Clinical research of SRT and 3D-CRT on treatment of brain metastasis form lung cancer / 重庆医学

Objective To assess the evaluation of SRT (Stereotactic radiotherapy ) and 3D-CRT (Three-dimensional conformal ra-diotherapy)on treatment of brain metastasis from lung cancer .Methods From June 2009 to June 2011 ,74 patients with multiple brain metastasis(brain metastasis ≤3 ,tumor mass ≤3 cm) from lung cancer were analyzed retrospectively ,37 patients received 3D-CRT alone were retrospectively compared with 37 patients who received SRT alone .the results was evaluated by median survival time(MS) ,local control(LC) and toxicity effect .Results The median survival time(MS) was 9 .3 ,which 8 .6 months after 3D-CRT ,and 10 .6 months after SRT .the local control rate was 73 .5% for 3D-CRT ,and 79 .6% for SRT after 3 months .there was no difference between two groups on toxicity effect from stastic .Conclusion The SRT was a efficacious methods for the treatment of brain metastasis from lung cancer ,which could improve the local control rate ,but there was no more toxiticy .

Screening and primary identification of anti-CCR7 scFv from large phage library / 中国药理学通报

Aim To screen and identify anti-CCR7 single chain fragments variable(scFv)from lager phage library and to detect the scFv efficiency.Methods The insert ratio of ScFv antibodies library was identified by PCR.The products digested by Sfi I/Not I double enzyme.Panning against breast cancer cell and CCR7 were performed four and three rounds respectively.Positive clones were transformed to E.coli HB2151, and their dissolvability was assayed.The soluble scFv was purified by affinity chromatography, and its relative molecular mass was determined by Western blot.The ELISA assay was used to identify the immunocompetence of the antibody.Immunocytochemical staining and radioimmunoimaging were employed to determine the affinities of scFv binding with CCR7 in cell line and in nude mice.Results The insert ratio of ScFv gene was 90%(18/20), enzyme digest reaction showed the aim products on 1% agarose gel.ScFvs were obviously enriched after 7 round panning.Western blot result showed soluble scFv's molecular mass was about 34 KD.ELISA analysis showed dissolved antibody had high immunocompetence to MDA-MB-435 s cells.Immunocytochemical staining and radioimmunoimaging indicated that ScFvs were bound efficiently to MDA-MB-435 s cells which expressed CCR7.Conclusions ScFvs against CCR7 are successfully acquired by screening the phage antibody library.The soluble ScFvs have high affinity and specifical binding to human breast cancer cells.

Construction of human phage display antibody ScFv library and identification of antibody ScFv against lung adenocarcinoma / 基础医学与临床

Objective To construct a human phage single chain-antibody library, and to sieve out the antibody ScFv against lung cancer from the library. Methods Total RNA was abstracted from lymph node tissue of the lung cancer, and was used to amplify V_H and V_L gene by RT-PCR. V_H and V_L were joined by a DNA linker by SOE-PCR to form the single chain variable fragment ( ScFv) gene. ScFv gene was coloned into the phage vector pCANT-AB5E. Panning against lung cancer cell line A549 was performed and positive clones were chosen for soluble expression. Results A recombination phage single chain-antibody library was constructed. After 4 rounds panning, the number of eluted phages increased by 115 times. Positive reactions to A549 were detected in 7 of 10 random clones. The human ScFvs against lung cancer were produced and confirmed by SDS-PAGE and ELISA analysis. Conclusion ScFvs against lung cancer were acquired by the construction of phage single chain-antibody library. The soluble ScFvs has specificall avidity to human lung cancer cells.

Construction of human phage display antibody ScFv library and identification of antibody ScFv against lung adenocarcinoma / 基础医学与临床

Objective To construct a human phage single chain-antibody library,and to sieve out the antibody ScFv against lung cancer from the library.Methods Total RNA was abstracted from lymph node tissue of the lung cancer,and was used to amplify VH and VL gene by RT-PCR. VH and VL were joined by a DNA linker by SOE-PCR to form the single chain variable fragment (ScFv) gene. ScFv gene was coloned into the phage vector pCANTAB5E. Panning against lung cancer cell line A549 was performed and positive clones were chosen for soluble expression.Results A recombination phage single chain-antibody library was constructed. After 4 rounds panning,the number of eluted phages increased by 115 times. Positive reactions to A549 were detected in 7 of 10 random clones. The human ScFvs against lung cancer were produced and confirmed by SDS-PAGE and ELISA analysis.Conclusion ScFvs against lung cancer were acquired by the construction of phage single chain-antibody library. The soluble ScFvs has specificall avidity to human lung cancer cells.