但COH劑量仍較一般病患劑量為高 (4206 vs 3961 IU).
J Assist Reprod Genet. 2016 Mar 4. [Epub ahead of print]
2016年4月30日
MTX並不明顯對卵巢產生永久性傷害(活產率35%)
但COH劑量仍較一般病患劑量為高 (4206 vs 3961 IU).
J Assist Reprod Genet. 2016 Mar 4. [Epub ahead of print]
The purpose of this research was to study whether methotrexate (MTX) as treatment for ectopic pregnancy (EP) impacts the future fertility of women undergoing assisted reproductive technology (ART) METHODS: In a systematic review and multi-center retrospective cohort from four academic and private fertility centers, 214 women underwent an ART cycle before and after receiving MTX as treatment for an EP. Measures of ovarian reserve and responsiveness and rates of clinical pregnancy (CP) and live birth (LB) were compared in the ART cycles prior and subsequent to MTX.
Seven studies were identified in the systematic review, and primary data from four institutions was included in the final analysis. Women were significantly older in post-MTX cycles (35.3 vs 34.7 years). There were no differences in follicle stimulating hormone, antral follicle count, duration of stimulation, oocytes retrieved, or fertilization rate between pre- and post-MTX cycles. However, post-MTX cycles received a significantly higher total dose of gonadotropins (4206 vs 3961 IU). Overall, 42 % of women achieved a CP and 35 % achieved a LB in the post-MTX ART cycle, which is similar to national statistics. Although no factors were identified that were predictive of LB in young women, the number of oocytes retrieved in the previous ART cycle and current AFC were predictive of LB (AUC 0.76, 0.75) for the older women.
MTX does not influence ovarian reserve, response to gonadotropin stimulation, and CP or LB rate after ART. MTX remains a safe and effective treatment option for women with asymptomatic EPs.
但COH劑量仍較一般病患劑量為高 (4206 vs 3961 IU).
J Assist Reprod Genet. 2016 Mar 4. [Epub ahead of print]
輕劑量賀爾蒙補充可達到最佳冷凍胚胎植入懷孕率
(vs 不補充 or 重劑量賀爾蒙補充since D2,3)
Several replacement protocols for frozen-thawed ET (FET) exist, with no advantage of one protocol over the others. In the present study, we aim to evaluate the outcome of natural cycle FET with modified luteal support.
All consecutive patients undergoing natural or artificial hormone replacement (AHR) day-2/3 FET cycles between May 2012 and June 2015 in our IVF unit were evaluated. While AHR FET cycles were consistent, those undergoing natural cycle FET received progesterone luteal support, and from June 2014, patients received two additional injections, one of recombinant hCG and the other of GnRH-agonist, on day of transfer and 4 days later, respectively (modified luteal support).
Patients' clinical characteristics and laboratory/embryological variables were comparable between those undergoing natural vs. AHR cycles, during the earlier as compared to the later period. Moreover, while implantation, clinical, and ongoing pregnancy rates were significantly higher during the later period in patients undergoing the natural cycle FET with the modified luteal support (31, 51, and 46 %, respectively), as compared to natural (17, 26, and 20 %, respectively), or AHR FET in the late study period (15, 22, and 17 %, respectively), the natural cycle FET without the additional two injections yielded the same results, as the AHR cycles.
We therefore suggest that in ovulatory patients undergoing FET, natural cycle FET with the modified luteal support should be the preparation protocol of choice. Further large prospective studies are needed to elucidate the aforementioned recommendation prior to its routine implementation.
(vs 不補充 or 重劑量賀爾蒙補充since D2,3)
J Assist Reprod Genet. 2016 Mar 14. [Epub ahead of print]
Natural cycle frozen-thawed embryo transfer-can we improve cycle outcome?
Abstract
PURPOSE:
METHODS:
RESULTS:
CONCLUSIONS:
一般精蟲細胞膜為glycocalyx表面帶負電
約有94%精蟲表面帶負電
可使用微電泳篩顯帶負電之精蟲再進行傳統離心分離精蟲
傳統離心分離精蟲會改變精蟲表面之帶電性
表面帶負電之精蟲進行傳統離心分離後DNA受損率較低
The sperm membrane undergoes extensive surface remodeling as it matures in the epididymis. During this process, the sperm is encapsulated in an extensive glycocalyx layer, which provides the membrane with its characteristic negative electrostatic charge. In this study, we develop a method of microelectrophoresis and standardize the protocol to isolate sperm with high negative membrane charge.
Under an electric field, the percentage of positively charged sperm (PCS), negatively charged sperm (NCS), and neutrally charged sperm was determined for each ejaculate prior to and following density gradient centrifugation (DGC), and evaluated for sperm DNA damage, and histone retention. Subsequently, PCS, NCS, and neutrally charged sperm were selected using an ICSI needle and directly analyzed for DNA damage.
When raw semen was analyzed using microelectrophoresis, 94 % were NCS. In contrast, DGC completely or partially stripped the negative membrane charge from sperm resulting PCS and neutrally charged sperm, while the charged sperm populations are increased with an increase in electrophoretic current. Following DGC, high sperm DNA damage and abnormal histone retention were inversely correlated with percentage NCS and directly correlated with percentage PCS. NCS exhibited significantly lower DNA damage when compared with control (P < 0.05) and PCS (P < 0.05). When the charged sperm population was corrected for neutrally charged sperm, sperm DNA damage was strongly associated with NCS at a lower electrophoretic current.
The results suggest that selection of NCS at lower current may be an important biomarker to select healthy sperm for assisted reproductive treatment.
約有94%精蟲表面帶負電
可使用微電泳篩顯帶負電之精蟲再進行傳統離心分離精蟲
傳統離心分離精蟲會改變精蟲表面之帶電性
表面帶負電之精蟲進行傳統離心分離後DNA受損率較低
J Assist Reprod Genet. 2016 Mar 23. [Epub ahead of print]
Optimization of microelectrophoresis to select highly negatively charged sperm.
Abstract
PURPOSE:
METHODS:
RESULTS:
CONCLUSION:
使用捐卵而懷孕之孕婦懷孕期間併發衽娠毒血症機率為一般IVF孕婦之3倍
The objective of this meta-analysis is to determine whether there is a higher incidence of preeclampsia (PE) in pregnancies achieved by oocyte donation (OD) compared with pregnancies achieved by in vitro fertilization with autologous oocytes (IVF).
A systematic review was performed to identify relevant studies published from January 1994 until April 2015 with at least an abstract in English using PubMed, ISI Web of Knowledge, and clinicaltrials.gov. The 11 studies included in this systematic review were retrospective and prospective cohort studies of women reporting results on the association between oocyte donation vs. in vitro fertilization (exposure) and preeclampsia (outcome).
Oocyte donation is a risk factor for the development of PE compared to IVF cycles, with a weighted OR of 3.12 under a fixed effects method (FEM: no heterogeneity between the studies). The weighted OR under a random effects model was 2.9 (REM: heterogeneity between the studies). The meta-regression analysis showed that neither multiple pregnancies (estimate = 0.08; p = 0.19) nor patient age (estimate = -2.29; p = 0.13) significantly explained the variability of the effect of oocyte donation on PE. Q statistic was 12.78 (p = 0.237), suggesting absence of heterogeneity between the studies.
Pregnancies achieved by oocyte donation confer a threefold increase in the likelihood of developing PE than those achieved by in vitro fertilization with own oocytes. Physicians should be aware of this risk in order to both counsel patients and monitor pregnancies accordingly.
J Assist Reprod Genet. 2016 Mar 23. [Epub ahead of print]
Is oocyte donation a risk factor for preeclampsia? A systematic review and meta-analysis.
Abstract
PURPOSE:
METHODS:
RESULTS:
CONCLUSIONS:
解凍胚胎於2, 5, 20 % O2培養環境囊胚期形成率類似( 58.7% vs 63.6 % vs 66.7 %)
5% O2培養環境下囊胚品質較優
The objective of this study is to investigate the effect of 2, 5, and 20 % O2 on post-thaw day 3 human embryo culture until blastocyst stage.
One hundred fifty-five day 3 human embryos were used. One hundred twenty out of 155 embryos were recovered after thawing. Surviving embryos were distributed into 2, 5, or 20 % O2 groups and cultured for 2.5 days. At the end of culture, blastocyst formation was assessed, and then, embryos were collected for RT-qPCR or immunofluorescence analysis.
Using visible blastocoel to define blastocyst formation, 58.7 % (27/46) of surviving day 3 embryos formed blastocyst at 2 % O2, 63.6 % (28/44) at 5 % O2, and 66.7 % (20/30) at 20 % O2. The difference in blastocyst formation rates was not significant. Average blastocyst cell number was 119.44 ± 11.64 at 2 % O2, 142.55 ± 22.47 at 5 % O2, and 97.29 ± 14.87 at 20 % O2. Average apoptotic rate was 4.7 % ± 0.4 % for blastocyst formed at 2 % O2, 3.5 % ± 0.7 % at 5 % O2, and 5.8 % ± 1.1 % at 20 % O2. Apoptosis rate was significantly lower for blastocysts formed at 5 % O2(p < 0.05). Compared with gene expression levels at 5 % O2, which were arbitrarily set as "1," 20 % O2 is associated with significantly higher expression of BAX (2.14 ± 0.47), G6PD (2.92 ± 1.06), MnSOD (2.87 ± 0.88), and HSP70.1 (8.68 ± 4.19). For all genes tested, no significant differences were found between 2 and 5 % O2.
The result suggests that development of cryopreserved human embryos from day 3 to blastocyst stage benefits from culture at 5 % O2.
5% O2培養環境下囊胚品質較優
J Assist Reprod Genet. 2016 Mar 23. [Epub ahead of print]
Comparison of 2, 5, and 20 % O2 on the development of post-thaw human embryos.
Yang Y1,2,3, Xu Y1,4, Ding C1,4, Khoudja RY1,4, Lin M1,4,5, Awonuga AO2, Dai J2, Puscheck EE2, Rappolee DA2,3, Zhou C6,7.
Abstract
PURPOSE:
METHODS:
RESULTS:
CONCLUSION:
Day5 vs Day6冷凍囊胚植入後活產率無明顯差異 (52.4 vs 52.6 %)
高品質囊胚染色體正常率明顯高於低品質囊胚 (55.3 vs 41.5 %)
高品質囊胚植入後活產率明顯高於低品質囊胚 (54.5 vs 25.0 %)
J Assist Reprod Genet. 2016 Apr 21. [Epub ahead of print]
Whether there are differences in the pregnancy outcomes of blastocysts cryopreserved during different developmental stages remains under debate because the results among studies are inconsistent. We analyzed blastocyst quality and pregnancy outcomes by considering blastocyst euploidy and investigated the differences in the development potential between blastocysts of different developmental stages (frozen-thawed day 5 [D5] and day 6 [D6] cycles) and their relationship with clinical pregnancy outcomes.
In total, 1374 D5 and 255 D6 frozen-thawed blastocyst transfer cycles were retrospectively analyzed. Additionally, the chromosome euploidy and clinical pregnancy rates of 237 blastocysts from 50 pre-implantation genetic diagnosis (PGS) cycles were statistically analyzed. The corresponding euploidy rate and pregnancy outcomes of the D5 and D6 blastocyst transfers were also compared.
The clinical pregnancy rate (47.2 vs 40.0 %; P = 0.04) and implantation rate (34.2 vs 28.8 %; P = 0.03) of the D5 blastocysts were higher than were those of the D6 blastocysts. However, the clinical pregnancy rate (52.4 vs 52.6 %; P = 0.97) and implantation rate (38.9 vs 35.6 %; P = 0.39) of the high-quality D5 blastocysts did not significantly differ from those of the high-quality D6 blastocysts. Analysis of blastocyst euploidy in 237 blastocysts examined in 50 PGS cycles showed that the euploidy rates of the D5 and D6 blastocysts were both 48.1 % (P = 0.99). The clinical pregnancy rate of the D5 blastocysts (48.5 vs 17.6 %; P = 0.03) was higher than that of the D6 blastocysts. The euploidy rates (55.2 vs 55.3 %; P = 0.99) and clinical pregnancy rates (60.0 vs 42.9 %; P = 0.77) of the high-quality D5 and D6 blastocysts did not differ. The euploidy rate (55.3 vs 41.5 %, P = 0.03) and clinical pregnancy rate (54.5 vs 25.0 %, P = 0.03) of the high-quality blastocysts were higher than were those of the poor-quality blastocysts.
The euploidy rates between the D5 and D6 blastocysts did not differ. High-quality D6 blastocysts in frozen-thawed cycles had similar developmental potential and pregnancy outcomes compared to those of high-quality D5 blastocysts. The quality of the blastocysts was an important factor that affected the pregnancy outcomes of the frozen-thawed cycles.
高品質囊胚染色體正常率明顯高於低品質囊胚 (55.3 vs 41.5 %)
高品質囊胚植入後活產率明顯高於低品質囊胚 (54.5 vs 25.0 %)
J Assist Reprod Genet. 2016 Apr 21. [Epub ahead of print]
Comparison of differences in development potentials between frozen-thawed D5 and D6 blastocysts and their relationship with pregnancy outcomes.
Abstract
PURPOSE:
METHODS:
RESULTS:
CONCLUSIONS:
2016年4月29日
卵子之中空透明帶ZP於冷凍解凍過程中具有保護胚胎卵子作用
Mammalian oocytes have a proteinaceous hydrogel-like outer shell known as the zona pellucida (ZP) that semi-encloses their plasma membrane and cytoplasm. In this study, we cryopreserved mouse oocytes either with or without ZP by vitrification. Our results show that the presence of an intact ZP could significantly improve the post-vitrification survival of oocytes to 92.1% from 13.3% for oocytes without ZP. Moreover, there was no significant difference in embryonic development between fresh and cryopreserved oocytes with ZP after in vitro fertilization (IVF). Further atomic force microscopy (AFM) analysis showed that the intact oocytes with ZP have an elastic modulus that is more than 85 times higher than that of oocytes without ZP. This may partially explain the important role of ZP in protecting the oocytes by resisting the mechanical stress due to possible ice formation during cryopreservation by vitrification. Collectively, this study reveals a new biophysical role of ZP during vitrification of oocytes and suggests microencapsulation of the many mammalian cells without a ZP in ZP-like hydrogel is an effective strategy to improve their survival post cryopreservation by vitrification.
Cryobiology. 2015 Oct;71(2):350-5. doi: 10.1016/j.cryobiol.2015.08.012. Epub 2015 Aug 20.
The crucial role of zona pellucida in cryopreservation of oocytes by vitrification.
Abstract
冷凍精蟲抗凍劑比較( egg-yolk citrate +12%抗凍劑)
精蟲存活率 Gly >EG>DMSO
[GLY (77.8 ± 11.0%), EG (20.4 ± 10.1%), DMSO (15.7 ± 11.9%) and PND (11.2 ± 11.3%).]
Cryopreservation has been reported to damage approximately 40-50% of viable sperm in bull semen. The present study was undertaken to assess the cryo-effectiveness of glycerol (GLY), ethylene glycol (EG), dimethyl sulfoxide (DMSO) and propylene glycol (PND) as cryoprotectant during the cryopreservation of Nguni bull semen. Semen was collected from 18 Nguni bulls and evaluated macroscopically and microscopically for sperm parameters. Thereafter, the semen samples were diluted with egg-yolk citrate extender supplemented with either 12% GLY or DMSO or EG or PND cryoprotectant. Semen samples were loaded into straws and placed into a controlled rate programmable freezer and stored in a liquid nitrogen tank. Following semen thawing, artificial insemination (AI) was done on synchronized Nguni cows. The in vitro fertilization (IVF) was conducted on cow's oocytes to test the fertilizing ability. Data was analyzed with the aid of ANOVA. A significant difference (p < 0.05) was recorded between fresh total sperm motility rate (94.7 ± 2.6%) and frozen-thawed sperm total motility rate with GLY (77.8 ± 11.0%), EG (20.4 ± 10.1%), DMSO (15.7 ± 11.9%) and PND (11.2 ± 11.3%). Interestingly, a positive correlation between total sperm motility and pregnancy rate (r = 0.42) was recorded. However, a negative correlation of Nguni sperm parameters with IVF (r = -0.53) was obtained. The freezing-thawing process did reduce the Nguni sperm total sperm motility percentage.
精蟲存活率 Gly >EG>DMSO
[GLY (77.8 ± 11.0%), EG (20.4 ± 10.1%), DMSO (15.7 ± 11.9%) and PND (11.2 ± 11.3%).]
Cryobiology. 2016 Apr 5. pii: S0011-2240(16)30039-6. doi: 10.1016/j.cryobiol.2016.04.001. [Epub ahead of print]
Comparison of four different permitting and combination of two best cryoprotectants on freezing Nguni sperm evaluated with the aid of computer aided sperm analysis.
Abstract
2016年4月27日
VS冷凍液中0.5M sucrose於胚胎冷凍過程中扮演脫水及保護細胞膜之作用
TS解凍液中1M sucrose於胚胎解凍過程中扮演脫出冷凍液EG & DMSO及保護細胞膜之作用
This study was designed to investigate the survival rate of vitrified mouse blastocysts depending on the presence or absence of sucrose in vitrification solution.
Mouse two-cell embryos were collected and cultured to blastocysts. Two vitrification solutions were prepared. The control solution was composed of 25% glycerol, 25% ethylene glycol, and 0.5 M sucrose (G25E250.5S) containing 2.5 mL glycerol, 2.5 mL ethylene glycol, 2 mL SSS, and 0.855 g sucrose in 5 mL PB1. The experimental solution was composed of 25% glycerol and 25% ethylene glycol (G25E25) and contained 2.5 mL glycerol and 2.5 mL ethylene glycol in 5 mL PB1. Artificial shrinkage was conducted by aspirating the blastocoelic fluid using an ICSI pipette. To examine the effect of sucrose in the vitrification solution on the survival rate of mouse blastocysts, the shrunken-equilibrated blastocysts were rehydrated or vitrified after being exposed to one of the two vitrification solutions. After exposure and the vitrification-thawing process, the re-expansion rate and hatching rate were evaluated after 6 hours of in vitro culture.
The re-expansion rate of mouse blastocysts exposed to vitrification solution with and without sucrose were not different in the experimental solution (without sucrose) (98%) and the control solution (with sucrose) (92%) (p>0.05). The hatching rate was higher in the experimental solution (95%) than in the control solution (88%), but did not differ across two treatments (p>0.05). The re-expansion rate of mouse blastocysts vitrified in the control solution was 92% and 94%, respectively (p>0.05), and the hatching rate was higher in the experimental solution (90%) than in the control solution (74%) (p<0.05).
Sucrose need not be added in vitrification solution for freezing of artificially shrunken mouse blastocysts.
TS解凍液中1M sucrose於胚胎解凍過程中扮演脫出冷凍液EG & DMSO及保護細胞膜之作用
Clin Exp Reprod Med. 2014 Sep;41(3):115-9. doi: 10.5653/cerm.2014.41.3.115. Epub 2014 Sep 30.
Vitrification solution without sucrose for cryopreservation in mouse blastocysts.
Abstract
OBJECTIVE:
METHODS:
RESULTS:
CONCLUSION:
大量包覆卵丘細胞之卵子會下降冷凍解凍存活率
保留少量卵丘細胞之卵子(corona radiata)不會下降冷凍解凍存活率
含有卵丘細胞之培養液會提高IVF受孕率
Theriogenology. 2016 Mar 3. pii: S0093-691X(16)00087-X. doi: 10.1016/j.theriogenology.2016.02.015. [Epub ahead of print]
Role of cumulus cells during vitrification and fertilization of mature bovine oocytes: Effects on survival, fertilization, and blastocyst development.
Abstract
2016年4月22日
2-cell胚胎約有42%具多核狀況
2-cell胚胎具多核狀況是可逆性的狀況, 有73%會發展成正常4-cell胚胎
4-cell胚胎約有14%具多核狀況
4-cell胚胎具多核狀況, 胚胎預後較差著床率較低
多核狀況中3-4核之胚胎比2-核之胚胎著床率較低
To study nucleation status in two- and four-cell embryos and its effect on reproductive outcomes.
Retrospective cohort study.
University-affiliated private center.
A total of 1,679 embryos from 940 oocyte donation cycles from May 2012 to May 2014.
None.
Implantation, morphokinetics, and nucleation status restoration.
Multinucleation was present in 42.53% of embryos with known implantation data at the two-cell stage; it was present in approximately 14% of them at the four-cell stage. In all, 73.4% of the embryos were multinucleated at two cells and restored their nucleation status when they cleaved into four cells. Embryos with blastomeres multinucleated (more than two nuclei) at the four-cell stage showed a lower implantation rate. The average length of S-phase in the first embryo cell cycle in the positive known implantation data (KID+) embryos was longer than in KID- (15.50 hours vs. 14.38 hours) and slightly shorter in the second embryo cell cycle (8.35 hours in KID+ vs. 8.60 hours in KID-).
Multinucleation in two-cell-stage embryos is a frequent event, which is reversible in a high proportion of embryos, without impact on the implantation rate; and embryos with multinucleated blastomeres have a reduced outcome compared with those with binucleated blastomeres when multinucleation is present in four-cell-stage embryos.
2-cell胚胎具多核狀況是可逆性的狀況, 有73%會發展成正常4-cell胚胎
4-cell胚胎約有14%具多核狀況
4-cell胚胎具多核狀況, 胚胎預後較差著床率較低
多核狀況中3-4核之胚胎比2-核之胚胎著床率較低
Fertil Steril. 2016 Apr 5. pii: S0015-0282(16)61043-9. doi: 10.1016/j.fertnstert.2016.03.036. [Epub ahead of print]
Study of nucleation status in the second cell cycle of human embryo and its impact on implantation rate.
Abstract
OBJECTIVE:
DESIGN:
SETTING:
PATIENT(S):
INTERVENTION(S):
MAIN OUTCOME MEASURE(S):
RESULT(S):
CONCLUSION(S):
早期多核胚胎仍有75%可能發育成為正常胚胎&順利著床
To assess the developmental competence of human embryos with multinucleation (MN).
Experimental study.
Research institute of private fertility center.
Forty-four couples donating 143 zygotes for confocal imaging study, and 78 couples included in the retrospective clinical study.
Time-lapse imaging study using confocal and light microscopes.
Cytokinesis at first mitosis, MN, chromosomal behavior, euploidy, implantation, successful delivery of healthy baby.
About 25% of the embryos showed abnormal cytokinesis (n = 34). All showed MN, and their development was greatly impaired. More than 75% of embryos that showed normal cytokinesis at first mitosis displayed MN (n = 81). However, the subsequent development of embryos with MN was similar to that of embryos without MN in vitro and in vivo. Most blastocysts were euploid. All chromosomes in several MNs took part in forming a bipolar spindle after the nuclear envelope breakdown followed by normal cleavage and development to the blastocyst stage. The implantation potential of embryos with MN was similar to that of embryos without MN, and healthy babies were born from the former group after transfer.
The presence of MN after the first mitosis does not adversely affect the subsequent development of embryos if they showed normal cytokinesis at this stage. The poor development of embryos with MN is mainly caused by abnormal first cytokinesis.
Fertil Steril. 2016 Apr 6. pii: S0015-0282(16)30058-9. doi: 10.1016/j.fertnstert.2016.03.025. [Epub ahead of print]
Multinucleation per se is not always sufficient as a marker of abnormality to decide against transferring human embryos.
Abstract
OBJECTIVE:
DESIGN:
SETTING:
PATIENT(S):
INTERVENTION(S):
MAIN OUTCOME MEASURE(S):
RESULT(S):
CONCLUSION(S):
訂閱：
文章 (Atom)