Proteomic identification of necrozoospermia-related proteins / 中华男科学杂志

<p><b>OBJECTIVE</b>To compare the proteomic differences between normal and necrozoospermic human spermatozoa.</p><p><b>METHODS</b>We isolated proteins from 30 ejaculates of 10 healthy donors and 3 specimens of a necrozoospermia patient by two-dimensional gel electrophoresis (2-DE), analyzed their 2-DE maps and identified some differentially expressed proteins by mass spectrometry.</p><p><b>RESULTS</b>A total of (905 +/- 57) and (792 +/- 28) proteins were isolated from the spermatozoal maps of the normal and necrozoospermic men, respectively, and 178 found to be differentially expressed in that of the necrozoospermia patient. The 6 missing protein spots in the necrozoospermic map were identified as sperm protein (human, accession No. 060904), zinc finger protein 174 (AW-1), F-actin capping protein alpha-3 subunit, testis-specific inhibitor of apoptosis, death domain receptor 3 soluble form (fragment) and peptide similar to the activator of CREM in the testis.</p><p><b>CONCLUSION</b>Six missing protein spots were identified in the necrozoospermic spermatozoal map which may be associated with the development of necrozoospermia.</p>

Establishment of a high-resolution 2-D reference map of human spermatozoal proteins from 12 fertile sperm-bank donors / 亚洲男科学杂志(英文版)

<p><b>AIM</b>To extend the analysis of the proteome of human spermatozoa and establish a 2-D gel electrophoresis (2-DE) reference map of human spermatozoal proteins in a pH range of 3.5-9.0.</p><p><b>METHODS</b>In order to reveal more protein spots, immobilized pH gradient strips (24 cm) of broad range of pH 3-10 and the narrower range of pH 6-9, as well as different overlapping narrow range pH immobilized pH gradient (IPG) strips, including 3.5-4.5, 4.0-5.0, 4.5-5.5, 5.0-6.0 and 5.5-6.7, were used. After 2-DE, several visually identical spots between the different pH range 2-D gel pairs were cut from the gels and confirmed by mass spectrometry and used as landmarks for computer analysis.</p><p><b>RESULTS</b>The 2-D reference map with pH value from 3.5 to 9.0 was synthesized by using the ImageMaster analysis software. The overlapping spots were excluded, so that every spot was counted only once. A total of 3872 different protein spots were identified from the reference map, an approximately 3-fold increase compared to the broad range pH 3-10 IPG strip (1306 spots).</p><p><b>CONCLUSION</b>The present 2-D pattern is a high resolution 2-D reference map for human fertile spermatozoal protein spots. A comprehensive knowledge of the protein composition of human spermatozoa is very meaningful in studying dysregulation of male fertility.</p>

Diagnosing achondroplasia by single cell nested-PCR / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To research on the reliability of diagnosing achondroplasia (ACH) on single cell level and to provide a basis for preimplantation genetic diagnosis(PGD).</p><p><b>METHODS</b>The high-frequency mutation region G380R of fibroblast growth factor receptor 3(FGFR3) gene was amplified by nested-PCR with single lymphocyte and single blastomere. The products of PCR were digested by restriction enzyme Bfm I, then the digested products were detected by 10% polyacrylamida gel electrophoresis(PAGE).</p><p><b>RESULTS</b>The amplification success rate, allele dropout rate and correct diagnosis rate of single lymphocyte's PCR were 90.4%, 8.2% and 91.8%,respectively. The amplification success rate of single blastomere was 75.4%.</p><p><b>CONCLUSION</b>The diagnosis of ACH by single cell nested-PCR is comparatively stable and reliable.</p>