• Journal of Periodontal and Implant Science
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• v.40 no.1
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• pp.39-44
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• 2010

Purpose: Bone tissues for clinical application can be improved by studies on osteoblast differentiation. Runx2 is known to be an important transcription factor for osteoblast differentiation. However, bone morphogenetic protein (BMP)-2 treatment to stimulate Runx2 is not sufficient to acquire enough bone formation in osteoblasts. Therefore, it is necessary to find other regulatory factors which can improve the transcriptional activity of Runx2. The erythroblast transformation-specific (ETS) transcription factor family is reported to be involved in various aspects of cellular proliferation and differentiation. Methods: We have noticed that the promoters of osteoblast differentiation markers such as alkaline phosphatase (Alp), osteopontin (Opn), and osteocalcin (Oc) contain Ets binding sequences which are also close to Runx2 binding elements. Luciferase assays were performed to measure the promoter activities of these osteoblast differentiation markers after the transfection of Runx2, myeloid Elf-1-like factor (MEF), and Runxs+MEF. Reverse-transcription polymerase chain reaction was also done to check the mRNA levels of Opn after Runx2 and MEF transfection into rat osteoblast (ROS) cells. Results: We have found that MEF, an Ets transcription factor, increased the transcriptional activities of Alp, Opn, and Oc. The addition of Runx2 resulted in the 2- to 6-fold increase of the activities. This means that these two transcription factors have a synergistic effect on the osteoblast differentiation markers. Furthermore, early introduction of these two Runx2 and MEF factors significantly elevated the expression of the Opn mRNA levels in ROS cells. We also showed that Runx2 and MEF proteins physically interact with each other. Conclusions: Runx2 interacts with MEF proteins and binds to the promoters of the osteoblast markers such as Opn nearby MEF to increase its transcriptional activity. Our results also imply that osteoblast differentiation and bone formation can be increased by activating MEF to elicit the synergistic effect of Runx2 and MEF.

• Journal of Chest Surgery
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• v.47 no.4
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• pp.378-383
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• 2014

Background: Brother of the regulator of imprinted sites (BORIS) is a putative new oncogene that is classified as a cancer germline gene; however, its role in the development of cancer is unclear. This study investigated the expression of BORIS in lung cancer and its clinical implications. Methods: The expression of BORIS messenger ribonucleic acid (mRNA) in the sputum of 100 patients with lung cancer (50 with squamous cell carcinoma, 36 with adenocarcinoma, and 14 with small-cell carcinoma) was evaluated by reverse transcription polymerase chain reaction. Results: The overall expression rate of BORIS in patients with lung cancer was 36.0%: 19 of 50 squamous cell carcinomas (38.0%), 13 of 36 adenocarcinomas (36.1%), and 4 of 14 (28.6%) small-cell carcinomas. There was no significant difference in the BORIS expression according to age, gender, or histologic type. However, the mRNA expression of BORIS was significantly related to the pathologic cancer stage (p=0.004) and lymph node metastasis (p=0.001). The expression of the melanoma antigen gene family A1-6 was not associated with the expression of BORIS. Conclusion: Our results suggest that the expression of BORIS might be a negative prognostic factor in lung cancers and implicate BORIS as a molecular target for immunotherapy.

• Journal of the East Asian Society of Dietary Life
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• v.7 no.4
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• pp.484-490
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• 1997

In an attempt to investigate the effects of white ginseng on the proliferation and the nitric oxide(NO) secretion of mouse peritoneal macrophages, which are the first mai or defense phagocytes in the immune system, the studies have been carried out. In the macrophage proliferation assay using the $^3$H-thymidine incorporation, the total saponin or Ginsenoside Rb$_2$ were added to the medium at the concentration of 0 to 256$\mu\textrm{g}$/$m\ell$. DNA synthesis of the macrophage was increased at 64$\mu\textrm{g}$/$m\ell$ of total saponin and either 16$\mu\textrm{g}$/$m\ell$ or 64$\mu\textrm{g}$/$m\ell$ of Ginsenoside Rb$_2$, respectively. Also, the effect o(white ginseng on the nitric oxide secretion of the macrophages was investigated. The addition of either total saponin or Ginsenoside Rb$_2$ at the concentration of 20$\mu\textrm{g}$/$m\ell$ significantly increased the secretion of NO from the macrophages. The nitric oxide synthase (NOS) gene expression which is responsible for the synthesis of the nitric oxide has been studied using reverse transcription polymerase chain reaction. In RT-PCR, the $\beta$-actin and nos gene expression have been analyzed. 20$\mu\textrm{g}$/$m\ell$ of either total saponin or Ginsenoside Rb$_2$ increased nos gene expression of the macrophages.

• Journal of the korean academy of Pediatric Dentistry
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• v.42 no.4
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• pp.312-318
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• 2015

The aim of this study was to investigate the possibility of the supernumerary teeth for the stem cell source in dentistry. The Real Time Quantitative Reverse Transcription Polymerase Chain Reaction (Real Time qRT-PCR) method was used to evaluate the differentiation toward the odontoblast of the supernumerary dental pulp stem cells (sDPSCs). Supernumerary dental pulp stem cells were obtained from 3 children (2 males and 1 female, age 7 to 9) diagnosed that the eruption of permanent teeth was disturbed by supernumerary teeth. The common genes for odontoblasts are alkaline phosphatase (ALP), osteocalcin (OC), osteonectin (ON), dentin matrix acidic phosphoprotein 1 (DMP-1), dentin sialophosphoprotein (DSPP). The sDPSCs were treated for 0 days, 8 days and 14 days with additives and then Real Time qRT-PCR was performed in intervals of 0 days, 8 days and 14 days. The alizarin-red solution staining was performed to visualize the stained color for the degree of calcification at 7 days, 14 days, 21 days and 28 days after treating additives to the sDPSCs. From the result of the Real Time qRT-PCR, the manifestation exhibit maximum value at 8 days after additive treatment and shifted to a decrease trend at 14 days. Alizarin-red solution staining exhibit light results at 7 days after staining and generalized dark result at 14 days. Consequently, in studies with sDPSCs, appropriate treatment time of additives for Real Time qRT-PCR is 8 days. Also, a suitable period of Alizarin-red solution staining is 14 days.

• Development and Reproduction
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• v.4 no.2
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• pp.231-236
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• 2000

Recent studies have clearly shown that the expression of genes for gonadotropin-releasing hormone (GnRH) and its receptor in the rat reproductive organs including ovary, testis, placenta uterus and mammary gland. Moreover, luteinizing hormone (LH) classically known to be a main target product of GnRH in anterior pituitary has been found in rat gonads. These findings suggested the presence of local circuit composed of GnRH and LH in the rat gonads. The present study was undertaken to elucidate whether the genes for LH and its receptor are expressed in rat mammary gland. Expression of LH and its receptor genes in the rat mammary gland was demonstrated by reverse transcription-polymerase chain reaction (RT-PCR) and specific LH radioimmunoassay (RIA). The LH${\beta}$ transcripts in the mammary gland from cycling rats contained the pituitary type of LH${\beta}$ exons 1~3 encoding the entire LH${\beta}$ polypeptide but lacked the rat testis-specific LH${\beta}$ exon(s). Presence of ${\alpha}$ -subunit transcripts in the rat mammary gland were determined by RT-PCR. The cDNA fragments encoding exons 2~7 of rat LH receptor transcripts were amplified in both rat ovary and mammary gland samples. We could detect the GnRH expression in mammary gland from cycling virgin rats, and this result disagreed with previous report that mammary GnRH expression is occured in lactating rats only. Considerable amounts of immunoreactive LH molecules with good RIA parallelism in standard curve were detected in crude extracts from the rat mammary gland, indicating that the immunoreactive LH materials in the gland might be identical to authentic pituitary LH. To our knowledge, the present study demonstrated for the first time the expression of LH subunits and LH receptor in the rat mammary gland. Our findings suggested that the mammary gland might be the novel source and target of LH and the mammary LH could be act as a local regulator with auto-and/or paracrine manner under the regulation of local GnRH.

• The Journal of the Korean Society for Microbiology
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• v.34 no.5
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• pp.453-459
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• 1999

Astrovirus is frequently associated with diarrhea in children. It can not be readily isolated by cell culture, and an electronmicroscope is usually used for detection of this agent. Recently in 1995 a combined method of reverse transcription-polymerase chain reaction (RT-PCR) was designed for easier detection of astrovirus, which is based on the conserved sequence in 3'-end of genomes of the 7 known serotypes of human astrovirus. As of yet there has not been any report of astrovirus data in Korea using the RT-PCR methods. The purpose of this study was to detect astrovirus incidence, severity of symptoms, seasonal variation and co infection rate with rotavirus in Korean children inpatients with diarrhea. Fecal specimens from 61 young children hospitalized with gasteroenteritis Korea from Jan. 1996 through Mar. 1997. They were examined for astroviurs infection by RT-PCR method. Results are as follows:1. Astrovirus was detected at 9.8% (6/61) from fecal specimens of children with severe diarrhea by EIA using monoclonal antibody coated plates. 2. Astorvirus was detected at 29.5% (18/61) from fecal specimens of children with severe diarrhea by RT-PCR. 3. The age of the 18 children affected by astrovirus ranged from 2 monthes to 7 years with mean of 3.0 years. 4. Mean hospital stay of the 18 children was 6.1 days. 5. Five (27.8%) astrovirus RT-PCR positive strains were confirmed in November and in December, respectively out of 18 specimens in total. 6. Astrovirus coinfection with rotavirus type G1 was confirmed in 15/16 specimens (93.8%), and with type G2 was in 1/16 specimens (6.3%).

• Research in Plant Disease
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• v.25 no.4
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• pp.233-236
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• 2019

Citrus vein enation virus (CVEV), which was regulated as a quarantine virus in Korea, was firstly found on Jeju Island in 2017. In February 2018, a survey was carried out to determine the distribution of CVEV in the main commercial areas growing Citrus spp. and Poncirus trifoliata. The survey was performed at 203 groves in the southern Korean Peninsula and Jeju Island. CVEV infection was determined by reverse transcription polymerase chain reaction detection and sequencing. The coat protein (CP) gene sequences obtained from the CVEV-infected samples showed high similarities (more than 98%) to the previously reported CVEV CP sequences. In summary, CVEV was detected in 136 groves (67%), in which 85.4% of Citrus junos and 77.8% of Citrus unshiu were infected by CVEV. In Jeju Island, the infection rate of CVEV was relatively higher (90.6%). Our result revealed that CVEV has spread widely in Citrus and Poncirus in Korea. Based on the result, the Korean quarantine agency decide to exclude CVEV from quarantine in Korea.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.43 no.1
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• pp.19-26
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• 2017

This study investigated the anti-inflammatory activities and cell viability of Amelanchier asiatica (A. asiatica) 70% ethanol extracts against RAW 264.7 cells induced by lipopolysaccharide (LPS). Cell toxicity test on macrophage cells (RAW 264.7) was performed by 3-[4,5-dimethyl-thiazol-2-yl]-2, 5-diphenyl-tetrazoliumbromide (MTT) assay and results showed 96% cell viability at $1,000{\mu}g/mL$ concentration. Anti-inflammatory activity was examined via the inhibitory tests on the production of LPS induced NO in RAW 264.7 cells by Griess assay. The result showed that the extract inhibited NO production in concentration dependent manner. The iNOS and COX-2 protein expression inhibitory effects were confirmed by western blot and by reverse transcription-polymerase chain reaction (RT-PCR). From the former they were decreased by 84.3%, 56.2% at $500{\mu}g/mL$ concentration, respectively, and from the latter decreased by 89.8%, 84.9% at $500{\mu}g/mL$, respectively. In conclusion, this study showed the anti-inflammatory effects of A. asiatica extracts. Thus, this could be applied to an anti-inflammatory agent.

• Journal of Korean Medicine Rehabilitation
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• v.24 no.2
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• pp.1-13
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• 2014

Objectives The purpose of this study was to investigate the effects of Cheunggihwadamhwan (CGHDH) extract on lowering lipid, antioxidation and production of proinflammatory cytokines in rats fed on high fat diet. Methods 40 Male Sprague-Dawley rats were fed on high fat diet for 8 weeks and 32 rats (above 400 g) were randomly divided into 4 groups (8 mice in each group) : control group, 100 mg/Kg CGHDH group, 200 mg/Kg CGHDH group, 300 mg/Kg CGHDH group. We fed a control group of rats a basal diet and administered normal saline(100 mg/kg, 1 time/1 day) for 4 weeks. And We fed each experimental group of rats basal diet and administered an extract of Cheunggihwadamhwan extracts (100 mg/kg, 200 mg/kg, 300 mg/kg, 1 time/1 day) for 4 weeks. At the end of the experiment, the rats were sacrificed to determine their chemical composition. We measured lipid in plasma and liver, concentration of proinflammatory cytokines, antioxidative activity and gene expression. The gene expression level was investigated by the way of reverse transcription-polymerase chain reaction (RT-PCR). Results 1. Concentration of plasma FFA, plasma TG, plasma total cholesterol and plasma LDL-cholesterol showed a significant decrement in Cheunggihwadamhwan groups. However, concentration of plasma HDL-cholesterol showed a significant increment in 200, 300 mg/kg Cheunggihwadamhwan groups. 2. Concentration of liver total cholesterol and liver TG showed a significant decrement in Cheunggihwadamhwan groups. 3. Concentration of plasma TBARS showed a significant decrement in all Cheunggihwadamhwan groups. Concentration of liver TBARS showed a significant decrement in 200, 300 mg/kg Cheunggihwadamhwan groups. Concentration of liver GSH-Px, SOD and CAT showed a tendency to decrease in all Cheunggihwadamhwan groups. 4. Concentration of plasma IL-$1{\beta}$, plasma IL-6, TNF-$\alpha$ and NO, showed a tendency to decrease in all Cheunggihwadamhwan groups. Concentration of plasma IL-10 showed a tendency to increase in all Cheunggihwadamhwan groups. 5. In the analysis of reverse transcription-polymerase chain reaction (RT-PCR), the gene expression of Apo-B and Apo-E in the Cheunggihwadamhwan groups showed a low expression than that of control group. The ratio of Apo-B expression per $\beta$-actin expression in the showed a significant decrement in all Cheunggihwadamhwan groups. The ratio of Apo-E expression per $\beta$-actin expression in the showed a significant decrement in 300 mg/kg Cheunggihwadamhwan groups. Conclusions According to this study, the extract of Cheunggihwadamhwan showed a positive effect of lowering lipid, antioxidation and a control of producing proinflammatory cytokines.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.18
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• pp.7825-7829
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• 2014

Objective: To investigate the effect of high expression of XAF1 in vivo or in vitro on lung cancer cell growth and apoptosis. Methods: 1. The A549 human lung cancer cell line was transfected with Ad5/F35 - XAF1, or Ad5/F35 - Null at the same multiplicity of infection (MOI); (hereinafter referred to as transient transfected cell strain); XAF1 gene mRNA and protein expression was detected by reverse transcription polymerase chain reaction (RT-PCR) and Western blotting respectively. 2. Methyl thiazolyl tetrazolium (MTT) and annexin V-FITC/PI double staining were used to detect cell proliferation and apoptosis before and after infection of Ad5/F35 - XAF1 with Western blotting for apoptosis related proteins, caspase 3, caspase - 8 and PARP. 3. After the XAF1 gene was transfected into lung cancer A549 cells by lentiviral vectors, and selected by screening with Blasticidin, reverse transcription polymerase chain reaction (RT-PCR) and Western blotting were applied to detect mRNA and protein expression, to establish a line with a stable high expression of XAF1 (hereinafter referred to as stable expression cell strain). Twenty nude mice were randomly divided into groups A and B, 10 in each group: A549/XAF1 stable expression cell strain was subcutaneously injected in group A, and A549/Ctrl stable cell line stable expression cell strain in group B (control group), to observe transplanted tumor growth in nude mice. Results: The mRNA and protein expression of XAF1 in A549 cells transfected by Ad5/F35 - XAF1 was significantly higher than in the control group. XAF1 mediated by adenovirus vector demonstrated a dose dependent inhibition of lung cancer cell proliferation and induction of apoptosis. This was accompanied by cleavage of caspase -3, -8, -9 and PARP, suggesting activation of intrinsic or extrinsic apoptotic pathways. A cell strain of lung cancer highly expressing XAF1 was established, and this demonstrated delayed tumor growth after transplantation in vivo. Conclusion: Adenovirus mediated XAF1 gene expression could inhibit proliferation and induce apoptosis in lung cancer cells in vitro; highly stable expression of XAF1 could also significantly inhibit the growth of transplanted tumors in nude mouse, with no obvious adverse reactions observed. Therefore, the XAF1 gene could become a new target for lung cancer treatment.