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Photodegradable hydrogels for single cell research
신동식 한국공업화학회 2015 한국공업화학회 연구논문 초록집 Vol.2015 No.0
A method for capture, analysis, and sorting of live cells using a photodegradable hydrogel (photogel) functionalized with antibodies will be demonstrated. Photogel substrates used to first isolate human CD4 or CD8 T-cells from a heterogeneous cell suspension and then to release desired cells or groups of cells by UV-induced photodegradation. Flow cytometry analysis of the retrieved cells revealed approximately 95% purity of CD4 and CD8 T-cells, suggesting that this substrate had excellent specificity. To demonstrate the possibility of sorting cells according to their function, photogel substrates that were functionalized with anti-CD4 and anti-TNF-a antibodies were prepared. Single cells captured and stimulated on such substrates were identified after immunofluorescent staining and could be retrieved by site-specific exposure to UV light through a microscope objective. The use of a photodegradable gel allows the detection of cells of interest based on their function and the release of these cells for downstream analysis, which could entail gene expression studies or continued cultivation.
신동식,Sin, Dong-Sik 대한석유협회 1995 석유와 에너지 Vol.1995 No.6
이 자료는 지난 5월 3일 한국종합전시장에서 열린 한국자동차공업협회 주최의「자동차환경정책」세미나에서「국제환경규제동향과 자동차산업정책방향」제목으로 발표된 내용 중 발췌한 것이다. <편집자 주>
신세포암 동물모델에서 전신적 면역치료와 동반한 국소 냉동요법의 항암 효과
신동식,고영휘,최훈,강석호,배재현,박홍석,문두건,천준,윤덕기 대한비뇨의학회 2008 Investigative and Clinical Urology Vol.49 No.11
Purpose: To investigate synergistic effect of local cryoablation with systemic immunotherapy, the tumor control ability and immunologic responses of combining these two modalities was compared with that of cryoablation, surgical excision, and immunotherapy only group in a tumor re-challenge model. Materials and Methods: Preliminary experiments were performed in two stages. The first stage consisted of 36 Balb/c mice with Renca bearing tumors imbedded in the right thigh, and was treated with interleukin-2 (IL-2) and interferon-alpha(IFN-α) to evaluate the efficacy of immunotherapy and to determine the adequate dosage. The second stage was performed on 10 mice, to evaluate histological changes and efficacy after cryoablation. The main experiment was performed on 48 mice, divided into 6 groups of control with tumor implantation, excision of tumor, excision combined with immunotherapy, cryoablation of tumor, cryoablation with immunotherapy and control without tumor. After treatment, tumor re-challenge was performed with Renca cell, then the growth pattern was evaluated with physical measurements, and immune response was investigated with fluorescent activated cell sorter and cytotoxicity assay. Results: Preliminary studies on immunologic efficacy revealed that IL-2 and IFN-α have a dose dependent inhibition of tumor growth. The main experiment evaluating the efficacy of combination treatment revealed that cryoablation with immunotherapy proved to be most effective in terms of tumor recurrence and tumor growth inhibition, yet the difference was not statistically significant from monotherapy with cryoablation. However, cytotoxicity was significantly increased cryoablation with immunotherapy compared with other groups. Conclusions: Cryoablation on tumor re-challenge mice model showed advantages with immunotherapy most prominently in cytotoxicity. Purpose: To investigate synergistic effect of local cryoablation with systemic immunotherapy, the tumor control ability and immunologic responses of combining these two modalities was compared with that of cryoablation, surgical excision, and immunotherapy only group in a tumor re-challenge model. Materials and Methods: Preliminary experiments were performed in two stages. The first stage consisted of 36 Balb/c mice with Renca bearing tumors imbedded in the right thigh, and was treated with interleukin-2 (IL-2) and interferon-alpha(IFN-α) to evaluate the efficacy of immunotherapy and to determine the adequate dosage. The second stage was performed on 10 mice, to evaluate histological changes and efficacy after cryoablation. The main experiment was performed on 48 mice, divided into 6 groups of control with tumor implantation, excision of tumor, excision combined with immunotherapy, cryoablation of tumor, cryoablation with immunotherapy and control without tumor. After treatment, tumor re-challenge was performed with Renca cell, then the growth pattern was evaluated with physical measurements, and immune response was investigated with fluorescent activated cell sorter and cytotoxicity assay. Results: Preliminary studies on immunologic efficacy revealed that IL-2 and IFN-α have a dose dependent inhibition of tumor growth. The main experiment evaluating the efficacy of combination treatment revealed that cryoablation with immunotherapy proved to be most effective in terms of tumor recurrence and tumor growth inhibition, yet the difference was not statistically significant from monotherapy with cryoablation. However, cytotoxicity was significantly increased cryoablation with immunotherapy compared with other groups. Conclusions: Cryoablation on tumor re-challenge mice model showed advantages with immunotherapy most prominently in cytotoxicity.
Development of the Method Detecting Cell-Secreted Proteases
신동식 한국공업화학회 2018 한국공업화학회 연구논문 초록집 Vol.2018 No.0
A method for monitoring protease activity using redox-labeled peptides functionalized onto gold electrodes will be described. Existing bioanalytical approaches for detection of protease expression from cancer cells include immunoassay and gelatinase zymography that are based on the end point analysis. Our biosensor consisted of a micropatterned surfaces with miniature electrodes positioned next to antibodycontaining cell capture sites. Protease molecules secreted by captured cells cleave peptide segment with redox label, causing the electrochemical signal to decrease. This principle was used to construct methylene blue (MB)-modified peptides specific to MMP9, then capture monocytes (U937 cell line) and monitor protease release/activity from the cells. The method will be providing numerous real time secretion profiles rather than conventional detection methods including ELISA, PCR and gel electrophoresis.