• Title/Summary/Keyword: Micro-PCR

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Differential Expression of Nuclear Receptors in T Helper Cells

  • Hwang, Soo-Suk;Kim, Young-Uk;Lee, Won-Yong;Lee, Gap-Ryol
    • Journal of Microbiology and Biotechnology
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    • v.19 no.2
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    • pp.208-214
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    • 2009
  • Steroid hormones have long been known to have a profound influence on the immune system. Although the functions of the nuclear receptors in the development of T cells are fairly well studied, the differential expression of these receptors in T helper cells is poorly understood. Here, we investigated the differential expression of nuclear receptors and coregulators in Th1 and Th2 cells by genome-wide micro array analysis. The result showed that several nuclear receptors and coregulators are differentially expressed in these cells. The result was confirmed by RT-PCR. The result showed that $RXR{\alpha}$ is highly expressed in Th2 cells. Overexpression of $RXR{\alpha}$ in a Jurkat human T cell line induced IL4 but not IFN-${\gamma}$ gene expression, suggesting that $RXR{\alpha}$ plays a selective role in Th1 and Th2 differentiation. In summary, these results suggest that Th1/Th2 differentiation is influenced by differential regulation of nuclear receptors and coregulators.

DNA Microarray and Gene Ontology Enrichment Analysis Reveals That a Mutation in opsX Affects Virulence and Chemotaxis in Xanthomonas oryzae pv. oryzae

  • Kim, Hong-Il;Park, Young-Jin
    • The Plant Pathology Journal
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    • v.32 no.3
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    • pp.190-200
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    • 2016
  • Xanthomonas oryzae pv. oryzae (Xoo) causes bacterial leaf blight (BLB) in rice (Oryza sativa L.). In this study, we investigated the effect of a mutation in opsX (XOO1056), which encodes a saccharide biosynthesis regulatory protein, on the virulence and bacterial chemotaxis of Xoo. We performed DNA micro-array analysis, which showed that 63 of 2,678 genes, including genes related to bacterial motility (flagellar and chemotaxis proteins) were significantly downregulated ($<\;-2\;log_2$ fold changes) by the mutation in opsX. Indeed, motility assays showed that the mutant strain was nonmotile on semisolid agar swarm plates. In addition, a mutant strain (opsX::Tn5) showed decreased virulence against the susceptible rice cultivar, IR24. Quantitative real-time RT-PCR reaction was performed to confirm the expression levels of these genes, including those related to flagella and chemotaxis, in the opsX mutant. Our findings revealed that mutation of opsX affects both virulence and bacterial motility. These results will help to improve our understanding of Xoo and provide insight into Xoo-rice interactions.

Saliva Supernatant miR-21: a Novel Potential Biomarker for Esophageal Cancer Detection

  • Xie, Zi-Jun;Chen, Gang;Zhang, Xu-Chao;Li, Dong-Feng;Huang, Jian;Li, Zi-Jun
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.12
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    • pp.6145-6149
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    • 2012
  • Objective: To identify whether saliva supernatant miR-21 can serve as a novel potential biomarker in patients with esophageal cancer (EC). Methods: 32 patients with EC and 16 healthy controls were recruited in this study. Total RNA was extracted from saliva supernatant samples for measurement of miR-21 levels using RT-qPCR and relationships between miR-21 levels and clinical characteristics of EC patients were analyzed. Results: miR-21 was significantly higher in the EC than control groups. The sensitivity and specificity were 84.4% and 62.5% respectively. Supernatant miR-21 levels showed no significant correlation with cancer stage, differentiation and nodal metastasis. Conclusions: Saliva supernatant miR-21 may be a novel biomarker for EC.

Estradiol Valerate Exerts Neuroprotective Effects in Ischemic Rat Brain when Administered after Middle Cerebral Artery Occlusion

  • Yoo, Seong-Jin;Yu, Jeong-Min;Youm, Mi-Young;Kim, Do-Rim;Kim, Jee-Yun;Kang, Sung-Goo
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.111-111
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    • 2002
  • Stroke occurs when local thrombosis, embolic particle or the rupture of blood vessele interrupts the blood floe to the brain. $\beta$-estradiol 17-valerate has been reported to exert neuroprotective effects when administered before an ischemic insult. Recently, the pathophysiology of cerebral ischemia has been studied extensively in rat with various methods. In the present study, we investigates whether $\beta$-estrodiol 17-valerate can protect against brain injury. RNA sample were extracted from the hippocampus of female rat, reverse-transcription in the presence of [$\alpha$32p] dATP. Differential gene express-ion profiles were revealed (Bone morphogenetic protein type 1A receptor, Protein disulphide isomerase, Leukemia inhibitor factor receptor, cytochrome bc- 1 complex-x core P, thiol-specific antioxidant protein). RT-PCR was used to validate the relative expression pattern obtained by the cDNA array. The precise relationship between the early expression of recovery genes and stroke is a matter of luther investigation. This Study was supported by the Korea Science and Engineering Foundation(KOSEF) through the Biohealth Products Research Center(BPRC), Inje University, Korea.

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miR-181b as a Potential Molecular Target for Anticancer Therapy of Gastric Neoplasms

  • Guo, Jian-Xin;Tao, Qing-Song;Lou, Peng-Rong;Chen, Xiao-Chun;Chen, Jun;Yuan, Guang-Bo
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.5
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    • pp.2263-2267
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    • 2012
  • Objective: MicroRNAs (miRNAs) play important roles in carcinogenesis. The aim of the present study was to explore the effects of miR-181b on gastric cancer. Methods: The expression level of miR-181b was quantified by qRT-PCR. MTT, flow cytometry and matrigel invasion assays were used to test proliferation, apoptosis and invasion of miR-181b stable transfected gastric cancer cells. Results: miR-181b was aberrantly overexpressed in gastric cancer cells and primary gastric cancer tissues. Further experiments demonstrated inducible expression of miR-181b by Helicobacter pylori treatment. Cell proliferation, migration and invasion in the gastric cancer cells were significantly increased after miR-181b transfection and apoptotic cells were also increased. Furthermore, overexpression of miR-181b downregulated the protein level of tissue inhibitor of metalloproteinase 3 (TIMP3). Conclusion: The upregulation of miR-181b may play an important role in the progress of gastric cancer and miR-181b maybe a potential molecular target for anticancer therapeutics of gastric cancer.

Biodistribution and Genotoxicity of Transferrin-Conjugated Liposomes/DNA Complexes in Mice

  • Lee Sang Mi;Kim Jin-Seok;Oh Yu-Kyoung;Lee Yong-Bok;Sah Hongkee
    • Macromolecular Research
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    • v.13 no.3
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    • pp.218-222
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    • 2005
  • Transferrin-conjugated liposomes ($T_f$-liposomes) were made and formulated with pCMVluc DNA to form a lipoplex. Among the various formulations studied, the $T_f$-liposome: pCMVluc DNA complex at a ratio of 5: 1 (wt/wt) showed the highest transfection efficiency, which was twice that of $Lipofectin^{TM}$ on HeLa cells. The maxi-mum tolerated dose (MTD) of this lipoplex formulation from a single intravenous injection was over 10 mg/kg in healthy ICR mice. The RT-PCR results showed that the highest level of luciferase mRNA was detected in the lungs, followed by the liver, spleen, heart and kidneys, after an intravenous injection into mice. Two weeks after the injection, the levels of luciferase mRNA decreased gradually in the liver, spleen, heart, and kidney, but not in the lungs. The micro-array study showed that the cancer-related genes, including the bcl 6 gene, were highly up-regulated by the treatment with $T_f$-liposome/ pCMVluc DNA complex on HeLa cells, indicating that there were possible interactions between the host chromosomal DNA and the $T_f$-liposome within the cells. The results obtained from this study are expected to be useful for designing a safe and efficient gene delivery system using transferrin-conjugated liposomes.

Survey of Garlic Virus Disease and phylogenetic Characterization of Garlic Viruses of the Genus Allexivirus Isolated in Korea

  • Koo, Bong-Jin;Kang, Sang-Cu;Chang, Moo-Ung
    • The Plant Pathology Journal
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    • v.18 no.5
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    • pp.237-243
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    • 2002
  • A survey of virus infection in garlic plants cultivated in Korea was conducted for three years. Most virus-infected garlic plants (Allium sativum) showed typical symptoms on the leaves such as yellow mosaic, stripes, and distortion. Through immunosorbent electron micro-scopy and RT-PCR analysis, the complex mixtures of viruses including garlic viruses of the genus Allerivirus, gaylic strain of Leek yellow stripe virus of the genus Potyvirus, and Garlic latent virus of the genus Carlavirus were identified in the virus-infected garlic plants. Among these viruses, Allexivirus was the most frequently detect-ed in the regions surveyed. Using sets of differential primers for Allexivirus genomes, two members of the genus were amplified and sequenced from the purified viruses. The deduced amino acid sequences for the coat proteins and the nucleic acid binding proteins of two viruses showed high homologies to Garlic virus A (CarV-A) and Garlic virus D (GarV-D) of Allekivirus. This is the first report of GarV-A and GarV-D in Korea. This suggests that Allexivirus in gavlic plants in Korea was mixed and varied. Phylogenetic analyses showed that the genus Allexivirus was diversi(ied by the processes of accumulation and evolution of viruses in garlic plants due to the long period of repeated vegetative propagation.

Circulating miR-221 and miR-222 as Potential Biomarkers for Screening of Breast Cancer

  • Kim, Jungho;Oh, Sehee;Park, Sunyoung;Ahn, Sungwoo;Choi, Yeonim;Kim, Geehyuk;Kim, Seung Il;Lee, Hyeyoung
    • Biomedical Science Letters
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    • v.25 no.2
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    • pp.185-189
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    • 2019
  • Breast cancer is the second most common cancer in women with approximately 522,000 deaths annually worldwide. microRNAs have recently been studied as potential biomarkers that regulate gene expression and are involved in tumorigenesis. Here we evaluated circulating miR-221 and miR-222 as potential biomarkers for breast cancer by quantitative reverse transcription PCR using blood plasma of 30 healthy controls and 30 breast cancer patients. The TNM stage on circulating miR-221 and miR-222 was also investigated. Circulating miR-221 and miR-222 were significantly up-regulated in breast cancer patients compared to those in healthy controls (P < 0.0022 and P = 0.0058, respectively). Furthermore, the relative expression level of circulating miR-221 in patients with stage III breast cancer was higher than in those with stage I and II. Taken together, we have shown circulating miR-221 and miR-222 could be useful biomarkers for the screening of breast cancer patients.

Effects of Hansu-Daebowon (HDW) on RANKL-induced Osteoclast Differentiation and Bone Loss in Mammal Model (한수대보원이 포유동물인 생쥐 모델에서 골 손실 및 RANKL 유도 파골세포 분화에 미치는 영향)

  • Jang, Si-sung;Ryu, Hong-sun;Jeon, Chan-yong;Hwang, Gwi-seo
    • The Journal of Internal Korean Medicine
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    • v.40 no.1
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    • pp.58-69
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    • 2019
  • Objective: This study investigated the effects of Hansu-Daebowon (HDW) on bone resorption in vitro and bone loss in vivo. Methods: Osteoclast differentiation was measured by counting TRAP (+) MNC formed from RAW 264.7 in the presence of RANKL. Bone pit formation was determined in an artificial bone slice loaded with RANKL-stimulated osteoclasts. To elucidate the mechanisms of the inhibitory effects of HDW on bone resorption and osteoclast differentiation, osteoclastogenic genes (i.e. TRAP, MMP-9, NFATc1, c-Fos, and Cathepsin K) were measured using real time PCR. Furthermore, bone loss was observed using micro-CT in an LPS-treated mammal model. Results: HDW inhibited the bone pit formation in vitro and inhibited bone loss in vivo. Moreover, HDW decreased the number of TRAP (+) MNCs in the presence of RANKL, and HDW inhibited the expressions of cathepsin K, MMP-9, TRAP, NFATc1, and c-Fos in the osteoclasts. Conclusion: HDW exerts inhibitory effects on bone loss and bone resorption resulting from the inhibitions of osteoclast differentiation and osteoclastogenic gene expression.

Knockdown of Circ_0000144 Suppresses Cell Proliferation, Migration and Invasion in Gastric Cancer Via Sponging MiR-217

  • Ji, Fengcun;Lang, Chao;Gao, Pengfei;Sun, Huanle
    • Journal of Microbiology and Biotechnology
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    • v.31 no.6
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    • pp.784-793
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    • 2021
  • Previous studies have uncovered the role of circ_0000144 in various tumors. Here, we investigated the function and mechanism of circ_0000144 in gastric cancer (GC) progression. The expression of circ_0000144 in GC tissues and cells was detected through quantitative real-time polymerase chain reaction (qRT-PCR) method. Gain- and loss-of-function experiments including colony formation, wound healing and transwell assays were performed to examine the role of circ_0000144 in GC cells. Furthermore, western blot was conducted to determine the expressions of epithelial mesenchymal transition (EMT)-related proteins. The interaction between circ_0000144 and miR-217 was analyzed by bioinformatic analysis and luciferase reporter assays. The circ_0000144 expression was obviously upregulated in GC tissues and cells. Silencing of circ_0000144 inhibited cell proliferation, migration and invasion of GC cells, but ectopic expression of circ_0000144 showed the opposite results. Moreover, circ_0000144 sponged miR-217, and rescue assays revealed that silencing miR-217 expression reversed the inhibitory effect of circ_0000144 knockdown on the progress of GC. Our findings reveal that circ_0000144 inhibition suppresses GC cell proliferation, migration and invasion via absorbing miR-217, providing a new biomarker and potential therapeutic target for treatment of GC.