Molecular characterization of a K+ channel blocker in the scorpionButhus martensii Karsch

K⁺ channel blockers of scorpion venoms are of important value in studying pharmacology and physiology of specific K⁺ channel of cells. Based on the amino acid sequences of BmP01 previously characterized as a small-conductance Ca²⁺-activated K⁺ channel blocker, two “back to back” degenarate primers have been designed and synthesized for inverse PCR strategy, its full-length cDNA has been cloned from the venom gland of the Chinese scorpionButhus martensii. The cDNA is composed of 3 parts: 5′ UTR, ORF and 3′ UTR. The flanking sequence of translation initiation codon ATG is AAAATGA, which is highly conserved in scorpion Na⁺ channel toxin and protozoan genes, suggesting that these genes may have followed a common mechanism for translation initiation. The 3′ UTR contains poly(A) signal AATAAA. The open reading frame encodes a precursor of 57 residues with a signal peptide of 28 residues and a mature peptide of 29 residues. The signal peptide is rich in hydrophobic amino acid residues and its length is significantly different from that of the determined scorpion Na⁺ channel toxin. The deduced amino acid sequence of mature peptide is completely consistent with BmP01 previously determined by primary structure analysis.     

Na+-permeable channels of human sperm membrane reassembled into giant liposome

Previous data showed that a Na⁺-transmembrane flux was accompanied with acrosome reaction of sperm. However, the electrophysiological recording and characterization of Na⁺ current in human sperm membrane have not been yet reported. In the present investigation, membrane proteins extracted from human sperms were reassembled into liposome bilayer, and then the liposomes were fused by dehydration-rehydration into giant liposomes with the diameter of more than 10 ώm. By patch clamping the giant liposomes two kinds of single channel currents were recorded in a NaCl solution system. Both of them were Na⁺-carried, TTX-sensitive and strongly rectifying, but with different unit conductance and open probability. Moreover, bursting activity and channel-substates as well as two open time constants were observed in the larger channel.     

HERG1 K+ channels on the leukemic cells mediated angiogenesis in vitro

Human ether-a-go-go-related gene （HERG1） K^＋ channels are overexpressed in leukemia, which contributes to neoangiogene- sis. The purpose of this study was to investigate the role of HERG1 K^＋ channels on leukemia angiogenesis. We cultured human umbili- cal vein endothelial cells （HUVECs） in conditioned media, which were derived from leukemic cells with or without E-4031, a HERG1 K^＋ channel special inhibitor. The HUVECs proliferation was mea- sured using CCK-8 assay and migration by a Trans-well. Endothelial tube formation was investigated using Matrigel. Vascular endothelial growth factor （VEGF） levels were tested by ELISA and VEGF mRNA expression using RT-PCR. Our results revealed that blocking HERG1 K^＋ channels could inhibit leukemia-induced HUVECs pro- liferation, migration, and tube formation in vitro. The results sug- gested that HERG1 K~ channels could increase leukemia angio- genesis. Furthermore, blockage of HERG1 K^＋ channels could also decrease leukemic cells secreting VEGF and expressing VEGF mRNA. HERG1 K^＋ channels have a promoting effect on leukemia angiogenesis, and the possible mechanism may be that HERG1 K^＋ channels enhance VEGF expression. Thus, HERG1 K4 channel is a potential target of antiangiogenesis in leukemia.     

模拟微重力大鼠T淋巴细胞发育异常的初步研究

对模拟空间微重力条件下大鼠T细胞发育受阻的神经免疫调节机理进行初步探究.采用流式细胞检测技术测定模拟微重力大鼠胸腺中不同发育阶段T淋巴细胞的数量变化,用ELISA法测定胸腺组织中去甲肾上腺素(NE)和内源性糖皮质激素(GC)的含量.结果表明模拟微重力模型大鼠出现明显胸腺萎缩(p<0.05),CD3⁺CD4⁺CD8^-和CD3⁺CD4^-CD8⁺T细胞数量明显减少(p<0.05);模拟微重力大鼠胸腺中NE和GC含量有升高的趋势.模拟微重力大鼠胸腺中发生了从双阳性T细胞向单阳性T细胞的发育阻滞,这种异常可能与神经内分泌免疫调控相关.     

Mechanism for chronic pain generation

Neuropathic pain and the other abnormalities of sensation induced by axon injury or by peripheral nerve inflammation should result from functional compensations of the injured neurons during their regeneration. Ectopic distribution of proteins related to Na⁺, K⁺ and Ca²⁺ channels as well as of receptors on both membranes of injured axon and its cell body becomes a main pacemaker from which spontaneous ectopic afferent of primary sensatory neurons and crosstalk between neurons occur. Abnormal ectopic afferent activities lead to disorders of the sensation, such as hyperalgesia, allodynia, spontaneous pain and paraesthesia. Administration of some ion channel agents and/or α2-adrenergic blockers has shown efficiency in preventing neuropathic pain development and in relieving neuropathic pain.     

固液冲蚀作用下的四通流道设计及分析

在油气钻采过程中,为了提高压裂效率,部分井场会采用一种高压四通管汇连接装置。基于液固两相流和冲蚀磨损理论,建立了流道夹角分别为0°～14°在不同工况下的三维模型,利用fluent软件探究水力压裂下高压四通管的冲蚀磨损特性。结果表明：传统的直流道四通不论在何种工况下冲蚀集中区域都在四通交汇的相贯线上,随着角度增加,冲蚀集中区域由相贯线及附近壁面变为出口端圆柱面;通过对流体不同工况下的分析表明,不同的流道夹角下,流体冲蚀影响各不相同。单出口工况下,较小流道夹角有效的降低了冲蚀率,随着角度继续增加,管壁的最大冲蚀率平缓增长;双出口工况下,最大冲蚀率也会有效降低,但随着夹角继续增加至5°,最大冲蚀率不会继续降低,而呈现增长的趋势;三出口工况下四通,最大冲蚀率降低效果并不明显,且流道夹角由3°增加到14°,最大冲蚀率增长幅度很大。     

飞秒激光场中甲醇产物离子的角分布

利用直流切片离子成像技术结合飞行时间质谱技术,我们研究了甲醇(CH_3OH)分子在800 nm飞秒激光场中的多光子解离和库仑爆炸过程.获得了甲醇分子在光场中的飞行时间质谱和产物离子的切片影像,分析得到了不同库仑爆炸通道碎片离子的动能分布和角度分布.本文对比分析了C-O键断裂产生H_2O~+的H转移通道和产生OH~+的非H转移通道的角分布,计算了不同激光强度下两个通道的各向异性参数α_2和(cos~2θ),并根据产物离子角度分布与激光强度和离子价态的关系,揭示了甲醇分子在飞秒光场中的准直机制为动力学准直.     

新型LED红色荧光材料Ca_xSr_(1-x-y-z)MoO_4:yEu~(3+);zNa~+的制备和表征

利用Na+为电荷补偿剂制备CaxSr1-x-y-zMoO4:yEu3+,zNa+红色LED荧光粉体.研究了电荷补偿剂的用量、基质材料中Ca2+的含量、合成温度、反应时间及发光中心Eu3+摩尔分数对荧光材料晶体结构和发光性能的影响.结果表明:最佳工艺条件为电荷补偿剂摩尔分数为6%,Eu3+摩尔分数为8%,Ca2+摩尔分数为60%,反应温度900℃和反应时间2 h.光谱测试结果表明,该荧光材料可被311,395或465 nm有效激发,发射峰在616 nm.     

Curare can open and block ionic channels associated with cholinergic receptors

Curare has long been regarded as a typical competitive antagonist of acetylcholine (ACh) at the vertebrate neuromuscular junction. Recently, however, it has been shown that curare can also block the channels opened by ACh at the frog neuromuscular junction as well as on rat and Aplysia neurones; moreover, curare is able to depolarize rat myotubes and thus behaves as an agonist for the cholinergic receptor of this preparation (see ref. 6). Using the single channel recording technique, we have now found that, on rat myotubes, curare can both open and block in the same cell the channels controlled by the cholinergic receptor.     

一种中药复方对小鼠脾脏淋巴细胞PD-1/PD-L1表达的影响

为了探讨中药复方对小鼠脾脏淋巴细胞PD-1/PD-L1的表达情况,以及CD3~+CD4~+和CD3~+CD8~+T淋巴细胞比例的影响,采用传统熬制中药的方法得到中药复方提取液,按低、中、高3个浓度对小鼠灌胃给药(1次/d),2周后,采用脑脊髓脱臼法处死小鼠,利用Real-time PCR和流式细胞仪检测脾脏PD-1/PD-L1的表达情况以及淋巴细胞亚型的变化。结果表明,随着中药提取液浓度的增加,脾脏淋巴细胞PD-1的表达显著降低,而PD-L1的表达无显著变化;CD3~+CD4~+型和CD3~+CD8~+型T淋巴细胞的比例均显著增加。所研制的中药复方能抑制淋巴细胞PD-1的表达,激活免疫辅助细胞和效应细胞,有望应用于抗肿瘤的免疫治疗。     

Nitric oxide suppresses stomatal opening by inhibiting inward-rectifying K<Stack><Subscript>in</Subscript><Superscript>+</Superscript></Stack> channels in <Emphasis Type="Italic">Arabidopsis</Emphasis> guard cells

We explore nitric oxide （NO） effect on K^＋in, channels in Arabidopsis guard cells. We observed NO inhibited K^＋in, currents when Ca^2＋ chelator EGTA （Ethylene glycol-bis（2-aminoethylether）-N,N,N′,N;tetraacetic acid） was not added in the pipette solution; K^＋in currents were not sensitive to NO when cytosolic Ca^2＋ was chelated by EGTA. NO inhibited the Arabidopsis stomatal opening, but when EGTA was added in the bath solution, inhibition effect of NO on stomatal opening vanished. Thus, it implies that NO elevates cytosolic Ca^2＋ by activating plasma membrane Ca^2＋ channels firstly, then inactivates K^＋in, chartnels, resulting in stomatal opening suppressed subsequently.     

Inverse relationship of the durations of adjacent open and shut intervals for C1 and K channels

Ion channels in cell membranes, whether voltage-dependent or activated by ligands, make repeated transitions among open and shut states during activity. Information about the number of states and the transitional pathways between them can be obtained from the durations of open and shut intervals, as transitions to states of different lifetimes result in intervals of different mean durations. If there is only one open conformation, or state, then the durations of open intervals would be independent of the durations of adjacent shut intervals. On the other hand, if a channel has two or more open states with different mean lifetimes, and if each open state is entered directly from a different shut state with a different mean lifetime, then the open intervals should be related to the adjacent shut intervals. We now report that the durations of adjacent open and shut intervals for both a C1 channel and a large conductance Ca-activated K channel in skeletal muscle are inversely related; shorter open intervals are adjacent to longer shut intervals. These findings indicate that two or more shut states make direct transitions to two or more open states, and suggest that the lifetimes of adjacent open and shut states are inversely related.     

Study the effects of metallic ions on the combination of DNA and histones with molecular combing technique

The effects of monovalent (Na^ , K^ ) and divalent (Mg^2 , Ca^2 , Mn^2 ) ions on the interaction between DNA and histone are studied using the molecular combing technique. λ-DNA molecules and DNA-histone complexes incubated with metal cations (Na^ , K^ , Mg^2 , Ca^2 , Mn^2 ) are stretched on hydrophobic surfaces, and directly observed by fluorescence microscopy. The results indicate that when these cations are added into the DNA solution, the fluorescence intensities of the stained DNA are reduced differently. The monovalent cations (Na^ , K^ ) inhibit binding of histone to DNA. The divalent cations (Mg^2 , Ca^2 , Mn^2 ) enhance significantly the binding of histone to DNA and the binding of the DNA-histone complex to the hydrophobic surface. Mn^2 also induces condensation and aggregation of the DNA- histone complex.     

Monitoring the binding of metal cations and histones to DNA in real time using fluorescence assays

The binding of cations (Na +,K +,Mg 2+,Ca 2+,Mn 2+) and histones to DNA can be studied using fluorescence assays.Here,we measured the fluorescence intensity and fluorescence anisotropy of DNA and DNA-histone complexes in the presence of cations.We demonstrate that when different cations are added into a DNA solution,the fluorescence intensities of the stained DNA are reduced by different amounts.Compared with divalent cations,monovalent cations had a weaker effect on fluorescence intensity and fluorescence anisotropy.Divalent (Mn 2+,Mg 2+,Ca 2+) cations markedly enhanced the fluorescence anisotropy of DNA.The binding modes of monovalent and divalent cations to DNA may be different.Divalent cations can change the structure of DNA molecules,or promote the assembly of DNA strands.The addition of histones causes DNA condensation,which mostly occurs during the first few seconds.Cation binding to DNA is abrupt,and is much faster than that of histones.     

Summer monsoon and dust signals recorded in the Dasuopu firn core, central Himalayas

In September 1997, a 15-m firn core was recovered from an elevation of 7 000 m a. s.l. from the Dasuopu Glacier in the central Himalayas. The analysis of δ18O values and major ion (Ca2+ , Mg2+ , NH4+ , SO42- and NO3-) concentrations shows that average annual accumulation is 0.75 m (water equivalent) in the Dasuopu firn core. The seasonal variations of δ18O values and major ion concentrations in the core indicate that present summer monsoon and dust signals are recorded with high-resolution in the Dasuopu Glacier. δ18O in precipitation are controlled by amount effect in summer monsoon season, more negative δ18O is representative of summer monsoon signal in snow layers. Higher concentrations of Ca2+ , Mg2+ , SO42- and NO3-are dominated by spring dust storm imput derived from the arid and semi-arid desert regions in central Asia. Also EOF analysis verifies that high spring concentrations of major ions are consistent. Due to the possibly different sources, the secondary variations of NH4+ and NO3- are negatively relevant with that of Ca2+ and Mg2+ .     

Photoluminescence from Si-based SiNxOy films

B⁺, C⁺, Si⁺, and As⁺ with dose of 5 × 10¹⁶ cm⁻² were implanted into SiN_xO_y, films grown on crystalline silicon by plasma-enhanced chemical vapor deposition. The ion-implanted samples exhibit their photoluminescence with different intensities and different peak wavelengths after thermal annealing. Especially, the C⁺-implanted SiN_xO_y, films show very intense photoluminescence at green-yellow color region.     

Reduced Na+ and K+ permeability of K+ channel in plasma membrane isolated from roots of salt-tolerant mutant of wheat

The Na⁺ and K⁺ permeability of K⁺ channel in plasma membrane, isolated from roots of the salt-tolerant mutant of wheat, was lower than that of wild type in 100 mmol/L KCl and NaCl solution. The opening frequency of K⁺ channel of the mutant reduced more significantly than that of wild type in two kinds of solution mentioned above. It is assumed that the reduction of opening frequency mainly contributes to the Na⁺ and K⁺ permeability of K⁺ channel of the mutant. The electric conductance of single-channel of the mutant was similar to that of wild type and the main difference between them was exhibited as the opening frequency. Their K⁺/Ka⁺ selectivity of K⁺ channel had no significant difference. The K⁺/Na⁺ selectivity of the mutant and wild type was 3.35 and 3.18 respectively.     

The effects of S4 segments on the voltage-dependence of inactivation for Cav3.1 calcium channels

T-type calcium channels exhibit fast voltage-dependent inactivation, for which the underlying struc- ture-function relationship still remains unclear. To investigate the roles of S4 segments in volt- age-dependent inactivation of T-type calcium channels, we created S4 replacement chimeras between Cav3.1 calcium channels (fast voltage-dependent inactivation) and Cav1.2 calcium channels (little voltage-dependent inactivation) by replacing S4s in Cav3.1 with the corresponding regions in Cav1.2. Wild type and chimeric channels were expressed in Xenopus oocytes and channel currents were re- corded with two-electrode voltage-clamp. We showed that replacing S4 region in domain I shifted voltage-dependence for inactivation of Cav3.1 to the left, and the V0.5 inact and kinact value were signifi- cantly changed. However replacing S4s in domains II―IV had no effects on the voltage-dependent in- activation properties. These results suggest that the roles of S4 segments in domains I―IV are different, and S4 in domain I is likely to be involved in voltage-dependent inactivation process. Its movement during membrane depolarization may trigger a conformational change in the inactivation gate.     

壳聚糖絮凝剂处理废水中的Ag+

研究了利用壳聚糖絮凝剂絮凝除Ag的方法.考察了pH值、离子溶液浓度、絮凝剂用量以及絮凝时间对去除率的影响.实验结果表明:壳聚糖絮凝除银的适宜pH为7~8;当pH=7~8时,用壳聚糖吸附银离子,水样银质量浓度低于20 mg/L时,除银率达100%.即使高浓度含银水样,其除银率也在99.9%以上;絮凝除银时间4 h为宜,其去除率达99.98%;壳聚糖的加入量过多过少都会影响除银效果.分析了壳聚糖絮凝除银的机理.     

A novel multi-functional cell-based microphysiometer

This paper presents a novel multi-functional microphysiometer for simultaneous measurements of several extracellular ion concentrations and action potential measurement in living cells based on MLAPS (multi-light addressable potentiometric sensor). In the microphysiometer, sorts of sensitive membranes are illuminated in parallel with n light sources at working frequencies, and the response amplitudes of each frequency component can be measured on-line by parallel processing algorithm. In the experiments, the relations of the extracellular environmental H⁺, Na⁺, K⁺, Ca²⁺ under the effects of western medicines (dilantin, phenobarbital sodium) and Chinese drugs (scutellaria, medlar, hemlock parsley) were analyzed, and the effects of several drugs were evaluated. Moreover, the action potential signals of different cell types (cardiac myocytes and neurons) could be measured and analyzed by LAPS. By detecting these parameters, the system can monitor the real-time process of the cell metabolism and action potential, observe the functional responses of different kinds of membrane-bound receptors, and evaluate the activities of drugs.