ABSTRACT
Objective:To prepare monoclonal antibody ( mAb ) against human testis-specific conserved gene ( hTSC29 ) peptides and characterize its immunological and biological features.Methods:According to bioinformatics analysis and prediction of the antigenicity, surface property, hydrophilicity and flexibility of hTSC29, a 18-amino acid residue partial peptide of hTSC29 was synthesized,then immunized the BALB/c mice for preparing antiserum.The mAb against hTSC29 was produced using the routine hybridoma technique.The properties of the mAb against hTSC29 were identified by ELISA, Western blot and immunohistochemistry staining.Results:After cell fusion and subcloning, one hybridoma cell lines secreting specific mAb against hTSC29 protein were obtaind.The Ig subclass of the mAb was IgG2b(κ).ELISA detection showed that the titer of mAbs in cultured was 1∶104.Western blot analysis proved that the mAb could specifically recognize Mr 60 000 protein in human testis total protein.The hTSC29 protein main located at circumference of spermatocyte and spermatid in human testis tissue by immunohistochemistry staining and immunofluorescence assay.Conclusion:One hybridoma cell lines which can secrete specific mAb against hTSC29 protein with high titers and specificity have been established successfully.The mAb will provide efficient tools for functional studies of hTSC29 expressed in spermatogenesis.
ABSTRACT
Objective To study the relationship between the semen quality and age in infertile men in Qingyuan.Methods 727 cases of infertile men from Reproductive Medicine Center outpatient were enrolled and divided into 5 groups:≤ 25,> 25 - 30,>30-35,>35-40,>40 age groups.The semen characteristics were carried out by calculator assistance sperm analysis(CASA) technique and Diff-Quick staining method.Results The sperm density of >30 -35 age group was the highest,but there was no significantly difference from other groups(P >0.05).Semen analysis showed that the rates of PR,PR+NP and survival sperm were on downward trends with the increase of age,but not statistically significant(P >0.05).38.19% of >25 -30 age group reached WHO′s semen parameters standard,which was the highest proportion among 5 groups.And the proportion in >40 age group was the lowest,which counted for 27.42%.Conclusion The semen quality shows a down trend with the increase of age,however,the infertility could occur in the reproductive men at any stage.