• 대한한의학회지
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• 제20권3호
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• pp.54-65
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• 1999

Objectives: Hepatoma is a very serious disease in Korea and vvorldwiclc. Hepatitis B vims (HBV) has proved the most significant cause of hepatoma. We canied out this study to investigate the effect of Acanthopanax sessilifloms on inhibiting cell proliferation and DNA synthesis in HepG2.2.15 cell line and on inhibiting phosphorilation of oncogene (MAP kinase) in NIT/3T3-HBx ceIl. Methods: To investigate the anti-cancer effect of Acanthopanax sessiliflorus, we did the CellTiter 96 Aqueous Non-radioactive Cell Proliferation assay (Promega); MTS/PMS assay, [$^3H$]-thymicline incorporation assay, and we measured the gene expression through westem blotting. Results: Acanthopanax sessiliflorus showed an inhibiting effect on the increase of HepG2.2.15 in the NTS/PMS assay. It also showed an inhibiting effect on DNA synthesis of HepG2.2.15 in the [$^3H$]-thymidine incorporation assay. Acanthopanax sessiliflorus showed an inhibiting effect of phosphorilation of MAP kinase in HBV - X genes. too. Conclusions: The results suggested that this herb had an anti cancer effect. We may discover an effective anti-cancer herb medicine through further studies on this herb medicine.

• 한국환경보건학회지
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• 제24권2호
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• pp.126-133
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• 1998

Effects of B(a)P on preimplantation mouse embryos in vitro were studied. Preimplantation mouse embryos were exposed to a concentration of 0.3, 1, 3 and 10 $\mu$M B(a)P for 72 hrs. The toxicological effects of B(a)P were evaluated by morphological observation of embryos up to the blastocyst stage, and by measuring DNA, RNA and protein synthesis by radioactive precursor incorporation. At 1 $\mu$M B(a)P did not affect preimplantation development but interfered with hatching and ICM formation. Suppressing effect of ICM formation was dose dependent. At the eight cell stage, the developmental rate was decreased at above 3 $\mu$M of B(a)P. At the blastocyst stage, attachment and trophoblast outgrowth were diminished at the 10 $\mu$M of B(a)P and ICM formation was decreased at 1 $\mu$M of B(a)P. Inner cell number of blastocyst was decreased dose dependently. So, number of ICM was one of the most sensitive and toxicological end point. The RNA incorporation rate of 0.1 $\mu ^3$H-uridine was dosedependent and the protein incroporation of 0.5 $\mu Ci ^{35}$S-methionine showed a significant decrease after 48 hrs. But the DNA incorporation rate of methyl-$^3$H thymidine was not affected. Our results suggested that B(a)P did not affect the DNA replication but transcription was inhibited by dose dependent manner. There delay of development during the blastocyst stage was mainly due to the inhibition of RNA synthesis followed by protein synthesis.

• 대한수의학회지
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• 제26권2호
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• pp.245-249
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• 1986

The present study has been carried out to investigate the optimal condition on the blastogenesis of guinea pig lymphocytes. A microculture system in conjunction with a semiautomatic multiple sample harvester(SAMSH) was used to study the in vito optimal condition of guinea pig lymphocytes. Data were presented to show many variables that are Involved in studying the responses of guinea pig lymphocyte in a microculture system to the stimulation of Concanavalin A(Con A) and lipopolysaccharide (LPS). Analysis indicated that the conditions for optimal Con A as measured by incorporation of $^3H$-TdR include : (1) use of RPMI-1640 as culture medium, (2) use of $6{\mu}g$ of Con A, per culture, (3) use of $1{\times}10^6$ cells per culture. Conditions for optimal stimulation with LPS mitogen were similar to those used for Con A.

• Toxicological Research
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• 제19권4호
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• pp.293-296
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• 2003

The effect of the hexane extract of Curcuma zedoaria roots and its solvent fractions were investigated on the proliferation of SiHa, SNU-1 and HepG2 cell lines. Among those fractions, final fraction H2-3-1 and H2-3-3 showed cytotoxic effect on SiHa and HepG2 cell lines. The hallmark of apoptosis, DNA fragmentation, also appeared in the final fractions H2-3-1 and H2-3-3 after 24h treatment in SiHa cell line. Furthermore, those fractions were shown to be able to induce cell death in $[^3H]$thymidine incorporation test. These two fractions, H2-3-1 and H2-3-3 were determined as (-)-$\alpha$-curcumene and $\beta$-tumerone respectively by NMR and mass spectrum. From these results, it is speculated that te hexane extract of Curcuma zedoaria is necessary for further studies as a potent inhibitor of the growth of cancer cells.

• 대한치과교정학회지
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• 제22권2호
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• pp.309-325
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• 1992

The purpose of this study was to evaluate the effect of intrinsic factor and extrinsic factor for growth of the mandibular condylar cartilage of 4 day-old rats in a serum-free medium for 1, 4, 7, 14 days. They were compared with normal growth in vivo and with growth of spheno-occipital synchondrosis in serum-free medium. The cellular kinetics of cartilages were evaluated by auto-radiography of tritiated thymidine. 1. Condylar cartilage was enlarged with rounded head on day 14 of experiment while in vivo the rounded-headed shape changed into functionally flattened appearance. 2. On day 14 of experiment, a severe reduction of the proliferative zone and a considerable increase of the hypertrophic zone were observed while in normal control group endochondrol bone formation and bone marrow were observed. 3. The proliferative activity in the proliferative zone of condylar cartilage detected by $^3H-thymidine$ incorporation was lower than that of normal control group and decreased more than that of spheno-occipital synchondrosis, but it continued during the 14 days of culture. 4. The continued maintenance of condylar cartilage and morphologic change were disturbed in this culture system, but cell division within the proliferative zone was continued and probably linked to intrinsic factor.

• IMMUNE NETWORK
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• 제2권1호
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• pp.19-24
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• 2002

Background: Salvia miltiorrhiza (SM), a traditional oriental medicine, has been reported to have anti-tumor properties, but its exact mechanism remains to be elucidated. In this study, we investigated several of the molecular events that occur in human breast carcinoma MCF-7 cells and human pulmonary adenocarcinoma A549 cells. Methods: For this purpose, we evaluated the growth-inhibitory effect of SM in association with the expressions of p53, p21, cyclin D1, and pRb, which are known to be involved in cell cycle arrest. The extent of thymidine incorporation was also examined to assess G1/S phase cell cycle arrest in both cells by $^3H$-thymidine incorporation. Results: Our results show that SM inhibits the growth and the proliferation of MCF-7 and A549 cells. Furthermore, we also observed increased expression of p21 via a p53-dependent pathway in both cell lines after treating with SM. In addition, treatment with SM for 24 hours caused the suppression of hyperphosphorylated retinoblastoma protein (pRb) expression and the dephosphorylation of pRb. Conclusion: These findings suggest that the growth inhibitory and the anti-proliferation effects of SM on MCF-7 cells and A549 cells are mediated via the decreased expression and dephosphorylation of pRB by p21 up-regulation in a p53-dependent manner. To the best of our knowledge, this study is the first to report upon the molecular mechanisms involved in SM-induced tumor cell growth inhibition.

• 한국해양바이오학회지
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• 제1권2호
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• pp.119-125
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• 2006

어류의 insulin-like growth factor-I (IGF-I)의 생화학적 작용기작을 연구하기 위하여 한국산 조피볼락의 IGF-I cDNA 유전자 cloning을 행하였다. 완전한 cDNA 유전자 염기서열은 PCR과 RACE 방법을 통하여 얻어진 DNA로부터 결과를 얻을수 있었다. 결정된 IGF-I의 염기서열은 flounder, chinook salmon, human IGF-I의 염기서열과 비교한 결과 각각 93.6%, 90.7%, 85.4%의 높은 상동성을 보였다. 생화학적으로 활성이 있는 재조합 IGF-I을 얻기 위하여 IGF-I의 B-C-A-D domain 부분을 PCR로 얻은 뒤 E. coli BL21(DE3)에 넣어 overexpression 시켰다. Ni-NTA colummn을 사용하여 순수한 재조합 단백질을 정제할수 있었다. 정제된 단백질은 SDS-PAGE 상에서 7 kDa의 단일 band를 보여 주었으며 [$^3H$]-thymidine 결합정도를 측정하는 방법으로 활성을 가지고 있음을 확인할수 있었다.

• Journal of Periodontal and Implant Science
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• 제24권3호
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• pp.572-580
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• 1994

The purpose of the present study was to evaluate the effect of platelet - derived growth factor(PDGF) - BB and insulin - like growth factor(IGF) - 1, Centella Asiatica, and Zea Mays L. on the mitogenic activity of PDL cells from healthy and RPP patients. Combination of PDGF - BB and IGF - 1, Centella Asiatica, and Zea Mays L. were treated on PDL cells and the mitogenic effects were meaured by quantitative assay of methyl - $^3H$ - thymidine incorporation during DNA synthesis. Combination of PDGF - BB and IGF - 1 enhenced the mitogenic effects of both healthy and RPP PDL cells, however, the effect was less pronounced on RPP PDL cells. In cases of Centella Asiatica and Zea Mays L., no mitogenic effect on healthy PDL cells could be noticed.

• Journal of Ginseng Research
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• 제16권3호
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• pp.210-216
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• 1992

The effect of saponin extracted from Panax grneng CA Meyer on DNA repair and replicative DNA synthesis were examined in CHO-Kl cells cotreated with benzo(a)pyrene and rat liver S-15 fraction. The DNA strand breaks inititated by benzo(a)pyrene metabolites were measured by alkaline election technique. The addition of ginseng saponin to the culture media resulted in decrease of benzo(a)pyrene-induced DNA strand breaks, and restored the suppressed-semiconservative-DNA-synthesis by the carcinogen. DNA repair synthesis in the damaged cells was also elevated by the ginseng treatment when the repairing activites were measured for the (3H)-thymidine incorporation into the carcinogen damaged cellular DNk Comparative analysis of DNA-adduces of benzo(a)pyrene metabortes in microsomes suggested that ginseng saponin treatment in rats reduced the formation of electrophilic metabolites of benzo (a)-pyrene in the rat liver microsomes.

• The Korean Journal of Physiology
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• 제8권1호
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• pp.27-30
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• 1974

인삼이 마우스의 간조직 방사능지수에 미치는 영향을 알아보기 위하여 몸무게 $18{\sim}20\;g$ 되는 마우스 수컷 30마리를 사용하여 다음과 같은 실험을 실시하였다. 인삼주정추출액은 고려인삼 300g을 95% 에칠 알코올로 중탕남비 위에서 약 300시간 동안 추출하여 얻은 54.2 g의 흑갈색 추출물을 생리적 식염수 1 ml 속에 이 추출물 4 mg 이 포함되도록 용해시켜 만들었다. 인삼군에는 5일 동안 매일 같은 시각에 인삼주정추출액을 몸무게 10 g에 대하여 0.05 ml씩 등의 피하에 주사하였으며 식염수군에는 몸무게 10 g에 대하여 0.05 ml 씩 피하에 주사하였다. 인삼주정추출액 혹은 식염수 투여가 시작된지 제 5일째 되는 날에 해당 약물을 주사한 다음 2시간 후에 $[^3H]$ thymidine을 단 1회에 복강 속에 주사하고 이어서 이들 동물을 1, 10 및 24시간 후에 한번에 5마리씩 도살하여 간조직을 적출하고 자기장사법을 이용하여 방사능지수를 계수한 바 다음과 같은 결과를 얻었다. 1. $[^3H]$ thymidine 주사 후 1, 10 및 24시간만에 관찰된 식염수군의 간조직 방사능지수는 각각 $3.23{\pm}0.23,\;5.20{\pm}0.21$ 및 $6.00{\pm}0.30$이었다. 2. 인삼군의 간조직 방사능지수는 1, 10 및 24시간 값이 각각 $4.02{\pm}0.33,\;6.32{\pm}0.32$ 및 $7.42{\pm}0.35$으로 나타났다. 그러므로 인삼군의 간조직 방사능지수는 식염수군의 해당 값보다 현저하게 증가되었다. 이상의 결과로 미루어 보아 인삼은 마우스의 간조직 DNA 합성능을 촉진시킨다고 믿어진다.세에 다발하는 경향을 보이고, 오히려 연령이 높아질수록 질병이환율이 감소하는 경향을 보이는 반면, 안이비인후과질환만이 점진적 증가를 보여 대조를 이루었다. 4) 가족력과 질환과의 상관관계를 살펴보면, 중풍환자의 25.0%가 중풍의 가족력을, 고혈압환자의 11.6%가 고혈압의 가족력을 가지고 있는 것으로 나타나 가족력과의 상관관계가 매우 높게 나타남을 알 수 있었고, 그밖에 결핵은 5.6%, 당뇨병은 2.6%가 가족력을 가지고 있는 것으로 나타났다. 5) 음주와 질환과의 상관관계를 살펴보면, 호흡기질환의 36.4%, 고혈압의 34.7%, 심장병의 33.3%, 소화기질환의 28.4%가 음주를 하는 것으로 나타났으나 음주의 정도는 조사되지 않아 음주와 질환과의 필연적 관계는 알 수 없었다. 6) 흡연과 질환과의 상관관계를 살펴보면, 호흡기질환의 44.1%, 심장병의 38.0%, 고혈압의 29.8%가 흡연을 하는 것으로 나타났으나 흡연의 정도는 조사되지 않아 흡연과 질환의 필연적 관계는 알 수 없었다. 7) 사상체질의 분포는 태음인(太陰人) 44.6%, 소양인(少陽人) 30.7%, 소음인(少陰人) 24.7%, 태양인(太陽人) 0.0%의 순으로 나타났고, 체질별 질환빈도는 모든 체질에서 운동기계 질환과 소화기계 질환의 이환율을 높았는데, 태음인에서는 순환기계질환(50.0%), 호흡기계질환 (23.1%)이 타체질에 비해 높게 나타났고, 소양인에서는 운동기계질환(77.7%), 난청등 안이비인후과 질환(12.2%)이 타체질에 비하여 높게 나타났고, 소음인에 있어서는 소화기계 질환(65.8%)이 타체질에 비해 높게 나타났다. 8) 양방의 1차검진상 이상소견자의 빈도와 한양방종합판정에 의한 질환의 빈도 사이에는 상당한 차이가 있는 것으로 나타났는데, 이는 노인질환의 특성상 건강검진에 한방의 참여가 필요한 이유가 되는