• Reproductive and Developmental Biology
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• v.30 no.2
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• pp.81-85
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• 2006

This study was conducted to examine the effect of IRES controlled reporter gene on screening and production of recombinant human erythropoietin (EPO) proteins from cultured CHO cells. The cDNA was cloned for EPO from human liver cDNA Using site-directed mutagenesis, we generated recombinant human EPO (rhEPO) with two additional N-glycosylations (Novel erythropoiesis-stimulating protein: NESP). Wild type hEPO and NESP were cloned into expression vectors with GFP reporter gene under regulatory control of CMV promoter and IRES so that the vectors could express both rhEPO and GFP. The expression vectors were transfected to cultured CHO-K1 cells. Under microscopy, expression of GFP was visible. Using supernatant of the culture, ELISA assay, immunocytochemistry and in vitro assay using EPO dependant cell line were performed to estimate biological activity to compare the production characteristics (secretion levels, etc.) between rhEPO and NESP. The activity of NESP protein, obtained by mutagenesis, was described and compared with its rhEPO counterpart produced under same conditions. Although NESP had less secretion level in CHO cell line, the biological activity of NESP was greater than that of rhEPO. These results are consistent with previous researches. We also demonstrated that rhEPO and GFP proteins expressed simultaneously from transfected CHO cell line. Therefore we conclude that use of GFP reporter gene under IRES control could be used to screen and produce rhEPO in cultured CHO cells.

• Microbiology and Biotechnology Letters
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• v.32 no.2
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• pp.142-148
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• 2004

Erythropoietin is a main regulator of human erythropoiesis. Recombinant human erythropoietin (rhEPO) is one of the glycoproteins produced in animal cells, and it has oligo saccharides chains which comprise about 40％ of its molecular mass. Because the content of sialic acid can extend circulatory lifetime, the high degree of sialylation is often a desirable feature of therapeutic glycoproteins. In this study, the sialylation of rhEPO produced by chinese hamster ovary cell culture was maximized by supplementing the culture medium with N-acetylm-annosamine (ManNAc), a direct intracellular precursor for sialic acid synthesis and 2-deoxy-2,3-dehydro-N-acetylneuraminic acid (NeuAc2en), a sialidase inhibitor. Feeding of 20 mM ManNAc/0.5 mM NeuAc2en into culture medium increased the sialic acid content by nearly tenfold compared with unsupplemented medium. This effect was achieved without affecting the cell growth or product yield. Six erythropoietin fractions differing in sialic acid content, ranging from 11∼15％ of EPO, were identified from chinese hamster ovary cell-derived rhEPO by mono Q column chromatography. It was found that, at 20 mM ManNAc/0.5 mM NeuAc2en feeding, productivity of hyper-glycosylated EPO increased up to 50％, compared with the unsupplemented medium.

• Applied Microscopy
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• v.33 no.1
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• pp.41-48
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• 2003

The hemopoiesis in human fetal spleen was studied with transmission electron microscope. There were undifferentiated proerythroblast, basophilic erythroblast, polychromatophilic erythroblast, and acidophilic erythroblast. Besides, enucleated nuclei and mitoses were present. Groups of erythroblastic cells were surrounded by certain cell. The structure was identical to erythropoietic island found in fetal liver. So, erythropoisis in spleen was developing in a pattern similar to fetal liver. Megakaryobalst were found in spleen, but there was no mature cells, cells in mitosis nor platelet formation. It was not clear whether megakaryoblast in circulation was trapped in spleen or participated in megakaryopoiesis. In summary, erythropoiesis took place in fetal spleen in a pattern similar to fetal liver and bone marrow. But it was not certain whether megakaryopoiesis took place in fetal spleen.

• Korean Journal of Community Nutrition
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• v.5 no.3
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• pp.493-501
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• 2000

The purpose of this study was to assess the nutritional status of iron of the elderly residing in the Jeonju area. The anthropometric parameters, nutrient intake and biochemical status of iron, were measured for 60 korean elderly(23 elderly men and 37 elderly women aged 60 - 79 years old). The level of hemoglobin(Hb), hematocrint(Hct), serum iron(Fe), total iron binding capacity(TIBC) and serum ferritin(Ferritin) were measured and transferrin saturation(TFsaturation) was calculated. Mean values of Hb, Hct, Fe, TIBC, TFsaturation and Ferritin were 14.49$\pm$0.93g/이, 42.47$\pm$2.59%, 125.48$\pm$52,.46$\mu\textrm{g}$/dl, 338.13$\pm$45.92$\mu\textrm{g}$/dl, 193.00$\pm$125.03$\mu\textrm{g}$/l in elderly men and 12.82$\pm$0.99$\mu\textrm{g}$/dl, 37.66$\pm$2.90%, 100.08$\pm$42.82$\mu\textrm{g}$/dl, 356.41$\pm$54.65$\mu\textrm{g}$/dl, and 99.35$\pm$117.22$\mu\textrm{g}$/l in elderly women, respectively. Prevalence of iron deficiency varied greatly with biochemical indices of iron. It was 13.0% when judged by Fe(60 $\mu\textrm{g}$/dl) and TFsaturation(15%) whereas 34.78% by Hct(41%) in elderly men. However 13.5% of the elderly women showed iron depletion(Ferritin<20$\mu\textrm{g}$/l) and 43.2% showed suppressed erythropoiesis with iron deficiency(TIBC>360$\mu\textrm{g}$/dl). The anemic subjects assessed with TFsaturation(<15%) represented 13.5% of the elderly women, whereas 18.9% of the subjects possessed less than 12g/dl of Hb. The Hb concentration was positively correlated with Hct(r=0.980, p<0.001), Fe(r=0.384, p<0.01) and TFsaturation(r=0.349, p<0.01). On the other hand, Ferritin concentration showed a significantly negative correlation with TIBC(r=0.349, p<0.05) and a positive correlation with TFsaturation(r=0.362, p<0.01). Major food groups of iron intake in the elderly were vegetables, cereals, and fish. The mean daily intake of iron was not significantly different between elderly men and women(12.82mg vs 10.35mg). Intake of heme iron however, was significantly higher(p<0.01) in elderly men(1.03mg) than women(0.42mg). Total absorbable iron caculated by the method of Monsen was 0.55mg, 0.40mg in elderly men and women, respectively and bioavailability of dietary iron 4.29% and 3.87%.

• Tuberculosis and Respiratory Diseases
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• v.42 no.5
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• pp.654-659
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• 1995

Background: Tuberculosis has commonly been associated with various hematologic changes. A difference between the changes found in pulmonary tuberculosis and those found in miliary tuberculosis has been discussed. Up to now some worker were investigated hematological changes associated with pulmonary tuberculosis but was not investigated those associated with miliary tuberculosis in korea. Therefore we examimed the peripheral blood and bone marrow findings in miliary tuberculosis patients to determine hematologic changes. Methods: We performed blood sample at admission and bone marrow biopsy within 7days after admission. For evaluation of the hematologic findings, full blood counts and marrow differential counts were defined by the criteria outlined by Dacie and Lewis. Results: 1) Peripheral blood findings: Pancytopenia in 10% of patients, anemia in all patients, leukocytosis in 10% of patients, leukopenia in 20% of patients, thrombocytopenia in 30% of patients, lymphocytopenia in 90% of patients, monocytosis in 40% of patients and neutrophilia in 10% of patients were found at peripheral blood. 2) Bone marrow findings: Lymphocytopenia in 30% of patients, lymphocytosis in 20% of patients, plasmacytosis in 40% of patients, monocytosis in 100% of patients, and hypocellularity in 30% of patients were found at bone marrow. Erythropoiesis was decreased in 30% of patients. Granulopoiesis was decreased in 20% of patients and increased in 20% of patients. Bone morrow granuloma occured in 25% of patients. Conclusion: Hematologic changes of miliary tuberculosis were seen tendency of cytopenic pattern but monocyte was increased at peripheral blood and bone morrow. This findings would provide additional information for the differential diagnosis of miliary tuberculosis.

• Clinical and Experimental Pediatrics
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• v.52 no.4
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• pp.435-440
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• 2009

Purpose : The concentration of soluble transferrin receptor (sTfR) is estimated as an iron parameter to evaluate erythropoiesis and iron status. The aim of our study is to evaluate the correlation between sTfR concentration and inflammatory parameters and to distinguish iron deficiency anemia from anemia of inflammation. Methods : One hundred and forty-four infants younger than two years of age who visited Gyeongsang University Hospital for 7 years from 2000 to 2006 were enrolled. Patients who had hemoglobin (Hb) <11 g/dL and ferritin <12 mg/L were excluded. Routine hematologic lab, serum ferritin, sTfR, and inflammatory markers [C-reactive protein(CRP), interleukin-6(IL-6), and absolute neutrophil count (ANC)] were investigated. Results : In all patients, the sTfR concentration showed a correlation with Hb, ferritin, MCV, and ANC, but not with CRP and IL-6. In multiple regression models, positive correlations were found between sTfR concentration and IL-6 (r=0.078, P=0.043), and negative correlations were found between sTfR concentration and ANC (r=-0.117, P=0.033) and MCV (r=-0.027, P=0.009). Conclusion : sTfR concentration was influenced by inflammatory parameters. Therefore, sTfR does not appear to be a useful parameter for discriminating between iron deficiency anemia and anemia of inflammation in infants.

• Clinical and Experimental Pediatrics
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• v.52 no.2
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• pp.167-175
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• 2009

Purpose : This study has been conducted to analyze whether the biochemical nutrition indexes might be useful and effective for evaluating the nutrition states of children. Methods : We evaluated 269 children, aged 3-9 years old, who had visited Gangneung Asan Hospital for elective surgery from January 2006 to December 2007, and examined their anthropometric and preoperative laboratory data with retrospective analysis. The children were classified into underweight, normal weight, overweight, and obese groups according to body mass index (BMI). The biochemical nutrition indexes (total lymphocyte count (TLC), hemoglobin, hematocrit, serum albumin, cholesterol, et al) of each group were then analyzed statistically. Results : None of the groups showed statistically significant differences in TLC. Serum albumin decreased significantly in the underweight group. Red blood cell (RBC) count, hemoglobin, hematocrit, and serum total cholesterol in the obese group were higher than in the normal weight group. None of the groups showed statistically significant increase in mean corpuscular volume or mean corpuscular hemoglobin, and it seems that the increase of hemoglobin and RBC count in the overweight and obese groups is due to the enhancement of erythropoiesis rather than iron metabolism. However, in females, almost all nutrition indexes except albumin were statistically significantly poor. Conclusion : Serum albumin, total cholesterol, RBC count, hemoglobin, and hematocrit were useful as nutrition indexes. However, except for albumin, these indexes were significantly poor for females. More control studies are needed to confirm the effectiveness of biochemical indexes for evaluating the nutritional state of children.

• Tuberculosis and Respiratory Diseases
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• v.66 no.2
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• pp.104-109
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• 2009

Background: Anemia is quite common in lung cancer patients and known to decrease the quality of life. Darbepoetin alfa is an erythropoiesis-stimulating protein approved for administration to cancer patients. This study examined the efficacy and safety of darbepoetin alfa in lung cancer patients with a hemoglobin concentration <10 g/dl during chemotherapy. Methods: Lung cancer patients (n=178) received darbepoetin alfa at doses of 1.91 ${\mu}g/kg$ per week until the hemoglobin concentration increased to >10 g/dl. The efficacy and safety were measured by comparing the hemoglobin concentration and assessing the adverse events. Results: After chemotherapy, the hemoglobin concentration decreased to 9.03${\pm}$0.64 g/dl. With the darbepoetin alfa treatment, the hemoglobin concentration increased to 10.09${\pm}$1.17 g/dl after 4 weeks reaching a peak hemoglobin concentration of 10.45${\pm}$1.18 g/dl. The changes in hemoglobin after 4 and 8 weeks with treatment were 1.08${\pm}$1.24 g/dl and 1.38${\pm}$1.59 g/dl (p<0.01). At least a 1 g/dl or more increase in hemoglobin was observed in 62.4% of patients. There were no serious adverse effects except for some mild reactions. Conclusion: Darbepoetin alfa administered to lung cancer patients appears to be an effective, well-tolerated treatment for chemotherapy induced anemia.

• Journal of Nutrition and Health
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• v.43 no.2
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• pp.114-122
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• 2010

Se and Fe are trace minerals acting as antioxidant scavenging free radicals. Iron deficiency is the most frequently reported nutritional deficiency in females. Body iron status are known to be dependent not only upon dietary iron intake, but also upon micro-mineral nutrition and obesity. Antioxidants such as selenium are reported to play an important role on the regulation of erythropoiesis by protecting RBC membrane from antioxidative damage. In this study, iron status in young females and its relationships with selenium status and physique were examined. Serum selenium and iron concentrations were measured by HANARO research reactor using neutron activation analysis method (NAA-method). The proportion with iron deficiency and anemia were 27.1% and 8.6%, respectively in young females, but the proportion with iron deficient anemia was 1.4%. The mean serum selenium level was $12.0\;{\mu}g/dL$ and in normal range in the young women. The study participants were tertiled according to BMI and serum selenium levels. Serum ferritin and iron levels inclined with increasing BMI tertiles. Serum iron and RBC count were higher in middle selenium group than low selenium group. Individuals had significantly lower hematocrit level in the lowest tertile for their serum selenium levels compared with the highest tertile. The serum ferritin level was predicted 25% by BMI and RBC count 26.2% by the serum selenium level and body fat%. In conclusion, this study shows that body iron status in young adult females are influenced by obesity and body selenium status.

• 한국체육학회지인문사회과학편
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• v.54 no.1
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• pp.553-566
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• 2015

This study was performed to identify the effects of different intensities of regular aerobic exercise on erythropoietin (EPO) and BDNF levels, and cognitive function and working memory in middle-aged women. Women aged 40 to 60 years residing in G-gu, Y-si, Gyeonggi-do were divided into 3 groups: control group, moderate-intensity aerobic exercise group and high-intensity aerobic exercise. All groups were asked to exercise at the given intensities, twice a week for a total of 12 weeks. Blood samples were collected from participants on week 0 (before exercising), week 6 and week 12, and then cognitive function and working memory tests were followed to measure erythropoietin (EPO) and BDNF levels, cognitive function and working memory. Repeated measures ANOVA, univariate analysis and follow-up test were performed on all data to compare the group, period and interaction through a SPSS. As a result, a significant difference over time was observed in EPO, BDNF, cognitive function and working memory; therefore, a follow-up one-way ANOVA analysis was performed on each group. As a result of analysis, a significant increase in erythrocyte, hematocrit, BDNF level and working memory was observed in moderate-intensity aerobic exercise group while erythrocyte and working memory were significantly increased inhigh-intensity aerobic exercise group. When comparing the results between the groups, the level of hematocrit was shown to be significantly higher in both moderate-and high-intensity aerobic group than the control group and also the higher level of hemoglobin was observed in both moderate-and high-intensity aerobic group comparing to control group. Considering the results of this study, therefore, a 12-week long aerobic exercise at moderate to high intensity positively affected EPO and BDNF levels, cognitive function and working memory in middle-aged women.