• Title/Summary/Keyword: ELISA method

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Diagnostic Value of Fecal Calprotectin as a Screening Biomarker for Gastrointestinal Malignancies

  • Khoshbaten, Manouchehr;Pishahang, Parinaz;Nouri, Mohammad;Lashkari, Alireza;Alizadeh, Mahasti;Rostami-Nejad, Mohammad
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.4
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    • pp.1667-1670
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    • 2014
  • Background: Calprotectin in feces seems to be a more sensitive marker for gastrointestinal (GI) cancers than fecal occult blood, but its specificity may be too low for screening average risk populations. This study aims at evaluating the diagnostic value of fecal calprotectin as a screening biomarker for GI malignancies. Materials and Methods: In a case-control study, 100 patients with GI malignancies (50 patients with colorectal cancer and 50 patients with gastric cancer) and 50 controls were recruited in Tabriz Imam Reza and Sina hospitals during a 24-month period. One to two weeks after the last endoscopy/colonoscopy, fecal specimens were collected by the patients and examined by ELISA method for quantitative measurement of calprotectin content. The results were compared between the three groups. Results: The mean fecal calprotectin level was $109.1{\pm}105.3$ (2.3-454.3, median:74), $241.1{\pm}205.2$ (3.4-610.0, median:19.3) and $45.9{\pm}55.1{\mu}g/g$ (1.3-257.1, median:19.3) in gastric cancer, colorectal cancer and control group, respectively, the differences being significant (p<0.001) and remaining after adjustment for age. The optimal cut-off point for fecal calprotectin was ${\geq}75.8{\mu}g/g$ for distinguishing colorectal cancer from normal cases (sensitivity and specificity of 80% and 84%, respectively). This value was ${\geq}41.9{\mu}g/g$ for distinguishing gastric cancer from normal cases (sensitivity and specificity of 62%). Conclusions: Our results revealed that fecal calprotectin might be a useful and non-invasive biomarker for distinguishing colorectal cancer from non-malignant GI conditions. However, due to low sensitivity and specificity, this biomarker may not help physicians distinguishing gastric cancer cases from healthy subjects.

Clinical Value of Eukaryotic Elongation Factor 2 (eEF2) in Non-small Cell Lung Cancer Patients

  • Sun, Hong-Gang;Dong, Xue-Jun;Lu, Tao;Yang, Ming-Feng;Wang, Xing-Mu
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.11
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    • pp.6533-6535
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    • 2013
  • Background: The purpose of this study was to evaluate a new type of tumor biomarker, eukaryotic elongation factor 2 (eEF2), in serum for the early diagnosis, confirmative diagnosis as well as assessment of treatment of non-small cell lung cancer (NSCLC). Methods: 130 patients with NSCLC and 50 healthy individuals undergoing physical examination in our hospital provided the observation and healthy control groups. An enzyme linked immune sorbent assay (ELISA) method was applied to determine serum eEF2 levels. Serum neuron specific enolase (NSE) and squamous cell carcinoma antigen (SCC) levels in the observation group were assessed with an automatic biochemical analyzer. Results: The median levels of eEF2 in the serum of NSCLC patients was found to be significantly higher than the healthy control group (p < 0.01) and it was markedly higher in stages III, IV than stages I, II (p < 0.05). eEF2 was higher with tumor size ${\geq}2$ cm than <2 cm (P< 0.01). Furthermore, two weeks after surgery patients showed a significant trend for eEF2 decrease (p < 0.05). Conclusions: The eukaryotic elongation factor 2 (eEF2) has certain clinical values for early diagnosis, verification, and prognosis as well as classification of lung cancer patients.

The Effects of Yeouigeumhwang-san on Anti-Inflammation and Anti- Propionibacterium acnes (여의금황산(如意金黃散)이 여드름 유발균과 염증에 미치는 영향)

  • Yoo, Jin-Gon;Seo, Hyeong-Sik
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.20 no.2 s.33
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    • pp.77-88
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    • 2007
  • Objectives : This experimental study was performed to investigate the effects of Yeouigeumhwang-san(YUGHS) on anti-inflammation and anti-Propionibacterium acnes. Methods : The cytotoxicity of YUGHS about viability of Raw 264.7 cell was tested by using a colorimetric tetrazolium assay(MTT assay). To investigate the anti-inflammatory effets of YUGHS on LPS-induced macrophage Raw 264.7 cell, we used ELISA kit and Western blots. Inhibitory effects of YUGHS on Propionibactrium acnes were investigated by using paper disk diffusion method. Results : 1. YUGHS has no cytotoxicity under 50 ${\mu}g/ml$ concentration but over 50 ${\mu}g/ml$ has a little cytotoxicity in Raw 264.7 cell. 2. Concentration of 100 ${\mu}g/ml$ YUGHS inhibited the production of NO in the Raw 264.7 cell stimulated with LPS. 3. All concentrations of YUGHS did not inhibit the production of $TNF-{\alpha}$ in the Raw 264.7 cell stimulated with LPS. 4. All concentrations of YUGHS significantly inhibited the production of $PGE_2$ in the Raw 264.7 cell stimulated with LPS. 5. YUGHS did not inhibit the expression of COX-2 but concentration of 50 ${\mu}g/ml$ YUGHS inhibited iNOS expression in the Raw 264.7 cell stimulated with LPS. 6. YUGHS has the effect of blocking $NF-{\kappa}B$ into nucleus in LPS-induced macrophage Raw 264.7 cell 7. YUGHS did not have the inhibitory effect of Propionibactrium acnes. Conclusions : These results indicate that Yeouigeumhwang-san has anti-inflammatory effets. If further study is performed, the use of Yeouigeumhwang-san will be valuable and benificial in the therapy of acnes.

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Experimental studies on the Whitening and Anti-allergic effect of various Herbal medicines that clear heat (수종의 청열약물의 미백 및 항알러지 효과에 대한 실험적 연구)

  • Lee, Myoung-Gyu;Kim, Kyung-Jun
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.20 no.3
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    • pp.71-81
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    • 2007
  • Objective : In this study, herbal medicine(GJE, Gardenia jasminoides Ellis; HCT, Houttuynia cordata Thunb.; CIL, Chrysanthemum indicum Linne; PMS,Paeonia moutan Sims, P. subfruticosa Makino; APL, Agrimonia pilosa Ledebour) were screened for their inhibitory activities against Tyrosinase and PMA plus A23187-induced $TNF-{\alpha}$, IL-6, IL-8 productions in HMC-1 cells to reveal their skin -whitening and anti-allergic effect. Method : To investigate Tyrosinase inhibition we treated Mushroom Tyrosinase(Fluka, 93898) $10{\mu}{\ell}$ and 7.5mM Tyrosine (Sigma, T3754) $20{\mu}{\ell}$ with 80% ethanol medicine extracts. Then we observed 96well micro plate extinction at 490nm. In the next experiment, to investigate Anti-allergic effect we blended cultured Human Mast Cells(HMC-1) with medicine extracts. We treated the blended solution with Phorbol 12-myristate 13-acetate(PMA) and A23187, then observed $TNF-{\alpha}$, IL-6, IL-8 by ELISA (enzyme-linked immunosorbent assay) at 450nm. Results : In inhibiting Tyrosinase the results are as follows. 1. We observed 22% inhibition of Mushroom Tyrosinase at $500{\mu}g/m{\ell}$ concentration of GJE extracts. 2. We also could observe that the decreased Mushroom Tyrosinase activities in HCT, CIL extracts. In inhibiing $TNF-{\alpha}$, IL-6, IL-8 productions in HMC-1 cells the results are as follows. 1. Of the extracts examined, HCT, PMS, APL extracts showed over 50% inhibitions of Cytokines at $200{\mu}g/m{\ell}$ concentration. 2. In particular, APL extracts showed the best inhibitory effect on Cytokine productions in a dose-dependent manner. Conclusion : These results suggest that GJE extracts contributes to the anti melanin activities and represent a potential source of whitening agent. Thus these herbal medicines suggest novel drugs on anti-allergic effects.

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Effects of Gyeonbi-Tang Treatment on the Monosodium Iodoacetate-induced Mild Osteoarthritis in Rats (견비탕(蠲痺湯)이 Monosodium Iodoacetate로 유발된 골관절염의 초기변화에 미치는 영향)

  • Park, Dong-Soo;Kim, Soon-Joong;Jeong, Su-Hyeon;Seo, Il-Bok
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.25 no.1
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    • pp.84-91
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    • 2011
  • This study was to investigate the effects of Gyeonbi-Tang Treatment on the monosodium iodoacetate(MIA)-induced mild osteoarthritis in rats. Arthritis was induced by injection of MIA(0.25 mg) into knee joints of rats. Arthritic rats were divided into control(n=8) and treated(n=8) group. Control group was taken distilled water and treated group was taken extracts of Gyeonbi-Tang by orally for 20days. Body weights were measured at 0, 5, 10, 15, 20 days after MIA injection. At the end of experiment(20day after MIA injection), gross and histopathological examination on the articular structures of knee joints were performed. Proteoglycan(PG) contents in articular cartilages were analysed. And also, tumor necrosis factor-${\alpha}$(TNF-${\alpha}$) and interleukin-1${\beta}$(IL-1${\beta}$) contents in synovial fluids were measured by ELISA method. Body weights of the treated group were significantly increased compared with control group at 15, 20 days after injection. Grossly, the severity of osteoarthritis in the treated group were alleviated compared with control group. PG contents in articular cartilages of the treated group were increased compared with control group. Histopathologically, osteoarthritic scores of the treated group was decreased compared with the control group. TNF-${\alpha}$ contents in synovial fluids of the treated group were significantly decreased compared with control group. On the basis of these results, we concluded that Gyeonbi-Tang Treatment has anti-arthritic effects on the monosodium iodoacetate-induced mild osteoarthritis in rats. And it's effects were related with reduced secretion of TNF-${\alpha}$ from synovial membranes.

Detection of Peripheral Blood Telomerase Activity from Gastric Cancer Patients (위암 환자의 혈액에서 Telomerase 활성도 검출의 의의)

  • Park Ki Ho;Jung Soon Jai;Yu Young Woon;Park Sung Hwan;Lee Han Il;Joo Dae Hyun;Park Ki Hyuk;Choi Dong Rak;Jeon Chang Ho
    • Journal of Gastric Cancer
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    • v.3 no.4
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    • pp.201-205
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    • 2003
  • Purpose: Telomerase activity is generally absent in primary cell cultures and normal tissues. Telomerase is known to be induced upon immortalization or malignant transformation of human cells. Telomerase activity can be increased in immature lymphocytes and activated lymphocytes, but it is not detected in the peripheral blood of normal persons. The authors analyzed peripheral blood telomerase from patients of gastric cancer to evaluate the possibility of using it for diagnosis and as a prognostic factor. Materials and Methods: We obtained blood samples from 11 inflammatory patients and 64 gastric cancer patients. The telomerase activity was measured using the [PCR-ELISA] method. The results were correlated with the T, N, M stage, cell differentiation, vascular, neural, and lymphatic invasion, tumor size, and tumor location. Results: In the 11 inflammatory patients, telomerase activity was not detected while in the gastric cancer patients, a positive rate of $28.1\%$ was noted. The peripheral telomerase activity was not related with tumor size, tumor site, lymphatic and vascular invasion, stage, or histologic differentiation. Conclusion: The peripheral blood telomerase activity for patients of gastric cancer can be utilized as a marker for the diagnosis of not only advanced gastric cancer, but also relatively early stage gastric cancer, but not as a prognostic factor.

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p53 Gene Mutation, Tumor p53 Protein Overexpression, and Serum Anti-p53 Antibody in Patients with Gastric Cancer (위암 환자에 있어서 p53 유전자 돌연변이, 종양 p53 단백질 과발현 및 혈청 p53 항체)

  • Bong Jin-gu;Lee Myung-Hoon;Song Kyung-Eun;Kim Taebong;Yu Wansik
    • Journal of Gastric Cancer
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    • v.3 no.4
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    • pp.206-213
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    • 2003
  • Purpose: The clinical implication of p53 mutation in gastric cancer is still unclear, as shown by the discordant results that continue to be reported in the literature. Materials and Methods: To assess p53 gene mutation, tumor p53 overexpression, and serum anti-p53 antibody, we employed a polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) analysis, an immunohistochemistry using monoclonal antibody DO-7, and an enzymelinked immunosorbent assay (ELISA), respectively. Results: Of 169 surgical specimens of gastric cancer, mutation at exon $5\∼8$ of the p53 was identified in 33 ($19.5\%$) and was significantly correlated with lymph node metastasis. Overexpression of p53 was found in 62 specimens ($36.7\%$) and had a significant correlation with tumor differentiation. Serum anti-p53 antibody was positive in 18 patients ($10.7\%$). Twenty-three of the mutated tumors ($69.7\%$) and 39 of the non-mutated tumors ($28.7\%$) displayed immunoreactivity. Twelve of the immunopositive tumors ($19.4\%$) and 6 of the immunonegative tumors produced anti-p53 antibody. These differences were statistically significant (P<0.001 and P=0.005, respectively). There was no significant difference in survival according to the mutation of p53. Conclusion: Mutation and overexpression of p53 can be easily detected by immunohistochemistry. However, standardization of the immunohistochemical staining method, as well as guidelines for interpreting the stained result, will produce concordant results and thereby improve clinical application.

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Simplified elastic design checks for torsionally balanced and unbalanced low-medium rise buildings in lower seismicity regions

  • Lam, Nelson T.K.;Wilson, John L.;Lumantarna, Elisa
    • Earthquakes and Structures
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    • v.11 no.5
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    • pp.741-777
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    • 2016
  • A simplified approach of assessing torsionally balanced (TB) and torsionally unbalanced (TU) low-medium rise buildings of up to 30 m in height is presented in this paper for regions of low-to-moderate seismicity. The Generalised Force Method of Analysis for TB buildings which is illustrated in the early part of the paper involves calculation of the deflection profile of the building in a 2D analysis in order that a capacity diagram can be constructed to intercept with the acceleration-displacement response spectrum diagram representing seismic actions. This approach of calculation on the planar model of a building which involves applying lateral forces to the building (waiving away the need of a dynamic analysis and yet obtaining similar results) has been adapted for determining the deflection behaviour of a TU building in the later part of the paper. Another key original contribution to knowledge is taking into account the strong dependence of the torsional response behaviour of the building on the periodic properties of the applied excitations in relation to the natural periods of vibration of the building. Many of the trends presented are not reflected in provisions of major codes of practices for the seismic design of buildings. The deflection behaviour of the building in response to displacement controlled (DC) excitations is in stark contrast to behaviour in acceleration controlled (AC), or velocity controlled (VC), conditions, and is much easier to generalise. Although DC conditions are rare with buildings not exceeding 30 m in height displacement estimates based on such conditions can be taken as upper bound estimates in order that a conservative prediction of the displacement profile at the edge of a TU building can be obtained conveniently by the use of a constant amplification factor to scale results from planar analysis.

Effect of Injin Fraction on Hepatic Fibrosis induced by $TGF-{\beta}1$ (인진이 $TGF-{\beta}1$ 유도성 간섬유화에 미치는 영향)

  • 신성만;김영철;이장훈;우흥정
    • The Journal of Korean Medicine
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    • v.22 no.3
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    • pp.141-155
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    • 2001
  • Objective : The aim of this study is to investigate the effect of Injin fractions on hepatic fibrosis induced by $TGF-{\beta}1$. Method : $TGF-{\beta}1$ mRNA, protein, $TGF-{\beta}1$ receptor, Smad family and PAI-I mRNA were studied in HepG2 cell, and the proliferation, connective tissue growth factor, fibronectin and collagen type I mRNA in T3891 fibroblast by quantitative RT-PCR, ELISA and thymidine incorporation assay. Results : On $TGF-{\beta}1$ mRNA and protein synthesis in HepG2, $H_2O$, butanol and hexane fractions of Injin showed inhibitory effect in a dose-dependent way. In the study on $TGF-{\beta}1$ receptor, Smad family and PAI-1 mRNA in HepG2, $H_2O$, butanol and hexane fraction of Injin showed inhibitory effect on the expression of PAI-1 in a dose-dependent way. On the proliferation of T3891 fibroblast induced by $TGF-{\beta}1$, $H_2O$, ethylacetate and butanol fractions of Injin showed inhibitory effect. In the study on the factors affected by $TGF-{\beta}1$, $H_2O$, ethylacetate and butanol fractions of Injin showed inhibitory effect on CTGF, and $H_2O$, butanol, chloroform and hexane fractions showed inhibitory effect on the expression of collagen type I, whereas no fraction showed inhibitory effect on the expression of fibronectin Conclusion : These results show that each fraction of Injin acts as a fibrosis inhibitory factor by itself or in combination, ultimately inhibiting liver cirrhosis.

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Reduction of Allergic Potential of Meju by Three Step Fermentation (3단계 발효에 의한 콩 알레르기성의 저하)

  • Ryu, Chung-Ho;Lee, Jeong-Ok;Son, Dae-Yeul
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.8
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    • pp.1066-1071
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    • 2012
  • In this study we investigated the change in antigenicity and allergenicity of Meju, a traditional Korean soybean product, by fermentation via 3 different microorganisms. The steamed soybeans were fermented with Lactococcus lactis subsp. lactis and/or Aspergillus oryzae and/or Bacillus subtilis. Proteins in soybean were degraded after fermentation. Antigenicity or allergenicity were analysed by immunoblotting and ELISA using soybean protein-specific polyclonal antibodies or soybean allergic patient sera. The best degradation was achieved by three step fermentation using nisin-producing Lactococcus lactis subsp. lactis IFO12007, A. oryzae and B. subtilis. Allergenicity and antigenicity were also starkly reduced after three step fermentation. The three-step fermentation method developed in our lab suggests an excellent alternative to reduce the allergenicity of soybeans.