用Fura-2测定人外周血中性粒细胞内游离钙浓度

本文用 2 - [6-双乙酸基 - 5 - ( 2 -双乙酸基 ) - 5 -甲苯氧乙氧基 ]- 2 -苯骈呋喃基 - 5 -恶唑羧酸五钾盐 ( Fura- 2 )建立了测定人外周血中性粒细胞内游离钙浓度的方法 ,并对一些测试条件进行了探讨。测定标本内中性粒细胞数以大约 10 6个 /m L为宜 ,在离心分离出中性粒细胞悬液时 ,标本制备好需静置 0 .5 h后才能上机测试 ,否则振荡形成的剪切应力将增大细胞内游离钙的浓度 ,p H偏离 7.4和受损细胞会极大增加细胞内游离钙的浓度。结果表明 ,正常人外周血中性粒细胞游离钙浓度为 3.95± 0 .76μg/L。对中性粒细胞内游离钙的测定有助于了解其功能状态。     

Studies on Calcium Release and H2O2 Level Produced by the Elicitor Induced Plant Cell by Fluorescence Probing

Using fluorescence probing technology, we studied the mechanism and interrelations of calcium release and H₂O₂ production in situ in living tissues of tobacco and cotton plants which were induced by pathogen elicitor, salicylic acid (SA) and pectinase respectively. Results showed that (1) pathogen elicitors could induced H₂O₂ response in epidermis cells regardless of environmental calcium, but in mesophyll protoplast, H₂O₂ response could only be induced at calcium condition. Similarly, SA and pectinase induced H₂O₂ response could only be observed at calcium condition; (2) pathogen elicitors could induce calcium response in both epidermis cells and protoplasts regardless of environmental calcium, while calcium response couldn’t be induced at non-calcium condition by SA and pectinase; (3) H₂O₂ response and calcium response in protoplast were faster than that in the whole cell. These results indicated that pathogen elicitors can induce the release of cell wall calcium and the cell wall calcium release is independent to pectinase. And it is concluded that free calcium influx is necessary for the oxidative burst and cell wall calcium has an irreplaceable role in defense signal transduction.     

Nuclear Magnetic Ions of Magnesium,Calcium, and Zinc as a Powerful and Universal Means for Killing Cancer Cells

Magnetic isotope effect controls enzymatic DNA synthesis and strongly, by 2–3 times, suppresses catalytic activity of polymerases and increases even more strongly, by 20–50 times, the mortality of cancer cells. Catalyzing ions ²⁵Mg²⁺, ⁴³Ca²⁺, and ⁶⁷Zn²⁺ carrying magnetic nuclei are shown to efficiently kill cancer cells. The advantage of these ions for practical medicine is that being injected in blood they are captured selectively and almost exclusively by cancer cells inducing their death. The healthy cells capture these ions much less efficiently (perhaps due to the lower penetrability of their membranes) and are not vulnerable to these ions in comparison with cancer cells. Of course, penetrability of cells is identical for magnetic and nonmagnetic ions, but only the former kill cancer cells.     

利用光诱导延迟荧光的植物盐胁迫生理监测

利用自制的便携式延迟荧光(Delayed Fluorescence,DF)探测系统,以早熟18号大豆品种为实验材料,研究了盐胁迫对植物叶绿体DF特性的影响.结果发现在单纯的盐胁迫和盐胁迫伤害程度被调控缓解时,绿色植物叶片光诱导DF强度的变化都能很好地反应植物叶片光合性能的变化.因此,可以用DF强度的变化来表征盐胁迫对植物的伤害程度.     

Fluorescence Polarization as a Functional Parameter in Monitoring Living Cells: Theory and Practice

The use of fluorescence polarization as a functional parameter in monitoring cellular activation calls for the reliable and accurate measurement of the fluorescence intensity and polarization (FI and FP) of microscopic objects. The relevant experimental parameters that enter such measurements are thoroughly discussed. The possibility of executing FP measurements properly by flow-through systems is compared with that of static cytometry. Remarks on the effects of high-power excitation on markers and cells conclude the paper.     

X射线荧光光谱法测定溴化锌溶液中溴化锌的含量

采用X射线荧光光谱法,以光谱纯溴化锌粉末配制系列标准点,建立校准曲线,测定溴化锌溶液中溴化锌的含量。对激发电压、电流及扫描角度等影响因素进行了实验研究。方法准确可靠、快速,相对标准偏差优于(RSD,n=8)1%,锌的回收率在108.0%—115.3%之间,溴的回收率在105.8%—114.0%之间。     

钙(Ⅱ)增敏秦皮乙素-CTMAB体系荧光特性的研究

研究了钙 ( ) -秦皮乙素 - CTMAB体系的荧光性质 ,发现 Ca2 +对秦皮乙素 - CTMAB有较强的增敏作用 ,建立了荧光光谱法测定痕量秦皮乙素的新方法。其线性范围为 2 .38× 10 -8— 2 .38× 10 -6mol· L-1;检出限为 1.2× 10 -8mol·L-1;回收率为 91.39%— 10 2 .31% ;相对标准偏差为 3.6 %。     

Probing the Interaction of Trans-resveratrol with Bovine Serum Albumin: A Fluorescence Quenching Study with Tachiya Model

The interaction of trans-resveratrol (TRES) and bovine serum albumin (BSA) was investigated using fluorescence spectroscopy (FS) with Tachiya model. The binding number maximum of TRES was determined to be 8.86 at 293.15 K, 23.42 at 303.15 K and 33.94 at 313.15 K and the binding mechanism analyzed in detail. The apparent binding constants (K (a)) between TRES and BSA were 5.02 x 10(4) (293.15 K), 8.89 x 10(4) (303.15 K) and 1.60 x 10(5) L mol(-1) (313.15 K), and the binding distances (r) between TRES and BSA were 2.44, 3.01, and 3.38 nm at 293.15, 303.15, and 313.15 K, respectively. The addition of TRES to BSA solution leads to the enhancement in RLS intensity, exhibiting the formation of the aggregate in solution. The negative entropy change and enthalpy change indicated that the interaction of TRES and BSA was driven mainly by van der Waals interactions and hydrogen bonds. The process of binding was a spontaneous process in which Gibbs free energy change was negative.     

Improved Voltage and Fill Factor by Using Zinc Oxide Thin Film as a Barrier Layer in Dye-Sensitized Solar Cells

A series of dye-sensitized solar cells based on ZnO-modified TiO2 nano-porous films have been prepared. The current-voltage characteristics of the cells show that the ZnO-modification can improve the open-circuit voltage and the fill factor but can decrease the short-circuit current. Dark current and transient photovoltage measurements are used to study the back reaction. It is indicated that the recombination process is suppressed by blocking the hole transporting from the nano-porous TiO2 since the surface of the semiconductor is almost fully covered with ZnO as a barrier layer.     

探针3-(2-氰基乙基)胞嘧啶同步荧光法测定血清中蛋白质

在模拟人体生理条件下,基于3-(2-氰基乙基)胞嘧啶(CECT)与人血清白蛋白(HSA)和牛血清白蛋白(BSA)之间的相互作用,以CECT为分子光谱探针研究了CECT-蛋白质体系的同步荧光光谱特征。同步荧光光谱特征及强度与Δλ值、反应介质、反应温度等因素有关。在此基础上,建立了以CECT为分子光谱探针定量测定血清样品中蛋白质含量的新方法。在最佳实验条件下,CECT-HSA和CECT-BSA体系的同步荧光强度分别在0～441.4和0～351.0 μg·mL-1的浓度范围内与蛋白质浓度呈现良好的线性关系, 检测限分别为0.023和0.035 μg·mL-1,相对标准偏差(RSD)1.2%～3.3%, 加标回收率为97.2%～100.4%。该方法具有简单、快速、灵敏度较高、线性范围宽、精密度和回收率较好等优点。该法可直接用于血清样品中蛋白质总量的测定,结果令人满意。     

流动注射浊点萃取结合荧光光谱法测定水样中的痕量锌

锌离子与8-羟基喹啉混合后与Triton X-114合并,在线生成的锌配合物被表面活性剂Triton X-114捕集保留到脱脂棉微型柱。乙醇洗脱分析物,荧光光谱法检测洗脱液中的痕量锌。考察了影响流动注射浊点萃取体系的各种因素,包括pH、配位剂浓度、表面活性剂浓度、样品富集和洗脱流速等。测定锌的线性范围10—200μg/L,检出限(3σ)为2.0μg/L。方法应用于饮用水和环境水样测定,加标回收率90.4%—107.8%。     

2-NBDG Fluorescence Imaging of Hypermetabolic Circulating Tumor Cells in Mouse Xenograft model of Breast Cancer

Objectives

To determine use of 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-D-glucose (2-NBDG) as a tracer for detection of hypermetabolic circulating tumor cells (CTC) by fluorescence imaging.

Procedures

Human breast cancer cells were implanted in the mammary gland fat pad of athymic mice to establish orthotopic human breast cancer xenografts as a mouse model of circulating breast cancer cells. Near-infrared fluorescence imaging of the tumor-bearing mice injected with 2-DeoxyGlucosone 750 (2-DG 750) was conducted to assess glucose metabolism of xenograft tumors. Following incubation with fluorescent 2-NBDG, circulating breast cancer cells in the blood samples collected from the tumor-bearing mice were collected by magnetic separation, followed by fluorescence imaging for 2-NBDG uptake by circulating breast cancer cells, and correlation of the number of hypermetabolic circulating breast cancer cells with tumor size at the time when the blood samples were collected.

Results

Human breast cancer xenograft tumors derived from MDA-MB-231, BT474, or SKBR-3 cells were visualized on near-infrared fluorescence imaging of the tumor-bearing mice injected with 2-DG 750. Hypermetabolic circulating breast cancer cells with increased uptake of fluorescent 2-NBDG were detected in the blood samples from tumor-bearing mice and visualized by fluorescence imaging, but not in the blood samples from normal control mice. The number of hypermetabolic circulating breast cancer cells increased along with growth of xenograft tumors, with the number of hypermetabolic circulating breast cancer cells detected in the mice bearing MDA-MB231 xenografts larger than those in the mice bearing BT474 or SKBR-3 xenograft tumors.

Conclusions

Circulating breast cancer cells with increased uptake of fluorescent 2-NBDG were detected in mice bearing human breast cancer xenograft tumors by fluorescence imaging, suggesting clinical use of 2-NBDG as a tracer for fluorescence imaging of hypermetabolic circulating breast cancer cells.     

Pyrene Fluorescence as a Probe for the Monitoring of Polymerization Processes: Simultaneous DSC and Fluorescence Study

The polymerization of cyclohexylmethacrylate was monitored, over a wide temperature range, by simultaneous differential scanning calorimetry (DSC) and fluorimetry (FL); the equipment for both being optically coupled. Pyrene was used as a fluorescent probe. There was excellent agreement between the exothermic peak time obtained by DSC and the time at which the maximum gradient was obtained in the fluorescence intensity–time curves, but only for temperatures above 60°C, Activation energies for the gel effect onset were obtained for both, the conversion–time and intensity–time curves being concordant except for data at low temperatures, below 60°C. It was concluded that vitrification occurs at temperatures below 60°C and its presence was demonstrated by fluorimetry, by means of the conversion change associated with the fluorescence intensity jump_(off-on).     

Probing DNA Hybridization in Homogeneous Solution and at Interfaces via Measurement of the Intrinsic Fluorescence Decay Time of a Single Label

The hybridization of DNA oligomers including molecular beacons can be detected by measurement of either the decay time or the intensity of a single fluorescent label attached to the end of the respective oligonucleotide. The method works both in solution and solid phase and can distinguish between fully complementary and mismatch sequences as demonstrated for a 15-mer oligonucleotide and a 25-mer molecular beacon. The fluorescence lifetime method is advantageous in (a) requiring a single label (and therefore a single labeling step) only; and (b), being based on measurement of a self-referenced magnitude that is hardly affected by parameters such as fluctuations in light intensity that make measurement of intensity more prone to interferences.     

Probing the structure of nuclei with the aid of bound a hyperons

It is shown that the binding energy b of heavy nuclei can serve to derive information on the nuclear radius only in the form of a certain dependence of the nuclear radius upon the proposed width of the edge region of the nucleus. The experimental b values and the experimental values of the width of the edge region result in estimates of the parameter nuclear radius₀= 1.28 _–0.30 ^+0.42 F.Translated from Izvestiya Vysshikh Uchebnykh Zavedenii, Fizika, No. 3, pp. 82–86, March, 1976.     

Quantum Dissonance as an Indicator of Quantum Phase Transition in the XXZ Chain

We investigate the behavior of quantum dissonance in the anti-ferromagnetic XXZ spin S = 1/2 chain, which exhibits a quantum phase transition. Based on a unified view of quantum and classical correlations, quantum dissonance is analytically calculated and is compared with entanglement, discord, and classical correlations for the ground state of the system. It is found that the nearest-neighbor quantum dissonance achieves an extremum and exhibits the sharpest change at the critical point. Therefore, quantum dissonance may serve as a more efficient indicator of quantum phase transitions in the XXZ spin chain.     

5-Br-PADAP分光光度法测定手掌面镀锌工具遗留印迹的锌含量

研究2-(5-溴-2吡啶偶氮)-5-二乙氨基苯酚(5-Br-PADAP)分光光度法测定手掌面镀锌工具遗留印迹的锌含量。考察了缓冲液用量、pH、显色剂用量等反应条件对测定的影响;在优化好的条件下,建立了标准曲线并测定了不同接触时间、时间间隔条件下手掌面镀锌工具遗留印迹的锌含量。研究结果表明,pH 8.0的硼砂-硼酸缓冲液4 mL,1 g·L-1的5-Br-PADAP溶液0.2 mL以及10%曲拉通-X-100溶液1 mL为最佳测定条件,在此条件下,Zn2+在0~14 μg范围内呈良好的线性关系,回归方程为y=1.851 34x+0.002 29;在接触时间10 s~5 min内,手掌面镀锌工具遗留印迹锌含量呈上升趋势,在5~10 min内趋于平缓,在接触时间5 min时已基本达到饱和状态,锌含量在0.425~2.377 μg·cm-2;手掌面镀锌工具遗留印迹锌含量在间隔时间0~2 h急剧下降,在2~7 h内下降的幅度逐渐变小,当间隔时间达到7 h时,锌含量仅为0.188 μg·cm-2,与间隔0 h锌含量相比减少了约90%。因此,微量金属显现实验一定要尽早进行。此外,不同间隔时间条件下手掌面锌含量的变化规律与喷洒显现实验中遗留印迹强度变化并非完全对应,这说明手掌面上的锌含量并非唯一可以影响不同间隔时间条件下遗留印迹强度的因素。首次在实验中证实了Zn2+与蛋白质结合后可以被络合能力更强的5-Br-PADAP分子夺取出来而显色的假设。运用此种宏观微观相结合的方法进行研究,有利于对微量金属显现实验中各影响因素机理的探讨,为进一步的研究奠定了基础。     

Fluorescence Studies of the Acetylcholine Receptor: Structure and Dynamics in Membranes and Cells

The nicotinic acetylcholine receptor (AChR) is the archetype member of the superfamily of ligand-gated ion channels that mediate fast intercellular communication in response to endogenous neurotransmitters. Here I review a series of biophysical studies on the AChR protein, with particular focus on the interactions of the macromolecule with its lipid microenvironment. Fluorescence recovery after photobleaching and phosphorescence anisotropy studies of the membrane-embedded AChR have contributed to our understanding of the translational and rotational dynamics of this protein in synthetic lipid bilayers and in the native membrane. Electron spin resonance studies led to the discovery of a lipid fraction in direct contact with the AChR with rotational dynamics 50-fold slower than that of the bulk lipids. This lipid belt region around the AChR molecule has since been intensively studied with the aim to define its possible role in the modulation of receptor function. The polarity and molecular dynamics of solvent dipoles—mainly water—in the vicinity of the lipids in the AChR membrane have been studied exploiting the amphiphilic fluorescent probe Laurdan's exquisite sensitivity to the phase state of the membrane, and Förster-type resonance energy transfer (FRET) was introduced to characterize the receptor-associated lipid microenvironment. FRET was used to discriminate between the bulk lipid and the lipid belt region in the vicinity of the protein. Further refinement of this topographical information was provided by the parallax method using phospholipid spin labels. The AChR-vicinal lipid is in a liquid-ordered phase and exhibits a higher degree of order than the bulk bilayer lipid. Changes in FRET efficiency induced by fatty acids, phospholipid, and cholesterol also led to the identification of discrete sites for these lipids on the AChR protein. I also illustrate the extension of Laurdan fluorescence studies to intact living cells heterologously expressing AChR in a brief section devoted to recent studies using two-photon fluorescence microscopy. The spatial resolution afforded by the two-photon optical sectioning of the cell in combination with the advantageous spectroscopic properties of Laurdan are exploited to obtain information on the physical state of the lipid environment of the membrane. Finally, the application of site-specific labeling and steady-state fluorescence spectroscopy to probe the location of AChR membrane-embedded domains is illustrated. The topography of the pyrene-labeled Cys residues in transmembrane domains M1, M4, M1, and M4 with respect to the membrane was determined by differential fluorescence quenching with lipid-resident spin-labeled probes. Cys residues were found to lie in a shallow position. For M4 segments, this is compatible with a linear -helical structure, but not so for M1, for which classical models locate Cys residues at the center of the hydrophobic stretch. The transmembrane topography of M1 can be rationalized on the basis of the presence of a substantial amount of nonhelical structure and/or of kinks attributable to the occurrence of the evolutionarily conserved proline residues. The latter is a striking feature of M1 in the AChR and all members of the rapid ligand-gated ion channel superfamily.     

EB荧光探针法研究多粘菌素B与DNA的作用方式

本文以溴化乙锭（EB）为荧光探针，对多粘菌素B（Polymyxin B sulfate)与DNA的作用机制进行了研究。对荧光光谱法、紫外光谱法、荧光偏振法、热变性等方面的研究结果表明：在pH=7.4的溶液中PB与DNA双螺旋碱基对之间存在嵌插作用，在一定离子强度下该嵌插作用会减弱。此外，还存在着非特性的静电作用。