• 제목/요약/키워드: RT-PCR analysis

검색결과 1,779건 처리시간 0.037초

별불가사리 추출물의 면역세포 활성화 효과 (Effect of Asterina pectinifera Extracts on the Activation of Immune Cells)

  • 채수연;김미정;김도순;박정은;조성기;이성태
    • 한국식품영양과학회지
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    • 제36권3호
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    • pp.269-275
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    • 2007
  • 불가사리 추출물을 이용하여 생쥐 비장에 있는 면역세포 활성화 효과에 대해 실험한 결과, 다음과 같은 결과를 얻었다. 첫째, 별불가사리 추출물 중에서 B세포와 대식세포를 활성화시키는 성분은 아세톤처리로 추출할 수 있었다. 둘째, 이 성분은 농도 의존적으로 생쥐 비장세포의 증식반응을 유도하였으며, IL-6와 $IFN-{\gamma}$의 생산을 유도하였다. 셋째, 이 성분은 B세포의 증식을 유도하였으며, 이 때 면역글로불린 IgM과 IgG의 생산도 유도하였다. 넷째, 이 성분은 대식세포주의 일산화질소 생산을 유도하였으며, 또 $TNF-{\alpha}$, GM- CSF와 IL-6의 분비를 유도하였다. 이상의 실험 결과, 본 실험에서 사용한 별불가사리 추출물에는 B세포와 대식세포 같은 면역세포의 증식과 각종 사이토카인을 생산을 유도하여, 면역반응을 조절하는 성분이 포함되어 있는 것으로 생각된다.

사람의 골수 줄기 세포로부터의 골세포 분화 과정에서 BMP-2가 미치는 영향과 그에 따른 분화 유전자의 발현 비교 연구 (THE EFFECT OF RHBMP-2 IN HUMAN BONE MARROW-DERIVED STEM CELLS AS OSTEOGENIC INDUCERS)

  • 김인숙;장옥련;조태형;이규백;박용두;노인섭;;황순정;김명진;이종호
    • Maxillofacial Plastic and Reconstructive Surgery
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    • 제27권1호
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    • pp.16-23
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    • 2005
  • It is commonly acknowledged that bone morphogenic protein (BMP-2) functions as a potential osteogenic inducer in bone formation. Recently, several papers reported that bone marrow-derived stem cell (BMSC) from human is not responsive to BMP-2 in comparison to high capacity of BMP-2 in the osteoinduction of stromal cell derived from bone marrow of rodent animals such as rat or mouse. In this study, we characterized BMSC derived from 11 years old donor for the responsiveness to rhBMP-2, dexamethasone (Dex) and 1,25-dihydroxyvitamin D (vitamin D), in order to analyze their function in the early osteogenesis. The effect of over mentioned agents was evaluated by means of assessing alkaline phosphatase (ALP) activity/staining, RT-PCR analysis and von Kossa staining. In addition, we analyzed the meaning of expressed several osteoblastic markers such as alkaline phosphatase, collagen typeI, osteopontin, bone sialoprotein and osteocalcin with relation to either differentiation or mineralization. Only in the presence of Dex, human BMSC could commit osteoblastic differentiation and matrix mineralization, and either BMP-2 or vitamin D treatment was not able to induce. But BMP-2 or Vitamin D showed potential synergy effect with Dex. ALP and bone sialoprotein were clearly expressed in response of Dex treatment compared to weak expression of osteopontin in early osteogenesis. Therefore, we expect that this study will contribute partly to elucidiating early osteogenesis mechanism in human, but variations among bone marrow donors must be considered through further study.

정전가매이진탕(正傳加味二陳湯)이 과산화지질 급여 비만 쥐의 지질강하, 항산화효과 및 염증매개물질의 생산에 미치는 영향 (Effects of Jengjengamiyijin-tang (zhengzhuanjiaweierchentang) on Lowering Lipid, Antioxidation and Production of Inflammatory Mediators Being Used Rats Fed on High Oxidized Fat)

  • 허성규;박원형;차윤엽
    • 한방재활의학과학회지
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    • 제23권4호
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    • pp.9-21
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    • 2013
  • Objectives The purpose of this study was investigating effects of Jengjengamiyijin-tang (zhengzhuanjiaweierchentang) (JGYT) on lowering lipid, antioxidation and production of inflammatory mediators being used rats fed on high oxidized fat. Methods We divided fat Sprague-Dawley rats fed on high oxidized into 4 groups. Each of 8 rats was divided into a control group and experimental groups. We fed a control group of rats a basal diet and administered normal saline (100 mg/kg, 1 time/1 day) for 4 weeks. And We fed each experimental group of rats basal diet and administered an extract of JGYT extracts (100 mg/kg, 200 mg/kg, 300 mg/kg, 1 time/1 day) for 4 weeks. At the end of the experiment, the rats were sacrificed to determine their chemical composition. We measured lipid of plasma and liver, concentration of proinflammatory cytokines, antioxidative activity and plasma tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), plasma interleukin-6 (IL-6), Apo-B, Apo-E and Leptin gene expression. Results 1. Concentration of plasma FFA, LDL-cholesterol, plasma and liver total cholesterol showed a significant decrement in JGYT groups. However, concentration of plasma HDL-cholesterol showed a significant increment in JGYT groups. 2. Concentration of plasma and liver TG, TBARS showed a significant decrement in JGYT groups. However, concentration of liver GSH-Px, SOD and CAT showed a significant increment in JGYT groups. 3. Plasma GPT activity and concentration of plasma IL-6, TNF-${\alpha}$, NO, Ceruloplasmin, ${\alpha}1$-acid glycoprotein showed a significant decrement in JGYT groups. 4. In the analysis of RT-PCR, gene expression of Apo-B and Apo-E in the JGYT groups showed a low expression than that of control group. However, the gene expression of leptin showed no difference in all the treatment groups. 5. The ratio of leptin expression per ${\beta}$-actin expression showed no significant difference among all treatment groups. However, The ratio of Apo-B and Apo-E expression per ${\beta}$-actin expression showed a significant decrement in JGYT groups. Conclusions According to this study, extract of JGYT showed a positive effect in lowering lipid, antioxidation and control of inflammatory mediators production.

맥문동탕이 호흡기 점액의 생성 및 분비에 미치는 영향 (Effect of Macmundongtang on Production and Secretion of Respiratory Mucus)

  • 성현경;민상연;김장현
    • 대한한방소아과학회지
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    • 제27권1호
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    • pp.69-81
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    • 2013
  • Objectives In this study, effects of Macmundongtang (MMT) on ATP or TNF-${\alpha}$ or PMA or EGF induced MUC5AC mucin production and gene expression from human airway epithelial cells and the increase in airway epithelial mucosubstances of rats were investigated. Materials and Methods Confluent NCI-H292 cells were pretreated for 30min in the presence of MMT and treated with ATP ($200{\mu}M$) or PMA (10 ng/ml) or EGF (25 ng/ml) or TNF-${\alpha}$ (0.2 nM) for 24hrs, to assess the effect of MMT both on ATP- or PMA- or EGF- or TNF-${\alpha}$-induced MUC5AC mucin production using enzyme-linked immunosorbent assay (ELISA) and on gene expression by the same inducers using reverse transcription-polymerase chain reaction (RT-PCR). At the same time, hypersecretion of airway mucus was induced by exposure of rats to SO2 during 3 weeks. Effect of orally-administered MMT during 2 weeks on increase in airway epithelial mucosubstances from tracheal goblet cells of rats was assesed using histopathological analysis after staining the epithelial tissue with PAS-alcian blue. Possible cytotoxicity of MMT was assessed by investigating the potential damage of kidney and liver functions by measuring serum GOT/GPT activities and serum BUN concentration of rats and the body weight gain during experiment, after administering MMT orally. Results (1) MMT did not only inhibit but also increased MUC5AC mucin productions and expression levels of MUC5AC gene from NCI-H292 cells. (2) MMT did not decrease the amount of intraepithelial mucosubstances of trachea of rats. (3) MMT did not show renal and hepatic toxicities and did not affect body weight gain of rats during experiment. Conclusions The result from the present study suggests that MMT might normalize the production and gene expression of airway mucin observed in various respiratory diseases accompanied by yin-deficiency, without in vivo toxicity to liver and kidney functions after oral administration.

담배 형질전환체를 이용한 포도 UDP-glucose flavonoid glucosyl transferase (UFGT) 유전자의 기능 분석 (Functional Analysis of a Grapevine UDP-Glucose Flavonoid Glucosyl Transferase (UFGT) Gene in Transgenic Tobacco Plants)

  • 박지연;박성출;피재호
    • 생명과학회지
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    • 제20권2호
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    • pp.292-297
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    • 2010
  • 페놀화합물인 안토시아닌은 과일, 꽃잎 등 식물 조직에서 청 혹은 적색을 나타내는 색소이다. 포도의 경우, 안토시아닌은 적포도 품종의 과피에만 축적되는데, 이것은 안토시아닌 생합성에 관여하는 효소 중에서 UDP-glucose: flavonoid 3-O-glucosyltransferase(UFGT)를 암호화하는 ufgt 유전자에 의해 조절된다고 보고된 바 있다. ufgt 유전자에 의해 안토시아닌의 축적 양상이 변하는지를 검증하기 위해, 포도로부터 ufgt cDNA를 분리한 다음, 각각 sense와 antisense 방향으로 발현하는 벡터를 제작하고, 이를 야생형 담배에 도입하였다. 담배 형질전환체를 선발하여 RT-PCR을 수행한 결과, 포도 ufgt 유전자가 sense 방향으로 들어간 snese 형질전환체에서는 야생형 담배에 비해 ufgt 전사체의 양이 증가하였고, antisense 방향으로 들어간 antisense 형질전환체에서는 전사체의 양이 도리어 감소하였다. 한편, 담배 형질전환체의 꽃을 분석한 결과, sense 형질전환체의 안토시아닌 함량은 야생형 담배에 비해 최고 44% 증가하였고, antisense 형질전환체에서는 최고 88% 감소하였다. 또한 sense 형질전환체의 꽃 색깔은 야생형 담배에 비해 더 진해졌다. 이러한 결과는 외부로부터 도입된 포도 ufgt 유전자의 발현으로 ufgt 전사체의 양이 증가하면 담배 내 안토시아닌의 함량이 높아지고 꽃 색깔이 진해진다는 것을 시사한다.

부산지역 호흡기감염증 환자로부터 분리한 아데노바이러스와 보카바이러스의 유행양상 분석 (Epidemiological Characterization of Adenovirus and Human Bocavirus Detected Acute Respiratory Patients in Busan)

  • 황수정;김남호;박동주;구평태;이미옥;진성현
    • 생명과학회지
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    • 제27권3호
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    • pp.275-282
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    • 2017
  • 부산지역 급성호흡기감염증으로 내원한 환자에서 호흡기바이러스 검사를 시행하여 아데노바이러스 및 보카바이러스의 특징적인 임상증상 및 유행양상을 알아보고자 하였다. 3,230명의 환자에서 총 1,485건(46.0%)의 호흡기바이러스가 검출되었고 이 중 아데노바이러스 257건(8.0%), 보카바이러스 68건(2.1%)을 확인하였다. 성별 발생양상은 남자가 여자보다 아데노바이러스, 보카바이러스로 인한 호흡기감염증에 취약하였으며, 모두 1~5세 연령군에서 높은 검출률을 보여 영 유아의 주요 감염요인임을 확인할 수 있었다. 주요 임상증상으로 아데노바이러스는 발열, 두통이 보카바이러스는 천명음이 통계적으로 유의하였다. 아형분석결과, 아데노바이러스는 1~6, 8형의 혈청형을 확인하였고 기존 분리주와 97% 이상의 상동성을 보였다. 연도별 유행 혈청형은 2011년 아데노바이러스 1형, 2012년 3형과 4형, 2013년에는 3형이 우점적으로 유행하였다. 아데노바이러스 3형의 경우, 대유행을 유발하는 아형으로 보인다. 보카바이러스는 모두 1형으로 나타났다.

황체호르몬 수용체의 발현이 저반응 환자군의 임신에 미치는 영향 (The effect of LH Receptor in the Pregnancy of Poor Responders)

  • 이정복;도병록;김은수;김명희;천은경;정현정;노성일;김문규;윤현수
    • Clinical and Experimental Reproductive Medicine
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    • 제31권2호
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    • pp.111-117
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    • 2004
  • Objectives: To investigate the effect of LH receptor in folliculogenesis, we confirm the expression level of LH receptor (LH-R) mRNA in human granulosa cells (GCs) and its expression levels were analyzed by comparison to embryo developmental rate and pregnancy rate. Materials and Methods: GCs were obtained at the time of oocyte retrieval from the patients undergoing IVF-ET program. The patients were divided into two groups: Group I (n=20) is poor responder (retrieved oocyte(s)$\leq$3ea), Group II (n=80) is normal responder (retrieved oocytes>3ea). After the extraction of total RNA, semiquantitative RT-PCR was performed and the expression level of LH-R mRNA was normalized by $\beta$-actin. Statistical analysis was performed by using $X^2$ test, Student's t-test and Pearson correlation. Results: In Group II, the relative values of LH-R mRNA (0.680 vs. 0.463, p<0.005) and pregnancy rate (54.7% vs. 23.1%, p<0.05) were significantly higher than in Group I. Number of retrieved oocyte(s) was gradually increased when the expression of LH-R mRNA was increased (p<0.05). But the quality of retrieved oocyte and transferred embryo were not related with the expression of LH-R mRNA. When the pregnancy rate was compared with FSH only group and FSH combined with hMG group in the ovarian stimulation protocol, FSH combined with hMG group was significantly higher than FSH only group in Group I (37.5% vs. 0%), and the expression of LH-R mRNA was significantly higher in hMG combined group than FSH only group (p<0.05). Conclusion: Expression level of LH-R mRNA has important role in ovarian function related with the response to gonadotrophin in human folliculogenesis. Furthermore these data might provide the evidence that additional use of hMG is helpful to poor responders.

Reconstructed Adeno-Associated Virus with the Extracellular Domain of Murine PD-1 Induces Antitumor Immunity

  • Elhag, Osama A.O.;Hu, Xiao-Jing;Wen-Ying, Zhang;Li, Xiong;Yuan, Yong-Ze;Deng, Ling-Feng;Liu, De-Li;Liu, Ying-Le;Hui, Geng
    • Asian Pacific Journal of Cancer Prevention
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    • 제13권8호
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    • pp.4031-4036
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    • 2012
  • Background: The negative signaling provided by interactions of the co-inhibitory molecule, programmed death-1 (PD-1), and its ligands, B7-H1 (PD-L1) and B7-DC (PD-L2), is a critical mechanism contributing to tumor evasion; blockade of this pathway has been proven to enhance cytotoxic activity and mediate antitumor therapy. Here we evaluated the anti-tumor efficacy of AAV-mediated delivery of the extracellular domain of murine PD-1 (sPD-1) to a tumor site. Material and Methods: An rAAV vector was constructed in which the expression of sPD-1, a known negative regulator of TCR signals, is driven by human cytomegalovirus immediate early promoter (CMV-P), using a triple plasmid transfection system. Tumor-bearing mice were then treated with the AAV/sPD1 construct and expression of sPD-1 in tumor tissues was determined by semi quantitative RT-PCR, and tumor weights and cytotoxic activity of splenocytes were measured. Results: Analysis of tumor homogenates revealed sPD-1 mRNA to be significantly overexpressed in rAAV/sPD-1 treated mice as compared with control levels. Its use for local gene therapy at the inoculation site of H22 hepatoma cells could inhibit tumor growth, also enhancing lysis of tumor cells by lymphocytes stimulated specifically with an antigen. In addition, PD-1 was also found expressed on the surfaces of activated CD8+ T cells. Conclusion: This study confirmed that expression of the soluble extracellular domain of PD-1 molecule could reduce tumor microenvironment inhibitory effects on T cells and enhance cytotoxicity. This suggests that it might be a potential target for development of therapies to augment T-cell responses in patients with malignancies.

Heme 산화효소 발현 유도체로서 Isoflavone-Free 검은콩 펩타이드의 항고혈압 활성 (Antihypertensive Effects of Novel Isoflavone-Free Black Soy Peptide Mixture as HO-1 Inducer)

  • 신미경;권용현;안창원;신동석;박수현;최보화;홍순선;강주희;박창신
    • 약학회지
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    • 제56권3호
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    • pp.191-197
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    • 2012
  • We previously reported that the novel isoflavone-free peptide mixture (black soybean peptide, BSP) had several beneficial effects like antiobesity and hypotriglyceridemic effect. However, there are no reports for BSP on anti-hypertensive activity. BSP induced heme oxygenase-1 (HO-1) in HUVECs, thus investigated the HO-1-induced activity in HUVECs and the anti-hypertensive effects in SHR animal model. BSP significantly induced HO-1 expression both at transcriptional and protein levels in a time- and dose-dependent manner as measured by RT-PCR and Western blot analysis, respectively. These inductions were abolished by pretreatment of N-acetyl-cystein (NAC, 1~10 mM), but not by employing Tempol, a superoxide dismutase (SOD) mimetic (1~5 mM). As expected, enzymatic activity (~2 fold) determined by bilirubin formation assay and cGMP concentration (~6 fold) were significantly increased in BSP-treated cells. Based on the numerous evidences on the beneficial effects of HO-1 and our results, we investigated in vivo effects of BSP on the antihypertensive activity. The administration of BSP (1% in drinking water) significantly decreased mean blood pressure (BP) (from $218.6{\pm}6.99$ to $190.0{\pm}3.42$ mm Hg, p<0.01). This result indicates that BSP is strong inducer of HO-1 expression, which may be triggered by oxidative stress, and has anti-hypertensive activity.

Study on CsRCI2D and CsRCI2H for improvement of abiotic stress tolerance in Camelina sativa L.

  • Lim, Hyun-Gyu;Kim, Hyun-Sung;Kim, Jung-Eun;Ahn, Sung-Ju
    • 한국작물학회:학술대회논문집
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    • 한국작물학회 2017년도 9th Asian Crop Science Association conference
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    • pp.196-196
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    • 2017
  • Oilseed crop Camelina (Camelina sativa L.) is a suitable for biodiesel production that has high adaptability under low-nutrient condition like marginal land and requires low-input cost for cultivation. Enhanced abiotic stress tolerance of Camelina is very important for oil production under the wide range of different climate. CsRCI2s (Rare Cold Inducible 2) are related proteins in various abiotic stresses that predicted to localized at plasma membrane (PM) and endoplasmic reticulum (ER). These proteins are consist of eight-family that can be divided into tail (CsRCI2D/E/F/G) and no-tail (CsRCI2A/B/E/H) type of C-terminal. However, it is still less understood the function of C-terminal tail. In this study, CsRCI2D/H genes were cloned through gateway cloning system that used pCB302-3 as destination vector. And we used agrobacterium-mediated transformation system for generation of overexpression (OX) transformants. Overexpression of target gene was confirmed using RT-PCR and segregation ratio on selection media. We analyzed physiological response in media and soil under abiotic stresses using CsRCI2D and CsRCI2H overexpression plant. To compare abiotic stresses tolerance, wild type and CsRCI2D/H OX line seeds were sown on agar plate treated with various NaCl and mannitol concentration for 7 days. In the test of growth rate under abiotic stress on media, CsRCI2H OX line showed similar to NaCl and mannitol stress. In the other hand, CsRCI2D OX line showed to be improved stress tolerance that especially increased in 200mM NaCl but was similar on mannitol media. In greenhouse, WT and CsRCI2D/H OX lines for physiological analysis and productivity under abiotic stresses were treated 100, 150, 200mM NaCl. Then it was measured various parameters such as leaf width and length, plant height, total seed weight, flower number, seed number. CsRCI2H OX line in greenhouse did not show any changes in physiological parameters but CsRCI2D OX line was improved both physiological response and productivity under NaCl stress. Among physiological parameters of CsRCI2D OX line under NaCl stress, leaf length and width were observed shorter than WT but it were slightly longer than WT in 200mM NaCl stress. Furthermore, total seed weight of CsRCI2D OX line under stress displayed to decrease than WT in normal condition, but it was gradually raised with increasing NaCl stress then more than WT relatively. These results suggested CsRCI2D might be contribute to improve abiotic stress tolerance. However, function of CsRCI2H is need to more detail study. In conclusion, overexpression of CsRCI2s family can generate various environmental stress tolerance plant and may improve crop productivity for bio-energy production.

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