• Title/Summary/Keyword: 세포생존

Search Result 1,972, Processing Time 0.023 seconds

WAP-EPO 유전자 도입 형질전환돼지의 계대번식시 유전자 전이효율에 관한 연구

  • 이연근;박진기;민관식;성환후;임기순;양병철;김광식;김진회;장원경
    • Proceedings of the KSAR Conference
    • /
    • 2001.03a
    • /
    • pp.14-14
    • /
    • 2001
  • 본 연구는 생명공학 관련 기술개발에 의하여 신소재 물질을 생산하고 사람에게 안전하고 생리활성이 높은 고가의 의료용 단백질 생산기술을 체계화하며 형질전환 가축을 이용한 유용물질 생산에 따른 산업화와 부가가치의 극대화에 그 목적이 있다. 유즙으로 사람의 빈혈치료제인 EPO(Erythropoietin)를 분비할 수 있는 형질전환돼지를 생산하기 위하여 WAP(Whey Acidic Protein) Promoter(2.6kb) 하류에 사람의 조혈촉진유전자(EPO: 2.6kb)를 연결시켜 미세주입용 재조합 벡터(WAP-EPO : 약 7.8kb)를 구축하였다. 구축된 재조합 벡터를 1세포기 수정란에 약 2ng/ul 농도로 미세주입한 다음 외과적 방법으로 이식하였다. 이식 후 분만 모돈으로부터 생산된 자돈의 꼬리조직을 이용 게놈DNA를 추출하고 PCR 검정을 한 결과, 유즙으로 사람의 빈혈치료제를 생산할 수 있는 유전자가 도입된 형질전환돼지 $\boxDr$새롬이$\boxUl$ (♂)를 확인하였다. 또한 이렇게 꼬리조직으로부터 확인된 새롬이의 혈액과 정액을 채취, 게놈 DNA를 추출하여 외래 유전자 삽입여부를 PCR 방법으로 검정한 결과 꼬리조직과 마찬가지로 혈액 및 정액에서도 외래유전자가 삽입되었음을 확인할 수 있었다. 이렇게 생산된 형질전환돼지 $\boxDr$새롬이$\boxUl$를 이용 계대번식을 통한 F$_1$ 산자의 생산과 유전자 전이율을 확인하기 위하여 $\boxDr$새롬이$\boxUl$정액을 이용한 인공수정을 실시하였다. 인공수정은 1999년 7월 1일부터 2000년 9월 8일 까지 총 78두의 모돈을 이용하였으며, 그 중 21두의 모돈이 분만하여 인공수정에 의한 분만율은 26.92%로 나타났다.(Table Omitted) Table 1에서와 같이 새롬이 정액을 이용한 인공수정에 의해 형질전환된 F$_1$ 자돈의 형질전환율은 17.98%로 나타났으며, 32두의 형질전환자돈 중 8두(암:4두, 수:4두)는 분만과 동시에 폐사하였거나 사육중 폐사하여 현재 24두(암:12두, 수:12두)가 생존하여 F$_1$ 간 교배계획에 의해 사육되고 있다. 이 중 암컷 4두는 현재 F$_2$ 자돈 생산과 함께 유즙내로 사람 빈혈치료제의 분비 유무를 검정중에 있으며, FISH 법에 의한 외래 유전자 삽입 검정을 확인 중에 있다.

  • PDF

Effects of Isatidis Radix and it's Active Component, Tryptanthrin on the Production of Inflammatory Mediators in Lipopolysaccharide-activated Raw264.7 Cells (LPS로 활성화된 Raw264.7 cell에서 판람근 및 Tryptanthrin의 염증매개물질억제효과)

  • Park, Sook-Jahr;Lee, Jong-Rok;Jo, Mi-Jeong;Park, Sang-Mi;Buyn, Sung-Hui;Cho, Il-Je;Kim, Sang-Chan
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
    • /
    • v.24 no.1
    • /
    • pp.64-77
    • /
    • 2011
  • Objectives : 판람근(板藍根)은 십자화과에 속하는 대청(大靑) 또는 숭남의 근(根)을 건조한 것이다. 본 연구는 판람근(板藍根)이 청열해독(淸熱解毒)함에 근거하여, LPS로 활성화된 Raw264.7 cell에서 판람근(板藍根)과 그 성분중의 하나인 tryptanthrin이 염증매개물질에 미치는 효과를 살펴보고자 하였다. Methods : 세포생존율은 MTT, nitric oxide (NO)는 Griess reagent를 사용하여 측정하였으며, 각 단백질의 발현량은 Western blot 방법을 사용하였으며, cytokine 및 cyclooxygenase-2 (COX-2)는 ELISA방법을 사용하여 측정하였다. Results : LPS는 NO 및 prostaglandin E2 (PGE2)를 유의하게 상승시켰으며, 판람근(板藍根)추출물 (IRE) 및 tryptanthrin 은 이들을 유의하게 억제하였다. 그러나 판람근(板藍根)의 또 다른 성분인 indigo는 유의한 결과를 나타내지 못하였다. IRE와 tryptanthrin은 inhibitory kappa B alpha의 인산화를 억제하여, nuclear factor-${\kappa}$B (NF-${\kappa}$B)의 핵으로의 전위(轉位)를 억제하여, iNOS 및 cytokine을 억제하였다. IRE와 tryptanthrin의 PGE2 억제는, COX-2의 발현억제에서가 아니라, COX-2의 활성을 억제함에서 기인하였다. Conclusion : 이러한 결과는 판람근(板藍根)이 NF-${\kappa}$B pathway를 경유하여 iNOS의 발현 및 COX-2의 활성을 억제함을 나타내며, 이러한 판람근(板藍根)의 항염증효능은 일부 tryptanthrin의 작용에서 기인함을 시사한다.

Wound Healing of Surgical Incisions in Rhynchocypris oxycephalus (버들치 Rhynchocypris oxycephalus에서의 외과적 상처 치유)

  • 박인석;임재현;김정혜;김동수
    • Journal of Aquaculture
    • /
    • v.11 no.2
    • /
    • pp.167-172
    • /
    • 1998
  • Rhynchocypris oxycephalus was incised through the midlateral line between the dorsal and pectoral fins and the wounds were closed with absorbable synthetic suture, and here we describe histological details on the nature of second-intention wound healing, rate of healing, and the nature of granulation tissue contraction. Postoperative survivals were 95% and 99%, compared to 100% for counterpart control fish, among fish at just and 7 days after surgery, respectively. By 14~42 days after surgery, fish had no mortality. The granulation tissue was epithelialized from day 7 through day 22 postsurgery. By 28 days postsurgery, the granulation tissue had contracted the wound margins into apposition. By 35~42 days postsurgery, the granulatin tissue contained collagen fiber and the epidermis contained well-differentiated alarm substance cells ; thus, the epidermis over the wound site was histologically normal in most fish, showed that wound healing was essentially complete within 35 days of surgery.

  • PDF

Isolation and Characterization of Antitumor Agents from Xanthium strumarium L. (한국산 도꼬마리로부터 항암물질의 분리 및 특성)

  • 김현수;이인선;여수환;성림식;유대식
    • KSBB Journal
    • /
    • v.18 no.4
    • /
    • pp.324-328
    • /
    • 2003
  • In a mutagenicity test using the Salmonella typhimurium TA98 and TA100, the Xanthium strumarium L. extracts had not a mutagenicity. The extracts were assayed that antioxidative effect using a colony formation assay. The extracts showed protective effects against the cytotoxicity of H$_2$O$_2$ and increased the immunity induced by TNF and IL-1${\beta}$. The modulating effect of Xanthium strumarium L. extract on the induction of carcinogenesis by N-methyl-N'-nitro-N-nitrosoguanidin (MNNG), was investigated in Wistar rats. The GSH content was found to be reduced by MNNG treatment, but increased on adding extract. In addition the Xanthium strumarium L. extract increased p53 expression versus MNNG alone.

Development of Anti-Wrinkle Agent from Nelumbo nucifera Root Extract (연근 추출물에서 주름개선 소재의 개발)

  • Kim, Hee Jin;Kim, Tagon;Kang, Whan Yul;Baek, Hyun;Cheon, Hae Young;Kim, Bo Young;Kim, Donguk
    • Korean Chemical Engineering Research
    • /
    • v.48 no.4
    • /
    • pp.413-416
    • /
    • 2010
  • In this research, root extracts of Nelumbo nucifera was tested to see the possibility for functional cosmetic agent. 70-100% ethanol was used as solvent and nuciferin was confirmed as active component. To test cosmetic effect of root extracts of Nelumbo nucifera, safety effect(MTT assay), anti-wrinkle effect(elastase inhibition assay) and antioxidation effect(DPPH free radical scavenging assay) were measured. When 100% ethanol was used as extracting solvent, cell viability was over 80% at $100{\mu}g/ml$, which indicated that root extract of Nelumbo nucifera was suitable for cosmetic agent. Root extract of Nelumbo nucifera showed 40~50% elastase inhibition at $100{\mu}g/ml$ so that it had good anti-wrinkle characteristics. 50% antioxidation capacity($FSC_{50}$) was $5.0{\sim}38{\mu}g/ml$ and root extract of Nelumbo nucifera showed excellent antioxidation effect. From the research, root extracts of Nelumbo nucifera showed strong possibility for anti-wrinkle functional cosmetic agent.

Micropropagation of Cypripedium guttatum and Cypripedium macranthos (조직배양기술을 통한 털복주머니란과 복주머니란 기내증식)

  • Bae, Kee-Hwa;Choi, Yong-Eui
    • Proceedings of the Plant Resources Society of Korea Conference
    • /
    • 2010.05a
    • /
    • pp.13-13
    • /
    • 2010
  • 복주머니난(Cypripedium)속 식물은 우리나라에 광릉요강꽃을 비롯하여 털복주머니난, 흰털복주머니란, 복주머니란, 노랑복주머니란등 5종이 분포하는 것으로 알려져 있다. 광릉요강꽃과 털복주머니란 두 종은 환경부에서 지정한 멸종위기 식물 1급과 2급에 지정되어 보호를 받고 있고 나머지 3종은 제도적으로도 보호를 받지 못하는 실정이다. 본 연구에서는 충분히 성숙한 털복주머니난과 복주머니난의 종자에 NaOCl처리를 하여 발아율을 향상시킬수 있었는데 이러한 전처리가 발아에 미치는 원인에 대한 연구를 실시하였다. 털복주머니란의 무균적인 종자발아는 1.0% NaOCl 처리구에서 70% 이상의 종자발아율을 보였으며, POM배지가 MS배지보다 신초분화가 양호했다. GA3와 활성탄(Activated charcoal)의 혼합첨가는 신초증식에 효과적이었다. 신초분화 후 저온처리는 신초의 증식 율을 증가시켰다. 한편 NaOCl 농도(0, 1, 3, 5, 10%)와 NaOCl 처리시간(0, 5, 15, 30, 45, 60분)에 따라서 복주머니란의 종자발아를 확인한 결과, NaOCl 1%를 30분간 처리하였을 때 가장 높은 발아율을 나타냈다. NaOCl을 처리하여 종자의 종피상태를 SEM과 TEM으로 관찰한 결과 NaOCl의 처리는 종피 세포벽의 부분적 해리 및 작은 구멍을 만들게 하였는데 이러한 종피의 물리화학적 변화가 물과 양분의 이동을 원활히 하여 종자의 발아를 촉진하는 것으로 사료된다. 복주머니난의 신초분화에 미치는 casein과 활성탄의 효과를 알아본 결과 casein 200 mg/L와 활성탄 200 mg/L를 혼합 첨가한 실험구에서 가장 높은 신초분화율을 보였다. 토양순화 후 생존률은 극히 저조했으며 30 개체중에 5 개체만이 다음해 어린동아를 싹틔우는 것을 확인 하였다. 본 결과들을 종합하여 보면 멸종위기식물, 특히 털복주머니란과 복주머니란의 조직배양을 통해서 서식지외 보존방안(기내증식)에 관해 가능성을 제시하였다고 생각되어 진다.

  • PDF

Studies on Antioxidant and Whitening Activities of Salix gracilistyla Extracts (갯버들 추출물의 항산화 및 미백활성 연구)

  • Jeong, Yong-Un;Park, Young-Jin
    • Journal of the Society of Cosmetic Scientists of Korea
    • /
    • v.44 no.3
    • /
    • pp.317-325
    • /
    • 2018
  • This study was carried out to evaluate the antioxidant and whitening activities of Salix gracilistyla extracts. The total polyphenol contents of the extracts were 142.60-151.95 mg GAE/g and total flavonoid contents were 83.43-92.60 mg CE/g. In 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay, the 30% methanol extract showed the highest antioxidant activity ($IC_{50}$; $19.68{\mu}g/mL$). Tyrosinase inhibitory activity was similar in all four solvent extracts, and the highest inhibitory activity (35.18%) was obtained at a concentration of $200{\mu}g/mL$ of 30% ethanol extract. In addition, methanol extracts did not affect cell viability at all treatment concentrations and were found to significantly reduce the melanin content of B16F10 cells. As a result, it is considered that Salix gracilistyla extracts can be used as an effective cosmetic ingredient having antioxidant and whitening activity.

Proper Condition of Centrifugation for the Fat Cell Viability in the Autologous Fat Injection (자가지방주입에서 지방세포의 생존을 위한 적절한 원심분리 조건)

  • Jang, Kyung Min;Kim, Jong Yeop;Yang, Jung Duk;Chung, Ho Yun;Park, Jae Woo;Cho, Byung Chae
    • Archives of Plastic Surgery
    • /
    • v.33 no.4
    • /
    • pp.423-426
    • /
    • 2006
  • Purpose: In the autologous fat injection, the centrifugation is useful for the refinement of harvested fat. As it can be an injury to the fat cell, we studied the fat cell viability with the change of centrifugation velocity and centrifugation time in order to get the limits of centrifugation velocity and centrifugation time. Methods: We used the Colman System in 8 patients. We handled the control group with no centrifugation, group I with the centrifugation with 1500rpm for 1 minute, group II with 1500 rpm for 3 minutes, group III with 1500rpm for 5 minutes, group IV with 3000rpm for 1 minute, group V with 3000rpm for 3 minutes, group VI with 3000rpm for 5 minutes, group VII with 5000rpm for 1 minute, group VIII with 5000rpm for 3 minutes, group IX with 5000rpm for 5 minutes. We used the collagenase to separate the fat tissue. We had evaluated the fat cell viability by checking survival cell counts. Results: There was no significance in group I, II, IV, V, but there was significant difference in group III, VI, VII, VIII, IX. Conclusion: The centrifugation with 3000rpm for 3 minutes is recommendable.

Sewage Treatment Using Electric Field Pretreatment and Metal Membrane Microfiltration Process (전기장 전처리 및 금속 정밀여과 공정을 이용한 하수처리)

  • Jung, Jongtae;Kim, Jongoh
    • Journal of the Korean GEO-environmental Society
    • /
    • v.8 no.5
    • /
    • pp.15-21
    • /
    • 2007
  • This study was investigated to evaluate the effect of electric fields application for the reduction of membrane fouling, the inactivation of microorganisms and the enhancement of particle coagulation in microfiltration/coagulation. Cumulative permeate in the case with electric field treatment showed high value compared to the case without electric field. Particularly, the increase of HRT (Hydraulic retention time) showed the increase of cumulative permeate as well as the decrease of total resistance. By electric field treatment, the amount of small particle size less than 100 ${\mu}m$ is increased and the survival fraction is reduced by the increase of electric field intensity. This may be attributed to the cleavage of particle or cytological destruction resulting from the inactivation of microorganism. Coagulant dosage of PAC compared with no electric field application could also be saved about 75% for the 95% of turbidity removal at 10 kV/cm.

  • PDF

OVERALL FIVE-YEAR SURVIVAL RATE IN SQUAMOUS CELL CARCINOMA OF ORAL CAVITY (한국인에서 구강 편평세포암종의 5년 생존율)

  • Oh, Min-Seok;Kang, Sang-Hoon;Kim, Hyung-Jun;Zhenglin, Zhao;Ryu, Jae-In;Nam, Woong;Cha, In-Ho
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
    • /
    • v.35 no.2
    • /
    • pp.83-88
    • /
    • 2009
  • The purpose of this epidemiologic study was to provide clinically useful information on the fundamentals for both the diagnosis and treatment planning of oral squamous cell carcinoma, which comprises $80{\sim}90%$ of all oral cancers. One hundred and forty two patients diagnosed with oral squamous cell carcinoma were selected from a total of 220 patients with oral malignancies. The patients' medical and follow-up records were reviewed and their survival was traced. The highest occurrence rate was observed in those aged between 60 and 69 years. The tongue was the most common primary site(31.7%) for oral squamous cell carcinoma. The survival rate was calculated using the Kaplan-Meier method. The overall five-year survival rate of oral squamous cell carcinoma patients was 66.90%. The 5-year survival rate according to stage was 85.82% for stage I, and 49.98% for stage IV. The five-year survival rate according to the originating site was 91.67% for the retromolar trigone, 75.30% for the tongue, and 62.41% for the maxillary gingiva. In terms of cell differentiation, the majority(58.5%) was the well-differentiated type, which had a 5-year survival rate of 70.62%.