Objective To construct the prokaryotic expression and purification protocols for Pseudomonas aeruginosa type E flagellin (FlgE) and to study its bioactivity.Methods With analysis of Pseudomonas aeruginosa flagellin FlgE sequences,the whole-length FlgE gene was amplified from Pseudomonas aeruginosa genomic DNA by using PCR and primers with proper restriction enzyme sites.The amplified FlgE fragment and prokaryotic expression plasmid pET24a were digested with Nde Ⅰ and Hind ⅢⅣ respectively.The target fragment and vector were recovered and ligated to obtain the recombinant plasmid pET24a-FlgE.DNA sequencing of positive clone confirmed that the target gene and the junctions with vectors were all correct.The plasmid pET24a-FlgE was transformed into BL21 bacteria.The culture conditions like temperature,rotation speed,inducer concentrations,time length were optimized to achieve maximal expression of the target recombinant FlgE with 6×His tag at C terminal.FlgE-His proteins were purified using His-Trap affinity chromatography columns and identified by SDS-PAGE.The purified proteins were further subjected to endotoxin elimination with proper kits.The purified recombinant FlgE was added to cultured corneal epithelial cells for 4 h and the expression of several inflammation-related molecules was examined by using real-time quantitative PCR.Results The recombinant plasmid pET24a-FlgE was successfully constructed and high level FlgE expression was achieved in BL21 with rotation at 16℃ and 1 mmol/L isopropyl-β-D-glucopyranosyl-galactosidase induction for 20 h.Purified recombinant FlgE-His was obtained and used for primary bioactivity assay.After treatment of corneal epithelial cells with 20 μg/ml FlgE for 4 h,the expression of inflammatory cytokines IL-6,IL-8 were significantly increased.Inactivation of the FlgE with ethanol abolished its stimulatory activity.Conclusion The prokaryotic expression and purification system for recombinant Pseudomonas aeruginosa flagellin FlgE was set up,and the recombinant FlgE stimulated the expression of inflammatory factors in corneal epithelial cells.

Defensin is a kind of antimicrobiol and cytotoxic peptides which have been found in a large range of living organisms. The mammal defensins have the most wide range antimicrobiol spectrum. The distribution, structure, gene expression regulation of mammal defensins in China and abroad were summarized. Its application on medicine and plant resistant gene engineering were prospected.

Staphylococcus aureus,an important foodborne pathogen,can contaminate foods through variety of ways and produce enterotoxin that may cause Staphylococcal food poisoning.In addition to food safety problems,Staphylococcus aureus could also cause clinical infection.The study of its pathogenicity is not only beneficial to prevent and control foodborne diseases,but also provide a new point for clinical treatment.This paper analyzes the types and characteristics of common strains of Staphylococcus aureus in food,and summaries the effect of food processing on pathogenicity and the methods for pathogenicity research in order to provide reference for the related study on pathogenicity of Staphylococcus aureus.

Objective: To study the effects of ginger oleoresins(GO) on blood lipids in hyperlipidemic mice and the platelet aggregation in rats.Methods: The rat platelet aggregation was induced by ADP and AA in vitro, and the inhibition by GO was tested by light transmission. The animal model of hyperlipidemia was establised by feeding Kunmimg mice with high lipid feed stuff. The effects of GO on serum lipid and hepatic MAD in hyperlipidemic mice was investigated. Results: GO could effectively inhibit rat platelet aggregation induced by ADP and AA in vitro, which were 58.9%,97.5% respectively, and in dose dependant manner. Besides, GO could also significantly decrease serum TC, LDL, AS index and hepatic MDA in hyperlipidemic mice, which were 16.45%, 60.45%,37.30% and 54.27% respectively, and showed no side-effect on liver. Conclusion: Long-term consumption of GO can effectively modulate blood lipids, prevent thrombosis and AS.

An adult patient with skeletal crossbite and mandibular deviation was treated by mandibular molar distalization using micro-im-plant and intermaxillary traction.After treatment,the facial contour of the patient was improved,the Class Ⅰ molar and cuspid relationship was achieved.

The anterior cruciate ligament (ACL), anterolateral complex (ALC) and lateral meniscus (LM) maintain the anterolateral rotatory stability of the knee and control the internal rotation of the tibia. Anterolateral rotatory instability (ALRI) of the knee is not uncommon in clinic, and its main injury mechanism is non-contact injury. A pivot shift test or a tibial internal rotation test can indicate ALRI while X-ray, CT, MRI and ultrasound can assist in its diagnosis and differential diagnosis. For acute ALRI, good technique of ACL reconstruction is the basis to avoid postoperative residual ALRI, and anterolateral ligament reconstruction and extra-articular tenodesis are optional as appropriate. For chronic cases, however, both anterolateral ligament reconstruction and extra-articular tenodesis are effective. This article reviews the progress in research on the diagnosis and treatment of ALRI of the knee, hoping to provide references for its clinical diagnosis and treatment.

To construct a three-dimensional finite element model comparing between one-step and two-step methods in torque control of anterior teeth during space closure.Methods Dicom image data including maxilla and upper teeth were obtained though cone-beam CT.A three-dimensional model was set up and the maxilla,upper teeth and periodontium were separated using Mimics software.The models were instantiated using Pro/Engineer software,and Abaqus finite element analysis software was used to simulate the sliding mechanics by loading 1.47 Nforce on traction hooks with different heights(2,4,6,8,10,12 and 14 mm,respectively) in order to compare the initial displacement between six maxillary anterior teeth (one-step method) and four maxillary anterior teeth(two-step method).Results When moving anterior teeth bodily,initial displacements of central incisors in two-step method and in one-step method were 29.26× 10-6 mm and 15.75× 10-6 mm,respectively.The initial displacements of lateral incisors in two-step method and in one-step method were 46.76× 10-6 mm and 23.18× 10-6 mm,respectively.Under the same amount of light force,the initial displacement of anterior teeth in two-step method was doubled compared with that in onestep method.The root and crown of the canine couldn't obtain the same amount of displacement in one-step method.Conclusions Two-step method could produce more initial displacement than one-step method.Therefore,two-step method was easier to achieve torque control of the anterior teeth during space closure.