本文選題：輸血 + 新鮮冰凍血漿��； 參考：《吉林大學》2017年碩士論文

【摘要】：目的:血漿占血液成分的50%,血漿輸注是臨床治療的重要組成部分。臨床主要應用亞甲藍病毒滅活血漿和新鮮冰凍血漿,以往涉及兩種血漿的對比研究多注重于分析血漿凝血因子、血漿蛋白含量及相應臨床生化指標的改變,但對這兩種血漿中特異性差異蛋白的研究甚少。本課題應用比較蛋白質組學的研究方法分析亞甲藍病毒滅活血漿和新鮮冰凍血漿這兩種血漿中特異性差異蛋白的改變,為臨床醫(yī)生對需要輸血病人的治療提供實驗和理論依據,以減少血漿不合理輸注和輸注無效,特別是對一些特殊病例提供參考資料。方法:1.對臨床62例輸注不同血漿患者的輸血后凝血酶原時間和纖維蛋白原含量進行統(tǒng)計學分析。2.三氯醋酸(TCA)/丙酮沉淀法去除血漿中高豐度白蛋白。3.硫酸銨沉淀法去除血漿中高豐度白蛋白。4.SDS-PAGE分離鑒定去除血漿中高豐度白蛋白后的血漿蛋白。5.Clean-up去除血漿蛋白中干擾2DE的物質。6.SDS-PAGE和2-DE驗證clean-up是否有效去除血漿蛋白中干擾2DE的物質。7.2-DE分離病毒滅活血漿和新鮮冰凍血漿的血漿蛋白。8.采用質譜法分析鑒定兩種血漿中顯示差異明顯的蛋白點。9.重新收集兩種不同的血漿18-74例,進行Western blot和ELISA檢測驗證其中兩種差異明顯的蛋白質含量。結果:1.經統(tǒng)計分析臨床中應用兩種不同血漿后患者的化驗數據,結果提示兩種血漿的應用均具有一定的治療效果,但新鮮冰凍血漿(FFP)組患者的凝血酶原時間(PT)值優(yōu)于亞甲藍病毒滅活血漿(MB-FP)組(P0.05),有統(tǒng)計學意義,而兩種的纖維蛋白原含量差異(P0.05),無統(tǒng)計學意義。2.三氯醋酸/丙酮沉淀法去除高豐度白蛋白比飽和硫酸銨沉淀法去除高豐度白蛋白的效果好。3.兩種方法去除血漿中高豐度白蛋白后的血漿,再應用clean-up方法對去除血漿中的干擾2DE的物質效果不明顯。4.亞甲藍病毒滅活血漿與新鮮冰凍血漿的2DE圖譜分析顯示,兩種血漿出現一些差異蛋白質的表達。5.經過質譜分析及數據庫比對鑒定出8個差異蛋白質點(含6種蛋白質)發(fā)生了修飾作用,它們是:Complement C1r subcomponent,Inter-alpha-trypsin i nhibitor heavy chain H4,Keratin(type II cytoskeletal 1),Hemopexin,Fibrinoge n gamma chain,Transthyretin.我們選取了Hemopexin,Keratin(type II cytoskel etal1)進行了ELISA和Western blot驗證。6.對18例血漿的Western Blot檢測結果表明在新鮮冰凍血漿和病毒滅活血漿中均檢測到Keratin,type II cytoskeletal1,但其蛋白條帶的表觀差異并不明顯。7.對48例血漿的ELISA檢測結果表明,新鮮冰凍血漿中Hemopexin含量明顯高于亞甲藍光化學法病毒滅活的血漿。結論:臨床輸注新鮮冰凍血漿后患者的凝血酶原時間(PT)明顯優(yōu)于輸注亞甲藍病毒滅活血漿。比較蛋白組學的研究結果揭示,亞甲藍病毒滅活血漿的同時使血漿中一些特殊蛋白質如:Complement C1r subcomponent,Inter-alpha-trypsin,Inhibitor heavy chain H4,Keratin,type II cytoskeletal 1,Hemopexin,Fibrinogen gamma chain,Transthyretin發(fā)生了明顯的修飾作用,而這些修飾作用對蛋白質的功能有何影響、對血漿臨床應用的效果有何影響值得我們深入研究。
[Abstract]:Objective: plasma is 50% of the blood components. Plasma infusion is an important part of clinical treatment. Clinical application of methylene blue virus inactivated plasma and fresh frozen plasma. The comparative study of two kinds of plasma in the past mainly focuses on the analysis of plasma coagulation factor, plasma protein content and the changes of corresponding clinical biochemical indexes, but the two kinds of plasma plasma protein content and corresponding clinical biochemical indexes change. There are few studies on specific differential proteins in plasma. This subject uses comparative proteomics to analyze the changes in specific differential proteins in two kinds of plasma, methylene blue virus inactivated plasma and fresh frozen plasma, to provide experimental and theoretical basis for clinicians to treat patients in need of blood transfusion in order to reduce the unreasonable plasma levels. Ineffective infusion and infusion, especially for some special cases. Methods: 1. pairs of clinical 62 patients with different plasma plasma prothrombin time and fibrinogen content after transfusion were statistically analyzed by.2. three chloroacetic acid (TCA) / acetone precipitation method to remove the high abundance albumin.3. ammonium sulfate precipitation in plasma to remove plasma High abundance albumin.4.SDS-PAGE separation and identification of plasma protein.5.Clean-up after removal of high abundance albumin in plasma to remove the substance.6.SDS-PAGE and 2-DE that interfere with 2DE in plasma protein to verify whether clean-up effectively removes the substance that interferes with 2DE in plasma protein,.7.2-DE separation virus inactivating blood plasma and fresh frozen plasma protein.8. extraction Mass spectrometric analysis was used to identify two different plasma levels of protein point.9., which were significantly different in two kinds of plasma, and two different plasma levels were re collected, and Western blot and ELISA tests were used to verify the protein content. Results: 1. the data of two different blood plasma patients were statistically analyzed by statistical analysis. The results suggested two kinds of blood. The application of the plasma has a certain therapeutic effect, but the prothrombin time (PT) value of the fresh frozen plasma (FFP) group is better than the methylene inactivated plasma (MB-FP) group (P0.05), and the difference between the two kinds of fibrinogen (P0.05) is not statistically significant, and there is no statistically significant.2. three chloroacetic acid / acetone precipitation method to remove the high abundance albumin ratio. The effect of the removal of high abundances with saturated ammonium sulfate precipitation method is good.3. two methods to remove the plasma of high abundance albumin in plasma, and then the effect of clean-up method on removing the interference of 2DE in plasma is not obvious, and the analysis of 2DE Atlas of.4. methylene blue virus inactivated plasma and fresh frozen plasma shows that some kinds of plasma appear some of them. The expression of differential protein.5. identified 8 different protein points (including 6 proteins) through mass spectrometric analysis and database comparison. They are: Complement C1r subcomponent, Inter-alpha-trypsin I nhibitor heavy chain H4, Keratin (type 1). We selected Hemopexin, Keratin (type II cytoskel etal1) and ELISA and Western blot verification.6. for Western Blot test results of 18 cases of plasma. The results showed that the content of Hemopexin in fresh frozen plasma was significantly higher than that of methylene blue photochemical inactivated plasma. Conclusion: the prothrombin time (PT) of patients after infusion of fresh frozen plasma was significantly better than that of inactivated methylene blue virus inactivated plasma. Comparison of the results of proteomics revealed that methylene blue virus inactivated plasma. There are some special proteins in plasma such as Complement C1r subcomponent, Inter-alpha-trypsin, Inhibitor heavy chain H4, Keratin, type II cytoskeletal 1. What is the effect of the effect is worth studying in depth.

【學位授予單位】：吉林大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R457.1

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