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TPA로 야기된 HL-60 세포의 기질부착에 대한 Asadisulphide의 억제효과
유관희(Kwan-Hee You),박미아(Mi-A Park),김선희(Seon-Hee Kim),안병준(Byung-Zun Ahn) 대한의생명과학회 2000 Biomedical Science Letters Vol.6 No.3
항암제로 사용되어 온 아위를 사용하여 극성에 따른 용매추출 분획을 얻고 TPA로 처리 된 HL-60 세포를 사용하여 기질부착 억제실험을 수행한 결과, ethyl acetate (EA) 층에서 가장 강한 부착억제효과가 있음을 알아 내었기에 ethyl acetate (EA)층을 다시 ethyl acetate (EA), hexane (HEX), ethyl ether (EE)로 추출한 뒤 3회 chromatography하여 아위 순수활성물질인 asadisulphide를 분리해 내었다. 이 순수활성물질을 가지고 HL-60 암세포에 대한 부착 억제실험을 수행한 결과 asadisulphide는 최소농도 2 ㎍/㎖에서 HL-60 세포의 부착을 98%억제하였다. 또한 assdisulphide는 HL-60 세포에 대해 ED??값은 2.6㎍/㎖이었으며, 20㎍/㎖ 농도 이하에서 세포독성이 없고 항암효과가 있다는 사실을 규명하였다. Asadisulphide were purified from Ferrula assafoetida by organic soivent extraction and chromatography. Its inhibitory effects on the TPA-induced adherence of HL-60 cells was analyzed. Since ethyl acetate extracts of F. assafoetida has the strongest inhibitory effect on adherence of HL-60 cells, it was re-extracted with ethyl acetate, hexane, and ethyl ether and chromatographed three times to isolate asadisulphide. At the minimum concentration of 2 ㎍/㎖, asadisulphide inhibited adherence of 98% of HL-60 cells that have been treated with TPA. It also showed anti-cancer effect with no cytotoxity in the ED?? value of 20 ㎍/㎖.
랫드 수초좌골신경섬유에서 Neurofascin분포에 대한 면역세포화학적 연구
장병화,유관희,이종환,조익현,배춘식,박창현,한정미,최농훈,장병준,Chang, Byung-Hwa,You, Kwan-Hee,Lee, Jong-Hwan,Cho, Ik-Hyun,Bae, Chun-Sik,Park, Chang-Hyun,Han, Jeong-Mi,Choe, Nong-Hoon,Chang, Byung-Joon 한국현미경학회 2006 Applied microscopy Vol.36 No.2
Neurofascin은 L1CAM의 하나로 신경섬유의 발달과정에서 중요한 역할을 하는 것으로 알려져 있다. 말초신경의 수초형성과 관련된 neurofascin의 역할을 알아볼 목적으로 면역형광염색과 면역전자현미경기법을 이용하여 랫드의 수초좌골신경섬유에서 neurofascin의 분포를 추구하여 다음과 같은 결과를 얻었다. 1.수초형성이 진행됨에 따라 좌골신경섬유에서 neurofascin 분포는 매우 심하게 변화되었다. 2. 수초신경섬유에서 neurofascin은 Ranvier마디에서 약하게 국재하였다. 3. Neurofascin은 수초신경섬유의 paranodal loop, Schmidt-Lantermann incisure, 속축삭사이막, 바깥축삭사이막처럼 Schwann세포의 막이 밀착되지 않은 부위에서도 뚜렷하게 국재하였다. 이상의 연구결과로 neurofascin은 Schwann세포의 마주보는 막사이에 이상적인 간격을 유지하는데 어떤 역할을 하는 것으로 생각되며, 수초층에서 물질이동이 가능하게 하는 것으로 보인다. Neurofascin, one of the members of L1CAM, has been known to have some important roles during the development of nerve fibers. In order to investigate the role of neurofascin associated with the myelination of peripheral nerves, the localization of neurofascin in myelinated rat sciatic nerve fibers was studied with the immuno-fluorescence and immune-electron microscopy and the results are as follows; 1. According to the myelination is going on, neurofascin localization was dramatically changed in the sciatic nerve fibers. 2. In the myelinated fibers, neurofascin was weakly localized along the axolemma at the node of Ranvier. 3. Neurofascin was also apparantly localized at the non-compact area of Schwann cell membrane such as paranodal loop, Schmidt-Lantermann incisure, inner & outer mesaxons in the myelinated fibers. From the above results, neurofascin is likely to have a role to sustain the ideal gap of apposing membranes of Schwann cell, so it may enable to materials transport in the myelin sheath.
쓰름매미 (Meimuna mongolica D.) 복안의 전자현미경적 관찰
최춘근,유관희,신길상,최임순,Choi, Choon-Keun,You, Kwan-Hee,Shin, Kil-Sang,Choe, Rim-Soon 한국현미경학회 1979 Applied microscopy Vol.9 No.1
Electron microscopic studies were carried out to investigate the fine structure of ommatidia of the compound eyes in Meimuna mongolica D. The crystalline cone appears round, and is composed of four double-membraned cone cells and surrounded by pigment cells having many pigment granules. The rhabdom is a closed type, and is composed of four rhabdomeres Its cross section reveals lamellated microvilli which are oriented in four different directions suggesting that these represent four photoreceptive sites. The microvilli, in a cross sectional view, are hexagonal in shape with a central axis inside. There are usually eight retinular cells arranged radially from the rhabdom, but in some retinular layers seven or nine retinular cells could be observed. The cytoplasm of each retinular cell is interconnected with that of microvilli of the rhabdomere, but the appearance of this interconnection varies depending on the number of retinular cells.'The retinlilar cells neighboring the microvilli seem to have well-developed perirhabdomal vacuoles and mitochondria as well as pigment granules surrounding these vacuoles.
백혈병 세포주에 대한 (+/-)-ar-Turmerone, 자근 및 황금추출물에 의한 항암제의 세포독성 증강효과
이윤영(Yun Young Lee),유관희(Kwan Hee You),김삼용(Sam Yong Kim),안병준(Byung Zun Ahn) 대한약학회 1991 약학회지 Vol.35 No.3
Using the calorimetric [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] (MTT)assay, we evaluated the chemosensitivity of 8 anticancer drugs{vincristine(VCR), vinblastine(VBL), adriamycin(ADR), cisplatin(CPDD), etoposide(VP-16), cytosine arabinoside(ara-C), bleomycin(Bleo) and cyclophosphamide(CYC)} and the cytotoxicity-enhancing effects of (+/-)-ar-turmerone and the extracts of the crude drugs {Lithospermum erythrorhizon(LE) and Scutellaria baicalensis(SB)} on the above mentioned anticancer drugs against HL-60 and KG-1 cells among 8 anticancer drugs, VCR, VBL, ADR, and CPDD inhibited the growth of both cell lines by more than 50%, while VP-16, ara-C, Bleo, and CYC were less effective. (+/-)-ar-Turmerone had significant inhibitory effects against both cell lines, showing the ID50 values of 11.730 mcg/ml and 0.292 mcg/ml for HL-60 and KG-1 cells. respectively. But the extracts of LE and SB roots showed no significant cytotoxic effects. According to ID50 values, the cytotoxicities of VCR, VBL and ADR against HL-60 were enhanced two, eight and three times by mixing (+/-)-ar-turmerone, five, seven and three times by adding the extract of LE root, and twenty, six and three times by mixing the extract of SB root, respectively. The cytotoxicities of the above mentioned drugs against KG-1 cell were enhanced two, seven and three times by mixing (+/-)-ar-turmerone, two, three and three times by combining with the extract of LB root, and two, five and two times by adding the extract of SB root, respectively. The cytotoxicity-potentiating effects of (+/-)-ar-turmerone and the extracts of LE and SB roots against HL-60 cell were greater than KG-1 cell.
흉곽내 병변의 진단에 절단침과 세흡인침을 이용한 경피적 침생검술의 비교
이양근(Yang Keun Rhee),곽재용(Jae Yong Kwak),송정선(Jeong Sun Song),김재헌(Jae Hean Kim),유관희(Kwan Hee You),이용철(Yong Chul Lee),한영민(Young Min Han) 대한내과학회 1996 대한내과학회지 Vol.51 No.3
Objectives: Percutaneous needle biopsy in the most reliable invasive method of obtaining the confirm diagnosis of peripheral pulmonary nodule. Needle aspiration combinde with cytologic examination is being used with increasing frequency in the diagnosis of pulmonary nodules and masses. Percutaneous needle lung biopsy was done in pulmonary or mediastinal lesions using 19 Gauge fine aspiration needle or 14 Gauge cutting needle, and the results were compared with respect to diagnostic accuracy and complications. Methods: Over a periods of recent ten years. 476 patients who had localized pulmonary lesions and underwent percutaneous needle lung aspiration and biopsy using 19 Gauge fine aspiration needle or 14 Gauge cutting needle. Results: The age distribution of the patients was from sixteen to eighty-one years old. Among 467 patients, 337 patients were male and 130 patients were female. In 244 patients, sizes of the lesions were less than 4cm, in largest diameter, and in 223 patients, sizes of the lesions were larger than 4cm, but the size of the lesion was not correlated with malignancy. Diagnosic yields of the 14 Gauge cutting needle and the 19 Gauge fine needle were 88.3% and 78.3% respectively. Success rate of benigm and malignant lesion was more higher in 14 Gauge cutting needle than 19 Gauge fine aspiration needle. Common diseases were priamry lung cancer, tuberculoma, abscess in order of frequency. The complications of procedure were mild pneumothorax in ten patients, mild hemoptysis in six patients and subcutaneous emphysema in one patient. With Cutting needle and aspiration needle, complication rate was 2.73% and 2.36%, respectively. Conclusion: Percutaenous needle lung biopsy with cutting needle under the chest computed tomogram and biplane fluoroscopy had lower complication rate and high diagnostic success rate for the intrathoracic lesions
Rat Brain cDNA Library로부터 SNAP-25 유전자의 클로닝
조애리(A-Li Cho),지영미(Young-Mi Ji),유민(Min Yoo),이순철(Soon-Chul Lee),유관희(Kwan-Hee You) 대한의생명과학회 2000 Biomedical Science Letters Vol.6 No.1
SNAP-25는 presynaptic plasma membrane에 위치하는 단백질로서 synaptic vesicle의 docking과 fusion에 있어서 매우 중요한 역할을 한다. 생쥐 SNAP-25 유전자와 99%의 높은 homology를 갖고 있는 Z2 cDNA를 probe로 사용하여 쥐의 뇌 cDNA library에서 SNAP-25 유전자를 screening 하였다. 그 결과 6개의 양성 클론을 분리해 냈으며, 이들 각각을 S1, S2, S3, S4, S5, S6으로 명명하였다. 이중에서 생쥐 SNAP-25와 가장 높은 homology를 보여주고 있는 S5 클론을 선택하여 염기서열을 분석하였다. 2,100 bp의 염기서열로 구성된 쥐 SNAP-25 cDNA는 206개의 아미노산을 coding 하는 618 bp의 open reading frame을 가지고 있으며, ORF는 209~211 bp에 위치하는 AUG codon에서 시작하여 827~829 bp에 위치하는 stop codon TAA에서 끝난다. 3' untranslated region에서는 28과 19개의 CA 반복 염기서열을 보여주고 있었으며, SNAP-25 peptide sequence에서 4개의 cystein residues는 84~91에 위치하고 있었으며, amino terminus 부분에서 amphipathic α-helix를 형성하고 있는 것을 볼 수 있었다. 사람과 쥐의 SNAP-25 유전자는 88%, 생쥐와 쥐의 경우는 97%의 homology를 보여주고 있었다. 그리고 사람과 쥐의 ORF에서 염기서열은 94%, 생쥐와 쥐의 ORF에서 염기서열은 100%의 homology를 보여주고 있었으며 사람, 생쥐, 그리고 쥐의 ORF에서 아미노산 서열은 100%의 homology를 보여주고 있었다. SNAP-25 was first investigated as a neuron-specific protein preferentially expressed in CA3 pyramidal neurons of mouse hippocampus. It is a presynaptic plasma membrane protein in the nerve cell and plays an important role in the synaptic vesicle membrane docking and fusion pathway. We have recently isolated SNAP-25 cDNA from a rat brain cDNA library using a probe of Z2 cDNA. It consisted of 2,101 bp and an open reading frame (ORF) was identified between nucleotides (nt) 209 and 827. The AUG codon (nt 209~21l) was surrounded by CTACCATGG, which corresponded to the consensus sequence of ribosomal binding site. The ORF was terminated by TAA (nt 827~829) to encode a polypeptide of 206 amino acid residues. The 3'-untranslated region contained two extensive stretches of repeated (CA)28 and (CA)19 at positions 925~980 and 1645~1682. It is noteworthy that cysteine residues were clustered in the span of amino acid residues 84~91: Cys-Gly-Leu-Cys-Val-Cys-Pro-Cys. Rat SNAP-25 showed 88% and 97% identity in nucleotide sequences to that of human and mouse, respectively. Amino acid sequence of rat SNAP-25 showed 100% identity to that of mouse and human SNAP-25.