(由吴明道教授讲解)

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1、Blood specimen preparationGlucose analysisLiver function testNeutral sugar and sialic acid determination(由吳明道教授講解)Blood sample collectionHeparinHeparin: Direct thrombin inhibitors Blood glucose determinationHormonal regulationInsulin: Release from b-cells islet of Langerhans of pancreas in response

2、to high blood glucoseIncrease uptake of glucoseIncrease synthesis of glycolytic enzymes, glucokinase, PFK, & PKInhibits gluconeogenesisGlucagon: -cells islets of Langerhans of pancreas in response to low blood glucoseStimulate liver (not muscle) to release glucoseInvolved in long term glucose mainte

3、nanceEpinephrine (adrenaline): released by adrenal glandsActive on liver and muscleStimulates glycogen breakdown and glycolysisThe typical human adult uses 160 g of glucose per day, with 120 g being used by the brain. The available reserve of glucose is adequate for about one day (about 20 g of circ

4、ulating glucose and 190 g of glycogen). After the reserve becomes depleted, how would a starved body obtain more glucose? In animals, many precursors lead to the synthesis of glucose (gluconeogenesis). In humans, the principal precursors are glycerol from triacylglycerols and glucogenic amino acids

5、from proteins. Blood glucose determinationGlucose oxidase2H2O + Glucose + O2 gluconic acid + H2O22H2O2 + 4-aminoantipyrine (AAP) 4H2O + Quinoneimine Dye (OD505, pink)peroxidase血糖的正常值血糖的正常值 正常人空腹時的血糖值為正常人空腹時的血糖值為 70-110 mg/dl 將空腹血糖異常值從110 mg/dL降到了100 mg/dL,可能使前驅前糖尿病的診断增加约20% 預料國內將多出近預料國內將多出近20萬名代謝症候群

6、病患萬名代謝症候群病患 2003年下修Liver function testGOT and GPT determination肝功能測試肝功能測試:決定你人生是決定你人生是黑黑白白或或彩彩色色Transaminase: GOT(AST), GPT(ALT)Having the function of transferring amino group of amino acids from alpha-amino acids to alpha-keto acids, therefore, named transaminase. GOT (Glutamyl oxaloacetic transami

7、nase, Aspartate aminotransferase, AST) L-Aspartic Acid + Ketoglutaric Acid Oxaloacetate + L-GlutamateGPT (Glutamyl pyruvic transaminase, Alanine aminotransferase, ALT) L-Alanine + Ketoglutaric acid Pyruvate + GlutamateOne of the thousands kinds of liver enzymes, and a kind of transferase. Storage mo

8、stly in liver etc. Low serum concentration. Large amount of transaminase is released into blood mostly on liver cell damages. -Thus, detection of serum level tells the existence of liver cell damage. Transaminase: GOT(AST), GPT(ALT)OrgansGOTGPTMyocardiumLiver KidneyPancreasSpleenLungRed blood cellSe

9、rum156,000142,000 99,000 28,000 14,00010,000300207,10044,0004,8002,0001,20072011016Concentration in organs and tissues (Karmen unit/1 gm wet weight) Chapter 16 TCA cycleMalate aspartate shuttleMETHOD PRINCIPLE In this method ALT catalyzes the reaction of L-alanine and - ketoglutaric acid to form pyr

10、uvate and glutamate under controlled conditions (37 C) and pH 7.4 + 0.05. The addition of acidic 2,4-dinitrophenylhydrazine stops the reaction and forms the 2,4-dinitrophenylhydrazone. So that it may be measured at 505 + 5 nm.測定血液中的GOT及GPT活性Reitman & Frankel 比色法 (酮酸-DNP呈色法)-Keto acidDNP (2,4-dinitro

11、phenylhydrazine) (金黃色)2,4-dinitrophenylhydrazone (紅棕色)RC COOHONH2NHNO2NO2RCCOOHNNO2NO2NH+OH-(Pyruvate or OXA)Aspartic acid or alanineTransaminase (GOT or GPT)GOT及GPT的正常值 Serum GOT(SF Units/ml)Serum GPT(SF Units/ml)正常值028021臨界值28402135S. F. Unit (Sigma-Frankel Unit):在25C,pH 7.5的狀態下,每分鐘能產生4.8210-4 m m

12、mol glutamate的酵素活性。I. Unit (International Unit):在特殊狀態下,每分鐘能轉變1 m mmol受質的酵素量。1 SF U/ml = 0.48 IU/L GOT及GPT的比較 CharacteristicGOTGPTPresent in tissues other than liverMore in heart than in liver; also in skeletal muscle, kidney, brainRelative low conc. in other tissueLocation in hepatocyte Mitochondria

13、 and cytoplasmCytoplasm onlyReference range in adult blood 10-40 IU/liter5-35 IU/literHalf-life in blood12-22 hr37-57 hrChange with acute inflammatory damageModerately sensitiveExtremely sensitiveChange with primary or secondary neoplasmSubstantial riseModerate or no riseChange with cirrhosisModerate riseMild or moderate riseChange with myocardial infarctSubstantial riseMild or moderate riseThank you(GOT) (GPT)

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