Advanced Materials Research Vols. 781-784

Abstract: Cryptotanshinone, tanshinone I, and tanshinone IIA were analyzed by High-performance liquid chromatography /electrospray ionization-mass spectrometry. MSⁿ spectra were obtained and optimized by energy collision induced dissociation (CID) from [M+H]⁺ ions, the effect of collision energy on production of fragmental ions were studied and optimal signals were achieved. With the collision energy of 35%, tanshinones have optimal signals of fragmental ions and maximal amounts of product fragment ions respectively. The fragmentation pathways for the compounds were studied, and this information would be helpful for the quantitative and pharmacokinetic analysis of tanshinones.

Abstract: Soil was collected from Huguangyan of Zhanjiang city and regarded as separate material, It was screened thermophilic protease-producing bacteria though dilution flat, transparent circle and Folin method. Strain 0701 isolated was identificated by its morphological features, physiological and biochemical characteristics and 16s rRNA. This experiment also researched on the genetic stability of enzyme producing, and different temperature, pH influence on proteases activity. 25 high temperature-tolerance protease producing strains were isolated, which accounted for 28.1 % of all the isolated strains. Among them, strain 0701 proteases activity was 247 U/mL and it was far higher than that of proteases for other strains and the most optimum reaction temperature and pH of the proteases were 70oC and pH 7.0, respectively, and retained high enzyme activity within 50-80 oC, pH6.5-9.0. The strain 0703 still had the ability to produce protease after consecutive 4 generation, and was identified as Bacillus subtilis by morphological, physiological-biochemical characteristics and 16s rRNA.

Abstract: Microcrystalline Cellulose and Herba Sarcandrae were used to study the properties and relationship of powders before and after blending. The theoretical values of the physical quantities of the binary mixture were calculated based on the random mixing theory, The measured values of particle size specific surface area, pore volume and angle of repose of binary mixture had larger values than that of theoretical calculations. Whereas, tapped density and bulk density had a smaller value. The results suggested that the mixing behavior was not a complete random, arbitrary or simple superposition of the operation for coarse and sticky particles. The mixing process would create novel particles and physical quantities of novel particles exist regular changes.

Abstract: In this study, Tanshinone IIA lipid microspere (Tan-IIA-LM) was successfully prepared by an ultrasonic and high-pressure homogenization method. The particle size, zeta potential, drug entrapment efficiency and morphological properties of Tan-IIA-LM were characterized in detail. A pharmacokinetic comparative study of Tan-IIA-LM with Sodium Tanshinone IIA Silate (STS) injection was developed. A sensitive and specific HPLC method was established to determine the mass concentration of Tan-IIA and STS in rats plasma samples which were compared between Tan-IIA-LM and STS injection after single dose intravenous administration of 18.35μmol/kg and the results were fitted and analyzed by dedicated software. After intravenous administration, the AUC_0- and C_max values of Tan-IIA were 2.14, 2.22 fold higher than STS. The clearance, apparent volume of distribution and mean residence time (MRT) of Tan-IIA were lower (P<0.01) than STS; other pharmacokinetic parameters had no significant deviation (P>0.05). The pharmacokinetic of Tan-IIA, STS have significant differences at the same molar doses and the AUC and C_max values of Tan-IIA were higher compared with STS.

Abstract: Flavonoids are important bioactive natural compounds, so the differentiation and structural characterization of flavonoids are important research topics. This research results provided valuable mass spectral data and reliable information for the identification of different C- and O-glycosylation position flavonoids and their aglycone. The high resolution electrospray ionization quadrupole time-of-flight tandem mass spectrometry was employed to identify one 8-C-glycosyl flavonoid, one 4-O-glycosyl, one 7-O-glycosyl, and two 3-O-glycosyl flavonoids and four aglycone in negative and positive ion mode with collision-induced dissociation. The characteristic ions of different C- and O-glycosylation position flavonoids and their aglycone were summarized and the fragmentation pathways were proposed in negative and positive ion mode. The MS/MS spectra obtained by UHPLC/Q-TOF-MS/MS analysis could be used for the structural characterization and differentiation of these compounds. These results have practical applications for the rapid identification and structural characterization of these compounds present in crude bioactive extracts or mixtures.

Abstract: The chemical composition of the hydrodistilled essential oils from the leaves, fruits and stems of Lantana camara L. from the South China were analyzed by gas chromatography/mass spectrometry. The GC/MS analysis revealed the sesquiterpenoids germacrene D (20.0%), trans-caryophyllene (14.8%) and α-humulene (5.8%) as major constituents in the leaf oil, and palmitic acid (22.5%), stearic acid (12.6%) and germacrene D (7.0%) as major constituents in the fruit oil, while the major constituents in the stem oil were germacrene D (31.0%), β-elemene (17.6%) and α-phellendrene (6.7%). The essential oils were tested for antibacterial activity against 6 strains, using broth dilution method. Both the leaf and stem oils showed moderate inhibitory activity against Staphylococcus aureus, S. epidermidis, Bacillus subtilis, Bacillus dysenteriae, B. proteus, and Escherichia coli. High percentage of sesquiterpenes in the oils were primary responsible for the antibacterial activities.

Abstract: Feizhu powder, which is a classical Chinese medicine prescription collected in the Chinese Veterinary Pharmacopoeia, comprises four kinds of Chinese herbs: Rhizoma dryopteridis crassirhizomatis, Radix polygoni multiflori (prepared), Fructus hordei germinatus and Soybean (fry). According to dose proportion in the prescription and preparation method collected in the Chinese Veterinary Pharmacopoeia, we have prepared the Feizhu powder. In order to research the antibacterial activity in vitro of homemade Feizhu powder, it was extracted with 95% ethanol and water, respectively. Then minimum inhibitory concentration (MIC) of the extract was detected against Streptococcus suis, Staphylococcus aureus and Escherichia coli by micro porous-plate method. The results showed that MICs of Feizhu powder water extract ranged from 250 mg/mL to 500 mg/mL. While MICs of Feizhu powder alcohol extract ranged from 31.25 mg/mL to 62.5 mg/mL. The test data indicated that antimicrobial activity of Feizhu powder alcohol extract is superior to that of water extract, this lay the foundation for the extract of the antibacterial active ingredients and further research on Feizhu powder.

Abstract: The purpose of this research is to isolate sasanquasaponin and its hydrolyzed products from the defatted seeds of C. oleifera and evaluate their antioxidative effects in vitro and in vivo. Their structures were analyzed by UV, MS, ¹H NMR and ¹³C NMR spectra. Free radical scavenging activities in vitro were determined by DPPH and ABTS; antioxidative activities in vivo were assessed by MDA, SOD and GSH-Px levels in serum and brain tissue of mice after intragastric administration of the products at 50, 200 mg/kg/d for 30 days. Acid hydrolyzed product and sapogenin had stronger antioxidative activities than the sasanquasaponin. They eliminated DPPH and ABTS radicals, significantly (p<0.01) improved SOD and GSH-Px activity in blood, and decreased MDA content in brain, but sasanquasaponin and alkaline hydrolyzed product had no significant change (p>0.05). On the basis of molecular structure in sasanquasaponin, hydroxyl and tigloyl groups are helpful to free radical scavenging activity; smaller molecules and hydrophobic property are beneficial to their absorption and antioxidative effects in vivo. Acid hydrolyzed product and sapogenin are more valuable to protect body against damage from free radicals.

Abstract: Two different silver colloids were produced by the reduction of AgNO₃ using sodium citrate (SCAg colloid) or hydroxylamine hydrochloride (HHAg colloid) as reductant. A comparative study was made on SERS spectra (λexc=514.5 nm) of the calf thymus DNA on the two silver colloids in solution of 0.06mol/L NaCl, pH7.0. With the SCAg colloid, the bands at 732, 915, 1340, and 1448 cm^-1 for adenine, and 800, 1180, 1590, 1625 and 1651 cm^-1 for thymidine are dramatically enhanced. It shows that the DNA molecules can be oriented relatively perpendicularly to the SCAg colloidal surface via the N6H₂, N7 of adenine and C2=O of thymine residues. While with the HHAg colloid, most of the in-plane modes of the bases are poorly enhanced, except for some weak bands or shoulders assigned to cytosine and guanine. Thus the DNA molecules is supposed to be adsorbed on the HHAg colloid surface in a tilted mode.

Abstract: This study was undertaken to achieve high expression and preliminary purification of human β-defensin-2 fusion protein to lay a solid foundation for production of human β-defensin-2 using genetic engineering. A prokaryotic expression vector for human β-defensin-2 fusion protein was generated using in vitro gene synthesis before transformation into BL21 (l DE3) plysS TrX-B host bacteria. High expression of TrX-A-HBD-2 fusion protein was induced with IPTG in the bacteria exposed to various expression conditions. The fusion protein then underwent preliminary purification. The protein of interest was released from the genetically engineered bacteria after freezing and thawing. The expression of the target protein accounted for 16.12% of the total bacterial proteins. Fractional precipitation with saturated ammonium sulfate and metal chelate affinity chromatography yielded human β-defensin-2 peptide fusion protein, with a relative purity of 80.53%.Human β-defensin-2 fusion protein could be highly expressed in a soluble form, with a relatively high purity