• 한국발생생물학회지:발생과생식
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• 제4권1호
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• pp.7-12
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• 2000

수정에 의한 배 발생은 정자가 난자 내로 침입하여 정자와 난자의 반수체 핵질이 융합되고 이어 유사분열로 이어지는 과정에서 시작된다. 하지만 수정 및 초기 배 발생 동안 자웅 핵질과 난 세포질 구성 요소 상호간의 작용기전에 관해서는 명확히 알려져 있지 않은 부분이 많다. 수정보조기법인 세포질 내 정자 직접 주입법의 개발은 남성불임치료에 혁신적인 기술로 자리잡고 있을 뿐만 아니라 포유동물의 수정과정을 이해하는데 많은 도움을 주고 있다. 핵치환에 의한 복제동물 생산기법도 분화된 핵이 난 세포질 내에서 재 분화 (reprogramming)하여 발생하는 유일한 과정으로 세포질 구성요소들의 상호작용과 발생 조절 기능을 이해하는데 도움을 준다. 최근 몇 년간 돼지 난자 세포질에 정자 및 원형정자 직접주입, 세포질 이식, 세포질 융합 및 핵치환 한 후 난자의 발생과정을 간접 면역형광 분석법과 주사 전자현미경으로 조사하였다. 이러한 연구를 통해 체외수정, 세포질 이식 및 정자직접 주입법 등과 같은 임상치료기술 과 핵치환에 의한 복제동물생산 기법의 개선에 필요한 기초자료를 얻을 수 있었고, 포유동물 난자의 후생적 발생과정 (epigenesis)에 관해 공부할 수 있었다.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2000년도 국제심포지움
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• pp.1-2
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• 2000

High production of milk and its components are necessary to allow maximal growth of developing piglets. In this study, transgenic pigs were produced containing the $\alpha$-lactalbumin gene, whose product is a potential limiting component in the production of milk. Two lines of transgenic pigs were produced to analyze the effects that overproduction of the milk protein $\alpha$-lactalbumin may have on milk production and piglet growth. Transgenic pigs were produced through microinjection of the bovine $\alpha$-lactalbumin gene. The gene construct contained 2.0 kb of 5 flanking region, the 2.0 kb coding region and 329 bp of 3 flanking region. Sows hemizygous for the transgene produced as much as 0.9 g of bovine $\alpha$-lactalbumin per liter of pig milk. The production of the bovine protein caused approximately a 50 % increase in the total $\alpha$-lactalbumin concentration in pig milk throughout lactation. The concentration of bovine $\alpha$-lactalbumin was highest on day 0 and 5 of lactation and decreased as lactation progressed. The ratio of bovine to porcine $\alpha$-lactalbumin changed during the sow's lactation. This ratio was 4.3 to 1 on day 0 of lactation, but by day 20 of lactation the ratio was 0.43 to 1. This suggested that the bovine transgene and the endogenous porcine gene were under slightly different control mechanisms. The higher level of total $\alpha$-lactalbumin present on day 0 of lactation was correlated with higher lactose percentage on day 0 in transgenic sows (3.8 %) as compared to controls (2.6 %) (P < 0.01). Although there was also a trend for higher lactose percentage in transgenic sows on day 5 and 10 of lactation, no significant differences were observed. These data suggest that $\alpha$-lactalbumin is limiting early in lactation of swine. Furthermore, higher concentrations of $\alpha$-lactalbumin early in lactation may boost milk output.

• Archives of Plastic Surgery
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• 제45권5호
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• pp.411-417
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• 2018

Background Accessory auricles (AAs) are common congenital anomalies. We present a new classification according to location and shape, and propose a system for coding the classifications. Methods This study was conducted by reviewing the records of 502 patients who underwent surgery for AA. AAs were classified into three anatomical types: intraauricular, preauricular, and buccal. Intraauricular AAs were divided into three subtypes: intracrural, intratragal, and intralobal. Preauricular AAs were divided into five subtypes: precrural, superior pretragal, middle pretragal, inferior pretragal, and prelobal. Buccal AAs were divided into two subtypes: anterior buccal and posterior buccal. AAs were also classified according to their protrusion pattern above the surrounding surface: pedunculated, sessile, areolar, remnant, and depressed. Pedunculated and sessile AAs were subclassified as spherical, ovoid, lobed, and nodular, according to their body shape. Cartilage root presence and family history of AA were reviewed. A coding system for these classifications was also proposed. Results The total number of AAs in the 502 patients was 1,003. Among the locations, the superior pretragal subtype (27.6%) was the most common. Among the protrusion patterns and shapes, pedunculated ovoid AAs were the most common in the preauricular (27.8%) and buccal areas (28.0%), and sessile lobed AAs were the most common in the intraauricular area (48.7%). The proportion of AAs with a cartilage root was 78.4%, and 11% of patients had a family history. The most common type of preauricular AA was the superior pretragal pedunculated ovoid AA (13.2%) with a cartilage root. Conclusions This new system will serve as a guideline for classifying and coding AAs.

• 한국수정란이식학회지
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• 제26권2호
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• pp.123-128
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• 2011

In vitro maturation and activation of oocytes are primary steps towards biotechnological manipulation in embryology. The objectives of the present study were to determine the oocyte recovery rate per ovary, in vitro maturation rates of oocytes and rates of parthenogenetically activation of matured oocytes in Black Bengal goats. All visible follicles were aspirated to recover follicular fluid from individual ovaries (number of ovaries = 456). The immature cumulus oocyte complexes (COCs; n = 1289) were cultured in tissue culture medium (TCM)-199 supplemented with 10% (v/v) fetal bovine serum (FBS) for 27 hours at $39^{\circ}C$ with 5% $CO_2$ in humidified air. The matured oocytes (n = 248) were activated with 5 ${\mu}M$ ionomycin for 5 minutes followed by treatment with 2 mM 6-dimethylaminopurine (6-DMAP) for 4 hours. After activation, oocytes were cultured for another 14 hours in TCM-199 supplemented with bovine serum albumin (BSA) at $39^{\circ}C$ with 5% $CO_2$ in humidified air. The pronucleus formation in activated oocytes was determined by staining with 1% orcein (whole mount technique). Matured oocytes (n = 176) without activation stimuli were used as control. The mean number of oocytes recovered per ovary was $3.5{\pm}0.5$. The proportion of oocytes matured in vitro, confirmed by the presence of first polar body, was $42.1{\pm}4.7%$. Parthenogenetic activation, evidenced by formation of pronucleus, occurred in $37.2{\pm}15.8%$ of matured oocytes. No pronucleus formation was observed in control oocytes. In conclusion, a combination of ionomycin and 6-DMAP induces activation in one third of Black Bengal goats' oocytes.

• Asian Pacific Journal of Cancer Prevention
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• 제15권16호
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• pp.6955-6960
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• 2014

Background: The cytokinesis-block micronucleus (CBMN) assay is a standard cytogenetic tool employed to evaluate chromosomal damage subsequent to pesticide exposure. Objectives: To evaluate the pooled levels of total micronuclei (MN) and binucleated cells with micronuclei (MNC) in 1000 binucleated lymphocytes among population occupationally exposed to pesticides and further determine the more sensitive biomarker of CBMN. Materials and Methods: A meta-analysis on the pooled levels of MN and MNC in binucleated lymphocytes among occupationally pesticide-exposed populations was conducted using STATA 10.0 software and Review Manager 5.0.24 in this study. Results: We found significant differences in frequencies of MN and MNC in 1000 binucleated lymphocytes between pesticide-exposed groups and controls, and the summary estimates of weighted mean difference were 6.82 [95% confidence interval (95% CI): 4.86-8.78] and 5.08 (95% CI: 2.93-7.23), respectively. However, when we conducted sensitivity analyses further, only the MN remained statistically different, but not the MNC, the summary estimates of weight mean difference were 2.86 (95% CI: 2.51-3.21) and 0.50 (95% CI: -0.16-1.17), respectively. We also observed pesticide-exposed subjects had significantly higher MN frequencies than controls among smokers and nonsmokers, male and female populations, and American, Asian and European countries in stratified analyses. Conclusions: The frequency of MN in peripheral blood lymphocytes might be a more sensitive indicator of early genetic effects than MNC using the CBMN assay for occupationally pesticide-exposed populations.

• Asian Pacific Journal of Cancer Prevention
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• 제17권11호
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• pp.4959-4963
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• 2016

Objective:Cervical cancer is one of the major causes of mortality of Uyghur women in Xinjiang, China. Although increased expression of orexin receptor (OxR), known to be strongly expressed in human placenta, has a proven relation to some cancers, there have been few studies of cervical cancer. Thus, we explored this question by evaluating the expression of orexin receptor as a biomarker for screening early stage of cervical cancer in Uyghur women with highest occurrence rate of cervical cancer in China. Study Design: We used polymerase chain reaction (PCR) and immunohistochemical staining to determine the expression of both Ox1R and Ox2R in cervical cancer and cervicitis biopsies collected from Uyghur women infected with human papilloma virus (HPV)16. The expression rate was compared between cervical cancers of low, intermediate and high differentiation and cervicitis. Results:Although there was no significant difference in the expression rate of Ox1R between groups, Ox2R was significantly overexpressed in cervical cancer patients when compared to the cervicitis group. Ox1R was negative in normal human placenta while Ox2R was positive. Conclusions: While expression of Ox1R had no correlation with invasion or metastatic potential, Ox2R demonstrated elevation in cervical cancer with heterogeneity in groups with different metastatic potential, in the human placenta as well, implying that it might serve as an indicator of invasive capacity along with other indices.

• Journal of Korean Neurosurgical Society
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• 제57권5호
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• pp.335-341
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• 2015

Objective : The main causes of spinal cord ischemia are a variety of vascular pathologies causing acute arterial occlusions. We investigated neuro-protective effects of kefir on spinal cord ischemia injury in rats. Methods : Rats were divided into three groups : 1) sham operated control rats; 2) spinal cord ischemia group fed on a standard diet without kefir pretreatment; and 3) spinal cord ischemia group fed on a standard diet plus kefir. Spinal cord ischemia was performed by the infrarenal aorta cross-clamping model. The spinal cord was removed after the procedure. The biochemical and histopathological changes were observed within the samples. Functional assessment was performed for neurological deficit scores. Results : The kefir group was compared with the ischemia group, a significant decrease in malondialdehyde levels was observed (p<0.05). Catalase and superoxide dismutase levels of the kefir group were significantly higher than ischemia group (p<0.05). In histopathological samples, the kefir group is compared with ischemia group, there was a significant decrease in numbers of dead and degenerated neurons (p<0.05). In immunohistochemical staining, hipoxia-inducible factor-$1{\alpha}$ and caspase 3 immunopositive neurons were significantly decreased in kefir group compared with ischemia group (p<0.05). The neurological deficit scores of kefir group were significantly higher than ischemia group at 24 h (p<0.05). Conclusion : Our study revealed that kefir pretreatment in spinal cord ischemia/reperfusion reduced oxidative stress and neuronal degeneration as a neuroprotective agent. Ultrastructural studies are required in order for kefir to be developed as a promising therapeutic agent to be utilized for human spinal cord ischemia in the future.

• 대한수의학회지
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• 제57권2호
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• pp.89-95
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• 2017

This study was conducted to determine the effects of biophoton treatment during in vitro maturation (IVM) and/or in vitro culture (IVC) on oocyte maturation and embryonic development in pigs. An apparatus capable of generating homogeneous biophoton energy emissions was placed in an incubator. Initially, immature pig oocytes were matured in the biophoton-equipped incubator in medium 199 supplemented with cysteine, epidermal growth factor, insulin, and gonadotrophic hormones for 22 h, after which they were matured in hormone-free medium for an additional 22 hr. Next, IVM oocytes were induced for parthenogenesis (PA) or provided as cytoplasts for somatic cell nuclear transfer (SCNT). Treatment of oocytes with biophoton energy during IVM did not improve cumulus cell expansion, nuclear maturation, intraoocyte glutathione content, or mitochondrial distribution of oocytes. However, biophoton-treated oocytes showed higher (p < 0.05) blastocyst formation after PA than that in untreated oocytes (50.7% vs. 42.7%). In an additional experiment, SCNT embryos produced from biophoton-treated oocytes showed a greater (p < 0.05) number of cells in blastocysts (52.6 vs. 43.9) than that in untreated oocytes. Taken together, our results demonstrate that biophoton treatment during IVM improves developmental competence of PA- and SCNT-derived embryos.

• Asian Pacific Journal of Cancer Prevention
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• 제13권8호
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• pp.4101-4105
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• 2012

Purpose: The degree of radiation injury to kidneys which are located within the limits of radiotherapy area is determined by the volume and the dose of radiation to which the organ is exposed. When the tolerance dose of the kidney is exceeded after a latent period of 6 months acute nephritis develops and after 18 months chronic nephritis ensues. Melatonin is known to prevent the oxidative injury of toxins and radiotherapy with its free radical scavenging capacity. Methods and Materials: In this study 8 weeks old 24 Sprague -Dawley rats were allocated into 4 groups: Control group; Radiotherapy group (20 Gy bilaterally in 5 fractions); Melatonin group (10 mg/kg intraperitoneally), and Melatonin+radiotherapy group (20 Gy Radiotherapy in 5 fractions+ melatonin 10 mg/kg intraperitoneally). After a follow-up period of 6 months BUN was determined in all groups. After rats were euthanized the kidneys were removed for histopathological examination under both light and electron microscopes. Results: After 6 months follow-up, both at light and electron microscopy levels, the rats in radiotherapy+melatonin group were significantly protected against the radiation injury comparing to radiotherapy group (p<0.05). Conclusion: It was shown in this experimental model that melatonin has protective effects against radiation injury to kidneys.

• Asian Pacific Journal of Cancer Prevention
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• 제16권10호
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• pp.4199-4202
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• 2015

Background: Hepatitis B virus infection is one of the major world health problems. Epigallocatechin-3 gallate is the major component of the polyphenolic fraction of green tea and it has an anti-viral, anti-mutagenic, anti-tumorigenic, anti-angiogenic, anti-proliferative, and/or pro-apoptotic effects on mammalian cells. In this study, our aim was to investigate the inhibition of HBV replication by epigallocatechin-3 gallate in the Hep3B2.1-7 hepatocellular carcinoma cell line. Materials and Methods: HBV-replicating Hep3B2.1-7 cells were used to investigate the preventive effects of epigallocatechin-3 gallate on HBV DNA replication. The expression levels of HBsAg and HBeAg were determined using ELISA. Quantitative real-time-PCR was applied for the determination of the expression level of HBV DNA. Results: Cytotoxicity of epigallocathechin-3-gallate was not observed in the hepatic carcinoma cell line when the dose was lower than $100{\mu}M$. The ELISA method demonstrated that epigallocatechin-3 gallate have strong effects on HBsAg and HBeAg levels. Also it was detected by real-time PCR that epigallocatechin-3 gallate could prevent HBV DNA replication. Conclusions: The obtained data pointed out that although the exact mechanism of HBV DNA replication and related diseases remains unclear, epigallocatechin-3 gallate has a potential as an effective anti-HBV agent with low toxicity.