水中微囊藻毒素高效液相色谱检测与前处理条件优化

通过对前处理过程中关键环节进行单因素不同水平比较,以及运用正交试验设计进行多因素综合分析,优选出洗脱剂、淋洗剂、固相萃取柱、浓缩定容方式等因素的最优试验方案,建立实用简便易行的一般水样前处理流程.微囊藻毒素高效液相色谱进样分析时,选用甲醇-0.05%三氟乙酸水溶液作为流动相进行梯度洗脱,对于MC-RR和MC-LR取得较好分离效果,水样检出限可达0.02 μg/L.优化后的前处理、检测方法,对MC-RR,MC-LR平均回收率分别为88.9%和92.1%.另外,对于总(胞内胞外)微囊藻毒素的检测前处理,冰乙酸处理法优于其他处理法.优化后的方法已成功应用于不同水域环境样本的藻毒素分析.     

Induction of vanadium accumulation and nuclear sequestration causing cell suicide in human Chang liver cells

Very little is known about the modulation of vanadium accumulation in cells, although this ultratrace element has long been seen as an essential nutrient in lower life forms, but not necessarily in humans where factors modulating cellular uptake of vanadium seem unclear. Using nuclear microscopy, which is capable of the direct evaluation of free and bound (total) elemental concentrations of single cells we show here that an NH₄Cl acidification prepulse causes distinctive accumulation of vanadium (free and bound) in human Chang liver cells, concentrating particularly in the nucleus. Vanadium loaded with acidification but leaked away with realkalinization, suggests proton-dependent loading. Vanadyl(4), the oxidative state of intracellular vanadium ions, is known to be a potent source of hydroxyl free radicals (OH^.). The high oxidative state of nuclei after induction of vanadyl(4) loading was shown by the redox indicator methylene blue, suggesting direct oxidative damage to nuclear DNA. Flow cytometric evaluation of cell cycle phase-specific DNA composition showed degradation of both 2N and 4N DNA phases in G₁, S and G₂/M cell cycle profiles to a solitary 1N DNA peak, in a dose-dependent manner, effective from micromolar vanadyl(4) levels. This trend was reproduced with microccocal nuclease digestion in a time response, supporting the notion of DNA fragmentation effects. Several other approaches confirmed fragmentation occurring in virtually all cells after 4 mM V(4) loading. Ultrastructural profiles showed various stages of autophagic autodigestion and well defined plasma membrane outlines, consistent with programmed cell death but not with necrotic cell death. Direct intranuclear oxidative damage seemed associated with the induction of mass suicide in these human Chang liver cells following vanadium loading and nuclear sequestration.     

平衡冷冻后红细胞胞内未冻水的状态

基于Boyle—Van’t Hoff关系设计了2种独立的实验方案，研究了红细胞内束缚水的状态．第一种方案使用电子粒子计数仪(EPC)测定了室温下在不同浓度的NaCl溶液中取得渗透平衡的红细胞的体积(V)，其中包括红细胞的等渗体积V0(与0．9％的NaCl溶液取得渗透平衡的红细胞体积)．第二种方案使用差示扫描量热仪(DSC)测定了不同压积比的红细胞悬液慢速冷冻的放热量．通过理论模型计算，进一步得到红细胞的最终平衡体积(Hf)．平衡冷冻的最终体积与细胞内非渗透性体积(Vb)存在差别，这一差别被认为是细胞内束缚水的体积．实验结果表明：(1)两种实验方法等效；(2)平衡冷冻后胞内几乎没有自由水，亦即红细胞内束缚水只有一种状态，为结合水．     

盐胁迫对鲁氏酵母菌生理特性的影响

研究了盐胁迫对鲁氏酵母菌(Zygosaccharomyces rouxii CGMCC 3791)细胞生理特性的影响,结果表明,盐胁迫使细胞生长受到抑制,单位(每克)菌体乙醇含量从11.64mg增加到19.20mg(120g/L NaCl)。分析细胞胞内pH值和胞内活性氧(ROS)水平,结果表明:盐胁迫使细胞胞内pH值水平降低,胞内活性氧(ROS)水平增加,同时胞内抗氧化酶系(过氧化氢酶、超氧化物歧化酶、过氧化物酶,以及谷胱甘肽过氧化物酶)的活力提高,从而抵御由盐胁迫引起的氧胁迫。研究了盐胁迫对鲁氏酵母菌胞内谷胱甘肽(GSH)含量的影响,结果表明,随着盐质量浓度增加,胞内GSH含量增加,在120g/L盐质量浓度下,GSH含量显著增加了73.4%。考察了外源添加GSH对细胞生长的影响,发现添加0.5g/L GSH的鲁氏酵母菌,在120g/L盐质量浓度下生物量提高了15%。本研究可对深入认识鲁氏酵母菌耐盐生理机制及进一步提高其耐盐性能提供理论支持。     

Calculation of the intracellular elastic modulus based on an atomic force microscope micro-cutting system

Better understanding of variations in the mechanical properties of cancer cells could help to provide novel solutions for the diagnosis,prevention,and treatment of cancers.We therefore developed a calculation model of the intracellular elastic modulus based on the contact pressure between the silicon tip of an atomic force microscope and the target cells,and cutting depth.Ovarian cells(UACC-1598) and colon cancer cells(NCI-H716) were cut into sequential layers using an atomic force microscope silicon tip.The cutting area on the cells was 8μm×8μm,and the loading force acting on the cells was increased from 17.523 to 32.126μN.The elastic modulus distribution was measured after each cutting process.There were significant differences in contact pressure and cutting depth between different cells under the same loading force,which could be attributed to differences in their intrinsic structures and mechanical properties.The differences between the average elastic modulus and surface elastic modulus for UACC-1598 and NCI-H716 cells were 0.288±0.08 kPa and 0.376±0.16 kPa,respectively.These results demonstrate that this micro-cutting method can be used to measure intracellular mechanical properties,which could in turn provide a more accurate experimental basis for the development of novel methods for the diagnosis and treatment of various diseases.     

乙酰水杨酸对大鼠DRG神经元膜电位的影响

应用细胞内微电极记录技术，观察乙酰水杨酸对大鼠背根神经节神经元膜电位的影响，结果发现：大多数细胞(89．33％)在滴加乙酰水杨酸(10^-9～10^-3mol／L)后可引起浓度依赖性的超极化反应，并对其可能镇痛机制进行了讨论。     

Regulatory aspects of low intensity photon emission

Summary Photon emission from unicellular and multicellular organisms has been a subject of study for many decennia. In contrast to the well-known phenomenon of bioluminescence originating in luciferin-luciferase reactions, low intensity emission in the visible region of the electromagnetic spectrum has been found in almost every species studied so far. At present, the nomenclature of this phenomenon has not crystallized and it is referred to by a variety of names, such as mitogenetic radiation²⁹, dark luminescence⁷, low-level chemiluminescence^{20, 36}, and biophotons⁵⁷. Particular attention has been focussed on the relationship between photon emission and the regulation of various aspects of cellular metabolism, although in many cases quantitative data are still lacking. Throughout the history of this field of research the question of a functional biological role of the low intensity emission has been repeatedly raised; this is reflected, for instance, in the heterogeneity of the terms used to describe it. The discussion concerns the possible participation of photons of low intensity in intra- and intercellular communication. This paper reviews literature on the metabolic regulation of low intensity emission, as well as the regulation of photon emission initiated by external light. Furthermore, recent data are discussed with respect to a possible biocommunicative function of low intensity photon emission.     

酵母胞内核黄素含量测定方法的研究

对测定核黄素常用的3种方法,即高效液相色谱法、荧光光度法和分光光度法进行比较.结果表明,对于标准溶液3种方法均具有良好的线性关系和重现性,但在酵母提取液中,高效液相色谱法和分光光度法的回收率较差,测定准确率低.荧光光度法测定准确性高,稳定性强,为测定酵母胞内核黄素的最佳方法.     

~(31)P核磁共振法研究完整脏器细胞内的pH值

本工作对老鼠的心、肝、肾等不同脏器进行了(81)~P核磁共振的研究,并采用六甲基磷酰三胺为外标,测定了各种正常脏器细胞内的pH数值,同时也观察了在冷缺血、热缺血的不同条件下,离体脏器细胞内pH和无机磷酸盐浓度的变化。