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Of All Things

為了解決v11 absolute的問題，作者Benchley, Robert C. 這樣論述：

OF ALL THINGS. Originally published in 1921. PREFACE: WHEN, in the Course of human events, it becomes necessary for one people to dissolve the political bands which have connected them with another, and to assume among the powers of the earth, the separate and equal station to which the Laws of Natu

re and of Natures God entitle them, a decent respect to the opinions of mankind requires that they should declare the causes which impel them to the separation. We hold these truths to be self-evident, -that all men are created equal, that they are endowed by their Creator with certain unalienable R

ights that among these are Life, Liberty and the pursuit of Happiness. That to secure these rights, Governments are instituted among Men, deriving their just powers from the consent of the governed, -That whenever any Form of Government be- comes destructive of these ends, it is the Right of the Peo

ple to alter or to abolish it, and to institute new Government, laying its foundation on such principles and organizing its powers in such form, as to them shall seem most likely to effect their Safety and Happiness. Prudence, indeed, will dictate that Governments long established should not be chan

ged for light and transient causes and accordingly all experience hath shewn, that mankind are more disposed to suffer, while evils are suf- ferable, than to right themselves by abolishing the forms to which they are accustomed. But when a long train of abuses and usurpations, pursuing invariably th

e same Object, evinces a design to reduce them under absolute Despotism, it is their right, it is their duty, to throw off such Government, and to provide new Guards for their own future security. Such has been the patient suf- ferance of these Colonies and such is now the necessity which constrains

them to alter their former Systems of Government. The history of the present King of Great Britain is a history of repeated injuries and usurpations, all having in direct object the establishment of an abso- lute Tyranny over these States. To prove this, let Facts be submitted to a candid world. R.

C. B. The Rookery Breerning Downs Wippet-cum-Twyne New York City August 24 1921. COntents include: CHAPTER PAGP I THE SOCIAL LIFE OF THE NEWT . . 3 I1 COFFEE, MEGG AND ILK, PLEASE 10 I11 WHEN GENIUS REMAINED YOUB HUMBLE SERVANT . . . . . 18 IV THE TORTURES OF WEEK-END VISIT- I N G . . . . . ., . .

. 32 V GARDENING NOTES . . ., . V1 LESSONNUMBERONE .. L 43 . . . . 5. V11 THOUGHTS ON FUEL SAVING . . . 65 V111 NOT ACCORDING TO HOE . . . 77 IX FROM NINE TO FIVE . ., . . . 89 X TURNING OVER A NEW LEDGER LEAP 102 XI A PIECE OF ROAST BEEF . . . . 110 XI1 THE COMMUNITY MASQUE AS A SUB- STITUTE FOR WA

R . . . . 121 XI11 CALL FOR MR. KENWORTHY . . X30 XIV FOOTBALL COURTESY OF MR. MORSE 142 XV A LITTLE DEBIT IN YOUR TONNEAU 153 XVI A ROMANCE IN ENCYCLOPBDIA LAND 161 XVII THE PASSING OF THE ORTHODOX PARADOX . . .. . . 168 ix X CONTENTS CHAPTEE PAGE EXPLAINED . . . 175 XVIII SHAKESPEARE XIX Tm SCIENT

IFIC SCENARIO . . . 180 XX Tm MOST POPUL BOOK OF TBE MONTH ., . . . . . 187 AFTERNOON . . . . 193 XXI CHRISTMAS XXII m, VERNAL EQUINOX . . . 200 TABLOID EDITI

v11 absolute進入發燒排行的影片

愷他命濫用對尿液蛋白體及膀胱細胞自噬作用的影響

為了解決v11 absolute的問題，作者劉若娟 這樣論述：

Table of Contents指導教授推薦書………………………………………………………….口試委員審定書…………………………………………………………致謝…………………………………………………………….....………iii中文摘要……………………………………………………….....………ivAbstract………………………………………………………..…..…..….viTable of Contents ……………………………………….…………...…viiiList of Figures ......………………………………………….……....……xiiList of Tables.

.…………………………………………………..…...…..xivAbbreviation………………………………………………….….…..….xviChapter I Introduction ……………………………………….…..….…..11.1 The controlled substance Ketamine and diseases of bladder…....……..11.2 Ketamine and its metabolites in urine…………………………..…..….21.3 Exploration the molecular mechanism of bladd

er dysfunction which induced by chronic ketamine abuse …………………………….…..…..31.4 The possible mechanisms of ketamine -induced ulcerative cystitis ..…41.5 Ketamine induced autophagy process........................………………..…51.6 Specific aims of this thesis………………………………………....…..6Chapter II Material and Me

thods………………………………....…….72.1 Study subjects and sample collection…………………………….....….72.2 LC-SRM-MS analysis of ketamine and metabolites in urine sample.....82.3 Concentration and desalting of urine samples………………………....92.4 Tryptic digestion of urinary proteins and iTRAQ reagent labeling........112.5

Two-dimensional LC-MS/MS analysis ………………………….......112.6 Sequence database search and quantitative data analysis………....….122.7 Bioinformatics and network analysis…………………………....……132.8 Western blot analysis………………………………………….....……142.9 Quantification of APOA1 and SAA4 by ELISA …………...…..….152.10 Stat

istical analysis…………………………………………….…......152.11 Chemicals and Reagents……………………………………….…....162.12 Cell Culture…………………………………………………….…....162.13 Drug Treatments and Preparation of Cell Extracts…………….…....172.14 Measurement of Reactive Oxygen Species (ROS) production…..….182.15 iTRAQ Reagent Labeling a

nd Fractionation by SCX for cell extractprotein …………………………………………………………....… 182.16 LC- MS/MS Analysis by LTQ-Orbitrap PQD for cell extractprotein ………………………………………………….……..……..192.17 Sequence Database Searching and Quantitative Data Analysis for cell extract protein …………………………………………………........ 202.18

Immunofluorescence cell images………………………….………...222.19 Animals and Ketamine Administration ………………….……….....232.20 Lysosome Isolation ……………………………………….………....232.21 Ketamine quantitation by LC-SRM-MS analysis in intracellularlysosome samples …………………………………….......................24Chapter III Results ……

………………………………………...........…27Part I. Association of urinary ketamine and APOA1 levels with bladder dysfunction in ketamine abusers revealed via proteomics and targeted metabolite analyses …………………………………….........………....... 273.1 Study population and experimental design................................

............273.2 Clinicopathological features of KA .......................................................283.3 Urinary ketamine, norketamine and dehydronorketamine contents in KA.......................................................................................................293.4 Quantitati

ve proteome profiling of urine samples from KA and healthyControls.................................................................................................303.5 Biological process network analysis of differentially regulated urinaryproteins in KA..............................................

.........................................313.6 Verification of differentially regulated proteins in urine samples ofhealthy controls and KA........................................................................333.7 Associations between urinary levels of ketamine and its metabolitesand clinicopatho

logical features............................................................343.8 Associations between urinary APOA1/SAA4 levels and clinicopathological features of KA......................................................35Part II. A controlled substance and anesthesia medicine ketamine induced a

utophagosome accumulation in bladder epithelial cells.........................363.9 Quantitative profiling of the proteome changes of bladder Apex cellstreated with ketamine at different time intervals....................................363.10 Ketamine induces puncta and cellular degradative compart

ments formation in Apex bladder epithelial cells...................................................383.11 The autophagy marker protein-MAP1LC3B expression in vivo.........393.12 Ketamine inhibits autophagic flux.......................................................403.13 Ketamine inhibits lysosome

acidification to block the formation ofautolyosome and autophagic degradation............................................413.14 Part of ketamine accumulation in intercellular lysosome ..................443.15 The ROS scavengers, Trolox, reduces the autophagosome. accumulation which induced by ket

amine ..........................................46Chapter IV Discussion .............................................................................484.1 Ketamine, norketamine and dehydronorketamine in KA urine............484.2 Ketamine and overactive bladder syndrome ..............................

..........494.3 Variation analysis of urine protein with ketamine abuse.......................504.4 The upregulated candidate protein- APOA1 in KA urine ......................514.5 Association of urinary ketamine and APOA1 levels with bladder dysfunction in ketamine abusers .........................

........................................524.6 Ketamine induced autophagy process change in bladder cells .............534.7 Ketamine accumulation in cell organelles.............................................544.8 ROS reduction may decrease autophagosome accumulation to reducecellular dysfunction .

.............................................................................554.9 Conclusion ............................................................................................56References..................................................................................................58Fi

gures and Tables.....................................................................................66List of FiguresFigure 1. Workflow for exploring urine proteome alteration in KA and the association between urine protein levels, contents of ketamine and its metabolites and clinicopathlogical fea

tures of KA......................................66Figure 2. Response curves for quantification of ketamine, norketamine and dehydronorketamine generated via LC-SRM-MS analysis.........................67Figure 3. Correlations of urinary levels of urinary levels of ketamine(K), norketamine (NK) and d

ehydronorketamine (DHNK) in ketamine abuser68Figure 4. Quantitative comparison of urine proteome profiles between KA and HC for both genders.............................................................................69Figure 5. Biological process network analysis of differentially regulated urine p

roteins in KA...................................................................................70Figure 6. Validation of differentially regulated proteins in urine samples of HC and KA groups......................................................................................72Figure 7. Detection

of 5 candidate proteins in ketamine abuser’s urine samples by Western blotting........................................................................74Figure 8. The IC50 of ketamine and norketamine in inhibiting the viability of normal bladder epithelial cells.......................................

.........................75Figure 9. Identification of differentially expressed proteins in ketamine treated bladder cells.....................................................................................76Figure 10. Ketamine modulates the autophagy process in human normal bladder Apex cells.....

..................................................................................79Figure 11. Effects of long-term ketamine treatment on the morphology and MAP1LC3B expression of bladder tissues in mouse model and patients with ketamine abuse. .......................................................

...................................82Figure 12. Ketamine inhibits autophagic flux in bladder epithelial cells... 86Figure 13. Ketamine inhibits lysosome acidification to block the formation of autolysosome and autophagic degradation..............................................89Figure 14. Detection

of ketamine in cellular lysosomes by using immunoprecipitation and UPLC-LC- SRM-MS.........................................92Figure 15. Detection of ketamine in cellular lysosomes by using sucrose density gradient centrifugation and UPLC-LC-SRM-MS...........................95Figure 16. Effects of T

rolox (a ROS scavenger) on the ketamine-induced oxidative stress in bladder epithelial cells...................................................98Figure 17. Ketamine induces ROS production and autophagosome accumulation in bladder epithelial cells.....................................................1

02List of TablesTable 1. Demographic characteristics of enrolled subjects.......................103Table 2. Clinical information on ketamine abusers enrolled in this study.104Table 3. Clinicopathological features of ketamine abusers enrolled in this study.............................................

.............................................................105Table 4. LC-SRM-MS analysis of levels of ketamine (K), norketamine (NK) and dehydronorketamine (DHNK) in urine samples of ketamine users....106Table 5. Protein identification and quantification results from urine samples of ketamine abu

sers and healthy controls via iTRAQ................................107Table 6. List of proteins quantified in urine samples of ketamine abusers and healthy controls.........................................................................................108Table 7. List of the upregulated proteins in

urine samples from ketamine abusers.......................................................................................................109Table 8. List of the downregulated proteins in urine samples from ketamine abusers............................................................................

...........................111Table 9. GO process network of enriched urine proteins in ketamine abusers of both genders generated using DAVID bioinformatics resources...........113Table 10. KEGG pathway of enriched urine proteins in ketamine abusers of both genders generated using DAVID bioinformat

ics resources...............114Table 11. List of fibrosis-related proteins among the differentially expressed urinary proteins of the KA group...............................................................115Table12. Associations between urinary contents of ketamine and metabolites and clinicopathol

ogical features of KA..................................117Table 13. Correlations among clinicopathological features, APOA1 and SAA4 urine levels in 56 KA......................................................................118Table 14. Up- and down-regulated proteins in ketamine-treated primary blad

der Apex cells.....................................................................................119Table 15. List of proteins quantified in Apex cells treated with ketamine for different time periods.................................................................................120Table 16. List

of the upregulated proteins in Apex cells treated with ketamine for different time periods............................................................121Table 17. List of the downregulated proteins in Apex cells treated with ketamine for different time periods.......................................

....................124Table 18. GO network analysis of the 210 differentially regulated proteins in Apex cells treated with ketamine for 24 hours using DAVID bioinformatics resources...........................................................................127Table 19. KEGG pathway analysis of the

210 differentially regulated proteins in Apex cells treated with ketamine for 24 hours using DAVID bioinformatics resources...........................................................................128Table 20. A partial list of well-known autophagy-related proteins detected as the up-regulated prot

eins in ketamine-treated Apex cells......................129