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Jiang K.,Orthopedics Oncology Institute of Chinese PLA
Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology | Year: 2011

To construct, express and purify a human fusion protein, which is composed of a single-chain antibody fragment scFv that recognizes HER2 protein and an oligo-9-arginine, and to analyze the binding activity of the expressed fusion protein. Pairs of oligonucleotide primers were designed and used to amplify the scFv-9R. The fusion protein gene scFv-9R was then cloned into expression vector pQE30 and expressed in E.coli M15.Expressed protein was detected by SDS-PAGE and Western blot and purified by Ni-NTA chelating agarose. Then, the purified protein was refolded by dialysis and concentrated by ultrafiltration. The antigen-binding activity of the scFv-9R fusion protein was confirmed by ELISA, and the siRNA binding ability was confirmed by electrophoretic mobility shift assay (EMSA). HER2 scFv-9R encoding sequence was correctly cloned into the expression vector. The recombinant protein was insolubly expressed in E.coli M15 induced by IPTG. ELISA confirmed that it had specific antigen binding activity; EMSA assured that it had siRNA binding activity. The scFv-9R fusion protein can specially bind with both HER2 antigen and siRNA. Source

Chen X.,Orthopedics Oncology Institute of Chinese PLA | Yang T.-T.,Orthopedics Oncology Institute of Chinese PLA | Qiu X.-C.,Orthopedics Oncology Institute of Chinese PLA | Ji Z.-G.,Orthopedics Oncology Institute of Chinese PLA | And 7 more authors.
Cancer Biology and Therapy | Year: 2011

Aim: to screen the pulmonary metastasis-associated molecules of osteosarcoma and evaluate their functions concerning prognosis prediction. Results: Analysis detected 484 differentially expressed genes between the high metastatic subline, F5M2, and the low metastatic subline, F4. There were 1,257 genes differentially expressed between newly established high-metastatic sublines named Saos-2M2 and its parental cell line Saos-2. Furthermore, 16 commonly upregulated genes and 5 commonly downregulated genes were identified by clustering analysis. EREG and CHST2, two genes not previously described in osteosarcoma, were finally seen to be differentially expressed in all examined osteosarcoma cell lines and in samples between the different prognosis sample groups. Survival analysis also confirmed these two molecules could be used to predict the outcome of OSA patients. Methods: cDNA microarrray analysis has been applied to 2 pairs of osteosarcoma cell sublines with differential metastatic potentials to the lung. Immunohistochemistry and survival analysis have been performed to clinical samples of osteosarcoma patients. Conclusion: This work represents a rationale approach to the evaluation of microarray data and will be useful to identify genes that may be causally associated with metastasis. EREG and CHST2 will be likely considered as clinical molecular markers to predict the outcome of OSA. © 2011 Landes Bioscience. Source

Yu Z.,Orthopedics Oncology Institute of Chinese PLA | Zheng L.,Orthopedics Oncology Institute of Chinese PLA | Wang Y.,Orthopedics Oncology Institute of Chinese PLA | Zhang Y.,Orthopedics Oncology Institute of Chinese PLA | And 2 more authors.
European Surgical Research | Year: 2010

Objective: The purpose of this study is to evaluate the functional and radiological outcomes of patients with unstable displaced proximal humeral fractures treated with closed reduction and percutaneous pinning (CRPP) fixation. Methods: We retrospectively reviewed 87 cases of displaced (2-, 3- or 4-part fractures according to Neer classification) proximal humeral fractures treated with CRPP fixation in our center from September 2003 to September 2008. Sixty-four patients were followed up for a period ranging from 12 to 48 months (averaging 16.2 months) and evaluated for the functional and radiological outcomes by a series of standard questionnaire and measurement. Results: The fractures in all 64 patients were healed with an average time of 15.4 weeks (ranging from 12 to 43 weeks), and the mean interval between the operation and full functional exercise was 17.3 weeks (ranging from 14 to 38 weeks). At the final follow-up visit, no patients showed shoulder instability; the mean range of abduction motion was 157.1° (ranging from 70 to 180°). For all patients, no statistically significant difference in the functional outcomes was observed between their 6-month and final follow-up visits, nor in the radiological findings between their immediately postoperative and fi- nal follow-up examinations. Conclusion: CRPP fixation is a feasible treatment option for unstable displaced proximal humeral fractures, especially for 2- and 3-part fractures in elderly patients. Although technically demanding, it offers reliable stability without extensive soft tissue dissection, allowing the early painless range of motion. This technique could also promote bone healing, prevent ischemic osteonecrosis of the head of humerus and lead to few complications. Copyright © 2010 S. Karger AG, Basel. Source

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