新型tacrine双联体对培养大鼠海马神经元NMDA受体作用位点的研究

目的：研究新型tacrine双联体bis（7）-tacrine对NMDA受体的作用位点.方法：大鼠海马神经元原代培养,全细胞膜片钳记录培养大鼠海马神经元上NMDA激活电流变化.结果：细胞外液pH值从8.1改变到6.7,在细胞外液中加入二硫苏糖醇（2 mmol/L）、精胺（10 mol/L）、镁离子（50～500 mol...     

Effect of Nano red elemental selenium on GPx activity of broiler chick kidney cells<Emphasis Type="Italic">in vitro</Emphasis>

A new selenium source, Nano red elemental selenium (Nano-Se) was used to study the effect on the GPx activity of broiler chick kidney cells (BCKC)in vitro, Sodium selenite (Na₂SeO₃) and seleno-1-methionine (Se-Met) were used as the controls. The results showed that the effects of three kinds of Se forms on the GPx activity of BCKC were accordant (p>0.05) compared with each other at 0.01. 0.05 and 0.10 μmol/L Se concentrations treatments. In the range of 0.00–0.10 μmol/L Se concentrations, the GPx activity increased with elevation of Se concentrations in medium. For the three kinds of Se forms, the GPx activity reached the climax at 0.10 μmol/L Se concentration. At 0.20 and 0.30 μmol/L Se concentrations, the influnces of three kinds of Se forms were not accordant with one another. For Nano-Se, the GPx activity at 0.20 and 0.30 μmoi/L Se concentrations remained the same as that at 0.10 μmol/L Se concentration treatment. For Se-Met, the GPx activity at 0.20 μmol/L Se concentration treatment remained the same with 0.10 μmol/L treatment; the GPx activity at 0.30 μmol/L Se concentration treatment was declined significantly (p<0.05) compared with 0.10 or 0.20 μmol/L treatment. For Na₂SeO₃, the GPx activity falled gradually with Se concentration increasing from 0.10 μmol/L to 0.30 μmol/L, and at 0.30 μmol/L Se concentration treatment, the GPx activity was less than the original of BCKC. The results implicated, on the GPx activity of BCKCin vitro, the ranking of width range of the most suitable Se concentration for nutrition curve of the three Se formes is Nano-Se>Se-Met>Na₂SeO₃. Foundation item: Supported by the Key Item of the Science and Technology Bureau of Zhejiang Province (021122680) Biography: Xu Bao-hua (1966-), male, Associate professor, Ph. D. research direction: nanobiology and animal nutrition.     

Cloning,expression and binding specificity analysis of odorant binding protein 3 of the lucerne plant bug, <Emphasis Type="Italic">Adelphocoris lineolatus</Emphasis> (Goeze)

Credible evidence shows that odorant binding proteins (OBPs) are required for insect olfaction perception and play a key role in transporting hydrophobic odorants across the sensillum lymph to the olfactory receptors (ORs). In the present study, a novel OBP (AlinOBP3) gene from the lucerne plant bug, Adelphocoris lineolatus, was cloned and expressed. The expression pattern of AlinOBP3 was evaluated by qPCR, which indicated that AlinOBP3 was dominantly expressed in antennae. The binding properties of AlinOBP3 with 9 cotton volatiles and 5 sex pheromone analogs were measured by fluorescence competitive binding assays with the fluorescence probe 1-NPN. The results revealed that of 9 cotton volatiles, Myrcene, β-Ocimene and α-Phellandrene can bind with AlinOBP3. α-Phellandrene especially bound to AlinOBP3 with a high binding affinity, with a dissociation constant of 56.68 μmol/L. Of the 5 sex pheromone analogs, Hexyl butyrate had the strongest binding affinity with AlinOBP3, with a dissociation constant as 59.53 μmol/L. Butyl butyrate, trans-2-Hexenyl butyrate and Ethyl butyrate had medium binding affinities with AlinOBP3, with dissociation constants of 227.39, 108.77 and 143.47 μmol/L, respectively. The results suggest that AlinOBP3 might be a pheromone binding protein (PBP) with a dual-function for the perception of sex pheromones and plant volatiles.     

Effect of CO<Subscript>2</Subscript> concentrations on the activity of photosynthetic CO<Subscript>2</Subscript> fixation and extracelluar carbonic anhydrase in the marine diatom <Emphasis Type="Italic">Skeletonema costatum</Emphasis>

The growth and activity of photosynthetic CO2 uptake and extracellular carbonic anhydrase (CAext) of the marine diatom Skeletonema costatum were investigated while cultured at different levels of CO2 in order to see its physio-logical response to different CO2 concentrations under either a low (30 靘ol·m-2·s-1) or high (210 靘ol·m-2·s-1) irradiance. The changes in CO2 concentrations (4—31 靘ol/L) affected the growth and net photosynthesis to a greater extent under the low than under the high light re-gime. CAext was detected in the cells grown at 4 mol/L CO2 but not at 31 and 12 靘ol/L CO2, with its activity being about 2.5-fold higher at the high than at the low irradiance. Photo- synthetic CO2 affinity (1/ K1/2(CO2)) of the cells de-creased with increased CO2 concentrations in culture. The cells cultured under the high-light show significantly higher photosynthetic CO2 affinity than those grown at the low-light level. It is concluded that the regulations of CAext activity and photosynthetic CO2 affinity are dependent not only on CO2 concentration but also on light availability, and that the de-velopment of higher CAext activity and CO2 affinity under higher light level could sufficiently support the photosyn-thetic demand for CO2 even at low level of CO2.     

Bis(7)-tacrine ameliorates cognitive impairment caused by cerebral hypoperfusion in rats

The effects ofbis(7)-tacrine, a novel acetylcholinesterase inhibitor, on cognitive impairment and neuronal degeneration induced by permanent ligation of bilateral common carotid arteries (2VO) were investigated using the Morris water maze in rats. Once daily oral administration ofbis(7)-tacrine (0.025, 0.05 and 0.1 mg·kg⁻¹) was started 14 days after 2VO operation. Over a three-week treatmentbis(7)-tacrine effectively reversed 2VO-induced spatial memory deficits in a dose-dependent fashion. Furthermore, histological observation showedbis(7)-tacrine decreased the neuropathological changes in the 2VO rats brain. These results suggest thatbis(7)-tacrine has therapeutic potential for the treatment of dementia caused by chronic cerebral hypoperfusion. Foundation item: Supported by the National Natural Science Foundation of China (No: 30170335) Biography: Peng Feng-fang(1966-), female Master, research direction: Alzheimer's disease and ischemia     

Pinacidil,a K<Subscript>atp</Subscript> channel opener,identified as a novel agonist for TRPA1

The transient receptor potential Ankyrin 1(TRPA1) cation channel is activated by various pungent and irritant compounds,and it also mediates the perception of noxious cold.Identification of different agonists for this channel is important for understanding its activation mechanism.Therefore,a screen for novel TRPA1 agonists was performed using an agonist-induced calcium influx assay.Out of 90 compounds screened,pinacidil was identified as a novel agonist for this channel.Pinacidil is a known opener of the K atp channel,for which it has an EC50 value of 1-3 μmol/L.In comparison,the EC50 value of pinacidil for TRPA1 is relatively high(260 μmol/L).Recombinant HEK-TRPA1 cells did not respond to P1075,another K atp channel opener,suggesting that the effect of pinacidil on TRPA1 was highly specific.Further studies revealed that the agonist activity of pinacidil could be blocked by the TRP channel inhibitors,ruthenium red and HC-030031.Using glutathione(GSH) and site-specific mutagenesis,we demonstrated that pinacidil could activate TRPA1 by covalent modification of the critical amino acids C619,C639 and C663 in the N-terminus of TRPA1.     

Involvement of Oxidative Stress and Down-Regulation of Bcl-2 in Arachidonic Acid-Induced Apoptosis in HUVECs

0IntroductionAraacicdhi,diosn fiocu ancidd p(reAdAo)m i,naanntelsys eantt itahle p sonl-y2u npsoastiutiroante doff actetl-ylular phospholipids . Normal free AAconcentrationin humanblood ranges from5 .8μmol/Lto 49 .3μmol/L[1].It is re-leased mostlythroughactivation of phospholipase A2by physi-ological and pathological sti muli[2]. Free AAcan be metabo-lizedinto various eicosanoids via specific enzymes such as cy-clooxygenases ( COX) , lipoxygenase and cytochromesP-450[3]. During AA met…     

Bis(7)-Tacrine Ameliorates Cognitive Impairment Caused by Cerebral Hypoperfusion in Rats

The effects of bis(7)-tacrine, a novel acetylcholinesterase inhibitor, on cognitive impairment and neuronal degeneration induced by permanent ligation of bilateral common carotid arteries (2VO) were investigated using the Morris water maze in rats. Once daily oral administration of bis(7)-tacrine (0.025, 0.05 and 0.1 mg\5kg-1) was started 14 days after 2VO operation. Over a three-week treatment bis(7)-tacrine effectively reversed 2VO-induced spatial memory deficits in a dose-dependent fashion. Furthermore, histological observation showed bis(7)-tacrine decreased the neuropathological changes in the 2VO rats brain. These results suggest that bis(7)-tacrine has therapeutic potential for the treatment of dementia caused by chronic cerebral hypoperfusion.     

Synergistic effect of trace elements and flavonoids from Epimedium koreanum Nakai on primary osteoblasts

Several trace elements, particularly, manganese （Mn） and zinc （Zn）, are essential in bone metabolism as cofactors for specific enzymes. It has been reported that there exists the relationship between osteoporosis and trace element-deficiency and the efficacy of Ca, Mn and Zn supplementation on spinal bone mineral density in postmenopausal women. Traditional Chinese medicines （TCM）, such as Herba epimedii, were proved to be effective for prevention of osteoprosis in vivo; however, the efficacy of the main constituents and/or crude extract was not ideal in vitro, which suggested that they may work in another way. The purpose of the present study was to examine whether the combination of icariin and total flavonoids （TF） from Herba epimedii with mineral elements, which were abundant in Herba epimedii, would have a more beneficial effect on the viability and differentiation of primary osteoblasts than either agent alone, and to analyze the dada for a possible synergistic, additive or antagonistic effect. The combinations of 10 μmol/L Zn, Ca and Mn with icariin and total flavonoids greatly improved the cell viability and meanwhile dramatically enhanced the alkaline phosphatase activity as compared to each agent alone. On the other hand, an increased cell growth inhibition was also observed by combining 0.1 μmol/L, 1 pmol/L Zn with 10μmol/L icariin, and 10 μmol/L Mn with 0.06 μg/mL total flavonoids. Meanwhile a decreased alkaline phosphatase activity was also found in several icariin-Zn/Mn and total flavonoids-Zn/Ca/Mn combinations. These results suggested that mineral elements （Zn, Ca, Mn） greatly enhanced the efficacy of icariin and total flavonoids from Herba epimedii on the viability and differentiation of primary osteoblasts by certain combinations.     

Effect of Cl- on photosynthetic bicarbonate uptake in two cyanobacteria Microcystis aeruginosa and Synechocystis PCC6803

Photosynthetic inorganic carbon utilization was investigated in two cyanobacteria Microcystis aeruginosa and Synechocystis PCC6803 grown in standing culture. Photosynthetic rates for the two algae reached about 10 times the theoretical CO₂ supply rate at low dissolved inorganic carbon (DIC) of 100 μmol/L, and the rates were unaffected by the addition of 20 mmol/L Na⁺, indicating that the two algae possessed Na+-independent HCO^-₃ utilization for photosynthesis under low DIC. Their photo- synthetic rates at low DIC were inhibited by higher Cl^- and the degrees of inhibition were increased with the rise of CI^- concentration, and in the presence of Diphenylamine-2-carboxylate (DPC), a reported Cl^- channel inhibitor, the rates decreased by 74%－82%, implying that putative DPC-sensitive Cl^- channels participate in Na+-independent HCO₃^- uptake for photosynthesis. The experiment of intracellular 14C-DIC accumulation for photosynthesis showed that internal DIC pools decreased by about 80% with 200 μmol/L DPC and by 64%－70% with 100 mmol/L Cl^-. The experiment of chlorophyll a fluorescence quenching showed that initial rates and extents of fluorescence quenching obviously decreased with 200 μmol/L DPC or 100 mmol/L Cl^-. The two experiments gave further evidence that putative DPC-sen- sitive Cl^- channels participate in Na+-independent HCO^-₃ uptake for photosynthesis in the two algae grown in standing culture.     

Modification of TiO2 nanotubes arrays by CdS and their photoelectrocatalytic hydrogen generation properties

In order to realize hydrogen generation under visible light, novel CdS/TiO2 nanotubes arrays are developed by electrochemical anodizaUon of Ti in 0.15 mol/L NHTF ＋ 0.08 mol/L H2C2O4 electrolyte. The diameter of the nanotube is 80-100 nm and the length is approximately 550 nm. The CdS nano-particles are deposited on the TiO2 nanotubes arrays by chemical bath deposition （CBD） in the ammonia-thiourea system. A 300 W Xe lamp is used as the light source, CdS/TiO2 nanotube arrays are used as the photoanode with the application of 1.0 V bath voltage, and 0.1 mol/L Na2S ＋ 0.04 mol/L Na2SO3 solution is used as the electrolyte, then the rate of photoelectrocatalytic hydrogen generation is 245.4 μL/（h·cm^2）. This opens new perspectives for photoelectrocatalytic hydrogen generation by using CdS/TiO2 nanotubes arrays.     

Organic media enhanced spectrofluorimetric determination of L-ascorbic acid

A novel spectrofluorimetric method for the determination of L-ascorbic acid is proposed. It is based on the inhibition of L-ascorbic acid on the formation of 2,3-diaminophenazine, which is an oxidation product ofo-phenylenediamine catalyzed by laccase. The fluorescence (at λ_cx/λ_cm=464 nm /530 nnm) was enhanced strongly in the presence of organic media. The mechanism ofo-phenylenediamine oxidation reaction catalyzed by laccase in the presence of L-ascorbic acid is discussed. L-ascorbic acid is determined in the ethanol, 1,4-dioxane and acetone over the linear range of 4.0×10⁻⁷≈1.2×10⁻⁴ mol/L, 4.0×10⁻⁷≈8.0×10⁻⁵ mol/L and 4.0×10⁻⁷≈1.0×10⁻⁴ mol/L with a detection limit of 1.20×10⁻⁵ mol/L, 1.19×10⁻⁵ mol/L and 1.24×10⁻⁸ mol/L, respectively. The method has been successfully applied to the simple and rapid determination of L-ascorbic acid in pharmaceuticals and milk powder. Supported by National Natural Science Foundation of China and the Special Funds of State Education Committe for Doctorate Research Huang Zuyun: born in Aug. 1963. Ph.D     

An acetylcholinesterase inhibitor, 3-benzidino-5-methyl-6-phenylpyridazine, blocking outward potassium currents in acutely isolated rat hippocampal pyramidal neurons

3-benzidino-5-methyl-6-phenylpyridazine (BMP) inhibited electric eel acetylcholinesterase (AChE), with IC50 being 0.58 μmol·L-1. As an AChE inhibitor, the effects of BMP on delayed rectifier potassium current (IK(DR)) and transient outward potassium current (IK(A)) in acutely isolated rat hippocampal pyramidal neurons were studied using the whole cell patch-clamp technique. BMP (0.1―50 μmol·L-1) inhibited IK(DR) and IK(A) in a concentration-dependent and voltage-independent manner. The IC50 value for the bl...     

Circular dichroism spectral studies on the recombinant human neuroglobin

Neuroglobin (NGB) is a newly discov- ered member of the hemoglobin superfamily that is primarily expressed in the brain of humans and other vertebrates. The effects of protein concentration, solvent, pH and temperature on the secondary structure of NGB were investigated by employing far UV circular dichroism (CD) spectroscopy. The results show that NGB exists mainly in α-helix form when its concentration is less than 10 μmol/L. However, its α-helix content decreases with the increase of con- centration in the range of 10―40 μmol/L and remains unchanged when the concentration is higher than 40 μmol/L, which suggest that NGBs form intermolecular disulfide bond and aggregate in higher concentration. The α-helix content of NGB in methanol and ethanol is a little higher than that in water, indicating a higher stability of NGB in these solvents. NGB loses its α-helical secondary structure in either acidic or alka- line solution to some extent. Although increased temperature destabilizes the α-helices of NGB, over 16% of α-helices can be kept at 110°C. Therefore, NGB is a protein with hyperthermal stability.     

Electrocatalytic oxidation of H<Subscript>2</Subscript>O<Subscript>2</Subscript> at a carbon paste electrode modified with a nickel (II)-5, 11, 17, 23-tetra-tert-butyl-25, 27-bis(Diethylcarbamoylmethoxy) calix[4] arene complex and its application

Electrochemical behavior of a carbon paste electrode (CPE) modified with nickel(II)-5, 11, 17, 23-tetra-tert-butyl-25, 27-bis(diethylcarbamoylmethoxy) calix[4]arene (Ni (II)-L) complex and its electrocatalytic activity towards the oxidation of hydrogen peroxide were investigated by cyclic voltammetric technique in a 5.0×10⁻² mol/L NaClO₄+1.0×10⁻³ mol/L NaOH solution. It was found that Ni(II)-L acts as an effective catalyst for the oxidation of hydrogen peroxide. The modified electrode exhibited a linear response over a hydrogen peroxide concentrations in the range of 2.0×10⁻⁵−1.0×10⁻⁴ mol/L with a detection limit as low as 1.0×10⁻⁶ mol/L. The relative standard deviation was 3.5% for 5 successive determinations of H₂O₂ at 1.0×10⁻⁵ mol/L. The modified electrode was used successfully in rainwater analysis. Foundation item: Supported by the Natural Science Foundation of Hubei Province (98J040). Biography: Li Chun ya(1972-), Ph. D. candidate, research direction: electroanalysis and electrosynthesis.     

Overinhibition of Mitogen-Activated Protein Kinase Inducing Tau Hyperphosphorylation

To reveal the relationship between mitogen-activated protein kinase (MAPK) and tau phosphorylation, we used different concentration of PD98059, an inhibitor of MEK (MAPK kinase), to treat mice neuroblastma (N2a) cell line for 6 h. It showed that the activity of MAPK decreased in a dose-dependent manner. But Western blot and immunofluorescence revealed that just when the cells were treated with 16μmol/L PD98059, tau was hyperphosphorylated at Ser396/404 and Ser199/202 sites. We obtained the conclusion that overinhibited MAPK induced tau hyperphosphorylation at Ser396/404 and Ser199/202 sites.     

铂微粒修饰固体石蜡碳糊电极对过氧化氢的电催化特征及分析应用

研究铂微粒修饰的固体石蜡碳糊电极(Pt-SCPE)对H2O2的电催化行为。在pH值4.6的柠檬酸盐-0.03 mol/L硝酸钾溶液中,用该电极H2O2在+0.18V(vs．SCE)可产生灵敏的还原电流,线性范围为1.0×10-1 mol/L～3.0×10-7 mol/L,检出限为7.5×10-8 mol/L。Pt-SCPE与液体石蜡碳糊电极比较,在峰电位、线性响应范围、检出限方面有更好的结果。用该电极测定样品中的过氧化氢,获得满意结果。     

Research at channel level on the effect of LaCl3 on Radish (Raphanus satirus L.) vacuolar membrane

We recorded slow vacuolar (SV-type) channel currents of Radish vacuoles successfully for the first time by using the whole-vacuolar patch-clamp recording mode. SV- type currents would increase and threshold potentials of activation would shift towards more negative values with the increase of concentrations of cytosolic Ca²⁺. When 2.5 mmol/ L LaCl₃ and 4 mmol/L EGTA were added to bath solutions, SV-type currents were suppressed remarkably. Then adding LaCl₃ with different concentrations to pipette solutions, we found that LaCl₃ with higher concentrations (＞4 ´ 10^-7 mol/L) had a strong inhibitory effect on SV-type currents, while LaCl₃ with lower concentrations (≤4 ´ 10^-7 mol/L) promoted channel currents. This promoting effect provides an important basis at channel level for researching further the effects of rare earth on physiological activities of plants and the production-increase effects of rare earth fertilizers on crops.     

Study of the voltammetric behavior of heme and its direct analysis application

The electroanalytical method of ferriheme was studied by linear sweep voltammetry in medium of 0.05 mol/L Tris+0.05 mol/L NH₃−NH₄Cl buffer at hang mercury drop electrodes (HMDE). Heme exhibits two pair reversible redox peaks and one irreversible peak. The cathodic peak potentials are−;0.236 V, −0.422 V and −1.408 V respectively. The first and the third peaks can be used for directly quantitative determination of heme concentrations. The peak currents are good linear relationship with heme concentration in ranges of 3×10⁻⁶–6×10⁻⁵ mol/L and 3×10⁻⁷–1.5×10⁻⁵ mol/L respectively. Supported by the National Natural Science Foundation of China Lju Huihong: born in 1965, Lecturer. Visiting scholar from Xiangyang Teacher’s College, Hubei, 441053