Separation, purification and antioxidant activity of polysaccharide from Coprinus comatus / 生物工程学报

We compared the ways of deproteinization for crude polysaccharides of Coprinus comatus, and finally selected Sevage method as the optimal method. Two main fractions of Ccp-I-A and Ccp-I-B were obtained after DEAE-52 cellulose and Sephadex G-200 chromatography, both were white-floc, soluble in water, insoluble in absolute ethyl alcohol, acetone and other organic solvents. Additionally, Fehling reagent, CTAB, Sulphuric acid-carbazole, I-KI and FeCl₃ reaction were all negative. GC analysis showed Ccp-I-A was composed of mannitose, glucose and galactose in molar ratios of 2.03:9.52:1, whereas Ccp-I-B was composed of fucose and galactose with molar ratios of 1:5.21. Antioxidant activity test showed that Ccp-I-A and Ccp-I-B had good scavenging abilities on DPPH and ·OH. Compared to Ccp-I-B,the scavenging activity of Ccp-I-A was much stronger, and the scavenging rate could reach 72.1% and 55.3% respectively when the concentration was 300 μg/mL.

Preparation of nanoparticles of ZL-004 and preliminary study on its pharmacokinetics / 中国药学杂志

OBJECTIVE: To prepare ZL-004 loaded PLGA nanoparticles (ZL-004-NP) and evaluate its release characteristics in vitro and pharmacokinetics in rats. METHODS: In order to determine physico-chemical property of ZL-004, saturation-constant temperature method, potentiometric method and shake flask method were employed to obtain relative parameters, respectively. After investigating single factors, the orthogonal design was used to gain optimal formulation. Then characteristic of ZL-004-NP prepared under condition of best formulation was determined by scanning electronmicroscopy, laser particle size analyzer, dialysis method, respectively. Afterward, amounts of ZL-004 in plasma were determined under UPLC-MS/MS and relative bioavailability between ZL-004-NP and its raw material was calculated. RESULTS: The feature of ZL-004-NP met aim of experiment, which contained smooth spheres, 121.34 nm of diameter, 0.16 of PDI, 89.63% of encapsulation efficiency, 7.65% of loading drug content, sustained release about 216 h in vitro and less burst in initiate stage. In vivo, cmax of ZL-004-NP was increased robustly 1.7 times, which reached to 125 μg · L-1 and tmax was cut down rapidly from (6.47 ± 0.51) h to (4.13 ± 0.48) h, compared with crude ZL-004. Relative bioavailability between ZL-004-NP and crude ZL-004 was 200.99%, which greatly improved effect of oral absorption. CONCLUSION: ZL-004-NP might be developed to improve water-insolube nature of ZL-004, which could increase oral relative bioavailability.

Structural Bioinformatics Analysis of Disease-related Mutations

In order to understand the protein functions that are related to disease, it is important to detect the correlation between amino acid mutations and isease. Many mutation studies about disease-related proteins have been carried out through molecular biology techniques, such as vector design, protein engineering, and protein crystallization. However, experimental protein mutation studies are time-consuming, be it in vivo or in vitro. We therefore performed a bioinformatic analysis of known disease-related mutations and their protein structure changes in order to analyze the correlation between mutation and disease. For this study, we selected 111 diseases that were related to 175 proteins from the PDB database and 710 mutations that were found in the protein structures. The mutations were acquired from the Human Gene Mutation Database (HGMD). We selected point mutations, excluding only insertions or deletions, for detecting structural changes. To detect a structural change by mutation, we analyzed not only the structural properties (distance of pocket and mutation, pocket size, surface size, and stability), but also the physico-chemical properties (weight, instability, isoelectric point (IEP), and GRAVY score) for the 710 mutations. We detected that the distance between the pocket and disease-related mutation lay within 20 A (98.5%, 700 proteins). We found that there was no significant correlation between structural stability and disease-causing mutations or between hydrophobicity changes and critical mutations. For large-scale mutational analysis of disease-causing mutations, our bioinformatics approach, using 710 structural mutations, called "Structural Mutatomics," can help researchers to detect disease-specific mutations and to understand the biological functions of disease-related proteins.

Preparation and Physico-chemical Property of Coenzyme Q_(10) Submicroemulsion / 中国药房

OBJECTIVE:To prepare coenzyme Q10 submicroemulsion and investigate its stability and physico-chemical properties.METHODS:Orthogonal experiment was designed to optimize the formulation and preparation procedure of coenzyme Q10 submicroemulsion.The content and entrapment efficiency of the preparation were determined by HPLC,and its properties such as particle size,? potential,pH value and stability were studied.RESULTS:The optimal formulation and preparation procedure of coenzyme Q10 submicroemulsion were as follows:the ratio of soybean oil to medium-chain triglyceride was 1∶2;the ratio of soybean phospholipids to poloxamer 188 was 3∶1;the high speed shearing emulsification time was 10min and the preparation temperature was 60℃.The mean entrapment efficiency of 3 batches of coenzyme Q10 submicroemulsions was 98.07%,with a ? potential of —28.4mV and a mean particle size of 168 nm.Illumination and freeze thawing should be avoided for the preparation in storing,which showed a satisfactory stability at 4℃.CONCLUSION:The prepared coenzyme Q10 submicroemulsion was up to the standards of intravenous injection preparations.