醫學與健康科學

本研究利用 Verapamil 誘導斑馬魚胚胎心衰竭模式，並探討 Dapagliflozin 對斑馬魚胚胎表皮離子細胞的調控機制，以加深對 SGLT2 inhibitors 機制的了解。受精後第四天的斑馬魚在暴露於Verapamil 24小時後，除了抑制卵黃囊吸收以及造成心包膜水腫以外，對心臟整體功能(HR, EDV,ESV, SV, EF, CO)具負面影響。以粒線體染劑標記離子細胞，發現Verapamil使其密度上升，使用掃描式電子顯微鏡觀察，則可看到離子細胞頂端開口有明顯的萎縮，影響到正常功能。以抗體標記染色的方式檢測不同離子細胞亞型，顯示 Dapagliflozin 使富含 Na⁺-K⁺ ATPase 的 HR 細胞和富含 H⁺-ATPase 的 NaR 細胞密度上升。同時，心臟功能診斷標誌物的 mRNA 水平(naap, nppb,gata4, vmhc)暴露於Verapamil後上升，促進離子細胞代償性上調。

Cardiac Arrhythmia is one of the conditions in the group of heart and blood vessel diseases that can lead to sudden cardiac arrest (sudden death) and other conditions if not diagnosed quickly and accurately. According to research, heart and blood vessel diseases are the most common diseases and have a mortality rate of one-half of all non-communicable diseases. According to WHO statistics in 2012, it was found that there were 7.4 million deaths from heart and blood vessel diseases, and in 2017, the number of deaths increased to 177 million people, or about 94,444 people per day. Diagnosis of heart and blood vessel diseases can be done by measuring the electrical activity of the heart, and after the examination, a specialized physician will read and analyze the graph to find abnormal patterns. Currently, the shortage of qualified heart specialists to read the graph and screen for heart disease is a medical position shortage, which requires transferring data to hospitals with specialists, resulting in delays in diagnosis and treatment and even death. The project "EIPCA: Electrocardiogram Interpretation Pattern for Cardiovascular Abnormalities prediction" is an application program that assists in screening for fatal diseases that arise from abnormal heart rhythm. It employs artificial intelligence to aid in the screening and analysis of the electrical waveforms generated by an ECG machine, thus reducing diagnosis time and addressing the shortage of cardiology experts. EIPCA is comprised of two systems: (1) a system for screening and analyzing ECG waveforms using artificial intelligence to solve the problem of a shortage of specialized cardiology physicians, and (2) a system for risk assessment of fatal diseases by analyzing the ECG waveform data. The target group of the project is Rural hospitals, as well as health-related agencies. The project team hopes that the development of this project will significantly improve the efficiency and speed of screening for heart-related diseases, ultimately reducing the mortality rate from these diseases in the future.

上皮細胞黏附分子（EpCAM）與上皮細胞間黏附、信息傳導、增殖與分化等功能有密切關係，已被證實會在多種上皮癌細胞中大量表達，被視為一種可行的臨床標記。透過 細胞存活率、細胞群落、轉移與侵入試驗，觀察到EpCAM能增強未分化性甲狀腺癌（ATC）的細胞增殖、生長、轉移與侵入能力。 此外實驗發現dabrafenib小分子抗癌藥物處理的ATC，其細胞增殖、生長、轉移與侵入能力均有下降的趨勢，而細胞凋亡程度則有顯著的上升。此次研究藉由西方墨點法發現，磷酸化ERK蛋白的表現量隨dabrafenib濃度的上升而逐步下降，顯示dabrafenib能夠抑制ATC細胞訊息傳遞路徑中ERK蛋白的磷酸化，進而影響ATC的生長。若能深入了解EpCAM和dabrafenib在癌細胞中的作用機轉，EpCAM相關藥物與dabrafenib未來在臨床應用上，或許能為ATC患者提供另一種新的治療方式。

In recent years, lower inter-limb asymmetries have become a topic of increasing interest in sports research. Numerous studies have investigated the occurrence of between-limb differences in a variety of physical tests, ranging from strength, sprinting, and change of direction speed to jumping tasks. The main focus has been the association of asymmetries to either enhanced injury risk or reduced physical performance. Sport specific aspects and differences of lower limb asymmetries have not often been analysed. Additionally, most studies have been performed with male athletes. Women, however, present higher prevalence of lower limb asymmetries in strength, coordination, and postural control than men. These two aspects were therefore addressed in the present study. Female youth soccer (n=18, age: 16.7 ± 0.8 years) and floorball (n=18, age: 17.6 ± 0.9 years) players completed a test battery consisting of six unilateral jumping tasks in horizontal and lateral direction to detect sport specificity in inter-limb asymmetries. The test comprised the following hops: (1) Single Leg Hop for Distance and (2) a newly created version of it, (3) Single Leg Triple Hop, (4) Single Leg Crossover Hop, (5) Side Hop and (6) Single Leg 6-meters Timed Hop. The scores of every jump were calculated into Limb Symmetry Indexes for each participant. A linear mixed effect (LME) model (using function lmer in program R) was applied to evaluate the effects of sport and jump type on asymmetries. In terms of the whole test battery, there was a significantly higher magnitude of asymmetries in soccer compared to floorball (p=0.0067) with a mean difference of 1.9%. Three significant differences between the effects of different jump types were detected (5>2: p=0.027; 5>3: p6: p=0.014). Moreover, the results showed no significant effect on leg dominance. According to the findings of this study, soccer appears to be more asymmetric than floorball, leading to the suggestion that inter-limb asymmetries may not only be task-specific, but also sportspecific. Since no significant correlations were detected, this study suggests that inter-limb asymmetries are independent of leg dominance. Considering the possible reduction in athletic performance and increasing injury risk, strength and conditioning coaches are advised to assess athletes' inter-limb asymmetries using a broad, sport specific test battery and decrease them.

肝細胞癌 (HCC) 為全球導致高死亡率的癌症之一，第一線標靶治療藥物 Sorafenib 雖能延長患者存活期，但其療效有限且伴隨嚴重副作用。在癌症中，中心體異常所導致的染色體變異是腫瘤發展的關鍵因素，而動力蛋白已知參與中心體裝配，且前人研究結果表明動力蛋白與肺癌、 HCC 等多種癌症有關連 。故本研究先透過基因表現資料庫分析，發現 HCC患者中的動力蛋白重鍊基因表現量大致顯著高於一般，後以 Ciliobrevin D 抑制三種 HCC 細胞株 Hep3B、HepG2、Huh-7 中的動力蛋白，並藉細胞存活率分析、遷移試驗與西方墨點法，探討抑制動力蛋白與 HCC 的關聯。據實驗結果，抑制動力蛋白後， Huh-7 的遷移速率減緩， 蛋白質表現量亦隨抑制劑濃度升高而降低。這表示抑制動力蛋白具有抑制肝癌細胞轉移的潛力，期未來能成為肝癌新的治療靶點。

先前研究發現一種多元不飽和脂肪酸 15-keto-PGE2 能減少肌肉慢性發炎以及增加肌肉生長因子胰島素的敏感性，具有增加肌肉的潛力。我們探討此脂肪酸在小鼠中將脂肪組織轉成肌肉組織的可能性。 我們先以 15-keto-PGE2 治療肥胖型肌少症小鼠後，再以全基因組mRNA定序，發現在小鼠皮下脂肪中，肌肉特異基因表現量高度上升。透過基因表現路徑分析軟體 GSEA證實此脂肪酸會高度引發皮下脂肪中，與肌肉分化、肌肉收縮與肌肉結構相關的生物路徑。另外， 15-keto-PGE2 也可以誘導脂肪前驅細胞3T3-L1中與肌肉生成相關的基因表現。 最後，我們發現以 15-keto-PGE2 治療的老年肥胖型肌少症小鼠，體重、血糖、脂肪比例下降，肌肉質量及力量上升，證實了它在體內同時減少脂肪並增加肌肉量的效果。

亨丁頓舞蹈症為與認知功能障礙密切相關的神經退行性疾病。本研究首次應用動態葡萄糖強化磁振造影（DGE MRI）以了解葡萄糖代謝作為亨丁頓舞蹈症神經影像生物標記的可行性，以分析大腦中不同區域之間的代謝關係。 本研究對腦區間葡萄糖代謝關聯性進行分析，並針對訊號進行自動化分群，觀察特定訊號樣態之特徵。於zQ175 KI和R6/2 KI小鼠中不同的連接性變化模式中，發現紋狀體和齒狀回之間葡萄糖代謝連接性具顯著變化，與已知病理一致，顯示DGE MRI作為臨床生物標記之潛力，以利及時診斷和監測該疾病。 這項開創性的研究探索了使用DGE MRI作為亨丁頓舞蹈症影像標記可行性，並詳細分析腦區間葡萄糖代謝相關性，不僅進一步對該疾病之病理更加深入了解，同時提高早期診斷、疾病監測和精準醫療應用發展，說明可能有針對代謝紊亂的潛在治療策略。

遺傳性表皮分解性水泡症（EB）是種罕見疾病，因突變使角蛋白異常，造成表皮組織脆弱易形成水泡，單純型水泡症（EBS）是最常見的類型。此計畫旨在：（一）建立果蠅EBS疾病模型；（二）探討溫度對病徵的影響；（三）以此果蠅EBS疾病模型發展藥物篩選平台。初步使用Diacerein測試，評估對EBS症狀的改善效果。 目前顯示突變角蛋白K5/K14R125C會形成積聚體，與正常K5/K14形成的角蛋白網絡不同；全程25℃培養，約32%果蠅翅膀有水泡，亦符合EBS病徵。篩藥平台建置已完成色素和溶劑DMSO劑量測試，初步顯示Diacerein有助病徵緩解。目前將擴大統計不同溫度對 EBS果蠅死亡率、水泡發生率和角蛋白積聚形成比例。希望以本研究建立的果蠅EBS疾病模型與篩藥平台，能為罕見遺傳疾病療程開發奠定基礎。

Calcineurin (CN) activation is a main cause of cancerous tumor formation, one of the leading causes of death globally. Cyclosporine-A (CsA) is a commercially available oral drug that inhibits CN activation; however, low bioavailability limits its use. Nine patented CsA conjugates are potential alternatives to CsA as they have improved cytotoxicities and bioavailabilities but unknown CN-binding affinity. This study aimed to identify the CNinhibition strength and bioavailability of CsA conjugates in silico drug-likeness evaluation via modified Lipinski’s Rule of Five was done on CsA, voclosporin, and CsA conjugates to test bioavailability. The binding affinities of bioavailable compounds were computed via docking to CN in five trials, and the binding affinities were compared. The Water-soluble, RVal, IIA, Alpha, and MeBmt 2 conjugates showed improved bioavailabilities compared to CsA as they passed the drug-likeness screening. After five trials of computational docking to CN, the IIA and RVal conjugates showed improved binding affinities at -15.8 kcal/mol and -15.2 kcal/mol, respectively, compared to CsA at -14.3 kcal/mol. Notably, IIA also showed an improved binding affinity compared to voclosporin at -15.5 kcal/mol. These results suggest that CsA conjugates may be better oral chemotherapeutic drugs than CsA.

本研究以人類肺癌A549細胞株和纖維母細胞模擬體內腫瘤微環境，挖掘纖維母細胞如何促進癌細胞的生長。從病人的正常和癌組織提取癌相關纖維母細胞（cancer-associated fibroblasts, CAFs） 和 正常纖維母細胞（Normal Fibroblasts, NFs），經過基因序列一對對作分析，開發新的治療策略和潛在的靶點。利用核糖核酸定序（RNA-Seq）分析發現CAFs會比NFs分泌更多SNARE 蛋白 YKT6，而更深入地探究獲悉YKT6會透過活化YKT6+CAFs途徑促進肺癌A549細胞惡化，此惡化過程包括誘導及提升癌細胞的生殖（proliferation），轉移(migration)和入侵(invasion)能力。 此外，在 CAFs 中敲除 YKT6基因，減弱CAFs 的外泌體（exosome）釋放，從而調節了其對肺癌細胞A549的腫瘤促進作用。本研究發現靶向YKT6並抑制外泌體分泌，從而降低CAFs對肺腺癌細胞的腫瘤支援功能可以為肺癌治療提供一種新的策略。