Objective To investigate the effects on pregnancy outcome and neonate by artificial shrinkage by microsucting the fluid of expanded blastocysts before vitrification using glass micropipette (GMP).Methods From Jan.2006 to Dec.2009, 342 vitrified-thawed blastocyst cycles from patients that performed in vitro fertilization-embryo transfer (IVF-ET) or intracyteplasmic sperm injection ( ICSI ) were enrolled in this study in Reproductive Medicine center, Maternal and Child Health Hospital of Guangxi Zhuang Autonomous Region.Three hundred and fourteen cycles of expanded blastocysts were artificially shranked by microsucting blastocoelic fluid with a micro-needle before vitrification as artificial shrinkage group, in the mean time, 28 cycles without artificial shrinkage were chosed as control group.The survival rate, implantation rate, clinical pregnancy rate and transfer canceled rate were compared between artificial shrinkage group and control group.Among pregnant women, the miscarriage rate, live birth rate, congenital birth defect rate, neonatal weight and gestational age were compared with those of fresh embryo transfers in 520 cycles.Results The blastocyst survival rate, implantation rate and clinical pregnancy rate were 95.3%(403/423), 38.0% ( 153/403), 44.6% (140/314) in artificial shrinkage group and 64.3 % (27/42),7.4% (2/27), 7.1% (2/28) in control group, respectively, which reached statistical difference (P＜0.05).The transfer canceled rate was 0 in artificial shrinkage group and 25.0% (7/28) in control group, which also reached statistical difference ( P ＜ 0.05 ).Among pregnant patients, the miscarriage rate of 18.2% (10/55), live birth rate of 80.0% (44/55), gestational age of (38.2 ± 1.3) weeks, congenital birth defect rate of 2.1% (1/47), birth weight of newborns of (2989 ±640) gram in artificial shrinkage group were not significantly different with 17.5% (91/520), 74.0% (385/520), (37.9 ±2.3) weeks,1.7% (8/479) and (2856±640) gramin fresh embryo transfer group (P＞0.05).Conclusion Artificial shrinkage by microsucting blastocoelic fluid with a micro-needle before vitrification significantly improved the vitrification effects of expanded blastocyst and no distinct increasing rate of neonates congenital anomality were observed.

Objective To investigate the effect of blastocyst quality on the strategy of single blastocyst transfer in frozen-thawed cycles. Methods A retrospective analysis was performed in Reproductive Medicine Center, Maternal and Child Health Hospital of Guangxi Zhuang Autonomous Region on clinical data of single frozen-thawed blastocyst transfer cycles from January 2008 to December 2013. All cycles were divided into four groups (AA, AB/BA, BB, BC/CB) according to the blastocyst score, then the clinical outcomes were compared between groups. And on this basis, the clinical outcomes were further explored when the group of outcomes with single blastocyst transfer wasn′t ideal, which would diverted to transfer two blastocyst. Results In single frozen blastocyst transfer cycles, the clinical pregnancy rate of each group with the blastocyst scored AA, AB/BA, BB, BC/CB were 61.4%(470/765), 51.2%(330/645), 40.5%(407/1 005), 22.9%(60/262), live births rate in each group were 52.2%(399/765), 41.2%(266/645), 30.4%(306/1 005), 13.7%(36/262), and the abortion rate were 13.6%(64/470), 16.7%(55/330), 21.4%(87/407), 35.0%(21/60), separately. This showed that the clinical pregnancy rate and live births rate decreased significantly with the decline of blastocyst quality (P<0.01), but the abortion rate showed significant upward trend (P<0.01). When single blastocyst scored≥BB grade transferred, an acceptable clinical pregnancy rate (>40%) and live births rate (>30%) could be obtained, however, the clinical pregnancy rate of 22.9% and live births rate of 13.7%could only be acquired when blastocyst scored BC/CB only transferred one embryo, which significant lower than those of each group scored ≥BB grade (P<0.01). So, after that, the blastocyst scored BC/CB were further divided into two groups (single blastocyst transferred versus two blastocyst transferred) to investigate, then the result showed that the clinical pregnancy rate [22.9%versus 38.5%(67/174), P<0.01] and live births rate [13.7%versus 30.5%(16/67), P<0.01] were significantly increased in the group of two blastocyst transferred compared with the group of one blastocyst transferred, and the abortion rate was also significantly decreased from 35.0%to 17.9%(12/67;P<0.05). So when two blastocyst scored BC/CB were transferred, the clinical outcomes were similar to the group of one blastocyst scored BB transferred (P>0.05). Conclusions Of single blastocyst transfer in frozen-thawed cycles, the clinical pregnancy rate and liver births rate showed significant upward trend, but the abortion rate showed significant downward trend, with the decline of blastocyst quality. When the blastocyst scored ≥BB grade, the single blastocyst transfer could be considered to be performed.

The aim of this study was to report a successful pregnancy using completely immotile frozen-thawed spermatozoa selected by laser. A single laser shot was used to detect the presence of viable immotile spermatozoa in fresh and frozen-thawed testicular spermatozoa. The viability rate was 55.8% after the laser detection, and cryopreservation was carried out immediately. The thawing test was performed on the day of oocyte pick-up, and no motile sperm were observed after extending the culture for another 4 hours, while a survival rate of 39.8% was detected using the laser. In all, five mature oocytes were injected, resulting in four cases of normal fertilization (80%) on day 1. Further, two high-quality day 3 embryos were transferred, which resulted in a singleton pregnancy. Our study demonstrates that completely immotile spermatozoa are worth cryopreserving for further intracytoplasmic sperm injection, which provides a new insight into male fertility preservation in cases of completely immotile spermatozoa.
Cryopreservation ; Embryonic Structures ; Fertility Preservation ; Fertilization ; Humans ; Male ; Oocytes ; Pregnancy* ; Sperm Injections, Intracytoplasmic ; Spermatozoa* ; Survival Rate

A few of flow experiments were utilized to verify a theoretical hypothesis proposed by Fung and coworker which showed that the tensile stress in the upper cell membrane of the vascular endothelium could accumulate upstream against the direction of blood flow. Endothelial cells from replicate human umbilical vein segment (HUVSEC) in vitro with length of 11 cm and 21 cm were exposed to the same pulsatile laminar shear stress averaged of 0.12 N/m2 for 42 hours. The average production rate of endothelin-1(ET-1), at 11 cm segment is 50% lower than that at 21 cm segment(16.93 +/- 0.89) vs. (26.13 +/- 1.79) pg/cm2.h respectively. The average production rate of ET-1 under pulsatile laminar flow was significantly higher than that under steady laminar flow. It showed that, high correlation of the length of HUVSEC with their ET-1 metabolism exists, suggesting that the tensile stress in the upper endothelial cell membrane could accumulate.
Cell Membrane ; physiology ; Endothelin-1 ; metabolism ; Endothelium, Vascular ; cytology ; physiology ; Humans ; In Vitro Techniques ; Infant, Newborn ; Stress, Mechanical ; Umbilical Veins ; anatomy & histology

OBJECTIVE: The aim of this study was to explore the effects of the insemination method on the outcomes of elective blastocyst culture. METHODS: We retrospectively analyzed the outcomes of elective blastocyst culture performed between January 2011 and December 2014. RESULTS: There were 2,003 cycles of conventional in vitro fertilization (IVF) and 336 cycles of intracytoplasmic sperm injection (ICSI), including 25,652 and 4,164 embryos that underwent sequential blastocyst culture, respectively. No significant differences were found in the female patients' age, basal follicle-stimulating hormone level, basal luteinizing hormone level, body mass index, number of oocytes, maturity rate, fertilization rate, or good-quality embryo rate. However, the blastocyst formation rate and embryo utilization rate were significantly higher in the conventional IVF group than in the ICSI group (54.70% vs. 50.94% and 51.09% vs. 47.65%, respectively, p<0.05). The implantation/pregnancy rate (IVF, 50.93%; ICSI, 55.10%), miscarriage rate (IVF, 12.57%; ICSI, 16.29%), and live birth rate (IVF, 42.12%; ICSI, 44.08%) were similar (p>0.05). No cycles were canceled due to the formation of no usable blastocysts. CONCLUSION: Although the fertilization method had no effect on clinical outcomes, the blastocyst formation rate and embryo utilization rate in the ICSI group were significantly lower than those observed in the conventional IVF group. Therefore, more care should be taken when choosing to perform blastocyst culture in ICSI patients.
Abortion, Spontaneous ; Blastocyst* ; Body Mass Index ; Embryonic Structures ; Female ; Fertilization ; Fertilization in Vitro ; Follicle Stimulating Hormone ; Humans ; Insemination* ; Live Birth ; Luteinizing Hormone ; Methods* ; Oocytes ; Pregnancy ; Pregnancy Rate ; Retrospective Studies ; Sperm Injections, Intracytoplasmic