【作者】 兰秀风；

【导师】 倪晓武； 陆建； 骆晓森； 沈中华；

【作者基本信息】 南京理工大学 ， 光学工程， 2005， 博士

【摘要】 本文从理论和实验上研究了光诱导人全血、人脑硫氧还蛋白还原酶(TrxR)的荧光光谱及其特性,以及高胆固醇血症血清的光谱学检测机理与方法。 根据波长为407nm的LED光诱导的人与实验小白鼠的全血荧光光谱谱线形状、谱线随血细胞浓度的变化趋势以及荧光的发光机理相似的现象,首次提出了在进行光与人血液相互作用机理研究时,可研究光与小自鼠血液相互作用的光谱学特性、发光机理及其产生的生物效应的物理机制,为研究光与人血相互作用机理提供参考。 通过对波长为457nm的LED光、波长为457.9nm的Ar~+激光和波长为632.8nm的He-Ne激光激励下的全血荧光光谱进行比较研究发现:采用激光光源激发可以显现血细胞荧光光谱的部分精细结构,这说明血细胞中的荧光团对激发光波长具有一定的选择性,这种选择是由其自身的能级结构和状态所决定的。此外,He-Ne激光诱导的血液荧光光谱强度相对较弱,因此在ILLLIT临床应用中,He-Ne激光引起的光生物效应也较小。由于弱激光的生物效应具有双相性和可累积性等特点,故采用He-Ne激光进行血管内照射疗法,可较容易控制其照射剂量与作用效果,因而更为安全可靠。 在检测了波长为407nm的LED偏振光诱导人全血溶液静态荧光偏振光谱偏振度的基础上,研究了偏振度随浓度的变化规律,并采用能量转移理论分析了在不同荧光区、不同浓度下,血液中各荧光团之间的不同能量转换机制及偏振度的变化现象。 首次研究了正常人血清与高胆固醇血症血清的光谱学特性,结果表明正常人血清与高胆固醇血症血清的吸收光谱和荧光光谱的谱峰形状均存在较大的差异,且正常人血清的吸收率和荧光强度均较低,因此可以通过比较待测血清的吸收光谱和荧光光谱初步判定血清中胆固醇含量是否异常。 首次研究了波长为407nm的紫色光和波长为253.7nm的紫外光激励溶液状态下的人脑硫氧还蛋白还原酶(TrxR)分子的荧光光谱及其特性。理论与实验结果均表明TrxR的荧光光谱与TrxR分子的构象和空间结构密切相关。 本文的研究结果对研究光与血液相互作用的机理、借助血液及其组分的自体发光特性进行临床诊断和治疗、以及研究生物大分子的构象与结构特点提供实验和理论依据。更多还原

【Abstract】 Light induced fluorescence spectra and its characteristic of human blood and thioredoxin reducase (TrxR), and the spectroscopic detection of hypercholesterolemia serum are systemically investigated in experiment and theory.Based on the fact that the profile, the change tendency with the blood concentration change, and the excitation mechanism of fluorescence from human blood are similar to those from laboratory rats’ blood, it is first proposed that rat blood can be used as human blood to investagate the characteristic and the production mechanism of blood fluorescence spectra and the corresponding biological effects.Experimental results of LED light (λ=457 nm), Ar~+ laser (λ=457.9 nm) and He-Ne laser (λ=632.8 nm) induced whole blood fluorescence spectra indicate that using laser as excitation light can exhibit a part of fine structure of blood cell spectra, which suggests that fluorophores have a selective absorption to the wavelength of the exciting light and this selectivity are dependent on the energy level structure and the state of the fluorophores. In addition, the fluorescence spectra of blood excited by He-Ne laser is much lower than the other two. As a result, the photobiological effect of He-Ne laser is smaller. Since the biological effect of lower power laser is bi-directional and cumulative, it is easy to control the irradiation doze and the effectiveness if He-Ne laser is used in clinical therapy of the intravascular low-level laser irradiation (ILLLI).The fluorescence polarization of whole blood induced by LED polarized light at 407 nm and its variation rules with the blood concentration are investigated by experiment. Based on the theory of energy transfer, the energy transfer mechanism in different fluorescence areas, under different concentrations of whole blood as well as the variation mechanism of polarized degree are discussed.The spectroscopy characteristic of normal human serum and the hypercholesterolemia serum indicate that both the absorption spectra and the fluorescence spectra of hypercholesterolemia serum are different from those of normal human serum. Thus, the abnormal content of cholesterol in serum can be judged by comparing the absorption and fluorescence spectra.The fluorescence spectra of TrxR solution from human brain excitated by violet LED light at 407 nm and Ultra-violet light at 253.7 nm are investigated for the first time. The更多还原