• Title/Summary/Keyword: ELISA system

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Analysis of Cytokine and Hormone on Sasang constitution (사상체질별 Cytokine 및 Hormone 분석)

  • Choi, Sun-Mi;Kim, Min-Hee;Chi, Sang-En;Kim, Hee-Soo;Sung, Hyun-Jea;Shin, Min-Kyu
    • Korean Journal of Oriental Medicine
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    • v.5 no.1
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    • pp.73-79
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    • 1999
  • In this research we proceeded experiments to find the basis which make it possible to explain the physical and pathological process of Sasang constitutional medicine, in the way substituting hematopoietic-immune system(essence of life, blood, Ki and mental faculties : 精血氣神) Under these suppositions, the essence of life(精) is the multipotent stem cell which has the possibility to be specialized to any cell, the Ki(氣), blood(血) and mental faculties(神) are inferred that they are formed from specialized the essence of life(精), the blood(血) is the red blood cells and etc. that appears as the result of the genesis of circulation system. The Ki(氣) is from specialized basic immunity, the mental faculties(神) means long-term memories or combined immunity. Cytokines can act as specilaizing, growing factors and particiate in extremly combined procedure being controlled by both positive and negative specializing signals. Blood gathering was carried out in the morning and on empty stomach. The plasma was seperated and Erythropoietin, Stem cell factor, Granulocyte-colony stimulaing factor, Tumor necrosis factor, interlukin-3, Interleukin-6 were measured with ELISA kit. According to the result of post analysis by Duncan, each constitution is different in SCF(stem cell factor), IL-6(interleukin-6), EPO(erythropoietin). The value of Stem cell factor is high in order of Soumin(少陰人), Soyangin(少陽人), and Taeumin(太陰人), The value of interleukin-6 is high in Taeumin, Soumin, and Soyangin. Erythropoietin is high in order of Soumin, Soyangin, and Taeumin.

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Effects of Kamijihwang-tang on Immune Cells and Cytokines in OVA-induced Asthmatic Mice (가미지황탕(加味地黃湯)이 천식모델 생쥐의 면역세포 및 사이토카인에 미치는 영향)

  • Rah, Do-Gyun;Park, Yang-Chun
    • The Journal of Korean Medicine
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    • v.29 no.2
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    • pp.7-20
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    • 2008
  • Objectives: The purpose of this study wasto examine the effects of Kamijihwang-tang (KJHT) extracts on immune cells and cytokines in ovalbumin (OVA)-induced asthmatic mice. Methods: C57BL/6 mice were injected, inhaled and sprayed with OVA for 12 weeks (four times a week) for asthma induction. Two experimental groups were treated with different concentrations of KJHT (400 mg/kg and 200 mg/kg) extracts and cyclosporine A (10 mg/kg) for the later 8 weeks. At the end of the experiment, the mice lung, PLN and spleen were removed and immune cells were analyzed by flow cytometer, IL-5, IL-13, eotaxin-2, $TNF-{\alpha}$ were analyzed by real-time PCR, serum histamine was analyzed by ELISA kit. Results: $CD3^+$, $CD3e^-/CCR3^+$, $CD3e^+/CD69^+$, $CD4^+/CD25^+$, $B220^+/IgE^+$, and $CD3e^+/DX5^+$ cells in lung, PLN, and spleen of the KJHT group (400 mg/kg) decreased compared with that of the control group. $CD3e^+/CD69^+$, $CD4^+/CD25^+$, and $CD3e^+/DX5^+$ cells in lung, PLN, and spleen of the KJHT group (200 mg/kg), CD3+, $CD3e^-/CCR3^+$ cells in lung and PLN of the KJHT group (200 mg/kg) and $B220^+/IgE^+$ cells in lung and spleen of the KJHT group (200 mg/kg) decreased compared with that of the control group. mRNA expression of IL-5, IL-13, eotaxin-2, and $TNF-{\alpha}$ in lung tissue of the KJHT groups (400 mg/kg and 200 mg/kg) decreased compared with that of the control group. Histamine in serum of the KJHT group (400 mg/kg) decreased compared with that of the control group. Conclusions: These results suggest that KJHT can be utilized effectively in treating asthma because it significantly reduces inflammatory cells and cytokines.

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Purification of Caudal-Related Homeodomain Transcription Factor and Its Binding Characterization

  • Jeong, Mi-Suk;Hwang, Eun-Young;Kim, Hyun-Tae;Yoo, Mi-Ae;Jang, Se-Bok
    • Journal of Microbiology and Biotechnology
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    • v.19 no.12
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    • pp.1557-1564
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    • 2009
  • Human CDX2 is known as a caudal-related homeodomain transcription factor that is expressed in the intestinal epithelium and is important in differentiation and maintenance of the intestinal epithelial cells. The caudal-related homeobox proteins bind DNA according to a helix-turn-helix structure, thereby increasing the structural stability of DNA. A cancer-tumor suppressor role for Cdx2 has been shown by a decrease in the level of the expression of Cdx2 in colorectal cancer, but the mechanism of transcriptional regulation has not been examined at the molecular level. We developed a large-scale system for expression of the recombinant, novel CDX2, in Escherichia coli. A highly purified and soluble CDX2 protein was obtained in E. coli strain BL21(DE3)RIL and a hexahistidine fusion system using Ni-NTA affinity column, anion exchange, and gel filtration chromatographies. The identity and secondary structure of the purified CDX2 protein were confirmed by MALDI-TOF MS, Western blot, and a circular dichroism analyses. In addition, we studied the DNA-binding activity of recombinant CDX2 by ELISA experiment and isolated human CDX2-binding proteins derived from rat cells by an immobilized GST-fusion method. Three CDX2-binding proteins were found in the gastric tissue, and those proteins were identified to the homeobox protein Hox-D8, LIM homeobox protein 6, and SMC1L1 protein.

Effects of Seonbangpaedok-tang and Sigyeongcheongpye-tang on PMA-induced Production of Airway Mucin and Expression of MUC5AC (선방패독탕(仙方敗毒湯)과 시경청폐탕(柴梗淸肺湯)이 NCI-H292 세포에서 PMA로 유발된 기도뮤신의 생성과 MUC5AC 발현에 미치는 영향)

  • Nam, Tae-Heung;Park, Yang-Chun
    • The Journal of Korean Medicine
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    • v.29 no.4
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    • pp.123-132
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    • 2008
  • Objective: In this study, the author tried to investigate whether revised Seonbangpaedok-tang (SPT) and Sigyeongcheongpye-tang (SCT) significantly effects both PMA-induced mucin production and MUC5AC gene expression from airway epithelial cells. Objective: In this study, the author tried to investigate whether revised Seonbangpaedok-tang (SPT) and Sigyeongcheongpye-tang (SCT) significantly affects both PMA-induced mucin production and MUC5AC gene expression from airway epithelial cells. Materials and Methods: Confluent NCI-H292 cells were pretreated for 30 min in the presence of SPT and SCT and treated with PMA (10 ng/$m{\ell}$), to assess the effects of the agents on PMA-induced mucin production by enzyme-linked immunosorbent assay (ELISA). Also, the effects of the agents on PMA-induced MUC5AC gene expression from the same cells were investigated. Possible cytotoxicities of the agent were assessed by measuring the rate of survival and proliferation of NCI-H292 cells after treatment of agents during 48 hrs. Results: (1) SPT and SCT did not show significant cytotoxicity to NCI-H292 cells; (2) SPT significantly inhibitedthe expression levels of PMA-induced MUC5AC gene in NCI-H292 cells. SCT slightly decreased the expression levels of PMA-induced MUC5AC gene; (3) SPT significantly decreased PMA-induced mucin production from NCI-H292 cells. However, SCT did not affect mucin production. Conclusion: Theseresults suggest that SPT can not only affect the production of mucin but also the expression of the mucin gene and this explains the traditional use of SPT in oriental medicine. The effects of SPT and SCT with their components should be further investigated using animal experimental models that reflect pathophysiology of airway diseases via ongoing studies.

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Alteration of Plasma ${\beta}$-Nerve Growth Factor Concentration in Depressed Patients with Suicidal Attempt (자살을 시도한 우울증 환자에서 혈장 ${\beta}$-Nerve Growth Factor 농도의 변화)

  • Shim, Se-Hoon;Won, Seong-Doo;Lee, Bun-Hee;Han, Chang Su;Yang, Jong-Chul;Kwon, Young-Joon;Kim, Yong-Ku
    • Korean Journal of Biological Psychiatry
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    • v.13 no.2
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    • pp.95-102
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    • 2006
  • Object : Nerve growth factor(NGF) is a protein involved in neuronal survival and plasticity in the central nervous system, which might play an important role in stress, depression and suicide. This study was performed to determine whether there is an alteration in plasma NGF concentrations in depressed patients with suicidal attempt. Methods : The subjects were 32 depressed patients who attempted suicide and admitted in emergency room. Forty-four hospitalized non-suicidal depressive patients and the 30 normal controls were closely matched with the suicidal group in terms of age and sex. Individuals in all 3 groups were evaluated independently by a semi-structured interview for the purpose of establishing a DSM-IV criteria diagnosis. The severity of depressive symptoms was evaluated using Hamilton depression rating scale(HDRS). The severity of the suicidal behavior was evaluated by Weisman and Worden's risk-rescue rating(RRR) system and the Lethality Suicide Attempt Rating Scale(LSARS). Plasma NGF level was measured by the enzyme linked immunosorbent assay(ELISA) method. Results : There were no statistically significant differences of the plasma NGF levels among groups. LSARS and RRR did not reveal any significant correlation with ${\beta}$-NGF level in suicidal depressive patients. Conclusion : This study do not support an association between ${\beta}$-NGF and suicidal depression. However it is necessary to investigate this association through other route such as postmortem brain.

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Effects of Seonpye-tang on OVA-Induced Asthmatic Mice (천식모델생쥐에서 선폐탕(宣肺湯)의 면역조절 효과)

  • Lee, Ju-Guan;Park, Yang-Chun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.23 no.2
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    • pp.457-463
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    • 2009
  • This study aimed to evaluate the anti-asthmatic effects of Seonpye-tang (SPT) using OVA-induced asthmatic mice model. Scavenging activity of SPT on DPPH free radical and SOD-like activity of SPT were measured at final concentration 62.5, 125, 250, 500 (${\mu}g/m{\ell}$), RBL-2H3 cells were treated with DNP IgE for 24hr, and treated with SPT (1, 10, 100 ${\mu}g/m{\ell}$) for 1hr, followed by treatment with DNP-HSA for 1hr at $37^{\circ}C$. The level of IL-4 and TNF-${\alpha}$ were measured by ELISA. Asthmatic mice model was conducted by repeated challenge of OVA using C57BL/6 mice. Each group was treated with distilled water, SPT (400 mg/kg and 200 mg/kg) extract or cyclosporin A (10 mg/kg) for the later 8 weeks. Immune cells subpopulation, eotaxin, IL-5 and TNF-${\alpha}$ in BALF were analyzed. SPT dose-dependently increased Scavenging activity on DPPH free radical and SOD-like activity. SPT significantly ameliorated the increase of total cells number and eosinophil including of immune cell subpopulation of $CD3^+/CD69^+$, $CCR3^+$, $B220^+/CD22^+$, $B220^+/CD45^+$ and $B220^+/IgE^+$ cells in BALF comparing to control group. Eotaxin and IL-5 level in BALF were significantly decreased by SPT. These results strongly suggest that SPT would be a effective candidate for herbal-originated anti-asthmatic drug. However, this drug should be further studied for characterization of the accurate action and underlying mechanisms using variant disease model in the future.

A Rapid and Sensitive Screening System for Human Type I Collagen with the Aim of Discovering Potent Anti-Aging or Anti-Fibrotic Compounds

  • Hashem, Md. Abul;Jun, Kyu-Yeon;Lee, Eunyoung;Lim, Soyun;Park Choo, Hea-Young;Kwon, Youngjoo
    • Molecules and Cells
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    • v.26 no.6
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    • pp.625-630
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    • 2008
  • This study was undertaken with the aim of developing an easy and quick means of analyzing the effect of various compounds on the synthesis and secretion of human type I collagen at the protein level. A modification of the ELISA method was used on HFF-1 cells. For the proof of concept, we used thirteen compounds most of which are known to be antioxidants. Each compound was tested at concentrations of 0, 10 and $100{\mu}m$ on HFF-1 cells for 24 h. Thirteen sets of experiments for each compound were performed in ANOVA with three replicates. Duncan multiple range test (DMRT) was used to compare the mean values obtained from the treatment groups. From the results it was concluded that Vitamin C, undecylenic acid, conjugated linoleic acid, glycolic acid, and citric acid at $100{\mu}m$ concentration could be used for anti-wrinkling or protection from premature aging, which requires enhancement of collagen synthesis. Lactic acid, EGCG, resveratrol, and retinol that can inhibit collagen synthesis effectively in a dose-dependent manner may be used for anti-fibrosis treatment purposes.

Molecular and Circulatory Expression of Insulin Growth Factors in Indian Females with Advanced Cervical Cancer

  • Sharma, Manoj;Satyam, Abhigyan;Abhishek, Ashu;Khan, Rehan;Rajappa, Medha;Sharma, Alpana
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.12
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    • pp.6475-6479
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    • 2012
  • Background: Recent studies have demonstrated an association between insulin growth factor (IGF) and insulin growth factor binding protein-3 (IGFBP-III) serum levels and increased risk for various cancers. However, little information is available on clinical implications of the IGF system in Indian patients with cervical cancer. This study explored associations by analyzing their expression profiles in cervical cancer cases. Materials and Methods: Totals of 50 patients with advanced cervical cancer and 40 healthy controls were enrolled. Human papillomavirus (HPV) and cervical biopsy sample were obtained from all participating women. Circulatory levels were estimated by ELISA and the tissue expression was assessed using RT-PCR and Western blotting. Results: Levels of IGF-I and II showed significant increase whereas IGFBP-III showed significant decline in all patients as compared to controls. Spearman correlation analysis between IGFs and HPV status showed significant correlations. Conclusions: We demonstrated elevated circulating levels and tissue expression of IGF-I and IGF-II in advancer cancer cervix patients, as compared with controls, with a converse trend being apparent for IGFBP-III. In future, associations of the IGF system and clinical outcome of cervical cancer patients in post treatment samples might point to significance in disease mapping as a prognostic marker after validation with a larger patient series.

The Effect of the Radix Ginseng on Expression of CDSI and GFAP Following Hypoxic Injury on Central Nervous System (인삼(人蔘)이 허혈성 중추신경 손상으로 발현 증가된 CD81 및 GFAP에 미치는 영향)

  • Seo, Jong-Hoon;Song, Bong-Gun;Lyu, Yeoung-Su;Kang, Hyung-Won;Kim, Tae-Heon
    • Journal of Oriental Neuropsychiatry
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    • v.20 no.1
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    • pp.177-197
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    • 2009
  • Objectives : Gliosis disturbs recovery of damaged astrocytes following central nervous system(CNS) injury. Gliosis relates to up-regulation of CD81 and GFAP. In glial cells at injured CNS, the expression of CD81 and GFAP is increased. It is possible that when the gliosis formation is suppressed, regeneration of oxons can occur. According to the recent study, the treatment with anti CD81 antibodies enhanced functional recovery in rats with spinal injury. So, the author studies the effect of water extract of Radix Ginseng on regulation of CD81 and GFAP with CNS injury. Methods : In the cell study, hypoxic damage was induced by CoC12. And according to Longa et al, cerebral ischemia was made by middle cerebral artery occlusion in the rat. Cross sections of rat brain were examined under microscope. MTT analysis was performed to examine cell viability, cell based ELISA, Western Blot and PCR were used to detect the expression of CD81 and GFAP. Results : The following results were obtained. 1. We found that CD81 and GFAP were decreased in hypoxic injured cells following Radix Ginseng administration. 2. We injected the extract of Radix Ginseng to the middle cerebral artery occlusion in rats, and the immunohistochemistry analysis showed that CDS1 and GFAP were decreased. Conclusions : These results show that the extract of Radix Ginseng could suppress the gliosis formation and prevent cell death, by controlling the expression of CDS1 and GFAP. Therefore, Radix Ginseng could be a useful to regenerate CNS injury.

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Lysate of Probiotic Lactobacillus plantarum K8 Modulate the Mucosal Inflammatory System in Dextran Sulfate Sodium-induced Colitic Rats

  • Ahn, Young-Sook;Park, Min Young;Shin, Jae-Ho;Kim, Ji Yeon;Kwon, Oran
    • Food Science of Animal Resources
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    • v.34 no.6
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    • pp.829-835
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    • 2014
  • Inflammatory bowel disease (IBD) is caused by dysregulation of colon mucosal immunity and mucosal epithelial barrier function. Recent studies have reported that lipoteichoic acid (LTA) from Lactobacillus plantarum K8 reduces excessive production of pro-inflammatory cytokine. In this study, we investigated the preventive effects of lysate of Lb. plantarum K8 in dextran sulfate sodium (DSS)-induced colitis. Male Sprague-Dawley rats were orally pretreated with lysate of Lb. plantarum K8 (low dose or high dose) or live Lb. plantarum K8 prior to the induction of colitis using 4% DSS. Disease progression was monitored by assessment of disease activity index (DAI). Histological changes of colonic tissues were evaluated by hematoxylin and eosin (HE) staining. Tumor necrosis factor-alpha (TNF-${\alpha}$), interleukin-6 (IL-6) levels were measured using enzyme-linked immunosorbent assay (ELISA). The colon mRNA expressions of TNF-${\alpha}$, IL-6, and toll like receptor-2 (TLR-2) were examined by quantitative real-time-transcription polymerase chain reaction (qPCR). Lysate of Lb. plantarum K8 suppressed colon shortening, edema, mucosal damage, and the loss of DSS-induced crypts. The groups that received lysate of Lb. plantarum K8 exhibited significantly decreased levels of the pro-inflammatory cytokines TNF-${\alpha}$ and IL-6 in the colon. Interestingly, colonic expression of toll like receptor-2 mRNA in the high-dose lysate of Lb. plantarum K8 group increased significantly. Our study demonstrates the protective effects of oral lysate of Lb. plantarum K8 administration on DSS-induced colitis via the modulation of pro-inflammatory mediators of the mucosal immune system.