紙型免疫吸附法(Paper-based ELISA)於2008年開始被發展出來，其用濾紙來做為中心的吸水區是直徑約1公分的圓形，而用噴蠟的印表機製作出環繞此圓形的疏水區，而後再用烤盤將蠟從最上層溶解至濾紙最下層，即完成試片的基本製作，一個測試區僅需2~3微升(microliter) 的檢體量即可完成一次測試。另一方面，在臨床眼科上，前房液中的血管新生因子(Vascular endothelial growth factor, VEGF)濃度已被大量證實在兩大導致失明主要的疾病：糖尿病視網膜病變及老年性黃斑部病變中扮演重要的致病角色，據估計每年全球有施行數十萬次以上的抗血管新生因子抗體注射手術。然而臨床眼科實務上，前房液每次採集僅能提供100至200微升(microliter)的檢體體積，使得傳統的生化定量血管新生因子濃度的方法變得很困難，也因此檢體上的匱乏，本研究中即利用紙型免疫吸附法每次僅需2 microliter的特性，來運用在前房液血管新生因子濃度的定量上。本論文內容闡述研究的四個大部分，第一個部分是闡述開發平台：紙型ELISA(paper-based ELISA)在眼科已經確診的病人中採集前房液檢體(檢體數目=54)後，去量測不同視網膜缺氧疾病下其血管新生因子(Vascula endothelial growth factor, VEGF)濃度，在不同疾病下的其血管新生因子濃度已有顯著差異，第二部分是探討臨床上前房液VEGF濃度在眼科疾病中更細微精確的角色及可運用的發展，眼睛中血管新生因子扮演如同Biomarker，在疾病進展的過程中，是VEGF先上升，而後視網膜結構產生惡化(例如水腫、出血等)，最後是不樂見的功能被破壞:視力損傷；第三部分是使用紙型ELISA在追蹤27個病人(檢體數目=46)在接受抗血管新生抗體治療前與治療後的前房液VEGF濃度，紙型ELISA已成功半定量治療前後的濃度亦達到顯著差異，在追蹤上也看出VEGF上升的趨勢因抗體不同而有不同趨勢；最後第四部分是紙型ELISA可以再優化、減少誤差及未來可能的應用與發展。

Paper-based ELISA was developed around 2008. It was cellulose-based platform that using wax printing to form hydrophobic zone and cellulose to form the central portion as hydrophilic zone. The colorimetric reaction can be accomplished within a 1 cm-diameter hydrophilic zone. It took only 2~3 μL to accomplish one test in one test zone. On the other hand, in clinical ophthalmology, VEGF level in aqueous humor was proved to play an important role in the pathogenesis of diabetic retinopathy and age-related macular degeneration. It’s estimated that >500,000 times of anti-VEGF injections were performed around the world. However, scarcity of aqueous humor limits the development of biochemical method to qualifying or quantifying the existence of biomarker like VEGF through aqueous humor because only 100~200 μL can be acquired through aqueous humor per sampling. Thus, we choose paper-based ELISA to quantifying aqueous humor VEGF level. This PhD paper consisted of four parts: (1) Establishing protocol of P-ELISA for aqueous VEGF. Significant difference between different disease spectrum were proved. (2) Identify the rationale for aqueous VEGF as a biomarker and elevated first. Then retinal structure changes happened later. Finally, ocular function decline as vision impair happened last. (3) P-ELISA successfully determine aqueous VEGF level before and after treatment. (4) Further work: optimization and possible application for P-ELISA in ophthalmology.

Chapter 1. Introduction of Paper-Based ELISA 7
Chapter 2. Monitoring the VEGF Level in Aqueous Humor of Patients with Ophthalmologically relevant Diseases via Ultrahigh Sensitive Paper-based ELISA (以超高靈敏度紙型免疫吸附法監測眼科相關疾病患者之前房液中血管內皮生長因子之濃度)...9
2.1 Introduction 實驗簡介.....................9
2.2 Material and Methods:材料與方法…..............................................12
2.2.1 Antibodies and antigens抗體與抗原 .........12
2.2.2 Aqueous humor from patients採集患者之前房液.................13
2.2.3 Color fundus photographs and fluorescein angiography 彩色眼底攝影與螢光血管攝影..........13
2.2.4 P-ELISA for detection of VEGF level from patients以P-ELISA 偵測患者之VEGF濃度......................13
2.2.5 Quantifying the intensity of test zone through a scanner 以掃描器定量測試區的訊號強度.................14
2.3 Results and discussion結果與討論......... 15
2.3.1 Characteristics and calibration of paper-based VEGF test紙型VEGF測試的特性與校準.................15
2.3.2 Device performance with retinal ischemic condition clinical samples視網膜缺血疾病之臨床檢體偵測效能 ................................................20
2.3.3 Discussions about P-ELISA for aqueous VEGF detection P-ELISA偵測前房液VEGF之討論........22
2.4 Conclusion結論...........................24

Chapter 3. Monitoring VEGF Level with Low-Volume Sampling in Major Vision-threatening Diseases: Age-related Macular Degeneration and Diabetic Retinopathy. 監測威脅視力的重大疾病之低量檢體中VEGF濃度：老年性黃斑部退化與糖尿病視網膜病變...25
3.1 Introduction 實驗簡介..........................25
3.1.1 Critical role of VEGF in the pathogenesis of two major vision-threatening diseases: age-related macular degeneration and diabetic retinopathy( VEGF於兩大視力威脅疾病之致病機轉的關鍵作用：老年性黃斑部病變與糖尿病視網膜病變)..25
3.1.2 Available optical imaging modalities for evaluation of treatment response: fluorescence angiography (FAG) and optical coherence tomography (OCT) 光學影像造影評估治療反應：螢光血管攝影(FAG)及光學同調斷層掃描(OCT).................................................27
3.1.3 目前可得的ELISA 生化方法評估VEGF: ELISA and Luminex bead-based(微珠式) ELISA ......................28
3.2 New innovations in microliter-scaled sample diagnostics 診斷微升體積檢體的創新方式........29
3.2.1. Paper-based ELISA 紙型ELISA..................30
3.2.2. Further POC diagnostic devices that could provide ophthalmologists with instant results to aid in re-treatment determination進一步提供眼科醫師即時結果，以幫助臨床醫師決定再治療的POC診斷設備..........................33
3.3 Importance of surveillance for VEGF detection 使用VEGF偵測來監控的重要性 ..............................33
3.3.1 P-ELISA as an easy-to-use tool for surveillance of ocular VEGF levels: P-ELISA 為容易操作的眼部VEGF濃度監測工具....................................35
3.4 Conclusion結論.................................37
Chapter 4. Paper-based ELISA detecting aqueous VEGF level in follow-up scenarios. 紙型ELISA成功的檢測出前房液VEGF濃度在接受抗血管新生因子抗體治療的病人族群.............................................38
4.1 Introduction實驗簡介 ..............................38
4.2 Material and Methods:材料與方法...................39
4.2.1 Subjects 受試者...............39
4.2.2 Aqueous Humor Collection and IVI Injection 前房液採集與玻璃體內注射......................40
4.2.3 Measurement of Vascular Endothelial Growth Factor 測量血管內皮生長因子...................40
4.2.4 Measurement of central foveal thickness and visual acuity測量中央視網膜厚度與視力..........41
4.2.5 Statistical Analysis 統計分析.............41
4.3 Results: 實驗結果...........................42
4.3.1 Basic Characteristics基本特性..........42
4.3.2 Vascular endothelial growth factor (VEGF) concentration before and after IVI , 玻璃體注射前後之血管內皮生長因子(VEGF)濃度.....................44
4.3.3 CFT and visual acuity before and after IVI 玻璃體注射前後之中央黃斑部厚度和視力.............48
4.4 Discussion 實驗討論....................49
4.5 Conclusion 結論........................52

Chapter 5. Future work and possible applications.....53
5.1 Introduction簡介..........53
5.2 優化紙型免疫吸附法誤差及精準度..................53
5.3 Monitor aqueous VEGF level in patients with DM 監測糖尿病患者前房液中的血管新生因子濃度 .............54
5.4 Monitoring VEGF level for patient with Age-related Macular Degeneration 監測老年性黃斑部病變患者前房液的血管新生因子濃度.......56
5.5 Animal model for PDR and AMD建立糖尿病視網膜病變及 老年性黃斑部病變動物模式.......57
5.6 Paper-Based ELISA for aqueous cytokine detection in patients with uveitis or other inflammatory disease. 利用紙型免疫吸附法的低檢體量特性來檢測眼球發炎反應(如虹彩炎等)前房液中的細胞激素.........58
5.7 Safer Micro-Needle for aqueous humor acquisition 開發更安全的微針吸取來使得前房抽取術更有可得與運用性.....59
5.8總結...................... 60
Reference..............61

(1) Cheng, C. M.; Martinez, A. W.; Gong, J.; Mace, C. R.; Phillips, S. T.; Carrilho, E.; Mirica, K. A.; Whitesides, G. M. Paper-based ELISA. Angew Chem Int Ed Engl 2010, 49, 4771-4774.
(2) Wang, H. K.; Tsai, C. H.; Chen, K. H.; Tang, C. T.; Leou, J. S.; Li, P. C.; Tang, Y. L.; Hsieh, H. J.; Wu, H. C.; Cheng, C. M. Cellulose-Based Diagnostic Devices for Diagnosing Serotype-2 Dengue Fever in Human Serum. Advanced healthcare materials 2013.
(3) Hsu, C. K.; Huang, H. Y.; Chen, W. R.; Nishie, W.; Ujiie, H.; Natsuga, K.; Fan, S. T.; Wang, H. K.; Lee, J. Y.; Tsai, W. L.; Shimizu, H.; Cheng, C. M. Paper-based ELISA for the detection of autoimmune antibodies in body fluid-the case of bullous pemphigoid. Anal Chem 2014, 86, 4605-4610.
(4) Yamada, K.; Takaki, S.; Komuro, N.; Suzuki, K.; Citterio, D. An antibody-free microfluidic paper-based analytical device for the determination of tear fluid lactoferrin by fluorescence sensitization of Tb3+. The Analyst 2014, 139, 1637-1643.
(5) Gubala V., Leanne F. Harris LF, Ricco AJ, Tan MX, and Williams DE. Point of Care Diagnostics: Status and Future. Anal. Chem., 2012, 84 (2), pp 487–515
(6) Yager P, Domingo GJ, Gerdes J. Point-of-Care Diagnostics for Global Health. Annu Rev Biomed Eng. 2008;10:107-44.
(7) Mansfield E, O'Leary TJ, Gutman SI. Food and Drug Administration regulation of in vitro diagnostic devices. J Mol Diagn. 2005 Feb;7(1):2-7.
(8) Adamis AP, M. J., Bernal MT, D'Amico DJ, Folkman J, Yeo TK, Yeo KT. Increased vascular endothelial growth factor levels in the vitreous of eyes with proliferative diabetic retinopathy. American Journal of Ophthalmology 1994, 118, 6.
(9) Shams, N.; Ianchulev, T. Role of vascular endothelial growth factor in ocular angiogenesis. Ophthalmol Clin North Am 2006, 19, 335-344.
(10) Wong, T. Y.; Scott, I. U. Clinical practice. Retinal-vein occlusion. N Engl J Med 2010, 363, 2135-2144.
(11) Philip J. Rosenfeld, M. D., Ph.D., David M. Brown, M.D., Jeffrey S. Heier, M.D., David S. Boyer, M.D., ; Peter K. Kaiser, M. D., Carol Y. Chung, Ph.D., and Robert Y. Kim, M.D., for the MARINA Study Group. Ranibizumab for Neovascular Age-Related Macular Degeneration. N Engl J Med 2006, 355, 13.
(12) Sawada, O.; Kawamura, H.; Kakinoki, M.; Sawada, T.; Ohji, M. Vascular endothelial growth factor in aqueous humor before and after intravitreal injection of bevacizumab in eyes with diabetic retinopathy. Archives of ophthalmology 2007, 125, 1363-1366.
(13) de Jong, P. T. Age-related macular degeneration. N Engl J Med 2006, 355, 1474-1485.
(14) Funatsu, H.; Yamashita, H.; Noma, H.; Mimura, T.; Nakamura, S.; Sakata, K.; Hori, S. Aqueous humor levels of cytokines are related to vitreous levels and progression of diabetic retinopathy in diabetic patients. Graefe's archive for clinical and experimental ophthalmology = Albrecht von Graefes Archiv fur klinische und experimentelle Ophthalmologie 2005, 243, 3-8.
(15) Campochiaro, P. A.; Choy, D. F.; Do, D. V.; Hafiz, G.; Shah, S. M.; Nguyen, Q. D.; Rubio, R.; Arron, J. R. Monitoring ocular drug therapy by analysis of aqueous samples. Ophthalmology 2009, 116, 2158-2164.
(16) Sharma, R. K.; Rowe-Rendleman, C. L. Validation of molecular and genomic biomarkers of retinal drug efficacy: use of ocular fluid sampling to evaluate VEGF. Neurochemical research 2011, 36, 655-667.
(17) Funatsu, H.; Yamashita, H.; Noma, H.; Mimura, T.; Yamashita, T.; Hori, S. Increased levels of vascular endothelial growth factor and interleukin-6 in the aqueous humor of diabetics with macular edema. American Journal of Ophthalmology 2002, 133, 70-77.
(18) Patel, J. I.; Tombran-Tink, J.; Hykin, P. G.; Gregor, Z. J.; Cree, I. A. Vitreous and aqueous concentrations of proangiogenic, antiangiogenic factors and other cytokines in diabetic retinopathy patients with macular edema: Implications for structural differences in macular profiles. Experimental eye research 2006, 82, 798-806.
(19) Rajesh K. Sharma, A. T. R., K.V. Chalam. Multiplex immunoassay analysis of biomarkers in clinically accessible quantities of human aqueous humor. Molecular Vision 2009, 15, 10.
(20) Mitchell, P.; Korobelnik, J. F.; Lanzetta, P.; Holz, F. G.; Prunte, C.; Schmidt-Erfurth, U.; Tano, Y.; Wolf, S. Ranibizumab (Lucentis) in neovascular age-related macular degeneration: evidence from clinical trials. Br J Ophthalmol 2010, 94, 2-13.
(21) Lo, S. J.; Yang, S. C.; Yao, D. J.; Chen, J. H.; Tu, W. C.; Cheng, C. M. Molecular-level dengue fever diagnostic devices made out of paper. Lab on a chip 2013, 13, 2686-2692.
(22) Pollock, N. R.; Rolland, J. P.; Kumar, S.; Beattie, P. D.; Jain, S.; Noubary, F.; Wong, V. L.; Pohlmann, R. A.; Ryan, U. S.; Whitesides, G. M. A paper-based multiplexed transaminase test for low-cost, point-of-care liver function testing. Science translational medicine 2012, 4, 152ra129.
(23) Wu, H.-Y.; Rubinstein, M.; Shih, E.; Guttag, J.; Durand, F.; Freeman, W. Eulerian video magnification for revealing subtle changes in the world. ACM Transactions on Graphics 2012, 31, 1-8.
(24) F Aouizerate, J. C., L Poirier, Ph Verin, A Cheyrou, J Begueret, F Lagoutte. Detection of Toxoplasma gondii in aqueous humour
by the polymerase chain reaction. Br J Ophthalmol 1993, 77, 3.
(25) Germán Bou, M. S. F., Paloma Martí-Belda, Enrique Navas and Antonio Guerrero. Value of PCR for Detection of Toxoplasma gondii
in Aqueous Humor and Blood Samples from Immunocompetent Patients with Ocular Toxoplasmosis. J. Clin. Microbiol. 1999, 37, 4.
(26) Tolentino, M. J. Current Molecular Understanding and Future Treatment Strategies for Pathologic Ocular Neovascularization. Curr Mol Med 2009, 9, 973-981.
(27) Stewart, M. W.; Rosenfeld, P. J.; Penha, F. M.; Wang, F.; Yehoshua, Z.; Bueno-Lopez, E.; Lopez, P. F. Pharmacokinetic rationale for dosing every 2 weeks versus 4 weeks with intravitreal ranibizumab, bevacizumab, and aflibercept (vascular endothelial growth factor Trap-eye). Retina 2012, 32, 434-457.
(28) Krohne, T. U.; Eter, N.; Holz, F. G.; Meyer, C. H. Intraocular pharmacokinetics of bevacizumab after a single intravitreal injection in humans. Am J Ophthalmol 2008, 146, 508-512.
(29) Sankarathi Balaiya, S. g., Ravi K Murthy, Kakarla V Chalam. Freezing adversely affects measurement of vascular endothelial growth factor levels in human aqueous samples. Clinical Ophthalmology 2011, 5, 5.
(30) James C. Folk, M. D., and Edwin M. Stone, M.D., Ph.D. Ranibizumab Therapy for Neovascular Age-Related Macular Degeneration. New Engl J Med 2010, 363, 8.
(31) Mitchell, P. A systematic review of the efficacy and safety outcomes of anti-VEGF agents used for treating neovascular age-related macular degeneration: comparison of ranibizumab and bevacizumab. Current medical research and opinion 2011, 27, 1465-1475.
(32) Campbell, R. J.; Bell, C. M.; Campbell Ede, L.; Gill, S. S. Systemic effects of intravitreal vascular endothelial growth factor inhibitors. Curr Opin Ophthalmol 2013, 24, 197-204.
(33) Zakaria, N.; Van Grasdorff, S.; Wouters, K.; Rozema, J.; Koppen, C.; Lion, E.; Cools, N.; Berneman, Z.; Tassignon, M. J. Human tears reveal insights into corneal neovascularization. PloS one 2012, 7, e36451.
(34) Vu, H. T.; Keeffe, J. E.; McCarty, C. A.; Taylor, H. R. Impact of unilateral and bilateral vision loss on quality of life. Br J Ophthalmol 2005, 89, 360-363.
(35) Cheung, N.; Mitchell, P. ; Wong, T. Y. Diabetic retinopathy. The Lancet 2010, 376, 124-136.
(36) Lim, L. S.; Mitchell, P.; Seddon, J. M.; Holz, F. G.; Wong, T. Y. Age-related macular degeneration. The Lancet 2012, 379, 1728-1738.
(37) Jager, R. D.; Mieler, W. F.; Miller, J. W. Age-related macular degeneration. N Engl J Med 2008, 358, 2606-2617.
(38) Engelgau, M. M.; Geiss, L. S.; Saaddine, J. B.; Boyle, J. P.; Benjamin, S. M.; Gregg, E. W.; Tierney, E. F.; Rios-Burrows, N.; Mokdad, A. H.; Ford, E. S.; Imperatore, G.; Narayan, K. M. The evolving diabetes burden in the United States. Annals of internal medicine 2004, 140, 945-950.
(39) Penn, J. S.; Madan, A.; Caldwell, R. B.; Bartoli, M.; Caldwell, R. W.; Hartnett, M. E. Vascular endothelial growth factor in eye disease. Progress in retinal and eye research 2008, 27, 331-371.
(40) Campochiaro, P. A.; Hafiz, G.; Shah, S. M.; Nguyen, Q. D.; Ying, H.; Do, D. V.; Quinlan, E.; Zimmer-Galler, I.; Haller, J. A.; Solomon, S. D.; Sung, J. U.; Hadi, Y.; Janjua, K. A.; Jawed, N.; Choy, D. F.; Arron, J. R. Ranibizumab for macular edema due to retinal vein occlusions: implication of VEGF as a critical stimulator. Molecular therapy : the journal of the American Society of Gene Therapy 2008, 16, 791-799.
(41) Silverman, E. Setting the Bar Higher For Off-Label Use of Biologics. Biotechnol Healthc 2011, 8, 14-18.
(42) dell'Omo, R.; Cassetta, M.; dell'Omo, E.; di Salvatore, A.; Hughes, J. M.; Aceto, F.; Porcellini, A.; Costagliola, C. Aqueous humor levels of vascular endothelial growth factor before and after intravitreal bevacizumab in type 3 versus type 1 and 2 neovascularization. A prospective, case-control study. Am J Ophthalmol 2012, 153, 155-161 e152.
(43) Muether, P. S.; Hermann, M. M.; Viebahn, U.; Kirchhof, B.; Fauser, S. Vascular endothelial growth factor in patients with exudative age-related macular degeneration treated with ranibizumab. Ophthalmology 2012, 119, 2082-2086.
(44) Kuzmin, A.; Lipatov, D.; Chistyakov, T.; Smirnova, O.; Arbuzova, M.; Ilin, A.; Shestakova, M.; Dedov, I. Vascular endothelial growth factor in anterior chamber liquid patients with diabetic retinopathy, cataract and neovascular glaucoma. Ophthalmology and therapy 2013, 2, 41-51.
(45) Kumar, S.; Kumar, S.; Ali, M. A.; Anand, P.; Agrawal, V. V.; John, R.; Maji, S.; Malhotra, B. D. Microfluidic-integrated biosensors: prospects for point-of-care diagnostics. Biotechnology journal 2013, 8, 1267-1279.
(46) Nguyen, Q. D.; Brown, D. M.; Marcus, D. M.; Boyer, D. S.; Patel, S.; Feiner, L.; Gibson, A.; Sy, J.; Rundle, A. C.; Hopkins, J. J.; Rubio, R. G.; Ehrlich, J. S.; Rise; Group, R. R. Ranibizumab for diabetic macular edema: results from 2 phase III randomized trials: RISE and RIDE. Ophthalmology 2012, 119, 789-801.
(47) Schein, O. D.; Tielsch, J. M.; Muñoz, B.; Bandeen-Roche, K.; West, S. Relation between Signs and Symptoms of Dry Eye in the Elderly. Ophthalmology 1997, 104, 1395-1401.
(48) Andres W. Martinez, S. T. P., Emanuel Carrilho, Samuel W. Thomas III, Hayat Sindi, and George M. Whitesides. Simple Telemedicine for Developing Regions Camera Phones and Paper Based Microfluidic Devices for Real Time Off Site Diagnosis.pdf. Analytical Chemistry 2008, 80, 9.
(49) Yetisen, A. K.; Akram, M. S.; Lowe, C. R. Paper-based microfluidic point-of-care diagnostic devices. Lab on a chip 2013, 13, 2210-2251.
(50) Jani, I. V.; Peter, T. F. How point-of-care testing could drive innovation in global health. N Engl J Med 2013, 368, 2319-2324.
(51) Hsu, M. Y.; Yang, C. Y.; Hsu, W. H.; Lin, K. H.; Wang, C. Y.; Shen, Y. C.; Chen, Y. C.; Chau, S. F.; Tsai, H. Y.; Cheng, C. M. Monitoring the VEGF level in aqueous humor of patients with ophthalmologically relevant diseases via ultrahigh sensitive paper-based ELISA. Biomaterials 2014, 35, 3729-3735.
(52) Selim, K. M.; Sahan, D.; Muhittin, T.; Osman, C.; Mustafa, O. Increased levels of vascular endothelial growth factor in the aqueous humor of patients with diabetic retinopathy. Indian journal of ophthalmology 2010, 58, 375-379.
(53) Pai, N. P.; Vadnais, C.; Denkinger, C.; Engel, N.; Pai, M. Point-of-care testing for infectious diseases: diversity, complexity, and barriers in low- and middle-income countries. PLoS medicine 2012, 9, e1001306.
(54) Kwan, A. S.; Barry, C.; McAllister, I. L.; Constable, I. Fluorescein angiography and adverse drug reactions revisited: the Lions Eye experience. Clinical & experimental ophthalmology 2006, 34, 33-38.
(55) Yang, C.-S.; Sung, C.-S.; Lee, F.-L.; Hsu, W.-M. Management of Anaphylactic Shock During Intravenous Fluorescein Angiography at an Outpatient Clinic. Journal of the Chinese Medical Association 2007, 70, 348-349.
(56) Falavarjani, K. G.; Nguyen, Q. D. Adverse events and complications associated with intravitreal injection of anti-VEGF agents: a review of literature. Eye 2013, 27, 787-794.
(57) Moja, L.; Lucenteforte, E.; Kwag, K. H.; Bertele, V.; Campomori, A.; Chakravarthy, U.; D'Amico, R.; Dickersin, K.; Kodjikian, L.; Lindsley, K.; Loke, Y.; Maguire, M.; Martin, D. F.; Mugelli, A.; Muhlbauer, B.; Puntmann, I.; Reeves, B.; Rogers, C.; Schmucker, C.; Subramanian, M. L.; Virgili, G. Systemic safety of bevacizumab versus ranibizumab for neovascular age-related macular degeneration. The Cochrane database of systematic reviews 2014, 9, CD011230.
(58) Brown, D. M.; Nguyen, Q. D.; Marcus, D. M.; Boyer, D. S.; Patel, S.; Feiner, L.; Schlottmann, P. G.; Rundle, A. C.; Zhang, J.; Rubio, R. G.; Adamis, A. P.; Ehrlich, J. S.; Hopkins, J. J.; Ride; Group, R. R. Long-term outcomes of ranibizumab therapy for diabetic macular edema: the 36-month results from two phase III trials: RISE and RIDE. Ophthalmology 2013, 120, 2013-2022.
(59) Bressler, N. M. Age-Related Macular Degeneration Is the Leading Cause of Blindness. Jama 2004, 291, 1900.
(60) Hou, X.; Kumar, A.; Lee, C.; Wang, B.; Arjunan, P.; Dong, L.; Maminishkis, A.; Tang, Z.; Li, Y.; Zhang, F.; Zhang, S. Z.; Wardega, P.; Chakrabarty, S.; Liu, B.; Wu, Z.; Colosi, P.; Fariss, R. N.; Lennartsson, J.; Nussenblatt, R.; Gutkind, J. S.; Cao, Y.; Li, X. PDGF-CC blockade inhibits pathological angiogenesis by acting on multiple cellular and molecular targets. Proceedings of the National Academy of Sciences of the United States of America 2010, 107, 12216-12221.
(61) Rajappa, M.; Saxena, P.; Kaur, J. Ocular Angiogenesis: mechanisms and recent advances in therapy. Adv Clin Chem. . 2010, 50, 103-121.
(62) Group, C. R.; Martin, D. F.; Maguire, M. G.; Ying, G. S.; Grunwald, J. E.; Fine, S. L.; Jaffe, G. J. Ranibizumab and bevacizumab for neovascular age-related macular degeneration. N Engl J Med 2011, 364, 1897-1908.
(63) Wang, X.; Sawada, T.; Sawada, O.; Saishin, Y.; Liu, P.; Ohji, M. Serum and plasma vascular endothelial growth factor concentrations before and after intravitreal injection of aflibercept or ranibizumab for age-related macular degeneration. Am J Ophthalmol 2014, 158, 738-744 e731.
(64) Fauser, S.; Schwabecker, V.; Muether, P. S. Suppression of intraocular vascular endothelial growth factor during aflibercept treatment of age-related macular degeneration. Am J Ophthalmol 2014, 158, 532-536.
(65) Lux, A.; Llacer, H.; Heussen, F. M.; Joussen, A. M. Non-responders to bevacizumab (Avastin) therapy of choroidal neovascular lesions. Br J Ophthalmol 2007, 91, 1318-1322.
(66) Susanne B. Loss of reactivity in intravitreal anti-VEGF therapy: tachyphylaxis or tolerance? Br J Ophthalmol 2012;96:1 1-2
(67) Schmidt-Erfurth U, Eldem B, Guymer R, et al. Efficacy and safety of monthly versus quarterly ranibizumab treatment in neovascular age-related macular degeneration: the EXCITE study. Ophthalmology 2011;118:831-839.
(68) Curnow, S. J.; Falciani, F.; Durrani, O. M.; Cheung, C. M.; Ross, E. J.; Wloka, K.; Rauz, S.; Wallace, G. R.; Salmon, M.; Murray, P. I. Multiplex bead immunoassay analysis of aqueous humor reveals distinct cytokine profiles in uveitis. Invest Ophthalmol Vis Sci 2005, 46, 4251-4259.
(69) Hsu, M. Y.; Chen, S. J.; Chen, K. H.;Hung, Y. C.; Tsai, H. Y.; Cheng, C. M.Monitoring VEGF levels with low-volume sampling in major vision-threatening disease: age-related macualr degeneration and diabetic retinopathy. Lab Chip, 2015,15,2357-2363.
(70) Holz FG, Amoaku W, Donate J, et al. Safety and efficacy of a flexible dosing regimen of ranibizumab in neovascular age-related macular degeneration: the SUSTAIN study. Ophthalmology 2011;118:663-671.
(71) Lalwani GA, Rosenfeld PJ, Fung AE, et al. A variable-dosing regimen with intravitreal ranibizumab for neovascular age-related macular degeneration: year 2 of the PrONTO Study. Am J Ophthalmol 2009;148:43-58 e41.
(72) Oubraham H, Cohen SY, Samimi S, et al. Inject and extend dosing versus dosing as needed: a comparative retrospective study of ranibizumab in exudative age-related macular degeneration. Retina 2011;31:26-30.
(73) Bakri SJ, Snyder MR, Reid JM, et al. Pharmacokinetics of intravitreal ranibizumab (Lucentis). Ophthalmology 2007;114:2179-82.
(74)Bergers G, Hanahan D. Modes of resistance to anti-angiogenic therapy. Nature reviews Cancer 2008;8:592-603.
(75)Jo N, Mailhos C, Ju M, et al. Inhibition of platelet-derived growth factor B signaling enhances the efficacy of anti-vascular endothelial growth factor therapy in multiple models of ocular neovascularization. The American journal of pathology 2006;168:2036-2053.
(76)Singer M. Advances in the management of macular degeneration. F1000prime reports 2014;6:29.
(77) Christodouleas DC, Nemiroski A, Kumar AA, Whitesides GM. Broadly Available Imaging Devices Enable High-Quality Low-Cost Photometry. Anal. Chem. 2015, 87, 9170−9178
(78) Piotr Wargocki, Wei Deng , Ayad G. Anwer ; Ewa M. Goldys. Medically Relevant Assays with a Simple Smartphone and Tablet Based Fluorescence Detection System. Sensors 2015, 5, 11653-11664
(79) IDF Diabetes Atlas. 5th Edition:http://www.idf.org/diabetesatlas/5e/the-global-burden
(80) Yau JW, Rogers SL, Kawasaki R,,et al. Global prevalence and major risk factors of diabetic retinopathy. Diabetes Care. 2012 Mar;35(3):556-64
(81) Chen SJ, Cheng CY, Peng KL, Li AF, Hsu WM, Liu JH, Chou P. Prevalence and associated risk factors of age-related macular degeneration in an elderly Chinese population in Taiwan: the Shihpai Eye Study.Invest Ophthalmol Vis Sci. 2008 Jul;49(7):3126-33.
(82) Muether PS, Neuhann I, Buhl C, Hermann MM, Kirchhof B, Fauser S. Intraocular growth factors and cytokines in patients with dry and neovascular age-related macular degeneration. Retina. 2013 Oct;33(9):1809-14
(83) Robinson R., Barathi VA, Chaurasia SS, Wong TY, Kern TS. Update on animal models of diabetic retinopathy: from molecular approaches to mice and higher mammals. Disease Models & Mechanisms 2012,5, 444-456
(84) Shah RS, Soetikno BT, Lajko M, Fawzi AA. A Mouse Model for Laser-induced Choroidal Neovascularization. J Vis Exp. 2015 Dec 27; (106):e53502.
(85) 黃德光 周穎政 蒲正筠 台灣民眾眼睛葡萄膜炎的流行病學研究 國立陽明大學公共衛生研究所 2013
(86) Murray PI, Hoekzema R, van Haren MA, de Hon FD, Kijlstra A. Aqueous humor interleukin-6 levels in uveitis. Invest Ophthalmol Vis Sci. 1990 May;31(5):917-20.
(87) van Kooij B, Rothova A, Rijkers GT, de Groot-Mijnes JD. Distinct Cytokine and Chemokine Profiles in the Aqueous of Patients With Uveitis and Cystoid Macular Edema. Am J Ophthalmol 2006; 142:192–194
(88) Cheung CM, Durrani OM, and Murray PI. The safety of anterior chamber paracentesis in patients with uveitis. Br J Ophthalmol. 2004 Apr; 88(4): 582–583.