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      • KCI등재

        제1형 당뇨병 쥐 모델에서 유전공학적 제조 K-세포 이식을 통한 당뇨병의 치료

        심주연 ( Ju Yeon Sim ),김주희 ( Ju Hee Kim ),안유배 ( Yu Bae Ahn ),송기호 ( Ki Ho Song ),한제호 ( Je Ho Han ),차봉연 ( Bong Yun Cha ),이숙경 ( Sook Kyung Lee ),문성대 ( Sung Dae Moon ) 대한당뇨병학회 2009 Diabetes and Metabolism Journal Vol.33 No.6

        연구배경: 인슐린 유전자 치료법의 이상적인 표적세포로 K-세포가 알려져 있다. 이전 연구에서 본 연구자들은 EBV-유래 에피솜 벡터를 이용하여 K-세포에서 포도당농도 의존적인 인슐린 분비가 이루어지는 것을 실험실 환경에서 확인한 바 있다. 본 연구에서는 스트렙토조토신(STZ)으로 유발된 제1형 당뇨병 쥐의 신장 피막에 유전공학적으로 제조된 K-세포를 이식하여 당뇨병이 치료되는지를 관찰해 보았다. 방법: 인슐린이 분비되도록 제조된 K-세포를 STZ으로 유도된 BALB/c Nude 마우스의 신장 피막에 이식한 후 혈당과 몸무게를 측정하였다. 마우스는 정상군과 STZ으로 유도된 당뇨병군 그리고 당뇨병군의 일부는 K-세포를 이식한 치료군 등으로 분류하여 실험을 하였다. 이식 4주 후 모든 마우스에서 6시간 동안 금식한 후 2 g/kg의 포도당을 복강내로 주사하여 당내성검사를 하였다. 당내성검사가 끝나자마자 모든 마우스의 신장과 췌장을 적출하여 인슐린, 글루카곤, C-peptide 등으로 면역조직화학염색 및 면역형광염색을 한 후 그 결과를 분석하였다. 결과: STZ을 복강에 주사한 마우스는 주사 3일 후 혈당이 300±50 mg/dL로 상승하였다. 정상군과 K-세포를 이식한 당뇨병군에서는 몸무게가 점차 증가하였으나, STZ만을 주사한 당뇨병군에서는 실험기간 동안 낮게 유지되었다. 그리고 정상군에서는 정상범위의 혈당분포를 유지하였으나, STZ만을 주사한 당뇨병군에서는 지속적으로 혈당이 높게 유지되었다. 그리고 K-세포를 이식한 당뇨병군에서는 STZ만을 주사한 군에서보다 혈당이 서서히 떨어졌으며 이식 2주 후부터는 급격히 감소하였다. 당부하검사 결과 정상군과 K-세포를 이식한 당뇨병군에서는 서로 유사한 내당력을 보였으나 STZ만을 주사한 군에서는 지속적으로 포도당 불내성을 보였다. 조직면역검사 결과 정상군의 췌장에서는 인슐린과 글루카곤, C-peptide 모두에 염색이 되었지만 STZ만을 주사한 마우스의 췌장에서는 췌도파괴로 인슐린은 물론 글루카곤, C-peptide 모두에 염색이 되지 않았다. 그러나, K-세포를 이식한 당뇨병군의 신장에서는 면역조직화학염색에서 인슐린과 C-peptide 모두에서 염색이 되었으며 면역형 광염색에서도 인슐린이 발현하는 것을 관찰할 수 있었다. 결론: 유전공학적으로 제조된 K-세포가 마우스 신장 피막에 착상되어 혈당농도 의존적으로 인슐린을 분비하여 당뇨병이 치료되는 것을 생체 내에서도 확인할 수 있었으며, EBV-유래 에피솜벡터를 이용한 유전공학적 조제 K-세포는 제1형 당뇨병의 치료를 위한 대체베타세포로 사용될 수 있을 것으로 생각된다. Background: K-cells function as targets for insulin gene therapy. In a previous study, we constructed EBV-based plasmids expressing rat preproinsulin controlled by glucose-dependent insulinotropic polypeptide promoters. In the present study, we attempted to correct hyperglycemia in vivo using genetically engineered K-cells in a mouse model of type 1 diabetes. Methods: K-cells expressing insulin were transplanted under the kidney capsules of STZ-induced diabetic mice. The blood glucose levels and body weights of the experimental animals were measured daily. After four weeks, the mice were injected intra-peritoneally with 2 g/kg glucose following a 6 hr fast. Blood glucose levels were measured immediately following glucose injections. All animals were sacrificed at the end of the glucose tolerance study, and pancreas and graft-bearing kidney tissue samples were stained with antibodies against insulin, glucagon, and C-peptide. Results: The body weights of K-cell-transplanted diabetic mice increased after transplantation, whereas those of untreated diabetic control mice continued to decline. The blood glucose levels of K-cell-transplanted diabetic mice decreased gradually during the two weeks following transplantation. After intra-peritoneal injection of glucose into K-cell-transplanted diabetic mice, blood glucose levels increased at 30 minutes, and were restored to the normal range between 60 and 90 minutes, while untreated control diabetic mice continued to experience hyperglycemia. Kidney capsules containing transplanted K-cells were removed, and sections were stained with anti-insulin antibodies. We detected insulin-positive cells in the kidney capsules of K-cell-transplanted diabetic mice, but not in untreated control mice. Conclusion: We detected glucose-dependent insulin secretion in genetically engineered K-cells in a mouse model of type 1 diabetes. Our results suggest that genetically modified insulin producing K-cells may act as surrogate β-cells to effectively treat type 1 diabetes. (Korean Diabetes J 33:466-474, 2009)

      • 갯기름나물 중 Ethoprophos와 Metaldehyde의 잔류 특성

        심주연 ( Ju Yeon Sim ),조성용 ( Sung Yong Jo ),김용범 ( Yong Beom Kim ),황준혁 ( Jun Hyuk Hwang ),강은옥 ( Eun Ok Kang ),경기성 ( Kee Sung Kyung ) 한국환경농학회 2020 한국환경농학회 학술대회집 Vol.2020 No.-

        This study was carried out to investigate the residual characteristics of insecticide ethoprophos and metaldehyde in coastal hogfennel under greenhouse conditions. The test commercial pesticides, ethoprophos 5% GR and metaldehyde 6% GR were treated onto upland soil surface at the recommended and double dose before planting and then the test crop was transplanted. Sampling was done on the pre-harvest day of 50 days after treatment. To investigate residue change after harvest, additional samples were also collected 7 and 14 days after harvest. Limits of quantitation of ethoprophos and metaldehyde in the crop were all 0.01 mg/kg. Recoveries of ethoprophos and metaldehyde were ranged from 101.8 to 110.2% and from 71.1 to 93.0%, respectively. The residues of ethoprophos in the crop collected at harvest in recommended dose and double dose plots were 0.42 and 0.61 mg/kg, respectively. In case of metaldehyde, those were 0.25 and 1.37 mg/kg, respectively. Also, residues in the test crop collected 7 and 14 days after harvest found to be decreased comparing to those in the crop at harvest. The dissipation rates of the test pesticides in the crop collected 7 and 14 days after harvest were from 60.7 to 61.9% and from 51.1 to 56.0%, respectively, in both recommended and double dose.

      • KCI등재
      • 갯기름나물 중 Hexythiazox와 Fenpropathrin의 잔류 특성

        김용범 ( Yong Beom Kim ),조성용 ( Sung Yong Jo ),심주연 ( Ju Yeon Sim ),황준혁 ( Jun Hyuk Hwang ),강은옥 ( Eun Ok Kang ),경기성 ( Kee Sung Kyung ) 한국환경농학회 2020 한국환경농학회 학술대회집 Vol.2020 No.-

        This study was carried out to investigate dissipation patterns of hexythiazox and fenpropathrin in coastal hogfennel. The test commercial hexythiazox 10% WP and fenpropathrin 5% EC were diluted 2,000- and 1,000-time, respectively, and then the diluent were sprayed 2 times with 7-day intervals before harvest onto the test crop. Samples were collected at 0, 3, 5, 7 and 14 days after the last spraying. Limits of quantitation (LOQs) of the test pesticides were all 0.01 mg/kg. Recoveries of hexythiazox and fenpropathrin at 10- and 50-fold levels of their LOQs ranged from 80.3 to 96.3% and from 90.6 to 93.8%, respectively. The highest residues of hexythiazox and fenpropathrin in the crops collected just after spraying were 2.30 and 2.02 mg/kg, respectively, whereas those in the samples collected 14 days after harvest were 0.12 and 0.15 mg/kg, respectively. Also, half-lives of hexythiazox and fenpropathrin in coastal hogfennel were 3.1 and 3.7 days, respectively, indicating that the amount of hexythiazox and fenpropathrin in coastal hogfennel were time-coursely decreased.

      • 건조과정을 통한 구기자의 Benomyl 잔류량 변화의 확인에 관한 연구

        조성용 ( Sung Yong Jo ),김용범 ( Yong Beom Kim ),심주연 ( Ju Yeon Sim ),황준혁 ( Jun Hyuk Hwang ),강은옥 ( Eun Ok Kang ),경기성 ( Kee Sung Kyung ) 한국환경농학회 2020 한국환경농학회 학술대회집 Vol.2020 No.-

        This study was carried out to determine effect of drying on benomyl residue in Chinese matrimony vine. The test commercial pesticide, benomyl 50% WP, was diluted 2,000-fold and then the diluted solution was sprayed 2 times onto Chinese matrimony vine at 7-day intervals. Samples were collected after last spraying (0 day), 7 and 14 days and then dried with a hot-air oven to make the dried product. Residue of benomyl was calculated by conversion of carbendazim obtained to benomyl on the bais of their molecular weight. The limits of quantitation of carbendazim were all 0.01 mg/kg. The recoveries of carbendazim in fresh and dried product ranged from 87.4 to 104.2% and from 72.6 to 75.4%, respectively. The amounts of benomyl residues in fresh was slowly decreased after spraying. The residual amounts of benomyl in fresh and dried product at day 0 were 0.89 and 1.14 mg/kg, indicating that benomyl residue was increased in the dried product by removing of water.

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