• Toxicological Research
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• v.25 no.2
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• pp.71-78
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• 2009

We investigated the pathogenesis of liver tissue damage during the lipid peroxidation and fibrogenesis with the observation of correlations between the parameters of collagen synthesis (and deposition) and lipid peroxidation in liver fibrosis (cirrhosis) rats. Rats were randomly divided into two groups, normal and $CCl_4$-thiopental sod. intoxicated group. And the one group was treated intragastrically with the mixture of $CCl_4$-thiopental sod. 3 times per week for 3 weeks. The liver tissue and sera were used for the measurement of hydroxyproline (HYP), malonedialdehyde (MDA) and superoxide dismutase (SOD). Biochemical parameters such as aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), total-bilirubin and blood urea nitrogen (BUN) were measured. Additionally, the expression of collagen ${\alpha}1$(III) and $\beta$-actin mRNA was observed by RTPCR. The histological change in liver tissue was also observed by Masson's trichrome and H&E staining. Correlation analysis was carried by Spearman's rho method. All biochemical parameters except total-bilirubin were significantly higher in the $CCl_4$-thiopental sod. treated group than that of the normal group (p < 0.01). In the $CCl_4$-thiopental sod. treated group, Hyp as a parameter of collagen synthesis (deposition) and MDA as a metabolite of lipid peroxidation, were significantly elevated by 1.98 and 2.11 times higher than that of the normal group (p < 0.001) respectively. The activity of SOD in the $CCl_4$-thiopental sod. treated group is decreased significantly by 44.8% (p < 0.001). And collagen ${\alpha}1$(III) mRNA was more expressed in the $CCl_4$-thiopental sod. treated group than that of the normal group. However, the expression of $\beta$-actin mRNA is showed similar in both of groups. A good correlation was observed between the content of hyp and MDA concentration (r = 0.70, n = 40) in the two groups. And the correlation between the levels of hyp and SOD (r = -0.71, n = 25) is also reliable. However, no correlation were observed between MDA concentration and SOD (r = -0.40, n = 25) in the two groups. Elevated levels of MDA in $CCl_4$-thiopental sod. treated rats indicated enhancement of lipid peroxidation, which is accompanied by a decrease in SOD activity. Moreover, we could confirm that the parameters of collagen synthesis (and deposition) is in good correlation with the metabolite of lipid peroxidation (MDA) and the lipid peroxidation antagonizing enzyme (SOD). Hence, we propose that (1) lipid peroxidation and collagen synthesis (and deposition) could be enhanced by intragastrically application of $CCl_4$-thiopental sod. during a short terms. And (2) the intoxication of $CCl_4$-thiopental sod. could be used for monitoring of lipid peroxidation and collagen synthesis (and deposition) for test of antioxidant and antifibrotic agent.

• The Korean Journal of Physiology and Pharmacology
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• v.22 no.6
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• pp.607-616
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• 2018

The effect of melatonin on juveniles with cardio fibrosis is poorly understood. We investigated whether HDACs participate in the anti-fibrotic processes regulated by melatonin during hypertrophic remodeling. Abdominal aortic constriction (AAC) was employed in juvenile rats resulting in pressure overload-induced ventricular hypertrophy and melatonin was subsequently decreased via continuous light exposure for 5 weeks after surgery. AAC rats displayed an increased cross-sectional area of myocardial fibers and significantly elevated collagen deposition compared to sham-operated rats, as measured by HE and Masson Trichrome staining. Continuous light exposure following surgery exacerbated the increase in the cross-sectional area of myocardial fibers. The expression of HDAC1, HDAC2, HDAC3, HDAC4 and HDAC6 genes were all significantly enhanced in AAC rats with light exposure relative to the other rats. Moreover, the protein level of $TNF-{\alpha}$ was also upregulated in the AAC light exposure groups when compared with the sham. However, Smad4 protein expression was unchanged in the juveniles' hearts. In contrast, beginning 5 weeks after the operation, the AAC rats were treated with melatonin (10 mg/kg, intraperitoneal injection every evening) or vehicle 4 weeks, and sham rats were given vehicle. The changes in the histological measures of cardio fibrosis and the gene expressions of HDAC1, HDAC2, HDAC3, HDAC4 and HDAC6 were attenuated by melatonin administration. The results reveal that melatonin plays a role in the development of cardio fibrosis and the expression of HDAC1, HDAC2, HDAC3, HDAC4 and HDAC6 in cardiomyocytes.

• Nutrition Research and Practice
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• v.14 no.4
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• pp.334-351
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• 2020

BACKGROUND/OBJECTIVES: This study was designed to investigate the improvement effect of white ginseng extract (GS-KG9) on D-galactosamine (Ga1N)-induced oxidative stress and liver injury. SUBJECTS/METHODS: Sixty Sprague-Dawley rats were divided into 6 groups. Rats were orally administrated with GS-KG9 (300, 500, or 700 mg/kg) or silymarin (25 mg/kg) for 2 weeks. The rats of the GS-KG9- and silymarin-treated groups and a control group were then intraperitoneally injected Ga1N at a concentration of 650 mg/kg for 4 days. To investigate the protective effect of GS-KG9 against GalN-induced liver injury, blood liver function indicators, anti-oxidative stress indicators, and histopathological features were analyzed. RESULTS: Serum biochemical analysis indicated that GS-KG9 ameliorated the elevation of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), and lactate dehydrogenase (LDH) in GalN-treated rats. The hepatoprotective effects of GS-KG9 involved enhancing components of the hepatic antioxidant defense system, including glutathione, glutathione peroxidase (GPx), superoxide dismutase (SOD), and catalase (CAT). In addition, GS-KG9 treatment inhibited reactive oxygen species (ROS) production induced by GalN treatment in hepatocytes and significantly increased the expression levels of nuclear factor erythroid-2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) proteins, which are antioxidant proteins. In particular, by histological analyses bases on hematoxylin and eosin, Masson's trichrome, α-smooth muscle actin, and transforming growth factor-β1 staining, we determined that the administration of 500 mg/kg GS-KG9 inhibited hepatic inflammation and fibrosis due to the excessive accumulation of collagen. CONCLUSIONS: These findings demonstrate that GS-KG9 improves GalN-induced liver inflammation, necrosis, and fibrosis by attenuating oxidative stress. Therefore, GS-KG9 may be considered a useful candidate in the development of a natural preventive agent against liver injury.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.30 no.5
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• pp.407-416
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• 2008

Purpose: The aim of the present study is to evaluate the effect of repetitive distraction and compression on new bone formation during distraction period. Materials and methods: Sixteen healthy rabbits, weighing about 2.5kg, were used in this experiment. A unilateral mandibular osteotomy was performed in the left mandible and the distractor(Track 1 $plus^{(R)}$, Gebruder Martin $GmbH^{(R)}$, Germany) was fixed with four screws (Cross driver screw $TI^{(R)}$, Gebruder Martin $GmbH^{(R)}$, Germany). After 4 days, the mandibles were distracted at a rate of 0.6mm/day for 10 days to obtain the amount of 6mm distraction in the control group(n=4). In the experimental group A(n=6), they were distracted at a rate of 1.2mm/day for 5 days and then compression of 0.6mm length and distraction of 0.6mm per 12 hours were carried out as counter direction for 5 days, relatively. In the experimental group B(n=6), distraction of 1.2mm length and compression of 0.6mm length per 12 hours were repeated for 10 days to obtain the amount of 6mm distraction finally. The experimental animals were sacrificed at 2 and 4 weeks after surgery and block specimens were obtained. With histologic and histomorphometric analysis, we observed the histologic changes of the cells and bone formation after H-E and Masson- Trichrome staining and then, measured Bone Deposition Rate with TOMORO $ScopeEye^{TM}$ ver. 3.5(Olympus, Japan), Results: Histologically, new bone formation was examined in all experimental groups and the control. But, the ability of bone formation of the experimental group A was somewhat better than any other groups. On the histomorphometric analysis, Bone Deposition Rate was higher in the experimental group A$(50.67{\pm}4.36%)$ than in the control group$(45.94{\pm}3.97%)$ and in the experimental group B$(42.68{\pm}5.70%)$. These data showed significant differences statistically(p<0.05). Conclusion: These results show that the distraction osteogenesis using repetitive compression and distraction force in the early consolidation period may be effective for new bone formation.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.33 no.6
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• pp.459-466
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• 2011

Purpose: This study evaluated the capability of bone formation with silk fibroin/nano-hydroxyapatite/corn starch composite scaffold as a bone defect replacement matrix when grafted in a calvarial bone defect of rabbits $in$ $vivo$. Methods: Ten New Zealand white rabbits were used for this study and bilateral round-shaped defects were formed in the parietal bone (diameter: 8.0 mm). The silk fibroin 10% nano-hydroxyapatite/30% corn starch/60% composite scaffold was grafted into the right parietal bone (experimental group). The left side (control group) was grafted with a nano-hydroxyapatite (30%)/corn starch (70%) scaffold. The animals were sacrificed at 4 weeks and 8 weeks. A micro-computerized tomography (${\mu}CT$) of each specimen was taken. Subsequently, the specimens were decalcified and stained with Masson's trichrome for histological and histomorphometric analysis. Results: The average ${\mu}CT$ and histomorphometric measures of bone formation were higher in the control group than in the experimental group at 4 weeks and 8 weeks after surgery though not statistically significant ($P$ >0.05). Conclusion: The rabbit calvarial defect was not successfully repaired by silk fibroin/nano-hydroxyapatite/corn starch composite scaffold and may have been due to an inflammatory reaction caused by silk powder. In the future, the development of composite bone graft material based on various components should be performed with caution.

• The korean journal of orthodontics
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• v.28 no.3 s.68
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• pp.453-459
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• 1998

The dental implants for edentulous Patients have been used for more than 20 years. After the introduction of osseointegration by $Br{\aa}nemark$, the commercially pure titanium implants were accepted by most practitioners. Recently dental implants are used for orthodontic anchorages as well as prosthetic abutment. Many researchers have reported implants as a good orthodontic anchorage through basic research and clinical evaluation. But previous researches were done after the healing time for osseointegration of inserted implants. If dental implants are to be used for prosthetic abutment the healing time for osseointegration is necessary, but orthodontic forces to implants are different from bite force regarding its amount of force, duration and direction. The authors evaluated the effect of orthopedic force to implants on bone tissue before osseointegration. 48 implants were placed at 12 rabbits. 2 implants into left side and 2 implants into right side were inserted along the long axis of femur respectively 2 weeks (2 weeks group), 4 weeks (4 weeks group) and 6 weeks (6 weeks group) after implants placement, 300g force had been applied to the implants at left side femur by Ni-Ti close coil spring for 4 weeks (experimental group) and no force applied to implants at right side femur (control group). After the force application for 4weeks, rabbits were sacrificed and microscopic evaluation was done by hematoxylin-eosin stain and Masson trichrome stain. The result3 were followed. 1. All implants in experimental group remained rigid after the force application for 4 weeks. 2. More fibrous tissue between bone and implants were noticed at 2 weeks experimental group than 2 weeks control group 3. More bone remodeling was noticed at 4weeks group than 2 weeks group and it was difficult to find out fibrous tissue between bone and implants at both experimental and control group of 4 weeks group. 4. It was hard to distinguish experimental group from control group at 6 weeks group. Therefore if initial stability can be obtained on implant insertion, it can be possible to use implants as a orthodontic anchorage before the healing time for osseointegration.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.27 no.1
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• pp.15-24
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• 2001

The purpose of this study is to evaluate the tissue response in applying of various bone substitutes included toothash-plaster mixture, resorbable hydroxylapatite (HA) and demineralized freeze-dried bone and to show the clinical usefulness of toothash-plaster mixture for the repair of craniomaxillofacial bone defect. For this experiment, 100 Sprague-Dawley rats weighing 200gm or more were used. There were four experimental groups: group I, toothash-plaster mixture; group II, demineralized freeze-dried bone; group III, resorbable HA; and group IV, control group. A full thickness, round bone defect measuring 10mm in diameter was created in the midcranium, and the substitutes cited above were embedded in the experimental rats based on their group assignment. Blood clot was filled in the rats assigned to the control group. Experimental rats were sacrificed on the 1st, 3rd, 5th, 8th, 12th and 24th week after implantation and stained with the hematoxylineosin, Masson's Trichrome, using Van Gieson's stain method, and were examined under light microscope. The results were as follows: 1. In all the groups, prominent inflammatory reaction and the infiltration of multinucleated giant cells were noted during the early stage. Gradual healing decreased this reaction. 2. Among the rats in the experimental group II, which were given demineralized freeze-dried bone implants, active formation of new bone traveculae manifested. Chondroid tissues appeared, and it was suggested that the defect was filled with newly formed bone by virtue of osteoinductive activity. On the 12th week after the experiments, most of the defect was filled with newly formed bone trabeculae. 3. In experimental groups I and III, it was noted that HA manifested a healing process similar to that characterized by the toothash-plaster mixture, but inflammatory reaction was more prominent in experimental group I. Active osteoblasts were observed along the periphery of osteoid tissues, while newly formed bone trabeculae appeared adjacent to the implanted materials three weeks later. Formation increased to the extent that newly formed bone trabeculae fused directly with the host bone. Increase in new bone ingrowth into the filling materials was revealed by both experimental groups. 4. In the control group, new bone formation adjacent to the host bone was observed, but most of the defect was filled with mature connective tissue 24 weeks after the experiments.

• Journal of Periodontal and Implant Science
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• v.26 no.4
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• pp.967-987
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• 1996

The present study was to evaluate the healing patterns of guided tissue regeneration( GTR) using resorbable $Vicryl^{(R)}$(polyglactin 910) mesh and nonresorbable expanded polytetrafluoroethylene(ePTFE) membrane with or without bone grafting using autogeneous bone and demineralized freeze-dried bone allograft(DFDBA) in the grade II furcation defects. Mucoperiosteal flaps were reflected buccally in the mandibular 2nd, 3rd and 4th premolar areas and furcation defects were created surgically by removing $5{\times}6mm$ alveolar bone in 4 dogs. Root surfaces were thoroughly debrided of periodontal ligament and cementum, and notches were placed on root surface at the most apical bone level. In the right and left mandibular quadrant, each tooth was received $Vicryl^{(R)}$ mesh(ACE Surgical Supply Co., USA) only, $Vicryl^{(R)}$ mesh with DFDBA, $Vicryl^{(R)}$ mesh with autogeneous bone grafts, ePTFE membrane($Core-tex^{(R)}$ membrane, W.L. Gore & Associates Inc., USA) only, ePTFE membrane with DFDBA or ePTFE membrane with autogeneous bone grafts. For the fluorescent microscopic examination, fluorescent agents were injected at 2, 4 and 8 weeks after surgery. Four weeks after surgery, 2 dogs were sacrificed and ePTFE membranes were removed from remaining 2 dogs, which were sacrificed at 12 weeks after surgery. Undecalcified tissues were embedded in methylmethacrylate and $10{\mu}m$ thick sections were cut in a buccolingual direction. These sections were stained with hematoxylin-eosin stain and Masson's trichrome stain, and evaluated by descriptive histology and linear measurements. The results were as follows : 1) $Vicryl^{(R)}$ mesh group showed less connective tissue attachment than ePTFE membrane group. 2) The combination of GTR using $Vicryl^{(R)}$ mesh and osseous grafts resulted in new attachment and new bone formation more than GTR using $Vicryl^{(R)}$ mesh only. 3) GTR using ePTFE membrane, with or without osseous grafts, enhanced periodontal regeneration. 4) Root resorption and dentoalveolar ankylosis were observed in the areas treated with the combination of GTR and DFDBA. It was suggested that the effect of adjunctive bone grafting in GTR procedure depends on the materials and the physical properties of barrier membranes. $Vicryl^{(R)}$ mesh performed a barrier function and the use of adjunctive bone grafting may enhance the periodontal regeneration.

• The Journal of Korean Medicine
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• v.34 no.2
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• pp.51-58
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• 2013

Objectives: The aim of this study was to examine the antithrombotic effects of the four herbal ingredients (Mume Fructus, MF; Santali Albi Lignum, SAL; Amomi Tsao-Ko Fructus, ATF; and Amomi Fructus, AF) of modified Jeho-tang (MJHT) in a ferric chloride ($FeCl_3$)-induced carotid arterial thrombosis model. Methods: Thirty minutes prior to a 35% $FeCl_3$ application, Sprague-Dawley (SD) rats were injected with saline, MF, SAL, ATF or AF (100 mg/kg, intraperitoneal injection), respectively. The effect of the MJHT ingredients was examined for time to occlusion (TTO) and thrombus weight (TW) in a $FeCl_3$-induced thrombosis model. Histological analysis was performed to examine the effect of the MJHT ingredients on collagen fiber damage using hematoxylin & eosin and Masson's trichrome staining. Results: Compared with vehicle treatment, MF, SAL and ATF treatment delayed TTO (vehicle, $8.11{\pm}0.60$ min; MF, $16.67{\pm}1.03$ min; SAL, $17.50{\pm}1.52$ min and ATF, $13.33{\pm}1.21$ min; P < 0.001) and inhibited thrombus formation (vehicle, $0.79{\pm}0.03$ mg/mm; MF, $0.61{\pm}0.07$ mg/mm; SAL, $0.57{\pm}0.03$ mg/mm and ATF, $0.72{\pm}0.02$ mg/mm; P < 0.001). In addition, each herbal ingredient of MJHT except for AF prevented the collagen fiber damage induced by a 35% $FeCl_3$ application. These results indicate that the MJHT ingredients MF ${\geq}$ SAL ${\geq}$ ATF ${\geq}$ AF possess antithrombotic activity in a $FeCl_3$-induced carotid arterial thrombosis. Conclusions: Altogether, these results are the first evidence that the MJHT ingredients MF, SAL and ATF have the ability to prevent vascular damage and thrombus formation in $FeCl_3$-induced carotid arterial thrombosis.

• Journal of Physiology & Pathology in Korean Medicine
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• v.23 no.5
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• pp.1003-1011
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• 2009

This study was carried out to know the effects of Kwanjulbang-6(GJB-6) on the inhibition of arthritis. GJB-6 was orally administered to mouse with arthritis induced by collagen II . Cytotoxicity, hepatotoxicity, arthritis index, value of immunocyte in draining lymph node and paw joint, rheumatoid factor in serum were measured in vivo. The incidence of arthritis was significantly decreased. Total cell number of draining lymph node was significantly increased compared with control. Total cell number of paw joint was significantly decreased compared with control. The absolute number of $CD19^+$, $CD8^+$, $CD3^+/CD69^+$, $CD3^+/CD49b^+$, $CD4^+/CD44^+$, $CD3^+$, $CD4^+$, $CD4^+/CD25^+$ and $CD3^+/CD8^+$ cells in draining lymph node were significantly increased compared with control. The absolute number of $CD3^+$, $CD4^+$, $CD4^+/CD25^+$ and $CD11b^+/Gr-1^+$ cells in paw joint were significantly decreased compared with control. The absolute number of $B220^+/CD23^+$ and $MHCII^+/CD11c^+$ cell in draining lymph node were significantly decreased compared with control. The levels of IgG was decreased and The levels of IgM was significantly decreased compared with control. Anti-collagen II in serum was significantly decreased compared with control. With the hematoxylin and eosin stain, the cartilage destruction and synovial cell proliferation were decreased compared with control. With the Masson's trichrome stain, the expression of collagen fibers was decreased compared with control. Results showed that GJB-6 had immunomodulatory effects. So we expect that GJB-6 should be used as a effective drugs for not only rheumatoid arthritis but also another auto-immune disease.