TH+ cells 的數量都有增加，因此我們想知道在經過治療後，血清 中多巴胺的表現量是否有變化。從行為模式分析的數據我們可以 發現，在第 9 天，行為就有回復的現象。所以我們取了第 9 天以 及第 23 天的血清來做 DA 的 ELISA 分析。根據 ELISA 的結果 (Figure 14) ，在第 9 天的 MPTP group，其 DA 的表現量有顯著的 下降；而經過治療的組別(cells group, cells+BP group 以及 BP group)，DA 的量就已經回復到接近於正常表現的水平。第 23 天 的數據顯示，MPTP group 的 DA 表現量也逐漸回復到接近正常 表現的水平；而經過治療的組別((cells group, cells+BP group 以及 BP group)，DA 的量依舊維持表現。
9. 以 TUNEL assay 探討短時間(5 天)細胞移殖療法的機制
根據先前的研究，MPTP 會引發 SN 的細胞凋亡[11]。因此，
我們利用 TUNEL assay 來觀察經過治療後的變化。一般未經過處 理的組別，僅有些許的 TUNEL 表現(Figure 15A)，而經過 MPTP 處理過的組別，TUNEL 大量表現(Figure 15B)；在經過治療組別：
cells group (Figure 15C), cells+BP group (Figure 15D)以及 BP group (Figure 15E)都可以看見 TUNEL 的表現有顯著的下降，特別是 cells+BP group。
伍、討論
間葉幹細胞[44]、intracerebral peripheral blood stem cells (CD34+)[45]
以及嗅鞘細胞[46]。我們所使用的iPS-OSH derived neuronal stem cell 可以改善行為模式的數據(Figure 3、Figure 4) ，並且分化成為神經 (Figure 6)；取得的方式也比骨髓幹細胞以及intracerebral peripheral blood stem cells (CD34+)來得方便，在未來的臨床運用上有很大的效 力。 為 alpha-synuclein 蛋白錯誤摺疊而導致的 PD 有些許的不同[51]；本 實驗室從美國 Jackson Laboratory 引進一種帶有人類 synuclein, alpha
細胞來治療帕金森氏症：嗅鞘細胞[54, 55]，嗅鞘細胞在於神經損傷 的治療方面都有不錯的成效[56, 57]。我們所使用的 iPS-OSH derived neuronal stem cell 對於帕金森氏症的治療在細胞移植後 5 天就可以發 現行為模式的改善(Figure 5)，在 SN 區多巴胺神經的數量也有明顯增 加(Figure 8、Figure 10)，並且降低的細胞凋亡的發生(Figure 15)。
實驗的數據顯示，我們實驗室所製造的iPS-OSH對缺氧性中風以 及帕金森氏症都有治療的功效，有作為未來細胞移植臨床治療的來 源。
陸、結論
本實驗室製造出不帶致癌基因以及非病毒轉染的誘導性多功能 幹細胞具有分化成為神經幹細胞的能力，在移殖進缺氧性中風小鼠的 模式後，我們分化的神經幹細胞可以再分化成為中樞神經系統類的神 經細胞，並且回復缺氧性中風小鼠的行為能力；在移殖進帕金森氏症 小鼠的模式後，我們分化的神經幹細胞可以再分化成為多巴胺神經、
神經膠質細胞以及分化中的神經細胞，並且可以降低Iba1的表現、增 加神經新生KI67的表現以及TH的表現量，並降低SN區細胞凋亡的現 象，回復帕金森氏症小鼠的行為能力。
柒、參考文獻
1. Chiu L, Pai L, Shyu WC, Jayne Chen TR, Chang TP: Analysis of costs borne by families of patients hospitalized for stroke. Zhonghua Yi Xue Za Zhi (Taipei) 1998, 61(5):267-275.
2. Hinkle JL, Guanci MM: Acute ischemic stroke review. The Journal of neuroscience nursing : journal of the American Association of Neuroscience Nurses 2007, 39(5):285-293, 310.
3. Chopp M, Li Y: Treatment of stroke and intracerebral hemorrhage with cellular and pharmacological restorative therapies. Acta neurochirurgica Supplement 2008, 105:79-83.
4. Antithrombotic Trialists C: Collaborative meta-analysis of randomised trials of antiplatelet therapy for prevention of death, myocardial infarction, and stroke in high risk patients. BMJ 2002, 324(7329):71-86.
5. 張谷州: 台灣急性缺血性腦中風的靜脈注射血栓溶解劑治療回顧. 腦
中風會訊 2000, 7(2):5-5.
6. Parkinson J: An essay on the shaking palsy. 1817. J Neuropsychiatry Clin Neurosci 2002, 14(2):223-236; discussion 222.
7. Barbosa ER, Limongi JC, Cummings JL: Parkinson's disease. Psychiatr Clin North Am 1997, 20(4):769-790.
8. Mizuno Y, Hattori N, Matsumine H: Neurochemical and neurogenetic correlates of Parkinson's disease. Journal of neurochemistry 1998, 71(3):893-902.
9. Adams JD, Jr., Chang ML, Klaidman L: Parkinson's disease--redox mechanisms. Current medicinal chemistry 2001, 8(7):809-814.
10. Langston JW: MPTP and Parkinson's disease. TINS 1985.
11. Blum D, Torch S, Lambeng N, Nissou M, Benabid AL, Sadoul R, Verna JM:
Molecular pathways involved in the neurotoxicity of 6-OHDA, dopamine and MPTP: contribution to the apoptotic theory in Parkinson's disease.
Prog Neurobiol 2001, 65(2):135-172.
12. Munoz P, Huenchuguala S, Paris I, Segura-Aguilar J: Dopamine oxidation and autophagy. Parkinsons Dis 2012, 2012:920953.
13. Lefrère JJ, Berche P: La thérapeutique du docteur Brown-Séquard. Ann Endocrinol (Paris) 2010, 71(2):69-75.
14. Kemeny DM, MacAry PA: Immunology: making magic bullets. BMJ 2007,
334 Suppl 1:s13.
15. Becker AJ, Mc CE, Till JE: Cytological demonstration of the clonal nature of spleen colonies derived from transplanted mouse marrow cells. Nature 1963, 197:452-454.
16. Siminovitch L, McCulloch EA, Till JE: THE DISTRIBUTION OF COLONY-FORMING CELLS AMONG SPLEEN COLONIES. J Cell Physiol 1963, 62:327-336.
17. Takahashi K, Yamanaka S: Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 2006, 126(4):663-676.
18. Takahashi K, Tanabe K, Ohnuki M, Narita M, Ichisaka T, Tomoda K, Yamanaka S: Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell 2007, 131(5):861-872.
19. Rhee YH, Ko JY, Chang MY, Yi SH, Kim D, Kim CH, Shim JW, Jo AY, Kim BW, Lee H et al: Protein-based human iPS cells efficiently generate
functional dopamine neurons and can treat a rat model of Parkinson disease. The Journal of clinical investigation 2011, 121(6):2326-2335.
20. Weir GC, Cavelti-Weder C, Bonner-Weir S: Stem cell approaches for diabetes: towards beta cell replacement. Genome Med 2011, 3(9):61.
21. Liu SP, Fu RH, Huang YC, Chen SY, Chien YJ, Hsu CY, Tsai CH, Shyu WC, Lin SZ: Induced pluripotent stem (iPS) cell research overview. Cell
transplantation 2011, 20(1):15-19.
22. Reynolds BA, Weiss S: Generation of neurons and astrocytes from isolated cells of the adult mammalian central nervous system. Science 1992,
255(5052):1707-1710.
23. Esmaeilzade B, Nobakht M, Joghataei MT, Rahbar Roshandel N, Rasouli H, Samadi Kuchaksaraei A, Hosseini SM, Najafzade N, Asalgoo S, Hejazian LB et al: Delivery of epidermal neural crest stem cells (EPI-NCSC) to
hippocamp in Alzheimer's disease rat model. Iranian biomedical journal 2012, 16(1):1-9.
24. Arias-Carrion O, Yuan TF: Autologous neural stem cell transplantation: a new treatment option for Parkinson's disease? Med Hypotheses 2009,
enhance amelioration of ischemic stroke in mice. Stroke; a journal of cerebral circulation 2012, 43(9):2423-2429.
26. Nakayama T, Momoki-Soga T, Yamaguchi K, Inoue N: Efficient production of neural stem cells and neurons from embryonic stem cells. Neuroreport 2004, 15(3):487-491.
27. Lie KH, Chung HC, Sidhu KS: Derivation, propagation, and
characterization of neuroprogenitors from pluripotent stem cells (hESCs and hiPSCs). Methods Mol Biol 2012, 873:237-246.
28. Tsuji O, Miura K, Fujiyoshi K, Momoshima S, Nakamura M, Okano H: Cell therapy for spinal cord injury by neural stem/progenitor cells derived from iPS/ES cells. Neurotherapeutics : the journal of the American Society for Experimental NeuroTherapeutics 2011, 8(4):668-676.
29. Dahlstrand J, Lardelli M, Lendahl U: Nestin mRNA expression correlates with the central nervous system progenitor cell state in many, but not all, regions of developing central nervous system. Brain Res Dev Brain Res 1995, 84(1):109-129.
30. Sansom SN, Griffiths DS, Faedo A, Kleinjan DJ, Ruan Y, Smith J, van Heyningen V, Rubenstein JL, Livesey FJ: The level of the transcription factor Pax6 is essential for controlling the balance between neural stem cell self-renewal and neurogenesis. PLoS Genet 2009, 5(6):e1000511.
31. Wang H, Chen R, Xu H: [Chemical constituents of radix Angelicae Sinensis]. Zhongguo Zhong Yao Za Zhi 1998, 23(3):167-168, inside backcover.
32. Tsai NM, Lin SZ, Lee CC, Chen SP, Su HC, Chang WL, Harn HJ: The antitumor effects of Angelica sinensis on malignant brain tumors in vitro and in vivo. Clin Cancer Res 2005, 11(9):3475-3484.
33. Tsai NM, Chen YL, Lee CC, Lin PC, Cheng YL, Chang WL, Lin SZ, Harn HJ:
The natural compound n-butylidenephthalide derived from Angelica sinensis inhibits malignant brain tumor growth in vitro and in vivo.
Journal of neurochemistry 2006, 99(4):1251-1262.
34. Liu SP, Harn HJ, Chien YJ, Chang CH, Hsu CY, Fu RH, Huang YC, Chen SY, Shyu WC, Lin SZ: n-Butylidenephthalide (BP) maintains stem cell
pluripotency by activating Jak2/Stat3 pathway and increases the efficiency of iPS cells generation. PLoS One 2012, 7(9):e44024.
35. Bae N, Ahn T, Chung S, Oh MS, Ko H, Oh H, Park G, Yang HO: The
neuroprotective effect of modified Yeoldahanso-tang via autophagy enhancement in models of Parkinson's disease. J Ethnopharmacol 2011, 134(2):313-322.
36. Jerry JB: Methods of Behavior Analysis in Neuroscience second edition.
2009.
37. Saijo K, Glass CK: Microglial cell origin and phenotypes in health and disease. Nat Rev Immunol 2011, 11(11):775-787.
38. Block ML, Zecca L, Hong JS: Microglia-mediated neurotoxicity:
uncovering the molecular mechanisms. Nat Rev Neurosci 2007, 8(1):57-69.
39. 林明政, 李名世, 王俊民, 梁雅娟, 陳甫州: 雷射杜卜勒微流儀之原理
與臨床醫學之應用. 中華民國醫檢會報 2002.
40. Mah S, Deveber G, Wei XC, Liapounova N, Kirton A: Cerebellar Atrophy in Childhood Arterial Ischemic Stroke: Acute Diffusion MRI Biomarkers.
Stroke; a journal of cerebral circulation 2013.
41. Johnson MM: Stroke and CT Perfusion. Radiol Technol 2012, 83(5):467CT-486CT.
42. Kawai H, Yamashita T, Ohta Y, Deguchi K, Nagotani S, Zhang X, Ikeda Y, Matsuura T, Abe K: Tridermal tumorigenesis of induced pluripotent stem cells transplanted in ischemic brain. Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism 2010, 30(8):1487-1493.
43. Sasaki Y, Honmou O: [Bone marrow stem cell therapy for stroke]. Nihon Rinsho 2011, 69(12):2203-2208.
44. van Velthoven CT, Sheldon RA, Kavelaars A, Derugin N, Vexler ZS, Willemen HL, Maas M, Heijnen CJ, Ferriero DM: Mesenchymal stem cell transplantation attenuates brain injury after neonatal stroke. Stroke; a journal of cerebral circulation 2013, 44(5):1426-1432.
45. Shyu WC, Lin SZ, Chiang MF, Su CY, Li H: Intracerebral peripheral blood stem cell (CD34+) implantation induces neuroplasticity by enhancing beta1 integrin-mediated angiogenesis in chronic stroke rats. The Journal of neuroscience : the official journal of the Society for Neuroscience 2006, 26(13):3444-3453.
investigation 2008, 118(7):2482-2495.
47. Tada Y: [Motor association cortex activity in Parkinson's disease--a functional MRI study]. Rinsho Shinkeigaku 1998, 38(8):729-735.
48. Bartzokis G, Cummings JL, Markham CH, Marmarelis PZ, Treciokas LJ, Tishler TA, Marder SR, Mintz J: MRI evaluation of brain iron in earlier- and later-onset Parkinson's disease and normal subjects. Magn Reson Imaging 1999, 17(2):213-222.
49. Hutchinson M, Raff U: Parkinson's disease: a novel MRI method for determining structural changes in the substantia nigra. Journal of neurology, neurosurgery, and psychiatry 1999, 67(6):815-818.
50. Steiner I, Gomori JM, Melamed E: Features of brain atrophy in
Parkinson's disease. A CT scan study. Neuroradiology 1985, 27(2):158-160.
51. Recchia A, Debetto P, Negro A, Guidolin D, Skaper SD, Giusti P:
Alpha-synuclein and Parkinson's disease. FASEB journal : official
publication of the Federation of American Societies for Experimental Biology 2004, 18(6):617-626.
52. Gispert S: Transgenic mice expressing mutant A53T human
alpha-synuclein show neuronal dysfunction in the absence of aggregate formation. Mol Cell Neurosci 2003, 24(2):419-429.
53. Hashimoto M, Rockenstein E, Masliah E: Transgenic models of
alpha-synuclein pathology: past, present, and future. Ann N Y Acad Sci 2003, 991:171-188.
54. Agrawal AK, Shukla S, Chaturvedi RK, Seth K, Srivastava N, Ahmad A, Seth PK: Olfactory ensheathing cell transplantation restores functional deficits in rat model of Parkinson's disease: a cotransplantation approach with fetal ventral mesencephalic cells. Neurobiol Dis 2004, 16(3):516-526.
55. Murrell W, Wetzig A, Donnellan M, Feron F, Burne T, Meedeniya A, Kesby J, Bianco J, Perry C, Silburn P et al: Olfactory mucosa is a potential source for autologous stem cell therapy for Parkinson's disease. Stem Cells 2008, 26(8):2183-2192.
56. Tharion G, Indirani K, Durai M, Meenakshi M, Devasahayam SR, Prabhav NR, Solomons C, Bhattacharji S: Motor recovery following olfactory ensheathing cell transplantation in rats with spinal cord injury. Neurol India 2011, 59(4):566-572.
57. Wu A, Lauschke JL, Gorrie CA, Cameron N, Hayward I, Mackay-Sim A,
Waite PM: Delayed olfactory ensheathing cell transplants reduce nociception after dorsal root injury. Experimental neurology 2011, 229(1):143-157.
捌、圖表 Figure 1
(A)
(B)
DAPI
DAPI
DAPI
DAPI Tuj1
Tuj1 Nestin
Nestin Merge
Merge
Merge
Merge
100X
100X 400X
400X
DAPI Tuj1 Merge
DAPI Tuj1 Merge
(C)
(D)
Figure 1. The timeline for iPS-OSH differentiation and morphology.
(A) The differentiation process and the changes in appearance. Cell changed its morphology from clone to neuron like. (B) Data of immunofluorescence express high level of neural stem cells marker (Nestin and Tuj1). (C) The Stem cells specificity marker (Oct-4) reduced during differentiation stage, and the neural specific marker (Pax6) increased during differentiation stage. (D) There was 63.2% neural stem cells (p75+) in all population by flow analysis.
Oct-4
Pax 6
Actin
iPS-OSH EB NSC
Figure 2
(A) (B)
(C)
Figure 2. Location of surgery and the damage for the affected area.
(A) Tied of right common carotid artery to blocking blood flow. (B) Tied of right middle cerebral artery to blocking blood flow. (C) The damage level after surgery by TTC staining, the white performs showed the area of ischemic stroke.
Figure 3
(A) (B)
Figure 3. Behavioral test data of spontaneous behavior and coordinate behavior.
(A) Locomotor activate was tested the mice spontaneous behavior. (B) Rptarod was tested the mice coordinate behavior. Both locomotor activate and rotarod improved at day 7 and day 14 after iPS-OSH derived NSC transplanted.
Figure 4
(A) (B)
Figure 4. Behavioral test data of poised behavior.
(A) The time for mice through the beam. (B) The numbers of mice foot slipping for mice through the beam. Both time and numbers of slipping have reduced at day 14 and day 21 after iPS-OSH derived NSC transplanted.
Figure 5
(A) (B)
Figure 5. The exterior of non-therapeutic group and therapeutic group.
(A) Non-therapeutic group, the brain was ulceration. (B) Therapeutic group, the brain had little discoloration without ulceration.
↑
↑
↑
↑
↑
↑
↑
↑
↑
↑
↑
↑
Figure 6 (A)
(B)
(C)
Figure 6. The differentiate expression of adventitious cells in vivo.
(A) Co-located with Bisbenzimide+ and GFAP+, NSC differentiated into glia. (B) Co-located with Bisbenzimide+ and NeuN+, NSC differentiated into differentiating neurons. (C) Co-located with Bisbenzimide+ and Tuj1+, NSC differentiated into the kind of cells below to CNS.
Figure 7 (A)
(B)
(C)
Figure 7. Behavioral test data used to evaluate the level of induced Parkinson disease and recover level.
(A) The time for mice through the beam. (B) The numbers of mice foot slipping for mice through the beam. (C) Rptarod was tested the mice coordinate behavior. All data showed that, after MPTP injected, mice behavior decrease. And gradually reply in therapy group (cells、BP、
cells+BP) at day 5, especially in cells+BP group.
Figure 8
(A) (B)
(C) (D)
(E) (F)
Figure 8. Changing in the number of TH cells in the substantia nigra at day 23.
(A) TH expressed in solvent control group at day 23. (B) TH expressed in MPTP group at day 23. (C) TH expressed in cells treatment group at day 23. (D) TH expressed in cells and BP co-treatment group at day 23. (E) TH expressed in BP treatment group at day 23. (F) Quantify of the TH+ cells. Number of TH cells in the therapy group (cells、BP、cells+BP) had increase at day 23, especially in cells+BP group.
Figure 9
(A) (B)
(C) (D)
(E) (F)
Figure 9. The colocalized of the iPS-OSH derived NSC and the differentiation marker.
(A) Co-located with Bisbenzimide+ and TH+, NSC differentiated into dopaminergic neurons in iPS-OSH derived NSC treatment group. (B) Co-located with Bisbenzimide+ and GFAP+, NSC differentiated into glia cells in iPS-OSH derived NSC treatment group. (C) Co-located with Bisbenzimide+ and NeuN+, NSC differentiated into differentiating neurons in iPS-OSH derived NSC treatment group. (D) Co-located with Bisbenzimide+ and TH+, NSC differentiated into dopaminergic neurons in iPS-OSH derived NSC and BP co-treatment group. (E) Co-located with Bisbenzimide+ and GFAP+, NSC differentiated into glia cells in iPS-OSH derived NSC and BP co-treatment group. (F) Co-located with Bisbenzimide+ and NeuN+, NSC differentiated into differentiating neurons in iPS-OSH derived NSC and BP co-treatment group.
Figure 10
(A) (B)
(C) (D)
(E) (F)
TH+ Cell Quanify
0 1 2
3 Solvent Control
MPTP MPTP+Cell MPTP+BP MPTP+Cell+BP
*
*
TH
Figure 10. Recovered of dopaminergic neurons in substantia nigra on day 5.
(A) TH expressed in solvent control group at day 5. (B) TH expressed in MPTP group at day 5. (C) TH expressed in cell treatment group at day 5.
(D) TH expressed in cells and BP co-treatment group at day 5. (E) TH expressed in BP treatment group at day 5. The data showed that TH cells had been signs of reply in therapy group (cells、BP、cells+BP) at day 5.
Figure 11
(A) (B)
(C) (D)
(E) (F)
TH+ Cell Quanify
0 20 40
60 Solvent Control
MPTP MPTP+Cell MPTP+BP MPTP+Cell+BP
**
TH
Figure 11. DA expressed level in striatum on day 5.
(A) TH expressed in solvent control group at day 5. (B) TH expressed in MPTP group at day 5. (C) TH expressed in cell treatment group at day 5.
(D) TH expressed in cells and BP co-treatment group at day 5. (E) TH expressed in BP treatment group at day 5. The data showed that TH cells had been signs of reply in therapy group (cells、BP、cells+BP) at day 5.
Figure 12
(A) (B)
(C) (D)
(E) (F)
Iba1+ Cell Quanify
0.0 0.5 1.0
1.5 Solvent Control
MPTP MPTP+Cell MPTP+BP MPTP+Cell+BP
*** *** *** ***
Iba1
Figure 12. Performance of brain injury in striatum on day 5.
(A) Iba1 expressed in solvent control group at day 5. (B) Iba1 expressed in MPTP group at day 5. (C) Iba1 expressed in cell treatment group at day 5. (D) Iba1 expressed in cells and BP co-treatment group at day 5. (E) Iba1 expressed in BP treatment group at day 5. The data showed that express of Iba1 had been signs of reduce in therapy group (cells、BP、
cells+BP) at day 5.
Figure 13
(A) (B)
(C) (D)
(E) (F)
KI67+ Cell Quanify
0 10 20 30
40 Solvent Control
MPTP MPTP+Cell MPTP+BP MPTP+Cell+BP
KI67
Figure 13. Performance of cell proliferated in striatum on day 5.
(A) KI67 expressed in solvent control group at day 5. (B) KI67 expressed in MPTP group at day 5. (C) KI67 expressed in cells treatment group at day 5. (D) KI67 expressed in cells and BP co-treatment group at day 5. (E) KI67 expressed in BP treatment group at day 5. The data showed that after MPTP inject, it couldn’t cells proliferate. After treatment, reply cell proliferate.
Figure 14
Figure 14. ELISA for detecting dopamine (DA) concentration in serum.
The concentration of DA was reduced in MPTP group at day 9, but the therapy group (cells、BP、cells+BP) had reply to closed the normal level.
At day 23, DA reply to closed the normal level, and the therapy group (cells、BP、cells+BP) maintain the level.
ELISA
Day 9 Day 23
0 1 2 3
4 Solvent Control
MPTP MPTP+Cell MPTP+BP MPTP+Cell+BP
*** ***
*** ***
***
*** * ***
Log of concentraction
Figure 15
(A) (B)
(C) (D)
(E) (F)
SC MPTP
Cell Cell+BP
BP TUNEL
0.0 0.5 1.0
1.5 Solvent Control
MPTP MPTP+Cell MPTP+BP MPTP+Cell+BP
**
**
**
*
Figure 15. TUNEL for the cell therapy mechanism.
(A) TUNEL analysis in solvent control group at day 5. (B) TUNEL analysis in MPTP group at day 5. (C) TUNEL analysis in cells treatment group at day 5. (D) TUNEL analysis in cells and BP co-treatment group at day 5. (E) TUNEL analysis in BP group at day 5. (F) Quantitative data.
TUNEL showed that after therapy, a significant reduction in apoptosis.
Table 1
http://www.doh.gov.tw/cht2006/index_populace.aspx
Table 1. Raining cause of death in Taiwan
A00-Y98 153,823 661.0 450.6 100.0 152,030 655.5 462.4 100.0 0.8 5.5 -11.8
-C00-C97 1 43,665 187.6 131.3 28.4 1 42,559 183.5 132.2 28.0 2.3 4.1 -0.9 0.4
I01-I02.0, I05-I09,
I20-I25, I27, I30-I52 2 17,121 73.6 47.9 11.1 2 16,513 71.2 47.9 10.9 3.3 2.4 -0.0 0.3
I60-I69 3 11,061 47.5 30.8 7.2 3 10,823 46.7 31.3 7.1 1.9 0.9 -0.5 0.1
J12-J18 4 9,314 40.0 24.4 6.1 5 9,047 39.0 24.8 6.0 2.6 1.0 -0.4 0.1
E10-E14 5 9,281 39.9 26.5 6.0 4 9,081 39.2 26.9 6.0 1.9 0.7 -0.4 0.1
V01-X59, Y85-Y86 6 6,873 29.5 23.8 4.5 6 6,726 29.0 24.1 4.4 1.8 0.5 -0.3 0.0
J40-J47 7 6,326 27.2 16.4 4.1 7 5,984 25.8 16.2 3.9 5.4 1.4 0.2 0.2
I10-I15 8 4,986 21.4 13.3 3.2 9 4,631 20.0 12.9 3.0 7.3 1.5 0.4 0.2
K70, K73-K74 9 4,975 21.4 15.6 3.2 8 5,153 22.2 16.5 3.4 -3.8 -0.8 -0.9 -0.2
N00-N07, N17-N19,
N25-N27 10 4,327 18.6 12.1 2.8 10 4,368 18.8 12.6 2.9 -1.3 -0.2 -0.5 -0.1
35,894 154.2 108.6 23.3 37,145 160.2 116.8 24.4 -3.7 -5.9 -8.3 -1.1
X60-X84, Y87.0 11 3,766 16.2 13.1 2.4 12 3,507 15.1 12.3 2.3 7.0 1.1 0.8 0.1
A40-A41 12 3,671 15.8 10.0 2.4 11 3,886 16.8 11.0 2.6 -5.8 -1.0 -1.0 -0.2
R54 13 1,359 5.8 3.2 0.9 13 1,504 6.5 3.8 1.0 -9.9 -0.6 -0.6 -0.1
M00-M99 14 1,244 5.3 3.7 0.8 14 1,280 5.5 3.9 0.8 -3.1 -0.2 -0.2 -0.0
D00-D48 15 1,091 4.7 3.1 0.7 15 1,067 4.6 3.2 0.7 1.9 0.1 -0.0 0.0
Table 2
北醫藥訊 2007.36:7-11
Table 2. Drug treatment for Parkinson