Inhibition of bladder cancer cell growth and angiogenesis by co-blockage of vascular endothelial growth factor and its receptor KDR / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To study the effect of co-blockage of vascular endothelial growth factor (VEGF) and its receptor (KDR) on growth of bladder carcinoma T24 cells and nude mice xenograft.</p><p><b>METHODS</b>T24 cell line co-transfected with VEGF siRNA and sKDR expression plasmids was developed and its proliferation was assayed by MTT and apoptosis by FCM. The nude mice model bearing bladder carcinoma xenograft was established. The tumor cell VEGF expression, stroma microvessel density (MVD) and tumor cell topoisomerase II alpha (Topo II alpha) expression were detected by immunohistochemistry. Cell apoptosis was estimated by TUNEL assay.</p><p><b>RESULTS</b>MTT assay showed that cell proliferation in VEGF siRNA, sKDR and combination groups was 56.3% +/- 8.3%, 42.6% +/- 13.8% and 32.5% +/- 4.3%, respectively, significantly lower than that in the scramble control (97.3% +/- 11.6%, P < 0.0001). FCM showed there were sub-diploid apoptotic peaks before G1 phase in VEGF siRNA, sKDR and combination groups, and apoptosis ratio was 5.1% +/- 0.9%, 4.2% +/- 0.5% and 8.8% +/- 0.7%, respectively, all of which were higher than that in the scramble control (0.9% +/- 0.4%, P < 0.05), and the combination group had even more higher apoptosis than the two singlely treated groups (P < 0.01). In vivo test showed that tumor growth was inhibited in VEGF siRNA, sKDR and combination groups, and from day 16 the tumor volume in combination group was significantly smaller than that in scramble control (P < 0.05), and from day 28 the tumor almost lost the ability to further growth. Immunohistochemistry revealed VEGF expression in combination group was 54.37 +/- 5.28, significantly lower than that in the scramble control (141.66 +/- 8.59, P < 0.0001). MVD number was only 8.22 +/- 3.79, much less compared with that in the scramble control (61.76 +/- 5.28, P < 0.0001) or sKDR group (19.46 +/- 4.16, P = 0.0089). Tumor cell proliferation index in the combination group (1.5% +/- 0.7%) was significantly decreased compared with that in the scramble control (11.8% +/- 5.2%, P < 0.0001), and apoptosis index (67.2% +/- 8.5%) was much higher than that in the scramble control (8.7% +/- 2.7%, P < 0.0001), VEGF siRNA group (54.3% +/- 4.8%, P = 0.0492) or sKDR group (52.3% +/- 6.4%, P = 0.0293).</p><p><b>CONCLUSION</b>VEGF siRNA or sKDR alone can inhibit tumor cell proliferation and induce cell apoptosis, but co-blockage of VEGF and KDR by their combination shows more significant therapeutic efficacy.</p>

Assessment of released acrosin activity as a measurement of the sperm acrosome reaction / 亚洲男科学杂志(英文版)

<p><b>AIM</b>To develop a method for assessing sperm function by measuring released acrosin activity during the acrosome reaction (AR).</p><p><b>METHODS</b>Human semen samples were obtained from 24 healthy donors with proven fertility after 3-7 days of sexual abstinence. After collection, samples were liquefied for 30 min at room temperature. Standard semen parameters were evaluated according to World Health Organization (WHO) criteria. Calcium ionophore A23187 and progesterone (P4) were used to stimulate the sperm to undergo AR. After treatment, sperm were incubated with the supravital dye Hoechst33258, fixed in a glutaraldehyde-phosphate-buffered saline solution, and the acrosomal status was determined by fluorescence microscopy with fluorescein isothiocyanate-labeled Pisum sativum agglutinin (FITC-PSA). The percentage of sperm undergoing AR (AR%) was compared to sperm acrosin activities as assessed by spectrocolorimetry. The correlation between AR% and acrosin activity was determined by statistical analysis.</p><p><b>RESULTS</b>The AR% and released acrosin activity were both markedly increased with A23187 and P4 stimulation. Sperm motility and viability were significantly higher after stimulation with P4 versus stimulation with A23187 (P < 0.001). There was a significant positive correlation between released acrosin activity and AR% determined by FITC-PSA staining (r=0.916, P < 0.001).</p><p><b>CONCLUSION</b>Spectrocolorimetric measurement of released acrosin activity might serve as a reasonable alternative method to evaluate AR.</p>

Relationship between sperm motility parameters and sperm morphology / 中华男科学杂志

<p><b>OBJECTIVE</b>To evaluate the relationship between sperm motility parameters and sperm morphology.</p><p><b>METHODS</b>Seven hundred and eighty-three semen samples were tested. Sperm motility parameters were analyzed by computer-aided sperm analysis (CASA) , and sperm morphology assessed by automated sperm morphology analyzer (ASMA). The cases were classified based on the World Health Organization criteria. Morphologically 241 of the samples were normal and the other 542 abnormal.</p><p><b>RESULTS</b>VCL, WOB, VAP of the morphologically abnormal group were significantly higher than those of the normal group (P < 0.05, P < 0.001), while MAD, LIN, STR of the abnormal group were significantly lower (P < 0.05, P < 0.001). There were significant positive correlations between the morphologically normal sperm rates and MAD, LIN, WOB, STR, and a significant negative correlation between the morphologically normal sperm rate and ALH.</p><p><b>CONCLUSION</b>Morphological abnormality of sperm is often accompanied with weak motility, which is probably attributed more to some factors that coact on both sperm motility and morphology than to the influence of sperm morphological abnormality on sperm motility.</p>

The effect of semen antisperm antibody on human sperm acrosin activity / 中华男科学杂志

<p><b>OBJECTIVES</b>To study the effect of antisperm antibody(AsAb) on human sperm acrosin activity.</p><p><b>METHODS</b>AsAb and sperm acrosin activity were measured and analyzed in 3,432 infertile men and 65 fertile volunteers.</p><p><b>RESULTS</b>AsAb positive rate was 10.20% in 3,432 case of male infertility, and 9.37% in 2,882 infertile males who received tests of sperm acrosin activity. Acrosin activity of infertility cases were lower than those of fertile cases(P < 0.001). The comparison between AsAb positive group and AsAb negative group infertility cases showed no significant differences of acrosin activity (P > 0.05). Between normal acrosin activity group and abnormal acrosin activity group, there was no significant difference of AsAb positive rate (P > 0.05).</p><p><b>CONCLUSIONS</b>Antisperm antibody could not affect acrosin activity.</p>