新的植物毒素蒜头果蛋白的荧光光谱研究

蒜头果蛋白(Malanin)是从我国稀有植物蒜头果中分离纯化出的一种具有高细胞毒性的蛋白质。用荧光光谱法研究在温度、酸度、有机溶剂、表面活性剂、变性剂及荧光猝灭剂等不同条件对蒜头果蛋白溶液构象的变化。实验表明,Malanin在天然状态下荧光发射峰位于340 nm处,色氨酸(Trp)残基较大程度位于Malanin分子的疏水区。十二烷基硫酸钠、异硫氰酸胍、丙烯酰胺和碘化钾的加入均可使Malanin的分子构象发生变化,导致分子内Trp残基的荧光猝灭。异硫氰酸胍的加入使Trp残基的荧光发射峰位明显红移,表明位于Malanin分子较疏水环境内的Trp残基相对外露。     

Multiple-trap laser tweezers Raman spectroscopy for simultaneous monitoring of the biological dynamics of multiple individual cells

We report the development of a multiple-trap laser tweezers Raman spectroscopy (LTRS) array for simultaneously acquiring Raman spectra of individual cells in physiological environments. This LTRS-array technique was also combined with phase contrast and fluorescence microscopy, allowing measurement of Raman spectra, refractility, and fluorescence images of individual cells with a temporal resolution of ~5 s. As a demonstration, we used this technique to monitor multiple Bacillus cereus spores germinating in a nutrient medium for up to 90min and observed the kinetics of dipicolinic acid release and uptake of nucleic acid-binding stain molecules during spore germination.     

Structure and Anti-Tumor Activity of Bovine alpha-Lactalbumin after Binding Linoleic AcidSCIEI北大核心CSCD

The structure changes of alpha-lactalbumin after binding oleic acid and linoleic acid, including hydrophobic amino acids, hydrophobic regions, tertiary structure, secondary structure, was studied by intrinsic fluorescence, ANS-binding intrinsic fluorescence and circular dichroism spectrum, respectively. The anti-tumor activity of the lactalbumin-oleic acid complex and lactalbumin-linoleic acid complex was measured using the methylene blue method. It can be seen from the fluorescence spectra that a significant red-shift from 331.07 to 337.67 nm and 337.60 nm of alpha-lactalbumin occurred after binding of oleic acid and linoleic acid, respectively. Together with the ANS-binding spectra, which exhibited a blue-shift (from 516.20 to 514.10 nm and 508.50 nm, respectively) with an increased fluorescence intensity, it can be indicated that binding of oleic acid and linoleic acid lead to the exposure of hydrophobic amino acids and hydrophobic regions. Results of circular dichroism spectra indicated the partial loss of the tertiary structure, and an decrease of beta-turn and random coil, which turn to the p-sheet structure. Furthermore, the antitumor activity of the two complexes was verified on the three types of tumor cells. This study laid a theoretical basis for the development of anti-tumor drugs.     

丁香有效部位对LDL中脂类、蛋白氧化修饰及光谱学变化的抑制

LDL氧化修饰是致动脉粥样硬化的关键性因素。其中,在氧化修饰过程中LDL所含脂类、蛋白及其光谱学性质均会受到影响。前期研究结果发现丁香对LDL氧化修饰具有很强的抑制作用。但是,丁香是否是通过抑制脂类及蛋白氧化来实现,以及丁香是否能抑制LDL氧化修饰后光谱学性质的改变,这些问题目前均不清楚。据此,运用紫外吸收光谱、荧光光谱等方法研究了丁香有效部位(effective fraction from clove,EFC)对LDL氧化修饰的抑制效果。研究结果表明,EFC能有效延缓脂质氧化过程中共轭二烯(CD)增殖及推后其达到最大值,并抑制胆固醇降解;也可抑制蛋白组分apoB100中Trp荧光猝灭及Lys氧化修饰和荧光光谱变化;同时也能抑制LDL中脂质与蛋白过氧化产物-脂褐素的生成;此外,该部位对LDL 氧化修饰过程中紫外-可见光谱学特征的改变也有影响。论文将为丁香抗AS功能食品研发提供参考。     

Analyzing of LED-induced Blood Fluorescent Spectra

The visible fluorescence spectra of healthy mice blood cells have been measured by light emitting diode (LED) excitation at about 570 nm (Δλ1/2≈32 nm ) in vitro. It is found that the spectral profiles of mouse blood, erythrocyte and hemoglobin are similar. The physical mechanism for LED-induced blood cells fluorescence spectra is analyzed. The development of low intensity laser or light irradiating blood therapy in vivo and in vitro may be guided by the understanding of blood fluorescence characteristics.     

Analyzing of LED-induced Blood Fluorescent Spectra

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牛α-乳白蛋白-亚油酸复合物的结构及抗肿瘤活性

利用内源性荧光光谱、荧光探针(ANS)结合荧光光谱及圆二色谱,以乳白蛋白-油酸为参照,研究了乳白蛋白结合亚油酸后,疏水性氨基酸、疏水性区域、三级结构及二级结构的变化,并利用亚甲基蓝的方法评价了该复合物的抗肿瘤活性。荧光光谱结果显示,与乳白蛋白-油酸复合物类似,乳白蛋白结合亚油酸后,其内源性荧光光谱显著红移,从331.07 nm移至337.60 nm;外源性ANS结合光谱蓝移,从516.20 nm移至508.50 nm;且荧光强度增加,表明乳白蛋白结合亚油酸后同样出现了疏水性氨基酸及疏水性区域暴露的现象。圆二色谱结果表明,与乳白蛋白-油酸复合物类似,乳白蛋白结合亚油酸后,三级结构部分丧失,二级结构中β-转角及无规卷曲的含量显著降低,β-折叠含量增加。细胞实验证实了乳白蛋白-亚油酸复合物具有良好的抗肿瘤效果。该研究从复合物结构和功能的两方面,为新型抗肿瘤乳白蛋白-亚油酸复合物的开发提供了依据。     

小分子荧光探针的光谱

运用紫外-可见光谱、荧光光谱法研究了以二烯酮为荧光团的1,5-双(4-羟基)-1,4-戊二烯-3-酮探针的光学性质.一系列pH滴定实验表明探针的紫外吸收和荧光光谱随溶液的pH值的改变而变化.同时,探针紫外吸收光谱发生明显的红移,荧光光谱强度也随之变化,在碱性条件下,相对荧光强度浮动较大.解离常数pK<,α>为9.25,...     

癌细胞对血清光谱的影响

研究了在血清中加入不同剂量的癌细胞对血清光谱的影响。比较了两种癌细胞的光谱 ,并与正常血清的谱线做了比较。实验结果表明 :随着癌细胞个数的降低 ,吸收谱和荧光谱强度均呈增加趋势 ,但增加到一定程度时又开始下降 ;正常血清的谱线强度明显较强。结论 :荧光光谱表明癌细胞抑制了正常血清中某种基团的振动 ,吸收光谱则表明癌细胞抑制了酪氨酸对光的吸收。     

1,5-萘二胺衍生物的光谱分析及发光性能研究

合成了一种纯度较高的1,5-萘二胺衍生物(NPN),制备了NPN薄膜.利用紫外-可见吸收光谱和荧光发射光谱研究了NPN的发光行为,并结合电化学循环伏安法研究了其电子能级结构.结果表明,NPN的荧光光谱表现出明显的溶剂效应,认为其发生了从电子给体N原子到电子受体芳环之间的分子内电子转移,形成分子内电子转移络合物;从NPN薄膜与其溶液的吸收光谱峰值比较中看出吸收峰红移,认为薄膜中分子形成"J-聚集体";NPN的HOMO能级为-5.74 eV,光学禁带为2.79 eV;在365 nm紫外光的激发下,产生发光峰在448.6 nm附近、谱线带宽为72.6 nm的蓝光发射,发光亮度高,色纯度高.     

不同波长激发光对血清荧光光谱影响的实验研究

采用日本岛津荧光光度计RF5301,研究了血清的荧光光谱与激发光波长的关系。实验结果表明：在不同波长的紫外光激励下,血清产生的荧光光谱线型及峰值波长基本相同,与激励光波长无关,但荧光峰强度随激励光波长变化而变化。血清的荧光光谱有两个较强的荧光发射区,其中第一个发射区处于300～410 nm,第二个发射区处于410～530 nm。当激发光波长小于310 nm,荧光主要集中在第一发射区,荧光峰位于330和370 nm处,并产生竞争现象。当激发光波长大于250 nm时,只出现330 nm处的荧光峰,其最佳激励光波长为300 nm;当激发光波长大于320 nm,第一发射区的荧光变弱,在第二发射区的荧光变强,荧光峰位于452 nm。此研究为血液的光谱特性研究提供了实验依据,对光诱导荧光光谱诊断技术中激发光波长的选择具有一定的参考价值。     

荧光法研究木犀草素与人血清白蛋白的相互作用

用荧光光谱、同步荧光光谱和紫外吸收光谱方法,研究了木犀草素与人血清白蛋白（HSA）的相互作用。研究表明木犀草素对HSA有较强的荧光猝灭作用,根据不同温度下木犀草素对HSA的荧光猝灭作用,利用Stern-Volmer方程处理实验数据,表明木犀草素对HSA的荧光猝灭作用属于静态猝灭。根据Fōrster非辐射能量转移理论计算了木犀草素与HSA间的结合常数和结合位点数,求得了木犀草素与HSA间的结合距离r。热力学数据表明二者主要靠疏水作用力结合。同时用同步荧光光谱探讨了木犀草素对HSA构象的影响。     

Fluorescence of a photosensitizer based on an indotricarbocyanine dye in photochemotherapy

We present the results for studies of the spectral luminescence properties of a symmetric indotricarbocyanine dye (PD1) in HeLa tumor cells and animal tissues in vivo during a photochemotherapy session and after the end of the session. We have established that when the dye is exposed to light in tumor tissues, changes occur in the position and half-width of the dye fluorescence spectra, while in a culture of HeLa cells its spectral characteristics are constant. Based on analysis of the effect of overlap between the absorption spectra of endogenous biomolecules and the fluorescence spectra of the dye plus comparison of the experimental data with numerical modeling results, we have concluded that the observed changes in the fluorescence spectra of PD1 in vivo are due to a change in the ratio of the different forms of hemoglobin in the tumor tissue. We have shown that the spectral characteristics of PD1, fluorescing in the near IR range, correlate with the depth of tumor tissue necrosis achieved on exposure to light. We have established that tumor tissue necrosis occurs down to a depth of 2 cm in the case of all strains studied: S-45, SM-1, and W-256, where as a result of exposure to light, we observe an increase in the half-width and a short-wavelength shift of the fluorescence spectrum of the dye PD1, and also the intensity of its fluorescence does not recover.     

Measurement of chlorophyll in water based on laser-induced fluorescence spectroscopy: Using spiral-wound unclad optical fiber system and partial least squares regression

The feasibility of laser-induced fluorescence (LIF) method for chlorophyll concentration determination in water was investigated in this study. The chlorophyll fluorescence spectra were received by a spiral-wound unclad optical fiber system. Baseline and smoothing correction methods were studied. Statistical equations were established between reference data and fluorescence spectra by linear regression and partial least squares (PLSs) regression. The PLS regression outperformed linear regression by higher correlation coefficient of 0.9996063 and root mean square error value of 0.0221807. Based on the results, it was concluded that laser-induced fluorescence method with a spiral-wound unclad optical fiber system was suitable for chlorophyll concentration determination in water, and PLS regression was a reliable method for fluorescence spectra analysis.     

芪盐LB多层膜的三维荧光谱分析

采用稳态、时间分辨荧光和三维荧光光谱技术研究了芪盐有机分子在溶液和芪盐/花生酸交替LB多层膜中的激发态的动力学特性.研究表明,芪盐分子在LB膜中形成H-聚集体,使得其荧光光谱发生蓝移.由于偶极间的相互作用使得聚集体的能级发生分裂,荧光衰减曲线可用双指数函数进行拟合.三维荧光谱发现随着衰减时间的推移,荧光峰逐渐向长波方向移动,不同波段具有不同的衰减驰豫时间.     

消癌平诱导人肺腺癌（ASTC-a-1）细胞内caspase-3活化的荧光光谱分析

采用CCK-8技术检测发现传统中药消癌平（XAP）对人肺腺癌（ASTC-a-1）细胞的增殖活性具有显著的抑制作用。利用共聚焦扫描荧光显微成像、荧光共振能量转移（FRET）及其受体光漂白技术证实了基于FRET技术构建的SCAT3质粒在ASTC-a-1细胞中的稳定表达。将消癌平加入细胞培养液中培育细胞,并在不同的时间检测活细胞内SCAT3的荧光发射光谱,从而监测细胞中caspase-3的活化状态。实验结果表明：(1)消癌平可以有效抑制人肺腺癌（ASTC-a-1）细胞的增殖活性并诱导细胞的死亡,消癌平对细胞的抑制作用具有剂量依赖性。(2)消癌平处理细胞72 h后,细胞内大量的SCAT3被切割,表明细胞内caspase-3的活化水平明显升高。(3)将含消癌平的细胞培养液与细胞共同培养24 h,然后采用没有消癌平的细胞培养液培养细胞48和72 h后,细胞内SCAT3的光谱没有明显改变,表明消癌平作用细胞24 h内没有显著激活细胞内的caspase-3。     

新型Eu~(3+)、Tb~(3+)掺杂配合物的合成与荧光性能研究

合成了稀土(Eu3+和Tb3+)与二苯甲酰甲烷(DBM)、2,2′-联吡啶(Dipy)的一系列稀土配合物EuxTb1-x(DBM)3Dipy。元素分析和红外分析确定了配合物的组成,荧光光谱研究了荧光性质。铽掺入配合物后,铽能极大地增强铕的特征荧光,铽对铕配合物的发光有协同作用。在该系列配合物中,不仅有机配体可以将吸收的能量传递给发光的铕离子使其发光,而且铽离子也可将其吸收的能量通过分子内能量传递给铕离子。     

S-构型转化对卵白蛋白微观结构的影响

采用圆二色谱(CD)、X射线衍射(XRD)、ANS荧光探针和紫外光谱(UV)研究了S-构型转化对卵白蛋白微观结构的影响.结果显示,不同诱导时间处理的卵白蛋白二级结构的α-螺旋,β-折叠,β-转角和无规卷曲之间相互转化,α-螺旋略有减少,β-折叠相应增加,分子有序性提高;S-构型转化后卵白蛋白晶体结构增加,72 h处理后...     

Absorption spectra and spectral-kinetic characteristics of the fluorescence of Sanguinarine in complexes with polyelectrolytes and DNA

The absorption spectra and stationary and time resolved fluorescence spectra of the isoquinoline alkaloid sanguinarine are studied in aqueous media and during interactions with synthetic polyelectrolytes (polystyrene sulfonate and polyallylamine) and a natural polyelectrolyte (DNA).     

3-羟基-2-萘甲酸-铬(Ⅲ)配合物与牛血清白蛋白作用的光谱研究

研究了3-羟基-2-萘甲酸铬(I)配合物[Cr(3-HNA)(en)2]Cl的紫外-可见吸收光谱和荧光光谱性质.在pH 7.4.0.05mol.L-1 Tris-HCl缓冲条件下通过荧光光谱技术,研究了BSA与[Cr(3-HNA)(en)2] 间的相互作用,测定了反应的结合常数与结合位点数.