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An Integrated Approach For Uncovering Novel DNA Methylation Biomarkers For Non-small Cell Lung Carcinoma

為了解決Ac 504van tw的問題，作者VAIBHAV KUMAR SUNKARIA 這樣論述：

Introduction - Lung cancer is one of primal and ubiquitous cause of cancer related fatalities in the world. Leading cause of these fatalities is non-small cell lung cancer (NSCLC) with a proportion of 85%. The major subtypes of NSCLC are Lung Adenocarcinoma (LUAD) and Lung Small Cell Carcinoma (LUS

C). Early-stage surgical detection and removal of tumor offers a favorable prognosis and better survival rates. However, a major portion of 75% subjects have stage III/IV at the time of diagnosis and despite advanced major developments in oncology survival rates remain poor. Carcinogens produce wide

spread DNA methylation changes within cells. These changes are characterized by globally hyper or hypo methylated regions around CpG islands, many of these changes occur early in tumorigenesis and are highly prevalent across a tumor type.Structure - This research work took advantage of publicly avai

lable methylation profiling resources and relevant comorbidities for lung cancer patients extracted from meta-analysis of scientific review and journal available at PubMed and CNKI search which were combined systematically to explore effective DNA methylation markers for NSCLC. We also tried to iden

tify common CpG loci between Caucasian, Black and Asian racial groups for identifying ubiquitous candidate genes thoroughly. Statistical analysis and GO ontology were also conducted to explore associated novel biomarkers. These novel findings could facilitate design of accurate diagnostic panel for

practical clinical relevance.Methodology - DNA methylation profiles were extracted from TCGA for 418 LUAD and 370 LUSC tissue samples from patients compared with 32 and 42 non-malignant ones respectively. Standard pipeline was conducted to discover significant differentially methylated sites as prim

ary biomarkers. Secondary biomarkers were extracted by incorporating genes associated with comorbidities from meta-analysis of research articles. Concordant candidates were utilized for NSCLC relevant biomarker candidates. Gene ontology annotations were used to calculate gene-pair distance matrix fo

r all candidate biomarkers. Clustering algorithms were utilized to categorize candidate genes into different functional groups using the gene distance matrix. There were 35 CpG loci identified by comparing TCGA training cohort with GEO testing cohort from these functional groups, and 4 gene-based pa

nel was devised after finding highly discriminatory diagnostic panel through combinatorial validation of each functional cluster.Results – To evaluate the gene panel for NSCLC, the methylation levels of SCT(Secritin), FOXD3(Forkhead Box D3), TRIM58(Tripartite Motif Containing 58) and TAC1(Tachikinin

1) were tested. Individually each gene showed significant methylation difference between LUAD and LUSC training cohort. Combined 4-gene panel AUC, sensitivity/specificity were evaluated with 0.9596, 90.43%/100% in LUAD; 0.949, 86.95%/98.21% in LUSC TCGA training cohort; 0.94, 85.92%/97.37 in GEO 66

836; 0.91,89.17%/100% in GEO 83842 smokers; 0.948, 91.67%/100% in GEO83842 non-smokers independent testing cohort. Our study validates SCT, FOXD3, TRIM58 and TAC1 based gene panel has great potential in early recognition of NSCLC undetermined lung nodules. The findings can yield universally accurate

and robust markers facilitating early diagnosis and rapid severity examination.

開發米與幾丁質減積製程並提升酵素降解速率

為了解決Ac 504van tw的問題，作者陳衍齊 這樣論述：

米與幾丁質經酵素降解可得葡萄糖及N-Acetyglucosamine(GlcNAc)，在醫療技術上及營養層面皆展現非比尋常的價值，而粒子微小化可幫助其降解速率增加。本研究將米與幾丁質兩種生質原料經由兩階段磨碎，得到所需粒徑尺寸，並驗證其酵素反應的提升。於不同的機台進行物料尺寸的微小化時，物料的物化特性或是機台本身的參數設定都會影響機台將物料尺寸微小化的效率。於第一階段乾式切削時，由實驗設計及反應曲面法求得米在含水率 1.2 %、切削轉速17918 rpm及切削時間3 min時為最佳化操作參數；幾丁質在含水率5.5 %、切削轉速17837 rpm及切削時間6.4 min時為最佳操作參數。於第二

階段濕式研磨時，第一段以研磨轉速1400 rpm、研磨間距50 µm 及研磨時間1.5 hr，第二段以研磨轉速1400 rpm、研磨間距30 µm 及研磨時間4 hr 為最佳操參數，其平均粒徑達5.1 µm ；幾丁質於研磨轉速1400 rpm、研磨間距5 µm及研磨時間12 hr時為最佳操作參數，其平均粒徑達22.1 µm。另外於酵素反應下檢測反應速率變化，由Michaelis-Menten動力學方程式得知，在最佳操作參數下觀察米的粉體研磨情形，V_max提升11.5倍，於長時間反應下轉化率提升36倍；在最佳操作參數下觀察幾丁質粉體研磨情形，V_max提升26.1倍，於長時間反應下轉化率提升3

2.2倍。