• 제목/요약/키워드: Expression rate

검색결과 1,971건 처리시간 0.027초

Agrobacterium tumefaciens vector를 이용(利用)한 양황철의 형질전환(形質轉換) (Transformation of Populus nigra × P. maximowiczii Using Agrobacterium tumefaciens vectors)

  • 손석규;현정오
    • 한국산림과학회지
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    • 제87권2호
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    • pp.164-172
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    • 1998
  • Promoter가 없는 외래유전자를 이용하여 형질전환(形質轉換)시키고자 할 때의 최적조건(最適條件) 및 효율적인 형질전환체(形質轉換體)의 검정방법을 구명(究明)하고자 산림청(山林廳) 임목육종연구소(林木育種硏究所)에 식재된 잡종 포플러 양황철 62-9클론을 사용하여 재분화를 유도하고, 기내(器內) 형질전환실험(形質轉換實驗)을 실시하였다. 기내(器內)에서 식물체 재분화 유도를 위해서는 배지조성과 호르몬 농도의 조합이 가장 영향을 많이 끼치는 요인이었다. 여러 조합으로 엽조직 ($5{\times}5mm$ leaf strip)을 시료로 하여 MS 기본배지에 0, 0.01, 0.1, 0.5, $0.1mg/{\ell}$ NAA, 0.2, 0.5, 1.0, $2.0mg/{\ell}$ BAP를 혼합 처리하여 배양한 결과, $0.1mg/{\ell}$ NAA, $0.5mg/{\ell}$ BAP를 첨가한 호르몬 조합에서 즐기 분화율 및 explant당 분화된 부정줄기수가 상대적인 비율 94%(11.4개)로서 가장 높았다. 따라서 형질전환(形質轉換)된 엽조직(葉組織)을 재분화 시킬 때 이 조건을 줄기분화유도 배지 (SIM ; shoot-inducing medium)로 사용하였다. 선발유전자에 의한 항생제(抗生濟)의 선발농도(選拔濃度)를 알기 위해 pBI121로 형질전환(形質轉換)실험을 한 결과, cocultivation한 후 $100mg/{\ell}$ Km(kanamycin) 또는 $60mg/{\ell}$ G418(geneticin)이 첨가된 SIM 3 배지에서 2주가 경과하면서 육안으로 형질전환(形質轉換)된 부정아를 관찰할 수 있었고 이 부정아를 SIM 3로 옮긴지 6주 후에는 0.5-1cm 크기의 부정줄기로 자랐다. 그러나 대조(對照) 엽조직(葉組織)은 부정아형성이 거의 되지 않아, 선발유전자의 항생제(抗生濟) 선발농도(選拔濃度)는 이 조건이 최적임을 알 수 있었다. 또한 형질전환체(形質轉換體)의 재분화율을 높이기 위해 5-azacytidine을 처리한 결과, 항생제(抗生濟) 선발배지하에서 재분회율을 5.7%에서 26.7%까지 높일 수 있었다. Fluorometric 및 histochemical assay 방법으로 GUS 유전자의 활성(活生)을 검정한 결과 vector system간에 형질전환율(形質轉換率)이 서로 다름을 확인할 수 있었다. LBA4404/pBI121을 vector로 사용한 양황철의 기내(器內) 형질전환(形質轉換) 실험에서는 낮은 形짧훌훌換率(5.7%)을 보였다. 그러나 보다 응양성(應梁性)이 높은 super virulence 유전자를 포함하는 pEHA101을 helper plasmid로 하여 실험한 결과 형질전환율(形質轉換率)이 35.9%로 pAL4404보다 훨씬 효과적인 것으로 나타났다. 따라서 promoter가 없는 외래유전자를 이용하여 promoter tagging을 하고자 할 때 형질전환(形質轉換)실험에는 helper plasmid로써 pEHA101을 이용하는 것이 적합한 것으로 판명되었다.

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인간 제대혈액에서 유래된 중간엽 줄기세포의 신경 및 콜린성 분화 (Neural and Cholinergic Differentiation of Mesenchymal Stem Cells Derived from the Human Umbilical Cord Blood)

  • 감경윤;강지혜;도병록;김해권;강성구
    • 한국발생생물학회지:발생과생식
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    • 제11권3호
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    • pp.235-243
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    • 2007
  • 인간 제대혈 세포는 조혈모세포, 중간엽 줄기세포와내피전구세포를 풍부하게 포함하고 있다. 인간 제대혈 속의 중간엽 줄기세포는 조혈모세포와는 달리 다능성 줄기세포이며 신경세포로 분화할 수 있는 잠재성을 가지고 있다. 본 연구에서는 세포배양을 통해 제대혈의 중간엽 줄기세포를 신경세포와 콜린성 신경세포로 분화를 유도하였다. 중간엽 줄기세포를 신경세포로 분화시키기 위해 배양액에 dimethyl sulphoxide(DMSO)와 butylated hydroxyanisole(BHA)를 첨가하여 유도하였으며 basic fibroblast growth factor(bFGF), retinoic acid(RA), sonic hedgehog(Shh)를 처리하여 콜린성 신경세포로 분화시켰다. DMSO와 BHA에 처리된 중간엽 줄기세포가 빠르게 신경세포 모양으로 분화하는 것을 관찰하였으며, 이것은 면역조직학적 염색에서 신경세포 특이 표지인 $\beta$-tubulin III, 별아교세포에 대한 특이 표지인 GFAP, 희돌기아교세포에 대한 특이 표지인 Gal-C에 대해 양성반응을 나타내었고, 그 비율은 각각 $32.3{\pm}2.9%$, $11.0{\pm}0.9%,\;9.4{\pm}1.0%$였다. RT-PCR 분석에서 배양 단계에 따라 신경세포에 특이적인 표지 인자가 발현됨을 통해, 중간엽 줄기세포가 신경세포로 분화됨을 확인하였다. 또한, 중간엽 줄기세포에 bFGF, RA, Shh를 처리하여 콜린성 신경세포로 분화시켰을 때, 전체 중간엽 세포 중 $31.3{\pm}3.2%$가 신경세포 특이 표지인 $\beta$-tubulin III에 양성반응을 보였으며 이들 세포 중 $70.0{\pm}7.8%$가 콜린성 신경 특이 표지인 ChAT에 양성반응을 보였고, 이것은 Woodbury 방법에 의한 신경분화의 경우보다 3배 가량 높은 비율로 콜린성 신경의 분화를 유도한 것이다. 이러한 실험 결과들은 인간 제대혈의 중간엽 줄기세포가 콜린성 신경세포로 분화가 가능하고 이러한 잠재성을 가진 제대혈 중간엽 줄기세포는 퇴행성 신경질환에 대한 세포 치료제로서 가능성을 제시한다.

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비/비인강 비호즈킨 림프종의 임상양상과 예후 인자 : 면역 표현형의 임상적 의의 (Clinical Features and Prognostic Factors in 41 Patients of Primary Nasal/Nasopharyngeal Non-Hodgkin's Lymphoma: Prognostic Significance of Immunophenotype)

  • 박순서;서철원;박종범;박지운;임소덕;허주령;남순열;김상윤;이호규;장혜숙;김태원;이제환;김성배
    • 대한두경부종양학회지
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    • 제15권2호
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    • pp.149-155
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    • 1999
  • Objectives: To study the clinical features of the primary nasal/nasopharyngeal non-Hodgkin's lymphomas and to evaluate the implication of immunophenotyping as a prognostic factor. Patients and Methods: From January 1990 to December 1997,41 patients(median age, 41 years) of primary nasal/nasopharyngeal non-Hodgkin's lymphoma were studied. The clinical records and paraffin-embedded tissue blocks were reviewed retrospectively. The histologic features, immunophenotypic findings(pan-T, pan-B, CD3, CD56) and Epstein-Barr virus in situ hybridizatios were examined. The prognostic factors for clinical outcome were evaluated in these patients. According to Ann-Arbor system, there were 30 patiets(73%) with stage IE, 4(10%) with stage IIE, 3(7%) with stage IIIE, 4(10%) with stage IVE lymphoma. Among the patients with stage IE/IIE, 4 patients received local radiation alone, 4 received chemotherapy alone, 25 received combination chemotherapy and radiotherapy and 1 refused treatment. The patients with stage IIIE/IVE were given combination chemotherapy and radiotherapy. Results: Immunophenotyping were performed in 40 patients and staining results were as follows: 3(7%) patients with B cell, 17(42%) with T cell, 18(44%) with NK/T cell(CD56 positive), and two patients with unclassifiable result. Epstein-Barr(EB) virus in situ hybridization were performed in 28 patients and 23(82%) patients had positive EBV-encoded RNAs(EBERs). 21(55%) patients achieved a complete remission. There was no difference in complete remission between radiation alone and combination therapy. With median follow-up of 30 months, 5-years disease free survival of complete responders was 60% and 5-years overall survival rate was 36%. Multivariate analysis showed that better overall survival was related with absence of B symptoms, ECOG performance${\leq}1$ and non-NK cells. Conclusion: Most of all cases were positive for EBER. Since NK/T phenotype carried the worst prognosis, analysis for CD56 expression should be done. Further prospective studies were warranted to evaluate the role of chemotherapy in stage IE/IIE.

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The effect of two Terpenoids, Ursolic acid and Oleanolic acid on epidermal permeability barrier and simultaneously on dermal functions

  • Lim Suk Won;Jung Sung Won;Ahn Sung Ku;Kim Bora;Ryoo Hee Chang;Lee Seung Hun
    • 대한화장품학회지
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    • 제29권2호
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    • pp.205-232
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    • 2003
  • Ursolic acid (UA) and Oleanolic acid (ONA), known as urson, micromerol and malol, are pentacyclic triterpenoid compounds which naturally occur in a large number of vegetarian foods, medicinal herbs, and plants. They may occur in their free acid form or as aglycones for triterpenoid saponins, which are comprised of a triterpenoid aglycone, linked to one or more sugar moieties. Therefore UA and ONA are similar in pharmacological activity. Lately scientific research, which led to the identification of UA and ONA, revealed that several pharmacological effects, such as antitumor, hepato-protective, anti-inflammatory, anticarcinogenic, antimicrobial, and anti-hyperlipidemic could be attributed to UA and ONA. Here, we introduced the effect of UA and ONA on acutely barrier disrupted and normal hairless mouse skin. To evaluate the effects of UA and ONA on epidermal permeability barrier recovery, both flanks of 8-12 week-old hairless mice were topically treated with either 0.01-0.1 mg/ml UA or 0.1-1 mg/ml ONA after tape stripping, and TEWL (Transepidermal water loss) was measured . The recovery rate increased in those UA or ONA treated groups (0.1 mg/ml UA and 0.5 mg/ml ONA) at 6 h more than $20\%$ compared to vehicle treated group (p<0.05). Here, we introduced the effects of UA and ONA on acute barrier disruption and normal epidermal permeability barrier function. For verifying the effects of UA and ONA on normal epidermal barrier, hydration and TEWL were measured for 1 and 3 weeks after UA and ONA applications (2mg/ml per day). We also investigated the features of epidermis and dermis using electron microscopy (EM) and light microscopy (LM). Both samples increased hydration compared to vehicle group from f week without TEWL alteration (p<0.005). EM examination using RuO4 and OsO4 fixation revealed that secretion and numbers of lamellar bodies and complete formation of lipid bilayers were most prominent $(ONA{\geq}UA>Vehicle)$. LM finding showed that thickness of stratum corneum (SC) was slightly increased and especially epidermal thickening and flattening was observed (UA>ONA>Veh). We also observed that UA and ONA stimulate epidermal keratinocyte differentiation via $PPAR\;\alpha$. Protein expression of involucrin, loricrin, and filaggrin increased at least 2 and 3 fold in HaCaT cells treated with either $ONA\;(10{\mu}M)$ or UA $(10{\mu}M)$ for 24h respectively. This result suggested that the UA and ONA can improve epidermal permeability barrier function and induce the epidermal keratinocyte differentiation via $PPAR\;{\alpha}$. Using Masson-trichrome and elastic fiber staining, we observed collagen thickening and elastic fiber elongation by UA and ONA treatments. In vitro results of collagen and elastin synthesis and elastase inhibitory activity measurements were also confirmed in vivo findings. These data suggested that the effects of UA and ONA related to not only epidermal permeability barrier functions but also dermal collagen and elastic fiber synthesis. Taken together, UA and ONA can be relevant candidates to improve epidermal and dermal functions and pertinent agents for cosmeseutical applications.

족삼리(足三里) 사과락약침(絲瓜絡藥鍼)이 생쥐의 Collagen-induccd Arthritis에 미치는 영향 (A Study on the Effect of Luffae Fructus Retinervus Herbal-Acupuncture at ST36 on Collagen-induced Arthritis in Mice)

  • 채충헌;최선미;임윤경
    • Journal of Acupuncture Research
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    • 제22권1호
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    • pp.131-144
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    • 2005
  • 족삼리(足三里) 사과락약침(絲瓜絡藥鍼)이 DBA/lJ mouse의 collagen-induced arthritis 미치는 영향(影響)을 관찰(觀察) 한 결과(結果), 다음과 같은 결론(結論)을 얻었다. 1. 세포독성(細胞毒性) 측정결과(測定結果), 사과락약침액(絲瓜絡藥鍼液) 1%의 농도(濃度)에서 가장 높은 세포생존률(細胞生存率)이 관찰(觀察)되었다. 2. In vitro screening 결과(結果), 사과락약광액(絲瓜絡藥鑛液) 1%의 농도(濃度)에서 CIA mouse 관절활막세포(關節滑膜細胞)의 TNF-${\alpha}$발현(發現)이 유의(有意)하게 감소(減少)하였다. 3. 족삼리(足三里) 사과락약침(絲瓜絡藥鍼)에 의하여 관절염(關節炎) 발병률(發病率)이 감소(減少)하였다. 4. 족삼리(足三里) 사과락약침(絲瓜絡藥鍼)에 의하여 CIA mouse의 각장(脚臟)무게가 감소(減少)하였다. 5. 족삼리(足三里) 사과락약침(絲瓜絡藥鍼)에 의하여 CIA mouse의 혈청(血淸) IL-6, INF-${\gamma}$, TNF-${\alpha}$, IgG, IgM 및 Anti-collagen Ⅱ가 유의(有意)하게 감소(減少)하였다. 6. 조직학적(組織學的) 검사(檢査) 결과(結果), 족삼리(足三里) 사과락약침(絲瓜絡藥鍼)에 의하여 CIA mouse 관절(關節)의 연골파괴(軟骨破壞)와 활막증식(滑膜增殖)이 감소(減少)되고 교원질섬유(膠原質纖維)가 정상군(正常群)과 유사(類似)하게 유지(維持)되었다. 7. 임파절내(淋巴節內) CD3e+와 CD19+세포(細胞) 비율(比率), CD4+와 CD8+세포(細胞) 비율(比率)은 사과락약광치료군(絲瓜絡藥鑛治療群)에서 정상군(正常群)과 유사(類似)하게 유지(維持)되었다. 8. 임파절내(淋巴節內) CD69+/CD3e+세포(細胞) 및 CD11a+/CD19+세포(細胞)와 관절내(關節內) CD11b+/Gr-1+세포(細胞)는 사과락약침치료군(絲瓜絡藥鍼治療群)에서 대조군(對照群)에 비하여 유의(有意) 하게 감소(減少)하였다.

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노령출산에 관한 연구 (A Study on Childbrith in Late Maternal Age)

  • 한예영
    • 한국보건간호학회지
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    • 제8권2호
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    • pp.101-113
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    • 1994
  • Maternity means all the women who are capable to conceive. In the aspect of health and medical care. however. it means the women who are now in pregnancy or have already given birth to a baby or are in a period of being recoverd from physiological changes occurred by pregnancy. According to the rapidly changing social structure. both the Quality and Quantity of the capacity of childbirth experienced by women are changing. Our society. having established a great economical growth by virtue of the highly developing and growing industrialization and urbanization. stimulates the women's advance into society and thereby increasing the number of employed women. When the women's participation in society is increased. their age of marriage is also affected. Which means there are a decrease of the capacity of childbirth in terms of quantity and a trend for women to have less children and to deliver a baby in their old age in terms of quality. On the contrary. since the number of multipara who want to have a baby in their old age is increasing. as a counter functional effect to the political project of decrease of a birth rate. concern has been focussed on childbirth in old age in the present study. And also such kind of the childbirth may be danger to the health of both mother and baby. Therefore the present study intended to provide some basic data of health education in the part of the health management of both mother and baby in the general hospital. based on understanding the realities of childbirth in. old age and things related to them. To achieve such a purpose of the present study. an analytical study by means of SPSS. was done using the data of 269 clinical records on both the newborn .babies and their mothers who had been supported by public general hospitals located in Seoul for 3 years from Jan. 1. 1991 to Dec. 31. 1993. Some significant results from the analytic study are as follows: 1. It appeared that the average age of normal. natural delivery was 33.8 years old and the average age of delivery through the cesarean operation was 35.4 years old. 2. It appeared that danger factors to childbirth women were types of the delivery and placental extrusion and danger factors to newborn babies are not so outstanding. 3. It appeared that the variables of the childbirth capacity which showed a significant difference according to each age group of women were the number of pregnancy. number of still birth, and number of existing children. That is. the age group of 'more than 35 years' had more frequency of experience In all 3 variables than the age group of 'less than 35 years'. 4. It appeared that the variables of the childbrith capacity which showed a significant difference a according to the sex of a newborn baby were number of pregnancy, number of still birth, and number of existing children. That is, the age group of 'more than 35 years' had more frequency of experience in all 3 variables than the age group of 'less than 35 years'. 5. It appeared that the health index of newborn babies which showed a significant difference was only 5 minute APGAR. That is, the health index 9.46 in the age group of 'more than 35 years' was less than an index of 9.72 in the age group of 'less than 35 years'. 6. Since a counter correlation of -0.10, as Pearson Correlation Coefficient, was showed between the age of childbirth mothers and the weight of newborn babies, it indicated that the higher age of childbirth woman, the lesser the weight of newborn baby. 7. It appeared that the number of women who had confirmed the sex of their baby before their delivery were 45 women, $67.2\%$ of total 67 women who had delivered a baby. and the expected sex by women in childbed was male with $73.1\%$ of total childbirth women expecting male birth and with their expression of feeling of female delivery. very regretful' by $39.3\%$ of total childbirth women. The results as shown above may indicate that instead of the possibility of danger to both the mother delivering a baby in old age and the baby delivered, the expectation of getting a son motivates childbirth in old age. As a conclusion, in a dimension of general hospital as well as national reform. it is required that a program of health education for childbirth in late maternal age have to be developed in the part of the health management of both mother and baby in the near future.

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HCT116 대장암세포에서 AKT/mTOR/GSK-3β 신호경로 조절을 통한 벌 사상자 추출물(CME)의 apoptosis 및 cell cycle arrest 효과 (Apoptotic Effects and Cell Cycle Arrest Effects of Extracts from Cnidium monnieri (L.) Cusson through Regulating Akt/mTOR/GSK-3β Signaling Pathways in HCT116 Colon Cancer Cells)

  • 임은경;김근태;김보민;김은지;하성호;김상용;김영민
    • 생명과학회지
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    • 제26권6호
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    • pp.663-672
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    • 2016
  • 벌 사상자[Cnidium monnieri (L.) Cusson]는 중국과 한국에 분포하는 일년생 초본으로, 화농성피부염 및 여성의 생식기 질환의 치료에 널리 사용되고 있다. 이 외에도 면역기능개선과 천식 등에 대한 효과는 보고된 바 있으나 아직까지 암과 관련된 연구는 많이 이루어지지 않았다. 이에 따라 본 연구에서는 인간 대장암 세포인 HCT116 세포주에서 벌 사상자 에탄올 추출물(CME)의 apoptosis 및 세포주기정지 유도 효과에 대하여 알아보고자 하였으며, 이러한 효과가 AKT/mTOR/GSK-3β 신호경로의 조절을 통하여 이루어지는지 확인하고자 하였다. MTT assay와 LDH assay 결과, 벌 사상자 에탄올 추출물에 의하여 HCT116 세포의 세포생존율이 감소하였으며, 세포독성효과가 나타났다. 또한 벌 사상자 에탄올 추출물의 농도의존적으로 apoptotic body의 수와 apoptosis 비율이 증가하였으며, G1기에서 세포주기정지 유도 효과가 관찰되었다. 세포의 성장과 증식 및 분열에 관련된 단백질인 Akt는 mTOR, p53, GSK-3β와 같은 신호단백질들의 발현을 조절하는 것으로 보고되었다. 벌 사상자 에탄올 추출물을 처리하였을 때, Akt와 mTOR 단백질의 인산화가 저해되었으며, 이에 따라 하위 신호조절 단백질인 GSK-3β, Bcl-2 family, Caspase-3, PARP의 발현이 조절되었다. 또한 Akt와 GSK-3β, mTOR 저해제 처리를 통하여 CME에 의한 apoptosis 효과가 AKT/mTOR/GSK-3β 신호경로를 통하여 이루어지는 것을 확인하였다. 결론적으로, 본 연구를 통하여HCT116 대장암 세포주에서 벌 사상자 에탄올 추출물이 암세포의 apoptosis 및 세포주기정지 유도에 효과적임을 확인하였다.

식물세포배양(植物細胞培養)에 의한 제초제저항성(除草劑抵抗性) 변종선발(變種選拔) (Selection of Herbicide Tolerant Variant Through Cell Culture)

  • 김순철;정근식
    • 한국잡초학회지
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    • 제7권1호
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    • pp.90-97
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    • 1987
  • 세포배양방법(細胞培養方法)을 이용(利用)하여 제초제저항성세포(除草劑抵抗性細胞) 나아가서는 저항성(抵抗性) 식물체(植物體)를 얻기 위해 수종(數種)의 수도용(水稻用) 제초제(除草劑)를 사용(使用)하여 1986년(年)~'87년(年) 영남(領南) 작물(作物) 시험장(試驗場) 생명공학실험실(生命工學實驗室)에서 실험(實驗)을 실시(實施)하였던 결과(結果)는 다음과 같다. 1. 세포배양(細胞培養)에 필요(必要)한 Callus는 현미(玄米)를 통(通)하여 얻을 수 있으며, 치상당시(置床當時) 현미(玄米)의 숙도(熟度)가 Callus 유기(誘起) 속도(速度)와 유기율(誘起率)에 크게 영향을 미쳤는데, 출수후(出穗後) 10~15일(日)된 종자(種子)가 Callus 유기속도(誘起速度)가 빠르면서 유기율(誘起率)도 높았다. 또한 출수후(出穗後) 일수(日數)가 경과(經過)함에 따라 직선적(直線的)으로 그 효과(效果)가 저하(低下)되었다. 한편 일반적(一般的)인 Callus 유기율(誘起率)은 통일형(統一型) 품종(品種)보다 일본형(日本型) 품종(品種)이 높았다. 2. 제초제(除草劑)에 대(對)한 저항성(抵抗性) Callus 출현율(出現率)은 제초제종류(除草劑種類)와 대상품종(對象品種) 또는 식물(植物)의 종(種)에 따라 차이(差異)를 보였다. 낙동(洛東)벼의 경우 제(第) 1 차배양(次培養)에서 CGA 142464 제초제(除草劑)가 함유(含有)된 배지(培地에)서 가장 높은 저항성(抵抗性) Callus 출현율(出現率)(46.3%)을 보였고 다음으로는 NC-311, 11.6%, butachlor 7.5%, 2,4-D 2.1% Quinclorac 0.89%, Propani1 0.25%의 순(順)이었다. 또한 이들 저항성(抵抗性) Callus 출현율(出現率)은 직대배양(織代培養) 회수(回數)가 진전(進展)되면서 급속도(急速度)로 증가(增加)될 뿐 아니라 본래(本來)의 농도(濃度)보다 10배(倍), 100배(倍)나 높은 농도(濃度)에서도 상당(相當)한 Callus가 저항성(抵抗性)을 보였다. 3. 제초제(除草劑) 저항성(抵抗性) Callus의 식물체(植物體) 재분화능력(再分化能力)은 배지(培地) 종류(種類)와 제초제(除草劑) 종류(種類)에 따라 달랐으나 대체로 $N_6-Y_1$의 기본배지(基本培地)에 2,4-D와 kinetine이 각각(各各) 0.2mg과 2 mg (l당(當))이 함유(含有)된 배지(培地)에서 유아부(幼芽部)와 유근부(幼根部)가 다같이 분화(分化)가 되었을 뿐 그외 배지(培地)에서는 뿌리만 분화되거나, Callus만 증식(增植)되었다. 한편 유아(幼芽)와 유근(幼根)이 함께 분화(分化)된 것은 정상적(正常的)인 식물체(植物體)로 생장(生長)되지 못하였다.

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인간 착상 과정에 자궁내막과 배아의 역할 (The Role of the Endometrium and Embryo in Human Implantation)

  • 지병철
    • 한국발생생물학회지:발생과생식
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    • 제13권1호
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    • pp.1-11
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    • 2009
  • 착상은 배아측과 모체측의 내분비적, 측분비적 및 자가분비적 인자들의 상호 작용에 의하여 조절된다. 착상의 적기는 $2{\sim}4$일로서 이 특수한 시간대 안에서 signaling, appositioning, attachment 및 invasion을 통하여 순차적으로 배아의 착상이 이루어지는데, 이는 자궁내막과 배아로부터 여러 사이토카인, 성장인자, 부착인자와 같은 다수의 생화학 인자의 생성과 분비를 포함하며 이로 인하여 수용적인 자궁내막이 형성된다. LIF, CSF-1, IL-1과 같은 사이토카인들은 착상을 이끄는 연속적인 사건에서 중요한 역할을 수행하며 integrin, L-selectin ligands, glycodelin, mucin-1, HB-EGF, pinopodes는 appositioning과 attachment에 관여한다. 배아 또한 사이토카인과 성장인자 및 LIF, CSF-1, IGF, HB-EGF에 대한 수용체들을 분비하여 능동적으로 대처한다. 자궁내막의 수용성을 평가하고 자연주기 또는 보조생식술을 이용한 임신의 예후를 예측하는데, 이와 같은 인자들이 유용한지는 앞으로 더 연구되어야 한다. 현재로서는 내막조직으로부터 채취한 integrins, pinopodes, glycodelin, LIF가, 자궁강내 세척액에서는 glycodelin과 LIF가 유망한 인자로 떠오르고 있다. 혈액내 마커로서는 VEGF, glycodelin, CSF가 약간의 연관성을 보여주고 있다. 이러한 인자들이 보조생식술 전후로 자궁내막의 기능과 임신의 예후를 평가하는 선별검사로 이용될 수 있는지를 증명하기 위해서는 향후 불임 여성과 대조군인 가임 여성을 대상으로 한 대규모의 연구가 필요할 것이다. 인간의 착상에서 이들 인자들의 기능을 충분히 이해해야 치료적 기법으로 연결되어 보조생식술에서도 더 높은 성공률을 이루어 낼 것으로 사료된다.

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Integrated Rotary Genetic Analysis Microsystem for Influenza A Virus Detection

  • Jung, Jae Hwan;Park, Byung Hyun;Choi, Seok Jin;Seo, Tae Seok
    • 한국진공학회:학술대회논문집
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    • 한국진공학회 2013년도 제45회 하계 정기학술대회 초록집
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    • pp.88-89
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    • 2013
  • A variety of influenza A viruses from animal hosts are continuously prevalent throughout the world which cause human epidemics resulting millions of human infections and enormous industrial and economic damages. Thus, early diagnosis of such pathogen is of paramount importance for biomedical examination and public healthcare screening. To approach this issue, here we propose a fully integrated Rotary genetic analysis system, called Rotary Genetic Analyzer, for on-site detection of influenza A viruses with high speed. The Rotary Genetic Analyzer is made up of four parts including a disposable microchip, a servo motor for precise and high rate spinning of the chip, thermal blocks for temperature control, and a miniaturized optical fluorescence detector as shown Fig. 1. A thermal block made from duralumin is integrated with a film heater at the bottom and a resistance temperature detector (RTD) in the middle. For the efficient performance of RT-PCR, three thermal blocks are placed on the Rotary stage and the temperature of each block is corresponded to the thermal cycling, namely $95^{\circ}C$ (denature), $58^{\circ}C$ (annealing), and $72^{\circ}C$ (extension). Rotary RT-PCR was performed to amplify the target gene which was monitored by an optical fluorescent detector above the extension block. A disposable microdevice (10 cm diameter) consists of a solid-phase extraction based sample pretreatment unit, bead chamber, and 4 ${\mu}L$ of the PCR chamber as shown Fig. 2. The microchip is fabricated using a patterned polycarbonate (PC) sheet with 1 mm thickness and a PC film with 130 ${\mu}m$ thickness, which layers are thermally bonded at $138^{\circ}C$ using acetone vapour. Silicatreated microglass beads with 150~212 ${\mu}L$ diameter are introduced into the sample pretreatment chambers and held in place by weir structure for construction of solid-phase extraction system. Fig. 3 shows strobed images of sequential loading of three samples. Three samples were loaded into the reservoir simultaneously (Fig. 3A), then the influenza A H3N2 viral RNA sample was loaded at 5000 RPM for 10 sec (Fig. 3B). Washing buffer was followed at 5000 RPM for 5 min (Fig. 3C), and angular frequency was decreased to 100 RPM for siphon priming of PCR cocktail to the channel as shown in Figure 3D. Finally the PCR cocktail was loaded to the bead chamber at 2000 RPM for 10 sec, and then RPM was increased up to 5000 RPM for 1 min to obtain the as much as PCR cocktail containing the RNA template (Fig. 3E). In this system, the wastes from RNA samples and washing buffer were transported to the waste chamber, which is fully filled to the chamber with precise optimization. Then, the PCR cocktail was able to transport to the PCR chamber. Fig. 3F shows the final image of the sample pretreatment. PCR cocktail containing RNA template is successfully isolated from waste. To detect the influenza A H3N2 virus, the purified RNA with PCR cocktail in the PCR chamber was amplified by using performed the RNA capture on the proposed microdevice. The fluorescence images were described in Figure 4A at the 0, 40 cycles. The fluorescence signal (40 cycle) was drastically increased confirming the influenza A H3N2 virus. The real-time profiles were successfully obtained using the optical fluorescence detector as shown in Figure 4B. The Rotary PCR and off-chip PCR were compared with same amount of influenza A H3N2 virus. The Ct value of Rotary PCR was smaller than the off-chip PCR without contamination. The whole process of the sample pretreatment and RT-PCR could be accomplished in 30 min on the fully integrated Rotary Genetic Analyzer system. We have demonstrated a fully integrated and portable Rotary Genetic Analyzer for detection of the gene expression of influenza A virus, which has 'Sample-in-answer-out' capability including sample pretreatment, rotary amplification, and optical detection. Target gene amplification was real-time monitored using the integrated Rotary Genetic Analyzer system.

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