• 한국동물학회:학술대회논문집
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• 한국동물학회 1995년도 한국생물과학협회 학술발표대회
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• pp.136.1-136
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• 1995

No Abstract, See Full Text

• Toxicological Research
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• 제18권1호
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• pp.79-85
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• 2002

YHB216 is one of new recombinant human erythropoietins (rHu-EPO) developed by Yuhan Research Institute. The rHu-EPO products are widely being used for the treatment of various types of anemia. As a series of safety studies on YHB216, we performed the local irritation test (dermal & ocular application) in male New Zealand White rabbits and micronucleus test in male ICR mice. In the skin irritation test, 0.5 ml of YHB216 10,000 IU/ml solution was applied to the back skin of rabbits for 24 hours and sub-sequent observation was performed. There was no induced response after the treatment and the primary irritation index (P.I.I.) was‘0’. In the eye irritation test, 0.1 ml of YHB216 10,000 IU/mL solution was instilled into the conjunctiva of the eye. No treatment-related reaction was observed at the cornea, iris, and conjunctiva. In the micronucleus test, YHB216 was administered intravenously to male mice (6 mice per group) at dose levels of 0, 6,250, 12,500, and 25,000 IU/kg. Bone marrow cells were collected at 24 hours after the treatment. YHB216 treated groups showed no significant difference in the P/N (polychromatic erythrocyte/ normochromatic erythrocyte) ratio and in the number of micronucleated polychromatic erythrocyte com-pared with the control. In conclusion, YHB216 was found to be a non-irritating material up to 10,000 IU/ml in the local irritation test and to be a non-mutagen up to 25,000 IU/kg in the micronucleus test.

• KSBB Journal
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• 제4권1호
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• pp.34-39
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• 1989

본 논문은 동물 세포의 대량 배양을 위해 연속 공법 방식인 Perfusion Continuous System을 도입해 의약적으로 중요한 EPO의 생산을 위한 생물 공학적인 자료들을 제고하고 있다. 이 System은 세포 증식 속도를 배지의 Perfusion 속도로 변화시킴으로써 조절시킬 수 있는 산소 소비속도와 밀접한 상관관계가 있음을 입증하며 이는 세포수의 직접 측정에 따른 오차 및 방법상의 문제를 정학히 측정할 수 있는 간접 변수, 즉 산소소비속도를 이용함으로써 제거할 수 있다. 특히 이 산소소비속도와 세포 성장 관계로 model로써 세포 증식을 예측함과 동시에 동물 세포 대량 배양을 위한 scale-up의 중요한 기초자료가 될 것이다. 지금까지 발표되지 않았던 동물 세포의 glucose에 대한 True growth yield와 maintenance coefficient값들의 측정은 동물 세포 성자관 유용 물질 생산을 위한 중요한 수율적 자료가 된다.한편 이 결과는 지금까지 미생물이나 광합성에서만 적용되었던 yield model이 Eukaryotes에서도 응용될 수 있음을 증명하고 있다. 이와 같이 perfusion system이 많은 장점을 갖고 있지만, 세포 성장에 따른 동력학적인 연구의 수행이 좀 더 요구되는 실정이며, 특히 Perfusion system을 설명할 수 있는 이론 및 cytostatic moel의 정립이 선행되어야 할 것이다.

• BMB Reports
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• 제32권2호
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• pp.112-118
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• 1999

The presence of hypoxic cells in solid tumors has long been considered a problem in cancer treatment such as in radiation therapy or treatment with some anticancer drugs. It has been suggested that hypoxic cells are involved in the development of a more aggressive phenotype and contribute to metastasis. In this study, as an attempt to understand how tumor cells adapt to hypoxic stress, we investigated the regulation of the hypoxia-induced expression of proteins that control essential processes of tumor cell survival and angiogenesis. We first examined whether hypoxia induces stress protein gene expression of murine solid tumor RIF cells. We also examined hypoxia-induced changes in angiogenic gene expression in these cells. Finally, we investigated the association of the elevated levels of stress proteins with the regulation of hypoxia-induced angiogenic gene expression. Results demonstrated that hypoxia induced the expression of the erythropoietin (EPO) gene and at least two major members of stress proteins, heat shock protein 70 (HSP70) and 25 (HSP25) in RIF tumor cells. Evidence that the expression of EPO gene was greatly potentiated in TR cells suggested that the elevated levels of HSPs may play an important role in the regulation of the hypoxia-induced EPO gene expression. One of the RIF variant cell lines, TR, displays elevated levels of HSPs constitutively. Taken together, our results suggest that a hypoxic tumor microenvironment may promote the survival and malignant progression of the tumor cells by temporarily increasing the level of stress proteins and expressing angiogenic genes. We suspect that stress proteins may be associated with the increase of the angiogenic potential of tumor cells under hypoxia.

• 생약학회지
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• 제27권4호
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• pp.371-377
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• 1996

In previous studies we reported that the levels of RBC, hemoglobulin (HGB) and hematocrit (HCT) in SAM P6 were increased significantly from 7 day after oral administration of the pilose antler extract, 5 g/kg/day, and were lasted during the study. Thus, this study was performed to elucidate mechanism of erythropoietic action by the extract administration. SAM R1 and SAM P6 were chosen as experimental animals. At age of 12 weeks, pilose antler extract were given 0.3 and 5 g/kg/day (p.o.) each for 0, 7 and 14 days in both animals. Complete blood cells such as RBC, HGB, and HCT were counted. And plasma concentration of erythropoietin (EPO) which is the major regulator of erythropoiesis was measured using $^{125}I-antierythropoietin$ IgG. Total iron concentration in plasma was also analyzed. The levels of RBC and HCT were increased significantly after administration at both doses of 0.3 and 5 g/kg/day in SAM P6, however, these were increased only at dose of 5 g/kg/day in SAM R1. The plasma EPO concentration was increased significantly after administration in SAM P6. The plasma concentrations of total iron were significantly decreased after administration of the extract in SAM P6. These results suggest that the changes in erythropoietic effects after the administration of pilose antler extract may be mediated, at least in part, through the change in the plasma EPO.

• 동의생리병리학회지
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• 제27권6호
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• pp.795-801
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• 2013

The purpose of this study is to investigate effects of White Ginseng-Ejung-tang (EG) and Red Ginseng-Ejung-tang (ER) water extract on production of various cytokines such as interleukin (IL)-21, IL-25, IL-$28{\beta}$, erythropoietin (EPO), Exodus-2, monocyte chemotactic protein (MCP)-5, macrophage inflammatory protein (MIP)-$3{\alpha}$, MIP-$3{\beta}$, Fractalkine, and TARC in RAW 264.7 mouse macrophages stimulated by lipopolysaccharide (LPS). Levels of cytokines were measured by High-throughput multiplex bead array cytokine assay based on xMAP (multi-analyte profiling beads) technology. ER significantly decreased levels of IL-21, IL-25, IL-$28{\beta}$, EPO, Exodus-2, MCP-5, MIP-$3{\alpha}$, MIP-$3{\beta}$, TARC, and fractalkine for 24 h incubation at the oncentrations of 25 and 100 ${\mu}g/mL$ in LPS-induced RAW 264.7 (P < 0.05). But EG did not show any significant effect. These results suggest that ER has anti-inflammtory property related with its inhibition on the production of IL-21, IL-25, IL-$28{\beta}$, and chemokines such as EPO, MCP-5, MIP-$3{\alpha}$, MIP-$3{\beta}$, Fractalkine, Exodus-2, and TARC in LPS-induced macrophages.

• BMB Reports
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• 제53권3호
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• pp.148-153
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• 2020

Erythropoietin and iron have individually shown beneficial effects on early-phase liver regeneration following partial hepatectomy (PHx); however, there are limited data on the combined effect on late-phase liver regeneration after PHx. Here we examined combined effects of recombinant human erythropoietin (rhEPO, 3,000 IU/kg) and iron isomaltoside (IIM, 40 mg/kg) on late-phase liver regeneration following PHx and investigated the possible underlying mechanism. Rats administrated with rhEPO showed significantly higher liver mass restoration, interleukin-6 (IL-6, a hepatocyte mitogen) levels, and Ki-67-positive hepatocytes on day 7 after PHx than saline-treated controls. These beneficial effects were further enhanced on days 7 and 14 by co-treatment with IIM. This combination also significantly improved liver function indices, such as increased albumin production and decreased bilirubin levels, but did not alter serum levels of toxic parameters, such as aspartate transaminase and alanine transaminase. This study demonstrates that the combination of rhEPO and IIM synergistically improves late-phase liver regeneration and function after PHx, probably by promoting IL-6-mediated hepatocyte proliferation without adverse effects. Thus, this combination treatment can be a potential therapeutic strategy for patients undergoing resection for hepatic malignancies.

• Asian Pacific Journal of Cancer Prevention
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• 제16권9호
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• pp.3715-3721
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• 2015

Leukemia is a clonal disorder with blocked normal differentiation and cell death of hematopoietic progenitor cells. Traditional modalities with most used radiation and chemotherapy are nonspecific and toxic which cause adverse effects on normal cells. Differentiation inducing therapy forcing malignant cells to undergo terminal differentiation has been proven to be a promising strategy. However, there is still scarce of potent differentiation inducing agents. We show here that Angelica sinensis polysaccharide (ASP), a major active component in Dong quai (Chinese Angelica sinensis), has potential differentiation inducing activity in human chronic erythro-megakaryoblastic leukemia K562 cells. MTT assays and flow cytometric analysis demonstrated that ASP inhibited K562 cell proliferation and arrested the cell cycle at the G0/G1 phase. ASP also triggered K562 cells to undergo erythroid differentiaton as revealed by morphological changes, intensive benzidine staining and hemoglobin colorimetric reaction, as well as increased expression of glycophorin A (GPA) protein. ASP induced redistribution of STAT5 protein from the cytoplasm to the nucleus. Western blotting analysis further identified that ASP markedly sensitized K562 cells to exogenous erythropoietin (EPO) by activating EPO-induced JAK2/STAT5 tyrosine phosphorylation, thus augmenting the EPO-mediated JAK2/STAT5 signaling pathway. On the basis of these findings, we propose that ASP might be developed as a potential candidate for chronic myelogenous leukemia inducing differentiation treatment.

• Journal of Microbiology and Biotechnology
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• 제18권12호
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• pp.1945-1952
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• 2008

Sialylation, the attachment of sialic acid residues to a protein, can affect the biological activity and in vivo circulatory half-life of glycoproteins. Human ${\alpha}2$,3-sialyltransferase (${\alpha}2$,3-ST) and ${\beta}1$,4-galactosyltransferase (${\beta}1$,4-GT) are responsible for terminal sialylation and galactosylation, respectively. Enhanced sialylation of human erythropoietin (EPO) by the expression of ${\alpha}2$,3-ST and ${\beta}1$,4-GT was achieved using recombinant Chinese hamster ovary (CHO) cells (EC1). The sialic acid content and sialylation of N-glycans were evaluated by HPLC. When ${\alpha}2$,3-ST was expressed in CHO cells (EC1-ST2), the sialic acid content (moles of sialic acid/mole of EPO) increased from 6.7 to 7.5. In addition, the amount of trisialylated glycans increased from 17.3% to 26.1 %. When ${\alpha}2$,3-ST and ${\beta}1$,4-GT were coexpressed in CHO cells (EC1-GTST15), the degree of sialylation was greater than that in EC1-ST2 cells. In the case of EC1-GTST15 cells, the sialic acid content increased to 8.2 and the proportion of trisialylated glycans was markedly increased from 17.3% to 35.5%. Interestingly, the amount of asialoglycans decreased only in the case of GTST15 cells (21.4% to 14.2%). These results show that coexpression of ${\alpha}2$,3-ST and ${\beta}1$,4-GT is more effective than the expression of ${\alpha}2$,3-ST alone. Coexpression of ${\alpha}2$,3-ST and ${\beta}1$,4-GT did not affect CHO cell growth and metabolism or EPO production. Thus, coexpression of ${\alpha}2$,3-ST and ${\beta}1$,4-GT may be beneficial for producing therapeutic glycoproteins with enhanced sialylation in CHO cells.

• Biomolecules & Therapeutics
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• 제10권1호
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• pp.59-69
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• 2002

Recently, recombinant human erythropoietin (rHu-EPO) has been used to treat various types of anemia. YHB216 is a new rHu-EPO developed by Yuhan Research Institute. In this study, we investigated the single dose and 4-week repeated dose toxicity of YHB216 in Beagle dogs. In the single dose toxicity study, YHB216 was administered intravenously at single dose levels of 0 and 25,000 IU/kg to dogs (2 dogs/sex/group). There were no treament-related changes in survivals, clinical signs, body weight gain, hematological values, blood chemical values, and necropsy finding during experimental period. In the repeated dose toxicity study, YHB216 was administered intravenously to dogs for 4 weeks at the dose levels of 0, 100, 500, and 2,500IU／kg (3 dogs/sex／group). There were no toxicologically significant changes in clinical signs, body weights, food and water consumptions, ophthalmoscopy, urinalysis and blood chemistry. There were increased values of red blood cell, hemoglobin, and hematocrit at all treated groups. Spleen revealed increased weight and extramedullary hematopoiesis at 500 IU/kg or more. These changes are all considered to be Pharmacology-related effects and were recovered after 4-week recovery period. From these results, it is concluded that LD50 value was above 25,000 IU/kg in the single dose toxicity study of YHB216 in dogs and the no observed adverse effect level (NOAEL) was 100 IU/kg day in the repeated dose toxicity study of YHB216 in dogs.