• 한국동물생명공학회지
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• 제34권1호
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• pp.2-9
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• 2019

Because mesenchymal stem cells (MSCs) maintain distinct capacities with respect to self-renewal, differentiation ability and immunomodulatory function, they have been highly considered as the therapeutic agents for cell-based clinical application. Of particular, differentiation condition alters characteristics of MSCs, including cellular morphology, expression of gene/protein and cell surface molecule, immunological property and apoptosis. However, the previous results for differentiation-related apoptosis in MSCs have still remained controversial due to varied outcomes. Therefore, the present study aimed to disclose periodical alterations of pro- and anti-apoptosis in MSCs under differentiation inductions. The human dental pulp-derived MSCs (DP-MSCs) were differentiated into adipocytes and osteoblasts during early (1 week), middle (2 weeks) and late (3 weeks) stages, and were investigated on their apoptosis-related changes by Annexin V assay, qRT-PCR and western blotting. The ratio of apoptotic cell population was significantly (p < 0.05) elevated during the early to middle stages of differentiations but recovered up to the similar level of undifferentiated state at the late stage of differentiation. In the expression of mRNA and protein, whereas expressions of pro-apoptosis-related makers (BAX and BAK) were not altered in any kind and duration of differentiation inductions, anti-apoptosis marker (BCL2) was significantly (p < 0.05) elevated even at the early stage of differentiations. The recovery of apoptotic cell population at the late stage of differentiation is expected to be associated with the response by elevation of anti-apoptotic molecules. The present study may contribute on understanding for cellular mechanism in differentiation of MSCs and provide background data in clinical application of MSCs in the animal biotechnology to develop effective and safe therapeutic strategy.

• 아동학회지
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• 제38권1호
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• pp.233-249
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• 2017

Objective: This study examined the effects of maternal grandmothers' positive parenting behavior and mothers' self-differentiation and positive parenting behavior on self-concept in late childhood. Methods: A total of 341 $5^{th}$- and $6^{th}$- grade elementary school children and their mothers participated in the study. Children were surveyed regarding perceptions of their mothers' positive parenting behavior and their own self-concept. Mothers responded to questionnaires regarding both positive parenting behavior of their mothers and their own self-differentiation. The data were analyzed using SEM. Results: Maternal grandmothers' positive parenting behavior had an indirect effect through mothers' self-differentiation and positive parenting behavior on children's self-concept. Mothers who perceived positive parenting behavior in their mothers in their childhood showed positive parenting behavior (as perceived by their children), and those children reported a higher level of self-concept. Mothers who perceived their mother's parenting behavior in their childhood more positively reported a higher level of self-differentiation, and those mothers showed more positive parenting behavior (as perceived by their children), which led to more positive self-concept in children. Conclusion: The findings specify the pathways from maternal grandmothers' positive parenting behavior to children's self-concept through mothers' self-differentiation and positive parenting behavior in an aspect of the intergenerational transmission of parenting. The findings also emphasize the importance of mothers' positive parenting behavior for positive self-concept in late childhood.

• Journal of Periodontal and Implant Science
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• 제36권3호
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• pp.783-796
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• 2006

The periodontium is a topographically complex organ consisting of epithelial tissue, soft and mineralized tissues. Structures comprising the periodontium include the gingiva, periodontal ligament (PDL) , cementum and the alveolar bone. The molecular mechanism of differentiation in PDL fibroblast cells remain unclear. Amino acid transporters play an important role in supplying nutrition to normal and cancer cells and for cell proliferation. Amino acid transport system L is a major nutrient transport system responsible for the Na+-independent transport of neutral amino acids including several essential amino acids. The system L is divided into two major subgroups, the L-type amino acid transporter 1 (LAT1) and the L-type amino acid transporter 2 (LAT2). In this study, the expression pattern of amino acid transport system L was, therefore, investigated in the differentiation of PDL fibroblast cells. To determine the expression level of amino acid transport system L participating in intracellular transport of amino acids in the differentiation of PDL fibroblast cells, it was examined by RT-PCR, observation of cell morphology, Alizaline red-S staining and uptake analysis after inducing experimental differentiation in PDL fibroblast cells isolated from mouse molar teeth. The results are as follows. 1. The LAT1 mRNA was expressed in the early stage of PDL fibroblast cell differentiation. This expression level was gradually reduced by differentiation- inducing time and it was not observed after the late stage. 2. The expression level of LAT2 mRNA was increased in time-dependent manner during differentiation induction of PDL fibroblast cells. 3. There was no changes in. the expression level of 4F2hc mRNA, the cofactor of LAT1 and LAT2, during differentiation of PDL fibroblast cells. 4. The expression level of ALP mRNA was gradually increased and the expression level of Col I mRNA was decreased during differentiation of PDL fibroblast cells. 5. The L-leucine transport was reduced by time from the early stage to the late stage in PDL fibroblast cell differentiation. As the results, it is considered that among neutral ammo acid transport system L in differentiation of PDL fibroblast cells, the LATl has a key role in cell proliferation in the early stage of cell differentiation and the LAT2 has an important role in the late stage of cell differentiation for providing cells with neutral amino acids including several essential amino acids.

• Reproductive and Developmental Biology
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• 제30권2호
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• pp.107-113
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• 2006

Development of placenta is a complex process that is critical for the pregnancy and controlled by many factors including cytokines, hormones, growth factors and apoptotic molecules. Recently, it has been shown that progranulin (PGRN) functions in growth of embryo and trophectoderm as well as cell migration. To initiate understanding the role of PGRN in human placental development, we investigated the expression of PGRN mRNA and protein in early and late gestation human placentas, term cytotrophoblast cells and two choriocarcinoma cell lines, JEG-3 and Jar. Reverse transcriptase polymerase chain reaction identified mRNAs derived from the PGRN gene in all samples. Immunoblot analysis showed that PGRN proteins are present in early and late gestation human placentas with decreasing levels over gestation and that PGRN proteins are present in normal and transformed trophoblast cells. Immunohistochemical analysis using paraformaldehyde-fixed tissue sections taken from early and late stages of pregnancy showed that PGRN proteins are present in cytotrophoblast cells, syncytiotrophoblast and extravillous cytotrophoblast cells and that expression pattern of PGRN differed according to the stage of cell differentiation. The results of this study are consistent with the hypothesis that PGRN proteins have critical roles in placental development and suggest that PGRN may function in trophoblast cell growth and differentiation.

• 약학회지
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• 제38권3호
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• pp.238-249
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• 1994

The purpose of this research was to investigate effects of baicalin on the differentiation of preadipocytes, 3T3-L1, and to characterize the action of baicalin that affect the responses of 3T3-L1 cells during differentiation. In various culture conditions, effects of baicalin and adrenoreceptor agonists such as phenylephrine(PE) and isoproterenol(IPR) on cell differentiation were examined. Also, effects of the drugs on differentiation, triglyceride(TG) contents, expression of insulin receptor, cAMP contents, the cytosolic $Ca^{2+}$ levels, and amount of calmodulin(CaM) were examined. The results suggest that baicalin has adrenergic receptor blocking activity during the process of differentiation of 3T3-L1 cells and that in the early stage of the adipose conversion, the effect of baicalin on the adipocyte differentiation is mediated by the regulation of insulin receptor expression, but by alterations of the cAMP and the calcium metabolism in the late stage. These results also suggest that the action of baicalin may be significant in the lipid metabolism, lipogenic and lipolytic pathways, of adipose cells.

• International Journal of Oral Biology
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• 제33권1호
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• pp.33-43
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• 2008

In the process of bone remodeling, mineral phase of bone is dissolved by osteoclasts, resulting in elevation of calcium concentration in micro-environment. This study was performed to explore the effect of high extracellular calcium ($Ca{^{2+}}_e$) on mineralized nodule formation and on the expression of progressive ankylosis (Ank), plasma cell membrane glycoprotein-1 (PC-1) and osteopontin by primary cultured mouse calvarial cells. Osteoblastic differentiation and mineralized nodule formation was induced by culture of mouse calvarial cells in osteoblast differentiation medium containing ascorbic acid and ${\beta}$-glycerophosphate. Although Ank, PC-1 and osteopontin are well known inhibitors of mineralization, expression of these genes were induced at the later stage of osteoblast differentiation during when expression of osteocalcin, a late marker gene of osteoblast differentiation, was induced and mineralization was actively progressing. High $Ca{^{2+}}_e$(10 mM) treatment highly enhanced mRNA expression of Ank, PC-1 and osteopontin in the late stage of osteoblast differentiation but not in the early stage. Inhibition of p44/42 MAPK activation but not that of protein kinase C suppressed high $Ca{^{2+}}_{e^-}$induced expression of Ank, PC-1 and osteopontin. When high $Ca{^{2+}}_e$(5 mM or 10 mM) was present in culture medium during when mineral deposition was actively progressing, matrix calcifiation was significantly increased by high $Ca{^{2+}}_e$. This stimulatory effect was abolished by pyrophosphate (5 mM) or levamisole (0.1-0.5 mM), an alkaline phosphatase inhibitor. In addition, probenecid (2mM), an inhibitor of Ank, suppressed matrix calcification in both control and high $Ca{^{2+}}_{e^-}$treated group, suggesting the possible role of Ank in matrix calcification by osteoblasts. Taken together, these results showed that high $Ca{^{2+}}_e$ stimulates expression of Ank, PC-1 and osteopontin as well as matrix calcification in late differentiation stage of osteoblasts and that p44/42 MAPK activation is involved in high $Ca{^{2+}}_{e^-}$induced expression of Ank, PC-1 and osteopontin.

• Journal of Animal Science and Technology
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• 제53권3호
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• pp.223-226
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• 2011

The current study was undertaken to determine the effect of retinoic acid (RA) on proliferation and differentiation of preadipocytes from male and female pigs. The preadipocytes were isolated from new-born male and female pigs by collagenase digestion and washed three times one day after seeding (designated as day 0 of culture). RA was included in the media at various concentratives from day 0 to 2. The cell number was measured on day 2 with hematocytometer after trypsin digestion. Cell differentiation was determined on day 6 by measuring glycerol-3-phosphate dehydrogenase activity. RA (0.1, 1 and 10 uM) showed no effect on proliferation of preadipocytes from both male and female pigs. However, RA significantly decreased differentiation of pig preadipocytes. Degree of differentiation with 0.1 uM, 1 uM and 10 uM of RA treatment was 80%, 41% and 29% respectively, compared with control. Similar inhibitory effect was found in the female pigs; 77%, 28% and 16% respectively. It is interesting that RA treated on cell proliferation stage had no effect on proliferation but had a strong inhibitory effect on differentiation which is happening in the late stage of cell culture.

• 한방비만학회지
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• 제15권1호
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• pp.38-44
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• 2015

Objective: Skeletal muscle is a crucial tissue from the perspectives of mitochondrial dysfunction and insulin resistance, it is formed by myogenesis which is dynamic multistep process to be myotubes. The authors could found that root of Atractylodes macrocephala Koidzumi (Atractylodis Rhizoma Alba, ARA) enhanced glucose and lipid metabolism in C2C12 myotubes via mitochondrial regulation. However its action in myogenesis process is not known. The aim of this work was the study of ARA on proliferation, differentiation and hypertrophy in C2C12 cells. Methods: To study proliferation phase, cells were incubated in growth medium with or without ARA (0.2 or 1.0 mg/ml) for 24 hours. To examine differentiation, at 70% confluence, cells were transferred in differentiation medium both with/without ARA (0.2 or 1.0 mg/ml) for 96 hours. And after 72 hours of differentiation, cells were treated with or without ARA (0.2 or 1.0 mg/ml) for 24 hours, the genesis of hypertrophy in myotubes were analyzed. Results: In proliferation phase, ARA could make difference in morphologic examination. In differentiation phase, it also made morphologic difference furthermore ARA (1.0 mg/ml) increased mRNA expressions of Myogenic regulatory factors and muscle-specific proteins synthesis. In late differentiation, ARA induced hypertrophic morphological changes in neo-formed myotubes. Conclusions: ARA might control cell cycle promoting myogenesis and hypertrophy in C2C12 cells.

• 한국광물학회지
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• 제26권4호
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• pp.299-312
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• 2013

홍천 카보나타이트-포스코라이트 복합체를 형성하는 주 구성광물인 자철석은 각각 세 시기에 걸쳐 정출되었으며 후기로 가면서 함량이 점차 감소한다. 자철석에 대한 전자현미분석결과 Ti, V은 미량 검출되지만 초기에서 후기로 가면서 증가하는 경향을 보여준다. 반면, Mg, Mn은 뚜렷이 감소하는데 이는 일반적인 카보나타이트질 마그마 분화특성을 잘 나타낸다. Al 또한 카보나타이트와 포스코라이트에서 감소하는 경향을 보여주며, Cr은 대부분 검출한계 미만을 나타내나 후기 포스코라이트에 와서는 미량 정출된다. 자철석은 초기에는 $Fe^{2+}$가 주로 $Mg{2+}$와 $Mn^{2+}$에 의해 치환되고, $Fe^{3+}$는 $Al^{3+}$에 의한 치환되는 양상이 주를 이루었으나 후기에 와서는 감소하면서 거의 순수한 자철석 조성을 갖게 된다. V의 증가와 Mn의 감소는 마그마 분화가 산소분압이 점차 감소하는 환경에서 진행되었음 나타내고, 감람석, 금운모의 부재와 더불어 자철석의 Mg, Al, Cr 및 Ti 원소들의 함량이 낮은 것은 홍천 카보나타이트-포스코라이트 복합체가 결핍된 모마그마로부터 생성되었음을 지시한다. 특히, 후기로 가면서 철질 탄산염광물과 석영의 산출이 두드러지면서 전형적인 카보나타이트-포스코라이트 복합체에 비해 연구지역 자철석의 Mg 함량이 적게 산출되는 것은 마그마 분화가 최후기까지 진행되었음을 시사한다.

• Applied Microscopy
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• 제22권2호
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• pp.97-117
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• 1992

In order to study process of spermiogenesis of the Korean greater horseshoe bat, Rhinolophus ferrumequinum korai, the cycle of seminiferous epithelium was examined by the light and electron microscope and the following results were obtained based on the epithelial cell differentiation. 1. Spermiogenesis occurred from early July to mid-Octber, and spermatogenic activity was vigorous from mid-August to late September. Spermatocytes including spermatogonia were found to be degenerated in only July. It is deduced that the degeneration serves as the mechanism to regulate effective use of energy to prepare for mating and hibernating periods, and regulation of breeding cycle. 2. Spermiogenesis of the Korean greater horseshoe bat was divided according to differentiation of the cell structure, into Golgi, cap, acrosome, maturation and spermiation phases; Golgi, cap and spermiation phases were further divided into two steps of early and late phase respectively, and acrosome phase into three steps of early, mid and late phases, and maturation phase has only one step. Hence, the spermiogenesis consists of ten phases. The first research was done in this article on the changes of chromatin with nucleus, the time of appearance and disappearance of chromatin granules, in case of Korean greater horseshoe bat (Rhinolophus ferrumequinum korai). Chromatin granule began to be condensed in late Golgi and the condensation proceeded to form an irregular mass of a electron-dense chromatin in a form of circular cylinder in the center of nucleus at the phase of maturation. Finally, the chromatin condensation proceeded and perfect nucleus of sperm with homogeneous density was formed when the sperm was separated from Sertoli cell. Therefore, appearance and disappearance of chromatin granules occurred in the period of time between late Golgi and maturation phase, The tail of sperm began to develop in early cap phase, Numerous lipid droplets were obseved in the cytoplasm of spermatids during the maturation phase, which seemed to be used as energy source necessary to make mature sperm during spermiogenesis.