, Volume 23, Issue 5–6, pp 314–328 | Cite as

Prodigiosin stimulates endoplasmic reticulum stress and induces autophagic cell death in glioblastoma cells

  • Shu-Yu Cheng
  • Nan-Fu Chen
  • Hsiao-Mei Kuo
  • San-Nan Yang
  • Chun-Sung Sung
  • Ping-Jyun Sung
  • Zhi-Hong Wen
  • Wu-Fu Chen


Prodigiosin, a secondary metabolite isolated from marine Vibrio sp., has antimicrobial and anticancer properties. This study investigated the cell death mechanism of prodigiosin in glioblastoma. Glioblastoma multiforme (GBM) is an aggressive primary cancer of the central nervous system. Despite treatment, or standard therapy, the median survival of glioblastoma patients is about 14.6 month. The results of the present study clearly showed that prodigiosin significantly reduced the cell viability and neurosphere formation ability of U87MG and GBM8401 human glioblastoma cell lines. Moreover, prodigiosin with fluorescence signals was detected in the endoplasmic reticulum and found to induce excessive levels of autophagy. These findings were confirmed by observation of LC3 puncta formation and acridine orange staining. Furthermore, prodigiosin caused cell death by activating the JNK pathway and decreasing the AKT/mTOR pathway in glioblastoma cells. Moreover, we found that the autophagy inhibitor 3-methyladenine reversed prodigiosin induced autophagic cell death. These findings of this study suggest that prodigiosin induces autophagic cell death and apoptosis in glioblastoma cells.


Prodigiosin Autophagic cell death Glioblastoma 



This research was supported by Ministry of Science and Technology, Taiwan (105-2320-B-182A-015), National Research Program for Biopharmaceuticals, Taiwan (105-2325-B-110-001), and Chang Gung Memorial Hospital, Taiwan (CMRPG8D0821).

Supplementary material

10495_2018_1456_MOESM1_ESM.docx (959 kb)
Supplementary material 1 (DOCX 958 KB)
10495_2018_1456_MOESM2_ESM.xlsx (35 kb)
Supplementary material 2 (XLSX 35 KB)


  1. 1.
    Darshan N, Manonmani H (2015) Prodigiosin and its potential applications. J Food Sci Technol 52(9):5393–5407CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Lipinski CA, Lombardo F, Dominy BW, Feeney PJ (1997) Experimental and computational approaches to estimate solubility and permeability in drug discovery and development settings. Adv Drug Deliv Rev 23(1–3):3–25. CrossRefGoogle Scholar
  3. 3.
    Campas C, Dalmau M, Montaner B, Barragan M, Bellosillo B, Colomer D, Pons G, Pérez-Tomás R, Gil J (2003) Prodigiosin induces apoptosis of B and T cells from B-cell chronic lymphocytic leukemia. Leukemia 17(4):746–750. CrossRefPubMedGoogle Scholar
  4. 4.
    Espona-Fiedler M, Soto-Cerrato V, Hosseini A, Lizcano J, Guallar V, Quesada R, Gao T, Pérez-Tomás R (2012) Identification of dual mTORC1 and mTORC2 inhibitors in melanoma cells: prodigiosin vs. obatoclax. Biochem Pharmacol 83(4):489–496. CrossRefPubMedGoogle Scholar
  5. 5.
    Montaner B, Navarro S, Piqué M, Vilaseca M, Martinell M, Giralt E, Gil J, Pérez-Tomás R (2000) Prodigiosin from the supernatant of Serratia marcescens induces apoptosis in haematopoietic cancer cell lines. Br J Pharmacol 131(3):585–593. CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Wang Z, Li B, Zhou L, Yu S, Su Z, Song J, Sun Q, Sha O, Wang X, Jiang W (2016) Prodigiosin inhibits Wnt/β-catenin signaling and exerts anticancer activity in breast cancer cells. Proc Natl Acad Sci 113(46):13150–13155. CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Prabhu VV, Hong B, Allen JE, Zhang S, Lulla AR, Dicker DT, El-Deiry WS (2016) Small-molecule prodigiosin restores p53 tumor suppressor activity in chemoresistant colorectal cancer stem cells via c-Jun-mediated ∆Np73 inhibition and p73 activation. Cancer Res 76(7):1989–1999. CrossRefPubMedGoogle Scholar
  8. 8.
    Tomás RP, Ruir CD, Montaner B (2001) Prodigiosin induces cell death and morphological changes indicative of apoptosis in gastric cancer cell line HGT-1. Histol Histopathol 16(2):415–421. CrossRefGoogle Scholar
  9. 9.
    Ramirez YP, Weatherbee JL, Wheelhouse RT, Ross AH (2013) Glioblastoma multiforme therapy and mechanisms of resistance. Pharmaceuticals 6(12):1475–1506. CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Stupp R, Hegi ME, Mason WP, van den Bent MJ, Taphoorn MJ, Janzer RC, Ludwin SK, Allgeier A, Fisher B, Belanger K (2009) Effects of radiotherapy with concomitant and adjuvant temozolomide versus radiotherapy alone on survival in glioblastoma in a randomised phase III study: 5-year analysis of the EORTC-NCIC trial. Lancet Oncol 10(5):459–466. CrossRefPubMedGoogle Scholar
  11. 11.
    Oliva CR, Nozell SE, Diers A, McClugage SG, Sarkaria JN, Markert JM, Darley-Usmar VM, Bailey SM, Gillespie GY, Landar A (2010) Acquisition of temozolomide chemoresistance in gliomas leads to remodeling of mitochondrial electron transport chain. J Biol Chem 285(51):39759–39767. CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Hatanpaa KJ, Burma S, Zhao D, Habib AA (2010) Epidermal growth factor receptor in glioma: signal transduction, neuropathology, imaging, and radioresistance. Neoplasia 12(9):675–684. CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Yang Y, Shao N, Luo G, Li L, Zheng L, Nilsson-Ehle P, Xu N (2010) Mutations of PTEN gene in gliomas correlate to tumor differentiation and short-term survival rate. Anticancer Res 30(3):981–985. PubMedCrossRefGoogle Scholar
  14. 14.
    Taylor E, Furnari TB, Cavenee FK W (2012) Targeting EGFR for treatment of glioblastoma: molecular basis to overcome resistance. Curr Cancer Drug Targets 12(3):197–209. CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Annovazzi L, Mellai M, Caldera V, Valente G, Tessitore L, Schiffer D (2009) mTOR, S6 and AKT expression in relation to proliferation and apoptosis/autophagy in glioma. Anticancer Res 29(8):3087–3094. PubMedCrossRefGoogle Scholar
  16. 16.
    Rahaman SO, Harbor PC, Chernova O, Barnett GH, Vogelbaum MA, Haque SJ (2002) Inhibition of constitutively active Stat3 suppresses proliferation and induces apoptosis in glioblastoma multiforme cells. Oncogene 21(55):8404–8413. CrossRefPubMedGoogle Scholar
  17. 17.
    Senft D, Ze’ev AR (2015) UPR, autophagy, and mitochondria crosstalk underlies the ER stress response. Trends Biochem Sci 40 (3):141–148. CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Martinez-Outschoorn UE, Whitaker-Menezes D, Pavlides S, Chiavarina B, Bonuccelli G, Trimmer C, Tsirigos A, Migneco G, Witkiewicz AK, Balliet RM (2010) The autophagic tumor stroma model of cancer or “battery-operated tumor growth” a simple solution to the autophagy paradox. Cell cycle 9(21):4297–4306. CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Eisenberg-Lerner A, Bialik S, Simon H-U, Kimchi A (2009) Life and death partners: apoptosis, autophagy and the cross-talk between them. Cell Death Differ 16(7):966–975. CrossRefPubMedGoogle Scholar
  20. 20.
    Tsujimoto Y, Shimizu S (2005) Another way to die: autophagic programmed cell death. Cell Death Differ 12:1528–1534. CrossRefPubMedGoogle Scholar
  21. 21.
    Sano R, Reed JC (2013) ER stress-induced cell death mechanisms. Biochim Biophys Acta 1833(12):3460–3470. CrossRefPubMedGoogle Scholar
  22. 22.
    Oyadomari S, Mori M (2004) Roles of CHOP/GADD153 in endoplasmic reticulum stress. Cell Death Differ 11(4):381–389. CrossRefPubMedGoogle Scholar
  23. 23.
    Jang G-H, Lee M (2014) BH3-mimetic gossypol-induced autophagic cell death in mutant BRAF melanoma cells with high expression of p21 Cip1. Life Sci 102(1):41–48. CrossRefPubMedGoogle Scholar
  24. 24.
    Li J-R, Cheng C-L, Yang W-J, Yang C-R, Ou Y-C, Wu M-J, Ko J-L (2014) FIP-gts potentiate autophagic cell death against cisplatin-resistant urothelial cancer cells. Anticancer Res 34(6):2973–2983. PubMedCrossRefGoogle Scholar
  25. 25.
    Sui X, Chen R, Wang Z, Huang Z, Kong N, Zhang M, Han W, Lou F, Yang J, Zhang Q (2013) Autophagy and chemotherapy resistance: a promising therapeutic target for cancer treatment. Cell Death Dis 4(10):e838. CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Lee WH, Yeh MY, Tu YC, Han SH, Wang YC (1988) Establishment and characterization of a malignant glioma cell line, GBM8401/TSGH, NDMC. J Surg Oncol 38(3):173–181. CrossRefPubMedGoogle Scholar
  27. 27.
    Hong X, Chedid K, Kalkanis SN (2012) Glioblastoma cell line-derived spheres in serum‑containing medium versus serum-free medium: a comparison of cancer stem cell properties. Int J Oncol 41(5):1693–1700. CrossRefPubMedGoogle Scholar
  28. 28.
    Kimura S, Noda T, Yoshimori T (2007) Dissection of the autophagosome maturation process by a novel reporter protein, tandem fluorescent-tagged LC3. Autophagy 3(5):452–460. CrossRefPubMedGoogle Scholar
  29. 29.
    Panosyan EH, Laks DR, Masterman-Smith M, Mottahedeh J, Yong WH, Cloughesy TF, Lazareff JA, Mischel PS, Moore TB, Kornblum HI (2010) Clinical outcome in pediatric glial and embryonal brain tumors correlates with in vitro multi-passageable neurosphere formation. Pediatr Blood Cancer 55(4):644–651. CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Müller-Taubenberger A, Lupas AN, Li H, Ecke M, Simmeth E, Gerisch G (2001) Calreticulin and calnexin in the endoplasmic reticulum are important for phagocytosis. EMBO J 20(23):6772–6782CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    Dunn KW, Kamocka MM, McDonald JH (2011) A practical guide to evaluating colocalization in biological microscopy. Am J Physiol-Cell Physiol 300(4):C723–C742CrossRefGoogle Scholar
  32. 32.
    Evans JD (1996) Straightforward statistics for the behavioral sciences. Brooks/Cole, Pacific GroveGoogle Scholar
  33. 33.
    van Schadewijk A, van’t Wout EF, Stolk J, Hiemstra PS (2012) A quantitative method for detection of spliced X-box binding protein-1 (XBP1) mRNA as a measure of endoplasmic reticulum (ER) stress. Cell Stress Chaperones 17(2):275–279. CrossRefPubMedGoogle Scholar
  34. 34.
    Li J, Ni M, Lee B, Barron E, Hinton D, Lee A (2008) The unfolded protein response regulator GRP78/BiP is required for endoplasmic reticulum integrity and stress-induced autophagy in mammalian cells. Cell Death Differ 15(9):1460–1471. CrossRefPubMedPubMedCentralGoogle Scholar
  35. 35.
    Liu K, Shi Y, Guo X, Wang S, Ouyang Y, Hao M, Liu D, Qiao L, Li N, Zheng J (2014) CHOP mediates ASPP2-induced autophagic apoptosis in hepatoma cells by releasing Beclin-1 from Bcl-2 and inducing nuclear translocation of Bcl-2. Cell Death Dis 5(7):e1323. CrossRefPubMedPubMedCentralGoogle Scholar
  36. 36.
    Kabeya Y, Mizushima N, Ueno T, Yamamoto A, Kirisako T, Noda T, Kominami E, Ohsumi Y, Yoshimori T (2000) LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing. EMBO J 19(21):5720–5728. CrossRefPubMedPubMedCentralGoogle Scholar
  37. 37.
    Shimizu S, Konishi A, Nishida Y, Mizuta T, Nishina H, Yamamoto A, Tsujimoto Y (2010) Involvement of JNK in the regulation of autophagic cell death. Oncogene 29(14):2070–2082CrossRefPubMedGoogle Scholar
  38. 38.
    Eberhart K, Oral O, Gozuacik D (2013) Induction of autophagic cell death by anticancer agents. Autophagy. CrossRefGoogle Scholar
  39. 39.
    Nihira K, Miki Y, Ono K, Suzuki T, Sasano H (2014) An inhibition of p62/SQSTM1 caused autophagic cell death of several human carcinoma cells. Cancer Sci 105(5):568–575. CrossRefPubMedPubMedCentralGoogle Scholar
  40. 40.
    Karch J, Schips TG, Maliken BD, Brody MJ, Sargent MA, Kanisciak O, Molkentin JD (2017) Autophagic cell death is dependent on lysosomal membrane permeability through Bax and Bak. eLife. PubMedPubMedCentralCrossRefGoogle Scholar
  41. 41.
    Liu Y, Levine B (2015) Autosis and autophagic cell death: the dark side of autophagy. Cell Death Differ 22(3):367–376. CrossRefPubMedGoogle Scholar
  42. 42.
    Lichstein HC, Van De Sand VF (1946) The antibiotic activity of violacein, prodigiosin, and phthiocol. J Bacteriol 52(1):145. PubMedCentralCrossRefPubMedGoogle Scholar
  43. 43.
    Hosseini A, Espona-Fiedler M, Soto-Cerrato V, Quesada R, Pérez-Tomás R, Guallar V (2013) Molecular interactions of prodiginines with the BH3 domain of anti-apoptotic Bcl-2 family members. PLoS ONE 8(2):e57562. CrossRefPubMedPubMedCentralGoogle Scholar
  44. 44.
    Zhang J, Shen Y, Liu J, Wei D (2005) Antimetastatic effect of prodigiosin through inhibition of tumor invasion. Biochem Pharmacol 69(3):407–414. CrossRefPubMedGoogle Scholar
  45. 45.
    Soto-Cerrato V, Viñals F, Lambert JR, Pérez-Tomás R (2007) The anticancer agent prodigiosin induces p21 WAF1/CIP1 expression via transforming growth factor-beta receptor pathway. Biochem Pharmacol 74(9):1340–1349. CrossRefPubMedGoogle Scholar
  46. 46.
    Alison MR, Lim SM, Nicholson LJ (2011) Cancer stem cells: problems for therapy? J Pathol 223(2):148–162. CrossRefGoogle Scholar
  47. 47.
    Chen Y-C, Ingram PN, Fouladdel S, McDermott SP, Azizi E, Wicha MS, Yoon E (2016) High-throughput single-cell derived sphere formation for cancer stem-like cell identification and analysis. Sci Rep. CrossRefPubMedPubMedCentralGoogle Scholar
  48. 48.
    Ojha R, Bhattacharyya S, Singh SK (2015) Autophagy in cancer stem cells: a potential link between chemoresistance, recurrence, and metastasis. BioRes Open Access 4(1):97–108. CrossRefPubMedPubMedCentralGoogle Scholar
  49. 49.
    Jiang H, Gomez-Manzano C, Aoki H, Alonso MM, Kondo S, McCormick F, Xu J, Kondo Y, Bekele BN, Colman H (2007) Examination of the therapeutic potential of Delta-24-RGD in brain tumor stem cells: role of autophagic cell death. J Natl Cancer Inst 99(18):1410–1414. CrossRefPubMedGoogle Scholar
  50. 50.
    Kanzawa T, Germano I, Komata T, Ito H, Kondo Y, Kondo S (2004) Role of autophagy in temozolomide-induced cytotoxicity for malignant glioma cells. Cell Death Differ 11(4):448–457. CrossRefPubMedGoogle Scholar
  51. 51.
    Tsukada M, Ohsumi Y (1993) Isolation and characterization of autophagy-defective mutants of Saccharomyces cerevisiae. FEBS Lett 333(1–2):169–174. CrossRefPubMedGoogle Scholar
  52. 52.
    Bernales S, Schuck S, Walter P (2007) ER-phagy: selective autophagy of the endoplasmic reticulum. Autophagy 3(3):285–287. CrossRefPubMedGoogle Scholar
  53. 53.
    Degenhardt K, Mathew R, Beaudoin B, Bray K, Anderson D, Chen G, Mukherjee C, Shi Y, Gélinas C, Fan Y (2006) Autophagy promotes tumor cell survival and restricts necrosis, inflammation, and tumorigenesis. Cancer Cell 10(1):51–64. CrossRefPubMedPubMedCentralGoogle Scholar
  54. 54.
    McLendon R, Friedman A, Bigner D, Van Meir EG, Brat DJ, Mastrogianakis GM, Olson JJ, Mikkelsen T, Lehman N, Aldape K (2008) Comprehensive genomic characterization defines human glioblastoma genes and core pathways. Nature 455(7216):1061–1068. CrossRefGoogle Scholar
  55. 55.
    Carico C, Nuño M, Mukherjee D, Elramsisy A, Dantis J, Hu J, Rudnick J, John SY, Black KL, Bannykh SI (2012) Loss of PTEN is not associated with poor survival in newly diagnosed glioblastoma patients of the temozolomide era. PLoS ONE 7(3):e33684. CrossRefPubMedPubMedCentralGoogle Scholar
  56. 56.
    Gozuacik D, Kimchi A (2004) Autophagy as a cell death and tumor suppressor mechanism. Oncogene 23(16):2891–2906. CrossRefPubMedGoogle Scholar
  57. 57.
    Fantus D, Rogers NM, Grahammer F, Huber TB, Thomson AW (2016) Roles of mTOR complexes in the kidney: implications for renal disease and transplantation. Nat Rev Nephrol. PubMedCrossRefGoogle Scholar
  58. 58.
    Gozuacik D, Kimchi A (2007) Autophagy and cell death. Curr Top Dev Biol 78:217–245. CrossRefPubMedGoogle Scholar
  59. 59.
    Schweichel JU, Merker HJ (1973) The morphology of various types of cell death in prenatal tissues. Teratology 7(3):253–266. CrossRefPubMedGoogle Scholar
  60. 60.
    Eberhart K, Oral O, Gozuacik D (2013) Autophagy: Chap. 13. Induction of autophagic cell death by anticancer agents. Elsevier Inc. Chapters, New YorkGoogle Scholar
  61. 61.
    Ogata M, Hino S-i, Saito A, Morikawa K, Kondo S, Kanemoto S, Murakami T, Taniguchi M, Tanii I, Yoshinaga K (2006) Autophagy is activated for cell survival after endoplasmic reticulum stress. Mol Cell Biol 26(24):9220–9231. CrossRefPubMedPubMedCentralGoogle Scholar
  62. 62.
    Leppä S, Bohmann D (1999) Diverse functions of JNK signaling and c-Jun in stress response and apoptosis. Oncogene.
  63. 63.
    Wei Y, Pattingre S, Sinha S, Bassik M, Levine B (2008) JNK1-mediated phosphorylation of Bcl-2 regulates starvation-induced autophagy. Molecular Cell 30(6):678–688. CrossRefPubMedPubMedCentralGoogle Scholar
  64. 64.
    Vegliante R, Desideri E, Di Leo L, Ciriolo MR (2016) Dehydroepiandrosterone triggers autophagic cell death in human hepatoma cell line HepG2 via JNK-mediated p62/SQSTM1 expression. Carcinogenesis. PubMedCrossRefGoogle Scholar
  65. 65.
    Zhang C, Jia X, Wang K, Bao J, Li P, Chen M, Wan J-B, Su H, Mei Z, He C (2016) Polyphyllin VII induces an autophagic cell death by activation of the JNK pathway and inhibition of PI3K/AKT/mTOR pathway in HepG2 cells. PLoS ONE 11(1):e0147405. CrossRefPubMedPubMedCentralGoogle Scholar
  66. 66.
    Louis DN, Von Deimling A, Chung RY, Rubio M-P, Whaley JM, Eibl RH, Ohgaki H, Wiestler OD, Thor AD, Seizinger BR (1993) Comparative study of p53 gene and protein alterations in human astrocytic tumors. J Neuropathol Exp Neurol 52(1):31–38. CrossRefPubMedGoogle Scholar
  67. 67.
    Strano S, Dell’Orso S, Di Agostino S, Fontemaggi G, Sacchi A, Blandino G (2007) Mutant p53: an oncogenic transcription factor. Oncogene 26(15):2212–2219. CrossRefPubMedGoogle Scholar
  68. 68.
    Muller PA, Vousden KH (2014) Mutant p53 in cancer: new functions and therapeutic opportunities. Cancer Cell 25(3):304–317. CrossRefPubMedPubMedCentralGoogle Scholar
  69. 69.
    Wang X, Chen J-x, Liu J-p, You C, Liu Y-h, Mao Q (2014) Gain of function of mutant TP53 in glioblastoma: prognosis and response to temozolomide. Ann Surg Oncol 21(4):1337–1344. CrossRefPubMedGoogle Scholar
  70. 70.
    Cordani M, Butera G, Pacchiana R, Donadelli M (2016) Molecular interplay between mutant p53 proteins and autophagy in cancer cells. Biochim Biophys Acta. PubMedCrossRefGoogle Scholar
  71. 71.
    Hong B, Prabhu VV, Zhang S, van den Heuvel APJ, Dicker DT, Kopelovich L, El-Deiry WS (2014) Prodigiosin rescues deficient p53 signaling and antitumor effects via upregulating p73 and disrupting its interaction with mutant p53. Cancer Res 74(4):1153–1165. CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Shu-Yu Cheng
    • 1
    • 2
    • 3
  • Nan-Fu Chen
    • 4
    • 5
  • Hsiao-Mei Kuo
    • 3
    • 6
    • 10
  • San-Nan Yang
    • 7
  • Chun-Sung Sung
    • 8
    • 9
  • Ping-Jyun Sung
    • 10
    • 11
    • 12
  • Zhi-Hong Wen
    • 1
    • 3
    • 10
  • Wu-Fu Chen
    • 10
    • 13
    • 14
  1. 1.Doctoral Degree Program in Marine BiotechnologyNational Sun Yat-Sen UniversityKaohsiungTaiwan, Republic of China
  2. 2.Doctoral Degree Program in Marine BiotechnologyAcademia SinicaTaipei CityTaiwan, Republic of China
  3. 3.Marine Biomedical Laboratory & Center for Translational Biopharmaceuticals, Department of Marine Biotechnology and ResourcesNational Sun Yat-sen UniversityKaohsiungTaiwan, Republic of China
  4. 4.Division of Neurosurgery, Department of SurgeryKaohsiung Armed Forces General HospitalKaohsiungTaiwan, Republic of China
  5. 5.Department of Neurological Surgery, Tri-Service General HospitalNational Defense Medical CenterTaipei CityTaiwan, Republic of China
  6. 6.Center for NeuroscienceNational Sun Yat-Sen UniversityKaohsiungTaiwan, Republic of China
  7. 7.Department of Pediatrics, E-DA Hospital, School of Medicine, College of MedicineI-SHOU UniversityKaohsiungTaiwan, Republic of China
  8. 8.Department of AnesthesiologyTaipei Veterans General HospitalTaipei CityTaiwan, Republic of China
  9. 9.School of MedicineNational Yang-Ming UniversityTaipei CityTaiwan, Republic of China
  10. 10.Department of Marine Biotechnology and ResourcesNational Sun Yat-sen UniversityKaohsiungTaiwan, Republic of China
  11. 11.Graduate Institute of Marine BiologyNational Dong Hwa UniversityChecheng TownshipTaiwan, Republic of China
  12. 12.National Museum of Marine Biology and AquariumChecheng TownshipTaiwan, Republic of China
  13. 13.Department of NeurosurgeryKaohsiung Chang Gung Memorial Hospital and Chang Gung University College of MedicineKaohsiungTaiwan, Republic of China
  14. 14.Department of NeurosurgeryXiamen Chang Gung HospitalXiamen ShiChina

Personalised recommendations