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.

Effects of different doses of letrozole on the incidence of early-onset ovarian hyperstimulation syndrome after oocyte retrieval.

We explored the effects of different doses of letrozole on the incidence of ovarian hyperstimulation syndrome (OHSS) after oocyte retrieval during in vitro fertilization (IVF) in patients with high-risk OHSS. A total of 88 patients were randomly divided into a control group, and groups treated with 2.5 mg, 5 mg, or 7.5 mg of letrozole. We found that from the fifth day after human chorionic gonadotrophin (hCG) treatment, the E2 level decreased and there were statistical differences between the four groups (p < 0.05). From the eighth day after hCG treatment, the luteinizing hormone (LH) level increased, but the progesterone (P) level decreased. There were statistical differences between groups (p < 0.05). From the fifth day after hCG treatment, the level of vascular endothelial growth factor (VEGF) increased in the control, but decreased in the letrozole groups in a dose-dependent manner. There were statistically significant differences between groups (p < 0.001). The incidence of moderate and severe OHSS was lower in the 7.5 mg group than in the control group (p < 0.05). In the patients with high-risk OHSS undergoing whole embryo frozen transfer, treatment with 7.5 mg letrozole may be useful to limit OHSS.

[Relationship between letrozole administration during the luteal phase after oocyte retrieval and the early-stage ovarian hyperstimulation syndrome oocurrence].

OBJECTIVE: To investigate the effect of letrozole in decreasing the early-stage ovarian hyperstimulation syndrome (OHSS) occurrence during the luteal phase for patients of OHSS high-risk after oocyte retrieval. METHODS: A total of 176 high-risk OHSS patients were randomly divided into two groups after oocyte retrieval. Patients in experiment group (n = 86) received 5 mg letrozole per day from the retrieval day and last for 5 days. Others in control group (n = 90) received placebo. The serum concentration of FSH, LH, estradiol (E2), progesterone (P) and vascular endothelial growth factor (VEGF) from the day of hCG injection to days after injection (5 days, 8 days, 10 days) were measured. And the incidence of moderate and severe OHSS was observed. RESULTS: The concentration of E2 on the indicated days (5 days, 8 days, 10 days after hCG injection) in experiment group and control group were (5 727±2 089) versus (11 826±4 281) pmol/L, (1 613±879) versus (7 925±3 507) pmol/L, (193± 90) versus (1 628±888) pmol/L; the concentration of VEGF on the indicated days in the two groups were (80±14) versus (108±19) ng/L, (66±11) versus (126±14) ng/L, (48±7) versus (148±14) ng/L; the concentration of E2 and VEGF were lower than those in control group (all P < 0.01). The FSH concentration in experiment group were (2.1±1.1) and (3.5±1.3) U/L on the day of fifth and eighth day after hCG injection, which were significantly higher than (0.7±0.3) and (0.7±0.4) U/L in control group (P < 0.05); the LH concentration in experiment group were (0.26±0.19) and (0.72±0.60) U/L on the day of fifth and eighth day after hCG injection, which were significantly higher than (0.11±0.03) and (0.14±0.08) U/L in control group (P < 0.05). The incidence of moderate and severe OHSS was signicantly decreased after letrozole treatment compared with control group [2% (2/86) versus 12% (11/90), P < 0.05]. CONCLUSION: Administration of 5 mg/d letrozole for 5 days during the luteal phase can reduce the E2 and VEGF levels for the high-risk OHSS patients who needed cryopreserve all embryos, and also reduce the occurrence of early OHSS.

[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.

Phospholipid langmuir film as template for in situ silica nanoparticle formation at the air/water interface.

This study describes for the first time the in situ formation of small-sized silica nanoparticles after the polycondensation of tetraethylorthosilicate (TEOS) at the air/water interface. Mixtures of TEOS and 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) are prepared with different molar ratios (TEOS/DPPC 15:1, 50:1, 500:1, and 5000:1) and dissolved in chloroform. Spreading TEOS/DPPC 15:1 and 50:1 on the water surface of a Langmuir trough leads to an initial increase of the surface pressure ( approximately 26-29 mN/m) after allowing chloroform to evaporate. Within the reaction time of 21 h, only a slight decrease of the surface pressure by approximately 2 mN/m occurs. Films of silica/DPPC mixtures are transferred from the air/water interface to silicon wafers by dip-coating. The morphologies of the silica nanoparticles and agglomerates together with DPPC are observed using tapping mode atomic force microscopy (AFM). An ordered array of silica nanoparticles can be observed after 21 h reaction of the precursor solution of TEOS/DPPC 500:1 and transfer onto hydrophilic silicon substrate. A mixture with a molar ratio of TEOS/DPPC of 5000:1 is also placed onto the water surface. DPPC forms a uniform film observed by AFM after film transfer. Silica rich domains can be observed with mesoporous morphology.

Hierarchical organization of poly(ethylene oxide)-block-poly(isobutylene) and hydrophobically modified Fe(2)O(3) nanoparticles at the air/water interface and on solid supports.

Langmuir monolayers and Langmuir-Blodgett (LB) film morphologies of block copolymers and hydrophobically modified iron oxide nanoparticles were studied by surface pressure-mean molecular area (pi-mmA) measurements and by tapping mode atomic force microscopy (AFM). The amphiphilic diblock copolymers consisted of a hydrophilic poly(ethylene oxide) (PEO) block and a hydrophobic poly(isobutylene) (PIB) block. The pi-mmA isotherm of PEO(97)-b-PIB(37) (the subscripts refer to the respective degrees of polymerization) at the air/water interface had an extended plateau reflecting the extension of PEO chains into the water subphase at a surface pressure of 10 mN.m(-1), which is absent for the more hydrophobic PEO(19)-b-PIB(130). Iron oxide (Fe(2)O(3)) nanoparticles capped with oleic acid ligands as the shell were dispersed in the amphiphilic block copolymers at the air/water interface to prevent macroscopic aggregation of the particles. When the nanoparticles were mixed with PEO(97)-b-PIB(37), using a particle to polymer chain ratio of 1:100, macroscopic aggregation of the nanoparticles was not observed, and the pi-mmA isotherm was dominated by PEO(97)-b-PIB(37). Monolayers of block copolymers were transferred at different surface pressures from the air/water interface to hydrophilic silicon substrates using the Langmuir-Blodgett technique. The AFM images of PEO(97)-b-PIB(37) LB films depicted not only the typical finger-like morphology of the crystallized PEO blocks but also PIB blocks arranged in vertical columns growing perpendicular to the substrate surface. The columns are characteristic for PEO(19)-b-PIB(130) LB films after transfer at high surface pressures and can be assigned to a mesomorphic PIB phase with ordered chains. Finally, it was observed that small clusters of a few Fe(2)O(3) nanoparticles occupy the top of PIB phases after compression and transfer of the block copolymer nanoparticle mixtures to solid supports.

[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.