ABSTRACT
To prepare polyclonal antibodies (PcAb) against UspA1 of Moraxella catarrhalis (Mc), we used bioinformatic analysis to determine the surface exposed region in this protein that holds the antigen epitopes. Then the corresponding coding sequences for this fragment was artificially synthesized according to the codon usage of Escherichia coli. The gene fragment was then subcloned into the prokaryotic expression vector pET-28a(+) and expressed in E. coli rosseta (DE3), and then the recombinant UspA1-His proteins were purified. Two New Zealand white rabbits were immunized with this protein to prepare antiserum. The resulting PcAb was then purified from the antiserum with Protein A affinity column. The results of fluorescence antibody assay, enzyme linked immunosorbent assay and Western blotting analysis showed that the PcAb could specifically recognize the surface exposed region of UspA1 on Mc. The preparation of the PcAb laid a foundation of further development of rapid detection technique for M. catarrhalis.
ABSTRACT
@#Objective To investigate the expression of mutant BRCA2 gene in human breast cancer tissue and its clinic-healing significance.Methods The expression of mutant BRCA2 gene in breast cancer tissues and adjacent non-cancerous tissues of 66 patients' samples was detected using immunofluorostainning(IFS)and the correlation between the expression of mutant BRCA2 and patient's clinic-pathological characteristic was analyzed.Results Among 66 patients' samples,87.8%(58/66)were up-regulated expression of mutant BRCA2 protein in breast cancer tissues and 12.2%(8/66)was no significant difference of the mutant BRCA2 expression between the cancer tissue and its corresponding adjacent noncancerous tissues.The label index(LI)of mutant BRCA2 gene was higher in the cases of breast carcinoma's later clinical stage,poor tumor differentiation grade,breast scirrhus carcinomas,infiltrative breast carcinomas and metastasis.Conclusion The overexpression of mutant BRCA2 gene in human breast cancer tissue is positively related to the breast carcinoma's clinical stage,tumor differentiation grade,the tumor pathological type and metastasis,and it may influence the healing therapy of the breast cancer patients.
ABSTRACT
@#Objective To investigate the expression of poly ADP-ribose polymerase-1(PARP-1) in human breast cancer tissue and its relationship with patient's prognosis related factors.Methods The expression of PARP-1 in breast cancer tissues and their adjacent non-cancerous tissues from resection in 66 operated cases were detected using RT-PCR and immunofluorecence staining(IFS).The relationship between the expression of PARP-1 and the prognosis related factors was analyzed.Results Compared with that of the adjacent non-cancerous tissues,the expression of PARP-1 mRNA increased in 94.9%(62/66) patients,while the expression of PARP-1 protein increased in 90.9%(60/66) ones.The label index(LI) and the expression index(EI) of PARP-1 gene increased especially in the cases in later clinical stage,with poor tumor differentiation grade,of pure breast carcinomas or infiltrative breast carcinomas and of metastasis.Conclusion The expression of PARP-1 in human breast cancer tissue on the level of transcription and translation increases,which may be positively related with the patient's prognosis.