• Title/Summary/Keyword: prolonged culture

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The Cytotoxic and Anti-proliferative Effects of 3-Hydrogenkwadaphnin in K562 and Jurkat Cells Is Reduced by Guanosine

  • Moosavi, Mohammad Amin;Yazdanparast, Razieh;Sanati, Mohammad Hasan
    • BMB Reports
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    • v.38 no.4
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    • pp.391-398
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    • 2005
  • 3-hydrogenwadaphnin (3-HK) is a new daphnane-type diterpene ester isolated from Dendrostellera lessertii with strong anti-tumoral activity in animal models and in cultures. Here, prolonged effects of this new agent on proliferation and viability of several different cancerous cell lines were evaluated. Using [$^3H$]thymidine incorporation, it was found that the drug inhibited cell proliferation and induced G1/S cell cycle arrest in leukemic cells 24 h after a single dose treatment. The cell viability of Jurkat cells was also decreased by almost 10%, 31% and 40% after a single dose treatment (7.5 nM) at 24, 48 and 72 h, respectively. The drug-treated cells were stained with acridine orange/ethidium bromide to document the chromatin condensation and DNA fragmentation. These observations were further confirmed by detection of DNA laddering pattern in the agarose gel electrophoresis of the extracted DNA from the treated cells. Treatment of K562 cells with the drug at 7.5, 15 and 30 nM caused apoptosis in 25%, 45% and 65% of the cells, respectively. Exogenous addition of $25-50\;{\mu}M$ guanosine and/or deoxyguanosine to the cell culture of the drug-treated cells restored DNA synthesis, released cell arrest at G1/S checkpoint and decreased the apoptotic cell death caused by the drug. These observations were not made using adenosine. However, the drug effects on K562 cells were potentiated by hypoxanthine. Based on these observations, perturbation of GTP metabolism is considered as one of the main reasons for apoptotic cell death by 3-HK.

Cultural characteristics and selection of saline tolerant Pleurotus ostreatus at different NaCl concentration medium (NaCl 함량에 따른 내염성 느타리버섯 선발과 재배적 특성)

  • Choi, Jong-In;Chi, Jeong-Hyun;Ha, Tai-Moon;Ju, Young-Cheol
    • Journal of Mushroom
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    • v.3 no.2
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    • pp.65-70
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    • 2005
  • This study was carried out to select oyster mushroom strains tolerant to salinity and characteristic cultivation by different NaCl concentration. Among the 64 strains, Kimjae-10ho, Nongki-2-1ho, Myungyul, Byungneutari-1ho, Bupyungsoyae-1ho, Sambok and Chunchu-2ho resulted in higher mycelial growth and density on PDA medium containing NaCl. The maxium NaCl concentration possible to fruit body formation was 0.5% NaCl in Myungyul, 1.0% NaCl in Kimjae-10ho and Bupyungsoyae-1ho, 1.5% NaCl in Nongki-2-1ho, Byungneutari-1ho, Sambok and Chunchu-2ho, respectively. Increased NaCl concentration on sawdust medium prolonged culture period, while decreased total quantity and come to be short and thin in Length and thickness of stipe. Higher NaCl concentration in the medium decreased the uptake rate of $K_2O$ and CaO, whereas increased the NaCl uptake rate.

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Adoptive Immunotherapy for Small Cell Lung Cancer by Expanded Activated Autologous Lymphocytes: a Retrospective Clinical Analysis

  • Zhang, Guo-Qing;Li, Fang;Sun, Sheng-Jie;Hu, Yi;Wang, Gang;Wang, Yu;Cui, Xiao-Xia;Jiao, Shun-Chang
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.4
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    • pp.1487-1494
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    • 2015
  • Background: To investigate the clinical efficacy of expanded activated autologous lymphocytes (EAAL) in patients with small cell lung cancer (SCLC). Materials and Methods: A total of 32 SCLC patients were selected and randomly divided into EAAL treatment and control groups, 16 cases in each. EAAL were obtained by proliferation of peripheral blood mononuclear cells (PBMCs) of patients followed by phenotype determination. Clinical data of all patients were recorded. Patients of both groups were followed up and the overall survival (OS) were compared retrospectively. Results: After culture and proliferation in vitro, the percentages of $CD3^+$, $CD3^+CD8^+$, $CD45RO^+$, $CD28^+$, $CD29^+$, $CD8^+CD28^+$ and $CD3^+CD16^+/CD56^+$ cells increased markedly (p<0.05). The OS of the EAAL treatment group was longer than that of control group, but the difference was not statistically significant (p=0.060, HR=0.487, 95%CI 0.228~1.037). 1- to 3-year survival rates in EAAL treatment group were longer than those in control group, but there was still no significant difference (p>0.05). COX multivariate regression analysis showed that the number of chemotherapy cycles and the application of EAAL immunotherapy were independent prognostic factors for SCLC patients. The OS in females and chemotherapy${\leq}6$ cycles were obviously prolonged after EAAL immunotherapy. Conclusions: In vitro induction and proliferation of EAAL is easy and biologically safe. Generally, EAAL adoptive immunotherapy can evidently prolong the OS of SCLC patients.

Exosomes from CIITA-Transfected CT26 Cells Enhance Anti-tumor Effects

  • Fan, Wen;Tian, Xing-De;Huang, E.;Zhang, Jia-Jun
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.2
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    • pp.987-991
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    • 2013
  • Aim: To study anti-tumor effects of exosomes from class II transactivator (CIITA) gene transfected CT26 cells. Methods: In this study, we established an MHC class II molecule-expressing murine colon cancer cell line (CT26-CIITA) by transduction of the CIITA gene. Immune effects in vitro and tumor protective results in vivo were tested and monitored. Results: Exosomes from CT26-CIITA cells were found to contain a high level of MHC class II protein. When loaded on dendritic cells (DCs), exosomes from CT26-CIITA cells significantly increased expression of MHC class II molecules, CD86 and CD80, as compared to exosomes from CT26 cells. In vitro assays using co-culture of immunized splenocytes and exosome-loaded DCs demonstrated that CIITA-Exo enhanced splenocyte proliferation and IFN-${\gamma}$ production of CD4+T cells, while inhibiting IL-10 secretion. In addition, compared to exosomes from CT26 cells, CT26-CIITA-derived exosomes induced higher TNF-${\alpha}$ and IL-12 mRNA levels. A mouse tumour preventive model showed that CT26-CIITA derived exosomes significantly inhibited tumour growth in a dose-dependent manner and significantly prolonged the survival time of tumour-bearing mice. Conclusion: Our findings indicate that CT26-CIITA-released exosomes are more efficient to induce anti-tumour immune responses, suggesting a potential role of MHC class II-containing tumour exosomes as cancer vaccine candidates.

Trends of Government Funded Research for Kampo Medicine in Japan and It's Implication (일본에서의 한방의학(漢方醫學)에 대한 국비 지원 연구 동향과 그 함의)

  • Jeung, Chang-Woon;Choi, Chang-Hyuk;Jo, Hee-Geun;Song, Min-Yeong;Baek, Eun-Hye
    • Journal of Korean Medicine Rehabilitation
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    • v.28 no.1
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    • pp.121-131
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    • 2018
  • Objectives We analyzed the trends of government-funded research on Kampo medicine in Japan to provide advanced evidence to R&D support policy for Korean medicine, and to introduce new research fields and trends to the researchers. Methods We reviewed the researches on Kampo medicine through 'research-er.jp' and 'KAKEN' database which contain R&D status in Japan and scientific research funding project issued by the Japan Ministry of Education, Culture, Sports, Science and Technology. Results Since 1976, government-funded research on Kampo medicine has been continuously announced, and now 533 tasks have been completed or are in progress. The average duration of the study is 2.54 years, but it has been prolonged to 3.52 years in recent years. 4~5 million yen was supported per project for laboratory research, and an average of 44,342 thousand yen was supported per project for specialized laboratory research and clinical research. Conclusions Despite the absence of systematically supporting departments, the researches on Kampo medicine in Japan were qualitatively superior since they focused on providing the scientific basis for clinical application. As competition in the world's traditional medicine market becomes more intense, it is necessary to improve the competitiveness of Korean medicine. Therefore, a keen interest in Korean medicine and active support from the government is needed.

Suppressive Effect of Curcuma Zedoaria Roscoe on Pulmonary Metastasis of B16 Melanoma Cells

  • Hwang Jae-Cheol;Kim Mi-Rang;Jung Young-Jae;Lee Young-Ja;Jung Wun-Suk;Seo Un-Kyo
    • The Journal of Korean Medicine
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    • v.26 no.1 s.61
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    • pp.1-10
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    • 2005
  • Objective: We examined the antimetastatic effect of Curcuma zedoaria Roscoe (CZ) on pulmonary metastasis of B 16 cells. Methods: For 6 weeks, Zedoariae Rhizoma made from dried CZ were dissolved in distilled water and administered to mice 2 weeks before they were injected with B]6 melanoma cells. Mice were given CZ at doses of 250 and 500 mg/kg, and were compared for lung weight, survival days, and NO production. Results: Intake of CZ throughout the experiment extended the average survival time. Intake after B16 cell injection slightly prolonged survival time, but intake before B]6 cell injection did not influence life span. We examined the effect of CZ on macrophage function by measuring NO production. After the macrophages were given CZ for 6 weeks, the amount of NO generated by the macrophages stimulated with LPS in culture medium increased. NO generated by the macrophages also served as a cytotoxic factor against B16 melanoma cells. B16 melanoma-conditioned medium reduced NO production by macrophages. However, CZ treatment reversed the reduction in NO production by the conditioned medium significantly. Conclusion : These findings may suggest that macrophage function-modulating activity by CZ appears to underlie its antimetastatic activity, which leads to a decrease in the number of lung metastatic surface nodules and the extension of life span.

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Controlled Release of Nerve Growth Factor from Sandwiched Poly(L-lactide-co-glycolide) Films for the Application in Neural Tissue Engineering

  • Gilson Khang;Jeon, Eun-Kyung;John M. Rhee;Lee, Ilwoo;Lee, Sang-Jin;Lee, Hai-Bang
    • Macromolecular Research
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    • v.11 no.5
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    • pp.334-340
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    • 2003
  • In order to fabricate new sustained delivery device of nerve growth factor (NGF), we developed NGF-loaded biodegradable poly(L-lactide-co-glycolide) (PLGA, the mole ratio of lactide to glycolide 75:25, molecular weight: 83,000 and 43,000 g/mole, respectively) film by novel and simple sandwich solvent casting method for the possibility of the application of neural tissue engineering. PLGA was copolymerized by direct condensation reaction and the molecular weight was controlled by reaction time. Released behavior of NGF from NGF-loaded films was characterized by enzyme linked immunosorbent assay (ELISA) and degradation characteristics were observed by scanning electron microscopy (SEM) and gel permeation chromatography (GPC). The bioactivity of released NGF was identified using a rat pheochromocytoma (PC-12) cell based bioassay. The release of NGF from the NGF-loaded PLGA films was prolonged over 35 days with zero-order rate of 0.5-0.8 ng NGF/day without initial burst and could be controlled by the variations of molecular weight and NGF loading amount. After 7 days NGF released in phosphate buffered saline and PC-12 cell cultured on the NGF-loaded PLGA film for 3 days. The released NGF stimulated neurite sprouting in cultured PC-12 cells, that is to say, the remained NGF in the NGF/PLGA film at 37 $^{\circ}C$ for 7 days was still bioactive. This study suggested that NGF-loaded PLGA sandwich film is released the desired period in delivery system and useful neuronal growth culture as nerve contact guidance tube for the application of neural tissue engineering.

Methanol Extract of Cassia mimosoides var. nomame and Its Ethyl Acetate Fraction Attenuate Brain Damage by Inhibition of Apoptosis in a Rat Model of Ischemia-Reperfusion

  • Kim, Ki-Hong;Lee, Jong-Won
    • Preventive Nutrition and Food Science
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    • v.15 no.4
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    • pp.255-261
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    • 2010
  • Ischemic stroke, a major cause of death and disability worldwide, is caused by occlusion of cerebral arteries that, coupled with or without reperfusion, results in prolonged ischemia (hypoxia and hypoglycemia) and, ultimately, brain damage. In this study, we examined whether methanol extract of the whole plant of Cassia mimosoides var. nomame Makino that grows naturally in Korea, as well as Japan and China, and some of its fractions obtained by partitioning with organic solvents could protect human hepatocellular carcinoma cells (HepG2) under hypoxic condition by inhibiting apoptosis. We also investigated if these extracts could attenuate brain damage in a rat model of 2 hr of ischemia, generated by middle cerebral artery occlusion, and 22 hr of reperfusion. The whole extract ($100{\mu}g$/mL) maintained the cell number at more than half of that initially plated, even after 24 hr of cell culture under hypoxic condition (3% $O_2$). In the absence of the whole extract, almost all of the cells were dead by this time point. This improvement of cell viability came from a delay of apoptosis, which was confirmed by observing the timing of the formation of a DNA ladder when assessed by gel electrophoresis. Of fractions soluble in hexane, ethyl acetate (EA), butanol and water, EA extracts were selected for the animal experiments, as they improved cell viability at the lowest concentration ($10{\mu}g$/mL). The whole extract (200 mg/kg) and EA extract (10 and 20 mg/kg) significantly reduced infarct size, a measure of brain damage, by 34.7, 33.8 and 45.2.0%, respectively, when assessed by 2,3,5-triphenyl tetrazolium chloride staining. The results suggest that intake of Cassia mimosoides var. nomame Makino might be beneficial for preventing ischemic stroke through inhibition of brain cell apoptosis.

Influence of Substrates on the Isozyme Patterns of Cellulase and Xylanase Complexes in Aspergillus niger (Aspergillus niger에 있어서 섬유질 분해효소계의 동질효소 양상에 미치는 기질의 영향)

  • Rho, Jae-Rang;Rhee, Young-Ha;Chung, Jae-Hoon
    • The Korean Journal of Mycology
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    • v.18 no.4
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    • pp.209-217
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    • 1990
  • The influence of cellulosic and hemicellulosic substrates on the production of cellulase and xylanase complexes in Aspergillus niger was investigated. The culture conditions with different substrates exhibited profound effects on the level of endoglucanase (CMCase), ${\beta}-glucosidase$, endoxylanase and ${\beta}-xylosidase$, and on their isozyme patterns. However, intracellular and extracellular isozyme patterns of cellulase and xylanase complexes were qualitatively identical and appeared to be simultaneous in the early growth phase. Prolonged incubation led to the increase in the concentrations of isozymes with a little changes in the relative proportions of those isozymes. These results suggest that the biosynthesis of cellulase and xylanase complexes in A. niger is coordinately regulated at the level of induction. Moreover, multiple forms of extracellular cellulase and xylanase complexes seem to be the outcome of specific gene expression and should not be considered solely as the consequence of post-secretional modification of synthesized enzymes.

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Influence of Job Stress and Resilience on the Burnout of Nurses Who Works at the Designated Public Relief Hospital (국민안심병원에서 근무하는 간호사들의 직무스트레스와 회복탄력성이 소진에 미치는 영향)

  • Shin, Rok;Baek, Hye-Jin;Ahn, Da-Bin
    • The Journal of the Korea Contents Association
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    • v.21 no.9
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    • pp.595-608
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    • 2021
  • The purpose of this study is to examine the effects of job stress and resilience on burnout of nurses who take care of patients with COVID-19 in prolonged pandemics. Structured questionnaires were distributed and analyzed for 162 nurses. Inadequate compensation of job stress, job demands, workplace culture, philosophical patterns of resilience, and relational patterns were statistically significant. As a result of determining the effect of job stress and resilience on burnout through the Stepwise multiple regression analysis, the final significant variable was job stress. The burnout of nurses can affect the quality of nursing services, reduced hospital competitiveness, and even quarantine systems. Therefore, this study suggests that proper compensation policy support, such as treatment of nurses and improvement of working conditions, is needed.