癌症一直是全球十大主要死因之一。近年來，癌症有越來越年輕化的趨勢。細胞激素是由細胞分泌的微小蛋白質，可以在癌症發病機制中發揮重要作用。在腫瘤免疫中，腫瘤壞死因子- (TNF-)、介白素-2 (IL-2) 和干擾素- (IFN-) 等細胞激素可以抑制腫瘤細胞生長，而介白素-10 (IL-10) 和介白素-4 (IL-4) 等細胞激素則可促進腫瘤細胞的成長。根據研究，中藥能夠增加抗腫瘤細胞激素的產生，進而改善免疫狀態以對抗癌症的形成和發展。在檢測個人化中草藥實驗中，可以透過量測抗腫瘤細胞激素的含量，作為定義免疫系統狀態的指標。而測量中草藥對血液中細胞激素濃度的標準方法是使用酶聯免疫吸附試驗 (ELISA)。儘管ELISA具有靈敏性和特異性，但它程序較多，而且需要大量檢體才能做檢測。因此，在本研究中，利用微觀尺度下，粒子產生的布朗運動進行擴散度分析。根據斯托克斯-愛因斯坦方程，在恆定溫度和粘度下，粒徑與擴散度成反比，因此當粒徑增加時，粒子的布朗運動會變慢。在實驗過程中，將綠色螢光粒子表面修飾TNF-或IFN-捕獲抗體，並與檢體反應。再與表面有修飾TNF-或IFN-偵測抗體的聚苯乙烯粒子形成三明治免疫複合物。通過互相關算法分析粒子連續圖像以確定擴散率。在使用三明治免疫複合物測量校準曲線的實驗中，擴散係數隨著濃度的增加而降低，與預期相同。該系統建立的TNF-及IFN-小範圍校準曲線的檢測極限(LOD)為16.13 pg/mL。從中國醫藥大學獲得的臨床檢體僅需10 L即可測定，並花費5秒做量測、在2-3分鐘內完成分析。因此，本研究基於擴散法所開發的檢測系統可以實現低檢體量和高靈敏度的快速分析，用於中草藥治療之細胞激素濃度檢測。

Research shows that cancer is one of the top ten leading cause of death worldwide. In recent years, more and more people are suffering from cancer even at a much younger age. Cytokines are small proteins which play an important role in cancer pathogenesis. In tumor immunity, cytokines such as Tumor Necrosis Factor-alpha (TNF-a), Interleukin-2 (IL-2), and Interferon-gamma (IFN-) can inhibit tumor cell growth. While interleukin-10 (IL-10), interleukin-4 (IL-4) and other cytokines can promote the growth of tumor cells. According to research, Chinese herbal medicine can increase the production of anti-tumor cytokines, thereby improving the immune status in order to combat cancer formation and progression. In the experiment of detecting personalized Chinese herbal medicine recipe, the amount of anti-tumor cell cytokines can be measured as an index to define the state of the immune system. The conventional way of measuring the level of cytokines in the blood is by using enzyme-linked immunosorbent assay (ELISA). Although ELISA is sensitive and specific, it has many procedures and require large volumes of sample. Therefore, in this study, the Brownian motion generated by the particles at the microscopic scale is used to evaluate the diffusivity. According to the Stokes–Einstein equation, the particle diameter is inversely proportional to the diffusivity of the particles at a constant temperature and viscosity. Diffusivity of particles would be decreased when particle size increase. In the experiment, green fluorescent particles were functionalized with TNF- or IFN- capture antibodies and reacted were made to react with the serum sample. The samples were captured by polystyrene (PS) particles conjugated to TNF- or IFN- detection antibodies and forming the sandwiched immunocomplexes. The particle image sets were analyzed by the cross-correlation algorithm to determine diffusivity. In the experiment using the sandwich immunocomplex to measure the calibration curve, the diffusivity decreased with the increase of the concentration, which was the same as expected. The limit of detection (LOD) of TNF-and IFN- narrow range calibration curve established by this system is 16.13 pg/mL. The clinical specimen obtained from China Medical University (CMU) can be measured with only 10 L, and it takes 5 s to do the measurement, and the analysis is completed in 2-3 min. In conclusion, the detection system based on diffusometry that can achieve rapid analysis of low sample volume and high sensitivity, which is used for the detection of cytokine concentration in Chinese herbal medicine treatment.

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