: ; (AH); ; (ET) : R711.6 : A : X(2014) (FET) ; 1985, Rall. ; 1990, Gordts. ,, (assisted hatching, AH),, Tyrode s.

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1 34 9 Vol.34 No Sep Reproduction & Contraception doi: /j.issn X randc_journal@163.com ( ) : : ( 2 ) 128 : (49.11% vs 48.28% 42.01% vs 42.36% 28.66% vs 28.35% P>0.05); (60.47% vs 63.16% 48.84% vs 55.26% 37.88% vs 38.57% P>0.05); (43.33% vs 45.59% 36.67% vs 39.71% 25.23% vs 26.15% P>0.05) (19.05% vs 4.76% P=0.153; 36.36% vs 11.11% P=0.035) (P<0.05) : : ; (AH); ; (ET) : R711.6 : A : X(2014) ; 1985 Rall [1] ; 1990 Gordts () [3] [4-6] [2] (assisted hatching AH) ( ) (ZZjdyx12106 ZZjdyx13098) : ; Tel/Fax: ; libinliccc@163.com [7] Tyrode s [8] [9] [10] [11] piezo [12-18] AH 718

2 1 1.1 [19-21] AH [2223] AH AH AH ~12 ; 3 min 3 ( 20% SSS 0.30 mol/l mhtf Origio) 4 ( 20% SSS mhtf Origio) 5 min 10%SSS (multiblast medium Irvine Scientific) 37 5%CO2 1.4 AH 1/4 70%~80% 37 5%CO (equilibration solution ES Origio)[ 20% (serum substitute supplement SSS); 7.5% (ethylene glycol EG); 7.5% (dimethyl sulphoxide DMSO) (modified human tubal fluid mhtf)] (vitrification solution VS Origio)( 20%SSS 15%EG 15% DMSO 0.58 mol/l mhtf) 45~60 s ES 80% ( 20%SSS 0.83 mol/l mhtf Origio) Strawtop 1 1 min 2 ( 20%SSS 0.42 mol/l mhtf Origio) 3 [24] 14 d hcg 14 d 1.6 (x s) SPSS20 t χ 2 P< (21~47) ( (1~21) (P>0.05)

3 1 Table 1 General information of patients (x s %) ; ( ) Age (year) BMI (kg/m 2 ) Times of previous transplant failures Causes of infertility Female Male Mix (142/212) (26/212) (44/212) (168/241) 7.88 (19/241) (54/241) Type of infertility (106/212) (112/241) Primary infertility (106/212) (129/241) Secondary infertility Insemination IVF ICSI IVF/ICSI (157/212) (52/212) 1.42 (3/212) (184/241) (52/241) 2.07 (5/241) Endometrial preparation (79/212) (80/241) Natural cycle (59/212) (76/241) Hormone replacement (74/212) (85/241) Ovulation (mm) Embryo survival rate (394/400) (458/469) (107/212) (122/241) (92/212) (107/241) (119/394) (28/92) (137/458) (30/107) (76/212) (16/92) (90/241) (17/107) (P>0.05) ; (P>0.05) (P<0.05) 3 2 Table 2 Effect of laser thinning on the cleavage stage frozen-thawed embryo transfer (x s %) ( ) Age (year) BMI (kg/m 2 ) (mm) (328/331) (388/396) Embryo survival rate (83/169) (98/203) (71/169) (86/203) (94/328) (110/388) (24/71) (24/86) (59/169) (70/203) (12/71) (16/86) 720

4 3 (x s %) Table 3 Effect of laser thinning on the frozen-thawed blastocyst transfer ( ) Age (year) BMI (kg/m 2 ) (mm) * (26/43) (24/38) (21/43) (25/66) (4/21) (17/43) (4/21) (21/38) (27/70) (6/21) (20/38) 4.76 (1/21) *: P<0.05 compared with the control Table 4 Effect of laser thinning on the frozen-thawed embryo transfer cycles of repeated implantation failure patients (x s %) ( ) Age (year) BMI (kg/m 2 ) (mm) Embryo survival rate (107/109) (130/133) (26/60) (31/68) (22/60) (27/107) (5/22) (14/60) (8/22) * (27/68) (34/130) (7/27) (24/68) (3/27) *: P<0.05 compared with the control ; IVF (P>0.05) [4-6] (P=0.035)

5 AH [2526] AH [27-29] AH AH ; Petersen Ng [2728] AH AH AH AH Germond [30] AH 1 AH Jun [20] AH AH [1] Rall WF Fahy GM. Ice-free cryopreservation of mouse embryos at 196 by vitrification. Nature (6003): [2] Gordts S Roziers P Campo R et al. Survival and pregnancy outcome after ultrarapid freezing of human embryos. Fertil Steril (3): [3] Rama Raju GA Haranath GB Krishna KM et al. Vitrification of human 8-cell embryos a modified protocol for better pregnancy rates. Reprod Biomed Online (4): [4] Manna C Rienzi L Greco E et al. Zona pellucida solubility and cortical granule complements in human oocytes following assisted reproductive techniques. Zygote (3): [5] Vanderzwalmen P Bertin G Debauche CH et al. Vitrification of human blastocysts with the hemi-straw carrier application of assisted hatching after thawing. Hum Reprod (7): [6] Cohen J Elsner C Kort H et al. Impairment of the hatching process following IVF in the human and improvement of implantation by assisting hatching using micromanipulation. Hum Reprod (1):7-13. [7] Cieslak J Ivakhnenko V Wolf G et al. Three-dimensiional partial zona dissection for preimplantation genetic diagnosis and assisted hatching. Fertil Steril (2): [8] Cohen J Alikani M Trowbridge J et al. Implantation enhancement by selective assisted hatching using zona drilling of human embryos with poor prognosis. Hum Reprod (5): [9] (3): [10] Nakayama T Fujiwara H Tastumi K et al. A new assisted hatching technique using a piezo-micromanipulator. Fertil Steril (4): [11] Montag M van der Ven H. Laser-assisted hatching in assisted reproduction. Croat Med J (3): [12] Takahashi K Takenaka M Ishizuka B. The effect of assisted hatching on patients repeatedly failed to conceive by in vitro fertilization. Nippon Sanka Fujinka Gakkai Zasshi (10): [13] (2):

6 [14] Edi-Osagie E Hooper L Seif MW. The impact of assisted hatching on live birth rates and outcomes of assisted conception: a systematic review. Hum Reprod (9): [15] Blessmann-Roset J Rives N Clavier B et al. Laser assisted hatching: rouen university hospital outcomes. Gynecol Obstet Fertil (4): [16] (10): [17] Parikh FR Kamat SA Nadkarni S et al. Assisted hatching in an in vitro fertilization programme. J Reprod Fertil Suppl : [18] (9): [19] Bider D Livshits A Yonish M et al. Assisted hatching by zona drilling of human embryos in women of advanced age. Hum Reprod (2): [20] Edirisinghe WR Ahnonkitpanit V Promviengchai S et al. A study failing to determine significant benefits from assisted hatching: patients selected for advanced age zonal thickness of embryos and previous failed attempts. J Assist Reprod Genet (6): [21] Petersen CG Mauri AL Baruffi RL et al. Laser-assisted hatching of cryopreserved-thawed embryos by thinning on quarter of the zona. Reprod Biomed Online (5): [22] Schieve LA Meikle SF Peterson HB et al. Does assisted hatching pose a risk for monozygotic twinning in pregnancies conceived through in vitro fertilization? Fertil Steril (2): [23] Jun SH Milki AA. Assisted hatching is associated with a higher ectopic pregnancy rate. Fertil Steril (6): [24]. GnRH-a hmg (6): [25] de Moraes LA de Miranda Cota AM. Assisted hatching improves implantation rates on cryopreserved-thawed embryos. A randomized prospective study. Hum Reprod (5):1429. [26] Balaban B Urman B Yakin K et al. Laser-assisted hatching increases pregnancy and implantation rates in cryopreserved embryos that were allowed to cleave in vitro after thawing: a prospective randomized study. Hum Reprod (8): [27] Ng EH Naveed F Lau EY et al. A randomized double-bline controlled study of the efficacy of laser-assisted hatching on implantation and pregnancy rates of frozed-thawed embryo transfer at the cleavage stage. Hum Reprod (4): [28] Petersen CG Mauri AL Baruffi RL et al. Laser-assisted hatching of cryopreserved-thawed embryos by thinning on quarter of the zona. Reprod Biomed Online (5): [29] Edirisinghe WR Ahnonkitpanit V Promviengchai S et al. A study failing to determine significant benefits from assisted hatching: patients selected for advanced age zonal thickness of embryos and previous failed attempts. J Assist Reprod Genet (6): [30] Germond M Nocera D Senn A et al. Microdissection of mouse and human zona pellucida using a 1.48-microns diode laser beam: efficacy and safety of the procedure. Fertil Steril (3): ( ) Effect of Laser Thinning Assisted Hatching on Vitrified-warmed Embryo Transfer Zheng YAN Yan-ping KUANG Song-guo XUE Qi-feng LV Bin LI (Reproductive Medicine Department of Shanghai Ninth People s Hospital Affiliated Shanghai Jiao Tong University School of Medicine Shanghai ) ABSTRACT Objective: To explore the effect of laser thinning assisted hatching on frozen-thawed embryo transfer outcome. Methods: Selected embryos from 372 cleavage stage embryo transfer cycles 81 blastocyst transfer cycles and 128 repeated implantation failure cycles were processed and divided into laser thinning group and control group respectively according to odd-numbered days and even-numbered days of embryo thawed day. 723

7 The embryos in laser thinning group received laser thinning assisted hatching (AH) treatment before fertilization and which in control group did not. The laboratory and clinical outcomes were compared between the two groups. Results: The biochemical pregnancy rate the clinical pregnancy rate and the implantation rate in the assisted hatching cleavage stage embryo group showed no significant differences with those of the control (49.11% vs 48.28% 42.01% vs 42.36% 28.66% vs 28.35% P>0.05); There was no statistical difference of biochemical pregnancy rate clinical pregnancy rate and implantation rate between the assisted hatching blastocyst group and the control group (60.47% vs 63.16% 48.84% vs 55.26% 37.88% vs 38.57% P>0.05); the application of laser assisted hatching did not improve the pregnancy outcomes of repeated implantation failure patients (43.33% vs 45.59% 36.67% vs 39.71% 25.23% vs 26.15% P>0.05). However in the blastocyst transfer patients and repeated implantation failure patients there was an increasing trend of abortion rate in laser assisted hatching group (19.05% vs 4.76% P=0.153; 36.36% vs 11.11% P=0.035). Conclusion: The laser assisted hatching technique does not improve pregnancy outcomes of frozen-thawed embryo transfer patients and its long-term safety needs further study. The clinical application of this method should be cautious. Key words: laser; assisted hatching (AH); vitrification freezing; embryo transfer (ET) &&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&& ( 702 refer to p702) A Dynamic Method of Embryo Culture Based on A Microfluidic Chip Wei WANG Ya-ya PENG Guang-tie LIANG Zao-wen LIAO Da-yu LIU (Department of Laboratory Medicine Guangzhou First People s Hospital Guangzhou Medical University Guangzhou ) ABSTRACT Objective: To develop a microfluidic chip for embryo dynamic culture method nearly physiological status. Methods: A microfluidic chip was designed and fabricated for testing the influence of continual fluid flow on embryo development. This gave rise to design of a microfluidics system using microchannels as conduits for fluid flow through a 16-microcellular where mouse embryos resided by mimicking the fluidmechanical and biochemical stimulation embryos experience in vivo from ciliary currents and oviductal contractions. And compared with conventional droplet culture method monitoring of the embryonic development conditions and blastocyst formation rate. Results: The microfluidic chip dynamic culture method can significantly improve the embryo development. The microfluidic dynamic method was superior to the microdrop-static method in terms of 4-cell embryo rate ( % vs %) morula rate ( % vs %) and blastocyst rate ( % vs %)(P<0.05). These two methods didn t show significant difference in the rate of 2-cell embryos ( % vs % P>0.05). Conclusion: Physiological embryo culture was achieved on a microfluidic chip. This microfluidic method was able to improve embryo development and showed advantage over the conventional method which was expected to serve as a powerful tool for embryo culture in the future. Key words: microfluidic chip; dynamic culture; embryo development 724

2013 Sep.; 24(3):

Journal of Reproduction & Contraception doi: 10.7669/j.issn.1001-7844.2013.03.0151 2013 Sep.; 24(3):151-158 E-mail: randc_journal@163.com Reducing the Trigger Dose of Human Chorionic Gonadotrophin Does