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        • KCI등재후보

          양수 세포를 이용한 인간배아줄기세포의 배양

          김희선,설혜원,안희진,오선경,구승엽,김석현,최영민,김정구,문신용,Kim, Hee-Sun,Seol, Hye-Won,Ahn, Hee-Jin,Oh, Sun-Kyung,Ku, Seung-Yup,Kim, Seok-Hyun,Choi, Young-Min,Kim, Jung-Gu,Moon, Shin-Yong 대한생식의학회 2004 Clinical and Experimental Reproductive Medicine Vol.31 No.4

          Objective: This study was performed to evaluate the possibility of prolonged culture of human embryonic stem cells (hESC; SNUhES2) on human amniotic fluid cells (hAFC), which had been storaged after karyotyping. Method: The hAFC was prepared for feeder layer in the presence of Chang's medium and STO medium (90% DMEM, 10% FBS) at $37^{circ}C$ in a 5% $CO_2$ in air atmosphere. Prior to use as a feeder layer, hAFC was mitotically inactivated by mitomycin C. The hESCs on hAFC were passaged mechanically every seven days with ES culture medium (80% DMEM/F12, 20% SR, bFGF). Results: The hAFC feeder layer support the growth of undifferentiated state of SNUhES2 for at least 59 passages thus far. SNUhES2 colonies on hAFC feeder appeared slightly angular and flatter shape as compared with circular and thicker colonies observed with STO feeder layer and showed higher level with complete undifferentiation in seven days. Like hESC cultured on STO feeders, SNUhES2 grown on hAFC expressed normal karyotype, positive for alkaline phosphatase activity, high telomerase activity, Oct-4, SSEA-3, SSEA-4, Tra-1-60 and Tra-1-81 and formed embryoid bodies (EBs). Conclusion: The hAFC supports undifferentiated growth of hESC. Therefore, these results may help to provide a clinically practicable method for expansion of hESC for cell therapies.

        • KCI등재후보
        • KCI등재후보

          환경 방사선 생물학적 감시 지표로서 야생 등줄쥐의 방사선 감수성

          김희선,니시무라 요시카즈,김종순,Kim, Hee-Sun,Nishimura, Y.,Kim, Chong-Soon 한국방사성폐기물학회 2006 방사성폐기물학회지 Vol.4 No.1

          이 연구에서는 원자력발전소 주변 환경 방사선이 생태에 미치는 영향을 감시할 수 있는 생물학적 지표로서 야생 등줄쥐의 활용 가능성을 평가하였다. 국내 5지 역을 선정하여 10월에 한정하여 등줄쥐를 채집하였는데, 농경지에서 산중턱에 이르기까지 폭 넓게 서식하였다. 채집한 쥐들에 대하여 황갈색 피모와 검은색 등줄무늬를 관찰하였는데, 외부형태 특성 가운데 몸통의 길이, 꼬리의 길, 귀의 길이를 계측한 결과, 등줄쥐의 분류기준과 일치하였다. 아울러, 간장내 효소형을 분석한 결과 국내에 서식하는 대부분의 등줄쥐가 Apodemus agrarius라는 것을 알 수 있었다. 또한, 야외에서 생포한 등줄쥐를 암수 한 마리씩 동거시키고 생산된 새끼가 5주령에 도달하였을때 방사선을 조사하고, ICR 마우스를 비교로 생존율과 적혈구내 미소핵 출현빈도를 분석한 결과, 반치사 선량$(LD_{50/30})$은 5와 7.9Gy였다. 이 연구결과로 야생 등줄쥐가 원자력 발전소 주변 수준의 방사선이 인간생환에 미치는 영향을 판단할 수 있는 생물학적 인 지표로서 잠재적 활용성이 높다는 것을 알 수 있었다. This study examined the possibility of using dark-striped field mice as a biological indicator for the environmental radio-surveillance. For this study, dark-striped field mice were caught from five areas of Kyonggi, Kyongsang, Chungchong and Cholla provinces. The external morphological characteristics and isoenzymic types of dark-striped field mice were studied after they were captured. Among the external morphological characteristics, the dark-brown coat, dark back stripe, head-to-tail length, tail length, and ear length matched the taxonomical characteristics of dark-striped field mice. The analyses on L-lactate dehydrogenase, aspartate aminotransferase, and malate dehydrogenese revealed that one species of dark-striped field mice, called Apodemus agrarius, was inhabitated throughout a wide range of Korea. On the other hand, the frequency of micronuclei in peripheral polychromatic erythrocytes to survived mice after irradiation also analyzed. The LD50/30 of A. cgrarius and ICR mice were approximately 5 Gy and 7.9Gy, respectively. The results of the study reveal that wild A. asrarius have a high potential as a biological monitoring system to determine the impact of radiation in areas such as those within the vicinity of nuclear power plants.

        • SCOPUSKCI등재

          Molecular Cloning and Expression of the Acetyl Xylan Esterase Gene(estII) of Bacillus Stearothermophilus in Escherichia coli

          김희선,엄수정,조쌍구,최용진,Kim, Hee-Sun,Eom, Soo-Jung,Cho, Ssang-Goo,Choi, Yong-Jin 한국미생물 · 생명공학회 1994 한국미생물·생명공학회지 Vol.22 No.6

          Bacillus stearothermomophilus, a strong xylan degrader, was confirmed to express multiple esterase activities in addition to the major xylanolytic enzymes. One of the genes encoding the esterases was isolated from the genomic library of B. stearothermophilus constructed with EcoRl restriction endonuclease and pBR322 plasmid. Three recombinant plasmids showing the tributyrin degrading activity were selected from approximately 7, 000 E. coli HB101 transformants, and were found to have the same insert of a 3.2 kb DNA fragment. Restriction mapping and hybridization studies revealed that the gene(estII) on the hybrid plasmid (pKMG7) had originated from the B. stearothermophilus chromosome, and was distinct from the estl, another esterase gene of B. stearothermophilus isolated in the previous work. The E. coli cells harboring pKMG7 produced an acetylxylan esterase that exibited similar substrate specificity to the esterase encoded by the estI gene.

        • SCOPUSKCI등재후보

          Subfertile 정자처리에 Modified Mini-Percoll 방법의 효용성에 관한 연구

          김희선,장현정,이희옥,이숙현,이상훈,Kim, Hee-Sun,Chang, Hyun-Jung,Lee, Hee-Ok,Lee, Sook-Hyun,Lee, Sang-Hoon 대한생식의학회 1994 Clinical and Experimental Reproductive Medicine Vol.21 No.3

          Semen preparation for Assisted Reproductive Technology(ART) involves the separation of the motile sperm from non-motile, dead sperm, cellular debris and seminal fluid. The aim of this study is to compare the motile sperm recovery rate and motility index of different sperm preparation method(swim-up method, 80% isotonic continuous percoll method, two-layer discontinuous percoll method, mini-percoll method). In Mini-Percoll method, pellets from patients were suspended in 0.3ml of medium and layerd on a discontinuous percoll gradient consisting of 0.3ml each of 50, 70, 95% isotonic percoll. All semen samples are divided into normal and subnormal sperm group(oligo-, astheno-, oligoasthenozoospermia). Especially, we evaluated the effect of mini-percoll method in subnormal sperm group. In normal sperm group, mini-percoll method and two layer discontinuous percoll method (40%/80%) allowed increasing of motile sperm recovery rate. But motility index was higher in swimup method than the other methods. In subnormal sperm group, mini-percoll method has advantages as compare with the other methods in motile sperm recovery rate and motility index. These results suggest that modified mini-percoll method could be certainly a valuable tool in some cases of severe male factor sperm.

        • 근골격계 종양의 종류에 따른 케모카인 유전자의 발현

          김희선,백원진,이원재,신덕섭,Kim, Hee-Sun,Baik, Won-Jin,Lee, Won-Jae,Shin, Duk-Seop 대한근골격종양학회 2003 대한골관절종양학회지 Vol.9 No.2

          목적: 근골격계 종양의 종류에 따른 케모카인 발현 양상을 보고, 원발성 골육종과 재발된 골육종, 골육종의 항암화학요법 전과 후의 발현 차이가 있는지를 조사하고자 한다. 대상 및 방법: 종양조직을 얻을 수 있었던 원발성 양성 및 악성 근골격계 종양 10예, 재발된 골육종 1예, 화학요법 후의 골육종 1예 및 정상조직의 대조군 1예를 대상으로 하였다. RNA를 분리한 다음, 역전사 중합효소 연쇄반응(RT-PCR )과 RNase protection assay (RPA)를 이용하여 싸이토카인과 케모카인의 발현을 조사하였다. 각각의 종양간에 발현도 차이가 있는지를 알기 위하여 SPSS를 사용한 Fisher's exact test를 이용하여 통계적 처리를 하였다. 결과: IL-8과 TNF-${\alpha}$는 모든 조직에서 발현되었고, IFN-${\gamma}$는 2예 외에 모두 발현하였으며, RANTES의 경우 연부조직 종양 5예, 골종양 4예에서, GRO${\alpha}$는 연부조직 종양 1예, 골종양 2예에서 발현되었고, MCP-1과 IP-10의 경우 악성 골종양군에서는 2예만, 나머지에서는 모두 발현되는 다양한 양상을 보였다. 원발성 골육종에 비해 재발된 골육종 조직에서의 발현은 IFN-${\gamma}$를 제외하고, 전반적으로 발현도가 훨씬 강하였으며, 모든 종류의 케모카인 및 사이토카인들이 발현되었다. 화학요법 후에는 RANTES와 IFN-${\beta}$만이 발현되었으며, TGF${\beta}$ isoform 중 TGF${\beta}_1$ 유전자의 발현만 관찰되었다. 결론: 대상의 수가 적어 통계적 분석을 통한 정확한 자료의 제시는 불가능하였으나, 연부조직 종양과 골종양에 있어 일부 케모카인 및 사이토카인의 발현 차이가 있음을 확인하였고, 골육종의 경우 IFN-${\gamma}$ 및 TGF-${\beta}$ isoform의 발현 분석이 종양의 재발과 치료 후 변화에 대한 지표에 응용될 가능성이 있음을 제시하였다. Purpose: The current study was designed to investigate the expression pattern of chemokine in musculoskeletal tumors, and between primary osteosarcoma and recurred, and postchemotherapy one. Materials and methods: Ten primary soft tissue and bone tumors, one primary, one recurred, one post-chemotherapy osteosarcoma, and one normal control patients were included in the current study. RT-PCR and RPA were used for the investigation of the expression of cytokines and chemokines. Fisher's exact test in SPSS was used for the statistical analysis. Results: IL-8 and TNF-${\alpha}$ were expressed in all tumor tissues, IFN-${\gamma}$ was in all except two cases, RANTES was in 5 soft tissue tumors and 4 bone tumors, GRO-${\alpha}$was in one soft tissue tumor and 2 bone tumors, and MCP-1 and IP-10 were in two bone tumors and in all the other group. In recurred osteosarcoma all the cytokines and chemokines were expressed, and the degree of the expression was stronger than the primary, except IFN-${\gamma}$. After chemotherapy, RANTES, IFN-${\beta}$ and TGF${\beta}_1$ among the TGF${\beta}$isoforms were expressed. Conclusion: There were differences in the expression of cytokines and chemokines in some different bone and soft tissue tumors, even though it was impossible to support this statistically due to small numbers of cases. The expression pattern of IFN-${\gamma}$and TGF-${\beta}$ isoform in osteosarcoma could be used for the study of tumor recurrence and the changes after chemotherapy.

        • SCOPUSKCI등재

          국내 개의 리켓치아 감염에 관한 역학조사

          김희선,강문일,이채용,김성호,Kim, Hee-sun,Kang, Mun-il,Lee, Chai-yong,Kim, Sung-ho 대한수의학회 1995 大韓獸醫學會誌 Vol.35 No.4

          One hundred twelve domestic dog sera were collected from Kyonggi, Kanngwon, Cholla, and Kyongsang province. All the sera were examined by immunofluorescence antibody test(IFAT) against scrub typhus, murine typhus and spotted fever group rickettsia infection. The antibody prevalence was 5.40%(6/112) against scrub typhus. The sero-prevalence rates of scrub typhus were 16.30%(5/31) in Cholla and 5.90%(1/17)in Kyonggi areas. The prevalence of scrub typhus was higher in older dogs that were 6 years or more. The antibody prevalence was 8.04%(9/112) against murine typhus. The sero-positive rates to murine typhus were 5.88%(1/17) in Kyonggi, 8.33%(2/24) in Kangwon, 9.68%(3/31) in Cholla, and 7.50%(3/40) in Kyongsang areas, respectively. The sero-positive dogs to murine typhus were widely distributed throughout all age groups from 2 months to above 9 years old. The antibody prevalence was 16.96%(19/112) for spotted fever group rickettsial infection. The sero-positive rates of spotted fever group rickettsia were 11.76%(2/17) in Kyonggi, 12.90%(4/31) in Cholla, and 32.50%(13/40) in Kyongsang areas. The sero-positive dogs to spotted fever group rickettsia were observed at all age groups of dog. The sero-positive prevalence between sex showed higher in female than in male except for spotted fever group rickettsia infection.

        • SCOPUSKCI등재후보

          생쥐 8세포배의 Compaction에 미치는 칼슘 Inhihitor의 영향

          김희선,배인하,Kim, Hee-Sun,Bae, In-Ha 대한생식의학회 1994 Clinical and Experimental Reproductive Medicine Vol.21 No.1

          In mammalian development, the embryo which is in the process of compaction, involves a progressive flattening of blastomeres against each other with the consequence that the embryo assumes a spherical shape. This stage happens in the first differentiation. The present study was aimed to examine the involvement of other metal ions in compaction by treating with various divalent cations in the absence of $Ca^{++}$. When 8-cell embryos were cultured in $Ca^{++}$-free medium for 24hrs, they developed to 16-cell stage but did not compaction, and degenerated after 48hrs of culture. Embryos were cultured in $Ca^{++}$-free medium for 24hrs and then transferred to the control medium showed the normal compaction afterwards. When 8-cell embryos were cultured in the presence of $Ni^{++}$, known as a $Ca^{++}$ inhibitor, they cleaved to 16-cell stage but did not compact in the absence of $Ca^{++}$. On the other hand, embryos cultured in the media containing both $Ca^{++}$ and $Ni^{++}$ developed normally so that they underwent compaction during culture for 48hrs. However, they failed to hatch during further 24hrs in the same medium, indicationg that $Ni^{++}$ may exert some harmful effects. Embryos grow in the control medium that contained $Ca^{++}$ but not $Ni^{++}$, developed to the hatched blastocysts. The treatment with $Cd^{++}$ $10^{-1}$,$10^{-2}{\mu}M$, $Mn^{++}$ or $Ba^{++}$ 10,100, $1000{\mu}M$ in $Ca^{++}$-free medium, respectively, inhibited compaction and embryonic degeneration began as in $Ca^{++}$-free medium. When 3, 5, 10mM of $Sr^{++}$, known as a substitute for $Ca^{++}$ in cell, was added to $Ca^{++}$-free medium, respectively, compaction was induced unlike the above metal ions. Embryos were cultured in $Sr^{++}$ developed to blastocysts, but failed to hatch after 72hrs and degenrated. On the other hand, when embryos were cultured in 3, 5, 10mM of $Sr^{++}$ but in $Ca^{++}$-free medium for 24hrs respectively and then transferred to the control, they showed the similiar development as that in the control.

        • KCI우수등재

          전단경간비 및 축력에 따른 깊은 보의 거동 특성에 대한 연구

          김희선,박민혜,이민섭,신영수,Kim, Hee-Sun,Park, Min-Hye,Lee, Min-Seob,Shin, Yeong-Soo 대한건축학회 2011 대한건축학회논문집 Vol.27 No.6

          The purpose of this study is to investigate effect of axial loads on deep RC beams. Toward this goal, deep RC beams having different shear span-to-depth ratios are fabricated and bending tests are performed on the beams applied with different magnitudes of axial loads. From the tests, strains on various locations, beam deflections, beam compression and shear strains are obtained. The experimental results show that, for the most of the tested beams, failure due to shear is occurred in the axially loaded deep RC beam and the axial load works beneficially to structural performance. However, when the shear span-to-depth ratio is sufficiently small and the large axial load is applied, the beam is failed earlier than as expected due to bearing failure.

        • KCI등재
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