• Asian-Australasian Journal of Animal Sciences
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• 제31권5호
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• pp.650-657
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• 2018

Objective: The study investigated the biological functions and mechanisms for controlling cashmere growth of Liaoning cashmere goat by ovarian carcinoma immunoreactive antigen-like protein 2 (OCIAD2) and decorin (DCN) genes. Methods: cDNA library of Liaoning cashmere goat was constructed in early stages. OCIAD2 and DCN genes related to cashmere growth were identified by homology analysis comparison. The expression location of OCIAD2 and DCN genes in primary and secondary hair follicles (SF) was performed using in situ hybridization. The expression of OCIAD2 and DCN genes in primary and SF was performed using real-time polymerase chain reaction (PCR). Results: In situ hybridization revealed that OCIAD2 and DCN were expressed in the inner root sheath of Liaoning cashmere goat hair follicles. Real-time quantitative PCR showed that these genes were highly expressed in SF during anagen, while these genes were highly expressed in primary hair follicle in catagen phase. Melatonin (MT) inhibited the expression of OCIAD2 and promoted the expression of DCN. Insulin-like growth factors-1 (IGF-1) inhibited the expression of OCIAD2 and DCN, while fibroblast growth factors 5 (FGF5) promoted the expression of these genes. MT and IGF-1 promoted OCIAD2 synergistically, while MT and FGF5 inhibited the genes simultaneously. MT+IGF-1/MT+FGF5 inhibited DCN gene. RNAi technology showed that OCIAD2 expression was promoted, while that of DCN was inhibited. Conclusion: Activation of bone morphogenetic protein (BMP) signaling pathway up-regulated OCIAD2 expression and stimulated SF to control cell proliferation. DCN gene affected hair follicle morphogenesis and periodic changes by promoting transforming growth $factor-{\beta}$ ($TGF-{\beta}$) and BMP signaling pathways. OCIAD2 and DCN genes have opposite effects on $TGF-{\beta}$ signaling pathway and inhibit each other to affect the hair growth.

• 생명과학회지
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• 제29권4호
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• pp.499-508
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• 2019

암줄기세포는 전이와 재발의 주요한 요인이 되는 자가재생능력, 분화할 수 있는 능력, 치료에 대한 저항성 및 암 형성 능력의 특성을 가진다. WNT/${\beta}$-catenin, Hedgehog, Notch, BMI1, BMP 및 TGF-${\beta}$와 같은 암줄기세포의 특성을 획득 및 유지할 수 있는 신호기전의 연구 결과가 존재하지만, 현재까지 선택적으로 암줄기세포를 표적할 수 있는 치료 전략은 미미하다. 최근, 면역관문억제제인 CTLA-4, PD-1/PD-L1 단일클론항체는 흑색종, 폐암, 췌장암 및 혈액암에 괄목할만한 임상 시험 결과를 나타냈으며, 긴 항암지속효과와 적은 부작용은 기존 항암제보다 개선 된 모습을 보였다. 또한 두경부편평상피암, 흑색종, 유방암 줄기세포를 선택적으로 제거 하였다. 위의 결과를 종합하면, 면역관문억제제는 이전 항암제에 비해 효과적인 항암전략이며, 동시에 암줄기세포를 선택적으로 제거할 수 있는 가능성을 시사한다. 따라서 본 리뷰에서는 암줄기세포와 면역관문억제제의 이해를 통해, 면역관문억제제의 암줄기세포 표적 가능성에 대해 고찰하고자 한다.

• 한국발생생물학회지:발생과생식
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• 제15권1호
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• pp.41-52
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• 2011

In the present study, embryoid bodies (EBs) obtained from induced pluripotent stem cells (iPSCs) were induced to differentiate into germ lineage cells by treatment with bone morphogenetic protein 4 (BMP4) and retinoic acid (RA). The results were compared to the results for embryonic stem cells (ESCs) and multipotent spermatogonial stem cells (mSSCs) and quantified using immunocytochemical analysis of germ cell-specific markers (integrin-${\alpha}6$, GFR-${\alpha}1$, CD90/Thy1), fluorescence activating cell sorting (FACS), and real time-RT-PCR. We show that the highest levels of germ cell marker-expressing cells were obtained from groups treated with 10 ng/$m{\ell}$ BMP4 or 0.01 ${\mu}M$ RA. In the BMP4-treated group, GFR-${\alpha}1$ and CD90/Thy-1 were highly expressed in the EBs of iPSCs and ESCs compared to EBs of mSSCs. The expression of Nanog was much lower in iPSCs compared to ESCs and mSSCs. In the RA treated group, the level of GFR-${\alpha}1$ and CD90/Thy-1 expression in the EBs of mSSCs Induced pluripotent stem cells, Mouse embryonic stem cells, Multipotent spermatogonial stem cells, Germ cell lineage, Differentiation potential. was much higher than the levels found in the EBs of iPSCs and similar to the levels found in the EBs of ESCs. FACS analysis using integrin-${\alpha}6$, GFR-${\alpha}1$, CD90/Thy1 and immunocytochemistry using GFR-${\alpha}1$ antibody showed similar gene expression results. Therefore our results show that iPSC has the potential to differentiate into germ cells and suggest that a protocol optimizing germ cell induction from iPSC should be developed because of their potential usefulness in clinical applications requiring patient-specific cells.

• 대한치과보철학회지
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• 제49권3호
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• pp.254-262
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• 2011

연구 목적: 기존의 기계절삭 가공된 티타늄 표면에 생체활성 유기물질이 코팅된 생체 활성 임플란트에 관한연구가 최근에 많이 이루어지고 있다. 이러한 생체 활성 임플란트는 임플란트 자체에 골치유 및 골형성 효과를 부여할 수 있다. 본 연구의 목적은 생체 활성 임플란트 표면에 대한 연구 동향을 분석함으로써, 표면처리 기술 개발의 현황과 향후 발전 전망에 대해서 알아보고자 하였다. 연구 재료 및 방법: 본 문헌 분석 연구는 'Web of Science (http://isiknowledge.com, Thomson Scientific)'를 통해 SCI (science citation index) 등재 논문들을 기반으로 분석하였다. 주제별로 연구비중을 비교하기 위해서, 임플란트 표면 코팅에 사용되는 생체활성 유기물들로 키워드를 선정하고 각 키워드로 대표되는 리서치 그룹을 선정하여 각 그룹들의 연구 동향을 분석하였다. 또한 키워드별로 동물실험 동향을 분석하여 상용화 가능성에 대해서도 고찰하였다. 결과: 연구의 비중은 콜라겐, 피브로넥틴, 골형성단백질, RGD (Arg-Gly-Asp) 순으로 나타났으며, 콜라겐을 이용한 연구가 40%로 가장 많은 연구가 이루어지고 있었다. 네가지 주제 모두 시간의 경과에 따라 연구횟수가 증가하는 양상을 보였으며, 특히 RGD에 관한 연구횟수가 가장 급격하게 증가하였다. 콜라겐 대표 리서치 그룹의 연구 동향을 분석하였을 때, 콜라겐은 단독 사용보다는 다른 생체활성 유기물 및 무기물과의 병용 처치에 관해 주로 연구가 되고 있었으며, 특히 세포외기질인 콘드로이틴 설패이트와의 병용 처치에 관한 연구가 가장 많이 이루어지고 있는 것으로 나타났다. 골형성단백질의 대표 리서치 그룹들에서는 rhBMP-2 (recombinant human bone morphogenetic protein-2)에 관한 연구가 주로 이루어지고 있었다. 동물실험에 관한 연구 동향 분석 결과 콜라겐은 차폐막 혹은 약물 전달 매체로서 주로 사용되고 있었고, 골형성단백질은 치조골이식술 혹은 임플란트 표면 코팅에 대한 효과가 연구되고 있었다. 결론: 연구 결과를 주제별로 분석하였을 때 주제별로 일관성 있는 결과를 얻기 어려웠으며, 보다 나은 결과를 얻기 위한 기계적 표면처리와 여러 세포외기질, 성장인자 등의 생체물질의 최상의 조합을 찾기 위한 노력이 진행중이었다. 연구가 초기단계에 머물고 있기 때문에 상용화에 이르기까지는 많은 시간과 노력이 필요할 것이다.

• Molecules and Cells
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• 제37권5호
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• pp.406-411
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• 2014

The initial step of atrioventricular (AV) valve development involves the deposition of extracellular matrix (ECM) components of the endocardial cushion and the endocardialmesenchymal transition. While the appropriately regulated expression of the major ECM components, Versican and Hyaluronan, that form the endocardial cushion is important for heart valve development, the underlying mechanism that regulates ECM gene expression remains unclear. We found that zebrafish crip2 expression is restricted to a subset of cells in the AV canal (AVC) endocardium at 55 hours post-fertilization (hpf). Knockdown of crip2 induced a heart-looping defect in zebrafish embryos, although the development of cardiac chambers appeared to be normal. In the AVC of Crip2-deficient embryos, the expression of both versican a and hyaluronan synthase 2 (has2) was highly upregulated, but the expression of bone morphogenetic protein 4 (bmp4) and T-box 2b (tbx2b) in the myocardium and of notch1b in the endocardium in the AVC did not change. Taken together, these results indicate that crip2 plays an important role in AV valve development by downregulating the expression of ECM components in the endocardial cushion.

• Molecules and Cells
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• 제37권4호
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• pp.330-336
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• 2014

Thymosin beta4 (TB4) has multiple functions in cellular response in processes as diverse as embryonic organ development and the pathogeneses of disease, especially those associated with cardiac coronary vessels. However, the specific roles played by TB4 during heart valve development in vertebrates are largely unknown. Here, we identified a novel function of TB4 in endothelial-mesenchymal transformation (EMT) in cardiac valve endocardial cushions in zebrafish. The expressions of thymosin family members in developing zebrafish embryos were determined by whole mount in situ hybridization. Of the thymosin family members only zTB4 was expressed in the developing heart region. Cardiac valve development at 48 h post fertilization was defected in zebrafish TB4 (zTB4) morpholino-injected embryos (morphants). In zTB4 morphants, abnormal linear heart tube development was observed. The expressions of bone morphogenetic protein (BMP) 4, notch1b, and hyaluronic acid synthase (HAS) 2 genes were also markedly reduced in atrio-ventricular canal (AVC). Endocardial cells in the AVC region were stained with anti-Zn5 antibody reactive against Dm-grasp (an EMT marker) to observe EMT in developing cardiac valves in zTB4 morphants. EMT marker expression in valve endothelial cells was confirmed after transfection with TB4 siRNA in the presence of transforming growth factor ${\beta}$ ($TGF{\beta}$) by RT-PCR and immunofluorescent assay. Zn5-positive endocardial AVC cells were not observed in zTB4 morphants, and knockdown of TB4 suppressed TGF-${\beta}$-induced EMT in ovine valve endothelial cells. Taken together, our results demonstrate that TB4 plays a pivotal role in cardiac valve formation by increasing EMT.

• 한국발생생물학회지:발생과생식
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• 제25권4호
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• pp.279-291
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• 2021

Hair loss is one of the most common chronic diseases, with a detrimental effect on a patient's psychosocial life. Hair loss results from damage to the hair follicle (HF) and/or hair regeneration cycle. Various damaging factors, such as hereditary, inflammation, and aging, impair hair regeneration by inhibiting the activation of hair follicle stem cells (HFSCs) and dermal papilla cells (DPCs). Formyl peptide receptor 2 (FPR2) regulates the inflammatory response and the activity of various types of stem cells, and has recently been reported to have a protective effect on hair loss. Given that stem cell activity is the driving force for hair regeneration, we hypothesized that FPR2 influences hair regeneration by mediating HFSC activity. To prove this hypothesis, we investigated the role of FPR2 in hair regeneration using Fpr2 knockout (KO) mice. Fpr2 KO mice were found to have excessive hair loss and abnormal HF structures and skin layer construction compared to wild-type (WT) mice. The levels of Sonic hedgehog (Shh) and β-catenin, which promote HF regeneration, were significantly decreased, and the expression of bone morphogenetic protein (Bmp)2/4, an inhibitor of the anagen phase, was significantly increased in Fpr2 KO mice compared to WT mice. The proliferation of HFSCs and DPCs was significantly lower in Fpr2 KO mice than in WT mice. These findings demonstrate that FPR2 impacts signaling molecules that regulate HF regeneration, and is involved in the proliferation of HFSCs and DPCs, exerting a protective effect on hair loss.

• Clinical and Experimental Reproductive Medicine
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• 제37권1호
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• pp.13-23
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• 2010

목 적: 본 연구는 인간 배아줄기세포를 생식세포로의 분화를 유도하고 효소에 의해 분리된 단일 배아줄기세포의 배양조건을 확립하기 위해 수행하였다. 연구방법: Embryonic body (EB)는 배아줄기세포 (hESCs) colony를 떼어내어 3일간 hanging drop culture 방법으로 작성하였고, 이러한 EB를 retionic acid (RA)와 bone morphogenetic protein-4 (BMP4)를 단독 또는 함께 배양액에 첨가하여 14일간 배양함으로써 생식세포로의 분화를 유도하였다. 분화를 유도한 EB는 생식세포 발현유전자인 c-kit과 VASA의 표지인자를 이용한 면역조직형광법으로 분화여부를 조사하였다. 줄기세포는 Collagenase, Tryple 그리고 Accutase 등의 효소로 각각 분리하였고 분리된 단일세포들의 colony formation rate를 조사하였다. 한편 Rho-associated kinase inhibitor (Y-27632)를 단일세포 배양액에 첨가하여 단일세포 분리과정 중에 발생하는 apoptotic damage를 감소시키고자 하였다. 결 과: Tryple 또는 Accutase를 이용하여 분리한 단일세포가 Collagenase에 의해 분리된 세포에 비해 높은 colony formation rate를 나타내었다. 단일세포를 $5{\times}10^3$ cells/well (4 well dish) 농도로 지지세포 위에 seeding하였을 때 다른 농도의 세포를 seeding한 것에 비해 높은 colony formation rate를 확보하는데 효과적이었다. Y27632의 첨가는 단일세포의 colony formation rate를 유의하게 향상시켰으며 특히 Tryple로 분리한 단일세포에 보다 효과적이었다. EB의 분화유도후 c-kit과 VASA의 표지인자를 이용한 면역조직형광염색은 대조군인 정소조직에 비해 약한 형광염색을 나타내었다. 결 론: Tryple을 이용한 단일세포 분리가 건강한 단일세포를 얻는데 가장 효율적이었으며 Y27632 의 첨가는 단일 세포의 생존 및 colony formation에 유익하다는 것을 확인하였다. 다른 연구와는 달리 본 연구에서는 단지 생식세포 표지인자의 희미한 형광염색만을 관찰하였는데 이러한 결과는 본 연구의 분화유도기간이 상대적으로 짧았던 것이 원인이었을 것으로 생각된다.

• 한방재활의학과학회지
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• 제28권1호
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• pp.1-17
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• 2018

Objectives The purpose of this study was to evaluate the effect of Jeopgolsan (JGS) extract on anti-oxidant, anti-inflammatory activities in RAW 264.7 cells and on factors related with fracture healing in skull fractured rat. Methods Experimental animals were divided into four groups: normal group without any treatment (Normal), contral group were treated orally with distilled water (Control), Experimental group were treated orally with JGS at a concentration of 200 mg/kg/day (JGS 200) and Experimental group were treated orally with JGS at a concentration of 200 mg/kg/day (JGS 400). Rats in each group except the normal group were induced fractures in the skull. The 1,1-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS) radical scavenging activity were measured to evaluate antioxidant activity. The production of nitric oxide (NO), $interleukin-1{\beta}$ ($IL-1{\beta}$), interleukin-6 (IL-6) and tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$) in the RAW 264.7 cells were measured to evaluate anti-inflammatory activity. The production of osteocalcin calcitonin, carboxy-terminal telepeptides of type II collagen (CTX II), transforming growth $factor-{\beta}$ ($TGF-{\beta}$), bone morphogenetic protein-2 (BMP-2), Insulin and alkaline phosphatase (ALP) in serum of rats were measured to evaluate the effects of fracture healing at 0, 2, 4, and 6th week. X-rays were taken every 3 week from 0 to 6th week to evaluate fracture healing effect. Results 1. No cytotoxicity was observed. 2. DPPH and ABTS radical scavenging activity were increased in a concentration dependent manner, indicating anti-oxidant effect. 3. NO, $IL-1{\beta}$, IL-6, and $TNF-{\alpha}$ were not significantly changed, indicating no anti-inflammatory effect. 4. Osteocalcin, Calcitonin, $TGF-{\beta}$ and ALP were significantly increased in the experimental groups. 5. CTX II, insulin were significantly decreased in the expermental groups. 6. Radiologic examination showed that union of fracture was promoted. Conclusions From above results, JGS showed significant results in factors related with fracture healing and radiologic examination. Threfore, JGS is expected to be effective in the treatment of fracture.