Prokaryotic expression of a recombinant protein of adeno-associated virus capsid conserved regions and preparation of its polyclonal antibody / 南方医科大学学报

OBJECTIVE@#To express and purify the antigenic peptide of adeno-associated virus (AAV) capsid conserved regions in prokaryotic cells and prepare its rabbit polyclonal antibody.@*METHODS@#The DNA sequence encoding the conserved regions of AAV capsid protein was synthesized and cloned into the vector pET30a to obtain the plasmid pET30a-AAV-CR for prokaryotic expression and purification of the conserved peptides. Coomassie blue staining and Western blotting were used to identify the AAV conserved peptides. Japanese big ear white rabbits were immunized with AAV conserved region protein to prepare polyclonal antibody, with the rabbits injected with PBS as the control group. The antibody titer was determined with ELISA, and the performance of the antibody for recognizing capsid protein sequences of AAV1-AAV10 was assessed with Western blotting and immunofluorescence assay.@*RESULTS@#The plasmid pET30a-AAV-CR was successfully constructed, and a recombinant protein with a relative molecular mass of 17000 was obtained. The purified protein induced the production of antibodies against the conserved regions of AAV capsid in rabbits, and the titer of the purified antibodies reached 1:320 000. The antibodies were capable of recognizing a wide range of capsid protein sequences of AAV1-AAV10.@*CONCLUSION@#We successfully obtained the polyclonal antibodies against AAV capsid conserved region protein from rabbits, which facilitate future studies of AAV vector development and the biological functions of AAV.

Modified Bianchi orchiopexy for median or low cryptorchidism / 中华男科学杂志

Objective@#To investigate the effect of modified Bianchi (single incision in the midline of the scrotum) orchiopexy (MBO) versus that of traditional surgery in the treatment of median or low cryptorchidism.@*METHODS@#Eighty-two children with median or low cryptorchidism were treated from February 2013 to February 2014, 46 (53 testes) by MBO and the other 36 by the traditional method of inguinal incision (control, 40 testes). Comparisons were made in the operation time and postoperative complications between the two surgical strategies.@*RESULTS@#The mean operation time was significantly shorter in the MBO group than in the control (［25±6］ vs ［35±4］ min, P0.05). The incision scar was obvious in all the controls, with 1 case of postoperative inguinal hematoma, but almost invisible in all the MBO cases.@*CONCLUSIONS@#Modified Bianchi orchiopexy is superior to traditional surgery in the treatment of median or low cryptorchidism for its advantages of short operation time, few complications, and satisfactory appearance of the healed incision.