Objective: To fuse Mycobacterium tuberculosis protective antigen gene with mice ubiquitin gene, constructing antigen ubiquitin system. Methods: Mice ubiquitin cDNA was amplified by RT PCR from mice testicle,and 4 antigen genes were obtained by PCR from cultured Mycobacterium tuberculosis . Ubiquitin and 4 antigen genes were linked by flexible adaptor respectively and the fusing genes were cloned into pVAX vector.The recombinant plasmids were digested with endonuclease and sequenced.Then the recombinant plasmids were transfected into COS7 cells and the expression was assayed by ELISA. Results:Ubiquitin and 4 antigen genes were 0.2,0.3,0.7,1.0,1.65 kb in length by agarose electrophoresis. Endonuclease digestion of the recombinant plasmids indicated that the fusion genes were correctly inserted into pVAX vector. Sequencing results of fusion nucleic acid vaccines were identical to those in GenBank.The recombinant plasmids expressed in COS7 cells. Conclusion: Four Chinese Mycobacterium tuberculosis protective antigens ubiquitin systems are successfully constructed and can be expressed in eukaryotic cells. This may provide a basis for diagnosis and therapy of tuberculosis.

Objective To investigate the morphological and immunophenotypic characteristics of phyllodes tumors (PT) and explore their significance in differential diagnosis and evaluation of prognosis.Methods 48 specimens with PT were observed under light microscope with HE and immunohistochemical staining (EnVision method).The antibodies including CD34,CD10,CD117,p53,and Ki-67 were used.10 cases of fibroadenoma were compared and the clinical data and follow-up results were analyzed retrospectively.Results All of 48 cases with PT presented as well-circumscribed masses with typically leaflike structures composed of double-layered epithelial component arranged in clefts and overgrowing hypercellular mesenchymal component.31 benign,12 borderline,and 5 malignant PT were diagnosed based on stromal overgrowth,cellular pleomorphism,mitosis,margins,and others.Focal necrosis was detected in 3 malignant cases and liposarcoma component existed in 1 case.Immunohistochemical staining showed that the positive rates of CD34 were 93.5 ％,58.3 ％,0 in benign,bordline,and malignant PT respectively,there were significant differences between each groups (P ＜ 0.05).The expression of CD10 were 25.8 ％,83.3 ％,80.0 ％,its expression in benign group was significantly different compared with bordline and malignant PT (P ＜ 0.05).p53 was expressed in three groups with no significant difference (P ＞ 0.05).CD117 and Ki-67 had high expression comparatively in malignant PT.All cases were treated surgically with a local recurrence rate of 22.9 ％.Conclusions Reasonable surgery pattern based on accurate histopathological diagnosis is crucial to reduce the local recurrence rate of PT.Immunohistochemistry examination in PT is helpful to the differential diagnosis and evaluation of prognosis.

BACKGROUND:Generaly, the stent surface modification, especialy seeding cels, may accelerate or cause stent endothelium, and cause restenosis for prevention of in-stent thrombosis.
OBJECTIVE: To develop the optimal conditions for vascular stents coated with bone marrow mesenchymal stem cels.
METHODS: Cerebrovascular stent was co-cultured with passage 3 bone marrow mesenchymal stem cels from rats at 1×106, 1×107, 1×108, and 1×109/L. Cels on the stents were examined with transmission electron
microscopy after 48 hours. A total of 160 male Sprague-Dawley rats were enroled, among which, 20 rats were as normal control group, and the remaining 140 were used for producing models of ischemic stroke that were
randomly sub-divided into seven groups at 8 weeks after modeling: stainless steel stent implanted group, polymer stent group, and different concentrations of cellstent composite groups. After 8 weeks of implantation, the
expression of vascular endothelial growth factor in these cels was examined by western blot assay. Rat platelet activation in different groups was determined by flow cytometry.
RESULTS AND CONCLUSION:Implanted stem cels were able to grow adherently on the stainless steel stent wal. When the planting cellconcentration was 1×107 cels/L, the cels and organeles were morphologicaly
normal and covered the stent surface wel. These coated cels also expressed vascular endothelial growth factor, suggesting that they functioned as endothelial cels, and they also significantly lowered platelet activation. When
co-cultured with 1×107/L bone marrow mesenchymal stem cels, the stent was covered wel with endothelial-like cels and had significant lower platelet activationin vivo.

Objective To analyse the immunostimulatory activity of CpG sequences in cysticercus cellulosae paramyosin (also named Antigen B,AgB)cDNA. Methods C57BL/6 mice were immunized with pcDNA3 AgB plasmid,pcDNA3 AgB′(CpG sequences were mutated),pcDNA3 or AgB protein and two weeks later,immune response was assayed by ELISA. Results IgG and IgG 2a were detectable at week 2 after immunization and continually increased until week 4.The antibody levels elicited by pcDNA3 AgB were significantly higher( P

Objective To clone and express the gene encoding ESAT antigen of Mycobacterium tuberculosis (chinese stain). Methods Genome of mycobacterium tuberculosis was extracted. Then, the full length cDNA encoding ESAT protein was amplified by PCR and cloned into prokaryotic expressing vector pGEX5T and sequenced. pGEX5T ESAT was expressed in k802 Ecoli, and the expressed protein was determined by western blot using the sera from ten patients with tuberculosis. Results One specific band of 0.3kb or so was obtained and sequencing result was identical to that reported from Genbank. The expressed protein could be specifically recognized by the sera from tuberculosis patients. Conclusions The full length cDNA of Mycobacterium tuberculosis (Chinese strain) ESAT protein was cloned and expressed successfully, which will be helpful in further studies on diagnosis and treatment of tuberculosis.

Objective To test the biomechanical properties and evaluate the feasibility of the new developed external fixator system,so as to provide reliable theoretical basis for further clinical research.Methods The tibia of 10 fresh male cadaver were made into fracture model.In order to fix the fracture of tibia,experimental group were treated with the new developed external fixator system and the control group were treated with the Hoffman 2 external fixator which was used in clinic commonly.Then axial compression and horizontal torsion test were made on tibia fracture model.The test results were recorded and statistically analyzed.Results There was no statistical difference on the aspect of the ability to resist axial compression and horizontal torsion between the experimental group and the control group(P >0.05). The new developed external fixator system could meet the criterion in clinical application.Conclusion The new developed external fixation system can meet the requirement of biomechanics on fixing the fracture of bone,and it can greatly simplify and diversify the clinical applica-tion of external fixation device.

Objective To develop a newly designed guide and evaluate its clinic application prospect in treatment of patella fracture. Methods In our mechanical experiment, we carried out the tensile test of the two generation drilling guides by biomechanics machine and then analyzed the difference in their deformation. In the corpse experiment , we transversed fractures at the equator of the patella obtained from 10 fresh cadaver speci-mens. They were divided into 2 groups randomly: a test group and a control group. The second-generation of the aiming guide was used for the test group but not for the control group during the experiment. Then we evaluated its clinic application in the treatment of patella fracture. Results In the mechanical experiment, the displace-ment of the first generation guide was significantly larger than that of the newly one. The accuracy in the place-ment of k-wires by the second generation guide was significantly higher than by the first generation guide (P <0.05). Conclusions The drilling guide is reasonable in design and convenient in use. Moreover, it can improve the accuracy and quality of insertion of tension-band wiring of patellar fractures. The second-generation one is better and thus it is worth clinically spreading.

AIM: To quantify magnesium isoglycyrrhizinate(MGL) and glycyrrhetic acid in the plasma of dog by develop a simple, rapid, sensitive high-performance liquid chromatography (HPLC-UV) method. METHODS: HPLC-UV methods with wavelength 252 nm were used for the quantitation of MGL and GA in plasma. The concentration of MGL and GA were assayed on a Kromasil ODS-1 C18 column with the column temperature 25 ℃. The mobile phase was a gradient system with 0.1 % diethylamine in water (pH 4.60 ) and acetonitrile at a flow rate of 1.0 ml?min~ -1 . RESULTS: There was a good linear response range of 0.2 -2.5 mg?L~ -1 and 2.5 -100 mg?L~ -1 , while the limit of quantification was 0.2 mg?ml~ -1 . The relative recovery rate of MGL was 94.3 %-101.9 %,GA 96.4 %-101.9 % (n=5);the absolute recovery rate was MGL 78.7 %-87.0 %, GA 77.5 %-87.7 % (n=5). The intra-day and inter-day variations were all less than 15% (n=5). The plasma samples preserved in refrigerator is stable at -20 ℃ for 14 days, freeze thawing 3 times and at room temperature for 10 h without degradation. CONCLUSION: This method is shown to be specific, sensitive, reliable and suitable for the quantitative determination of magnesium isoglycyrrhizinate following oral administration in biological sample.

Objective To develop a new designed Kirschner wires guider for patellar fracture,and to evaluate its clinic outcomes.Methods From October 2010 to November 2015,totally 72 patients with patellar fracture were detected for this study.Divided them into 3 groups, named as the second-generation guider group,the first-generation guider group and the control group.And then evaluated the clinic outcomes in treatment of patellar fracture of the 3 groups.Results The surgery time,accuracy of placement needle position of the second-generation guider group were better than the other two groups with significant statistical differences (P <0.01).And the clinic outcomes of the first-generation guider group were better than the control group,and the differences were statistically significant (P <0.05).Conclusion The second-generation Kirschner wires guider is reasonable in design and convenient for use.Moreover,it can improve the accuracy and quality of surgery,reduce the operation time,and enhance the clinical effects.

Objective To explore an effective way for medical students and young surgeons learning operative approach .Methods A total of 50 medical students and young surgeons in our hospital were detected for this study .They were randomly divided into the BOOK class (group A) and the 3D computer class (group B) to learn the course of anthropotomy.After learning each section, they were examined to-gether on fresh corpse , and the examined results of the two groups were compared and assessed .Results In the general surgery examina-tions, there was no difference in the examined results with no statistically significant difference (P>0.05).But in the orthopedics examina-tions, the difference of intra-group comparison both in group A and group B were of statistical significance (P<0.05), and the results of group B were better than group A with statistically significant difference (P<0.05).Conclusion For the general surgical approach learn-ing, both of the two methods could achieve good effect .For the orthopedics surgical approach learning ,it can receive better results through computer software ,but it still cannot achieve the ideal effect ,which indicates that the teaching through fresh corpse could not be replaced yet .