• Molecules and Cells
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• v.44 no.10
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• pp.723-735
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• 2021

Spemann organizer is a center of dorsal mesoderm and itself retains the mesoderm character, but it has a stimulatory role for neighboring ectoderm cells in becoming neuroectoderm in gastrula embryos. Goosecoid (Gsc) overexpression in ventral region promotes secondary axis formation including neural tissues, but the role of gsc in neural specification could be indirect. We examined the neural inhibitory and stimulatory roles of gsc in the same cell and neighboring cells contexts. In the animal cap explant system, Gsc overexpression inhibited expression of neural specific genes including foxd4l1.1, zic3, ncam, and neurod. Genome-wide chromatin immunoprecipitation sequencing (ChIP-seq) and promoter analysis of early neural genes of foxd4l1.1 and zic3 were performed to show that the neural inhibitory mode of gsc was direct. Site-directed mutagenesis and serially deleted construct studies of foxd4l1.1 promoter revealed that Gsc directly binds within the foxd4l1.1 promoter to repress its expression. Conjugation assay of animal cap explants was also performed to demonstrate an indirect neural stimulatory role for gsc. The genes for secretory molecules, Chordin and Noggin, were up-regulated in gsc injected cells with the neural fate only achieved in gsc uninjected neighboring cells. These experiments suggested that gsc regulates neuroectoderm formation negatively when expressed in the same cell and positively in neighboring cells via soluble factors. One is a direct suppressive circuit of neural genes in gsc expressing mesoderm cells and the other is an indirect stimulatory circuit for neurogenesis in neighboring ectoderm cells via secreted BMP antagonizers.

• Development and Reproduction
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• v.14 no.4
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• pp.233-241
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• 2010

Human adipose stem cells are an abundant, readily available population of multipotent progenitor cells that reside in adipose tissue and these cells have characteristics very similar to bone marrow mesenchymal stromal cells (BMMSCs). However, liposuction procedure, donor age, body mass index, and harvesting sites might generate differences in the initial cell population and the preparations are a heterogeneous mixture of precursors with different subsets. Therefore, in this study, we investigated the characteristics of human thigh adipose stem cells and the differentiation potential into mesodermal and endodermal lineage. Thigh adipose stem cells maintained fibroblast-like morphology similar to BM-MSCs and they underwent average 56.5 doublings and produced $5{\times}10^{22}$ cells. These cells expressed SCF, Oct4, nanog, vimentin, CK18, FGF5, NCAM, Pax6, BMP4, HNF4a, nestin, GATA4, HLA-ABC, and HLA-DR genes at p3 and they also expressed Oct4, Thy-1, FSP, vWF, vimentin, desmin, CK18, CD54, CD4, CD106, CD31, a-SMA, HLA-ABC proteins. Moreover, they could differentiate into mesodermal lineage cells such as adipocyte, osteoblast and chondrocyte. In addition, they also differentiated into insulin secreting cells in our culture condition. In conclusion, human thigh adipose stem cells retain proliferative potential and expression patterns similar to BM-MSCs and they also differentiate into various cell types. Thus, human thigh adipose stem cells might be useful alternative cell source for clinical application.

• Clinical and Experimental Reproductive Medicine
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• v.36 no.1
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• pp.23-34
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• 2009

Objectives: Mesenchymal stem cells (MSC) comprise a promising tool for cellular therapy. It is known that long-term in vitro culture of human bone marrow and adipose tissue derived-MSCs lead to a reduction of life span and a change of stem-like characters. The aim of our study was to examine whether stem cell properties of human umbilical cord-derived stem cells (HUC) could be affected by in vitro expansion. Methods: HUC were isolated from human umbilical cord and cultured for 10 passages in vitro. Morphology and population doubling time (PDT) were investigated, and changes of stem cell properties were examined using RT-PCR and immunocytochemistry during serial subcultures. Results: Morphology and PDT of HUC began to change slightly from the 7th passage (p7). Expression level of nestin and vimentin mRNAs increased along with the culture period from p4 until p10. In contrast, expression level of SCF mRNA decreased during the same culture period. Expression level of Oct-4 and HNF-4${\alpha}$ mRNAs was not significantly changed throughout the culture period until p10. Expression level of BMP-4, FGF-5, NCAM and HLA-ABC mRNAs appeared to increase as the culture continued, however, the difference was not significant. Immunocytochemical studies showed that HUC at p3, p6 and p9 positively were stained with antibodies against SSEA-3 and SSEA-4 proteins. Interestingly, staining intensity of HUC for ICAM-1 and HLA-ABC gradually increased throughout the culture period. Intensity against thy-1 and fibronectin antibodies increased at p9 while that against TRA-1-60 and VCAM-1 antibodies began to decrease at p6 until p9. Conclusions: These results suggest that HUC change some of their stem cell characteristics during in vitro culture. Development of culture system might be needed for the maintenance of characteristics.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.29 no.3
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• pp.74-94
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• 2016

Objectives : YangHyulEum Gami-Bang(YHEG) is a hair care extracts which is composed of fourteen plant extracts used in oriental medicine. The purpose of this study is to investigate the effect of YangHyulEum Gami-Bang(YHEG) on the alopecia and hair growth.Methods & Results : The herbal extracts from YangHyulEum Gami-Bang(YHEG) was tested using in vivo and in vitro test models. 1. The YHEG extracts showed effect on the DNA proliferation of the hair dermal papilla cells measured by [3H]thymidine incorporation. 2. YHEG showed promoting on the expression of growth factors such as IGF-1, KGF-1 and inhibiting on the expression of inhibitory hair growth factor such as TGF-β1, BMP-2 estimated by qPCR. 3. The YHEG extracts showed effect on the activation of β-catenin in the dermal papilla cells. 4. YHEG showed inhibitory effects of NO synthesis at 0.2% concentrations. 5. YHEG showed effects in the expression of IL-1β, TNF-α, IL-6, COX-2 and iNOS gene in the LPS stimulated RAW 264.7 cells. 6. The hair growth index of the YHEG extracts ranked at over 2 when compared to control group which was ranked at 0. 7. The hair follicle number, length and size of the experimental group were remarkably higher than the control group in the histological observation.Conclusions : These results suggest that YangHyulEum Gami-Bang(YHEG) has hair growth promoting activity and it can be used as a potent treatment agent for preventing hair loss and stimulating hair growth for treatment of alopecia.

• Journal of Korean Medicine Rehabilitation
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• v.26 no.3
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• pp.31-49
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• 2016

Objectives The study was designed to evaluate the healing effects of Joaguihwan (JGH) extract on Anti-oxidation, Anti-inflammatory in RAW 264.7 Cells and factors related with bone metabolism in skull fractured Rat. Methods The fracture healing effect of JGH was measured by scavenging activities of1,1-diphenyl-2-picryl-hydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS) and nitric oxide (NO) in RAW 264.7 cells. The inhibitory effect against the production of inflammatory mediators including interleukin-$1{\beta}$ (IL-$1{\beta}$), interleukin-6 (IL-6), tumor necosis factors-${\alpha}$ (TNF-${\alpha}$) expression was inhibited in RAW 264.7 cells was experimented using JGH. The effects of JGH on healing fractured rats was measured by osteocalcin, calcitonin, CTXII, TGF-${\beta}$, BMP-2, Insulin, ALP in the serum. and was checked every 3 weeks from 0 week to 6week using x-ray. Results 1. DPPH free radica and ABTS scavenging activity of JGH were increased according to concentration of JGH in RAW 264.7 Cells. 2. In the experiment, NO, IL-$1{\beta}$, IL-6, TNF-${\alpha}$ all showed decrease, in general. Especially NO and IL-$1{\beta}$ showed significantly decrease at a concentration of 10, 100 (${\mu}g/ml$). 3. In the production of osteocalcin in the serum, JGH 200, 400 mg/kg experimental group showed significant increased effect at 2 weeks. 4. In the production of calcitonin in the serum. JGH 200 mg/kg experimental group showed significant increased effect at 4, 6 weeks. JGH 400 mg/kg experimental group showed significant increased effect at 2, 4, 6 weeks. 5. In the production of CTX, TGF-${\beta}$, BMP-2 in the serum, experimental group showed increased effect. but no significant effect. 6. In the production of insulin in the serum. JGH 200, 400 mg/kg experimental group showed significant decrease effect at 2, 4, 6 weeks. 7. In the production of ALP in the serum. JGH 200 mg/kg experimental group showed significant increased effect at 2, 4, 6 weeks. JGH 400 mg/kg experimental group showed significant increased effect at 4, 6 weeks. 8. In the change of X-ray, the experimental group showed better healing effects on skull fractured rats than control group. Conclusions From above results, JGH showed healing effect on Anti-oxidation, Anti-inflammatory in RAW 264.7 Cells, factors related with bone metabolism in the serum of skull fractured rat and x-ray, which is expected to be applied in clinics.

• The Korea Journal of Herbology
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• v.36 no.3
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• pp.39-46
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• 2021

Objectives : Mangifera indica leaves are well known for having a variety of benefits, including anti-inflammatory, anti-tumor, diabetic retinopathy and diabetic vasculosis. However, the effects of Mangifera indica leaves on hair loss inhibition have not been studied. In this study, we investigated to find out the activity of Mangifera indica leaves on hair loss. Methods : 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid(ABTS) analysis was performed to confirm the antioxidant efficacy of the water extract of Mangifera indica leaves (WEML). To examine the effect of WEML on cell viability in dermal papillar (DP) cells, 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetra Zolium (MTS) analysis was performed. The changes in the mRNA expression level of the hair loss and hair growth-related genes in dermal papilla cells by WEML treatment were confirmed by quantitative RT-PCR. Results : In dermal papilla (DP) cells, ABTS analysis and MTS analysis of WEML showed antioxidant efficacy and low cytotoxicity. As a result of gene expression analysis through Quantitative RT-PCR, no changes in hair growth-related genes BMP6 and CTNNB1 was confirmed. but inhibitory activity of WEML on hair loss-related genes EGR1, SGK, DKK1, SRD5A1 and SRD5A2 was confirmed. Conclusion : We confirmed that WEML has excellent antioxidant efficacy and a inhibitory activity of hair loss-related genes including 5α-reductase genes. These results suggest that Mangifera indica leaves have a potential activity as a hair loss treatment for hair loss and hair growth. Biochemical or molecular biological research on hair loss is needed.

• YAKHAK HOEJI
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• v.53 no.4
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• pp.206-216
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• 2009

3-Hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (Statins) are potent inhibitors of cholesterol biosynthesis. Cholesterol-lowering therapy using statins significantly reduces the risk of coronary heart disease. Various discovery of statins as bone anabolic agents has spurred a great deal of interest among both basic and clinical bone researchers. In-vitro and some animal studies suggest that statins increase the bone mass by enhancing bone morphogenetic protein-2 (BMP-2)-mediated osteoblast expression. Clinical and animal test results of statins focusing on the prevention and treatment of bone fractures was collected. Three independent literature searches were performed by using from January 1, 2002 to September 2008 for clinical and animal test results. Search term included statins, HMG-CoA reductase inhibitors, pleiotropic effects, fracture, osteoporosis and clinical and animal test. No consensus has been reached whether clinical use of statins has beneficial effects on bone health, partly due to lower statin concentrations because of first-pass metabolism by the liver. Experimental use of statins as stimulators of bone formation suggests that they may have widespread applicability in the field of orthopaedics. With their combined effects on osteoblasts and osteoclasts, statins have the potential to enhance resorption of synthetic materials and improve bone ingrowth. In conclusion, The use of statins in the prevention and treatment of bone fractures requires further study. But observational studies suggest that statins for decreasing bone fractures including osteoporosis have to be considered local direct administration like transdermal or subcutaneous type over oral adminstration.

• Molecules and Cells
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• v.37 no.4
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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.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.33 no.3
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• pp.247-255
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• 2007

The bone morphogenic protein(BMP) can promote migration and growth of mesenchymal cells and initiate process for bone and cartilage formation. Cartilage-derived morphogenic protein(CDMP)-1 and -2 belong to the bone morphogenetic protein family in the transforming growth factor(TGF)-${\beta}$ superfamily. Although pleomorphic adenoma of the salivary glands is an epithelial tumor, it frequently shows ectopic cartilaginous formation with biomolecular studies. The mechanism of pathogenesis in cartilaginous formation is still controversy. We examined the expression and localization of CDMP-1 and -2, in comparison with the localization of cartilaginous matrix proteins, in human normal salivary glands and 20 cases of pleomorphic adenoma using immunohistochemical methods. The results were followed. 1. CMP-1 was immunolocalized in the striated ducts and the intercalated ducts, but not expressed in excretory duct, CDMP-2 was not expressed in the normal salivary glands. 2. CMP-1 was immunolocalized in the ductal cell and cuboidal neoplastic myoepithelial cells around the chondroid areas of the pleomorphic adenomas, whereas these molecules were not localized in the spindle-shaped neoplastic myoepithelial cells of the myxoid element in these tumors. CDMP-2 was expressed neither in normal salivary glands nor in any elements of the pleomorphic adenomas. 3. In transmission electron microscopic view, the tumor cells are composed of modifed myoepithelial cells between hyaline and myxoid stroma. 4. In Immuno-blot analysis, strong overexpression of CDMP-1 was frequently seen in pleomorphic adenomas, but the level of CDMP-2 was expressed minimally in pleomorphic adenoma. From the these results, it should be suggested that undifferentiated neoplastic myoepithelial cells around the chondroid areas expressed CDMP-1 and suggested that this molecule may play a role in the differentiation of neoplastic myoepithelial cells in pleomorphic adenoma, but not CDMP-2.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.5
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• pp.635-647
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• 2014

Unfertilized oocytes age inevitably after ovulation, which limits their fertilizable life span and embryonic development. Rapamycin affects mammalian target of rapamycin (mTOR) expression and cytoskeleton reorganization during oocyte meiotic maturation. The goal of this study was to examine the effects of rapamycin treatment on aged porcine oocytes and their in vitro development. Rapamycin treatment of aged oocytes for 24 h (68 h in vitro maturation [IVM]; $44h+10{\mu}M$ rapamycin/24 h, $47.52{\pm}5.68$) or control oocytes (44 h IVM; $42.14{\pm}4.40$) significantly increased the development rate and total cell number compared with untreated aged oocytes (68 h IVM, $22.04{\pm}5.68$) (p<0.05). Rapamycin treatment of aged IVM oocytes for 24 h also rescued aberrant spindle organization and chromosomal misalignment, blocked the decrease in the level of phosphorylated-p44/42 mitogen-activated protein kinase (MAPK), and increased the mRNA expression of cytoplasmic maturation factor genes (MOS, BMP15, GDF9, and CCNB1) compared with untreated, 24 h-aged IVM oocytes (p<0.05). Furthermore, rapamycin treatment of aged oocytes decreased reactive oxygen species (ROS) activity and DNA fragmentation (p<0.05), and downregulated the mRNA expression of mTOR compared with control or untreated aged oocytes. By contrast, rapamycin treatment of aged oocytes increased mitochondrial localization (p<0.05) and upregulated the mRNA expression of autophagy (BECN1, ATG7, MAP1LC3B, ATG12, GABARAP, and GABARAPL1), anti-apoptosis (BCL2L1 and BIRC5; p<0.05), and development (NANOG and SOX2; p<0.05) genes, but it did not affect the mRNA expression of pro-apoptosis genes (FAS and CASP3) compared with the control. This study demonstrates that rapamycin treatment can rescue the poor developmental capacity of aged porcine oocytes.