Slide Session : OS-IFD-07 ; Infectious Disease : In Vitro Antiviral Activity of Ribavirin Against Severe Fever with Thrombocytopenia Syndrome Virus

( Myung Jin Lee ),( Kye Hyung Kim ),( Jong Youn Yi ),( Su Jin Choi ),( Chung Jong Kim ),( Nak Hyun Kim ),( Kyoung Ho Song ),( Pyoeng Gyun Choi ),( Ji Hwan Bang ),( Wan Beom Park ),( Eu Suk Kim ),( San 대한내과학회 2014 대한내과학회 추계학술대회 Vol.2014 No.1

In Vitro Antiviral Activity of Ribavirin Against Severe Fever with Thrombocytopenia Syndrome Virus Myung Jin LEE1, Kye-Hyung KIM1, Jongyoun YI2, SuJin CHOI1, Chung-Jong KIM1, Nak- Hyun KIM1, Kyoung-Ho SONG1, Pyoeng Gyun CHOI1, Ji-Hwan BANG1, Wan Beom PARK1, Eu Suk KIM1, Sang-Won PARK1, Hong Bin KIM1, Nam Joong KIM1, Myoung- Don OH1 Seoul National University College of Medicine, Korea1, Pusan National University School of Medicine, Korea2 Background: Severe fever with thrombocytopenia syndrome (SFTS) is an emerging infectious disease caused by a novel Bunyavirus, severe fever with thrombocytopenia syndrome virus (SFTSV). No effective antiviral therapy is proven yet, but clinical use of ribavirin (RBV) has been tried. We investigated the antiviral effect of RBV against SFTSV in vitro. Methods: To test for cytotoxicity of RBV, Vero cells were treated with different concentrations of RBV (3.90 to 500 μg/mL, two-fold dilution) and analyzed by cell viability MTS assay 48h post-infection. To determine antiviral activity of RBV against SFTSV, Vero cells were infected with SFTSV strain Gangwon/Korea/2012 at 100 TCID50 (50% tissue culture infective dose) per well in a 96-well plate, and RBV was added at the concentrations showing no or minimal cytotoxicity. Viral RNAs were extracted from the culture supernatants and quantifi ed using one-step real-time reverse transcription- PCR to amplify the partial large segment of SFTSV. Statistical analysis was done by one-way ANOVA with Tukey`s post hoc test. Results: Cytotoxicity due to RBV was not observed at RBV concentration =31.3 μg/ mL. Viral RNAs at 24h post-RBV treatment were reduced with increasing RBV concentrations (1-32 μg/mL), compared with those of mock-treated cells (P <0.01, Figure). Half maximal inhibitory concentration (IC50) of RBV was 3.69 μg/mL at 24h post-RBV treatment. Conclusions: Our study shows that RBV has antiviral effect against SFTSV in a dose-dependent manner. Further studies are required to evaluate the effi cacy of RBV in SFTS.

뇌졸중 환자의 일상생활동작, 우울과 삶의 질의 관계

김원옥 ( Won Ock Kim ),강현숙 ( Hyun Sook Kang ),왕명자 ( Jmyung Ja Wang ),김정화 ( Jeong Hwa Kim ),최진이 ( Jin Yi Choi ) 경희대학교 동서의학연구소 2007 동서간호학연구지 Vol.13 No.2

본 연구는 뇌졸중 환자의 일상생활동작, 우울, 삶의 질의 관계를 파악하고자 하는 서술적 조사 연구로서 일상생활동작,’ 우울, 삶의 질 정효〉 관계가 있음을 확인하였다. 그러므로 본 안구의 주요한 결과를 중심으로 다음과 같이 고찰하였다. 뇌졸중 환자들의 일상활동 수행능력을 측정하는 도 구로 FIM, Modificd Barthel Index (MBI) 등이 많이 쓰 이고 있는데 FIM은 분야에 상관없이 표준화를 통한 높은 실용성,실행성, 이해도로· 인하여 널리 사. 하고 있는 도구로 이를 이용하여 측정한 본 연구에서는 872점으로 국내에서 만성 뇌졸중 환자와 퇴원 시 뇌졸중 환자를 대상으로 측정한 85. 9점 (Hw떼g, 2004) 과 85. 3점 (Kim, 2004) , 85. 8점 (Hong et al., 1998) 에 비하 여 약간 높아졌으나 비슷하였고, 일본 뇌졸중 환자를 대상으로 한 연구의 77. 1점보다는 높은 결과이었다 (Hama et al., 2007) . 특히 하부역역인 운동영역은 다 른 연구에 비해 높았으나 인지영역은 다른 연구에 비 해 낮은 편이었다 (Hwang, 2004二 Hama et al. , 2007, Hong 嫩 al. , 200긱 Kim, 2004) . 미국의 지역사회시설 에 있는 뇌졸중이 처음 발병한 환자들을 대상으로 재 활 후 5년간 환자를 추적한 연구에서 3년 후까지는 FIM점수가 상승하였으나 그 이후 감소되는 양상을 보였고 본 연구에서는 친단 후 경과 기간이 35.5개월 (87.2점)로 퇴원 후 3년 된 시점의 결과no도4점) 와 비교하였을 때 낮은 편이었다〈 Leonard, Mnler, Griffiths, Mcclatchie, & wherry, 1998) . 이러한 결과는 지역사 회와 집에서 간호를 받고 있는 뇌졸중 환자들에게 지속적인 재활 프로그램의 필요성이 높다는 것을 시사 한다고 볼 수 있다. 그러므로 입원을 한 직후부티 환 자에게 적절한 재활프로그램을 계획하여 시작하고 퇴원 후에는 연계되어 재활을 받을 수 있는 시설과 연결하여 환자가 지속적으로 재활을 할 수 있도록 돕는 병원과 지역사회가 공동으로 개발하고 시행하는 개활프로그램이 필요하다. 우울은 뇌졸중을 진단 받은 환자들의 급성기와 회 복기에 많이 동반되는 증상으로 본 연구에서도 CES- D를 이용하여 우울을 측정한 결과 23. 8점으로 Bak (2003) 의 연구와 비슷한 정도로 높게 측정되어 이 전의 연구결과와 같이 뇌졸중 환자의 가장 흔한 후유 증이 우울임을 다시 확인할 수 있었다 (Cho et al. 2003, `` Chon & Rhee, 1992, `` Kim, 2005, `` Kim, Seo, & Kang, 2000; singh et al. , 2000) . 그러므로 입원 시부터 신체적인 증상 뿐 아니라 우울도 같이 사정하고 우울이 진행되지 않도록 지속적인 관찰과 정서적인 간호가 뇌졸중 환자를 위한 재활프로그램에 포함이 되어야 한다. 삶의 질은 3 ``03점으로 뇌졸중 환자 l이명을 대상으로 한 chung (2000) 의 연구에서 의 3. 41점보다는 낮았으나 Kim과 Kim (2003) 의 연구의 3. 17점과는 비슷하였고 전체적으로 낮은 편이었다. 이런 결과는 최근 의학기술의 발전으로 뇌졸중으로 인한 사망률이 감소 되고 만성적인 뇌졸중 환자가 증가함에도 불구하고 재가뇌졸중 환자들에 대한 재활과 관리가 효과적으로 이루어지지 않고 있음으로 확인할 수 있었다. 따라서 뇌졸중과 삶의 질은 단기간에 향상되는 것이 아니므로 개활 프로그램이 단기적인 것보다는 체계적으로 계획되고 지속적으로 수행되어져 삶의 질을 관찰하는 연구가 필요할 것으로 생각된다. 또한 일상생활 동작을 잘 할수록 우울점수가 낮게 나타났고 삶의 질 정도가 높게 나타났다. 그리고 우 울 정도가 심할수록 삶의 질 정도가 낮은 것으로 나타났다. 뇌졸중 환자들에 대한 선행연구결과들을 살 펴보면 뇌졸중은 일상생활동작 수행과 같은 신체적인 문제, 우울, 대인관계 감소 등의 사회적 문제가 발생 되고 장기적인 치료와 간호를 요구하는 질병으로서 최종적으로. 삶의 질이 저하되는 것을 보고하고 있다 (Chung, 2000; Hama et al. , 2007, `` Ingrid, Kwakkel, Bruin, & Lindeman, 2007; Khan, 2004; Kim & Kim, 2003; singh et al. , 2007) . 이러한 연구 결과는 일상생활동작, 우울, 삶의 질이 단계적으로 순환되는 관계로 서 모두 뇌졸중환자를 위한 재활프로그램 내용에 포함되어야 함을 의미한다. 이런 연구결과들을 종합하여 볼 때 뇌졸중은 환자 의 뇌손상을 일으켜 장기적인 우울과 신체적인 기능 장애를 발생시킬 가능성이 높으므로 뇌졸중 환자의 재활프로그램은 입원을 하자마자 계획하여 치료와 동 시에 시작하고 특히 우울은 뇌졸중의 만성적인 건강 문제로 재활과정을 방해할 수 있으므로 신체적인 중 재와 더불어 정서적인 중재를 포함한 포괄적인 내용 의 프로그램이 바람직하고 장기적으로 삶의 질을 평 가함이 바랑직할 것이다. 또한 뇌졸중은 장기적인 재 활이 요구되는 질환이므로 단기간의 재활 프로그램을 병원에서만 시행하는 것이 아니라 퇴원 후 병원과 지 역 사회에서 연계하여 치료적인 연속성을 가지고 유지 할 수 있는 재활 프로그램의 개발이 반드시 필요할 것이 라고 본다. Introduction: The purpose of this study was to examine relationships among stroke patients`` activities of daily living(ADL), depression, and quality of life(QOL). Methods: Data were collected by questionnaires from 251 stroke patients at 5 hospitals and 2 public health centers in Seoul and Kyonggi-Do. The following Questionnaires were utilized for this study: Functional independence measure(FIM) from Grange, Hamilton, and Sherwin(1986), the Korean CES-D scale from Randloff(1977), and quality of life scale from Bang, Hwal-Lan(1991). Results: Data were analyzed using the pearson correlation, t-test, and ANOVA with SPSS/WIN 12.0. The results of this study were as follow; The subjects consisted of 110 females and 141 males. The mean score of activity of daily living was 87.2(range 18-126). Age, family history, comorbidity, paralysis site, and duration after diagnosis were significantly associated with activity of daily living. The mean score of depression was 23.8(range 11-44). Age, gender, education, and job were significantly associated with depression. The mean score of quality of life scale was 3.0(range 1-5). Gender, religion, education, job, economic status, and duration after diagnosis were significantly associated with quality of life. Relationships among activity daily living, depression, and quality of life scale: The activity of daily living had a negative relationship with depression score. The activity of daily living had a positive relationship with QOL score. The depression had a negative relationship with QOL score. Conclusion: Based on these results, it is necessary to run rehabilitation programs to improve stroke patients`` ADL, depression. Also, it research for of identifying stroke patients`` quality of life according to rehabilitation program.

항암제 처리한 백혈병 세포주에서의 Apoptosis 발현

김삼용,윤소현,김현수,김종숙,윤환중,김진경,조덕연 충남대학교 암연구소 1998 癌共同硏究所 硏究誌 Vol.2 No.1

The bcl-2 proto-oncogene encodes a 26 kD protein that promotes cell survival by blocking apoptosis. The bax protein is a member of the bcl-2 familly, now known to form heterodimers with the bcl-2 protein. The ratio of bax to bcl-2 is be critical in determining the fate of the cell in response to stimuli that can induce apoptosis. Extract of Pulsatilla Koreana (SB-31) showed promising antitumor activity in vitro with Topo I inhibitory action. In the present study, the relationship between apoptosis and the apoptosis related proteins, bcl-2 and bax were investigated in human leukemic cell lines HL-60, U-937 and CEM-CM3. All anticancer drugs(adriamycin, etoposide, camptothecin, SB-31) induced extensive apoptosis in HL-60, U-937 cells and CEM-CM3 cells. The expression of bcl-2 and bax protein were determined in cell lines by western blotting before and after incubation with anticancer drugs at different time points. 1) In HL-60 or U-937 cell lines, down regulation of bcl-2 and up-regulation of bax were found after incubation with ADR, VP-16 or camptothecin. 2) In HL-60 or U-937 cell lines, no significant change in bcl-2 or bax protein expression resulted after incubation with SB-31. 3) In CEM-CM3 cells, virtually no change was noted in bcl-2/bax expression after incubation with ADR, VP-16, camptothecin or SB-31. It is suggested that different leukemic cell lines use different pathways of apoptosis activation and a given cell may utilize different pathways of apoptosis activation in response to different cytotoxic agents.

자료포락분석(DEA)를 이용한 보건의료의 효율성 측정

김남송,박홍섭,김은진,김설화,김정숙,김현아,김경숙,신지혜,최은정,양은진,유선미,유수진 圓光大學校 醫科學硏究所 2007 圓光醫科學 Vol.22 No.1

정상과 갑상선 종양조직에서 사람 IGF-I 유전자의 발현

김성운,장현하,박상미,김덕윤,우정택,양인명,김진우,김영설,김광원,고석환,홍성화,최영길 경희대학교 유전공학연구소 1993 遺傳工學論文集 Vol.5 No.-

Many of the growth-promoting properties of growth hormone(GH) are mediated by insulin-like growth factor-I(IGF-I), a highly conserved circulating 70-amino acid peptide. Recent studies have shown that multiple mechanisms influence IGF-I gene expression, including transcription from two promoters, alternative RNA splicing, and variable polyadenylation. In thyroid tissue, thyroid stimulating hormone(TSH) and IGF-I are the most possible candidates for follicular cell proliferation and hypertrophy. Actually IGF-I had autocrine and paracrine effect for tissue growing. We prepared thyroid tumor tissue mRNAs using single step method for detecting IGF-I levels according to different tissues, i.e., thyroid adenoma or papillary thyroid carcinoma. We used Northern blot analysis for IGF-I mRNA and RNase protection assay (RPA) for IGF-I transcription start sites. For Northern blot, we used whole human IGF-I cDNA as a DNA probe and for RPA, we used IGF-I exon 1 containing noncoding promoter 1 as a riboprobe. We got good RNA bands from Northern blot analysis around 1 kb (IGF-IA) and 7.5 kb (IGF-IB) region. To clarify the amount of both IGF-IA and IB mRNAs, we measured autoradiographied signal of IGF-I mRNAs bands using densitometer. In IGF-IA signals, there's no change among liver and thyroid tissues, but in case of IGF-IB mRNA bands, the signal was markedly increased in thyroid carcinoma tissues than that of normal thyroid tissue (85% vs 14%). In the study of RPA, all thyroid tissues used the same transcription start sites as those of liver's. We concluded that that this different regulation of IGF-I mRNA was originated from tissue specificity. That meant some tissue specific transcription factor/s were related to tissue IGF-I expression.

조혈모세포이식 환자에서 침습성 진균 감염에 대한 Micafungin의 예방 효과 및 안전성

김시현,이동건,최수미,권재철,박선희,최정현,유진홍,이성은,조병식,김유진,이석,김희제,민창기,조석구,김동욱,이종욱,민우성,박종원 대한감염학회 2010 감염과 화학요법 Vol.42 No.3

Background: Micafungin, a potent inhibitor of 1,3-β-D-glucan synthase, is a novel antifungal agent of the echinocandin class. In vitro study showed that micafungin was effective against Aspergillus species as well as Candida species, but clinical data on the prophylactic efficacy against invasive fungal infections (IFIs) other than candidiasis are still lacking. Materials and Methods: We identified 60 consecutive adult hematopoietic stem cell transplantation (HSCT) recipients who received at least 3 doses of micafungin during neutropenic period. Micafungin was started as an alternative in patients who were intolerant or had adverse events (AEs) to primary prophylactic antifungal agents. We retrospectively reviewed the medical records and analyzed the efficacy and safety of micafungin for prophylaxis against IFIs. Results: The patients either had autologous (n=9) or allogeneic (n=51: 1 syngeneic, 24 sibling, 26 unrelated donor) HSCT. Itraconazole oral solution (n=58) was the most frequently used first line antifungal agent for prophylaxis and was administered for median 11 days. The most frequent cause of switch to micafungin was vomiting (n=42). The duration of neutropenia and micafungin administration was median 13 and 12 days, respectively. A successful outcome was achieved in 45 (75%) patients. Empirical antifungal therapy was initiated in 13 (22%) patients. There were 2 cases (3.3%) of breakthrough fungal infections which comprised a probable invasive pulmonary aspergillosis and a possible invasive fungal sinusitis. There was no case of invasive candidiasis. A total of 53 (88%) patients experienced at least one AE regardless of causality during micafungin administration. The most frequent AEs were hypokalemia, vomiting, diarrhea, and elevated serum aspartate aminotransferase or alanine aminotransferase. Among the aforementioned AEs, only 1 case of diarrhea could be classified as a probable relation with micafungin when causality was assessed. There was no AEs that caused discontinuation of micafungin. Conclusions: Micafungin seems to be a safe and effective agent for prophylaxis of IFIs including aspergillosis as well as candidiasis in HSCT recipients. However, further large, prospective, and randomized comparative studies are warranted for aspergillosis.

Lidocaine, Thrombin, Epinephrine 의 항균효과

김진우,이동건,전혜선,김승준,김석찬,안중현,김치홍,권순석,김영균,김관형,문화식,신완식,송정섭,박성학 대한감염학회 2005 감염과 화학요법 Vol.37 No.6

목적 : 기관지내시경 검사에서 흔히 사용되는 국소마취제인 lidocaine과 내시경시 지혈목적으로 사용되는 thrombin과 epinephrine이 각종 균주에 미치는 항균효과에 대해 알아보고자 하였다. 재료 및 방법 : 균주는 가톨릭대학교 성모병원에서 2004년 3월부터 2004년 9월까지 임상검체 에서 동정된 S. aureus, S. pneumoniae, K. pneumoniae, P. aeruginosa 각각 42, 42, 42, 43주를 대상으로 하였다. Lidocaine, thrombin, epinephrine 감수성 검사는 National Committee for Clinical Laboratory Standards (NCCLS, 2002)의 기준에 따랐다. 결과 : Lidocaine은 S. aureus, S. pneumoniae, P. aeruginosa에서 MIC_(50), MIC_(90) 모두 20,000 ㎍/mL 이었다. K. pneumoniae는 각각 10,000 ㎍/mL이었다. Thrombin은 S. aureus와 P. aeruginosa에서 MIC50 500 lU/mL 과 MIC_(90) 500 IU/mL 이상이었고, K. pneumoniae에서는 MIC_(50)과 MIC_(90)이 모두 500 lU/mL이상이었으나 S. pneumoniae에서는 MIC_(50)과 MIC_(90)은 125 IU/mL이었다. Epinephrine은 K. pneumoniae, S. pneumoniae에 대한 MIC_(50), MIC_(90)가 모두 >500 ㎍/mL이었고, S. aureus와 P. aeruginosa에 대한 MIC_(50), MIC_(90)가 모두 500 ㎍/mL이었다. 결론 : 기관지 내시경 검사에서 흔히 쓰이는 lidocaine, thrombin, epinephrine 등의 약제들이 호흡기 질환의 흔한 병원균인 S. aureus, S. pneumoniae, K. pneumoniae, P. aeruginosa 균주들에 대해 항균 효과가 있을 수 있어 세균배양검사에 영향을 미칠 수 있겠다. Background : We performed this stody to find out about antimicrobial effect of lidocaine which is commonly used local anesthetic, and thrombin and epinephrine used for hemostasis during bronchoscopic procedures. Materials and Methods : The microorganisms that were cultured from specimens obtained during bronchoscopy were Staphylococcus aureus (n=42), Streptococcus pneumoniae (n=42), Klebsiella pneumoniae (n=42), and Pseudomonas aeruginosa (n=43) collected from St. Mary's Hospital, from March to Sep 2004 were used for susceptibity testing. Susceptibility to lidocaine, thrombin, and epinephrine were tested according to the National Committee for Clinical Laboratory Standards. Result : MIC_(50) and MIC_(90) of lidocaine for S. aureus, S. pneumoniae, P. aeruginosa were all 20,000 ㎍/mL and that for K. pneumoniae were 10,000 ㎍/mL. MIC_(50) and MIC_(90) of thrombin for both S. aureus and P. aeruginosa was 500 IU/mL and above 500 IU/mL, respectively; that for K. pneumoniae were all above 500 IU/mL and for S. pneumoniae they were 125 IU/mL, MIC_(50) and MIC_(90) of epinephrine for K. pneumoniae and S. pneumoniae were above 500 ㎍/mL; that for S. aureus and P. aeruginosa were 500 ㎍/mL. Conclusion : We observed possible antimicrobial effect of lidocaine, thrombin, and epinephrine in vitro against pathogens such as S. aureus, S. pneumoniae, K. pneumoniae, P. aeruginosa, which are common respiratory microorganisms. The use of these agants could affect the result of bacterial culture.

Smad에 의한 alkaline phosphatase 유전자의 발현 조절기전

김난진,류현모,김현정,김영진,남순현 대한소아치과학회 2001 大韓小兒齒科學會誌 Vol.28 No.2

본 실험은 탁월한 골유도능으로 관심의 대상이 되고 있는 RMP의 세포내 신호 전달자로 알려진 Smad 1과 Smad 5가 조 골세포 초기 분화표지인자인 ALP 유전자의 발현에 미치는 영향 및 그 조절기전을 알아보고자 하였다. BMP 처리 없이도 Smad에 의해 ALP가 발현되는가를 알아보기 위해 Smad 1과 Smad 5가 각각 stably transfection된 C2C12 세포를 3일간 배양후 histochemical assay를 하였고, Smad 1과 Smad 5의 expression vector와 ALP promoter vector를 transient co-transfectiongksgn ALP promoter activity를 측정하였다. Smad에 의한 BMP의 효과를 알아보기 위해서 100ng/ml의 BMP-2를 처리한 군과 처리하지 않은 군으로 나누어 세포를 배양한후 ALP 유전자의 발현을 northern blot analysis로 확인하였다. Smad가 ALP 유전자의 발현을 직접적으로 조절하는가를 알아보기 위여서는 단백질 합성억제제인 cycloheximide를 전처리하여 ALP 유전자의 발현을 northern blot analysis하였다. 이상의 실험결과 다음과 같은 결론을 얻었다. ·Smad 1과 Smad 5가 과발현된 세포에서는 BMP 처리없이도 ALP가 발현된다. ·Smad 1과 Smad 5가 과발현된 세포에서 RMP 처리후 ALP 발현 증가율이 대조군 보다 현저히 높게 나타나 Smad가 BMP 효과를 증가시킨다는 것을 알수있다. ·Smad는 새로운 단백질의 합성을 통해 ALP 유전자를 발현시킨다. Bone morphogenetic proteins(BMPs), members of the transforming growth factor β(TGF-β) superfamily were first identified as the factors that induce ectopic bone formation in vivo, when implanted into muscular tissue. Especially BMP-2 inhibits terminal differentiation of C2C12 myoblasts and converts them into osteoblast lineage cells. In the molecular mechanism of the signal transduction of TGF-β and related factors, intracellular signaling proteins were identified as Smad. In previous study, it has been reported that Smad 1 and Smad 5, which belong to the R-Smad family mediate BMP signaling, were involved in the induction of osteoblast differentiation in C2C12 cells. To understnad the role of Smads involved in osteogenic transdifferentiation in C2C12 cell, in present study, after we stably transfected C2C12 cells with each. Smad(Smad 1, Smad 5) expression vector, cultured for 3 days and stained for alkaline phophatase activity. ALP activity positive cells appeared in the Smad 1, Smad 5 stably transfected cell even in the abscence of BMP. After transiently co-transfected C2C12 cells with each Smad expression vector and ALP promoter, it was excence of MBP. These result suggested that both Smad 1 and Smad 5 were involved in the intracellular BMP signals which induce osteoblast differentiation in C2C12 cells. The effect of BMP on C2C12 cells with Smad 1, Smad 5 transfected were studied by using northern blot analysis. the treatment of BMP upregulated ALP mRNA level in three groups, especially upregulation of ALP was larger in Smad 1, Smad 5 transfected cell than control group. Pretreatment with cycloheximide(10㎍/ml), a protein synthesis inhibitor resulted in blocking the ALP gene expression even in BMP(100ng/ml) treated cell. These results suggested that Smad increased the level of ALP mRNA via the synthesis of a certain transcriptional regulatory protein.

OVERSHOOT를 제거한 개선된 큐빅 스플라인 보간법에 관한 연구

김성현,김진만,조상현,주동현,김두영 東亞大學校附設情報技術硏究所 2005 情報技術硏究所論文誌 Vol.12 No.2

This study presents the improvement of cubic spline interpolation without overshoot. The o-riginal cubic spline interpolation has overshoots between two data. They are noises in still image zoom because of getting out of the pixel value. But the improvement of cubic spline in-terpolation removes them and proffers better quality still image than other interpolations.

Mouse의 치아 발육시 Runx2의 발현 양상

김태완,류현모,남순현,김영진,김현정 大韓小兒齒科學會 2004 大韓小兒齒科學會誌 Vol.31 No.4

Runx2는 runt gene family에 속하는 전사조절 인자로써 뼈의 형성과 골아세포의 분화에 중요한 역할을 담당하고 있다. Runx2-haploinsufficency는 쇄골의 저형성 및 두개 봉합의 지연을 특징으로 하는 쇄골두개 이형성증을 일으키며, 치아에 있어서는 법랑질의 저형성, 영구치 맹출지연 등을 보인다. 이에, 치아의 발육 및 맹출에 미치는 Runx2의 영향을 알아보기 위해 in situ hybridization 방법으로 태생 1, 4, 7, 14. 21일 된 쥐의 하악 및 제1대구치를 사용하여 실험을 실시하였다. Runx2-full length는 태생 1일과 4일에 치낭 및 주위조직에 보이지만 Runx2-typeⅡ는 보이지 않았다. Runx2-full length는 태생 7일에 치관 교합면 부위의 법랑모세포에 발현하였고, 1주일 후인 태생 14일에는 백악법랑경계 상방의 치관인 접면 법랑모세포에서 발현되었다. 이에 반해 Runx2-typeⅡ는 법랑모세포에서 발현하지 않았다. 또한 태생 21일에서는 두가지 이성질체가 모두 하악골에서 발현을 보였다. 이런 결과를 종합해볼 때, Runx2-full length는 치아의 맹출과 연관이 있으며, 법랑모세포의 분화 및 이로 인한 법랑질형성에 영향을 주지만 Runx2-typeⅡ는 하악골의 형성에만 영향을 미치는 것으로 사료된다. Runx2 is a transcription factor in homologous with Drosophila runt gene and it is essential for bone formation during embryogenesis and a critical gene for osteoblast differentiation and osteoblast function. Runx2-haploinsufficency causes cleidocranial dysplasia (CCD). CCD is an autosomal-dominant inherited disorder characterized by hypoplastic clevicle and delayed ossification in fontanelles and wormian bones. Dental defects are possibly shown to CCD patients : multiple supernumerary teeth, irregular and compressed permanent tooth crowns, hypoplastic and hypomineralized defects in enamel and dentin, an excess of epithelial root remnants, the absence of cellular cementum, and abnormally shaped roots. In addition, delayed eruption of the secondary dentition is a constant finding. The aim of this study is to evaluate the role of Runx2 in the tooth development and eruption through analyzing the expression pattern of Runx2 by in situ hybridization during crown (late bell stage) and root formation of tooth, using postnatal day 1, 4, 7, 14 and 21 mice mandibular molar teeth. mRNA of Runx2-full length is expressed in dental follicle and surrounding tissue at postnatal day1 and 4. At post-natal day 7, it is expressed in ameloblasts of occlusal surface of enamel and bone area surrounding the tooth. In comparison with previous stage, at postnatal day 14, it is expressed in ameloblasts of proximal surface of enamel. At postnatal day 21 it's expression is observed only in bone area. mRNA of Runx2-typeⅡ is not expressed At postnatal day 1 and 7. At postnatal day 14 and 21, it's expression is observed in the bone area. In this study, we suggest that Runx2 have a relation of ameloblasts differentiation and an important role to tooth eruption made by dental follicle during intraosseous eruption stage. Also we can confirm that Runx2 has a role to bone formation.