• Maxillofacial Plastic and Reconstructive Surgery
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• v.27 no.5
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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.

• Journal of Oral Medicine and Pain
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• v.24 no.2
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• pp.145-161
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• 1999

The proto-oncogene bcl-2 confers a survival advantage to cells by blocking programmed cell death (apoptosis). Overexpression of bcl-2 probably plays a role in tumorigenesis, and the expression of the bcl-2 protein has been investigated in many kinds of tumors. An increased expression of nitric oxide synthetase(NOS) has been observed in human colon cancer cell lines as well as in human gynecological, breast, and CNS tumors. However there have been only a few reports on the expression of bcl-2 and $NOS_2$ in oral white lesions and cancer. The aim of this study was to investigate the relationship between the expression of Bcl-2 and $NOS_2$ and several pathological parameters such as histological types and layers. We reported desregulation of bcl-2 and $NOS_2$ expression during progression from oral white lesion, lichen planus and leukoplakia to squamous cell carcinoma. The obtained results were as follows: 1. Immunohistochemical analysis with monoclonal antibodies to bcl-2 oncoprotein and $NOS_2$ in formalin-fixed paraffin-embedded tissue sections revealed that bcl-2 expression is restricted to the basal cell layer and $NOS_2$ was mild expressed only in subepithelial inflammatory cells in normal human mucosa. There wasn't specific finding of those in lichen planus and leukoplakia. 2. Bcl-2 immunoreactivity in severe epithelial dysplasia or CIS occurs throughout the epithelium, $NOS_2$ reactivity in most superficial layer were noted. 3. In well-differentiated squamous cell carcinomas, mostly bcl-2 was overexpressed. In moderated and poor squamous cell carcinomas, the expression of $NOS_2$ was increased and that of bcl-2 was decreased. 4. The immunoreactivity of bcl-2 was 12.5% of normal mucosa, 30% of leukoplakia, 44% of lichen planus and 67% of carcinoma in situ. In carcinoma, those were 43%, 50% and 67% according to differentiation, respectively. 5. The immunoreactivity of $NOS_2$ was 25% of normal mucosa, 70% of leukoplakia, 78% of lichen planus and 100% of carcinoma in situ and epithelial dysplasia. In carcinoma, those were higher in moderated(100%) and poor(83%) squamous cell carcinomas than in well differentiated type(71%). 6. The expression of bcl-2 and $NOS_2$ by Western blot was increased highly in lichen planus and leukoplakia. Therefore, the expression of bcl-2 was increased in the white and precancerous lesions and that was decreased by differentiation of carcinoma. However, $NOS_2$ immunoreactivity in carcinoma in situ was lower than those in moderated and poor squamous cell. These findings suggest that the interaction of bcl-2 and $NOS_2$ may be roled importantly in growth and development of carcinoma.

• The Journal of Korean Academy of Prosthodontics
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• v.43 no.4
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• pp.511-518
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• 2005

Purpose: The purpose of this study is to find the effect of rare earth magnet's magnetic field of to the osteoblast around the implant by the means of observation number, and distribution around the implant which is connected to the permanent magnet but not, counted and compared by the number of cells attached to the surface of the implant. Material and method: The permanent magnets, made in the healing cap form, were connected to the implant future, and placed on the culture plate, The osteoblast-like cell: MC3T3-E1 were used for cell culture. As the control group, the implant were connected to normal healing cap, and cultured in the same conditions. 48 hours later, using inverted microscope, the number and distribution of osteoblast around the implant were observed, and 72 hours later, the number of the cells attached to the implant were counted. Results: As a result, the implant connected to the permanent magnet had proved to have a more concentrated cell distribution rate than the control group. The implant connected to the permanent magnet, neck area : which has about 10 gauss magnetic force, had more cells than apex area. The implant connected to the permanent magnet had proven to attach to the osteoblast more productively than control group's implant. Conclusions: This research showed that the magnetic field of the permanent magnet affected the distribution and growth rate of the osteoblast around the implant. In order to support this study, it also had need to monitor the progress of the permanent magnet specifically shown on the neck area, which has10 gauss magnetic force. So after additional research on the distribution and attachment of the cells, and further more, on bone formation, it will be concluded that the clinical applications ,such as immediate loading of implant treatment are possible.

• KSBB Journal
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• v.19 no.1
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• pp.17-26
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• 2004

In this study the extracellular production of 5-aminolevulinic aicd (ALA) by recombinant E. coli BL2l (DE3) pLysS harboring the plasmid pFLS45 are investigated. Optimum concentrations of succinic acid and glycine for cell growth and ALA production were found to be 30 mM and 15 mM, respectively. Levulinic acid (LA) as an inhibitor of ALAD was added to the culture medium in the end of exponential cell growth phase and its optimum concentration was 30 mM. Growth of recombinant E. coli BL2l (DE3) pLysS (pFLS45) was largely dependent upon the pH value of culture medium. When the pH of culture medium was in the range of 6.0 and 6.5, high cell mass and ALA production were obtained. IPTG induction for the expression of the fusion gene did not enhance the production of ALA. Recombinant cell grew at 30't faster than at 37$^{\circ}C$, but ALA productivity was lower than at 37$^{\circ}C$. Repeated addition of glycine, succinic acid, and LA increased the production of ALA and the inhibition of intracellular ALA dehydratase activity, with up to 1.3 g/L ALA having been produced in the cultivation.

• Korean Journal of Animal Reproduction
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• v.24 no.4
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• pp.439-449
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• 2000

This study was to investigate the expression of transgene after co-injection of spermatozoon and EGFP gene into mature oocytes in cattle. From frozen semen, spermatozoa were treated by DTT with 0.03% Tween-20, freezing and thawing or 0.02% Triton X-100 to disrupt their plasma membranes. The sperm injected oocytes were co-cultured with bovine oviduct epithelial cells in CRlaa, and expression of EGFP in embryos were observbed under epifluorescent microscope. Two pronuclei were formed in oocytes injected with sperm treated by DTT (44.6%), DTT-Tween-20 (48.4%), DTT-freezing (44.4%) and DTT-Triton X-100 (42.9%). Cleavage and blastocyst formation rates of bovine oocytes which injected with sperm treated by DTT, DTT-Tween-20, DTT-freezing, and DTT-Triton X-100 were 49.1, 58.5, 43.9, and 48.4% and 10.2, 13.0, 6.8, and 6.5%, respectively. Although the most of embryos were showing mosaicism, embryos expressing EGFP gene were observed in all treated groups. Therefore, these results indicate that membrane disrupted sperm could interact with exogenous DNA, and that this procedure may be useful to introduce foreign gene into bovine oocytes.

• Journal of Animal Science and Technology
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• v.44 no.6
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• pp.677-684
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• 2002

We have investigated the expression patterns of candidates for growth stage specifically expressed genes. The expression patterns of the EPV20, aldolase A, Translationally Controlled Tumor Protein (TCTP) and Adipocyte Differentiation Related Protein (ADRP) were examined by semiquantitative RT-PCR and northern blot analysis in skeletal muscle tissues of Hanwoo, especially in the longissimus dorsi at various growth stages. The EPV20 mRNA was expressed in longissimus dorsi tissue of Hanwoo, but there was no difference of expression levels during growth stages. Though the aldolase A gene was reported to be muscle-specific and regulated at developmental stages, the expression levels of aldolase A mRNA in the longissimus dorsi tissues showed little differences at various growth stages. The expression levels of TCTP which was reported as growth-related protein regulated at translation step were gradually increased during growth of Hanwoo. The expression levels of ADRP mRNA were rapidly increased at 24-month-old longissimus dorsi tissue of Hanwoo, and decreased at 30-month-old. Our data suggest that the ADRP gene show as growth-stage dependent expression and is related to fat deposition within muscular tissue.

• Research in Plant Disease
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• v.19 no.3
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• pp.226-228
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• 2013

In 2012, a pink mold rot was observed on unishiu orange (Citrus unshiu Mac.) fruits at the Wholesale Market for Agricultural Products, Jinju, Korea. The symptom on unishiu orange was a water-soaked lesion on the surface of fruit, which later on enlarged to form softened brown rot lesions. The diseased fruits were covered with pink-colored mold, consisting of conidia and conidiophores of the pathogen. Optimum temperature for mycelial growth was $25^{\circ}C$. Conidia were hyaline, smooth, 2-celled, and thick-walled conidia with truncate bases, ellipsoidal to pyriform, characteristically held together zig-zag chains and $12-26{\times}8-12{\mu}m$ in size. Conidiophore was erect, colorless, unbranched, and 4-5 ${\mu}m$ wide. On the basis of mycological characteristics, pathogenicity test, and molecular analysis with complete ITS rDNA region, the causal fungus was identified as Trichothecium roseum (Pers.) Link ex Gray. This is the first report of pink mold rot caused by T. roseum on unishiu orange in Korea.

• Applied Microscopy
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• v.27 no.2
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• pp.121-130
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• 1997

It has been demonstrated that majority of cells in the mammalian body such as myocytes and epithelial cells of skin and intestine respond to mechanical force or environmental factors and exhibit partial disruption of cell membrane, i. e., cell wounding, even in a physiological condition. Myocardial cells are rather apt to be wounded than other cells since they are definitely exposed to mechanical stress by contraction-relaxation and blood flow. However, the mechanism how myocardial cells protect themselves against cell wounding is not yet clarified. On this background, the present study was performed to elucidate whether albumin leakage is related to cell wounding and to assess whether diltiazem, a potent calcium channel blocker, is beneficial in isoproterenol-induced cell wounding in the heart. Hearts isolated from New Zealand White rabbits ($1.5\sim2.0kg$ body weight, n=20) were perfused with Tyrode solution by Langendorff technique. After stabilization of baseline hemodynamics, the hearts were subjected to bolus administration of isoproterenol and diltiazem as following order: $1.6{\mu}M$ isoproterenol at zero min (the beginning point): $16{\mu}M$ diltiazem at 20min; $1.6{\mu}M$ isoproterenol at 25min; $16{\mu}M$ isoproterenol at 45 min; $160{\mu}M$ diltiazem at 65 min; $16{\mu}M$ isoproterenol at 70 min. During all experiments, the left ventricular function was recorded, albumin leakage in the coronary effluents was analyzed by electrophoresis and Western blot, and myocardial cell membranes were examined by conventional transmission electron microscopy. Data were analyzed by t-test and linear regression test. Isoproterenol significantly increased the inotropic and chronotropic contractions, coronary flow, and frequency of arrhythmia, however, diltiazem did not influence on hemodynamics except decrease in the frequency of arrhythmia and a slight decrease in contractility. Isoproterenol also resulted partial disruption of myocardial cell membrane and inclose in albumin leakage, while diltiazem pretreatment showed number of electron-dense plaques in the cell membrane and a tendency of decrease in albumin leakage. These results indicate that albumin leakage may be an indirect index of cell wounding in the heart and diltiazem nay be beneficial to protect myocardial cells against isoproterenol-induced cell wounding. It is likely that diltiazem promotes resealing process of the cell membrane.

• The Journal of Internal Korean Medicine
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• v.39 no.5
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• pp.973-983
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• 2018

Objectives: We report a case of traditional Korean medicine (TKM) treatment for skin side effects after taking afatinib (Giotrif$^{(R)}$). Methods: A 62-year-old female who was diagnosed with non-small cell lung cancer stage 4 (T4N2M1b) and was on treatment with afatinib (29.56 mg/day for 4 months) complained of skin toxicity as a side effect. For 16 admission days, the patient was treated with acupuncture, moxibustion, and herbal medicine (oral decoction and external ointment). Results: Improvement of skin toxicity was measured by a numeric rating scale. In addition, Quality of life (QOL) was measured using EORTC Quality of Life Questionnaire, Core 30 (EORTC QLQ-C30) and EORTC Quality of Life Questionnaire, 13-item lung cancer-specific module (EORTC QLQ-LC13) Developed by the European Organization for Research and Treatment of Cancer (EORTC). Tumor size and carcinoembryonic antigen (CEA) were also examined during follow up. Conclusions: After a combined TKM treatment, skin toxicity symptoms and quality of life scales were significantly improved with no side effects. The tumor size was not changed on computed tomography during follow-up period. CEA levels were decreased.

• Journal of Oriental Neuropsychiatry
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• v.20 no.2
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• pp.111-120
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• 2009

Objectives : This study was performed to investigate the effects of Needle Electrode Electrical Stimulation (NEES) on ischemia-induced cerebrovascular accidents. After obstruction and reperfusion of ${\ast}{\ast}$ arteries in white mice, the amounts of necrosis and inflammation related substances IL-6, Caspase-3, and PARP, C-fos were measured in neurons of the hippocampus. The following results were obtained. Methods : This study used 21 male specific pathogen free (SPF) SD (Sprague Dawley) rats, 8 weeks of age and approximately 300 g in weight, that were given at least 1 week to adapt to the lab environment Each exposed artery was completely occluded with non-absorbent suture thread and kept in that state for 5 minutes. The sutures were then removed to allow reperfusion of blood. Test group is control group for comparison with the common carotid artery occlusion models, a GI group that underwent common carotid artery occlusion, and a needle electrode electrical stimulation (NEES) group that underwent NEES after artery occlusion. The GI and NEES groups were given 12, 24, or 48 hours of reperfusion before NEES. NEES device (PG6, ITO, Japan, 9V, current, 2Hz) was used to stimulate the right and left acupoint ST36 of the SD rats for 30 minutes while they were sedated with 3% isoflurane. An immunohistochemistry test was done on the forebrains of the GI induced rats. All the data collected from this study was symbolized and analyzed using a statistics processing program (SPSS 12.0K/PC). The level of significance was set at ${\alpha}$=0.05 and a T-TEST analysis was used to find out the effects of treatment on each of the groups: the normal group, the CVA induced group, and the treatment after CVA induction group. Results : Both PARP and C-fos immuno-reactive cells, related to apoptosis, were greater in the GI groups than the NEES group. Caspase and IL-6 immuno-reactive cells, related to inflammation, were greater in the GI and NEES groups than the control group. Conclusions : This research was conducted to study the effects of NEES on CVA due to ischemia. Occlusion and reperfusion was performed on the common carotid arteries of white rats, after which amounts of substances related to neuron necrosis and inflammation - PARP, IL-6, Caspase-3, and C-fos - were measured in the Hippocampus