Atomic force microscopic observation on substructure of pollen exine inCedrus deodara andMetasequoia glyptostroboides

The substructure of pollen exine inCedrus deodara (Roxb.) Loud. andMetasequoia glyptostroboides Hu et Cheng has been examined with an atomic force microscope (AFM). The results indicate that the exine substructure units containing sporopollenin in two species are similar in shape, which are granular, but slightly different in size. InCedrus the substructure unit of pollen exine appears to be 56–99 nm long and 42–74 nm wide, while inMetasequoia it appears to be 81–118 nm long and 43–98 nm wide. It has been observed that the subunits of pollen exine inCedrus arranged tightly to form short-rod-like or spheroidal pollen exine units, several or more than ten of which formed an island-like structure. There are various spaces among these island-like structures which are interconnected to occupy the entire pollen exine. InMetasequoia, the subunits of pollen exine also arranged tightly with a distribution tendency of cluster of 3–10, however, no obvious boundary exists among these clusters. From our results, it is concluded that there is no tendency of helical arrangement for the subunits of pollen exine inCedrus andMetasequoia, and the results support Southworth’ view that subunits of pollen exine are granular shape in lattice structure.     

Nitrogen-fixing inhibition by KCl in a blue-green algaAnabaena cylindrica

Growing in the medium containing 0.10 mol/L KCl for three days, the nitrogen-fixing activity ofAnabaena cylindrica decreased by 95%. Extent of the inhibition was enchanced with increased KCl concentrations. The biggest decrease of nitrogen-fixing activity was between 0.02–0.05 mol/L KCl. The same decline on growth was between 0.05–0.10 mol/L KCl. The longer treating-time was, the lower nitrogen-fixing activity was. A short time of KCl-treatment (for 4 h) resulted in 66.4% decrease of the activity. Under effecting of KCl for three days, the frequency of heterocyst differentiation decreased from 7.2% to 2.1%. Supported by the National Natural Science Foundation of China Yi Ping: born in 1974, Graduate student     

Regenerating plants from cryopreserved adventitious buds of haploids in rice

A large number of adventitious buds were induced fromin vitro cultured young inflorescences of haploids of rice. Having been subcultured on solidified subculture media at 26°C for 7 days, the adventitious buds were loaded into 1.8 mL plastic cryotubes with cryoprotectant and kept on ice for 45–60 min. After cooled at a rate of 1.0°C/min down to −40°C, the samples, were kept in liquid nitrogen. The adventitious buds which have been cryopreserved for about 30 days were thawed rapidly in 38–40°C water and then plated on solidified MS medium containing 3% sucrose, 0.5 mg/L NAA and 2.0 mg/L kinetin. After plated, 23%–32% of adventitious buds resumed growth and 15%–22% regenerated plantlets. The results of this work indicated that the adventitious buds derived fromin vitro cultured young inflorescences is a critical factor for the success and subculturing adventitious buds on MS medium containing 3% sucrose and 4% sorbitol or 20% potato extract is essential to the procedure. The effective cryoprotectant is 10% DMSO (dimethyl sulfoximide)+0.5 mol/L sorbitol. Supported by the Natural Science Foundation of Hubei Province Zhang Zhihong: born in 1963, Lecturer     

HPLC determination of ascorbic acid using luminol-Cu (II) chemiluminescence-application to the analysis of Juice beverage

The chemiluminescence (CL) of luminol-Cu (II) was applied to HPLC determination of ascorbic acid, which was separated by a C₁₈ reverse-phase column with a mobile phase of 0.25 mol/L HAc. The eluted ascorbic acid was mixed with 0.3 mmol/L luminol and 0.05 mol/L CuSO₄. The light emission from the reaction of Cu(II) oxidized ascorbic acid and luminol was detected by a modified luminometer. The detection limit was 3.6×10⁻⁶ mol/L for ascorbic acid at aS/N ratio of 3, and the linear calibration range was 2×10⁻⁴–2×10⁻³ mol/L. The relative standard deviation for 5 replicate injections of 1×10⁻³ mol/L ascorbic acid was calculated as 4.3%. The method was successfully applied to determination of ascorbic acid in juice beverage. Foundation item: National Natural Science Foundation of China (29605001) Biography: Wu Feng-wu(1963-), male, research direction: analytical chemistry.     

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.     

A novel method for thermodynamic study on binding of copper ion with Alzheimer’s amyliod β peptide

The interaction of Cu²⁺ with the first 16 residues of the Alzheimer's amyliod β peptide, Aβ (1-16), was studied by employing isothermal titration calorimetry at pH 7.2 and 37℃ in aqueous solution. The Gholamreza Rezaei Behbehani (GRB) solvation model was used to reproduce the enthalpies of Cu²⁺+ Aβ(1-16) interaction over the whole Cu²⁺ concentrations. The binding parameters recovered from the solvation model were attributed to the structural change of Aβ (1-16) due to the metal ion interaction. It was found that there is a set of two identical and non interacting binding sites for Cu²⁺ ions. The molar enthalpy of binding is ΔH=27.895 kJ/mol. The association binding constants are 1.895 μm^-1 and 1.891 μm^-1 for the first and second binding sites respectively.     

Adrenomedullin inhibits the endothelin production induced by urotensin II in rat vascular smooth muscle cells

The aim of this study was to observe the effects of adrenomedullin (ADM) on endothelin (ET) production induced by urotensin Ⅱ (UⅡ) in rat vascular smooth muscle cells (VSMCs). Cultured VSMCs which were incubated with UⅡ (10^-8 mol/L) and with various concentrations of ADM were used to measure the VSMCs ³H-TdR incorpora- tion, the activity of extracellular signal-regulated kinase (ERK), the amount of ET mRNA and ET production in VSMCs. In this work we found that incubation with UⅡ(10^-8 mol/L) increased obviously the amount of ET mRNA in VSMCs and ET production in medium, however, coincubation with ADM (10^-10—10^-8 mol/L) and UⅡ(10^-8 mol/L) reduced the amount of ET mRNA by 15%, 24% and 45% (P< 0.01) respectively, compared with UⅡ alone. The content of ET in medium was 14.13, 11.38 and 11.00 pg/mL. ADM alone (10^-8 mol/L) had no effect on ET production in VSMCs. UⅡ (10^-8 mol/L) promoted the 3H-TdR incorpo- ration and activity of ERK in VSMCs. ADM inhibited VSMCs 3H-TdR incorporation and activation of ERK in a concentration-dependent manner. Compared with UⅡ group, after coincubation with ADM (10^-10—10^-8 mol/L) and UⅡ (10^-8 mol/L) the VSMCs 3H-TdR incorporation was decreased by 7% (P > 0.05), 32% (P < 0.05) and 41% (P < 0.01), respectively, and the activity of ERK was decreased by 24% (P > 0.05), 32% (P < 0.05) and 36% (P < 0.05), re- spectively, in a concentration-dependent manner. The results show that in cultured VSMCs ADM inhibits ET mRNA expression, ET production and proliferation stimulated by UⅡ, and that inhibitory effect of ADM on UⅡ bioaction could be mediated through inhibiting MAPK pathway.     

Surface Dissociation Properties of Short Chain Carboxyl Mercaptan Self-Assembled Monolayers by Impedance Titration

Impedance titration was used to determine the surface dissociation characteristics of short-chain carboxyl mercaptan self-assembled monolayers （SAMs）. Based on the change of the cyclic voltammetric peak current and the charge-transfer resistance, which was related to pH value of the solution, the surface pKa of mercaptoacetic acid（MA）, 3-mercaptopropionic acid（MPA） and ω-mercaptohexanic acid（MHA） self-assembled membranes, with ionic strength being 0.1 mol/L, were determined to be 5.20, 4.80, 7.40, respectively. In addition, factors such as time needed for assembling, structure of monolayers and ionic strength, which effected the surface pKa, were studied as well. Such surface pKa shifts were sufficiently explained by interactions between interfacial molecules and hydrophobicity.     

蜘蛛抱蛋属花粉扫描电镜研究

为进一步研究蜘蛛抱蛋属植物的起源,演化及自然分类,用扫描电子显微镜观察蜘蛛抱蛋属（Aspidistra)的棒芯蜘蛛抱蛋（A.claviformis),丛生蜘蛛抱蛋（A.caespitosa)、峨边蜘蛛抱蛋（A.ebianensis)、辐花蜘蛛抱蛋(A.subrotata)、棕叶草（A.oblanceifolia)、广东蜘蛛抱蛋（A.lurida)、乐山蜘蛛抱蛋（A.leshanensis)、海南蜘蛛抱蛋（A.hainanensis)、大花蜘蛛抱蛋（A.tonkinensis)、罗甸蜘蛛抱蛋（A.luodianensis)的花粉形态,每种植物采发育正常含包待放的花1－3朵,取花粉用艾特曼醋酸法处理后,CO2临界点干燥,导电胶贴于样台,真空喷金镀膜,置于扫描电子显微镜下观察、测量及照相。详细地描述各种花粉的外部形态。10种蜘蛛抱蛋属植物花粉均为近球形或球形,无萌发孔,外壁纹饰可分为：皱波状纹饰和小芽孢状纹饰。不同种植物花粉外壁纹饰存在一定差异,二类型花粉外壁纹存在一定的演化关系。     

Research of Inclusion Complexation of 4,4＇-Thiodiphenol Effected by β-Cyclodextrin in Aqueous Solutions and the Effect on Photodegradation

The inclusion behavior of 4, 4＇ -Thiodiphenol （TDP）, a typical bisphenol and endocrine disruptor, reacts with β-cyclodextrin （β-CD） in aqueous solutions has been investigated by means of UV absorption spectrum and quantum-chemical calculation with Gaussian 98 software. The results show that the inclusion behavior of TDP is quite different in acidic solutions （pH 5.9） from that in alkaline solutions （pH 10.0）. This behavior difference is attributed to the different formula structures in aqueous solutions at acidic and alkaline pH values that are demonstrated by quantum- chemical modeling and calculation. TDP forms a 1 ： 1 fl-CD inclusion complex in aqueous solutions. The equiiibrium constant K was calculated to be 553.49 L/mol at pH 5.9 and 1 318.20 L/mol at pH 10.0 respectively for the inclusion complex reaction by using the modified Benesi-Heldbrand equation. After inclusion TDP＇s structure is changed especially at the inclusion part with the bond order becoming larger, which results in inhibitive photodegradation during direct photooxidation and H2O2 assisted photooxidation.     

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.     

发酵木糖高产乙醇树干毕赤酵母菌株的Co60诱变选育

【目的】选育能够利用木糖高产乙醇的酵母菌株。【方法】采用Co60诱变树干毕赤酵母(Pichia stipitis),筛选乙醇产量高的突变菌株,并对原始菌株、突变菌株的生长发酵特性和两个菌株对高浓度木糖、乙醇的耐受性进行比较。【结果】在YPX培养基上筛选获得1株能够高效发酵木糖的突变菌株1K-9。该菌株在50mL 15%FM发酵84h,发酵液乙醇含量最高达(51.034±0.112)g/L,比原始菌株提高10.05%;在500mL 15%FM发酵96h,乙醇含量最高达(51.390±0.119)g/L;在500mL 20%FM发酵156h,乙醇含量最高达(52.496±0.513)g/L。菌株1K-9在HSM培养基或含4%~5%乙醇的YPX培养基中生长良好,在含6%~7%乙醇的YPX培养基中生长缓慢。【结论】Co60诱变对于树干毕赤酵母(Pichia stipitis)菌株是有效的,能选育出木糖高产乙醇酵母菌株1K-9。     

Bis(7)-Tacrine, a promising anti-Alzheimer's agent, attenuates glutamate-induced cell injury in primary cultured cerebrocortical neurons of rats

The effects of bis(7)-tacrine, a novel dimeric acetylcholinesterase (AChE) inhibitor, on glutamate-induced cell injury were investigated in primary cerebral cortical neurons of rats. Exposure of cultured neurons (12 days after plating) to 0.5 mmol/L glutamate for 30 min resulted in significant cell damage. Pretreatment with bis(7)-tacrine (0.03–1.0 μmol/L) reduced the glutamate-induced neurotoxicity in a concentration dependent manner and the maximal response was seen at 1 μmol/L with approximately, 30% protection. A receptor binding assay showed that bis(7)-tacrine can completely displace MK-801 binding to rat cortical membrane with an IC₅₀ of 0.57 μmol/L. These findings suggest that bis(7)-tacrine can directly interact with N-methyl-D-aspartate receptor channel complex, which may contribute to the inhibitor's protective effects against glutamate-induced excitotoxicity. Thus, it is possible that anti-glutamate/anti-AChE synergism is responsible for potentially better Alzheimer's therapy of bis(7)-tacrine relative to tacrine. Biography: Zhang Bai-fang (1974-), female, Ph. D. candidate, research direction: Alzheimer's disease and ischemia     

Pollen Morphology of Artemisia L. and Its Systematic Significance

The pollen of 57 species representing 7 sections of Artemisia was examined hy light microscope and scanning microscope for the purpose to elucidate the systematic relationships of the genus. The examination indicates that the pollen grains present high concordance which is characterized by the globular or the appruximate shape. 3-lobed circular from polar view while granular or ellipse sphere from tropical plate with tricolporate structure. The exinc of pollen has an obvious double structure of inner and outer layers (including the tecturn and columnar layers) where the outer is thicker than the inner and the sculpture of exine degenerates into small spinules easily distinguishable from other genera of the Compositae. Based on cxine sculpture, the pollen grains of Artemisia can be divided into two types: Myriantha type and Mongolica type, which possesses 4 subtypes according to the distribution forms of spinulc: (1) Sacrorum subtype, (2) Oligocarpa subtype, (3) Lavandulaefolia subtype. (4) Anomale subtype.     

Development of an analytical method to determine phenolic endocrine disrupting chemicals in sewage and sludge by GC/MS

An analytical method has been developed to determine hisphenoi A and five degradation products of alkyiphenoi ethoxylates in sewage and sludge. The parameters of accelerated solvent extraction （ASE） were optimized. C18 cartridge was used to clean up the sewage and the extract of sludge. The sensitivity of the mass detector was improved by adjusting the voltage of the lens manually. The instrumental detection limit was 3.2-17.8pg. The method detection limits were 2.5-11.4 ng/L and 0.3-2.1ng/g for sewage and sludge analysis respectively. The recovery and relative standard deviation for sewage analysis were 79.2%-111.4% and 6.9% - 12.6%; for sludge analysis were 68.5% - 114.0% and 8.9%-16.0%. Finally this method was applied to determining the phenolic endocrine disrupting chemicals in sewage and sludge from a sewage treatment plant in Beijing.     

井冈山地区铁皮石斛的繁殖生物学研究

通过野外实验观察和人工控制套袋等方法,从铁皮石斛的花部基本特征、开花动态、花粉活力、柱头可授性、杂交指数、套袋实验等方面对其繁殖生物学进行了研究。结果显示:(1)井冈山地区铁皮石斛4～6月开花,种群花期近60 d,多数花(50%)在约30 d内开放,果实一般需要180 d左右才能成熟。(2)铁皮石斛花粉活力开花后24 h即可达到(62.67±6.43)%,至开花第三天,活力可达到90%,柱头可授性达到最高,至第5天柱头仍保持高可授性。(3)套袋实验表明,部分自交亲和,异交,需要传粉者。自然条件下未观察到井冈山铁皮石斛居群结实,人工自花授粉率为22.2%。该研究结果为保护和利用井冈山铁皮石斛种质资源提供了科学依据。     

Transformation of plant young proembryos by electroporation

It is first reported that plant young proembryos expressed exogenous reporter genes by electroporation. Young proembryos with 8–32 cells and globular proembryos with 250–400 cells could be isolated by enzymatic maceration combined with microdissection. After electroporation withGUS orGFP genes, the proembryos were cultured for 1–2 d in KM8p medium. At the field strength of electroporation 500–1500 V/cm, blue reaction of GUS or green fluorescence of GFP could be observed in the proembryos. The highest transient expression frequency of young proembryos (2.2%) was obtained at the field strength of 750 V/cm, whereas the highest frequency of globular proembryos (5.9%) was obtained at the field strength of 1 250 V/cm. Taking the proportion of transformed cells in the whole cells of proembryos as efficient transformation frequency, the efficient transformation frequency of the young proembryos was 7 times that of the globular proembryos.     

Formation of the inclusion complex of Orange II with β-Cyclodextrin and its photostability

Inclusion complex of Orange II with β-Cyclodextrin (β-CD) and the anti-photolysis effect under UV-light were investigated. The molar ratio of inclusion complex of β-Cyclodextrin and Orange II is 1∶1. The formation constantK = 1.236×10³ L/mol was determined by the UV and Fluorescence spectra respectively, which was quite in accordance with the calculation with a modified Benesi-Hildbrand equation. The inclusion complex was characterized by the IR spectra and the molar ratio of inclusion complex is 1∶1 too. The formation constantK = 1.266×10³ L/mol was determined by¹H NMR analysis and was nearly the same by UV and fluorescence spectra. The photocatalytic decolorization rate of Orange II solutions containing β-CD and TiO₂ was smaller by 51.9% than that of the Orange II solutions only containing TiO₂, while in the case of direct photolysis of Orange II solutions, β-CD can lower the photolysis rate by 48.1% under UV-light. This result indicates β-CD can inhibit the photolysis and photocatalytic decolorization of Orange II under UV-light. The β-CD inclusion complex was found to be persistent to UV-light photolysis. Foundation item: Supported by the National Natural Science Foundation of China (20177017) Biography: Lu Ping (1958), male. Associate professor, research direction: environmental photochemistry.     

Chemiluminescence determination of thiourea using tris (1,10-phenanthroline)-ruthenium (II) KMnO4 system

The light emission produced by thiourea in oxidation process by permanganate in acidic solution in the presence of Ru(phen) ₂ ^{3 +} is used to determine 1.0×10⁻⁵ −1.0×10⁻⁵ mol/L thiourea. The limit of detection is 9.8×10⁻⁹ mol/L and the relative standard deviation is 1.1% for a 1.0×10⁻⁵ mol/L thiourea solution (n=10). The method was applied satisfactorily to the determination of thiourea. Foundation item, Support by Ministry of Education, Wuhan University and Alumni Association of it Biography: Wu Feng-wu(1963-), Male, Ph. D, research direction: analytical chemistry.     

Characterization of Anabaena cylindrica Solution System Using Synchronous-Scan Fluorescence Spectroscopy

The characterization of the algaeAnabaena cylindrica solution with Fe (III) was investigated using fluorescence emission and synchronous-scan spectroscopy. The ranges of concentrations of algae and Fe (III) in aqueous solutions were 5.0×10⁸ 2.5×10⁸ cell/L and 1060 μmol/L. respectively. The effective characterization method used was synchronous-scan fluorescence spectroscopy (SFS). The wavelength difference (Δλ) of 90 nm, was maintained between excitation wavelength (λen) and emission wavelength (λen). The peak was observed at about (λex) (λem) 326 nm for synchronous scan fluorescence spectroscopy. The fluorescence quenching in system of algae Fe(III)-HA was studied using synchronous-scan spectroscopy for the first time, Fe(III) was clearly the effective quencher. The relationship betweenI I (quenching efficiency) andc (concentration of Fe (III) added) was, a linear correlation for the algae solution with Fe(III). Also, Aldrich humie acid (HA) was found to be an effective quencher. Foundation item: Supported by the National Natural Science Foundation of China (20177017) and the Scientific Research Foundation of Wuhan Environmental Protection Bureau Biography: LIU Xian li(1965), male. Ph.D. Associate professor, research direction: environmental chemistry.