• Animal Bioscience
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• 제36권6호
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• pp.973-979
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• 2023

Objective: The clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (Cas9) system, which is the most efficient and reliable tool for precisely targeted modification of the genome of living cells, has generated considerable excitement for industrial applications as well as scientific research. In this study, we developed a gene-editing and detection system for chick embryo sexing during the embryonic stage. Methods: By combining the CRISPR/Cas9 technical platform and germ cell-mediated germline transmission, we not only generated Z chromosome-targeted knockin chickens but also developed a detection system for fluorescence-positive male chicks in the embryonic stage. Results: We targeted a green fluorescent protein (GFP) transgene into a specific locus on the Z chromosome of chicken primordial germ cells (PGCs), resulting in the production of Z^GFP-knockin chickens. By mating Z^GFP-knockin females (Z^GFP/W) with wild males (Z/Z) and using a GFP detection system, we could identify chick sex, as the GFP transgene was expressed on the Z chromosome only in male offspring (Z^GFP/Z) even before hatching. Conclusion: Our results demonstrate that the CRISPR/Cas9 technical platform with chicken PGCs facilitates the production of specific genome-edited chickens for basic research as well as practical applications.

• 대한방사선기술학회지:방사선기술과학
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• 제46권5호
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• pp.379-394
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• 2023

Targeted Alpha Therapy (TAT) is a new method of cancer treatment that protects normal tissues while selectively killing tumor cells using high cytotoxicity and short range of alpha particles, and target alpha therapy is a highly specific and effective cancer treatment strategy, and its potential has been proven through many clinical and experimental studies. This treatment method accurately delivers alpha particles by selecting specific molecules present in cancer tissue, which has an effective destruction and tumor suppression effect on cancer cells, and one of the main advantages of target alpha treatment is the physical properties of alpha particles. Alpha particles have a very high energy and short effective distance, interacting with target molecules in cancer tissues and having a fatal effect on cancer cells, which is known to cause DNA damage and cell death in cancer cells. TAT has shown positive results in preclinical and clinical studies for various types of cancers, especially those that resist or are unresponsive to existing treatments, but there are several challenges and limitations to overcome for successful clinical transition and application. These include the provision and production of suitable alpha radioisotopes, optimization of target vectors and delivery formulations, understanding and regulation of radiological effects, accurate dosage calculation and toxicity assessment. Future research should focus on developing new or improved isotopes, target vectors, transfer formulations, radiobiological models, combination strategies, imaging techniques, etc. for TAT. In addition, TAT has the potential to improve the quality of life and survival of cancer patients due to the possibility of a new treatment for overcoming cancer, and to this end, prospective research on more carcinomas and more diverse patient groups is needed.

• 대한유전성대사질환학회지
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• 제17권3호
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• pp.96-102
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• 2017

당원병 IX형은 phosphorylase kinase 효소 결핍으로 분해되지 않은 당원이 간 또는 근육에 축적되는 유전성대사이상질환이다. 당원병 IXa형은 당원병 IX형 중 가장 흔한 형태로 PHKA2 유전자 변이로 발생한다. 당원병 IXa형의 임상증상은 간 비대, 간 효소 수치 상승, 성장 지연, 저혈당 등이 있다. 그러나, 이러한 임상 증상은 다른 타입의 당원병의 증상과 비슷하거나 겹쳐서 임상적으로는 구분하기가 어렵다. 저자들은 표적 엑솜 시퀀싱으로 진단된 가족성 당원병 IXa형 증례를 보고하고자 한다. 4세 남아가 간 비대와 간 효소 수치 상승을 주소로 내원하였다. 간 조직검사결과 간세포에 당원이 축적되어 있어 당원병을 의심하였으나 G6PC 유전자 검사는 음성이었다. 이에 당원병 타입을 감별진단 하기 위해 표적 엑솜시퀀싱을 시행하였으며, PHKA2 유전자에서 질환과의 연관성이 이미 보고된 바 있는 c.3632C>T (p.Thr121Met) 변이가 반접합체(hemizygote)로 발견되어 당원병 IXa로 진단하였다. 가족 유전자 검사를 통해 어머니가 이형접합체 보인자임을 확인하였으며, 남동생이 같은 변이를 가진 반접합체임을 확인하였다. 28개월 된 환자의 남동생 역시 신체 검진 상 간 비대가 있었으며, 혈액검사상 간 효소 수치가 상승되어 있어 같은 질환으로 확진하였다. 이환된 형제 모두 생 옥수수 전분 섭취와 복합 탄수화물을 섭취하도록 식이 조절을 하였으며 2년 추적관찰 동안 정상 성장 발달을 보이고 있다. 당원병과 같이 임상적으로 구분이 어려우며 유전학적으로 다양한 유전자 변이를 보이는 당원병과 같은 질환의 분자 유전학적 감별진단에 표적 엑솜 시퀀싱이 유용한 진단법이 될 수 있다. 신속하고 정확한 분자 유전학적 감별진단을 통해 환자와 보호자에게 질병의 적절한 치료법, 질병의 예후에 관한 정확한 정보를 제공할 수 있을 뿐 아니라, 적절한 유전상담을 제공할 수 있다.

• Asian Pacific Journal of Cancer Prevention
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• 제16권2호
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• pp.647-652
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• 2015

Background: Lung cancer was one of the most common cancers in both men and women all over the world. In this study, we aimed to clarify who could survive after long term chemotherapy in patients with advanced non-small cell lung cancer (NSCLC). Methods: We enrolled 186 patients with stage IV NSCLC after long term chemotherapy from Jun 2006 to Nov 2014 diagnosed in Jiangsu Cancer Hospital. Multiple variables like age, gender, smoking, histology of adenocarcinoma and squamous-cell cancer, number of metastatic sites, metastatic sites (e.g. lung, brain, bone, liver and pleura), hemoglobin, lymphocyte rate (LYR), Change of LYR during multiple therapies, hypertension, diabetes, chronic bronchitis, treatments (e.g.radiotherapy and targeted therapy) were selected. For consideration of factors influencing survival and response for patients with advanced NSCLC, logistic regression analysis and Cox regression analysis were used in an attempt to develop a screening module for patients with elevated survival after long term chemotherapy become possible. Results: Of the total of 186 patients enrolled, 69 survived less than 1 year (short-term group), 45 one to two years, and 72 longer than 3 years (long-term group). For logistic regression analysis, the short-term group was taken as control group and the long-term group as the case group. We found that age, histology of adenocarcinoma, metastatic site (e.g. lung and liver), treatments (e.g. targeted therapy and radiotherapy), LYR, a decreasing tendency of LYR and chronic bronchitis were individually associated with overall survival by Cox regression analysis. A multivariable Cox regression model showed that metastatic site (e.g. lung and liver), histology of adenocarcinoma, treatments (e.g. targeted therapy and radiotherapy) and chronic bronchitis were associated with overall survival. Thus metastatic site (e.g. lung and liver) and chronic bronchitis may be important risk factors for patients with advanced NSCLC. Gender, metastatic site (e.g. lung and liver), LYR and the decreasing tendency of LYR were significantly associated with long-term survival in the individual-variable logistic regression model (P<0.05). On multivariate logistic regression analysis, gender, metastatic site (e.g. lung and liver) and the decreasing tendency of LYR associated with long-term survival. Conclusions: In conclusion, female patients with stage IV adenocarcinoma of NSCLC who had decreasing tendency of LYR during the course therapy and had accepted multiple therapies e.g. more than third-line chemotherapy, radiotherapy and/or targeted therapy might be expected to live longer.

• Asian-Australasian Journal of Animal Sciences
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• 제24권2호
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• pp.222-229
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• 2011

This study was conducted to determine the effects of dietary multi-nutritional targeted supplementation according to different growth stages on performance, blood metabolites and carcass characteristics of Korean native Hanwoo steers. Thirty two Hanwoo steers, 6 months of age and weighing $159.2{\pm}24.1\;kg$, were distributed into 2 groups: control group without any supplements, and multi-nutritional targeted supplementation (MNTS) treatment group supplied with wheat bran and catechins (WBC, 8 to 16 months of age), ruminally protected amino acid-enriched fatty acid (RPAFA, 17 to 28 months of age) and ruminally protected choline with vitamin E (RPCV, 22 to 28 months of age), respectively. Average daily gain was not significantly different between the two groups. During the late fattening period, concentrate, rice straw and dry matter intakes were relatively lower in MNTS than in the control group. Rib eye area, back fat thickness and yield index were similar between the two groups. Meat color was relatively lower in MNTS compared with the control group. The appearance rate of high quality grade ($1^{++}$, $1^+$ and 1) tended to be higher in MNTS compared with the control group. Thus, the present results indicate that dietary multi-nutritional targeted supplementation at different growth stages could be recommendable to increase income according to production of high quality Hanwoo beef without any negative effects on growth performance and carcass characteristics.

• 한국전산구조공학회논문집
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• 제30권5호
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• pp.371-380
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• 2017

미국의 내진설계기준인 ASCE/SEI 7-10은 구조물 붕괴성능에 대한 불확실성을 고려하지 않는 등재해도 기반 내진설계의 문제점을 해결하기 위해 위험도 기반 내진설계 개념을 도입하였다. 하지만 현행 국내 내진설계기준의 경우 한반도 내에서 발생한 큰 규모의 지진기록과 구조물의 붕괴성능과 관련된 연구의 부족으로 위험도 기반 내진설계 개념을 반영하지 않고 있다. 본 연구에서는 철골 보통중심가새골조를 표본건물로 선정하여 위험도 기반 내진성능평가를 수행하였다. 건물이 위치한 지역, 높이, 지반조건을 변수로 바탕으로 표본건물에 대한 붕괴성능 평가를 수행하였으며, 국내 지진기록의 특성을 반영할 수 있는 경험적 스펙트럴 형상 예측 모델을 활용하여 지진재해도 곡선을 작성하였다. 이를 활용하여 국내 주요 도시에 위치한 철골 보통중심가새골조의 붕괴확률을 위험도 적분 개념에 따라 평가하였다. 국내 주요 도시에 위치한 철골 보통중심가새골조의 붕괴확률을 평가한 결과, 현행 건축구조기준에 따라 설계된 표본건물은 본 연구에서 고려한 해석 변수에 따라 붕괴확률에 상당한 차이를 보였다. 특히 국내 건축구조기준의 경우 철골 보통중심가새골조에 대한 높이제한이 없어 일부 고층표본건물에서 목표 위험도인 50년간 1%의 붕괴확률을 초과하는 것으로 평가되었다.

• 한국수정란이식학회지
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• 제29권4호
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• pp.339-344
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• 2014

The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR associated protein (Cas9) system can be applied to produce transgenic pigs. Therefore, we applied CRISPR/Cas9 system to generate FoxN1-targeted pig parthenogenetic embryos. Using single guided RNA targeted to pig FoxN1 genes was injected into cytoplasm of in vitro matured oocyte before electrical activation. In results, regardless of the concentrations of vector, the cleavage rate were significantly (p<0.05) decreased ($4ng/{\mu}l$, 51.24%; $8ng/{\mu}l$, 40.88%; and $16ng/{\mu}l$; 45.22%) compared to no injection group (70.44%). The blastocyst formation rates were also decreased in vector injected 3 groups ($4ng/{\mu}l$, 7.96%; $8ng/{\mu}l$, 6.4%; and $16ng/{\mu}l$; 9.04%) compared to no injection group (29.07%). In addition, the blastocyst formation rates between sham injected group (13.51%) and no injection group (29.07%) also showed significant difference (p<0.05). The mutation rates were comparable between groups ($4ng/{\mu}l$, 18.4%; $8ng/{\mu}l$, 12.5%; and $16ng/{\mu}l$; 20.0%). The sequencing analysis showed that blastocysts derived from each group were successfully mutated in FoxN1 loci regardless of the vector concentrations. However, the deletion patterns were higher than the patterns of point mutation and insertion regardless of the vector concentrations. In conclusion, we described that cytoplasmic microinjection of FoxN1-targeted CRISPR/Cas9 vector could efficiently generate transgenic pig parthenogenetic embryos in one-step.

• 미생물학회지
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• 제33권4호
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• pp.257-261
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• 1997

겨울철 소양호에서 세균 군집 구조를 파악하고자 총세균수와 EUB338, ALF1b, BET4a, GAM42a와 CF probe 등 fluorescent rRNA targeted oligonucleotide probe 와 반응하는 세균 개체수를 수심별로 측정하였다. 총세균수는 $0.7{\times}10^6{\sim}1.1{\times}10^6cell{\cdot}ml^{-1}$이였으며, 5 m와 10 m 수층에서 높게 나타났다. 총세균수에 대한 Eubacteria의 비율은 34~90%이였으며, 5 m와 10 m에서 낮게 나타났다. Proteobacteria ${\alpha}$-group은 Eubacteria의 10.8-28.7%, ${\beta}$-group은 4.5-53.5%, ${\gamma}$-group은 4.9-35.5%, 그리고 Cytophaga-Flavobacterium group은 6.1-21.1%이였다. 0-5 m 수심에서는 ${\beta}$-group이 28.6-53.5%로 우점하고 있었으며, 10 m에서는 ${\gamma}$-group이 35.5%로 우점하였다. 30, 50 m 수심에서는 ${\alpha}$-group과 Cytophaga-Flavobacterium group이 우점하였다. 세균 군집 구조로 보면 0-2 m, 5-10 m 그리고 30-50 m의 3개층은 각각 독특한 특징을 나타내었다. 이 방법으로 호수 생태계에 대한 새로운 정보를 얻을 수 있었다.

• Journal of Microbiology and Biotechnology
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• 제31권7호
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• pp.933-941
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• 2021

Ginsenoside Rg4 is a rare ginsenoside that is naturally found in ginseng, and exhibits a wide range of biological activities including antioxidant and anti-inflammatory properties in several cell types. The purpose of this study was to use an in vivo model of hair follicle (HF)-mimic based on a human dermal papilla (DP) spheroid system prepared by three-dimensional (3D) culture and to investigate the effect of Rg4 on the hair-inductive properties of DP cells. Treatment of the DP spheroids with Rg4 (20 to 50 ㎍/ml) significantly increased the viability and size of the DP spheres in a dose-dependent manner. Rg4 also increased the mRNA and protein expression of DP signature genes that are related to hair growth including ALP, BMP2, and VCAN in the DP spheres. Analysis of the signaling molecules and luciferase reporter assays further revealed that Rg4 induces the activation of phosphoinositide 3-kinase (PI3K)/AKT and the inhibitory phosphorylation of GSK3β, which activates the WNT/β-catenin signaling pathway. These results correlated with not only the increased nuclear translocation of β-catenin following the treatment of the DP spheres with Rg4 but also the significant elevation of mRNA expression of the downstream target genes of the WNT/β-catenin pathway including WNT5A, β-catenin, and LEF1. In conclusion, these results demonstrated that ginsenoside Rg4 promotes the hair-inductive properties of DP cells by activating the AKT/GSK3β/β-catenin signaling pathway in DP spheres, suggesting that Rg4 could be a potential natural therapy for hair growth.

• Asian Pacific Journal of Cancer Prevention
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• 제17권5호
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• pp.2727-2728
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• 2016