Clinical outcomes of frozen-thawed single blastocyst transfer in patients requiring whole embryo freezing.

We compared clinical outcomes amongst frozen-thawed cleavage-stage embryo, double and single blastocyst transfers in patients requiring whole embryo freezing. Data of infertile patients undergoing in-vitro fertilization and embryo transfer (IVF-ET) in our Reproductive Medicine Center from January 2010 to December 2012 were retrospectively analyzed. According to patients' wishes, patients were divided into cleavage-stage embryo transfer groups (group A, n = 456), double blastocyst transfer group (group B, n = 106), and single blastocyst transfer group (group C, n = 402). We found that the number of frozen embryos was significantly less in groups B and C than in group A (all p < 0.05), but the implantation rate was significantly higher in groups B and C as compared to group A (all p < 0.05). The clinical pregnancy rate and pregnancy rate per included cycle were significantly higher in group B than in groups A and C (all p < 0.05). The multiple pregnancy rate was significantly lower in group C than in groups A and B (all p < 0.05). The rate of early abortion was significantly lower in group C as compared to group A (p < 0.05). The data support the view that it may be the best clinical strategy for patients who require whole embryo freezing and have four or more Day 3 embryos available, to incubate Day 3 embryos into blastocysts, which are then vitrified for elective single blastocyst transfer.

[Study on ovary development in vitrification of embryos born mice and expression of growth differentiation factor 9].

OBJECTIVE: To study ovarian development in vitrification of embryos born mice and expression of growth differentiation factor 9 (GDF-9) in its. METHODS: The vitrification recovery embryos (vitrified-embryo group) and fresh embryos (fresh-embryo group) were transplanted into pseudopregnant mice, respectively. The female offspring mice in two groups were sacrificed on the 3(rd), 7(th), 14(th), 21(st), 28(th) and 60(th) day after birth, the ovarian tissues were taken, 6 mice in each time point of each group. The ovarian development was observed by HE staining, the expression of GDF-9 mRNA and protein at each time point of two groups were detected by reverse transcription (RT)-PCR and western blot. RESULTS: HE staining showed that no abnormal ovarian development was observed in offsprings at two groups. On the 3(rd), 7(th), 14(th), 21(st), 28(th) and 60(th) day after birth, the expression of GDF-9 mRNA in vitrified-embryo group were 0.14 ± 0.07, 0.42 ± 0.16, 1.00 ± 0.24, 1.59 ± 0.28, 2.05 ± 0.32 and 2.23 ± 0.21, respectively, which in fresh-embryo group were 0.13 ± 0.06, 0.45 ± 0.18, 1.00 ± 0.21, 1.56 ± 0.26, 2.01 ± 0.37 and 2.26 ± 0.23, respectively, there was no statistical difference between two groups (P > 0.05); the expression of GDF-9 protein in vitrified-embryo group were 0.040 ± 0.030, 0.120 ± 0.060, 0.170 ± 0.030, 0.250 ± 0.040, 0.320 ± 0.060 and 0.330 ± 0.010, respectively, which in fresh-embryo group were 0.030 ± 0.020, 0.110 ± 0.040, 0.150 ± 0.010, 0.210 ± 0.020, 0.360 ± 0.070 and 0.350 ± 0.030, respectively, there was no statistical difference between two groups (P > 0.05). CONCLUSION: The ovarian morphology in vitrification of embryos born mice and expression of GDF-9 in ovary has no any obvious change.

[Outcomes of ICSI with sperm from different sources: a retrospective study of 431 cycles].

OBJECTIVE: To compare the outcomes of intracytoplasmic sperm injection (ICSI) with retrieved epididymal and testicular sperm for obstructive azoospermia and with ejaculated sperm for severe oligozoospermia and asthenospermia. METHODS: We retrospectively analyzed 431 ICSI cycles, which were divided according to sperm sources into Groups A (n=287 in patients with severe oligozoospermia or asthenospermia using ejaculated sperm), B (n=109 in obstructive azoospermia patients with sperm retrieved by percutaneous epididymal sperm aspiration, PESA) and C (n=35 in obstructive azoospermia patients with sperm retrieved by testicular sperm extraction, TESE). Comparisons were made among the three groups in the rates of embryo implantation, fertilization, pregnancy, cleavage, and miscarriage. RESULTS: Group A showed statistically significant differences from Groups B and C in the rates of embryo implantation and pregnancy (18.46% vs. 25.23% and 28.76%, 31.23% vs. 42.16% and 39.39%, P < 0.05). But no significant differences were seen in the rates of fertilization, cleavage and miscarriage among the three groups (P > 0.05). CONCLUSION: The rates of embryo implantation and clinical pregnancy are higher in patients with obstructive azoospermia than in those with severe oligozoospermia or asthenospermia after ICSI with ejaculated sperm.

Construction and analysis of SSH cDNA library of human vascular endothelial cells related to gastrocarcinoma.

AIM: To construct subtracted cDNA libraries of human vascular endothelial cells (VECs) related to gastrocarcinoma using suppression substractive hybridization (SSH) and to analyze cDNA libraries of gastrocarcinoma and VECs in Cancer Gene Anatomy Project (CGAP) database. METHODS: Human VECs related to gastric adenocarcinoma and corresponding normal tissue were separated by magnetic beads coupled with antibody CD31 (Dynabeads CD31). A few amount of total RNA were synthesized and amplified by SMART PCR cDNA Synthesis Kit. Then, using SSH and T/A cloning techniques, cDNA fragments of differentially expressed genes in human VECs of gastric adenocarcinoma were inserted into JM109 bacteria. One hundred positive bacteria clones were randomly picked and identified by colony PCR method. To analyze cDNA libraries of gastrocarcinoma and VECs in CGAP database, the tools of Library Finder, cDNA xProfiler, Digital GENE Expression Displayer (DGED), and Digital Differential Display (DDD) were used. RESULTS: Forward and reverse subtraction cDNA libraries of human VECs related to gastrocarcinoma were constructed successfully with SSH and T/A cloning techniques. Analysis of CGAP database indicated that no appropriate library of VECs related to carcinoma was constructed. CONCLUSION: Construction of subtraction cDNA libraries of human VECs related to gastrocarcinoma was successful and necessary, which laid a foundation for screening and cloning new and specific genes of VECs related to gastrocarcinoma.