• Korean Journal of Ichthyology
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• v.19 no.2
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• pp.107-111
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• 2007

Sex differentiation of the brown croaker Miichthys miiuy (Basilewsky) is described from hatching to the 120th day post-hatching (dph) (water temperature $24^{\circ}C$). Primordial germ cells (PGCs) were observed on the 20th dph (10.4 mm total length (TL), 0.14 g body weight (BW), and began to protrude into the peritoneal cavity from the 40th dph (19.4 mm TL, 0.39 g BW). On the 65th dph (31.3 mm TL, 0.93 g BW, $1,560D^{\circ}$ (degree-days)), initial ovarian differentiation was identified by the PGCs with condensed chromatin, and their transformation into meiotic oocytes. By the 120th dph (4.60 mm TL, 1.38 g BW, $2,880D^{\circ}$), the oocytes were in the perinucleolus stage and had increased from 20 to $40{\mu}m$ in diameter. While ovaries gradually grew after sex was differentiated, testes continued to multiply from the 65th dph. On the 80th dph (37.9 mm TL, 1.39 g BW, $1,920D^{\circ}$), the beginning of testis lobule formation was indicated by the occurrence of spermatogonial cysts enveloped by somatic cells in some of the testes. On the 120th dph, the testis lobules of some of the fish contained all germ cell stages through to the spermatocytes. Therefore, the sex differentiation type of the brown croaker is identified as gonochoristic.

• Korean Journal of Medicinal Crop Science
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• v.2 no.1
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• pp.51-59
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• 1994

This study was carried out to investigate the optimal propagation condition through stem node tissue culture of yam supplemented with concentration levels of growth regulators and activated charcoal in 1 /8MS white media. For shoot induction and plant regeneration from yam stem node tissue wre more effectivein combination of auxin, kinetin and activated charcoal than combination of auxin and kinetin. Shoot induction and plant regeneration were more effective in 1/8 medium suplemented with 1AA $2mg\;/\;\ell,\;kinetin\;2mg\;/\;\ell$ and activated charcoal $1g\;/\;\ell$ than in white medium, but in case of white medium, Shoot induction and plant regeneration wre effective in culture medium supplemented with NAAa $4mg\;/\;\ell,\;kinetin\;2mg\;/\;\ell$ and activated charcoal $1g\;/\;\ell$. Growth status in optimal propagation condition from yam stem node tissue brought the results of multiple plant, elongated plant height and increase in number of leaves and roots, but tissue culture effect by kinds of media in number of multiple plant were more effective in 1 /8MS medium than in white medium.

• Journal of Life Science
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• v.17 no.5 s.85
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• pp.678-686
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• 2007

Human mesenchymal stem cells(hMSC), that have been reported to be present in bone marrow, adipose tissues, dermis, muscles and peripheral blood, have the potential to differentiate along different lineages including those forming bone, cartilage, fat, muscle and neuron. Therefore, hMSC are attractive candidates for cell and gene therapy. The optimal conditions for hMSC expansion require medium supplemented with fetal bovine serum(FBS). Some forms of cell therapy will involve multiple doses, raising a concern over immunological reactions caused by medium-derived FBS proteins. Previously, we have shown that hADSC can be cultured in human serum(HS) during their isolation and expansion, and that they maintain their proliferative capacity and ability for multilineage differentiation and promote engraftment of peripheral blood-derived CD34 cells mobilized from bone marrow in NOD/SCID mice. In this study we determined whether hADSC grown in HS maintain surface markers expression similar with cells grown in FBS during culture expansion and compared gene expression profile by Affymetrix microarray. Flow cytometry analysis showed that HLA-DR, CD117, CD29 and CD44 expression in HS-cultured hADSC during culture expansion were similar with that in FBS-cultured cells. However, the gene expression profile in HS-cultured hADSC was significantly different from that in FBS-cultured cells. Therefore, these data indicated that HS-cultured hADSC should be used in vivo animal study of hADSC transplantation for direct extrapolation of preclinical data into clinical application.

• Korean Journal of Environmental Biology
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• v.41 no.3
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• pp.325-334
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• 2023

The invasive red swamp crayfish, Procambarus clarkii, is native to south-central United States and northeastern Mexico. Recently, it has been being spreading in the wild in South Korea. However, its primary sources, introduction routes, establishment, and expansion in South Korea remain unclear. Here, we analyzed genetic diversity and population genetic structures of its domestic natural populations during early invasion, commercial stock from local aquaria (a suspected introduction source), and original United States population using mitochondrial COI gene sequences for 267 individuals and eight microsatellite markers for 158 individuals. Natural and commercial populations of P. clarkii showed reduced genetic diversity (e.g., haplotype diversity and allelic richness). The highest genetic diversity was observed in one original source population based on both genetic markers. Despite a large number of individuals in commercial aquaria, we detected remarkably low genetic diversity and only three haplotypes among 226 individuals, suggesting an inbred population likely originating from a small founder group. Additionally, the low genetic diversity in the natural population indicates a small effective population size during early establishment of P. clarkii in South Korea. Interestingly, genetic differentiation between natural populations and the United States population was lower than that between natural populations and aquarium populations. This suggests that various genetic types from the United States likely have entered different domestic aquariums, leading to distinct natural populations through separate pathways. Results of our study will provide an insight on the level of genetic divergence and population differentiation during the initial stage of invasion of non-indigenous species into new environments.

• Journal of Korean Society of Forest Science
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• v.75 no.1
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• pp.25-31
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• 1986

In order to contribute to the Korean tea-plant culture and tea industry by means of increasing the production of tea-plants, I have performed the tissue culture of the organs of the anther, leaf and stem. As for the culture-material, I have used the anther of tea (Thea sinensis) at the tetrad uninucleate microspore stage and used medium of modified Murashige and Skoog as the basal medium supplemented with the growth regulators of NAA and 2, 4-D, yeast, kinetin and others at various concentrations. As for the handling of material, I have followed the common methods of sterilization and microtoming and paraffine imbedding method and observed systematically periodic changes of the microspores in culture. I have divided the leaf, stem and root into segments and sterilized them and used the modified Murashige and Skoog as the basal medium and observed the differentiation of roots and callus and the results are as follows. 1. In case of anther, I have found 2n callus was found in 30 out of 100 segments in M2 medium. 2. The differentiation of roots appeared in 24.5% of total leaf segments cultured and in 50.5% of stem and in 43.9% of root. 3. When the differentiation of stem in different parts was observed, the most frequent differentiation was found in the second part of all the 4 parts. 4. The most frequent formation of callus was noticed from the anther-walls in case of anther culture and from the veins in case of leaf culture. It is concluded that the seedlings of tea-plant could be multiplied most by means of tissue culture of the second part of the tea-plant stem and reduction in the expenditures of tea-plant propagation was possible through tissue culture.

• The korean journal of orthodontics
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• v.27 no.2
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• pp.335-347
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• 1997

Vitamin D is known to exert its action by activating DNA and RBA within target cells to produce proteins and enzymes that can be used in bone resorption process. Particularly, the active form of vitmain D, 1,25-dihydroxycholecalciferol $[1,25-(OH)_2D_3]$, is considered to be one of the most potent stimulators of osteoclatic acitivity in vitro. The purpose of this study was to evaluate the effect of 1,25-Dihydroxyvitamin $D_3$ on the avtivity of periodotal ligament cells and, the experimental tooth movement. Human periodontal ligament cells were collected from the first premolar tooth extracted for the orthodontic treatment, and were incubated in the environment of $37^{\circ}C$, 5% $CO_2$ and 95% humidity. Microtitration(MIT) assay was done at 10, 25, 50 and 100ng/ml of 1,25-Dihydroxyvitamin $D_3$. 21 Sprague-Daft rats were divided into a control gmup(3), and experimental groups(18) where 100g of force from helical spring was applied across the maxillary incisors 1,25-Dihydroxyvitamin $D_3$ was injected into periodontal ligament at the mesial or distal surface of maxillary incisors so that we can compare the control side and the experimental side. Expreimental groups were sac rifled at 12, 24, 36, 48, 72hours and 7 days after force application, respectively. And the obtained tissues were evaluated histologically. The observed results were as follows. 1. The activity of periodontal ligament cells in l0ng/ml or 25ng/ml of 1,25-Dihydroxyvitamin $D_3$ 1,25-Dihydroxyvitamin $D_3$ was not significantly different to the control at the cultivation of 1, 2 and 3 days. 2. The activity of periodontal ligament cells was significantly increased at 3 days in 50 ng/ml of 1,25-Dihydroxyvitamin $D_3$ and 2, 3 days in 100g/ml of 1,25-Dihydroxyvitamin $D_3$. 3. Up to 7 days after force application, there was no difference in osteoblastic activity, tearing of periodontal ligament and proliferation of capillary at tension side between 1,25-Dihydroxyvitamin $D_3$ injection side and the control side. 4. The osteoclastic activity and the resorption of alveolar bone was greater in 1,25-Dihydroxyvitamin $D_3$ injection side than the control side at 36 hours after force application.

• Tuberculosis and Respiratory Diseases
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• v.59 no.2
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• pp.142-150
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• 2005

Background : Continuous growth stimulation by various factors, as well as chronic oxidative stress, may co-exist in many solid tumors, such as lung cancer. A new family of antioxidant proteins, the peroxiredoxins (Prxs), have been implicated in the regulation of many cellular processes, including cell proliferation, differentiation and apoptosis. However, a real pathophysiological significance of Prx proteins, especially in lung disease, has not been sufficiently defined. Therefore, this study was conducted to investigate the distribution and expression of various Prx isoforms in lung cancer and other pulmonary conditions. Method : Patients diagnosed with lung cancer, and who underwent surgery at the Ajou Medical Center, were enrolled. The expressions of Prxs, Thioredoxin (Trx) and Thioredoxin reductase (TR) were analyzed using proteomic techniques and the subcellular localization of Prx proteins was studied using immunohistochemistry on normal mouse lung tissue. Result : Immunohistochemical staining has shown the isoforms of Prx I, II, III and V are predominantly expressed in bronchial and alveolar lining epithelia, as well as in the alveolar macrophages of the normal mouse lung. The isoforms of Prx I and III, and thioredoxin were also found to be over-expressed in the lung cancer tissues compared to their paired normal lung controls. There was also an increased amount of the oxidized form of Prx I, as well as a putative truncated form of Prx III, in the lung cancer samples when analyzed using 2-dimensional electrophoresis. In addition, a 43 kDa intermediate molecular weight protein band, and other high molecular weight bands of over 20 kDa, recognized by the anti-Prx I antibody, were present in the tissue extracts of lung cancer patients on 1-Dimensional electrophoresis, which require further investigation. Conclusion : The over-expressions of Prx I and III, and Trx in human lung cancer tissue, as well as their possible chaperoning function, may represent an attempt by tumor cells to adjust to their microenvironment in a manner advantageous to their survival and proliferation, while maintaining their malignant potential.

• Tuberculosis and Respiratory Diseases
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• v.52 no.6
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• pp.608-615
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• 2002

Background : ADA is an enzyme found in most cells, and is involved in purine metabolism, but its chief role concerns the proliferation and differentiation of lymphocytes, especially T-lymphocytes. For that reason ADA has been looked on as a marker of cell-mediate immunity, which is the key mechanism of the tuberculous pleural effusion. Thus, the pleural fluid ADA activity is increased in the tuberculous pleural effusion. Age associated immune decline is characterized by decreases in both B and T-lymphocyte function and the former may be largely a result of the latter. Therefore, the pleural fluid ADA activity would be lower in old rather than in young, patients with tuberculous pleural effusion. We studied the relationship between age, and pleural fluid ADA activity, in patients with tuberculous pleural effusion. Materials and Methods : In the 46 patients with tuberculous pleural effusion enroll in this study, the pleural fluid ADA activities were measured by means of an automated kinetic method. Results : The mean age of the patients was $53.0{\pm}22.0$ years, with a male to female ratio of 30:16. The patients were divided into two groups, young patients, regarded as < 65 and old regarded as ${\geq}65$ years with 28 and 18 patients, respectively. The pleural fluid ADA activity in both groups show significant differences : $99.4{\pm}22.6$ IU/L(young patients) Vs. $75.8{\pm}30.9$ IU/L(old patients)(p<0.05), but a negative correlation with age (r=-0.311, p<0.05). Conclusion : Although pleural fluid ADA activity was not adequately increased, tuberculous pleural effusion, in older patients, would have to be considered clinically suspicious tuberculous pleural effusion.

• 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.

• Journal of Periodontal and Implant Science
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• v.35 no.1
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• pp.199-216
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• 2005

치주치료의 가장 중요한 목적은 상실된 치주조직의 형태적, 기능적 재건이다. 법랑기칠 단백질 유도체(enamel matrix derivative: EMD)는 치주 병소에 사용시 상피세포의 증식을 억제하며 치주인대 및 백악아세포를 활성화시켜 무세포성 백악질 및 치주인대와 골조직의 생성을 유도한다고 보고되고 있다. 또한 법랑기질 단백칠 유도체는 골모세포의 증식 및 분화를 촉진시키며 alkaline phosphatase의 활성 및 mineralized nodule의 형성을 촉진시킨다고 보고되고 있다. 이에 본 연구에서는 토끼 두개골 결손부에 법랑기질 단백질 유도체와 이종골 이식재를 이식한 후 골밀도를 방사선학적으로 분석하고, 신생골 형성 및 주변 조직 반응을 조직학적으로 관찰, 평가하고자 하였다. 토끼 두개골에 6mm trephine bur(외경 8mm)를 이용하여 경뇌막에 손상을 주지 않도록 하면서 4개의 결손부를 형성하였다. 아무것도 이식하지 않은 군을 음성 대조군으로, 이종골 이식재 ($Bio-Oss^{(R)}$, Geistlich, Wolhusen, Switzerland)을 이식한 군을 양성 대조군으로 설정하였다. 법랑기질 단백질 유도체 ($Emdogain^{(R)}$, Biora, Inc., Sweden)만 이식한 군과 법랑기질 단백질 유도체와 이종골 이식재를 혼합하여 이식한 군을 설험군으로 설정하였다. 각각의 재료를 이식한 후 비흡수성 차폐막 ($Tefgen^{(R)}$, Lifecore Biomedical, Inc., U.S.A.)을 위치시키고 흡수성 봉합사로 일차봉합을 시행하였다. 각 군당 술 후 1, 2, 4주의 치유기간을 설정하였다. 동물을 희생시킨 후 두개골을 절제하여 먼저 방사선학적인 골밀도측정을 시행한 후 10% formalin에 고정한 후 통법에 따라 조직표본을 제작하여 광학현미경으로 관찰하였다. 1. 방사선학적인 평가에서 1, 2, 4주에 대조군과 법랑기질 단백질 유도체만 이식한 군과 비교해 이종골 이식재만 이식한 군과 이종골 이식재에 법랑기질 단백질 유도체를 이식한 군에서 더 큰 골의 밀도를 보이고 있었다 (P<0.01). 하지만, 동일한 시기에 대조군과 법랑기질 단백질 유도체만 이식한 군과의 차이는 발견할 수 없었으며 (P>0.05), 이종골 이식재만 이식한 군과 이종골 이식재에 법랑기질 단백질 유도체를 이식한 군의 차이 또한 발견할 수 없었다 (P>0.05). 2. 조직학적인 평가에서 1, 2, 4주에 대조군과 법랑기질 단백질 유도체만 이식한 군과 비교해 이종골 이식재만 이식한 군과 이종골 이식재에 법랑기질 단백질 유도체를 이식한군에서 골의 형성이 더 진행됨을 알 수 있었다. 법랑기질 단백질 유도체만 이식한 군이 대조군보다 2주에서 더 많은 신생골을 볼 수 있었으며, 이종골 이식재에 법랑기질 단백질 유도체를 이식한 군이 이종골 이식재만 이식한 군보다 1, 2주에서 더 많은 신생골을 관찰할 수 있었다. 이상의 결과에서 법랑기질 단백질 유도체는 토끼 두개골 결손부 치유단계에서 초기 골 형성을 촉진하는 것으로 사료되며 골 이식시에 법랑기질 단백질 유도체를 적용하는 것은 유용한 술식으로 사료된다.