Aspergillus sp136抗细菌耐药活性成分研究

基于广谱抗细菌耐药性这一思路，本研究中心建立了一套抗细菌耐药性化合物的筛选方法。由此从3000多种西南地区特殊生境的微生物和植物样品提取物中筛选获得17个抗细菌耐药性活性样品。对其中一株来自峨嵋山土样的微生物（Aspergillus sp136）进行了深入研究。通过TLC自显影等方法从其发酵产物中追踪分离得到抗耐药有效成分，并鉴定为烟曲霉酸。 采用多种方法对烟曲霉酸的体外抗细菌耐药活性进行评价。在平板扩散法中，烟曲霉酸表现出对青霉素（β-内酰氨抗生素）的协同抗耐药能力，其活性大约3倍于克拉维酸。在MIC的测试实验中，烟曲霉酸表现出对青霉素（β-内酰氨抗生素）以及非β-内酰氨抗生素如红霉素、四环素、氯霉素、链霉素、卡那霉素、庆大霉素的抗耐药能力。在棋盘格杀菌以及时间致死曲线的研究中，烟曲霉酸也表现出对青霉素、红霉素、四环素的协同抗细菌耐药活性。 在广泛的活性筛选中发现烟曲霉酸对LDLR基因具有上调活性，表明烟曲霉酸可能具有降血脂的活性。 在研究中发现，同空白对照相比，烟曲霉酸使耐药菌（Bacillus cereus NCPF63509）细胞外β-内酰胺酶酶活大幅度下降，而细胞内β-内酰胺酶酶活仅略有上升，这表明烟曲霉酸对β-内酰胺酶分泌过程具有抑制作用。 综述了β-内酰胺酶的研究进展。 A two-step agar diffusion method was established to screen wide spectrum synergistic antibacterial agents. By using this method, 17 active samples against antibiotic resistance were discovered from more than 3000 plants and microbes, which were collected from southwest china. One isolate Aspergillus sp136 collected from E-mei mountain area was selected for further studies. From the metabolites of this strain, a synergistic antibacterial compound was isolated by bioautographic TLC assay-guided fractionation and identified as helvolic acid. The synergistic effect of helvolic acid was confirmed by several methods in vitro. The synergistic effect of helvolic acid with penicillin (β-lactam antibiotics) was about 3 times as that of clavulanic acid with penicillin in agar diffusion assay. In MIC studies, helvolic acid exhibited synergistic effects with β-lactam antibiotics such as penicillin and non β-lactam antibiotics such as erythromycin, tetracycline, kanamycin, streptomycin and gentamycin. In checkerboard and time-kill studies, helvolic acid also exhibited synergistic effects with penicillin, erythromycin and tetracycline. In general screen of bioactivities, helvolic acid upregulate LDLR gene, which was indirectly determined by the activity of fluorescent enzyme. Therefore, helvolic acid might have the ability to lower lipid in blood. Compared with blank control, the extracellular β-lactamase activity decrease significantly and the intracellular β-lactamase activity increase slightly in Bacillus cereus NCPF63509 in the presence of helvolic acid, indicating that the secretion of β-lactamase was inhibited by helvolic acid. The research of β-lactamase was reviewed.

Adenovirus-mediated wild-type P53 Transfer radiosensitizes H1299 cells to subclinical-dose carbon-ion irradiation through the restoration of p53 function

To determine whether adenovirus-mediated wild-type p53 transfer after radiotherapy could radiosensitize non-small-cell lung cancer (NSCLC) cells to subclinical-dose carbon-ion beam (C-beam), H1299 cells were exposed to a C-beam or -ray and then infected with 5 MOI of AdCMV-p53 or GFP (C-beam or -ray with p53 or GFP).Cell cycle was detected by flow cytometric analysis. The apoptosis was examined by a fluorescent microscope with DAPI staining. DNA fragmentation was monitored by the TUNEL assay. P53 mRNA was detected by reverse-transcriptase polymerase chain reaction. The expression of p53, MDM2, and p21 was monitored by Western blot. Survival fractions were determined by colony-forming assay. The percentages of G1-phase cells in C-beam with p53 increased by 8.2%–16.0%, 5.2%–7.0%, and 5.8%–18.9%, respectively, compared with C-beam only, -ray with p53, or p53 only. The accumulation of G2-phase cells in C-beam with p53 increased by 5.7%–8.9% and 8.8%–14.8%, compared with those in -ray with p53 or p53 only, respectively. The percentage of apoptosis for C-beam with p53 increased by 7.4%–19.1%, 5.8%–11.7%, and 5.2%–19.2%, respectively, compared with C-beam only, -ray with p53, or p53 only. The level of p53 mRNA in C-beam with p53 was significantly higher than that in p53 only. The expression level of p53 and p21 in C-beam with p53 was significantly higher than that in both C-beam with GFP and p53 only. The survival fractions for C-beam with p53 were significantly less than those for the other groups (p 0.05). The data suggested that AdCMV-p53 transfer could more efficiently radiosensitize H1299 cells to subclinical-dose C-beam irradiation through the restoration of p53 function.

Commissioning of electron cooling in CSRm

A new generation electron cooler has started operation in the heavy ion synchrotron CSRm which is used to increase the intensity of heavy ions. Transverse cooling of the ion beam after horizontal multi-turn injection allows beam accumulation at the injection energy. After optimization of the accumulation process an intensity increase in a synchrotron pulse by more than one order of magnitude has been achieved. In given accumulation time interval of 10 seconds, 108particles have been accumulated and accelerated to the final energy. The momentum spread after accumulation and acceleration in the 10−4 range has been demonstrated in six species of ion beams. Primary measurements of accumulation process varying with electron energy,electron beam current, electron beam profile, expansion factor and injection interval have been performed.The lifetimes of ion beams in the presence of electron beams were roughly measured with the help of DCCT signal.

Gound State Properties of Nuclei in (295)118 alpha-Decay Chain

The properties of nuclei belonging to the alpha-decay chain of superheavy element (295)118 have been studied in the framework of axially deformed relativistic mean field (RMF) theory with the parameter set of NL-Z2 in the blocked BCS approximation. Some ground state properties such as binding energies, deformations, and alpha-decay energies Q(alpha) have been obtained and agree well with those from finite-range droplet model (FRDM). The single-particle spectra of nuclei in (295)118 alpha-decay chain show that the shell gaps present obviously nucleon number dependence. The root-mean-square (rms) radii of proton, neutron and matter distributions change slowly from (283)112 to (295)118 but dramatically from (279)110 to (283)112, which may be due to the subshell closure at Z = 110 in (279)110. The alpha-decay half-lives in (295)118 decay chain are evaluated by employing the cluster model and the generalized liquid drop model (GLDM), and the overall agreement is found when they are compared with the known experimental data. The alpha-decay lifetimes obtained from the cluster model are slightly larger than those of GLDM ones. Finally, we predict the alpha-decay half-lives of Z = 118, 116, 114, 112 isotopes using the cluster model and GLDM, which also indicate these two models can corroborate each other in studies on superheavy nuclei. The results from GLDM are always lower than those obtained from the cluster model.

Investigation of PL properties of C-doped SiO2/Si samples after high energy Pb ion irradiation

Amorphous SiO2 thin films with about 400-500 nm in thickness were thermally grown on single crystalline silicon. These SiO2/Si samples were firstly implanted at room temperature (RT) with 100 keV carbon ions to 2.0 x 10(17),5.0 X 10(17) or 1.2 x 10(18) ions/cm(2), then irradiated at RT by 853 MeV Pb ions to 5.0 x 10(11), 1.0 X.10(12) 2.0 x 10(12) or 5.0 x 10(12) ions/cm(2), respectively. The variation of photoluminescence (PL) properties of these samples was analyzed at RT using a fluorescent spectroscopy. The obtained results showed that Pb-ion irradiations led to significant changes of the PL properties of the carbon ion implanted SiO2 films. For examples, 5.0 x 10(12) Pb-ions/cm(2) irradiation produced huge blue and green light-emitters in 2.0 x 10(17) C-ions/cm(2) implanted samples, which resulted in the appearance of two intense PL peaks at about 2.64 and 2.19 eV. For 5.0 x 10(17) carbon-ions/cm(2) implanted samples, 2.0 x 10(12) Pb-ions/cm(2) irradiation could induce the formation of a strong and wide violet band at about 2.90 eV, whereas 5.0 x 10(12) Pb-ionS/cm(2) irradiation could,create double peaks of light emissions at about 2.23 and 2.83 eV. There is no observable PL peak in the 1.2 x 10(18) carbon-ions/cm(2) implanted samples whether it was irradiated with Pb ions or not. All these results implied that special light emitters could be achieved by using proper ion implantation and irradiation conditions, and it will be very useful for the synthesis of new type Of SiO2-based light-emission materials.

Structural and photoluminescent properties of ZnO films deposited by radio frequency reactive sputtering

Zinc oxide films with c-axis preferred orientation were deposited on silicon (100) substrates by radio frequency (RF) reactive sputtering. The properties of the samples were characterized by X-ray diffractometer, X-ray photoelectron spectroscopy and fluorescent-spectrophotometer. The effect of sputtering power and substrate temperature on the structural and photoluminescent (PL) properties of the ZnO films was investigated. The results indicated that when the sputtering power is 100 W and the substrate temperature is 300-400 degrees C, it is suitable for the growth of high c-axis orientation and small strain ZnO films. A violet peak at about 380 nm and a blue band at about 430 nm were observed in the room temperature photoluminescence spectra, and the origin of blue emission was investigated.

Alpha decay half-lives of new superheavy elements through quasimolecular shapes

The lifetimes of alpha decays of the recently produced isotopes of the elements 112, 114, 116 and the element (294)118 and of some decay products have been calculated theoretically within the Wentzel-Kramers-Brillouin approximation. The alpha decay barriers have been determined in the quasimolecular shape path within a generalized liquid drop model including the proximity effects between nuclei in a neck, the mass and charge asymmetry and the precise nuclear radius. These calculations provide reasonable estimated for the observed alpha decay lifetimes. The calculated results have been compared with the results of the density-dependent M3Y effective interaction and the experimental data. It is indicated that the theoretical foundation of the generalized liquid drop model is as good as that of the microscopic DDM3Y model, at least in the sense of predicting the T-1/2 values as long as one uses a correct alpha decay energy. The half lives of these new nuclei are well tested from the consistence of the macroscopic, the microscopic and the experimental data.

Theoretical study on properties of new isotope (265)Bh

The properties of nuclei belonging to the newly observed a-decay chain starting from (265)Bh have been studied. The axially deformed relativistic mean-field calculation with the force NL-Z2 has been performed in the blocked BCS approximation. Some ground state properties such as binding energies, deformations, spins, and parities, as well as Q-values of the alpha-decay for this decay chain have been calculated and compared with known experimental data. Good agreement is found. The single-particle spectrum of the nucleus (265)Bh is studied and some new magic numbers are found, while the magnitudes of the shell gaps in superheavy nuclei are much smaller than those of nuclei before the actinium region, and the Fermi surfaces are close to the continuum. Thus the superheavy nuclei are usually not stable. The alpha-decay lifetimes in the (265)Bh decay chain are evaluated by different formulae, and compared with experimental data. The methods which give good agreement with the data are selected.

High-efficiency transfer and expression of AdCMV-p53 in human hepatocellular carcinoma cells induced by low-dose carbon-ion radiation

Objective To investigate whether the irradiation with C-beam could enhance adenovirus-mediated transfer and expression of p53 in human hepatocellular carcinoma. Materials and methods HepG2 cells were exposed to C-beam or gamma-ray and then infected with replicationdeficient adenovirus recombinant vectors containing human wild-type p53 or green fluorescent protein, respectively. The transfer efficiency and expression level of the exogenous gene were detected by flow cytometric analysis. Cell survival fraction was detected by clonogenic assay. Results The transfer frequency in C-beam or gamma-irradiated groups increased by 50-83% and 5.7-38.0% compared with the control, respectively (P < 0.05). Compared with C-beam alone, p53 alone, and gamma-ray with p53, the percentages of p53 positive cells for 1 Gy C-beam with p53 increased by 56.0-72.0%, 63.5-82.0%, and 31.3-72.5% on first and third day after the treatments, respectively (P < 0.05). The survival fractions for the 2Gy C-bearn and AdCMV-p53 infection groups decreased to similar to 2%. Conclusion C-beam irradiation could significantly promote AdCMV-green fluorescent protein transfer and expression of p53.

Dipole bands in nucleus Pm-139

High-spin states in nucleus Pm-139 have been studied using the reaction Cd-116(Al-27, 4n)Pm-139. Two dipole cascades have been found. Spin and parity assignments were based on the Directional Correlation of Oriented Nuclei (DCO) ratios and systematic behavior in neighboring odd-proton nuclei. The level structures of Pm-139 are compared with those of the N = 78 isotone Eu-141 in which two dipole bands have been confirmed as magnetic rotational bands. The close similarity between them suggests that the dipole bands in Pm-139 may be magnetic rotational bands.

Relativistic Mean Field Study of the Z=117 Isotopic Chain

The properties of the Z = 117 isotopic chain are studied within the framework of the axially deformed relativistic mean field theory (RMFT) in the blocked BCS approximation. The ground-state properties, such as binging energies, deformations as well as the possible.. decay energies and lifetimes are calculated with the parameter set of NL-Z2 and compared with results from the finite range droplet model. The analysis by RMFT shows that the isotopes in the range of mass number A = 291 similar to 300 exhibit higher stability, which suggests that they may be promising nuclei to be hopefully synthesized in the lab among the nuclei Z = 117.

Retention modeling and simultaneous optimization of pH value and gradient steepness in RP-HPLC using feed-forward neural networks

A novel approach is proposed for the simultaneous optimization of mobile phase pH and gradient steepness in RP-HPLC using artificial neural networks. By presetting the initial and final concentration of the organic solvent, a limited number of experiments with different gradient time and pH value of mobile phase are arranged in the two-dimensional space of mobile phase parameters. The retention behavior of each solute is modeled using an individual artificial neural network. An "early stopping" strategy is adopted to ensure the predicting capability of neural networks. The trained neural networks can be used to predict the retention time of solutes under arbitrary mobile phase conditions in the optimization region. Finally, the optimal separation conditions can be found according to a global resolution function. The effectiveness of this method is validated by optimization of separation conditions for amino acids derivatised by a new fluorescent reagent.

Simultaneous genotyping of multiplex single nucleotide polymorphisms of the K-ras gene with a home-made kit

Accurate and fast genotyping of single nucleotide polymorphisms (SNPs) is important in the human genome project. Here an automated fluorescent method that can rapidly and accurately genotype multiplex known SNPs was developed by using a homemade kit, which has lower cost but higher resolution than commercial kit. With this method, oncogene K-ras was investigated, four known SNPs of K-ras gene exon 1 in 31 coloerctal cancer patients were detected. Results indicate that mutations were present in 8(26%) of 31 patients, and most mutations were localized in codon 12. The presence of these mutations is thought to be a critical step and plays an important role in human colorectal carcinogenesisas. (C) 2003 Elsevier B.V. All rights reserved.

Highly efficient separation of dsDNA fragments on glass chips by using an ultralow viscosity sieving matrix

A novel protocol has been established to separate dsDNA fragments with high efficiency on glass chips by using an ultralow viscosity sieving matrix with added glucose. Low-molecular-weight hydroxypropylmethylcellulose (HPMC), with a viscosity nearly equivalent to that of water, was used to electrophoretically separate fluorescent inter-calator-labeled double-stranded DNA (dsDNA) fragments on microfluidic glass chips. In comparison with conventional sieving protocols, low-molecular-weight HPMC as sieving matrix could result in reduced running cost and analysis time, in addition to a comparable separation efficiency of dsDNA fragments. In this paper, the addition of glucose was investigated to enhance the separation of DNA in the lowest viscosity polymer evaluated. The effect of staining dye and field strength were also evaluated. At an applied electric field strength of 200 V/cm, satisfactory resolution of the PBR322/HaeIII DNA marker could be achieved within 4 min by using 2% HPMC-5 with 6% glucose added. Coelectrophoresing PCR product along with phiX174/HaeIII DNA sizing marker was also demonstrated by using the ultralow viscosity HPMC-5 solution on a glass chip.

Analysis of aromatic amines by high-performance liquid chromatography with pre-column derivatization by 2-(9-carbazole)-ethyl-chloroformate

2-(9-Carbazole)-ethyl-chloroformate (CEOC), a novel pre-column fluorescence derivatization reagent, has been developed for the analysis of aromatic amines. Taking five monocyclic aromatic amines (o-toluidine, aniline, 3,4-dimethylaniline, N-ethyl-p-toluidine, and p-phenylenediamine) as testing compounds, derivatization conditions such as pH of borate buffer, reaction time and fluorescent tagging reagent concentration have been investigated. By a one-step procedure, CEOC reacts readily with the aromatic amines to form stable derivatives with excitation and emission wavelengths, respectively, at 293 and 360 nm. This derivatization reaction could be finished within 20 min even at room temperature. The peak shapes of the derivatized aromatic amines can be improved greatly without any addition of competition amines into the mobile phase. Furthermore, this method can offer excellent quantitative precision with high tolerance of the matrix of samples. (C) 2003 Elsevier B.V. All rights reserved.