• The Korean Journal of Nuclear Medicine Technology
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• v.13 no.3
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• pp.143-146
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• 2009

Purpose: We use the centrifugation of refrigeration state in separation of blood serum, Anti-ds-DNA, Vitamin $B_{12}$/Folate and GAD-Ab assay. However, Cortisol urine and 25-Hydroxyvitamin $D_3$ ($25OHD_3$) are conducted centrifuge at room temperature. This is troublesome that change centrifugation temperature into room temperature due to using of most assays at cold temperature. Therefore when using centrifuge after extraction of Cortisol urine and $25OHD_3$, we conducted researches on effect of the centrifugation temperature in assay results. Materials and Methods: In Cortisol urine, add dichloromethane 1.0 mL in urine $500\;{\mu}L$, mix for 15 minutes, and then centrifuge for 8 minutes at 2600rpm. In $25OHD_3$ add acetonitrile 0.5 mL in serum $200\;{\mu}L$, and then centrifuge for 8 minutes at 2600rpm. Those experiments were conducted centrifuge at room temperature and $4^{\circ}C$. And experiments conducted immediately after centrifugation at $4^{\circ}C$ and standing for 20 minutes after centrifugation $4^{\circ}C$. Results: In Cortisol urine, room temperature result in 1.93, 2.18, 2.43, 9.45, 14.2 (${\mu}g/dL$). Experiments of performing immediately after centrifuge at $4^{\circ}C$ result in 1.8, 2.0, 2.3, 8.1, 13.7 (${\mu}g/dL$). Experiments of performing after 20 minutes result in 2.1, 2.1, 2.7, 9.95, 14.35 (${\mu}g/dL$). On the other hand, the $25OHD_3$ tests conducted at room temperature result in 7.13, 26.6, 35.8, 48.2, 74.8 (ng/dL). Experiments were conducted immediately by pipetting after $4^{\circ}C$ centrifugation result in 7.53, 30.9, 40.3, 61.5, 89.1 (ng/dL) as results are higher than experiments at room temperature. The experiments that conducted centrifuge at $4^{\circ}C$ and then left at room temperature for 20 minutes result in 7.40, 32.4, 41.3, 51.6, 85.6 (ng/dL). Conclusions: Experiments were conducted by using centrifuge at $4^{\circ}C$ are higher or lower than room temperature. The differences between results of standing for 20 minutes after centrifuge at $4^{\circ}C$ and those of centrifuge at room temperature are less than conducting immediately. It is concerned that experiments conducted immediately after centrifuge at $4^{\circ}C$ are incorrect, because tubes become dim due to temperature differences between $4^{\circ}C$ and room temperature. Therefore, it is desirable to centrifuge at room temperature as manual and we should pipet promptly without stopping.

• Journal of fish pathology
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• v.20 no.2
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• pp.109-117
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• 2007

This study was performed to identity non hemolytic streptococcus from cultured flounder (Paralichthys olivaceus) with Streptococcosis in the Jeju island. The result of BIOLOGTM test was Streptococcus uberis that simility of 0.5 and 98% identified in MicroLogTM system (Release 4.05). Carbohydrate utility pattern was dextrin, N-acetyl-D-glucosamine, arbutin, maltose, maltotriose, D-cellobiose, D-fructose, D-mannose, α-D-glucose, D-mannitol, β-methyl D-glucoside, salicin, sucrose, D-trehalose, pruvatic acid methyl ester, mono-methyl succinate, glycerol. In addition hemolysis test for S. parauberis and were S. iniae hemolysis in BAP (Blood agar plate). Antibiotic test for S. parauberis were Ampicillin, Amoxicillin and Fluoroquinolone sensitivity. Mutiplex PCR assay were detected S. pauberis (718 bp), S. iniae (870 bp) L. garviae (1,100 bp). Dectected S. parauberis (718 bp) were result of 16S rRNA sequence identified with S. parauberis (Gene bank accession number X89967). All isolated S. parauberis that with bouned by one group. The result were S. pauberis that γ-hemolytic chain form cocci and negative reaction of catalase, Multiplex PCR assay were 718 bp amplicon size.

• THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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• v.3 no.1
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• pp.99-128
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• 1997

In order to investigate the effects of Kyegyoksan on antitumor effects after Sarcoma 180 cells transplantation into the peritoneal cavity or left groin in mice, and immune depression in mice induced by methotrexate, the extracts of its herbal medicines were orally administered for 14 or 21 days. Experimental studies were performed for measureance of $IC_{50}$ in MTT assay, mean survival days, tumor and body weights for antitumor effects, delayed type hypersensitivity, hemagglutinine titer, hemolysin titer, rosette forming cells, interleukin-2 productivity, lymphocyte transformation, natural killer cell activity and phagocytic activity for immune responses in the immune depressed ICR mice, and SGOT, SGPT, BUN and creatinine for liver and kidney protective function in SO-rats. The results were obtained as follows: 1. From the results of MTT assay, the Kyegyoksan exstracts for SUN-1 and SUN-C4 were inhibited cell viability. 2. Mean survival time in Kyegyoksan-treated group was slightly increased with no effectiveness, as compared with the control group. 3. Tumor weight in Kyegyoksan-treated group was depressed with the statistical significance, as compared with the control group(p<0.01). 4. Body weight in Kyegyoksan-treated group was incresed with the statistical significance, as compared with the control group(p<0.05). 5. Delayed type hypersensitivity in Kyegyoksan-treated group was slightly incresed with no effctiveness, as compared with the control group. 6. Hemagglutinin titer in Kyegyoksan-treated group was incresed with the statistical significance(p<0.05), but hemolysin titer was slightly incresed with no effectiveness, as compared with the control group. 7. Rosette forming cells in Kyegyoksan-treated group was incresed with the statistical significance, as compared with the control group(p<0.001). 8. Interleukin-2 productivity in Kyegyoksan-treated group was incresed with the statistical significance, as compared with the control group(p<0.001). 9. Lymphocyte transfomation in Kyegyoksan-treated group was incresed with the statistical significance, as compared with the control group(p<0.01). 10. Natural killer cell activity in Kyegyoksan-treated group at E/T ratio 100 : 1 was incresed with the statistical significance(p<0.01), but at E/T ratio 50 : 1 and 10 : 1 was slightly incresed with no effectiveness, as compared with the control group. 11. Phagocytic activity in Kyegyoksan-treated group was slightly incresed with no effectiveness, as compared with the control group. 12. The levels of serum glutamic oxalacetic transaminase, serum glutamic pyruvic transaminase, blood urea nitrogen and serum creatinine in Kyegyoksan-treated group were not effective change, as compared with the control group. According to the above results, it could be suggested that Kyegyoksan have prominent antitumor effects, enhance both cellular and humoral immunity, and have no injury to liver and kidney functions.

• THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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• v.4 no.1
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• pp.177-197
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• 1998

This experimental study was carried out to evaluate the effect of Yipahnsan on angiogenic inhibition mechanism. This study investigates the effects of Yipahnsan on angiogenic inhibition mechanism evaluate cell adhesive inhibition effect, DNA fragmantaion analysis, nuclear condensation assay, FACScan analysis, angiogenic lumen formation assay, immunocytochemistry analysis, RT-PCR for mRNA expression, western blot analysis, confocal analysis for $Ca^{2+}influx$. The results were summarized as follows : 1. The cell adhesive inhibition ability was strongly increased from $5{\mu}g/ml$ on ECV304 cell line and ECVPAR cell line. 2. YY water extract caused $G_0/G_1$ arrest peak to existed on the ECV304 cell line. 3. YY water extract caused inhibition of proliferation and inducement of apoptosis on the collagen coated plate in ECV304 cell line. 4. YY water extract inhibited the lumen formation on the matrigel coated plate in ECV304 cell line. 5. YY water extract inhibited the expressions of LFA-1 and ELAM-1 on ECV304 cell line and ECVPAR cell line. 6. YY water extract inhibited the expressions of MMP-9 and uPA on ECV304 cell line and ECVPAR cell line. 7. YY water extract inhibited the expression of integrin ${\alpha}_v{\beta}_3$ on ECV304 cell line and ECVPAR cell line. 8. YY water extract decreased the change of $Ca^{2+}$ in intracellular on ECV304 cell line and ECVPAR cell line. According to the results, Yipahnsan showed to be a key antagonist of integrin ${\alpha}_v{\beta}_3$, and to be induction of apoptosis by p53 through flow cytometry. This report also demonstrated that expressions of MMP-9 and uPA were blocked under the angiogenesis model. Thus, we suggested that Yipahnsan blocks angiogenesis by inducing apoptosis in ECV304 and ECVPAR cell lines, and another oriental herbal medicine that treats qi-stagnation and blood-stasis type also has angiogenic inhibition effects.

• Journal of dental hygiene science
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• v.14 no.2
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• pp.223-229
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• 2014

The aim of this study was to examine the effect of polycan-calcium gluconate complex on the levels of immune and inflammatory mediators in gingival crevicular fluid and serum from patients with periodontitis. A total of 39 patients with periodontitis took placebo (placebo group) or polycan-calcium gluconate complex (treatment group) twice a day for 4 weeks. At baseline and 4 weeks, gingival crevicular fluid (GCF) and blood was collected from each subjects. The secretion level of interleukin $(IL)-1{\beta}$ in GCF was measured by enzyme-linked immunosorbent assay (ELISA). Western blot analysis was conducted to determine the expression of matrix metalloproteinase (MMP)-8 and tumor necrosis factor $(TNF)-{\alpha}$ in GCF. Serum samples were analysed for MMP-8 by ELISA and C-reactive protein (CRP) by turbidimetric immuno assay. MMP-8 and $TNF-{\alpha}$ were significantly decreased in the treatment group at 4 weeks. The level of $IL-1{\beta}$ in the treatment group was significantly lower than those of the placebo group. No differences were observed in CRP levels. Taken together, these data indicate that taking of polycan-calcium gluconate complex led to reduction of inflammatory biomarkers in serum and GCF of periodontitis patients.

• Journal of Life Science
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• v.20 no.9
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• pp.1365-1372
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• 2010

We investigated the inhibitory effects of solvent extracts from dried yam on $H_2O_2$-induced oxidative stress and growth of cancer cell lines (HT1080 human fibrosarcoma and HT-29 human colon cancer cells). Yam (Dioscoreacea) has been recognized as a healthy food due to its various biological activities, such as anti-obesity, anti-constipation, anti-proliferation, and anti-mutagenic activities, as well as its ability to decrease blood glucose and cholesterol levels. In order to determine the protective effect on $H_2O_2$-induced oxidative stress, DCFH-DA (dichlorodihydrofluorescin diacetate) assay was conducted. Acetone with methylene chloride (A+M) extract of dried yam appeared to reduce the levels of intracellular reactive oxygen species (ROS) with dose responses. Among the fractions, 85% aq. methanol fraction showed the highest protective effect on production of lipid peroxides. Inhibitory effects of A+M and methanol (MeOH) extracts on the growth of HT1080 and HT-29 cancer cells increased in a dose dependent manner. The treatments of n-hexane, 85% aq. methanol and n-butanol fractions (${\geqq}0.5$ mg/ml concentrations) significantly inhibited the growth of both cancer cells (p<0.05). From these results, 85% aq. methanol fraction showed inhibitory effects on cellular oxidation and growth of human cancer cells, suggesting that this fraction may contain active compounds of dried yam.

• Tuberculosis and Respiratory Diseases
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• v.68 no.5
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• pp.280-285
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• 2010

Background: $QuantiFERON^{(R)}$-TB Gold In Tube (QFT-G IT) has been used for diagnosing latent tuberculosis infection and active tuberculosis (TB) since 2007. However, there has not been enough data on QFT-G IT for universal use in children. In this study, we evaluated the clinical usefulness of the QFT-G IT in pediatric practice. Methods: We retrospectively reviewed the clinical records of 70 patients younger than 18 years of age who had taken QFT-G IT and had a tuberculin skin test (TST) between July 2007 and July 2009 at Wonju Christian Hospital. The subjects were divided into two groups, asymptomatic TB exposure group and disease group. Four patients who were taking immunosuppressants during the study period were excluded. Results: A total of 66 immunocompetent children were included in this study. Among 27 asymptomatic children who had contact histories of TB, 6 (22.2%) were found to be positive by QFT-G IT. Eleven (40.7%) and 5 (18.5%) children were found to be positive by TST with cutoff values of ${\geq}5mm$ and ${\geq}10mm$, respectively. Agreement was fair to good between QFT-G IT and TST (${\kappa}=0.59$: cutoff value ${\geq}5mm$, ${\kappa}=0.7$: cutoff value ${\geq}10mm$). In disease group, 14 patients (35.9%) were diagnosed with active tuberculosis, 8/14 (57.1%) were positive on TST and 9/14 (64.3%) on QFT-G IT. The positive rate of acid-fast bacilli smear, TB-polymerase chain reaction, and culture for tuberculosis was 11% (1/9), 27.3% (3/11) and 33.3% (3/9), respectively. Conclusion: Our data support that the QFT-G IT can be used as an additional diagnostic tool for latent and active tuberculosis infection in children.

• Clinical and Experimental Pediatrics
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• v.55 no.3
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• pp.88-92
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• 2012

Purpose: Mucopolysaccharidosis type II (MPS II or Hunter syndrome) is a rare lysosomal storage disorder caused by iduronate-2-sulfatase (IDS) deficiency. MPS II causes a wide phenotypic spectrum of symptoms ranging from mild to severe. IDS activity, which is measured in leukocyte pellets or fibroblasts, was reported to be related to clinical phenotype by Sukegawa-Hayasaka et al. Measurement of residual plasma IDS activity using a fluorometric assay is simpler than conventional measurements using skin fibroblasts or peripheral blood mononuclear cells. This is the first study to describe the relationship between plasma IDS activity and clinical phenotype of MPS II. Methods: We hypothesized that residual plasma IDS activity is related to clinical phenotype. We classified 43 Hunter syndrome patients as having attenuated or severe disease types based on clinical characteristics, especially intellectual and cognitive status. There were 27 patients with the severe type and 16 with the attenuated type. Plasma IDS activity was measured by a fluorometric enzyme assay using 4-methylumbelliferyl- ${\alpha}$-iduronate 2-sulphate. Results: Plasma IDS activity in patients with the severe type was significantly lower than that in patients with the attenuated type ($p$=0.006). The optimal cut-off value of plasma IDS activity for distinguishing the severe type from the attenuated type was 0.63 $nmol{\cdot}4hr^{-1}{\cdot}mL^{-1}$. This value had 88.2% sensitivity, 65.4% specificity, and an area under receiver-operator characteristics (ROC) curve of 0.768 (ROC curve analysis; $p$=0.003). Conclusion: These results show that the mild phenotype may be related to residual lysosomal enzyme activity.

• Toxicological Research
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• v.26 no.4
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• pp.261-266
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• 2010

In the workplace, the arsenic is used in the semiconductor production and the manufacturing of pigments, glass, pesticides and fungicides. Therefore, workers may be exposed to airborne arsenic during its use in manufacturing. The purpose of this study was to evaluate the potential toxicity of particulate matters (PMs) doped with arsenic (PMs-Arsenic) using a rodent model and to compare the genotoxicity in various concentrations and to examine the role of PMs-Arsenic in the induction of signaling pathway in the lung. Mice were exposed to PMs $124.4{\pm}24.5\;{\mu}g/m^3$ (low concentration), $220.2{\pm}34.5\;{\mu}g/m^3$ (middle concentration), $426.4{\pm}40.3\;{\mu}g/m^3$ (high concentration) doped with arsenic $1.4\;{\mu}g/m^3$ (Low concentration), $2.5\;{\mu}g/m^3$ (middle concentration), $5.7\;{\mu}g/m^3$ (high concentration) for 4 wks (6 h/d, 5 d/wk), respectively in the whole-body inhalation exposure chambers. To determine the level of genotoxicity, Chromosomal aberration (CA) assay in splenic lymphocytes and Supravital micronucleus (SMN) assay were performed. Then, signal pathway in the lung was analyzed. In the genotoxicity experiments, the increases of aberrant cells were concentration-dependent. Also, PMs-arsenic caused peripheral blood micronucleus frequency at high concentration. The inhalation of PMs-Arsenic increased an expression of phosphorylated Akt (p-Akt: protein kinase B) and phpsphorylated mammalian target of rapamycin (p-mTOR) at high concentration group. Taken together, inhaled PMs-Arsenic caused genotoxicity and altered Akt signaling pathway in the lung. Therefore, the inhalation of PMs-Arsenic needs for a careful risk assessment in the workplace.

• Journal of Life Science
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• v.8 no.5
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• pp.479-485
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• 1998

The induction of nephric duct from Xenopus presumptive ectoderm(animal cap) was studied and the high-dose ef-fect of IGF-1 was investigated. Activin A induce various organs from cultured animal cap explants and the effects are time and dose-dependent. On the induction of nephric duct, the combined-dose of activin A and retinoic acid was very efficient method in reference study. In present study, we used IGF-1 as well as activin A as a combined growth factor. The concentration ranges of growth factors were activin A l00ng/ml an IGF-1 0-500ng/m1. Explants were cultured in combined solution for 3days to the normal embryo arrives at st. 43. In general, when the explant was cultured in high concentration(l00ng/ml) of activin A, it was destroyed, however, nephric duct and other tis-sues were differentiated by adding IGF-1. In addition, eye induction by adding IGF-1 500ng/ml to activin A 1- 100 ng/ml solution was studied. The low concentration of activin A(1ng/ml) have blood-like cell inducing effect and the explant was balloon-shaped, however, the high dose combination with IGF-1 extended the range of eye inductive concentration of activin A.