Comparison of the Developmental Potential and Clinical Results of In Vivo Matured Oocytes Cryopreserved with Different Vitrification Media / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Oocyte vitrification is widely used throughout the world, but its clinical efficacy is inconsistent and depends on the vitrification media. This study compared the developmental potential and clinical results of in vivo matured oocytes cryopreserved with different vitrification media.</p><p><b>METHODS</b>This retrospective study involved vitrified-warmed oocytes at one in vitro fertilization laboratory. Vitrification media kits comprised the MC kit (ethylene glycol [EG] plus 1,2-propanediol [PROH]), the KT kit (EG plus dimethyl sulphoxide [DMSO]), and the Modified kit (EG plus DMSO and PROH kit). Rates of oocyte survival and subsequent developmental potential were recorded and analyzed. The t-test and the Chi-square test were used to evaluate each method's efficacy.</p><p><b>RESULTS</b>Oocyte survival rate was significantly higher for the Modified kit (92.0%) than for the MC kit (88.2%) (P < 0.05) and the KT kit (77.3%) (P < 0.001). The rate of high-quality embryo development in the Modified kit group (35.8%) was significantly higher than in the MC kit group (29.0%) and the KT kit group (28.3%) (P < 0.001). No significant differences were observed in the clinical pregnancy and implantation rates among the MC, KT, and Modified kit groups (37.2% vs. 30.2% vs. 39.6%; 21.9% vs. 18.8% vs. 27.4%, respectively) (P > 0.05). The high-quality embryo rate per warmed oocyte was significantly higher (23.4%) in the Modified kit group than in the other groups (P < 0.001). The embryo utilization and live birth rates per warmed oocyte were the highest in the Modified kit group, but not significantly (P > 0.05).</p><p><b>CONCLUSIONS</b>Modified vitrification media are efficient for oocyte vitrification and, with further verification, may be able to replace commercially available media in future clinical applications.</p>

Application of calcium ionophore A23187 in ICSI for globozoospermia: A report of 2 cases and review of the literature / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the pathogenesis of globozoospermia, fertilization ability of round-headed sperm, and the application value of assisted oocyte activation in intracytoplasmic sperm injection (ICSI) for the wives of glohozoospermia men.</p><p><b>METHODS</b>We collected oocytes from the wives of 2 globozoospermia patients and randomly divided them into two groups after ICSI to receive calcium ionophore A23187-activation and conventional treatment, respectively. We reviewed the relevant literature published at home and abroad, and discussed the etiology of globozoospermia, fertilization ability of round-headed sperm, and treatment options for this disease.</p><p><b>RESULTS</b>Quality embryos were obtained in the A23187-activation group while no fertilized oocytes, oocyte cleavage, quality embryos, or blastular formation were found in the conventional treatment group. Both women achieved pregnancy and gave birth to healthy neonates after transfer of the quality embryos from the A23187-activation group.</p><p><b>CONCLUSION</b>Calcium ionophore A23187 can be applied to ICSI for the wives of globozoospermia men and bring about desirable clinical outcomes. Meanwhile, attention should be paid to its safety.</p>

Comparison of the outcome of intracytoplasmic single sperm injection between two motile sperm separation methods / 中华男科学杂志

<p><b>OBJECTIVE</b>To evaluate the efficiency of density gradient and improved swim-up methods for motile sperm isolation from fresh semen samples in intracytoplasmic sperm injection and embryo transfer (ICSI-ET) program, thus guiding the clinical application.</p><p><b>METHODS</b>The fertilization rate, cleavage rate, high-quality embryo rate, clinical pregnancy rate, sperm abnormal rate and recovery rate of 42 cycles were studied prospectively. The cycles were divided into two groups with respect to the motile sperm isolation methods.</p><p><b>RESULTS</b>No obvious difference was found in the fertilization rate, cleavage rate, high-quality embryo rate and clinical pregnancy rate between the two methods. The abnormal sperm rate induced by the improved swim-up method was significantly higher than that of the density gradient method (P < 0.01). For severe oligozoospermia, the recovery rate of motile sperm with density gradient was higher than that of improved swim-up (P < 0.01).</p><p><b>CONCLUSION</b>In the ICSI-ET, there is no obvious difference in clinical pregnancy outcome between the two methods. The method of improved swim-up can be used for all patients but those with severe oligozoospermia.</p>

Sex chromosome analysis and IGF-II expression on activated human unfertilized oocytes after ICSI with calcium ionophore A23187 and puromycin / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the sex chromosomes and the expression of insulin-like growth factor-II (IGF-II) on activated human unfertilized oocytes after intracytoplasmic sperm injection(ICSI) with calcium ionophore A23187 and puromycin.</p><p><b>METHODS</b>All 95 discarded oocyes that showed no evidence of fertilization at 16-18 h after in vitro maturation and intracytoplasmic sperm injection cycles (IVM-ICSI)/conventional ICSI were exposed to calcium ionophore A23187 (5 micromol/L) for 5 min and then were incubated with puromycin (10 microg/mL) for 4 h. After activation, the oocytes were cultured in vitro for 3-5 days. The sex chromosome analysis was performed by dual color fluorescence in situ hybridization. The expression of IGF-II on the activated embryos, normal embryos, and parthenotes was examined.</p><p><b>RESULTS</b>The combination of calcium ionophore A23187 with puromycin could activate the unfertilized oocytes 22 h after ICSI. The activated rate, cleavage rate, and quality of activated embryos of the IVM-ICSI group were similar to those of ICSI group, respectively. Sex chromosome analysis indicated that 8 male and 5 female embryos had been derived from two pronucleus and a second polar body. The expression of IGF-II on activated embryos and normal embryos was high and similar, which was much stronger than that of parthenotes.</p><p><b>CONCLUSION</b>The combination of calcium ionophore A23187 with puromycin could effectively activate unfertilized oocytes 22 h after ICSI. Moreover, the unfertilized oocytes activated by calcium ionophore A23187 and puromycin had normal sex chromosomes and expression of IGF-II like the normal embryos. These suggest that oocyte activation may be considered as a remedial measure in the presence of total or nearly total fertilization failure in ICSI.</p>

Comparison of vitrification and slow-freezing of human day 3 cleavage stage embryos:postvitrification development and pregnancy outcomes / 中华妇产科杂志

Objective To compare the effects of vitrification with slow-freezing on the developmental ability of day 3 cleavage stage embryos.Methods Patients who had no less than 4 high quality embryos were included in this study.These embryos were cryopreserved using the methods of vitrification or slow-freezing.In the eryopreserved embryo transfer cycles,the embryos which were cryopreserved using one of the methods were chosen randomly.The developmental ability of embryos was compared between these two groups.Results A total of 80 patients were included in this study with 160 embryos.In the group of slow-freezing,73(91%)embryos were survived and achieved 15(38%)clinical pregnancies.Among these,3 were twins and the implantation rate was 25%(18/73).In the group of vitrification,71(89%)embryos were survived and achieved 19(48%)clinical pregnancies.Among these, 9 were twins and the implantation rate was 39%(28/71),which was significantly higher than the slow- freezing group(P

Spindle and chromosome configuration of human in vitro matured oocytes after slow freezing-fast thawing / 中华妇产科杂志

Objective To evaluate the development of immature oocytes after freezing-thawing by conventional cryopreservation method for mature oocytes.Methods Immature oocytes were collected from stimulated ovaries of intracytoplasmic sperm injection(ICSI)cycles.Immature oocytes were in vitro matured directly or after slow freezing-fast thawing and immunostained for tubulin and chromatin and at last visualized by confocal microscopy.Results No statistical difference was found in maturity rate between freezing groups and the controls.There was a statistically significant increase in abnormalities of chromosome(23.7% vs. 50%)and spindle(28.9% vs.53.9%)in the GV freezing group compared with the GV control(P