Fertilization of IVF/ICSI Using Sibling Oocytes from Couples with Subfertile Male or Unexplained Infertility

The significance of the performance of conventional in vitro fertilization and intracytoplasmic sperm injection (IVF/ICSI) using sibling oocytes from couples with subfertile male or unexplained infertility was evaluated. A total of 410 sibling oocyte cumulus-corona complexes (OCCC) from 21 couples with subfertile male (group A) and 11 unexplained infertile couples (group B) were randomly divided, in order of retrieval, into two groups inseminated either by conventional IVF or by ICSI. The treatment outcomes and the influence of infertility factors on fertilization in each group were compared. The results showed that although the two pronuclear (2PN) fertilization rate per injected sibling oocytes was significantly higher after ICSI (group A: 68.2 %±28.8 %; group B: 66.2 %±24.9 %) than after conventional IVF (group A: 41.8 %±32.7 %; group B: 40.1 %±22.1 %), the other variables studied included: the fertilization rates of per allocated sibling oocytes IVF/ICSI, the fertilization rates of sibling oocytes IVF/ICSI after excluding failed IVF fertilization cycles, as well as the cleavage rates of normal fertilization were not statistically significant (P>0.05). Similarly, though the total fertilization failure rate in the IVF group (group A: 42.9 %; group B: 36.4 %) was significantly higher than in the ICSI group (group A: 4.8 %; group B: 0), we did not cancel cycles due to the normal fertilization of sibling oocytes. Embryo transfer was possible in all 32 couples. There were 10 clinical pregnancies in the two groups. We also discovered a possible association between some semen parameters and sperm functions of group A, and women age and duration of infertility of group B and fertilization. It is suggested that adoption of the split IVF/ICSI technology in the above cases may help eliminate fertilization failures. This is also a useful method to investigate the effect of single factor on the employment of assisted reproductive technology.

Effects of Different Culture Systems on In Vitro Fertilization of Oocyte in Bovine

[ Objective] To investigate the optimal culture system for in vitro fertilization （IVF） of oocyte in bovine. [ Method] The IVF of mature bovine oocytes was conducted and the fertilized eggs were cultured in vitro with different culture systems. In the traditional culture system, the TALP medium and BO medium were used as fertilization medium, respectively; and M-199 medium supplemented with 5% FCS was used as fertilized egg culture medium. In the transitional culture system, the TALP medium and BO medium were used as fertilization medium; 60% TALP and 40% M-199 supplemented with 5% FCS and 60% BO and 40% M-199 supplemented with 5% FCS were used as fertilized egg culture medium, respectively. The effects of different culture systems on fertilization rate and cleavage rate were observed. E Result] In the transitional culture system （with TALP medium as fertilization medium and with 60% TALP and 40% M-199 supplemented with 5% FCS as fertilized egg culture medium）, the fertilization rate and cleavage rate were increased to 77.8% and 55.6%, respectively. [ Conclusion] The transitional culture system （with TALP medium as fertilization medium and with 60% TALP and 40% M-199 supplemented with 5% FCS as fertilized egg culture medium） is the optimal culture system for IVF of bovine oocyte.

Evaluation of pig oocyte in vitro maturation and fertilization using three gonadotropin-based hormonal compounds

Objective:To evaluate the effect of human chorionic gonadotropins(hCG)and equine chorionic gonadotropins(eCG)on in vitro gilt oocyte maturation and embryonic development,using frozen semen for fertilization.Methods:Two independent experiments(6 replicates each)were carried out to evaluate gilt oocyte maturation,and fertilization and embryonic development by using ovaries from a local abattoir.Totally,712 oocytes were randomly distributed in four-well dishes to receive Novormon(eCG 5.0 IU),PG600(eCG 5.0 IU and hCG 2.5 IU),Chorulon(hCG 5.0 IU),or no hormones.Oocytes were incubated with 5%CO2,95%air and saturation humidity at 39℃for 44 h.Maturation of the oocytes to metaphaseⅡwas assessed by using the aceto-orcein technique.In addition,741 oocytes were used and randomly distributed in four-well dishes,and then oocyte maturation was carried out as mentioned,but matured oocytes were washed and placed in fertilization medium with frozen-thawed sperm.Gametes were co-incubated for 7 h,and then washed and placed in development medium,and incubated for further 7 days,at which time embryonic development was evaluated.Fertilization and embryo development media were not supplemented with the studied hormones.Results:Novormon(eCG)and PG600(eCG+hCG)treatments significantly improved the percentages of metaphaseⅡoocytes compared to the control group(P<0.05).Furthermore,a significant increase was also observed in the young blastocyst stage between the control group and the PG600 treatment group(P<0.05).Conclusions:Hormonal products Novormon(eCG)and PG600(eCG+hCG)can obtain the highest percentages of in vitro maturation in gilt oocytes;however,this effect is not transferred to fertilization rates.

Relationship between lower number of oocytes retrieved and clinical outcomes of in vitro fertilization-embryo transfer

Objective:To explore the clinical outcomes of the infertile women with retrieved oocytes less than or equal to 5 undergoing in vitro fertilization-embryo transfer（IVF-ET） or intracytoplasmic sperm injection（1CSI）. Methods:The clinical data of 216 embryo transfer cycles with retrieved oocytes less than or equal to 5 during the procedure of IVF/ICSI in Reproductive Medicine Center of the 105th Hospital of PLA from Jul.2008 to Dec.2011 were analyze retrospectively.All the patients were divided into group A（< 35 years）,group B（35-39 years） and group C（≥40 years） according to the ages,and 409 IVF/ICSI cycles with patients’ age less than 35 years old and 6-15 retrieved oocytes in the same period were served as controlled group.Then the patients≥35 years were subdivided into gonadotropin-releasing hormone agonist（GnRH-a） long protocol group,GnRH-a short group and GnRH antagonist group according to the protocols of controlled ovarian hyperstimulation（COH）.The clinical date and the outcomes were analyzed and compared among all groups. Results:There were significantly differences in clinical pregnancy rate（38.3%vs.19.4%） and early abortion rate（16.1%vs.50.0%） between group A and group C（P<0.05）,and there were no significant differences in clinical pregnancy rate（38.3%vs.41.6%）and early abortion rate（16.1%vs.10.0%） between group A and control group（P>0.05）.There were no significant differences in clinical pregnancy rates（29.01%vs.26.1%vs.25.9%） and early abortion rates（33.3%vs.33.3%vs.40.0%） among GnRH-a long protocol group,GnRH-a short group and GnRH antagonist group（P>0.05）. Conclusions:Relatively satisfactory clinical outcomes of IVF/ICSI would still be got for the patients <35 years with retrieved oocytes less than or equal to 5,but whatever COH protocols such as GnRH-a long protocol,GnRH-a short and GnRH antagonist could not improve the outcomes of IVF/ICSI for the patients aged≥35 with retrieved oocytes less than or equal to 5.

Influence of Follicular Fluid on <i>in Vitro</i>Maturation and Fertilization of Bovine Oocytes

The aim of this study was to investigate the effect of time on in vitro maturation of bovine oocytes and of the addition of follicular fluid on meiotic progression. The cumulus-oocyte complexes (COCs) collected from 3 to 6 mm follicles were obtained from ovaries of slaughtered female animals. The medium of maturation was supplemented or not with 20 μL follicular fluid (FF);661 oocytes were matured in vitro (extrusion of the first polar corpuscle) for 22 hours with added follicular fluid (AFF) (72.01%) or without follicular fluid (WFF) (67.53%) and 679 oocytes were matured in vitro for 26 hours (extrusion of the first polar corpuscle) with AFF (92.1%) and WFF (77.15%). The results of extrusion of the second polar corpuscle as an event related to the fertilization percentages showed that the increase in the fertilization rate is maintained at 26 hours with AFF (79.45%), but the percentage decreases WFF (65.08%). After 22 hours, the fertilization rate was 62.38% AFF and 53.40% WFF. The developmental competence of bovine oocytes is affected by the duration of maturation in vitro and the inclusion in the FF culture medium. The use of follicular fluid in the in vitro maturation medium may be a biological strategy to increase the cumulus expansion, the nuclear maturation and the in vitro fertilization.

Effects of Epidermal Growth Factor and β-mercaptoethanol on In-vitro Fertilization and Development of Oocytes from Hormone-Stimulated Lambs

[Objective]The aim was to explore optimization of system of oocytes in-vitro culture of young animals. [Method]Effects of EGF （ epidermal growth factor） and β-mercaptoethanol in maturation media on fertilization,cleavage and blastaea were researched,which were then compared with those of adult sheep. [Result]EGF in different concentrations had little effects on rates of cleavage and blastaea （ P 〉0.05） and β-mer-captoethanol in different concentrations would improve blastaea rate of oocytes,for example,100 μmol/L of β-mercaptoethanol has significant effects on balstaea rate （ P 〈0.05） ,but has little effects on cleavage rate （ P 〉0. 05） . In addition,rates of cleavage and balstaea of oocytes in lamb were both lower than those of adult sheep （ P 〈0.05） ; fertilization rate of oocytes in lamb （ P 〈0.05） ,which differed little with that of adult sheep （ P 〉0.05） ,could be significantly enhanced by 100 μmol/L of β-mercaptoethanol. Furthermore,polyspermy rate was higher than that of adult sheep without β-mercaptoethanol （ P 〈0.05） ; the rate was of little differences with that of adult sheep with 100 μmol/L of β-mercaptoethanol （ P 〉 0.05） ; unfertilization rate （ 20%） in media without β-mercaptoethanol was a little higher （ P 〉0.05） than those of adult sheep （ 12.3%） and those in media with β-mercaptoethanol （ 13.5%） . [Conclusion]Developmental capacity of oocytes and fertilization rate could be improved by 100 μmol/L of β-mercaptoethanol with polyspermy rate reduced,but developmental capacity of lamb was significantly lower than that of adult sheep.

Comparative Study on Activation and Fertilization of Mouse Oocytes at Different Ages

Comparisons of activation rates and fertilization rates were made among oocytes at different ages. Results showed that oocytes at different ages had different activation and fertilization rates when stimulated by sperm or ethanol. Oocytes at 15～24 h after the injection of hCG were readily activated by 8% ethanol. The activation rate of oocytes increased with the age of oocytes, up to the highest average of 81.6%, but decreased after 20 h posthCG. Oocytes at 20 h posthCG exhibited the highest immediate cleavage rate(48.0%) after being stimulated by ethanol. On the other hand, 13～15 h oocytes exhibited higher fertilization rates, and the older oocytes were more difficult to be fertilized by sperm in vitro. These suggest that oocytes can be activated in different ways; the mechanism of fertilization might be different from that of activation; and in vitro fertilization is more dependent on oocyte age.

LH Plays a Role to Enhance the Nuclear and Cytoplasm Synchronization During In vitro Maturation of Ovine Oocytes

This study is aimed to investigate the effect of luteinizing hormone （LH） on maturation and fertilization in vitro of ovine oocytes. The ovine oocytes were cultured in vitro in medium with or without LH, and then checked by confocal laser scanning microscope to observe the distribution of cortical granules stained with FITC-LCA during different maturation periods. Similarly, some in vitro matured oocytes were also fertilized in vitro for analysis of their developmental potentiality further. After being cultured in vitro for 4 h, there were significant differences about the rate of germinal vesicle break down （GVBD） between the treatment （with LH） and the control groups （without any hormones） （36.76% vs 18%, P〈0.05）. Further, there were also significant differences of the cleavage and blastocyst rates between these two groups （67.15% vs 42.37%, 21.9% vs 12.71%, P〈0.05, respectively）. The distribution of cortical granules appeared to spread from the edges to the central site of sheep oocytes following their delaying durations of maturation in vitro. It can be concluded that LH may play a role to delay the occurence of GVBD, prolong the maturation duration of cytoplasm, and enhance the nuclear and cytoplasm synchronization of ovine oocytes matured in vitro and finally improve their in vitro developmental potentiality.

In vitro maturation of human oocytes maintaining good development potential for rescue intracytoplasmic sperm injection with fresh sperm

BACKGROUND The outcomes of the use of commercial in vitro maturation(IVM)medium to culture immature oocytes obtained from conventional ovulation induction,followed by rescue intracytoplasmic sperm injection(RICSI),are not ideal.It is thus difficult to widely adopt this approach in clinical practice.Therefore,it is necessary to explore methods for improving the clinical outcome of IVM.AIM To study the effect of sperm on the developmental potential of in vitro-matured oocytes in conventional culture.METHODS This was a retrospective study of patients whose immature oocytes were harvested from conventional oocyte stimulation cycles and underwent ICSI at our hospital between June 2018 and August 2020.RICSI was performed using sperm collected on the day of oocyte harvest(old)and sperm collected on the day of RICSI(fresh)and oocytes matured in vitro after 24 h of culture in conventional medium.The rates of in vitro oocyte maturation,normal fertilization,normal cleavage,day-3 top-quality embryos,and useful blastocyst formation were compared between the two groups.RESULTS In total,102 germinal vesicle(GV)-stage immature oocytes were cultured in the old sperm group.In the fresh sperm group,122 GV-stage immature oocytes were collected and cultured in vitro for 24 h.There were no significant differences in the general conditions of males and females between the two groups(P>0.05).The oocyte maturation,normal fertilization,and normal cleavage rates of the old and fresh groups were 51.0%vs 55.7%,61.5%vs 64.7%,and 93.8%vs 93.2%,respectively.None of the rates differed significantly(P>0.05)between the two groups.However,the day-3 top-quality embryo and useful blastocyst rates of the old and fresh sperm groups were 16.6%vs 63.4%;6.67%vs 34.6%,respectively.The day-3 top-quality embryos and useful blastocyst rates of the old sperm group were significantly lower than those of the fresh group(P<0.05).CONCLUSION In vitro maturation with conventional culture medium combined with the use of fresh sperm collected on the day of RICSI is an easy-to-implement strategy for patients whose oocytes are completely or mostly immature.

L-carnitine improves developmental competence of buffalo oocytes in vitro

Objective:To investigate the effect of L-carnitine on in vitro maturation and subsequent in vitro embryo production of buffalo oocytes.Methods:Cumulus oocyte complexes(COCs)were aspirated from ovaries of slaughtered buffaloes.COCs were classified into good and fair qualities based on morphological observation of numbers and integrity of cumulus cells surrounding the oocyte.Both categories of COCs were placed in in vitro maturation medium with supplementation of different concentrations(0,0.250,0.375 or 0.500 mg/mL)of L-carnitine.Oocytes from both qualities were in vitro fertilized and in vitro cultured for 7 days,to examine the developmental competence.Results:Supplementation of L-carnitine to in vitro maturation medium increased the cumulus cell expansion rate of COCs to grade A,and reduced the cumulus cell expansion of COCs to grade B and grade C in both good and fair quality oocytes.Similarly,L-carnitine induced the in vitro meiotic progression of buffalo oocytes to metaphaseⅡin both good and fair quality oocytes.Additionally,L-carnitine reduced the rate of oocyte degeneration in both good and fair quality oocytes.L-carnitine increased the rate of cleaved formation at day 2 and blastocyst formation at day 7 during in vitro culture in both qualities of oocytes.Moreover,a higher rate of blastocyst production was observed in L-carnitine-treated fair quality oocytes,which was higher than the results in the untreated good quality oocytes.Conclusions:L-carnitine enhances meiotic maturation and subsequent embryo development from both good and fair quality buffalo oocytes.

A modified protocol for <i>in vitro</i>maturation of mouse oocytes from secondary preantral follicles

A 2-step culture system was designed and tested for the in vitro maturation efficiency of oocytes from pre-puberty preantral follicles of FVB/N inbred mice. The following modifications were made: 1) The concentration of ITS was reduced by half in the basal MIF medium to minimize uncoordinated growth between oocyte and GC cells;2) Heterogeneous preantral follicles were cultured in groups of 3 - 5 follicles in hanging drops of medium with reduced concentration of ITS for six days to induction follicular aggregation. This hanging drop method mimics a 3-D IVM culture system at the early stage of cultivation in which the sphere structure of each follicle is well maintained. It also enables follicles in each aggregate to communicate with each other, synchronize their growth, and thus prevent immature follicular rupture. 3) Medium was further supplemented with retinoic acid to enhance developmental capacity of meiotically arrested oocytes. After a 14-day culture in vitro, ~37% of the collected inbred preantral follicles completed nuclear maturation. Approximately 94% of the mature oocytes tested were able to be fertilized;and 77% of them developed into healthy embryos. These results demonstrate that our IVM system is reliable to produce a satisfactory number of high quality oocytes. In addition, multiple cytoplasmic parameters, including gene expression of key regulators, chromosome/spindle organization, mitochondrial proliferation and distribution, and total ATP content were explored to characterize the supportive and limiting components of our IVM system so that the culture system can be further optimized.

Endometrial Ice Application Prior to Embryo Transfer in Cases of Recurrent Unexplained <i>In-Vitro</i>Fertilization Failure: A Pilot Study

Introduction: This prospective pilot study was done to get preliminary data about a new technique of inflecting endometrial injury in order to improve implantation rate in IVF cases. Patients and methods: 60 patients were en-rolled and randomly distributed into 3 groups. Each group included 20 pa-tients. This study was conducted in the Egyptian consultants for infertility management and IVF center Tanta-Egypt, in the period from January 2015 to March 2016. In group A (study group 1), the 20 patients included in this group underwent endometrial scratching once at day 21 of menstrual cycle in the cycle prior to ET by a 4 mm disposable Karman’s cannula inserted through the cervical os. While in group B (study group 2), the 20 patients underwent the new technique of endometrial Ice bars application for 10 minutes at day 7 of the embryo transfer cycle to inflect the physical trauma required for the change in the endometrial receptivity. In Group C (control group), the 20 patients didn’t receive any extra action rather than the ordinary preparation for embryo transfer. The primary outcome measures of the study were the clinical pregnancy rate and the number of pregnancies that exceeded 14 weeks of gestation. The secondary outcome measures were multiple pregnancy and miscarriage ratios. Results: The results of this pilot study shows a (>14 weeks) pregnancy rate which is equal in the endometrial icing (30% n = 6/20) and endometrial scratching group (30% n = 6\20). The >14 weeks pregnancy rate in both groups is higher than the control group (25% n = 5/20). The small number of patients in the pilot study may explain the difference to be statistically non-significant. The abortion rate was 33.3% (n = 3/9) in the scratching group and 25% (n = 2/8) in the icing group and 16.6% (n = 1/5) in the control group. The multiple pregnancy rate was 16.6% (n = 1/6) in the scratching group and 33.3% (n = 2/6) in the icing group and 20% (n = 1/5) in the control group. Conclusion: Endometrial icing is a new technique of endometrial injury which deserves more studies to test its efficacy in improving embryo implantation and to test its safety.

The influence of macrophage co-culture to the fertility rate and blastocyst formation rate of the mouse's oocytes

Objective:To investigate the effect of co-culture with peritoneal macrophage to the fertility rate and blastocyst formation rate of the mouse's oocytes.Methods:A total of 15 KunMing mice aged 10-14 weeks were undergone ovulation induction and oocytes collection.The oocytes were randomly divided into three groups.The first group was the control group and cultured without macrophage;the second group(experimental group 1)was co-cultured with macrophages right after oocytes collection;the third group(experimental group 2)was cultured alone after oocytes retrieval,and co-cultured with macrophages from 8 cell stage.Results:1.The fertility rate in control group(91.0%)and experimental group 2(93.7%)was significantly higher than that in experimental group 1(72.0%,P<0.001).2.Blastocyst formation rate of the fertilized oocytes in the experimental group 2(78.2%)was much higher than control group(60.3%,P<0.001).3.Blastocyst formation rate of the oocytes in experimental group 2(73.2%)was much higher than control group(54.8%)and experimental group 1(50.0%),which were statistically significant.Conclusions:1.Co-culture with peritoneal macrophages had an adverse influence on the fertilization process.2.Peritoneal macrophages could enhance the mouse embryonic development and increase the blastocyst formation rate.

Case Report <i>ex Vivo</i>Oocyte Collection to Maintain Fertility in a Patient with Micropapillary Serous Borderline Ovarian Tumor

Background: Approximately 11% of cancer cases are diagnosed in people of childbearing age. Borderline ovarian tumors (BOTs) make up 10%-15% of all ovarian epithelial malignancies. More than one-third of all BOTs occur in women under 40 years of age. Maintaining the fertility of cured patients is the common goal of both oncologists and reproductologists. Aim: Giving young women diagnosed with a prognostically worse type of BOT and after bilateral adnexectomy the possibility to have their genetically own children by the method of ex vivo oocyte collection. Case Presentation: A 34-year-old nulligravid woman with BOT underwent right laparoscopic salpingo-oophorectomy. Histologically, a serious borderline tumor with a micropapillary pattern and a tumor locus on the ovarian surface were found. Due to histopathology, the oncologist recommended re-staging surgery: laparotomy, left salpingo-oophorectomy, omentectomy and hysterectomy. The patient refused a hysterectomy as she was planning to get pregnant with her partner. To maintain her fertility, controlled hormonal hyperstimulation and ex vivo aspiration of follicles from the ovary after salpingo-oophorectomy was performed. Ex vivo follicle expiration yielded 10 oocytes. 9 mature oocytes were fertilized by ICSI. The 6 embryos of the highest quality were individually frozen by vitrification. Cryoembryotransfer will be scheduled with the consent of the oncologist. Conclusion: This method is suitable for young women with BOT after bilateral salpingo-oophorectomy in whom ex vivo oocyte collection prevents possible leakage of tumor cells into the abdominal cavity, unlike during the conventional in vivo collection prior to surgery.

Role of ICSI in Non-male Factor Cycles as the Number of Oocytes Retrieved Decreases from Four to One

This study aimed to investigate whether intracytoplasmic sperm injection（ICSI） shows an advantage over in vitro fertilization（IVF） in non-male factor cycles as the number of oocytes retrieved decreases from four to one.We undertook a retrospective analysis of 1305 IVF/ICSI cycles of non-male factor in which four or fewer oocytes were retrieved.Comparisons were made between conventional IVF（CI） and ICSI when one,two,three or four oocyte（s） were retrieved.Primary outcomes including normal fertilization rate,proportion of embryos per obtained oocyte,cycle cancellation rate,implantation rate,clinical pregnancy rate（PR）,live birth rate（LBR）,cumulative PR and cumulative LBR were evaluated.The results showed that the normal fertilization rate（72.5% vs.50.0%） and the proportion of embryos per obtained oocyte（72.5% vs.55.0%） were significantly increased in one oocyte retrieved cycles in ICSI group as compared with CI group.However,the proportion of embryos per obtained oocyte was markedly decreased in ICSI group when three（52.3% vs.61.3%） or four（56.9% vs.64.0%） oocytes were retrieved.The implantation rates,clinical PRs,LBRs,cumulative PRs and cumulative LBRs in CI group were comparable to those in ICSI group when one,two,three or four oocyte（s） were retrieved.In conclusion,ICSI doesn＇t show advantages over IVF in low oocyte yield cycles of non-male factors,even when only one oocyte was retrieved.Key words

Studies on the in vitro fertilizaion in cattle

The in vitro maturation, fertilization and development of bovine ovary oocytes in two differentcultural systems A and B were studied under the conditions of 38.5℃,5%CO2,95%air and 100%humidity.The maturation rates were 94.5%and 91.3%,respectively,and the difference wasextremely significant.Frozen semen were thawed and sperm were capacitated with three kinds ofcapacitation agents for fertilization.The pronucleus rates were 76%,65%-68%and 62%respectively.The rates of embryos developed to morula and blastocyst were 19%,16% and 17%respectively.The developmental rates of embryos cocultured with bovine oviductal epithelium cellsand bovine granulosa cells were 25% and 23.4% respectively,with no significant difference. Freshembryos were transplanted into 15 recipicns,and three of them were pregnant and calves were bornin 1990 and 1991.The pregnant rate was 20%.The emryos developed faster before 8-cell stage andslower after 8-cell stage,in vitro than in vivo.

拮抗剂方案中HCG扳机日直径16~18 mm卵泡比例对PCOS患者IVF-ET结局的影响

目的探讨拮抗剂方案中人绒毛膜促性腺激素(HCG)扳机日直径16~18 mm卵泡比例对接受拮抗剂方案的多囊卵巢综合征(PCOS)患者体外受精-胚胎移植(IVF-ET)结局的影响。方法回顾性分析2016年1月至2021年12月于南京医科大学附属妇产医院行IVF-ET助孕的PCOS患者的临床资料。根据HCG扳机日双侧卵巢直径16~18 mm卵泡比例以三分位法将患者分为三组,该比例<0.43为A组(262例),0.43~0.56为B组(267例),≥0.56为C组(247例)。比较三组一般临床资料、超促排卵周期情况、胚胎发育情况、体外受精周期后首次冻融胚胎移植(FET)情况以及妊娠、围产期结局。结果B组基础促黄体生成素(LH)、基础LH与基础促卵泡激素(FSH)比值显著高于A组和C组,差异有统计学意义(P<0.05)。A组促性腺激素(Gn)使用总量高于B组和C组,HCG扳机日雌二醇(E_(2))水平低于B组、C组,差异有统计学意义(P<0.05)。B组和C组获卵数、2原核(PN)数、卵裂数、囊胚形成数以及可移植胚胎数高于A组,差异有统计学意义(P<0.05)。C组囊胚形成率高于A组和B组,高评分囊胚率高于B组,差异有统计学意义(P<0.05)。三组移植胚胎数、移植胚胎天数、内膜准备方案及内膜厚度比较差异无统计学意义(P>0.05)。三组胚胎着床率、生化妊娠率、临床妊娠率、流产率、活产率、妊娠期糖尿病发生率及妊娠期高血压发生率等比较差异无统计学意义(P>0.05)。结论在拮抗剂方案中,HCG扳机日直径16~18 mm卵泡比例可反映卵母细胞发育潜能,当该比例≥0.56时可获得较好的获卵数、2PN数、卵裂数、囊胚形成数以及可移植胚胎数,在一定程度上可作为PCOS患者HCG扳机时机的参考指标,但该比例并不影响后续首次FET的妊娠结局。

Dynamic Changes of β Tubulin during the Resumption of Meiosis of Mouse Oocyte

Morphological changes of tubulin during the resumption of meiosis in both mouse oocyte and fertilized egg were revealed by indirect immunofluorescent marking with monoclonal antibody against β tubulin. During germinal vesicle period (GV), tubulin was found to be distributed around the GV menibrane. With the disruption of GV membrane, microtubule complexes (MTCs) appeared in cytoplasm, first around GV membrane later to spread to other portions as well. Quantitative difference was noted among different oocytes. MTCs coexisted with spindlesformed by prometaphase tubulin, while metaphase tubulin polymerized into spindles and anaphase and telophase tubulin was concentrated in the two poles of the meiotic apparatus and the midbody. In egg arrested in the 2nd metaphase, whether maturing in vitro or vivo, all the tubulin went to form spindles with no MTCs left in the cytoplasm. After fertilization in vitro,MTCs reappeared in the egg cytoplasm activated by sperms while no MTCs could be revealed in cytoplasm after formation of pronucleus. As demonstrated by this experiment,cytoplasmic tubulin in eggs are polymerized chiefly into two forms: the star-shaped MTCs and the spindle.Cytoplasmic MTCs are the structure newly formed when the 1st and 2nd oocytes resumed meiosis. With colchicine disrupting the polymerization of tubulin, the maternal chromosomes,instead of orderly arrangement and orderly separation, either formed disordered mass or were divided into multiple chromatin masses. However, the penetration of sperm into egg, and decondensation and formation of pronuclei were not affected.

TNF-α、IL-17与PCOS不孕患者IVF-ET助孕结局的关系及交互作用分析

目的探讨肿瘤坏死因子-α(TNF-α)、白细胞介素-17(IL-17)与多囊卵巢综合征(PCOS)不孕患者体外受精-胚胎移植(IVF-ET)助孕结局的关系及其交互作用。方法回顾性分析2022年1月至2023年12月北京市大兴区人民医院收治的126例PCOS不孕患者的临床资料,患者均给予IVF-ET治疗,将IVF-ET 35 d后患者宫内妊娠且有胎心搏动者纳入临床妊娠组,否则纳入未妊娠组。分析影响PCOS不孕患者IVF-ET助孕结局的因素,分析TNF-α与IL-17的交互作用对PCOS不孕患者IVF-ET助孕结局的影响,分析TNF-α、IL-17对PCOS不孕患者IVF-ET助孕结局的预测价值。结果临床妊娠患者62例,未妊娠患者64例。Logistic回归分析显示优质胚胎率(OR=0.331,95%CI:0.148~0.738)、TNF-α水平(OR=3.494,95%CI:1.438~8.491)、IL-17水平(OR=3.144,95%CI:1.281~7.569)是影响PCOS不孕患者IVF-ET助孕结局的因素(P<0.05)。TNF-α与IL-17对PCOS不孕患者IVF-ET助孕结局存在正向交互作用,两者交互助孕结局效应为两者单独作用之和的1.384倍(95%CI:1.012~1.894)。ROC曲线分析显示,TNF-α、IL-17及两者联合预测PCOS不孕患者IVF-ET助孕结局的AUC分别为0.713(95%CI:0.610~0.816)、0.707(95%CI:0.594~0.821)、0.883(95%CI:0.815~0.952)。结论TNF-α、IL-17与PCOS不孕患者IVF-ET助孕结局有关,二者在不良助孕结局中具有正向交互作用,其联合预测助孕结局效能良好。

Oocyte Maturation and Fertilization in vitro in Rhesus Monkey (Macaca mulatta)

It is not clear how oocyte maturation, embryo development and differentiation are regulated, though they are critical to the mechanism of reproduction, in vitro fertilization has been greatly successful in domestic animals and human since the ability of immature mammalian oocytes of undergoing maturation in vitro was first demonstrated by Pincus and Enzmann. Although human in vitro fertilization has experienced immense popularity and more than 5000 test-tube babies have been born to date, serious legal and