The present study reports the protective properties of a total methanol extract of B. platyphylla var. japonica against ultraviolet (UV)-C irradiation. Pretreatment of Chinese hamster fibroblast (V79-4) cells with a total methanol extract significantly increased cell survival following 300J/m² of UV-C irradiation. The total methanol extract was further fractionated into 5 fractions: n-hexane, dichloromethane, ethylacetate, n-butanol and water fractions. Among these fractions, B. platyphylla var. japonica ethylacetate, butanol and water fractions showed significant protective effects against the cellular damage induced by UV-C irradiation. In order to elucidate the mechanism underlying this protective effect, DPPH (Editor note: abbreviations should be spelled out at first use.) radical scavenging and lipid peroxidation inhibitory activity were measured. Significant radical scavenging and lipid peroxidation inhibitory activities were observed for the ethylacetate fraction. In summary, the present data demonstrate that an extract of B. platyphylla var. japonica has a significant protective effect against UV-C irradiation. The underlying mechanism of this protective effect may involve radical scavenging and inhibition of lipid peroxidation by the B. platyphylla var. japonica extract.
The Chinese Hamster Ovarian cells CHOK1 are one of the most extensively used cells for the evaluation of gene expression and toxicology. However, these cells are frequently used for biomedical research without consideration of their cytogenetic characteristics. Therefore, we carried out to investigate the karyologic profiles, the frequency and type of chromosome aberration, and the distribution of telomeric DNA on chromosomes of the CHOK1 cells. The GTGbanding and fluorescence in situ hybridization on CHOK1 cells were performed to characterize the karyotype and the distribution of telomeric DNA. The present study revealed that the chromosome modal number of CHOK1 cells was 2n=20; eight chromosomes appeared to be identical with those of the normal Chinese hamster, whereas the remaining 12 chromosomes were shown to be translocated, deleted, inversed, or rearranged from Chinese hamster chromosomes. The telomeric DNA on CHOK1 chromosomes was intensively distributed at the centromeres rather than the ends of chromosomes. In addition, three chromosomes had interstitial telomeres and one marker chromosome entirely consisted of telomeric DNAs. The frequency and type of chromosome aberrations in CHOK1 cells were examined. Of the 822 metaphase spreads, 68 (8.3%) cells resulted in chromosome aberrations of which the chromosome breakage was the most frequently occurred.
The inhibition of cell recovery might be proceeded via either the damage of the mechanism of the recovery itself or via the formation of irreversible damage which could not be repaired at all. Both these processes may take place at the same time. Any of t
Cyclooxygenase- 2 (COX-2) is an inducible enzyme that is not found in normal conditions,. but is induced by a varie ty of pathophysiologic conditions of tissues by growth factors. inflammatory stimuli. oncogenes and tumor promoters, COX-2 is upregulated in a number of epithelial cancers. including in oral premalignant and malignant lesions, The mode of action of COX-2 in carcinogenesis may include mutiple mechanisms that may be acting at different stages of malignant disease, In this study. the expression of COX- 2 protein was assessed quantitatively 없d qualitatively by immunohistochemistry during DMBA-induced hamster buccal pouch carcinogenesis, The immunoreactivity for COX-2 protein increased as the tissue passed from hyperplasia to dysplasia and SCC, The highest mean expression was SCC at 14 week, The differences between COX-2 expression in the normal and that the dysplastic and carcinomatous lesions was statistically significant, In addition. the mean values of COX -2 expression in the stromal cells increased gradually during malignant progression, The results suggest that increased COX-2 expression may be associated with the chemically induced carcinogenic progression of hamster buccal pouch model, The gradual increasing COX-2 expression de tected during the progressive manner toward more malignant lesions shows that the COX-2 protein can have an important role in both the early and the later stages of multistep oral carcinogenesis
본 연구는 포유류인 통로의 존재 여부를 확인하기 위하여 수행되었다. 통로는 비흥분성 세포에서 용적변화와 pH 조절, 이온운반계 등에 중요한 역할을 수행하므로 활발한 세포분화가 이루어지는 난자의 생존과 기능에 필수적 통로 존재 여부를 확인하고자 하였다. 단일통로 전류를 기록하는 patch clamp 기법을 이용하였다. 세포외(pipette) 용액을 140 mM NaCl로 하고 세포내(bath) 용액을 70 mM, 140 mM, 280 mM NaCl로 바꾸
음압에 의한 세포막 신전으로 열리는 Stretch-activated channel(SAC)은 세포의 부피조적, 세포의 분화, 혈관 긴장도의 조절, 호르몬 분비 조절에서 SAC 존재 유무를 확인하기 위하여 patch clamp기법을 시행하여 SAC의 조절기전과 전기생리학적인 성질을 조사하였다. 음압이 주어지기 전에는 관찰되지 않던 단일통로 전류가 -20 cm이하의 음압이 주어졌을 때 관찰되었다. 음압에 의해 열리는 단일통로 전류는 이나 과 같은 일가 양이
세로토닌 수용체는 세로토닌과 반응하여 세포막의 G단백질을 통해 중개단백질 (adenylyl cyclase, phospholipase C, cGMP phosphodiesterase, ion channel)을 활성화시켜, 이뇨, 기억, 발생 등의 다양한 생리적 반응에 관여한다. 곤충세포인 Schneider2 (S2)와 척추동물 세포인 Chinese hamster ovary (CHO)-Kl에서 Aedes 5-HT 수용체 유전자 발현을 비교하기 위해, Aedes 5-HT 수용체 유전자를 형질이입시켰다. 선발된 세포주들(Tr-S2, Tr-CHO)에서 세로토닌 수용체 유전자의 발현은 reverse transcription-PCR, Western blot, immunocytochemistry를 이용하여 확인하였다. 세로토닌 농도증가에 대한 Aedes 5-HT수용체의 기능을 세포 내 cAMP수준을 통해 조사한 결과,Tr-CHO 세포주는 Tr-S2 세포주보다 9배 이상 cAMP수준이 높게 나타났으며, 농도에 의존적이었다. 이 결과는 수용체 유전자가 세포에서 발현되었으나, 세포의 종류와 세포막에 존재하는 G단백질 차이에 따라 중개단백질 활성 차이가 있다는 것을 보여주었다. CHO-Kl 세포에서 Aedes 5-HT 수용체의 기능이 S2 세포보다 더 효율적이며, Aedes 5-HT 수용체를 발현하는 Tr-CHO 세포주는 동력제 또는 대립제 검정에 활용될 수 있을 것으로 기대된다. 것으로 기대된다.
Nitric oxide (NO) plays a key role in the processes of inflammation and carcinogenesis. Three isoforms of NO 야mthase have been identified: endothelial 띠띠c oxide 와nth앓e (NOS), neuronal NOS, and inducible NOS (이OS). The purpose of this study was to investigate the characteristics of iNOS expression in 7, 12-dimethylbenz[alanthracene (DMBA)-induced hamster buccal pouch carcinogenesis. Sixty three outbred young (6-week-old) male Syrian golden hamsters were randomly divided into three groups: DMBA treated group (n=57) and non-treated (n=3), and mineral-oil treated group (n=3). No iNOS activity could be detected in the untreated or mineral oil-treated pouches. 80th cytoplasmic and nuclear stainings were observed in the DMBA-treated pouch kera띠lCX까es. There were iNOS expression 외so in the strorna1 cells. The mean values of iNOS expression in the epithelium increased gradually from control to dysplastic lesions and more to invasive squ따nous cell carcinoma. πle clifference between iNOS expr'않sion in the normal and that the dysplastic and carcinomatous lesions is statistically significant. The mean values of iNOS expression in the stroma increased gradually from control to dysplastic lesions and more to invasive squamous cell carcinoma. The difference between iNOS expression in the normal and that the carcinomatous lesions is statistica11y si맑, ificant. In conclusion, this study has demonstrated that iNOS is expressed in DMBA-induced hamster pouch carcinomas. πlis finding suggests that iNOS expression may be associated with the development of chemically induced oral carcmomas.
As an preliminary experiment for making transgenic animals producing human follicle stimulating hormone (hFSH), we tried to express recombinant hFSH gene in vitro. hFSH is a heterodimeric glycoprotein hormone produced in the anterior pituitary gland. The hormone is essential in the regulation of reproductive processes, such as follicular development and ovulation. Genes encoding the common gonadotrophin alpha subunit and FSH-specific beta subunit were inserted into retroviral vectors under the control of the rat beta actin promoter. Gene transfer to the Chinese hamster ovary (CHO) cells was done by infection of the retroviruses harvested from PT67 packaging cells transfected with recombinant retrovirus vector DNA. After selection with G4l8, PCR and RT-PCR analyses of the G4l8-resistant CHO cells showed successful transfer and expression of both and fragments of the FSH gene.
This study was carried out to evaluate the irritant potential of P-toothpaste in hamster cheek pouch. The test materials were applied once at the beginning of this study into right pouches of hamsters and maintained for 14 days. Animals were administered with P-toothpaste, Bamboo salt toothpaste, D.W. and control solution, respectively. In order to evaluate the irritant potential in mucosa of hamster cheek pouch, we observed clinical signs, mortality, body weights and gross and histopathological findings for 14 days. In all groups, there were neither dead animals nor significant changes of body weights. In addition, there were no differences between D.W. and Ptoothpaste treated group in gross and histopathological findings. Therefore, these results suggest that there was little irritant potential of P-toothpaste in hamster cheek pouch.
흰쥐의 뇌로부터 읽는구조 전체를 포함하는 1.8kbp 크기인 IP_3K를 암호화 하는 IP_3KcDNA 유전자속의 NotⅠ부위 GCGGCCGC를 GCAGCCGC로 site-directed mutagenesis 하여 얻은 변이 IP_3KcDNA를 pSP72·Not2 운반체의 EcoR I 부위에 클로닝하여 증폭시키고, 유전자 재조합 과정을 거쳐서 포유동물의 발현 운반체인 pZIP·NeoSV(X)의 NotⅠ부위에 IP_3KcDNA를 서브클론하였다. 이것을 CCL39 hamster lung fibroblasts 세포에 transfection 하여 발현시키고, anti-IP_3K antibody를 사용하여 Western blot 법으로 효소량과 활성도를 각각 측정한 결과, 효소량은 5배, 효소의 활성도는 무려 16배로서 기대하였던 것보다 훨씬 발현효율이 높았다. 또한 흰쥐의 각 조직속에 IP_3K의 분포와 생화학적인 특성이 논의되었다.
Frozen storage of the oocytes has been used in a few mammalian species including mouse, hamster, human and cattle. However, frozen4hawed oocvtes show different sperm penetration on the levels of the zona pellucida and the plasma memhrane when compared with fresh oocytes. To elucidate biological changes occurring during freezing and thawing, we examined the kinetics of sperm penetration into frozen-thawed hamster oocytes. Oocytes obtained from superovulated female golden hamsters were frozen-thawed in an autofreezer according to an established method. Fresh and frozen4hawed oocytes were fertilized in vitro with capacitated hamster spermatozoa after removing the zona pellucida. The oocytes were examined at 1, 2, 3 and 6 h postinsemination. Sperm penetration found to be 1 h delayed in frozen-thawed oocytes. Other parameters such as degree of polyspermy and decondensing sperm heads were not affected by freezing and thawing. The results suggest that freezing and thawing may cause changes in the egg membrane surface and subsequently which leads to delay in the sperm-egg fusion.