• Title/Summary/Keyword: High-level expression

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Tissue factor expression is associated with recurrence in patients with non-metastatic colorectal cancer

  • Jung, Hee Jae;Kim, Hye Jin;Kaneko, Kensuke;Kazama, Yoshihiro;Kawai, Kazushige;Ishihara, Soichiro;Choi, Gyu-Seog
    • Korean Journal of Clinical Oncology
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    • v.14 no.2
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    • pp.128-134
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    • 2018
  • Purpose: Previous studies have addressed the role of the hypercoagulable state in the pathogenesis of cancer progression and metastasis. In this study, we investigated the association between coagulation factors, including tissue factor (TF) expression, platelet count, and fibrinogen level, and disease recurrence in patients with non-metastatic colorectal cancer. Methods: Patients who underwent curative resection for stage II or III colorectal cancer between 2000 and 2007 were included in this study. Data from a prospectively maintained database were retrospectively reviewed. TF expression was determined by immunohistochemistry using an anti-TF monoclonal antibody. The Kaplan-Meier method was used to estimate 5-year disease-free survival. Results: TF was highly expressed in 257 of 297 patients (86.5%). TF expression was not significantly associated with the platelet counts (P=0.180) or fibrinogen level (P=0.281). The 5-year disease-free survival rate was lower in patients with high TF expression than in patients with low TF expression (72.3% vs. 83.9%, P=0.074). In Cox hazard analysis, high TF expression was an independent risk factor for tumor recurrence (hazard ratio [HR] 2.446; 95% confidence interval [CI], 1.054-5.674; P=0.037). Undifferentiated histologic type (HR, 2.911; 95% CI, 1.308-6.481; P=0.009), venous invasion (HR, 2.784; 95% CI, 1.431-5.417; P=0.003), and lymph node metastasis (HR, 2.497; 95% CI, 1.499-4.158; P<0.001), were also significantly associated with disease recurrence. Conclusion: TF expression is associated with a recurrence in patients with non-metastatic colorectal cancer. However, further studies are required to clarify the underlying mechanisms relating TF expression with oncologic outcomes and its potential role as a therapeutic target.

Rapid Selection of Multiple Gene Integrant for the Production of Recombinant Hirudin in Hansenula polymorpha

  • Kim Hwa Young;Sohn Jung Hoon;Kim Chul Ho;Rao K. Jagannadha;Choi Eui Sung;Kim Myung Kuk;Rhee Sang Ki
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.1
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    • pp.1-6
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    • 2000
  • For the rapid selection of higher recombinant hirudin producing strain in a methylotrophic yeast Hansenula polymorpha, a multiple gene integration and dose-dependent selection vector, based on a telomere-associated ARS and a bacterial aminoglycoside 3-phosphotransferase (aph) gene, was adopted. Two hirudin expression cassettes (HV1 and HV2) were constructed using the MOX promoter of H. polymorpha and the mating factor $\alpha$ secretion signal of S. cerevisiae. Multiple integrants of a transforming vector containing hirudin expression cassettes were easily selected by using an antibiotic, G418. Hirudin expression level and integrated plasmid copy number of the tested transformants increased with increasing the concentration of G418 used for selection. The expression level of HV1 was consistently higher than that of HV2 under the similar conditions, suggesting that the gene context might be quite important for the high-level gene expression in H. polymorpha. The highest hirudin producing strain selected in this study produced over 96 mg/L of biologically active hirudin in a 500-mL flask and 165 mg/L in a 5-L fermentor.

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Lipopolysaccharide Synergizes with Interferon-${\gamma}$ to Induce Expression of Mig mRNA in Mouse Peritoneal Macrophages

  • Kim, Young-Ho;Kim, Hee-Sun
    • Journal of Microbiology and Biotechnology
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    • v.10 no.5
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    • pp.599-605
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    • 2000
  • Lipopolysaccharide (LPS) is responsible for the tissue injury that occurs following the invasion of multicelluar organisms by Gram-negative microbes. The effect of LPS on IFN-$\gamma$-induced chemokine Mig gene expression in mouse peritoneal macrophages was investigated. Very little Mig mRNA was detectable upon exposure to LPS without IFN-$\gamma$. Although LPS alone is only minimally effective, LPS plus IFN-$\gamma$ synergized to produce a high level of Mig mRNA in the peritoneal macrophages. This synergy was not dependent on a new protein synthesis, and was not controlled at the level of the gene transcription. Futhermore, LPS did not increase IFN-$\gamma$-induced Mig mRNA stability. Accordingly, it is suggested the LPS may synergize the expression of IFN-$\gamma$-induced Mig mRNA through a process that depends on a pretranscriptional level or concurrent Mig mRNA translation.

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Heat stress during summer reduced the ovarian aromatase expression of sows in Korea

  • Hwan-Deuk Kim;Sung-Ho Kim;Sang-Yup Lee;Tae-Gyun Kim;Seong-Eun Heo;Yong-Ryul Seo;Jae-Keun Cho;Min Jang;Sung-Ho Yun;Seung-Joon Kim;Won-Jae Lee
    • Korean Journal of Veterinary Service
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    • v.46 no.3
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    • pp.227-234
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    • 2023
  • It has been addressed that heat stress due to high atmospheric temperature during summer in Korea induces impaired release of reproductive hormones, followed by occurring abnormal ovarian cyclicity, lower pregnancy ratio, and reduced litter size. Therefore, the present study attempted to compare seasonal change (spring versus summer) of the ovarian aromatase expression, an enzyme for converting testosterone into estrogen. While serum estrogen level in summer group was significantly lower than that of spring group, testosterone was not different between groups. Consistent with estrogen level, the ovarian aromatase expression in summer at follicular phase was significantly lower than the counterpart of spring. The ovarian aromatase expression was positively related with serum estrogen level significantly (r=0.689; P=0.008) and strongly negative correlation was identified (r=-0.533; P=0.078) with atmospheric temperature. The ovarian aromatase expression was not detected in immature ovarian follicles but specifically localized in the granulosa cell layers in both seasons. However, the aromatase intensity in the granulosa cell layers was stronger in spring than summer. Because testosterone level was not different between groups, it could be concluded that the lower level of estrogen during summer might be derived by not lack of substrate but lower expression of ovarian aromatase by heat stress.

The Inhibitory Effects of Gamipalmul-tang(jiaweibawu-tāng) on the Obese-Mice Induced by High Fat Diet (가미팔물탕(加味八物湯)이 생쥐의 비만억제에 미치는 영향)

  • Kim, Hyo-Young;Heo, Dong-Seok
    • Journal of Korean Medicine Rehabilitation
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    • v.18 no.2
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    • pp.81-96
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    • 2008
  • Objectives : This study was performed to investigate inhibitory effects of Gamipalmul-tang($jiaweibawu-t{\bar{a}}ng$) on the hematological and histological changes of obese mice. Methods : C57BL/6 mice were divided into four groups (normal group, high fat diet with normal saline, high fat diet with reductil, high fat diet with Gamipalmul-tang($jiaweibawu-t{\bar{a}}ng$) and fed for 8 weeks. Body weight change, final increase of body weight, ALT, AST, creatinine, total cholesterol, LDL-cholesterol, HDL-cholesterol, triglyceride, glucose, cell viability by cytotoxicity, the expression of ${\beta}3AR$ in 3T3-L1 cell, the expression of leptin, ${\beta}3AR$ and serotonin in adipocyte tissue and size of adipocyte were observed in 8 weeks. Results : 1. In 3T3-L1 cell. the expression of ${\beta}3AR$ was increased significantly. 2. The final increase of body weight in obese-mouse were decreased significantly. 3. The level of AST, ALT were decreased significantly. 4. The level of LDL-cholesterol was decreased significantly and HDL-cholesterol was increased significantly. 5. The levels of triglyceride was decreased and leptin and glucose were decreased significantly. 6. In adipocyte tissue, the expression of ${\beta}3AR$ were increased significantly. 7. In adipocyte tissue, the expression of leptin and serotonin were decreased significantly. 8. The size of Adipocyte was decreased. Conclusions : On the basis of these results, we concluded that Gamipalmul-tang($jiaweibawu-t{\bar{a}}ng$) has inhibitory effects in rat.

Highly Efficient Gene Expression in Rabbit Synoviocytes Using EBV-Based Plasmid (가토 윤활막 세포에서 EBV-Based 플라스미드를 사용한 효율적인 유전자 발현)

  • Kim, Jin Young;Oh, Sang Taek;Youn, JeeHee;Lee, Suk Kyeong
    • IMMUNE NETWORK
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    • v.4 no.3
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    • pp.190-197
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    • 2004
  • Background: Rheumatoid arthritis (RA) is an autoimmune disorder characterized by chronic synovial inflammation which leads to joint destruction. Gene therapy of RA targets the players of inflammation or articular destruction. However, viral vectors have safety problems and side effects, while non-viral vectors suffer from inefficient gene transfer and fast loss of gene expression. To overcome the limits of non-vial vectors, an EBV-based plasmid which is known to exert prolonged high level gene expression can be used. Methods: pEBVGFP, pEBVIL-10, and pEBVvIL-10 were constructed by cloning GFP, IL-10, and vIL-10 genes into an EBV-based plasmid, respectively. The pGFP was used as a control plasmid. Each constructs were lipofected into HIG-82 rabbit synoviocytes. The expression of GFP was monitored by FACS and confocal microscopy. IL-10 and vIL-10 expressions were measured by ELISA. Results: GFP expression 2 days after transfection was achieved in 33.2% of cells. GFP-expressing cells transfected with pGFP decreased rapidly from 4 days after transfection and disappeared completely by 11 days. Cells transfected with pEBVGFP began to decrease slowly from 4 days. But GFP expression was detected for over 35 days. In addition, HIG-82 cells transfected with pEBVIL-10 ($44.6{\pm}1.5ng/ml$) or pEBVvIL-10 ($51.0{\pm}5.7ng/ml$) secreted these cytokines at high levels. High level cytokine production by hygromycin selection was maintained at least for up to 26 days after transfection. Conclusion: These results suggest that the EBV-based plasmid has a potential to improve non-viral gene transfer system and may be applicable to treat RA without the drawbacks of viral vectors.

Studies on the Effective Drug Delivery System Using Naked Plasmid DNA for the Erythropoietin Expression in vivo

  • Park, Yeong-Seop;Jeong, Dong-Geon;An, Jin-Ho;Choe, Cha-Yong;Ju, Hyeon
    • 한국생물공학회:학술대회논문집
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    • 2003.04a
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    • pp.582-586
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    • 2003
  • There has been interest in developing gene therapy based on naked plasmid DNA for treating serum protein deficiencies and human erythropoietin (hEPO) is one of the candidate for gene therapy being Investigated most enthusiastically. We constructed novel plasmid DNA vectors pVAC-hEPOI/II/III which contain one, two, three hEPO gene(s) respectively for producing high level expression and secretion of hEPO in vitro and in vivo. NIH3T3 and COS7 cells were transfected transiently with these vectors and increase in hEPO expression in medium reached 2-5 fold in comparison with pSecTagB-hEPO. Intra muscular administrations of pVAC-hEPOI/II/III vectors into mice resulted in high level secretion of hEPO in the serum and corresponding increases in hematocrit level. In conclusion the transduction efficiency of naked plasmid vectors is one of the critical factors of a gene delivery system and these novel plasmid vectors will contribute to various gene therapy based on naked plasmid DNA.

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Design and Cloning of the Gene for a Novel Insulin Analogue, $(B^{30}$-Homoserine) Human Insulin

  • Nam, Doo-H.;Ko, Jeong-Heon;Lee, Seung-Yup
    • Archives of Pharmacal Research
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    • v.16 no.4
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    • pp.271-275
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    • 1993
  • In order to prepare a novel human insulin analogue suhbstituted with homoserine at B$^{30}$ / position, (B$^{30}$ /-homoserine) human insulin, a synthetic gene was designed by linking directly a gene for B chain with that for A chain. This gene was constructed by enzymatic joining of 10 different synthetic oligonucleotides, and then inserted at the polylinker region of pUC19 plasmid. To achieve a high level of gene expression, the gene fusion technique region of pUC19 plasmid. To achieve a high level of gene expression, the gene fusion technique was employed using amino terminal regions of lacZ gene up to Clal or hpal, and either of them has been located under tac promoter. The chemical induction of these fused genes by isopropyl-.betha.-D-thiogalactopyranoside (IPTG) gave a satisfactory level of expression in Escherichia coli harboring the ocnstructed plasmids. It was observed that the fused gene product as a single chain insulin precusor was produced more than 30% of total cell protein of E. coli as a form of inclusion body.

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Expression of an Angiogenin Binding Peptide and Its Anti-Angiogenic Activity

  • Choi, Suk-Jung;Ahn, Mi-Won;Yoon, Kyoung-Bum;Park, Jong-Won
    • BMB Reports
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    • v.31 no.5
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    • pp.427-431
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    • 1998
  • In the previous report (Choi et al., 1997), the angiogenin binding peptides identified from a phage-peptide library were analyzed by using the fusion proteins composed of the Escherichia coli maltose binding protein and its corresponding peptides. However, it was difficult to obtain a sufficient amount of the fusion proteins required for further analysis because of the low expression level. We now report a high level expression of the fusion protein and analysis of its anti-angiogenin activity. The use of strong T7 promoter and removal of signal sequence allowed about a 20-fold increase in the expression efficiency of the fusion protein. We were able to obtain about 10 mg of purified fusion protein from one liter of culture. The purified fusion protein showed angiogenin-specific affinity and inhibited the binding of biotinylated actin to human angiogenin at $IC_{50}$ of 0.6 mM. Its anti-angiogenin activity was also revealed by the chorioallantoic membrane assay.

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The relationships between children's attachment to mother, anger and emotional.behavioral problems (아동의 모 애착 및 분노와 정서.행동문제간의 관계)

  • Chun, Eun-Chung;Lee, Jin-Suk
    • Journal of Families and Better Life
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    • v.28 no.6
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    • pp.157-167
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    • 2010
  • This study examined the relationships among children's attachment to the mother, trait anger, anger expression style, and emotional-behavioral problems in children. The subjects were 384 5th and 6th graders at elementary schools in J and G cities of Jeonbuk-Do. The instruments were Revised Inventory of Parent and Peer Attachment (IPPA-R), Stat-Trait Anger Expression Inventory - Korean version (STAXI-K), and Strength and Difficulty Questionnaire (SDQ). The major results are as follows; The level of children's attachment to the mother was negatively related to anger and emotional-behavioral problems, and the level of trait anger was positively related to emotional-behavioral problems. Children who have high anger expression and anger suppression tended to show more externalized behavior problems. Finally, the regression analysis showed that children's attachment to the mother and anger are revealed as significant factors in emotional-behavioral problems.