Nuclear matrix proteins differentially expressed in human prostate cancer cell lines and benign prostatic hyperplasia epithelial cell line / 中华男科学杂志

<p><b>OBJECTIVE</b>To compare the expression of nuclear matrix proteins (NMPs) in benign prostatic hyperplasia (BPH) epithelial cell line BPH-1 versus those in androgen-dependent human prostate cancer cell line LNCap and androgen-independent prostate cancer cell line PC-3.</p><p><b>METHODS</b>We isolated NMPs from the BPH-1, LNCap and PC-3 cell lines by 2-dimensional electrophoresis (2-DE), analyzed the differentially expressed proteins by matrix-assisted laser desorption / ionization time of flight mass spectrometry (MALDI-TOF-MS), and identified them by peptide mass fingerprint and database searching.</p><p><b>RESULTS</b>We successfully obtained well-resolved reproducible 2-DE patterns of NMPs in human prostate cancer cell lines, identified 12 differentially expressed NMPs including enzymes, regulatory proteins, RNA-binding protein and various other factors, 3 up-regulated and 9 down-regulated in prostate cancer cell lines.</p><p><b>CONCLUSION</b>There are obvious differences in the expressions of NMPs between human prostate cancer cell lines and benign prostatic hyperplasia epithelial cell line.</p>

Identification of M(r)22 000 protein in rat epididymal luminal fluid / 中华男科学杂志

<p><b>OBJECTIVE</b>To identify the specific protein in the epididymal luminal fluid that may play a role in sperm epididymal maturation or modification on the surface of spermatozoa.</p><p><b>METHODS</b>We compared the differential protein components in the lumen fluids from the caput and cauda segments of the epididymis of normal rats as well as from the cauda segment of experimental left varicocele (ELV) rats by SDS-PAGE or 2D-electrophoresis. The protein spots of interest were selected for MS identification, and the target proteins further characterized by immuno-blot assay.</p><p><b>RESULTS</b>MS analysis showed that one of the most prominent proteins, M(r) 22 000, was identical to the phosphatidylethanolamine-binding protein (PBP), and it was further identified as PBP by immuno-blot assay.</p><p><b>CONCLUSION</b>PBPs were present in a variety of molecular forms in the epididymal luminal fluid, including the glycosylated form, and ELV markedly elevated the PBP level in the cauda luminal fluid of the rats. Thus, the association of this molecule with sperm surface modification remains an interest for future investigation.</p>

Specific expression of beta-actin during spermatogenesis in rats / 中华男科学杂志

<p><b>OBJECTIVE</b>To screen the stage-specific expression proteins during rats spermatogenesis, and to investigate the beta-actin expression and localization in the tissues of rat testicular.</p><p><b>METHODS</b>Highly enriched type A spermatogonia, pachytene spermatocytes and round spermatids were isolated by STAPUT method (sedimentation velocity at unit gravity, with 2% - 4% BSA gradient in DMEM/F12 medium) respectively to get the total proteins. The difference of protein expression between the three kinds of cells was analyzed by two-dimensional electrophoresis. Then the distribution of beta-actin in rat testicular tissues was investigated using specific anti-beta-actin antibodies by immunohistochemical method.</p><p><b>RESULTS</b>beta-actin was identified as a stage-specific expression protein by two-dimensional electrophoresis. beta-actin protein was more strongly expressed in type A spermatogonia and pachytene spermatocytes, but not in round spermatids. The immunohistochemical results showed that beta-actin was mainly located in the cytoplasm of type A spermatogonia and pachytene spermatocytes and in the nuclei of nearly mature spermatids.</p><p><b>CONCLUSION</b>beta-actin protein is a stage-specific expressed protein and may play an important role in spermatogenesis.</p>

Effect of experimental varicocele on structure and function of epididymis in adolescent rats / 亚洲男科学杂志(英文版)

<p><b>AIM</b>To study the effect of experimental left varicocele (ELV) on epididymal structure and function in adolescent Sprague-Dawley rats.</p><p><b>METHODS</b>ELV was induced by partial ligation of the left renal vein. Sham-operated animals served as the controls. Four and 8 weeks after the operation, the histological, ultrastructural and biochemical (alpha-glucosidase activity and carnitine content) changes in different segments of the epididymis were observed.</p><p><b>RESULTS</b>In the treated animals, there were degeneration of the epididymal epithelium and edema of the interstitial tissue; numerous shedding cells, residual bodies, deformed sperm and macrophages appeared in the epididymal lumen. Morphometric measurement indicated a significant reduction in the epididymal tubular diameter (P<0.05) and a significant increase in the epididymal interstitial area (P<0.05) compared with the controls. Ultrastructural study showed sparse microvilli of the columnar epithelium, increased and enlarged lysosomes in the principal cells with defected organelles and the presence of large cytoplasmic vacuoles. The protein and carnitine contents and the alpha-glucosidase activity in the caput, corpus and cauda epididymis of the ELV rats were lower than those of the controls (P<0.05).</p><p><b>CONCLUSION</b>There were structural and functional changes in the epididymis of adolescent ELV rats, which may contribute to the infertility caused by varicocele.</p>

Differential protein analysis in rat renal proximal tubule epithelial cells in response to acetazolamide and its relation with the inhibition of AQP1 / 药学学报

<p><b>AIM</b>To study the endogenous mechanism for the inhibition of aquaporin-1 expression in rat renal proximal tubule epithelial cells in response to acetazolamide.</p><p><b>METHODS</b>Primary cultured rat renal proximal tubule epithelia cells were divided into two groups: one was subjected to 1 x 10(-5) mol.L-1 acetazolamide, the other served as normal control. When grown to sub-confluency, the cells were disintegrated to perform isoelectrofocusing electrophoresis in order to find the differential proteins induced by the acetazolamide treatment. The differential proteins were defined by peptide mass fingerprinting technology.</p><p><b>RESULTS</b>Two differential proteins were found in the cell disintegrant. The pI 3.8 protein was reduced after treatment, which showed 21.4% similarity with the brush border membrane myosin from rat brain and testis, and 27% with glycogen phosphorylase; The pI 5.5 protein was increased on the contrary, with 20% similarity to phosphatidylinositol transfer protein alpha isoform.</p><p><b>CONCLUSION</b>Acetazolamide inhibited AQP1 expression probably by affecting the expression of pI 3.8 and pI 5.5 proteins.</p>