• Laboraroty Animal Research
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• v.34 no.4
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• pp.166-175
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• 2018

Recombination activating gene-2 (RAG-2) plays a crucial role in the development of lymphocytes by mediating recombination of T cell receptors and immunoglobulins, and loss of RAG-2 causes severe combined immunodeficiency (SCID) in humans. Rag-2 knockout mice created using homologous recombination in ES cells have served as a valuable immunodeficient platform, but concerns have persisted on the specificity of Rag-2-related phenotypes in these animals due to the limitations associated with the genome engineering method used. To precisely investigate the function of Rag-2, we recently established a new Rag-2 knockout FVB mouse line ($Rag-2^{-/-}$) manifesting lymphopenia by employing a CRISPR/Cas9 system at Center for Mouse Models of Human Disease. In this study, we further characterized their phenotypes focusing on histopathological analysis of lymphoid organs. $Rag-2^{-/-}$ mice showed no abnormality in development compared to their WT littermates for 26 weeks. At necropsy, gross examination revealed significantly smaller spleens and thymuses in $Rag-2^{-/-}$ mice, while histopathological investigation revealed hypoplastic white pulps with intact red pulps in the spleen, severe atrophy of the thymic cortex and disappearance of follicles in lymph nodes. However, no perceivable change was observed in the bone marrow. Moreover, our analyses showed a specific reduction of lymphocytes with a complete loss of mature T cells and B cells in the lymphoid organs, while natural killer cells and splenic megakaryocytes were increased in $Rag-2^{-/-}$ mice. These findings indicate that our $Rag-2^{-/-}$ mice show systemic lymphopenia with the relevant histopathological changes in the lymphoid organs, suggesting them as an improved Rag-2-related immunodeficient model.

• Laboraroty Animal Research
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• v.34 no.4
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• pp.302-310
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• 2018

CD47 (integrin-associated protein), a multi-spanning transmembrane protein expressed in all cells including red blood cells (RBCs) and leukocytes, interacts with signal regulatory protein ${\alpha}$ ($SIRP{\alpha}$) on macrophages and thereby inhibits phagocytosis of RBCs. Recently, we generated a novel C57BL/6J CD47 knockout ($CD47^{-/-}$ hereafter) mouse line by employing a CRISPR/Cas9 system at Center for Mouse Models of Human Disease, and here report their hematological phenotypes. On monitoring their birth and development, $CD47^{-/-}$ mice were born viable with a natural male-to-female sex ratio and normally developed from birth through puberty to adulthood without noticeable changes in growth, food/water intake compared to their age and sex-matched wild-type littermates up to 26 weeks. Hematological analysis revealed a mild but significant reduction of RBC counts and hemoglobin in 16 week-old male $CD47^{-/-}$ mice which were aggravated at the age of 26 weeks with increased reticulocyte counts and mean corpuscular volume (MCV), suggesting hemolytic anemia. Interestingly, anemia in female $CD47^{-/-}$ mice became evident at 26 weeks, but splenomegaly was identified in both genders of $CD47^{-/-}$ mice from the age of 16 weeks, consistent with development of hemolytic anemia. Additionally, helper and cytotoxic T cell populations were considerably reduced in the spleen, but not in thymus, of $CD47^{-/-}$ mice, suggesting a crucial role of CD47 in proliferation of T cells. Collectively, these findings indicate that our $CD47^{-/-}$ mice have progressive hemolytic anemia and splenic depletion of mature T cell populations and therefore may be useful as an in vivo model to study the function of CD47.

• The Korea Journal of Herbology
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• v.37 no.6
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• pp.45-52
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• 2022

Objective : In this study, we conducted to analysis the patents about Korean medicine based treatment technology for pet. Methods : We collected the Korean, Japan, USA, China, EU and PCT patent for the Korean medicine-based treatment technology for pet using WIPSON data base registered by October 4, 2020 were selected. Results : Patents for the development of pet Korean medicine-based treatment technology have been constantly applied since the mid-2000s, starting in 1990, with a total of 84 applications. The number of patent applications filed in Korea was the largest, after Japan and China. The detailed composition of the applications reported until now is pharmaceutical compositions for the treatment of pet were the most applied with 39%(33 cases), followed by feed used as stocks for pet at 36%(30 cases). According to the patent application rate by disease, most of applications rate composed to be associated with immunity in 2009 to 2013 with 40% of applications. In 2014 to 2018 43% of patent applications are associated with gastrointestinal/digestive system. In this regard, multiple applications were filed in the order of immunity, digestive system, obesity and urinary system. Conclusion : This study analyzed the trend of 84 patents about Korean medicine-based treatment technology for pet to date to establish the basis for future research and development. There were many applications related to immunity from 2009 to 2013, and since then, related to digestive disease from 2014 to 2018.

• Journal of Ginseng Research
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• v.47 no.2
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• pp.337-346
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• 2023

Background: Ginsenoside Rb2, a major active component of Panax ginseng, has various physiological activities, including anticancer and anti-inflammatory effects. However, the mechanisms underlying the rejuvenation effect of Rb2 in human skin cells have not been elucidated. Methods: We performed a senescence-associated β-galactosidase staining assay to confirm cellular senescence in human dermal fibroblasts (HDFs). The regulatory effects of Rb2 on autophagy were evaluated by analyzing the expression of autophagy marker proteins, such as microtubule-associated protein 1A/1B-light chain (LC) 3 and p62, using immunoblotting. Autophagosome and autolysosome formation was monitored using transmission electron microscopy. Autophagic flux was analyzed using tandem-labeled GFP-RFP-LC3, and lysosomal function was assessed with Lysotracker. We performed RNA sequencing to identify potential target genes related to HDF rejuvenation mediated by Rb2. To verify the functions of the target genes, we silenced them using shRNAs. Results: Rb2 decreased β-galactosidase activity and altered the expression of cell cycle regulatory proteins in senescent HDFs. Rb2 markedly induced the conversion of LC3-I to LC3-II and LC3 puncta. Moreover, Rb2 increased lysosomal function and red puncta in tandem-labeled GFP-RFP-LC3, which indicate that Rb2 promoted autophagic flux. RNA sequencing data showed that the expression of DNA damage-regulated autophagy modulator 2 (DRAM2) was induced by Rb2. In autophagy signaling, Rb2 activated the AMPK-ULK1 pathway and inactivated mTOR. DRAM2 knockdown inhibited autophagy and Rb2-restored cellular senescence. Conclusion: Rb2 reverses cellular senescence by activating autophagy via the AMPK-mTOR pathway and induction of DRAM2, suggesting that Rb2 might have potential value as an antiaging agent.

• Journal of Food Hygiene and Safety
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• v.38 no.4
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• pp.202-210
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• 2023

To analyze the pesticide residues in commercial bee pollen products in South Korea, 61 samples were collected and screened for 339 pesticides. Results revealed that approximately 34% (>LOQ) of samples were contaminated with at least one pesticide. The pesticide residue detection rates of domestic and imported samples were 31% and 44%, respectively. Furthermore, the pesticide residue detection rate of online distribution (60%) was higher than that of offline distribution (27%). Fifteen pesticides were discovered in bee pollen, and pendimethalin, chlorfenvinphos, chlorpyrifos, and fluazinam were detected in 7, 6, 3, and 2 order of frequency, respectively. Even though its concentration was low, chlorfenvinphos which is banned in food crops in the United States, European Union, and Korea, was detected in bee pollen samples commonly. Therefore, continuous investigation of pesticide residues in bee pollen products and their acceptance criteria is required for safety.

• Journal of Korean Society of Transportation
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• v.23 no.2
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• pp.117-130
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• 2005

In both deteministic user Optimal Traffic Assignment Model (UOTAM) and stochastic UOTAM, travel time, which is a major ccriterion for traffic loading over transportation network, is defined by the sum of link travel time and turn delay at intersections. In this assignment method, drivers actual route perception processes and choice behaviors, which can become main explanatory factors, are not sufficiently considered: therefore may result in biased traffic loading. Even though there have been some efforts in Stochastic UOTAM for reflecting drivers' route perception cost by assuming cumulative distribution function of link travel time, it has not been fundamental fruitions, but some trials based on the unreasonable assumptions of Probit model of truncated travel time distribution function and Logit model of independency of inter-link congestion. The critical reason why deterministic UOTAM have not been able to reflect route perception cost is that the route perception cost has each different value according to each origin, destination, and path connection the origin and destination. Therefore in order to find the optimum route between OD pair, route enumeration problem that all routes connecting an OD pair must be compared is encountered, and it is the critical reason causing computational failure because uncountable number of path may be enumerated as the scale of transportation network become bigger. The purpose of this study is to propose a method to enable UOTAM to reflect route perception cost without route enumeration between an O-D pair. For this purpose, this study defines a link as a least definition of path. Thus since each link can be treated as a path, in two links searching process of the link label based optimum path algorithm, the route enumeration between OD pair can be reduced the scale of finding optimum path to all links. The computational burden of this method is no more than link label based optimum path algorithm. Each different perception cost is embedded as a quantitative value generated by comparing the sub-path from the origin to the searching link and the searched link.