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Post-Activation Treatment with Cytochalasins and Latrunculin A on the Development of Pig Oocytes after Parthenogenesis and Somatic Cell Nuclear Transfer

  • Park, Bola;Lee, Joohyeong;Lee, Yongjin;Elahi, Fazle;Jeon, Yubyeol;Hyun, Sang-Hwan;Lee, Eunsong
    • Journal of Embryo Transfer
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    • v.28 no.2
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    • pp.133-139
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    • 2013
  • The objective of this study was to determine the effect of post-activation treatment with cytoskeletal regulators in combination with or without 6-dimethylaminopurine (DMAP) on embryonic development of pig oocytes after parthenogenesis (PA) and somatic cell nuclear transfer (SCNT). PA and SCNT oocytes were produced by using in vitro-matured pig oocytes and treated for 4 h after electric activation with $0.5{\mu}M$ latrunculin A (LA), $10.4{\mu}M$ cytochalasins B (CB), and $4.9{\mu}M$ cytochalasins D (CD) together with none or 2 mM DMAP. Post-activation treatment of PA oocytes with LA, CB, and CD did not alter embryo cleavage (85.8~88.6%), blastocyst formation (30.7~ 32.4%), and mean cell number of blastocysts (33.5~33.8 cells/blastocyst). When PA oocytes were treated with LA, CB, and CD in combination with DMAP, blastocyst formation was significantly (P<0.05) improved by CB+DMAP (42.5%) compared to LA+DMAP (28.0%) and CD+DMAP (25.1%), but no significant differences were found in embryo cleavage (77.5~78.0%) and mean blastocyst cell number (33.6~35.0 cells) among the three groups. In SCNT, blastocyst formation was significantly (P<0.05) increased by post-activation treatment with LA+DMAP (32.9%) and CD+DMAP (35.0%) compared to CB+DMAP (22.0%) while embryo cleavage (85.5~85.7%) and blastocyst cell number (41.1~43.8 cells) were not influenced. All three treatments (LA, CB, and CD with DMAP) effectively inhibited pseudo-polar body extrusion in SCNT oocytes. The proportions of oocytes showing single pronucleus formation were 89.6%, 83.9%, and 93.3%, respectively with the increased tendency (P<0.1) by LA+DMAP and CD+ DMAP compared to CB+DMAP. Our results demonstrate that post-activation treatment with LA or CD in combination with DMAP improves pre-implantation development of SCNT embryos and the stimulating effect of cytoskeletal modifiers on embryonic development is differentially shown depending on the origin (PA or SCNT) of embryos in pigs.

Leukocyte Markers Differentiate Non-Infected from Spontaneously Infected Dairy Cows (우유의 체세포내 면역 표지자 분석을 통한 소 유방염 진단)

  • Yu, Do-Hyeon;Lee, Jong-Hyeon;Song, Ru-Hui;Noh, Dong-Ho;Li, Ying-Hua;Lee, Mi-Jin;Park, Jin-Ho
    • Journal of Veterinary Clinics
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    • v.26 no.6
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    • pp.524-527
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    • 2009
  • Spontaneously infected and non-infected dairy cows were assessed in a cross-sectional study aimed at determining whether bovine leukocyte markers may diagnose intra-mammary infections (bovine mastitis). Animals located in herds where bovine mastitis was highly prevalent were investigated (n = 31 animals). The expression of three cell-surface markers (CD11b, CD4 and CD8) was assessed, and the somatic cell count (SCC) and bacteriological analyses (both cultures and PCR tests) were also conducted. Cows identified as infected revealed statistically significant higher milk leukocyte CD11b, CD4 percentage and milk CD4/CD8 ratios than non-infected cows. Immunological markers may diagnose spontaneous bovine mastitis.

Effects of Partial Cervus elaphus Linne' Extract on Antitumoral Immune Response in Melanoma-induced Mice (부위별(部位別) 녹용(鹿茸)이 흑색종(黑色腫) 유발(誘發) 생쥐의 종양억제(腫瘍抑制)와 면역기능(免疫機能)에 미치는 영향(影響))

  • Oh Ha-Souk;Kim Jang-Hyun
    • The Journal of Pediatrics of Korean Medicine
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    • v.14 no.1
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    • pp.39-78
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    • 2000
  • Introduction: In order to investigate the effects of partial Cervus elaphus Linne' extract on antitumoral immunological, a group of mice are melanoma-induced and observed reponses in terms of the number of lymphocyte, CD4+ count, CD8+ count, CD4+/CD8+ ratio in blood and spleen, change in body weight, melanoma weight, spleen index, NK cell activity and productivity of IL-2 in each mouse Methodology: Male C57BL/6 mice were chosen as an experimental object and were divided into 5 groups randomly selection. The normal group did not receive any induction. The control group was treated with normal saline in melanoma-induced' mice. Sample I group induced the upper part of Cervus elaphus Linne' extract in melanoma-induced mice. Sample II group was induced the middle part of Cervus elaphus Linne' extract in melanoma-induced mice. Sample III group was induced the lower part of Cervus elaphus Linne' extract in melanoma-induced mice. The dosage of medication was 0.2cc daily for 14days. Results: 1. There was a significant difference in the number of lymphocyte in spleen in the sample I (upper part of Cervus elaphus Linne' extract induced) and the sample II (middle part of Cervus elaphus Linne' extract induced) compared to the control group and the sample III (lower part of Cervus elaphus Linne' extract induced). On the other hand, no significant difference was observed in the number of lymphocyte in blood in the control group and all sample groups. 2. In the CD4+ T cell ratio in blood, all three sample groups showed differences compared to the control group, though there was no significant difference between sample groups. In the CD4+ T cell ratio in spleen, there was a difference between the sample I and the control group, while the sample II and the sample III had significant difference to the control group. And also, it has been observed there were differences between the sample I and the other samples. 3. In the CD8+ T cell ratio in spleen, all three sample groups showed significant differences compared to the control group, while there was no difference between groups in the ratio in blood. 4. In the CD4+/CD8+ T cell ratio in blood, the sample I showed a significant difference compared to the control group, while the sample 1I and sample III showed differences compared to the control group. In the CD4+/CD8+ T cell ratio in spleen, all three samples showed a significant difference compared to the control group, when the sample I had a difference to the other sample groups. 5. The spleen index of the sample I and the sample II showed a significant difference compared to the sample III and the control group. In comparison between the sample groups, the sample I and the sample II showed a significant difference to the sample III. 6. In terms of the change in body weight and melanoma weight, all three sample groups showed a significant difference compared to the control group, while the comparison between the sample groups showed the sample and the sample II had a significant difference to the sample Ⅲ. 7. In comparison of NK cell activity, the sample I had a difference compared to the other groups when the effector to target cell ratio was 2.5:1. With the ratio of 5:1, the sample I and sample II showed significant differences compared to the control group, while the sample Ⅲ showed a difference. When the effector to target cell ratio was 10:1, there was no difference between groups. 8. In the productivity of IL-2, all three sample groups showed significant differences compared to the control group. In comparison between sample groups, there were significant differences between each sample groups in order of the sample I , the sample II and the sample Ⅲ. Conclusion: As one can witness from the above results, administration of partial Cervus elaphus Linne extract played important role in antitumoral immune response in melanoma-induced mice, and it could be suggested that sample I and sample II groups have prominent antitumoral immune effect.

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Characterization of Human Dental Pulp Cells from Supernumerary Teeth by Using Flow Cytometry Analysis (유세포 분석을 통한 과잉치 치수 유래 세포의 줄기세포 특성 연구)

  • You, Yonsook;Kim, Jongbin;Shin, Jisun;Lee, June-Haeng;Kim, Jongsoo
    • Journal of the korean academy of Pediatric Dentistry
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    • v.46 no.3
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    • pp.337-342
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    • 2019
  • The aim of this study was to analyze cells from human dental pulp tissue of impacted supernumerary teeth as stem cells with flow cytometry. Human dental pulp cells from 15 supernumerary teeth were identified their characteristics as stem cells by expression of mesenchymal stem cell markers through flow cytometry analysis at passage 3 and passage 10. Cluster of differentiation (CD) 73, CD 90, CD 34, CD 45 and STRO-1 cell surface markers were used to figure out characteristics of dental pulp stem cells from supernumerary teeth. At passage 3, the cell population showed positive expression of CD 73, CD90 and STRO-1, lacked expression of CD 34 and CD 45. At passage 10, CD 73, CD 90 and STRO-1 showed positive expression while CD 34 and CD 45 showed negative expression. This study indicated that dental pulp stem cells of supernumerary teeth had the properties of mesenchymal stem cells at both early and late passage. Impacted supernumerary teeth could be considered as a noble source of stem cells because of rapid growth and maintaining characteristics of stem cells until late passage.

Twist2 Regulates CD7 Expression and Galectin-1-Induced Apoptosis in Mature T-Cells

  • Koh, Han Seok;Lee, Changjin;Lee, Kwang Soo;Park, Eun Jung;Seong, Rho H.;Hong, Seokmann;Jeon, Sung Ho
    • Molecules and Cells
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    • v.28 no.6
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    • pp.553-558
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    • 2009
  • In the periphery, a galectin-1 receptor, CD7, plays crucial roles in galectin-1-mediated apoptosis of activated T-cells as well as progression of T-lymphoma. Previously, we demonstrated that $NF-{\kappa}B$ downregulated CD7 gene expression through the p38 MAPK pathway in developing immature thymocytes. However, its regulatory pathway is not well understood in functional mature T-cells. Here, we show that CD7 expression was downregulated by Twist2 in Jurkat cells, a human acute T-cell lymphoma cell line, and in EL4 cells, a mature murine T-cell lymphoma cell line. Furthermore, ectopic expression of Twist2 in Jurkat cells reduced galectin-1-induced apoptosis. While full-length Twist2 decreased CD7 promoter activity, a C-terminal deletion form of Twist2 reversed its inhibition, suggesting an important role of the C-terminus in CD7 regulation. In addition, CD7 expression was enhanced by histone deacetylase inhibitors such as trichostatin A and sodium butyrate, which indicates that Twist2 might be one of candidate factors involved in histone deacetylation. Based on these results, we conclude that upregulation of Twist2 increases the resistance to galectin-1-mediated-apoptosis, which may have significant implications for the progression of some T-cells into tumors such as Sezary cells.

Correlation Between Expression of Cell Adhesion Molecules CD44 v6 and E-cadherin and Lymphatic Metastasis in Non-small Cell Lung Cancer

  • Su, Chong-Yu;Li, Yun-Song;Han, Yi;Zhou, Shi-Jie;Liu, Zhi-Dong
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.5
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    • pp.2221-2224
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    • 2014
  • Objective: To explore the relationship between expressions of cell adhesion molecules CD44 v6 and E-cadherin (E-cad) and lymphatic metastasis in non-small cell lung cancer (NSCLC). Materials and Methods: Eightyseven tissue samples obtained from patients with primary NSCLC were collected in our hospital from Dec., 2007 to Dec., 2012, and the expressions of CD44 v6 and E-cad gene proteins in these samples were detected by immunohistochemical method. Results: In the tissue without lymphatic metastasis, the positive expression rate of CD44 v6 was significantly lower, whereas the normal expression rate of E-cad was notably higher than that with lymphatic metastasis (55.6% vs. 78.4%, 47.2% vs. 21.6%), and both differences had statistical significance (P<0.05). Besides, CD44 v6 and E-cad expressions had a significant correlation in the NSCLC tissue with lymphatic metastasis (P<0.05). Conclusions: The positive expression of CD44 v6 and abnormal expression of E-cad may play a very important role in promoting lymphatic metastasis of NSCLC, with synergistic effect. Hence, detection of CD44 v6 and E-cad expressions is conductive to judging the lymphatic metastasis in NSCLC.

Intracellular Signaling Pathways for Type II IgE Receptor (CD23) Induction by Interleukin - 4 and Anti - CD40 Antibody

  • Kim, Hyun-Il;Park, Hee-Jeoung;Lee, Choong-Eun
    • BMB Reports
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    • v.30 no.6
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    • pp.431-437
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    • 1997
  • Since the role of CD40 on the interleukin-4(IL-4) -induced B cell activation has been strongly implicated in the agumentation of IgE production and response, we have investigated the intracelluar signaling pathways utilized by IL-4 and CD40 for type II IgE receptor (CD23) expression. IL-4 and anti-CD40 antibody treatment of human B cells, independently caused a rapid induction of CD23 gene activation within 2 h. There was a noticeable synergism between the action of the two agents inducing CD23 expression: the addition of anti-CD40 to the IL-4-treated culture significantly agumented the IL-4-induced CD23 on both mRNA and surface protein levels, and the inclusion of IL-4 in the anti-CD40-treated cells caused a further increase of CD23 expression far above the maximal level induced by anti-CD40. Protein tyrosine kinase (PTK) inhibitors effectively suppressed the both IL-4- and anti -CD40-induced CD23 expression. whereas protein kinase C (PKC) inhibitors had no effects. Electrophoretic mobility shift assays (EMSA) have shown that IL-4 and anti-CD40 induce the activation of NF-IL-4 and $NF-_{K}B$, respectively, binding to the CD23 promoter, both in a PKC-independent and PTK-dependent manner. These data suggest that the synergistic activation of CD23 gene expression by IL-4 and anti-CD40 is mediated by co-operative action of distinct nuclear factors. each of which is rapidly activated via PKC-independent and PTK-dependent process.

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Immunostimulatory effect of Korean traditional medicine Acanthopanacis Cortex

  • Chang, In-Ae;Shin, Hye-Young;Kim, Youn-Chul;Yun, Yong-Gab;Park, Hyun
    • Natural Product Sciences
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    • v.13 no.4
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    • pp.283-288
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    • 2007
  • Acanthopanacis Cortex (AC) has been popularly used as an herbal medicine for medical treatment of rheumatoid arthritis, insomnia, impotence and diabetes. Here, we investigated immunostimulating effects of the aqueous extract of AC on macrophage. We studied nitric oxide (NO) and tumor necrosis factor (TNF)-${\alpha}$ release in response to AC treatment, as they are important secretory products of macrophage. AC alone induce the NO and TNF-${\alpha}$ production. AC increase c-Jun NH2-terminal kinase 1/2 (JNK) and extracellular signal-regulated kinase (ERK) phosphorylation but does not p38 activation in RAW 264.7 cells. Also AC resulted in the enhanced cell-surface expression of CD80 and CD14. In addition, AC resulted in enhanced T cell-stimulatory capacity and increased T cell secretion of interferon (IFN)-gamma. After feeding with AC to mouse for 10 days, the change of $CD28^+$ and $CD40^+$ population was analyzed. AC increased $CD28^+$ population in splenocytes in vivo. These studies indicate that AC induces macrophage activation and suggest the possible use of AC in macrophage-based immunotherapies.

Anti-arthritis Effects of Jeonsaenghwalhyeoltanggamibang through Immune Modulation Modulation (전생활혈탕가미방(全生活血湯加味方)의 면역조절 작용을 통한 항관절염 효능에 관한 연구)

  • Ko, Jeong-Min;Choe, Chang-Min;Cho, Han-Baek;Kim, Song-Baeg
    • The Journal of Korean Obstetrics and Gynecology
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    • v.22 no.2
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    • pp.1-25
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    • 2009
  • Purpose: The purpose of this study is to investigate the anti-arthritis effects of Jeonsaenghwalhyeoltanggamibang(JHTG) on collagen-induced arthritis(CIA) in mice. Methods: To assess the effects of JHTG on CIA in mice, we conducted several experiments such as analysis of arthritis index, cell count of draining lymph node(DLN) and paw joint, measurement of serum antibody levels and observation of the histological changes of joint. Results: 1. JHTG extract had a suppressive effect on the arthritis index of paw joints in CIA mice. 2. JHTG extract increased the total cell number of DLN, and decreased the total cell number of paw joints in CIA mice. 3. JHTG extract increased the absolute number of various cell surface receptors in DLN, and decreased the absolute number of B220+/CD23+ cells in DLN in CIA mice. 4. JHTG extract decreased the absolute number of CD3+, CD4+, CD11b+/Gr-1 cells in paw joint in CIA mice. 5. JHTG extract didn't decrease the absolute number of CD4+/CD25+ cells in paw joints in CIA mice. 6. JHTG extract decreased levels of total IgM in the serum of CIA mice, but had no effect on levels of collagen II specific antibody. 7. JHTG extract decreased the destruction of articular cartilages and collagen fibers and the proliferation of synovial cells in paw joints from CIA mice. Conclusion: These results indicate that JHTG has clinical potential for the treatment of rheumatoid arthritis by modulating the immune response.

Effects of Kamichihyo-san on Anti-CD40 and Recombinant Interleukin-4 Induced Cytokine Production and Immunoglobulin E in Highly Purified Mouse B Cells (생쥐의 B 세포에서 anti-CD40과 rIL-4로 유도된 사이토카인 생산과 면역글로블린 E에 대한 가미치효산의 효과)

  • Ham Chul in;Park Yang chun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.6
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    • pp.1479-1486
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    • 2003
  • In order to evaluate the antiallergic effects of Kamiohihyosan(KCHS), studies were done. We measured the cytotoxic activity for lung fibroblast cell, cytokines transcript expression, production of IL-4, IL-10, IFN-γ, proliferation of B cell in anti-CD40mAb plus rIL-4 stimulated murine splenic B cells. The results were obtained as follows: KCHS was not showed cytotoxicity in the fibroblast lung cell, KCHS increased the gene synthesis of INF-γ, TNF-α, IL1-β, IL-6, IL-10(m-RNA), KCHS decreased the gene synthesis of IL-4, IL-5, TGF-β(m-RNA), KCHS decreased the appearance of IL-4, IgE significantly, KCHS increased the appearance of IL-10, IFN-γ significantly, KCHS decreased the proliferation of B cell significantly, The facts above prove that KCHS is effective against the allergy. Thus, I think that we should study on this continuously.