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Studies on Developing Direct Gene Transfer Based on Naked Plasmid DNA for Treating Anemia (Naked Plasmid DNA를 이용한 빈혈 치료용 Direct Gene Transfer 시스템의 개발에 대한 연구)

  • Park Young Seoub;Jung Dong Gun;Choi Cha Yong
    • KSBB Journal
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    • v.19 no.5
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    • pp.341-347
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    • 2004
  • Several gene delivery therapies are being developed for treatment of serum protein deficiency. EPO is one of the most promising therapeutic agent for this treatment which is currently being investigated in depth. This study has the ultimate purpose of improving the gene delivery system for an increase of red blood cell production. A plasmid DNA was constructed smaller than other plasmids for an increase in penetration into animal cells, and two genes were cloned into each vector as a co-delivery system to express erythropoietin, and interluekin-3 or thrombopoietin, which can act on erythroid cell, thus activating hematopoiesis synergically. This co-delivery system has an advantage of decreasing the labour required for industrial production of DNA vaccine. A new plasmid vector, pVAC, in size 2.9 kb, was constructed with the essential parts from PUC 19 and pSectagB, which is much smaller than other plasmid vector and is the size of 2.9 kb. Co-delivery system was constituted by cloning human erythropoietin with each of human interluekin-3 gene or human thrombopoietin gene into both pVAC and pSectagB. As a result, the transfection efficiency of pVAC was higer than that of pSectagB in vitro, and hematocrit level of the mice injected with pVAC is higher than that of other mice. And co-delivery system, made of several plasmid DNAs, was expressed in vitro.

Immunoradiometric Assay using Monoclonal Antibody Against Human Serum Transferrin Receptor for Diagnosis of Iron Deficiency (사람 혈청 트란스페린수용체의 단클론 항체를 이용한 방사면역측정과 철영양상태의 진단)

  • 김승렬
    • Journal of Nutrition and Health
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    • v.29 no.9
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    • pp.971-980
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    • 1996
  • The soluble transferrin receptor(TfR) in human serum has been shown recently to be a truncated form of intact membrane bound receptor containing most of the extracellular domain. We purfied the transferin-free TfR from human serum by immounoaffinity chromatography which produced the single protein identity in high resolution gel chormatography. The monoclonal antibodies(MAb) against purifed serum TfR were produced by fusion of spleen cells o fimmunized Balb/c mice and SP2 cells. Ten hybrids producing MAb specific for serum TfR were identifed and determine their iostypes. A immunoraddiometric assay (IRMA) for serum TfR was established using two monoclonal IgG1 antibodies as the coating and indicator antibodies on the bosis of their suitability in sandwich IRMA of serum TfR. The mean serum TfR levels in the 15 normal male, 15 normal female, and 19 iron-deficient subjects were 5.4$\pm$0.98, 4.6$\pm$0/76, and 18.0$\pm$12.8mg/1, respectively, and the difference in mean values between normal and iron deficient subjects was significant(p=0.0005). There existed the inverse logarithmic relationship(r=-0.9336, p<0.0001) between the serum TfR and ferritin levels.

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Optimization and Efficiency Improvement of BCSC Solar Cells Using $MgF_{2}/CeO_{2}$Double Layer Antireflection Coatings ($MgF_{2}/CeO_{2}$ 이중반사방지막을 이용한 BCSC태양천지의 효율향상과 최적화)

  • 이욱재;임동건;이준신
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2001.07a
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    • pp.251-254
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    • 2001
  • This paper describes an efficiency improvement of buried contact solar cell (BSCS) with a structure of MgF$_2$/CeO$_2$/Ag/Cu/Ni grid/n$^{+}$ emitter/p-type Si base/p$^{+}$/Al. Theoretical and experimental investigations were performed on a double layer antireflection (DLAR) coating of MgF$_2$/CeO$_2$. We investigated CeO$_2$ films as an AR layer because they have a proper refractive index of 2.46 and demonstrate the same lattice constant as Si substrate. An optimized DLAR coating shewed a reflectance as low as 2.04 % in the wavelengths ranged from 0.4 ${\mu}{\textrm}{m}$ to 1.1 ${\mu}{\textrm}{m}$. BCSC cell efficiency was improved from 16.2 % without any AR coating to 19.9 % by employing DLAR coatings. Further details on MgF$_2$/CeO$_2$ DLAR coatings on the BCSC cells are presented in this paper.per.

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Immunostimulating Effects of Angelica Gigas by Radio-Protective (당귀의 방사선 방호에 의한 생체면역 증진 효과)

  • Kim Kyung-Yoon;Jeong Hyun-Woo;Kim Gye-Yeop
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.19 no.5
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    • pp.1256-1260
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    • 2005
  • The purpose of this study was to investigate the effects of Angelica gigas on jejunal survival, endogenous spleen colony formation and jejunal crypt cells of mice irradiated with Gamma-ray irradiation. The subject of this study includes 42 mice which were divided into each 7 groups. Angelica gigas experiment groups were Angelica gigas + Gamma-ray(10Gy), Angelica gigas + Gamma-ray(3Gy), Angelica gigas. Gamma-ray(1 Gy), Gamma-ray control (10Gy), Gamma-ray control(3Gy), Gamma-ray control(1Gy), Normal groups. In the present study to evaluate the effect of Angelica gigas on jejunal crypt survival, endogenous spleen colony formation, and apoptosis in jejunal crypt cells of mice Gamma-ray with each dose of Gamma-ray irradiation. The results of this study were as follows: In low-dose(1Gy) Gamma-ray radiation were treatment of Angelica gigas showed significantly increased(p<0.05) on the cell death apoptosis in crypt, intestine crypts survival of intestine after gamma-ray irradiation. High-dose(10Gy) Gamma-ray, treatment of Angelica gigas showed significantly increased(p<0.05) on the leukocyte. The above results suggest that Angelica gigas were immunostimulating effectively reduced Gamma-ray irradiation.

Antifungal Activity of the Crude Extract from Quintinia acutifolia on the Dermatophytic fungus

  • Lee, Jae-Sook;Chung, Jong-Gab;Oh, Hyun-Ju;Na, Young-Soon;Baek, Seung-Hwa
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.19 no.2
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    • pp.508-510
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    • 2005
  • The crude extract of Quintinia acutifolia Kirk inhibited the growth of the Gram positive bacterium Bacillus subtilis ATCC 19659, (3 mm inhibition zone at $150\;{\mu}g/disc$) and the dermatophytic fungus Trichophyton mentagrophytes ATCC 28185, (3 mm inhibition zone at $150\;{\mu}g/disc$), and cytotoxic to P388 murine leukaemia cells ATCC CCL 46 P388D1, ($IC_{50}$ $50,000\;{\mu}g/mL$ at $150\;{\mu}g/disc$). However, Candida albicans (ATCC 14053) did not observed the antimicrobial activity and the cytotoxic activity to BSC monkey kidney cells ($({\alpha})$ 5 mg/mL, $150\;{\mu}g/disc$).

Alteration in Leukocyte Subsets and Expressions of FcγR and Complement Receptors among Female Ragpickers in Eastern India

  • Mondal, Nandan K.;Siddique, Shabana;Banerjee, Madhuchanda;Roychoudhury, Sanghita;Mukherjee, Sayali;Slaughter, Mark S.;Lahiri, Twisha;Ray, Manas R.
    • Safety and Health at Work
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    • v.8 no.2
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    • pp.198-205
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    • 2017
  • Background: There are a million ragpickers in India who gather and trade recyclable municipal solid wastes materials for a living. The objective of this study was to examine whether their occupation adversely affects their immunity. Methods: Seventy-four women ragpickers (median age, 30 years) and 65 age-matched control housemaids were enrolled. Flow cytometry was used to measure leukocyte subsets, and leukocyte expressions of $Fc{\gamma}$ receptor I (CD64), $Fc{\gamma}RIII$ (CD16), complement receptor 1 (CD35) and CR3 (CD11b/CD18), and CD14. Serum total immunoglobulin-E was estimated with enzyme-linked immunosorbent assay. Results: Compared with the controls, ragpickers had significantly (p < 0.0001) higher levels of CD8-T-cytotoxic, CD16+CD56+natural killer, and CD4+CD45RO+memory T-cells, but depleted levels of CD19+B-cells. The percentage of CD4+T-helper-cells was lower than the control group (p < 0.0001), but their absolute number was relatively unchanged (p = 0.42) due to 11% higher lymphocyte counts in ragpickers. In ragpickers, the percentages of CD14+CD16+intermediate and CD14dim CD16+nonclassical monocyte subsets were elevated with a decline in CD14+CD16-classical monocytes. The expressions of CD64, CD16, CD35, and CD11b/CD18 on both monocytes and neutrophils, and CD14 on monocytes were significantly higher in ragpickers. In addition, ragpickers had 2.7-times more serum immunoglobulin-E than the controls (p < 0.0001). After controlling potential confounders, the profession of ragpicking was positively associated with the changes. Conclusion: Ragpicking is associated with alterations in both innate (neutrophils, monocytes, and natural killer cell numbers and expression of complement and $Fc{\gamma}$ receptors) and adaptive immunity (numbers of circulating B cells, helper, cytotoxic, and memory T cells).

Reduction of Mitochondrial Derived Superoxide by Mito-TEMPO Improves Porcine Oocyte Maturation In Vitro (Mito-TEMPO에 의한 미토콘드리아 유래 초과산화물의 감소가 돼지 난모세포 성숙에 미치는 영향)

  • Yang, Seul-Gi;Park, Hyo-Jin;Lee, Sang-Min;Kim, Jin-Woo;Kim, Min-Ji;Kim, In-Su;Jegal, Ho-Geun;Koo, Deog-Bon
    • Journal of Animal Reproduction and Biotechnology
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    • v.34 no.1
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    • pp.10-19
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    • 2019
  • Morphology of cumulus-oocyte-complexes (COCs) at germinal vesicle (GV) stage as one of the evaluation criteria for oocyte maturation quality after in vitro maturation (IVM) plays important roles on the meiotic maturation, fertilization and early embryonic development in pigs. When cumulus cells of COCs are insufficient, which is induced the low oocyte maturation rate by the increasing of reactive oxygen species (ROS) in porcine oocyte during IVM. The ROS are known to generate including superoxide and hydrogen peroxide from electron transport system of mitochondria during oocyte maturation in pigs. To regulate the ROS production, the cumulus cells is secreted the various antioxidant enzymes during IVM of porcine oocyte. Our previous study showed that Mito-TEMPO, superoxide specific scavenger, improves the embryonic developmental competence and blastocyst formation rate by regulating of mitochondria functions in pigs. However, the effects of Mito-TEMPO as a superoxide scavenger to help the anti-oxidant functions from cumulus cells of COCs on meiotic maturation during porcine oocyte IVM has not been reported. Here, we categorized experimental groups into two groups (Grade 1: G1; high cumulus cells and Grade 2: G2; low cumulus cells) by using hemocytometer. The meiotic maturation rate from G2 was significantly (p < 0.05) decreased (G1: $79.9{\pm}3.8%$ vs G2: $57.5{\pm}4.6%$) compared to G1. To investigate the production of mitochondria derived superoxide, we used the mitochondrial superoxide dye, Mito-SOX. Red fluorescence of Mito-SOX detected superoxide was significantly (p < 0.05) increased in COCs of G2 compared with G1. And, we examined expression levels of genes associated with mitochondrial antioxidant such as SOD1, SOD2 and PRDX3 using a RT-PCR in porcine COCs at 44 h of IVM. The mRNA levels of three antioxidant enzymes expression in COCs from G2 were significantly (p < 0.05) lower than COCs of G1. In addition, we investigated the anti-oxidative effects of Mito-TEMPO on meiotic maturation of porcine oocyte from G1 and G2. Meiotic maturation and mRNA levels of antioxidant enzymes were significantly (p < 0.05) recovered in G2 by Mito-TEMPO ($0.1{\mu}M$, MT) treatment (G2: $68.4{\pm}3.2%$ vs G2 + MT: $73.9{\pm}1.4%$). Therefore, our results suggest that reduction of mitochondria derived superoxide by Mito-TEMPO may improves the meiotic maturation in IVM of porcine oocyte.

Comparative Characteristics of Three Human Embryonic Stem Cell Lines

  • Lee, Jung Bok;Kim, Jin Mee;Kim, Sun Jong;Park, Jong Hyuk;Hong, Seok Ho;Roh, Sung Il;Kim, Moon Kyoo;Yoon, Hyun Soo
    • Molecules and Cells
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    • v.19 no.1
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    • pp.31-38
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    • 2005
  • Human embryonic stem (hES) cells have unique features including unlimited growth capacity, expression of specific markers, normal karyotypes and an ability to differentiate. Many investigators have tried to use hES cells for cell-based therapy, but there is little information about the properties of available hES cell lines. We compared the characteristics of three hES cell lines. The expression of SSEA-1, -3, -4, and APase, was examined by immunocytochemistry, and Oct-4 expression was analyzed by RT-PCR. Differentiation of the hES cells in vitro and in vivo led to the formation of embryoid bodies (EBs) or teratomas. We examined the expression of tissue-specific markers in the differentiated cells by semiquantitative RT-PCR, and the ability of each hES cell line to proliferate was measured by flow cytometry of DNA content and ELISA. The three hES cell lines were similar in morphology, marker expression, and teratoma formation. However there were significant differences (P < 0.05) between the differentiated cells formed by the different cell lines in levels of expression of tissue-specific markers such as renin, kallikrein, Glut-2, ${\beta}-$ and ${\delta}-globin$, albumin, and ${\alpha}1-antitrypsin$ (${\alpha}1-AT$). The hES cell lines also differed in proliferative activity. Our observations should be useful in basic and clinical hES cell research.

Immunological Characteristics of Hyperprogressive Disease in Patients with Non-small Cell Lung Cancer Treated with Anti-PD-1/PD-L1 Abs

  • Kyung Hwan Kim;Joon Young Hur;Jiae Koh;Jinhyun Cho;Bo Mi Ku;June Young Koh;Jong-Mu Sun;Se-Hoon Lee;Jin Seok Ahn;Keunchil Park;Myung-Ju Ahn;Eui-Cheol Shin
    • IMMUNE NETWORK
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    • v.20 no.6
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    • pp.48.1-48.11
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    • 2020
  • Hyperprogressive disease (HPD) is a distinct pattern of progression characterized by acceleration of tumor growth after treatment with anti-PD-1/PD-L1 Abs. However, the immunological characteristics have not been fully elucidated in patients with HPD. We prospectively recruited patients with metastatic non-small cell lung cancer treated with anti-PD-1/PD-L1 Abs between April 2015 and April 2018, and collected peripheral blood before treatment and 7-days post-treatment. HPD was defined as ≥2-fold increase in both tumor growth kinetics and tumor growth rate between pre-treatment and post-treatment. Peripheral blood mononuclear cells were analyzed by multi-color flow cytometry to phenotype the immune cells. Of 115 patients, 19 (16.5%) developed HPD, 52 experienced durable clinical benefit (DCB; partial response or stable disease ≥6 months), and 44 experienced non-hyperprogressive progression (NHPD). Patients with HPD had significantly lower progression-free survival (p<0.001) and overall survival (p<0.001). When peripheral blood immune cells were examined, the pre-treatment frequency of CD39+ cells among CD8+ T cells was significantly higher in patients with HPD compared to those with NHPD, although it showed borderline significance to predict HPD. Other parameters regarding regulatory T cells or myeloid derived suppressor cells did not significantly differ among patient groups. Our findings suggest high pre-treatment frequency of CD39+CD8+ T cells might be a characteristic of HPD. Further investigations in a larger cohort are needed to confirm our results and better delineate the immune landscape of HPD.

Effect of Aspirin on the Acute Lung Injury Induced by Intestinal Ischemia/Reperfusion. (장의 허혈-재관류로 유도된 급성 폐손상에서 아스피린의 작용)

  • Park, Yoon-Yub
    • Journal of Life Science
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    • v.19 no.6
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    • pp.818-824
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    • 2009
  • The mechanisms responsible for ischemia/reperfusion (I/R) injury have direct or indirect relevance to clinical lung injury after severe shock, cardiopulmonary bypass, and transplantation. This study investigated the effects of aspirin on intestinal I/R-induced acute lung injury (ALI) in rats. Lipopolysaccharide (LPS) induced cyclooxygenase-2 (COX-2) expression in A549 and RAW264.7 cells. RAW264.7 macrophages had shown greater expression of COX-2 than A549 cells. In addition, the NADPH oxidase inhibitor apocynin and p38 MAPK inhibitor SB203580 attenuated LPS-stimulated COX-2 expression. To induce ALI, intestinal ischemia was performed for 60 min prior to the 4 hr reperfusion by clamping the superior mesenteric artery in Sprague-Dawley rats. In order to test and compare the effect of non-specific COX inhibitor aspirin with the effect of mepacrine, a well known phospholipase$A_{2}$ inhibitor, rats were divided into 4 groups: Sham, I/R, Mepa+I/R (mepacrine, 60 mg/kg, i.p.), ASA+I/R (aspirin, 10 mg/kg, i.p.). In the present investigation, myeloperoxidase activities in the lung and intestinal tissues were increased by I/R. These changes were reduced by single pretreatment of mepacrine (60 mg/kg, i.p.) or aspirin (10 mg/kg, i.p.) 30 min before I/R. Structural studies demonstrated that the tissue injuries in the lung and intestine after I/R were also attenuated by the pretreatment of mepacrine or aspirin. These results suggest that I/R-induced ALI is mediated, in part, by the activation of COX. In addition, pretreatment of aspirin might be helpful for the prevention of ALI in ARDS-prone patients. In addition, the p38 MAPK inhibitor and apocynin also might be helpful to ALI through the inhibition of COX-2 expression.