• Title/Summary/Keyword: Cell-based assay

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Effect of Whalakyuoleyng-dan plus Yinsamyangwui-tang on Anti-angionesis (활락효영단합인삼양위탕(活絡效靈丹合人蔘養胃湯)이 혈관신생(血管新生) 억제(抑制)에 미치는 영향(影響))

  • Ko, Ki-Wan;Park, Joon-Hyuk;Kang, Hee;Kim, Sung-Hoon;Yu, Young-Beob;Shim, Bum-Sang;Choi, Seung-Hoon;Ahn, Koo-Seok
    • THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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    • v.7 no.1
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    • pp.77-97
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    • 2001
  • Anti-angiogenesis is one of therapies which have been high-lightened on the research of cancer treatment. Anti-angiogenesis means that new blood vessels are created from a existing capillary tube and it is a important process on metastasis and permeation when cancer is created or formed. Since angiogenesis have been under research, a complete recovery oriented treatment against cancer have been suggested blocking metastasis, delaying the growth of cancer cell, and blocking the supply of oxygen and nutritive substance through the web of blood vessels. Until now, there are several anti-angiogenesis, which have been known to the public, such as thalidomide, angiostatin, endostatin, 2-methoxyestradiol, TNP-470, and marimastat, etc. Additionally, 17 clinical testing projects about anti-angiogenesis are on the process in NCI(National Cancer Institute). Especially, TNP-470 showed effectiveness against cancer on clinical testing after finishing animal testing. Based on existing researches showing that Yinsamyangwui-tang is effective to strengthening body resistance and Whallakhyolenyng-dan effects cells on the inside of blood vessel because Whallakhyolenyng- dan restrains cell adhesion during the restraining period of a blood vessel, I tried to research the effect of Whalakhyolenyng-dan plus Yinsamyangwui-tang on angiogenesis. I made a conclusion putting into operation through using SK-Hep-1 (KCLB 30052), A549(KCLB 10185), AGS(KCLB 21739), and BCE(Bovine Capillary Endothelial Cell). Followings are the results of my experimental research: 1. According to the researching results of anti-cancer activation against cancer cell, Whallkhyoleyng dan plus Yinsamyangwui-tang decreased the number of cancer cells -- While injecting $600{\mu}g/ml$, injected groups decreased 3.1% more comparing with the contrastive group of SK-Hep-1, 49.7% more comparing with the contrastive group of A549, and 31.0% more comparing with the contrastive group of AGS. 2. According to the researching results of DNA composition effect between BCE and cancer cell, Whallakhyoleyng-dan plus Yinsamyangwui-tang reduced the rate of SK-Hep-1 synthesis inhibition by 59.1% at $600{\mu}g/ml$ intensity comparing with contrastive group; for A549, 72.6%; for AGS, 6.1%, for BCE, 28.9%. 3. According to the researching results about the effect of BCE cell to angiogenesis, angiogenesis was restrained at $400{\mu}g/ml$ intensity during 18 hours observation. 4. In the case of aortic ring assay, the half level of angiogenesis was reduced comparing with the contrastive group while injecting with $400{\mu}g/ml$ intensity; with $800{\mu}g/ml$, under 10% comparing with contrastive group; and with $1600{\mu}g/ml$, complete restrain. According to the above results, Whallakhyoleyng-dan plus Yinsamyangwui-tang was proved to have an anti-angiogenetic effects.

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Effects of Vegetable Peptones on Promotion of Cell Proliferation and Collagen Production (Vegetable Peptones의 세포증식 및 콜라겐생성 촉진효과)

  • Jung, Eun-Sun;Lee, Jong-Sung;Lee, Jienny;Huh, Sung-Ran;Kim, Young-Soo;Hwang, Wang-Taek;Park, Deok-Hoon
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.35 no.1
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    • pp.65-72
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    • 2009
  • Skin aging appears to be principally attributed to a decrease in both levels of Type I collagen and regeneration ability of dermal fibroblasts. It is important to introduce an efficient and safe agent for effective management of skin aging. To this end, we performed screening for anti-ageing agents and then found that vegetable peptones (pea and wheat) promoted cell proliferation of adult stem cells. Vegetable peptones may be considered as useful medium additives because it can supply nutrients, peptides, amino acids or growth factor analogues. This study was designed to investigate effects of vegetable peptones on cell proliferation/collagen production and their possible mechanisms in human dermal fibroblasts. In cell proliferation assay, vegetable peptones significantly promoted cell proliferation in a concentration-dependent manner. In addition, human COL1A2 promoter luciferase and type I procollagen synthesis assays showed that vegetable peptones induce type I procollagen production through the activation of COLlA2 promoter. In both TGF-${\beta}1$ luciferase reporter and ELISA assays, vegetable peptones was found to induce TGF-${\beta}1$ production, suggesting that vegetable peptones induce type I procollagen production through the activation of TGF-${\beta}1$. When applied topically in a human skin twice a day for an 4-week period of time, vegetable peptones did not induce any adverse reactions. Theretore, based on these results, we suggest the possibility that vegetable peptones may be considered as an attractive, wrinkle-reducing candidate for topical application.

Development and Application of a Novel Mammalian Cell Culture System for the Biocompatibility and Toxicity of Polymer Films and Metal Plate Biomaterials (고분자필름과 금속막 의료소재에 대한 생체적합성 및 독성 평가를 위한 새로운 세포배양시스템의 개발 및 적용)

  • Kwak, Moon Hwa;Yun, Woo Bin;Kim, Ji Eun;Sung, Ji Eun;Lee, Hyun Ah;Seo, Eun Ji;Nam, Gug Il;Jung, Young Jin;Hwang, Dae Youn
    • Journal of Life Science
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    • v.26 no.6
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    • pp.633-639
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    • 2016
  • Biomaterials including polymer, metal, ceramic, and composite have been widely applied for medical uses as medical fibers, artificial blood vessels, artificial joints, implants, soft tissue, and plastic surgery materials owing to their physicochemical properties. However, the biocompatibility and toxicity for film- and plate-form biomaterials is difficult to measure in mammalian cells because there is no appropriate incubation system. To solve these problems, we developed a novel mammalian cell culture system consisting of a silicone ring, top panel, and bottom panel and we applied two polymer films (PF) and one metal plate (MP). This system was based on the principal of sandwiching a test sample between the top panel and the bottom panel. Following the assembly of the culture system, SK-MEL-2 cells were seeded onto Styela Clava Tunic (SCT)-PF, NaHCO3-added SCT (SCTN)-PF, and magnesium MP (MMP) and incubated at 37℃ for 24 hr and 48 hr. An MTT assay revealed that cell viability was maintained at a normal level in the SCT-PF culture group at 24 or 48 hr, although it rapidly decreased in the SCTN-PF culture group at 48 hr. Furthermore, the cell viability in the MMP culture group was very similar to that of the control group after incubation for 24 hr and 48 hr. Together, these results suggest the sandwich-type mammalian culture system developed here has the potential for the evaluation of the biocompatibility and toxicity of cells against PF- and MP-form biomaterials.

Anti-Inflammatory Effect of Ethanol Extract from Grateloupia elliptica Holmes on Lipopolysaccharide-Induced Inflammatory Responses in RAW 264.7 Cells and Mice Ears (LPS로 유도된 RAW 264.7 세포와 마우스 귀 조직에 대한 참도박(Grateloupia elliptica Holmes) 에탄올 추출물의 항염증 효과)

  • Bae, Nan-Young;Kim, Min-Ji;Kim, Koth-Bong-Woo-Ri;Ahn, Na-Kyung;Choi, Yeon-Uk;Park, Ji-Hye;Park, Sun-Hee;Ahn, Dong-Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.8
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    • pp.1128-1136
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    • 2015
  • This study investigated the anti-inflammatory effects of ethanol extract from Grateloupia elliptica Holmes (GEHEE) on the lipopolysaccharide-induced inflammatory response. Anti-inflammatory effects were detected by enzyme-linked immunosorbent assay, Western blotting, and immunohistochemistry. There were no cytotoxic effects on proliferation of macrophages treated with GEHEE compared to the control. GEHEE remarkably suppressed NO and pro-inflammatory cytokines (interleukin-6, tumor necrosis $factor-{\alpha}$, and $interleukin-1{\beta}$) production and reduced expression of inducible nitric oxide synthase, cyclooxygenase-2, and nuclear $factor-{\kappa}B$ ($NF-{\kappa}B$) proteins in a dose-dependent manner. GEHEE also significantly reduced activation of mitogen-activated protein kinases (MAPKs). The formation of edema in mouse ears was reduced at the highest dose compared to the control. GEHEE also reduced dermal thickness and mast cell numbers based on histological analysis. These results suggest that GEHEE exerts significant anti-inflammatory activity via inhibition of $NF-{\kappa}B$ and MAPKs activation and may be a potential anti-inflammatory therapeutic material.

Increased Regulatory T cells in Thyroid Adenocarcinoma in a Mixed Breed Dog (갑상샘 암종을 가진 개에서 조절 T 세포의 증가)

  • Kim, Jun-Hwan;Hong, Yeon-Jung;Kim, Bumseok;Kim, Yunhye;Park, Jin-Ho;Chung, Tae-Ho;Park, Chul
    • Journal of Veterinary Clinics
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    • v.30 no.6
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    • pp.482-485
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    • 2013
  • This case report describes the increment of regulatory T cells in thyroid carcinoma of a mixed breed dog compared with normal dogs. A mixed breed dog was referred for a submandibular mass and hyperthermia. Right cervical mass was detected during the physical examination and radiography identified right cervical mass suspected thyroid gland. Accurate location and size of mass was confirmed using computed tomography and 3D reconstructed images. After confirmation of mass, surgical resection was performed and evaluation of regulatory T cells in blood of this patient was performed using flow cytometric assay. The percentage of regulatory T cells was 38.28% of all CD4 (+)/CD25 (+) T cells. This result was higher than median percentage of regulatory T cells of healthy 8 beagles ($7.66{\pm}1.65%$ (p<0.01) of all CD4 (+)/CD25 (+) T cells). And masses were confirmed as giant cell thyroid carcinoma based on histopathologic examination. After surgical resection of cervical mass was performed, the owner didn't want chemotherapy. Seven days later, cervical mass came out again. And CBC showed severe leukocytosis (WBC $47.6{\times}10^3/{\mu}L$) and non-regenerative anemia (PCV 21%). Suspected pulmonic metastasis regions became more severe. Finally the dog died of severe anemia and respiratory disorder.

Multi-Parameter Approach for Evaluation of Genomic Instability in the Polycystic Ovary Syndrome

  • Sekar, Nishu;Nair, Manju;Francis, Glory;Kongath, Parvathy Raj;Babu, Sandhya;Raja, Sudhakaran;Gopalakrishnan, Abilash Valsala
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.16
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    • pp.7129-7138
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    • 2015
  • Background: The polycystic ovary syndrome (PCOS), characterized by hyperandrogenism and chronic anovulation, is a common endocrine disorder in women. PCOS, which is associated with polycystic ovaries, hirsutism, obesity and insulin resistance, is a leading cause of female infertility. In this condition there is an imbalance in female sex hormones. All the sequelae symptoms of PCOS gradually lead to cancer in the course of time. It is heterogeneous disorder of unknown etiology so it is essential to find the exact cause. Materials and Methods: In this study both invasive and non-invasive techniques were employed to establish the etiology. Diagnosis was based on Rotterdam criteria (hyperandrogenism, ovulatory dysfunction, PCOM) and multiparameters using buccal samples and dermatoglypic analysis and cytogenetic study for 10 cases and four age and sex matched controls. Results: In clinical analysis we have observed the mean value of total testosterone level was 23.6nmol/L, total hirsutism score was from 12-24, facial acne was found in in 70% patients with 7-12 subcapsular follicular cysts, each measuring 2-8 mm in diameter. In dermatoglypic analysis we observed increases in mean value ($45.9^{\circ}$) of ATD angle when compared with control group and also found increased frequency (38%) of Ulnar loops on both fingers (UU), (18%) whorls on the right finger and Ulnar loop on left finger (WU) and (16%) arches on right and left fingers (AA) were observed in PCOS patients when compared with control subjects. Features which could be applied as markers for PCOS patients are the presence of Ulnar loops in middle and little fingers of right and left hand. The buccal micronucleus cytome assay in exfoliated buccal cells, we found decrease in frequency of micronuclei and significant increases in frequency of karyolysed nuclei in polycystic ovarian syndrome patients. Chromosome aberration analysis revealed a significant increase in frequency of chromosome aberrations (CAs) in PCOS patients when compared with controls. Conclusions: From this present work it can be concluded that non-invasive technique like dermatoglypics analysis and buccal micronucleus cytome assays with exfoliated buccal cell can also be effective biomarkers for PCOS, along with increased CAs in lymphocytes as a sign of genetic instability. There is a hypothesis that micronuclei and chromosomal aberrations could have a predictive value for cancer. From this present work it can be concluded to some extent that non-invasive technique like dermatoglypics and buccal cell analysis can also be effective for diagnosis.

Hazard Assessment of Green-Wall Plant Campsis grandiflora K. Schum in Urban Areas based on Pollen Morphology and Cytotoxicity (도심 벽면녹화식물 능소화 (Campsis grandiflora K. Schum)의 화분 형태 및 세포독성에 근거한 유해성평가)

  • Kim, Hyun-Jun;So, Soonku;Shin, Chang-Ho;Noh, Hae-ji;Na, Chun-Soo;Lee, You-Mi
    • Korean Journal of Environmental Biology
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    • v.33 no.2
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    • pp.256-261
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    • 2015
  • This study was conducted to assess the influence of hazard pattern in the surface structure of pollen grains of Campsis grandiflora, and cytotoxicity of different part extracts and nectar on RAW264.7 macrophages. The pollen grains were medium sized ($21.8{\mu}m$) with tricolpate aperture type. In equatorial view, the pollens were prolate (P/E=1.8) and the exine pattern was smooth and reticulate. This result contradict with the rumor of having a hook-shaped protuberance that can damage the cornea because we couldn't observed any protuberance on the surface of the outer wall. Furthermore, we investigated the 70% MeOH extracts (flower, leaf, stem) and nectar of C. grandiflora for their cell viability in temporal basis via MTT analysis on RAW264.7 macrophage cells. There was no significant difference in the cytotoxicity among the MeOH extracts and nectar of C. grandiflora after 24 h. However, nectar showed the dosedependent cytotoxicity on RAW264.7 macrophage cells after 48 h.

Distribution of Oncogenic Human Papillomavirus Genotypes at High Grade Cervical Lesions above CIN 2 Grade with Histological Diagnosis

  • Kim, Geehyuk;Park, Sungyoung;Wang, Hye-young;Kim, Sunghyun;Park, Sangjung;Yu, Kwangmin;Lee, Boohyung;Ahn, Seung-Ju;Kim, Eun-Joong;Lee, Dongsup
    • Biomedical Science Letters
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    • v.22 no.2
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    • pp.37-45
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    • 2016
  • High risk human papillomavirus (HR-HPV) is major risk factor for uterine cervical cancer. There are approximately 15 types of HR-HPV. Liquid based cytology samples (116 samples) with high grade cervical lesions belonging to cervical intraepithelial neoplasia (CIN) 2, CIN 3, carcinoma in situ (CIS) and squamous cell carcinoma (SCC) were used after histologic confirmation. HR-HPV genotype assay was conducted using DNA chips. The HR-HPV infection rate was 81.9% with SCC samples showing the highest HR-HPV infection rate of 31%. CIN 3, CIS and CIN 2 showed infection rates of 25%, 16.4% and 9.5%, respectively. According to age with HR HPV infection rate, the 30~39 years-old group showed the highest infection rate by 92.3%. According to distribution with HR HPV genotyping, HPV 16 showed the highest infection rate by 42.3% whereas HPV 33 and HPV 58 showed infection rates of 11.7% and 10.8%, respectively. HPV 18 which is the second most common infected HPV genotype in the world showed 3.6%. Of the three most common oncogenic HR-HPV genotypes in CIN 2, we detected HPV 16, 35, 58; CIN 3 was HPV 16, 33, 58; CIS was HPV 16, 58, 33 (35/52); and SCC was HPV 16, 33, and 18 (31/52/58). Among the HPV 18, CIN 2, CIN 3, CIS and SCC showed 0.9%, 0.9%, 0% and 1.8%, respectively. The most often used preventive vaccines for cervical cancers use HPV 16 and HPV 18 as targets. However, results derived from this study suggest that a preventive vaccine against HPV 16 and HPV 18 would not be optimal for populations in this study.

Expression of Functional Pentameric Heat-Labile Enterotoxin B Subunit of Escherichia coli in Saccharomyces cerevisiae

  • Lim, Jung-Gu;Kim, Jung-Ae;Chung, Hea-Jong;Kim, Tae-Geum;Kim, Jung-Mi;Lee, Kyung-Ryul;Park, Seung-Moon;Yang, Moon-Sik;Kim, Dae-Hyuk
    • Journal of Microbiology and Biotechnology
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    • v.19 no.5
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    • pp.502-510
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    • 2009
  • Although the Escherichia coli heat-labile enterotoxin B subunit (LTB) has already been expressed in several different systems, including prokaryotic and eukaryotic organisms, studies regarding the synthesis of LTB into oligomeric structures of pentameric size in the budding yeast Saccharomyces cerevisiae have been limited. Therefore, this study used a functional signal peptide of the amylase 1A protein from rice to direct the yeast-expressed LTB towards the endoplasmci reticulum to oligomerize with the expected pentameric size. The expression and assembly of the recombinant LTB were confirmed in both the cell-free extract and culture media of the recombinant strain using a Western blot analysis. The binding of the LTB pentamers to intestinal epithelial cell membrane glycolipid receptors was further verified using a GM1-ganglioside enzyme-linked inmmunosorbent assay (GM1-ELISA). On the basis of the GM1-ELISA results, pentameric LTB proteins comprised approximately 0.5-2.0% of the total soluble proteins, and the maximum quantity of secreted LTB was estimated to be 3 mg/l after a 3-day cultivation period. Consequently, the synthesis of LTB monomers and their assembly into biologically active aligomers in a recombinant S. cerevisiae strain demonstrated the feasibility of using a GRAS microorganism-based adjuvant, as well as the development of carriers against mucosal disease.

Investigation of Nanofiber and Thermosensitive Scaffold for Intervertebral Disc through Organ Culture (기관배양을 통한 추간판 재생용 나노파이버 및 온도 감응성 지지체에 대한 검증)

  • Lee, Yong-Jae;Shin, Ji-Won;Shin, Ho-Jun;Kim, Chan-Hwan;Park, Ki-Dong;Bae, Jin-Woo;Seo, Hyoung-Yeon;Kim, Young-Jick;Shin, Jung-Woog
    • Journal of Biomedical Engineering Research
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    • v.28 no.4
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    • pp.512-519
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    • 2007
  • The purpose of this study is to investigate the potential of a novel tissue engineering approach to regenerate intervertebral disc. In this study, thermosensitive scaffold (chitosan-Pluronic hydrogel) and nanofiber were used to replace the nucleus pulposus (NP) and annulus fibrosus of a degenerated intervertebral disc, leading to an eventual regeneration of the disc using the minimally invasive surgical procedure and organ culture. In preliminary study, disc cells were seeded into the scaffolds and cellular responses were assessed by MTT assay and scanning electron microscopy (SEM). Based on these results, we could know that tissue engineered scaffolds might provide favorable environments for the regeneration of tissues. Organ culture was performed in fresh porcine spinal motion segments with endplates on both sides. These spinal motion segments were classified into three groups: control (Intact), injured NP (Defect), and inserting tissue engineered scaffolds (Insert). The specimens were cultivated for 7 days, subsequently structural stability, cell proliferation and morphological changes were evaluated by the relaxation time, quantity of DNA, GAG and histological examination. In these results, inserting group showed higher relaxation time, reduced decrement of DNA contents, and accumulated GAG amount. Consequently, the tissue engineered scaffolds used in this study seen to be a promising base scaffolds for regenerative intervertebral disc due to its capacity to absorb external dynamic loading and the possible ideal environment provided for disc cell growing.