• Title/Summary/Keyword: G-S PCR

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Genetic Diversity of Rehmannia glutinosa Genotypes Assessed by Molecular Markers (분자표지자에 의한 지황 유전집단의 유전적 다양성)

  • Bang, Kyong-Hwan;Chung, Jong-Wook;Kim, Young-Chang;Lee, Jei-Wan;Kim, Hong-Sig;Kim, Dong-Hwi
    • Journal of Life Science
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    • v.18 no.4
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    • pp.435-440
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    • 2008
  • Random amplified polymorphic DNA (RAPD) markers were used to identify the genetic diversities among and within varieties and landraces of Rehmannia glutinosa. Polymorphic and reproducible bands were produced by 10 primers out of total 20 primers used in the experiment. In RAPD analysis of the 11 genotypes, 64 fragments out of 73 amplified genomic DNA fragments were polymorphic which represented an average 6.4 polymorphic fragments per primer. Number of amplified fragments with random primers ranged from 2 (OPA-1) to 13 (OPA-11) and varied in size from 200 bp to 1,400 bp. Especially, OPA-10, OPA-11 and OPA-19 primers showed specific bands for varieties of Korea Jiwhang and Jiwhang il ho, which could be useful for discriminating from other varieties and landraces of R. glutinosa. Percentage polymorphism ranged from a minimum of 50% (OPA-1) to a maximum of 100% (OPA-11), with an average of 87.7%. Similarity coefficients were higher in the genotypes of Korea Jiwhang and Jiwhang il ho than in other populations. In cluster analysis, genotypes of Korea Jiwhang, Jiwhang il ho, and Japanese accession were separated from those of other varieties and landraces. Average of genetic diversity within the population $(H_S)$ was 0.110, while average of total genetic diversity $(H_T)$ was 0.229. Across all RAPD makers the $G_{ST}$ value was 0.517, indicating that about 52% of the total genetic variation could be explained by RAPDs differences while the remaining 48% might be attributable to differences among samples. Consequently, RAPD analysis was useful method to discriminate different populations such as domestic varieties and other landraces. The results of the present study will be used to understand the population and evolutionary genetics of R. gllutinosa.

Microbiological Quality Evaluation for Application of the HACCP System to the Bakery Products at Small Scale Bakeries (소규모 베이커리에서의 HACCP적용을 위한 미생물학적 위해도 평가)

  • 엄애선;권성희;정덕화;오상석;이헌옥
    • Korean journal of food and cookery science
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    • v.19 no.4
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    • pp.454-462
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    • 2003
  • Recently, the HACCP(Hazard Analysis Critical Control Point) system has been emphasized in food processing industries throughout the world. However, the system has, as yet, not been applied very well to domestic food industries. Due to the increase in the consumption of bakery products, more studies are required on the application of HACCP to establish the system in small-scale bakeries. This study was designed to provide basic data for setting management standards for HACCP, based on microbiological hazard evaluations of bakery products. Red bean paste filled breads, custard cream filled breads and cakes covered with fresh whipping cream were collected, and microbiological evaluations conducted on the raw materials, the manufacturing processes and potential hazards. The result showed the presence of coliforms in fresh cream of cakes and the soybean paste of soybean paste breads at levels as high as 105 CFU/g on the hazard analysis of the raw materials. Moreover, the general levels of bacteria and coliforms were over those of the standards during the intermediate fermentation and molding processes. Furthermore, high levels of coliforms were detected on the hands of the salesmen and bakers themselves. This suggests that the CCPs (Critical Control Points), such as fresh cream and red bean paste manufacturing process desperately require better management. There is also a requirement for education relating to personal hygiene for the production of hygienic bakery products and for the publics health.

Prevalence and Antimicrobial Resistance of Enterococus faecalis and Enterococcus faecium Isolated from Beef, Pork, Chicken and Sashimi (시판 축산물 및 수산물에서 Enterococcus faecalis와 Enterococcus faecium 분포 및 항생제 감수성에 관한 연구)

  • Sung, Chang-Hyun;Chon, Jung-Whan;Kwak, Hyo-Sun;Kim, Hyunsook;Seo, Kun-Ho
    • Food Science of Animal Resources
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    • v.33 no.1
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    • pp.133-138
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    • 2013
  • In this study, a total of 256 samples of retail raw meats (beef, pork and chicken) and sashimi were investigated for the presence of Enterococcus faecalis and Enterococcus faecium. We isolated a total of 117 E. faecalis and E. faecium from the samples, with contamination rates ranging from 18.8% for sashimi samples to 68.8% of chicken samples. E. faecalis was the predominant species recovered from all of the retail raw meats beef (42.2%), pork (42.2%), chicken (65.6%) and sashimi (12.5%). Among 117 isolates, 61 isolates (52.1%) were resistant to tetracycline, 32 isolates (27.4%) were resistant to erythromycin, 23 isolates (19.7%) were resistant to chloramphenicol, 16 isolates (13.7%) were resistant to ripampin, 10 isolates (8.5%) were resistant to gentamycin, 9 isolates (7.7%) were resistant to ciprofloxacin and 1 isolate (0.9%) was resistant to ampicillin and penicillin G. No resistance to amoxicillin + clavulanic acid and vancomycin was observed. Although no strain was resistant to vancomycin, the vanB gene was observed in 9 of 117 of Enterococcus (7.7%) demonstrating potential risk of vancomycin-resistant Enterococcus (VRE). Our results indicate that E. faecalis and E. faecium were highly prevalent in retail raw meats, but most strains were sensitive to tested antibiotics.

Is Genotypic Resistance-guide Eradication Therapy Effective for Patients with Refractory Helicobacter pylori Infection? (불응성 Helicobacter pylori 감염 환자들에게 유전자형 내성을 기반한 제균 치료는 유용한가?)

  • Kim, Sung Eun
    • The Korean journal of helicobacter and upper gastrointestinal research
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    • v.18 no.4
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    • pp.277-279
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    • 2018
  • 전 세계적으로 Helicobacter pylori의 항생제 내성률은 지속적으로 증가하고 있으며, 기존의 제균 치료에 실패한 H. pylori 감염 환자들에 대한 효과적인 구제요법(rescue therapy)의 필요성 역시 증가하고 있다. 이 연구는 두 개 기관, 공개, 평행 그룹, 무작위 배정 연구로서 불응성 H. pylori 감염 환자들의 구제요법으로 유전자형 내성을 기반한 치료(genotype resistance-guided therapy)와 경험적 치료(empirical therapy) 중 어느 것이 보다 효과적인지를 비교하고자 하였다. 2012년 10월부터 2017년 9월까지 20세 이상의 불응성 H. pylori 감염 환자들을 대상으로 하였으며, 불응성 H. pylori 감염은 과거 두 종류 이상의 H. pylori 제균 치료를 받았음에도 불구하고 H. pylori 제균에 실패한 환자들로 정의하였다. 이들에게서 한 군은 14일간의 유전자형 내성을 기반한 순차 치료(n=21 in trial 1, n=205 in trial 2)를, 다른 한 군은 환자들의 과거 제균 치료 종류를 감안한 14일간의 경험적 순차 치료(n=20 in trial 1, n=205 in trial 2)를 시행하였다. 순차 치료법은 첫 7일은 esomeprazole 40 mg과 amoxicillin 1 g을 하루 두 번 복용한 다음, 나머지 7일은 esomeprazole 40 mg과 metronidazole 500 mg, 그리고 1) levofloxacin 250 mg 또는 2) clarithromycin 500 mg 또는 3) tetracycline 500 mg을 하루 두 번 복용하는 것으로 구성하였다. 23S ribosomal RNA (rRNA)나 gyrase A에 대한 내성 관련 돌연변이 여부는 direct sequencing을 통한 중합효소연쇄반응(polymerase chain reaction, PCR) 검사를 이용하였고, 제균 성공 여부는 요소호기검사를 통해 확인하였다. 일차 결과 지표는 치료 방법에 따른 제균율로 정하였다. Trial 1에서는 tetracycline 대신 doxycycline 100 mg을 사용하였는데, 제균 성공률이 유전자형 내성을 기반한 치료군에서는 17명(81%), 경험적 치료군에서는 12명(60%)으로 나타났다(P=0.181). 하지만, 다른 순차 치료군들과 비교하였을 때, doxycycline을 포함한 순차 치료군의 제균율이 현저히 낮은 것으로 나타나서(15/26, 57.7%) doxycycline을 포함한 순차 치료법은 종결하기로 하고, trial 2부터는 doxycycline 대신 tetracycline으로 교체하여 연구를 지속하였다. Trial 2의 intention-to-treat (ITT) 분석 결과, 유전자형 내성을 기반한 치료군에서는 160/205명(78%), 경험적 치료군에서는 148/205명(72.2%)으로 두 그룹 간의 통계적인 제균율의 차이는 보여주지 못하였다(P=0.170). 부작용 및 환자 순응도에서도 양 군 간의 의미 있는 차이는 없었다. 따라서, 두 종류 이상 H. pylori 제균 치료에 실패한 환자들이라고 할지라도 기존의 제균 치료력을 바탕으로 적절한 경험적 치료를 시행하는 것은 유전자형 내성을 기반한 치료 정도의 효과는 있으며 접근성, 비용, 환자들의 선호도 등의 여러 가지 부가적인 사항들을 고려할 때, 제균 치료력을 고려한 경험적 치료는 간단한 수준의 유전자형 내성을 기반한 치료의 대안으로 받아들여질 수 있을 것으로 제안하였다.

Effects of Coptidis Rhizoma on the Anti-inflammation and Motor Recovery in Photothrombotic Brain Infarction Model in Rats (광화학적 뇌경색 백서 모델에서 황련의 항염증 및 운동기능 회복에 미치는 효과)

  • Lee, Su-Kyung;Lee, In;Shin, Sun-Ho;Kim, Eun-Young;Shin, Byung-Cheul
    • The Korea Journal of Herbology
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    • v.24 no.1
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    • pp.179-189
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    • 2009
  • Objectives : Coptidis Rhizoma (Coptis japonica MAKINO; CR) is a well known crude drug as antimicrobial, antibacterial, anti-inflammatory, antioxidant activity. However, there is no study of the effect of CR on brain infarction and it's mechanism. The aim of this study was to investigate the effects on ischemic stroke induced by photothrombotic infarction by evaluating the functional & neuronal recovery after brain infarction. Materials & Methods : Male Sprague-Dawley rats (250-300 g) were induced photothrombotic brain infarction on sensorimotor cortex, and brain infarction volume by image J software (NIH, USA) after Nissl stain, also single pellet reaching task as a functional motor recovery were observed. After orally pretreated by CR (500 mg/kg) or normal saline as a sham control before 7 days from the time of photothrombotic infarction, rats were sacrificed. After then we analysed anti-inflammatory cytokines (TNF-$\alpha$, IL-6, IL-1$\beta$), by RT-PCR and ELISA method, and immunohistochemistry (GFAP, connexin-43) as a marker of neural plasticity. Results : CR (100, 250, 500 mg/kg) decreased the infarction volume dose-dependently, however the effect of 500mg/kg of CR (CR 500) showed the best (P=0.051). Also, CR 500 decreased the infarction volume time-dependently, the most effective time was 3-7 days after stroke. Photothrombosis increased inflammatory cytokines after infarction, CR 500 suppressed significantly mRNA expression of IL-1$\beta$, IL-6 and TNF-$\alpha$. In serum, CR 500 decreased the amount of IL-1$\beta$, 12h, 24h and 48h respectively (p < 0.05), also decreased that of IL-6 and TNF-$\alpha$, 12h respectively (p < 0.05) after infarction. The more astrocytes were observed and neural plasticity was facilitated in the rat brain of CR 500 than that of sham control in immunohistochemistry. Conclusions : This results suggest that CR decrease infarction volume and improve functional motor recovery in acute stage in photothrombotic ischemic infarction model in the mechanism of anti-inflammation and promoting neural plasticity.

β-elemene Induces Caspase-dependent Apoptosis in Human Glioma Cells in vitro through the Upregulation of Bax and Fas/FasL and Downregulation of Bcl-2

  • Li, Chen-Long;Chang, Liang;Guo, Lin;Zhao, Dan;Liu, Hui-Bin;Wang, Qiu-Shi;Zhang, Ping;Du, Wen-Zhong;Liu, Xing;Zhang, Hai-Tao;Liu, Yang;Zhang, Yao;Xie, Jing-Hong;Ming, Jian-Guang;Cui, Yu-Qiong;Sun, Ying;Zhang, Zhi-Ren;Jiang, Chuan-Lu
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.23
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    • pp.10407-10412
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    • 2015
  • Background: ${\beta}$-elemene, extracted from herb medicine Curcuma wenyujin has potent anti-tumor effects in various cancer cell lines. However, the activity of ${\beta}$-elemene against glioma cells remains unclear. In the present study, we assessed effects of ${\beta}$-elemene on human glioma cells and explored the underlying mechanism. Materials and Methods: Human glioma U87 cells were used. Cell proliferation was determined with MTT assay and colony formation assay to detect the effect of ${\beta}$-elemene at different doses and times. Fluorescence microscopy was used to observe cell apoptosis with Hoechst 33258 staining and change of glioma apoptosis and cell cycling were analyzed by flow cytometry. Real-time quantitative PCR and Western-blotting assay were performed to investigated the influence of ${\beta}$-elemene on expression levels of Fas/FasL, caspase-3, Bcl-2 and Bax. The experiment was divided into two groups: the blank control group and ${\beta}$-elemne treatment group. Results: With increase in the concentration of ${\beta}$-elemene, cytotoxic effects were enhanced in the glioma cell line and the concentration of inhibited cell viability ($IC_{50}$) was $48.5{\mu}g/mL$ for 24h. ${\beta}$-elemene could induce cell cycle arrest in the G0/G1 phase. With Hoechst 33258 staining, apoptotic nuclear morphological changes were observed. Activation of caspase-3,-8 and -9 was increased and the pro-apoptotic factors Fas/FasL and Bax were upregulated, while the anti-apoptotic Bcl-2 was downregulated after treatment with ${\beta}$-elemene at both mRNA and protein levels. Furthermore, proliferation and colony formation by U87 cells were inhibited by ${\beta}$-elemene in a time and does-dependent manner. Conclusions: Our results indicate that ${\beta}$-elemene inhibits growth and induces apoptosis of human glioma cells in vitro. The induction of apoptosis appears to be related with the upregulation of Fas/FasL and Bax, activation of caspase-3,-8 and -9 and downregulation of Bcl-2, which then trigger major apoptotic cascades.

The Effect of Treponema denticola immunoinhibitory protein on cytokine expression in T cells (Treponema denticola 면역억제 단백질이 T 세포의 cytokine 발현에 미치는 영향)

  • Lee Sang-Yup;Shon Won-Jun;Lee WooCheol;Baek Seung-Ho;Bae Kwang-Shik;Lim SungSam
    • Restorative Dentistry and Endodontics
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    • v.29 no.5
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    • pp.479-484
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    • 2004
  • Immunoinhibitory protein extracted from sonicated Treponema denticola have been shown to suppress cell cycle progression of human lymphocytes. To study in detail about the effect of this microorganism on the function of lymphocytes. we investigated the levels of Interleukin-2 (IL-2) and Interleukin-4 (IL-4) production by T lymphocytes before and after the addition of $12.5{\;}\mu\textrm{g}/ml$ T. denticola sonicated extracts. In this study. levels of IL-2 and IL-4 produced from T cells pretreated with sonicated extracts were evaluated by using the quantitative sandwich enzyme immunoassay technique. In response to phytohemagglutinin (PHA) stimulation. T cell produced increased levels of IL-2 and IL-4. However. the expressions of both cytokines were significantly inhibited when PHA activated-T cells were pre-exposed to sonicated T. denticola extracts (p < 0.05). These findings suggest that the T. denticola sonicated extracts induced-immunosuppression in Th1 and Th2 cell functions could be a part of the pathogenic mechanism of the endodontic failure associated with this microorganism.

Polymorphisms in Glutamate Receptor, Ionotropic, N-methyl-D-aspartate 2B(GRIN2B) Genes of Autism Spectrum Disorders in Korean Population : Family-based Association Study (한국인 자폐스펙트럼장애에서 Glutamate Receptor, Ionotropic, N-methyl-D-Aspartate 2B(GRIN2B) 유전자 다형성-가족기반연구)

  • Yoo, Hee Jeong;Cho, In Hee;Park, Mira;Yoo, Hanik K.;Kim, Jin Hee;Kim, Soon Ae
    • Korean Journal of Biological Psychiatry
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    • v.13 no.4
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    • pp.289-298
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    • 2006
  • Objectives : Autism is a complex neurodevelopmental spectrum disorder with a strong genetic component. Previous neurochemical and genetic studies suggested the possible involvement of glutamate N-methyl-D-aspartate(NMDA) receptor in autism. The aim of study was to investigate the association between the NMDA2B receptor gene(GRIN2B) and autism spectrum disorders(ASD) in the Korean population. Methods : The patients with ASD were diagnosed with Autism Diagnostic Interview-Revised and Autism Diagnostic Observation Schedule based on DSM-IV diagnostic classification. The present study was conducted with the detection of four single nucleotide polymorphisms(SNPs) in GRIK2 and family-based association analysis of the single nucleotide polymorphisms in Korean ASD trios using transmission disequilibrium test (TDT). Results : One hundred twenty six patients with ASD and their biological parents were analyzed. 86.5% were male and 85.1% were diagnosed as autistic disorder. The mean age was $71.9{\pm}31.6$ months(range : 26-185 months). We found that rs1805247 showed significantly preferential transmission(TDT ${\chi}^2$=12.8, p<0.001) in ASD. Conclusion : One SNP in GRIN2B gene was significantly associated with ASD in the Korean population. This result suggests the possible involvement of glutamate NMDA receptor gene in the development of ASD.

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Efficient Remediation of Petroleum Hydrocarbon-Contaminated Soils through Sequential Fenton Oxidation and Biological Treatment Processes (펜톤산화 및 생물학적 연속처리를 통한 유류오염토양의 효율적 처리)

  • Bae, Jae-Sang;Kim, Jong-Hyang;Choi, Jung-Hye;Ekpeghere, Kalu I.;Kim, Soo-Gon;Koh, Sung-Cheol
    • Korean Journal of Microbiology
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    • v.47 no.4
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    • pp.356-363
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    • 2011
  • The accidental releases of total petroleum hydrocarbons (TPH) due to oil spills frequently ended up with soil and ground water pollution. TPH may be degraded through physicochemical and biological processes in the environment but with relatively slow rates. In this study an attempt has been made to develop an integrated chemical and biological treatment technology in order to establish an efficient and environment-friendly restoration technology for the TPH contaminated soils. A Fenton-like reaction was employed as a preceding chemical treatment process and a bioaugmentation process utilizing a diesel fuel degrader consortium was subsequently applied as a biological treatment process. An efficient chemical removal of TPH from soils occurred when the surfactant OP-10S (0.05%) and oxidants ($FeSO_4$ 4%, and $H_2O_2$ 5%) were used. Bioaugmentation of the degrader consortium into the soil slurry led to an increase in their population density at least two orders of magnitude, indicating a good survival of the degradative populations in the contaminated soils ($10^8-10^9$ CFU/g slurry). TPH removal efficiencies for the Fenton-treated soils increased by at least 57% when the soils were subjected to bioaugmentation of the degradative consortium. However, relatively lower TPH treatment efficiencies (79-83%) have been observed in the soils treated with Fenton and the degraders as opposed to the control (95%) that was left with no treatment. This appeared to be due to the presence of free radicals and other oxidative products generated during the Fenton treatment which might inhibit their degradation activity. The findings in this study will contribute to development of efficient bioremediation treatment technologies for TPH-contaminated soils and sediments in the environment.

The effects of enamel matrix derivatives on the proliferation and gene expression of PDL fibroblast, $SaOs_2$ cells and Cementum derived cells (법랑기질유도체가 치주인대세포, 불멸화 조골세포, 백악질 유래세포의 증식과 유전자 발현에 미치는 영향)

  • Jeong, Yoo-Jee;Kim, Kyoung-Hwa;Kim, Tae-Il;Seol, Yang-Jo;Ku, Young;Lee, Hae-Jun;Rhyu, In-Chul;Chung, Chong-Pyoung;Han, Soo-Boo;Lee, Yong-Moo
    • Journal of Periodontal and Implant Science
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    • v.35 no.2
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    • pp.321-333
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    • 2005
  • 1. 목적 in vitro 상에서 법랑기질유도체가 치주인대섬유아세포, 불멸화 조골세포와 백악질 유래세포의 증식과 유전자 발현에 미치는 영향을 알아보고자 하였다. 2. 연구방법 및 재료 <세포증식 연구> 교정을 목적으로 발거한 치아에서 분리, 배양한 치주인대섬유아세포와 백악질유래세포, 그리고 $SaOs_2$ 세포를 이용하였다. 법랑기질유도체가 세포 증식에 미치는 영향을 알아보기 위해, 35 mm Petri dish에 dish 당 $5{\times}10^3$ 개의 세포를 접종하였다. 대조군은 1% 항생제와 10% FBS를 포함한 DMEM 배지를 이용했고, 5mM 초산을 첨가한 군과 첨가하지 않은 두 개의 대조군이 이용되었다. 실험군은 100 ${\mu}g/ml$의 법랑기질유도제를 첨가한 군과 100 ${\mu}g/ml$의 법랑기질유도체와 5 mM의 초산을 첨가한 2개의 실험군이 이용되었다. 각 군은 세 개의 배양접시에 행해졌고, 1, 3, 8일에 세포의 수를 각각 측정하였다. 결과는 repeated measures ANOVA로 통계 처리하였다. <유전자 발현 연구> 각 세포의 형질 특성을 알아보기 위해 RT-PCR을 실시하여 조골세포 분화 표식자와 연관된 Human collagen type I(COL I), human osteopontin(OP), human osteocalcin(OC), human alkaline phosphatase(ALP)와 human bone sialoprotein(BSP)의 mRNA 발현을 실험 1, 3, 8일에 걸쳐, 세 군의 차이를 비교 관찰하였다. 3. 결과 <세포증식 연구> 치주인대세포와 백악질유래세포, 그리고 $SaOs_2$ 세포의 증식은 법랑기질유도체에 의해 영향을 받지 않았다. 대조군과 초산이 포함된 대조군 그리고 법랑기질유도체와 초산이 포함된 실험군에서 유의할 만한 세포 수의 차이가 실험 기간 1, 3, 8일에 걸쳐 나타나지 않았다(p<0.05). <유전자 발현 연구> ALP와 COL I은 세 군의 세포에서 모두 발현되었고, 발현 정도는 EMD에 영향을 받지 않았다. OC은 세 군에서 모두 비교적 약하게 발현되었고, 특히 $SaOs_2$ cell과 백악질유래세포에서 약하게 발현되었다. EMD는 OC의 발현정도를 약하게 하였다. OP은 백악질유래세포에서 1, 3, 8일에 걸쳐 EMD 유무에 관련 없이 발현되지 않았다. 그러나 치주인대세포와 $SaOs_2$ cell에서는 강하게 발현되었다. BSP는 치주인대세포와 $SaOs_2$ cell에서 1, 3, 8일에 걸쳐 비교적 고르게 발현되었다. EMD 배지에서 배양된 백악질유래세포는 8일에는 BSP가 발현되지 않았다. 4 결론 이번 실험 결과에 의하면 법랑기질유도체는 치주인대세포, 불멸화 조골세포와 백악질 유래세포의 증식에 있어 유의성 있는 효과를 나타내지 않았다. 그러나, 유전자 발현에 있어서는, 치주인대세포와 백악질유래세포, 그리고 $SaOs_2$ 세포 모두에서 OC mRNA의 발현을 억제하는 효과를 나타내었다. EMD는 세포의 증식에는 영향을 미치지 않지만, 유전자 발현에 있어 일부 영향을 미치는 것으로 보인다. 법랑기질유도체가 세포의 증식과 유전자 발현에 미치는 영향은 배양된 세포의 형질특성, 배양환경, 배양일수 등에 따라 달라질 수 있다. 그러므로 법랑기질유도체가 in vitro 상에서 세포에 미치는 영향은 보다 정량화된 연구가 필요하다.