• KSBB Journal
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• v.28 no.2
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• pp.86-91
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• 2013

To date, various strategies have been studied to increase specific productivity in Chinese hamster ovary (CHO) cell cultures. Also, albumin-fusion platform is being applied to other important bioactive peptides with short half-lives. Here, we investigated the effects of silkworm gland hydrolysate (SGH) on the production of albumin-erythropoietin (Alb-EPO) in transgenic CHO cells. The viable cell density of CHO cells was increased by 13% in the medium containing 1 mg/mL SGH higher than in the control medium without SGH. In addition, the production of Alb-EPO was also 1.26- fold enhanced by reducing the early apoptosis of CHO cells. In conclusion, SGH could be used as a useful supplement for the enhancement of recombinant protein production.

• KSBB Journal
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• v.31 no.4
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• pp.270-276
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• 2016

In glycoprotein, Terminal sialic acid residues of N-linked glycan are imperative things because they prevent the recognition from asialoglycoprotein-receptor that affect the half-life of glycoproteins. So establishment of culture process for enhancing sialic acid is important to maximize sialic acid contents of glycoprotein. In this study, we investigated effects of biphasic culture of Chinese hamster ovary (CHO) cells producing albumin-erythropoietin to increase sialylation. Biphasic cultures were performed with shift of $CO_2$ concentrations and temperatures at day 5 when viable cell density was decreased and sialidase was started to be released by cell lysis. The examined temperature set points were 33, 35 and $37^{\circ}C$ respectively and the $CO_2$ concentration was 1, 5, 10 and 15%. We confirmed that sialidase activity was the lowest in biphasic culture that was shifted from normal culture condition to 1% of $CO_2$ and $33^{\circ}C$ on day 5. However, the temperature and concentration of $CO_2$ have little effect on activity of ${\alpha}2,3$-sialyltransferase. Also, sialic acid contents were enhanced 1.13-fold higher than that in control culture. In conclusion, Biphasic cultivation in CHO cells led to inhibition of sialidase activity and increases of sialylated glycan.

• KSBB Journal
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• v.30 no.1
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• pp.44-51
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• 2015

Chinese hamster ovary (CHO) cells are the most widely used mammalian host for the commercial production of recombinant proteins. However, they show relatively low yields of recombinant proteins in comparison with microbial cells. Various strategies have been tried to overcome this drawback. The acetyl moieties are attached to the N-terminus of histone by histone acetyltransferase (HAT) while histone deacetylase (HDAC) removes histone-bound acetyl groups. HDAC inhibitor (HDACi), such as sodium butyrate, sodium propionate and valproic acid, can enhance specific productivity of CHO cells. Human albumin-erythropoietin (Alb-EPO) is a novel 105 kDa protein comprising recombinant human EPO fused to human albumin. In this study, we examined the effects of HDACi on the production of Alb-EPO in CHO cells with various concentrations in the range of 0-1 mM. The results showed that sodium butyrate was found to be the best HDACi for enhancing productivity. It enhanced not only the production of Alb-EPO but also the apoptosis of recombinant CHO cells.

• KSBB Journal
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• v.13 no.3
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• pp.295-302
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• 1998

Human Erythropoietin (EPO) gene is cloned in quail fibrosarcoma cell, QT35. Because molecular weight of EPO is similar to that of serum albumin, cell culture with serum containing medium makes purification of EPO very difficult. Using fractional factorial study, we have developed serum free medium for the recombinant QT35 cell lines, QT N4D4 and QT SY-IMP, which have cloned EPO with glutamine synthetase (GS) gene amplification system and with puromycin selective marker, respectively. Among the seven frequently used medium components, fibronectin, BSA, and EGF were the most important for EPO production. However, sufficient fibronectin supplement to the medium did not make any good attachment of QT35 to culture plate over 3 days. Therefore, to maximize EPO production, we attempted a medium-shift at confluence from serum containing medium to serum free medium(QT SFM6). Using the medium-shift protocol with QT SFM6, nearly the same productivity of EPO was achieved comparing with that without medium-shift. This result was true in both QT35 cell lines in three types of culture, i.e. T flask, microcarrier and roller bottle cultures.

• BMB Reports
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• v.53 no.3
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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.

• Korean Journal of Clinical Pharmacy
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• v.21 no.2
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• pp.122-130
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• 2011

Objective: Although recombinant human erythropoietin (rhEPO) has revolutionized the treatment of anemia in chronic kidney disease (CKD) receiving hemodialysis (HD) with no need of blood transfusion, some patients have a blunted or appear to be resistant to rhEPO. There is a controversy in the causes of rhEPO resistance in maintenance HD patients with anemia. This study is to examine current anemia treatment outcomes and the factors influencing the rhEPO responsiveness in HD patient with CKD. Methods: The clinical parameters or factors relating to erythrompoietin treatment outcomes and erythropoietin responsiveness were collected from the HD patients in two large dialysis centers for three months. The collected paramenters included serum iron, total iron biding capacity (TIBC), transferrin saturation rate, ferritin, albumin, intact PTH, C-reactive protein (CRP), nPCR and medications such as an angiotensin converting enzyme inhbitor, an angiotension II receptor blocker and an HMG-CoA reductase inhibitor (HMG-CoA RI). The data were analyzed to examine the degree of acheiveing the anemia treatment goal and factors relating to ERI. Results: Among total 111 patients, 42 (42.3%) and 47 (37.8%) patients achieved the target Hct and Hb based on the Health Insurance Review and Assessment Services (HIRA) reimbursement criteria. In the higher ERI group (upper quartile), the patients had higher CRP levels (0.5 mg/dl) (p=0.0096), and lower TIBC score (<$240{\mu}g/dl$) (p=0.0027), and less patients were taking HMG-CoA RI (p=0.0019). Male patients (p=0.0204), patients with high TIBC score ($R^2$=0.084, p=0.0021) and patients taking HMG-CoA RI (p=0.0052) required to administer less dose of rhEPO meaning higher erythropoietin responsiveness. Conclusion: Less than 50% of CKD patients were achieving the goals of anemia by erythropoietin administration in large hospitals in Korea even though the goals were lower than those of NKF-K/DOQI practice guideline. The factors influencing ERI were sex, TIBC and HMG-CoA RI administration status, and neither an ACEI nor an ARB did not influence ERI.

• Journal of Preventive Medicine and Public Health
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• v.43 no.3
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• pp.245-256
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• 2010

Objectives: This study aims to compare quality indicators for the hemodialysis services between patients with health insurance and those with medical aid. Methods: This study used data from sampled hospitals that provided a hemodialysis service. A total of 2287 patients were selected, and the information for hemodialysis service has been granted from medical record reviews. A multi-level regression analysis was used to examine the differences in process and outcome indicators for hemodialysis between patients with health insurance and those with medical aid. Process indicators were defined as: frequency of hemodialysis, hemodialysis time, erythropoietin (EPO) use, measurement of hemodialysis dose at least once a month, measurement of phosphate at least once every three months, and measurement of albumin at least once every three months. Outcome indicators were defined as: hemodialysis adequacy, anemia management, blood pressure management, and calcium, phosphate and nutrition management. The total scores for outcome indicators ranged from 0 (worst) to 4 (best). Results: There was a significant difference in the measurement of hemodialysis dose at least once a month between patients with health insurance and those with medical aid (OR 0.66, 95% CI = 0.43 - 0.99). However, frequency of hemodialysis, hemodialysis time, EPO use, measurement of phosphate at least once every three months, measurement of albumin at least once every three months, hemodialysis adequacy management, Hb${\geq}$11 g/dL, blood pressure within the range of 100-140 /60-90 mmHg, calcium x phosphate${\leq}$55 $g^2/dL^2$ and albumin${\geq}$4 g/dL were not significantly different between the groups. Conclusions: There were no significant differences in outcome indicators for hemodialysis between the groups. Further studies are warranted into the mechanism that results in no differences in the outcome indicators for hemodialysis.

• Journal of Korean Academy of Fundamentals of Nursing
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• v.16 no.2
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• pp.190-199
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• 2009

Purpose: This study was conducted to identify the factors contributing to sleep disorders in patients on hemodialysis. Methods: A descriptive correlational study design was used. The participants were 135 patients on hemodialysis in hemodialysis clinics. Data were collected from March to May 2007 using structured questionnaires and hematologic sample. Stepwise multiple regression was used to identify factors influencing sleep disorders among the demographic-clinical factors, depression and fatigue. Results: About 69% of the patients had a sleep disorders. There were significant differences in sleep disorders according to marital status, caregiver, religion, economic level, insurance, erythropoietin, somnifacient, and antihypertensive agents. But there were no differences according to age, gender, dialysis period, or antidepressants. Their sleep disorders had significant correlations with depression, and fatigue, and a significant negative correlation with Protein, Albumin, Phosphate and BUN. But there were no correlations with hematocrit, hemoglobin, creatinine, sodium, potassium, or calcium. Depression and fatigue were factors influencing sleep disorders. They accounted for 43.8% of the variance in sleep disorders in these patients. Conclusion: Findings provide an understanding of sleep disorders and the factors that are an influence in patients on hemodialysis. To promote sleep in these patients, nursing interventions to manage depression and fatigue are needed.

• Korean Journal of Adult Nursing
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• v.13 no.3
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• pp.451-462
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• 2001

This study was conducted to examine and compare the characteristics in the cognitive functions of peritoneal dialysis, hemodialysis and normal kidney function groups as basic data for effectively educating dialysis patients. The data were collected from May 10 to October 30. 2000, collected from each of 20 patients with peritoneal dialysis, hemodialysis and normal Kidney function and who registered for the dialysis room at a general hospital affiliated to a university in Seoul and sampled by age and educational level through personal interviews with the researchers of this study. As a measuring tool, MMSE developed by Folstein et al.(1975) to measure cognitive function disorder was used, slightly revised for hemodialysis patients. Collected data were processed into frequency, percentage, mean, and standard deviation by the use of SAS. The results of this study are as follows : 1. With a maximum of 30 points for cognitive function, the mean of the peritoneal dialysis patients was $27.06{\pm}2.06$, while that of the hemodialysis patients was $27.25{\pm}2.76$; that of the normal Kidney function patients was $27.85{\pm}2.00$, indicating no significant difference among those three groups. 2. As for the subjects who scored 23 points, the turning point of confirming the cognitive disorder, the percentage was 5% in the case of the peritoneal dialysis, 10% in the case of the hemodialysis and 5% in the case of the normal Kidney function group. 3. Differences between the peritoneal dialysis and hemodialysis patients by gender, occupation, spouse, diabetes, hypertension, the period of dialysis, number of hospitalizations, and the use of erythropoietin were not significant in the scoring of cognitive function. 4. There was no significant correlation between the level of Hb, Hct, albumin, aluminium, PTH, BUN, Cr, dialysis adequacy and the cognitive function. Considering such results, it is clear that there is no significant difference in the cognitive functions of the sampled subjects. Therefore, the nurse in the dialysis room should continually carry out assessment and intervention against elements degrading the effect of patients' education to improve self- care.

• KSBB Journal
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• v.31 no.1
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• pp.66-72
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• 2016

Chinese hamster ovary (CHO) cells have been widely used for production of various recombinant proteins such as cytokines and monoclonal antibodies. The cell aggregation and cell death in CHO cell culture directly affect cell viability, and productivity and quality of products. In this study, we investigated preventing effects of storage-protein 2 (SP2) derived from silkworm hemolymph on cell aggregation and cell death in CHO cell culture producing albuminerythropoietin (Alb-EPO). The viable cell density in the culture supplemented with 2 mg/mL SP2 was 1.71-fold higher than that in control culture. Increased titer of Alb-EPO was also found in the culture with SP2. Morphology of CHO cells in SP2 supplemented cultures did not differ from that of control. In addition, the cell aggregation rate of the SP2 cultures was reduced 20% compared to the control. Finally, we confirmed that the apoptosis was strongly suppressed by addition of SP2 in the cultures. These results clearly demonstrate that SP2 can be served as an effective supplement for enhancing titer of Alb-EPO via reducing cell aggregation and cell death.