• 제목/요약/키워드: ELISA system

검색결과 326건 처리시간 0.025초

Exosome-mediated lnc-ABCA12-3 promotes proliferation and glycolysis but inhibits apoptosis by regulating the toll-like receptor 4/nuclear factor kappa-B signaling pathway in esophageal squamous cell carcinoma

  • Junliang Ma;Yijun Luo;Yingjie Liu;Cheng Chen;Anping Chen;Lubiao Liang;Wenxiang Wang;Yongxiang Song
    • The Korean Journal of Physiology and Pharmacology
    • /
    • 제27권1호
    • /
    • pp.61-73
    • /
    • 2023
  • Esophageal squamous cell carcinoma (ESCC) is a kind of malignant tumor with high incidence and mortality in the digestive system. The aim of this study is to explore the function of lnc-ABCA12-3 in the development of ESCC and its unique mechanisms. RT-PCR was applied to detect gene transcription levels in tissues or cell lines like TE-1, EC9706, and HEEC cells. Western blot was conducted to identify protein expression levels of mitochondrial apoptosis and toll-like receptor 4 (TLR4)/nuclear factor kappa-B (NF-κB) signaling pathway. CCK-8 and EdU assays were carried out to measure cell proliferation, and cell apoptosis was examined by flow cytometry. ELISA was used for checking the changes in glycolysis-related indicators. Lnc-ABCA12-3 was highly expressed in ESCC tissues and cells, which preferred it to be a candidate target. The TE-1 and EC9706 cells proliferation and glycolysis were obviously inhibited with the downregulation of lnc-ABCA12-3, while apoptosis was promoted. TLR4 activator could largely reverse the apoptosis acceleration and relieved the proliferation and glycolysis suppression caused by lnc-ABCA12-3 downregulation. Moreover, the effect of lnc-ABCA12-3 on ESCC cells was actualized by activating the TLR4/NF-κB signaling pathway under the mediation of exosome. Taken together, the lnc-ABCA12-3 could promote the proliferation and glycolysis of ESCC, while repressing its apoptosis probably by regulating the TLR4/NF-κB signaling pathway under the mediation of exosome.

사삼생맥산 추출물이 Balb/c mice의 면역활성에 미치는 효과 (Effect of Sasamsaengmaek-san Extract on Immunity Activity in Balb/c Mice)

  • 박채현;강재희;이현
    • 동의생리병리학회지
    • /
    • 제36권6호
    • /
    • pp.235-241
    • /
    • 2022
  • The purpose of this study was to investigate the immune activation effect of Sasamsaengmaek-san (SSSMS) consisted of a mixture of Adenophora triphylla var. japonica, Liriope platyphylla, Panax ginseng C. A. Meyer and Schisandra chinensis. in Balb/c mice. Measuring alanine aminotransferase (ALT) and aspartic acid transaminase (AST) levels in Balb/c mice was performed to analyze the cytotoxicity. Cytokines (IFN-γ, IL-2, IL-12) which regulate the immune activation in Balb/c mice were measured by enzyme-linked immunosorbent assay (ELISA). Activated T lymphocytes in peripheral blood mononuclear cell (PBMC), spleen, lymph nodes were analyzed by flow cytometry using percentages. All tests were compared with red ginseng 100 ㎍/mL (RG 100), which is the most used for immune activity. As a result, cytokine activity was significantly increased at SSSMS 300 group. Activated T lymphocytes in PBMC, spleen, lymph nodes were significantly increased at SSSMS 300 group. These results suggest that there is a possibility of SSSMS activating an immune system by activating the cytokines, and it is confirmed that SSSMS also effective for generation and differentiation of T, B lymphocytes which activate the immune response.

하수역학 구축을 위한 시료 전처리 기술과 신속검출기술 (Pretreatment and Rapid Detection Methods for Wastewater-Based Epidemiology)

  • 이재엽;이복진;제스민아터;안창혁;김일호
    • 한국물환경학회지
    • /
    • 제39권1호
    • /
    • pp.102-110
    • /
    • 2023
  • Wastewater Based Epidemiology (WBE) provides useful information not only on the use of illegal drugs in the community, but also on the presence of hygiene and health products and infectious pathogens in sewage facilities. As a consequence of the SARS-CoV-19 virus epidemic in 2019, monitoring the status of the infection is of utmost importance. SARS-CoV-19 was also detected in sewage, and the number and trend of infections in the community suggest that the application of the WBE system would be useful and appropriate. This study introduces a pre-treatment concentration method including viruses in sewage samples. A total of seven methods which were subdivided into methods for adsorption-extraction, ultra-filtration, PEG precipitation, and ultra-centrifugation, and the results for analyzing the recovery rates were included. Meanwhile, it is necessary to pay attention to rapid detection technologies which analyze infectious pathogens at the site of sewage facilities. These can include ELISA, FTIR, SERS, and biosensor based on the detection principle, and the characteristics, advantages, and disadvantages of each were summarized herein. If rapid detection technologies and accurate quantitative analyses are further developed, the use of sewage mechanics in response to pandemic viruses is expected to expand further.

보두산(寶豆散) 메탄올 추출물의 항염증 효과 (Anti-inflammatory Effect of Bodusan)

  • 김판준;윤현정;허숙경;김경애;김동완;김재은;박선동
    • 대한본초학회지
    • /
    • 제24권2호
    • /
    • pp.49-56
    • /
    • 2009
  • Objectives : Inflammation is important event in the development of vascular diseases including hypertension, atherosclerosis, and restenosis. Bodusan (BDS) was a traditional Korean herbal medicine and widely used in treatment of gastrointestinal complaint and stomach ulcer. The aim of this study was to determine whether BDS and its components inhibit production of nitrite, PGE2 and proinflammatory cytokines in lipopolysaccharide (LPS)-treated RAW 264.7 macrophages. Methods : Cytotoxic activity of BDS and its components on RAW 264.7 cells was using 5-(3caroboxymcrophages. eth-oxyphenyj)-2H-tetra-zolium inner salt (MTS) assay. The nitric oxide (NO) production was measured by Griess reagent system. And proinflammatory cytokines were measured by ELISA kit. The levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression were detected by western blot. Results : Our results indicated that BDS and its components significantly inhibited the LPS-induced NO and PGE2 production. Moreover. BDS and its components inhibited iNOS and COX-2 expression accompanied by an attenuation of TNF-${\alpha}$, IL-11${\beta}$, IL-6 and MCP-1 formation in macrophages. Conclusions: These results indicate that BDS and its components have potential as an anti-inflammatory agent.

Association of farmers' knowledge, attitude and practices with bovine brucellosis seroprevalence in Myanmar

  • Su Su Hlaing;Satoko Kubota;Kohei Makita;Ye Tun Win;Hnin Thidar Myint;Hiroichi Kono
    • Animal Bioscience
    • /
    • 제37권3호
    • /
    • pp.547-554
    • /
    • 2024
  • Objective: This study aimed to identify the relationship between bovine brucellosis prevalence, farmers' knowledge, attitude, practice (KAP), and social factors on migratory draft cattle and smallholder dairy farms in the central dry zone of Myanmar. Methods: This cross-sectional study was conducted on 54 migratory and 38 dairy cattle farms between August 2020 and February 2021. A structured questionnaire was used to identify farmers' behaviors. Bulk milk was sampled and tested using indirect enzyme-linked immunosorbent assay (I-ELISA). STATA 17 was used for all the analyses. Results: Migratory cattle farms had a higher farm level brucellosis prevalence (14.8%) than dairy farms (2.6%; χ2 = 3.75; df = 1; p = 0.05). Only 2.8% of the farmers had knowledge about brucellosis, while 39.1% and 41.6% had attitudes and farm practices with respect to brucellosis, respectively in the study area. Socio-economic attribute of training in animal husbandry (p<0.01), raising system (p<0.01), practice of separating the aborted cow (p<0.01) were negatively associated to brucellosis. The overall farm level brucellosis prevalence was strongly associated with cattle herd size (p = 0.01), free movement grazing practices (p<0.01), practice of self-removal of placental debris without using personal protective equipment (p<0.01) and farmers' attitudes towards eating cow placenta debris (p<0.01). Conclusion: Farmers had little knowledge of brucellosis. Attitudes and practices differed significantly between migratory and dairy farmers. Training and extension programs are necessary to make farmers aware of their KAP situation since livestock migration and the custom of eating cow placental debris contribute to the spread of brucellosis. Persistent efforts are required to reduce the adverse effects of brucellosis. Therefore, the study suggests that a feasible control intervention and public awareness campaigns need to be conducted regarding methods of preventing human exposure to brucellosis.

CagL 재조합 단백질 접종후에 Mongolian gerbil에서 나타나는 Helicobacter pylori 감염에 대한 반응 (Effect of Recombinant CagL Immunization on the Gastric Diseases Induced by Helicobacter pylori in Mongolian gerbils)

  • 박은정;장성일;최윤희;김진문;김애련;김지혜;우계형;유윤정;이성행;차정헌
    • 미생물학회지
    • /
    • 제48권2호
    • /
    • pp.109-115
    • /
    • 2012
  • Helicobacter pylori는 만성 위염, 소화성 궤양, 위암의 중요한 역학적 인자중 하나이다. H. pylori의 독성인자중 CagL은 숙주 세포와 H. pylori의 제 4형 분비기관(Type 4 secretion system)을 연결하는 adhesin으로 작용하는 섬모 단백질로 H. pylori가 발병하는데 중요한 역할을 하는 것으로 알려져 있다. 이번 연구는 저빌에 H. pylori를 감염시킨 동물 모델을 이용하여 CagL 재조합 단백질을 면역화시켰을 때 나타나는 효과를 평가하였다. 재조합 CagL은 클론되었고, 과발현시켜 정제하여 준비하였다 저빌은 H. pylori 감염 대조군과 H. pylori 감염 CagL 재조합 단백질 접종군으로 분류하였고, 접종시 알루미늄 애쥬번트를 사용하였다. 일주일 간격으로 4회 근육내 접종하였고, 마지막 접종 일주일 후, 모든 저빌에 H. pylori 7.13 균주를 $1{\times}10^9\;bacteria/500{\mu}l$ 농도로 위내 투여하였다. H. pylori 감염 6주째 모든 저빌을 희생하여 혈청 IgG 반응평가를 위한 ELISA를 실시하였고, 위에서는 집락화된 H. pylori의 수평가, 병리조직학적 평가 및 사이토카인 유전자발현을 조사하였다. CagL 재조합 단백질접종 일주일 후부터 H. pylori 감염 CagL 재조합 단백질 접종군의 혈청내 IgG 항체형성이 유의적으로 증가하였다. 위에서의 집락화된 세균수는 두군의 차이가 없었다. 저빌 체중에 대한 위무게 비율는 H. pylori 감염 CagL 재조합 단백질 접종군이 유의적으로 감소하였으나 병리조직학적 평가에서는 유의적인 차이는 확인하지 못하였다. 위에서의 IL-$1{\beta}$와 KC (IL-8 homologues)의 유전자발현 정도도 두 군사이에 유의적인 차이는 없었다. 이번 결과는 CagL 재조합 단백질의 접종은 IgG 항체형성은 효과적으로 자극하였지만 면역화된 숙주에서 세균 집락화의 감소 및 병변형성의 방어까지는 유도하지 못한 것으로 나타났으며, 앞으로 H. pylori 감염에 대해 유효한 면역 반응 및 질병 방어 효과를 나타내기 위해서 CagL을 포함한 다른 종류의 재조합 항원 사용 및 보조적으로 전신 면역 및 점막 면역을 효과적으로 유도하기 위해 안정성있는 애쥬번트의 사용을 고려해야 할 것으로 사료된다.

전기 검출 시스템을 이용한 Microfluidic Immuno-Sensor Chip (Microfluidic Immuno-Sensor Chip using Electrical Detection System)

  • 맹준호;이병철;조철호;고용준;안유민;조남규;이성환;황승용
    • KSBB Journal
    • /
    • 제21권5호
    • /
    • pp.325-330
    • /
    • 2006
  • Bio-MEMS를 기반으로 microfilter와 백금 전극이 내재되어 있는 microbiochip을 제작하였다. 우리는 이 chip으로 microbead에 indirect ELISA 방법으로 항원-항체를 반응시키고 전기 신호 검출 방법을 이용하여 반응 여부를 판단하였다. 이 때 신호 증폭을 위해 silver enhancer를 사용하였다. Chip 상에서 항원-항체 반응 조건을 최적화하기 위해 pH, temperature, flow time, flow rate, silver enhancer time을 결정하였다. 이렇게 최적화된 조건을 바탕으로 짧은 시간 안에 소량의 시료로 immunoassay를 성공적으로 수행할 수 있었다. 전기 신호 검출 방식을 사용함으로써 biosensor 장비의 소형화와 다중 시료 측정과 자동화를 biosensor에서 적용할 수 있는 가능성을 제시하였다.

생강감초탕(生薑甘草湯) 및 생강(生薑), 감초(甘草)가 생쥐 비장세포의 활성에 미치는 영향 (Effects of Senggang Gamcho-tang (SG), ZingberisRhizoma (ZR) and Glycyrrihizae Radix (GR) extracts in Senggang Gamcho-tang (SG) on Activity of Murine Splenocytes)

  • 이매;김진주;정희재;정승기
    • 대한한방내과학회지
    • /
    • 제32권3호
    • /
    • pp.323-333
    • /
    • 2011
  • Objectives : Zingberis Rhizoma (ZR) and Glycyrrihizae Radix (GR) have been widely used to prevent or treat allergic diseases. However, relatively little research has been conducted on the immune response to Senggang Gamcho-tang (SG) in a mixture of ZR and GR. The purpose of this study was to determine antiallergic effects of Senggang Gamcho-tang (SG) extracts ZR and GR on activity of murine splenocytes. Methods : This study was performed to investigate the effect of SG in mice, using in vitro experiments. Cells were treated with SG extract (1 ${\mu}g$/ml, 10 ${\mu}g$/ml and 100 ${\mu}g$/ml) plus Con A (2 ${\mu}g$/ml) and ZR (6.6 ${\mu}g$/ml and 66 ${\mu}g$/ml), GR (3.3 ${\mu}g$/ml and 33 ${\mu}g$/ml) respectively for 48 hrs. The production of IFN-${\gamma}$ and IL-4 were determined by ELISA. Results : No toxicity was found in splenocytes treated with SG extract for 48 hrs at the concentration of 0 ${\mu}g$/ml, 1 ${\mu}g$/ml, 10 ${\mu}g$/ml and 100 ${\mu}g$/ml, respectively. The production of IFN-${\gamma}$ increased to 23,968.7 pg/ml (p<0.001) while that of IL-4 significantly decreased to 14.1 pg/m l(p<0.05) only at the SG concentration of 100 ${\mu}g$/ml. With the GR concentration of 33 ${\mu}g$/ml in SG extract, the production of IFN-${\gamma}$ rose to 32,102.2 pg/ml (p<0.001) while that of IL-4 (p<0.05) declined to 34.2 pg/ml. However, there was no significant observation with regard to the effect of ZR on production of IFN-${\gamma}$ and IL-4. Conclusions : This study suggests an antiallergic effect of Senggang Gamcho-tang, which leads to the implication that SG extract may be used to enhance human immune function.

조절 T세포에 미치는 치자(梔子)의 효과 (Research on the Effect of Gardeniae Fructus on Regulatory T Cell Stimulation)

  • 서산;정희재;정승기
    • 대한한방내과학회지
    • /
    • 제31권2호
    • /
    • pp.189-200
    • /
    • 2010
  • Objectives : Regulatory T cells can reduce inflammation and allergic reactions through their inhibitory functions. Gardeniae Fructus(GF) is a Heat-clearing herb used in traditional Korean medicine, and a wide range of studies on its antiinflammatory effects are being carried out. The authors investigated the effect that Gardeniae Fructus has on regulatory T cells. Methods : The authors screened 14 herbs for their effects on regulatory T cells. 100mg of each herb were separately dissolved in 1ml of sterile saline and the supernatant was harvested after 10 minutes of centrifuge at 15,000 rpm. The supernatant was filtered through a 0.2 ${\mu}m$ syringe filter, and the resulting stock was refrigerated at $4^{\circ}C$. The stock was diluted before testing and used at a final concentration of $0.01{\mu}g/ml$. CD4+CD25+ T cells from healthy BALB/c spleens were used as natural regulatory T cells (nTreg), and CD4+CD25- T cells were used as reactive T cells. CD4+CD25+ and CD4+CD25- T cells were activated with anti-CD3e ($10{\mu}g/m{\ell}$)/anti-CD28 ($1{\mu}g/m{\ell}$) and cultured. IL-10 from supernatant of the culture medium was measured by IL-10 cytokine ELISA. The percentages, cell numbers, phenotype and function of CD4+CD25+ Treg cells were determined by flow cytometry. Results : Gardeniae Fructus was shown to be the most potent herb among the 14 herbs tested for suppressing CD4+CD25- reactive T cell proliferation by stimulating CD4+CD25+ natural regulatory T cells. Gardeniae Fructus induces IL-10 secretion increase by stimulating CD4+CD25+ natural regulatory T cells, and indirectly suppresses CD4+CD25- reactive T cell proliferation through increasing CD25 (IL-2 receptor $\alpha$) expression and thus promoting bonding with IL-2. Gardeniae Fructus did not directly affect CD4+CD25- reactive T cell proliferation. Conclusions : Gardeniae Fructus suppressed reactive T cell proliferation through inducing increases in IL-10 secretion and CD25 (IL-2 receptor $\alpha$) expression.

황정(黃精)으로부터 유래한 조다당류의 선천면역 활성에 의한 유방암 세포주 전이 억제 효과 (Antimetastatic Effects of Crude Polysaccharide Isolated from Polygonati Rhizoma on 4T1 Breast Cancer Cells by Activation of Innate Immune System)

  • 지해리;황덕상;이창훈;장준복;이진무
    • 대한한방부인과학회지
    • /
    • 제32권4호
    • /
    • pp.1-13
    • /
    • 2019
  • Objective: This study is aimed to investigate the anti-tumor metastasis by innate immunomodulating effects of crude polysaccharide isolated from Polygonati Rhizoma (CP-PR) on 4T1 breast cancer cells. Methods: CP-PR was isolated from Polygonati Rhizoma. Antimetastatic experiments were conducted in vivo mouse model by using 4T1 breast cancer cells. The cell viability of CP-PR was tested with normal spleen and 4T1 breast cancer cells. To observe the activation of macrophages with/without 4T1 breast cancer cells, production of tumor necrosis factor-${\alpha}$ ($TNF-{\alpha}$), interleukin-6 (IL-6), IL-10 and IL-12 were measured with enzyme-linked immunosorbent assay (ELISA), respectively. In addition, the lysis of YAC-1 cells and the production of granzymes were measured to observe the activation of natural killer (NK) cell. Results: Intravenous administration of CP-PR significantly inhibited metastasis of 4T1 breast cancer cells. In an in vitro cytotoxicity analysis, CP-PR affected the growth of normal spleen and 4T1 breast cancer cells above specific concentration. The production of $TNF-{\alpha}$, IL-6, IL-10 and IL-12 were significantly increased in macrophages with CP-PR. As compared with control, CP-PR showed significantly higher production of $TNF-{\alpha}$, IL-10 and IL-12 in macrophages co-cultured with 4T1 breast cancer cells. The lysis of YAC-1 cells and the production of granzymes were significantly up regulated by CP-PR. Conclusion: CP-PR appears to have considerable activity on the anti-metastasis by activation of innate immune system.