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    25 July 2011, Volume 27 Issue 4
    Content
    Cover and Content of Chemical Research in Chinese Universities Vol.27 No.4(2011)
    2011, 27(4):  0. 
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    Articles
    Ionothermal Synthesis and Phase Transformation of Organic-inorganic Hybrid Neutral Zincophosphate Cluster [Zn(HPO4)(H2PO4)][C6H10N3O2]
    DONG Zhao-jun, YAN Yan, ZHENG Rong-feng, LIU Dan, LI Ji-yang, HAN Zhen-guo, ...
    2011, 27(4):  531-535. 
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    Ionothermal synthesis was used to prepare a novel amino acid containing hybrid zincophosphate monomer, [Zn(HPO4)(H2PO4)][C6H10N3O2](denoted as ZnPO-CJ58). The inorganic framework of [Zn(HPO4)(H2PO4)][C6H10N3O2] consists of 4-membered rings formed by ZnO3OHis and PO2(OH)2 tetrahedra. The HPO4 and amino acid moieties hang on the Zn center. Such a framework is stabilized by extensive multipoint hydrogen bonds involving the phosphate units and histidine molecules to form a pseudo-3D supramolecular structure. It is noteworthy that ZnPO-CJ58 is the first zinc phosphate cluster with amino acid acting as the ligand. It exhibits photoluminescence  excited at a wavelength of 220 nm. Interestingly, ZnPO-CJ58 can transform into a layered structure (C6H10N3O2)Zn2(HPO4)(PO4)·H2O(ZnPO-CJ36) through further reacting with water or zinc acetate dihydrate in water at 85 °C for 1 h. This work will be helpful for the synthesis of crystalline inorganic-organic hybrid materials with biofunctional molecules.
    Synthesis and Structure of an Organo-templated Oxouranium Phosphite
    YU Jian-guo* and ZHAO Yong-nan
    2011, 27(4):  536-539. 
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    A novel layered oxouranium phosphite open-structure [(C10H24N2)(H2O)][(UO2)2(HPO3)3](denoted as TJPU-8, TJPU=Tianjin Polytechnic University) was hydrothermally synthesized with isophorondiamine(IPDA) as the template. Single crystal structure refinement disclosed that TJPU-8 crystallized in the monoclinic space group P21c with cell parameters a=1.8346(9) nm, b=0.6734(1) nm, c=2.0639(0) nm, β=114.61(5)°, V=2.3182(8) nm3, Z=2. TJPU-8 was constructed by UO7 monomer, U2O12 dimer and HPO3 groups. The monomeric UO7 or dimeric U2O12 was connected by HPO3 groups to form two types of 4-ring chains. An undulating sheet was then generated by connecting these two types of chains via HPO3 bridges. The layer structure formed by stacking the sheets along the a direction via the strong H-bonds of organoamine cations and water molecules intercalated in the interlayer spaces with the framework oxygen atoms. Although a mixture of cis- and trans-IPDA was used, only cis-IPDA served as the template. A typical green light emission of TJPU-8 is observed when excited by 266 nm laser.
    Synthesis of CdTe/SiO2 Composites and Their Fluorescence Properties
    WANG Mei-ping*, LIN Jin-hui and SHAN Chuan-sheng
    2011, 27(4):  540-542. 
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    Thioglycolic acid(TGA)-stabilized CdTe nanocrystals(NCs) were prepared with sodium tellurite as   tellurium source, which avoids the cumbersome processes associated with H2Te or NaHTe sources. Fluorescent CdTe/SiO2 composites were synthesized by a sol-gel method without the exchange of surface ligands. The     phase structure of CdTe NCs was investigated by X-ray diffractometry. For comparison, some characterizations  were done for both the CdTe NCs and the composites. CdTe NCs and CdTe/SiO2 composites were characterized with TEM, digital camera and fluorescence spectrophotometer. The stability of CdTe NCs and the composites were    investigated in phosphate-buffered saline(PBS) buffer and the fluorescent properties of the composites were discussed in detail.
    Comparable Studies of Adsorption and Magnetic Properties of Ferrite MnFe2O4 Nanoparticles, Porous Bulks and Nanowires
    HOU Xiang-yu, FENG Jing*, LIU Xiao-han and ZHANG Mi-lin
    2011, 27(4):  543-546. 
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    The spinel ferrites MnFe2O4 nanowires were synthesized by hydrothermal route, porous MnFe2O4 and nanoparticles morphologies were synthesized by sol-gel method with egg white. The structures, morphologies, magnetic properties and adsorption properties of these obtained ferrites with different morphologies were studied contrastively. Results show that the obtained samples exhibit ferromagnetic properties. This realizes convenient magnetic separation from solution when they are used in the treatment of organic dyes wastewater. However, the contrastive studies show that the saturation magnetizations(Ms) of MnFe2O4 with different morphologies are different and the Ms follows the order: Ms(porous)<Ms(nanoparticles)<Ms(nanowires). In addition, the adsorptions of methylene blue(MB) onto these ferrites depend on ferrites’ morphologies seriously. The adsorption rate of MB on the porous MnFe2O4 is much higher than those onto the other two samples because the porous structure can provide high efficient mass transport through the pores.
    Fabrication of Graphene by Cleaving Graphite Chemically
    ZHAO Shu-hua, ZHAO Xiao-ting, FAN Hou-gang, YANG Li-li, ZHANG Yong-jun, ...
    2011, 27(4):  547-549. 
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    Graphite was chemically cleaved to graphene by Billups Reaction, and the morphologies and microstructures of graphene were characterized by SEM, Raman and AFM. The results show that the graphite was first functionalized by 1-iodododecane, which led to the cleavage of the graphene layer in the graphite. The second decoration cleaved the graphite further and graphene was obtained. The heights of the graphene layer were larger than 1 nm due to the organic decoration.
    Comparison of Different Cartridges of Solid Phase Extraction for Determination of Polyphenols in Tobacco by UPLC/MS/MS and Multivariate Analysis
    ZHANG Xia*, LIU Wei, XU Yong, YANG Liu, KONG Wei-song, RUI Xiao-dong, ...
    2011, 27(4):  550-556. 
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    The comparison of solid phase extraction(SPE) for the preconcentration and isolation of polyphenols in tobacco samples was carried out by ultra-high performance liquid chromatography/tandem mass spectrometry (UPLC/MS/MS) and multivariate analysis. Several adsorbing materials of SPE(C18, NH2, SAX and OASIS) were investigated. It was found that the C18 and OASIS cartridges can not only speed up the purification process, but also simplify the SPE operation. A UPLC/MS/MS was used for the determination of polyphenols in tobacco samples after purification. All analytes were separated and determined in 2 min. The limit of detection was 0.05 ng/mL. Cluster analysis(CA) and principal component analysis(PCA) were used for the analysis of 4 varieties(flue-cured tobacco, oriental tobacco, sun-cured tobacco and burley) in order to interpret the effect of planting and machining process on the concentration of polyphenols. The different types of tobacco samples could be easily clustered by CA. PCA on the chemical composition of tobacco resulted in two principal components(PCs) that take 84.2% of the total variance. The PCA and CA indicate that the polyphenols can be used for distinguishing tobacco types.
    Purification and Characterization of β-Glucosidase from a Newly Isolated Strain Tolypocladium cylindrosporum Syzx4
    ZHANG Yi-bo, YUAN Li-jia, CHEN Zhao-jie, FU Lu, LU Jia-hui, MENG Qing-fan, ...
    2011, 27(4):  557-561. 
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    A higher β-glucosidase-producing strain was isolated and identified as Tolypocladium cylindrosporum syzx4 based on its morphology and internal transcribed spacer(ITS) rDNA gene sequence. The present study is to ferment, purify and characterize a β-glucosidase from T. cylindrosporum gams. The enzyme was purified to homogeneity by sulfate precipitataion, diethylaminoethyl cellulose anion exchange chromatography and Sephadex G-100 gel filtration with a 9.47-fold increase in specific activity and a recovery of 12.27%. The molecular weight(Mw) of the β-glucosidase was estimated to be 58600 by SDS-PAGE, which is much lower than that of the enzyme from other fungi. The β-glucosidase shows high activity over a wide range of temperature from 35 °C to 70 °C and the optimum pH is approximately 2.4. Then the kinetic parameters Km(0.85 mmol/L) and vmax[85.23 mmol/(L·s)] of the β-glucosidase were studied, respectively with high affinity p-nitrophenyl-β-D-glucopyranoside(p-NPG) as the substrate, inhibition constant Ki(39.5 mmol/L) with the tolerance of glucose. Although β-glucosidases have been purified and characterized from several other sources, T. cylindrosporum β-glucosidase with the unique optimal pH and higher tolerance of glucose distinguished from others makes the β-glucosidase a potential application in simultaneous saccharification and fermentation(SSF).
    Screening and Identifying of Nephrotoxic Compounds in Lithospermum erythrorhizon Using Live-cell Fluorescence Imaging and Liquid Chromatography Coupled with Tandem Mass Spectrometry
    ZHAO Xiao-ping, JIN Ye-cheng, ZENG Xing, ZHANG Bo-li and ZHANG Yu-feng*
    2011, 27(4):  562-565. 
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    In order to identify the potential nephrotoxic compounds in traditional Chinese medicine Lithospermum erythrorhizon, it was separated into serial fractions according to their polarities. An in vitro method was utilized to determine the nephrotoxicity of these fractions with the help of fluorescence image analysis. As a result, the primary fraction A05 and its secondary fractions C06―C09 and C12―C14 were found to have significant toxicity to LLC- PK1 cell line, as determined by the survive rate less than 20% after they were treated with these fractions. These potential nephrotoxic fractions were further analyzed by multistage and high resolution mass spectrometry. The main compounds in these fractions were tentatively identified to be acetylshikonin, isobutyrylshikonin, β,β′-dimethyla- cryloylshikonin, and isovalerylshikonin, which may bring nephrotoxicity.
    Fe3O4 Magnetic Nanoparticles Modified Electrode as a Sensor for Determination of Nimesulide
    ZHANG Jin-lei, TAN Xue-cai*, ZHAO Dan-dan, TAN Sheng-wei, LIU Li, WANG Li, ...
    2011, 27(4):  566-569. 
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    A novel type of Fe3O4 nanoparticles modified glass carbon electrode(Fe3O4/GCE) was constructed and the electrochemical properties of N-(4-nitro-2-phenoxyphenyl)methanesulfonamide(nimesulide) were studied on the Fe3O4/GCE. In 0.4 mol/L HAc-NaAc buffer solution(pH=5.0), the electrode process of nimesulide was irreversible at bare GCE and Fe3O4/GCE. The Fe3O4/GCE exhibited a remarkable catalytic and enhancement effect on the reduction of nimesulide. The reduction peak potential of nimesulide shifted positively from –0.683 V at bare GCE to –0.625 V at Fe3O4/GCE, and the sensitivity was increased by ca. 3 times. Some experimental conditions were optimized. The linear range between the peak current and the concentration of nimesulide was 2.6×10–6―1.0×10–4 mol/L(R=0.993) with a detection limit of 1.3×10–7 mol/L. This method has been used to determine the content of nimesulide in medical tablets. The recovery was determined to be 96.9%―101.9% by means of standard addition method. The method is comparable to UV-Vis spectrometry.
    Raman and UV-Vis Spectroscopy Applied to the Analysis of Liver Tissues from Rats with Myocardial Ischemia Induced by Isoproterenol
    GAO Hai-cheng, ZOU Ying-gang, HUANG Yu-xin, GAO Hai-mei, CHEN Lei and PEI Jin*
    2011, 27(4):  570-573. 
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    The application of the laser Raman spectroscopic(LRS) technique for the analysis of liver tissues from rats with myocardial ischemia induced by isoproterenol(ISO) was described. Animal model of myocardial ischemia was established for rats induced by ISO. Rats were randomly divided into four groups as normal group and myocardial ischemia groups. We observed the successful myocardial ischemia model via serum enzymes levels and hematoxylin-eosin(HE) staining, and detected the liver tissue of the rats from normal group and liver tissue of the rats from myocardial ischemia groups via UV-Vis spectroscopy(UV-Vis) and LRS, and the changes of the absorbance spectra were compared in the above four different groups. The results show that ISO can induce rat myocardial ischemia successfully. The spectrum of normal liver tissue supernatant exhibits a strong absorption band at 968 nm,but no absorption band appears in the spectra of liver tissue supernatant solutions from the rats with myocardial ischemia induction after 2, 12 and 72 h presented at 968 nm. LRS results show that Raman intensities of the precipitates suffered from ISO-treatment after 2, 12 and 72 h were obviously increased compared with that of the precipitate of the liver tissue of the normal rats suffered from 0.9 g/L normal saline(NS) treatment. These results indicate that LRS and UV-Vis can be harmless, nondestructive, rapid and effective methods for analyzing different pathological specimens of liver tissue from myocardial ischemia rats.
    Design, Synthesis and Recognition Properties of a New Acetate Ion Receptor Based on Shiff-base Derivative
    QIAO Yan-hong, LIN Hai and LIN Hua-kuan*
    2011, 27(4):  574-577. 
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    A simple colorimetric acetate ion receptor based on 3-methoxysalicylaldehyde-4′-nitrophenyl-hydrazone was synthesized in an ethanol solution through one step, and characterized by 1H NMR and elemental analyses. Its anion-binding ability was examined by UV-Vis absorption spectroscopy in DMSO(dimethyl sulfoxide). The result shows that the compound has a high affinity for and a high selectivity toward acetate ions, and also an advantage of ‘naked-eye’ recognition of color changing from yellow to purple.
    Development of a Derivatization-free GC-FID Method for Evaluation of Free Fatty Acid Levels in Plasma of Diabetic Nephropathy Patients
    HUA Hai-yin, LIANG Qiong-lin*, CHEN Jun, WANG Yi-ming, LI Ping and LUO Guo-an*
    2011, 27(4):  578-583. 
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    Metabolism of free fatty acids(FFAs) is related to several important physiological events and therefore their quantitaion in biological samples arouses extensive interest and efforts. Existing gas chromatography with flame ionization detector(GC-FID) methods for the analysis of FFAs normally require derivatization of them in order to lower boiling points. But this extra procedure tends to induce additional error and it is laborious and time-consuming. A derivatization-free method was therefore established in the present investigation to determine FFAs in human plasma by capillary(GC-FID). After extraction of FFAs from plasma, a highly polar FFAP(free fatty acid in plasma) column was employed to directly quantitate FFAs concentration, free from derivatization reaction. All sample pretreatments were carried out at room temperature, improving recovery of short-chain FFAs. Heptadecanoic acid(C17:0) was employed as internal standard, and the proposed method was validated for recovery, precision, sensitivity, stabi- lity, and linearity. Validation data show that it is suitable for clinical study that has been applied to the evaluation of FFAs levels in plasma of diabetic nephropathy(DN) patients during the course of treatment. Forty-seven patients diagnosed with DN were admitted to the double-blind experiment. Control group(n=17) underwent solely basic treatment and the patients did not show significant change in FFAs concentration during six months of treatment. Experiment group(n=30) was supplied with traditional Chinese medicine besides basic treatment. After six months of medication, their plasma concentration of palmitic acid(C16:0), stearic acid(C18:0) and oleic acid(C18:1n–9) decreased while linolenic acid(C18:3n–3) increased significantly(P<0.05). These four compounds could be served as biomar- kers in the evaluation of drug efficacy, and their quantitation in plasma may provide additional information for   disease progression in DN patients.
    Extraction and Identification of Ginsenoside Re and Its Effects and Mechanism of Protecting Acute Renal Ischemia-reperfusion Injury in Rats
    GU Xin-quan, CHEN Yan-ping, HU Ting-ting, LU Xiu-hua, LI Xi-qian, DU Xiao-hui,..
    2011, 27(4):  584-588. 
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    This paper studies the extraction and identification of ginsenoside Re from ginseng fruits and investigates the effect and mechanism of ginsenoside Re of protecting acute renal ischemia-reperfusion injury in rats. Having been smashed, the ginseng fruits were ultrasonically extracted twice with 95% ethanol for 30 min each. Having been   concentrated, the solution was dissolved with distilled water and separated by two-column chromatography, of  which one was packed with macroporous resin D4020, and the other was packed with macroporous resin D941. The raw product was dissolved with methanol and was purified by elution on a Si gel column, finally ginsenoside Re was        obtained. The structure of the ginsenoside Re was analyzed by the thin-layer chromatography and NMR methods, and HPLC was carried for the content determination. The model of acute renal ischemia-reperfusion injury in rats    was established after ischemia for 1 h and reperfusion for 1 h or 24 h, serum SOD(superoxide dismutase), MDA(malondialdehyde) and plasma TXB2(thromboxane B2) and 6-keto-PGF1α were detected. The results show that it has accurately, fast, convenient merits and so on. Ginsenoside Re has a protective effect on acute renal ischemia-reperfusion injury in rats, the mechanism may be related to improving the imbalance of thromboxane A2(TXA2)/prostacyclin(PGI2) and inhibiting lipid peroxidation reaction.
    Microwave-assisted Palladium-micelle-catalyzed Suzuki Cross-coupling Reaction in Water
    LIN Li, LI Sheng-hai* and JIANG Ri-hua*
    2011, 27(4):  589-591. 
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    A microwave-accelerated Suzuki coupling procedure was developed via guanidinium ionic liquids(GILs) stabilized Pd-micelle. The Pd micelle/GILs play a key role in enhancing the activity, due to the highly dispersed Pd active sites and the phase transfer function of GILs, which ensures the adsorption of reactants and facilitates the translation of the intermediates to the surface of the micelle.
    Biomimetic Epxidation of Alkenes by Phenyliodonium Diacetate [PhI(OAc)2] in the Presence of Mn(III) Complex of meso-Tetrakis(4-methoxyphenylporphyrin) Acetate Mn[(T4-OCH3P)P]OAc
    KARIMIPOUR Gholamreza* and KOWKABI Saeed
    2011, 27(4):  592-594. 
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    We have reported that phenyliodonium diacetate[PhI(OAc)2] can serve as an eco-friendly mild oxidant for the epoxidation of alkenes in the presence of Mn(III) complex of meso-tetrakis(4-methoxyphenylporphyrin) acetate Mn[(T4-OCH3P)P]OAc and imidazole in CH2Cl2. The amounts of the products(%) and selectivities are very dependent upon the electronic and steric properties of the starting alkenes. To evaluate the validity of this catalytic system for C―H activation, cyclohexane and ethylbenzene were oxidized under the epoxidation conditions.
    Syntheses, Crystal Structures and Bioactivities of Two Novel Isatin Derivatives
    SHANG Jian-li, LI Hui-dong, SHANG Jun, SONG Hai-bin, LI Zheng-ming, ...
    2011, 27(4):  595-598. 
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    Two novel compounds 1-(4-fluorobenzyl)-4-chloro-(Z)-3-benzoylhydrazono-2-indolinone(1) and 1-(4-methoxybenzyl)-(Z)-3-benzoylhydrazono-2-indolinone(2) were synthesized and their crystal structures were determined by single-crystal X-ray diffraction. Compound 1(C22H15ClFN3O2) crystallized in the triclinic system, space group P1?? with a=0.94198(19) nm, b=1.4339(3) nm, c=1.5018(3) nm, α=101.58(3)°, β=102.96(3)°, γ=102.73°, V=1.8602(6) nm3, Mr=407.82, Dc=1.456 g/cm3, μ=0.240 mm–1, F(000)=840, Z=4, R1=0.0442 and wR2=0.1064. Compound 2(C23H19N3O3) crystallized in the triclinic system, space group P1?? with a=1.0022(2) nm, b=1.0192(2) nm, c=1.0461(2) nm, α=99.86(3)°, β=117.30(3)°, γ=94.13(3)°, V=0.9215(3) nm3, Mr=385.41, Dc=1.389 g/cm3, μ=0.094 mm–1, F(000)=404, Z=2, R1=0.0403 and wR2=0.1142. The preliminary herbicidal activities of the two compounds were also evaluated.
    Synthesis and Spectroscopic Study of Coumarin Derivatives
    YAO Chun-feng* and ZENG He-ping
    2011, 27(4):  599-603. 
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    Five new coumarin amide derivatives were synthesized with coupling reagent, the structures of which were characterized by IR, 1H NMR, 13C NMR, MS and element analysis. Their spectral properties were studied in dichloromethane,in N,N-dimethylformamide(DMF) and in solid state. Solvent polarity has less influence on the UV-Vis maximum absorption at about 430 nm. The maximum emission wavelengths change from 464 nm to 474 nm in dichloromethane, from 476 nm to 482 nm in DMF and from 521 nm to 548 nm in solid state, respectively. The fluorescence intensities of compounds 8 and 9 were extremely strong in solvents or in solid state. Compounds 8 and 9 exhibited high fluorescenct quantumn yields in solution compared to compounds 4―7. The fluorescence lifetimes of all the compounds in solvents were measured.
    Synthesis of Novel Chiral 7-Amide Substituted-4-androstene-3,17-dione Derivatives
    CHEN Shao-rui, ZHOU Xue-qin, LI Wei and LIU Dong-zhi*
    2011, 27(4):  604-609. 
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    A series of novel 7-amide substituted-4-androstene-3,17-dione derivatives(8αa―8αh or 8βa―8βh) was synthesized from the important intermediates 5 by N-acylation and acidic hydrolysis. Compounds 5α and 5β were obtained through the reaction sequence including acetalization, allylic oxidation, oximation and reduction. The structures of the target compounds were characterized by MS, 1H NMR, 13C NMR and HRMS spectra and their stereo configurations were identified through DEPT(distortionless enhancement by polarization transfer), HMQC(hetero- nuclear multiple quantum coherence) and NOE(nuclear overhauser effect) correlation.
    Synthesis and Insecticidal Activities of Novel Analogues of Chlorantraniliprole Containing Nitro Group
    FENG Qi, WANG Ming-zhong, XIONG Li-xia, LIU Zhi-li and LI Zheng-ming*
    2011, 27(4):  610-613. 
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    Twelve novel analogues of chlorantraniliprole containing nitro group were synthesized, and their structures were characterized by 1H NMR and high-resolution mass spectrometry(HRMS). Their evaluated insecticidal activities against oriental armyworm(Mythimna separata) indicate that the nitro-containing analogues showed favorable insecticidal activities, while the activity of compounds 5g at 0.25 mg/L was 40%, but still lower than chlorantraniliprole.
    Synthesis and Immunosuppressive Activity of New Amino Alcohol Derivatives(I)
    MI Hao-yu, CUI Lin-lin, ZHANG Qun-li, LI Fang, JIANG Tao, LIANG Yong-tao, ...
    2011, 27(4):  614-618. 
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    A series of new amino alcohol derivatives was synthesized and evaluated for their immunosuppressive activity on mouse peripheral blood lymphocytes. The structures were confirmed by means of 1H NMR, 13C NMR, IR and MS. Most of the compounds display moderate to potent inhibitory activity. Compound 9d shows the most activity among them that are expected as a powerful candidate for safer immunosuppressant for organ transplantations and the treatment of autoimmune diseases.
    Selection, Expression and Purification of Human Abzyme Containing Selenium with Type I Thyroxine Deiodinase Activity
    LIN Feng*, LIU Lan-ying and NI Jia-zuan
    2011, 27(4):  619-622. 
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    Single-chain fragment variable(ScFv) antibodies with the substrate binding sites were obtained by repetitive selections from a semi-synthetic phage display antibody library against two haptens——thyroxine(T4) and O-methyl-T4(O-CH3-T4). The positive phage clones were determined by ELISA and then cloned into vector pET30a(+). The recombined plasmids were identified by DNA sequence analysis and transformed into E. coli BL21(DE3). The expressed proteins were isolated, purified, identified by Western blot analysis, and finally the catalytic group——selenocysteine was incorporated into the antibodies binding sites by chemical mutation. The type ? thyroxine deiodinase activities of the abzymes were 0.006 and 0.008 pg·mL–1·min–1, respectively, which were about one tenth that of the mouse intact antibody that was obtained from hybridoma.
    Construction of B7x Gene Overexpression Lentiviral-based Vector
    SUN Mei, JIANG Rui, SUN Li-hua, LI Jin-dong, WANG Bin, JIN Cheng-yan, ...
    2011, 27(4):  623-627. 
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    To construct overexpression lentiviral-based vector carrying rat B7x gene, B7x gene precursor sequences amplified by polymerse chain reaction(PCR) were ligated with pLVTHM to generate pLVTHM-B7x gene expression lentiviral-based vector. The positive clones were selected to be submitted to DNA sequencing. HEK293T cells were co-transfected with pLVTHM-B7x and two packaging plasmids psPAX2 and pMD2G to produce lentivirus which express B7x gene. The mRNA expression levels of B7x gene and virus titer were detected by real-time PCR in HEK293T cells. The B7x protein levels were detected by Western blot analysis in HEK293T cells. The identification of restriction enzyme digestion and DNA sequencing confirmed that pLVTHM-B7x lentiviral-based vector was successfully constructed. Green fluorescence was observed in HEK293T packaging cells by means of an inverted fluorescence microscope and the virus titer measured was 2×108 TU/mL. It will establish the foundation for studing deeply the biological function of B7x to construct successfully B7x expression lentiviral-based vector.
    EPR Effect of Amphiphilic Copolymer Micelles Observed by Fluorescent Imaging
    LIU Tong-jun*, LIU Shi, HU Xiu-li, SHENG Shi-hou, HUANG Yu-bin and JING Xia-bin*
    2011, 27(4):  628-634. 
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    Enhanced permeation and retention(EPR) targeting effect of rhodamine B labeled PEG-b-P(LA-co-DHP) [PEG: poly(ethylene glycol); LA: L-lactide; DHP: 2,2-dihydroxylmethyl-propylene carbonate] micelles(RhB-    micelles) was observed in H22 liver cancer bearing mice. The RhB-micelles were prepared by conjugating rhodamine B with the DHP units of amphiphilic block copolymer PEG-b-P(LA-co-DHP) followed by subsequent self-assembling of the conjugate. The parent copolymer PEG-b-P(LA-co-DHP) was synthesized by ring-opening  copolymerization of LA and DHP with PEG as macroinitiator and diethyl zinc(ZnEt2) as catalyst. The micelles have a spherical shape and the average diameter is ca. 50 nm by TEM(transmission electron microscope) or 80 nm by DLS(dynamic light scattering). Their in vitro cell uptake experiment by CLSM(confocal laser scanning microscopy) and flow cytometry showed preferential internalization of micelles by MCF-7 human breast cancer cells to free RhB. The in vivo tests by live animal imaging and ex vivo excised organ imaging showed that after vena tail injection, free RhB molecules were distributed in the whole body through the circulation system and then gradually metabolized and excreted and there was no preferential partition in tumor bed from the beginning to the end. But the RhB-micelles were preferentially distributed to the tumor bed so that their concentration(fluorescent intensity) in tumor bed got the level of the liver at a certain time point between 1 and 6 h and reached a maximum relative intensity at around 12 h, indicating an obvious EPR effect of RhB-micelles in H22 liver cancer.
    Screening Peptide Inhibitors Using Phage Peptide Library with Isocitrate Lyase in Mycobacterium tuberculosis as Target
    YIN Yu-he, NIU Xue, SUN Bo, TENG Guo-sheng, ZHAO Yun-hui and WU Cong-mei*
    2011, 27(4):  635-640. 
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    When devoured by macrophages, Mycobacterium tuberculosis remains persistent in macrophages and gains energy through the glyoxylate bypass to maintain its long-term existence in host cells. Therefore it is possible to stop persistent infections by interdicting the glyoxylate bypass in which the isocitrate lyase(ICL) is the key rate-limiting enzyme and a persistence factor. ICL is the target of anti-TB(TB: tubercular) drugs, which could screen ICL out and effectively inhibit the activity of ICL in Mycobacterium tuberculosis, and because of this, anti-TB drugs can be used to kill persistent Mycobacterium tuberculosis. In this study, the ICL gene of the Mycobacterium tuberculosis H37Rv was cloned successfully and recombinant protein with bioactivity was obtained through the enzyme characteristic appraisal. The specific activity of the recombined ICL is 24 μmol·mg–1 ·min–1. The recombined ICL protein was used as the target, and phages which can specifically combine to ICL were screened in the phage 7 peptide library. According to the results of the ELISA and DNA sequence detection, eventually three 7-peptide chains were synthesized. Then the peptide chains were reacted with ICL, respectively, to detect their inhibitory effects on ICL. The results show that all the three 7-peptide chains possessed varying inhibitory effects on the activity of ICL. This study provided lead compounds for the research and development of new peptide anti-TB drugs.
    Genetic Diversity Caused by Environmental Stress in Natural Populations of Niupidujuan as Revealed by RAPD Technique
    DU Ying-da, XING Ming, YANG Zhi-yong, LIU Yan-fei and CHEN Xia*
    2011, 27(4):  641-645. 
    Abstract ( )   PDF (183KB) ( )  
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    Multiplex environmental factors are generally expected to have significant effects on genetic diversity of plant populations. In this study, randomly amplified polymorphic DNA(RAPD) technique was used to reveal the genetic diversity in the same species of four populations collected from Niupidujuan(Rhododendron chrysanthum) at different altitudes, an endangered species, endemic to Northeast China. Initially, twenty informative and reproducible primers were chosen for final RAPD analysis. A total of 152 clear bands were obtained, including 143 polymorphic ones. With the help of POPGENE software, the poly rate was calculated to be 94.07% and the evenness of amplified bands for every primer was 6.8. Additionally, the mean observed number of alleles was 1.7265 with an effective number of 1.3608. An examination of the gene indicated a diversity of 0.2162 with an information diversity index of 0.3313. For these data, the clustering blurred analysis was performed with the aid of NTSYS-pc software to define the Nei’s gene diversity and the Shannon information diversity index of the four plant populations. The relationships between the genetic diversity indexes on the one hand and the geographic and climatic factors on the other hand were estimated by the Pearson correlation with SPSS 11.0 software. The results of the correlation analysis show that there were significant(P<0.05) or highly significant(P<0.01) correlations between each of the genetic diversity indexes and the different temperature which were mainly caused by the altitude different populations located. These data highlight the importance of native populations in shaping the spatial genetic structure in Niupidujuan.
    Anti-tumor Effects of a Recombinant Fowlpox Virus Expressing Apoptin on Human Cervical Carcinoma in Vivo and in Vitro
    ZHU Ji-hong, LI Xiao, SUN Li-li, ZHANG Mu-chun, KAN Shi-fu, LIU Lei, ...
    2011, 27(4):  646-650. 
    Abstract ( )   PDF (1025KB) ( )  
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    Apoptin is a chicken anemia virus-derived, p53-independent, bcl-2-insensitive apoptotic protein with the ability to specifically induce apoptosis of various human tumor cells, but not of normal diploid cells. To explore the application of apoptin in tumor gene therapy, we used a recombinant fowlpox virus expressing apoptin protein (vFV-Apoptin) to investigate the anti-tumor effectes of vFV-Apoptin on human cervical carcinoma(HeLa) cells in vivo and in vitro through 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide(MTT) assay, acridine orage/ethidium bromide(AO/EB) and annexin V staining test, respectively. The results show that vFV-Apoptin inhibites the proliferation of HeLa cells in vitro through inducing the apoptosis of HeLa cells, and the inhibition effect of vFV-Apoptin has a dose-effect and time-effect relationship. The results of animal models show that vFV-Apoptin significantly inhibits tumor growth, extends the lifespan of animals and improves the mean survival. Experimental results indicate that vFV-Apoptin has a potential application in the tumor gene therapy.
    Anti-microbial Activities of Protic Ionic Liquids Studied with Microcalorimetry Method
    DING Yang-jun, NIE Yi* and YU You
    2011, 27(4):  651-654. 
    Abstract ( )   PDF (217KB) ( )  
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    The anti-microbial activities of seven protic ionic liquids(ILs) against Escherichia coli and Staphylococcus aureus were studied by a micro-calorimetric method at 310 K. The bacterial growth rate constants were determined based on the bacterial growth power-time curves, and minimum biocidal concentrations were estimated. The results indicate that the protic ILs studied show inhibitory activities on the bacteria, implying a potential eco-toxicity to the microorganisms in the water system. Moreover, the inhibition effect of ionic liquids is related to the structure of the cation and anion of protic ILs. This type of proactive approach could aid in the assessment of the greenness of ILs to be used in the future.
    Discovery of a Novel 5-HT2A Inhibitor by Pharmacophore-based Virtual Screening
    XIONG Zi-jun, DU Peng, LI Bian, XU Li-li, ZHEN Xue-chu* and FU Wei*
    2011, 27(4):  655-660. 
    Abstract ( )   PDF (1035KB) ( )  
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    The serotonin 2A(5-HT2A) receptor has been implicated in several neurological conditions and potent 5-HT2A antagonists have therapeutic effects in the treatment of schizo phrenia and depression. In this study, a potent novel 5-HT2A inhibitor 05245768 with a Ki value of (593.89±34.10) nmol/L was discovered by integrating a set of computational approaches and experiments(protein structure prediction, pharmacophore-based virtual screening, automated molecular docking and pharmacological bioassay). The 5-HT2A receptor showed a negatively charged bin- ding pocket. The binding mode of compound 05245768 with 5-HT2A was obtained by GOLD docking procedure, which revealed the conserved interaction between protonated nitrogen in compound 05245768 and carboxylate group of D3.32 at the active site of 5-HT2A.
    Spectra of Linear Polyene Molecule-canthaxanthin
    OUYANG Shun-li, LI Zuo-wei, CHEN Yuan-zheng, MEN Zhi-wei, WU Nan-nan, ...
    2011, 27(4):  661-663. 
    Abstract ( )   PDF (222KB) ( )  
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    Raman spectra and ultraviolet-visible(UV-Vis) absorption spectra of linear polyene molecule-canthaxan- thin in n-hexane are measured and analyzed. In addition, the optimized structure of canthaxanthin was calculated via density functional theory(DFT) functional B3LYP. With decreasing the concentration, Raman scattering cross section (RSCS) of fundamental frequency is extremely high, and the UV-Vis absorption bands become narrower. The results of coherent weakly damped electron-Lattice vibration model were analyzed.
    Solvent-free Synthesis of Flavanone over New Hybrid Mesoporous Base Catalysts
    WANG Hui, NIU Xiao-di, ZHAO Min, XIAO Xue-bin, WANG Hong-su, ...
    2011, 27(4):  664-668. 
    Abstract ( )   PDF (362KB) ( )  
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    Benzyl and anthracenemethyl groups were respectively bonded to the N atoms of 3-aminopropyl functionalized mesoporous SBA-15(APS-SBA-15) to obtain two new base catalysts over which the condensation reaction of benzaldehyde and 2'-hydroxyacetophenone was studied. Good catalytic activities and high selectivities for flavanones were obtained in solvent-free reactions, which is attributed to the effect of benzyl and anthracenemetyl groups on the base sites of catalysts and the steric hindrance of futher reaction of flavanone with benzaldehyde.
    Structure of Electrochemically Lithiated Li0.33MnO2
    CHEN Hong, WEI Ying-jin, WANG Chun-zhong* and CHEN Gang
    2011, 27(4):  669-672. 
    Abstract ( )   PDF (297KB) ( )  
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    Li0.33MnO2 cathode material was synthesized by solid state reaction which showed a monoclinic structure. In situ synchrotron diffraction showed a reversible structural transition of Li0.33MnO2 during Li+ insertion/extraction. However, it is difficult to determine the crystal structure of lithiated Li0.33+xMnO2 based on their poorly resolved  diffraction patterns. This difficulty was successfully resolved by X-ray absorption spectroscopy(XAS) and Raman scattering which are sensitive to the short-range ordering of material lattice. Extended X-ray absorption fine structure shows the co-existence of corner- and edge-shared [MnO6] in Li0.33MnO2, whereas only edge-shared [MnO6] in   lithiated Li0.96MnO2. Significantly, the differences between the Raman spectra of Li0.33MnO2 and Li0.96MnO2 indicate the phase transition of the material with Li+ intercalation. This work shows that Li0.33MnO2 transforms to monoclinic LiMnO2 with Li+ intercalation. This phase transformation was suggested to evolve from the migration of Mn ions.
    Stereoselectivities in α- and β-Amino Acids Catalyzed Mannich Reactions Involving Cyclohexanone
    WANG Ying, FU Ai-ping*, LI Hong-liang, TIAN Feng-hui, YUAN Shu-ping, ...
    2011, 27(4):  673-677. 
    Abstract ( )   PDF (476KB) ( )  
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    The effects of two different amino acid catalysts on the stereoselectivities in the direct Mannich reactions of cyclohexanone, p-anisidine and p-nitrobenzaldehyde were studied with the aid of density functional theory. Transition states of the stereo-determining C―C bond-forming step with the addition of enamine intermediate to the imine for the L-proline(α-amino acid) and (R)-3-pyrrolidinecarboxylic acid(β-amino acid)-catalyzed processes were reported. B3LYP/6-31G** calculations provide a good explanation for the opposite syn vs. anti diastereoselectivities of these two different kinds of catalysts(syn-selectivity for the α-amino acid catalysts, anti-selectivity for the β-amino acid catalysts). Calculated and observed diastereomeric ratio and enantiomeric excess values are in reasonable agreement.
    Vapor-liquid Phase Equilibria for CO2+Tertpentanol Binary System at Elevated Pressures
    WANG Lin, LUO Jian-cheng, YANG Hao and CHEN Kai-xun*
    2011, 27(4):  678-682. 
    Abstract ( )   PDF (294KB) ( )  
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    Vapor-liquid phase equilibrium data of tertpentanol in carbon dioxide were measured at temperatures of 313.4, 323.4, 333.5 and 343.5 K and in the pressure range of 4.56―11.44 MPa. The phase equilibium apparatus used in the work was a variable-volume high-pressure cell. The experimental data were reasonably correlated with Peng-Robinson equation of state(PR-EOS) together with van der Waals-2 two-parameter mixing rules. Henry’s Law constants and partial molar volumes of CO2 at infinite dilution were estimated with Krichevsky-Kasarnovsky equation, and Henry’s Law constants increase with increasing temperature, however, partial molar volumes of CO2 at infinite dilution are negative whose magnitudes decrease with temperature. Partial molar volumes of CO2 and tertpentanol in liquid phase at equilibrium were calculated.
    Controllable Silver Plating on Silica for Surface-enhanced Raman Scattering
    CHEN Lei, HAN Xiao-xia, YANG Jing-xiu, ZHOU Ji, LU Zhi-cheng, SONG Wei, ...
    2011, 27(4):  683-687. 
    Abstract ( )   PDF (596KB) ( )  
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    We proposed a facile and rapid method for preparing silica-silver core-shell(SSCS) substrates to use Ag electroless plating on SiO2@Au-seed particles. UV-Vis-NIR absorption spectrometer and SEM were employed to monitor the reaction process of the formation of Ag on the surfaces of silica beads, and the optical resonance of the substrate could shift from visible to NIR region. It has been found that surface-enhanced Raman scattering(SERS) enhancement changes with the electroless plating time and the SSCS substrate with the plating time of 90 s(90SSCS) shows the strongest SERS response under the laser excitation at 514.5 nm. Signals collected over multiple spots and substrate of rhodamine 6G(R6G) resulted in a relative standard deviation(RSD) of 9.75%. The calculated enhancement factor(EF) was approximately 105―106. SSCS substrate exhibits high SERS performance, which is due to electromagnetic SERS enhancement with additional localization field within closely packed Ag nanoparticles decorated on the SiO2 nanoparticles. And this substrate presents tunable and broad localized surface plasmon resonance(LSPR), so this method may open a new way for SERS studies with other laser excitation.
    Study of Imidazolium Ionic Liquids: Temperature-dependent Fluorescence and Molecular Dynamics Simulation
    FU Hai-ying, ZHU Guang-lai, WU Guo-zhong*, SHA Mao-lin and DOU Qiang
    2011, 27(4):  688-692. 
    Abstract ( )   PDF (740KB) ( )  
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    The steady-state fluorescence spectra and molecular dynamics simulations were explored to investigate the temperature dependent organization in some imidazolium ionic liquids: 1-butyl-3-methylimidazolium hexafluorophosphate([bmim][PF6]), 1-ethyl-3-methylimidazolium ethylsulfate([emim][EtSO4]) and 1-butyl-3-methylimida- zolium tetrafluoroborate([bmim][BF4]). The pure room temperature ionic liquids(ILs) exhibit a large red shift at more than an excitation wavelength of around 340 nm, which demonstrates the heterogeneous nature of the liquids. Furthermore, the fluorescence spectra of the ionic liquids were found to be temperature-dependent. The emission intensity gradually decreased with increasing temperature for the neat ionic liquids and the mixed solutions of [bmim][BF4]-H2O, which was the special phenomena induced by not only the local structure but also the viscosity. The molecular dynamics simulation further confirms that the structures of ionic liquids are sensitive to the surroun- ding environment because of the aggregation degree of ILs.
    Using Raman Spectroscopy and ab initio Calculations to Investigate Intermolecular Hydrogen Bonds in Binary Mixture(Tetrahydrofuran+Water)
    WU Nan-nan, OUYANG Shun-li, LI Zuo-wei, LIU Jing-yao* and GAO Shu-qin
    2011, 27(4):  693-696. 
    Abstract ( )   PDF (467KB) ( )  
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    We analyzed the properties and structures of the hydrogen-bonded complexes of tetrahydrofuran(THF) and water by means of experimental Raman spectra and ab initio calculations. The optimized geometries and vibrational frequencies of the neat THF molecule and its hydrogen-bonded complexes with water(THF/H2O) were calculated at the MP2/6-311+G(d,p) level of theory. We found that the intermolecular hydrogen bonds which are formed from the binary mixtures of the neat THF and water with different molar ratios could explain the changes in wavenumber position and linewidth very well. The combination of ab initio calculations and experimental Raman spectral data provides an insight into the hydrogen bonds leading to the concentration dependent changes in the spectral features.
    Investigation on Semi-conductive Properties of Hot Growth Film on 316L Austenitic Stainless Steel
    LI Dang-guo, CHEN Da-rong*, WANG Jia-dao and CHEN Hao-sheng
    2011, 27(4):  697-702. 
    Abstract ( )   PDF (806KB) ( )  
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    The semi-conductive performances of hot growth film on 316L stainless steel were studied by means of electrochemical impedance spectroscopy(EIS) and Mott-Schottky analysis. The chemical compositions of the hot growth films were detected by X-ray photoelectron spectroscopy(XPS). The results show that the transfer resistance and film resistance increase with increasing temperature to 400 °C, then they decrease sharply with further continuously increasing temperature. Formation time plays an important role in determining the electron properties of the hot growth film, when formation time reaches to 1 d, the transfer resistance and film resistance reach to the maximum value. Hot growth film has better anticorrosion behaviour in bicarbonate/carbonate solution containing chloride ions or sulfide ions. Mott-Schottky analysis reveals that hot growth film has n-p type semi-conductive property. XPS results show that the hot growth film is mainly composed of the inner chromium oxide and the outer iron oxide.
    Determination of Ni2+ in Waters with Sodium Polyacrylate as a Binding Phase in Diffusive Gradients in Thin-films
    CHEN Hong, DONG Jia, NIU Yong-xin and SUN Ting*
    2011, 27(4):  703-707. 
    Abstract ( )   PDF (195KB) ( )  
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    An aqueous solution containing sodium polyacrylate(PAAS) was used in diffusive gradient in thin-films technique(DGT) to measure DGT-labile Ni2+ concentrations. The DGT devices(PAAS DGT) were validated in four types of solutions, including synthetic river water containing metal ions with complexing EDTA or that without complexing EDTA, natural river water(Ling River, Jinzhou, China) spiked with Ni2+, and an industrial wastewater  (Jinzhou, China). Results show that only free metal ions were measured by PAAS DGT, recovery=97.36% in the solutions containing only free metal ions, recovery=49.62% in a solution with metal/EDTA molar ratio of 2:1 and recovery=0 in the solutions with metal/EDTA molar ratios of 1:1 and 1:2. These indicated that the complexes of Ni-EDTA were DGT-inert. The DGT performance in spiked river water(recovery=18.24%) and in industrial wastewater(recovery=12.25%) were investigated, which indicated that the measurement of metals by this DGT device did not include the humic substances complexed fractions of metals. The binding properties of PAAS DGT for Ni2+ were investigated under different conditions of pH value and ionic strength. Conditional stability constants(lgK) of PAAS-Ni complexes were also evaluated.
    Comparison of BSA Release Behavior from Electrospun PLGA and PLGA/Chitosan Membranes
    WU Li-li, LI Xiao-ran, LI Hua and YUAN Xiao-yan*
    2011, 27(4):  708-711. 
    Abstract ( )   PDF (428KB) ( )  
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    In order to encapsulate and controlled-release bioactive proteins, three fibrous membranes, i.e.,      poly(L-lactide-co-glycolide)(PLGA), hybrid PLGA and chitosan(H-PLGA/CS), and core/shell PLGA/CS (C-PLGA/CS), were produced by emulsion electrospinning, co-electrospinning and coaxial electrospinning, respectively. Bovine serum albumin(BSA) was selected as a model protein. The loading efficiency of BSA in the PLGA membrane was 1.56%, lower than those of H-PLGA/CS(5.98%) and C-PLGA/CS(4.80%). BSA release profiles from the three membranes showed initial burst releases in the first 7 d and then sustained release for 28 d. Cumulative releases at the end of the releasing period, 28 d, from the above three membranes were nearly 63%, 88% and 94%, respectively, indicating that the introduction of chitosan and the core/shell fiber structure could enhance BSA release rate. In addition, all these electrospun membranes could retain their fibrous morphologies after in vitro release of BSA for 28 d.
    Research Note
    Direct Synthesis of Nitriles from Aldehydes in Ionic Liquids
    ZHANG Jun-li, CHEN Xiao-rong, YU Yan-peng, XU Kun-ling and HUANG Jun*
    2011, 27(4):  712-714. 
    Abstract ( )   PDF (211KB) ( )  
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    One-pot synthesis of nitriles from aldehydes with hydroxylamine hydrochloride was developed using [bmim][OTf] as the ionic liquid. The yield of benzonitrile was achieved to 97% at the temperature of 120 oC. [bmim][OTf] can be recovered and reused for this reaction. This method is simple and highly efficient, which makes it an attractive alternative.
Editor-in-Chief:
Jihong YU
ISSN 1005-9040
CN 22-1183/O6
Special Issue/Column
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