• Asian-Australasian Journal of Animal Sciences
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• 제33권11호
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• pp.1837-1847
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• 2020

Objective: To evaluate the pancreatic differentiation potential of α-1,3-galactosyltransferase knockout (GalTKO) pig-derived bone marrow-derived mesenchymal stem cells (BM-MSCs) using epigenetic modifiers with different pancreatic induction media. Methods: The BM-MSCs have been differentiated into pancreatic β-like cells by inducing the overexpression of key transcription regulatory factors or by exposure to specific soluble inducers/small molecules. In this study, we evaluated the pancreatic differentiation of GalTKO pig-derived BM-MSCs using epigenetic modifiers, 5-azacytidine (5-Aza) and valproic acid (VPA), and two types of pancreatic induction media - advanced Dulbecco's modified Eagle's medium (ADMEM)-based and N2B27-based media. GalTKO BM-MSCs were treated with pancreatic induction media and the expression of pancreas-islets-specific markers was evaluated by real-time quantitative polymerase chain reaction, Western blotting, and immunofluorescence. Morphological changes and changes in the 5'-C-phosphate-G-3' (CpG) island methylation patterns were also evaluated. Results: The expression of the pluripotent marker (POU class 5 homeobox 1 [OCT4]) was upregulated upon exposure to 5-Aza and/or VPA. GalTKO BM-MSCs showed increased expression of neurogenic differentiation 1 in the ADMEM-based (5-Aza) media, while the expression of NK6 homeobox 1 was elevated in cells induced with the N2B27-based (5-Aza) media. Moreover, the morphological transition and formation of islets-like cellular clusters were also prominent in the cells induced with the N2B27-based media with 5-Aza. The higher insulin expression revealed the augmented trans-differentiation ability of GalTKO BM-MSCs into pancreatic β-like cells in the N2B27-based media than in the ADMEM-based media. Conclusion: 5-Aza treated GalTKO BM-MSCs showed an enhanced demethylation pattern in the second CpG island of the OCT4 promoter region compared to that in the GalTKO BM-MSCs. The exposure of GalTKO pig-derived BM-MSCs to the N2B27-based microenvironment can significantly enhance their trans-differentiation ability into pancreatic β-like cells.

• 생명과학회지
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• 제33권11호
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• pp.851-858
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• 2023

Unique cartilage matrix-associated protein (UCMA)은 γ-카르복실화(Gla) 잔기가 풍부한 간외 비타민 K 의존 단백질이다. UCMA는 조골세포 분화를 촉진하고 뼈 형성을 강화한다고 보고되고 있지만 고혈당 스트레스 하에서 조골세포에 미치는 영향에 대해서는 아직 알려진 바가 없다. 본 연구에서는 고혈당 조건하에서의 MC3T3-E1 조골세포에서 UCMA 효과를 조사하기 위해 MC3T3-E1 조골세포를 높은 포도당에 노출한 후 재조합 UCMA 단백질을 처리하였다. MC3T3-E1 세포에서 활성 산소종(ROS)의 생성은 고혈당 조건하에서 증가했으나 UCMA 단백질 처리 후 감소했음을 CellROX 및 MitoSOX 염색으로 확인하였다. 또한 고혈당 조건에서 UCMA 단백질을 함께 처리한 MC3T3-E1 세포에서 정량적 중합효소 연쇄반응 결과, 항산화 유전자인 nuclear factor erythroid 2-related factor 2 와 superoxide dismutase 1 발현이 증가하였다. 동일 조건하에서 UCMA 단백질 처리에 의해 heme oxygenase-1 발현 감소와 함께 세포질에서 핵으로의 전위가 감소되었고, 미토콘드리아 분열에 관여하는 dynamin-related protein 1 발현이 증가하였으며, AKT 신호 활성은 억제되었다. 종합적으로 UCMA는 고혈당에 노출된 조골세포에서 ROS 생성을 완화하고, 항산화 유전자 발현을 증가시키고, 미토콘드리아 역학에 영향을 미치며, AKT 신호를 조절하는 것으로 보인다. 본 연구는 UCMA의 세포 메커니즘에 대한 이해를 돕고, 대사 장애 관련한 골 합병증에 대한 새로운 치료제로서의 잠재적 사용 가능성을 제시하고 있다.

• Clinical and Experimental Pediatrics
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• 제56권6호
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• pp.247-253
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• 2013

Purpose: In children with acute leukemia, bone marrow genetic abnormalities (GA) have prognostic significance, and may be the basis for minimal residual disease monitoring. Since April 2007, we have used a multiplex reverse transcriptase-polymerase chain reaction tool (HemaVision) to detect of GA. Methods: In this study, we reviewed the results of HemaVision screening in 270 children with acute leukemia, newly diagnosed at The Catholic University of Korea from April 2007 to December 2011, and compared the results with those of fluorescence in situ hybridization (FISH), and G-band karyotyping. Results: Among the 270 children (153 males, 117 females), 187 acute lymphoblastic leukemia and 74 acute myeloid leukemia patients were identified. Overall, GA was detected in 230 patients (85.2%). HemaVision, FISH, and G-band karyotyping identified GA in 125 (46.3%), 126 (46.7%), and 215 patients (79.6%), respectively. TEL-AML1 (20.9%, 39/187) and AML1-ETO (27%, 20/74) were the most common GA in ALL and AML, respectively. Overall sensitivity of HemaVision was 98.4%, with false-negative results in 2 instances: 1 each for TEL-AML1 and MLL-AF4. An aggregate of diseases-specific FISH showed 100% sensitivity in detection of GA covered by HemaVision for actual probes utilized. G-band karyotype revealed GA other than those covered by HemaVison screening in 133 patients (49.3%). Except for hyperdiplody and hypodiploidy, recurrent GA as defined by the World Health Organizationthat were not screened by HemaVision, were absent in the karyotype. Conclusion: HemaVision, supported by an aggregate of FISH tests for important translocations, may allow for accurate diagnosis of GA in Korean children with acute leukemia.

• 대한소아치과학회지
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• 제35권1호
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• pp.30-38
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• 2008

치아 치수 세포는 치아 손상에 따르는 병리적인 상황에서 골과 상아질 기질을 형성하는 능력을 가진 것으로 생각된다. 본 연구에서는 MTA가 사람 치수세포의 성장에 미치는 영향과 상아질 형성에 관여하는 유전자의 발현을 유도하는지를 알아보고자 하였다. 또한 상아질 형성의 잠재적 지표인 alkaline phosphatase(ALP) activity에 미치는 영향을 평가하였다. 유전자 발현 검사를 위해 glyceraldehyde-3-phosphate dehydrogenase, type I collagen, alkaline phosphatase, osteonectin(SPARC), and dentin sialoprotein primer set을 이용하여 MTA 처리 2일과 4일 후 reverse transcriptase polymerase chain reaction(RT-PCR)을 시행하였다. cell viability assay(세포 생존력 측정) 에서 5일간 MTA에 노출된 치수 세포의 비율이 대조군보다 높았다. 대조군에 비해 MTA를 처리한 군에서 ALP와 SPARC가 증가되었다. 이상의 결과를 종합하여 보면, 이 연구에 사용한 dental pulp culture system은 MTA를 포함한 치과재료의 처리 후 치수세포의 성장과 분화 그리고 상아질 형성 유도 기전을 연구하는 데 유용한 모델로 사용할 수 있다. MTA 처리는 사람 치수세포에 세포독성을 유도하지 않으며, ALP 활성도와 유전자 발현 그리고 osteonectin (SPARC) 유전자 발현을 증가시켜 수복상아질을 형성할 것으로 사료된다.

• 한국발생생물학회지:발생과생식
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• 제13권4호
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• pp.249-256
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• 2009

구조적으로 estrogen 수용체(estrogen receptor, ER)와 유사한 estrogen receptor-related receptor(ERR)는 포유동물에서 배발생 후기에 외배엽 형성과 관련되어 있다고 알려진 고아핵수용체(orphan nuclear receptor)이다. ERR은 ER과 DNA binding domain의 보존성은 유사하지만, ligand 결합 및 전사 활성은 다르다. 포유동물의 ERR에 관한 연구에 비하여 해양 무척추동물의 ERR에 대한 기능 연구는 매우 부족하다. 본 연구에서는 우리나라 동해안에 주로 서식하는 둥근성게(Strongylocentrotus nudus) ERR의 초기 발생기 유전자 발현 변화와 전사 활성 기능을 조사하기 위해 먼저 polymerase chain reaction(PCR)을 이용하여 cDNA를 동정하였다. 둥근성게 ERR은 S. purpuratus와 91%의 높은 상동성을 보였으며, 계통수 분석을 통해 무척추동물 ERR의 clade에 포함되는 것을 알았다. 둥근성게 배발생 시기에 ERR 유전자 발현을 알아보기 위하여 real-time PCR을 실시한 결과, 4~64세포기와 유생기에 mRNA level이 높은 경향을 나타내었다. 또한 호르몬 및 co-regulator에 의한 둥근성게 ERR의 전사 활성을 조사한 결과, 호르몬에 의한 특이성은 확인되지 않았지만, peroxisome proliferator activated receptor-(PPAR) $\gamma$ coactivator $1\alpha$(PGC-$1\alpha$)가 둥근성게 ERR의 coactivator임을 입증할 수 있었다. 이 연구 결과는 향후 새로운 ligand 발굴과 coregulator와 의 상호작용 연구에 도움이 될 것으로 사료된다.

• Journal of Acupuncture Research
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• 제30권4호
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• pp.95-105
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• 2013

Objectives : The objective of this study was to investigate the effects of Curcuma longa $L_{INNE}$ pharmacopuncture at $ST_{36}$ on Complete Freund's Adjuvant(CFA)-induced arthritis in rats. Materials and methods : Arthritis was induced by injecting CFA subcutaneously into the left knee joint and paw, and Curcuma longa $L_{INNE}$ pharmacopuncture(CLL-A. $0.0343{\mu}g/kg$; CLL-B. $0.171{\mu}g/kg$; CLL-C. $0.343{\mu}g/kg$) was injected at $ST_{36}$ each other day for 5 times beginning on day 10 after the CFA injection. Paw edema, withdrawal response, hematological, serological and histological observation were assessed. Results : In paw edema volume all 3 groups(CLL-A, CLL-B, CLL-C) showed significant decrease compared to the CFA control group. In withdrawal response to reaction time and withdrawal response to force all 3 groups(CLL-A, CLL-B, CLL-C) showed significant increase compared to the CFA control group. In serum AST, group CLL-C showed significant decrease compared to the CFA control group. In histological observations, in all 3 groups, more normal chondrocytes were observed compared to the CFA control group and safranin O stain showed high positive reaction in the cartilage tissue close to the bone tissue. Conclusions : The results suggest that Curcuma longa $L_{INNE}$ Pharmacopuncture at $ST_{36}$ has a suppressing inflammation effect on Freund's adjuvant arthritis in rats.

• Journal of Periodontal and Implant Science
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• 제34권3호
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• pp.607-622
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• 2004

Recently epidemiologic studies have indicated that the patients with periodontitis may have increased risk of ischemic cardiovascular events, and have suggested the important roles of blood cytokines and acute reactant proteins in the systemic infection and inflammatory response. Periodontitis and coronary heart disease (CHD) may share the common risk factors and the genetic mechanism associated with interleukin(IL)-1A, B and RA genotype may be involved in the production of IL-1. This study was aimed to investigate the relationship between angiographically defined CHD and periodontitis as chronic Gram-negative bacterial infection and to determine whether the IL-1 gene polymorphism is associated in both diseases. Patients under the age of 60 who had undergone diagnostic coronary angiography were enrolled in this study. Subjects were classified as positive CHD (+CHD, n=37) with coronary artery stenosis more than 50% in at least one of major epicardial arteries, and negative CHD (-CHD, n=30) without significant stenosis. After recording the number of missing teeth, periodontal disease severity was measured by means of plaque index (PI), gingival index (GI), bleeding on probing (BOP), probing depth (PD), clinical attachment level (CAL), and radiographic bone loss around all remaining teeth. Gingival crevicular fluid (GCF) was collected from the 4 deepest periodontal pockets and assessed for cytokine ($IL-1{\beta}$, IL-6, IL-1ra, tumor necrosis $factor-{\alpha}$, and prostaglandin $E_2$). Additionally, blood CHD markers, lipid profile, and blood cytokines were analyzed. IL-1 gene cluster genotyping was performed by polymerase chain reaction and enzyme restriction using genomic DNA from buccal swab, and allele 2 frequencies of IL-1A(+4845), IL-1B(+3954), IL-B(-511), and IL-1RA(intron 2) were compared between groups. Even though there was no significant difference in the periodontal parameters between 2 groups, GCF level of $PGE_2$ was significantly higher in the +CHD group(p<0.05). Correlation analysis showed the positive relationship among PD, CAL and coronary artery stenosis(%) and blood $PGE_2$. There was also significant positive relationship between the periodontal parameters (PI, PD, CAL) and the blood CHD markers (leukocyte count, C-reactive protein, and lactic dehyrogenase). IL-1 gene genotyping showed that IL-1A(+3954) allele 2 frequency was significantly higher in the +CHD group compared with the -CHD group (15% vs. 3.3%, OR 5.118,p=0.043). These results suggested that periodontal inflammation is related to systemic blood cytokine and CHD markers, and contributes to cardiovascular disease via systemic inflammatory reaction. IL-1 gene polymorphism might have an influence on periodontal and coronary heart diseases in Korean patients.

• 한국운동역학회지
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• 제28권2호
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• pp.101-108
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• 2018

Objective: The aim of this study was to investigate gender differences in ground reaction force (GRF) components among different speeds of running. Method: Twenty men ($age=22.4{\pm}1.6years$, $mass=73.4{\pm}8.4kg$, $height=176.2{\pm}5.6cm$) and twenty women ($age=20.7{\pm}1.2years$, $mass=55.0{\pm}8.2kg$, $height=163.9{\pm}5.3cm$) participated in this study. All participants were asked to run on an instrumented dual belt treadmill (Bertec, USA) at 8, 12, and 16 km/h for 3 min, after warming up. GRF data were collected from 30 strides while they were running. Hypotheses were tested using one-way ANOVA, and level of significance was set at p-value <.05. Results: The time to passive peaks was significantly earlier in women than in men at three different running speeds (p<.05). Further, the impact loading rates were significantly greater in women than in men at three different running speeds (p<.05). Moreover, the propulsive peak at 8 km/h, which is the slowest running speed, was significantly greater in women than in men (p<.05), and the vertical impulse at 16 km/h, which is the fastest running speed, was significantly greater in men than in women (p<.05). The absolute anteroposterior impulse at 8 km/h was significantly greater in women than in men (p<.05). In addition, as the running speed increased, impact peak, active peak, impact loading rate, breaking peak, propulsive peak, and anteroposterior impulse were significantly increased, but vertical impulse was significantly decreased (p<.05). Conclusion: The impact loading rate is greater in women than in men regardless of different running speeds. Therefore, female runners might be exposed to the risk of potential injuries related to the bone and ligament. Moreover, increased running speeds could lead to higher possibility of running injuries.

• 대한소아치과학회지
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• 제48권3호
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• pp.291-301
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• 2021

이 연구의 목적은 발거 된 매복 상악 과잉치에서 얻은 치수유래 줄기세포의 초기 계대와 후기 계대의 상아질모세포 유전자의 특성을 알아보는 것이다. 전신 의과 병력이 없는 6 - 9세 사이의 남녀아이 12명에게서 서면동의를 얻고 모두 상악에 위치한 과잉치를 발거하여 당일 발거된 과잉치의 치수세포를 채취하였다. 12개의 세포를 각각 3계대와 10계대에서 골형성 유도 분화제를 처리한 군과 처리하지 않은 군을 나누어 실시간 중합효소 연쇄반응을 시행하여 상아질모세포의 특성을 알아보았다. 사용된 유전자는 osteonectin (ONT), alkaline phosphatase (ALP), osteocalcin (OCN), dentin matrix protein 1 (DMP-1), 그리고 dentin sialophosphoprotein (DSPP)였다. 유전자 발현양은, 분화제를 처리하지 않은 군 3계대에서는 ONT, ALP, OCN, DMP-1, DSPP순서로 많이 발현하였다. 분화제를 처리하지 않은 군 10계대에서는 ONT, DMP-1, OCN, ALP, DSPP순으로 ONT, OCN, DSPP의 순서에는 변화가 없지만 ALP, DMP-1의 순서는 서로 바뀌었다. 이상의 결과를 종합해 볼 때, ALP와 DMP-1은 3계대와 10계대 세포의 분화를 위한 중요한 표지자로 사용될 수 있다. 과잉치 치수유래 줄기세포는 상아질모세포의 특성을 가지며, 또한 과잉치가 어린 나이에 발거되고 10계대까지 소요되는 시간이 적게 걸린다는 것을 고려하면, 과잉치는 치아 유래 줄기세포의 공여부로서 훌륭한 활용가능성이 있음을 확인하였다.

• Toxicological Research
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• 제18권1호
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• pp.79-85
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• 2002

YHB216 is one of new recombinant human erythropoietins (rHu-EPO) developed by Yuhan Research Institute. The rHu-EPO products are widely being used for the treatment of various types of anemia. As a series of safety studies on YHB216, we performed the local irritation test (dermal & ocular application) in male New Zealand White rabbits and micronucleus test in male ICR mice. In the skin irritation test, 0.5 ml of YHB216 10,000 IU/ml solution was applied to the back skin of rabbits for 24 hours and sub-sequent observation was performed. There was no induced response after the treatment and the primary irritation index (P.I.I.) was‘0’. In the eye irritation test, 0.1 ml of YHB216 10,000 IU/mL solution was instilled into the conjunctiva of the eye. No treatment-related reaction was observed at the cornea, iris, and conjunctiva. In the micronucleus test, YHB216 was administered intravenously to male mice (6 mice per group) at dose levels of 0, 6,250, 12,500, and 25,000 IU/kg. Bone marrow cells were collected at 24 hours after the treatment. YHB216 treated groups showed no significant difference in the P/N (polychromatic erythrocyte/ normochromatic erythrocyte) ratio and in the number of micronucleated polychromatic erythrocyte com-pared with the control. In conclusion, YHB216 was found to be a non-irritating material up to 10,000 IU/ml in the local irritation test and to be a non-mutagen up to 25,000 IU/kg in the micronucleus test.