hindlimb, forelimb, around the mammary glands, the neck, and the abdomen. Histiopathologic examination revealed that eight PNSTs were benign and seven were malignant. The tumor cells were composed of loosely to densely arranged interlacing bundles and wavy spindle cells arranged in short bundles, palisading, and whirling. High mitotic figures, local invasion, multifocal necrosis and atypical multinucleated giant cells were observed in malignant PNST cases. All PNSTs showed immunoreactivity for vimentin and S-100. However, only 93.3% and 73.3% were immunoreactive for NSE and GFAP, respectively. Overall, these results indicated that immunohistochemical markers such as vimentin, S-100 and NSE could help confirm the diagnosis of canine PNSTs.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제30권6호
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• pp.465-473
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• 2004

Purpose : The essential triad for nerve regeneration is nerve conduit, supporting cell and neurotrophic factor. In order to improve the peripheral nerve regeneration, we used polyglycolic acid(PGA) tube and brain-derived neurotrophic factor(BDNF) gene transfected Schwann cells in sciatic nerve defects of SD rat. Materials and methods : Nerve conduits were made with PGA sheet and outer surface was coated with poly(lactic-co-glycolic acid) for mechanical strength and control the resorption rate. The diameter of conduit was 1.8mm and the length was 17mm Schwann cells were harvested from dorsal root ganglion(DRG) of SD rat aged 1 day. Schwann cells were cultured on the PGA sheet to test the biocompatibility adhesion of Schwann cell. Human BDNF gene was obtained from cDNA library and amplified using PCR. BDNF gene was inserted into E1 deleted region of adenovirus shuttle vector, pAACCMVpARS. BDNF-adenovirus was multiplied in 293 cells and purified. The BDNF-Adenovirus was then infected to the cultured Schwann cells. Left sciatic nerve of SD rat (250g weighing) was exposed and 14mm defects were made. After bridging the defect with PGA conduit, culture medium(MEM), Schwann cells or BDNF-Adenovirus infected Schwann cells were injected into the lumen of conduit, respectively. 12 weeks after operation, gait analysis for sciatic function index, electrophysiology and histomorphometry was performed. Results : Cultured Schwann cells were well adhered to PGA sheet. Sciatic index of BDNF transfected group was $-53.66{\pm}13.43$ which was the best among three groups. The threshold of compound action potential was between 800 to $1000{\mu}A$ in experimental groups which is about 10 times higher than normal sciatic nerve. Conduction velocity and peak voltage of action potential of BDNF group was the highest among experimental groups. The myelin thickness and axonal density of BDNF group was significantly greater than the other groups. Conclusion : BDNF gene transfected Schwann cells could regenerate the sciatic nerve gap(14mm) of rat successfully.