Construction of recombinant adenovirus expressing capsid protein of serotype O foot-and-mouth disease virus and analysis of its immunogenicity / 生物工程学报

In order to construct a recombinant replication deficient human type 5 adenovirus (Ad5) expressing a foot-and-mouth disease virus (FMDV) capsid protein, specific primers for P12A and 3B3C genes of FMDV-OZK93 were synthesized. The P12A and 3B3C genes were then amplified and connected by fusion PCR, and a recombinant shuttle plasmid pDC316-mCMV-EGFP-P12A3B3C expressing the FMDV-OZK93 capsid protein precursor P12A and 3B3C protease were obtained by inserting the P12A3B3C gene into the pDC316-mCMV-EGFP plasmid. The recombinant adenovirus rAdv-P12A3B3C-OZK93 was subsequently packaged, characterized and amplified using AdMaxTM adenovirus packaging system, and the expression was verified by infecting human embryonic kidney cell HEK-293. The humoral and cellular immunity levels of well-expressed and purified recombinant adenovirus immunized mice were evaluated. The results showed that rAdv-P12A3B3C-OZK93 could be stably passaged and the maximum virus titer reached 1×109.1 TCID50/mL. Western blotting and indirect immunofluorescence showed that rAdv-P12A3B3C-OZK93 expressed the FMDV-specific proteins P12A and VP1 in HEK-293 cells. In addition, the PK cell infection experiment confirmed that rAdv-P12A3B3C-OZK93 could infect porcine cells, which is essential for vaccination in pigs. Comparing with the inactivated vaccine group, the recombinant adenovirus could induce higher FMDV-specific IgG antibodies, γ-IFN and IL-10. This indicates that the recombinant adenovirus has good immunity for animal, which is very important for the subsequent development of foot-and-mouth disease vaccine.

Preparation and Quality Evaluation of Carbinoxamine Maleate Sustained-release Suspension / 中国药房

OBJECTIVE:To prepare Carbinoxamine maleate sustained-release suspension,and evaluate its quality. METH-ODS:Using carbinoxamine maleate as raw material,drug-loaded resin was prepared by cation exchange resin;surface coating method was used to finally prepare sustained-release suspension,using Eudragit RS100 as sustained-release coating material to pre-pare sustained-release microparticles. HPLC was conducted to determine the content of carbinoxamine maleate,release degree of original preparations and self-made suspensions was compared,drug-loading capacity was calculated. RESULTS:The drug amount in preparing drug-loaded resin was 2%,reaction temperature was 25 ℃,and reaction time was 4 h;the drug-loading capacity in surface coating was 35%,amount of coating material was 10%,and reaction temperature was 40 ℃. The drug-loading capacities of sustained particles before and after coating were 35.23%,32.72%,respectively;the yield was 96.82%. The carbinoxamine ma-leate in prepared sustained-release suspension accounted for 98.76% of the labeled amount;release degree in 10 h reached about 80%,f2 was 65.73. CONCLUSIONS:Carbinoxamine maleate sustained-release suspension is prepared successfully,and its release is similar to the original preparation.

Study of molecular characteristics of methicillin-resistant Staphylococcus aureus in Maanshan area / 中华流行病学杂志

Objective To identity the distribution of enterotoxin and hemolysin,as well as the clonal complexes and drug resistance of the strains of methicillin-resistant Staphylococcus aureus (MRSA) in Maanshan region.Methods Automatic enzyme-linked fluorescent assay system and PCR technology were used to identify the distribution of enterotoxin and hemolysin genes.Seven Staphylococcus aureus hourskeeping genes were choosed as the target genes for multilocus sequence typing (MLST) on 34 strains of MRSA and 3 strains of methicillin-sensitive Staphylococcus aureus (MSSA),comparing the data with the online database and obtaining the sequence typing (ST),conducting affinity analysis on its ST based on eBURST,testing in agar dilution method the drug resistance of MRSA against 12 antibiotics.Results 50.9％ of the 210 Staphylococcus aureus strains were enterotoxin positive,and 97.1％ of them carried hemolysin genes as all 51 strains of MRSA carried hemolsin genes.The 34 MRSA strains were divided into 10 STs,ranging in sequence ST239 (47.1％,16/34),ST5 (17.6％,6/34).Three MSSA strains belonged to ST188,ST1281 and ST7,respectively.Seventeen strains from the patients were divided into 6 STs,ranging in sequence ST239 (35.3％,6/17) and ST5 (29.4％,5/17).Twenty strains from food sources were divided into 9 STs,ranging in sequence ST239 (45.0％,9/20) and ST7 (15.0％,3/20).STs of ST585,ST630 and ST239 were close in affinity,while the rest were distant in affinity.Except for vancomycin,all the strains were found with drug resistance to varying extent to the 10 antibiotics tested.Conclusion Existence of Staphylococcus aureus hemotoxin was universal; ST239 was the main predominant MRSA in Maanshan region,with distant affinity among the STs.

Optimizing the preparation of doxycycline hydrochloride sustained release tablet using response surface methodology / 中国生化药物杂志

Objective In this article Response Surface Analysis(RSA)was applied to optimize the formulation of doxycycline hydrochloride sustained release tablet.Methods Single factor exploration was used to determine the three factors which have the greatest impact on the release rate.The three factors were the dosage of the HPMC in the total weight of the tablet,the concentration of PVP-K30,and the ratio of lactose to microcrystalline cellulose,respectively.The composite score of the release behaviour was taken as the response value.The dosage of the ingredients were determined by Box-Benhnke design principles and 3 factors and 3 levels.Results The optimized formulation and process are as follows:the dosage of the HPMC in the total weight of the tablet was 30%;the concentration of PVP-K30 was 10%,and the ratio of lactose to microcrystalline cellulose was 13.The release behavior in vitro is ideal.Conclusion The optimized preparation process of doxycycline hydrochloride sustained release tablet is stable,highly efficient and suitable for industrial production.

The In Vivo and In Vitro research status and future prospects of HPMC capsules / 中国生化药物杂志

After a literature review of the HPMC capsules, the research status of the HPMC capsules in vivo and in vitro are summarized, including the application status, the superiority over hard gelatin capsules and in particularly the disintegration release in vitro and bioavailability in vivo, as well as pharmacokinetics difference compared with conventional gelatine capsules, are explored in depth. Finally, the future applications of HPMC capsules are prospected.

Dissolution improvement of poorly water-soluble drug by cogrinding method using jar mill

As a representative proton pump inhibitor, Lansoprazole was poorly soluble in water which caused the low oral bioavailability. The present study was carried out to enhance the dissolution of lansoprazole by cogrinding with some commonly used hydrophilic polymers [beta-CD, PVP, HPMC, L-HPC, CS, PEG and PVPP] in the weight ratio of 1:1 for 2 h in the jar mill. Samples of coground mixture, micronised drug, and physical mixture were characterized by XRPD, and DSC, the results showed that the drug crystallinity reduced in the coground process. The amount of drug released from the coground mixtures in PBS [pH 6.8, 37[degree sign] C] in 30 min was 100% approximately [except the coground mixtures prepared with VPP or PEG] while released from the micronised drug was just about 20%. Increasing the hydrophilicity and diminishing the size of drug particles by cogrinding were the main causes for enhancing the dissolution of the drug. The results of the stability study of lansoprazole in coground mixture showed that there were no significant changes in the drug content and dissolution characteristics 6 months later. It is clear that the cogrinding method described in the article is very effective for enhancing the dissolution of the poorly soluble drugs, and it is easy for industrialization, showing a strong potential for future applications

Synthesis and biological evaluation of 3-aromatic Shiff base-5-fluoroindol-2-one derivatives / 药学学报

Based on the structure of 5-fluoroindol-2-one and fragments from thirteen multi-target tyrosine kinase inhibitors which have been marketed or in the phase of clinical research, eleven 3-aromatic Shiff base-5-fluoroindol-2-one derivatives were designed and synthesized. Their structures were identified by 1H NMR, MS and elemental analysis. In vitro antitumor bioactivities evaluation was done by MTT method. It was shown that most of synthesized compounds had antitumor activities and compounds 1b, 1g, 1i and 1h were better than or equal to the antitumor activity of positive control.

Preparation and identification of the polyclonal antibody against the 3D polymerase of foot and mouth disease virus / 中国人兽共患病学报

To prepare the polyclonal antibody against the 3D polymerase of foot and mouth disease virus (FMDV), the 3D polymerase gene of this virus was amplified by PCR and doubly digested with BamH I and Nde I. Then, it were cloned into expression vector pET-30a(+) to obtain the recombinant plasmid pET-3D and this plasmid was transformed to E.coli BL21(DE3) with induction by IPTG.The target protein was identified and purified with SDS+PAGE, and the inclusion bodies were extracted. The purified target protein was used as antigen to immunize New Zealand rabbits to prepare the polyclonal antibody against 3D polymerase of FMDV, which was then characterized by indirect ELISA and Western blotting. As demonstrated by SDS-PAGE, the target protein with a molecular weight at 46 ku was expressed. The polyclonal antibody showed high affinity and obvious specificity and its titer was above 1:8 000. This polyclonal antibody may lay a foundation for the further studies on the biological functions and epitopes of the 3D polymerase of FMDV.

Synthetic rehabilitation treatment of congenital muscular torticollis / 中国康复理论与实践

@#ObjectiveTo investigate the relationship between the prognosis and both of treatment way and curative age of congenital muscular torticollis.Methods138 children accepted conservative and/or operative treatment according to different ages.A prospective study had been made. ResultsConservative treatment was given to 82 children,71 of them were <1-year-old. 49 of them were follwed up and had satisfactory outcome.7 patients who was >1-year-old had operations later because of asymmetry of head.63 children received operations.56 of them were followed up over one year,47 cases had good-looking,the outcome of other 9 was not satisfied.Conclusions Operative intervention was necessary for >1-year-old patients. Manipulation was suitable way to cure <1-year-old patients.It was the best operating time at the age of 1-5 years old.