• Title/Summary/Keyword: 역전사 중합효소연쇄반응

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Expression of Luteinizing Hormone (LH) Subunit Genes in the Rat Ovary (흰쥐 난소에서의 Luteinizing Hormone (LH) Subunit 유전자 발현)

  • Lee, Sung-Ho
    • Clinical and Experimental Reproductive Medicine
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    • v.25 no.2
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    • pp.199-205
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    • 1998
  • The present study was performed to analyze the expression of LH genes in the rat ovary. Expression of LH subunit genes in the rat ovary was demonstrated by amplification of ovarian RNA by RT-PCR. The ovarian $LH_\beta$ transcripts contained at least two parts of the published cDNA structure, the pituitary exons 1, 2 and 3 and the part of testicular ex on 1 in the major trancripts form in rat testis. Using RIA, significant amount of LH-like molecules were detected in crude ovarian extracts, and the competition curves with increasing amount of tissue extracts were parallel with those of standard peptide, indicating that the ovarian immunoreactive LH-like material is similar to authentic pituitary LH molecule. The administration of PMSG to immature rats resulted in a sharp decrease of the ovarian LH contents after 24 h post-injection. In conclusion, these findings demonstrate that genes for LH subunits are expressed in the rat ovary, and suggest that LH can playa central role in regulation of female reproduction with both endocrine (by pituitary LH) and auto- and/or para-crine (by ovarian LH) manner.

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IFN-${\gamma}$mRNA Expression in Tuberculous Pleural Lymphocytes After in vitro Stimulation with M. tuberculosis Antigens (결핵균 항원 자극에 의한 결핵성 흉수 림프구의 IFN-${\gamma}$ mRNA 발현)

  • Park, Jae Seuk;Kim, Youn Seup;Jee, Young Koo;Lee, Kye Young
    • Tuberculosis and Respiratory Diseases
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    • v.57 no.1
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    • pp.25-31
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    • 2004
  • Background : IFN-${\gamma}$ is the main effector mediator of the host immune response against Mycobacterium tuberculosis. Evaluating the IFN-${\gamma}$ gene expression in response to M. tuberculosis antigens may help in elucidating the host defense mechanism against M. tuberculosis and in the development of a vaccine. Methods : The IFN-${\gamma}$ mRNA expression in the lymphocytes obtained from pleural effusions from tuberculous pleurisy patients (TB-PLC) after in vitro stimulation with whole cell M. tuberculosis(H37Rv), purified protein derivatives(PPD), man-lipoarabinamman (man-LAM), ara-LAM and Antigen 85B(Ag85B) were evaluated. The degree of IFN-${\gamma}$ mRNA expression was determined by a semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR) method. Results : M. tuberculosis induced the expression of IFN-${\gamma}$ mRNA in the TB-PLC in time and dose dependent manners. The PPD and Ag85B induced high levels of IFN-${\gamma}$ mRNA expression in the TB-PLC. However, man-LAM inhibited IFN-${\gamma}$ mRNA expression in the TB-PLC, while ara-LAM did not. Conclusion : IFN-${\gamma}$ mRNA expression in TB-PLC is stimulated by PPD and Ag85B, but inhibited by man-LAM.

Detection of Cymbidium Mosaic Virus and Odontoglossum Ringspot Virus in Seed-Derived Plantlets of Phalaenopsis Imported by One-Step RT-PCR (One-Step RT-PCR 방법에 의한 수입 호접란묘의 심비디움 모자이크 바이러스와 오돈토글로섬 윤문 바이러스의 검정)

  • Yun, Jong Sun;Hong, Eui Yon;Kim, Ik Hwan;Yun, Tae;Kim, Tae Su;Paek, Kee Yoeup
    • Horticultural Science & Technology
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    • v.18 no.4
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    • pp.513-517
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    • 2000
  • This experiment was carried out to detect the cymbidium mosaic virus (CymMV) and the odontoglossum ringspot virus (ORSV) in the seed-derived plantlets of Phalaenopsis imported from Taiwan by one-step reverse transcription-polymerase chain reaction (RT-PCR). Simple and rapid crude plant extracts for RT-PCR were prepared. The reverse transcription step was performed at $42^{\circ}C$ for 45 min and the following thermal cycling scheme was used for 36 reaction cycles: template predenaturation at $96^{\circ}C$ for 2 min, template denaturation at $96^{\circ}C$ for 30 s, primer annealing at $60^{\circ}C$ for 30 s, and DNA synthesis at $72^{\circ}C$ for 1 min. Of the 40 seed-derived plantlets of Phalaenopsis imported from Taiwan, all of them were infected with CymMV, but ORSV was not detected.

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The effect of blasting and anodizing-combined treatment of implant surface on response of osteoblast-like cell (분사처리 후 양극산화 처리한 임플란트 표면이 골모 유사 세포의 반응에 미치는 영향)

  • Seo, Bo-Yong;Kim, Young-Min;Choi, Jae-Won;Yun, Mi-Jung;Jeon, Young-Chan;Jeong, Chang-Mo;Kim, Gyu-Cheon;Huh, Jung-Bo
    • The Journal of Korean Academy of Prosthodontics
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    • v.53 no.1
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    • pp.9-18
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    • 2015
  • Purpose: The purpose of this study is to examine characteristics of implant surface with RBM and anodizing treatments, and to evaluate the responses of osteoblast-like cell (MG-63 cell). Materials and methods: Grade IV titanium disks were fabricated (Diameter 10 mm, thickness 3 mm). Anodizing treatment (ASD) group, RBM and anodizing treatment (RBM/ASD) group, control (machined surface) group were divided. In this study, osteoblast-like cell was used for experiments. The experiments consist of surface characteristics evaluation by FE-SEM images, energy dispersive spectroscopy and stereo-SEM. In order to evaluate cell adhesion evaluation by crystal violet assay and observe cells form by confocal laser microscopy. To assess cell proliferation by XTT assay, cell differentiation by RT-PCR and mineralization by Alizarin red S stain assay. ELISA analyzer was used for Quantitative evaluation. Comparative analysis was run by one-way ANOVA (SPSS version 18.0). Differences were considered statistically significant at P<.05. Results: In ASD group and RBM/ASD group, the surface shape of the crater was observed and components of oxygen and phosphate ions in comparison with the control group were detected. The surface average roughness was obtained $0.08{\pm}0.04{\mu}m$ in the control group, $0.52{\pm}0.14{\mu}m$ in ASD group and $1.45{\pm}0.25{\mu}m$ in RBM/ASD group. In cell response experiments, ASD group and RBM/ASD group were significantly higher values than control group in cell adhesion and mineralization phase, control group was the highest values in the proliferative phase. In RT-PCR experiments, RBM/ASD group was showed higher ALP activity than other groups. RBM/ASD group in comparison with ASD group was significantly higher value for cell adhesion and proliferation phase. Conclusion: In the limitation of this study, we are concluded that the surface treatment with RBM/ASD seems more effective than ASD alone or machined surface on cellular response.

INHIBITION OF GLUCAN SYNTHESIS RELATED GENE EXPRESSION OF STREPTOCOCCUS MUTANS BY XYLITOL TREATMENT (자일리톨 섭취에 따른 Streptococcus mutans의 글루칸 생성관련 유전자 발현 억제효과)

  • Kim, Ji-Hye;Lee, Young-Eun;Ahn, Sang-Hun;Choi, Youn-Hee;Nam, Soon-Heyun;Song, Keun-Bae
    • Journal of the korean academy of Pediatric Dentistry
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    • v.36 no.4
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    • pp.531-538
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    • 2009
  • Xylitol has the ability to reduce the adherence of Streptococcus mutans(S. mutans), which can make it easier to remove plaque, decrease acid production and inhibit dental caries. There are few reports on the effects of xylitol on the expression of the virulence related genes in S. mutans. This study examined the inhibitory effect of chewing gum containing xylitol on glucan synthesis related gene expression of S. mutans. Participants were voluntarily recruited for a women's oral health prevention program, classified into two groups(a control and a xylitol group), and then followed for 2 years. Twenty salivary samples were randomly selected from each group. Colony count and real-time reverse transcription polymerase chain reaction were used to analyze the characteristics of S. mutans. The following results were obtained: The S. mutans counts decreased steadily in the xylitol group over the study period(p<0.05). The expression of the virulence related genes (gtfB, gtfC and gtfD) was significantly lower in the xylitol group than in the control groups (p<0.05). In conclusion, these results suggest that chewing xylitol gum for a long period of time may reduce the expression of the genes associated with S. mutans virulence, which can result in a decrease growth of S. mutans colonies as a result.

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Cardiopulmonary bypass -induced Gene expressions of proinflammatory cytokines and chemokines (체외순환에서 전염증성 사이토카인 및 케모카인 유전자 발현 특성에 대한 연구)

  • Yu, Jae-Hyeon;Jo, Eun-Kyeong;Park, Jeong-Kyu;Kang, Shin-Kwang;Na, Myung-Hoon;Lim, Seung-Pyung;Lee, Young
    • Journal of Chest Surgery
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    • v.35 no.2
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    • pp.118-126
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    • 2002
  • 배경: 체외순환에 의해 생성되는 염증매개체는 소아 환자에서 술 후 다기관 기능부전과 연관이 있다. 본 연구에서는 선천성 신장질환 소아환자에서 체외순환에 의한 전염증성 사이토카인과 케모카인의 유전자 발현 특성에 대해 알아보고자 하였다. 대상 및 방법: 개심술을 시행한 18명의 소아 환자의 요골 동맥에서 마취유도 후(기준치), 체외순환(0시간), 체외순환 2시간 후 24시간 후, 48시간 후에 혈액을 채취하였다. 모든 환자에서 인터루킨-1알파, 인터루킨-1베타 인터루킨-6, 인터루킨-8, 종양괴사자인자-알파, 인터루킨-15,인터페론 감마의 mRNA의 유전자 발현 정도를 반정량적으로 역전사 중합효소 연쇄반응으로 측정하였다. 6명의 환자에서 인터루킨-6단백치는효소결합면역흡착검사로 측정하였다. 결과: 전신적인 인터루킨-6 mRNA와 비슷한 양상을 보였으나 최고치는 인터루킨-6보다 낮은 값을 보였다. 인터루킨-1알파와 인터루킨-1베타의 mRNA의 발현은 체외순환 2시간 후에 최고치를 나타내었고 체외순환 2시간 후에 최고치를 나타내었고 체외순환 48시간 후에 감소하였다. 종양괴사자인자-알파는 체외 순환 24시간 후에 최고치를 나타내었고 체외순환 48시간 후에 감소하였다. 인터루킨-15 mRNA 발현은 체외순환 전후와 비교하여 유의한 변화가 없었다. 인터페론-감마는 시간이 지남에 따라 감소하였다. 결론: 선천성 심장질환으로 개심술을 시행한 소아환자의 혈청 내 인터루킨-6, 인터루킨-8, 인터루킨-1알파, 인터루킨-베타, 종양괴사자인자-알파의 유전자 발현은 체외 순환 전후로 유의한 변화를 보였다. 인터루킨-15는 유의한 변화가 없었고 인터페론-감마는 반대 양상의 변화를 보였다. 체외순환 후 전염증성 사이토카인과 케모카인의 높은 혈중 농도는 술 후 조직 손상과 연관되리라 생각한다.

Study on Laboratory Diagnosis of the Ebola Virus and Its Current Trends (에볼라 바이러스 진단법과 개발 동향에 관한 고찰 연구)

  • Jeong, Hye Seon;Kang, Yun-Jung
    • Korean Journal of Clinical Laboratory Science
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    • v.47 no.3
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    • pp.105-111
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    • 2015
  • In late December 2013, the Ebola virus emerged from West Africa. The outbreak started in Guinea and rapidly spread to Liberia and Sierra Leone. Initially, the virus is spread to the human population after contact with infected wildlife and then spread person-to-person through direct contact with body fluids such as blood, sweat, urine, semen, and breast milk. The Ebola virus infects endothelial cells, mononuclear phagocytes and hepatocytes. It causes massive damage to internal tissues and organs, such as blood vessels and the liver, and ultimately death. Most tests for the virus RNA rely on a technology called reverse-transcriptase polymerase chain reaction (RT-PCR). While this method is highly sensitive, it is also expensive, requiring skilled scientists, and delicate power supplies. The strip analytical technique (enzyme-linked immunosorbent assay or ELISA) detects antigens or antibodies to the Ebola virus. This test is cheap and does not require electricity or refrigeration. Despite ongoing efforts directed at experimental treatments and vaccine development, current medical work on the Ebola viral disease is largely limited to supportive therapy. Thus, rapid and reliable diagnoses of the Ebola virus are critically important for patient management, infections, prevention, and control measures.

The effect of progressive tensional force on mRNA expression of osteoprotegerin and receptor activator of nuclear factor ${\kappa}B$ ligand in the human periodontal ligament cell (기계적 자극이 치주인대 세포의 osteoprotegerin과 receptor activator of nuclear factor ${\kappa}B$ ligand mRNA 발현에 미치는 영향)

  • Lee, Kie-Joo;Lee, Syng-Ill;Hwang, Chung-Ju;Ohk, Seung-Ho;Tian, Yu-Shin
    • The korean journal of orthodontics
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    • v.35 no.4 s.111
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    • pp.262-274
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    • 2005
  • Tooth movement is a result of mutual physiologic responses between the periodontal ligament and alveolar bone stimulated by mechanical strain. The PDL cell and osteoblast are known to have an influence on bone formation by controlling collagen synthesis and alkaline phosphatase activation. Moreover. recent studies have shown that the PDL cell and osteoblast release osteoprotegerin (OPG) and the receptor activator of nuclear factor ぉ ligand (RANKL) to control the level of osteoclast differentiation and activation which in turn influences bone resorption. In this study. progressively increased, continuous tensional force was applied to PDL cells. The objective was to find out which kind of biochemical reactions occur after tensional force application and to illuminate the alveolar bone resorption and apposition mechanism. Continuous and progressively increased tensile force was applied to PDL cells cultured on a petriperm dish with a flexible membrane The amount of $PGE_2$ and ALP synthesis were measured after 1, 3, 0 and 12 hours of force application. Secondly RT-PCR analysis was carried out for OPG and RANKL which control osteoclast differentiation and MMP-1 -8, -9, -13 aud TIMP-1 which regulate the resolution of collagen and resorption of the osteoid layer According to the results. we concluded that progressively increased, concluded force application to human PDL cells reduces $PGE_2$ synthesis, and increases OPG mRNA expression.

Effects of Mineral Trioxide Aggregate on the Proliferation and Differentiation of Human Dental Pulp Stromal Cells from Permanent and Deciduous Teeth (Mineral trioxide aggregate가 유치 및 영구치의 치수기질세포 증식 및 분화에 미치는 영향)

  • Kim, Seunghye;Jeon, Mijeong;Shin, Dong Min;Lee, Jae Ho;Song, Je Seon
    • Journal of the korean academy of Pediatric Dentistry
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    • v.40 no.3
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    • pp.185-193
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    • 2013
  • Mineral trioxide aggregate (MTA) has recently been used as a pulpotomy medicament for primary molars. The aim of this study was to evaluate and compare the proliferation and differentiation potential of dental pulp stromal cells of permanent teeth and deciduous teeth cultured on MTA-coated surface. Human dental pulp stromal cells were obtained from human permanent premolars and deciduous teeth and cultured on MTA-coated culture plates. The cells were subjected to proliferation assay and cell cycle analysis. Their differentiation potential was evaluated by analysing changes in the mRNA expressions of runt-related transcriptional factor 2 (Runx2) and alkaline phosphatase (ALP). Morphological changes of cells in direct contact with MTA were observed using scanning electron microscopy (SEM). The proliferation rates, distribution of cell cycles and mRNA expression patterns of Runx2 and ALP were similar in both types of pulpal cells. SEM observations revealed that both types changed into more dendrite-like cells. On the surface of MTA, human dental pulp stromal cells from deciduous and permanent teeth were able to both proliferate and differentiate into cells that induce mineralization. MTA is suitable as a biocompatible pulpotomy medicament for primary teeth.

Distribution of Disease Severity and Group A Rotavirus Genotypes (VP7 & VP4) in Children with Acute Rotavirus Gastroenteritis (급성 로타바이러스 장염 소아에서 질환의 중증도와 로타바이러스 VP7 & VP4 유전형의 분포에 대한 연구)

  • Oh, Hyun-Ju;Kang, Hyun-Sik;Kang, Ki-Soo;Kim, Youn-Woo;Hong, Jung-Yun;Shin, Kyung-Sue;Lee, Jin-Sook;Lee, Su-Hyun;Lee, Keun-Hwa;Cho, Moon-Je;Suh, Dong-In
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.14 no.2
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    • pp.148-154
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    • 2011
  • Purpose: We aimed to study the distribution of rotavirus genotypes (VP7 and VP4) and disease severity of rotavirus gastroenteritis prevalent in our community. Methods: Stool samples were collected from 156 children who were hospitalized with rotavirus gastroenteritis from December 2007 to June 2008. The disease severity of all patients was scored using the Vesikari scale. After extraction of ds-RNA of the rotavirus, cDNA synthesis using reverse transcription and polymerase chain reaction (RT-PCR) and multiplex PCR was performed. Following this, the final identification of genotypes was performed. Results: Of the 156 samples, VP7(G) and VP4(P) genotypes were identified in 147 (94.2%) and 140 (89.7%) samples, respectively. G1 (116 of 147 samples; 78.9%) and P[8] (137 of 140 samples; 97.9%) were the most prevalent, respectively. Of the 138 samples identified of combination types of VP7 and VP4, G1P[8] (111 samples; 80.4%) was the most prevalent. Other combination types varied with very low distribution rates. 9.4% of genotypes were not included in the new vaccines. The disease severity score was $11.8{\pm}3.3$ ($mean{\pm}2SD$). The distribution of disease severity was mild or moderate in 37.8% and severe in 62.2% of patients. Conclusion: The most prevalent genotype combination of rotavirus was G1P[8] and genotypes not included in the vaccines represented 9.4% in our community. Disease severity distribution of hospitalized children with rotavirus gastroenteritis was higher in the severe than in the mild and moderate categories.