Search

Aim: To construct a vector of ErbB2 small interfering RNA(siRNA), and to investigate its effect on ErbB2 expression and the growth of ZR75-1 breast cancer cells., Methods: Two ErbB2-siRNAs were designed and inserted into the pSliencer 2.1-U6 neo vector. After confirmed by restriction and DNA sequencing, siRNA vectors were co-transfected with the FLAG-ErbB2 expression vector into human embryonic kidney 293T cells, or transfected into SKBR3 and ZR75-1 breast cancer cells. The effects of siRNAs on the expression of ErbB2 were identified by Western blot and the proliferation of ZR75-1 cells was repressed., Results: Two siRNAs could effectively inhibit the expression of exogenous and endogenous ErbB2 proteins, and could repress the growth of ZR75-1 cells., Conclusion: ErbB2 siRNAs effectively inhibit the expression of ErbB2, thus repressing the growth of ZR75-1 cells.

Aim: To construct human Egr-1 promoter luciferase reporter system and study its activity induced by ionizing radiation., Methods: Egr-1 promoter was obtained by human genomic PCR and cloned into pGL3-basic vector. After transfection of recombinant plasmid into human tumor cells, the Egr-1 promoter activity induced by ionizing radiation was detected by luciferase reporter assay., Results: The luciferase reporter system of Egr-1 promoter was successfully constructed. The activity of Egr-1 promoter was substantially increased after different doses of IR and reached to the peak at the time point of 48 h after IR., Conclusion: The Egr-1 promoter was constructed in this study showed IR inducible activity in tumor cells, laying foundation for the research of radiation. mediated gene therapy.

The mol-ecule of the title binuclear copper(II) complex, [Cu(2)(C(14)H(8)O(4))(2)(C(12)H(12)N(2))(2)(H(2)O)(2)], is bis-ected by a crystallographic twofold axis. Each Cu(II) atom is coordinated in a distorted octa-hedral geometry by three O atoms from two biphen-yl-2,2'-dicarboxyl-ate anions, one aqua O atom and two N atoms of a 4,4'-dimethyl-2,2'-bipyridine ligand. Intramolecular O-H⋯O hydrogen bonds between the coordinated water molecules and the carboxylate O atoms are also present.

Objective: To study the pharmacological properties of Tougu Xiaotong Granule (TGXTG) in preventing and treating knee osteoarthritis (KOA) at the molecular level., Methods: The computational methods, including principal component analysis, molecular docking, target-ligand space distribution, and the predictions of absorption, distribution, metabolism, excretion and toxicity (ADMET), were introduced to characterize the molecules in TGXTG., Results: The structural properties of molecules in TGXTG were more: diverse than those of the drug/drug-like molecules, and TGXTG could interact with significant target enzymes related to KOA. In addition, the cluster of effective components was preliminarily identified by the target-ligand space distributions. As to the results of ADMET properties, some of them were unsatisfactory, and were merely regarded as references here., Conclusion: Based on this computational pharmacology study, TGXTG is a broad-spectrum recipe inhibiting many important target enzymes, which could effectively postpone the degeneration of spectrum cartilage by coordinately inhibiting the biological effects of cytokines, matrix metallopeptidase 3, and oxygen free radicals. radicals.

Calibration transfer is an important issue to building up universal and comparable performance of spectrometer data in near infrared spectral analysis technology. Methods of slope/bias correction, direct standardization (DS), and target factor analysis (TFA) were used for the calibration transfer among five NIR filter spectrophotometers using maize as the samples. The effects of three calibration transfer methods were compared. The DS method has the best performance. The average calibration transfer difference of DS is 7.01%. This study also relates to the dependence of calibration transfer on the number of standardization samples. It was proven by experiment that the results of calibration transfer will be better as the number of samples is increased and will be generally stable when there are twenty standardization samples. However, the effect of calibration transfer attained by DS is degraded sharply when the number of standardization samples decreases to be below twenty. Moreover, slope/bias and TFA are not sensitive to the number of standardization samples.

Correct data without noise is the basis for obtaining useful information and knowledge. The database system of Chinese materia medica is the application of database technology in the field of Chinese materia medica. Its establishment is based on the analysis, processing and supplementary of much irregular raw data during studying Chinese materia medica information. This paper reviewed several key problems in the data processing of Chinese materia medica based on the information system of Chinese herbal medicine that we are constructing.

Objective: To study the chemical constituents from Rabdosia rubescens., Methods: Compounds were isolated by Silica gel,Sephadex LH-20 and ODS column chromatographies, their strctures were identified by physicochemical property and NMR, MS., Results: Eight constituents were identified as phaeophytin a (1), phaeophytin b (2), 17c-ethoxypheaophorbide a (3), 17c-ethoxyphea ophorbide b (4), oleanolic acid (5), physcion (6), emodin-8-O-beta-D-glucopyranoside (7), isorhamnetin (8)., Conclusion: Compounds 1-4, 6 and 7 are obtained for the first time from this plant.

To study the molecule recognition capability of corilagin-molecularly imprinted polymer (MIP) by the high performance liquid chromatography (HPLC), the molecularly imprinted polymer was synthesized by using corilagin as the template. Chromatographic performance of corilagin was investigated in different mobile phases. The MIP was investigated for the recognition of corilagin and its derivatives and other compounds in the same mobile phase. The MIP exhibited very high affinity for corilagin in the mobile phase of acetonitrile. The K' value will be reduced when the content of polar solvent increased in the mobile phase. The MIP has good selectivity in the mobile phase of acetonitrile-methanol (95:5), but it has no affinity for corilagin's derivatives. The corilagin-MIP has good selectivity for corilagin and it can be used in extracting corilagin and its analogs from herbs.

It is very important to evaluate the fresh sample of alfalfa silage using near infrared reflectance spectroscopy technology (NIRS) for animal production. The nutrient content of forage means the contents of dry matter (DM), crude protein (CP), neutral detergent fiber (NDF), and acid detergent fiber (ADF) in the forage. Because of the high moisture content, it is difficult to make uniform samples for fresh forage and to get useful information from the spectrum. Therefore, it is hard to use NIRS analysis. In order to evaluate the feasibility of using NIRS to analyse the fresh alfalfa silage, the DM, CP, NDF and ADF contents of fresh alfalfa silage were evaluated by the near infrared reflectance spectroscopy model in this experiment using partial least square regression (PLS), Fourier transform technology and sample preparation with liquid nitrogen technology. The analysis samples were obtained through different cultivars, maturity, cuttings and ensiling method. The cross validation was determined between 0.884 6-0. 989 8. The standard error of cross validation was between 3.9 and 9.7 g x kg(-1) fresh weight. Fifty samples were used to test the performance of the models. The coefficients of correlation between the chemical value and the NIRS value are between 0.939 7 and 0.994 9, and the root mean square errors of prediction are between 1.9 and 8.3 g x kg(-1) fresh weight. The results showed that NIRS could be used to evaluate the nutrition of fresh forage.

Aim: To prepare specific single-epitope antibody using immunomagnetic beads coated with multiple antigen peptides(MAP)., Methods: Eight-branch MAP is synthesized by Fmoc SPPS. After immunizing the BALB/c mice with the MAP, polyclonal antiserum was obtained. Then, the immunomagnetic beads (IB) were prepared by coating with the MAP in covalent bond form and were used to purify the specific single-epitope antibody from the antiserum of immunized animals., Results: The titer of antiserum was 1:12,800, which showed the strong immunogenicity of MAP. The best coating concentration of MAP was 100 mg/L and the max coating efficiency was 79%. After purifying specific antibody from polyclonal antiserum using the IB, the obtained antibodies only interacted with one epitope rather than other epitopes. The purity of the antibody reached 95% and the mean recovery rate was 5.8%., Conclusion: The beads coated with MAP can be used to quickly isolate specific single-epitope antibody which is similar to monoclonal antibody. This method provide a convenient shortcut in preparing small amount of antibody with high specificity.

The rapid development of computer technology, molecular pharmacology and molecular biology extremely promoted the extensive and successful application of virtual screening technique (VST) in pharmaceutical exploitation. Based on plentiful literature and their own previous work, the authors elaborated the principles, methodology and strategy of VST systematically, gave a retrospection on the application of VST in the field of TCM in the last several years, and supposed that along with deepening of understanding, VST would play a greater role in TCM.

Objective: To investigate the flavonoids of Oxytropisfalcata., Method: Compounds were isolated by column chromatography using silica gel, Sephadex LH -20 and ODS as the adsorbents. Their structures were elucidated by NMR and MS spectroscopic data., Result: Eight compounds were isolated and elucidated as 2', 4'-dihydroxy-4-methoxy chalcone (1), 2', 4'-dihydroxy chalcone (2), 5,7-dihydroxy-4'-methoxy flavonol (3), 7-hydroxy-4'-methoxy flavanones (4), 3', 7-dihydroxy-2',4'-dimethoxy isoflavan (5), 2'-hydroxy-4'-methoxy chalcone (6), 2'-methoxy-4'-hydroxy chalcone (7), 2',4'-dihydroxy dihydrochalcone (8)., Conclusion: All compounds were obtained from O. falcata for the first time.

BRD7 is a potential nuclear transcription regulation factor related to nasopharyngeal carcinoma (NPC). BRD2, a putative BRD7-interacting protein, has been screened from human fetal brain cDNA library by yeast two-hybrid system. This study was to further identify the interaction between BRD7 and BRD2 in mammalian cells, and to investigate the subcellular localization of BRD2, as well as the effect on the functions of cell biology. Both immunoprecipitation and subcellular colocalization were performed together to identify the interaction of BRD7 with full-length BRD2, as well as C-terminal truncated BRD2 or N-terminal truncated BRD2. GFP direct fluorescence and Hochest 33258 staining were used to investigate the cellular localization pattern of BRD2 and the roles in initiating cell apoptosis in COS7 and HNE1. The results showed that BRD7 could interact with BRD2 and the region from amino acid 430 to 798 of BRD2 was critical for the interaction of BRD2 with BRD7. BRD2 mainly localizes in nucleus in two distribution patterns, diffused and dotted, and BRD2 has distinct roles in initiating apoptosis, and the dotted distribution pattern of BRD2 in nucleus may be a morphologic marker of cell apoptosis.

Objective: Determine the content of ursolic acid of Liuwei Dihuangwan., Method: Using NIR with PLS, PCA-BPANN and WT-BPANN., Result: The predication recovery were 100.7%, 100.6%, 100.1%, and the RSD were 5.42%, 6.49%, 6.52% respectively., Conclusion: NIR can be used in the determination of ursolic acid, which set up the foundation of on-line control of traditional Chinese medicine.

NIR spectromneter for non-destruction measurement of oil contents in an integrated kernel of corn was manufactured. Using LED (light emitting diode) as the light source and six filters as the monochromator, the specifications of the instrument are compared with those of the commercial instruments. The regression coefficient, the standard error, and the relative error of measuring oil contents in an integrated kernel of corn are 0.9688, 0.72 and 0.062 respectively. The results meet the demand of high-oil corn breeding.

In this paper, the absorption spectra of free chlorine in the visible region has been measured after free chlorine in the solution had reacted with N,N-diethyl-1,4-phenylenediamine (DPD). The relation between the absorption spectra and peak wavelength has been discussed. From the experiment of liquor with different concentration, the error of spectral measurement is less than 0.04%. By nonlinear data fitting, the relation between the concentration of free chlorine and the transmittivity has been given.

Here we report molecular dynamics (MD) and free energy perturbation (FEP) simulations applied to hydroxamate-matrix metalloproteinase-2 (MMP-2) complex systems. We have developed some new force field parameters for the hydroxamate functional group that were not included in the AMBER94 force field but were necessary in our simulations. For the representation of the active zinc center, a bonded model was adopted in which restrained electrostatic potential fitting (RESP) charges were used as the electrostatic representation of this model. Using the resulted bonded model, FEP simulations predict the relative binding free energy in good agreement with the experimental value. By analyzing the molecular dynamics (MD) trajectories of the two complex systems, we can provide an explanation of why one of the two inhibitors is favored over the other. The results provide a chemical insight into the interactions between inhibitor and enzyme, and can indicate changes in the inhibitor that would enhance inhibitor-enzyme interactions.

Aim: To establish an HPLC-fluorescence method for determination of loratadine in human plasma and evaluate its relative bioavailability., Methods: An Alltech-C18 column and a mobile phase of acetonitrile-water-glacial acetic acid-triethylamine (90:100:6:0.15) were used. The fluorescence detector was set at Ex 274 nm, Em 450 nm. The flow rate was 1 mL.min-1., Results: The calibration curve was linear over a concentration range of 0.2-30 micrograms.L-1. The limit of quantification was 0.2 microgram.L-1. The average method recoveries varied from 96% to 98%. The results showed AUC, Tmax, Cmax and T1/2 beta between the testing tablets, testing capsules and reference tablets had no significant difference (P > 0.05). Relative bioavailabilities were 107% +/- 17% and 100% +/- 14% respectively., Conclusion: The three formulations were bioequivalent.