1.Preparation and identification of polyclonal antibody against gastric cancer-related protein GCRG224
Yinqiao WU ; Gangshi WANG ; Mengwei WANG
Medical Journal of Chinese People's Liberation Army 1983;0(02):-
Objective To prepare a polyclonal antibody against gastric cancer-related protein GCRG224.Methods The thioredoxin/GCRG224 fusion protein was expressed in E.coli.A polyclonal antibody against GCRG224 was obtained by immunizing a rabbit with the purified GCRG224 protein.The titer and specificity of the antibody were determined by ELISA and Western-blot,respectively.Results The thioredoxin/GCRG224 fusion protein with relative molecular mass of 16.8kD was over-expressed in E.coli.The purity of expressed products directly purified from a denaturing polyacrylamide gel was about 100%.The polyclonal antibody against GCRG224 was obtained.The ELISA titer of antiserum against GCRG224 was about 1∶256 000.Western blot analysis showed that the antiserum could bind to the expressed fusion protein specifically.Conclusion The polyclonal antibody against GCRG224 has been successfully prepared,which lays the foundation for further study on the biological function and the possible role of the GCRG224 in the development of gastric carcinoma.
2.Expression of gastric cancer related gene GCRG213 in E.coli and purification of recombinant protein
Yinqiao WU ; Gangshi WANG ; Mengwei WANG
Medical Journal of Chinese People's Liberation Army 1983;0(05):-
Objective To express gastric cancer related gene GCRG213 by using thioredoxin fusion expression system, and to prepare human GCRG213 fusion protein. Methods GCRG213 cDNA with complete open reading frame was amplified by PCR from plasmid pGEM-T, and then was cloned into thioredoxin fusion expression vector pET102/D-TOPO. The recombinant plasmid was further transformed into E.coli BL21 strain. After induction with IPTG, the thioredoxin/GCRG213 fusion protein was expressed in E.coli. The product was obtained by means of direct purification from a denaturing polyacrylamide gel. Results SDS-PAGE analysis showed the thioredoxin/GCRG213 fusion protein with relative molecule mass of 29.4kDa was highly expressed. The thin layer gel scanning analysis showed that the yield of GCRG213 fusion protein was 28.7% of the total bacterial protein. The product was obtained with a purity of about 100% by means of direct purification from a denaturing polyacrylamide gel. Conclusion The thioredoxin/GCRG213 fusion protein was successfully expressed in E.coli and the product with high purity was obtained, which laid the foundation for the function research and antibody preparation hereafter.
3. Effects of trabecular metal augments for the reconstruction of Paprosky type Ⅲ acetabulum bone defects
Yinqiao DU ; Yonggang ZHOU ; Shang PIAO ; Wenming WU ; Haiyang MA ; Zhisen GAO ; Jingyang SUN ; Chong ZHENG ; Sen WANG
Chinese Journal of Surgery 2017;55(6):410-415
Objective:
To investigate the methods and short-time clinical results of reconstruction of Paprosky type Ⅲ acetabulum bone defects by using tantalum augments.
Methods:
A total of 17 patients (17 hips) with Paprosky type Ⅲ acetabulum bone defects, treated with tantalum augments in revision of total hip arthroplasty at Department of Orthopedics Surgery in General Hospital of Chinese People′s Liberation Army were retrospectively analyzed from March 2014 to May 2016.There were 6 males and 11 females aged from 23 to 74 years with an average of (50.2±16.3) years. Tantalum augments or TM-Cup augment (the cup-on-cup technique) were used to reconstruct the defects.The TM-Cup augment was the tantalum revision cup which was removed titanium ring. The cup-on-cup technique combined TM-Cup augment and biological acetabulum cup. Augments were served as the nonresorptive structural allograft in revision of total hip arthroplasty. Harris hip score was used to evaluate clinical effects. The vertical position of the rotation center was measured and analyzed. Radiographic assessments of the acetabular components were performed by DeLee-Charnley and the Anderson criteria and recorded postoperative complications.
Results:
All the patients were followed up from 3 to 29 months with an average of (16.2±5.4) months, tantalum augments and biological acetabulum cup were used in 13 patients, the TM-Cup augment and biological acetabulum cup were used in 4 patients. At the time of the latest follow-up, the mean Harris hip score increased compared to preoperatively (86.8±8.3