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Transforming growth factor-β promoted vascular endothelial growth factor release by human lung fibroblasts (인간 폐섬유아세포에서 TGF-β 자극에 의한 VEGF 분비)

  • Park, Sang-Uk;Shin, Joo-Hwa;Shim, Jae-Won;Kim, Deok-Soo;Jung, Hye-Lim;Park, Moon-Soo;Shim, Jung-Yeon
    • Clinical and Experimental Pediatrics
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    • v.51 no.8
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    • pp.879-885
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    • 2008
  • Purpose : The human lung fibroblast may act as an immunomodulatory cell by providing pro-inflammatory cytokines and chemokines, which are important in airway remodeling. Vascular endothelial growth factor (VEGF) induces mucosal edema and angiogenesis. Thymus and activation regulated chemokine (TARC) induces selective migration of T helper 2 cells. We investigated whether human lung fibroblasts produced VEGF and TARC, and the effects were augmented with the co-culture of fibroblasts and human bronchial smooth muscle cells (HBSMC), and whether dexamethasone can inhibit the proliferation and the release of VEGF in lung fibroblasts. Methods : Human lung fibroblasts were cultured with and without HBSMC, growth-arrested in serum-deprived medium, and pretreated with dexamethasone for 16 hours. After 24-hour stimulation with platelet derived growth factor-BB (PDGF-BB) and/or transforming growth factor-${\beta}$ (TGF-${\beta}$), culture supernatant was harvested for assays of VEGF and TARC. Cell proliferation was assayed using BrdU cell proliferation ELISA kit. Results : 1) The release of VEGF was significantly increased after stimulation with TGF-${\beta}$, and its release was augmented when co-stimulated with PDGF and TGF-${\beta}$. 2) VEGF release induced by PDGF or TGF-${\beta}$ was inhibited by dexamethasone. 3) There was no synergistic effect on the release of VEGF when human lung fibroblasts were co-cultured with HBSMC. 4) Dexamethasone did not suppress human lung fibroblasts proliferations. 5) Neither TGF-${\beta}$ nor PDGF induced TARC release from lung fibroblasts. Conclusion : Human lung fibroblasts may modulate airway remodeling by release of VEGF, but they have no synergistic effects when co-cultured with HBSMC. Dexamethasone suppresses VEGF release, not proliferation of lung fibroblast.

Effects of Heat-stress on Rumen Bacterial Diversity and Composition of Holstein Cows (고온 스트레스 영향에 따른 홀스타인종 젖소의 반추위내 미생물 균총 변화)

  • Kim, Dong Hyeon;Kim, Myung Hoo;Kim, Sang Bum;Ha, Seung Min;Son, Jun Kyu;Lee, Ji Hwan;Hur, Tai Young;Lee, Jae Yeong;Park, Ji Hoo;Choi, Hee Chul;Lee, Hyun Jeong;Park, Beom Young;Ki, Kwang Seok;Kim, Eun Tae
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.39 no.4
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    • pp.227-234
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    • 2019
  • This study was performed to investigate the effect of heat-stressed environment on rumen microbial diversity in Holstein cows. Rectal temperature and respiration rate were measured and rumen fluid was collected under normal environment (NE; Temperature humidity index (THI)=64.6) and heat-stressed environment (HE; THI=87.2) from 10 Holstein cows (60±17.7 months, 717±64.4 kg) fed on the basis of dairy feeding management in National Institute of Animal Science. The rumen bacteria diversity was analyzed by using the Illumina HiSeqTM 4000 platform. The rectal temperature and respiratory rate were increased by 1.5℃ and 53 breaths/min in HE compared to that in NE, respectively. In this study, HE exposure induced significant changes of ruminal microbe. At phylum level, Fibrobacteres were increased in HE. At genus level, Ruminococcaceae bacterium P7 and YAD3003, Butyrivibrio sp. AE2032, Erysipelotrichaceae bacterium NK3D112, Bifidobacterium pseudolongum, Lachnospiraceae bacterium FE2018, XBB2008, and AC2029, Eubacterium celulosolvens, Clostridium hathewayi, and Butyrivibrio hungatei were decreased in HE, while Choristoneura murinana nucleopolyhedrovirus, Calothrix parasitica, Nostoc sp. KVJ20, Anabaena sp. ATCC 33047, Fibrobacter sp. UWB13 and sp. UWB5, Lachnospiraceae bacterium G41, and Xanthomonas arboricola were increased in HE. In conclusion, HE might have an effect to change the rumen microbial community in Holstein cows.

Detection of Point Mutations in the rpoB Gene Related to Drug Susceptibility in Mycobacterium Tuberculosis using an Oligonucleotide Chip (올리고뉴클레오티드 칩(Oligonucleotide Chip)을 이용한 항결핵제 감수성과 관련된 Mycobacterium tuberculosis rpoB 유전자의 점돌연변이 판별 방법)

  • Kim, Hyun-Jung;Kim, Seong-Keun;Shim, Tae-Sun;Park, Yong-Doo;Park, Mi-Sun
    • Tuberculosis and Respiratory Diseases
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    • v.50 no.1
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    • pp.29-41
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    • 2001
  • Background : The appearance of multiple-drug-resistant Mycobacterium tuberculosis strains has been seriously compromising successful control of tuberculosis. Rifampin-resistance, caused by mutations in the rpoB gene, can be indicative of multiple-drug-resistance, and its detection is of great importance. The present study aimed to develop an oligonucleotide chip for accurate and convenient screening of drug-resistance. Methods : In order to detect point mutations in the rpoB gene, an oligonucleotide chip was prepared by immobilizing specific probe DNA to a microscopic slide glass by a chemical reaction. The probe DNA that was selected from the 81 bp core region of the rpoB gene was designed to have mutation sites at the center. A total of 17 mutant probes related to rifampin-resistance including 8 rifabutin-sensitive mutant probes were used in this study. For accurate determination, wild type probes were prepared for each mutation position with an equal length, which enabled a direct comparison of the hybridization intensities between the mutant and wild type. Results : Mycobacterial genomic DNA from clinical samples was tested with the oligonucleotide chip and the results were compared with those of the drug-susceptibility test in addition to sequencing and INNO-LiPA Rif. TB kit test in some cases. Out of 15 samples, the oligonucleotide chip results of 13 samples showed good agreement with the rifabutin-sensitivity results. The two samples with conflicting result also showed a discrepancy between the other tests, suggesting such possibilities as existence of mixed strains and difference in drug-sensitivity. Further verification of these samples in addition to more case studies are required before the final evaluation of the oligonucleotide chip can be made. Conlcusion : An oligonucleotide chip was developed for the detection of rpoB gene mutations related to drugsusceptibility. The results to date show the potential for using the oligonucleotide chip for accurate and convenient screening of drug-resistance to provide useful information in antituberculosis drug therapy.

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Protective Effect of Radiation-induced New Blackberry Mutant γ-B201 on H2O2-induced Oxidative Damage in HepG2 Cells (H2O2 에 의해 유도된 HepG2 세포의 산화적 스트레스에 대한 신품종 방사선 돌연변이 블랙베리 γ-B201의 세포 보호 효과)

  • Cho, Byoung Ok;Lee, Chang-Wook;So, Yangkang;Jin, Chang-Hyun;Yook, Hong-Sun;Byun, Myung-Woo;Jeong, Yong-Wook;Park, Jong Chun;Jeong, Il-Yun
    • Korean Journal of Food Science and Technology
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    • v.46 no.3
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    • pp.384-389
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    • 2014
  • The objective of the present study was to investigate the chemical composition of anthocyanin-enriched extract of radiation-induced blackberry (Rubus fruticosus L.) mutant (${\gamma}$-B201) as well as the protective effect of ${\gamma}$-B201 against oxidative stress in vitro. The cytotoxicity, reactive oxygen species (ROS) scavenging capacity, and DNA damage were assessed by WST-1 assay, flow cytometry, and comet assay, respectively. Lactate dehydrogenase, superoxide dismutase, and catalase activities were determined by using a commercial kit. The in vitro results showed that ${\gamma}$-B201 increased the cell viability, reduction of lactate dehydrogenase release, and intracellular ROS scavenging capacity in hydrogen peroxide ($H_2O_2$)-treated HepG2 cells. Furthermore, treatment with ${\gamma}$-B201 attenuated DNA damage in $H_2O_2$-treated HepG2 cells and treatment with ${\gamma}$-B201 restored the activity of superoxide dismutase and catalase in $H_2O_2$-treated HepG2 cells. In conclusion, the present study suggests that ${\gamma}$-B201 blackberry extract can exert a significant cytoprotective effect against $H_2O_2$-induced cell damage.

Splenocyte-mediated immune enhancing activity of Sargassum horneri extracts (괭생이 모자반 추출물의 비장세포 면역활성 증강 효과)

  • Kim, Dong-Sub;Sung, Nak-Yun;Han, In-Jun;Lee, Byung-Soo;Park, Sang-Yun;Nho, Eun Young;Eom, Ji;Kim, Geon;Kim, Kyung-Ah
    • Journal of Nutrition and Health
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    • v.52 no.6
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    • pp.515-528
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    • 2019
  • Purpose: This study examined the immunological activity and optimized the mixture conditions of Sargassum horneri (S. horneri) extracts in vitro and in vivo models. Methods: S. horneri was extracted using three different methods: hot water extraction (HWE), 50% ethanol extraction (EE), and supercritical fluid extraction (SFE). Splenocyte proliferation and cytokine production (Interleukin-2 and Interferon-γ) were measured using a WST-1 assay and enzyme-linked immunosorbent assay, respectively. The levels of nitric oxide and T cell activation production were measured using a Griess assay and flow cytometry, respectively. The natural killer (NK) cell activity was determined using an EZ-LDH kit. Results: Among the three different types of extracts, HWE showed the highest levels of splenocyte proliferation and cytokine production in vitro. In the animal model, three different types of extracts were administrated for 14 days (once/day) at 50 and 100 mg/kg body weight. HWE and SFE showed a high level of splenocyte proliferation and cytokine production in the with and without mitogen-treated groups, whereas EE administration did not induce the splenocyte activation. When RAW264.7 macrophage cells were treated with different mixtures (HWE with 5, 10, 15, 20% of SFE) to determine the optimal mixture ratio of HWE and SFE, the levels of nitric oxide and cytokine production increased strongly in the HWE with 5% and 10% of SFE containing group. In the animal model, HWE with 5% and 10% of SFE mixture administration increased the levels of splenocyte proliferation, cytokine production, and activated CD4+ cell population significantly, with the highest level observed in the HWE with 5% of SFE group. Moreover, the NK cell activity was increased significantly in the HWE with 5% of SFE mixture-treated group compared to the control group. Conclusion: The optimal mixture condition of S. horneri with immune-enhancing activity is the HWE with 5% of SFE mixture. These results confirmed that the extracts of S. horneri and its mixtures are potential candidate materials for immune enhancement.

A Comparative Study of Production of [68Ga]PSMA-11 with or without Cassette Type Modules (비 카세트 방식과 카세트 방식을 이용한 [68Ga]PSMA-11의 자동 합성 방법 비교)

  • Hyun-Sik, Park;Byeong-Min, Jo;Hyun-Ho, An;Hong-Jin, Lee;Jin-Hyeong, Lee;Gyeong-Jae, Lee;Byung-Chul, Lee;Won-Woo, Lee
    • The Korean Journal of Nuclear Medicine Technology
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    • v.26 no.2
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    • pp.15-19
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    • 2022
  • Purpose [68Ga]PSMA-11 is needed the high reproducibility, excellent radiochemical yield and purity. In term of radiation safety, the radiation exposure of operator for its production also should be considered. In this work, we performed a comparative study for the fully automated synthesis of [68Ga]PSMA-11 between non-cassette type and cassette type. Materials and Methods Two different type of modules (TRACERlab FX N pro for non-cassette type and BIKBox for cassette type) were used for the automated production of [68Ga]PSMA-11. According to the previously identified elution profile, Only 2.5 ml with high radioactivity was used for the reaction. After adjusting the pH of the reaction solution with HEPES buffer solution, the precursor was added and reacted with at 95 ℃ for 15 minutes. The reaction mixture was separated and purified using a C18 light cartridge. The product was eluted with 50% EtOH/saline solution and diluted with saline. It was completed by sterilizing filter. In the non-cassette type, the aforementioned process must be prepared directly. However, in the cassette method, synthesis was possible simply by installing a kit that was already completed. Results Both total [68Ga]PSMA-11 production time were 25±3(non-cassette type) and 23±3 minutes(cassette type). The radiochemical yield of the non-cassette type(65.5±5.7%) was higher than that of the cassette type(61.6±4.8%) after sterilization filter. The non-cassette type took about 120 minutes of preparation time before synthesis due to washing of synthesizer and reagent preparation. However, since the cassette type does not require washing and reagent preparation, it took about 20 minutes to prepare before synthesis. Both type of synthesizer had a radiochemical high purity(>99%). Conclusion The non-cassette type production of [68Ga]PSMA-11 showed higher radiochemical yield and lower cost than the cassette type. However, The cassette type has an advantage in terms of preparation time, convenience, and equipment maintenance.

Pipetting Stability and Improvement Test of the Robotic Liquid Handling System Depending on Types of Liquid (용액에 따른 자동분주기의 분주능력 평가와 분주력 향상 실험)

  • Back, Hyangmi;Kim, Youngsan;Yun, Sunhee;Heo, Uisung;Kim, Hosin;Ryu, Hyeonggi;Lee, Guiwon
    • The Korean Journal of Nuclear Medicine Technology
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    • v.20 no.2
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    • pp.62-68
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    • 2016
  • Purpose In a cyclosporine experiment using a robotic liquid handing system has found a deviation of its standard curve and low reproducibility of patients's results. The difference of the test is that methanol is mixed with samples and the extractions are used for the test. Therefore, we assumed that the abnormal test results came from using methanol and conducted this test. In a manual of a robotic liquid handling system mentions that we can choose several setting parameters depending on the viscosity of the liquids being used, the size of the sampling tips and the motor speeds that you elect to use but there's no exact order. This study was undertaken to confirm pipetting ability depending on types of liquids and investigate proper setting parameters for the optimum dispensing ability. Materials and Methods 4types of liquids(water, serum, methanol, PEG 6000(25%)) and $TSH^{125}I$ tracer(515 kBq) are used to confirm pipetting ability. 29 specimens for Cyclosporine test are used to compare results. Prepare 8 plastic tubes for each of the liquids and with multi pipette $400{\mu}l$ of each liquid is dispensed to 8 tubes and $100{\mu}l$ of $TSH^{125}I$ tracer are dispensed to all of the tubes. From the prepared samples, $100{\mu}l$ of liquids are dispensed using a robotic liquid handing system, counted and calculated its CV(%) depending on types of liquids. And then by adjusting several setting parameters(air gap, dispense time, delay time) the change of the CV(%)are calcutated and finds optimum setting parameters. 29 specimens are tested with 3 methods. The first(A) is manual method and the second(B) is used robotic liquid handling system with existing parameters. The third(C) is used robotic liquid handling system with adjusted parameters. Pipetting ability depending on types of liquids is assessed with CV(%). On the basis of (A), patients's test results are compared (A)and(B), (A)and(C) and they are assessed with %RE(%Relative error) and %Diff(%Difference). Results The CV(%) of the CPM depending on liquid types were water 0.88, serum 0.95, methanol 10.22 and PEG 0.68. As expected dispensing of methanol using a liquid handling system was the problem and others were good. The methanol's dispensing were conducted by adjusting several setting parameters. When transport air gap 0 was adjusted to 2 and 5, CV(%) were 20.16, 12.54 and when system air gap 0 was adjusted to 2 and 5, CV(%) were 8.94, 1.36. When adjusted to system air gap 2, transport air gap 2 was 12.96 and adjusted to system air gap 5, Transport air gap 5 was 1.33. When dispense speed was adjusted 300 to 100, CV(%) was 13.32 and when dispense delay was adjusted 200 to 100 was 13.55. When compared (B) to (A), the result increased 99.44% and %RE was 93.59%. When compared (C-system air gap was adjusted 0 to 5) to (A), the result increased 6.75% and %RE was 5.10%. Conclusion Adjusting speed and delay time of aspiration and dispense was meaningless but changing system air gap was effective. By adjusting several parameters proper value was found and it affected the practical result of the experiment. To optimize the system active efforts are needed through the test and in case of dispensing new types of liquids proper test is required to check the liquid is suitable for using the equipment.

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A Survey on the Perception of the Counterplans of Medical Accident and Dispute of Dental Hygienist (의료사고 및 의료분쟁에 대한 치위생사의 인식도 조사)

  • Oh, Jin-Ho;Kwon, Jeong-Seung;Ahn, Hyoung-Joon;Kang, Jin-Kyu;Choi, Jong-Hoon
    • Journal of Oral Medicine and Pain
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    • v.32 no.1
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    • pp.9-33
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    • 2007
  • In the field of dentistry, there existed relatively few emergency patients or patients who need intensive care and thus had low medical dispute rates. However, these days, there is a general tendency of increased medical disputes. Although many medical disputes are caused by medical accidents of the dentists, because dental assistants are also lawfully involved in practicing dentistry, there is a possibility of medical disputes or medical accidents caused by dental assistants. Therefore, the role of the dental assistants cannot be ignored. This study consists of a survey given to dental hygienists currently working in general hospitals, dental hospitals and private dental clinics. Following is the results of the analysis of 275 respondents' backgrounds, medical disputes rates including patients' complaints, their understanding of medical regulations and their general understanding of overall dental practice and medical disputes. 1. 251 of 274(91.6%) respondents doubted the risk of medical accident and dispute. 2. 81(29.5%) dental hygienist experienced complaint from patients. They have been working in the private dental clinic, the rate of this experience was high. 3. 349 case of 1805(19.3%) the complaints by patients, highest percentage among its category, were those regarding dental fees and poor service. 4. 129 case of 1805(7.1%) patients' complaints, highest percentage among it's subcategory, were those regarding the absence of explanations of precautions or request of agreements before dental treatment. 5. 252 of 267 (94.4%) dental hygienists chart after a scaling treatment. However, only 55(20.7%) dental hygienists chart the fact of explaining the precautions. 6. 6(2.2%) dental hygienists do not inspect patients' medical history, if patients don't mention it. 7. 104 of 274(38.0%) dental hygienists responded to be capable of administering first aid treatment. 8. 115(41.8%) dental hygienists have a first aid kit and equipment. 9. In case of medical dispute, 268(97.8%) dental hygienists respond that, charting plays a big role in resolving the dispute. 10. In case of medical dispute, 272(93.3%) dental hygienists respond that, explanation and agreement before treatment have an important role in settlement of dispute 11. Only 160(58.4%) dental hygienists responded correct answer that the duration of keeping medical records is 10 years. 12. 124(45.3%) respondents thought that it is legal for a dental hygienist to take a panoramic dental X-ray, 71(25.9%) respondents thought that it is legal practice cervical resin treatment by dental hygienist, and 37(13.5%) respondents thought that it is legal extract primary teeth by dental hygienist. 13. 24(18.76%) respondents thought that it doesn't matter to tell patient's state to others 14. 272(99.27%) responded that receiving education for the prevention of medical disputes was needed and of them, 61.0% thought it was urgent. 15. 186(64.2%) has never had classes regarding the prevention of medical disputes while in school and 212(77.4%) has not had the same type of classes after graduating from school. 16. 256(93.4%) responded that there will be even more of an increased number of medical disputes. Among them, 83.3% of respondents though that due to the increased opportunity of acquiring information through the internet and mass media. The study shows that 29.5 percentage of dental hygienists have experienced the medical disputes and complaints and they are lack of recognition of medical regulations and dental hygienist's official duty. So, there is a big potential of the percentage to increase. Therefore, the correct understanding of explaining precautions and requesting agreement before dental treatments and performing them are mandatory. Moreover, classes regarding the prevention and counterplans of medical disputes need to be widely offered.

Oestrogenic Activity of Parabens In Vitro Estrogen Assays (에틸, 프로필, 이소프로필, 부틸, 이소부틸 파라벤의 In Vitro 검색시험 연구에서의 내분비독성)

  • Lee Sung-Hoon;Kim Sun-Jung;Park Jung-Ran;Jo Eun-Hye;Ahn Nam-Shik;Park Joon-Suk;Hwang Jae-Woong;Jung Ji-Youn;Lee Yong-Soon;Kang Kyung-Sun
    • Journal of Food Hygiene and Safety
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    • v.21 no.2
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    • pp.100-106
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    • 2006
  • The use of underarm and body care cosmetics with oestrogenic chemical excipients (particularly the parabens) and the hypothesized association with breast cancer incidence, particularly in women. It is noted that the type of cosmetic product is irrelevant (e.g. antiperspirant/deodorant versus body lotion, moisturizers or sprays versus creams) and attention must focus on issues of actual exposure to chemicals through continued dermal application of body care products and the endocrine/hormonal activity and toxicity of the chemicals in the formulations. To evaluate the estrogenic activities of parabens such as ethylparaben, butylparaben, propylparaben, isobutylparaben and isopropylparaben, we used recombinant yeasts containing the human estrogen receptor [Saccharomyces cerevisiae ER+LYS 8127], human breast cancer MCF-7 cell lines and human estrogen receptor ${\alpha}\;and\;{\beta}$. In E-screen assays, isopropylparaben is the most estrogenic paraben, and in ER competition assay, isobutylparaben is the most estrogenic paraben. We evaluated isopropylparaben was most active in the recombinant yeast assay, followed by propylparaben, ethylparaben, isobutylparaben and butylparaben. Results from this study demonstrate that parabens are observed in human endocrine system. Therefore, we have shown that the parabens is induced the estrogenic activities similar to $17{\beta}$-estradiol and Bisphenol-A.