• Title/Summary/Keyword: 분화론

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Establishment of Embryonic Stem Cell Line from Isolated Blastomeres from Mouse Preimplantation Embryos (생쥐 초기 배아에서 분리한 할구를 이용한 배아줄기세포주 확립)

  • Lim, Chun Kyu;Sung, Ji Hye;Choi, Hye Won;Cho, Jae Won;Shin, Mi Ra;Jun, Jin Hyun
    • Clinical and Experimental Reproductive Medicine
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    • v.33 no.1
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    • pp.25-33
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    • 2006
  • Objective: The aim of this study was to investigate whether embryonic stem (ES) cells can be established from isolated blastomeres of mouse embryos. Methods: Blastomeres were separated from mouse (C57Bl/6J) 2- or 4-cell embryos. Isolated blastomeres or whole 4-cell embryos were co-cultured with mitosis-arrested STO feeder cells in DMEM supplemented with recombinant murine leukemia inhibitory factor and ES-qualified fetal bovine serum. After the tentative ES cell lines were maintained from isolated blastomeres or whole embryos, some of them were frozen and the others were sub-cultured continually. Characteristics of tentative ES cell lines as were evaluated for specific genes expressions with immunocytochemistry and RT-PCR. Results: One ES cell line (3.0%) was established from isolated blastomere of 2-cell embryo and one cell line (4.0%) from isolated two blastomeres of 4-cell embryo. And five cell lines (16.7%) were established from whole 4-cell embryos. Both cell lines from isolated blastomere and whole embryo expressed mouse ES cell specific markers such as SSEA-1, Oct-4 and alkaline phosphatase. Marker genes of three germ layers were expressed from embryoid bodies of both cell lines. Conclusion: This study suggests that mouse ES cells could be established from isolated blastomeres, although the efficiency is lower than whole embryos. This animal model could be applied to establishment of autologous human ES cells from biopsied blastomeres of preimplantation embryos in human IVF-ET program.

Correlation of Proliferating Cell Nuclear Antigen (PCNA) Expression and S-phase Fraction, Survival Rate in Primary Non-Small Cell Lung Cancer (원발성 비소세포 폐암에서 PCNA의 발현정도와 암세포의 분열능 및 생존률과의 관계)

  • Yang, Sei-Hoon;Kim, Hak-Ryul;Gu, Ki-Seon;Jung, Byung-Hak;Jeong, Eun-Taik
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.4
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    • pp.756-765
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    • 1997
  • Background : To study the prognosis of patients with lung cancer, many investigators have reported the methods to detect cell proliferation in tissues including PCNA, thymidine autoradiography, flow cytometry and Ki-67. PCNA, also known as cyclin, is a cell related nuclear protein with 36KD intranuclear polypeptide that is maximally elevated in S phase of proliferating cells. In this study, PCNA was identified by paraffin-embedding tissue using immunohistochemistry which has an advantage of simplicity and maintenance of tissue architecture. The variation of PCNA expression is known to be related with proliferating fraction, histologic type, anatomic(TNM) stage, degree of cell differentiation, S-phase fraction and survival rate. We analyzed the correlation between PCNA expression and S-phase fraction, survival. Method : To investigate expression of PCNA in primary lung cancer, we used immunohistochemical stain to paraffin-embedded sections of 57 resected primary non-small cell lung cancer specimen and the results were analyzed according to the cell type, cell differentiation, TNM stage, S-phase fraction and survival. Results : PCNA expression was divided into five group according to degree of staging(-, +, ++, +++, ++++). Squamous cell type showed high positivity than in adenocarcinoma. Nonsignificant difference related to TNM stage was noticed. Nonsignificant difference related to degree of cell differentiation was noticed. S-phase fraction was increased with advance of PCNA positivity, but it could not reach the statistic significance. The 2 year survival rate and median survival time were -50% 13 months, +75% 41.3 months, ++73% 33.6 months, +++67% 29.0 months, ++++25% 9 months with statistic significance (P<0.05, Kaplan-Meier, generalized Wilcox). Conclusion : From this study, PCNA expression was high positive in squamous cell cancer. And, there was no relationship between PCNA positivity and TNM stage, cellular differentiation or S-phase fraction. But, the patients with high positive PCNA staining showed poor survival rate than the patients with lower positive PCNA staining (p<0.05). It was concluded that PCNA immunostaining is a simple and useful method for survival prediction in paraffin embedded tissue of non-small cell lung cancer.

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The Effects of Echinacea Extract on the Gene Expression of Monocytes and Monocyte-derived Dendritic Cells (Echinacea 추출물이 단구와 단구유래 수지상세포의 유전자발현에 미치는 효과)

  • Park, Jun Eun;Choi, Kang Duk;Kim, Sung Hwan;Hahm, Dae-Hyun;Seo, Jong Jin
    • Clinical and Experimental Pediatrics
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    • v.48 no.7
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    • pp.779-788
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    • 2005
  • Purpose : Echinacea, a traditional plant medicine has been used as immune-stimulant. Recent studies have revealed that extract of Echinacea has immunostimulatory effects on human blood mononuclear cells. This study was designed for the purpose of screening the genes associated with immunologic effects of Echinacea on monocytes and dendritic cells using a cDNA microarray chip. Methods : $CD14^+$ monocyte cells were cultured for one day with Echinacea extract(final concentration : $50{\mu}g/mL$) in experiment 1, but were cultured without Echinacea in experiment 2. The gene expression of these cultured monocytes was analyzed using the cDNA microarray chip. Dendritic cells produced from $CD14^+$ monocyte were cultured for five days with GM-CSF and IL-4, and then cultured for one day with Echinacea in experiment 3, but were done without Echinacea in experiment 4. Results : In experiments 1 and 2, there were 17 significantly expressed genes with average expression ratios above 2.5, including interferon gamma-inducible protein 30(IFI 30), CDC(cell-division-cylcle)-like kinase 2(CLK 2), syndecan binding protein(syntenin), superoxide dismutase 2, etc. In experiments 3 and 4, there were 24 gene, with significantly expressed genes were 24 genes, which were insulin-like growth factor 2(somatomedin A), methyl-CpG binding domain protein 3, IFI 30, small inducible cytokine subfamily A, member 22, etc. The genes encoding CD44, IFI 30, mannose receptor C type 1(MRC 1), chemokine receptor 7(CCR 7), CLK 2, syntenin and cytochrome C oxidase subunit VIII were significantly expressed in both monocytes and dendritic cells cultured with Echinacea. Conclusion : This study employed a cDNA microarray chip to elicit the immune-associated gene profile; the expression was enhanced by Echinacea in CD14+ monocytes and dendritic cells. Thus we laid the basis for the quantitative and functional analysis of genes induced by Echinacea in monocytes and monocyte-derived dendritic cells.

Very Small Putative Stem Cells Detected in Human Endometrium (인간 자궁내막에서 발견되는 극소형 추정줄기세포)

  • Choi, Jong-Ryeol;Joo, Jong-Kil;Jun, Eun-Sook;Ko, Kyoung-Rae;Lee, Hong-Gu;Lee, Kyu-Sup;Kim, Won-Gyu
    • Clinical and Experimental Reproductive Medicine
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    • v.37 no.2
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    • pp.99-113
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    • 2010
  • Objective: It has been recently reported that very small stem cells with pluripotency are detected in murine and human. The purposes of this study are to confirm whether very small putative stem cells (VSPSCs), which have the proper characteristics of stem cells as well as the expression of stem cell markers, are detected in human endometrium. Methods: The endometrial cells of 5 women, which were obtained by endometrial biopsy, were cultured for 2 weeks and were confirmed for the expressions of alkaline phosphatase, OCT-4 and CXCR4 by immunochemistry. Subsequently VSPSCs were separated by percoll density gradient method and were cultured. Also VSPSCs and their derived cells were confirmed for the expressions of OCT-4 and CXCR4. Results: The colonies, which is composed with VSPSCs less than 3 ${\mu}m$ and the 5~15 ${\mu}m$ sized hyperchromatic round cells, were detected in the endometrium of all of women and showed the strong expressions of alkaline phosphatase, OCT-4 and CXCR4. In culture after the separation of VSPSCs by percoll, these cells showed the morphological and functional characteristics of stem cells; self-renewal, colony formation, embryoid body-like formation and differential plasticity. VSPSCs formed gradually the 5~15 ${\mu}m$ sized hyperchromatic round cells and the 10~20 ${\mu}m$ sized sphere-shaped cells by cell-to-cell aggregation or cell fusion. Then these cells differentiated the various cells including fibroblast-like cells, nerve-like cells and endothelium-like cells. VSPSCs and their derived cells often showed the strong expressions of OCT-4 and CXCR4. Conclusion: VSPSCs less than 3 ${\mu}m$ and their derived cells are detected in human endometrium and these cells have the proper characteristics of stem cells and the expressions of stem cell markers as alkaline phosphatase, OCT-4 and CXCR4.

The Results of Postoperative Radiotherapy for Endometrial Carcinoma (자궁 내막암의 수술 후 방사선치료 결과)

  • Noh, O-Kyu;Lee, Sang-Wook;Ahn, Seung-Do;Choi, Eun-Kyung;Kim, Jong-Hyeok;Kim, Yong-Man;Nam, Joo-Hyun;Mok, Jung-Eun;Kim, Jong-Hoon;Shin, Seong-Soo
    • Radiation Oncology Journal
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    • v.25 no.2
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    • pp.93-100
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    • 2007
  • [ $\underline{Purpose}$ ]: To evaluate the outcome and prognostic factors of postoperative radiotherapy in endometrial carcinoma. $\underline{Materials\;and\;Methods}$: From September 1991 to August 2003, 76 patients with endometrial carcinoma received postoperative adjuvant radiotherapy after hysterectomy at Asan Medical Center. Stage was classified as FIGO I in 41 (53.9%), II in 12 (53.9%), and III in 23 (30.3%). Histologic grade 1, 2 and 3 were in 29 (38.2%), 20 (26.3%), and 27 (35.5%) respectively. Forty two patients received both external beam radiation therapy (EBRT) and intracavitary radiation (ICR), 34 patients were treated with EBRT or ICR alone. EBRT dose was 50.4 Gy, ICR was performed in $4{\sim}6$ fractions with $4{\sim}5\;Gy$ per fraction. Median follow-up period was 51 (range $5{\sim}121$) months. $\underline{Results}$: Five-year overall survival was 89.6%. In univariate analysis, statistically significant factors to overall survival were FIGO stage, lymph node metastasis and histologic grade. In disease free survival, FIGO stage, lymph node metastasis and lymphovascular invasion were significant prognostic factors. Recurrence was seen in 11 patients. Of these, systemic failure was in 10 patients. There were no moderate to severe complications after radiation therapy. $\underline{Conclusion}$: The outcome of postoperative adjuvant radiotherapy in endometrial carcinoma was good. Main pattern of failure after postoperative radiotherapy was distant metastasis. So, adjuvant chemotherapy may help in improving outcome. Further study on chemotherapy in combined with postoperative radiotherapy will be needed, especially for patients with high risk factors such as high FIGO stage, lymphovascular invasion, and high histologic grade.

Expression of EGFR in Non-small Cell Lung Cancer and its Effects on Survival (비소세포 폐암에서 EGFR의 발현률과 생존률에 미치는 영향)

  • Kim, Hak-Ryul;Jeong, Eun-Taik
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.6
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    • pp.1285-1295
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    • 1997
  • Background : EGFR is one of the initial step in signal transduction pathway about multistep carcinogenesis. It is homologous to oncogene erbB-2 and is the receptor for EGF and TGF alpha. EGFR has important role in the growth and differentiation of tumor cells. So, EGFR in non-small cell lung cancer was examined to search for possible evidence as clinical prognostic factor. Methods : To investigate the role of EGFR in lung cancer, the author performed immunohistochemical stain of EGFR on 57 resected primary non-small cell lung cancer specimens. And the author analyzed the correlation between EGFR expression, clinical parameters, Sand $G_1$ phase fraction and survival. Results : 1) EGFR were detected in 56% of total 57 patients (according to histologic type, squamous cancer 50%, adenocarcinoma 63%, large cell cancer 75%) (according to TNM stage, stage I 64%, stage II 38%, stage III 55%) (according to cellular differentiation, well 50%, moderately 52%, poorly 65%). All differences were insignificant 2) Using the flow cytometric analysis, mean S-phase fraction of EGFR (+) and (-) group were 22.3(${\pm}10.5$)%. 18.0(${\pm}10.9$)% (p>0.05), mean $G_1$-phase fraction of EGFR (+) and (-) group were 68.4(${\pm}11.6$)%, 71.1(${\pm}12.8$)%, (p>0.05) 3) Two-year survival rate of EGFR (+) and (-) group were 53%, 84%, median survival time of EGFR (+) and (-) group were 26, 53 months. (p<0.05, Kaplan-Meier, generalized Wilcox) Conclusion : EGFR immunostaining may be a simple and useful method for survival prediction in non-small cell lung cancer.

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Role of HOXA Gene in Human Endometrial Decidualization (인간 자궁내막의 탈락막화에서 HOXA10 유전자의 역할)

  • Lee, Chang-Se;Park, Dong-Wook;Park, Chan-Woo;Kim, Tae-Jin
    • Clinical and Experimental Reproductive Medicine
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    • v.37 no.3
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    • pp.207-216
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    • 2010
  • Objective: This study was performed to clarify the role of HomeoboxA (HOXA) and its related signaling molecules in the decidualization of primary cultured endometrial cells. Methods: Human endometrial tissues were obtained by curettage of hysterectomy specimens from patients with conditions other than endometrial diseases. Tissues were minced and digested with Trypsin-EDTA for 20 min, $37^{\circ}C$. Cells were cultured with DMEM/F12 medium in $37^{\circ}C$, 5% $CO_2$ incubator for 24 hrs. Cells were treated with HOXA10 siRNA and added transforming growth factor (TGF)-${\beta}1$ (10 ng/mL) for 48 hrs to induces decidualization in vitro. Reverse transcription polymerase chain reaction analysis was accomplished to observe the expression of HOXA10, prolactin, cyclooxygenase (COX)-2, peroxisome proliferatoractivated receptor (PPAR)-$\gamma$, and wingless-type MMTV integration site family (Wnt). Results: HOXA10 expression was increased (1.8 fold vs. non-treated control) in TGF-${\beta}1$ treated cells. Decidualization marker, prolactin, was significantly increased in TGF-${\beta}1$ treated cells compared with HOXA10 siRNA treated cells. Endometrial cell differentiation marker, COX-2 was down-regulated by HOXA10 siRNA even if cells were treated with TGF-${\beta}1$. Wnt4 was down-regulated by treated with HOXA10 siRNA, this expression patters was not changed by TGF-${\beta}1$. Expression of PPAR-$\gamma$ was down regulated by TGF-${\beta}1$ in regardless of HOXA10 siRNA treatment. Conclusion: TGF-${\beta}1$ which is induced by progesterone in endometrial epithelial cells may induces stromal cell decidualization via HOXA10 and Wnt signaling cascade.

Christian Educational Proposals for Revitalizing Research on North Korea's 'education' (북한 '교육' 연구 활성화를 위한 기독교교육적 제언)

  • Ham, Seung su
    • Journal of Christian Education in Korea
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    • v.71
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    • pp.305-340
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    • 2022
  • This study was started to suggest the direction of Christian educational development to revitalize North Korea's 'education' research. Since the two Koreas have experienced heterogeneity in almost all elements of society, such as politics, economy, society, culture, and education, during the period of division in 1977, true unification depends on laying the foundation for social integration that can overcome the sense of heterogeneity between the two Koreas. This is why North Korea's "education" research is needed. Education is the foundation for transferring culture and history, and for bringing about the survival, transformation, and community of society and since it is the mission of Korean churches and Christian educators to establish the direction of North Korean "education" research, North Korean "education" research is very important. Despite this importance, 'North Korean research' in the field of Christian education has not been properly conducted. Research on the "Christian Unification Education Program" that can be used in churches is actively taking place, but research on the macro level of presenting post-unification education blueprints is rare. This study was started to suggest the direction of Christian educational development to revitalize North Korea's 'education' research. For the study, the characteristics of 'North Korea Research' were analyzed according to generational classification. As a result of the study, recent research on North Korea has been expanding in research topics and methodologies, and recent studies have been differentiated into microscopic studies that deviate from existing research trends at a macro level and view North Korea's daily life. The characteristics of 'North Korean education research' are summarized by period. The research on North Korean education, which began in earnest in the 1970s, was divided into the period of start(70s), transition(80s), leap(90s), expansion(2000s), and development(2010s~). and research characteristics for each period were analyzed. Through this, early North Korean education research was also conducted in the policy aspect of the country, and the characteristics of political and social studies were strong, but recent studies have confirmed that the subjects and contents are diversifying. Based on these studies, the pending issues and issues of North Korean education research in the field of Christian education were analyzed. The study of North Korea in the field of Christian education, which began in the 1980s, has been conducted in the engineering aspect of 'development of unification education programs for churches'. However, studies on Christian unification education and North Korean education itself, which can be used in public education sites including Christian schools, have yet to be sufficient. Nevertheless, the diversification of research in the field of Christian education can be evaluated as a positive change. Based on these studies, it was proposed to establish a de-ideological research foundation, secure primary research data(Raw Data), activate research topics and research methodologies, and strengthen research capabilities in the direction of development to revitalize North Korean research in the field of Christian education. I hope this study will trigger various follow-up studies and help Korean churches that must achieve unification.

Segmentation of the Cosmetics Market according to Shopping Orientation (구매성향에 따른 화장품시장 세분화)

  • Hwang Choon Sup
    • Journal of the Korean Society of Clothing and Textiles
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    • v.28 no.12 s.138
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    • pp.1632-1643
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    • 2004
  • The competitiveness of the domestic cosmetics market will improve more rapidly when efficient market segmentation is secured, qualitative information about each of the segmented markets is gathered, and adequate marketing strategies to meet the demands of each segmented markets is established. Therefore, continuous studies are needed to secure qualitative information for market segmentation. In order to find out whether the domestic cosmetics market could be segmented according to consumers' shopping orientation, the present study analyzed cosmetics purchasing behavior, complaint behavior, and demographic characteristics of each consumer group classified according to their shopping orientation. The study was implemented by a normative-descriptive survey method using a questionnaire. Three groups of consumers were identified according to their shopping orientation: price-oriented group, information-oriented group, and brand-oriented group. It was confirmed that the groups, classified according to shopping orientation, have significantly different demographic, purchasing, and complaint behavior. Therefore, shopping orientation should be carefully considered in the segmentation of the cosmetics market.

A New Efficient Cryopreservation of Human Embryonic Stem Cells by a Minimum Volume Cooling Method (Minimum Volume Cooling 방법을 이용한 효율높은 인간배아줄기세포동결)

  • Kim, Eun-Young;Park, Sae-Young;Yoon, Ji-Yeon;Ghil, Gwang-Su;Lee, Chang-Hyun;Lee, Gun-Soup;Tae, Jin-Cheol;Kim, Nam-Hyung;Lee, Won-Don;Chung, Kil-Saeng;Park, Se-Pill;Lim, Jin-Ho
    • Clinical and Experimental Reproductive Medicine
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    • v.31 no.1
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    • pp.41-50
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    • 2004
  • 연구 목적: 본 연구는 인간배아줄기세포 동결에 minimum volume cooling (MVC) 초자화 동결방법이 유용하게 이용될 수 있는지의 여부를 조사하고자 실시하였다. 연구재료 및 방법: 인간배아줄기세포 콜로니는 0.05% collagenase 처리와 기계적 처리에 의해 작은 clumps로 자른 다음 동결 방법에 따른 효율을 비교 검토하고자, i) 대략 40-50개의 clumps를 10% DMSO가 들어있는 동결액에 $5{\sim}10$분간 처리하여 1ml cryo-vial에 넣고 slow-cooling용 cryo-module에 장착하고 -80C에서 overnight한 후 $LN_2$에 침지하여 완만동결을 실시하거나 ii) 10% ethylene glycol (EG)이 들어 있는 동결액에서 5$\sim$10분 처리하고 30% EG과 0.5 mol sucrose가 들어 있는 동결액에서 30초간 처리하여 본 연구를 위해 개발된 MVC straw에 10 clumps씩 적재한 다음 $4{\sim}5$ MVC straw를 $LN_2$가 들어있는 cryo-vial에 넣어 MVC 초자화동결을 실시하였다. 융해 후 생존율을 조사하였고 배아줄기세포의 특성을 유지하고 있는지의 여부를 조사하였다. 결 과: 융해 후, 인간배아줄기세포의 생존율은 완만동결을 실시했던 군 (20.0%) 보다 MVC 초자화동결을 실시했던 군에서 (76.0%) 유의하게 높게 나타났다. 또한 회복과정에서 완만동결을 실시했던 군에서는 세포성장이 매우 더디고 안 좋은 반면, MVC 초자화동결을 실시했던 군은 배아줄기세포의 증식이 동결을 실시하지 않은 세포와 같은 상태로 2주 이후부터 빠르게 전환되고 회복되는 것을 확인할 수 있었다. 이와 더불어 MVC 초자화동결-융해 후 회복된 배아줄기세포에서 정상 핵형, alkaline phosphatase acitivity, SSEA-4와 TRA-1-60 염색 및 Oct-4 발현을 확인하였으며 체외분화의 특성도 확인하였다. 결 론: 새로이 개발된 MVC 초자화동결을 이용하면 인간배아줄기세포는 고유의 특성을 잃지 않고 성공적으로 동결될 수 있다.