• Journal of Acupuncture Research
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• v.21 no.2
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• pp.1-20
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• 2004

Objective : The aim of this study was to investigate effects of Angelica gigas Nakai(AGN) on the ischemic injury by intraluminal filament insertion in the rats. Methods : The ischemia was induced by intraluminal filament insertion into middle cerebral artery. AGN herbal acupuncture into SP10 was carried out during 3 weeks after ischemic injury. Eight-arm radial maze was designed for the behavioral task. AGN herbal acupuncture showed neuroprotective agents in cresyl violet, acetylcholinesterase(AchE), choline acetyltransferase(ChAT) and nerve growth factor(NGF)-stain. Then check the effect of regional cerebral blood flow(rCBF) according to AGN herbal acupuncture in rats. Results : The errors in the eight-arm radial maze task were significantly decreased in normal group compared with control group on 1~6days, AGN2(0.02g/kg) herbal acupuncture group on 1~5days, AGN3(0.1g/kg) on 1~3days, AGN4(0.5g/kg) on 1, 3~6days. The rate of correct choice was significantly increased in AGN1(0.01g/kg) and AGN4 herbal acupuncture groups. The density of neurons in the hippocampal CA1 was the most increased in normal group and AGN1, AGN3, AGN4 herbal acupuncture groups compared with control group. The density of AchE in the hippocampal CA1 had a tendency to increase in all the groups when they were compared with control group, but not significant. The density of ChAT in the hippocampal CA1 was significantly increased in normal group and AGN1, AGN4 herbal acupuncture groups compared with control group. The density of NGF in the hippocampal CA1 was significantly increased AGN4 herbal acupuncture group compared with control group. The rCBF was significantly increased in AGN1, AGN3 and AGN4 herbal acupuncture groups without the change of blood pressure. Conclusions : These results suggest that AGN herbal acupuncture can be used for controlling stroke in early stage as herbal medication.

• Journal of Oriental Neuropsychiatry
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• v.18 no.1
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• pp.153-171
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• 2007

Objective : The aim of this study was to investigate effects of Gagambosim-Tang(GBT) of focal brain ischemic injury induced by intraluminal filament insertion in the rats. Method : The ischemia was induced by intraluminal filament insertion into middle cerebral artery Eight-arm radial maze was designed for the behavioral task. Gagambosim-Tang was orally administrated in SD rats for 21 days. The task was started on the 4th week after focal brain injury, and performed two trials per day for 6days. The effects of Gagambosim-Tang on neuroprotective agents in cresyl violet, choline acetyltranferase(ChAT), nerve growth factor (NGF)-stain and c-Fos with ischemic injury were investigated. Results : The errors in the eight-arm radial maze task were significantly decreased in normal group compared with control group on 2-6days, GBT lX(42.2 mg/ml)orally administrated group on 1days, GBT 6X(253.2 mg/ml) on 3, 5, 6days. The rate of correct choice was increased in normal and GBT 6X groups. The neuroprotective effect in the hippocampal CA1 was increased in normal and GBT 1X, GBT 6X groups compared with control group. The density of ChAT in the hippocampal CA1 was increased in normal and GBT6X groups compared with control group. The density of NGF in the hippocampal CA1 was increased normal and GBT6X groups compared with control group. The number of c-Fos-positive neurons in the hippocampal CA1 was increased in normal and GBT 6X groups compared with control group. Conclusion : These results suggest that Gagambosim-Tang may have protective effect on dementia.

• BMB Reports
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• v.43 no.5
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• pp.369-374
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• 2010

The PC12 is the widely used cell line to study neuronal differentiation. We had extensively investigated the details of protein expression in differentiated PC12 cells by proteomic analysis. The cells were incubated at the presence of nerve growth factor. We had analyzed the expression changes in the differentiating PC12 cells by 2-dimensional electrophoresis and the identification of the proteins using MALDI-TOF MS. By comparing expression pattern in the time course, we identified the candidate genes which are associated with neuronal differentiation. Among these genes, we performed real-time PCR analysis to validate $Idh3{\alpha}$ expression by the time course. To identify the function of $Idh3{\alpha}$ in neuronal differentiation stage, the transfection of $Idh3{\alpha}$ to PC12 cells was performed. As a result, we proved that up-regulation of $Idh3{\alpha}$ causes reduction in neural differentiation of PC12 cells. Based on these data, we suggest that $Idh3{\alpha}$ plays a role to the neuronal differentiation.

• Archives of Plastic Surgery
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• v.32 no.4
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• pp.408-415
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• 2005

Following a transverse rectus abdominis musculocutaneous(TRAM) flap breast reconstruction, denervated state of the flap causes the flap skin prone to thermal injury, calling for special attention. During the last 5 years, 69 breast reconstruction with 72 free TRAM flaps, were performed. Four out of thesse 69 patients sustained burn injury. Heat sources were a warm bag(n=2), heating pad(n=1) and warming light (n=1). The thermal injuries occured from 2 days to 3 months following the reconstruction. Three patients healed with conservative treatment, but one patient required debridement and skin graft. Initially 3 out of 4 patients with the burn had shown superficial 2nd degree burn with small blebs or bullae. However all 4 patients healed with scars. Mechanism of burn injuries of the denervated flap are known to be resulting from; 1) loss of behavioral protection due to denervation of flap with flap elevation and transfer, 2) loss of autonomic thermoregulatory control with heat dissipation on skin flap vasculature contributing to susceptibility of burn injury. 3) changes of immunologic and normal inflammatory response increasing thromboxane, and a fall in substance P & NGF (nerve growth factor). Including the abdominal flap donor site, sensory recovery of the reconstructed breast varies individually from 6 month even to 5 years postoperatively. During this period, wound healing is delayed, resulting in easier scarring compared to that observed in the sensate skin. Patients should be carefully informed and warned of possible burn injuries and taught to avoid exposure to heat source at least until 3 years postoperatively.

• Archives of Plastic Surgery
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• v.32 no.4
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• pp.403-407
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• 2005

Following a transverse rectus abdominis musculocutaneous(TRAM) flap breast reconstruction, denervated state of the flap causes the flap skin prone to thermal injury, calling for special attention. During the last 5 years, 69 breast reconstruction with 72 free TRAM flaps, were performed. Four out of thesse 69 patients sustained burn injury. Heat sources were a warm bag(n=2), heating pad(n=1) and warming light (n=1). The thermal injuries occured from 2 days to 3 months following the reconstruction. Three patients healed with conservative treatment, but one patient required debridement and skin graft. Initially 3 out of 4 patients with the burn had shown superficial 2nd degree burn with small blebs or bullae. However all 4 patients healed with scars. Mechanism of burn injuries of the denervated flap are known to be resulting from; 1) loss of behavioral protection due to denervation of flap with flap elevation and transfer, 2) loss of autonomic thermoregulatory control with heat dissipation on skin flap vasculature contributing to susceptibility of burn injury. 3) changes of immunologic and normal inflammatory response increasing thromboxane, and a fall in substance P & NGF (nerve growth factor). Including the abdominal flap donor site, sensory recovery of the reconstructed breast varies individually from 6 month even to 5 years postoperatively. During this period, wound healing is delayed, resulting in easier scarring compared to that observed in the sensate skin. Patients should be carefully informed and warned of possible burn injuries and taught to avoid exposure to heat source at least until 3 years postoperatively.

• Korean Journal of Pharmacognosy
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• v.48 no.4
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• pp.261-267
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• 2017

As part of the search for bioactive constituents of Korean medicinal plants, twelve steroids (1-12) were isolated from the rhizomes of Dioscorea nipponica. The isolated compounds were identified as diosgenin ($3{\beta}$, 25R)-spirost-5-en-3-ol (1), 25(R)-dracaenoside E (2), dioscin (3), gracillin (4), prosapogenin B (5), 25(R)-dracaenoside G (6), diosgenin 3-O-${\beta}$-D-glucopyranosyl($1{\rightarrow}3$)-${\beta}$-D-glucopyranoside (7), ophipogonin C′ (8), 7-oxodioscin (9), protodioscin (10), hypoglaucin F (11), and protoneogracillin (12). Their structures were characterized by spectroscopic data and identified by comparing these data with those in the literatures. All the isolates (1-12) were evaluated for their neuroprotective effects through induction of nerve growth factor in C6 glioma cells and effects on nitric oxide (NO) production in murine microglia cell line BV-2. Compounds 7 and 12 were found to induce upregulation of NGF secretion without causing significant cell toxicity and compound 4 exhibited potent anti-neuroinflammatory activity.

• Journal of Physiology & Pathology in Korean Medicine
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• v.17 no.2
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• pp.529-541
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• 2003

Yukmijihwang-tang has been widely used as an and-aging herbal medicine for hundred years in Asian countries. Numerous studies show that Yukmijihwangtang has anti-oxidative effect both in vivo and in vitro. It has been reported that Moutan Cortex Radicis extract (MCR) was the most effective herb in Yukmijihwang-tang on undifferentiated PC12 cells upon oxidative-stressed with hydrogen peroxide. The purpose of this study is to; 1) evaluate the recovery of neuronal damage by assessing the anti-oxidant effect of MCR on PC12 cells differentiated with nerve growth factor (NGF), 2) identify candidate genes responsible for anti-oxidative effect on differentiated PC12 cells by oligonucleotide chip microarray. PC12 cells, which were differentiated by treating with NGF, were treated without or with hydrogen peroxide in the presence or absence of various concentration of MCR. Cell survival was determined by using MTS assay. Measurement of intracellular reactive oxygen species (ROS) generation was determined using the H2DCFDA assay The viability of cells treated with MCR was significantly recovered from stressed PC12 cell. In addition, wide rage of concentrations of MCR shows dose-dependent inhibitory effect on ROS production in oxidative-stressed cells. Total RNAs of cells without treatment(Control group), only treated with H₂O₂ (stressed group) and treated with both H₂O₂ and of MCR (MCR group) were isolated, and cDNAs was synthesized using oligoT7(dT) primer. The fragmented cRNAs, synthesized from cDNAs, were applied to Affymetrix GeneChip Rat Neurobiology U34 Array. mRNA of Calcium/calmodulin-dependent protein kinase II delta subunit(CaMKII), neuron glucose transporter (GLUT3) and myelin/oligodendrocyte glycoprotein(MOG) were downregulated in Stressed group comparing to Control group. P2X2-5 receptor (P2X2R-5), P2X2-4 receptor (P2X2R-4), c-fos, 25 kDa synaptosomal attachment protein(SNAP-25a) and GLUT3 were downregulated, whereas A2 adenosine receptor (A2AR), cathechol-O-methyltransferase(COMT), glucose transporter 1 (GLUT1), EST223333, heme oxygenase (HO), VGF, UI-R-CO-ja-a-07-0-Ul.s1 and macrophage migration inhibitory factor (MIF) were upregulated in MCA group comparing to Control group. Expression of Putative potassium channel subunit protein (ACK4), P2X2A-5, P2X2A-4, Interferon-gamma inducing factor isoform alpha precursor (IL-18α), EST199031, P2XR, P2X2 purinoceptor isoform e (P2X2R-e), Precursor interleukin 18 (IL-18) were downregulated, whereas MOO, EST223333, GLUT-1, MIF, Neuronatin alpha, UI-R-C0-ja-a-07-0-Ul.s1, A2. adenosine receptor, COMT, neuron-specific enolase (NSE), HO, VGF, A rat novel protein which is expressed with nerve injury (E12625) were upregulated in MCR group comparing to Stressed group. The results suggest that decreased viability and AOS production of PC12 cell by H₂O₂ may be, at lease, mediated by impaired glucose transporter expression. It is implicated that the MCR treatment protect PC12 cell from oxidative stress via following mechanisms; improving glucose transport into the cell, enhancing expression of anti-oxidative genes and protecting from dopamine cytotoxicity by increment of COMT and MIF expression. The list of differentially expressed genes may implicate further insight on the action and mechanism behind the anti-oxidative effects of herbal extract Moutan Cortex Radicis.

• Clinical and Experimental Reproductive Medicine
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• v.28 no.1
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• pp.33-40
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• 2001

Objective: This study was to establish the human embryonic stem (ES) cells derived from frozen-thawed blastocyst stage embryo that were destined to be discarded after five years in routine human IVF-ET program. Methods: Frozen-thawed and survived human blastocysts were treated by immunosurgery, and recovered ICM cells were cultured onto STO feeder cell layer and ICM colony was subcultured by mechanical dissociation into clumps. To identify ES cell, alkaline phosphatase staining and expression of Oct4 in replated ICM colonies were examined. Also, to examine the possibility of ES cell differentiation, retinoic acid (RA), basic fibroblast growth factor (b-FGF), nerve growth factor (NGF) were added in culture medium. In addition, to classify the specific cell type, differentiated cells were stained by indirect immunocytochemistry. Results: One ICM colony recovered from frozen-thawed six blastocysts was subcultured, continuously replated during 40 passage culture duration without differentiation. Subcultured colonies were strong positively stained by alkaline phophatase. When the expression of Oct4 in cultured ES colony was examined, Oct4b type is more clearly indicated than Oct4a one although there was not detected in embryoid body or differentiated cells. In differentiated cardiomyocytes from ES colony, cells were beaten regularly (60 times/min). In differentiated neural cells from ES colony, neurofilament (NF) 200 kDa protein, microtubule associated protein (MAP) 2 and ${\beta}$-tubulin of specific marker in neurons, glial fibrillary acidic protein (GFAP) of specific marker in astrocytes and galactocelebrocide (GalC) of specific marker in oligodendrocytes were confirmed by indirect immunocytochemistry. Also, muscle cells were detected by indirect immunocytochemistry. In addition, ES colonies can be successfully cryopreserved. Conclusion: This study suggested that establishment of human ES cells can be successfully derived from frozen-thawed blastocysts that were destined to be discarded, and obtained specific cell types (cardiomyocytes, neurons and muscle cells) through the in vitro differentiation procedures of ES cells.

• Journal of Life Science
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• v.21 no.7
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• pp.985-991
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• 2011

The purpose of this study was to investigate the biological effect of obesity-induced oxidative damage on neurogenesis and early protein expression. Obesity was induced I thirty 4-week old male Sprague-Dawley rats through a high fat diet for 15 weeks. After one week of environmental adaptation, the rats were divided into 2 groups: high fat diet sedentary group (HDS, n=15) and high fat diet training group (HDT, n=15). Exercise training was performed 5 times a week for 8 weeks, with mild-intensity treadmill running for weeks 1-4 and moderate-intensity treadmill running for weeks 5-8. After the 8 week training period, we analyzed lipid profiles, serum 8-hydroxyguanosine (8-OHdG), liver tissue malondialdehyde (MDA) related to oxidative damage factors, nerve growth factor (NGF), brain derived neurotrophic factor (BDNF), c-fos, c-jun, and extracellular signal regulated kinase (Erk) in the hippocampus. The results of this study are as follows. There were differences between HDS and HDT in triglyceride (TG) and total cholesterol (TC) (p<0.05). In high density lipoprotein (HDL-c), the HDT was higher than HDS after treadmill training (p<0.05). In 8-OHdG, the HDT was lower than HDS after treadmill training (p<0.05). Genetic expressions of c-jun, BDNF and MDA in the HDT were higher than in the HDS after treadmill training in hippocampus (p<0.05). Therefore, we conclude that 8 weeks of treadmill training can improve imbalanced lipid profiles, reduce oxidative damage, and activate neurogenesis in obese rats.

• Journal of Physiology & Pathology in Korean Medicine
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• v.18 no.4
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• pp.1041-1054
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• 2004

Electroacupuncture (EA) has been reported to increase pain threshold, and to enhance the NK cell activity by up-regulation of IFN-γ and endogenous β-endolphin. For the purpose of understanding the molecular mechanism of EA stimulation, we analyzed the gene expression profile of rat hypothalamus, treated on Zusanli (ST36) with EA, in comparison with control group by oligonucleotide chip microarray (Affymetrix GeneChip Rat Neurobiology U34 Array) and real-time RT-PCR. Sprague-Dawley (S-D) male rats were stimulated at the Zusanli (ST36) acupoint in restriction holder. Simultaneously the control group was given only holder stress without EA stimulation. In order to prove the appropriateness of EA treatment, we measured spleen NK cell activity with standard 51Cr release assay. NK cell activity of EA group was significantly increased comparing to control group. The microarray and PCR results show that EA treatment up-regulates expression of genes associated with 1) nerve growth such as NGF induced factor A and VGF, 2) signal transduction such as 5HT3 receptor subunit, AMPA receptor binding protein and Na-dependent neurotransmitter transporter, and 3) anti-oxidation such as superoxide dismutase and glutathione S-transferase. In addition, the activity of the anti-oxidative enzyme, SOD of hypothalamus, liver and RBC was enhanced compared to that of control. The list of differentially expressed genes may implicate further insight on the mechanism of acupuncture effects.