• Proceedings of the Korean Vacuum Society Conference
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• 2013.02a
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• pp.599-599
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• 2013

최근의 원자간력현미경(AFM)은 생체물질을 대상으로 여러 구조적 형상뿐만 아니라 물리적 특성 측정이 가능하여 바이오분야에 다양이 활용되고 있다. 줄기세포의 신경세포로 분화 인지에 대한 연구와 관련하여 본 연구에서는 AFM의 한 기능인 Force-Distance curve 측정법을 활용하여 신경암세포주라 불리는 SH-SY5Y를 대상으로 분화 전과 후의 세포막의 stiffness 변화를 측정하였다. 세포막의 stiffness값은 시료표면과 맞닿은 AFM 탐침에 계속적으로 수직방향의 힘이 가해질 시 AFM 캔티레버의 구부러짐 정도로 측정된다. SH-SY5Y는 RA (retinoic acid) 처리에 의해 분화유도 되었으며, 생물학적 방법인 western blotting법을 통해 분화여부를 확인하였다. 측정영역은 AFM topography 이미지 상에서 roughness가 가장 낮은 분화 전과 후 SH-SY5Y의 핵 주변영역으로 선정하였다. 선정된 영역 내에 여러 부분의 분화 전후 세포막의 stiffness 값을 측정하여 통계화한 결과, 분화 전과 후 세포막의 stiffness 차이를 확인할 수 있었다. 분화 전 SH-SY5Y 세포막의 stiffness는 0.79445 N/m인 반면, 분화 후 SH-SY5Y 세포막의stiffness는 0.60324 N/m로 확인되었다. 이는 분화 전에 비하여 분화 후 SH-SY5Y 세포막의 stiffness가 약 24.07% 감소된 것으로 판단할 수 있다. 본 연구는 생물학적 복잡한 방법이 아닌 간단한 방법으로 세포의 stiffness의 변화 측정을 통한 세포의 분화를 판별할 수 있는 방법을 개발한 것으로 여러 줄기세포의 특정세포로 분화여부 판단에 활용할 수 있을 것으로 사료된다.

• Clinical and Experimental Pediatrics
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• v.61 no.2
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• pp.53-58
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• 2018

Purpose: Although the prognosis is generally good in patients with intermediate-risk neuroblastoma, no consensus has been reached on the ideal treatment regimen. This study analyzed treatment outcomes and toxicities in patients younger than 18 months with stage 4 MYCN nonamplified neuroblastoma. Methods: We retrospectively analyzed 20 patients younger than 18 months newly diagnosed with stage 4 MYCN nonamplified neuroblastoma between January 2009 and December 2015. Patients received 9 cycles of chemotherapy and surgery, with or without local radiotherapy, followed by 12 cycles of differentiation therapy with 13-cis-retinoic acid. Chemotherapy consisted of alternating cycles of cisplatin, etoposide, doxorubicin, and cyclophosphamide (CEDC) and ifosfamide, carboplatin, and etoposide (ICE) regimens. Results: The most common primary tumor site was the abdomen (85%), and the most common metastatic sites were the lymph nodes (65%), followed by the bones (60%), liver (55%), skin (45%), and bone marrow (25%). At the end of induction therapy, 14 patients (70%) achieved complete response, with 1 achieving very good partial response, 4 achieving partial response, and 1 showing mixed response. Nine patients (45%) received local radiotherapy. At a median follow-up of 47 months (range, 17-91 months), none of these patients experienced relapse, progression, or secondary malignancy, or died. Three years after chemotherapy completion, none of the patients had experienced grade ${\geq}3$ late adverse effects. Conclusion: Patients younger than 18 months with stage 4 MYCN nonamplified neuroblastoma showed excellent outcomes, without significant late adverse effects, when treated with alternating cycles of CEDC and ICE, followed by surgery and differentiation therapy.

• Journal of Pharmaceutical Investigation
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• v.35 no.1
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• pp.33-38
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• 2005

Recently increasing attention has been focused on solid lipid nanoparticles (SLN) as a parenteral drug carrier due to its numerous advantages that can come from both polymeric particle and fat emulsions, together with the possibility of controlled release and increasing drug stability. Lipophilic drugs such as paclitaxel, cyclosporin A, and all-trans retinoic acid have been successfully entrapped in SLN but the incorporation of hydrophilic drugs in SLN is very limited because of their very low affinity to the lipid. Therefore, as a new approach to improve the loading of hydrophilic drugs, a w/o/w emulsion technique has been developed. The primary objective of the current study was to improve the loading efficiency of a model hydrophilic drug, glycine (Log P = -3.44) into SLN. The proposed preparation process is as follows: A heated aqueous phase consisting of 0.1 ml of glycine solution in water (100 mg/ml), and poloxamer 188 (5 mg) were then added to a molten oil phase containing precirol (100 mg) and lecithin (5 mg). This mixture was dispersed by sonicator, leading to a w/o emulsion. A double emulsion (w/o/w) was formed after the addition of 2% poloxamer solution to the above dispersed system. After cooling the double emulsion, solid lipid nanosuspensions were successfully formed. The lipid nanoparticles had the mean particle size of 441.25 nm, and the average zeta potential of -20.98 mV. The drug loading efficiency was measured to be 8.54% and the drug loading amount was measured to be 0.92%. The w/o/w emulsion method showed an increased loading efficiency compared to conventional o/w emulsion method.

• Environmental Analysis Health and Toxicology
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• v.24 no.2
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• pp.169-178
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• 2009

To investigate the inhibitory effects of Peonia japonica water extract(PJWE) on skin wrinkle formation, skin wrinkles were induced by both the irradiation of UVB and the application of squalene monohydroperoxide to the backs of hairless mice for 4 weeks. And at the same time experimental materials were applied topically. Wrinkles for the control (C) group were formed as a pattern of deep furrows and thick crests. Whereas wrinkles for the positive control (PC, 0.01% retinoic acid) and experimental(E, PJWE) groups were formed as a pattern of shallow furrows and thin crests, which were similar to that of the normal(N) group. Collagen and elastic fibers in dermis of the PC and E groups were almost intact with a regular arrangement, which were similar to those of the N group. The activity of xanthine oxidase, the free radical generating enzyme, was significantly lower in the E group than the C and PC groups. The activities of superoxide dismutase and catalase, the free radical scavenging enzymes, were much higher in the E group than the C and PC groups and similar to the N group. As for the amount of matrix metalloproteinase-3(MMP-3) expression, PC and E groups were significantly lower than the C group. Therefore, PJWE could be very effective natural herbal material for the inhibition or improvement of wrinkle formation in hairless mice skin.

• Environmental Analysis Health and Toxicology
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• v.24 no.2
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• pp.159-167
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• 2009

To investigate the alleviative effects of Peonia japonica water extract(PJWE) on inflammation and skin barrier damage, both the irradiation of UVB and the application of squalene monohydroperoxide (Sq-OOH) to the backs of hairless mice were performed for 4 weeks. And at the same time experimental materials were applied topically. The skin erythema indices for the positive control (PC, 0.01% retinoic acid) and experimental (E, PJWE) groups were lower than that of the control (C) group. Whereas both the lipid and water capacities for the PC and E groups were higher than those of the C group. Epidermis and dermis of the C group were remarkably thickened in comparison with the PC and E groups. Relatively much less number of inflammatory cells, including lymphocytes, neutrophils and macrophages were found in dermis of the PC and E groups compared with the C group. Lipid lamellae of the C group were broken severely showing an irregular arrangement and lipid content was much reduced. Whereas those of the PC and E groups were almost intact with a regular arrangement, which were similar to that of the N group. Taken the results all together, it was confirmed that PJWE could be effective natural herbal material for the alleviation of inflammation and skin barrier damage in hairless mice skin which were induced by UVB and Sq-OOH.

• BMB Reports
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• v.35 no.4
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• pp.377-383
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• 2002

Arsenic trioxide ($As_O_3$) was recently demonstrated to be an effective inducer of apoptosis in patients with relapsed acute promyelocytic leukemia (APL) as well as patients with APL in whom all-trans-retinoic acid and conventional chemotherapy failed. Chronic myelogenous leukemia cells are highly resistant to chemotherapeutic drugs. To determine if $As_O_3$ might be useful for the treatment of chronic myelogenous leukemia, we examined the ability of $As_O_3$ to induce apoptosis in K562 cells. In vitro cytotoxicity of $As_O_3$ was evaluated in K562 cells by a MTT assay: the $IC_50$ value for $As_O_3$ was determined to be $10\;{\mu}m$. When analyzed by agarose gel electorphoresis, the DNA fragments became evident after incubation of the cells with $20\;{\mu}m$ $As_O_3$ for 24 h. We also found morphological changes and chromatin condensation of the cells undergoing apoptosis. Activation of caspase-3 was observed 6 h after treatment with $20\;{\mu}m$ $As_O_3$ by a Western blot analysis. Next, we examined the MAP kinase-signaling pathway of $As_O_3$-induced apoptosis in K562 cells. $As_O_3$ at $10\;{\mu}m$ strongly induced the activation of p38, inhibited $As_O_3$ induced apoptotic cell death. These results suggest that $As_O_3$ is able to induce the apoptotic activity in K562 cells, and its apoptotic mechanism may be associated with the activation of p38.

• Asian Pacific Journal of Cancer Prevention
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• v.17 no.12
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• pp.5139-5145
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• 2016

There has been an increment in the number of studies focused on marine bioactive materials. Many peptides and other biomaterials with anticancer potential have been extracted from various marine animals. Artemia extracts have found uses in sun-light protection cosmetics and anti-aging products. However, contents of biochemical compounds in Artemia spp. and molecular mechanisms of have not been clearly studied in leukemic cells in vitro. In this work, we isolated and purified proteins of Artemia Urmiana. Six clear fractions (A-F) observed on DEAE-cellulose chromatography were assayed for effects on cell growth, differentiation and apoptosis using the human leukemic HL-60 cell line. Cell proliferation analysis by MTT and BrdU assays indicated that did not affect cells, growth. Cells treated with crude extract and fractions A, B and C, but not E and F (up to $100{\mu}g/mL$), exhibited increase of cell growth in a dose dependent manner. Stimulatory effects of fraction D were observed at concentrations of $10{\mu}g/mL$ and above. In nitro blue tetrazolium (NBT) reduction assays, treatment with $100{\mu}g/mL$ of fraction E or F for 96 hr increased the fraction of differentiated cells up to $14.8{\pm}3.56%$ and $16.5{\pm}2.08%$ respectively. Combination of those fractions with retinoic acid had significant synergistic effects on the differentiation of cells ($56.8{\pm}3.7%$ and $67.4{\pm}4.2%$, $p{\leq}0.01$). Annexin-V FITC staining for apoptosis and flow cytometric assays indicated induction of apoptosis by fractions E and F up to 23.8 and 31.8% of cells.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.24
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• pp.10875-10878
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• 2015

The epidermal growth factor receptor (EGFR) is an important surface receptor with N-glycans in its extracellular domain, whose glycosylation is essential for its function, especially in tumor cells. Here, we demonstrated that polylactosamine is markedly increased in H7721 hepatocellular carcinoma cells after treatment with EGF, while it apparently declined after exposure to all-trans retinoic acid (ATRA). In the study of the enzymatic mechanism of this phenomenon, we explored changes in the expression of poly-N-acetyllactosamine (PLN) branching glycosyltransferases using RT-PCR. Among the four glycosyltransferases with altered expression, GnT-V was most elevated by EGF, while GnT-V and B3GNT2 were most declined by ATRA. Next, we conducted co-immunoprecipitation experiments to test whether B3GNT2 and EGFR associate with each other. We observed that EGFR is a B3GNT2-targeting protein in H7721 cells. Taken together, these findings indicated that the altered expression of B3GNT2 will remodel the PLN stucture of EGFR in H7721 cells, which may modify downstream signal transduction.

• Environmental Analysis Health and Toxicology
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• v.23 no.2
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• pp.119-127
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• 2008

The purpose of this study was to investigate the effects of frankincense oil in a skin aging animal model. Skin aging was induced by both the irradiation of UVB and the application of squalene monohydroperoxide (Sq-OOH) to the back of experimental animals for 4 weeks. And at the same time experimental materials were applied topically. Six to seven weeks female SHR-1 hairless mice were divided into five groups including normal (N: saline), control (C: UVB+Sq-OOH+saline), vehicle control (VC: UVB+Sq-OOH+jojoba oil), positive control (PC: UVB+Sq-OOH+0.01% retinoic acid) and experimental (E: UVB+Sq-OOH+3% Frankincense oil) groups, five animals each group. The skin erythema index for the PC and E groups were lower than that of the C group. Whereas, both the lipid and water capacities for the PC and E groups were higher than those of the C group. Wrinkles for the C group were formed as a pattern of deep furrows and thick crests. Whereas, wrinkles for the PC and E groups were formed as a pattern of shallow furrows and thin crests which were similar to that of the N group. As for the both absolute and relative weight of the spleen, the PC group were significantly higher than the other groups. In conclusion, frankincense oil can be used practically for the prevention or improvement of skin aging in terms of health promotion and beauty for the people.

• Environmental Analysis Health and Toxicology
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• v.23 no.2
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• pp.129-138
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• 2008

The purpose of this study was to investigate the effects of frankincense oil in skin aging animal model. Skin aging was induced by both the irradiation of UVB and the application of squalene monohydroperoxide (Sq-OOH) to the back of experimental animals for 4 weeks. And at the same time experimental materials were applied topically. Six to seven weeks female SHR-1 hairless mice were divided into five groups including normal (N: saline), control (C: UVB+Sq-OOH+saline), vehicle control (VC: UVB+Sq-OOH+jojoba oil), positive control (PC: UVB+Sq-OOH+0.01% retinoic acid) and experimental (E: UVB+Sq-OOH+3% Frankincense oil) groups, five animals each group. Lipid lamella and lipid content in stratum corneum of the E group were almost intact with a regular arrangement which were similar to the N group. Collagen fibers in dermis of the E group were almost intact with a regular arrangement which were similar to the N group. Relatively much less number of mast cells and inflammatory cells were found in the E group compared to the C group. The activities of XO, SOD and CAT were no significant difference between the E and N groups. In conclusion, the application of frankincense oil to the skin aging animal model reduced both the generation of free radicals and the damage of skin tissues. Therefore, frankincense oil can be used practically for the prevention or improvement of skin aging in terms of health promotion and beauty for the people.