全國中小學科展

醫學與健康科學

由誘導性多功能幹細胞篩選新生抗原用於大腸直腸癌之抗癌應用

癌症治癒方式成效有限,在iPSC中找出具有潛力的蛋白製成大腸直腸癌疫苗是本研究目的。 實驗先以C57BL/6小鼠之股骨及脛骨骨髓中造血幹細胞分化成為未活化的樹突細胞。放入安慰劑、CpG佐劑、CpG+MC38、CpG+iPSC等抗原物來進行免疫細胞活化測試,流式細胞儀分析結果驗證能否減緩小鼠大腸直腸癌的腫瘤生成。接著,利用質譜儀及大數據分析從iPSC中找出促使樹突細胞活化的抗原物蛋白成分,選出3個候選,透過西方墨點法確認其專一性。查詢候選蛋白TTW1、2、3在The Human Protein Atlas 網站對大腸直腸癌之表現數據。 結果 iPSC的抗原物促使樹突細胞的MHC1標記物活化,助於毒殺性 CD8+T細胞的產生進而透過免疫系統預防大腸直腸癌的產生。TTW1只有在iPSC細胞株表現出專一性,同時資料庫顯示TTW1在大腸直腸癌中具有高表現量。 目前未有文獻探討抗原疫苗的應用,團隊未來專注合成iPSC中TTW1蛋白,進一步開發成大腸直腸癌疫苗。

原薯蕷皂苷對腎臟癌細胞的影響

癌症治療多半會傷害到人體的健康,所以國民大多較偏好以較養生的方法來治療癌症,例如中藥。本實驗以山藥萃取物原薯蕷皂苷抑制腎癌細胞 A498 及 786-O ,期望能達抑制腎臟癌細胞增生之目的。 實驗方法包括以 MTT 試驗、細胞菌落試驗來觀測腎癌細胞受原薯蕷皂苷作用後的活性及存活量,再透過西方墨點法及流式細胞儀來了解腎臟癌細胞死亡途徑。實驗結果顯示將原薯蕷皂苷抑制人類腎癌細胞株786-O及A498的增生能力具有抑制的能力。再透過流式細胞儀的分析,顯示原屬蕷皂苷可誘發兩種腎臟癌細胞的凋亡作用,並且透過西方墨點法觀察出是抑制Bcl-2蛋白、增加Bax蛋白和caspase-9/PARP蛋白的表現,進而導致腎臟癌細胞株產生細胞凋亡。 本研究是在實驗室中進行,且只是利用細胞株來觀測此項研究結果。或許未來可以透動物實驗以及臨床實驗,確認原薯蕷皂苷抗癌之功效,並推廣至全球以造福全人類之健康。

UVB induced TRPV1 and TRPA1 expression in skin keratinocyte and dorsal root ganglion cells: a plausible cause of warm and pain by sunlight irradiation

陽光晒到皮膚,人體會感覺到刺痛以避免過強紫外線的曝露。但皮膚為何會有刺痛的感覺呢?過去對光的生體受器著重於眼睛錐及桿狀細胞(Opsin 1/2),但它們在背根神經元 (DRG,周邊神經末端主體細胞)的表現並不多。TRP channels表現在皮膚及神經。TRPV1是一種痛覺受器,活化時會有鈣離子通透,Dr. Julius因它得到2021諾貝爾獎。TRPV1/A1可受溫度、酸度等活化,但DRG細胞表現的TRPV1/A1,是否會因紫外線(UVB)照射而影響,並導致鈣通透,目前並不清楚。我以不同強度UVB照射人類角質細胞或大鼠DRG,以螢光顯微鏡及流氏細胞儀測量TRPV1/A1蛋白質表現,以實時影像來作動態鈣離子分析。結果顯示UVB在10 mJ/cm2可增加DRG的TPRA1/V1表現,但UVB在5mJ/cm2照射DRG細胞後,只增加TRPA1而不是TPRV1的表現。在皮膚的角質細胞,不管是使用螢光顯微鏡或是流氏細胞儀,UVB在20mj/cm2以上的能量強度會造成角質細胞的毒性,以10mJ/cm2 UVB照射角質細胞,則會增加角質細胞的TPRA1及TRPV1表現,其中又以TRPA1的增加較明顯。惟UVB照射對細胞鈣離子通透的實時影響不大。我的結論是,UVB照射增加角質細胞及DRG的TRPA1/V1表現(特別是TRPA1),這些改變可能與光照引起之麻痛有關。

基因突變與骨硬化症之相關

本研究由骨硬化症(osteosclerosis)個案報告開始,進行基因變異的探查,先針對骨生成主要調控機制WNT pathway上的SOST、LRP5、LRP6基因,進行基因定序與分析。在此3個基因沒有發現異常位點後,轉而以全外顯子組定序(whole exome sequencing),進行次世代基因定序(next generation sequencing)。結果顯示病患帶有CTNNB1 c.1982G>A (p.Arg661Gln)之異合子(heterozygous)誤義變異(missense mutation),並證實是CTNNB1基因的原發突變(de novo mutation),該突變影響後續蛋白質表現,影響了ICAT對WNT/beta-catenin訊息傳遞鏈的抑制,繼而穩定beta-catenin,造成骨質異常增生、並導致全身骨硬化。此案例為全世界第二例、亞洲首例的CTNNB1基因功能增強突變(gain-of-function mutation)的病例報告。

上皮細胞黏附因子(EpCAM)對腫瘤增生影響之機制探討

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。另外抗EpCAM中和性抗體可以阻斷EpCAM訊息傳遞,進而引起癌細胞PD-L1的表現量降低並促進T細胞的毒殺活性。為了觀察EpCAM是否會對癌細胞的增生、轉移以及侵入能力造成影響,我們將細胞分為野生型 (wild type) 和EpCAM基因剔除細胞株 (EpCAM knockout) 進行實驗。首先,我們以Western-blotting和qRT-PCR確認EpCAM基因剔除組的EpCAM基因有確實被剔除,再進一步利用細胞存活率及細胞群落實驗證實EpCAM會促進癌細胞的增生能力,並分別藉由EGFR的抑制劑Afatinib與HGFR的抑制劑Crizotinib對於癌細胞存活率的實驗證實EpCAM 對於EGFR與HGFR的訊息傳遞扮演重要角色。接著分析EpCAM對癌細胞轉移及侵入能力的影響,γ-secretase和ADAM17皆為裁剪EpCAM進行訊息傳遞的重要酵素,實驗中的癌細胞分別以γ-secretase和ADAM17的抑制劑DAPT和TAPI-1處理,證實EpCAM的訊息傳遞與癌細胞轉移與侵入能力有關。接著我們以EpCAM中和性抗體處理癌細胞後,證實EpCAM中和性抗體確實會促使癌細胞凋亡。最後我們利用Western blotting分析EpCAM對於不同激酶磷酸化的蛋白質表現量之影響,找出EpCAM下游訊息傳遞的完整路徑,期望能找到治療癌症的關鍵。

Silver Moringa Cloth: Silver Nanoparticle Fabric Based on Moringa Extract (Moringa oleifera) as Antibacterial Against Methicilin Resistant Staphylococcus aureus

Staphylococcus aureus is addressed as one of the most common pathogens in hospital settings and in the community. This pathogen causes invasive infections, sepsis, and death. The emergence of antibiotic-resistant bacteria is due to bacterial mutations and the use of antibiotic drugs that are not by procedures. Resistance makes MRSA infections difficult to treat, resulting in high healthcare costs. These problems lead to an urgent need to find alternative drugs to control MRSA infection. Therefore, developing new drugs and procedures such as antibacterial nanoparticles, are particularly promising. Indonesia has many medicinal plants with antibiotic activity, including Moringa oleifera. Moringa oleifera contains several active compounds such as alkaloids, flavonoids, and saponins which are known to have antibiotic activity. Silver nanoparticles or AgNPs are currently used as antimicrobial agents because they are toxic to prokaryotic cells (bacteria) but relatively safe for eukaryotic cells. AgNP synthesis mediated by M. oleifera extract has the advantages of being non-toxic, pollution-free, and environmentally friendly. Sisal is a potential source of naturally derived fabric and a prospective source of multifunctional textiles. Recent studies have utilized and functionalized sisal to develop composite materials. However, functionalizing of sisal using nanosilver-based materials has not been studied yet. Bioactive chemicals from plant-extracted nanoparticles also provide additional antimicrobial properties. This study aims to produce AgNPs mediated by M. oleifera leaf extract and to analyze its antimicrobial effect on MRSA growth. The powdered Moringa (4g) was boiled with 100 ml of distilled water (550 C) for 15 minutes. The mixture was filtered through Whatman No 1 filter paper and store refrigerated. The nanoparticle was synthesized by rinsing sisal fabric cloth to several concentrations of AgNO3 (1mM, 10mM, and 20mM) with Moringa extract. Nanoparticle synthesis from AgNO3 done with the help of Moringa oleifera extract. The resulting AgNPs have MIC values (Minimum Inhibitory Concentration) and MBC (Minimum Bacteriocidal Concentration) of 1.25 mg/ml. The resulting silver nanoparticles showed antibiotic activity against MRSA with an average inhibition zone diameter of 15.677 mm. XRD and SEM studies are going to be held to support the data.

Analysis of lncRNA, miRNA , mRNA-associated ceRNA networks include in promoting glioma cancer

The ceRNAs as a class of RNAs act by competitively binding to miRNAs and limiting their regulatory effect on the target genes. Increasing evidences point to the role of ceRNAs in glioma cancer. So far, limited studies have been reported on the role of ceRNA in the development in glioma cancer. In this study, we have analyzed online RNA sequencing data in order to predict the ceRNAs which are putative regulators of in glioma cancer.

Overcoming Motion Disorders

Helping in overcoming the motion disorders is an important thing which deserve striving for. Every year number of people who have MS or paresis (paraparesis) is increasing. The device has effective results for the patients and it backs to finding the conditions of natural treatments in the device. Obviously, this device helps them to keep standing and achieve the required exercises the movement like normal people by using their hands. The degree of improvement differs from person to another as depends on the hardness of the disease and injury kind.