청가시덩굴 추출물의 기능성 원료 표준화를 위한 지표성분 Resveratrol, trans-Scirpusin A의 분석법 개발 및 검증

권진관(Jin Gwan Kwon),정연우(Yeon Woo Jung),최윤혁(Yun-Hyeok Choi),이지은(Ji Eun Lee),정원식(Wonsik Jeong),이정아(Jung A Lee),최춘환(Chun Whan Choi),안은경(Eun-Kyung Ahn),최용문(Yongmun Choi),홍성수(Seong Su Hong) 한국식품영양과학회 2022 한국식품영양과학회지 Vol.51 No.11

본 연구는 HPLC를 이용하여 청가시덩굴 추출물을 개별인정형 건강기능식품의 기능성 원료로 개발하기 위한 원료 표준화의 일환으로, 청가시덩굴 추출물의 지표성분을 resveratrol과 trans-scirpusin A로 설정하고 이에 대한 HPLC 분석법을 확립하여 유효성의 검증을 실시하였다. 분석법 유효성 검증은 특이성, 직선성, 정확도, 정밀도, 검출한계 및 정량한계 등을 통해 분석법의 신뢰성을 검증하였으며, 그 결과 표준용액과 청가시덩굴 추출물 간의 HPLC 크로마토그램 및 UV spectrum의 일치 여부 등의 비교를 통해 다른 물질과 간섭 없이 피크가 분리된 것으로 특이성을 확인하였다. 또한 표준용액 검량선의 상관계수(R²)는 0.9999로 매우 우수한 직선성으로 관찰되어 분석에 적합한 것으로 확인되었으며, 검량선의 기울기 및 표준편차를 이용한 검출한계는 resveratrol이 0.98 μg/mL, trans-scirpusin A는 0.49 μg/mL였고 정량한계는 resveratrol이 2.98 μg/mL, trans-scirpusin A는 1.48 μg/mL로 각각 확인되었다. 청가시덩굴 추출물에 표준물질을 3개 농도 첨가하고 분석한 회수율은 resveratrol이 98.77~99.24%, trans-scirpusin A는 98.45~99.45%로 나타나 정확성이 있는 것을 확인할 수 있었다. 청가시덩굴 추출물의 조제 농도 2.2, 4.4 및 6.6 mg/mL에서 반복성은 resveratrol이 0.99~1.22%, trans-scirpusin A는 1.12~1.32%를, 실험실 내 정밀성에서는 일내 정밀성은 resveratrol이 0.67~0.87%, trans-scirpusin A는 1.18~1.33%로 나타났고 일간 정밀성은 resveratrol이 0.93~1.22%, trans-scirpusin A는 1.33~2.27%로 확인되어 본 분석법은 정밀성이 있음을 확인할 수 있었다. 이상의 분석결과를 통해 확립된 청가시덩굴 추출물의 지표성분인 resveratrol과 trans-scirpusin A의 HPLC 분석법은 적합한 시험법으로 검증되었으며, 본 시험법은 향후 청가시덩굴 추출물의 건강기능식품 기능성 원료 개발과 표준화를 위한 기초자료로 활용될 것으로 사료된다. This study was undertaken to establish an analytical method using high-performance liquid chromatography (HPLC). HPLC for the standard determination of resveratrol and trans-scirpusin A as functional ingredients in Smilax sieboldii extract. We evaluated the specificity, linearity, accuracy, precision, limit of detection (LOD), and limit of quantitation (LOQ) of various analytical methods for detecting resveratrol and trans-scirpusin A using HPLC. The specificity was confirmed by the chromatogram obtained using the HPLC analytical method. Also, the results of UV and the coefficient of correlation (R²) obtained was 0.999, which confirmed that this was a suitable analysis with high linearity. The LOD was 0.98, 0.49 μg/mL, and LOQ was 2.98, 1.48 μg/mL, which was confirmed as a suitable limit level for the analysis of resveratrol and trans-scirpusin A content in the S. sieboldii extract. The recovery of resveratrol and trans-scirpusin A content was determined to be 98.77±0.73∼99.24±1.47% and 98.45±1.18∼99.45±1.66%, respectively, indicating high accuracy. The intra-day repeatability and the intra-laboratory precision of the daily repetition were confirmed to be 0.67∼0.87%, 1.18∼1.33% and 0.93∼1.22%, 1.33∼2.27%, respectively, for trans-scirpusin A, for the relative standard deviation. These results indicate that the reported HPLC method is simple, reliable, and reproducible for the detection of resveratrol and trans-scirpusin A in S. sieboldii extract.

Phorbaketal A stimulates osteoblast differentiation through TAZ mediated Runx2 activation

Byun, Mi Ran,Kim, A Rum,Hwang, Jun-Ha,Sung, Mi Kyung,Lee, Yeon Kyung,Hwang, Buyng Su,Rho, Jung-Rae,Hwang, Eun Sook,Hong, Jeong-Ho Elsevier 2012 FEBS letters Vol.586 No.8

<P><B>Highlights</B></P><P>► Phorbaketal A stimulates osteogenesis. ► Phorbaketal A increases TAZ-mediated osteoblast marker gene expression. ► ERK is an important kinase for phorbaketal A induced osteogenic differentiation. ► Phorbaketal A is suggested as a potential compound for stimulating bone formation.</P> <P><B>Abstract</B></P><P>Osteoporosis arises from an imbalance between osteoblastic bone formation and osteoclastic bone resorption. In this study, we screened molecules from marine natural products that stimulate osteoblast differentiation. We found that phorbaketal A significantly stimulates osteoblast differentiation in mesenchymal cells. Increased interaction of TAZ and Runx2 stimulated phorbaketal A-induced expression of osteoblastic marker genes. The activation of ERK was important for the stimulation of differentiation because an inhibitor of ERK blocked phorbaketal A-induced osteogenic differentiation. Taken together, the results showed that phorbaketal A stimulates TAZ-mediated osteoblast differentiation through the activation of ERK.</P><P><B>Structured summary of protein interactions</B></P><P><B>TAZ</B> physically interacts with <B>RUNX2</B> by pull down (View interaction)</P>

3T3-L1 지방세포에서 초과 에탄올 추출물의 항비만 효과

이정아 ( Jung A Lee ),박영진 ( Young Jin Park ),정원식 ( Wonsik Jeong ),홍성수 ( Seong Su Hong ),안은경 ( Eun-kyung Ahn ) 한국응용생명화학회 2017 Journal of Applied Biological Chemistry (J. Appl. Vol.60 No.1

Amomum tsao-ko used as a traditional oriental herbal medicine, is indigenous to several Asia countries. In this study, we investigated anti-obesity activity of the ethanol extract of Amomum Taso-ko (A. tsao-ko). The ethanol extract of A. tsao-ko inhibited adipocyte differentiation using Oil Red O assay in 3T3-L1 cells. Inhibitory effect of the ethanol extract of A. tsao-ko on adipogenesis was modulated by down-regulation adipogenic transcriptional factor such as peroxisome proliferator-activated receptor γ (PPARγ), CCAAT-enhancer-binding protein α (C/EBPα) and suppressed expression of fatty acid synthase, aP2, and resistin. We demonstrated that A. tsao-ko significantly inhibited adipogenesis and reduced PPARγ and C/EBPα expression in a dose-dependent manner. These results suggest that A. tsao-ko has an anti-obesity effect by inhibition of adipogenic transcription factor and adipocyte-specific genes in 3T3-L1 cells.

Pheophorbide a-mediated photodynamic therapy induces autophagy and apoptosis via the activation of MAPKs in human skin cancer cells

YOON, HYO-EUN,OH, SEONE-HEE,KIM, SOO-A,YOON, JUNG-HOON,AHN, SANG-GUN Spandidos Publications 2014 Oncology reports Vol.31 No.1

Pheophorbide a (Pa), a chlorophyll derivative, is a photosensitizer that can induce significant antitumor effects in several types of tumor cells. The present study investigated the mechanism of Pa-mediated photodynamic therapy (Pa-PDT) in the human skin cancer cell lines A431 and G361. PDT significantly inhibited the cell growth in a Pa-concentration-dependent manner. We observed increased expression of Beclin-1, LC3B and ATG5, which are markers of autophagy, after PDT treatment in A431 cells but not in G361 cells. In G361 cells, Pa-PDT strongly induced PARP cleavage and subsequent apoptosis, which was confirmed using Annexin V/Propidium iodide double staining. Pa-PDT predominantly exhibited its antitumor effects via activation of ERK1/2 and p38 in A431 and G361 cells, respectively. An in vivo study using the CAM xenograft model demonstrated that Pa-PDT strongly induced autophagy and apoptosis in A431-transplanted tumors and/or apoptosis in G361-transplanted tumors. These results may provide a basis for understanding the underlying mechanisms of Pa-PDT and for developing Pa-PDT as a therapy for skin cancer.

Evaluating the TaqMan Jr a -Genotyping Method for Rapidly Predicting the Presence of Anti-Jr a Antibodies

Koo Yu-Kyung,Kwon Soon Sung,Suh Eun Jung,Kim Na Hyeong,Kim Hyun Kyung,Cho Youn Keong,Choi Seung Jun,Kim Sinyoung,Lee Kyung-A 대한진단검사의학회 2024 Annals of Laboratory Medicine Vol.44 No.5

Background: The Jra antigen is a high-prevalence red blood cell (RBC) antigen. Reports on cases of fatal hemolytic disease of the fetus and newborn and acute hemolytic transfusion reactions suggest that antibodies against Jra (anti-Jra) have potential clinical significance. Identifying anti-Jra is challenging owing to a lack of commercially available antisera. We developed an alternative approach to rapidly predict the presence of anti-Jra using the Taq- Man single-nucleotide polymorphism (SNP)-genotyping method. Methods: Residual peripheral blood samples from 10 patients suspected of having the anti-Jra were collected. Two samples with confirmed Jr(a–) RBCs and anti-Jra were used to validate the TaqMan genotyping assay by comparing the genotyping results with direct sequencing. The accuracy of the assay in predicting the presence of anti-Jra was verified through crossmatching with in-house Jr(a–) O+ RBCs. Results: The TaqMan-genotyping method was validated with two Jr(a–) RBC- and anti-Jraconfirmed samples that showed concordant Jra genotyping and direct sequencing results. Jra genotyping for the remaining samples and crossmatching the serum samples with inhouse Jr(a–) O+ RBCs showed consistent results. Conclusions: We validated a rapid, simple, accurate, and cost-effective method for predicting the presence of anti-Jra using a TaqMan-based SNP-genotyping assay. Implementing this method in routine practice in clinical laboratories will assist in solving difficult problems regarding alloantibodies to high-prevalence RBC antigens and ultimately aid in providing safe and timely transfusions and proper patient care.

한국인에서 혈소판 당단백 Ⅱb/Ⅲa 유전자 다형성과 관동맥 성형술 후 재 협착과의 관계

이민수,이정우,김보영,임대승,강정아,김정희,김윤철,성보영,최성준,성인환,전은석 충남대학교 의과대학 지역사회의학연구소 2000 충남의대잡지 Vol.27 No.2

Platelet aggregation is the final pathway of acute coronary syndrome such as acute myocardial infarction and unstable angina. Platelet glycoprotein IIb/IIIa is a membrane receptor for fibrinogen and yon Willebrand factor and it plays an important role in platelet aggregation and in the pathogenesis of acute coronary syndrome. It is known that polymorphism of the gene that encoding platelet glycoprotein IIb/IIIa(PI^A1/A2) is strongly related to acute coronary syndrome in Caucasian, but not in Koreans. We investigated relationship between platelet glycoprotein llb/Illa gene polymorphism and restenosis of coronary artery after angioplasty in Koreans. Total 371 patients(M=251. F=120) were enrolled. Angioplasty group comprised 143 patients who underwent coronary angioplasty, and in the angioplasty group, restenosis group comprised with the 65 patients who had restenotic lesion over 50% of luminal diameter in follow-up coronary angiography. Normal group comprised 153 patients who had no significant angiographic lesion and variant angina group comprised 75 patients who were positive in ergonovine test. Genomic DNA was extracted from peripheral arterial blood. To determine the frequency of P1^A1/A2 genotype, polymerase chain reaction(PCR) was done and the product was restricted with Mspl. 3%. agarrose gel electrophoresis showed restriction fragment length polymorphism. Clinical profile and risk factor were also reviewed. Among all 371 patients of study group, genotype of only one patients in restenosis group if is proven to be PI^A1/A2 heterozygote. All patients of normal study group, no restenosis group, and the other patients in restenosis group have an PI^A1 homozygote genotype. In our study, platelet glycoprotein IIb/Illa polymorphism has no relationship with restenosis of the coronary artery after angioplasty in Koreans. But the genotypic frequency of platelet glycoprotein IIb/IIIa gene polymorphism in Koreans is concordant with that of previous studies.

CMOS Transistors with a 70-nm Gate Length for 0.13-μm-Node High-Performance Applications

Kyoung-Seok Rha,Dong-Hun Lee,Eun-Seung Jung,Hae-Kyung Kong,Hyae-Ryoung Lee,Jeong-Ho Lyu,Jeong-Hwan Yang,Jin-Suk Jung,Jung-A Choi,Kwang-Pyuk Suh,Young-Wug Kim 한국물리학회 2004 THE JOURNAL OF THE KOREAN PHYSICAL SOCIETY Vol.44 No.1

Conventional CMOS transistors for high performance with a 70-nm physical gate length were fabricated to evaluate the 130-nm-technology node. In this work, we enhanced the performance of transistors by using high-dose P+ implantation, a plasma-nitridation gas oxide, a hydrogen prebake (HPB), and a mechanical stress layer. We also reduced the overlap capacitances by using an oset spacer. The NMOS and PMOS have drive currents equal to 880 A/m and 420 A/m, respectively, with Ioff = 10 nA/m at Vdd = 1.2 V. The unit delay of the ring oscillator (1 fan-out) was 11.5 ps.

한국인에서 5,10-Methylenetetrahydrofolate Reductase 유전자 다형성 분석 및 임신의 결과에 미치는 영향

김아리 ( Ari Kim ),강은지 ( Eun Ji Kang ),이경아 ( Kyung A Lee ),박세나 ( Se Na Park ),박종순 ( Jong Soon Park ),박보현 ( Bo Hyun Park ),박혜숙 ( Hyesook Park ),박미혜 ( Mi Hye Park ),전선희 ( Sun Hee Chun ),안정자 ( Jung Ja Ahn ) 대한주산의학회 2006 Perinatology Vol.17 No.3

Objective: The purpose of this study was to analyze MTHFR polymorphism among the Korean population and to evaluate the relationship between serum levels of homocysteine and MTHFR polymorphism and also to investigate the effect on pregnancy outcomes. Methods: DNA was extracted from whole blood of 600 pregnant women. All samples were genotyped for the C677T and A1298C polymorphisms in MTHFR gene by PCR-RELP assay. Serum levels of homocysteine and folate were measured by high performance liquid chromatography for homocysteine and radioassay for folate. Pregnancy outcomes were estimated by gestational weeks and birth weights of newborns. Results: Serum homocysteine was higher in women with the T/T genotype than those with the C/T or C/C genotype of the MTHFR C677T polymorphism (p<0.05). And also serum homocysteine was higher in women with the A/A genotype than those with the A/C or C/C genotype of the MTHFR A1298C polymorphism (p<0.05). Serum homocysteine was negatively correlated with serum folate in all MTHFR genotypes, especially prominent in T/T genotype of MTHFR C677T polymorphism and A/A genotype of MTHFR A1298C polymorphism. Gestational age and the birth weight of infant from hyperhomocysteinemic mothers whose homocysteine levels higher than 15 μmol/L were 36.1 weeks, 3053.8 g, respectively, which were significant lower than those from normohomocysteinemic mothers (38.3 weeks, 3,215.3 g) (p<0.05). Conclusion: Serum homocysteine was influenced significantly by MTHFR C677T polymorphism and MTHFR A1298C polymorphism. MTHFR C677T and A1298C polymorphism and serum homocysteine levels affect pregnancy outcomes, although not mainly by serum folate level.

모 제약회사 근로자들의 직업성 천식 유병률과 약제 분말 노출수준

오성수,최용휴,김은아,이영목,장재길,유장진,박승현,김태균,정수영,김규상,강성규 大韓産業醫學會 2006 대한직업환경의학회지 Vol.18 No.2

목적: 제약회사에서 직업상 천식의 유병 규모를 파악하고 작업공정에 대한 작업환경 측정을 통해 제약회사의 약제 분말에 대한 노출수준을 평가하고자 하였다. 방법: 제약회사에서 약제 분말에 노출되는 근로자를 대상으로 천식을 선별하기 위하여 설문조사 및 진찰, 혈액검사, 폐기능 검사 그리고 피부단자시험을 실시하였고, 각 부서별 공정에 따라서 총분진과 항생제 또는 효소제 분말에 대한 작업환경 측정을 실시하였다. 직업상 천식이 의심되는 근로자에 대하여 기관지과민성에 대한 검사(메타콜린을 이용한 비특이 기관지유발시험)와 약제 분말 기관지유발시험을 실시하여 확진하였다. 결과: 설문조사 및 진찰에서 작업 관련 천식 증상이 있는 근로자는 11명(34.4%)이었고, 약제 분말을 이용한 피부단자시험에서 1+ 이상 반응을 보인 근로자 10명(31.2%)이었다. 비특이 기관지유발시험 결과 8명(25.0%)이 양성(PC20 ≤16 mg/ml)이었다. 약제 분말 기관지유발시험을 통해 직업상 천식으로 확진된 근로자는 5명(15.6%)이었다. 작업환경 측정에서 원료를 충진하거나 투입하는 작업 그리고 정립작업에서 약제 분말에 대한 노출이 많았다. 결론: 제약회사에서 직업상 천식은 주로 제조공정 중 약제 분말에 노출된 근로자에서 발생하여, 증상 및 직업성 천식의 유병률이 높았다. 따라서 원료를 충진 또는 투입하는 작업과 정립작업 등 제조과적에서 약제 분말 노출을 줄이기 위한 세심한 주의가 필요하여, 약제 분말을 취급하는 근로자에 대해서는 천식 증상 조사뿐 아니라 정기적으로 피부단자시험, 비특이 기관지유발시험 등을 실시하여야 할 것이다. Objectives: After the investigation of one worker with occupational asthma, we surveyed the preva- lence of occupational asthma and the exposure level of pharmaceutical dust of 32 workers in a pharmaceutical company. Methods: Thirty-two of the 90 employees participated in the survey which consisted of questionnaire, blood sampling, spirometry and skin prick tests with 8 common allergens as well as 9 antibiotics and 2 enzymes. Various indices of the working environment were also measured. Subjects who had a symptom suggestive of work-related asthma or positive skin prick test were further investigated by PC20 methacholine. Nine subjects who had a PC20 result of 16 mg/ml or less (n=7) or had work-related symptoms and positive skin prick test (n=2) were referred to undergo a specific bronchial provocation test to phar- maceutical dust in an academic allergic disease center. Results: Eleven of 32 workers (34.4%) had a work-related symptom suggestive of occupational asthma. Ten (31.2%) showed positive skin prick test. 8 (25.0%) had a PC20 result of 16 mg/ml or less (indicative of significant bronchial hyperresponsiveness), and 5 (15.6%) had a positive result on the specific bronchial provocation test. Exposure levels of stuffing, input of raw materials and screening process were relatively high. Conclusions: This survey showed that pharmaceutical workers have an increased risk of occupational asthma. Although pharmaceutical factories maintain a relatively good working environment, careful control of respiratory tract exposures, especially during stuffing, input or raw materials and screening process, is important to prevent occupational asthma. Pharmaceutical workers need to undergo regular skin prick and methacholine bronchial provocation tests, as well as asthmatic symptom survey, to ensure the early detection and prevention of occupational asthma.

Intestinal Nematodes from Small Mammals Captured near the Demilitarized Zone, Gyeonggi Province, Republic of Korea

Deok-Gyu Kim,Jae-Hwan Park,Jae-Lip Kim,Bong-Kwang Jung,Sarah Jiyoun Jeon,Hyemi Lim,Mi Youn Lee,Eun-Hee Shin,Terry A. Klein,Heung-Chul Kim,Sung-Tae Chong,Jin-Won Song,Luck-Ju Baek,Jong-Yil Chai 대한기생충학열대의학회 2015 Parasites, Hosts and Diseases Vol.53 No.1

A total of 1,708 small mammals (1,617 rodents and 91 soricomorphs), including Apodemus agrarius (n = 1,400), Microtus fortis (167), Crocidura lasiura (91), Mus musculus (32), Myodes (= Eothenomys) regulus (9), Micromys minutus (6), and Tscherskia (= Cricetulus) triton (3), were live-trapped at US/Republic of Korea (ROK) military training sites near the demilitarized zone (DMZ) of Paju, Pocheon, and Yeoncheon, Gyeonggi Province from December 2004 to December 2009. Small mammals were examined for their intestinal nematodes by necropsy. A total of 1,617 rodents (100%) and 91 (100%) soricomorphs were infected with at least 1 nematode species, including Nippostrongylus brasiliensis, Heligmosomoides polygyrus, Syphacia obvelata, Heterakis spumosa, Protospirura muris, Capillaria spp., Trichuris muris, Rictularia affinis, and an unidentified species. N. brasiliensis was the most common species infecting small mammals (1,060; 62.1%) followed by H. polygyrus (617; 36.1%), S. obvelata (370; 21.7%), H. spumosa (314; 18.4%), P. muris (123; 7.2%), and Capillaria spp. (59; 3.5%). Low infection rates (0.1-0.8%) were observed for T. muris, R. affinis, and an unidentified species. The number of recovered worms was highest for N. brasiliensis (21,623 worms; mean 20.4 worms/infected specimen) followed by S. obvelata (9,235; 25.0 worms), H. polygyrus (4,122; 6.7 worms), and H. spumosa (1,160; 3.7 worms). A. agrarius demonstrated the highest prevalence for N. brasiliensis (70.9%), followed by M. minutus (50.0%), T. triton (33.3%), M. fortis (28.1%), M. musculus (15.6%), C. lasiura (13.2%), and M. regulus (0%). This is the first report of nematode infections in small mammals captured near the DMZ in ROK.