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  本文關(guān)鍵詞：基于漫反射光譜法的皮膚膽固醇檢測技術(shù)研究　出處：《中國科學(xué)技術(shù)大學(xué)》2017年博士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 皮膚膽固醇 動脈粥樣硬化性心血管疾病 漫反射光譜 漫反射吸光度

【摘要】：動脈粥樣硬化性心血管疾病(atherosclerotic cardiovascular diseases,ASCVD)是危害人類健康的重大疾病之一,其診斷與預(yù)防已經(jīng)逐漸引起社會的關(guān)注。皮膚膽固醇與ASCVD密切相關(guān),成為ASCVD的新型生物標(biāo)志物。采用漫反射光譜法測量受試者皮膚膽固醇水平,進(jìn)而實現(xiàn)ASCVD的早期篩查和風(fēng)險評估,具備無創(chuàng)、便捷、實時和原位等顯著優(yōu)點,有較好的發(fā)展前景。本文重點研究基于漫反射光譜法的皮膚膽固醇無創(chuàng)檢測技術(shù),包括研制皮膚膽固醇特異性識別和指示所需的試劑,設(shè)計集成化的皮膚膽固醇光譜檢測系統(tǒng)。并開展實驗研究,通過測量離體豬皮膽固醇和在體人體皮膚膽固醇來驗證該方法和系統(tǒng)的可行性。本文的主要內(nèi)容和取得的成果有:1、皮膚膽固醇特異性識別和指示所需的試劑研制以及光譜檢測系統(tǒng)設(shè)計。分為兩個部分:第一部分為基于"三滴法"皮膚膽固醇特異性識別和指示所需試劑的設(shè)計和研制,包括檢測試劑"A-C-B"共聚物溶液和供氫體底物溶液。并通過檢測離體豬皮內(nèi)的膽固醇驗證了試劑的有效性。第二部分為光譜檢測系統(tǒng)的設(shè)計,分為硬件設(shè)計部分和軟件設(shè)計部分。硬件部分包括光源單元、光學(xué)傳輸單元、信號探測單元、試劑反應(yīng)腔單元等。軟件部分編寫了用于皮膚膽固醇光譜測量的控制分析軟件。除此之外,還進(jìn)行了殼體設(shè)計、附件設(shè)計等,最后完成整套系統(tǒng)的設(shè)計和研制。2、皮膚膽固醇光譜檢測系統(tǒng)性能測試,測量影響因素研究和校正。首先,對設(shè)計的皮膚膽固醇光譜檢測系統(tǒng)的性能進(jìn)行測試,包括光源、傳輸光路及微型光纖光譜儀。采用標(biāo)準(zhǔn)漫反射板和與待測對象相似形態(tài)的去離子水驗證系統(tǒng)的重復(fù)性。其次,對影響系統(tǒng)測量的因素進(jìn)行了研究和優(yōu)化,包括系統(tǒng)與組織接觸壓力的影響,辣根過氧化物酶(Horseradish Peroxidase,HRP)的活性,系統(tǒng)漂移和皮膚背景等。第三,采用研制的皮膚膽固醇光譜檢測系統(tǒng),采集漫反射光譜,求得漫反射吸光度A的曲線,確定特征波長和特征波段。最后,根據(jù)膽固醇與檢測試劑"A-C-B"共聚物溶液以等分子比例結(jié)合的特性,采用梯度濃度的檢測試劑模擬梯度濃度的皮膚膽固醇,計算漫反射吸光度A并與"A-C-B"共聚物溶液濃度進(jìn)行相關(guān)性分析。結(jié)果表明,特征波長652 nm處的漫反射吸光度A和特征波段620-650 nm內(nèi)的積分漫反射吸光度A與" A-C-B"共聚物溶液濃度成線性增長和顯著正相關(guān)。3、采用皮膚膽固醇光譜檢測系統(tǒng)實現(xiàn)離體豬皮膽固醇的測量。首先采用萃取法獲得含有梯度濃度膽固醇的離體豬皮樣本。確定了萃取的步驟和條件。然后采用光譜檢測系統(tǒng)檢測萃取后的樣本內(nèi)的膽固醇,并與萃取時間進(jìn)行線性擬合分析。結(jié)果表明,采用特征波長652nm處的漫反射吸光度A和特征波段620-650 nm的積分漫反射吸光度A均可以區(qū)分和相對定量樣本內(nèi)的梯度濃度膽固醇,并與萃取時間成線性相關(guān)。4、采用皮膚膽固醇光譜檢測系統(tǒng)實現(xiàn)人體在體皮膚膽固醇無創(chuàng)測量。征集受試者31名,采用設(shè)計的光譜檢測系統(tǒng)采集受試者在體皮膚膽固醇,并獲取受試者血液膽固醇。通過線性擬合和相關(guān)性分析受試者皮膚膽固醇與血液膽固醇中總膽固醇(total cholesterol,TC)、甘油三酯(triglyceride,TG)、高密度脂蛋白膽固醇(high density lipoprotein cholesterol,HDL-C)、低密度脂蛋白膽固醇(low density lipoprotein cholesterol,LDL-C)和極低密度脂蛋白膽固醇(very low density lipoprotein cholesterol,VLDL-C)5項指標(biāo)和由該5項指標(biāo)衍生出的其它ASCVD風(fēng)險指標(biāo)的關(guān)系。結(jié)果顯示:皮膚膽固醇與TC和LDL-C的變化趨勢一致,并和二者顯著正相關(guān),控制年齡、性別和BMI等因素后,偏相關(guān)系數(shù)r分別為 0.813(P.01和 0.759(P＜0.001);皮膚膽固醇與 TG、HDL-C 和 VLDL-C 3項指標(biāo)無顯著性關(guān)系;皮膚膽固醇與非HDL-C、TC/HDL-C、LDL-C/HDL-C和AI四項指標(biāo)之間均成線性關(guān)系,并成顯著正相關(guān),偏相關(guān)系數(shù)r分別為0.769(P0.01)、0.674(P0.01)、0.746(P0.01)和 0.674(P0.01);將皮膚膽固醇與任意兩項參考指標(biāo)進(jìn)行組合分析,隨著參考指標(biāo)的增長,皮膚膽固醇也呈增長趨勢。最后,基于上述參考指標(biāo)在ASCVD風(fēng)險評估中的參考標(biāo)準(zhǔn)將受試者劃分為正常組、低風(fēng)險組和高風(fēng)險組,3組受試者之間的皮膚膽固醇具有顯著性差異(單因素方差分析F值均大于1,P0.01),表明采用皮膚膽固醇能夠?qū)崿F(xiàn)不同ASCVD風(fēng)險受試者的區(qū)分。綜合上述研究表明,采用漫反射光譜法無創(chuàng)檢測皮膚膽固醇將有助于ASCVD早期篩查和風(fēng)險評估,對于該類疾病的防控具有一定的意義。
[Abstract]:Atherosclerotic cardiovascular disease (atherosclerotic cardiovascular, diseases, ASCVD) is one of the major diseases that endanger human health, its diagnosis and prevention has gradually attracted the attention of the society. The skin cholesterol is closely related with ASCVD, ASCVD has become a new biological marker. Measuring subjects skin cholesterol levels by using diffuse reflectance spectroscopy, and screening and risk assessment of early ASCVD, with noninvasive, convenient, real-time and in situ and other significant advantages, it has good prospects for development. This article focuses on the study of skin cholesterol diffuse reflectance spectroscopy noninvasive detection technology, including the development of skin cholesterol reagent specific recognition and instructions required for the skin cholesterol spectrum detection system integrated design and the experimental study was carried out by measuring the in vitro and in vivo human skin skin cholesterol cholesterol to verify the method and system The feasibility. The main contents of this paper are as follows: 1. Design and development of reagent spectrum detection system of skin cholesterol specific recognition and instructions required. Divided into two parts: the first part is based on the "three drops" skin cholesterol specific recognition and instructions required to test the design and development of agent, including kit "A-C-B" copolymer solution and hydrogen donor substrate solution. And through the detection of cholesterol in vitro pig validated reagents. The second part is the design of spectrum detection system, divided into hardware design and software design. The hardware part includes a light source unit, optical transmission unit, a signal detection unit, reagent the reaction cavity unit. The software is used to control skin cholesterol spectrum analysis software. In addition, also carried out the shell design, accessories design, finally completed the whole design of system With the development of.2, skin cholesterol spectrum detection system performance test, and the correction factor measurement. Firstly, the performance of skin cholesterol spectrum detection system design for testing, including the light source, optical transmission and micro fiber spectrometer. The repeatability standard diffuser and deionized water with the measured object similarity form verification system. Secondly, the research and optimization of factors affecting the measurement of the influence, including system and organization of the contact pressure, horseradish peroxidase (Horseradish Peroxidase, HRP) activity, system drift and skin background. Third, the skin cholesterol spectrum detection system development, acquisition by diffuse reflectance spectroscopy, diffuse reflectance the absorbance of A curve, determine the characteristics of wavelength and spectral characteristics. Finally, according to the characteristics of cholesterol and the kit "A-C-B" copolymer solution with molecular ratio with the harvest Simulation of skin cholesterol concentration gradient detection reagent concentration gradient calculation, diffuse reflectance absorbance A and analyzed the correlation with the concentration of A-C-B copolymer solution. The results show that the integral features of diffuse diffuse reflection at the wavelength of 652 nm A and 620-650 nm absorbance characteristic band in reflection absorbance A and "A-C-B" copolymer concentration linear growth and significant positive correlation with.3, skin cholesterol spectrum detection system achieve measurement of body cholesterol. The pigskin extraction method with gradient concentration of cholesterol in vitro. The process and conditions of pigskin sample extraction is determined. Then the spectrum detection system after extraction of cholesterol in the sample, and the linear regression analysis and the extraction time. The results show that the integral absorbance of A using diffuse reflectance characteristics of diffuse reflectance at a wavelength of 652nm absorbance of A and 620-650 nm are the characteristic bands To distinguish between the concentration gradient and relative quantitative cholesterol within the sample, and a linear relationship between the.4 and the extraction time, the skin cholesterol spectrum detection system to achieve human skin in vivo noninvasive measurement of cholesterol. For 31 participants, the system design of the spectra of subjects in skin cholesterol, and obtain the subjects blood cholesterol. Through the analysis of linear fitting and correlation between the subjects of skin cholesterol and blood cholesterol and total cholesterol (total cholesterol TC), triglyceride (triglyceride, TG), high density lipoprotein cholesterol (high density lipoprotein cholesterol, HDL-C), low density lipoprotein cholesterol (low density lipoprotein cholesterol, LDL-C) and very low density lipoprotein cholesterol (very low density lipoprotein cholesterol, VLDL-C) between the 5 indexes and derived from the 5 indicators of the ASCVD risk index. The results showed that the change trend of skin cholesterol and TC and LDL-C, significant positive correlation, and the two control factors such as age, gender and BMI, the partial correlation coefficient r was 0.813 (P.01 and 0.759 (P < 0.001); skin cholesterol and TG, there was no significant relationship between the 3 indexes and HDL-C VLDL-C; skin cholesterol and non HDL-C, TC/HDL-C, as are the linear relationship between the four indexes of LDL-C/HDL-C and AI, and a significant positive correlation, partial correlation coefficient r was 0.769 (P0.01), 0.674 (P0.01), 0.746 (P0.01) and 0.674 (P0.01); the two reference index of skin cholesterol with any combination of analysis, with the reference index of the growth of skin cholesterol showed a growth trend. Finally, the reference standard reference index in ASCVD risk evaluation based on the subjects were divided into normal group, low risk group and high risk group, the skin cholesterol between the 3 groups of subjects with significant Difference (single factor analysis of variance F-measure was greater than 1, P0.01), show that the skin cholesterol can distinguish different ASCVD risk subjects. The research shows that using diffuse reflectance spectroscopy in noninvasive detection of skin cholesterol will help ASCVD early screening and risk assessment, and has a certain significance for prevention and control this kind of disease.

【學(xué)位授予單位】：中國科學(xué)技術(shù)大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2017
【分類號】：R54;O657.3

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