微弱发光分析技术应用实例(六)——发光分析与肿瘤研究

微弱发光分析技术已经用于肿瘤学研究,骨肿瘤病人和正常人的血液和尿液的发光强度使用BPCL型微弱发光测量仪进行了测量.结果指出,骨肿瘤病人血液和尿液的发光强度高于正常人（P＜0.05）.骨肿瘤病人尿液的发光强度在手术之后明显降低（P＜0.05）.裸鼠血液和各种脏器的微弱发光测量结果表明,荷瘤之后,各个脏器的发光强度显著增加.     

Latent Osteomalacia in Epileptic Patients on Anticonvulsants

The bone mineral content was measured in 10 epileptic patients on long-term treatment with phenytoin before and during treatment with vitamin D. None of the patients showed biochemical signs of osteomalacia. Initially subnormal values for bone mineral content were found, which increased significantly during treatment. The results suggest the occurrence of latent osteomalacia in a fairly high proportion of epileptic patients on anticonvulsants.     

盐胁迫下豌豆幼苗的超弱发光

通过钠盐对豌豆 (pisumsativum)萌发时超弱发光影响的比较研究表明 ,在等渗条件下豌豆幼苗受激发光强度顺序为Na2 CO3 >NaCl>Na2 SO4 ;Na2 CO3 盐胁迫对植物的伤害程度高于其他盐类 ;多盐比单盐存在时的毒害小 ,受激发光作用降低。利用超弱发光方法 ,可以分析盐害对植物的作用 ,进一步搞清胁迫条件下植物生理反应的变化规律。     

Active oxygen generation in γ-irradiated whole seedlings of Cicer arietinum L.

Whole seedlings of Cicer arietinum L. when exposed to 819 Gy γ-radiation, were found to emit ultraweak intensity luminescence. The phenomenon was oxygen-dependent, and the intensity of emission could be suppressed by post-irradiation treatment with catalase, superoxide dismutase, and ascorbic acid. Deuterium oxide and luminol amplified the emission intensity. These results suggest that singlet oxygen is a cause of the emission during the post-irradiation phase.     

Effect of Vitamin D on Bone Mineral Mass in Normal Subjects and in Epileptic Patients on Anticonvulsants: A Controlled Therapeutic Trial

The bone mineral mass was estimated by photon absorptiometry in 23 epileptic patients on long-term treatment with phenytoin and in 20 normal subjects before and during treatment with vitamin D or placebo.Initially, subnormal values of bone mineral mass were found in the epileptic patients. The group of epileptic patients treated with vitamin D showed a significant increase in bone mineral mass. The group of epileptic patients treated with placebo and the normal subjects treated with vitamin D or placebo showed no change in bone mineral mass.     

Optically stimulated luminescence dating of cave deposits at the Xiaogushan prehistoric site, northeastern China

The Xiaogushan cave site is one of the most important prehistoric sites in North China. The stone and bone artifacts found in the cave are similar to European contemporaneous artifacts. Cave deposits consist of five layers that have been dated from 46,353 ± 1179 to 4229 ± 135 cal. yr BP, using radiocarbon dating techniques on charcoal and bone samples collected from Layers 2-5. In this paper, optically stimulated luminescence (OSL) techniques were applied to date six samples taken from Layers 1-3. The luminescence properties of the fine-grained and coarse-grained quartz extracts indicate that the materials are suitable for OSL dating using a single-aliquot regeneration-dose (SAR) protocol. The OSL ages obtained are broadly consistent with the stratigraphy and the associated calibrated radiocarbon ages. The dating results show that the cave was first occupied by humans about 70 ka. The human occupation of the cave may be related to climate change. An occupation hiatus is inferred to between ∼17 to ∼10 ka. The stone and bone artifacts found in Layers 2 and 3 may indicate the Middle to Upper Paleolithic transitions in the region.     

Characteristics of the intrinsic luminescence of Actinomyces olivocinereus--producer of heliomycin]

Some characteristics of UV-induced luminescence were studied with Actinomyces olivocinereus producing the antibiotic heliomycin. The luminescence of the growth medium was found to be caused not by heliomycin, but by some other factors. The luminescence of heliomycin in the colonies was quenched as a result of its screening with melanin pigments located in a layer between the aerial and substrate mycelium.     

Relation between slow delayed light emission and acid-base triggered luminescence in chloroplasts

Acid-base triggered luminescence in relation to slow delayed light emission (> 3 s) was studied in chloroplasts. After analyzing their time courses, the acid-base induced luminescence curve was found to return to the original curve of delayed light emission. Peaks of the acid-base triggered luminescence induced after various darkness periods following preillumination decreased parallel to the time course of delayed light emission without base treatment. 3-(3,4-Dichlorophenyl)-1,1-dimethylurea enhanced both the delayed light emission and acid-base induced luminescence, while carbonyl cyanide m-chlorophenylhydrazone inhibited both. Several photophosphorylation uncouplers inhibited the acid-base induced luminescence without any substantial effect on the delayed light emission. It is concluded that the acid-base triggered luminescence is not caused by the reversion of electrons from remote intermediates on the reducing side of Photosystem II. The possibility of the presence of an activation pathway for the acid-base triggered luminescence which differs from that of the delayed light emission is also discussed.     

Granulopoietic studies in acute lymphocytic leukemia of children.

Studies have been carried out on the levels of serum and urine colony stimulating activity (CSA) and peripheral blood and bone marrow colony forming cell numbers in children with acute lymphocytic leukemia (ALL) during various phases of their disease. These studies have suggested that serum and urine levels of colony stimulating factor are reduced during the inital or relapse phase of the disease compared to levels found during remission. It has also been found that the number of bone marrow colony forming cells is reduced in relapse or before treatment and elevated during remission while the number of peripheral blood colony forming cells is increased during relapse or before treatment and normal during remission. It has also been shown that mixing of serum or leukemic cells with normal human bone marrow cells inhibits colony formation.     

Measuring dose from radiotherapy treatments in the vicinity of a cardiac pacemaker

This study investigated the dose absorbed by tissues surrounding artificial cardiac pacemakers during external beam radiotherapy procedures. The usefulness of out-of-field reference data, treatment planning systems, and skin dose measurements to estimate the dose in the vicinity of a pacemaker was also examined. Measurements were performed by installing a pacemaker onto an anthropomorphic phantom, and using radiochromic film and optically stimulated luminescence dosimeters to measure the dose in the vicinity of the device during the delivery of square fields and clinical treatment plans. It was found that the dose delivered in the vicinity of the cardiac device was unevenly distributed both laterally and anteroposteriorly. As the device was moved distally from the square field, the dose dropped exponentially, in line with out-of-field reference data in the literature. Treatment planning systems were found to substantially underestimate the dose for volumetric modulated arc therapy, helical tomotherapy, and 3D conformal treatments. The skin dose was observed to be either greater or lesser than the dose received at the depth of the device, depending on the treatment site, and so care should be if skin dose measurements are to be used to estimate the dose to a pacemaker. Square field reference data may be used as an upper estimate of absorbed dose per monitor unit in the vicinity of a cardiac device for complex treatments involving multiple gantry angles.     

光动力学疗法的单态氧剂量学研究进展

单态氧(1O2)是光动力学疗法(photodynamic therapy,PDT)过程中Ⅱ型光化学反应的主要细胞毒性物质.通过直接测量1O2发光强度预测PDT疗效已成为PDT剂量学研究的前沿热点,这种方法的最大优点在于能够克服现有其它剂量学方法中的光、光敏剂、氧分子,以及组织光学特性等因素之间的复杂相互影响,将PDT的...     

31P NMR and saturation transfer studies of the effect of Pb2+ on cultured osteoblastic bone cells

The mechanism of lead toxicity at the cellular level remains unknown, although an effect of lead on intracellular Ca2+ has been described. Since bone is a major target for lead, we have investigated the effect of lead on bioenergetic rates and on the intracellular free Mg2+ concentration in cultured osteoblastic bone cells. Using 31P NMR and the saturation transfer technique we have detected a sizable (18%) transfer of saturation from gamma ATP to Pi in a perfused osteoblastic osteosarcoma bone cell line, Ros 17/2.8, and have found a large (greater than 82%) reduction in the Pi----ATP rate upon treatment with 10 microM Pb2+. The NMR-measured unidirectional rate was much greater than the net rate of ATP synthesis through glycolysis and oxidative phosphorylation. By using iodoacetate we investigated the mechanism of the saturation transfer and found that it is catalyzed by the glycolytic enzyme couple glyceraldehyde-3-phosphate dehydrogenase/phosphoglycerate kinase. The net rate of glycolysis as measured by lactate production and that of oxidative phosphorylation as measured by O2 consumption were found to be significantly decreased by 18 and 74%, respectively, with lead treatment. In addition, from the chemical shifts of intracellular ATP resonances, we found a significant reduction of 21% in the intracellular free Mg2+ concentration upon Pb2+ treatment. The observed lead-induced reduction in ATP synthesis/utilization and the decrease in intracellular free Mg2+ may contribute to the impairment of bone formation during lead intoxication.     

Electrophotoluminescence and the electrical properties of the photosynthetic membrane. II. Electric field-induced electrical breakdown of the photosynthetic membrane and its recovery

Preilluminated suspensions of swollen thylakoid vesicles (‘blebs’) were exposed to uni- and bipolar pairs of identical electric field pulses of variable duration, intensity and spacing. The resulting field-stimulated luminescence (electrophotoluminescence) was used as an intrinsic, voltage-sensitive optical probe to monitor electrical phenomena at the membrane level. The application of a pair of voltage pulses of opposite polarity made it possible to produce electric changes in the membrane by the first pulse and to analyse these effects by a second pulse of opposite polarity. It was found that the relative amplitudes of the two electrophoto-luminescence signals depended on the intensity of the applied electric field and on the time interval (t*) between the two pulses. When t* varied from 0.4 to 12 ms, the second stimulated luminescence signal was at first much smaller than the first one and then increased exponentially until the two signals were equal for t* ≥ 3 ms. We analysed these differences between the two field-stimulated luminescence signals as a measure of the electrical breakdown of the membrane, induced during the first pulse. In this way a distinction between irreversible and reversible breakdown could be made with an estimation of the recovery kinetics of the reversible breakdown, which was found to be complete within 3 ms. Irreversible breakdown of the membrane was found to increase with lengthening the exposure time from 0.1 to 1.3 ms especially when applying high electric field of at least 2000 V/cm.     

牙周骨再生过程中BMP-1表达变化的临床意义

目的:观察在牙周骨再生过程中骨形态发生蛋白-1(BMP-1)的表达的变化情况及临床意义。方法:以我院收治的牙周骨缺损患者196例作为研究对象,采用常规的基础牙周治疗(龈上洁治彻底,龈下刮治,整平根面)处理后,通过引导组织再生术(GTR)技术对骨缺损进行修复的同时,植入Bio-oss人工骨材料和Bio-gide胶原膜。在治疗前和治疗后,分别测定患者血清BMP-1水平,分析BMP-1表达水平与牙周骨缺损修复的关系。结果:与治疗前相比,患者治疗后的牙周骨缺损均有所修复,所有患者在术后所有时间点的PPD和骨密度值与治疗前相比均显著改善(P0.05)。治疗6个月后,牙周骨密度出现小幅度下降,但显著高于治疗前,差异具有统计学意义(P0.05)。与治疗前相比,所有患者治疗后1 d、2 d、5 d、10 d、1个月、3个月、6个月血清BMP-1的水平均显著升高,差异具有统计学意义(P0.05),且治疗后BMP-1水平的变化与PPD呈显著负相关(P0.05),但与骨密度的相关性不显著(P0.05)。结论:血清BMP-1在牙周骨修复过程存在动态变化与患者牙周骨术后恢复相关。     

Estradiol-17β and nutritional status affect calcium balance, scale and bone resorption, and bone formation in rainbow trout, Oncorhynchus mykiss

The effects of estradiol-17β (E₂) on bone resorption and formation as well as its effects on scale resorption were investigated in rainbow trout in order to elucidate the role of the hormone in calcium mobilization from calcified tissues, and to clarify the importance of scale and bone as calcium reserves during sexual maturation. Furthermore, the effects of nutritional status on calcified tissues and E₂-induced calcium mobilization were studied. In fed as well as fasted rainbow trout, E₂ treatment increased scale osteoclastic activity measured as tartrate-resistant acid phosphatase activity, and reduced scale calcium content, suggesting that E₂ increases scale resorption in both the fed and fasted fish. Using histomorphometry, E₂ treatment was found to decrease pharyngeal bone resorption in fed, but not in fasted rainbow trout. The E₂ effect on rainbow trout bone is consistent with its physiological role in mammals and birds where E₂ has been reported to decrease bone resorption. It appears therefore that rainbow trout protect their skeleton and instead use scales as a source of calcium during E₂-induced calcium mobilization. The formation of pharyngeal bone was decreased by fasting, and the importance of the nutritional status for the activity of the bone cells in rainbow trout is therefore emphasized. Accepted: 8 May 1997     

Studies of calcium influx into squid giant axons with aequorin

Calcium-influx associated with the action potential was studied in squid giant axons using an EDTA-free preparation of aequorin as a probe. Associated with an individual action potential there was a transient increase in aequorin luminescence and the time course of this increase was examined on a kinetic basis. The luminescent response associated with a train of action potentials was compared with that expected on the basis of superposition of individual responses. The analyses of luminescence curves produced by long trains of action potentials were complicated by failure of superposition. A long pulse of inward current was found to produce a very large enhancement of luminescence.     

Effect of Na+ and K+ ions on the luminescence of intact Vibrio harveyi cells at different pH values

The bioluminescent activity of intact Vibrio harveyi cells loaded with different concentrations of NaCl and KCl at different pH values was studied. In the pH range of 6.5-8.5, the effect of Na+ was significantly higher than that of K+ at all concentrations studied. Maximum luminescent activity was observed in cells loaded with 0.68 M NaCl. When Na+ was nonuniformly distributed on the plasma membrane, the cell luminescence kinetics was nonstationary in the 20-min range: during incubation, the luminescence intensity increased at pH 6.5 and decreased at pH 8.5. The activation and damping rate constants depended on the Na+ gradient value. The maximum of luminescent activity shifted during incubation from pH 8.5 to 6.5-7.0. The luminescence kinetics in the systems with KCl was stationary; the maximum level of luminescence was observed in the pH range of 7.0-7.5. Under Na(+)-controlled conditions, the cell respiration and luminescence changed in synchronism. The protonophore CCP at a concentration of 20 microM completely inhibited luminescence at pH 6.5 and was ineffective at pH 8.5.     

Coral Luminescence Identifies the Pacific Decadal Oscillation as a Primary Driver of River Runoff Variability Impacting the Southern Great Barrier Reef

The Pacific Decadal Oscillation (PDO) is a large-scale climatic phenomenon modulating ocean-atmosphere variability on decadal time scales. While precipitation and river flow variability in the Great Barrier Reef (GBR) catchments are sensitive to PDO phases, the extent to which the PDO influences coral reefs is poorly understood. Here, six Porites coral cores were used to produce a composite record of coral luminescence variability (runoff proxy) and identify drivers of terrestrial influence on the Keppel reefs, southern GBR. We found that coral skeletal luminescence effectively captured seasonal, inter-annual and decadal variability of river discharge and rainfall from the Fitzroy River catchment. Most importantly, although the influence of El Niño-Southern Oscillation (ENSO) events was evident in the luminescence records, the variability in the coral luminescence composite record was significantly explained by the PDO. Negative luminescence anomalies (reduced runoff) were associated with El Niño years during positive PDO phases while positive luminescence anomalies (increased runoff) coincided with strong/moderate La Niña years during negative PDO phases. This study provides clear evidence that not only ENSO but also the PDO have significantly affected runoff regimes at the Keppel reefs for at least a century, and suggests that upcoming hydrological disturbances and ecological responses in the southern GBR region will be mediated by the future evolution of these sources of climate variability.     

High optical-throughput spectroscopic singlet oxygen and photosensitizer luminescence dosimeter for monitoring of photodynamic therapy

We report a high light-throughput spectroscopic dosimeter system that is able to noninvasively measure luminescence signals of singlet oxygen (¹O₂) produced during photodynamic therapy (PDT) using a CW (continuous wave) light source. The system is based on a compact, fiber-coupled, high collection efficiency spectrometer (>50% transmittance) designed to maximize optical throughput but with sufficient spectral resolution (~7 nm). This is adequate to detect ¹O₂ phosphorescence in the presence of strong luminescence background in vivo. This system provides simultaneous acquisition of multiple spectral data points, allowing for more accurate determination of luminescence baseline via spectral fitting and thus the extraction of ¹O₂ phosphorescence signal based solely on spectroscopic decomposition, without the need for time-gating. Simultaneous collection of photons at different wavelengths improves the quantum efficiency of the system when compared to sequential spectral measurements such as filter-wheel or tunable-filter based systems. A prototype system was tested during in vivo PDT tumor regression experiments using benzoporphyrin derivative (BPD) photosensitizer. It was found that the treatment efficacy (tumor growth inhibition rate) correlated more strongly with ¹O₂ phosphorescence than with PS fluorescence. These results indicate that this high photon-collection efficiency spectrometer instrument may offer a viable option for real-time ¹O₂ dosimetry during PDT treatment using CW light.     

Chemiluminescence in a stimulated polymorphonuclear leukocytes--luminol system: suppression by thiols

The effect of some scavengers of thiol nature, which eliminate all reactive oxygen species and oxidants with reactive chlorine, on the luminol-enhanced chemiluminescence of polymorphonuclear leukocytes was studied. The use of two scavengers of this type (penetrating and not penetrating into the cell) made it possible to separate the luminescence of cell structures from the luminescence generated by oxidants in the surrounding medium. It was found that about a half of luminol luminescence is due to its oxidation in the medium surrounding the cell, and it is completely inhibited by the nonpenetrating reduced glutathione. The cell itself is a source of a considerable portion of luminescence, and this luminescence is quenched by penetrating sulfhydryl compounds such as dithiothreitol and N-acethyl cysteine. Reduced glutathione, which penetrates into cells and whose action is due only to the sulfhydryl group, is recommended as a candidate for the selective neutralization of extracellular oxidants.