• Journal of Yeungnam Medical Science
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• v.24 no.2
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• pp.262-274
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• 2007

Purpose : Bone morphogenetic proteins (BMPs) play an important role in the formation of cartilage and bone, as well as regulating the growth of chondroblasts and osteoblasts. In this study, we investigated whether recombinant human BMP adenoviruses are available for ex vivo gene therapy, using human fibroblasts and human bone marrow stromal cells in an animal spinal fusion model. Materials and Methods : Human fibroblasts and human bone marrow stromal cells were transduced with recombinant BMP-2 adenovirus (AdBMP-2) or recombinant BMP-7 adenovirus (AdBMP-7), referred to as AdBMP-7/BMSC, AdBMP-2/BMSC, AdBMP-7/HuFb, and AdBMP-2/HuFb. We showed that each cell secreted active BMPs by alkaline phosphatase staining. Since AdBMP-2 or AdBMP-7 tranducing cells were injected into the paravertebral muscle of athymic nude mice, at 4 weeks and 7 weeks, we confirmed that new bone formation occurred by induction of spinal fusion on radiographs and histochemical staining. Results : In the region where the AdBMP-7/BMSC was injected, new bone formation was observed in all cases and spinal fusion was induced in two of these. AdBMP-2/BMSC induced bone formation and spinal fusion occurred among one of five. However, in the region where AdBMP/HuFb was injected, neither bone formation nor spinal fusion was observed. Conclusion : The osteoinductivity of AdBMP-7 was superior to that of AdBMP-2. In addition, the human bone marrow stromal cells were more efficient than the human fibroblasts for bone formation and spinal fusion. Therefore, the results of this study suggest that AdBMP-7/BMSC would be the most useful approach to ex vivo gene therapy for an animal spinal fusion model.

• Tuberculosis and Respiratory Diseases
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• v.63 no.1
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• pp.42-51
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• 2007

Background: TRAIL is a cytokine that selectively induces apoptosis in various cancer cell lines. Gefitinib is new targeted drug applied in lung cancer that selectively inhibits EGFR tyrosine kinase. However, lung cancers have shown an initial or acquired resistance to these drugs. This study examined the effect of IGF-1R and its blockade on enhancing the sensitivity of lung cancer cell lines to TRAIL and gefitinib. Methods: Two lung cancer cell lines were used in this study. NCI H460 is very sensitive to TRAIL and gefitinib. On the other hand, A549 shows moderate resistance to TRAIL and gefitinib. The IGF-1R blockade was performed using adenoviruses expressing the dominant negative IGF-1R and shRNA to IGF-1R and AG1024 (IGF-1R tyrosine kinase inhibitor). Results: The adenovirus expressing dominant negative IGF-1R(950st) induced the increased expression of defective IGF-1R on the lung cancer cell surface, and the adenovirus-shIGF-1R effectively decreased the level of IGF-1R expression on cell surface. The genetic blockade of IGF-1R by the adenovirus-dnIGF-1R and AG1024 increased the sensitivity of A549 cells to TRAIL. The reduction of IGF-1R by transduction with ad-shIGF-1R also increased the sensitivity of the A549 cells to gefitinib. Conclusion: The blockade of IGF-1R through various mechanisms increased the sensitivity of the lung cancer cell line that was resistant to TRAIL and gefitinib. However, further studies using other cell lines showing acquired resistance as well as in vivo animal experiments will be needed.

• Pediatric Infection and Vaccine
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• v.10 no.1
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• pp.87-94
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• 2003

Purpose : Respiratory viruses are one of the most infectious agent in human. Acute lower respiratory tract infection(ALRTIs) is associated with significant morbidity and mortality in children. This study is performed to investigate the etiologic organism, age and sex distribution, clinical manifestations and seasonal occurrence of ALRTIs in children. Methods : Viral agent was evaluated with nasopharyngeal aspirates, rhinorrhea and saliva collected from 568 patients. We confirmed viral agents in 54 patients who were younger than 15 year old. They had visited Maryknoll Hospital, Busan in Korea from January, 2002 to December, 2002 for ALRTIs. Results : The viral pathogens identified were Influenza A virus(59.3%), Enterovirus(33.3%), Adenovirus(5.6%), and Influenza B virus(1.9%). Parainfluenza virus and Respiratory syncytial virus were not detected. The occurrence of acute lower respiratory infections was high between 3 & 6 years old. The clinical patterns include pneumonia(51.9%), bronchitis(31.5%), croup(9.3%), bronchiolitis(7.4%). The respiratory viral agents had their characteristic seasonal patterns. Conclusion : Influenza A virus was the most common cause of acute lower respiratory tract infections in Busan area during the 2002. ALRTIs had high occurrence between 3 to 6 years old. And the most common clinical patterns were pneumonia and bronchitis.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.27 no.2
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• pp.103-109
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• 2005

In spite of the ongoing advances, standard therapies for oral cancer still has some limitations in efficacy and in ability to prolong survival rate of advanced disease and result in significant functional defect and severe cosmetic deformity. Currently gene therapy using tumor suppressor gene is considered as a potent candidate for new therapeutic approaches that can improve efficacy and reduce complications. The purpose of this research is to identify the role of adenoviral vector to transfer HCCS-1 tumor suppressor gene in oral cancer cells and to find out whether there is a possibility for it to serve in the field of gene therapy. The human SCC-25 cell line was used for transfection. To determine the efficiency of the adenovirus as a gene delivery vector cell line was transduced with LacZ gene and analysed with X-gal staining. Northern blot was performed to confirm the tranfection with HSCC-1 gene and cell viability was assessed by cell cytotoxicity assay. We had successfully construct the recombinant HSCC-1 adenovirus(Ad5CMV-HCCS-1). DNA extracted from Ad5CMV-HCCS-1 revealed HCCS-1 gene is incorporated. The transduction efficiencies were over than 50% of SCC-25 cells with a MOI of 2 and over 95% with a MOI of 50. Northern blot analysis showed that a single 0.6kb mRNA transcript was expressed in Ad5CMV-HCCS-1 transduced SCC-25 cells. There was no or very low transcription HCCS-1 mRNA in wild and Ad5CMV-LacZ transduced SCC-25 cells. Cells transduced with Ad5CMV-HCCS-1 showed significant growth inhibition. By day 6, Ad5CMV-HCCS-1 treated cell count was decreased to 30% of mock-infected cells, while that of Ad5CMV-LacZ treated cells was 90% of mock-infected cells (p<0.05). Finally, these result suggest that the Ad5CMV-HCCS-1 has potential as a gene therapy tool for oral cancer.

• Journal of Korean Neurosurgical Society
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• v.29 no.4
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• pp.471-476
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• 2000

Objective : p16/INK4a, a kind of tumor suppressor genes, encodes a specific inhibitor of the cyclin D-dependent kinases CDK4 and CDK6. This prevents the association of CDK4 with cyclin D1, and subsequently inhibits phosphorylation of retinoblastoma tumor suppressor protein(pRb), thus preventing exit from the G1 phase. According to previous reports, over 50% of glioma tissue and 80% of glioma cell lines have been demonstrated inactivation of p16/INK4a gene. The purpose of this study was to determine whether recombinant adenovirus-p16 virus is a suitable candidate for gene replacement therapy in cases of glioma. Methods : Three human glioma cell lines(U251MG, U87MG and U373MG) that express mutant p16 protein were used. Replication-deficient adenovirus was utilized as an expression vector to transfer exogenous p16 cDNA into the cells ; control cells were infected with the Ad-${\beta}$-gal expressing ${\beta}$-galactosidase. To monitor gene transfer and the expression of exogenous genes, we used Western Blotting analysis. Flow cytometry studies of cellular DNA content were performed to determine the cell cycle phenotype of the glioma cells before and after treatment. Results : We showed here that restoration of p16/INK4a expression in p16 negative U87MG, U251MG and partially deleted U373MG by Ad-CMV-p16 induced growth suppression in vitro. Flow cytometric study revealed that Ad-CMV-p16 infected U87MG cells were arrested during the G0-G1 phase of the cell cycle. Expression of p16 transferred by Ad-CMV-p16 in glioma cells was highly efficient and maintained for more than seven days. Conclusions : Our results suggest that Ad-CMV-p16 gene therapy strategy is potentially useful and warrants further clinical investigation for the treatment of gliomas.

• Pediatric Infection and Vaccine
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• v.21 no.3
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• pp.207-213
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• 2014

Purpose: This study aimed to investigate the association between respiratory virus infection and pneumococcal colonization in children. Methods: From May 2009 to June 2010, nasopharyngeal (NP) aspirates were obtained from patients under 18 years old who visited Seoul National University Children's Hospital for respiratory symptoms. NP samples were used to detect respiratory viruses (influenza virus A and B, parainfluenza virus 1, 2 and 3, respiratory syncytial virus A and B, adenovirus, rhinovirus A/B, human metapneumovirus, human coronavirus 229E/NL63 and OC43/HKU1) by RT-PCR and pneumococcus by culture. Results: Median age of the patients was 27 months old. A total of 1,367 NP aspirates were tested for respiratory viruses and pneumococcus. Pneumococcus was isolated from 228 (16.7%) of samples and respiratory viruses were detected from 731 (53.5%). Common viruses were rhinovirus (18.4%), respiratory syncytial virus (RSV) A (10.6%), adenovirus (6.9%), influenza virus A (6.8%). Pneumococcal isolation rate was significantly higher in the cases of positive virus detection than negative detection [21.3% (156/731) vs. 11.3% (72/636), P <0.001]. For individual viruses, pneumococcal isolation rate was positively associated with detection of influenza virus A [24.7% (23/93) vs 16.1% (205/1274), P=0.001], RSV A [28.3% (41/145) vs 15.3% (187/1222), P=0.001], RSV B [31.3% (10/32) vs 16.3% (218/1335), P=0.042], rhinovirus A/B [22.6% (57/252) vs 15.3% (171/1115), P=0.010]. Conclusion: The study revealed that pneumococcal isolation from NP aspirates is related with respiratory virus detection. The result of this study could be used to investigate how respiratory viruses and pneumococcus cause clinical diseases.

• Journal of Life Science
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• v.30 no.3
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• pp.221-229
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• 2020

Distylium racemosum is an evergreen tree growing wild on Jeju Island, which has been reported to exert biological activity. Obesity is induced by genetic, metabolic, environmental, and other factors. Among these, certain bacterial and viral infections have been shown to cause obesity, which is known as infectobesity. Human adenovirus (HAdV)-36 is one of the viruses that are known to cause infectobesity in humans. Unlike extensive research on preventing obesity and developing anti-obesity drugs, little research has been conducted specifically on the prevention and treatment of infectobesity. Therefore, this study aimed to evaluate the effects of phytochemicals from D. racemosum on the replication of HAdV-36. A549 cells infected with HAdV-36 were treated with an ethyl acetate fraction of a D. racemosum leaf extract (DRE), and the virus titer was calculated based on the hemagglutination (HA) titer. The results showed a concentration-dependent inhibitory effect of DRE treatment on HA titers. DRE treatment was also found to inhibit the cytopathic effects of the virus and the expression of viral genes. Quercitrin was identified as the constituent of DRE exerting an inhibitory effect on HAdV-36 replication. This study shows that DRE can be used as a candidate substance for the development of treatment for HAdV-36 infections. In addition, this study provides a basis to further investigate DRE for the development of anti-infectobesity medication.

• Clinical and Experimental Pediatrics
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• v.52 no.3
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• pp.269-276
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• 2009

Viruses are the most common cause of lower respiratory tract infections (LRTIs) in infants and young children and are a major public health problem in this age group. Viruses were identified in 54.9-70.4% of hospitalized infants and children with LRTIs in Korea. The viral pathogens identified included respiratory syncytial virus (RSV) A and RSV B, influenza (Inf) A, Inf B, parainfluenza (PIV)1, PIV2, human bocavirus (hBoV), human rhinovirus (hRV), adenovirus (ADV), human metapneumovirus (hMPV), human coronavirus (hCoV)-OC 43, hCoV-229E, hCoV-NL63, hCoV-HKU1, and human enterovirus (hEV). Coinfections with ${\geq}$2 viruses were observed in 11.5-22.8% of children. The occurrence of LRTIs was the highest in the first year of life. The specific viruses are frequently associated with specific clinical syndromes of LRTIs. LRTIs caused by RSV were predominant among younger infants. hRV accounted for a larger proportion of LRTIs in young infants than ADV and hBoV. hMPV was frequently detected in children >24 months old. The number of hMPV infections peaked between February and May, whereas hRV was detected throughout the year. Thus far, hCoV is a less common respiratory pathogen in cases of ALRI and URI in Korean children.

• Journal of Microbiology and Biotechnology
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• v.18 no.6
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• pp.1156-1163
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• 2008

We performed RT-nested PCR to study the distribution of human enteric viruses in urban rivers in Korea. During 2002-2003, water samples were collected from four rivers in Gyeonggi Province, South Korea. Among 58 samples, 45 (77.6%), 32 (55.2%), 12 (20.7%), 2 (3.4%), 4 (6.9%), and 4 (6.9%) showed positive results with adenoviruses (AdVs), enteroviruses (EVs), reoviruses (ReVs), hepatitis A viruses (HAVs), rotaviruses (RoVs), and sapoviruses (SVs), respectively. According to the binary logistic regression model, the occurrence of each enteric virus, except ReVs and HAVs, was not statistically correlated with the water temperature and levels of fecal coliforms (P<0.05). AdVs were most often detected; only 4 samples (6.9%) were negative for AdVs while positive for other enteric viruses in the studied sites. Our results indicated that monitoring human enteric viruses is necessary to improve microbial quality, and that AdVs detection by PCR can be a useful index for the presence of other enteric viruses in aquatic environments.

• Journal of Microbiology and Biotechnology
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• v.23 no.2
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• pp.267-273
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• 2013

Multiplex RT-PCR was used to detect respiratory viruses in 5,318 clinical samples referred to the laboratory of a tertiary teaching hospital from December 2006 to November 2010. The acquired data were analyzed with respect to types, ratio, and co-infection trends of infected respiratory viruses. Trends in respiratory viral infection according to sex, age, and period of infection were also analyzed. Of the 5,318 submitted clinical samples, 3,350 (63.0%) specimens were positive for at least one respiratory virus. The infection rates were 15.8% for human rhinovirus, 14.4% for human respiratory syncytial virus A, 9.7% for human respiratory syncytial virus B, 10.1% for human adenovirus, 5.4% for influenza A virus, 1.7% for influenza B virus, 4.7% for human metapneumovirus, 2.3% for human coronavirus OC43, 1.9% for human coronavirus 229E/NL63, 3.7% for human parainfluenza virus (HPIV)-1, 1.1% for HPIV-2, and 5.3% for HPIV-3. The co-infection analysis showed 17.1% of double infections, 1.8% of triple infections. The median age of virus-positive patients was 1.3 years old, and the 91.5% of virus-positive patients were under 10 years old. Human respiratory syncytial virus was the most common virus in children < 5 years of age and the influenza A virus was most prevalent virus in children over 5 years of age. These results help in elucidating the tendency of respiratory viral infections.