• Applied Microscopy
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• v.16 no.2
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• pp.1-13
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• 1986

The purpose of this study was to investigate the morphology and intercellular junctions of the odontoblast of dental pulp in the rat incisor by means of the freeze fracture electron microscopy. Twenty male Sprague-Dawley rats weighing $150{\sim}200g$ were used. After being anesthetized by an intraperitoneal injection of 0.5 ml sodium pentobarbital per kg in body weight(60 mg/ml) the animals were perfused with 2.5% glutaraldehyde-2% paraformaldehyde fixative in 0.1 M cacodylate buffer, pH 7.2 through the ascending aorta for one hour. The incisors were carefully extracted from the jaws and demineralized by suspending them in 0.1 M EDTA in 3% glutaraldehyde (pH 7.2) for two weeks. After demineralization, the specimens were obtained from the portion divided into five equal parts. For freeze-fracture replication, demineralized tissues were infiltrated for several hours with 10%, 25% glycerol in 0.1M cacodylate buffer as a cryoprotectant and then frozen in liquid Freon 22 and stored in liquid nitrogen. Fracturing and replication were done in Balzers BAF 400D high-vacuum freeze-fracture apparatus at $-120^{\circ}C$ under routine $5X10^{-7}$ Torr vacuum. The tissue was immediately replicated with platinum unidirectionally at $45^{\circ}$ angle and reinforced with carbon at $90^{\circ}$ angle unidirectionally or by using a rotary stage. The replication process was monitored by a quartz-crystal device. The replicas were immersed in 100% methanol overnight. The tissue was then digested from the replica by clorox (laundry bleach), placed into 5% EDTA, and washed repeatedly with distilled water. The replicas were picked up on 0.3% formvar-coated 75 mesh grids and examined in the JEOL 100B electron microscope. The results were as follows; 1. Both in thin sections and freeze-fracture replicas, three types of intercellular junctions were recognizable in the plasma membrane of odontoblast: gap junction, tight junction and desmosome-like junction. 2. The nuclear pores were evenly distributed over the nuclear envelope. The pore complex formed a ring about 70 nm in diameter. 3. Gap junctions were found between odontoblasts as well as odontoblasts and neighbouring pulp cells (fibroblast, subodontoblastic cell process, nerve-like fibre). Gap junctions, which were round, ellipsoid and pear-shaped and 600 nm in diameter, were observed in the odontoblast. 4. Numerous round and ellipsoid gap junctions could be frequently seen on the plasma membranes in cell body and apical part of the odontoblasts. On the P face, the junctions were recognized as a cluster of closely packed particles, measuring about 9 nm in diameter, and on the E face, the junctions were recognized as a shallow grooves.

• Applied Microscopy
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• v.39 no.4
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• pp.325-332
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• 2009

It has been well known that grapefruit seed extract (GSE) have effects on the deodorization and a wide range disinfectant. But anybody do not research about the decomposition-inhibition effect of GSE. The present study was performed to investigate the decomposition-inhibition effect of GSE by measuring the microscope and electroscope observation with passage of time. A total of 30 healthy Sprague-Dawley (SD) rats weighting from 230 gm to 250 gm were used as experimental animals. One group that do not treated by GSE named control group and the other group that treated by 55% GSE named Experimental group. Under ether anesthesia, right kidney was obtained. Put each sample in $37^{\circ}C$ and humidity $80{\pm}5%$ incubator, we obtained each sample after 0 day, 1 day, 2 days, 3 days, 4 days and 5 days. 4 ${\mu}m$ of paraffin sections were obtained, stained H-E stain and observed by use of a microscope. Morphological change in tissue was similar to control 1 day group and experimental 3 days group. Therefore, decomposition-inhibition effect of GSE continued about 2 days and it protected necrosis. According to above results, the author suggest that 55% GSE is an effective decomposition inhibitor until 2 days on $37^{\circ}C$ and humidity $80{\pm}5%$ conditions.

• Journal of Physiology & Pathology in Korean Medicine
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• v.16 no.5
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• pp.1025-1034
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• 2002

The herbal extract (YMT_02) is a modified herbal extracts from Yukmijihwang-tang (YMJ) to promote memory-enhancing. The YMJ extracts has been widely used as an anti-aging herbal medicine for hundred years in Asian countries. The purpose of this study is to; 1) quantitatively evaluate the memory-enhancing effect of YMT_02 by behavior task, 2) identify candidate genes responsible for enhancing memory by cDNA microarray and 3) assess the anti-oxidant effect of YMT_02 on PC12 cell. Memory retention abilities are addressed by passive avoidance task with Sprague-Dawley (SD) male rat. Before the training session, the rats are subdivided into four groups and administrated with YMT_02, Ginkgo biloba, Soya lecithin and normal saline for 10 days. The retention test was performed. 24 hours after the training session. The retention time of the YMT_02 group was significantly (p<0.05) delayed (～100%), whereas Ginkgo biloba and Soya lecithin treatment delayed 20% and 10% respectively. The hippocampi of YMT_02 and control group were dissected and mANA was further purified. After synthesizing cDNA using oligo-dT primer, the cDNA were applied to Incyte rat GEMTM 2 cDNA microarray. The microarray results show that prealbumin(transthyretin), phosphotidylethanolamine N-methyltransferase, and PEP-19 are expressed abundantly in the YMT_02 treated group. Especially, PEP-19 is a neuron-specific protein, which inhibits apoptotic processes in neuronal cell. On the other hand, transcripts of RAB15, glutamate receptor subunit 2 and CDK108 are abundant in control group. Besides, neuronal genes involved in neuronal death or neurodegeneration such as neuronal-pentraxin and spectrin are abundantly expressed in control group. Additionally, the YMT_02 shows an anti oxidative effect in the PC12 cell. The list of differentially expressed genes may implicate further insight on the action and mechanism behind the memory-enhancing effect of herbal extracts YMT_02, for example, anti-apoptotic, anti-oxidative, and neuroprotective effects.

• Science of Emotion and Sensibility
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• v.11 no.4
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• pp.565-574
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• 2008

Functions of human brain including sensibility and emotion may be affected by drugs mediated by the blood-brain barrier (BBB). The present study was performed to evaluate whether injection route, volume and concentration of mannitol could alter the degree of disruption of the BBB. Under urethane anesthesia, female Sprague-Dawley rats were infused with 20% mannitol into the right internal carotid artery (ICA). In the other group, intravenous injection of mannitol through the femoral vein was performed. Evans blue(EB) dye was used as a marker of BBB disruption. When mannitol was injected via the ICA, the content of EB dye in the ipsilateral hemisphere was markedly increased. However, the content of EB in the brain was not increased when mannitol was injected via the femoral vein, even though the volume or concentration of mannitol was increased. These results suggest that the BBB was disrupted only through ICA injection route and this may provide a useful strategy for transient opening of the BBB to control the functions of human brain.

• Journal of dental hygiene science
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• v.17 no.1
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• pp.65-72
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• 2017

Temporomandibular joint (TMJ) pain is characterized by persistent jaw pain associated with dysfunction and tenderness of the temporomandibular muscles and joints. The aim of this study was to investigate whether treatment with red or black ginseng extract helps in the modulation of inflammatory TMJ pain. Male Sprague-Dawley rats weighing 220~260 g were used. The experimental group was subdivided into 4 groups based on the treatment method (n=6, each group): formalin (5%, $30{\mu}l$), formalin after distilled water (vehicle), formalin after red or black ginseng extract (per oral, single or repeated, respectively). To induce TMJ pain, $30{\mu}l$ of formalin was injected into the articular cavity under ether inhalation anesthesia. The number of noxious behavioral responses of scratching the facial region proximal to the injection site was recorded for 9 successive 5-min intervals following formalin injection. Repeated treatment with red or black ginseng extract reduced the nociceptive responses in the second phase (11~45 min). Nuclear factor erythroid 2-related factor 2 (Nrf2) is an oxidative stress-mediated transcription factor. Both ginsengs significantly down-regulated the increased Nrf2 level compared to the vehicle group. In the test for liver and kidney functions, repeated treatment with red or black ginseng was not different compared to the vehicle group. These results indicate that red and black ginseng extract might be promising analgesic agents in the treatment of inflammatory TMJ pain.

• Journal of the Korean Society of Food Science and Nutrition
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• v.31 no.1
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• pp.87-91
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• 2002

This study was designed to investigate the effects of Hijikia fusiforme (Harvey) Okamura ethanol extract on the ethanol-induced hepatotoxicity of rat administered orally experimental diets for 6 weeks. Sprague-Dawley rats weighing about 100 g were divided into 4 groups; normal group (NOR), ethanol (35% ethanol 10 mL/kg b.w/day) treated group (CON), ethanol and Hijikia fusiforme ethanol extract 200 mg/kg (HE1) and 400 mg/kg (HE2) concomitantly treated group, respectively. Each group was examined for the growth rate, feed efficiency ratio (FER), activities of antioxidative enzymes and contents of TBARS and glutathione. Hijikia fusiforme ethanol extract showed increasing effects of the growth rate by 43%, and FER was gradually increased by Hijikia fusiforme ethanol extract treatment, compard with ethanol treatment. Ethanol elevated the activities of superoxide dismutase, catalase and glutathione peroxidase of rat liver markedly as compared to normal group, but those activities were significantly decreased in Hijikia fusiforme ethanol extract treatment by 56%, 38% and 25%, respectively. Xanthine oxidase activity elevated by ethanol was not affected by Hijikia fusiforme ethanol extract. The content of TBARS increased by ethanol treatment was signigicantly decreased in HE2, and the glutathione content depleted by ethanol treatment was increased by Hijikia fusiforme ethanol extract administration adjacent to normal level. These results suggest that Hijikia fusiforme ethanol extract is believed to be a possible protective effect for the ethanol-induced hepatotoxicity of rat liver.

• Korean Journal of Acupuncture
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• v.24 no.4
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• pp.201-219
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• 2007

Objectives : We studied Effects of Hagocho(prunella vulgaris L.), Gamgook(chrysanthemum indicum L.) and Galgeun(pueraria Radix) aqua-acupuncture on the hyperlipidemic rat. methods : We investigated lowering lipid effect, oxidative capacity, concentration of TNF-${\alpha}$, IL-6, Leptin and histological consideration in hyperlipidemic rat. Forty male Sprague-Dawley rats weighing about 400g were divided into 5 groups of control, Ⅰ: Hagocho (prunella vulgaris L.)+Gamgook (chrysanthemum indicum L.) and Gokji aqua-acupuncture, Ⅱ: Hagocho(prunella vulgaris L.)+Gamgook (chrysanthemum indicum L.) and Joksamri aqua-acupuncture, Ⅲ: Hagocho(prunella vulgaris L.)+Galgeun(pueraria Radix)and Gokji aqua-acupuncture and Ⅳ: Hagocho (prunella vulgaris L.)+Galgeun(pueraria Radix) and Joksamri aqua-acupuncture. Results : Contents of plasma ${\beta}$-lipoprotein, contents of IL-6, TNF-${\alpha}$ and leptin, Plasma triglyceride and glucose, plasma total cholesterol and LDL-cholesterol, liver total cholesterol, liver triglyceride, plasma and liver TBARS, free fatty acids showed a tendency to decrease in the aqua-acupuncture groups compared to those of control group. The activities of GOT and GPT showed no significantly different in all treatment groups. Values of super oxide dismutase, glutathione peroxidase and catalase activity showed a tendency to increase in the aqua-acupuncture groups. Histological consideration of heart, kidney and liver in the aqua-acupuncture groups showed slight vasodilation and fat accumulation compared to those of normal rat. Conclusions : These results indicated that prunella vulgaris L., chrysanthemum indicum L. and pueraria Radix aqua-acupuncture at gokji(LI11) and Joksamri(ST36) suppressed adipose tissue mass and lipid peroxidation and increased antioxidant system in hyperlipidemic rat.

• Toxicological Research
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• v.29 no.3
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• pp.173-179
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• 2013

In-utero exposure to valproic acid (VPA) has been known as a potent inducer of autism spectrum disorder (ASD), not only in humans, but also in animals. In addition to the defects in communication and social interaction as well as repetitive behaviors, ASD patients usually suffer from gastrointestinal (GI) problems. However, the exact mechanism underlying these disorders is not known. In this study, we examined the gross GI tract structure and GI motility in a VPA animal model of ASD. On embryonic day 12 (E12), 4 pregnant Sprague-Dawley (SD) rats were subcutaneously injected with VPA (400 mg/kg) in the treatment group, and with phosphate buffered saline (PBS) in the control group; the resulting male offspring were analyzed at 4 weeks of age. VPA exposure decreased the thickness of tunica mucosa and tunica muscularis in the stomach and ileum. Other regions such as duodenum, jejunum, and colon did not show a significant difference. In high-resolution microscopic observation, atrophy of the parietal and chief cells in the stomach and absorptive cells in the ileum was observed. In addition, decreased staining of the epithelial cells was observed in the hematoxylin and eosin (H&E)-stained ileum section. Furthermore, decreased motility in GI tract was also observed in rat offspring prenatally exposed to VPA. However, the mechanism underlying GI tract defects in VPA animal model as well as the association between abnormal GI structure and function with ASD is yet to be clearly understood. Nevertheless, the results from the present study suggest that this VPA ASD model undergoes abnormal changes in the GI structure and function, which in turn could provide beneficial clues pertaining to the pathophysiological relevance of GI complications and ASD phenotypes.

• Toxicological Research
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• v.34 no.4
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• pp.333-341
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• 2018

Ferulate is a phenolic compound abundant in wheat germ and bran and has been investigated for its beneficial activities. The aim of the present study is to evaluate the efficacy of ferulate against the oxidative stress-induced imbalance of protein tyrosine kinases (PTKs), protein tyrosine phosphatases (PTPs), and serine/threonine protein phosphatase 2A (PP2A), in connection with our previous finding that oxidative stress-induced imbalance of PTKs and PTPs is linked with proinflammatory nuclear factor-kappa B $(NF-{\kappa}B)$ activation. To test the effects of ferulate on this process, we utilized two oxidative stress-induced inflammatory models. First, YPEN-1 cells were pretreated with ferulate for 1 hr prior to the administration of 2,2'-Azobis(2-methylpropionamidine) dihydrochloride (AAPH). Second, 20-month-old Sprague-Dawley rats were fed ferulate for 10 days. After ferulate treatment, the activities of PTKs, PTPs, and PP2A were measured because these proteins either directly or indirectly promote $NF-{\kappa}B$ activation. Our results revealed that in YPEN-1 cells, ferulate effectively suppressed AAPH-induced increases in reactive oxygen species (ROS) and $NF-{\kappa}B$ activity, as well as AAPH-induced PTK activation. Furthermore, ferulate also inhibited AAPH-induced PTP and PP2A inactivation. In the aged kidney model, ferulate suppressed aging-induced activation of PTKs and ameliorated aging-induced inactivation of PTPs and PP2A. Thus, herein we demonstrated that ferulate could modulate PTK/PTP balance against oxidative stress-induced inactivation of PTPs and PP2A, which is closely linked with $NF-{\kappa}B$ activation. Based on these results, the ability of ferulate to modulate oxidative stress-related inflammatory processes is established, which suggests that this compound could act as a novel therapeutic agent.

• 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.