• 제목/요약/키워드: PCR-ELISA

검색결과 706건 처리시간 0.027초

Clinical Significance of Expression and Amplification of the DcR3 Gene in Pancreatic Carcinomas

  • Zhou, Jian;Song, Shi-Duo;Li, De-Chun;Zhou, Jin;Zhu, Dong-Ming;Zheng, Shi-Ying
    • Asian Pacific Journal of Cancer Prevention
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    • 제13권2호
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    • pp.719-724
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    • 2012
  • This study aimed to investigate the clinical significance of expression and amplification of decoy receptor 3 (DcR3) in pancreatic carcinomas (PC). mRNA expression was detected by PQ-PCR, and amplification was determined. DcR3 protein expression was detected by immunohistochemistry and ELISA. Correlations between DcR3 expression and clinical pathological factors were analyzed. The relative amount of DcR3 in PC tissues and non-cancerous tissues showed a statistically significant difference, 21 cases displaying more than two fold DcR3 amplification, while no such amplification was found in normal pancreatic tissues. DcR3 positive cell staining was located in the cytoplasm. The positive rate of DcR3 in PC and non-cancerous tissues showed a significant difference. DcR3 mRNA expression was correlated with clinical staging, size of the tumor, lymph node metastasis and histological staging, while protein expression was correlated with clinical data like tumor size. DcR3 gene amplification only correlated with tumor size. The level of DcR3 in serum of the PC resectable group before operation was $72.2{\pm}10.2$ pg/ml, showing a significant difference compared to gallbladder carcinoma group (GC) or pancreatic benign tumor (PBT) group (P < 0.01). In conclusion, DcR3 amplification is correlated with DcR3 expression in PC tissues, especially those clinical pathological factors which reflect tumor progression. Assessment of DcR3 level in sera of PC patients may be helpful for the early diagnosis and prognostic judgement.

해독금화산 물추출물이 LPS로 유도된 대식세포의 염증반응에 미치는 영향 (Effects of Haedokgumhwa-san Water Extracts on LPS-induced Inflammatory Response in Macrophage)

  • 임재수;강옥화;서윤수;권동렬
    • 대한본초학회지
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    • 제30권5호
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    • pp.67-74
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    • 2015
  • Objectives : TheHaedokgumhwa-sanwater extract (HDKHS) is used in Korea, Japan and China as a traditional therapeutic agent to cure an infectious disease. But its study is not enough. Therefore, the present study focused on the elucidation of HDKHS to investigate the anti-inflammatory effects and to established the possible mechanisms involved in its action on LPS-stimulated immune response in murine macrophages.Methods : Inflammatory status was induced by LPS and measured by increasement of inflammatory mediators. LPS induced secretions of NO and PGE2in RAW 264.7 cells were measured using griess reagent and enzyme-linked immunosorbent assay (ELISA) kit respectively. production of IL-6 was examined using ELISA kit and expression of IL-6 mRNA was measured by RT-PCR method. To investigate the effects of HDKHS on inflammatory mediators, such as iNOS, COX-2 and MAPKs, western blot and RT-PCR were performed.Results : HDKHS significantly reduced production of NO and PGE2 which were induced by LPS. Also, activation of IL-6 was reduced both protein and mRNA levels. The expressions of inflammatory mediator include iNOS and COX-2 were decreased by pretreatment with HDKHS. futhermore The result showed HDKHS down-regulate the LPS induced phosphorylation of ERK 1/2, one of the MAPK family, which is considered as a main regulator of transmission from pathogens to nucleus of immune cells.Conclusions : Our results suggest that the anti-inflammatory properties of HDKHS may stem from the inhibition of pro-inflammatory mediators via suppression of initiation of inflammatory response by inhibiting MAPKs signaling pathways.

Effects of Mix-1 on Anti-CD40 Antibody and Recombinant IL4- Induced Cytokine Production and Immunoglobulin E in Highly Purified Mouse B Cells

  • Kim Jung Hwan;Choi Sun Mi;Lee Yong Gu;Namgoong Uk;Kim Dong Hee
    • 동의생리병리학회지
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    • 제18권6호
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    • pp.1869-1880
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    • 2004
  • In the oriental medicine, a mixture of herbs has been commonly used as important components to control allergic and inflammatory diseases. In the present study, we prepared a mixture of Dictamni Radicis Cortex(Baiksunpee), Houttuyniae Herba(Uhsungcho), and Aurantii Immaturus Fructus(Jisil) to examine its anti-allergic effects in activated mouse splenic cells and found that Mix-1 is involved in regulating levels of B cell activating factors (CD23 and CD11a), IL-1β, IL-6, IL-10, TNF-α, and 1gE as well as HRF expression. It was observed that Mix-1 did not have cytotoxic effects on mLFC. Mix-1 showed inhibition of CD23 and CD11 alpaha expression in mouse B cells, and also decreased the production of IL-6, TNF-α, and 1gE. Both RT-PCR and ELISA analyses indicated that IL-6 and TNF alpha production were regulated at the gene expression level. In contrast, IL-10 mRNA and protein levels were increased in activated B cells by Mix-1 treatment. We also found that Mix-1 inhibited B cell proliferation and inhibited histamine releasing factor(HRF) expression, suggesting its inhibitory effect on histamine secretion. These data indicated that Mix-1 has an anti-allergic effect in activated macrophages and further suggest the possible application of Mix-1 as a therapeutic agent for the treatment of allergy-related diseases.

가미패모탕이 보조 T 임파구 분화에 미치는 영향 (Effect of Gamipaemo-tang Ethanol Extract on Helper T Cell Differentiation)

  • 신재호;고은정;홍무창;정승기;신민규;배현수
    • 동의생리병리학회지
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    • 제18권5호
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    • pp.1397-1403
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    • 2004
  • By recently study, GM (Gamipaemo-tang) treatment have worked well on the allergic asthma. The purpose of this study was effect of GM extract on helper T cell, major regulator of immune system. Splenic cells from 8-week BALB/c mice were cultured in GM containing media without activation for 48 hours. The MTS assay and flow cytometry study revealed that lymphocyte treated with GM were not effective on CD4+ T cells. Subsequently CD4+ T cells were isolated and cultured in GM containing media. Either GM were not effective on CD4+ T cell without APCs. By FACS scan analysis, the expression of INF-γ, IL-4 were down-regulated in the condition skewed Th1 and Th2 cells respectively, Using ELISA analysis, the expression of INF-γ is up-regulated and IL-4 is down-regulated in the condition skewed Th1, Th2 cells respectively. With RT-PCR analysis, the expression of mRNA for INF-γ is down-regulated and IL-4 is down-regulated in the condition skewed Th1 and Th2 cells respectively. The result suggests that GM inhibited the differetiation of Th2 cells significantly and indicates GM could enhance anti-allergic immune system.

당귀가 저산소로 손상된 성상세포의 gliosis 억제에 미치는 영향 (The Effect of the Water Extract of Angelica Sinens on Gliosis Repression of Astrocyte after Hypoxic injury)

  • 이승희;문성진;신진봉;허래경;성기문;양재훈;송봉근
    • 대한한의학회지
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    • 제29권1호
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    • pp.167-178
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    • 2008
  • Object : Gliosis becomes a physical and mechanical barrier to axonal regeneration. Reactive gliosis induced by hypoxic brain injury is involved with up-regulation of CD81 and GFAP. The current study was to examine the effect of the Angelica Sinens on CD81 and GFAP regulation after hypoxic brain injury in the astrocyte. Methods : MTT assay was performed to examine cell viability, and cell based ELISA, western blot and PCR were used to detect the expression of CD81 and GFAP. Results : The following results were obtained: 1. Using ELISA, western blot and PCR from the astrocyte after hypoxic injury, CD81 and GFAP expression was seen to have increased. 2. After the administration of Angelica Sinens extract to astrocyte following hypoxic injury, CD81 and GFAP expression was down regulated significantly. The water extract of Angelica Sinens prevented cell destruction by hypoxic induced with $CoCl_2$. Conclusion : These results indicate that Angelica Sinens could suppress reactive gliosis, which disturbs astrocyte regeneration after hypoxic brain injury by controlling the expression of CD81 and GFAP.

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포공영(蒲公英)이 MC/9 mast cell에서의 Th2 cytokine 발현에 미치는 영향 (Effects of TAM (Taraxacum mongolicum) on Th2 Cytokine Production in MC/9 Mast Cells)

  • 장문희;최재송;배나영;안택원
    • 사상체질의학회지
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    • 제24권1호
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    • pp.54-65
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    • 2012
  • 1. Objective : The purpose of this study is to investigate the effects of TAM (Taraxacum mongolicum) on Th2 cytokine production in MC/9 mast cells. 2. Methods : The effects of TAM was analyzed by ELISA and Real-time PCR in MC/9 mast cells. Levels of IL-5, IL-13 were measured using enzyme-linked immunosorbent assays(ELISA). mRNA levels of IL-4, IL-5, IL-6, IL-13 were analyzed with Real-time PCR. 3. Results : 1) TAM inhibited the IL-4 production significantly in comparison to PI-control group at concentration of $50{\mu}g/ml$, $100{\mu}g/ml$, $200{\mu}g/ml$. 2) TAM inhibited the IL-13 production significantly in comparison to PI-control group at concentration of $50{\mu}g/ml$, $100{\mu}g/ml$, $200{\mu}g/ml$. 3) TAM inhibited the IL-4 mRNA expression significantly in comparison to PI-control group at concentration of $100{\mu}g/ml$. 4) TAM inhibited the IL-5 mRNA expression significantly in comparison to PI-control group at concentration of $50{\mu}g/ml$, $100{\mu}g/ml$. 5) TAM inhibited the IL-6 mRNA expression significantly in comparison to PI-control group at concentration of $100{\mu}g/ml$. 6) TAM inhibited the IL-13 mRNA expression significantly in comparison to PI-control group at concentration of $100{\mu}g/ml$. 4. Conclusions : These results indicate that TAM (Taraxacum mongolicum) has the effect of decreasing the Th2 cytokine production in the MC/9 mast cell.

인삼(人蔘)이 중추신경계 손상 동물 모델의 재생에 미치는 영향 (The Effect of Ginseng Radix on Regeneration After Central Nervous System Injury)

  • 문형철;김윤욱;송봉근
    • Journal of Acupuncture Research
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    • 제24권6호
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    • pp.137-148
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    • 2007
  • Objectives : Following central nervous system (CNS) injury, inhibitory influences at the site of axonal damage occur. Glial cells become reactive and form a glial scar, know as gliosis. As well,myelin debris such as MAG inhibits axonal regeneration. Astrocyte-rich gliosis relates to up-regulation of GFAP and CD81, and eventually becomes a physical and mechanical barrier to axonal regeneration. It is postulated that when the astrocytic reaction is absent, regeneration of axons can occur. It was reported that treatment with anti CD81 antibodies enhanced functional recovery in rats with spinal cord injury. Methods : MAG is one of several endogenous axon regeneration inhibitors that limit recovery from central nervous system injury and disease. It was reported that molecules which block such inhibitors enhanced axon regeneration and functional recovery. Results : In this current study, the author investigated the effect of the water extract of Ginseng Radix on the regulation of CD81, GFAP and MAG which increases when gliosis occurs. MTT analysis was performed to examine cell viability, and cell based ELISA, Western Blot and PCR were used to detect the expression of CD81, GFAP and MAG. Immunohistochemistry was also performed to confirm in vivo. Conclusions : We observed that Ginseng Radix significantly down-regulates the expression of CD81, GFAP and MAG by means of cell based ELISA, Western Blot and PCR. In immunohistochemistry, expression of CD81, GFAP and MAG also decreased. Taken together, these results suggest that Ginseng Radix can be a candidate for regenerating CNS injury.

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Epidemiological Investigation of Asymptomatic Dogs with Leishmania Infection in Southwestern China Where Visceral Leishmaniasis is Intractable

  • Zhao, Gui-Hua;Yin, Kun;Zhong, Wei-Xia;Xiao, Ting;Wei, Qing-Kuan;Cui, Yong;Liu, Gong-Zhen;Xu, Chao;Wang, Hong-Fa
    • Parasites, Hosts and Diseases
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    • 제54권6호
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    • pp.797-801
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    • 2016
  • Heishui county, located in northwest Sichuan province, southwestern China, is an endemic area of zoonotic visceral leishmaniasis (VL) and is the most intractable area. VL is never destroyed in it. Asymptomatic dogs (Leishmania parasites have been diagnosed but clinically healthy) are considered to be a potential reservoir host in zoonotic VL area, and most can lead to infection of individuals, that is a new challenge for controlling VL in humans. The present study aimed to assess the Leishmania infection rate of asymptomatic dogs in Heishui county. Total 105 asymptomatic domestic dogs were gathered from 4 districts in Heishui county to investigate the infection rate with serological and molecular methods based on ELISA and kinetoplast minicircle DNA(kDNA) PCR, respectively. Out of 105 dogs, 44 (41.9%) were positive by more than 1 method; 21 (20.0%) were positive by ELISA, and 30 (28.6%) were positive by kDNA-PCR. Our study showed that Leishmania infection of domestic dogs which is clinically healthy is prevalent in the studied district, and the asymptomatic dogs infected by Leishmania may be the primary reason for the prevalence of visceral leishmaniasis in the area.

Saccharomyces cerevisae에서 한국산 겨우살이 유래 lectin A 및 B 유전자의 발현 (Expression of Recombinant Korean Mistletoe(KM) Lectin and B genes in Saccharomyces cerevisiae)

  • 최윤혁;김종배;양웅석;황철원
    • 생명과학회지
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    • 제14권5호
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    • pp.840-846
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    • 2004
  • 본 연구는 한국산 겨우살이 lectin유전자 (A 및 B chain) 을 효모 (Saccharmyces cerevisiae) 에 형질 전환시키는 시스템을 사용한 것으로, 효모내 효과적인 lectin유전자 발현을 위하여 유전자 상에 Kozak translation initiation sequence를 PCR을 이용 삽입, 변형시켜 재 클로닝 하였다. 변형된 lectin A 및 B 유전자를 포함하는 재조합 플라스미드는 S. cerevisiae INVSc (MATa, his3 $\Delta$l, leu2, trpl-289, ura3-52) 에 형질전환 되었다. 형질전환된 효모는 ABI 3700 system을 이용한 DNA 염기서열 분석을 통해 확인되었고 재조합 한국산겨우살이 lectin 발현을 위해 2% galactose를 사용하여 유도발현되었다. 재조합 lectin A 및 B 단백질은 SDS-PACE 및 western blotting 분석을 수행한 결과 약 29kDa 크기로 확인되었다. 재조합 lectin은 세포내 가용성 단백질 1mg중 1.24∼l.75 $\mu\textrm{g}$ 수준으로 발현되어짐을 ELISA 분석을 통해 확인하였다. 한편 lectin 유전자는 galartose 유도발현 후 36시간이 되 었을 때 발현량이 최대가 되었으며 lectin A 유전자의 경우 48 시간 이후에는 발현이 억제되었다.

우리나라 식물검역 격리재배 시스템과 2005-2012년 실적보고 (Plant quarantine isolated cultivation system in Korea and results of recorded in 2005-2012)

  • 이시원;박정안;이오미;신용길
    • 농업과학연구
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    • 제40권4호
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    • pp.281-287
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    • 2013
  • 우리나라의 격리재배는 102속, 약 250여종의 식물에 대해 실시하고 있으며, 직접경검법, 배양법, 선택배지, 생리생화학, ELISA 및 PCR 검사방법을 사용한다. 2005-2012년, 우리나라에서 수행된 격리재배는 총 8,307건이며 이중 구근류가 5,165건(62.2%)로 가장 많았고, 묘목류가2,119건(25.0%), 종자 796건(9.6%), 삽수 150건(1.8%), 접수 70건(0.8%) 및 기타 7건(0.1%) 이었다. 불합격 사례는 총 413건으로 약 4.97%였고, 발견된 병의 종류는 총 47종으로 나타났다. 종류별로는 바이러스가 27종으로 가장 많은 수를 차지했으며, 곰팡이 16종, 바이로이드 1종, Chromalveolata 1종 및 기타 2종으로 나타났다. 가장 많은 검역건을 올린 병원체는 Arabis mosaic virus (77건), Tobacco rattle virus (70건), Lily symptomless virus (46건), Penicillium expansum (46건)이다.