• Title/Summary/Keyword: Gene Delivery

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Roles of TLR-4 and NF-κB in Interleukin-6 Expression Induced by Heat Shock Protein 90 in Vascular Smooth Muscle Cells (혈관평활근세포에서 HSP90에 의한 IL-6 발현에 TLR-4와 NF-κB의 작용)

  • Rhim, Byung-Yong;Kim, Kang-Seong;Kim, Koan-Hoi
    • Journal of Life Science
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    • v.18 no.12
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    • pp.1637-1643
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    • 2008
  • This study has investigated whether extracellular HSP90 predisposes vascular smooth muscle cells (VSMCs) to pro-inflammatory phenotype. Exposure of rat aortic smooth muscle cells to HSP90 not only enhanced IL-6 release but also profoundly induced IL-6 transcript via promoter activation. HSP90-induced IL-6 promoter activation was suppressed by dominant-negative forms of Toll-like receptor (TLR)-4 and myeloid differentiation factor 88 (MyD88), but not by dominant-negative-forms of TLR-3 and TIR-domain-containing adapter-inducing interferon-${\beta}$ (TRIF). Curcumin, which inhibits dimerization of TLR-4, also attenuated the IL-6 induction by HSP90. Mutation at the NF-${\kappa}B$- or C/EBP-binding site in the IL-6 promoter region suppressed the promoter activation in response to HSP90. The gene delivery of $I{\kappa}B$ using recombinant adenoviruses and treatment with resveratrol, which inhibit NF-${\kappa}B$ activity, attenuated the HSP90-induced IL-6 release from VSMCs. The present study proposes that extracellular HSP90 would contribute to inflammatory reaction in the stressed vasculature by inducing IL-6 in VSMCs, and that TLR-4 and NF-${\kappa}B$ would play active roles in the process.

Effect of cultivar and ascorbic acid on in vitro shoot regeneration and development of bombardment-mediated plastid transformation of tomato (Lycopersicon esculentum) (토마토 재분화 효율 향상 및 엽록체 형질전환 조건)

  • Roh, Kyung-Hee;Lee, Ki-Jong;Park, Jong-Sug;Kim, Jong-Bum;Lee, Seung-Bum;Suh, Seok-Cheol
    • Journal of Plant Biotechnology
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    • v.37 no.1
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    • pp.77-83
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    • 2010
  • Eighteen cultivars of tomato were tested for their regeneration response. Lycopersicon esculentum cv. 2001-58 showed a very high frequency of regeneration (93%). We evaluated the effect of two compounds with known antioxidant activity (ascorbic acid and cystein). The use of ascorbic acid ($200\;-\;300\;{\mu}M/L$) has a positive effect on shoot regeneration. To develope a system for plastid transformation in tomato via homologous recombination, we constructed the tomato plastid expression vector (pKRT22-AG) harboring 2.2 kb flanking sequences cloned from intact plastid genome and gfp gene. To investigate the factors affecting the delivery system of the pKRT22-AG into chloroplast using bombardment, We assessed the optimal DNA concentration, gold particle volume and target distance. Expression of the GFP protein was observed within chloroplast on protoplast of cotyledon explant by confocal laser scanning microscopy, which indicates that the protocol developed in this study be useful for the production of plastid transgenic plants in tomato.

Oncolytic Viruses - A New Era for Cancer Therapy (종양 용해성 바이러스-암 치료에서의 새 시대)

  • Ngabire, Daniel;Niyonizigiye, Irvine;Kang, Min-jae;Kim, Gun-Do
    • Journal of Life Science
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    • v.29 no.7
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    • pp.824-835
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    • 2019
  • In recent decades, oncolytic viruses (OVs) have extensively been investigated as a potential cancer drug. Oncolytic viruses have primarily the unique advantage in the fact that they can only infect and destroy cancer cells. Secondary, oncolytic viruses induce the activation of specific adaptive immunity which targets tumor-associated antigens that were hidden during the initial cancer progression. In 2015, one genetically modified oncolytic virus, talimogene laherparepvec (T-VEC), was approved by the American Food and Drug Administration (FDA) for the treatment of melanoma. Currently, various oncolytic viruses are being investigated in clinical trials as monotherapy or in combination with preexistent cancer therapies like immunotherapy, radiotherapy or chemotherapy. The efficacy of oncolytic virotherapy relies on the balance between the induced anti-tumor immunity and the anti-viral response. Despite the revolutionary outcome, the development of oncolytic viruses for the treatment of cancer faces a number of obstacles such as delivery method, neutralizing antibodies and induction of antiviral immunity due to the complexity, variability and reactivity of tumors. Intratumoral administration has been successful reducing considerably solid tumors with no notable side effects unfortunately some tumors are not accessible (brain) and require a systemic administration of the oncolytic viruses. In order to overcome these hurdles, various strategies to enhance the efficacy of oncolytic viruses have been developed which include the insertion of transgenes or combination with immune-modulatory substances.

Association of Pre- and Perinatal Risk Factors With Tourette Syndrome or Chronic Tic Disorders in a Korean School-Age Population

  • Wooseok Choi;Soon-beom Hong;Johanna Inhynag Kim;Jung Lee;Soomin Jang;Yebin D Ahn;You Bin Lim;Sumin Kim;Mee Rim Oh;Bung-Nyun Kim
    • Journal of the Korean Academy of Child and Adolescent Psychiatry
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    • v.34 no.1
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    • pp.37-44
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    • 2023
  • Objectives: Tic disorders are highly heritable; however, growing evidence suggests that environmental factors play a significant role in their pathogenesis. Studies on these factors have been inconsistent, with conflicting results. Therefore, this study aimed to examine the associations of pre- and perinatal exposure to Tourette syndrome (TS) or chronic tic disorders (CTD) in Korean school-aged children. Methods: This case-control study used data from a large prospective cohort study. The primary outcome was TS/CTD diagnosis according to the Diagnostic and Statistical Manual of Mental Disorders, 5th edition (DSM-5) criteria and Kiddie-Schedule for Affective Disorders and Schizophrenia-Present and Lifetime Version-Korean Version. Demographic, pre-, and perinatal information was obtained from the maternal questionnaires. Data between the TS/CTD and control groups were compared using the chi-squared or Student's t-test, as appropriate. Two-step logistic regression analyses were used to test the association between TS/CTD and pre- and perinatal risk factors. Results: We included of 223 children (78 with TS/CTD and 145 controls). Significant differences in the demographic data between the two groups were observed. The male sex ratio, mean parental age, parental final education level, and family history of tics were included as confounders. In the final adjusted multivariable model, TS/CTD was significantly associated with antiemetic exposure during pregnancy (odds ratio [OR]=16.61, 95% confidence interval [CI] 1.49-185.22, p=0.02) and medically assisted reproduction (OR=7.89, 95% CI 2.28-27.28, p=0.01). Conclusion: Antiemetic exposure and medically assisted reproduction are significantly associated with the risk of TS/CTD. These results should be replicated in future prospective and gene-by-environment studies.

Adsorption of Glycerol on Hydroxyapatite Enhanced Colloidal Stability in Phosphate Buffered Saline Solution (글리세롤 흡착으로 인산완충식염수에서 콜로이드 안정성이 향상된 수산화인회석 합성)

  • Jaun An;Hyebin Choi;Keunyoung Lee;Ki-Young Kwon
    • Applied Chemistry for Engineering
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    • v.34 no.6
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    • pp.670-673
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    • 2023
  • The biocompatibility of hydroxyapatite (HAP) has led to its application in various fields. To accomplish practical biological applications, such as drug/gene delivery, the colloidal stability of HAP in phosphate-buffered saline (PBS) is particularly important. In this study, we prepared a glycerol incorporated-HAP (Gly-HAP) by heating HAP in a glycerol environment at 200 ℃. To compare morphology and colloidal stability, HAP prepared at room temperature (RT-HAP) was thermally treated in water at 200 ℃ (H2O-HAP). The heat treatment of HAP in both water and glycerol solutions results in an increase in the crystallinity of HAPs. Due to the low solubility of HAP in glycerol and the adsorption of glycerol on the HAP surface, crystal growth is limited. However, the heat-treated HAP under water increased in size by approximately four times compared to the initial crystallites. Compared to RT-HAP and H2O-HAP, Gly-HAP shows improved colloidal stability in PBS, which originates from the adsorption of glycerol on the HAP surface that inhibits the agglomeration of individual HAP precipitates.

A Study on the Labeling Efficiency and Cytotoxicity of Hepatocyte-targeting Galactosylated Chitosan Compounds (간세포 지향성 Galactosylated Chitosan 화합물의 표지 수율 향상 및 세포 독성에 대한 연구)

  • Kim, Dae-Weung;Jeong, Hwan-Jeong;Kim, Eun-Mi;Kim, Se-Lim;Kang, Yun-Hee;Kim, Min-Woo;Kim, Chang-Guhn;Sohn, Myung-Hee
    • The Korean Journal of Nuclear Medicine
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    • v.39 no.5
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    • pp.278-283
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    • 2005
  • Purpose: In prior study, we synthesized $^{99m}Tc$-galactosylated chitosan (GC) and performed in vivo biodistribution study, showed specific targeting to hepatocyte. The aim of this study is to evaluate the labeling efficiency and cytotoxicity of modified galactosylated chitosan compounds, galactosyl methylated chitosan (GMC) and HYNIC-galactosylated chitosan (GCH). Materials and Methods: GC, GMC and GCH were synthesized and radiolabeled with $^{99m}Tc$. Then, they were incubated for 6 hours at room temperature and human serum at $37^{\circ}C$. Labeling efficiencies were determined at 15, 30 m, 1, 2, 3 and 6 h after radiolabeling. To evaluate cytotoxicity, MTT assay was performed in HeLa and HepG2 cells. Results: In comparison with them of $^{99m}Tc$-GC labeling efficiencies of $^{99m}Tc$-GMC were significantly improved (100, 97 and 89%) in acetone and 96.3, 95.8 and 75.6% in saline at 15 m, 1 and 6 h, respectively). Moreover, $^{99m}Tc$-GCH showed more improved labeling efficiencies (>95% in acetone and human serum and >90% in saline at 6 h). In MTT assay, cytotoxicity was very low and not different from that of controls. Conclusion: These results represent that these compounds are radiochemically compatible radiopharmaceuticals, can be used in hepatocyte specific imaging study and in vivo gene or drug delivery monitoring.

Up-regulation of Pluripotency-related Genes in Human Amniotic Fluid-derived Stem Cells by ESRRB Conjugated with Cell-Penetrating Peptide (인간 양수 유래 줄기세포에서 세포투과단백질을 이용한 ESRRB 단백질의 직접도입에 의한 전분화능 관련 유전자의 발현 조절)

  • Jo, Jung-Hyun;Lee, Yu-Sun;Oh, Mi-Hee;Ko, Jung-Jae;Cheon, Yong-Pil;Lee, Dong-Ryul
    • Development and Reproduction
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    • v.14 no.4
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    • pp.243-251
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    • 2010
  • ESRRB (Estrogen related receptor $\beta$) is an orphan receptor, and have a role on maintaining the undifferentiated state and self-renewal of pluripotent stem cell as a transcription factor which regulates the expression of OCT4 and NANOG genes. Also, Feng et al. (2009) reported that Esrrb, Oct4 and Sox2 could induce pluripotent stem cell from somatic cells. The aim of the present study was to develop the direct delivery system of human ESRRB protein into human amniotic fluid-derived stem cells (AFSCs) and to analyze the effect of ESRRB on the regulation of pluripotency-related genes. Human ESRRB has three isoforms arisen by alternative splicing. We cloned short-form ESRRB and made a fusion protein of ESRRB and R7 for an efficient protein transfer to cell. R7 as cell-penetrating peptide(CPP) can help to transfer ESRRB into cells. R7-ESRRB-His6 protein was observed in the cytoplasm and nuclei within 5 hours after treatment. Also, we could observe R7-ESRRB-His6 protein only in the nuclei within 24 hours. Realtime PCR showed that ESRRB increased expression of OCT4 and NANOG as well as SOX2 gene. Therefore, we demonstrated that R7-ESRRB-His6 proteins were efficiently transferred into the nuclei of AFSCs and work well as a possible transcription factor.

A STUDY ON 4 TYPE CONSTITUTION AND SIFE CHARACTER OF OBESE PATIENTS (비만인의 생활특성과 사상체질에 관한 연구)

  • KIM, DAL RAE
    • Journal of Sasang Constitutional Medicine
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    • v.9 no.1
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    • pp.303-313
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    • 1997
  • Disease depends on the three factors, agent, host and environment. According to history of disease, by early 1900s the case of deaths is infectious disease, in late 1900s care of infectious diseases and tremendous scale of chronic disease, i.e., heart disease, diabetes, cancers and etc, makes care of chronic diseases be a most important theme. Now, life-style of diet is being westernized and in high industry-oriented society, obesity makes attack fate remarkably increase and life-expectancy become short, so that it causes severe problem of health. Chronic disease, such as obesity, is not affected by specific agent, but depends of interaction between host and environmental factors. There is the theory of constitutional medicine in Korean Medicine. According to it, all the people have constitutional specificity and disease. Because obesity is a kind of disease, there is the corresponding constituent being apt to be fat. Oriental Medicine utilizes herb-medication, acupuncture, and massage-therapy in treating obesity. Therefore study on relationship between constituent and obesity for OPD patients of Sangji-Oriental Medicine Hospital is carried out. The results are summarized as followings. 1. 70.2% of obesity patients are Taeumin(太陰人), 26.9% of those are Soyangin(少陽人), 2.9% of thoese are Soeumin(少陰人). 2. Most cases, high value of Free Fat Acid and Triglyceride not that of Total Cholesterol and Low Density Lipoprptein is meaningful in obesity patient blood. The corelationship between lipid test and Constitution is meaningful in Triglyceride and Free Fatty Acid. 3. Obesity is not related with gene. 4. Obesity is not related with Boyak(Herb-Med : 補藥). 5. Obesity mostly happens after delivery, contraception and operation. 6. Obese Patients are apt to eat between meals, especially food of wheat flour such as a snack. 7. The aim of treating obesity is not persuit of beauty but of keeping healthy. 8. 2.2Kg of body weight is lost after 4 week-treatment. 9. Common cause of obesity is overeating of carbohydrate and lipid than meat.

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Genetic Polymorphism in Corticotropin-releasing Hormone Receptor Type-1 in Preeclamptic Korean Women

  • Lim, Ji-Hyae;Kim, Shin-Young;Park, So-Yeon;Kim, Do-Jin;Kim, Mi-Jin;Ahn, Hyun-Kyong;Han, Jung-Yeol;Kim, Moon-Young;Park, Hyun-Young;Lee, Kwang-Soo;Kim, Young-Ju;Ryu, Hyun-Mee
    • Journal of Genetic Medicine
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    • v.8 no.2
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    • pp.113-118
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    • 2011
  • Purpose: Placental corticotropin-releasing hormone receptor type 1 (CRHR1) expression is reduced in pregnancies with abnormal placental function such as preeclampsia (PE), and the levels and/or function of CRHR1 are genetically influenced. The aim of this study was to investigate the association between the c.33+8199C>T polymorphism in the CRHR1 gene and PE in a Korean population. Materials and Methods: Using a case-control design, the association between the CRHR1 polymorphism and the risk of PE was investigated in 203 individuals with PE and 211 normotensive controls. Genotypes were determined using a SNapShot kit and an ABI Prism 3100 Genetic analyzer. Results: Genotypes and allele frequencies for the CRHR1 polymorphism did not differ between PE and normotensive pregnancies. The variant T allele was more frequent than the ancestral C allele in both of the groups and was more frequent in the controls than in the cases. In risk analysis for PE, there was not an increased risk of preeclampsia in subjects who were concomitant homozygous rare allele genotypes (CC) (OR, 0.3; P=0.15) or heterozygous rare allele genotypes (TC) (OR, 0.8; P=0.29). There were no differences in the complications of PE such as severity or preterm delivery in patients with the CRHR1 polymorphism. Conclusion: Our findings indicate that the CRHR1 polymorphism was not associated with PE in the present Korean study group.

Expression of Cell Cycle Related Genes in HL60 Cells Undergoing Apoptosis by X-irradiation (HL60 세포주에서 방사선 조사에 의한 Apoptosis와 세포 주기 관련 유전자의 발현 변화)

  • Kim, Jin-Hee;Park, In-Kyu
    • Radiation Oncology Journal
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    • v.16 no.4
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    • pp.377-388
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    • 1998
  • Purpose : To evaluate changes in expression of cell cycle related genes during apoptosis induced in HL60 cells by X-irradiation to understand molecular biologic aspects in mechanism of radiation therapy. Material and Methods : HL-60 cell line (promyelocytic leukemia cell line) was grown in culture media and irradiated with 8 Gr by linear accelerator (6 MV X-ray). At various times after irradiation, ranging from 3 to 48 hours were analyzed apoptotic DNA fragmentation assay for apoptosis and by western blot analysis and semi-quantitative RT-PCR for expression of cell cycle related genes (cyclin A, cyclin B, cyclin C, cyclin Dl, cyclin E, cdc2, CDK2, CDK4, $p16^{INK4a}$, $p21^{WAF1}$, $p27^{KIP1}$, E2F, PCNA and Rb). Results : X-irradiation (8 Gy) induced apoptosis in HL-60 cell line. Cycline A protein increased after reaching its peak 48 h after radiation delivery and cyclin E, E2F, CDK2 and RB protein increased then decreased after radiation. Radiation induced up-regulation of the expression of E2F is due to mostly increase of Phosphorylated retinoblastoma proteins (ppRb). Cyclin Dl, PCNA, CDC2, CDK4 and $p16^{INK4a}$ protein underwent no significant change at any times after irradiation. There was not detected $p21^{WAF1}$ and $p27^{KIP1}$ protein. Cyclin A, B, C mRNA decreased immediately after radiation and then increased at 12 h after radiation. Cyclin Dl mRNA increased immediately and then decreased at 48 h after radiation. After radiation, cyclin E mRNA decreased with the lapse of time. CDK2 mRNA decreased at 3h and increased at eh after radiation. CDK4 mRNA rapidly increased at 6 to 12 h after radiation. There was no change of expression of $p16^{INK4a}$ and not detected in expressin of $p21^{WAF1}$ and $p27^{KIP1}$ mRNA. Conclusion : We suggest that entry into S phase may contribute to apoptosis of HL60 cells induced by irradiation. Increase of ppRb and decrease of pRb protein are related with radiation induced auoptosis of HL60 cells and tosis of HL60 cells induced by irradiation. Increase of ppRb and decrease of PRb protein are related with radiation induced apoptosis of HL60 cells and this may be associated with induction of E2F and cyclinE/CDK2. These results support that $p21^{WAF1}$ and $p27^{KIP1}$ are not related with radiation induced-apoptosis.

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