• The Korean Journal of Nuclear Medicine
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• v.36 no.1
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• pp.74-79
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• 2002

In addition to the well-established use of positron emission tomography (PET) in clinical oncology, novel roles for PET are rapidly emerging in the field of gene therapy. Methods for controlled gene delivery to living bodies, made available through advances in molecular biology, are currently being employed in animals for research purposes and in humans to treat diseases such as cancer. Although gene therapy is still in its early developmental stage, it is perceived that many serious illnesses could be treated successfully by the use of therapeutic gene delivery. A major challenge for the widespread use of human gene therapy is to achieve a controlled and effective delivery of foreign genes to target cells and subsequently, adequate levels of expression. As such, the availability of noninvasive imaging methods to accurately assess the location, duration, and level of transgene expression is critical for optimizing gene therapy strategies. Current endeavors to achieve this goal include methods that utilize magnetic resonance imaging, optical imaging, and nuclear imaging techniques. As for PET, reporter systems that utilize genes encoding enzymes that accumulate positron labeled substrates and those transcribing surface receptors that bind specific positron labeled ligands have been successfully developed. More recent advances in this area include improved reporter gene constructs and radiotracers, introduction of potential strategies to monitor endogenous gene expression, and human pilot studies evaluating the distribution and safety of reporter PET tracers. The remarkably rapid progress occurring in gene imaging technology indicates its importance and wide range of application. As such, gene imaging is likely to become a major and exciting new area for future application of PET technology.

• Nuclear Medicine and Molecular Imaging
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• v.42 no.5
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• pp.383-393
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• 2008

Purpose: Cancer specific killing can be achieved by therapeutic gene activated by cancer specific promotor. Expression of sodium iodide symporter (NIS) gene causes transportation and concentration of iodide into the cell, therefore radioiodine treatment after NIS gene transfer to cancer cell could be a form of radionuclide gene therapy. luciferase (Luc) gene transfected cancer cell can be monitored by in vivo optical imaging after D-luciferin injection. Aims of the study are to make vector with both therapeutic NIS gene driven by AFP promoter and reporter Luc gene driven by CMV promoter, to perform hepatocellular carcinoma specific radiodiodine gene therapy by the vector, and assessment of the therapy effect by optical imaging using luciferase expression. Materials and Methods: A Vector with AFP promoter driven NIS gene and CMV promoter driven Luc gene (AFP-NIS-CMV-Luc) was constructed. Liver cancer cell (HepG2, Huh-7) and non liver cancer cell (HCT-15) were transfected with the vector using liposome. Expression of the NIS gene at mRNA level was elucidated by RT-PCR. Radioiodide uptake, perchlorate blockade, and washout tests were performed and bioluminescence also measured by luminometer in these cells. In vitro clonogenic assay with 1-131 was performed. In vivo nuclear imaging was obtained with gamma camera after 1-131 intraperitoneal injection. Results: A Vector with AFP-NIS-CMV-Luc was constructed and successfully transfected into HepG2, Huh-7 and HCT-15 cells. HepG2 and Huh-7 cells with AFP-NIS-CMV-Luc gene showed higher iodide uptake than non transfected cells and the higher iodide uptake was totally blocked by addition of perchlorate. HCT-15 cell did not showed any change of iodide uptake by the gene transfection. Transfected cells had higher light output than control cells. In vitro clonogenic assay, transfected HepG2 and Huh-7 cells showed lower colony count than non transfected HepG2 and Huh-7 cells, but transfected HCT-15 cell did not showed any difference than non transfected HCT-15 cell. Number of Huh-7 cells with AFP-NIS-CMV-Luc gene transfection was positively correlated with radioidine accumulation and luciferase activity. In vivo nuclear imaging with 1-131 was successful in AFP-NIS-CMV-Luc gene transfected Huh-7 cell xenograft on nude mouse. Conclusion: A Vector with AFP promoter driven NIS and CMV promoter driven Luc gene was constructed. Transfection of the vector showed liver cancer cell specific enhancement of 1-131 cytotoxicity by AFP promoter, and the effect of the radioiodine therapy can be successfully assessed by non-invasive luminescence measurement.

• The Journal of Korean Medicine
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• v.27 no.1 s.65
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• pp.11-22
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• 2006

Objectives : In this study, the aim was to investigate the inhibitory effects of Bangpungtong-seoung-san on the obese gene and obese inhibitory about obese-mouse induced by high fat diet. Methods : In order to investigate the effects of Bangpuangtongseoung-san(BPTS) on the obese gene and obese inhibitory, C57BL/6 mice were induced by high fat diet. C57BL/6 mice were divided into three groups(normal, high fat diet with control, high fat diet with BPTS extract) and fed for 13weeks. Results : 1. The change of body weight and the final increase of body weight were decreased significantly. 2. The amount of the adipocyte in body weight was decreased significantly. 3. In primary adipocytes, ${\beta}3AR$ gene expression was increased significantly and leptin gene expression was decreased in $100{\mu}g/ml$ density significantly. 4. In primary adipocytes, the amount of TNF-${\alpha}$ was decreased in $100{\mu}g/ml$ density significantly and the amount of leptin was decreased but did not show significance. 5. In adipocytes tissue, the expression of leptin was decreased and the expression of ${\beta}3AR$ was increased. Conclusions : These results suggest that BPTS may inhibit the expression of the obese gene mad obese inhibitory about obese-mouse induced by high fat diet.

• Journal of Embryo Transfer
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• v.15 no.2
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• pp.157-165
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• 2000

The efficiency of transgenic livestock production could be improved by early screening of transgene-integration and sexing of embryos at preimplantational stages before trasferring them into recipients. We examined the effciency of multiplex PCR analysis for the simultaneous confirmation of the trasgene and sex during the preimplantational development of bovine embryos and the possibility of green fluorescent protein(GFP) gene as a non-invasive marker for the early screening of transgenic embryos. The GFP gene was microinjected into the male pronuclei of bovine zygotes produced in vitro. The injected zygotes were co-cultured in TCM-199 containing 10% FCS with boving oviductal epithelial cells in a 5% CO2 incubator. Seventeen(13.0%) out of 136 gene-injected bovine zygotes developed by multiplex PCR analysis and the expression of GFP was detected by observing green fluorescence in embryos under a fluorescent microscope. Eight(67%) of 12 embryos at 2-cell to blastocyst stage were positive in the PCR analysis, but only two(11.8%) of 17 blastocysts expressed the GFP gene. Their sex was determined as 7 female and 5 male embryos by the PCR analysis. The results indicate that the screening of GFP gene and sex in bovine embryos by PCR analysis and fluorescence detection could be a promisible method for the preselection of transgenic embryos.

• Molecular & Cellular Toxicology
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• v.5 no.4
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• pp.291-297
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• 2009

Kawasaki disease (KD) is believed to be infectious but etiology and the mechanism of development remain elusive. The aim of this study was to investigate the association between transmembrane channel-like 1 (TMC1) gene and KD. One hundred nine KD patients and 424 normal controls were enrolled. Of all KD patients, 34 developed coronary artery lesions (CALs). Eleven single nucleotide polymorphisms (SNPs) within TMC1 gene were selected and SNP genotyping was performed by the direct sequencing. Genotype frequencies were analyzed with the SNPAnalyzer, Helixtree, and SNPStats programs. In the present study, six SNPs (rs7851577, rs10781105, rs2589615, rs1663743, rs1373628, and rs1373626) were significantly associated with the risk of KD. In further haplotype analysis, one haplotype (CGGACCCT) showed a significant association between KD and control groups. These results suggest that TMC1 gene may be a susceptibility gene for KD in Korean population.

• Genomics & Informatics
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• v.4 no.3
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• pp.110-117
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• 2006

In microarray technology, many diverse experimental features can cause biases including RNA sources, microarray production or different platforms, diverse sample processing and various experiment protocols. These systematic effects cause a substantial obstacle in the analysis of microarray data. When such data sets derived from different experimental processes were used, the analysis result was almost inconsistent and it is not reliable. Therefore, one of the most pressing challenges in the microarray field is how to combine data that comes from two different groups. As the novel trial to integrate two data sets with batch effect, we simply applied standardization to microarray data before the significant gene selection. In the gene selection step, we used new defined measure that considers the distance between a gene and an ideal gene as well as the between-slide and within-slide variations. Also we discussed the association of biological functions and different expression patterns in selected discriminative gene set. As a result, we could confirm that batch effect was minimized by standardization and the selected genes from the standardized data included various expression pattems and the significant biological functions.

• Development and Reproduction
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• v.24 no.3
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• pp.197-205
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• 2020

Diabetes mellitus is a common heterogeneous metabolic disorder, characterized by deposition of extracellular matrix, oxidative stress, and vascular dysfunction, thereby leading to gradual loss of function in multiple organs. However, little attention has been paid to gene expression changes in the lung under hyperglycemic conditions. In this study, we found that diabetes inuced histological changes in the lung of streptozotocin-induced diabetic mice. Global gene expression profiling revealed a set of genes that are up- and down-regulated in the lung of diabetic mice. Among these, expression of Amigo2, Adrb2, and Zbtb16 were confirmed at the transcript level to correlate significantly with hyperglycemia in the lung. We further evaluated the effect of human umbilical cord-derived perivascular stem cells (PVCs) on these gene expression in the lung of diabetic mice. Our results show that administration of PVC-conditioned medium significantly suppressed Amig2, Adrb2, and Zbtb16 upregulation in these mice, suggesting that these genes may be useful indicators of lung injury during hyperglycemia. Furthermore, PVCs offer a promising alternative cell therapy for treating diabetic complications via regulation of gene expression.

• The Korean Journal of Physiology and Pharmacology
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• v.15 no.5
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• pp.251-258
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• 2011

Here we have investigated how lactosylceramide (LacCer) modulates gene expression of adhesion molecules in TNF-${\alpha}$ and IFN ${\gamma}$ (CM)-stimulated astrocytes. We have observed that stimulation of astrocytes with CM increased the gene expression of ICAM-1 and VCAM-1. D-Threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol (PDMP) and N-butyldeoxynojirimycin (NBDNJ), inhibitors of glucosylceramide synthase (GLS) and LacCer synthase (galactosyltransferase, GalT-2), inhibited the gene expression of ICAM-1 and VCAM-1 and activation of their gene promoter induced by CM, which were reversed by exogenously supplied LacCer. Silencing of GalT-2 gene using its antisense oligonucleotides also attenuated CM-induced ICAM-1 and VCAM-1 expression, which were reversed by LacCer. PDMP treatment and silencing of GalT-2 gene significantly reduced CM-induced luciferase activities in NF-${\kappa}B$, AP-1, GAS, and STAT-3 luciferase vectors-transfected cells. In addition, LacCer reversed the inhibition of NF-${\kappa}B$ and STAT-1 luciferase activities by PDMP. Taken together, our results suggest that LacCer may play a crucial role in the expression of ICAM-1 and VCAM-1 via modulating transcription factors, such as NF-${\kappa}B$, AP-1, STAT-1, and STAT-3 in CM-stimulated astrocytes.

• IMMUNE NETWORK
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• v.9 no.4
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• pp.138-146
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• 2009

Background: The MHC region of the chromosome contains a lot of genes involved in immune responses. Here we have investigated the mouse NG29/Cd320 gene in the centrometrically extended MHC region of chromosome 17. Methods: We cloned the NG29 gene by RT-PCR and confirmed the tissue distribution of its gene expression by northern blot hybridization. We generated the NG29 gene expression constructs and polyclonal antibody against the NG29 protein to perform the immunofluorescence, immunoprecipitation and flow cytometric analysis. Results: The murine NG29 gene and its human homologue, the CD320/8D6 gene, were similar in the gene structure and tissue expression patterns. We cloned the NG29 gene and confirmed its expression in plasma membrane and intracellular compartments by transfecting its expresssion constructs into HEK 293T cells. The immunoprecipitation studies with rabbit polyclonal antibody raised against the NG29-NusA fusion protein indicated that NG29 protein was a glycoprotein of about 45 kDa size. A flow cytometric analysis also showed the NG29 expression on the surface of Raw 264.7 macrophage cell line. Conclusion: These findings suggested that NG29 gene in mouse extended MHC class II region was the orthologue of human CD320 gene even though human CD320/8D6 gene was located in non-MHC region, chromosome 19p13.

• Tuberculosis and Respiratory Diseases
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• v.50 no.1
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• pp.67-75
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• 2001

Background : Two tumor suppressor genes, p53 and p16, which have different roles in controlling the cell cycle and inducing apoptosis, are frequently inactivated during carcinogenesis including lung cancer. Single tumor suppressor gene therapies using either with p53 or p16 have been studied extensively. However, there is a paucity of reports regarding a combined gene therapy using these two genes. Methods : The combined effect of p53 and p16 gene transfer by the adenoviral vector on the growth of lung cancer cell lines and its interactive mechanism was investigated. Results : An isobologram showed that the co-transduction of p53 and p16 exhibited a synergistic growth in hibitory effect on NCI H358 and an additive effect on NCI H23. Cell cycle analysis demonstrated the induction of a synergistic G1/S arrest by a combined p53 and p16 transfer. This synergistic interaction was again confirmed in a soft agar confirmed in a soft agar clonogenic assay. Conclusion : These observations suggest the potential of a p53 and p16 combination gene therapy as another potent strategy in cancer gene therapy.