• Title/Summary/Keyword: EIA-kit

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Inhibitory Effect of Injinchunggan-tang(Yinchenqinggan-tang) on Fibrogenesis of Rat Hepatic Stellate Cells (인진청간탕(茵蔯淸肝湯)이 간성상세포의 섬유화 억제에 미치는 영향)

  • Lee, Hong-Ri;Kim, Young-Chul;Woo, Hong-Jung;Lee, Jang-Hoon
    • The Journal of Internal Korean Medicine
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    • v.30 no.1
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    • pp.74-84
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    • 2009
  • Objectives : This study was performed to investigate the anti-fibrogenic effect of Injinchunggan-tang on cultured rat hepatic stellate cells. Materials and Methods : Hepatic stellate cells(HSC-T6) were treated with various concentrations of Injinchunggan-tang extract for 24, 48, and 72 hours. The extraction was done with distilled water. After the treatment, cell viability, proliferation, procollagen levels and the mRNA of the TIMP-1, TIMP-2, and ASMA were measured by using MTT assay. BrdU assay, procollagen type I C-peptide EIA kit and RT-PCR. Results : The proliferation, mRNA expression and synthesis of collagen of the hepatic stellate cells were inhibited by Injinchunggan-tang treatment in a dose-dependent manner. This indicates the prescription has inhibitory effect on fibrogenesis of the liver by regulating the fibrogenesis associated genes in transcription. Cell viability was inhibited in time- and dose-dependent manners. It seemed that the drug should be used with sufficient dose to acquire treatment effect. Conclusion : These results suggest that Injinchunggan-tang is beneficial in the treatment of cirrhotic patients as well as for the patients with chronic hepatitis.

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Effect of Sleep-restriction on Salivary Adrenal Hormones in Korean Adolescent (수면제한이 청소년의 타액 호르몬에 미치는 영향)

  • Cho, Yoon-Seong;Je, Jun-Tae;Lee, Sang-Kwan
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.25 no.2
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    • pp.311-317
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    • 2011
  • The goal of this study was to clarify effects of sleep restriction on a diurnal rhythm of salivary cortisol and DHEA levels in Korean adolescents.83 middle school students were recruited to participate in restricted sleep period group (less than 4h/day) or unrestricted sleep period group (from 6h/day to 7h/day). Both were 14 to 17 years old. They were instructed to keep the sleep-awakening schedule and sampling protocol. Saliva samples of cortisol and DHEA were collected at 8h, 12h, 16h and 20h. Salivary hormones were analysed with salivary cortisol(or DHEA) EIA kit according to a fixed assay protocol. Cortisol levels of restricted sleep period group and unrestricted sleep period group significantly decreased according to the sampling times. Cortisol levels of sleep restricted group was significantly higher than those of usual sleep group at all sampling times. At 8h, DHEA levels of both groups were significantly higher than those at 12h, 16h and 20h. However, DHEA levels of restricted sleep period group did not differ from those of unrestricted sleep period group at all sampling times.Cortisol and DHEA levels of both group showed the typical diurnal rhythm regardless of sleep status. Restricted sleep may increased cortisol release, not DHEA release, which indicated a changed HPA axis.

Antibacterial and anti-obesity effects of Abeliophyllum distichum Nakai: an in vitro study

  • Song, Dong Cheol;Lee, Ji Hwan;Oh, Han Jin;Kim, Yong Ju;An, Jae Woo;Chang, Se Yeon;Go, Young Bin;Cho, Hyun Ah;Cho, Jin Ho
    • Korean Journal of Agricultural Science
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    • v.48 no.3
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    • pp.557-565
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    • 2021
  • Interest in research on various medicinal plants has increased globally over the last few decades, possibly due to their possible antibacterial and antioxidant activities. The present study was conducted to verify the antioxidant effects, antibacterial activity, and collagen synthesis and cell viability outcomes of adipocytes upon exposure to Abeliophyllum distichum Nakai (AdN). Antibacterial activity was measured through the Disc diffusion method to compare the growth ability of pathogenic microorganisms (E.coli, Salmonella). The absorbance was measured at 560 nm to calculate the active oxygen scavenging ability. Fibroblasts were dispensed in a 96-well plate at a density of 1 × 105 cells·well-1. The amount of procollagen was measured in each case using a procollagen type 1 C-peptide EIA KIT. The cytotoxicity of the Abeliophyllum distichum Nakai extract against animal adipocytes (Hanwoo backfat cells) was determined using a 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay, a method that measures the conversion of MTS to Formazan by means of mitochondrial dehydrogenases. The concentrations of the samples were made to be 0.0125, 0025, 0.05, 0.1, and 0.5% and all were -completely absorbed into the disc in an incubator at 37℃ for 24 to 36 hours. For the 0.125 mg·disc-1, effects of Abeliophyllum distichum Nakai on the antioxidant effect, antibacterial activity, and cell viability of adipocytes were found. However, Abeliophyllum distichum Nakai had no effect on collagen synthesis, thus suggesting that AdN extracts may be useful for the prevention and/or treatment of obesity.

The Assessment of Ultrasensitive HBsAg kit's Sensitivity level and Performance in Detection of Mutant Forms (Ultra-sensitive HBsAg IRMA 키트의 민감도 및 변이형 검출능 평가)

  • Ha, Dong-Hyuk;Min, Kyung-Sun;Noh, Gyeong-Woon;Kim, Hyun-Ju
    • The Korean Journal of Nuclear Medicine Technology
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    • v.15 no.1
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    • pp.121-125
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    • 2011
  • Purpose: The DNA-type virus HBV, discovered by D. Dane and others in 1976, is approximately 42nm big and known as the main cause of liver-related diseases around the world. HBsAg has 4 kinds of subtypes including adw, adr, ayw and ayr and besides common antigen factor a, there are d, y, r, w. From the methods of serologically testing HBV, IRMA, EIA and CLIa were developed for testing HBsAg and are being used in examining the surface antigen of HBV. In this study, among the methods for testing HBV, the recently developed RIAKEY Ultrasensitive HBsAg IRMA kit's sensitivity level and performance in detection of mutant forms were measured and compared with CLIA. Materials and methods: Two certified reference materials, which are WHO 1st International Standard 1985(80/549) and WHO 2nd International Standard 2003(00/588. subtype adw2, genotypeA), were used in the examination and the sensitivity level was measured by diluting these materials from 0.08 IU/ml to 0.005 IU/ml. The materials for examining the detection of mutant forms included 9 kinds of subtype 'ad' and one kind of subtype 'ay' purchased from DSI company. Also, with the use of positive and negative samples, they was compared with CLIA. Result: Ultrasensitive HBsAg kit based on IRMA method showed the detection of up to 0.01 IU/ml not only for WHO 1st International Standard 1985(80/549) but also for WHO 2nd International Standard 2003(00/588. subtype adw2, genotypeA) and the sensitivity level was measured as 0.01 IU/ml by WHO standard. In testing the performance for detection of mutant forms, the 9 kinds of subtype 'ad' and one kind of subtype 'ay' mutant materials were detected, demonstrating the capacity of detecting various types of mutant forms. Conclusions: With the clinical importance of sensitivity level and performance in detection of mutant forms increasing in the field of HBsAg diagnosis, the examination of IRMA's effectiveness using RIA method in the aspects of the sensitivity level and performance in detection of mutant forms was carried out and its result is as follows. The sensitivity level was measured as 0.01 IU/ml by WHO standard and it was possible to measure various types of mutant forms with high sensitivity. Thus it is suggested that more speedy and accurate reports could be produced from a nuclear medicine laboratory for clinical practitioners requiring results of various situations.

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Changes of Measles Specific IgG in the Sera of Children after Birth (출생 후 혈청내 홍역 특히 IgG의 변동)

  • Lee, Kyeong Mee;Yoon, Kang Cheol;Kim, Sun Jun;Cho, Soo Chul;Kim, Jung Soo
    • Pediatric Infection and Vaccine
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    • v.3 no.2
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    • pp.185-193
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    • 1996
  • Purpose : The incidence of measles was gradually decreased since 1963 when measles vaccination was firstly developed and liscenced in the world. But, recently the outbreaks of measles in infants and school children have been reported despite of wide spread use of measles vaccination. This study was performed to evaluate the efficacy of measles vaccination and the necessity of revaccination in Korean infants and children. Methods : 168 subjects of mothers and neonates, infants and children were enrolled in this study during the periods of 10 months from March to December in 1995. Measles specific IgG in the sera of mothers and children was measured using EIA kit (Sigma Co., MO, USA). Antibody titer of over or equal to 110 AU/ml was considered positive. Results : The results obtained were as follows. 1) Values of measles specific IgG in the sera of mother and neonate were 82.9 AU/ml and 89.3 AU/ml respectively and were rapidly decreased within 6 month after birth. Positive antibody levels (${\geq}$ 110 AU/ml) were observed in only 25 % of neonates. 2) In vaccinated children, values of measles specific IgG were 117.4 AU/ml in 9~15 month group, 76.9 AU/ml in 3~6 year group and 79.5 AU/ml in 10~15 year group after either one or two times of measles vaccination. Positive antibody levels in vaccinated children were observed in 57.7% of 9~15 month group, 38.4% of 3~6 year group and 34.7% of 10~15 year group. Conclusion : These results suggest that primary measles vaccination before 6 months of age can be considered and revaccination of measles should be recommended before 3~6 years of age. Further studies will be needed to clarify the reasons of high proportion of primary measles vaccination failure and to established the appropriate schedule of measles vaccination in korean infants and children.

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Identification of proteins involved in the pancreatic exocrine by exogenous ghrelin administration in Sprague-Dawley rats

  • Lee, Kyung-Hoon;Wang, Tao;Jin, Yong-Cheng;Lee, Sang-Bum;Oh, Jin-Ju;Hwang, Jin-Hee;Lim, Ji-Na;Lee, Jae-Sung;Lee, Hong-Gu
    • Journal of Animal Science and Technology
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    • v.56 no.2
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    • pp.6.1-6.4
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    • 2014
  • The aims of study were to investigate the effects of intraperitoneal (i.p.) infusion of ghrelin on pancreatic ${\alpha}$-amylase outputs and the responses of pancreatic proteins to ghrelin that may relate to the pancreatic exocrine. Six male Sprague-Dawley rats (300 g) were randomly divided into two groups, a control group (C, n = 3) and a treatment group (T, $10.0{\mu}g/kg$ BW, n = 3). Blood samples were collected from rat caudal vein once time after one hour injection. The concentrations of plasma ghrelin, cholecystokinin (CCK) and alfa-amylase activity were evaluated by enzyme immunoassay (EIA) kit. Two-dimensional gel electrophoresis (2-DE) analysis was conducted to separate the proteins in pancreas tissue. Results showed that the i.p. infusion of ghrelin at doses of $10.0{\mu}g/kg$ body weight (BW) increased the plasma ghrelin concentrations (p = 0.07) and elevated the plasma CCK level significantly (p < 0.05). Although there was no statistically significant, the ${\alpha}$-amylase activity tended to increase. The proteomics analysis indicated that some pancreatic proteins with various functions were up- or down-regulated compared with control group. In conclusion, ghrelin may have role in the pancreatic exocrine, but the signaling pathway was still not clear. Therefore, much more functional studies focus on these found proteins are needed in the near future.

Anti-inflammatory effect of ethanol extract from Ecklomia cava on gram-positive bacteria(Peptidoglycan)-induced macrophages (그람-양성균(Peptidoglycan)으로 유도한 대식세포에서 감태 에탄올 추출물의 항염증 효과)

  • Kang, Ok-Hwa;Kim, Sung-Bae;Keum, Joon-Ho;Mun, Su-Hyun;Kim, Yong-Sik;An, Byung-Kwan;An, Hyeon-Jin;Kwon, Dong-Yeul
    • Herbal Formula Science
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    • v.19 no.1
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    • pp.195-205
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    • 2011
  • Objectives : Ecklonia cava is brown alga(Laminariaceae) which grows is sea, it has antioxidant, diarrhea and anticoagulant effect. In this study, the effect of ethanol extract of Ecklonia cava (EC) on peptidoglycan(PGN) -induced NO production in RAW 264.7 cells was investigated. Methods : In the present study, IL-6 production was measured using the enzyme-linked immunosorbent assay(ELISA), prostaglandin $\E_2$($\PGE_2$) production was measured using the EIA kit, and inducible NO synthase(iNOS), cyclooxygenase-2(COX-2), and mitogen-activated protein kinase(MAPK) activation, as determined by western blot analysis and reverse transcription -polymerase chain reaction(RT-PCR). Results : EC inhibited PGN-induced NO and IL-6 production. Consistent with these observations, the protein expression of iNOS and COX-2 were inhibited by EC. Moreover, EC suppressed the phosphorylation of extracellular signal-regulated kinase(ERK) 1/2 in PGN-induced RAW 264.7. Conclusions : These results suggest that EC has inhibitory effects on PGN-induced $\PGE_2$, NO, and IL-6 production, as well as the expressions of iNOS and COX-2 in the murine macrophage. These inhibitory effects occur through blockades on the MAPKs phosphorylation.

The study of anti-inflammatory effect of Hyeonto-dan extract in RAW 264.7 macrophage (현토단(玄兎丹)의 RAW 264.7 대식 세포에서의 항염증 효과에 관한 연구)

  • Kim, Ma-Ryong;Kang, Ok-Hua;Kong, Ryong;Seo, Yun-Soo;Zhou, Tian;Kim, Sang-A;Kim, Eun-Su;Sin, Min-A;Lee, Young-Seob;Kwon, Dong-Yeul
    • The Korea Journal of Herbology
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    • v.32 no.2
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    • pp.77-85
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    • 2017
  • Objectives : This study aimed to investigate the unknown mechanisms behind the anti- inflammatory activity of Hyeonto-dan(HT) 70% ethanol extract on LPS-stimulated RAW 264.7 cells. Methods : Cells were treated with Hyeonto-dan 1 h prior to addition of 200 ng/mL of LPS. Cell viability was measured by the MTS assay. Nitric oxide levels were determined by the Griess assay. $PGE_2$ were measured using EIA kit. Pro-inflammatory cytokine production was measured by the enzyme-linked immunosorbent assay (ELISA). The expression of COX-2, iNOS, and MAPKs was investigated by Western blot, qRT-PCR. $NF-{\kappa}B$/p65 localization and interaction of the TLR-4 receptor with LPS was examined by immunofluorescence assays. Results : Hyeonto-dan had no cytotoxicity at the measured concentration. Hyeonto-dan inhibited NO production and pro-inflammatory cytokines such as IL-6, $TNF-{\alpha}$, and PGE2 as well as the protein and mRNA expression of iNOS and COX-2. Moreover, Hyeonto-dan inhibited the interaction between LPS and TLR-4 in murine macrophages. It suppressed phosphorylation of extracellular signal-regulated kinase (ERK 1/2), c-jun N-terminal kinase (JNK 1/2) and p38. Finally, it inhibited translocation of $NF-{\kappa}B$ in response to competitive LPS. Conclusions : Based on the results of this study, Hyeonto-dan inhibited the binding of TLR-4 receptor to LPS and inhibited the phosphorylation of extracellular signaling pathway MAPKs. These inhibitory effects are thought that the amount of $NF-{\kappa}B$ delivered to the nucleus was decreased and the inflammatory reaction was prevented by decreasing the production of LPS-induced $PGE_2$, NO, IL-6 and $TNF-{\alpha}$.

Evaluation of Tuberculosis Activity in Patients with Anthracofibrosis by Use of Serum Levels of IL-2 $sR{\alpha}$, IFN-${\gamma}$ and TBGL(Tuberculous Glycolipid) Antibody (Anthracofibrosis의 결핵활동성 지표로서 혈청 IL-2 $sR{\alpha}$, IFN-${\gamma}$, 그리고 TBGL(tuberculous glycolipid) antibody 측정의 의의)

  • Jeong, Do Young;Cha, Young Joo;Lee, Byoung Jun;Jung, Hye Ryung;Lee, Sang Hun;Shin, Jong Wook;Kim, Jae-Yeol;Park, In Won;Choi, Byoung Whui
    • Tuberculosis and Respiratory Diseases
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    • v.55 no.3
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    • pp.250-256
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    • 2003
  • Background : Anthracofibrosis, a descriptive term for multiple black pigmentation with fibrosis on bronchoscopic examination, has a close relationship with active tuberculosis (TB). However, TB activity is determined in the later stage by the TB culture results in some cases of anthracofibrosis. Therefore, it is necessary to identify early markers of TB activity in anthracofibrosis. There have been several reports investigating the serum levels of IL-2 $sR{\alpha}$, IFN-${\gamma}$ and TBGL antibody for the evaluation of TB activity. In the present study, we tried to measure the above mentioned serologic markers for the evaluation of TB activity in patients with anthracofibrosis. Methods : Anthracofibrosis was defined when there was deep pigmentation (in more than two lobar bronchi) and fibrotic stenosis of the bronchi on bronchoscopic examination. The serum of patients with anthracofibrosis was collected and stored under refrigeration before the start of anti-TB medication. The serum of healthy volunteers (N=16), patients with active TB prior to (N=22), and after (N=13), 6 month-medication was also collected and stored. Serum IL-2 $sR{\alpha}$, IFN-${\gamma}$ were measured with ELISA kit (R&D system, USA) and serum TBGL antibody was measured with TBGL EIA kit (Kyowa Inc, Japan). Results : Serum levels of IL-2 $sR{\alpha}$ in healthy volunteers, active TB patients before and after medication, and patients with anthracofibrosis were $640{\pm}174$, $1,611{\pm}2,423$, $953{\pm}562$, and $863{\pm}401$ pg/ml, respectively. The Serum IFN-${\gamma}$ levels were 0, $8.16{\pm}17.34$, $0.70{\pm}2.53$, and $2.33{\pm}6.67$ pg/ml, and TBGL antibody levels were $0.83{\pm}0.80$, $5.91{\pm}6.71$, $6.86{\pm}6.85$, and $3.22{\pm}2.59$ U/ml, respectively. The serum level of TBGL antibody was lower than of other groups (p<0.05). There was no significant difference of serum IL-2 $sR{\alpha}$ and IFN-${\gamma}$ levels among the four groups. Conclusion : The serum levels of IL-2 $sR{\alpha}$, IFN-${\gamma}$ and TBGL antibody were not useful in the evaluation of TB activity in patients with anthracofibrosis. More useful ways need to be developed for the differentiation of active TB in patients with anthracofibrosis.

Skin Regeneration, Anti-wrinkle, Whitening and Moisturizing Effects of Cheongsangbangpung-tang Aqueous Extracts with Cytotoxicity (청상방풍탕 열수 추출물의 피부재생, 주름개선, 미백, 보습 효과 및 세포독성 평가)

  • Woo, Chang-Yoon;Kim, Dong-Chul
    • The Journal of Korean Obstetrics and Gynecology
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    • v.30 no.2
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    • pp.49-70
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    • 2017
  • Objectives: The present study is to observe the skin-regeneration, anti-wrinkle, whitening and skin moisturizing effects of Cheongsangbangpung-tang (CSBPT) with cytotoxicity. Methods: In the present study, cytotoxicity of CSBPT lyophilized aqueous extracts (yield=18.71%) was experimented against human normal fibroblast cells and B16F10 murine melanoma cells by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium Bromide (MTT) assay, and skin regeneration and anti-wrinkle effects were also showed through the assay of collagen type I synthesis by an enzyme immunoassay (EIA) kit as comparing with transforming growth factor (TGF)-${\beta}1$, hyaluronidase, collagenase and matrix metalloproteinase (MMP)-1 inhibitory assays as comparing with oleanolic acid (OA), and elastase inhibitory effects as comparing with phosphoramidon disodium salt (PP). In addition, whitening effects of CSBPT were observed by tyrosinase inhibitory assay and melanin formation test in B16/F10 melanoma cells as comparing with arbutin, and skin moisturizing effects were measured through mouse skin water contents test, respectively. Results: No CSBPT treatment related cytotoxic effects were demonstrated against human normal fibroblast cells and B16/F10 murine melanoma cells. CSBPT concentration-dependent increased collagen type I synthesis at human normal fibroblast cells. It also effectively suspreessed hyaluronidase, collagenase, elastase and MMP-1 activities, which were enzymes that related to declining of ECM and formation of wrinkle. CSBPT supressed B16/F10 melanoma cells's melanin productions with tyrosinase activity, which was an enzyme connected with melanin formation, and dose-dependent and significant increases of skin water contents were detected in CSBPT treated mouse skin as compared with vehicle control skins. Conclusions: CSBPT showed favorable and enough skin regeneration, anti-wrinkle, whitening and skin moisturizing effects at least in a condition of this experiment. However, more detail mechanism and in vivo skin protective efficacy studies should be conducted in future with the screening of the biological active compounds in individual herbs of Cheongsangbangpung-tang.