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Table of Contents
CONFERENCE ABSTRACTS
Year : 2019  |  Volume : 5  |  Issue : 4  |  Page : 277-296

7th Annual meeting of the good practice in traditional chinese medicine research association


Date of Web Publication3-Dec-2019

Correspondence Address:
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/2311-8571.271970

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How to cite this article:
. 7th Annual meeting of the good practice in traditional chinese medicine research association. World J Tradit Chin Med 2019;5:277-96

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. 7th Annual meeting of the good practice in traditional chinese medicine research association. World J Tradit Chin Med [serial online] 2019 [cited 2019 Dec 8];5:277-96. Available from: http://www.wjtcm.net/text.asp?2019/5/4/277/271970




  Astragaloside IV Protects against Autophagy and Apoptotic Death of H9c2 Cells in Myocardial Ischemia/Reperfusion Injury Top


Jing-Jing Yanga, Xu-Hui Zhangb, Wen-Jun Duanc, Xiao-Hui Mac, Neng-Gui Xud, Yong-Jun Chend, Lei Liangc

aSchool of Traditional Chinese Medicine, Guangdong Pharmaceutical University,bDepartment of Oncology, Guangdong Second Provincial General Hospital,cCollege of Pharmacy, Jinan University,dSouth China Research Center for Acupuncture and Moxibustion, Medical College of Acu-Moxi and Rehabilitation, Guangzhou University of Chinese Medicine, Guangzhou, People's Republic of China

Correspondence:

Lei Liang, E-mail: leiliang@jnu.edu.cn

Yongjun Chen, E-mail: ychen@gzucm.edu.cn

Introduction (Includes the Aim of the Study): Myocardial ischemia/reperfusion injury (MI/RI) primarily results from recovered blood flow following ischemia in the myocardial tissue, resulting in cardiac insufficiency, cell death, and arrhythmia. Astragaloside IV (Ast-IV) was reported to have effects on improving cardiac function after ischemia, but the mechanisms are poorly understood. The transcription factor GATA-4 is indispensable for physiological function of heart.

Materials and Methods: Here, we investigated the ability of Ast-IV to inhibit apoptosis and autophagy via GATA-4 as a potential mechanism underlying its protection against doxorubicin-induced cardiomyocytes death.

Results: Hypoxia/reoxygenation (H/R) markedly increased the apoptosis rate and the number of autophagosomes formed in cells, whereas Ast-IV reversed these changes. Mechanistically, Ast-IV up-regulated gene expression of the survival factor Bcl-2, p62 and suppressed H/R-induced activation of apoptosis- and autophagy-related genes (PARP, Caspase-3, Beclin-1, and LC3-II), which may likely be responsible for up-regulating expression of GATA-4 that enhanced the interactions between Bcl-2 and Beclin-1. Silencing GATA-4 expression blocked the effect of Ast-IV, whereas overexpressing this transcription factor inhibited these processes.

Conclusions: Together, these findings suggest that Ast-IV reduced MI/RI in rat cardiomyocytes via up-regulating GATA-4 expression, which enhanced interactions between Bcl-2 and Beclin-1.

Acknowledgements

This work was partly supported by grants from Guangdong Province Universities and Colleges Pearl River Scholar Funded Scheme (2016, A1-AFD018181Z3903 to Pro.Yongjun Chen), Innovation and Strengthen School Project of Guangdong Provincial Education Department-National Major Cultivation Project (2014GKXM031 to Pro.Nenggui Xu).


  A Preclinical Exploration of the Beneficial Effect of Tricin-Containing Medicinal Grain in Colon Health Top


Grace G. L. Yuea,b, Julia K. M. Leea,b, Si Gaoa,b, Yuk-Yu Chanc, Pang-Chui Shawa,b,c,d, Monique Simmondse, Clara B. S. Laua,b,c

aInstitute of Chinese Medicine, The Chinese University of Hong Kong,bState Key Laboratory of Research on Bioactivities and Clinical Applications of Medicinal Plants, The Chinese University of Hong Kong,cLi Dak Sum Yip Yio Chin R and D Centre for Chinese Medicine, The Chinese University of Hong Kong,dSchool of Life Sciences, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong, eRoyal Botanic Gardens, Richmond, United Kingdom

Correspondence:

Clara Bik-San Lau, E-mail: claralau@cuhk.edu.hk

Introduction: Colorectal cancer (CRC) is the third most frequently diagnosed cancer and has the second highest cancer mortality worldwide. Over 20 % of CRC patients present with relapse metastatic disease. Increasing evidences supported an association between healthy dietary patterns and decreased risk of CRC. Tricin, a flavone isolated from rice bran, has been previously shown to interfere with intestinal carcinogenesis in mice.[1],[2] In this study, we firstly demonstrated the presence of tricin in Chinese medicinal grain Oryzae Fructus Germinatus (OFG, germinated rice). Hence, our study aimed at evaluating the potential effects of tricin and OFG water extract on colon cancer metastasis in cell-based and tumor-bearing mice models.

Materials and Methods: The anti-migratory activities of tricin and OFG water extract were examined in two different human colon cancer cell lines. Besides, mice with orthotopic mouse colon-26 tumors received oral administration of tricin or OFG water extract, and tumor growth, metastasis in lungs, as well as tumor microenvironment were assessed after treatments.

Results: Tricin (7-14 mM) and OFG water extract (400-800 mg/mL) could significantly reduce the motility of human colon cancer cells in vitro. Both tricin (37.5 mg/kg) and OFG water extract (1500 mg/kg) treatments also suppressed the colon tumor growth and tumor cells metastasis to lungs. Furthermore, the populations of macrophages in tumors and myeloid-derived suppressor cells in spleens of tumor-bearing mice were modulated after tricin and OFG water extract treatments, respectively.

Conclusion: Oral administration of tricin or tricin-containing OFG water extract could suppress colon tumor growth and metastasis in mice. This is the first study showing the beneficial effect of the tricin-containing medicinal grain OFG in metastatic CRC.


  References Top


  1. Hudson EA, Dinh PA, Kokubun T, Simmonds MS, Gescher A. Characterization of potentially chemopreventive phenols in extracts of brown rice that inhibit the growth of human breast and colon cancer cells. Cancer Epidemiol Biomarkers Prev 2000;9:1163-70.
  2. Cai H, Al-Fayez M, Tunstall RG, Platton S, Greaves P, Steward WP, et al. The rice bran constituent tricin potently inhibits cyclooxygenase enzymes and interferes with intestinal carcinogenesis in ApcMin mice. Mol Cancer Ther 2005;4:1287-92.



  Natural Triterpenoid Actein Inhibits the Migration of Human Breast Cancer Cells in vitro and in vivo Top


Xiao-Xiao Wua, Grace G. L. Yueb,c, Ming-Hua Qiud, Clara B. S. Laub,c, Chun-Kwok Wonga,b,c

aDepartment of Chemical Pathology, The Chinese University of Hong Kong,bInstitute of Chinese Medicine, The Chinese University of Hong Kong,cState Key Laboratory of Research on Bioactivities and Clinical Applications of Medicinal Plants, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong SAR,dState Key Laboratory of Phytochemistry and Plant Resources in West China, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming, China

Correspondence:

Clara Bik-San Lau, E-mail: claralau@cuhk.edu.hk

Chun-Kwok Wong, E-mail: ck-wong@cuhk.edu.hk

Ming-Hua Qiu, E-mail: qiuminghua@mail.kib.ac.cn

Introduction: Breast cancer is currently the highly malignant cancer and causes the greatest number of cancer-related deaths in women. More than 90 % breast cancer death is due to metastasis. Anti-metastatic agents are urgently needed since standard chemotherapeutics cannot diminish the metastatic rate. Actein, a cycloartane triterpenoid, has been previously demonstrated to possess anti-angiogenic and anti-tumor activities.[1],[2] This study aimed to investigate the anti-migratory effect of actein and its underlying mechanisms of action.

Materials and Methods: In vitro effects on the proliferation, cell cycle phase distribution and migration were evaluated in two human breast cancer cell lines, MDA-MB-231 (ER negative) and MDA-MB-361 cells (ER positive). Western blots were employed to examine the protein expression of relevant regulatory molecules. In vivo effect of actein was examined using a human metastatic breast cancer cell (MDA-MB-231) xenograft in transparent zebrafish embryos model.

Results: In vitro experiments showed that actein treatment (10-40 μM) significantly decreased cell proliferation, migration, and caused G1 phase cell cycle arrest, as well as suppressed the protein expression of matrix metalloproteinases (MMPs) in both cancer cells. MDA-MB-231 cells were used to establish the human cancer cells xenograft in zebrafish embryos model since these cells are more prone to metastasis than MDA-MB-361 cells. In vivo results showed that actein at 60 μM could significantly decrease the number of zebrafish embryos with migrated cancer cells by 74 %.[3]

Conclusions: Actein exhibited both in vitro and in vivo anti-migratory activities. Therefore, actein could be a potential anti-metastatic natural compound for breast cancer treatment.


  References Top


  1. Yue GG, Xie S, Lee JK, Kwok HF, Gao S, Nian Y, et al. New potential beneficial effects of actein, a triterpene glycoside isolated from Cimicifuga species, in breast cancer treatment. Sci Rep 2016;6:35263.
  2. Einbond LS, Shimizu M, Nuntanakorn P, Seter C, Cheng R, Jiang B, et al. Actein and a fraction of black cohosh potentiate antiproliferative effects of chemotherapy agents on human breast cancer cells. Planta Med 2006;72:1200-6.
  3. Wu XX, Yue GG, Dong JR, Lam CW, Wong CK, Qiu MH, et al. Actein inhibits the proliferation and adhesion of human breast cancer cells and suppresses migration in vivo. Front Pharmacol 2018;9:1466.



  The Level of Depression and Anxiety in Patients with Different Sub-Types of Irritable Bowel Syndrome: A Systematic Review and Meta-Analysis Top


Zhi-Chao Hua, Mei-Xuan Lib,c, Liang Yaoa, Yin-Shu Wangd, En-Kang Wangd, Jian-Ye Yuand, Feng-Yun Wange, Zhao-Xiang Biana, Linda L. D. Zhonga

aHong Kong Chinese Medicine Clinical Study Center, School of Chinese Medicine, Hong Kong Baptist University, Hong Kong SAR,bSchool of Public Health, Lanzhou University,cEvidence-Based Medicine Center, School of Basic Medical Sciences, Lanzhou University,dInstitute of Digestive Diseases, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai,eXiyuan Hospital, China Academy of Chinese Medicinal Sciences, Beijing, P. R. China

Correspondence:

Zhaoxiang Bian, E-mail: bianzxiang@hkbu.edu.hk

Linda LD Zhong, E-mail: ldzhong@hkbu.edu.hk

Introduction: Irritable bowel syndrome (IBS) is a prevalent and chronic disease worldwide. In this study, we aimed to investigate the difference of the level of depression or anxiety in patients with different sub-types of IBS; and define which subtype of IBS is more likely to be associated with depression or anxiety.

Methods: A systematic search was conducted in Pubmed, EMBASE and the Cochrane library from inception to April 30, 2018. Cross-sectional, cohort or case–control studies that investigated anxiety and/or depression level in different subtype of IBS patients measured at baseline or the same point were included. Data collection included the characteristics of individual studies and their outcomes. Two independent reviewers conducted all research processes strictly. All statistical analyses were conducted on the r software. This study is registered with Prospetro, number crd42019124174.

Results: 12 studies with a total of 2259 IBS participants were included. Our study revealed no significant difference was presented in the level of depression and anxiety among different IBS subtype patients. Five studies reported the incidence rate of depression and anxiety in IBS-subgroup patients. We did not conduct a meta-analysis due to the high heterogeneity and small study sample.

Conclusions: Based on the best available evidence, there is no difference in the level of depression and anxiety in patients with different sub-types of IBS. Further studies on this topic should be carried out.


  The Acrine, an Analogue of Caffeine, Protects against Parkinson's Disease through Regulation of Mitochondrial Sirt3 Signaling Pathway Top


Wen-Jun Duana, Chang Yana, Ting-Mei Wanga, Shan-Bing Lia, Yi-Fang Lia, Hiroshi Kuriharaa, Rong-Rong Hea

aGuangdong Province Key Laboratory of Pharmacodynamic Constituents of TCM and New Drugs Research, College of Pharmacy, Jinan University, Guangzhou, China

Correspondence:

Dr. Rong-Rong He, E-mail: rongronghe@jnu.edu.cn

Introduction: Oxidative stress and loss of dopaminergic neurons in the substantia nigra pars compacta are reported to be the fundamental causes of Parkinson's disease (PD), one of the most popular neurodegenerative disorder. Although there is no standard cure for PD so far, researchers found that the consumption of tea or coffee could relieve the risk or development of the disease.

Materials and Methods: Materials: MPP+, MPTP, 6-OHDA, selegiline, caffeine, theacrine. Methods: Behavioral tests (spontaneous activity test, pole test, swimming test, rotarod test, Catwalk gait analysis), HPLC-ECD analysis, ELISA assay, flow cytometry, Western blotting analysis, immunoprecipitation, immunohistochemistry, etc.

Results: Theacrine was found to reduce the loss of dopaminergic neurons and relieve the damages of behavioral performance in animal models of PD (6-OHDA-treated SD rats and MPTP-treated C57BL/6J mice). In the subsequent studies investigating the potential mechanism, data indicated that theacrine relieved oxidative stress and mitochondrial dysfunction in MPP+-treated SH-SY5Y cells. We found theacrine concentration-dependently regulated both the activation and the protein expression of Sirt3, a mitochondrial deacetylase, by in vitro experiment of deacetylation reaction and Western blotting of cell lysates, respectively. Overexpression and silencing of Sirt3 were applied, and the results supported that Sirt3 was involved in the process of theacrine reversing the activation of caspase3.

Conclusions: Theacrine relieves the damages of behavioral performance by mitigating the mitochondrial oxidative stress and the loss of dopaminergic neurons through upregulation of both the expression and the activation of Sirt3.

Acknowledgements

This work was supported by National Natural Science Foundation of China (No. 81873209 & No. 81622050) and National Key Research and Development Program of China (2017YFC1700404).


  The Blockage of BRCC-Dependent K63 Deubiquitination of NLRP3 Facilitates the Inhibitory Effect of Celastrol on NLRP3 Inflammasome Activation Top


Xi Wanga, Shu-Hua Ouyangb, Chang-Yu Yana,b, Kurihara Hiroshia,b, Yi-Fang Lia,b, Rong-Rong Hea

aGuangdong Engineering Research Center of Chinese Medicine and Disease Susceptibility, Jinan University,bInstitute of Traditional Chinese Medicine and Natural Products, College of Pharmacy, Jinan University, Guangzhou, China

Correspondence:

Yi-Fang Li, E-mail: liyifang706@jnu.edu.cn

Rong-Rong He, E-mail: rongronghe@jnu.edu.cn

Introduction: Celastrol, an active ingredient extracted from traditional Chinese medicine Tripterygium wilfordii, exhibits anti-inflammatory activities NF-κB pathway in a variety of inflammatory diseases. This study aims to investigate whether the inhibition of NLRP3 inflammasome is engaged in the anti-inflammatory activities of celastrol and delineate the mechanism.

Materials and Methods: The influence of celastrol on NLRP3 inflammasome activation was first studied in THP-1 and mouse bone marrow macrophages (BMDM) treated with LPS followed by nigericin. Reconstitued inflammasome was also established by co-transfecting NLRP3, ASC and pro-caspase1 in HEK293T cells. The changes of inflammasome components including NLRP3, ASC, pro-caspase1/caspase, and pro-IL-1β/IL-1β were examined by ELISA, western blotting and immunofluorescence. P. acnes/LPS-induced liver injury and MSU-induced gouty arthritis in mice were employed as in vivo evidences to validate the inhibitory effect and mechanism of celastrol on NLRP3 inflammasome.

Results: Celastrol significantly suppressed the cleavage of pro-caspase1 and pro-IL-1β, while not affecting the protein expressions of NLRP3, ASC, pro-caspase1 and pro-IL-1β in LPS plus nigericin treated THP-1 and BMDM, and reconstituted inflammasome system in HEK293T cells. Similar results were obtained from in vivo experiments. Mechanism study revealed that celastrol could interdict BRCC-dependent K63 deubiquitination of NLRP3 which is critical in NLRP3 activation, thus prohibiting the formation of NLRP3, ASC and pro-caspase1 complex to block the generation of mature IL-1β.

Conclusions: Celastrol inhibits the K63 deubiquitination of NLRP3, thereby impeding the activation of NLRP3 inflammasome, which provides more evidences for the application in the therapy of inflammation-related diseases.

Acknowledgements

This work was supported by Youth Top-notch Talent Support Program of Guangdong Province (2016TQ03R586), and Program of Hong Kong Scholar (XJ2016017).


  Challenges and Prospects in Evaluating Outcomes Based on Traditional Chinese Medicine Symptoms Scoring System in Clinical Studies Top


Yuan Chia, Jian-Ping Liua

aCentre for Evidence-Based Chinese Medicine, Beijing University of Chinese Medicine, Beijing, China

Correspondence:

Jianping Liu, E-mail: jianping_l@hotmail.com

Introduction: ICD-11 has included traditional Chinese medicine (TCM) terms, indicating an international acceptance of TCM diagnostics. However, it remains problematic when including TCM symptoms score into outcome assessment in systematic reviews, as the standard of scoring in the primary studies varies from each other. This review revealed its challenges and prospects.

Materials and Methods: Literature review.

Results: Since 1978, China Ministry of Health issued guiding principles for clinical research including TCM symptoms scoring system, which grades each symptom of diseases quantificationally into slight, moderate and severe with values. An overall score and scores for each symptom then present a multidimensional evaluation for a disease. To produce reliable quantitative gradings and instruments of the symptom score, Chinese medical association and communities have conducted consensus meetings and evaluation on the validity and efficacy. Concerns over the evaluation of TCM symptoms score mainly related to the repeatability of measurement results and weight assignment in calculation among multiple symptoms. As for the repeatability, both patients' understanding of terminology for self-measurement items and clinicians' the knowledge and experience in diagnosing the symptoms affect the results simultaneously. For multiple symptoms, debate over the number, percentage, weight and value of each primary symptom and accompanied ones lowered the scientific nature of the scoring.

Conclusions: TCM symptoms scoring system necessarily requires a standardized way of presenting. And once verified by reliability and validity, it should be translated into English in time. This could contribute positively to WHO's concept of universal health care and the establish of TCM context-based efficacy evaluation.


  Refined Shuanghuanglian against CT26 Tumor Cells Seeding in Chronic Inflammatory Microenvironment of Lung Tissue Top


Cheng Lia, Pei-Huang Weia, Nuo Lia, Bo-Wen Weia, Jian Lia

aSchool of Traditional Chinese Medicine, Beijing University of Chinese Medicine, Beijing, China

Correspondence:

Jian Li, E-mail: lijian@bucm.edu.cn

Introduction: Paget suggests that tumor cells (” seeds “) selectively seek target organs (fertile “soil”) suitable for colonization and metastasis, but how the target organs affects metastatic seeding and growing still largely unknown. Many studies have shown that chronic inflammation enhanced the growth of metastatic tumors in mice and human.[1],[2] Improve the inflammatory microenvironment of target organs could reduce tumor risk, delay tumor progress, and reduce the burden of metastasis. Refined Shuanghuanglian (RSHL), composed of chlorogenica acid, baicalin and forsythoside A, could improve the inflammation of lung and has shown potential antitumor activity. Therefore, we attempted to observe the RSHL against metastatic seeding of CT26 colon tumour cell in chronic inflammatory microenvironment of lung tissue.

Materials and Methods: The BALB/c mice were intraperitoneally with RSHL (60 mg/kg/day) after intratracheal injected with bleomycin for 3 days, and CT26 cells were injected into the tail vein. The lungs were harvested after 1 week. The lung homogenates were subjected to ProcartaPlex M Th1/Th2 17-Plex Each analysis. The metastasis burden of lung tissues were analyzed by hematoxylin-eosin staining. The cell proliferation, migration and apoptosis in vitro were evaluate by CCK-8 assay, wound-healing assay and flow cytometry.

Results: The structural formula of RSHL [Figure 1]a. RSHL (50 μM) can inhibit the proliferation, migration and apoptosis of CT26 in vitro [Figure 1]b. Chronic inflammatory microenvironment of lung significantly promoted CT26 metastasis seeding compared with the model control group, and RSHL treatment improved it [Figure 1]c. Moreover, CT26 metastasis with bleomycin-treated decreased IL-27 and IL-2, and increased IL-13 and IL-22 in lung homogenate, while RSHL reversed it [Figure 2].
Figure 1: The antitumor activity of SHL in vitro and in vivo

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Figure 2: The expression of IL-27, IL-2, IL-13 and IL-22 in lung homogenate

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Conclusions: RSHL could against CT26 cell metastatic seeding in chronic inflammatory microenvironment of lung tissue.


  References Top


  1. Okada F. Inflammation-Related Carcinogenesis: Current Findings in Epidemiological Trends, Causes and Mechanisms[J]. Yonago acta medica 2014;57:65-72.
  2. Crusz SM, Balkwill FR. Inflammation and cancer: Advances and new agents[J]. Nature Reviews Clinical Oncology 2015;12:584-96.



  Systematic Study on TCM Status Naming of Type 2 Diabetes Top


Jing-Hui Zenga, Shi-Ping Chenga, Qing-Lin Zhaa

aJiangxi University of Traditional Chinese Medicine, Nanchang City, Jiangxi, P. R. China

Correspondence:

Zha Qinglin, E-mail: qinglin_zha@126.com

Introduction (Includes the Aim of the Study): It shows that different syndromes standards will have different Chinese Medicine pattern and pattern names via citing the syndromes standards in the published clinical research papers of Chinese medicine on type 2 diabetes. Therefore, this paper rise a systematic classification method of TCM state and standardized naming rules through the systematic analysis of TCM syndrome standard of type 2 diabetes.

Materials and Methods: The paper subjects on the cited Chinese Medicine syndromes standards in clinical research papers of type 2 diabetes, divides the name of syndromes into one or more basic syndromes units (abbreviated as state base), and the state base is divided into state elements (abbreviated as state element) of Chinese Medicine, which is composed by disease location elements and pathogenic elements, established a systematic classification system of TCM state of type 2 diabetes by using database and statistical analysis method, so as to determine the connotation of TCM state.

Results: We collected 49 syndrome standards and 407 syndrome names of diabetes, which covered 522 references, with a literature coverage rate of 91.1%. The result of disease location: (1) Pre diabetes: spleen 11, liver 4. (2) Diabetes period: stomach 47, lung 29, spleen 22. (3) Complication period: kidney 7, liver 5, spleen 2. The result of pathogenic factors: (1) Pre diabetes: Qi stagnation 8, phlegm obstruction 4, dampness (body fluid retention) 4, Yin deficiency 9, Qi deficiency 5. (2) Diabetes period: Yin deficiency 103, Qi deficiency 49, Yang deficiency 43, heat (Yang predominance) 59, excess heat (accumulated heat) 19, depression heat 15, blood stasis 18, phlegm 10, qi stagnation 9. (3) Complication period: Yin deficiency 15, Yang deficiency 9.

Conclusions: This paper systematically summarizes the Chinese Medicine syndrome standards of diabetes and puts forward the naming rules and classification methods of Chinese Medicine state system of type 2 diabetes based on location elements and pathogenic elements.


  Lable-Free Based Proteomics Method to Analyze the Mechanism of Components from Pulsatilla chinensis Saponins on NCI-H460 Lung Cancer Cells Top


Lan-Ying Chena, Zi-Yi Guana, Yi-Han Zhoua, Ying-Ying Luoa, Ya-Ru Cuia, Bin-Yao Shoua

aNational Pharmaceutical Engineering Center for Solid Preparation of Chinese Herbal Medicine, Jiangxi University of Traditional Chinese Medicine, Nanchang, China

Correspondence:

Lan-Ying Chen, E-mail: clyxy2513@163.com

Introduction: This study was to investigate the anti-tumor mechanism of PSD, R13 and PSA from Pulsatilla chinensis saponins components.

Materials and Methods: The inhibitory rate of PSD, R13 and PSA. on NCI-H460 Lung Cancer cell by MTT. The synergistic effects of three components were study by Calcusyn3.0 software. Apoptosis was analyzed by flow cytometry and DAPI stain. The molecular mechanisms was investigated by a proteomics approach combined with Nano-LC-MS/MS. Proteins exhibiting differential expression were analyzed using DAVID Bioinformatics Resources and KEGG databases. The differentially-expressed proteins were verified using western blotting.

Results: PSD, R13 and PSA have the synergistic anti-tumor effect were confirmed by Calcusyn3.0 software. Bioinformatics analysis showed that the overviews of the 116 differentially expressed proteins in the biological process (BP), cell component (CC), and molecular function (MF) and KEGG pathways categories. Some candidate proteins involved in carbon metastasis processes and PI3K/AKT/mTOR signaling pathways to induce apoptosis, which were validated by western blotting on NCI-H460 cell.

Conclusions: The anticancer synergistically mechanism of PSD, R13 and PSA may involve intervention in the carbon metastasis processes and PI3K/AKT/mTOR signaling pathways to induce apoptosis.

Acknowledgements

This work was supported by National Natural Science Foundation of China (No. 81460618; No. 81860720).


  The Energy Metabolism Mechanism of Multi-Components from Pulsatilla Chinensis Saponins Synergistically on NCI-H460 Top


Lan-Ying Chena, Zi-Yi Guana, Yi-Han Zhoua, Ying-Ying Luoa, Ya-Ru Cuia, Bin-Yao Shoua

aNational Pharmaceutical Engineering Center for Solid Preparation of Chinese Herbal Medicine, Jiangxi University of Traditional Chinese Medicine, Nanchang, China

Correspondence:

Lan-Ying Chen, E-mail: clyxy2513@163.com

The study was designed to investigate the anti-tumor effect of Pulsatilla chinensis saponins components and the synergistic effect of its effect components on tumor energy metabolism. NCI-H460 cell lines were cultured in vitro, MTT assay was used to detect the inhibitory rate of various components of Pulsatilla saponins. The compatibility and synergistic anti-tumor effect of different effect components were study by Calcusyn3.0 software. Biochemical assay and Elisa assay were used to detect the energy metabolism related metabolites (pyruvate, lactic acid, glucose) and enzymes (GLUT1, HK, PKM2, LDHA). Energy metabolism related gene members from KEGG pathway database and key protein (ERK1/2, RAS, GLUT1, MCT4) were detected by Real-time PCR and Western blotting.

The anti-tumor results in vitro showed that PSD, R13 and PSA in Pulsatilla chinensis saponins have the strongest anti-tumor activity for NCI-H460 human lung cancer cells and the IC50 were 5.2 ug·mL-1, 4.6 ug·mL-1 and 7.9 ug·mL-1, respectively. Using Calcusyn3.0 software to confirm the distribution ratio of three effective monomers in Pulsatilla chinensis saponins and determine the synergistic anti-tumor effect after compatibility. The results of biochemical and Elisa assay showed that the contents of pyruvate, lacate, glucose, HK, PKM2 and LDHA were significantly decreased and the GLUT1 content was significantly increased compared with the blank group (p < 0.05), compared with each monomer group, the content of pyruvic acid, lactic acid, glucose, HK, PKM2 and LDHA in the combined group were significantly decreased and the content of GLUT1 was significantly increased (p < 0.05). Real-time PCR results show that the combinations group has the most nodes in the network diagram, and its target is more than each monomer group. Western blotting results showed that compared with the blank group, the combinations group can significantly decreased the expression of ERK1/2, RAS, GLUT1, MCT4 protein (p <0.05). The combination of effect components of Pulsatilla chinensis saponins has synergistic anti-tumor effect on NCI-H460 and the anti-tumor mechanism may be associated with regulating energy metabolism.


  Systematic Study of TCM Status of Senile Diseases Top


Lan-Lan Chena, Miao Jiangb, Se-Qi Lina

aJiangxi University of Traditional Chinese Medicine, Nanchang City, Jiangxi,bInstitute of Basic Research in Clinical Medicine, China Academy of Chinese Medical Sciences, Dongcheng District, Beijing, P. R. China

Correspondence:

Lin Seqi, E-mail: lsq2517@163.com

Introduction (Includes the Aim of the Study): Systematically analyse the Chinese Medicine syndromes in the metadata of modern Chinese Medicine literature on the current senile diseases. And to explore the systematic and standardized classification methods of Chinese Medicine state of senile diseases, so as to understand the characteristics of Chinese Medicine state of senile diseases.

Materials and Methods: The modern Chinese Medicine clinical literature metadata of senile diseases was taken as the research object. Extracted the Chinese Medicine syndrome term in the subject or keyword, and divided the syndrome name into one or more basic syndrome units (referred to as the basic state); The basic state was divided into Chinese Medicine state elements (referred to as state element), which was composed of the location of disease element and the pathogenic and pathogenesis element; Then systematically classify and encode the elements. Based on these, the characteristics of Chinese Medicine state of senile diseases were analysed.

Results: 8,822 clinical articles on modern Chinese medicine for the treatment of senile diseases were collected. 1099 articles with subjects or keywords containing syndrome names, and there are 322 non-repeated Chinese Medicine syndrome names. In the Chinese Medicine state of senile diseases, the main on the location of disease elements were: no specific location of disease accounted for 56.36%, kidney 17.91%, liver 6.73%, spleen 6.16%, lung 5.46%, heart 1.75%, stomach 1.31%. The main the pathogenic and pathogenesis elements were: blood stasis 11.64%, sputum resistance 3.91%, qi stagnation 3.51%, yin deficiency 11.07%, qi deficiency 10.32%, yang deficiency 5.88%.

Conclusions: This paper proposed a naming rule of Chinese Medicine state based on the location of disease element and the pathogenic and pathogenesis element. Constructed the system classification method of Chinese Medicine state, and the characteristics of the Chinese Medicine state of the elderly in the metadata of modern Chinese Medicine clinical literature were summarized systematically.


  Pulsatilla Saponin Reduces Tumor Growth and Metastasis by Inhibiting M2 Macrophages Polarization via IL-4/STAT6 Pathway Top


Miao-Lin Wua, Xin Yana, Lan-Ying Chena, Jun Yua, Ying-Ying Luoa

aJiangXi University of Traditional Chinese Medicine, Nanchang, China

Correspondence:

Yingying Luo, E-mail: luoying0302@163.com

Xin Yan, E-mail: yanxinandy@163.com

LanYing Chen, E-mail: clyxy2513@163.com

Introduction: To explore the molecular mechanism of anti-tumor effect of Pulsatilla saponin by inhibiting M2 macrophage polarization.

Materials and Methods: Bone marrow-derived macrophages (BMDM) were pretreated with Pulsatilla saponin and stimulated by IL-4 to M2 macrophages. Related markers were detected by qRT-PCR to explore the effect of Pulsatilla saponin on M2 macrophages polarization;Conditioanl medium was collected ,The effect of Pulsatilla saponin on B16-F0 cells proliferation was examined by MTT assay, and Scratch test was used to examine the migration of B16-F0 melanoma ;Western Blotting was used to detect the phosphorylation of STAT6 in M2 macrophage. All above assays were repeated after interventing with STAT6 inhibitor AS1517499.

Results: qRT-PCR results showed Pulsatilla saponin significantly down-regulated the expression level of M2 macrophage marker genes Arg1, Fizz1, Ym1, and CD206. MTT assay and scratch test results showed that Pulsatilla saponin can significantly slow down the proliferation and migration of B16-F0 cells by affecting the polarization of M2 macrophages. 3 Western Blot assay showed that compared with vehicle (IL-4), the Pulsatilla saponin group significantly down-regulat the phosphorylation level of STAT6. The inhibition effect on macrophage polarization of Pulsatilla saponin + inhibitor AS1517499 group did not change,Compared with the group of Pulsatilla saponin. The same results were obtained from MTT assay,scratch test,and blotting p-Stat6.

Conclusions: Pulsatilla saponin reduces tumor growth and metastasis by inhibiting M2 macrophages polarization via IL-4/STAT6 pathway.


  Development of Pyrrole-Hemoglobin Adducts as a Biomarker for Monitoring Pyrrolizidine Alkaloid-Induced Liver Injury Top


Ge Lina, Ma Jianga

aSchool of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Hong Kong, China

Pyrrolizidine alkaloids (PAs) are phytotoxins widely distributed in ~3% of flowering plants. PA-induced liver injury (PA-ILI) is caused by metabolic activation of PAs by hepatic P450 enzymes to generate reactive pyrrolic metabolites resulting in formation of pyrrole-protein adducts, which impair the normal function of critical proteins in liver leading to hepatotoxicity. Our previous studies have illustrated the potential of blood pyrrole-protein adducts severing as a biomarker for monitoring PA exposure. The present study aimed further characterizing biochemical properties of blood pyrrole-protein adducts, including binding preference of reactive pyrrolic metabolites to different blood proteins, and examining elimination kinetics of different blood pyrrole-protein adducts, to identify a better biomarker (with higher level and longer half-life in bloodstream) for monitoring PA exposure and PA-ILI. Using our previously established pre-column derivatization followed by UHPLC-MS analysis, we quantified pyrrole-protein adducts in monocrotaline (a representative toxic PA)-treated rat blood protein fractions, including hemoglobin (Hb), plasma, albumin, and plasma residual protein fractions, and found that amount of pyrrole-Hb adducts was significantly higher than that in all plasma fractions. Moreover, elimination half-life of pyrrole-Hb adducts was significantly longer than pyrrole-protein adducts in plasma fractions (12.08 vs 2.54–2.93 days). In addition, we tested blood samples obtained from PA-ILI patients and confirmed that amount of pyrrole-Hb adducts were also remarkably higher than that of pyrrole-plasma protein adducts. In conclusion, our findings demonstrated that pyrrole-Hb adducts with remarkably higher level and longer life span can server as a better biomarker for monitoring PA exposure and PA-ILI.

Acknowledgements

The study was supported by GRF Grants (Ref Nos.: 14110714 and 14111816), and CUHK Direct Grant (Ref No.: 4054428).


  Determination of Six Compound in Human Plasma after Oral Administration of GalGeun-Tang (葛根湯) by UPLC-MS/MS Top


SeonYu Leea, JiHyun Jeonga, JinHee Parka, GukYeo Leea

aNational Development Institute of Korean Medicine, Jangheung-Gun Jeollanam-Do, Republic of Korea

Correspondence:

GukYeo Lee, E-mail: nara8938@nikom.or.kr

Introduction: GalGeun-Tang (GGT, gegen-tang in china and kakkon-to in japan), a herbal medication used in traditional therapy. It is used in common cold, fevers, headaches. The present study was conducted to development the UPLC-MS/MS method for the simultaneous analysis of six components in human plasma after oral administration of GGT (7.68 g/Dosd).

Materials and Methods: GGT(standardized as Korean Health Insurance, tablet) was supplied by Kyoungbang Pharmaceutical Inc. (Incheon, Korea). Six bioactive components for quantitation were Ephedrine(EPH), Puerarin(PRR), Daidzin(DIZ), Albiflorin(AF), Paeoniflorin(PF), 18β-glycyrrhetinic acid(18β-GTA), Tolbutamide(internal standard). The LC-MS/MS equipment included and Aglient 1290 system and 6470 triple quadrupole mass spectrometer.

Results: Contents of EPH, PRR, DIZ, AF, PF and glycyrrhizin(GL, It is metabolized to 18β-GTA by in human intestinal bacteria.) in 7.68 g of the GGT were calculated as 47.0 mg, 47.2 mg, 14.0 mg, 50.4 mg, 74.4 mg and 46.9 mg, respectively. Pharmacokinetics parameters(AUC0-∞, Tmax and Cmax) were calculated using the analysis results of plasma.

Conclusions: We evaluated bioactive analytes and PKs of compounds after oral dose of GGT in human. In addition, The analysis method for quantification of six component in human plasma was presented in the study.

Acknowledgements

This poster was supported by a grant from the Modernization project of Korean medicinal preparations (Pharmacololgical Standardization), National Development Institute of Korean Medicine(NICOM) and by a grant from the ministry of health and walfare, Republic of Korea.


  Proposed Management Method of Non-Standard Medicinal Herbs in External Herbal Dispensaries Top


Angela-Dongmin Sunga, Kyeong-Han Kimb, Hee-Jung Leea, Ji-Won Kima, Ji-Yeon Ryua, Soo-Hyung Sunga

aDepartment of Korean Medicine Policy, National Development Institute of Korean Medicine, Jung-Gu, Seoul,bDepartment of Preventive Medicine, College of Korean Medicine, Woosuk University, Wanju County, Korea

Correspondence:

Soo-Hyun Sung, E-mail: koyote10010@nikom.or.kr

Background: The Korean government has implemented the standard medicinal herb regulation so that it can be managed in systematic way. However in contrary, there are limited basic data regarding the distribution and production of non-standard medicinal herbs. In the case of raw material medicine needed for prescribing medicinal herbs, due to the mandatory standard medicinal herb regulation, most of them are being produced at GMP manufacturing companies. However there are cases where non-standard products are being used due to certain practical reasons.

Aim: In the case of the examination of the present condition of non-standard medicinal herbs conducted in this study, the aim was to understand the overall status of production and manufacturing in all external herbal dispensaries and medicinal herb manufactures.

Methods: During the period of October24th 2018 until the 22nd of November, an email survey was conducted to 98 national external herbal dispensaries and 151 medicinal herbs manufacturers based on a structured survey.

Results: Based on the responses from 42 external herbal dispensaries(responsive rate comparative to previous studies : 42.9%) and 43 medicinal herbs manufactures (28.5%), a study was conducted by utillizing descriptive statistics and frequency analysis. A study needs to be conducted in future on the status of distribution and usage of non-standard medicinal herbs within the country, what the institutional problems are, and how aware agents are of this matter.


  Introduction of an Accreditation System for Controlling Quality of Herbal Medicine in Korea Top


Hee-Jung Leea, Angela-Dongmin Sunga, Ji-Won Kima, Ji-Yeon Ryua, Soo-Hyun Sunga

aDepartment of Korean Medicine Policy, National Development Institute of Korean Medicine, Jung-Gu, Seoul, Korea

Correspondence:

Soo-Hyun Sung, E-mail: koyote10010@nikom.or.kr

Objective: The objective of this study is to review effects of the introduction of an accreditation system of external herbal dispensaries (EHDs) that installed by traditional Korean medicine (TKM) clinics on the quality control of herbal medicines (HMs).

Background: In 2008, Korea Ministry of Health and Welfare (MHW) established guideline on installation and utilization of EHDs and share-use of EHDs. Since then, the number of EHDs has increased to 98 in 2017, and the proportions of HMs supplied to patients in EHDs have also increased. EHDs contract with the TKM clinics and supply HMs prepared by prescription to clinics.

Introduction of Accreditation System: Korea MHW introduced an autonomous evaluation and certification system in 2018 for quality control of EHD that prepare HMs. Evaluation standards were composed of 81 evaluation items including facilities, preparation environment, document management, employee management, management of herbs and preparation process.

Quality Control of Herbal Medicine: Herbal dispensaries have been responsible for preparing HMs that are used for treating patient's diseases. The accreditation of EHD is new system for controlling quality of herbal medicine not licensed by Korea food and drug administration (FDA).

Conclusions: The accreditation system of EHD will enhance the safety and quality of HMs.


  Effect of Soshiho-Tang on Cytochrome P450 Drug-Metabolizing Enzymes in Rat Top


Da Hwa Junga, Si Won Kanga, Seol U. Jia, Su-Jung Parka, Seong Ho Hama

aNational Development Institute of Korean Medicine, Janghueung-Gun, Jeollanam-Do, Republic of Korea

Correspondence:

Seong Ho Ham, E-mail: phd_ham@hanmail.net

Introduction: Cytochrome P450 (CYP) enzymes, which are primarily located in the liver, are responsibility of the first metabolism of all drugs. Measuring the degree of induction of CYP enzymes in drug candidates is crucial for drug interaction, drug efficacy, and safety assessment. The aim of this study was to investigate the Cytochrome P450-catalyzed reaction of induction and activity in repeated-dose of in Soshiho-tang (SSH) soft extract of the Standardized Korean Heath Insurance (KHI) is administered for 5 days repeated-dose of in rat.

Materials and Methods: Rat liver microsome (RLM) extracted from SSH (1, 6, and 18 times) repeatedly dosed with for 5 days in male rats. To confirm the expression level of hepatic metabolism enzyme (CYP450), Western blot test and HPLC analysis based on the US FDA draft guidance (2016) test method for CYP activity test (CYP inhibition test) were performed.

Results: In Western blot test, CYP2C9, CYP2D6, and CYP2E1 were significantly decreased following dose dependence. The activity of CYP2D6, and CYP2E1 were significantly decreased respectively. Western blot and CYP activity showed similar tendency of inhibit 2D6, 2E1.

Conclusions: Results indicated that basic information for proving the drug interaction with the CYP2D6 (substrate drug : propranol) and CYP2E1 (substrate drug : Chlozoxazone) by SSH. Furthermore, it is necessary to confirm the significance test for CYP2D6 inhibitor (Burpropion) and CYP2E1 (chlomethiazole) through comparative experiments.

Acknowledgements

This paer was supported grant from the Modernization Project of Korean Medicinal Preparation and the National Development Institue of Korean Medicine(NIKOM) and by a grant from Ministy of Health and Walfare, Republic of Korea.


  Hepatic Protective and Antioxidant Effects of Arecae pericarpium Extract against Alcoholic Liver Diseases Top


Sun-Gun Kima, Hye-Ryung Kanga, Hyun-Sook Kwona, Bo-Mi Kima, Ji-Eun Parka, Jai-Hyun Soa

aDepartment of Korean Medicine Development, National Development Institute of Korea Medicine, Gyeongsan-Si, Republic of Korea

Correspondence:

Sun-Gun Kim, E-mail: sungun@nikom.or.kr

Introduction: This study was carried out to evaluate the hepatoprotective and antioxidant effects of a Arecae pericarpium extract (APE) to relieve a hangover.

Materials and Methods: The change of alcohol contents (ADH, ALDH). Measurement of total antioxidant (DPPH, ABTS, FRAP, phenol).

Results: APE reduced the serum alcohol concentration to 27.1% in rat treated with alcohol. In accordance with this, APE improved ADH, ALDH, ALP, ALT, AST, TP and creatinine in the serum. In addition, APE showed strong ferric reducing activities and effectively scavenged radicals including DPPH, ABTS.

Conclusions: This results demonstrate that APE has the potential to reduce hangover and might be used as a functional food and natural medicine for preventing alcoholic liver diseases and its complications.

Acknowledgements

Hepatic protective, Antioxidant, Alcohol, Liver diseases, Hangover, Arecae pericarpium.


  The Gastroprotetive Effect of Leonuri Herba on HCl/Ethanol Induced Gastric Mucosa Damage in Rats Top


Mi-Ok Sima, Ji-Hun Janga, Ho-Kyung Junga, Jong Hyun Nhoa, Beodeul Yanga, Hyun-Woo Choa

aNational Development Institute of Korean Medicine, Jangheung-Gun, Republic of Korea

Correspondence:

Hyun-Woo Cho, E-mail: johw7@nikom.or.kr

Introduction: Gastritis is a major complication of gastrointestinal disease.[1] Leonuri Herba (TKM2-14) is used in folk medicine as the treatment of many diseases.[2] Therefore, this study examined the effects of TKM-14 on HCl/ethanol induced acute gastric ulceration.

Materials and Methods: SD rats were randomly divided into five different groups including the normal group, ulcer group, positive group (20mg/kg of omeprazole), and experimental groups (100mg/kg and 500mg/kg of TKM2-14). Morphological features of gastric tissue were observed by ulcer score, and H&E staining.

Results: Macroscopic and histological analyses showed TKM2-14 were significantly reduced to an extent that it allowed leukocytes penetration of the gastric walls compared with the ulcer group. In addition, an ulcer inhibition rate and prostaglandin E2 levels were increased in rats treated with TKM2-14. These results suggested that TKM2-14 reduced gastric ulcer and the protection of gastric wall, shown in [Figure 1].
Figure 1: Effect of TKM2-14 on rats

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Conclusions: The present study has demonstrated the gastroprotective effect of TKM2-14, these findings suggested that TKM2-14 has the potential for use in treatment of gastric disorders.

Acknowledgements

This work was supported by the Ministry of Health & Welfare Korea and Jellanamdo “Basic Standardization Project for Herbal Medicinal Materials of Korean Indigenous Resources”.


  References Top


  1. Al-Quraishy S, Othman MS, Dkhil MA, Abdel Moneim AE. Olive (Olea europaea) leaf methanolic extract prevents HCl/ethanol-induced gastritis in rats by attenuating inflammation and augmenting antioxidant enzyme activities. Biomed Pharmacother 2017;91:338-49.
  2. Liu XH, Pan LL, Zhu YZ. Active chemical compounds of traditional Chinese medicine Herba Leonuri: Implications for cardiovascular diseases. Clin Exp Pharmacol Physiol 2012;39:274-82.



  Effects of Artemisiae Argyi Herba on the of Atopic Dermartitis Induced by by 2,4-Dinitrochlorobenzene Top


Beodeul Yanga, Ho-Kyung Junga, Ji-Hun Janga, Mi-Ok Sima, Jong Hyun Nhoa, Hyun-Woo Choa

aNational Development Institute of Korean Medicine, Jangheung, Jeollanam-Do, Korea

Correspondence:

Hyun-Woo Cho, E-mail: johw7@nikom.or.kr

Introduction: Atopic dermatitis (AD) is chronic inflammatory skin disease and has skin problems such as scab, eczema and itching. It characterized inflammatory response as such immunoglobulin (Ig) E mediated hypersensitivity reactions. Artemisiae Argyi Herba has been used widely as a traditional herbal medicine for improvement of inflammatory in China, Japan and Korea. Based on above, the aim of the study was to confirm whether Artemisiae Argyi Herba effects to improve inflammatory skin in AD-induced mice by 2,4-Dinitrochlorobenzene(DNCB).

Materials and Methods: We investigated the effects of water extract of Artemisiae Argyi Herba(TKMII-20) on surface of dorsal skin and histopathology changes in AD mice by DNCB. Furthermore, the effects on Ievel of IgE and filaggrin were identified.

Results: Application with TKMII-20 improved skin surface and reduced clinical severity score. TKMII-20 prevented epidermal hyperplasia, hyperkeratosis and immune cell infiltration. Furthermore, the level of IgE and filaggrin were reduced by TKMII-20 in AD mice serum, shown in [Figure 1].
Figure 1: Effect of TKMII-20 on IgE in serum of AD mice

Click here to view


Conclusions: These result identify that TKMII-20 can suppress development of AD induced by DNCB in BALB/c mice. These result suggest that TKMII-20 was effective in the treament on AD and TKMII-20 would prevent AD.

Acknowledgements

This work was supported by the Ministry of Health & Welfare Korea and Jellanamdo “Basic Standardization Project for Herbal Medicinal Materials of Korean Indigenous Resources.”


  Improvement Effect of γ-Pyrone-3-O-β-D-(6-Galloyl)-Glucopyranoside Isolated from Euphorbia ebracteolata var. Coreana on Cisplatin-Induced Nephrotoxicity Models Top


Kyeong Wan Wooa, Jong-Hyun Nhoa, Hyun-Ju Leea, Ho Kyung Junga, Hyun Woo Choa

aNational Development Institute of Korean Medicine, Jangheung-gun, Republic of Korea

Correspondence:

Hyun Woo Cho, E-mail: johw7@nikon.or.kr

Introduction: The roots of E. ebracteolata var. coreana have been used in traditional medicine to treat neuropathic dermatitis and pulmonary tuberculosis in China, Japan, and Korea. To investigate the improvement effect of γ-Pyrone-3-O-β-D-(6-galloyl)-glucopyranoside (PGP) isolated from E. ebracteolata var. coreana on cisplatin-induced nephrotoxicity using an animal model (BALB/c mice) and HK-2 cells.

Materials and Methods: The phytochemical investigation of 100 % MeOH extract of E. ebracteolata var. coreana using column chromatography resulted in the isolation of one phenolic derivative (1). Structural elucidation of the compound 1 was based on spectroscopic methods. The protective effects of PGP on cisplatin-induced HK-2 cell death were examined in vitro. Animals were divided into 5 groups and the beneficial effects of PGP on a cisplatin-induced nephrotoxicity animal model were analyzed by histological and hematological analysis in vivo.

Results: 80 μM PGP significantly reduced 20 μM cisplatin-induced apoptotic cell death in HK-2 cells through inhibition of mitochondrial dysfunction, DNA fragmentation, and caspase-3 activation. Moreover, renal dysfunction, tubular damage, and neutrophilia induced by 20 mg/kg cisplatin injection were significantly decreased by PGP at 10 mg/kg.

Conclusions: This study indicated that PGP treatment improved cisplatin-induced nephrotoxicity in vitro and in vivo, and could be used to improve cisplatin-induced adverse effects.


  Chemical Constituents of Dicentra spectabilis and their Anti-Inflammation Effect Top


A. Hyeon Kima, Ji Kyung Kima, Kyeong Wan Wooa, Ho Parka, Hyun Ju Leea, Ho Kyung Junga, Hyun Woo Choa

aNational Development Institute of Korean Medicine, Jangheung-gun, Republic of Korea

Correspondence:

Hyun Woo Cho, E-mail: johw7@nikon.or.kr

Introduction: Inflammation is an essential aspect of the host response to infection and injury and is a key factor for the maintenance of good health in response to infections. However, aberrant inflammation underlies many acute and chronic human diseases such as arthritis, and autoimmune disease The roots of D. spectabilis have been used for the treatment of various conditions such as strokes, bruises, and blood circulation. Previous phytochemical investigations on D. spectabilis have reported the isolation of fungitoxic alkaloids and compounds with apoptosis-inducing activities. However, only a few phytochemical studies on D. spectabilis have been reported. As part of the search for bioactive constituents of Korean medicinal plants, we investigated the active constituents of D. spectabilis.

Materials and Methods: Repeated column chromatographic separations of a MeOH extract led to the isolation of fourteen known compounds (1-14). All compounds were measured for their anti-inflammatory effects on a lipopolysaccharide (LPS)-induced murine microphage cell line RAW 264.7.

Results: D. spectabilis derived compounds 3, 4, and 12 repressed the production of these macrophage mediators in LPS-induced RAW264.7 cells. Compound 3, in particular, effectively inhibited LPS-induced pro-inflammatory cytokines (TNF-α, IL-6, and IL-1β).

Conclusions: The results suggest that these compounds could be used as anti-inflammatory drugs and substrates.


  Phytochemical Constituents from the Rhizomes of Osmunda japonica Thunb and their Anti-Oxidant Activity Top


Ji Kyung Kima, Ho Parka, A. Hyeon Kima, Hyun Ju Leea, Kyeong Wan Wooa, Ho Kyung Junga, Hyun Woo Choa

aNational Development Institute of Korean Medicine, Jangheung-gun, Republic of Korea

Correspondence:

Hyun Woo Cho, E-mail: johw7@nikon.or.kr

Introduction: Osmunda japonica (Osmundaceae) is a perennial herb, widely distributed throughout Taiwan, Japan, and Korea. Its rhizomes have long been used in traditional Korean medicine for treating hemostasis and fever. Previous phytochemical investigations of this plant resulted in the isolation of flavonoids and phenolic constituents. Anti-oxidant, anti-microbial and herbicidal effects of an MeOH extract of O. japonica have also been reported. As part of our ongoing research for bioactive compounds from indigenous plant in Korea, we investigated the MeOH extract of O. Japonica.

Materials and Methods: The rhizomes of O. japonica (2 kg) were collected at Wando-gun in Jeollanam-Do province in July 2015. A voucher specimen (TKM-2081) of the plant was deposited in the herbarium at NIKOM, Jangheung, Korea. Purification of the H2O-soluble fractions by multiple chromatographic steps led to the isolation of eleven known compounds (1-11) and their structures were elucidated based on1H,13C-NMR and LC-IT-TOF/MS data. The antioxidant activities of isolated compounds (1–11) were measured by DPPH and ABTS assays.

Results: Column chromatographic separation of aqueous fractions of A. alpina led to the isolation of 17 compounds. To the best of our knowledge, the present study is the first to identify compounds 2-10 and 12-17 in A. alpina. Furthermore, compound 6 possessed powerful antioxidant activity, while compound 15 suppressed intracellular tyrosinase activity and thus reduced melanogenesis in B16F10 cells.

Conclusions: A Korean indigenous plant O. japonica may be considered to be functional and effective material for treatment of various diseases.


  Antioxidant and Anti-Melanogenic Activities of Compounds Isolated from the Aerial Parts of Achillea alpina Top


Ho Parka, Ji Kyung Kima, A. Hyeon Kima, Hyun Ju Leea, Kyeong Wan Wooa, Ho Kyung Junga, Hyun Woo Choa

aNational Development Institute of Korean Medicine, Jangheung-gun, Republic of Korea

Correspondence:

Hyun Woo Cho, E-mail: johw7@nikon.or.kr

Introduction: Achillea alpina is widely distributed in Korea and is often used as a folk medicine for stomach disorders. Although a previous study isolated antioxidant compounds (flavonoid O-glucoside, sesquiterpene) from this plant, no systematic study of its chemical constituents had been reported. The present study aimed to identify the phytochemicals present in a methanol extract of A. alpina, assess their potential antioxidant activities in vitro, and determine their effects on melanogenesis in B16F10 melanoma cells.

Materials and Methods: The present analysis of the bioactive constituents of Korean medicinal plants identified 17 known compounds (1-17) in a methanol extract of A. alpina. The structures of 1-17 were characterized using nuclear magnetic resonance (NMR) methods (1H and13C-NMR) and liquid chromatography coupled to high-resolution tandem mass spectrometry (LC IT-TOF/MS) All of the isolated compounds (1-17) were evaluated for their antioxidant effects, as well as their effects on melanogenesis in B16F10 melanoma cells.

Results: Column chromatographic separation of aqueous fractions of A. alpina led to the isolation of 17 compounds. To the best of our knowledge, the present study is the first to identify compounds 2-10 and 12-17 in A. alpina. Furthermore, compound 6 possessed powerful antioxidant activity, while compound 15 suppressed intracellular tyrosinase activity and thus reduced melanogenesis in B16F10 cells.

Conclusions: Our research suggested that these naturally occurring compounds have the potential to reduce oxidative stress and promote skin whitening.


  The Therapeutic Effect of Korean Herbal Rosacea Cream and Silymarin in LL-37-Induced Rosacea Mouse Model Top


Ho Kyung Junga, Ji Hun Janga, Mi Ok Sima, Byeong Kwan Ana, Beodeul Yanga, Hyun Woo Choa

aNational Development Institute of Korean Medicine, Jangheung-gun, Republic of Korea

Correspondence:

Hyun-Woo Cho, E-mail: johw7@nikom.or.kr

Introduction (Includes the Aim of the Study): Rosacea is a common inflammatory dermatologic condition that affects locally around face and eyes. Even though rosacea is not a life-threatening condition, as it involves the face, it can build emotional, social, and psychological stresses. Therefore, the selection of the proper therapeutic cream for each patient with rosacea is important. The present study was designed to investigate anti-inflammatory effects of the novel Korean herbal rosacea cream and silymarin in LL-37 induced rosacea mouse model.

Materials and Methods: We researched the effects of Korean herbal rosacea cream on histological changes and score in LL-37 induced rosacea mouse model. Also, the effects on level of myeloperoxidase assay was investigated.

Results: Injection of LL-37 peptide cause significant redness, redness area, immune cell infiltration and collapsed skin layer compared of non-treated (NOR) group. These conditions were concentration-dependently ameliorated following the administration of the three doses of silymarin (Sil) and Korean herbal rosacea cream (Co)-treated groups when compared with the LL-37 group.

Conclusions: These results indicated that silymarin, Korean herbal rosacea cream inhibit LL-37-induced rosacea characteristic, and might be a potential therapy against rosacea.

Acknowledgements

This work was supported by the Ministry of Health & Welfare Korea and Jellanamdo “Basic Standardization Project for Herbal Medicinal Materials of Korean Indigenous Resources.”


  Development of Korean Medicine Clinical Guideline for Non-Specific Chronic Low Back Pain Top


Dongwoo Nama, Yejin Honga, Hyewon Yeuma, Serom Jeona

aDepartment of Acupuncture and Moxibustion, College of Korean Medicine, Kyung Hee University, Seoul, South Korea

Correspondence:

Dongwoo Nam, E-mail: hanisanam@hanmail.net

Purpose: As science and medicine evolves, the average life span of mankind is rapidly being extended.

Acupuncture is one of the most popular treatments to the patients seeking complementary medicine. As patient population receiving acupuncture increases worldwide, needs for a standardized clinical guideline is growing. Among the many diseases treated by acupuncture, musculoskeletal disorders rank the top.

The objective of this study is to establish Korean Medicine clinical guideline for non-specific chronic low back pain.

Methods: A task force team to establish the guideline was composed. Literature review was done in order to search for evidence of safety and efficacy of acupuncture and other Korean Medicine treatments. A survey was done in order to find out how Korean medical doctors derive pattern identification for acupuncture and herbal prescriptions in treating non-specific chronic low back pain. Then, based on the results of literature review and survey, a conference meeting of experts was held. Through the Delphi method, a draft of the acupuncture clinical guideline for non-specific chronic low back pain was established. Now the review board, composed of experts of musculoskeletal disorders, public health, statistics, and representatives of patients are modifying the draft.

Results: Evidence of safety and efficacy of acupuncture treatment for musculoskeletal disorders was established. A standard or pattern identification was derived. A draft of Korean Medicine clinical guideline for non-specific chronic low back pain was established.

Conclusions: More rigorous, well designed and large scaled RCTs are in need to improve the quality and make modifications this clinical guideline.

Keywords: Clinical guideline, Korean medicine, non-specific chronic low back pain


  Quality Grading of Chinese Medicinal Materials: Five Herbs, a Pilot Study Top


Li-Xia Zhua, Jun Xua, Xiao-Ping Donga, Hu-Biao Chena, Zhong-Zhen Zhaoa

aSchool of Chinese Medicine, Hong Kong Baptist University, Kowloon, Hong Kong Special Administrative Region, China

Correspondence:

Zhongzhen Zhao, E-mail: zzzhao@hkbu.edu.hk

Introduction (Includes the Aim of the Study): Currently, the quality grading standards of Chinese medicinal materials (CMMs) are empirical; while it should reflect quality-based pricing, it lacks a theoretical basis. Therefore, it is of great significance to determine the correlation between the quality of CMMs and the factors involved in the empirical standards to provide a scientific basis for their quality classification, market standardization, and rational utilization.

Materials and Methods: Different sources of five herbs, namely Polygoni Multiflori Radix (PMR), Cinnamomi Cortex (CC), Scutellariae Radix (SR), Uncariae Ramulus Cum Uncis (URCU), and Achyranthis Bidentatae Radix (ABR), were collected. Laser microdissection combined with UHPLC-QTOF-MS/MS, UHPLC-QqQ-MS/MS, ICP-MS, and GC-MS were applied to characterize distribution patterns of secondary metabolites in different tissues of CMMs, contents of bioactive components, fingerprints, heavy metals, and pesticide residues.

Results: The factors involved in the empirical grading standards, including weight for PMR, thickness for CC, commercial grade and growth years for SR, and commercial specifications for URCU and ABR, had little influence on the quality of corresponding CMMs. On the other hand, the thickness of specific tissues, namely cork and xylem-core for PMR, phloem for CC and SR, phloem and cortex for URCU, and cork and central xylem for ABR, and their place of origin affected it greatly.

Conclusions: The current empirical quality grading standards of these five herbs may lack scientific rationality. Nevertheless, it is worth considering the place of origin and histochemical characteristics when establishing scientific and reasonable quality grading standards for CMMs.


  Issues in Developing Traditional Medicine Evidence-Based Clinical Practice Guidelines Top


Minjung Parka, Suran Kima, Jaesuk Leea, Hyunjung Honga, Jongwoo Kimb, Seokhee Jungb, Nankwen Kima

aNational Development Institute of Korean Medicine, Junggu,bCollege of Korean medicine, Kyunghee University, Dongdaemoongu, Seoul, Republic of Korea

Correspondence:

Namkwen Kim, E-mail: drkim@nikom.or.kr

Introduction: Korean government initiated the large-scale national project for standardization of Korean medicine from 2016 to 2021, which includes development of 30 evidence-based traditional medicine clinical practice guidelines and conducting clinical trials for strengthening the evidence level of CPGs. There were some methodological issues requiring expert consensus in the course of developing 30 clinical practice guidelines simultaneously.

Materials and Methods: Guideline center for Korean medicine draw up issues in the several 30-CPG developers' meetings and conducted comprehensive prior literature review regarding to the issues. Then two different surveys went on to all 30 primary CPG researchers and to lay Korean medicine doctors as CPG's main potential users. Focus group interview was conducted with CPG methodology professionals who referred the result of literature review and surveys.

Results: The first issues was where to search to find comprehensive evidence related to Korean medicine. And second one was how to reflect the traditional theory and experiences. The third, the appropriate methodology to overcome the heterogeneity of intervention. As result of expert consensus, we added several databases from countries sharing similar theory and experiences with Korean medicine. We recommended modified level of recommendation reflecting characteristics of traditional medicine. Besides, we suggested the judgement criteria to overcome heterogeneity of intervention.

Conclusions: The guidelines should be developed taking into account the features of specific area and regional characteristics.


  Effect of Gumiganghwal-Tang on Cytochrome P450 Drug-Metabolizing Enzymes in Rat Top


Seol U. Jia, Da Hwa Junga, Si Won Kanga, Su-Jung Parka, Seong Ho Hama

aNational Development Institute of Korean Medicine, Jangheung-Gun, Jeollanam-Do, Republic of Korea

Correspondence:

Seong Ho Ham, E-mail: phd_ham@hanmail.net

Introduction: Cytochrome P450 (CYP) enzymes, which are primarily located in the liver, are responsibility of the first metabolism of all drugs. Measuring the degree of induction of CYP enzymes in drug candidates is crucial for drug interaction, drug efficacy, and safety assessment. The aim of this study was to investigate the Cytochrome P450-catalyzed reaction of induction and activity in repeated-dose of in Gumiganghwal-tang (GMGHT) soft extract of the Standardized Korean Heath Insurance (KHI) is oral-administered for 5 days repeated-dose of in rat.

Materials and Methods: Rat liver microsome (RLM) extracted from GMGHT (1, 6, and 18 times) repeatedly dosed with for 5 days in male rats. To confirm the expression level of hepatic metabolism enzyme (CYP450), Western blot test and HPLC analysis based on the US FDA draft guidance (2016) test method for CYP activity test (CYP inhibition test) were performed.

Results: In Western blot test, CYP2C19, CYP2D6, and CYP3A4 were significantly decreased following dose dependence. The activity of CYP2B6, CYP2D6 and CYP2E1 were significantly decreased, respectively. Western blot and CYP activity showed similar tendency of inhibit 2D6.

Conclusions: Results indicated that basic information for proving the drug interaction with the CYP2D6 (substrate drug: propranolol) by GMGHT. Furthermore, it is necessary to confirm the significance test for CYP2D6 inhibitor (Bupropion) through comparative experiments.

Acknowledgements

This paper was supported grant from the Modernization Project of Korean Medicinal Preparation and the National Development Institute of Korean Medicine (NIKOM) and by a grant from Ministry of Health and Welfare, Republic of Korea.


  Determination of Human Plasma Protein Binding for Fourteen Components in Gumiganghwal-Tang using UPLC-MS/MS Analytical Method Top


Seung-Hyun Jeonga, Ji-Hun Janga, Guk-Yeo Leeb, Seung-Jung Yangc, Hea-Young Chod, Yong-Bok Leea

aCollege of Pharmacy, Chonnam National University, Gwangju,bNational Development Institute of Korean Medicine, Jangheung-gun, Jeollanam-Do,cDepartment of Oriental Rehabilitation Medicine, Dongshin University Suncheon Medical Center, Suncheon-Si,dCollege of Pharmacy, CHA University, Seongnam-Si, Gyeonggi-Do, Korea

Correspondence:

Yong-Bok Lee, E-mail: leeyb@chonnam.ac.kr

Introduction: The aim of this study was to determine binding of Gumiganghwal-tang (GGHT) components to human plasma protein (HPP) in vitro and in vivo using a newly developed analytical method.

Materials and Methods: To determine the binding ratio to HPP in vitro, each 14 component of GGHT was added to blank human plasma to concentration of 0.1 or 1 μg/mL. Human plasma samples were obtained at 1 and 8 h after oral administration of GGHT tablets to determine the binding ratio to HPP in vivo. All samples were fractionated by ultrafiltration method using a centrifugal filter unit. Both the ultrafiltrates and the plasma samples without ultrafiltration were analyzed using the newly developed UPLC-MS/MS method. Finally, the percentage of HPP binding was calculated as (total drug concentration-ultrafiltrate drug concentration)/total drug concentration.

Results: In vitro average HPP binding ratios of 14 components of GGHT were 5.52-96.53%, which varied widely among the components. In vivo HPP binding ratios of components except for 18β-glycyrrhetinic acid, oxypeucedanin, atractylenolide III, baicalin, baicalein, and ferulic acid were not available due to not be able to detect them in clinical samples. However, the in vitro results and the average HPP binding ratios for the six components that could be countable in vivo were similar. Furthermore, our analytical method was simple, rapid, accurate, and sensitive for the component quantification of GGHT.

Conclusions: The developed analytical method could be successfully applied to in vitro and in vivo HPP binding studies of fourteen components. HPP binding of components is an important factor in pharmacokinetics as well as a factor that may have a significant effect on component efficacy.

Acknowledgements

This work was supported by a grant of the NIKOM funded by the Korean Ministry of Health and Welfare, Korea.


  Developing Clinical Practice Guideline of Korean Cultural Syndrome Hwabyung Top


Hyo-Weon Suha, Jong Woo Kima, Sun-Yong Chunga

aDepartment of Neuropsychiatry, Kyung Hee University Korean Medicine Hospital at Gangdong, Gangdong-Gu, Seoul, Republic of Korea

Correspondence:

Sun-Yong Chung, E-mail: lovepwr@khu.ac.kr

Introduction: Hwabyung, listed as culture-bound syndrome in The Diagnostic and Statistical Manual of Mental Disorders (DSM-IV), is an anger syndrome related to Korean culture. Korean medicine interventions are effective in treating hwabyung. Thus, we developed evidence-based clinical practice guideline for Korean medicine on hwabyung.

Materials and Methods: Working group selected the core clinical questions and conducted systematic reviews and meta-analyses. The level of evidence and recommendation grade were evaluated in accordance with GRADE (Grading of Recommendations Assessment, Development, and Evaluation), and the actual implementation process was referred to Korea Institute of Oriental Medicine, “Clinical Practice Guideline Development Project (K15080)”. The level of evidence and grade of recommendation were formulated through the Delphi technique.

Results: The recommendations were following: (1) Herbal medicine should be considered to treat hwabyung (B/Moderate). (2) Acupuncture should be considered to treat hwabyung (B/Low). (3) Pharmacopuncture at CV17 and GB21 is recommended to treat physical symptoms by clinical experience of this development group (GPP/No study). (4) Moxibustion at CV12, CV04, CV06, ST36, and LR03 is recommended to treat gastrointestinal symptoms such as dyspepsia by clinical experience of this development group (GPP/No study). (5) Cupping therapy at Governor Meridian and Gallbladder Meridian is recommended to treat muscular pain, insomnia, stuffy chest, and psychological tension by clinical experience of this development group (GPP / No study). (6) Emotional Freedom Techniques should be considered to treat hwabyung (B/Moderate). (7) Group counseling should be considered to treat hwabyung (B/Moderate). (8) Meditation should be considered to treat hwabyung (B/Moderate).

Acknowledgements

This study was supported by the Traditional Korean Medicine R&D program funded by the Ministry of Health & Welfare through the Korean Health Industry Development Institute (KHIDI) (HB16C0068).


  Chemical Stability of Antioxidative Principal-Acteoside from Flower Extracts of Osmanthus fragrans Lour Top


Hong-Yu Chena, Lih-Geeng Chena

aDepartment of Microbiology, Immunology and Biopharmaceuticals, National Chiayi University, Chiayi, Taiwan

Correspondence:

Lih-Geeng Chen, E-mail: lgchen@maiul.ncyu.edu.tw

Introduction: The flowers of Osmanthus fragrans Lour. (Oleaceae) are used as a flavour herbal tea and the essential oil was used as one of perfume ingredient. The antioxidative principal-acteoside was isolated from flower extracts of O. fragrans. The chemical stability of crude extract of O. fragrans and acteoside were evaluated by treatment with various pH buffer solutions and different temperature.

Materials and Methods: The flowers of O. fragrans were extracted with ethanol, 50% ethanol and water, respectively. The antioxidative activities were evaluated by DPPH and ABTS scavenging assay. Acteoside was isolated from the 50% ethanol extract of O. fragrans. The thermal stability of crude extract and acteoside aqueous solutions were evaluated at 25 °C, 40 °C, 60 °C, 80 °C, and 90 °C dry bath for 24 h. The pH stability of crude extract and acteoside aqueous solutions were evaluated at pH 4.0, 5.0, 6.0, 7.0, and 8.0 of 0.1M phosphate buffer solution for 7 days.

Results: Acteoside showed potential DPPH and ABTS scavenging activities. The thermal stability results showed both crude extract and acteoside aqueous solution were stable at 25 °C, 40 °C, and 60 °C for 24 h. However, when acteoside aqueous solution was kept at 90 °C, the degradation was significant. The pH stability result showed crude extract and acteoside were more stable at pH 4.0-6.0 buffer solutions and quick degradation at pH more than 7.0.

Conclusions: The different stability properties of O. fragrans crude extract and acteoside in solutions can be used for develop the related herbal tea in the future.


  International Education and Cultural Exchange of Traditional Chinese Medicine Top


Elizabeth R. Qia, Zhong-Zhen Zhaoa, Hu-Biao Chena, Ping Guoa, Jing Liua

aSchool of Chinese Medicine, Hong Kong Baptist University, Kowloon Tong, Kowloon, Hong Kong SAR, China

Correspondence:

Elizabeth R. Qi, E-mail: elizabeth_qi@ hkbu.edu.hk

Materia medica literature can be a significant resource for the modern-day application and research of traditional Chinese medicine. With the commemoration of Li Shizhen's 500th birthday, we used to opportunity to launch cultural projects to educate and disseminate Chinese medicine knowledge. These projects range from traditional media such as books, newspaper, and television as well as exhibitions, museums, and documentaries. The internet has also played an integral role in structuring and promoting international education. In more traditional media, Zhongzhen on Materia Medica is a 20-episode video series, exploring the history, the literature, and the folk customs of traditional medicine. This program was well received in China as well as abroad, having been broadcasted even on television in the United States. Another series, Materia Medica of the World, was a 120-episode television series broadcasted in Hong Kong where a host mediated conversations with experts on traditional Chinese medicine. International education and cultural exchange is needed to further research the knowledge inherited through traditional medicine as well as push modern-day scientific innovations.


  Study on the Human Plasma Protein Binding of Bioactive Components in Sosihotang Soft-Extract Top


Eun-Jeong Choia, Go-Wun Choia, Hee-Woon Janga, Guk-Yeo Leeb, Young-Dal Kwonc, Hea-Young Choa

aCollege of Pharmacy, CHA University, Seongnam-Si, Gyeonggi-Do,bNational Development Institute of Korean Medicine, Jangheung-Gun, Jeollanam-do,cDepartment of Oriental Rehabilitation Medicine, Wonkwang University Gwangju Medical Center, Nam-Gu, Gwangju, Republic of Korea

Correspondence:

Hea-Young Cho, E-mail: hycho@cha.ac.kr

Introduction: Sosihotang is a traditional herbal medicine for fever and inflammatory diseases treatment, and it was developed into soft-extract for patient compliance. The purpose of this study was to evaluate the human plasma protein binding rates of bioactive components of Sosihotang soft-extract such as glycyrrhizin (GL), 18β-glycyrrhetinic acid (GA), baicalin (BG) and baicalein (B) in vitro and in vivo.

Materials and Methods: The ultrafiltration technique was used to determine of plasma protein binding rates of GL, GA, BG and B. For in vitro study, these components were added to blank human plasma to final concentrations of 0.1 and 1 μg/mL. For in vivo study, human plasma samples which were collected at 1 and 8 h after oral administration of Sosihotang soft-extract were used. Each sample was transferred to Centrifree® Ultrafiltration Device and centrifuged at 3000 g for 15 min. Both ultrafiltrates and non-ultrafiltrates were analyzed by UPLC-MS/MS. The human plasma protein binding rates of components were calculated by (1-ultrafiltrates concentration/non-ultrafiltrates concentration) × 100.

Results: In vitro human plasma protein binding rates of these components were calculated about 99%. In vivo human plasma protein binding of GL could not be calculated because it was not detected in samples. Results of BG in clinical samples were similar to in vitro result; however in vivo results of GA and B varied widely,22.64-98.65 and 64.33-95.57%, respectively.

Conclusions: In this study, human plasma protein binding rates of bioactive components of Sosihotang was evaluated in vitro and in vivo. Furthermore, this data can be considered in the evaluation of pharmacokinetics in humans.

Acknowledgements

This work was supported by National Development Institute of Korean Medicine (NIKOM) grant funded by the Korean Ministry of Health and Welfare (MOHW) in 2018.


 Salmonella More Details spp., Staphylococcus aureus, and Pseudomonas aeruginosa in Decoction">  A Real-Time PCR Assay for the Simultaneous Detection of Salmonella spp., Staphylococcus aureus, and Pseudomonas aeruginosa in Decoction Top


Mi-Eun Baeka, Yoon-Jung Choia, Chansook Yoona, Jae-Hee Wona

aNational Development Institute of Korean Medicine, Gyeongsan-si, Gyeongsangbuk-do, Korea

Correspondence:

Jae-Hee Won, E-mail: won10042@nikom.or.kr

Introduction: Salmonella spp., Staphylococcus aureus, and Pseudomonas aeruginosa are dominant human pathogens, so, they are managed by the Korean Pharmacopoeia with 'not detected'. The microbiological culture methods are being used to detect these bacteria in Korean Pharmacopoeia, this method complicated and time-consuming because different bacteria must be used culture medium, differently for each strain and biochemical tests must be performed. So, we tested a rapid and multiplex RT-PCR method to detect these bacteria.

Materials and Methods: Strains were obtained from Korean Collection for Type Cultures. Multiplex Real-Time PCR kit and DNA extraction kit were obtained from KogeneBiotech, and the test was performed according to the manufacturer's instructions. Bio-Rad IQ5 Real-Time PCR was used and tested for optimal the pre-incubation times and the reproducibility.

Results: In order to detect these bacteria in decoction, a pre-incubation time for 12 hrs is required at least, and this method showed repetitive reproducibility. The range of the CT value was 20 to 25, and RFU value determined against the control DNA. The microbiological culture methods take 2 to 7 days, but the RT-PCR Assay was able to confirm the results within 24 hrs.

Conclusions: Our results show that the RT-PCR assay has potential as a method for the detection of Salmonella spp., S. aureus, and P. aeruginosa in decoction, and It is expected to play a significant role in the microbiological quality control of decoction.


  RAM Study on General Standard of Maximum Residue Limits for Pollution-Free Traditional Chinese Medicine Based on Chinese Pharmacopoeia Formula Top


Lu Luoa, Lin-Lin Donga, Meng-Zhi Lia, Yi-Chuan Lianga, Guang-Fei Weia, Yun-Yi Zhanga, Zhong-Zhi Qianb, Shi-Lin Chena

aInstitute of Chinese Materia Medica, China Academy of Chinese Medical Sciences,bChinese Pharmacopoeia Commission, Beijing, China

Correspondence:

Shilin Chen, E-mail: slchen@icmm.ac.cn

The excessive pesticide residues and heavy metals in traditional Chinese medicines seriously endanger human health and the sustainable development of Chinese medicine industry. In order to improve its quality and establish a general standard for maximum residue limits (MRL) of pesticides, the RAND/UCLA Appropriateness Method (RAM) was applied to determine the categories and MRLs. Two questionnaires were drafted for expert panel and appropriateness analysis was carried out with the 9-point Likert scale to determine the general standard for MRLs of non-polluting traditional Chinese medicines and decoction pieces. The results showed that a total of 9 experts from different fields scored the necessity of standard-setting and 206 pesticide residue limits respectively. The return rate of both questionnaires was 100%, so the positive coefficient of experts was high. For the necessity questionnaire, the average value was greater than 7, the coefficient of variation was less than 0.25, and the full score ratio was 0.44-1.00; for the MRLs questionnaire, the average value was greater than 8, the coefficient of variation was less than 0.2. 5, and the full score ratio is 0.33-0.89. By RAM, the appropriateness scores of 206 pesticides were greater than 7, and appropriateness rate was 100%, which signifies that the expert panel has reached consensus, shown in [Figure 1], [Figure 2], [Figure 3]. In summary, based on the RAND/UCLA Appropriate Method, the General Standard for Maximum Residue Limits of Pesticides in Pollution-free Chinese Medicines and Decoction Pieces has reached an expert consensus, which provides a basis for standardizing the use of pesticides in pollution-free Chinese Medicines.
Figure 1: Flow chart of general standard setting of maximum residue levels for pesticides in pollution-free traditional Chinese medicine and decoction pieces

Click here to view
Figure 2: Comparison of pesticide category between pollution-free traditional Chinese medicine and international standards retrieved

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Figure 3: Respective comparison of MRLs of the same pesticide category between Pollution-free Traditional Chinese Medicine and individual International standard

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Keywords: Pesticide residue, pollution-free, RAM, standard


  Prospective Feasibility Clinical Study about Safety of Acupotomy Top


Sang-hoon Yoona,b, Chan-Young Kwonc, Jungtae Leema,d

aChung-Yeon Central Institute,bChung-Yeon Korean Medicine Hospital, Seo-gu, Gwangju,cDepartment of Clinical Korean Medicine, Graduate School, Kyung Hee University, Dongdaemun-gu,dDongshin Korean Medicine Hospital, Yangcheon-gu, Seoul, South Korea

Correspondence:

Jungtae Leem, E-mail: julcho@naver.com

Introduction: Acupotomy is modern-type of acupuncture that inserts a needle with a 0.8-mm-wide blade combined with a 1-mm-diameter needle into the human body. Acupotomy is widely used in several musculoskeletal disease. However, as it is more invasive than conventional type needle, safety issue of acupotomy become important. We conducted a single center prospective observational feasibility study. We will assess appropriateness of the future multi center study protocol.

Materials and Methods: First, literature review was conducted about acupotomy related adverse events (AEs). Based on literature review, Delphi study was conducted to make consensus about definition of each AEs induced by acupotomy. Based on the consensus, standardized case report form was developed. Prospective observational study was conducted to assess follow-up rate, incidence, severity and causality of AEs induced by acupotomy. Descriptive data of each AEs was acquired. Risk factor that associated with AEs will also be evaluated by generalized multinomial (or binomial) mixed logistic regression (Trial Registration: https://cris.nih.go.kr KCT0002666).

Results: A total of 28 participants was participated in the safety study. Total treatment session was 257 with 1,186 treatment points during 4 months. Follow up rate was about 98% which means study protocol was feasible. Total systematic AEs was 8 from 257 treatment sessions (3.11%). And local AEs were 27 from 1186 treatment points (2.28%). There were not severe AEs. Several factors like Number of stimulation and insertion depth were associated with AEs.

Conclusions: According to this preliminary observational study results, the protocol for further prospective safety study is judged to be feasible. We found that acupotomy is relatively safe. However, further multi-center study with more participants needed to support this results. This data will be used fundamental information for further research.

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Acknowledgements

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  Study of Congolese Ethnoveterinary Practices, a Preliminary for the Scientific Evaluation of Medicinal Plants Properties (DR Congo) Top


M. G. Nzuzia,b,c, E. V. Okombea, S. C. Pongomboa, S. Vandenputc, A. Nachtergaelb, S. A. Bakarid, P. Duezb

aFaculty of Veterinary Medicine, University of Lubumbashi,dUniversity of Lubumbashi (UNILU), Faculty of Pharmacy, Lubumbashi, DR Congo,bUnit of Therapeutic Chemistry and Pharmacognosy, University of Mons, Mons,cBibliothèque des Sciences de la vie, University of Liège, Liège, Belgium

Correspondence:

Nzuzi Mavungu Gaël, E-mail: mavungug@unilu.ac.cd

Introduction: Veterinary phytotherapy has not been sufficiently explored as an alternative for the treatment of animal diseases1,2. In the perspective of traditional knowledge valorization, this study investigates the use of herbal medicines to treat animal diseases.

Material and Methods: Ethnobotanical surveys were conducted in three territories (Kasenga, Kipushi and Sakania) of the province of Haut-Katanga (DR Congo) in order to gather information on the use of medicinal plants by farms breeders to treat various diseases. A bibliographic screening of the biological activities reported for these plants was also carried out in Pubmed, Scopus and Google scholar databases.

Results: From the 167 farmers interviewed, 150 (89.9 %) reported the use of traditional medicines among which 89 men and 61 women. The most treated diseases belong to the gastrointestinal sphere, notably helminthiasis. The plants of the Fabaceae family (23 %) are the most used. Roots (26 %) are the most commonly used organs and are administered as aqueous macerates (33 %). There is clearly a need to investigate the anthelminthic use of the most cited plants as only some 35 % of them have been previously studied, but only for antimicrobial, antioxidant and anti-inflammatory properties.

Conclusion: In DR Congo, a large proportion of breeders use traditional medicines to treat animal diseases. Some of the cited plants could be an effective alternative strategy in maintaining animal productivity and are worth of further chemical and biological studies; these will provide indications for promising extract leads that can be developed into standardized medications.

Acknowledgements

The ARES (Academie de Recherche et de l'Enseignement Supérieur) is gratefulyl acknowledged for financial support.


  References Top


  1. Bartha SG, Quave CL, Balogh L, Papp N. Ethnoveterinary practices of Covasna County, Transylvania, Romania. J Ethnobiol Ethnomed 2015;11:35.
  2. Okombe EV, et al. J Ethnopharmacol 2014;119:559-74.



  The Perception of Traditional Medicine by Conventional Health Practitioners in Lubumbashi, Haut-Katanga Province, DR Congo Top


Cedrick Mutombo Shakalengaa,b, Salvius Bakari Amuria, Amandine Nachtergaelb, Pierre Duezb, Kahumba Byangaa

aLaboratory of Pharmacognosy, Faculty of Pharmaceutical Sciences, University de Lubumbashi, Lubumbashi, DR Congo,bUnit of Therapeutic Chemistry and Pharmacognosy, Faculty of Medicine and Pharmacy, University of Mons, Mons, Belgium

Correspondence:

Cedrick Mutombo Shakalenga, E-mail: shakalengam@unilu.ac.cd, cedrick.mutomboshakalenga@student.umons.ac.be

Introduction: The perception of conventional health practitioners on traditional medicine (TM) is an important feature for orienting actions to be carried out for the integration of traditional medicine into the official health system.[1],[2],[3] The present study evaluates the use of traditional medicine by conventional medicine professionals and their opinion on the integration or cohabitation of traditional healers into their structures.

Methodology: A survey was carried out among 196 doctors, 14 pharmacists, 324 nurses and 16 laboratory technicians, between February and July 2018, in 5 health zones of Lubumbashi.

Results: 61.7 % of nurses, 52.6 % of doctors, 78.6 % of pharmacists, and 62.5 % of laboratory technicians had already used traditional medicine. They recognize some diseases for which traditional medicine would be at least as effective as conventional treatments, e.g. epilepsy and arterial hypertension. Those who did not use TM (n=226) report various reasons, notably its irrationality (42 %) and a purported lack of effectiveness (8 %). From the surveyed professionals, 58.9 % of nurses, 57.1 % of doctors, 92.9 % of pharmacists, and 62.5 % of laboratory technicians are willing to collaborate with traditional healers, essentially for complementarity reasons. It is noticeable that 40.8 % of doctors would accept to integrate traditional healers into their care structures.

Conclusion: In Lubumbashi, a majority of conventional medicine practitioners resort to traditional medicine. Most of them recognize the effectiveness of traditional medicine and accept the idea of a cohabitation between traditional and conventional medicines.

Keywords: Collaboration, conventional medicine, DR Congo, Lubumbashi, traditional medicine

Acknowledgements

To the ARES which subsidizes our doctoral training.


  References Top


  1. Krah E, de Kruijf J, Ragno L. Integrating traditional healers into the health care system: Challenges and opportunities in rural Northern Ghana. J Community Health 2018;43:157-63.
  2. ONUSIDA. Collaboration Avec les Guérisseurs Traditionnels Pour la Prévention et la Prise en Charge du VIH en Afrique Subsaharienne: Suggestions à L'intention des Administrateurs de Programme et des Agents de Terrain. Genève, Suisse: ONUSIDA; 2007. p. 58.
  3. Kwaku OJ, Addai-Mensah P, Wiafe F. Traditional and modern medicine: A survey of views on its integration in Ghana. Int J Afr Soc Cult Trad 2015;3:22-36.



  Current Situation of Concentrated Chinese Medicine Granules and its Challenge for Standardization Top


Abraham Y. L. Chana, Hisayoshi Norimotoa

aPuraPharm International Limited, Central, Hong Kong, China

Correspondence:

Dr. Hisayoshi Norimoto, E-mail: hnorimoto@purapharm.com

Concentrated Chinese Medicine Granules (CCMG), also called dispensing granules has been rapidly developed for about 20 years since the Chinese Central government issued “Provisional Regulations on Management of CCMG” in 2001. And the goverment has been stricter in the control of pilot enterprises. Only six companies have obtained this qualification of CCMG manufacturing, including our PuraPharm. As a novel dosage form in characteristics of decocting-free granules in place of Chinese herb slices (YinPian in Chinese), which require a long decoction process, CCMG is not only more convenient to patients, dispensing, transportation and storage in pharmacies or hospitals, but more important is that doctor is able to prescribe different combination of CCMG according to the different constitution of the patients, in turn, to accomplish the goal of “personalized medicine”.

On the other hand, how to establish the standard of CCMG is becoming a very important task because quality is the paramount issue as it can affect the efficacy and/or safety of the products being used. In 2012, the Chinese Pharmacopoeia Commission officially announced building a national standard of CCMG with scientific and comprehensive approaches such as reflecting “intrinsic fingerprint profiles” methods to ensure the efficacy and safety of CCMG products. However, when the pilot enterprises are working on the standard building under the guide of the Pharmacopoeia Commission, they have experienced and met various difficulties and challenges not only in technology including production scale but also in approaches to standardization of quality control.

In this presentation, the speakers want to share the current status of CCMG in China and some experience in building quality control standardization. Meanwhile also want to show personal views on the standard building through investigation of Japanese Kampo standardization, because the Chinese Pharmacopoeia Commission introduced the same concept of “standard decoction” to build the quality control standard.


  Pragmatic Trial for Chinese Medicine: The Conceptual Design and Implementation of a Semi-Individualised Trial for Diabetic Kidney Disease Top


Chan Kam-Waa, Wong Taam Chi-Woon Viviana, Tang Chi-Wai Sydneya

Comparative effectiveness research and pragmatic trial have been advocated across different specialties in the past few decades with the goal to evaluate interventions from a real-world setting. Although pragmatic trial has the advantage of having more flexibility and better clinical application, the balance between such flexibility and scientific rigour including reproducibility and control of bias is controversial and has to be properly addressed.

Clinical application of the data generated by traditional drug trials from population perspective has been questioned consistently. Chinese medicine has a strong emphasis on individualisation, which is coherent to the late call for personalised medicine. Our qualitative study data shows that pragmatic design is generally accepted by clinicians from western medicine and Chinese medicine. The use of real-world data and expert consensus provides a more efficient way of identifying potential regime.

SCHEMATIC, a semi-individualised randomised controlled trial, is one of the earliest registered pragmatic clinical trials of Chinese medicine evaluating clinical protocol as a whole when compared to standard care alone on diabetic kidney disease. The conceptual design including overall structure, choice of intervention, flexibility in protocol, choice of outcome measurement and hierarchy of analysis were based on and optimised by expert consensus, preliminary data from previous service programme and qualitative stakeholder analysis from a patient-centre perspective.

The implementation of SCHEMATIC formed an add-on service component to the existing infrastructure of public clinics as a clinical trial which can be easily translated to routine clinical service. The first half of SCHEMATIC subjects have completed the study and the attrition is well-controlled. The clinical outcome is promising in addressing the demand of both patients and clinicians.


  TCM as Immunomodulators for the Treatment of Rheumatoid Arthritis: Bioactivity and Quality Control Top


Cindy L. H. Yanga, Stanley C. C. Chika, James C. B. Lia, Allan S. Y. Laua, Hisayoshi Norimotob, Godfrey C. F. Chana

aMolecular Chinese Medicine Laboratory, LKS Faculty of Medicine, The University of Hong Kong, Pokfulam,bPuraPharm International Limited, Central, Hong Kong, China

Correspondence:

Prof. Godfrey C. F. Chan, E-mail: gcfchan@hku.hk

Introduction: (Word Count: 279, Maximum=300).

Rheumatoid arthritis (RA) is a chronic, systemic autoimmune inflammatory disorder with destruction of articular cartilage and erosion of bone in the joints, leading to loss of function and mobility of the patients. Existing therapeutics are suboptimal, expensive and associated with serious side effects.

Cimicifuga racemose has strong reputations in treating RA by native American. In this study, we hypothesize that its Chinese counterparts (Shengma) contains similar novel anti-inflammatory compounds. These novel compounds can be used as chemical markers for the quality control of Shengma.

Materials and Methods: We isolated and identified three anti-inflammatory compounds from Shengma using Chromatography and Nuclear Magnetic Resonance. Human primary blood macrophages (PBMac) were used to investigate the anti-inflammatory effect of newly identified compounds by lipopolysaccharides (LPS)/ lipopeptide Pam3CSK4-induced cytokine production assays. Cytokine expression was measured by quantitative RT-PCR and ELISA. Collagen antibody induced arthritis (CAIA) Balb/c mice model was used to test the in vivo effect of Shengma.

Results: Several compounds were identified including Cim A, PSM-6 and PSM-7. Cim A could suppress the LPS-induced TNF-α production in PBMac, whereas PSM-6 and PSM-7 suppressed Pam3CSK4-induced TNF-a and IL-6 production in PBMac dose-dependently. The anti-inflammatory activity of Cim A was due to its modulation of signaling mitogen activated protein kinase and transcription factor nuclear factor-kappaB activities. Mice treated with Shengma granules showed lower disease scores and lower mRNA levels of the inflammatory cytokines and chemokines in the paw joints when compared to the control mice. Cim A, PSM-6 and PSM-7 could be used as the chemical markers for the quality control of Shengma.

Conclusion: Our findings have clinical implications for the potential use of Shengma or its active compounds as alternative therapeutics for RA.

Acknowledgement

This project was supported by PuraPharm International Limited.


  Is it Safe to Use Estrogenic Herbs in Breast Cancer Patients? Top


Clara Bik-San Laua

aInstitute of Chinese Medicine and State Key Laboratory of Research on Bioactivities and Clinical Applications of Medicinal Plants, The Chinese University of Hong Kong, Hong Kong, China

Correspondence:

Clara Bik-San Lau,

Estrogenic compound-containing herbs, including several commonly used Chinese herbal medicines, are believed to possess beneficial effects for women's health. However, contrastive views have been suggesting the stimulating activities of estrogenic herbs in breast cancer progression, which raised alertness of breast cancer patients, survivors as well as Chinese medicine practitioners, and resulted in hindering the use of estrogenic herbs, particularly Angelica sinensis in breast cancer patients over the decades. In view of the impossibility to conduct clinical trials for evaluating safety of estrogenic herbs in breast cancer patients, we aimed to systematically investigate the potential stimulating effects of estrogenic herbs on breast cancer growth using both preclinical cell-based and tumor-bearing mouse models.

In this study, eleven well-known estrogenic herbs, including Angelica sinensis (AS), Astragalus membranaceus, Panax ginseng, etc. were firstly evaluated in homogeneous breast cancer cell lines with different molecular subtypes for their proliferative responses. Those with potent stimulatory activities were further verified in primary breast cancer cells isolated from patients, and the systemic responses towards these herbs were determined in mice bearing human breast xenografts or mouse syngeneic tumors. Our results showed that a few tested estrogenic herbs increased the viability of breast cancer cells lines in different extents, such as AS and Cistanche deserticola (CD). Despite transient stimulatory activities of AS on breast tumor growth was observed in mice, the final tumor weights were not significantly altered after 4 weeks of AS or CD treatments.

In conclusion, a novel approach of applying breast cancer cell lines, xenograft and syngeneic tumors models, as well as primary breast cancer cells from patients' tumors in herbal medicines safety evaluation was established. Our findings also provided comprehensive preclinical evidences to disprove the paradoxical perception of Angelica sinensis in promoting breast cancer growth, which may affect future clinical practice in breast cancer patients.


  References Top


  1. Yue GG, Wong LS, Leung HW, Gao S, Tsang JY, Lin ZX, et al. Evaluation of the safety profiles of estrogenic Chinese herbal medicines in breast cancer. Phytomedicine 2019;56:103-17.
  2. Yue GG, et al. Frontiers Pharmacol 2019.



  How do we Ensure Supply of Qood Quality Plants for TCM? Top


Monique S. J. Simmondsa

aRoyal Botanic Gardens, Kew, United Kingdom

In a world facing challenges associated with the impact of climate change, that includes extreme weather events, increased pests and failure of natural pollinators as well as changes in land use to feed increasing populations, how do we ensure we have a sustainable supply of plants for use in traditional Chinese medicine (TCM)? As the interest in plants used in TCM increases this challenge increases. Meeting this challenge underpins the work of many scientists attending this conference. China should be congratulated in the huge efforts it is making to conserve its genetic resources as it is the only country in the world that has seen a significant increase in the expansion of botanic gardens that supports this conservation work. However, is this enough? This talk will provide a western perspective of these challenges and how the Good Practice in traditional Chinese medicine Research Association (GP-TCM RA) could have an impact in helping to map a route to support the sustainable supply as well as the collation of data that helps specify the criteria that defines what a “quality” plant is. The importance of understanding “quality” is a key to the selection of plants and the products that are made from them.

I would like to dedicate this talk to Professor Peter Hylands of King's College London. Peter was a founder of the EU-China project that led to the foundation of this society. We would not all be here at this event if it had not been for the support that Peter gave to the founding team led by Qihe Xu.


  Cinnamomum cassia Promotes Apoptosis of Lung Cancer Cells through PDK1 Inhibition Top


Eun-Ji Leea,b,c, Tae-Wook Chunga,b, Jung-Hee Leeb, Bo-Sung Kima,b,c, Eun-Yeong Kima,b,c, Syng-Ook Leed, Ki-Tae Haa,b,c

aDepartment of Korean Medical Science, School of Korean Medicine, Pusan National University,bHealthy Aging Korean Medical Research Center, Pusan National University,cGraduate Training Program of Korean Medicine for Healthy-aging, Pusan National University, Yangsan, Gyeongnam,dDepartment of Food Science and Technology, Keimyung University, Dalgubeoldae-ro, Daegu, Republic of Korea

These authors contributed equally in this study.

Correspondence:

Ki-Tae Ha, E-mail: hagis@pusan.ac.kr

Introduction: Cinnamomum cassia Blume has been widely reported as the anti-tumor agent. However, the precise mechanism underlying its pro-apoptotic action is still not clear. Restraining aerobic glycolysis through suppression of pyruvate dehydrogenase kinase 1 (PDK1) is a promising strategy for cancer inhibition. This study was performed to investigate the anti-tumor action of C. cassia is mediated by PDK1 inhibition.

Materials and Methods: The inhibition of water-extracted branch of C. cassia (WBCC) on the activity of PDK1 using both in vitro and cell-based kinase assay were examined in several lung cancer cells. The viabilities of cells treated with WBCC were evaluated by cell counting and MTT assay. Pro-apoptotic action of WBCC was examined by PI-stained DNA content analysis and Western blot. Mitochondrial membrane potential was measured TMRM staining. The production of intracellular and mitochondrial ROS were detected by carboxy-H2DCFDA and MitoSOX, respectively. The major components of WBCC were estimated by HPLC analysis.

Results: WBCC reduced viabilities of several lung cancer cells with minimal cytotoxicity on normal bronchial epithelial cells. WBCC decreased lactate production through inhibiting activity of PDK1 but not affect lactate dehydrogenase A. In consequence of PDK1 inhibition, WBCC increased ROS production, which damage mitochondria membrane stability. In addition, WBCC induced ROS- and mitochondria-dependent apoptotic cell death. Among the components of WBCC, cinnamic acid was founded as a major inhibitor on PDK1 activity.

Conclusions: This is first report that WBCC induces apoptosis of lung cancer cells through inhibiting PDK1 activity. Our findings suggest that WBCC and cinnamic acid can be potential candidates for developing novel anti-cancer drugs through glycolysis metabolism.


  Paeoniflorin Increases the Adhesion of Trophoblast to the Endometrium by Upregulating the Expression of Integrin αVβ3 and αVβ5 in LIF-Dependent Manner Top


Hye-Rin Parka,b, Hee-Jung Choia, Mi-Ju Parka, Ki-Tae Haa,b

aKorean Medical Research Center for Healthy Aging,bDepartment of Korean Medical science, School of Korean Medicine, Pusan National University, Yangsan, Gyeongsangnam-do, Republic of Korea

These authors contributed equally in this study.

Correspondence:

Ki-Tea Ha, E-mail: hagis@pusan.ac.kr

Introduction: Uterine receptivity, which is determined by diverse biological factors such as adhesion molecules, cytokines, growth factors, and receptors, is one of the important factors for successful implantation. Our previous study has shown that water extract of Paeonia lactiflora enhanced embryo implantation in vitro and in vivo via induction of the leukemia inhibitory factor (LIF)-dependent expression of Integrin αVβ3 and αVβ5.

Methods and Results: We performed high-performance liquid chromatography (HPLC) analysis to investigate which one is the major component. Next, we tested these five compounds to confirm whether these single compounds enhance the adhesion of human trophectoderm-derived JAr cells to the endometrial Ishikawa cells. In addition, we checked the expression of adhesion molecules in mRNA and protein levels and performed in vivo study using the implantation failure model mice by treating RU-486.

Paeoniflorin, the most abundant molecule among tested five major compounds of P. lactiflora, showed enhancing effect on cellular interaction between JAr and Ishikawa cells similar to Paeonia lactiflora. Paeoniflorin increased the expression of Integrin αVβ3 and αVβ5 and in the case of integrin αVβ5, the expression level was regulated in LIF-dependent manner. Furthermore, in vivo study showed that paeoniflorin significantly improved the number of implantation embryos.

Conclusion: Thus, our data demonstrates that paeoniflorin, a major compound of P. lactiflora, is the most crucial compound of P. lactiflora in enhancing endometrial receptivity.

Acknowledgements

This study was supported by a National Research Foundation of Korea (RNRF) grant funded by the Ministry of Science, ICT & Future Planning (MISP), Korea Government.


  Extracts of Achyranthis Radix and Acanthopanacis Cortex Promote Hair Growth in Human Hair Follicle Dermal Papilla Cells via Transition of Hair Cycle Top


Yoo Jin Kima, Mi Jin Jeonga, Mi Ryeo Kima

aDepartment of Herbal Pharmacology, College of Korean Medicine, Daegu Haany University, Daegu, Korea

Correspondence:

Mi Ryeo Kim, E-mail: mrkim@dhu.ac.kr

Introduction: According to the number of people who are interested in appearance, hair has become one of the important aspects of beauty. There are also many studies about prevention and treatment of alopecia. Finasteride and minoxidil have been known for drugs for alopecia treatment. However, these agents have side effects such as irritation and erythema when used for a long time. Thus, we need more study on herbal medicine for hair growth. In the previous study, herbal mixture (JHU) included in Rehmanniae Radix Preparata (RRP), Achyranthis Radix (AR), and Acanthopanacis Cortex (AC) promoted hair growth in vitro and in vivo. Thus, this study investigated the effects of those three components (RRP, AR, and AC) on hair growth in human hair follicle dermal papilla cells (HDPC).

Materials and Methods: The proliferation was determined by MTT assay in HDPC. We evaluated cell cycle of HDPC treated RRP, AR, and AC using flow cytometry. Hair growth-related gene expressions monitored by western blot.

Results: Extracts of AR and AC, components of mixture, were increased cell proliferation and transition of hair cycle compared with RRP, respectively. AR and AC also were more increased protein expression than RRP, respectively.

Conclusions: AR is also known not only to prevent grey hair, but also hair loss through promoting blood circulation and haematopoiesis. AC helps blood circulation because of having warm nature and spicy taste. Thus, AR and AC promoted hair growth and they could be used as a natural material for treatment and prevention of hair loss.

Acknowledgements

This work was carried out with the support of “Cooperative Research Program for Agriculture Science & Technology Development (Project No. 18-113-001)” Rural Development Administration, Republic of Korea.


  The Effects of Plantago asiatica L. in High-Fat Diet-Induced Obese C57BL/6 Mice via Controlling Lipid Metabolism Top


Mijin Jeonga, Yoojin Kima, Sungok Kima, Yeji Leea, Minyoung Yoona, Mi Ryeo Kima

aDepartment of Herbal Pharmacology, College of Korean Medicine, Daegu Haany University, Daegu, Republic of Korea

Correspondence:

Mi Ryeo Kim, E-mail: mrkim@dhu.ac.kr

Introduction: Plantago asiatica L. has been used for treatment of inflammation, bronchitis, cholesterolemia, chronic constipation, and diarrhea in Korea. In the present study, the anti-obesity effects of Plantago asiatica L. water extract (PAE) were investigated.

Materials and Methods: After a week of adaptation, mice were fed either a normal diet (ND) or a high-fat diet (HFD). PAE was administered orally for 6 weeks to some of the mice fed the HFD. Body weight, food intake, and white adipose tissue weight (WAT) were measured. An enzyme reaction experiment was used for biochemical plasma analysis. Protein expressions related to adipogenesis or lipid metabolism were evaluated in liver by western blot. In addition, UPLC analysis was used for determination of candidates of active ingredients in PAE.

Results: Body weight, body weight gain, food efficiency rate (FER), and total WAT weight significantly decreased in the PAE groups compared with the HFD group. The lipid contents in the plasma significantly decreased in the PAE groups. The protein expression levels of hepatic p-AMPK, p-ACC and CPT-1 increased but those of PPARγ, SREBP-1c and HMG-CoA decreased in the PAE groups compared to the HFD group. Candidates of active ingredients in Plantago asiatica L. were shown plantamajoside and acteoside through UPLC.

Conclusions: Our results indicated that Plantago asiatica L. is effective to prevent and to improve obesity, so it might be a potential and safe material for anti-obesity agent or functional supplements. We suppose that plantamajoside and acteoside may be candidates of active ingredients in PAE.

Acknowledgements

This research was supported by Basic Science Research Program through the National Research Foundation of Korea(NRF) funded by the Ministry of Science, ICT & Future Planning (2018R1A2B6009036).


  Six-Sialyllactose Ameliorates Dihydrotestosterone -Induced Benign Prostatic Hyperplasia through Suppressing VEGF-Mediated Angiogenesis Top


Eun-Yeong Kima,b, Bo-Ram Jinc, Tae-Wook Chungb, Sung-Jin Baeb, Hyerin Parka,b, Dongryeol Ryua,b, Ling Jina,b, Hyo-Jin Anc, Ki-Tae Haa,b

aDepartment of Korean Medical Science, School of Korean Medicine,bKorean Medical Research Center for Healthy Aging, Pusan National University, Yangsan, Gyeongsangnam-do,cDepartment of Pharmacology, College of Korean Medicine, Sangji University, Gangwon-do, Republic of Korea

Correspondence:

Hyo-Jin An, E-mail: hjan@sj.ac.kr

Ki-Tae Ha, E-mail: hagis@pusan.ac.kr

Benign prostatic hyperplasia (BPH), a common disease for elder man, is accompanied by nonmalignant growth of prostate tissues, subsequently causes hypoxia and angiogenesis. Although VEGF-related angiogenesis is one of therapeutic target of prostate cancer, there is no previous study targeting angiogenesis to treat BPH. Dihydrotestosterone (DHT)-induced expressions of vascular endothelial growth factor (VEGF) in prostate epithelial RWPE-1 cells and human umbilical vascular endothelial cells (HUVECs). The conditioned media (CM) from DHT-treated RWPE-1 cells were transferred to HUVECs. Then, 6SL inhibited the proliferation, VEGFR-2 activation, and tube formation of HUVECs which were transferred with CM from DHT-treated RWPE-1 cells. In rat BPH model, 6SL reduced prostate weight, size, and thickness of the prostate tissue. The formation of vessels in prostatic tissues were also reduced by 6SL treatment. We found that 6SL has an ameliorative effect on both of in vitro and in vivo BPH model via inhibition of VEGFR-2 activation and subsequent angiogenesis. These results suggest that 6SL might be a candidate for development of novel BPH drugs.


  Caesalpinia sappan Induces Apoptosis of Ectopic Endometrial Cells by Decreasing Expression of Pyruvate Dehydrogenase Kinase 1 Top


Bo-Sung Kima,b, Tae-Wook Chungb, Hee-Jung Choib, Mi-Ju Parkb, Eun-Yeong Kima,b, Eun-ji Leea,b, Jung-Hee Leeb, Syng-Ook Leec, Jung-Hye Choid, Ki-Tae Haa,b

aDepartment of Korean Medical Science and Graduate Training Program of Korean Medicine for Healthy-aging, School of Korean Medicine,bHealthy Aging Korean Medical Research Center, Pusan National University, Yangsan, Gyeongnam,cDepartment of Food Science and Technology, Keimyung University, Daegu,dDepartment of Life and Nanopharmaceutical Sciences and Department of Oriental Pharmacy Kyung Hee University, Seoul, Republic of Korea

Correspondence:

Ki-Tae Ha,

Endometriosis is a common gynecological disease defined as growth of endometrial tissues outside uterus. Precise mechanism responsible for the progression of endometriosis are still not clear. Because the survival of endometrial cells refluxed into the abdominal cavity develops into endometriosis, inducing endometrial apoptosis is an important strategy for treating endometriosis. Heartwood of Caesalpinia sappan L. (belong to Leguminosae) have been a medicinal herb used for improving blood circulation, accelerating hemostasis, removing of extravasated blood, relieving pain, and reducing swelling. Especially, heartwood of C. sappan was used for treating gynecological symptoms including algomenorrhea and amenorrhea. The endometriotic cells (12Z) grew faster compared with normal endometrial stromal cells (THESC). The expression of TGF-β, pyruvate dehydrogenase kinase (PDK) 1 and 3, and lactate production were higher in 12Z cells. In addition, the 12Z cells were more sensitive to water extracted heartwood of C. sappan (CS)-induced cytotoxicity compared with THESCs. CS inhibited the lactate production and phosphorylation of pyruvate dehydrogenase A through reducing the expression of PDK1. CS also increased the reactive oxygen species (ROS) through activating oxidative phosphorylation and decreased mitochondrial membrane potential. Consequently, CS causes the apoptotic cell death in 12Z cells through ROS and mitochondria dependent manner. In conclusion, CS can be a candidate for development of novel drug treating endometriosis through inhibiting aerobic glycolysis and inducing ROS-mitochondria mediated apoptotic cell death


  Catechin of Inhibition Aerobic Glycolysis and Suppresses Growth of the Drug Resistant Gastric Cancer Cells Top


Jung Ho Hana, Tae-Wook Chungb, Jung-Hee Leeb, Eun-Yeong Kimb, Syng-Ook Leec, Dongryeol Ryua,b, Ki-Tae Haa,b

aDepartment of Korean Medical Science and Graduate Training Program of Korean Medicine for Healthy-aging, School of Korean Medicine,bHealthy Aging Korean Medical Research Center, Pusan National University, Yangsan, Gyeongnam,cDepartment of Food Science and Technology, Keimyung University, Daegu, Republic of Korea

Correspondence:

Ki-Tae Ha, E-mail: hagis@pusan.ac.kr

Normal cells produce adenosine triphosphate (ATP) as cellular energy source through tricarboxylic acid (TCA) cycle. However, cancer cells favor aerobic glycolysis instead of TCA cycle. The phenomena, called as Warburg's effect, is induced by activation of lactate dehydrogenase A (LDHA) or pyruvate dehydrogenase kinase (PDHK). Thus, inhibition of these enzymes was reported as a promising strategy for suppressing tumor growth and resistance. Ongoing screening of herbal medicines and its ingredient components, we found that the roots of Sanguisorba officinalis L. was effective on inhibition of aerobic glycolysis. S. officinalis is an herbal medicine traditionally used for hemostasis, antipyretic, antipyrotic, antidiarrheic. In this study, the water extracted roots of S. officinalis (SO) reduced the activities of LDHA and expression of PDHK4 in human gastric cancer SNU620 cells. In SNU620/5FU, a resistant cell line to 5-fluorouracil (5-FU), co-treatment of SO with 5-FU increased the cytotoxicity. To identify the precise molecular mechanism underlying overcoming 5-FU resistance by SO treatment, we confirmed the expressional levels of several proteins related in 5-FU resistance. Among these proteins, the level of p53, which was suppressed in SNU620/5FU cells, was restored by SO treatment. Among the major compounds of SO including catechin, gallic acid, gallotanin, hamamelitannin, and ellagic acid, we found that catechin is a major active molecules for inhibiting activity of LDHA and phosphorylation of pyruvate dehydrogenase A (PDHA). From these results we suggest SO as a candidate for developing novel adjuvant drug reducing resistance to 5-FU through restricting aerobic glycolysis.


  Huzhangoside a Inhibits Pyruvate Dehydrogenase Kinase Activity and Tumor Growth Top


Choong-Hwan Kwaka, Jung-Hee Leea, Eun-Yeong Kima,b, Chang Woo Hanc, Keuk-Jun Kimd, Hanna Leee, MyoungLae Choe, Se Bok Jangc, Cheorl-Ho Kimf, Tae-Wook Chunga, Ki-Tae Haa,b

aKorean Medical Research Center for Healthy Aging,bDepartment of Korean Medical Science, School of Korean Medicine, Pusan National University, Yangsan, Gyeongsangnam-do,cDepartment of Molecular Biology, College of Natural Sciences, Pusan National University, Geumjeong-gu, Busan 46241, Republic of KoreadDepartment of Clinical Pathology, TaeKyeung University, Gyeongsan,eNational Development Institute of Korean Medicine, Gyeongsan, Gyeongsanabuk-do,fDepartment of Biological Science, Sungkyunkwan University, Suwon, Kyunggi-do, Republic of Korea

Correspondence:

Tae-Wook Chung, E-mail: twchung@pusan.ac.kr

Ki-Tae Ha, E-mail: hagis@pusan.ac.kr

Aerobic glycolysis is one of the important metabolic characteristics of many malignant tumors. PyruvateY dehydrogenase kinase (PDHK) plays a key role in aerobic glycolysis by phosphorylating the E1α subunit of pyruvate dehydrogenase (PDH). Hence, PDHK has been recognized as a molecular target for cancer treatment. Here, we report that huzhangoside A (Hu.A), a triterpenoid glycoside compound isolated from several plants of the Anemone genus, acts as a novel PDHK inhibitor. Hu.A was found to decrease the cell viability of MDA-MB-231, Hep3B, HT-29, DLD-1, and LLC cell lines. The activity of PDHK1 was decreased by Hu.A in both in vitro assays and in vivo assays in DLD-1 cells. Hu.A significantly increased the oxygen consumption and decreased the secretory lactate levels in DLD-1 cells. In addition, Hu.A interacted with the ATP-binding pocket of PDHK1 without affecting the interaction of PDHK1 and pyruvate dehydrogenase complex (PDC) subunits. Furthermore, Hu.A significantly induced mitochondrial reactive oxygen species (ROS) and depolarized the mitochondrial membrane potential in DLD-1 cells. Consistently, when Hu.A was intraperitoneally injected into LLC allograft mice, the tumor growth was significantly decreased. In conclusion, Hu.A suppressed the growth of tumors in both in vitro and in vivo models via inhibition of PDHK activity.




    Figures

  [Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5], [Figure 6], [Figure 7]



 

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