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Mutational and Functional Analysis of HPV-16 URR Derived from Korean Cervical Neoplasia
Park, Jong-Sub,Hwang, Eun-Seong,Lee, Chan-Jae,Kim, Chan-Joo,Rha, Jong-Gu,Kim, Weung-Jo,Namkoong, 0Sung-Eun,Um, Soo-Jong 가톨릭대학교 2000 Bulletin of The Catholic Research Institutes of Me Vol.28 No.-
Objective : The YY1 mutation has been suggested as one of the indicators that explains development of cervical neoplasia by episomal-type HPV. To extend this hypothesis, we examined whether a mutations(s) in the YY1 site is functionally related to the invasiveness of cervical neoplasia and the physical status of HPV DNA. Methods : The URR sequences were obtained by PCR amplification of HPV-16 genome from CIN and invasive cancer patients and cloned into pUC18 for sequencing and into pBLCAT8+ for functional CAT assay. Results : Our previous data classified HPV-infected patients into three groups : 3 cancer cases carrying episomal HPV DNA ; 12 cancer cases carrying integrated HPV DNA; 12 CIN cases carrying episomal HPV DNA. The specific variants in HPV-16 URR were found in Korean women : G→A transition at nt 7520 (200%, 27/27), A→C transition at nt 7729 (70% ; 21/27). Selective mutations were observed at the YY1 binding sites of HPV-16 URR in the 3 patients with invasive cervical cancer who have the episomal forms of HPV-16 DNA : A→C transition at nt 7484 and G→A transition at nt 7488(YY1-binding site 2 ; from 7481 to 7489). Additionally, C→T transition at nt 7785(YY1-binding site 3 ; from 7781 to 7790) was transition at nt 7785(YY1-binding site 3 ; from 7781 to 7790) was found in 2 of 3 patients. No YY1 site mutations were detected in the 12 CIN patients and in the HPV-integrated invasive cancer patients. To determine whether these mutations have effects on the expression of HPV E6/E7 genes driven by URR, the transient transfection assay was employed using URR-CAT reporter plasmid. The relative activities of three URR mutants from episomal HPV-16 DNA of cervical cancers were two-to fourfold higher than that of the HPV-16 URR prototype. In contrast, the URRs from integrated HPV-16 DNA in cervical cancer and from episomal HPV-16 DNA in CIN, where no mutation of the YY1 binding site was detected, showed similar levels of promoter activity to that of the URR prototype. Conclusions : Our results support the hypothesis that the mutation the YY1 binding site is functionally related to the development of cervical neoplasia caused by episomal HPV-16 DNA in Korean cervical cancer patients. Thus, mutation in the YY1 site of episomal HPV-16 URR may play a corresponding role of HPV integration in the progression of progression of cervical cancer. (Gynecologic Oncology 74:23-29, 1999)
Yi, Chong Seong,Rha, Jong Gu,Lee, Jong Kun,Kim, Yong Wook,Shin, Jong Chul,Kim, Soo Pyung THE CATHOLIC UNIVERSITY OF KOREA 1997 Bulletin of The Catholic Research Institutes of Me Vol.25 No.-
Respiratory distress syndrome(RDS) is a major cause of death in premature neonates, and it is caused by the failure of morphological and biochemical lung maturation(synthesis and secretion of lung surfactant). It is known that cortisol, thyroxine, prolactin, epidermal growth factor(EGF), and estrogen accelerate the lung maturation. Cortisol and thyroxine are currently used in the antenatal treatment for the prevention of RDS in premature neonates. In order to evaluate the effect of EGF on the levels of cortsol, thyroxine, and prolactin, this study was undertaken. Phosphate buffered saline(PBS) with and without EGF was directly injected into the 25 days' gestational fetus in uterus. Blood was collected for the measurement of cortisol, thyroxine, and prolactin one day or two days after the injection. Body weights and lung weights were also measured. The results were as follows : 1. There was no significant difference in body weights and lung weights between PBS-treated group(control group) and EGF-treated group(experimental group), 24 hours and 48 hours after the injection. 2. 24 hours after the injection, the levels of cortisol were significantly increased in the BGF-treated group compared with those in the PBS-treated group. However 48 hours after the injection, there was no significant difference in the levels of cortisol between the two groups. The levels of thyroxine and prolactin in the EGF-treated group did not significantly differ from those in the PBS-treated group 24 hours and 48 hours after the injection. In conclusion, in vivo, the synthesis of cortisol may be affected by EGF treatment, which suggests that the action of EGF for lung maturation may be partially mediated by the increased endogenous levels of cortisol.
Significance of Standard Curve Transformation on the Quality Control of the Radioimmunoassay
Kim, Seung Jo,Rha, Jong Gu CATHOLIC MEDICAL CENTER 1978 Bulletin of the Clinical Research Institute Vol.6 No.1
The radioimmunoassay(RIA) has become a widespread analytic method of various hormones and a potent diagnostic and therapy-monitoring tool. We have measured polypeptide hormones(LH, FSH, hPPL, and hCG β-subunit) with Hypolab kits, progesterone by modified method by Devilla et al., and estradiol by Johanson & Edquvist method. We participated in a part of the 1977 WHO programme for the standardization and quality control of RIA, so polypeptide hormones(LH, FSH, hPPL_ and steroid hormones(estradiol, progesterone, testosterone, contisol) were assayed in accordance with WHO working documents.
Sperm-cervical Mucus Interaction Using Bovine Cervical Mucus and Hen's Egg White
Kim, Jin Hong,Ryu, Ki Sung,Rha, Jong Gu,Kim, Soo Pyung,Lee, Hun Young CATHOLIC MEDICAL CENTER 1990 Bulletin of the Clinical Research Institute Vol.18 No.1
Recently, much attention has been directed to the feasibility of substituting mucus for Human cervical mucus (HCM). Therefore, this study was performed in order to comparison between bovine cervical mucus (BCM) and fraction of hen’s egg white (FEW) as substituting mucus for human cervical mucus with the use of ejaculated human spermatozoa from 15 fertile men (Group 1), 62 infertile couples (Group 2) and 13 male partners (Group 3) with proven male infertility factors of 62 infertile couples. Sperm-cervical mucus interaction rests were performed in a flat capillary tubes with bovine cervical mucus and fraction of egg white. The depth of penetration was observed microscopically. Results were as follows: 1. A significant correlation was found between sperm penetration of Group 2 in BCM and FEW (P < 0.001). 2. The depth of sperm penetration in both BCM and FEW were signifirant1y correlated with sperm motility and progression of seminal parameters (P < 0.001). 3. The mean values of sperm penetration for both bovine cervical mucus and fraction of hen’s egg white were significantly different when sperm penetration of Group 1 and Group 3 were compared (P < 0.001).