• 한국수산과학회지
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• 제30권6호
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• pp.984-991
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• 1997

New tumor suppressor gene, snm23, homologous to human nm23/NDP kinase (human nucleoside diphosphate kinase) gene whose product has a tumor metastasis inhibitory activity, was first cloned from Korean tiger shark (Scyliorhinus forazame) skin cDNA library constructed by using a $\lambda$ ZAP-II cDNA synthesis kit. About $1\times10^5$ plaques were screened and several positive plaques were isolated and confirmed by second screening. The phagemid containing a positive clone from the Uni-Zap XR vector was excised in vivo and the gene containing the tumor metastasis suppressor protein was named as snm23. Cloned gene, snm23, was sequenced with ABI-PRISM 310 Genetic Analyzer. The nucleotide and deduced amino acid sequences of snm23 have shown an open reading frame consisting of 450 base pairs that correspond to a protein of 150 amino acid residues, with a calculated molecular mass of 16.8 kDa. Sequence comparison of snm23 with human nm23/NDP kinase was performed by using Blast protein data base of National Center for Biotechnology Information. In order to determine tissue specificity, reverse transcription-polymerase chain reaction (RT-PCR) was used. Good expression level of snm23/NDP kinase was detected at the tissues from skin, cartilage, and liver of Korean tiger shark.

• Journal of Periodontal and Implant Science
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• 제44권3호
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• pp.141-146
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• 2014

Purpose: Peri-implantitis and periodontitis are inflammatory and infectious diseases of implant and tooth-supporting tissues. Recently, the role of gene polymorphisms of immune response components in the relevant pathogenesis has been investigated. The present study was the first to evaluate the relationship between two known single nucleotide polymorphisms (SNPs) of the receptor activator of nuclear factor kappa-${\beta}$ (RANK) gene (rs3018362 and rs35211496) in chronic periodontitis and peri-implantitis patients in an Iranian population. Methods: Eighty-one periodontally healthy patients, 38 patients with peri-implantitis, and 74 patients with chronic periodontitis were enrolled in this study. DNA was extracted from blood arm vein samples by using Miller's salting out technique according to the manufacturer's instructions given in the extraction kit. The concentration of DNA samples was measured using a spectrophotometer. The genetic polymorphisms of the RANK gene were evaluated using a competitive allele specific polymerase chain reaction (KBioscience allele specific PCR) technique. Differences in the frequencies of genotypes and alleles in the diseased and healthy groups were analyzed using chi-squared statistical tests (P<0.05). Results: Analysis of rs35211496 revealed statistically significant differences in the expression of the TT, TC, and CC genotypes among the three groups (P=0.00). No statistically significant difference was detected in this respect between the control group and the chronic periodontitis group. The expression of the GG, GA, and AA genotypes and allele frequencies (rs3018362) showed no statistically significant difference among the three groups (P=0.21). Conclusions: The results of this study indicate that the CC genotype of the rs35211496 RANK gene polymorphism was significantly associated with peri-implantitis and may be considered a genetic determinant for peri-implantitis, but this needs to be confirmed by further studies in other populations.

• Journal of Periodontal and Implant Science
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• 제34권1호
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• pp.205-221
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• 2004

Periodontal ligament(PDL) cells have been known as playing an important roles in periodontal regeneration and gingival fibroblasts are also important to periodontal regeneration by forming connective tissue attachment. There were rare studies about the gene expression patterns of PDL cells and gingival fibroblasts, therefore in this study, we tried cDNA microarray-based gene expression monitoring to explain the functional differences of PDL cells and gingival fibroblasts in vivo and to confirm the characteristics of PDL cells. Total RNA were extracted from PDL cells and gingival fibroblasts of same person and same passages, and mRNA were isolated from the total RNA using Oligotex mRNA midi kit(Qiagen) and then fluorescent cDNA probe were prepared. And microarray hybridization were performed. The gene expression patterns of PDL cells and gingival fibroblasts were quite different. About 400 genes were expressed more highly in the PDL cells than gingival fibroblasts and about 300 genes were more highly expressed in the gingival fibroblasts than PDL cells. Compared growth factor- and growth factor receptor-related gene expression patterns of PDL cells with gingival fibroblasts, IGF-2, IGF-2 associated protein, nerve growth factor, placental bone morphogenic protein, neuron-specific growth- associated protein, FGF receptor, EGF receptor-related gene and PDGF receptor were more highly expressed in the PDL cells than gingival fibroblasts. The results of collagen gene expression patterns showed that collagen type I, type III, type VI and type VII were more highly expressed in the PDL cells than gingival fibroblasts, and in the gingival fibroblasts collagen type V, XII were more highly expressed than PDL cells. The results of osteoblast-related gene expression patterns showed that osteoblast specific cysteine-rich protein were more highly expressed in the PDL cells than gingival fibroblasts. The results of cytoskeletal proteins gene expression patterns showed that a-smooth muscle actin, actin binding protein, smooth muscle myosin heavy chain homolog and myosin light chain were more highly expressed in the PDL cells than gingival fibrobalsts, and ${\beta}-actin$, actin-capping protein(${\beta}$ subunit), actin- related protein Arp3(ARP) and myosin class I(myh-1c) were more highly expressed in the gingival fibroblasts than PDL cells. Osteoprotegerin/osteoclastogenesis inhibitory factor(OPG/OCIF) was more highly expressed in the PDL cells than gingival fibroblasts. According to the results of this study, PDL cells and gingival fibroblasts were quite different gene expression patterns though they are the fibroblast which have similar shape. Therefore PDL cells & gingival fibroblasts are heterogeneous populations which represent distinct characteristics. If more studies about genes that were differently expressed in each PDL cells & gingival fibroblasts would be performed in the future, it would be expected that the characteristics of PDL cells would be more clear.

• 대한한방내과학회지
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• 제31권2호
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• pp.314-330
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• 2010

Objective : This study was performed to investigate the anti-fibrogenic effect and changes of inflammation-related genes by YBR I and YBR II (YBR I: Arteisiae Capillaris Herba, Atractylodis Rhizoma Alba, Hoelen/ YBR II: YBR I +Sanguisorbae Radix, Biotae Cacumen, Cirsii Japonici Herba) on HSC(hepatic stellate cells)-T6 and TAA-induced rat liver tissue. Materials and Methods : HSC-T6 were treated with various concentrations of distilled-water extract YBR I and YBR II extract for 24, 48 and 72 hours. After the treatment, cell viability, proliferation, procollagen levels and IL-6 levels were measured by using MTT Assay, BrdU Assay, Procollagen Type 1 C-peptide EIA kit, and Murine IL-6 ELISA Development kit. Rat liver fibrosis was induced by intraperitoneal TAA injection of 150mg/kg 3 times a week for 6 weeks. After the treatment, body weight, liver & spleen weights, liver function test, complete blood cell count and change of portal pressure were studied. In addition, gene expressions of ASMA, IL-6, MMP-2, TIMP-1 and TIMP-2, all of which are known to be associated with liver fibrosis, were analyzed by using Real-Time PCR. After YBR I and YBR IItreatment, percentages of collagen in TAA-induced rat liver tissue were measured. Results : The viability and proliferation of the HSC-T6 decreased as the concentration increased. The production of procollagen decreased as the concentration increased. The production of IL-6 was little influenced by YBR I and YBR II. There was no difference in rat body weight between the TAA-only group and the YBR groups. Compared with rat liver weight of TAA-only group, that of the YBR groups increased. In the YBR I group, the serum level of AST elevated by TAA injection significantly decreased and in the YBR I and II group, the serum level of ALP and ALT elevated by TAA injection decreased. In the YBR I group, white blood cell count elevated by TAA injection decreased but platelets increased. In the YBR I group, the portal pressure elevated by TAA injection significantly decreased. Decreases in the gene expression of ASMA and MMP-2 were observed in the YBR I group. The gene expression of IL-6 was little influenced by YBR I and YBR II -treated groups. In the histological finding, TAA injections caused severe fibrosis, but YBR I and YBR II treatment significantly reduced the amounts of hepatic collagens. Conclusions : These results suggest that YBR I and II have inhibitory effects on the hepatic fibrogenesis.

• Journal of Animal Science and Technology
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• 제49권5호
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• pp.559-568
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• 2007

최근 들어 유전자 또는 DNA 분절의 반복수 변이 (CNV)에 관한 연구가 다수 수행되었으며, 그 분석 방법 및 기기 또한 다양하게 발달되었다. 본 연구는 Landrace 품종의 KIT 유전자 CNV 탐지를 위하여 다른 분석 방법들에 비하여 정확도가 높은 정량적 OLA 기법(qOLA)을 이용하였다. qOLA와 pyrosequencing assay의 조합하여 분석한 결과를 Landrace 44두에 대한 좌표 분석을 한 결과 I1/I1 또는 I3/i(IBe), I1/I2 또는 I3/IP, I1/I3, I1/IP 또는 I2/i(IBe), I2/I2, I2/IP의 6 genotype으로 분류되었으며, PROC FASTCLUS procedure을 이용한 통계 분석과 좌표 분석을 상호 비교한 결과 genotype의 분류가 100% 일치하였다. 기기간의 차이점을 조사하기 위해 qOLA_3100과 qOLA _3130의 관측치 비교를 실시한 결과 동일한 genotype 분류결과를 얻었다. 또한 정밀성 및 정확도 비교에서 qOLA_3100의 경우 표준편차와 변이계수의 평균이 2.33과 4.10로 qOLA_3130(2.67과 4.81)에 비하여 낮게 나타났다. qOLA 반응전 PCR 산물에 대하여 proteinase K 처리효과를 분석한 결과 pro- teinase K를 처리한 경우 전기영동 분석시 noise peak들이 제거되었으며 각 genotype의 이론적 비율에 보다 정확히 일치하였다.

• 대한본초학회지
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• 제21권2호
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• pp.9-13
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• 2006

Objectives : To determine the DNA sequence of the 18S rRNA gene of the Rehmannia glutinosa and analyze it phylogenetically Methods : Dried root of the Rehmannia glutinosa was ground with a mortar and pestle. Glass beads(0.5 mm in diameter), TE buffer and SDS solution were added to that. The mixture was vortexed vigorously and extracted with the mixture of phenol, chloroform and isoamyl alcohol and with the mixture of the chloroform and isoamyl alcohol. The nucleic acids were precipitated with ethanol and resuspended in TE buffer. Contaminating RNA was digested with RNAse A and the DNA was purified further with the Geneclean Turbo Kit. This DNA was used as a template for amplification of the 18S rRNA gene by PCR. The PCR product was cloned in the pBluescript SK II plasmid by blunt-end ligation and the DNA sequence of the insert was determined. This DNA sequence was analyzed phylogenetically by the BLAST program. Results and Conclusion : Vortexing the ground powder of the dried plant root with glass beads during cell lysis improved recovery of DNA. The DNA sequence of the Rehmannia glutinosa 18S rRNA gene was determined and deposited at the GenBank as the accession number DQ469606. Phylogenetic analysis of that sequence showed the relationship between the members of the family of Scrophulariaceae and also the close relationship of the Buddleja davidii to the members of the Scrophulariaceae family.

• Asian Pacific Journal of Cancer Prevention
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• 제17권8호
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• pp.4149-4154
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• 2016

Background: Cervical carcinoma is one of the main causes of mortality in women worldwide as well as in India. It occurs as a result of various molecular events that develop from the combined influences of an individual's genetic predisposition and external agents such as smoking and menstrual hygiene, for example. However, infection with human papillomavirus (HPV) is the established major risk factor. The aim of the current study was to investigate p16 CpG island methylation and establish any correlation with mRNA expression in north Indian population. Materials and Methods: We analyzed 196 woman volunteer out of which 98 were cases and 98 healthy controls. For the analysis of methylation pattern, DNA extracted from blood samples was modified with a bisulfate kit and used as template for methylation specific PCR (MSP). Quantitative real-time PCR (QRT-PCR) was performed to check mRNA expression. Results: Correlation between methylation status of p16 gene and poor menstrual hygiene was significant (p=0.006), high parity cases showed methylation of p16 gene (p=0.031) with increased risk up to 1.86 times for cervical cancer and smoking was a strong risk factor associated with cervical cancer. We analyzed methylation pattern and found 60.3% methylation in cases with low mRNA expression level (0.014) as compare to controls (1.24). It was also observed that promoter methylation of p16 gene was significantly greater in FIGO stage III. Conclusions: We conclude that p16 methylation plays an important role in cervical cancer in the north Indian population and its methylation decreases mRNA expression. It can be used as an important and consistent blood biomarker in cervical cancer patients.

• The Korean Journal of Pain
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• 제19권2호
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• pp.168-174
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• 2006

Background: Several biochemical mediators, such as substance P, calcitonin gene-related peptide (CGRP) and prostaglandin $E_2$, have been demonstrated to be involved in herniated or degenerated disc-induced radiculopathy. The authors tested the hypothesis that these mediators would existed in the epidural space of humans. Methods: Thirty nine patients were divided into two groups; 27 patients, who were diagnosed with spinal stenosis (stenosis group), and 12 scheduled for epidural anesthesia, without a history of back pain (control group). Under fluoroscopic guidance, an epidural catheter was introduced through the caudal space and placed into the anterior and posterior spaces, up to and around the epidural adhesive area, in the stenosis group. In the control group, the catheter was placed into the posterior epidural space through the L3⁣-4 or L4⁣-5 intervertebral space. Epidural irrigation was performed with 10 ml of saline, via an epidural catheter. Aspirated lavage fluid was collected, and the concentrations of biochemical mediators (substance P, CGRP and prostaglandin $E_2$) measured using an enzyme immunoassay kit. Results: Substance P, CGRP and prostaglandin $E_2$ were detected in all the epidural lavage fluids from both groups. The concentrations of substance P and prostaglandin $E_2$ in the stenosis group were higher than those of the control (P < 0.05). However, there was no difference in the CGRP levels between the two groups. In the stenosis group, the concentrations of these three mediators in the anterior epidural space were no different to those in the posterior space. Conclusions: These results suggest that biochemical mediators, such as substance P and prostaglandin $E_2$, in the epidural space might be partly involved in pain mechanism associated with spinal stenosis.

• 한국동물생명공학회지
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• 제35권4호
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• pp.289-296
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• 2020

Spermatogonial stem cells are self-renewal and differentiate into sperm in post-pubertal mammals. There exists a balance between the self-renewal and differentiation in the testes. Spermatogonial stem cells make up only 0.03% of testicular cells in adult mice. These cells maintain sperm production by differentiating after puberty. Therefore, analyzing the expression of genes associated with spermatogenesis is critical for understanding differentiation. The present study aimed to establish the postnatal period of cells in relation to spermatogenesis. To study the expression of differentiated and undifferentiated marker genes in enriched spermatogonial stem cells, in vitro culture was performed and cells from pup (6-8-day-old) and adult (4-months-old) testicular tissues were isolated. As a result, undifferentiated genes, Pax7, Plzf, GFRa1, Etv5 and Bcl6b, were highly increased in cultured spermaotogonial stem cells compared with pup and adult testicular cells. On the other hands, differentiated gene, c-kit was highly increased in adult testicular cells, Also Stra8 gene was highly increased in pup and adult testicular cells. This study provides a better understanding of spermatogenesis-associated gene expression during postnatal periods.

• Clinical and Experimental Pediatrics
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• 제46권8호
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• pp.795-802
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• 2003

목 적 : 지방조직에 존재하는 OB 유전자에서 전사된 호르몬인 leptin은 여러 가지 생리적 요인이나, 호르몬에 의해서 영향을 받는다. Leptin의 발현에 대한 호르몬에 대한 연구가 많은 동물 실험들을 상대로 시도되고 있으나 사람에서 OB 유전자와 leptin 분비를 조절하는 호르몬의 영향 및 상호작용에 대해서는 아직 명확히 밝혀져 있지 않다. 본 연구는 사람의 복강에서 추출한 조직배양에서 OB 유전자와 leptin 분비를 조절하는 호르몬의 영향 및 상호작용에 대해서 알아보고자 하였다. 방 법 : 복부수술을 위하여 입원한 환자 7명을 대상으로 복강 내 지방조직을 절제하여 배양액에 호르몬을 첨가하지 않은 상태와 배양액에 인슐린, dexamethasone 및, 성장호르몬을 단독으로 첨가하거나, 인슐린과 dexamethasone을 동시에 첨가하거나, 인슐린과 dexamethasone과 성장호르몬을 같이 첨가한 상태에서 48시간 배양한 후 RNA를 추출하여 경쟁적 역전사 중합반응(competitive RT-PCR)을 시행하여 OB 유전자의 발현을 측정하고, human leptin IRMA Kit를 사용하여 지방조직에서 분비되는 배양액 내 leptin 양을 측정하였다. 결 과 : 인슐린은 단독으로는 OB 유전자 발현과 leptin 분비에는 영향을 미치지 못하였다. Dexamethasone은 OB 유전자의 발현과 leptin 분비를 증가시켰는데, 48시간 배양 후에 대조군에 비해 의미있게 증가하였다. 인슐린과 dexamethasone을 같이 배양시에는 OB 유전자 발현에 있어서는 의미있는 차이는 없었으나, leptin 분비는 48시간 배양 후 대조군에 비해 의미있게 증가하였다. 또한 성장호르몬 단독으로는 OB 유전자의 발현에 영향을 미치지는 못하나, 인슐린, dexamethasone, 성장호르몬을 같이 배양시에 인슐린과 dexamethasone의 OB 유전자 발현과 leptin 분비증가 능력을 억제시켰다. 결 론 : 인슐린 단독으로는 leptin 분비를 증가시키지 못하나, dexamethasone에 의해 상승작용이 나타나고, 이는 dexamethasone이 OB 유전자 발현을 증가시킨 후에 인슐린이 세포질내에서의 leptin 분비를 증가시킨다고 추정할 수 있다. 성장호르몬의 억제효과는 성장호르몬이 인슐린이나 dexamethasone에 대한 지방조직의 반응성을 변화시킴으로써 간접적으로 leptin의 발현을 조절할 것으로 추정되며, dexamethasone이 OB 유전자 발현을 증가시킨 후에 인슐린이 세포질 내에서의 leptin 분비를 증가시킨다는 것에 대한 연구가 더 필요하리라 사료된다.