醫學與健康科學

溫熱化學治療(Hyperthermic Intraperitoneal Chemotherapy, 簡稱HIPEC)，為將化療藥物加熱後灌注到腹腔內殺死癌細胞，但溫熱化學治療只能在開刀時使用，治療次數有限。為達到多次的熱化療效果，本研究開發奈米氧化鐵藥物磁性複合微粒(Nano iron oxide magnetic drug complex particles, NIOMP)，可利用開刀時包覆複合微粒於腹腔，術後用磁場加熱，加上微粒化療藥物釋放，達到多次溫熱化學治療。NIOMP以鐵氧化物奈米粒子為核心，海藻酸鈉為外殼體，利用電紡法將前其與氯化鈣交聯產生凝聚性微粒。數據顯示最佳製作條件為16 kv高電壓、距離4 cm、噴速5μm/s，可得直徑大小約為200~450 μm穩定性高微粒。藥物釋放實驗顯示，NIOMP於2~7小時藥物釋放速率較穩定。利用可變磁場之電磁效應加熱NIOMP中氧化鐵磁顆粒，可於30分鐘內快速升溫至43℃。細胞相容性測試結果得知，NIOMP對細胞無毒性。本研究改善HIPEC的治療限制，可多次針對腫瘤患部局部熱化學治療，未來將可改善病人存活率。

本研究探討高壓氧治療對動物傷口的進程時間與皮膚創口的癒合品質。使用自行開發之動物高壓氧艙(1.4~3ATA)，幫助流浪動物結紮傷口快速癒合，並救助傷口嚴重潰爛，瀕死的流浪動物。本研究選30隻體態相近之流浪貓，進行節育手術後隨機抽取15隻貓作為試驗組，其餘15隻貓作為對照組。試驗組在常規治療的基礎上，每天再採用1個小時的高壓氧治療，對照組僅採用常規治療，持續觀察兩組試驗動物的傷口癒合進程，其結果顯示試驗組無論是傷口癒合速度或瘢痕組織的平整度皆優於對照組，瀕死動物救治實驗，成功救活無法手術的流浪動物。

缺氧（Hypoxia）在癌症中，影響腫瘤微環境（Tumor microenvironment，TME）的重塑扮演重要的角色。原位癌細胞與欲轉移之遠端器官的微環境間，會透過細胞外囊泡（extracellular vesicles，EVs）進行細胞間相互作用，對於轉移前腫瘤微環境（Pre-metastatic niche，PMN）的形成和轉移過程至關重要。 本研究目的在闡明缺氧條件下，肺腺癌如何透過產生的外囊泡重塑肝臟轉移前微環境。結果顯示缺氧刺激肺癌產生的細胞外泌體（Exosome）增加了肝臟星狀細胞（hepatic stellate cell，HSC）活化和血管生成。由於缺氧肺癌所分泌的EVs含有CD151蛋白，促進HSC的活化和血管新生。缺氧肺癌所產生的Exosome增 加了HSC和內皮細胞中FAK（focal adhesion kinase）、Src（SRC proto-oncogene, non-receptor tyrosine kinase）和AKT（AKT serine/threonine kinase）通路的激活。 總結此實驗證實了缺氧腫瘤衍生的Exosomes，探討其中富含的CD151機制，活化了HSC與特別的訊息傳遞於細胞中，調節肝臟PMN的形成。因此CD151可作為肺腺癌轉移至肝臟的新治療靶點。

登革熱是一種由蚊子傳播的全球性疾病，已在全球各國造成了嚴重破壞。當前沒有針對登革熱的疫苗或療法，使得早期診斷階對該疾病的預防極為重要。本實驗提出了一種新的診斷模型：自旋增強型側向免疫測定。自旋增強側流免疫分析平台與診斷標記物螢光奈米鑽石結合使用，利用電磁場調控螢光奈米鑽石發出的信號，以提供準確和靈敏的結果，取代傳統的奈米金側向免疫測定平台。我們發現基於螢光奈米鑽石的自旋增強型側面免疫分析法在登革熱病毒診斷中的應用不僅可以準確地檢測出登革熱病毒抗原NS1，而且還提供了比基於奈米金的橫向流免疫分析法高約100倍的信號靈敏度。自旋增強型側向免疫測定法是針對登革熱病毒的一種改進的診斷工具， 未來可以應用於其他病毒篩檢，例如寨卡病毒和新型冠狀病毒。

Commercially available bandages such as hydrocolloid are neither biodegradable nor anti-bacterial. Chitin is known to be the second most naturally available polysaccharide which could be transformed to chitosan which is known to be anti-bacterial (Hasan, 2018) (Chao, 2019) and haemostatic (Okamoto, 2003) (Hu, 2018). Chitosan can be further converted to hydrogel which is bio-degradable and has good water absorbance. Anti-bacterial crab bio-bandages and crab bio-dressings should be bio-degradable as it took 42 days and a month for complete bio-degradation respectively, so they should be better than commercial bandages such as Nexcare Hydrocolloid as the disposal of anti-bacterial crab bio-bandages with bio-dressings would no longer pose burden to landfilling or threat to our environment. Anti-bacterial crab bio-bandages with bio-dressings are anti-bacterial with degree of deacetylation of DD% (measured using FTIR Spectrum II) 82.6% (due to the presence of chitosan) even without the application of other anti-bacterial agents and hence can provide complete protection of wounds from skin and soft tissues infections and haemostatic (due to the presence of chitosan). After testing and certification based on IS997:2004 and BS EN 13726-1, they should meet many requirements specified. Anti-bacterial crab bio-bandages should be eligible for marketing. Some results were as follows: 1.4 Anti-bacterial effect of crab hydrogels and roasted crab hydrogels Pure chitosan, crab chitosan, crab hydrogels and roasted crab hydrogels showed significant anti-bacterial effect. NO oral bacterial colonies were present in drinking water with crab hydrogels. Thus crab hydrogels could serve as effective anti-bacterial wound dressings. 1.6 Basing on IS997:2004 standard, the load per unit of area of anti-bacterial bio-bandages was 342g/m2 which met the minimum requirement of 36g/m2, the anti-bacterial bio-bandages had stronger tension strength (>20N both in dry and wet conditions) than commercial hydrocolloid. (2.7N dry 2.8N wet) which was comparable with that required (50-67N) and pH of about 7 which met the pH range of 4.5-8. 1.7 The FSA Free-Swell Absorbency of synthetic blood of crab hydrogel bio-dressings was 1.86g per 5cm x 5cm dressing which was much higher than that of commercial hydrocolloid (0.299g per 5cm x 5cm dressing) based on BS EN 13726-1.

Swimming pools are one of the transmission routes of superficial and cutaneous fungal infections. Maintenance of environmental hygiene in different parts of swimming pools is of great importance, especially the hygiene of water (1). The conventional fungal detection methods include direct smear preparation, culture, and pathological examinations. However, these methods are not fast enough or do not have sufficient sensitivity (2). Therefore, the present research introduces a novel method for detecting Aspergillus niger in pool water through creating optimal conditions for this fungus, which leads to the citric acid production by the fungus and pH changes of the related culture medium. Four experiments in 10 steps were performed to find the optimal conditions for fungal growth. According to our results, adding each of the variables sucrose, soy, and ferrous sulfate can lead to favorable results. Moreover, the shaker speed increase and fungal aeration are important. Also, we showed that soybean led to the best results compared to other variables. Considering the obtained results, including the shortened detection duration and cost-effectiveness, this method can be presented to the swimming pool owners and pathobiology laboratories as the method of choice for Aspergillus niger detection.

肺癌為全球最常被診斷的癌症之一，主要分為非小細胞肺癌與小細胞肺癌，其中小細胞肺癌是最具侵略性、致死性且預後最差的肺癌。目前針對小細胞肺癌仍未有有效的療法。 本研究探討花生外皮萃取出之山奈酚醣苷衍生物 kaempferol 3-O-β-d-xylopyranosyl-(1→2)-α-l-rhamnopyranosyl-7-O-α-l-rhamnopyranoside 對小細胞肺癌的抗癌功效。我們發現細胞株H1688經藥物處理後細胞存活率明顯降低，以流式細胞儀細胞週期分析，隨藥物劑量增加時 sub G1 期細胞數明顯上升；以 Annexin V 觀察發現藥物劑量增加時細胞逐漸走向晚期凋亡，此外，隨劑量增加 c-PARP、caspase-3、-3和 -9 的蛋白表達都有增進，同時也造成粒線體膜電位下降與 cytochrome c 釋放，另外我們透過西方墨點法分析發現隨著藥物劑量的增加，p53蛋白的表現量下降且有磷酸化的現象，並且在經過咖啡因的處理後藥物對細胞株的毒殺作用更加明顯，由實驗結果可知kaempferol 3-O-β-d-xylopyranosyl-(1→2)-α-l-rhamnopyranosyl-7-O-α-l-rhamnopyranoside能夠良好的誘導H1688細胞凋亡，有潛力成為治療小細胞肺癌的藥物。

Swimming pools are one of the transmission routes of superficial and cutaneous fungal infections. Maintenance of environmental hygiene in different parts of swimming pools is of great importance, especially the hygiene of water (1). The conventional fungal detection methods include direct smear preparation, culture, and pathological examinations. However, these methods are not fast enough or do not have sufficient sensitivity (2). Therefore, the present research introduces a novel method for detecting Aspergillus niger in pool water through creating optimal conditions for this fungus, which leads to the citric acid production by the fungus and pH changes of the related culture medium. Four experiments in 10 steps were performed to find the optimal conditions for fungal growth. According to our results, adding each of the variables sucrose, soy, and ferrous sulfate can lead to favorable results. Moreover, the shaker speed increase and fungal aeration are important. Also, we showed that soybean led to the best results compared to other variables. Considering the obtained results, including the shortened detection duration and cost-effectiveness, this method can be presented to the swimming pool owners and pathobiology laboratories as the method of choice for Aspergillus niger detection.

上皮細胞黏附因子 (EpCAM)參與了細胞的黏附、信息傳遞、增殖及分化等功能，並在惡性腫瘤組織中大量表達，被當作各種癌症的診斷標誌物，因此被認為是治療癌症的重要關鍵。另外近期研發的抗EpCAM中和性抗體被認為能抑制EpCAM的訊息傳遞，導致腫瘤細胞的死亡，也可以降低癌細胞中PD-L1蛋白的表現，進而導致腫瘤細胞死亡及活化T細胞殺死癌細胞的能力。 為了觀察EpCAM是否會對癌細胞造成影響，我們將癌細胞分成野生型 (wild type)以及EpCAM基因剔除細胞株，並以Western blotting及qRT-PCR確認實驗組的EpCAM基因有確實被成功剔除。進一步利用細胞存活率及細胞群落實驗證實EpCAM會促進大腸直腸癌細胞增生，並以抑制劑DAPT和TAPI-1處理證實EpCAM訊息傳遞會增加癌細胞轉移與侵入能力。接著為了驗證EpCAM中和性抗體的作用，我們用EpCAM中和性抗體處理癌細胞後，由細胞凋亡實驗確認EpCAM中和性抗體確實會促使癌細胞凋亡。我們的研究結果顯示EpCAM的訊息傳遞會促進腫瘤增生與轉移，而EpCAM中和性抗體於癌症治療中的高度前瞻性，期許將來能在癌症治療中提供一個可行有效的療法。

本研究以腸道類器官(organoid)模擬體內環境，分析乳癌標靶藥物Lapatinib與Tucatinib對腸道產生副作用的差異。Lapatinib明顯抑制ileum及colon organoid的形成，其IC50低於Tucatinib約1000倍。其中Lapatinib特別對adult type organoid較具明顯抑制作用，顯示影響腸道上皮細胞的分化功能。以RNA seq 與Ingenuity pathway Analysis分析藥物對organoid中轉錄體表現的影響，Lapatinib 在colon organoid中增加腸道發炎、葡萄糖代謝異常、氯離子外流等基因群的表現，並降低crypt發展的基因群。其中，Lapatinib藉由增加Glut3的表現提高organoid對葡萄糖的吸收，此作用受到L-ascorbic acid (Vitamin C)抑制，亦增加GABA receptor 提高氯離子外流，顯示代謝與電解質失衡及發炎作用可能為lapatinib造成腹瀉的主因之一。以3D organoid為可信賴的轉譯研究模式，我們發現同屬HER2 tyrosine kinase inhibitor的Lapatinib與Tucatinib對腸道功能產生迥然不同的影響，並發現合併使用Glut3 inhibitor或GABA receptor antagonist可能可成為減緩Lapatinib副作用的對應策略。