본 연구는 소의 초기 난할 단계인 2 또는 4세포기 수정란의 특정 분할구가 배반포 단계의 내부 세포괴(Inner Cell Mass) 와 영양막 세포(Trophectoderm cells)로의 발달 운명이 미리 정해지는 지를 확인하기 위해 실시되었다. 먼저 생쥐의 체내 수정란과 소의 체외 수정란에서 배반포의 영양막 세포에서만 특이적으로 발현하는 cdx2 단백질의 발현 양상을 조사하였다. 또한, 소의 경우 2세포기와 4세포기가 내부 세포괴와 영양막 세포로 나눠지는 시점인지를 조사하기 위해 2 또는 4세포기의 특정 분할구에Dextran의 주입 실험과 분할구 제거 실험을 통해 ICM과 TE 형성을 확인하였다. cdx2의 발현 경향은 생쥐와 소의 2세포기일 때 대칭과 비대칭적으로 발현되는 것을 확인하였다. 생쥐의 4, 8세포기 및 상실배기에서는 분할구 전체에서 발현되었으나, 소 수정란의 분할구에서는 전체 또는 부분적으로 발현되었다. 또한, 생쥐와 소의 배반포기에서는 영양막 세포에서만 발현이 되는 것을 확인하였다. 소 수정란의 2세포기와 4세포기 단계에서 특정 분할구에 주입된 Dextran은 배반포의 내부 세포괴와 영양막 세포의 양쪽에 분포된 것을 관찰할 수 있었다. 2세포기 단계에서 하나의 분할구가 제거된 수정란 역시 ICM 및 TE 세포를 지닌 정상 배반포로 발달함을 확인하였다. 따라서 본 연구 결과는 영양막 세포에서만 특이적으로 발현하는 cdx2의 발현이 2 또는 4세포기 단계 소 수정란에서는 특별한 차이를 보이지 않으며, 궁극적으로 난할 초기에는 ICM과 TE 세포로의 운명이 결정되지 않는다는 것을 보여준다.

본 연구는 사람 H-transferase가 과발현하는 돼지 체세포주를 개발하는데 있다. 돼지 세포에 사람 H-transferase 유전자를 발현시키는 것은 이종간 장기 이식에 있어서 초급성 거부 반응을 방지하기 위한 한 가지 방법이다. 본 연구에서는 과발현 벡터를 구축하기 위하여 사람 H-transferase을 HepG2 세포로부터 동정하였으며, 이 유전자를 CMV promoter를 이용하여 발현할 수 있도록 포유동물 발현 벡터인 pRc/CMV 벡터에 삽입하였다. 또한, 돼지 산자의 귀 세포를 이용하여 체세포를 수립한 후 jetPEI DNA transfection reagent를 이용하여 벡터를 도입하였고, 300 μg/ml의 G418로 12일간 선별하였다. PCR을 이용하여 선별된 colony들을 분석한 결과, 벡터가 도입되었음을 확인하였고, RT-PCR을 이용하여 사람 H-transferase mRNA가 발현하는 것을 확인하였다. 본 연구에서 확립된 세포주는 사람 H-transferase가 과발현하는 형질 전환 돼지의 생산에 이용될 수 있을 것으로 생각된다.

체세포의 배양 방법은 체세포 핵이식에 의한 형질 전환 돼지 생산에 있어서 중요한 요인 중 하나이다. 본 연구에서는 돼지 태아 섬유 아세포의 효율적인 배양 방법을 수려하였다. 돼지 태아 섬유 아세포는 임신 33일째 태아로부터 제조하였으며, 돼지 태아 섬유아세포의 증식을 혈청과 배지 종류별로 분석하였다. 그 결과, 15% ES screened FBS가 포함된 DMEM 배지에서의 배양은 15% FBS보다 세포수의 증가가 훨씬 더 빠르게 나타났다. 또한, 태아

임신의 성립 및 유지에 중요한 역할을 하는 자궁내막세포에 통로의 존재를 확인하기 위하여 본 연구를 수행하였다. 통로는 일반적으로 중추신경계에 풍부하게 존재하면서 세포의 안정막 전압을 유지시킨다. 역전사 중합 효소 중합 반응과 면역 세포 화학 염색 방법을 이용하여 자궁내막세포에 존재하는 통로를 조사한 결과, TASK-1, TASK-3, TREK-1, TREK-2 및 TRAAK의 발현이 확인되었다. TASK-3와 TREK-1은 핵을 포함한 세포 전역에 발현

본 연구는 체세포를 이용하여 생산된 복제 한우 수소의 번식능력을 검토하기 위해 실시하였다. 복제 한우 수소(C-38 및 C-39) 또는 일반 한우 종모우로부터 정액을 채취하여 정자의 수 및 동결 전후의 생존성 등을 살펴보았으며, 정자의 운동성 등은 computer assisted sperm analysis(CASA)를 이용하여 측정하였다. 또한, 이들의 수정 능력을 확인하기 위하여 체외수정과 인공수정을 각각 실시하였다. 정액 성상에서는 복제 수소들과 일반 종모우 간에 정액의 양, 정자의 농도 및 동결융해 후의 생존성 등에서 차이가 나타나지 않았다. CASA를 이용한 분석에서 운동성, 곡선 운동 속도(VCL), 직선 운동 속도(VSL) 및 평균 진행 속도(VAP) 등은 복제 수소의 정액이 일반 종모우의 정액에 비하여 유의적으로 높았다(p<0.05). 체외수정에 따른 수정란의 분화율 및 배반포로의 발달율은 복제 수소와 일반 종모우 간에 차이가 나타나지 않았다. 복제소 정액(C-38)을 이용하여 인공수정을 한 5두의 체세포 복제 대리모에서 암수 각각 한 두씩의 건강한 복제 후대 송아지 2두를 생산하였다. 이상의 결과를 종합하여 보면, 실험에 공시된 복제 수소 개체 간의 차이가 나타나기는 하였지만, 복제 수소는 정액 성상과 정자의 운동성 등에서 일반 종모우와 차이가 없었으며. 또한 인공수정을 통해 송아지를 생산함으로써 정상적인 번식능력이 있음을 확인하였다.

Osteoblastoma is a benign bone forming neoplasm most commonly occurring in the vertebrae, long bones, but the jaw involvement is very rare events. When localized close to the periapical region or laterally to the roots of the t eeth, the lesion is easily confused with inflammatory periapical pathosis. In this article, we report our experience with a case of osteoblastoma located in the periapical region that was initially misdiagnosed as an inflammatory periapical pathosis, which led to unnecessary dental treatment and a delayed diagnosis. This case demonstrates the importance of proper diagnosis and treatment planning when one is dealing with radiolucent lesions in the periapical area Therefore, this case stresses the importance of biopsy of periapical lesions that do not respond to endodontic treatment or are otherwise S USpl CIOUS.

The purpose of this study was to examine the effects of vi tamin D3 and 1'etinoic acid(RA) on the human mesenchymal stem ce!ls(MSC) g1'owth and osteogenic differentiations. Cell proliferation, mineralization, cell cycle, expression of cell cycle regu l atOJγ proteins and markers fo1' osteogenic differenatiaiton were determined by MTI assay, mineralization assay, flow cytomet1'Y‘ and Western blot analysis, respectively. Cell viability was dec1'ease by each vitamin D3 and RA added to MSC. it was more decrease by vitamin D3 and RA. Mineralized nodule formation revealed similar expression pattern with positive cont rol group at vitamin D3 and RA mixed add to MSC. At vitamin D3 and RA mixed add to MSC after 7 days of incubation was increase G1 s tage. after 21 days of incubation was inhibit cell cycle prog1'ess by inc1'ease of sub-G1 Treatment vitamin D3 to MSC inhibits p53 and p21, but inc1'ease pRb. RA inhibit p53, but increase p21 and pRb, vitamin D3 plus RA group was same as added RA group. so two vitamin was effect to inhibited cell growth each different mechanism. Expression of BMP-2 protein was prominent in osteogonic supplement treated g1'oup of MSC at 2 weeks cultivation days, but vi tamin D3 treatment decreased BMP-2 expression rather than in (+) control group. BSP protein was notably increased in the OS compa red to positive controls at 2 weeks cultivation, but similar to that of vitamin D3 group t1'eatment group and was least expressed in plus RA mixed group, at 3 weeks, BSP expression was similar to 1'esult of 2 weeks Collectively, these results shows that vitamin D3 and RA have diffe1'ential effects on the MSCs g1'owth and differ entia tion 211

Disruption of cell - matrix attachment results in a loss of prosurvival signals and culminates in programmed cell death, referred to as anoikis , Apoptosis signal- regulating kinase 1(ASKl)/MKK5 is a ubiquitously expressed enzyme that acti vates c-Jun N-terminal kinase/stress-activated protein kinase JNK/SAPK and p38 pathways by direct site specific Ser/Thr phosphoryl ation of their respective MKKs-MKK4/MKK7 for JNK and MKK3/MKK6 for p38 kinases, The kinase activity of ASKl is stimulated by a variety of death signals, including TNF, Fas ligation, reactive oxygen species, and antineoplastic agents , The aim of this study was to investigate the relative importance of ASKl in anOlkls 1n the present study cells which lost their adhesion showed higher rate of cell death in compared to cells which maintained anchorage. 1nterestingly the res ult showed that suspended cells expressing ASK1 were more susceptible to anoikis than suspended cells having no ASK1 1n addition, cellu lar attachment seems to have significant effect on ASKl activity and p38 MAPK protein rather than serum stimulation

Cytokines play a vital role in the host immune response by regulating the development and function of im munocompetent ce11s One immunomodulatory agent that has received attention in oncology research recently is interleukin - lO(IL-lO). IL-IO inhibi ted tumor antigen presenta tion and induced energy in T lymphocytes that had been s timu lated by autologous MHC class II positive tumor ce11s Patients with head and neck cancer have been shown to exhibit profound irnmunosuppression. The mechani sm by which tumor ce11s alter immunological function in the host is poorly understood. Recently. production of biological active IL- IO was confirmed in ovar‘ian cancer, melanoma, skin cancel‘ & head and neck cancer, suggesting that IL- lO reduces the function of tumor infiltrating lymphocytes and contributes to the tumor growth. IL-IO expression has not been examined extensively in human oral cancer and has not yet been cla rified. The purpose of t his study were to investigate IL-IO mRNA and protein expression in NHOK, IHOK and oral squamous ce11 carcinoma(OSCC) ce11 line by RT-PCR and irnmunoslot blotting, and to apply its results to examine its thera peutic significance for oraJ cancers. Cultured NHOK showed a lower level of IL-IO mRNA and protein expression than cultured IHOK and HN 22 OSCC cell line under pre and postconfluency. HN 22 OSCC cell line under pre and postconfl u ency. showed the highest level of IL-I0 Cul tured IHOK showing a intermediate expression of IL- IO could be as a vaJ u a bJe marker for oral carci nogenesis ste p. During the terminal differentiation of a11 the ce11 lines, IL- IO ex pression was significantly unchangeabl e. IL- IO mRNA expression of a11 the ce11 lines was consistent with IL-10 protein expression. It suggested that IL- lO expression might play an important role in oral carcinogenesis and IHOK could be a valuable marker for oral carcinogenesis step. And aJso IL- 10 related gene may be future targets for gene discovery and possi bJy therapeutic intervention

Desmoplastic ameloblastoma(DA) is histologically characterized by extensive stromal collagenization or desmoplasia. ln this study, anti-cytokeratin 8/18, 13, 19 for pathogenesis as well as anti-PCNA for cellular proliferation, were used to det ect the expression of these proteins in the desmoplastic ameloblastoma Basal layers of tumor nest were negatively stained by CKl3, while suprabasal and inner cells were positive for CK13. CK8/18 and CK 19 was negatively stained in the peripheral portion of tumor nest in DA, whereas CK 8/18 was in central portion and CKl9 was positive in the su prabasal and some of central portion of the cel l nest. PCNA index of DA was 60 ::!: 14.6% to 95 ::!: 17 .2%. The peripheral tumor cells of the islets presented higher PCNA labeling index, while some cells in the central area of foll icle containing squamous like cells also presented negative PCNA labeling index. Especially tumor islands showed higher PCNA index than in main tumor mass. lt suggested that desmoplastic ameloblastoma might be composed of many different tumor cell types‘ and have hi gher pr이 ife r a ting activity in tumor islands of the desmoplastic stroma

The support mechanisms that are involved in lymph node metastasis of oral squamous cell carcinoma remain largely unknown. Recent studies have demonstrated that tumor cells express chemokine receptors and use chemokines to metastasize to the target organ in many malignancies in humans There are few reports about the correlation between chemokin receptor CXCR-4 expression and clinicopathologic factors in oral squamous cell carcinomas. The object of this study was to evaluate the availabili ty of CXCR-4 expression as prognostic marker through correlation analysis of CXCR-4 expression in oral sq uamous cell carcinoma and its r elation to clinocopathologic factors and PCNA index. 80 we investigated CXCR-4 expression of 74 oral squamous cell carcinomas by immunohistochemistry. 44 out of 74 cases(59. 5%) showed CXCH-4 positive and 30 sampl es(40.5%) showed CXCH-4 negative. CXCH-4 expression showed statistically sig nificant correlation wi th lymph node metastasis(p=0.026) ‘ PCNA index (p=0.003) , survial rate(p=0.0003). From the results , it was suggested CXCR-4 oxpression might be useful a prognostic marker in oral squamous cell carClllomas

To determine the optimal concentration of fetal bovine serum (FBS) on the growth of insect cells and the multiplicity of viruses, the growth of cells (Sf21 and Bm5) and viruses were examined on the various concentrations of FBS. In view of the viability, growth speed, proliferation of cells and the amount of FBS, the most proper concentration for the cell culture were 7% and 5% for Sf21 and Bm5, respectively. The multiplicity of viruses at the various concentrations of FBS was similar in both cell lines at 5 days post-infection (p.i.). However, it differed significantly at 2 and 3 days p.i. The proper concentration of FBS were 10% and 3% for Sf21 at 2 and 3 days p.i., respectively, and 5% for Bm5 at both 2 and 3 days p. i. These results suggested that the optimal concentration of FBS should be determined according to the used cell lines and viruses for their optimum production.

We have examined the effect of NO donor, S-nitl‘ oso-N-acetyl-DL-penicillamine(SNAP) on heme oxygenase-1 (HQ-l) ex pression in human oral immortalized & malignant keratinocytes, and investigated in the control of keratinocyte proliferation evidence tha t HO-1 cou ld be involved in a low dose of NO, NO inhibitor, HOinducer, and HO inhibitor medi ated cytoprotect ion against cytotoxi city induced by a high dose of NO Oral keratinocyte growth inhibitory or anti-proliferative effects were exerted by with SNAP and hemin in a dose- and cul tivation time dependent manner The level of HQ-1 protein was increased in all cell types after exposure hernin dose, and the hemin induced HQ-1 protein achieved at higher maximum level by 12 hrs in all kind of cells , The pretreatment of cells with 0, 2 μ M SNAP reduced 1 mM SNAP-induced death in IHOK and HN4 cells , These cytoprotective effects on high dose of NO induced HQ-1 expresion and cell ular toxicity were blocked by low dose of SNAP, HCB, and ZnPP IX supporting the involvement of HQ-1 in high dose NO induced growth arrest or cell death, But these cytoprotection pattern is different from immortalized and malignant keratinocytes , These results indirectly demonstrate that HQ-1 could be involved in cytoprotection by NO priming against high dose NO induced cytotoxicity in immortalized and maigla nt oral keratinocytes, Thus, HQ-1 might be an important cellular target of NO donor, with clinical implications for the pre vention of inJlammatory di seases and anti-tumor immunity

본 연구에서 돼지 난포란에서 채취된 난모 세포들을 체외성숙 후 형태적으로 선별하거나 극체 방출란을 선별하여 활성화 처리 후 48시간째에 분할란을 선별할 때 배발달율이 어느정도 향상되는지를 검토하였다. 난모 세포를 48시간 성숙 배양 후 형태적 선별과 극체의 방출 유무를 검사하고, 선별된 난모 세포들을 시간 추가 배양한 후 7% ethanol로 활성화시키고 cytochalasin B에 5시간 노출 후 PZM-5 배 양액으로 7일간 배양하였으며, 배양 중

본 연구는 재래 산양의 체세포 핵이식에 의하여 생산한 복제 산양(진순이)의 조직으로부터 공여 핵을 배양하여 다시 핵이식을 실시하여 재복제에 따른 융합율과 분할율, 이식 후의 수태율 등을 조사하여 재복제 가능성 여부를 검토하기 위하여 실시하였다. 공여 세포는 귀 유래 섬유아세포를 분리 배양하여 사용하였으며, 체내 성숙 난자는 성숙한 미경산 재래 산양에 과배란을 유기하여 외과적인 방법으로 난관 관류를 통해 회수하여 핵이식을 실시하였다. 핵이식란의 융합은 전