• Title/Summary/Keyword: Human Fibroblast

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Secretory Production of Recombinant Urokinase Kringle Domain in Pichia pastoris

  • Kim, Hyun-Kyung;Hong, Yong-Kil;Park, Hyo-Eun;Hong, Sung-Hee;Joe, Young-Ae
    • Journal of Microbiology and Biotechnology
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    • v.13 no.4
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    • pp.591-597
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    • 2003
  • Human urokinase kringle domain, sharing homology with angiostatin kringles, has been shown to be an inhibitor of angiogenesis, which can be used for the treatment of cancer, rheumatoid arthritis, psoriasis, and retinopathy. Here, the expression of the kringle domain of urokinase (UK1) as a secreted protein in high levels is reported. UK1 was expressed in the methylotrophic yeast Pichia pastoris GS115 by fusion of the cDNA spanning from Ser47 to Lys135 to the secretion signal sequence of ${\alpha}-factor$ prepro-peptide. In a flask culture, the secreted UK1 reached about 1 g/l level after 120h of methanol induction and was purified to homogeneity by ion-exchange chromatography. Amino-terminal sequencing of the purified UK1 revealed that it was cleaved at the Ste13 signal cleavage site. The molecular mass of UK1 was determined to be 10,297.01 Da. It was also confirmed that the purified UK1 inhibited endothelial cell proliferation stimulated by basic fibroblast growth factor, vascular endothelial growth factor, or epidermal growth factor, in a dose-dependent manner. These results suggest that a P. pastoris sytem can be employed to obtain large amounts of soluble and active UK1.

The Evaluation for Stabilization and Application of Microemulsion Containing Isoflavone (이소플라본을 함유한 마이크로에멀젼의 안정성 및 응용 평가)

  • Jeong, Noh-Hee;Moon, Young-Jin;Lee, Hyang-Woo;Nam, Ki-Dae
    • Journal of the Korean Applied Science and Technology
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    • v.19 no.3
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    • pp.167-173
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    • 2002
  • The isoflavone as a derivatives of flavone is colorless crystalline compounds. It acts on synthesis of fibronectine, collagen III, collagen I in human normal fibroblast by same biological activity to animal hormone. In this study, we tried to search and demonstrate system content rate of dermal translocation system for cosmetics using microemulsion containing isoflavone. The results of microemulsion stability test by centrifugation, storage in incubator and circulation chamber showed that separation of phase did not appear after 30 days. By the skin flexibility test, it has confirmed efficiency and effect as cosmetics materials. As the result, the microemulsion showed that skin flexibility factor improved up to 7.6%. We could confirm that O/W type microemulsion was stable system.

Functional Evaluation of the Rockbream (Oplegnathus fasciatus) Beta-actin Promoter as a Candidate Regulatory Element for DNA Vaccination

  • Kosuke, Zenke;Lee, Sang-Yoon;Kim, Ki-Hong;Nam, Yoon-Kwon
    • Fisheries and Aquatic Sciences
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    • v.12 no.2
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    • pp.98-103
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    • 2009
  • The potential utility of the rockbream (Oplegnathus fasciatus) $\beta$-actin 5'-upstream sequence as a regulatory element for DNA vaccination was evaluated based on in vitro and in vivo heterologous expression assays. In the in vitro transfection experiment, the efficacy of the rockbream $\beta$-actin promoter to drive the expression of a downstream lacZ gene was significantly higher (more than fourfold) than that of the human cytomegalovirus (hCMV) promoter in two fish cell lines (grunt Haemulon plumierii fin and bluegill Lepomis macrochirus fry cell lines). In contrast, the functional activity of the rockbream $\beta$-actin promoter was hardly detectable in a mammalian mouse embryonic fibroblast cell line. Rockbream skeletal muscles injected in vivo with a GFP reporter construct driven by the $\beta$-actin promoter displayed the significantly higher expression of a GFP protein (more than threefold) than did those injected with hCMV promoter driven construct. Data from this study suggest that the homologous rockbream $\beta$-actin promoter could be used as a potential regulator for DNA vaccination in this species.

Mechanism for Antioxidant Activity of Nardostachys chinensis root Extract

  • Heo, Jee-In;Kim, Jeong-Hyeon;Lee, Jeong-Min;Kim, Sung Chan;Park, Jae-Bong;Kim, Jaebong;Lee, Jae-Yong
    • Journal of Applied Biological Chemistry
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    • v.57 no.1
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    • pp.17-22
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    • 2014
  • Nardostachys chinensis (N. chinensis) has been used in traditional medicine as a sedative and analgesic. It has been reported that N. chinensis extract has an antioxidant activity. However, the mechanism has not been elucidated. In this study, we showed that FOXO3a was activated by N. chinensis extract. FOXO3a is a transcriptional factor that involved in cell cycle arrest, DNA repair, apoptosis, and detoxification of reactive oxygen spices (ROS). Protein level of FOXO3a was increased by N. chinensis extract whereas phospho-FOXO3a (Thr 32) was not changed. Promoter activities of target genes of FOXO3a such as MnSOD, p27, and GADD45 were increased by N. chinensis extract. Among target genes, protein level of MnSOD was increased by N. chinensis extract, and this leads to removal of ROS level in human embryonic fibroblast (HEF) cells. These results suggested that N. chinensis extract has an antioxidant activity by upregulation of MnSOD through FOXO3a activation.

The Flower Extract of Abelmoschus manihot (Linn.) Increases Cyclin D1 Expression and Activates Cell Proliferation

  • Park, Yea-In;Cha, Yeo-Eun;Jang, Minsu;Park, Rackhyun;Namkoong, Sim;Kwak, Jongbock;Jang, Ik-Soon;Park, Junsoo
    • Journal of Microbiology and Biotechnology
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    • v.30 no.7
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    • pp.1044-1050
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    • 2020
  • Abelmoschus manihot (Linn.) is a medicinal herbal plant that is commonly used to treat chronic kidney disease and hepatitis. However, its effect on cell proliferation has not been clearly revealed. In this report, we sought to determine the effect of the flower extract of A. manihot (FA) on cell proliferation. Based on our findings, FA increased the proliferation of human diploid fibroblast (HDF) and HEK293 cells. Through cell cycle analysis, FA was found to increase the number of HDF cells in the S phase and G2/M phase. FA also increased the expression of cyclin D1 and enhanced the migration of HDF cells. By administering FA to HDF cells with ≥30 passages, a decrease in the number of senescence-associated β galactosidase-positive cells was observed, thereby indicating that FA can ameliorate cellular senescence. Collectively, our findings indicate that FA increases cyclin D1 expression and regulates cell proliferation.

Clinical Application of Autologous Platelet-Rich Plasma (PRP) on Delayed Wound Healing of a Dog with Burns

  • Lee, Shinho;Cheong, Jongtae;Lee, Joo-Myoung
    • Journal of Veterinary Clinics
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    • v.35 no.5
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    • pp.229-232
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    • 2018
  • A 9-year-old intact female Poodle (weighing 3.6 kg) was presented for the treatment of a partial and full thickness burn that covering 45% of body including bilateral thigh and sacrolumbar region. Autologous platelet-rich plasma (PRP) stimulates angiogenesis, promoting vascular in-growth and fibroblast proliferation. On the unrecovered right thigh of the dog, autologous PRP was injected into the lesion after application of micro-needles. For macroscopic evaluation digital, photographs were taken from wounds at days 0, 3, 5, 13, 21, and 28. On the $3^{rd}$ day after PRP application, epithelialization was accelerated. Application of autologous PRP accelerated wound-healing rate and healing time in full thickness burns as well as secondary complications originating from unrecovered wounds. The delayed lesion was completely healed on the $28^{th}$ day by autologous PRP treatment. In human, PRP was increasingly used in the treatment of a variety of soft tissue in the management of chronic non-healing wounds. This study has shown that PRP treatment can be a valuable and effective aid on intractable wound healing in the dog with burns.

Study on the Effect of Injinwieryungtang(IJWRT) on hyperlipidemia (인진위영탕(茵蔯胃苓湯)이 고지혈(高脂血) 유발(誘發) 백서(白鼠)의 혈액학적(血液學的) 변화(變化)에 미치는 영향(影響))

  • Hwang, Woon-Kyu;Kim, Dong-Hee
    • Journal of Haehwa Medicine
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    • v.14 no.2
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    • pp.71-85
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    • 2005
  • As a long-term goal for the development of new prescriptions for hyperlipidemia, IJWRT was examined in the present study using a rat model in which the hyperlipidemia was induced. The major parameters related to lipid metabolism were investigated and the key findings are summarized below. IJWRT didn't show the cytotoxicity on Human Fibroblast Cells(hFCs). The body weight of hyperlipidemia-induced rats began to show lower body weight beginning one week after IJWRT treatment compared to non-treated control group animals. Cholesterol levels showed a significant decrease beginning three weeks after IJWRT treatment, compared to hyperlipidemia-induced control group. Total cholesterol levels in IJWRT treated animal group were significantly decreased compared to the hyperlipidemia-induced control group. Glucose levels in IJWRT treated animal group were significantly decreased compared to the hyperlipidemia-induced control group. Triglyceride levels in IJWRT treated animal group were significantly decreased compared to the hyperlipidemia-induced control group. SGOT levels in IJWRT treated animal group were insignificantly decreased compared to the hyperlipidemia-induced control group whereas there was significant change in SGPT levels. HDL-cholesterol levels were significantly increased in IJWRT treated animal group compared to the hyperlipidemia-induced control group. LDL-cholesterol levels were significantly decreased in IJWRT treated animal group compared to the hyperlipidemia-induced control group.

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The Preventative Effect of Gamibangpungtongsungsan (KBTS) on Hypertension (가미방풍통성산의 항고혈압 작용)

  • Ha, Yeo-Tae;Kim, Dong-Hee
    • Journal of Haehwa Medicine
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    • v.14 no.2
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    • pp.55-70
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    • 2005
  • In oriental medicine, Gamibangpungtongsungsan (KBTS) has been used as a therapeutic agent for the treatments of acute stage of cerebrovascular diseases and hypertension. In the present study, underlying mechanism on KBTS effects was investigated using spontaneously hypertensive rats (SHR) by determining related parameters such as blood pressure, heart-beat rates, and hormones and plasma constituents. The major finding are summarized as follows. 1. KBTS treatment at concentrations lower than $125\;{\mu}g/m{\ell}$ did not show any cytotoxicity on cultured human fibroblast cells. 2. KBTS treatment in SHR significantly decreased blood pressure and heart-beat rate compared with untreated control. 3. KBTS treatment in SHR decreased aldosterone levels in the blood compared with untreated control, but the difference was not statistically significant. 4. KBTS treatment in SHR significantly decreased dopamine, norepinephrine and epinephrine levels in the blood compared with untreated control. 5. KBTS treatment in SHR decreased plasma ion concentrations such as Na+, K+, Ca2+, Cl- compared with untreated control; decreases in Na+ and Cl- were statistically significant. 6. KBTS treatment in SHR significantly decreased TNF-$\alpha$, IL-6, and IL-10 levels in the blood compared with untreated control. Thus, the present data show evidence on anti-hypertension activity of KBTS in an experimental animal system, which can provide further insights into the development of anti-hypertension therapeutic agents.

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An Experimental Study on the Effect of Low Level Laser and Some Cytokines on Gene Expression of Human Gingival Fibroblasts (저출력레이저조사와 염증성 자극물질이 치은섬유아세포의 유전자 발현에 미치는 영향에 관한 실험적 연구)

  • Jung-Min Kim;Keum-Back Shin
    • Journal of Oral Medicine and Pain
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    • v.19 no.2
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    • pp.57-71
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    • 1994
  • Gingival fibroblasts were cultured and subjected to the test of Northern blot analysis for the demonstration of various mRNA expression in response to the low level laser treatment. For duplication of in vivo. Wound healing process, fibroblasts were pretreated with proinflammatory cytokine interleukin-1$\beta$(IL-1$\beta$) or mitogenic substance phorbol 12-myristate 13-acetate(PMA) prior to laser irradiation. The results were as follows : 1. By the laser irradiation, the gene expression of collagen type I was markedly increased I n gingival fibroblasts, especially in the case of PMA pretreatment. The gene expression of collagen type IV, however, was not only affected by laser irradiation but also by chemical cell stimulation. 2. Oncogene v-myc expression was affected by both laser irradiation and IL-1$\beta$ or PMA stimulation, But v-fos gene expression was not detected in any case of this experimental system. 3. Heat shock gene(Hsp 70)was expressed constiutively, but slightly increased by laser irradiation. 4. mRNA of fibroblast growth factor(FGF) was induced by both laser irradiation and IL-1$\beta$ or PMA treatment.

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Effects of Early Cell Damage from Repetitive Intermittent Fever Exposure in Alopecia Progression and Evaluation of New Candidate Drugs: Ibuprofen, Menthol, and Cetirizine (간헐적 발열 반응에 의한 세포 손상과 이와 관련된 탈모 치료를 위한 신 후보물질 연구)

  • Lim, Sung Cil;Moon, Hong Seop
    • Korean Journal of Clinical Pharmacy
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    • v.26 no.3
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    • pp.187-194
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    • 2016
  • Background: Alopecia areata (AA) is a very disturbing and expensive disorder in which the exact etiology is not known and it is yet to be treated completely well. Most alopecia patients exhibit some inflammation in the hair follicles regardless of the causes. The clinical symptoms of alopecia present very diversely while the prime symptom is local intermittent fever which are related to inflamed cells. Methods: This study aimed to evaluate how repetitive intermittent fever can damage the normal human dermal fibroblast (NHDF) cells and investigated the cytotoxic and proliferative effects after application of new candidate drugs (ibuprofen, menthol, cetirizine) for alopecia in comparison to minoxidil. Results: This study demonstrated that ibuprofen, menthol, or/and cetirizine can prevent or slow down the damage of NHDF cells from intermittent fever in early alopecia. Aggressive preventative intervention with those drugs before complete destruction of hair follicle by excessive repetitive fever, is a very important step for alopecia therapy and these drugs are recommended as candidate drugs for alopecia in the future. Conclusion: Aggressive preventative intervention with drugs before complete destruction of hair follicles (NHDF cells) by excessive repetitive fever is a very important step for alopecia therapy or progression.