Cinnamaldehyde inhibits the NLRP3 inflammasome by preserving mitochondrial integrity and augmenting autophagy in Shigella sonnei-infected macrophages.

Shigella sonnei Autophagy Cinnamaldehyde NLRP3 inflammasome Pyroptosis

Journal

Journal of inflammation (London, England)
ISSN: 1476-9255
Titre abrégé: J Inflamm (Lond)
Pays: England
ID NLM: 101232234

Informations de publication

Date de publication:
05 Jun 2024
Historique:
received: 16 07 2023
accepted: 22 05 2024
medline: 6 6 2024
pubmed: 6 6 2024
entrez: 5 6 2024
Statut: epublish

Résumé

Worldwide, more than 125 million people are infected with Shigella each year and develop shigellosis. In our previous study, we provided evidence that Shigella sonnei infection triggers activation of the NACHT, LRR, and PYD domain-containing protein 3 (NLRP3) inflammasome in macrophages. NLRP3 inflammasome is responsible for regulating the release of the proinflammatory cytokines interleukin (IL)-1β and IL-18 through the protease caspase-1. Researchers and biotech companies have shown great interest in developing inhibitors of the NLRP3 inflammasome, recognizing it as a promising therapeutic target for several diseases. The leaves of Cinnamomum osmophloeum kaneh, an indigenous tree species in Taiwan, are rich in cinnamaldehyde (CA), a compound present in significant amounts. Our aim is to investigate how CA affects the activation of the NLRP3 inflammasome in S. sonnei-infected macrophages. Macrophages were infected with S. sonnei, with or without CA. ELISA and Western blotting were employed to detect protein expression or phosphorylation levels. Flow cytometry was utilized to assess H CA inhibited the NLRP3 inflammasome in S. sonnei-infected macrophages by suppressing caspase-1 activation and reducing IL-1β and IL-18 expression. CA also inhibited pyroptosis by decreasing caspase-11 and Gasdermin D activation. Mechanistically, CA reduced lysosomal damage and enhanced autophagy, while leaving mitochondrial damage, mitogen-activated protein kinase phosphorylation, and NF-κB activation unaffected. Furthermore, CA significantly boosted phagocytosis and the bactericidal activity of macrophages against S. sonnei, while reducing secretion of IL-6 and tumour necrosis factor following infection. CA shows promise as a nutraceutical for mitigating S. sonnei infection by diminishing inflammation and enhancing phagocytosis and the bactericidal activity of macrophages against S. sonnei.

Sections du résumé

BACKGROUND BACKGROUND
Worldwide, more than 125 million people are infected with Shigella each year and develop shigellosis. In our previous study, we provided evidence that Shigella sonnei infection triggers activation of the NACHT, LRR, and PYD domain-containing protein 3 (NLRP3) inflammasome in macrophages. NLRP3 inflammasome is responsible for regulating the release of the proinflammatory cytokines interleukin (IL)-1β and IL-18 through the protease caspase-1. Researchers and biotech companies have shown great interest in developing inhibitors of the NLRP3 inflammasome, recognizing it as a promising therapeutic target for several diseases. The leaves of Cinnamomum osmophloeum kaneh, an indigenous tree species in Taiwan, are rich in cinnamaldehyde (CA), a compound present in significant amounts. Our aim is to investigate how CA affects the activation of the NLRP3 inflammasome in S. sonnei-infected macrophages.
METHODS METHODS
Macrophages were infected with S. sonnei, with or without CA. ELISA and Western blotting were employed to detect protein expression or phosphorylation levels. Flow cytometry was utilized to assess H
RESULTS RESULTS
CA inhibited the NLRP3 inflammasome in S. sonnei-infected macrophages by suppressing caspase-1 activation and reducing IL-1β and IL-18 expression. CA also inhibited pyroptosis by decreasing caspase-11 and Gasdermin D activation. Mechanistically, CA reduced lysosomal damage and enhanced autophagy, while leaving mitochondrial damage, mitogen-activated protein kinase phosphorylation, and NF-κB activation unaffected. Furthermore, CA significantly boosted phagocytosis and the bactericidal activity of macrophages against S. sonnei, while reducing secretion of IL-6 and tumour necrosis factor following infection.
CONCLUSION CONCLUSIONS
CA shows promise as a nutraceutical for mitigating S. sonnei infection by diminishing inflammation and enhancing phagocytosis and the bactericidal activity of macrophages against S. sonnei.

Identifiants

pubmed: 38840105
doi: 10.1186/s12950-024-00395-w
pii: 10.1186/s12950-024-00395-w
doi:

Types de publication

Journal Article

Langues

eng

Pagination

18

Subventions

Organisme : National Science and Technology Council, Taiwan
ID : MOST 111-2628-B-197-001-MY3
Organisme : National Science and Technology Council, Taiwan
ID : MOST 111-2320-B-532-002

Informations de copyright

© 2024. The Author(s).

Références

Zhang Y, Yang W, Li W, Zhao Y. NLRP3 inflammasome: checkpoint connecting innate and adaptive immunity in Autoimmune diseases. Front Immunol. 2021;12:732933.
pubmed: 34707607 pmcid: 8542789 doi: 10.3389/fimmu.2021.732933
Wang L, Hauenstein AV. The NLRP3 inflammasome: mechanism of action, role in disease and therapies. Mol Aspects Med. 2020;76:100889.
pubmed: 32859386 doi: 10.1016/j.mam.2020.100889
Guo H, Callaway JB, Ting JP. Inflammasomes: mechanism of action, role in disease, and therapeutics. Nat Med. 2015;21(7):677–87.
pubmed: 26121197 pmcid: 4519035 doi: 10.1038/nm.3893
Liao PC, Chao LK, Chou JC, Dong WC, Lin CN, Lin CY, Chen A, Ka SM, Ho CL, Hua KF. Lipopolysaccharide/adenosine triphosphate-mediated signal transduction in the regulation of NLRP3 protein expression and caspase-1-mediated interleukin-1β secretion. Inflamm Res. 2013;62(1):89–96.
pubmed: 22986467 doi: 10.1007/s00011-012-0555-2
Schroder K, Tschopp J. The inflammasomes. Cell. 2010;140(6):821–32.
pubmed: 20303873 doi: 10.1016/j.cell.2010.01.040
He Y, Hara H, Núñez G. Mechanism and regulation of NLRP3 inflammasome activation. Trends Biochem Sci. 2016;41(12):1012–21.
pubmed: 27669650 pmcid: 5123939 doi: 10.1016/j.tibs.2016.09.002
Paik S, Kim JK, Silwal P, Sasakawa C, Jo EK. An update on the regulatory mechanisms of NLRP3 inflammasome activation. Cell Mol Immunol. 2021;18(5):1141–60.
pubmed: 33850310 pmcid: 8093260 doi: 10.1038/s41423-021-00670-3
Takahama M, Akira S, Saitoh T. Autophagy limits activation of the inflammasomes. Immunol Rev. 2018;281(1):62–73.
pubmed: 29248000 doi: 10.1111/imr.12613
Baker S, The HC. Recent insights into Shigella. Curr Opin Infect Dis. 2018;31(5):449–54.
pubmed: 30048255 pmcid: 6143181 doi: 10.1097/QCO.0000000000000475
Daskalakis DC, Blaser MJ. Another perfect storm: Shigella, men who have sex with men, and HIV. Clin Infect Dis. 2007;44(3):335–7.
pubmed: 17205437 doi: 10.1086/510591
Hoffmann C, Sahly H, Jessen A, Ingiliz P, Stellbrink HJ, Neifer S, Schewe K, Dupke S, Baumgarten A, Kuschel A, Krznaric I. High rates of quinolone-resistant strains of Shigella sonnei in HIV-infected MSM. Infection. 2013;41(5):999–1003.
pubmed: 23852945 doi: 10.1007/s15010-013-0501-4
Chiou CS, Izumiya H, Kawamura M, Liao YS, Su YS, Wu HH, Chen WC, Lo YC. The worldwide spread of ciprofloxacin-resistant Shigella sonnei among HIV-infected men who have sex with men, Taiwan. Clin Microbiol Infect. 2016;22(4):383. e11-6.
doi: 10.1016/j.cmi.2015.12.021
Li LH, Chen TL, Chiu HW, Hsu CH, Wang CC, Tai TT, Ju TC, Chen FH, Chernikov OV, Tsai WC, Hua KF. Critical role for the NLRP3 inflammasome in mediating IL-1β production in Shigella sonnei-infected macrophages. Front Immunol. 2020;11:1115.
pubmed: 32582195 pmcid: 7283925 doi: 10.3389/fimmu.2020.01115
Chang ST, Chen PF, Chang SC. Antibacterial activity of leaf essential oils and their constituents from Cinnamomum osmophloeum. J Ethnopharmacol. 2001;77:123–7.
pubmed: 11483389 doi: 10.1016/S0378-8741(01)00273-2
Chang ST, Cheng SS. Antitermitic activity of leaf essential oils and components from Cinnamomum osmophleum. J Agric Food Chem. 2002;50:1389–92.
pubmed: 11879008 doi: 10.1021/jf010944n
Cheng SS, Liu JY, Tsai KH, Chen WJ, Chang ST. Chemical composition and mosquito larvicidal activity of essential oils from leaves of different Cinnamomum osmophloeum provenances. J Agric Food Chem. 2004;52(14):4395–400.
pubmed: 15237942 doi: 10.1021/jf0497152
Wang SY, Yang CW, Liao JW, Zhen WW, Chu FH, Chang ST. Essential oil from leaves of Cinnamomum osmophloeum acts as a xanthine oxidase inhibitor and reduces the serum uric acid levels in oxonate-induced mice. Phytomedicine. 2008;15(11):940–5.
pubmed: 18693097 doi: 10.1016/j.phymed.2008.06.002
Chao LK, Hua KF, Hsu HY, Cheng SS, Lin IF, Chen CJ, Chen ST, Chang ST. Cinnamaldehyde inhibits pro-inflammatory cytokines secretion from monocytes/macrophages through suppression of intracellular signaling. Food Chem Toxicol. 2008;46(1):220–31.
pubmed: 17868967 doi: 10.1016/j.fct.2007.07.016
Liao BC, Hsieh CW, Liu YC, Tzeng TT, Sun YW, Wung BS. Cinnamaldehyde inhibits the tumor necrosis factor-alpha-induced expression of cell adhesion molecules in endothelial cells by suppressing NF-kappaB activation: effects upon IkappaB and Nrf2. Toxicol Appl Pharmacol. 2008;229(2):161–71.
pubmed: 18304597 doi: 10.1016/j.taap.2008.01.021
Guo JY, Huo HR, Zhao BS, Liu HB, Li LF, Ma YY, Guo SY, Jiang TL. Cinnamaldehyde reduces IL-1beta-induced cyclooxygenase-2 activity in rat cerebral microvascular endothelial cells. Eur J Pharmacol. 2006;537(1–3):174–80.
pubmed: 16624280 doi: 10.1016/j.ejphar.2006.03.002
Ho SC, Chang YH, Chang KS. Structural moieties required for cinnamaldehyde-related compounds to inhibit canonical IL-1β secretion. Molecules. 2018;23(12).
Xu F, Wang F, Wen T, Sang W, Wang D, Zeng N. Inhibition of NLRP3 inflammasome: a new protective mechanism of cinnamaldehyde in endotoxin poisoning of mice. Immunopharmacol Immunotoxicol. 2017;39(5):296–304.
pubmed: 28762847 doi: 10.1080/08923973.2017.1355377
Kang LL, Zhang DM, Ma CH, Zhang JH, Jia KK, Liu JH, Wang R, Kong LD. Cinnamaldehyde and allopurinol reduce fructose-induced cardiac inflammation and fibrosis by attenuating CD36-mediated TLR4/6-IRAK4/1 signaling to suppress NLRP3 inflammasome activation. Sci Rep. 2016;6:27460.
pubmed: 27270216 pmcid: 4897702 doi: 10.1038/srep27460
Li LH, Lin JS, Chiu HW, Lin WY, Ju TC, Chen FH, Chernikov OV, Liu ML, Chang JC, Hsu CH, Chen A, Ka SM, Gao HW, Hua KF. Mechanistic insight into the activation of the NLRP3 inflammasome by Neisseria gonorrhoeae in macrophages. Front Immunol. 2019;10:1815.
pubmed: 31417575 pmcid: 6685137 doi: 10.3389/fimmu.2019.01815
Hsieh CY, Li LH, Lam Y, Fang Z, Gan CH, Rao YK, Chiu HW, Wong WT, Ju TC, Chen FH, Chernikov OV, Liu ML, Hsu CH, Hua KF. Synthetic 4-Hydroxy auxarconjugatin B, a Novel Autophagy Inducer, attenuates gouty inflammation by inhibiting the NLRP3 inflammasome. Cells. 2020;9(2):279.
pubmed: 31979265 pmcid: 7072356 doi: 10.3390/cells9020279
Kayagaki N, Stowe IB, Lee BL, O’Rourke K, Anderson K, Warming S, Cuellar T, Haley B, Roose-Girma M, Phung QT, Liu PS, Lill JR, Li H, Wu J, Kummerfeld S, Zhang J, Lee WP, Snipas SJ, Salvesen GS, Morris LX, Fitzgerald L, Zhang Y, Bertram EM, Goodnow CC, Dixit VM. Caspase-11 cleaves gasdermin D for non-canonical inflammasome signalling. Nature. 2015;526(7575):666–71.
pubmed: 26375259 doi: 10.1038/nature15541
Suzuki S, Franchi L, He Y, Muñoz-Planillo R, Mimuro H, Suzuki T, Sasakawa C, Núñez G. Shigella type III secretion protein MxiI is recognized by Naip2 to induce Nlrc4 inflammasome activation independently of pkcdelta. PLoS Pathog. 2014;10(2):e1003926.
pubmed: 24516390 pmcid: 3916413 doi: 10.1371/journal.ppat.1003926
Luchetti G, Roncaioli JL, Chavez RA, Schubert AF, Kofoed EM, Reja R, Cheung TK, Liang Y, Webster JD, Lehoux I, Skippington E, Reeder J, Haley B, Tan MW, Rose CM, Newton K, Kayagaki N, Vance RE, Dixit VM. Shigella ubiquitin ligase IpaH7.8 targets gasdermin D for degradation to prevent pyroptosis and enable infection. Cell Host Microbe. 2021;29(10):1521-30.e10.
Egan MS, Zhang J, Shin S. Human and mouse NAIP/NLRC4 inflammasome responses to bacterial infection. Curr Opin Microbiol. 2023;73:102298.
pubmed: 37058933 pmcid: 10225321 doi: 10.1016/j.mib.2023.102298
Bruchard M, Mignot G, Derangère V, Chalmin F, Chevriaux A, Végran F, Boireau W, Simon B, Ryffel B, Connat JL, Kanellopoulos J, Martin F, Rébé C, Apetoh L, Ghiringhelli F. Chemotherapy-triggered cathepsin B release in myeloid-derived suppressor cells activates the Nlrp3 inflammasome and promotes tumor growth. Nat Med. 2013;19(1):57–64.
pubmed: 23202296 doi: 10.1038/nm.2999
Biasizzo M, Kopitar-Jerala N. Interplay between NLRP3 inflammasome and autophagy. Front Immunol. 2020;11:591803.
pubmed: 33163006 pmcid: 7583715 doi: 10.3389/fimmu.2020.591803
Hsieh CY, Li LH, Rao YK, Ju TC, Nai YS, Chen YW, Hua KF. Mechanistic insight into the attenuation of gouty inflammation by Taiwanese green propolis via inhibition of the NLRP3 inflammasome. J Cell Physiol. 2019;234(4):4081–94.
pubmed: 30370562 doi: 10.1002/jcp.27204
Wu CH, Gan CH, Li LH, Chang JC, Chen ST, Menon MP, Cheng SM, Yang SP, Ho CL, Chernikov OV, Lin CH, Lam Y, Hua KF. A synthetic small molecule F240B decreases NLRP3 inflammasome activation by Autophagy Induction. Front Immunol. 2020;11:607564.
pubmed: 33424855 pmcid: 7793731 doi: 10.3389/fimmu.2020.607564
Wong WT, Li LH, Rao YK, Yang SP, Cheng SM, Lin WY, Cheng CC, Chen A, Hua KF. Repositioning of the β-Blocker carvedilol as a Novel Autophagy Inducer that inhibits the NLRP3 inflammasome. Front Immunol. 2018;9:1920.
pubmed: 30186288 pmcid: 6113403 doi: 10.3389/fimmu.2018.01920
Mantovani A, Sica A, Sozzani S, Allavena P, Vecchi A, Locati M. The chemokine system in diverse forms of macrophage activation and polarization. Trends Immunol. 2004;25(12):677–86.
pubmed: 15530839 doi: 10.1016/j.it.2004.09.015
Jaggi U, Yang M, Matundan HH, Hirose S, Shah PK, Sharifi BG, Ghiasi H. Increased phagocytosis in the presence of enhanced M2-like macrophage responses correlates with increased primary and latent HSV-1 infection. PLoS Pathog. 2020;16(10):e1008971.
pubmed: 33031415 pmcid: 7575112 doi: 10.1371/journal.ppat.1008971
Pang D, Huang Z, Li Q, Wang E, Liao S, Li E, Zou Y, Wang W. Antibacterial mechanism of Cinnamaldehyde: Modulation of Biosynthesis of Phosphatidylethanolamine and Phosphatidylglycerol in Staphylococcus aureus and Escherichia coli. J Agric Food Chem. 2021;69(45):13628–36.
pubmed: 34739242 doi: 10.1021/acs.jafc.1c04977
Akrami S, Amin M, Saki M. In vitro evaluation of the antibacterial effects of Cinnamomum zeylanicum essential oil against clinical multidrug-resistant Shigella isolates. Mol Biol Rep. 2021;48(3):2583–9.
pubmed: 33796990 doi: 10.1007/s11033-021-06309-w
Muhammad JS, Zaidi SF, Shaharyar S, Refaat A, Usmanghani K, Saiki I, Sugiyama T. Anti-inflammatory effect of cinnamaldehyde in Helicobacter pylori induced gastric inflammation. Biol Pharm Bull. 2015;38(1):109–15.
pubmed: 25744466 doi: 10.1248/bpb.b14-00609
Chung J, Kim S, Lee HA, Park MH, Kim S, Song YR, Na HS. Trans-cinnamic aldehyde inhibits Aggregatibacter actinomycetemcomitans-induced inflammation in THP-1-derived macrophages via autophagy activation. J Periodontol. 2018;89(10):1262–71.
pubmed: 29761921 doi: 10.1002/JPER.17-0727
Wang X, Jia Y, Wen L, Mu W, Wu X, Liu T, Liu X, Fang J, Luan Y, Chen P, Gao J, Nguyen KA, Cui J, Zeng G, Lan P, Chen Q, Cheng B, Wang Z. Porphyromonas gingivalis promotes colorectal carcinoma by activating the hematopoietic NLRP3 Inflammasome. Cancer Res. 2021;81(10):2745–59.
pubmed: 34003774 doi: 10.1158/0008-5472.CAN-20-3827
Beckwith KS, Beckwith MS, Ullmann S, Sætra RS, Kim H, Marstad A, Åsberg SE, Strand TA, Haug M, Niederweis M, Stenmark HA, Flo TH. Plasma membrane damage causes NLRP3 activation and pyroptosis during Mycobacterium tuberculosis infection. Nat Commun. 2020;11(1):2270.
pubmed: 32385301 pmcid: 7210277 doi: 10.1038/s41467-020-16143-6
Wang W, Hu D, Wu C, Feng Y, Li A, Liu W, Wang Y, Chen K, Tian M, Xiao F, Zhang Q, Shereen MA, Chen W, Pan P, Wan P, Wu K, Wu J. STING promotes NLRP3 localization in ER and facilitates NLRP3 deubiquitination to activate the inflammasome upon HSV-1 infection. PLoS Pathog. 2020;16(3):e1008335.
pubmed: 32187211 pmcid: 7080238 doi: 10.1371/journal.ppat.1008335
Hua KF, Yang FL, Chiu HW, Chou JC, Dong WC, Lin CN, Lin CY, Wang JT, Li LH, Chiu HW, Chiu YC, Wu SH. Capsular polysaccharide is involved in NLRP3 inflammasome activation by Klebsiella pneumoniae serotype K1. Infect Immun. 2015;83(9):3396–409.
pubmed: 26077758 pmcid: 4534678 doi: 10.1128/IAI.00125-15
Kim TW. Cinnamaldehyde induces autophagy-mediated cell death through ER stress and epigenetic modification in gastric cancer cells. Acta Pharmacol Sin. 2022;43(3):712–23.
pubmed: 33980998 doi: 10.1038/s41401-021-00672-x
Neto JGO, Boechat SK, Romão JS, Pazos-Moura CC, Oliveira KJ. Treatment with cinnamaldehyde reduces the visceral adiposity and regulates lipid metabolism, autophagy and endoplasmic reticulum stress in the liver of a rat model of early obesity. J Nutr Biochem. 2020;77:108321.
pubmed: 31869758 doi: 10.1016/j.jnutbio.2019.108321
Lee SC, Wang SY, Li CC, Liu CT. Anti-inflammatory effect of cinnamaldehyde and linalool from the leaf essential oil of Cinnamomum Osmophloeum Kanehira in endotoxin-induced mice. J Food Drug Anal. 2018;26(1):211–20.
pubmed: 29389558 doi: 10.1016/j.jfda.2017.03.006
Qu S, Shen Y, Wang M, Wang X, Yang Y. Suppression of miR-21 and miR-155 of macrophage by cinnamaldehyde ameliorates ulcerative colitis. Int Immunopharmacol. 2019;67:22–34.
pubmed: 30530166 doi: 10.1016/j.intimp.2018.11.045
Liu P, Wang J, Wen W, Pan T, Chen H, Fu Y, Wang F, Huang JH, Xu S. Cinnamaldehyde suppresses NLRP3 derived IL-1beta via activating succinate/HIF-1 in rheumatoid arthritis rats. Int Immunopharmacol. 2020;84:106570.
pubmed: 32413739 doi: 10.1016/j.intimp.2020.106570
Wang M, Yan S, Zhou Y, Xie P. Trans-cinnamaldehyde reverses depressive-like behaviors in chronic unpredictable mild stress rats by inhibiting NF-kappaB/NLRP3 inflammasome pathway. Evid Based Complement Alternat Med. 2020;2020:4572185.
pubmed: 32328132 pmcid: 7155764
Su SC, Hua KF, Lee H, Chao LK, Tan SK, Lee H, Yang SF, Hsu HY. LTA and LPS mediated activation of protein kinases in the regulation of inflammatory cytokines expression in macrophages. Clin Chim Acta. 2006;374(1–2):106–15.
pubmed: 16899235 doi: 10.1016/j.cca.2006.05.045
Kumari P, Kumar H. Dimensions of inflammation in host defense and diseases. Int Rev Immunol. 2022;41(1):1–3.
pubmed: 35044868 doi: 10.1080/08830185.2022.2014174

Auteurs

Kuo-Feng Hua (KF)

Department of Biotechnology and Animal Science, National Ilan University, Ilan, Taiwan.
Department of Medical Research, China Medical University Hospital, China Medical University, Taichung, Taiwan.

Yu-Bei Lin (YB)

Department of Biotechnology and Animal Science, National Ilan University, Ilan, Taiwan.
Department of Pathology, Tri-Service General Hospital, National Defense Medical Center, Taipei, Taiwan.

Hsiao-Wen Chiu (HW)

Department of Biotechnology and Animal Science, National Ilan University, Ilan, Taiwan.

Wei-Ting Wong (WT)

Department of Biotechnology and Animal Science, National Ilan University, Ilan, Taiwan.
Taiwan Autoantibody Biobank Initiative, Hualien Tzu Chi Hospital, Hualien, Taiwan.

Shuk-Man Ka (SM)

Graduate Institute of Aerospace and Undersea Medicine, Department of Medicine, National Defense Medical Center, Taipei, Taiwan.

Chun-Hsien Wu (CH)

Division of Cardiology, Department of Internal Medicine, Tri-Service General Hospital, National Defense Medical Center, Taipei, Taiwan.

Wen-Yu Lin (WY)

Department of Biotechnology and Animal Science, National Ilan University, Ilan, Taiwan.
Division of Cardiology, Department of Internal Medicine, Tri-Service General Hospital, National Defense Medical Center, Taipei, Taiwan.

Chien-Chun Wang (CC)

Infectious Disease Division, Linsen, Chinese Medicine and Kunming Branch, Taipei City Hospital, Taipei, Taiwan.
Kunming Prevention and Control Center, Taipei City Hospital, Taipei, Taiwan.

Chung-Hua Hsu (CH)

Linsen, Chinese Medicine and Kunming Branch, Taipei City Hospital, Taipei, Taiwan.
Institute of Traditional Medicine, School of Medicine, National Yang Ming Chiao Tung University, Taipei, Taiwan.

Hsien-Ta Hsu (HT)

Division of Neurosurgery, Taipei Tzu Chi Hospital, Buddhist Tzu Chi Medical Foundation, New Taipei City, Taiwan.
School of Medicine, Buddhist Tzu Chi University, Hualien, Taiwan.

Chen-Lung Ho (CL)

Division of Wood Cellulose, Taiwan Forestry Research Institute, Taipei, Taiwan.

Lan-Hui Li (LH)

Department of Laboratory Medicine, Linsen, Chinese Medicine and Kunming Branch, Taipei City Hospital, Taipei, Taiwan. A1525@tpech.gov.tw.

Classifications MeSH