• Journal of Ginseng Research
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• 제40권1호
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• pp.47-54
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• 2016

Background: The roots of Panax ginseng contain noble tetracyclic triterpenoid saponins derived from dammarenediol-II. Dammarene-type ginsenosides are classified into the protopanaxadiol (PPD) and protopanaxatriol (PPT) groups based on their triterpene aglycone structures. Two cytochrome P450 (CYP) genes (CYP716A47 and CYP716A53v2) are critical for the production of PPD and PPT aglycones, respectively. CYP716A53v2 is a protopanaxadiol 6-hydroxylase that catalyzes PPT production from PPD in P. ginseng. Methods: We constructed transgenic P. ginseng lines overexpressing or silencing (via RNA interference) the CYP716A53v2 gene and analyzed changes in their ginsenoside profiles. Result: Overexpression of CYP716A53v2 led to increased accumulation of CYP716A53v2 mRNA in all transgenic roots compared to nontransgenic roots. Conversely, silencing of CYP716A53v2 mRNA in RNAi transgenic roots resulted in reduced CYP716A53v2 transcription. HPLC analysis revealed that transgenic roots overexpressing CYP716A53v2 contained higher levels of PPT-group ginsenosides ($Rg_1$, Re, and Rf) but lower levels of PPD-group ginsenosides (Rb1, Rc, $Rb_2$, and Rd). By contrast, RNAi transgenic roots contained lower levels of PPT-group compounds and higher levels of PPD-group compounds. Conclusion: The production of PPD- and PPT-group ginsenosides can be altered by changing the expression of CYP716A53v2 in transgenic P. ginseng. The biological activities of PPD-group ginsenosides are known to differ from those of the PPT group. Thus, increasing or decreasing the levels of PPT-group ginsenosides in transgenic P. ginseng may yield new medicinal uses for transgenic P. ginseng.

• Pediatric Infection and Vaccine
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• 제8권2호
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• pp.175-180
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• 2001

목 적 : 최근 국내에서 1998년부터 5TU 대신 2TU를 사용함으로써 결과 판정에 어려움이 있다. 5TU와 2TU에 대한 투베르쿨린 반응 정도는 차이가 있게 마련이나 아직까지 적절한 국내 기준이 마련되어 있지 않은 실정이기에 생후 4주 이내에 경피 다자법으로 BCG를 접종 받은 영아에서의 PPD 1TU, 2TU 및 5TU에 대한 반응을 관찰하고자 하였다. 방 법 : 2000년 1월부터 2000년 8월까지 8개월간 순천향대학병원 육아지도회에서 결핵의 가족력, 피부 질환, 영양 장애 및 면역 기능 저하 소견이 없는 건강한 신생아 중 생후 4주 이내에 경피 다자법(Tokyo strain, Japan BCG Laboratory Co.)으로 BCG를 접종한 92명(남아 : 56명, 여아 : 36명)에게 생후 4개월에 PPD 1TU, 2TU 및 5TU로 결핵 피부반응 검사를 시행한 후 그 반응을 관찰하였다. 통계처리는 통계 프로그램인 SPSS 10.0을 이용하여 ANOVA, Chi-square test을 적용하여 P<0.05인 경우 통계적으로 유의하다고 판단하였다. 결 과 : 1) 전체 92명 중 31명에서 1TU, 31명에서 2TU 그리고 30명에서는 5TU를 사용하여 결핵 피부반응 검사를 시행하였다. 1TU로 검사시 경결의 크기가 0 mm인 경우가 가장 많았고 그 다음으로 7 mm, 6 mm 순으로 많았다. 2TU로 검사시 경우 8 mm가 가장 많았으며 그 다음은 3 mm, 6 mm, 10 mm가 많았다. 5TU로 검사시 13 mm와 6 mm가 가장 많았다. 2) 경결의 평균 크기는 PPD 1TU, 2TU 및 5TU로 검사하였을 때 각각 $5.7{\pm}4.2mm$, $7.1{\pm}3.7mm$, $9.2{\pm}4.2mm$로 1TU와 2TU는 통계학적으로 차이가 없었으나 5TU의 경우 PPD 1, 2TU에 비해 평균 경결 크기가 의미 있게 컸다(P<0.05). 3) 경결의 크기가 10 mm 이상인 경우는 1TU에서는 31명 중 6명(19.4%), 2TU에서는 31명 중 9명(29%), 5TU에서는 30명 중 16명(53.3%)이었으며 1TU와 2TU는 통계학적으로 차이가 없었으나 5TU의 경우 PPD 1, 2TU에 비해 의미 있게 많았다(P<0.05). 경결의 크기가 5 mm 이상인 경우는 통계학적 유의성은 없었지만 5TU로 검사시 가장 많았다. BCG 반흔 수는 1TU의 경우 $13.81{\pm}4.48$개, 2TU는 $13.06{\pm}4.18$개, 5TU는 $13.07{\pm}3.07$개로 차이가 없었다. 결 론 : 현재 국내에서 사용되고 있는 2TU을 사용한 결핵 피부반응 검사에 대한 새로운 평가 기준이 필요하고 결핵의 진단기준에 대한 더 많은 연구가 필요하다고 사료된다.

• 대한수의학회지
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• 제16권2호
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• pp.151-158
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• 1976

The tuberculin test have been widely utilized to diagnose tuberculosis of the dairy cattle. It was reported that approximately 70% of the dairy cattle with positive tuberculin reaction in Korea had been non-pathogenic lesion. So the studies on the specific method to diagnose tuberculosis of them is really need in Korea. Therefore this study investigated upon the diagnostic method for cattle tuberculosis in the aspect of cellular-immunology. The results obtained in this investigation are summarized as follows: 1. All the tuberculin tests to the swine inoculated with BCG.(B group), Mycobacterium avium (A group) and Mycobacterium smegmatis (S group), respectively, were represented high positive reaction and were no differences in the species of inoculated bacteria. 2. In the migration inhibition test using Iymphocyte in circulating blood of the swine inoculated with three species of acid-fast bacteria respectively, the test to A group was represented slight positive for migration in the presence of $15{\mu}g/ml$-PPD and the other reaction were clear and total positive for migration inhibition in the presence of $25{\mu}g/ml$-PPD or more, and the test to B group was the same results as to A group approximately. The test to S group was represented slight positive for migration inhibition in the presence of $15{\mu}g/ml$-PPD, but the results were the same in the presence of $25{\mu}g/ml$-PPD and $35{\mu}g/ml$-PPD. These results showed that there were remarkable differences between group A, B and group S in the presence of $25{\mu}g/ml$-PPD and $35{\mu}g/ml$-PPD. 3. The transformation rates of lymphocyte in PPD treated tissue culture had no significance without any relation between PPD treatment and non-treatment but A group and B group showed significance. And A group and B group showed high significance in comparison with N group and S group in the LSD examination. 4. The infection rated in lymphocyte of BCG inoculated after 3 days tissue culturing were represented those of high infection but its cellular degeneration rates almost did not change. The infection rates in bacilli in N group and S group were low but after 3 days inoculation it shewed higher cellular degeneration.

• 사회복지연구
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• 제40권3호
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• pp.299-326
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• 2009

본 연구는 정신장애인이 지각하는 사회의 편견에 영향을 미치는 요인은 무엇이며 이러한 요인의 영향은 어떠한 이론적 근거에서 설명될 수 있는지 규명하는데 목적이 있다. 상징적 상호작용주의이론에 근거하여 직업적 역할, 직업재활서비스욕구, 가족의 편견인식이 정신장애인이 지각하는 편견에 미치는 영향을 검정하였다. 본 연구의 분석에 활용된 자료는 2008년 국가인권위원회의 '전국 정신질환자 및 가족 생활실태조사'자료 중 충실히 응답된 229사례였고, 자료분석은 위계적 회귀분석을 활용하였다. 자료분석결과 정신장애인의 증상이 높을수록 편견인식이 높았고, 직업적 역할이 있으면 편견인식이 유의하게 감소하였다. 직업을 통해 재활을 도모하는 상태에 있으면 편견인식이 유의하게 증가하였고, 가족의 편견인식이 높으면 정신장애인의 편견인식도 높은 것으로 나타났다. 또한 가족의 편견인식은 직업적 역할이 정신장애인의 편견인식을 감소시키는 효과를 상쇄하는 것으로 나타났다. 이러한 연구결과에 근거하여 이론적, 실천적 함의를 논의하였다.

• Journal of Ginseng Research
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• 제45권2호
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• pp.325-333
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• 2021

Background: Alzheimer's disease (AD) is one of the most prevalent neurodegenerative disorders. Enhancing hippocampal neurogenesis by promoting proliferation and differentiation of neural stem cells (NSCs) is a promising therapeutic strategy for AD. 20(S)-protopanaxadiol (PPD) and oleanolic acid (OA) are small, bioactive compounds found in ginseng that can promote NSC proliferation and neural differentiation in vitro. However, it is currently unknown whether PPD or OA can attenuate cognitive deficits by enhancing hippocampal neurogenesis in vivo in a transgenic APP/PS1 AD mouse model. Here, we administered PPD or OA to APP/PS1 mice and monitored the effects on cognition and hippocampal neurogenesis. Methods: We used the Morris water maze, Y maze, and open field tests to compare the cognitive capacities of treated and untreated APP/PS1 mice. We investigated hippocampal neurogenesis using Nissl staining and BrdU/NeuN double labeling. NSC proliferation was quantified by Sox2 labeling of the hippocampal dentate gyrus. We used western blotting to determine the effects of PPD and OA on Wnt/GSK3β/β-catenin pathway activation in the hippocampus. Results: Both PPD and OA significantly ameliorated the cognitive impairments observed in untreated APP/PS1 mice. Furthermore, PPD and OA significantly promoted hippocampal neurogenesis and NSC proliferation. At the mechanistic level, PPD and OA treatments resulted in Wnt/GSK-3β/β-catenin pathway activation in the hippocampus. Conclusion: PPD and OA ameliorate cognitive deficits in APP/PS1 mice by enhancing hippocampal neurogenesis, achieved by stimulating the Wnt/GSK-3β/β-catenin pathway. As such, PPD and OA are promising novel therapeutic agents for the treatment of AD and other neurodegenerative diseases.

• Journal of Ginseng Research
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• 제47권4호
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• pp.572-582
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• 2023

Background: Free fatty acid-induced lipotoxicity is considered to play an important role in pancreatic β-cell dysfunction. The effect of ginsenosides on palmitic acid-induced pancreatic beta-cells cell death and failure of glucose-stimulated secretion of insulin (GSIS) was evaluated in this study. Methods: Enzyme-linked immunosorbent assay kit for a rat insulin was used to quantify glucose-stimulated insulin secretion. Protein expression was examined by western blotting analysis. Nuclear condensation was measured by staining with Hoechst 33342 stain. Apoptotic cell death was assessed by staining with Annexin V. Oil Red O staining was used to measure lipid accumulation. Results: We screened ginsenosides to prevent palmitic acid-induced cell death and impairment of GSIS in INS-1 pancreatic β-cells and identified protopanaxadiol (PPD) as a potential therapeutic agent. The protection effect of PPD was likely due to a reduction in apoptosis and lipid accumulation. PPD attenuated the palmitic acid-induced increase in the levels of B-cell lymphoma-2-associated X/B-cell lymphoma 2, poly (ADP-ribose) polymerase and cleaved caspase-3. Moreover, PPD prevented palmitic acid-induced impairment of insulin secretion, which was accompanied by an increase in the activation of phosphatidylinositol 3-kinase, peroxisome proliferator-activated receptor γ, insulin receptor substrate-2, serine-threonine kinase, and pancreatic and duodenal homeobox-1. Conclusion: Our results suggest that the protective effect of PPD on lipotoxicity and lipid accumulation induced by palmitic acid in pancreatic β-cells.

• 대한인간공학회지
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• 제17권3호
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• pp.41-59
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• 1998

Thermal comfort aspect of a locally-cooled target space in warm and humid environments(typically in the rainy summer season) was studied in view of PPD index. First. theoretical analyses were conducted to examine the effect of the governing parameters(such as air temperature, relative humidity and air velocity, etc.) using a computer model. Secondly, experimental investigations were also performed in a climatic room designed to simulate corresponding thermal conditions of outdoor environments. During the tests, temporal variation of PPD was recorded as functions of climatic variables(outdoor and indoor temperatures, relative humidity and air velocity) for the given human factors(metabolic heat generation and clothing). From both theoretical and experimental investigations, air temperature and air velocity were found to be the most dominant parameters affecting PPD of the target space. Results were summarized as: 1. Relative humidity of the locally-cooled target space tends to approach that of outdoor's as the space is subjected to an ON-OFF mode of cooling, since moisture potential of the two rooms reaches an equalized state as a result of moisture diffusion. 2. It was recognized that changes in relative humidity did not show any significance in view of thermal comfort as was reported in the previous studies, while variations of both temperature and air velocity caused relatively large changes in the degree of thermal comfort. 3. In-door environment should be evaluated in terms of PPD instead of relative humidity commonly recognized as an important climatic variable particularly in warm and humid environments.

• Bulletin of the Korean Chemical Society
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• 제25권8호
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• pp.1195-1201
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• 2004

Tyrosinase was covalently immobilized on platinum electrode according to the method we developed for laccase (Bull. Korean Chem. Soc. 2002, 23(7), 385) and p-chlorophenol, p-cresol, and phenol could be detected with sensitivities of 334, 139 and 122 nA/ ${\mu}M$ and the detection limits of 1.0, 2.0, and 2.5 ${\mu}M$, respectively. The response time ($t_{90\%}$) is 3 seconds for p-chlorophenol, and 5 seconds for p-cresol and phenol. The optimal pHs of the sensor are in the range of 5.0- 6.0. This sensor can tolerate at least 500 times repeated injections of p-chlorophenol with retaining 80% of initial activity. In case of tyrosinase and laccase co immobilized platinum electrode, the sensitivities are 560 nA/ ${\mu}M$ for p-phenylenediamine (PPD) and 195 nA/ ${\mu}M$ for p-chlorophenol, respectively. The sensitivity of the bi-enzyme sensor for PPD increases 70% compared to that of only laccase immobilized one, but the sensitivity for p-chlorophenol decreases 40% compared to that of only tyrosinase immobilized one. The sensitivity increase for the bi-enzyme sensor for PPD can be ascribed to the additional catalytic function of the co-immobilized tyrosinase. The sensitivity decrease for p-chlorophenol can be explained by the “blocking effect” of the co-immobilized laccase, which hinders the mass transport through the immobilized layer. If PPD was detected with the electrode that had been used for p-chlorophenol, the sensitivity decreased 20% compared to that of the electrode that had been used only for PPD. Similarly, if p-chlorophenol was detected with PPD detected electrode, the sensitivity also decreased 20%. The substrate-induced conformation changes of the enzymes in a confined layer may be responsible for the phenomena.

• Journal of Ginseng Research
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• 제46권4호
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• pp.505-514
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• 2022

Background: The roots of Panax ginseng contain two types of tetracyclic triterpenoid saponins, namely, protopanaxadiol (PPD)-type saponins and protopanaxatiol (PPT)-type saponins. In P. ginseng, the protopanaxadiol 6-hydroxylase (PPT synthase) enzyme catalyses protopanaxatriol (PPT) production from protopanaxadiol (PPD). In this study, we constructed homozygous mutant lines of ginseng by CRISPR/Cas9-mediated mutagenesis of the PPT synthase gene and obtained the mutant ginseng root lines having complete depletion of the PPT-type ginsenosides. Methods: Two sgRNAs (single guide RNAs) were designed for target mutations in the exon sequences of the two PPT synthase genes (both PPTa and PPTg sequences) with the CRISPR/Cas9 system. Transgenic ginseng roots were generated through Agrobacterium-mediated transformation. The mutant lines were screened by ginsenoside analysis and DNA sequencing. Result: Ginsenoside analysis revealed the complete depletion of PPT-type ginsenosides in three putative mutant lines (Cr4, Cr7, and Cr14). The reduction of PPT-type ginsenosides in mutant lines led to increased accumulation of PPD-type ginsenosides. The gene editing in the selected mutant lines was confirmed by targeted deep sequencing. Conclusion: We have established the genome editing protocol by CRISPR/Cas9 system in P. ginseng and demonstrated the mutated roots producing only PPD-type ginsenosides by depleting PPT-type ginsenosides. Because the pharmacological activity of PPD-group ginsenosides is significantly different from that of PPT-group ginsenosides, the new type of ginseng mutant producing only PPD-group ginsenosides may have new pharmacological characteristics compared to wild-type ginseng. This is the first report to generate target-induced mutations for the modification of saponin biosynthesis in Panax species using CRISPR-Cas9 system.

• Journal of Ginseng Research
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• 제47권4호
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• pp.543-551
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• 2023

Background: Panax ginseng Meyer is a representative Chinese herbal medicine with antioxidant and anti-inflammatory activity. 20(S)-Protopanaxadiol (PPD) has been isolated from ginseng and shown to have promising pharmacological activities. However, effects of PDD on pulmonary fibrosis (PF) have not been reported. We hypothesize that PDD may reverse inflammation-induced PF and be a novel therapeutic strategy. Methods: Adult male C57BL/6 mice were used to establish a model of PF induced by bleomycin (BLM). The pulmonary index was measured, and histological and immunohistochemical examinations were made. Cell cultures of mouse alveolar epithelial cells were analyzed with Western blotting, coimmunoprecipitation, immunofluorescence, immunohistochemistry, siRNA transfection, cellular thermal shift assay and qRT-PCR. Results: The survival rate of PPD-treated mice was higher than that of untreated BLM-challenged mice. Expression of fibrotic hallmarks, including α-SMA, TGF-β1 and collagen I, was reduced by PPD treatment, indicating attenuation of PF. Mice exposed to BLM had higher STING levels in lung tissue, and this was reduced by phosphorylated AMPK after activation by PPD. The role of phosphorylated AMPK in suppressing STING was confirmed in TGF-b1-incubated cells. Both in vivo and in vitro analyses indicated that PPD treatment attenuated BLM-induced PF by modulating the AMPK/STING signaling pathway. Conclusion: PPD ameliorated BLM-induced PF by multi-target regulation. The current study may help develop new therapeutic strategies for preventing PF.