• Natural Product Sciences
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• 제20권4호
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• pp.262-268
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• 2014

Rutaecarpine is one of the major alkaloids present in the fruits of Evodia rutaecarpa. In this study, rutaecarpine was evaluated, both in vitro and in vivo, for its hepatoprotective properties against thioacetamide (TAA)-induced hepatic fibrosis. The results showed that rutaecarpine inhibited TAA-induced cytotoxicity, reduced the expression of the fibrogenic cytokine transforming growth factor ${\beta}1$ ($TGF-{\beta}1$), and induced the expression of bcl-2. To evaluate its in vivo effects, animal models with TAA-induced hepatic fibrosis were utilized. Levels of liver tissue injury-associated enzymes, including alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were monitored. $TGF-{\beta}1$ and the ${\alpha}$-smooth muscle actin (${\alpha}$-SMA) were measured as markers of the protective effects on hepatic fibrosis. The AST and ALT levels in blood were greatly enhanced by TAA and completely blunted by rutaecarpine. Rutaecarpine led to the down-regulation of $TGF-{\beta}$ and Bax mRNA expression, as well as the up-regulation of Bcl-2 and $Bcl-X_L$ mRNA levels. In conclusion, rutaecarpine inhibited TAA-induced hepatic fibrosis and apoptosis by inducing the expression of Bcl-2 while blocking $TGF-{\beta}1$ in our TAA-intoxicated model.

• Journal of Photoscience
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• 제11권3호
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• pp.121-127
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• 2004

The${\beta}$-glucosidases occur widely in all living organisms and has in general a tendency to form oligomers of varying numbers of subunits or aggregates, although the functional implications of such diverse oligomerization schemes remain unclear. In particular, the assembly mode of the oat ${\beta}$-glucosidase is very unique in that it multimerizes by linear stacking of a hexameric building block to form long fibrillar multimers. Some structural proteins such as actin and tubulin assemble into long fibrils in a helical fashion and several enzymes such as GroEL and Pyrodictium ATPase functional complexes, 20S proteasome of the archaebacterium Thermoplasma acidophilum, and lutamine synthetase fromblue-green algae, assemble into discrete oligomers upto 4 stacked rings to maintain their enzymatic activities. In particular, oat ${\beta}$-glucosidase exists in vivo as a discrete long fibrillar multimer assembly that is a novel structure for enzyme protein. It is assembled by linear stacking of hollow trimeric units. The fibril has a long central tunnel connecting to the outer medium via regularly distributed side fenestrations. The enzyme active sites are located within the central tunnel and multimerization increases enzyme affinity to the substrates and catalytic efficiency of the enzyme. Although it is suggested that oligomerization may contribute to the enzyme stability and catalytic efficiency of ${\beta}$-glycosidases, the functional implications of such diverse oligomerization schemes remain unclear so far.

• 한국응용곤충학회지
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• 제35권4호
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• pp.315-320
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• 1996

Drosophila의 actin 5C 유전자 promoter에 쥐의 DNA polymerase $\beta$cDNA를 도입시킨 형질전환 초파리가 고감수성 환경성 변이원 검출계로 사용할 수 있는지를 조사하였다. 체세포 염색체 재조환과 체세포 염색체 돌연변이의 검출을 위해서는 geterozygous(mwh/+) 계통을 사용하였다. 염색체상의 결실이나 비분리 등에 의한 small mwh spot의 자연 발생적 빈도는 non-transgenic w 계통과 transgenic p[pol $\beta$]-130 계통에서 각각 0.351 및 0.606 정도였다. 체세포 염색체 재조환에 의한 large mwh spot의 자연 발생적 빈도의 경우는 transgenic p[pol $\beta$]-130 계통(0.063)이 non-transgenic w 계통(0.021)에 비해 약 3배 정도 높게 나타났다. IQ, Glu-P-1 및 {TEX}$AFB_{1}${/TEX} 등의 돌연변이원의 처리에 의한 경우, 두 종류의 mutant clone의 발생 빈도는 쥐의 DNA polymerase $\beta$가 도입된 transgenic p[pol $\beta$]-130 계통이 non-transgenic w 계통에 비하여 모두 약 2-3배 정도 높게 나타났다. 본 연구의 결과는 쥐의 DNA polymerase $\beta$가 최소한 체세포 염색체 돌연변이 유발이나 체세포 염색체 재조환의 생성 과정에 관여함을 의미하며, 형질전환 초파리 계통이 환경성 변이원 검출계로서 충분한 응용가능성이 있음을 보여 주었다.

• Journal of Periodontal and Implant Science
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• 제37권sup2호
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• pp.447-463
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• 2007

치주인대세포는 시험관적 실험에서 광물화 결절형성을 유도할 수 있으므로 광물화 결절형성에 관여하는 유전자들을 특이하게 발현할 것으로 여겨진다. 이에 본 실험은 cDNA microarray를 이용한 동시 유전자분석을 시행하여 치주인대세포의 분화에 의한 광물화 결절형성시 나타나는 유전자의 특징적 발현 양상을 알아보고자 하였다. 교정치료를 목적으로 경북대학교병원에 내원한 환자의 제일소구치를 발치하여 통상적 방법으로 치주인대세포를 분리, 배양하였고, 3세대의 치주인대세포를 사용하여 실험을 시행하였다. 치주인대세포를 100mm 배양접시에 넣고 배양하여 매 2일 마다 배지를 교환해 주고, 10% FBS만을 투여한 대조군으로, ascorbic acid $(50\;{\mu}g/ml)$, ${\beta}-glycerophosphate$ (10 mM) 및 100 nM dexamethasone을 투여한 군을 실험군으로 하였다. 배양된 치주인대세포에 ascorbic acid, ${\beta}-glycerophosphate$, 그리고 dexamethasone을 투여한 실험군에서 21일째 광물화된 결정을 관찰할 수 있었으나 대조군에서는 관찰할 수 없었다. 3063개의 유전자를 분석한 결과 35개 유전자가 대조군에 비해 2배이상 발현이 증가하였고, 38개 유전자는 2배이상 발현이 감소하였다. 형태학적 검사에서 보여준 바와 같이 광물화 형성과정시 관여하는 JGF-2과 IGFBP2와 같은 유전자가 실험군에서 증가하였으며, 세포골격과 세포외기질 형성에 관여하는 proteogycan 1, fibulin-5, keratin 5, ${\beta}-actin$, ${\alpha}-smooth$ muscle actin, capping protein 등도 발현이 실험군에서 증가하였다. 한편 periostin and S100 calcium-binding protein A4는 대조군에서 오히려 높게 나타나므로 이는 배양된 치주인대세포가 그 자체의 표현형을 유지하고 있음을 보여 주고 있다. 그 외 apoptosis를 유발시키는데 관여하는 Dkk-1와 Nip3는 실험군에서 높게 발현되었고, apoptosis를 억제시키는데 관여하는 Btf와 TAX1BP1는 오히려 낮게 발현됨을 알 수 있으므로 이는 실험군에서 치주인대세포가 골아세포로의 분화되었음을 나타낸다.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2003년도 생물공학의 동향(XIII)
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• pp.750-750
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• 2003

• 생명과학회지
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• 제27권10호
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• pp.1199-1206
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• 2017

종양괴사인자 수용체 일종인 Lymphotoxin ${\beta}$ receptor ($LT{\beta}R$)은 림프 구조와 기관 형성에 중요한 역학을 한다. 우리는 fibroblastic reticular cell (FRC)에서 agonistic $anti-LT{\beta}R$ antibody로 $LT{\beta}R$을 자극하면 stress fiber (SF)에 변화가 생긴다는 것을 알았다. MLCK와 ROCK는 세포에서 SF 형성 기여에 중요한 역할을 한다. 본 연구는 MLCK 저해에 초점을 맞추어 $LT{\beta}R$ 신호 전달은 SF 조절로 항섬유화 효과에 대하여 조사하였다. SF 변화에 대한 $LT{\beta}R$의 기능 조사를 위해 agonistic $anti-LT{\beta}R$ antibody로 처리된 FRC와 세포 추출물을 이용하여 immunoblot, fluorescence assay와 Rho-guanosine diphosphate (GDP)/guanosine triphosphate (GTP) exchange 활성 분석법으로 분석하였다. 세포막과 세포골격 연결자 ezrin은 agonistic $anti-LT{\beta}R$ antibody 처리된 FRC에서 완전히 탈인사화가 유도되었다. Actomysoisn에 의한 SF를 확인하였고 인산화 myosin light chain (p-MLC)인 함께 co-localization 되는 것도 확인하였다. ML7 처리된 FRC에서 agonistic $anti-LT{\beta}R$ antibody 처리된 세포에서 관찰되는 유사한 현상인 SF분해, 세포막 응축과 쇠퇴 현상이 나타났다. ROCK 활성저해는 액틴 골격 변화는 유도하였으나 부분적으로 SF가 세포에 남아 있었다. 반면, ML7에 의한 MLCK저해는 SF를 완전히 분해하였다. 또한, $LT{\beta}R$ 자극은 MLC 인산화를 완전히 억제하였지만, Rho-GDP/GTP exchange 활성변화에서는 감소는 되었으나 활성이 완전히 없어지지는 않았다. 결론적으로 이런 결과들은 FRC에서 $LT{\beta}R$신호전달을 통해 유도되는 SF 조절에는 MLCK가 보다 더 강력한 역할을 한다는 것을 제시하고 있다.

• Asian Pacific Journal of Cancer Prevention
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• 제13권11호
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• pp.5671-5675
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• 2012

Objective: Arrestins act as mediators of G protein-coupled receptor (GPCR) desensitization and trafficking, also actin as a scaffold for many intracellular signaling network. The role that ${\beta}$-arrestin 1 plays in gastric cardiac adenocarcinoma (GCA) and its clinicopathologic significance are untouched. Methods: Fifty patients with gastric cardiac adenocarcinoma were retrospectively enrolled and ${\beta}$-arrestin 1 was detected using immunohistochemistry in tissue samples. Results: Nuclear expression of ${\beta}$-arrestin 1 was observed in 78% of GCA samples (39/50) and cytoplasmic expression in 70% (35/50). ${\beta}$-arrestin 1 could be found in both nucleus and cytoplasm of 54% GCA (27/50) or in either of them in 94% (47/50). ${\beta}$-arrestin 1 protein positivity in well/moderately differentiated carcinomas was significantly higher than that in poorly differentiated carcinomas (P=0.005). We found increased expression of ${\beta}$-arrestin 1 in cytoplasm was correlated with lymph nodal metastasis (P=0.002) and pathological lymph nodal staging (P=0.030). We also found ${\beta}$-arrestin 1 to be over-expressed in glandular epithelia cells of mucinous adenocarcinoma, a tumour type associated with an adverse outcome of gastric cardiac adenocarcinoma (P=0.022). Conclusion: ${\beta}$-arrestin 1 is over-expressed in the nucleus and/or cytoplasm of gastric cardiac adenocarcinoma. However, ${\beta}$-arrestin 1 has no relationship with the prognosis of gastric cardiac adenocarcinoma (P>0.05). Our data imply that ${\beta}$-arrestin 1 in cytoplasm may be involved in differentiation and metastasis of gastric cardiac adenocarcinoma.

• 한국양식학회:학술대회논문집
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• 한국양식학회 2003년도 추계학술발표대회 논문요약집
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• pp.25-25
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• 2003

어류의 번식활동은 뇌-뇌하수체-생식소로 이어지는 내분비기관이 주로 담당하고 있으며, 그 중에서 뇌하수체에서 만들어지는 생식선자극호르몬 (GTH)은 생식소 발달에 매우 중요한 역할을 한다. 한편, GTH는 생식소에서 생성되는 스테로이드호르몬에 의해 유전자의 발현이 조절된다. 본 연구에서는 산천어 (Oncorhynchus masou)의 GTH 유전자발현에 미치는 estradiol-17$\beta$ (E2)의 영향을 생체내에서 먼저 조사하고, 유전자발현조절의 메카니즘을 구명하기 위하여 에스트로젠 수용체 (ER)의 cDNA를 cloning하였다. 또한 내분비교란물질로 알려져 있으며 에스트로젠 수용체와의 유사결합작용이 알려진 노닐페놀 (NP)의 영향도 아울러 조사하였다. 미성숙 산천어를 E2 및 NP (각각 5 mg/kg 및 10 mg/kg 체중)로 처리하면 72 시간째에 뇌하수체내의 $\beta$-actin mRNA에 비해 GTH (FSH$\beta$, LH$\beta$) mRNA가 상대적으로 증가하는 경향을 나타내었다. 뇌하수체에서 분리한 ER의 기본적인 구조는 DNA결합도메인 (Zf-C4), 호르몬결합도메인 (hormone_rec), 기타 전사활성화도메인으로 구성되어 있었으며 (그림 1), 기존에 보고된 에스트로젠 수용체중에서 무지개송어 및 대서양연어의 ER과 각각 98%, 96%의 높은 상동성이 관찰되었다. 향후, 준비된 ER을 재료로 하여 GTH 유전자발현의 호르몬의존성을 조사할 예정이다.

• Fisheries and Aquatic Sciences
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• 제15권1호
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• pp.43-47
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• 2012

Growth hormone (GH) transgenesis in fish has the potential to improve aquaculture efficiency and capacity. However, many fast-growing transgenic fish have experienced side effects caused by excess GH expression. To overcome this unwanted issue associated with several GH transgenic mud loach Misgurnus mizolepis lines carrying GH construct driven by a strong ${\beta}$-actin regulator ($pml{\beta}$-actGH), we performed an alternative version of GH autotransgenesis using a weaker but more stable regulator, the mud loach lectin promoter. GH transgenesis with a pmlectGH construct consisting of the mud loach GH gene driven by the 2.3-kb lectin promoter exhibited significant growth stimulation. However, the extent of the growth acceleration in pmlectGH transgenics (six times maximum when assessed 2 months post hatching) was much less than that in transgenic individuals carrying the $pml{\beta}$-actGH construct. Additionally, the extraordinary gigantism that was common in $pml{\beta}$-actGH-transgenic mud loaches was diminished in transgenic loaches harboring the pmlectGH construct. Transgenic founders (pmlectGH) successfully transmitted their transgene into the next generation with up to 41% frequency. Growth stimulation also persisted in the transgenic F1 strains, with a seven-fold increase in maximum body weight at 6 months of age.

• Journal of Chest Surgery
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• 제44권6호
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• pp.406-412
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• 2011

Background: Development of thoracic aortic aneurysms and aortic dissections (TAAD) is attributed to unbearable wall tension superimposed on defective aortic wall integrity and impaired aortic repair mechanisms. Central to this repair mechanisms are well-balanced and adequately functional cellular components of the aortic wall, including endothelial cells, smooth muscle cells (SMCs), inflammatory cells, and adventitial fibroblasts. Adventitial fibroblasts naturally produce aortic extracellular matrix (ECM), and, when aortic wall is injured, they can be transformed into SMCs, which in turn are involved in aortic remodeling. We postulated the hypothesis that adventitial fibroblasts in patients with TAAD may have defects in ECM production and SMC transformation. Materials and Methods: Adventitial fibroblasts were procured from the adventitial layer of fresh aortic tissues of patients with TAAD (Group I) and of multi-organ donors (Group II), and 4-passage cell culture was performed prior to the experiment. To assess ECM production, cells were treated with TNF-${\alpha}$ (50 pM) and the expression of MMP-2/MMP-3 was analyzed using western blot technique. To assess SMC transformation capacity, cells were treated with TGF-${\beta}1$ and expression of SM ${\alpha}$-actin, SM-MHC, Ki-67 and SM calponin was evaluated using western blot technique. Fibroblasts were then treated with TGF-${\beta}1$ (10 pM) for up to 10 days with TGF-${\beta}1$ supplementation every 2 days, and the proportion of transformed SMC in the cell line was measured using immunofluorescence assay for fibroblast surface antigen every 2 days. Results: MMP-3 expression was significantly lower in group I than in group II. TGF-${\beta}1$-stimulated adventitial fibroblasts in group I expressed less SM ${\alpha}$-actin, SM-MHC, and Ki-67 than in group II. SM-calponin expression was not different between the two groups. Presence of fibroblast was observed on immunofluorescence assay after more than 6 days of TGF-${\beta}1$ treatment in group I, while most fibroblasts were transformed to SMC within 4 days in group II. Conclusion: ECM production and SMC transformation are compromised in adventitial fibroblasts from patients with TAAD. This result suggests that functional restoration of adventitial fibroblasts could well be a novel approach for the prevention and treatment of TAAD.