• Journal of Periodontal and Implant Science
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• 제34권1호
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• pp.205-221
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• 2004

Periodontal ligament(PDL) cells have been known as playing an important roles in periodontal regeneration and gingival fibroblasts are also important to periodontal regeneration by forming connective tissue attachment. There were rare studies about the gene expression patterns of PDL cells and gingival fibroblasts, therefore in this study, we tried cDNA microarray-based gene expression monitoring to explain the functional differences of PDL cells and gingival fibroblasts in vivo and to confirm the characteristics of PDL cells. Total RNA were extracted from PDL cells and gingival fibroblasts of same person and same passages, and mRNA were isolated from the total RNA using Oligotex mRNA midi kit(Qiagen) and then fluorescent cDNA probe were prepared. And microarray hybridization were performed. The gene expression patterns of PDL cells and gingival fibroblasts were quite different. About 400 genes were expressed more highly in the PDL cells than gingival fibroblasts and about 300 genes were more highly expressed in the gingival fibroblasts than PDL cells. Compared growth factor- and growth factor receptor-related gene expression patterns of PDL cells with gingival fibroblasts, IGF-2, IGF-2 associated protein, nerve growth factor, placental bone morphogenic protein, neuron-specific growth- associated protein, FGF receptor, EGF receptor-related gene and PDGF receptor were more highly expressed in the PDL cells than gingival fibroblasts. The results of collagen gene expression patterns showed that collagen type I, type III, type VI and type VII were more highly expressed in the PDL cells than gingival fibroblasts, and in the gingival fibroblasts collagen type V, XII were more highly expressed than PDL cells. The results of osteoblast-related gene expression patterns showed that osteoblast specific cysteine-rich protein were more highly expressed in the PDL cells than gingival fibroblasts. The results of cytoskeletal proteins gene expression patterns showed that a-smooth muscle actin, actin binding protein, smooth muscle myosin heavy chain homolog and myosin light chain were more highly expressed in the PDL cells than gingival fibrobalsts, and ${\beta}-actin$, actin-capping protein(${\beta}$ subunit), actin- related protein Arp3(ARP) and myosin class I(myh-1c) were more highly expressed in the gingival fibroblasts than PDL cells. Osteoprotegerin/osteoclastogenesis inhibitory factor(OPG/OCIF) was more highly expressed in the PDL cells than gingival fibroblasts. According to the results of this study, PDL cells and gingival fibroblasts were quite different gene expression patterns though they are the fibroblast which have similar shape. Therefore PDL cells & gingival fibroblasts are heterogeneous populations which represent distinct characteristics. If more studies about genes that were differently expressed in each PDL cells & gingival fibroblasts would be performed in the future, it would be expected that the characteristics of PDL cells would be more clear.

• Journal of Chest Surgery
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• 제36권2호
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• pp.86-90
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• 2003

폐 형성에 활성적인 또는 억제적인 기능을 갖고 있다고 알려져 있는 FGF-7 성장인자, BMP수용체 II, 그리고 TGF-$\beta$ 수용체 II 유전자의 비정상 발현이 폐기포 생성에 관여하는지를 각각의 단일성 클론 항체를 사용하여 수술로 절제된 자연기흉 환자의 폐기포 조직들을 면역조직염색 방법으로 염색하여 관찰하였다. 대상 및 방법: 재발성 또는 지속성 기흉으로 흉강경 또는 개흉술로 폐기포 절제술을 실시한 환자들을 대상으로 하였다. 총 31명의 환자로 15세에서 39세까지 연령분포를 보였으며 남자 30명, 여자 1명이었다. 폐기포 절제는 비디오흉강경이나 소절개개흠술을 통하여 폐기포벽에 손상을 가하지 않게 주의하면서 비디오흉강경용 스태플러(Endo GIA stapler)를 이용하여 절제하였으며 가능한 원형을 유지하여 신선한 상태로 포르마린에 고정하여 면역조직화학적 연구를 위한 표본을 만들었다. 폐기포 조직 슬라이드를 단일클론성 항 TGF-$\beta$ 수용체 II, BMP수용체 II 그리고 FGF-7인자 항체를 이용하여 면역조직학적 염색방법으로 관찰하였다. 결과: 전체 환자 31명중 TGF-$\beta$ 수용체 II항체에 양성 반응을 나타낸 환자수는 모두 24명이었다. 이들 중에는 18명이 강한 양성 반응을 보였고, 6명이 약한 양성 반응을 보였다 면역조직화학적 염색 결과를 고배율 현미경으로 살펴보면, TGF-$\beta$ 수용체 II의 염색이 기흉과 정상 폐조직 경계 부위에서 측히 강하게 염색됨이 관찰되었다. 이에 반하여, BMP수용체 II 그리고 FGF-7인자의 항체를 이용한 면역조직학적 염색 결과는 모든 환자의 조직들에서 음성으로 관찰되었다. 결론: 폐 조직이 형성될 때, 억제유전자의 역할을 담당하고 있다고 알려진 TGF-$\beta$ 수용체 II의 발현이 증가되면서 폐기포가 생성될 수 있다는 가능성을 제시하였다. 이번 결론은 면역조직학적 염색 실험 결과만으로 밝혀진 사실임으로 좀 더 체계적인 분자생물학적 인 연구가 요구된다.

• Journal of Ginseng Research
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• 제41권3호
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• pp.411-418
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• 2017

Background: Recently, protein from ginseng was studied and used for the treatment of several kinds of diseases. However, the effect of ginseng total protein (GTP) on proliferation and wound healing in fibroblast cells remains unclear. Methods: In this study, cell viability was analyzed using the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay. Cell cycle distribution was analyzed by flow cytometer. The levels of transforming growth factor ${\beta}1$, vascular endothelial growth factor, and collagens were analyzed by enzyme-linked immunosorbent assay and immunofluorescence staining. The expressions of cyclin A, phosphorylation of extracellular signal-related kinase (p-ERK1/2), and ERK1/2 were analyzed by Western blotting. Results: Our results showed that GTP promoted cell proliferation and increased the percentage of cells in S phase through the upregulation of cyclin A in NIH/3T3 cells. We also found that GTP induced the secretion of type I collagen, and promoted the expression of other factors that regulate the synthesis of collagen such as transforming growth factor ${\beta}1$ and vascular endothelial growth factor. In addition, the phosphorylation of ERK1/2 at Thr202/Tyr204 was also increased by GTP. Conclusion: Our studies suggest that GTP promoted proliferation and secretion of collagen in NIH/3T3 cells by activating the ERK signal pathway, which shed light on a potential function of GTP in promoting wound healing.

• 대한의생명과학회지
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• 제12권1호
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• pp.9-16
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• 2006

We investigated the effects of 835-MHz electromagnetic field (EMF), one of the most popular communication frequency band in Korean code-division multiple-access (CDMA) mobile phone system, on cellular signal transduction. For this, we examined the change of intracellular calcium $([Ca^{2+}]_i)$, reactive oxygen species (ROS) and F-actin polymerization after exposure to 835-MHz EMF followed by the treatment of agonists in Rat-2 fibroblast cells. Culture cells were pretreated with serum-tree medium and concomitantly exposed to 835-MHz at specific absorption rate (SAR) of 4.0 W/kg for 24 hr in a specialized designed apparatus based on Transverse Electro Magnetics (TEM) wave theory. Intracellular $Ca^{2+}$ responses to lysophosphatidic acid (LPA) and epidermal growth factor (EGF) in Rat-2 fibroblast after exposure to 835-MHz EMF were shown to be similar pattern as observed in normal cultured cells. However, the LPA-induced calcium spiking was slightly delayed to 7 sec and sustained thereafter to a little higher ground level under 835-MHz EMF radiation compared to unexposed cells. ROS production level by LPA in the exposed cells was not different from that in control. Furthermore, LPA induced the production of stress fibers with no significant difference in the exposed and unexposed cells. These results suggest that mobile phone radiation (835-MHz, SAR 4.0 W/kg) may not be directly related to signal transduction in Rat-2 fibroblasts except the slight effect of calcium spiking in LPA-induced cells but remain to be further elucidated for possible indirect intervention.

• 대한한의학회지
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• 제43권1호
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• pp.18-32
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• 2022

Objectives: This study aimed to examine the safety, effects on proliferation of hair papilla cells, and anti-inflammatory and antioxidant mechanisms of Artemisia sieversiana Ehrh. ex Willd. (AS) extract. Methods: Safety tests through purity testing, acute toxicity tests, and repeated toxicity tests were performed using AS extract (ASE) which had been dried for over two years. Cell culture and proliferation tests were conducted; VEGF (vascular endothelial growth factor), bFGF (basic fibroblast growth factor), and EGF (epidermal growth factor) and protein expression analyses were performed for mechanistic evaluation; and inhibitory effects of ASE on the RNA expression of testosterone, 5𝛼-reductase, and aromatase was assessed. The anti-inflammatory and antioxidant efficacy of ASE was confirmed by measuring the levels of nitric oxide, inflammatory mediators (TNF-𝛼 and PGE2), inflammatory cytokines (IL-1𝛽, IL-6, and IL-8), and chemokine MCP-1. Results: The safety of ASE was confirmed. The mechanism of cell proliferation in human hair follicle dermal papilla cells involved the promotion of VEGF, bFGF, and EGF expression. ASE decreased mRNA expression of testosterone, 5𝛼-reductase, and aromatase-1 in a concentration-dependent manner. PGE2 and TNF-𝛼 production by inflammatory mediators was also significantly decreased in a concentration-dependent manner, and inflammatory cytokine and chemokine expression was inhibited. Conclusions: ASE is suggested to promote papillary cell growth at the cellular level, to suppress expression of various enzymes involved in hair cycle and cell death, and to inhibit hair loss through anti-androgen, anti-inflammatory, and antioxidant effects.

• 대한치과교정학회지
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• 제31권6호
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• pp.589-600
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• 2001

이 연구의 목적은 IGF-I의 골모세포에 대한 직접적 작용효과와 IGF-I이 섬유모세포에 작용된 후 섬유모세포에서 유리된 인자를 포함한 조절 배양액이 골모세포에 어떤 영향을 미치는지에 대해 각각 치주인대 섬유모세포와 치은 섬유모세포를 이용하여 알아본 후, 치조골의 반응을 외적자극과의 사이에서 매개하는 치주인대의 주세포군인 섬유모세포를 통한 IGF-I의 골모세포에 대한 영향을 알아보는 것이다. 이를 위해 치주인대 섬유모세포와 치은 섬유모세포를 6-8주된 백서(Sprague-Dawley rat)에서 채집하고, 골모세포는 태생 21일된 동종백서에서 채집하여 기본배양을 한 후, 각 군을 6군으로 분리하여 $1{\times}10^4$/we11, (1ml/well)세포수로 분주한 골모세포 배양접시에 IGF-I을 1,10,100ng/m1로 각각 농도를 달리하여 그 효과를 알아보았다. 각각의 군은 대조군, IGF-I을 직접 투여한 골모세포 배양군, 치주인대 섬유모세포의 조절배양액을 이용한 골모세포 배양군, IGF-I으로 처리한 치주인대 섬유모세포의 조절배양액을 이용한 골모세포 배양군, 치은 섬유모세포의 조절배양액을 이용한 골모세포 배양군, IGF-I으로 처 리한 치은 섬유모세포의 조절배양액을 이용한 골모세포 배양군이다. 조절배양액은 배양 36시간후(IGF-I 처리후 12시 간 배양 포함) 채집하였고, 마지막으로 IGF-I 및 조절배양액으로 처리한 후에 추가 24시간 배양한 후, Alkaline phoaphatase 활성도, Western blot 을 이용한 BMP발현, MTT를 이용한 세포증식, BCA kit을 이용한 총단백질량 측정, Western blot을 이용한 교원질 합성 계측 및 골결절의 생성을 관찰하였다. 본 연구의 결과는 다음과 같다. 1 Alkaline phosphatase활성은 10, 100ng/m1의 IGF-I으로 처리한 군과 치주인대 섬유모세포의 조절배양액을 이용한 군, IGF-I으로 처리한 치주인대 섬유모세포의 조절배양액을 이용한 군에서 대조군보다 더 높게 나타났다. 10, 100ng/ml의 IGF-I으로 처리한 치주인대 섬유모세포의 조절배양액을 이용한 실험군에서 유의성 있게 높게 나타났다. 2. 100ng/m1농도의 IGF-I으로 직접 처리한 군에서 골모세포증식이 유의성 있게 증가하였다. 3. 총단백질량은 IGF-I투여와 상관없이 대조군, 실험군 모두 유사하였다. 4. 모든 실험군에서 BMP2,4가 발현되었고, 대조군과 유의한 차이는 없었다. 이상의 결과에서 IGF-I의 투여여부와는 상관없이 치주인대 섬유모세포가 유리하는 물질이 골모세포의 활성을 증가시키는 것으로 나타났으며, IGF-I은 고농도일때만 유의성있게 골모세포 활성을 촉진함을 알 수 있었다. 따라서 이 연구를 통하여 치주인대 섬유모세포가 골모세포활성을 촉진 시키는 작용을 가지고 있음이 확인되었다.

• 대한치과보철학회지
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• 제43권3호
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• pp.393-408
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• 2005

Statement of problem. In the process of bone formation, titanium (Ti) surface roughness is an important factor modulating osteoblastic function. Purpose. This study was carried out to determine the effect of different Ti surface on biologic responses of a human osteoblast-like cell line (MG63). Materials and methods. MG63 cells were cultured on S (smooth), SLA (sandblasted largegrit & acid etching), HA (hydroxyapatite) Ti. The morphology and attachment of the cells were examined by SEM. The cDNAs prepared from total RNAs of MG63 were hybridized to a human cDNA microarray (1,152 elements). Results. The appearances of the surfaces observed with SEM were different in the three types of dental substrates. The surface of SLA and HA were shown to be rougher than S. MG63 cells cultured on SLA and HA were cell-matrix interaction. In the expression of genes involved in osseointegration, upregulated genes were bone morphogenetic protein, Villin, Integrin, Insulin-like growth factors in different surfaces. Downregulated genes were fibroblast growth factor receptor 4, Bcl 2-related protein, collagen, CD4 in different surfaces. Conclusion. The attachment and expression of key osteogenic regulatory genes were enhanced by surface roughness of the dental materials.

• Journal of Periodontal and Implant Science
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• 제24권2호
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• pp.357-375
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• 1994

The purpose of this study was to investigate the differences of histochemical characteristics in inflammatory fibrous gingival hyperplasia (FGH), phenytoin-induced gingival hyperplasia(PIGH), idiopathic gingival hyperplasia(IDGH) and control groups (healthy and inflammatory gingiva) by immunohistochemical method with various antibodies and histomorphological analysis. In immunohistochemical finding, antibodies to inflammatory cells (T/B lymphocytes, macrophages, other monocytes), proliferating cell nuclear antigen(PCNA), epidermal growth factor(EGF), factor VIII, and type I collagen were used. 1. The inflammatory infiltrates in FGH were less than those in inflammatory gingiva. The composition of inflammatory cells of PIGH was similar with that of FGH. IDGH showed a similar histologic findings with healthy gingival tissue. 2. In FGH, the number of fibroblasts and newly-formed collagen fibers was increased. No significant increase of fibroblasts and the dense accumulation of thick collagen fibers were seen in PIGH. The increase of fibroblasts and the dense accumulation of thick collagen were seen in IDGH. 3. PCNA-positive cells were localized mainly in the area accumulated with inflammatory cells and blood vessels, significantly increased in all hyperplastic tissue groups, and distributed evenly in IDGH. 4. The distribution of EGF were not observed in healthy gingiva but detected locally in area with confluent blood vessels,without significant difference between the other tissue groups. This results suggest that inflammation plays a significant role in inducing hyperplastic change of gingival tissue. While in DIGH, drug itself as well as inflammation seems to attribute to hyperplastic change.

• 대한수의학회지
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• 제38권2호
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• pp.394-400
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• 1998

Cryopreservation of embryos by vitrification is a simple method to preserve bovine embryos for subsequent embryo transfer, but embryonic viability after vitrification has been inconsistent and low compared with conventional slow freezing. The aim of the present study is to examine the effect of serum or serum albumin in a vitrification solution and epidermal growth factor(EGF) or fibroblast growth factor(FGF) on in vitro viability of bovine blastocysts frozen by vitrification. Bovine blastocysts were produced by in vitro maturation, fertilization of follicular oocytes and culture of embryos in a synthetic oviduct fluid medium(SOFM) containing BSA and 19 essential and nonessential amino acids. Blastocysts with excellent or good morphology were selected at 7 or 8 days after culture and utilized for vitrification. In experiment 1, blastocysts were vitrified in a solution containing semi-fetal calf serum(SFCS) or BSA(5 or 10mg/ml) and then their subsequent viabilities were examined by culturing thawed embryos in a SOFM containing BSA and 19 amino acids. Effect of EGF or FGF added to a SOFM containing polyvinyl alcohol(PVA) on the viability of vitrified-thawed blastocysts was investigated in experiment 2. BSA added at 5 or 10mg/ml to a vitrification solution showed significantly higher(p < 0.05) developmental rate to expanded and hatching blastocysts than SFCS, but there was no significant difference in the developmental rate to hatched blastocysts after thawing. Supplementation of a culture medium with EGF and/or FGF significantly increased(p < 0.05) embryo development to expanded blastocysts compared with control but showed no beneficial effect on the development to hatching or hatched blastocysts. Coculture of thawed embryos with granulosa cells in a TCM 199 containing 10% fetal calf serum(FCS) showed the highest developmental rate to expanded, hatching and hatched blastocysts among the groups tested. In conclusion, supplementation of a vitrification solution with BSA at 5mg/ml and culture of thawed blastocysts in a medium containing EGF and/or FGF can improve in vitro viability of bovine blastocysts frozen by vitrification.

• 동의생리병리학회지
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• 제23권4호
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• pp.799-804
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• 2009

Kamikaekyuk-tang(KMKKT), a formula of ten Oriental herbs, was orientally designed to promote vital energy, to remove blood stasis, and to decrease inflammation for treating cancers. KMKKT and its component had potent antiandrogen and androgen receptor activities in prostate cancer and also inhibited angiogenesis induced by basic fibroblast growth factor (bFGF) in human umbilical vein endothelial cells and suppressed the tumor growth in LLC-bearing mice, and liver metastasis of colon 26-L5 cancer cells, suggesting a potent cancer preventive agent. Nevertheless, there is no safety study of KMKKT before clinical trial so far. Thus, in the current study, we investigated the toxicity about ethanol-extracted KMKKT. Male and female Spraque Dawley (SD) rats were given orally by KMKKT at 250, 500, and 1000 mg/kg for 4 weeks. Mortality, clinical signs and measured change of body weight, food consumption and water consumption were observed. In addition, we performed ophthalmologic, urinary, hematological, blood serum biochemical and histopathological examination. Any general toxicity was not found in KMKKT treated group. Also, there were no significant differences in the parameters such as body weight, food consumption and water consumption, a lot of urine and blood factor levels except WBC, MCHC and Ca level compared with control group. Although WBC and MCHC were elevated in female rats and Ca level was decreased in male rats, these were within normal ranges. Finally, we determined that maximum tolerated dose (MTD) was 1000 mg/kg and no observed adverse effect level (NOAEL) was 500 mg/kg. Taken together, these results demonstrated that KMKKT is very safe to SD rats.