The mouse eggs in the various stages, of the development prior to implantation were collected and measurements were made on both the largest and smallest diameters of the vitellus, inner and outer surface of the zona pellucida. The various stages of development used were ovarian oocytes (germinal vesiA®e stage), ovulated but unfertilized egg, ovulated and fertilized egg, the 2-cell embryo on the second day of pregnancy, 4-8-cell embryo on the third day of pregnancy and morulablastocyst on the fourth day of pregnancy, A further comparative study on unfertilized and fertilized tubal eggs was made, The time of l2 hours after H.C.G. injection was chosen as the starting point from which to follow the collection of eggs every 3 hours for 24 hours. Since the volume gives a better comparison of size than diameter, the volume of the total eggs, intrazonal cavity, perivitelline space and the various were calculated for the various preimplantation stages of mouse egg. The volume of zona pellucida was also calsulated by subtraction of the volume of the inner zonal cavity from the volume of total egg and compared with the zona pellucida thickness. All calculations were made by computor(CEIR Time-sharing Computor). The diameter and volume of the vitellus in the ovarian oocyte is the largest one of any stage during the preimplantation stages of development, while the total volume of the entire egg as determined from the diameter of outer surface of the zona pellucida of the ovarian cocyte is the smallest one of any stage during development. The diameter and total volume of the entire egg increases from the ovarian oocytes to the first day of pregnancy and then gradually decreases until the third day of pregnancy. An increase in these parameters again takes place on the fourth day of pregnancy. The zona pellucida of the tubal ova is thicker than that of the oocyte, with the zona pellucida of the fertilized egg being definitely thinner when compared with unfertilized eggs. This phenomenon of decreased thickness in fertilized egg may be associated with zona reaction. The perivitelline space between the vitellus and zona pellucida thus formed following ovulation occupied approximately 40 percent of the total volume enclosed by the inner surface of the zona pellucida (intrazonal cavity) in the 1-cell tubal ova. Neither the cause of the rapid accumulation of fluid after ovulation which resulted in the production of the perivitelline space nor the actual time of the formation of the perivitelline space are known. Some possible reasons for the formation or origin of the perivitelline space are discussed. The size and shape of the vitellus undergo compartive reduction during preimplantation stages of development. The possible reason for the reduction of vitelline volume are discussed.
Animal ; Blastocyst ; Embryo/cytology* ; Embryo Implantation ; Embryo and Fetal Development ; Female ; Fertilization ; Mice ; Ovulation ; Ovum/cytology ; Ovum/growth & development ; Temperature ; Time Factors

The ultrastructural changes of mitochondria in the ovarian oocytes from Graafian follicles, the ovulated tubal ova, and the various stages of preimplantation rabbit embryos have been observed with an electron microscope. From the ovarian oocytes to the 4-cell stage, mitochondria showed oval and round forms with a few cristae arranged concentrically and peripherally at the inner membrane. In 8-cell and 16-cell stages, mitochondria tended to change their forms to be elongated, and their sizes, and the outer membrane of the mitochondria had a tendency to become rough and irregular although there were few changes in the inner structure. In morula, some mitochondria began to show several transverse cristae proceeding into the matrix. Mitochondria rapidly increased in number at the late blastocyst stage. Matrix of mitochondria with transverse cristae found in the morula and in blastocyst stages was less dense than that of the earlier stages. The authors believe that the morphological changes of mitochondria during early embryonal development indicate the level of enzymatic activity at which this organelle is engaged in energy metabolism.
Animal ; Cell Membrane/ultrastructure ; Embryo/ultrastructure* ; Embryo Implantation ; Female ; Microscopy, Electron ; Mitochondria/ultrastructure* ; Organoids/ultrastructure ; Ovum/ultrastructure ; Rabbits

OBJECTIVETo explore the developmental potential of embryos at different developmental days and provide evidence for blastocyst culture of non-top quality cleavage stage embryos in frozen-thawed embryo transfer (FET) cycles.

METHODSThe clinical data of 687 FET cycles were retrospectively analyzed. According to the embryo freezing time, the patients were divided into day 5 (D5) blastocyst group (n=87), day 6 (D6) blastocyst group (n=111) and day 3 cleavage-stage embryo (D3) group (n=489) with hormone replacement cycles or natural cycles for endometrial preparation. The clinical pregnancy rates, miscarriage rates, and implantation rates were compared between the 3 groups.

RESULTSThe clinical pregnancy rate, miscarriage rate and implantation rate per transfer were 58.6%, 9.8%, and 42.9% in D5 group, 32.4%, 19.4%, and 23.3% in D6 group, and 44.9%, 16.4%, and 26.9% in D3 group, respectively. The clinical pregnancy rate and implantation rate were significantly higher in D5 group than in the other two groups (P<0.05).

CONCLUSIONThe D5 blastocysts derived from non-top quality D3 embryos after cryopreservation can have better clinical outcomes than those derived from D3 cleavage-stage embryos and D6 blastocysts, and are therefore a better option than D3 cleavage-stage embryos in FET cycles.

Abortion, Spontaneous ; Blastocyst ; Cleavage Stage, Ovum ; Cryopreservation ; Embryo Implantation ; Embryo Transfer ; Female ; Humans ; Pregnancy ; Pregnancy Rate ; Retrospective Studies

OBJECTIVETo investigate the effect and explore the mechanism of Bushen Antai Recipe (BAR) on pregnancy rate and number of implantation site in blastocyst implantation dysfunction (BID) mice induced by indomethacin.

METHODSPregnant mice were divided into 3 groups randomly: the normal group, the model group and the BAR group. Tap water was given orally to the rats in the normal and model groups, and BAR to the rats in the BAR group from the first day of pregnancy for 5 or 8 days; on the 3rd and 4th day dissolvent was injected subcutaneously twice per day in the normal group, while indomethacin (4.33 mg/kg) was injected subcutaneously twice per day in the other two groups to establish implantation dysfunction model; serum estrogen (E) and progesterone (P4) levels were detected on the 5th and 8th day; the pregnancy rate and number of implanted site was observed and the receptors of E and P4 in endometrium of uterus were examined by immunohistochemistry on the 8th day.

RESULTSThe pregnancy rate and number of implanted site was 27.3% and 5.3 +/- 0.7 respectively in the model group, significantly lower than those in the normal group (90.9%, 13.3 +/- 2.8), and the BAR group (72.7%, 10.7 +/- 2.2, P < 0.05). Serum E level was higher in the BAR group than that in the model group on the 5th and 8th day, and even higher than that in the normal group on the 8th day; serum P4 level was lower in the model and BAR groups than that in the normal group on the 5th day (P < 0.01), but higher in the BAR group than that in the model group on the 8th day. Immunohistochemical observation showed that expressions of E and P4 receptor increased remarkably in the BAR group than those in the model group.

CONCLUSIONBAR increases the pregnancy rate and number of implanted site of indomethacrne induced BID mice through regulating E and P4 levels and enhancing the expressions of their receptors in the endometrium.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Embryo Implantation ; drug effects ; Embryo Implantation, Delayed ; drug effects ; Estrogens ; blood ; Female ; Indomethacin ; Mice ; Pregnancy ; Progesterone ; blood ; Random Allocation

OBJECTIVETo assess the effects of the nuclear status of day 2 preembryos on day 3 embryo quality and implantation potential and to weigh its clinical value in the human in-vitro fertilization-embryo transfer (IVF-ET) program.

METHODSEmbryos obtained from 409 fresh conventional IVF-ET/ICSI cycles from July to October 2006 were assessed retrospectively. Day 2 preembryos were classified according to the number of nuclei in each blastomere in 3 groups: grade A with only mononucleated blastomeres, grade B with one or more blastomeres containing no visible nucleus, and grade C with one or more multinucleated blastomeres. Comparisons were made of the rates of arrested embryos and excellent embryos on day 3 as well as of the pregnancy outcome and implantation potential of those in whom cohorts of similar nuclear scoring embryos were transferred.

RESULTSThere were fewer arrested embryos and more excellent embryos on day 3 in grade A than in grade B and C (P < 0.01), and so were there in grade B than in grade C (P < 0.01). Among the 234 cycles in which all the transfer embryos were derived from a similar day 2 nuclear scoring, 51 cycles originated from grade A embryos (group A) and 183 from grade B (group B), with similar clinical pregnancy rates between the two groups, while the implantation rate was higher in group A than in B (P < 0.05).

CONCLUSIONDay 2 nuclear scoring can be used to predict the devel- opment and implantation potential of embryos. A combined evaluation of day 2 nuclear scoring and day 3 embryo morphology helps identify the most viable embryos and reduce the number of embryos for transfer.

Adult ; Blastomeres ; Cell Nucleus ; physiology ; Cleavage Stage, Ovum ; Embryo Implantation ; physiology ; Embryo Transfer ; statistics & numerical data ; Female ; Fertilization in Vitro ; statistics & numerical data ; Humans ; Male ; Pregnancy ; Pregnancy Outcome ; Sperm Injections, Intracytoplasmic

OBJECTIVETo investigate the influence of the rate of morphologically normal sperm (MNS) on the clinical outcomes of conventional in vitro fertilization (IVF) in patients with one retrieved oocyte.

METHODSFrom January 2013 to January 2015, a total of 256 couples with one retrieved oocyte underwent conventional IVF in our center. According to the rate of MNS, the patients were divided into two groups: MNS < 4% (134 cycles) and MNS ≥ 4% (122 cycles). We compared the rates of no transferrable embryo cycles, fertilization, cleavage, normal fertilization, abnormal fertilization, high-quality embryo and transferrable embryo between the two groups. A total of 75 fresh embryo transfer cycles were performed, 43 in the MNS < 4% group and the other 32 in the MNS ≥ 4% group. We also compared the rates of implantation, clinical pregnancy and abortion between the two groups.

RESULTSThere were no statistically significant differences between the two groups in the rates of no transferrable embryo cycles, fertilization, cleavage, normal fertilization, abnormal fertilization, high-quality embryo and transferrable embryo (P > 0.05). The rates of implantation, clinical pregnancy and abortion exhibited no remarkable differences either in the fresh embryo transfer cycles between the two groups (P > 0.05).

CONCLUSIONThe rate of MNS does not affect the clinical outcomes of conventional IVF in patients with one retrieved oocyte.

Abortion, Spontaneous ; Cleavage Stage, Ovum ; Embryo Implantation ; Female ; Fertilization ; Fertilization in Vitro ; methods ; statistics & numerical data ; Humans ; Male ; Oocyte Retrieval ; Pregnancy ; Pregnancy Rate ; Single Embryo Transfer ; statistics & numerical data ; Sperm Count ; Spermatozoa ; physiology

This study examined the effects of Bangdeyun on the expressions of nuclear factor-kappaB (NF-kappaB), interferon-gamma (IFN-gamma) and interleukin-10 (IL-10) in the endometrium of mice with embryo implantation dysfunction (EID) during the implantation time (namely on pregnancy day 5, 6, 7 and 8) and explored the local immune regulatory effects of Bangdeyun. The gestational mice were randomly divided into normal group, model group and Bangdeyun-treated group. EID models of mice were established by using indomethacin. The endometrial expression of NF-kappaB was detected by immunohistochemistry and Western blotting. IFN-gamma and IL-10 were measured by enzyme-linked immunosorbent assay (ELISA). The results showed that in the normal group, NF-kappaB and IFN-gamma were weakly expressed and IL-10 was strongly expressed in the endometrium during the whole implantation period. In the model group, the expressions of NF-kappaB and IFN-gamma were increased on pregnancy day 5, 6 and 7, and IL-10 expression decreased during the whole implantation time when compared with those in the normal group (P<0.01 for all). In the Bangdeyun-treated group, little amount of NF-kappaB and IFN-gamma was expressed and IL-10 expression was strong, much the way they were expressed in the normal group (P>0.05). The expressions of NF-kappaB and IFN-gamma were much lower in the Bangdeyun-treated group than those in the model group on pregnancy day 5, 6 and 7 (P<0.01 for all), while the expression of IL-10 was much higher than in the model group during the whole implantation time (P<0.01). It was suggested Bangderun may favor a shift from Th1- to Th2-type immune response, therefore inhibiting the maternal immune rejection, inducing the immune tolerance and improving the fetal implantation.
Drugs, Chinese Herbal/*pharmacology ; Embryo Implantation, Delayed/*drug effects ; Embryo Implantation, Delayed/immunology ; Endometrium/*immunology ; Endometrium/metabolism ; Interferon-gamma/genetics ; Interferon-gamma/metabolism ; Interleukin-10/genetics ; Interleukin-10/metabolism ; NF-kappa B/genetics ; NF-kappa B/metabolism

OBJECTIVETo observe the effect of Bushen Antai Recipe (BAR) on expression of prostaglandin I2 (PGI2) and its nuclear receptor peroxisome proliferators-activated receptor delta (PPARdelta) at implantation site in mice with blastocyst implantation dysfunction.

METHODSPregnant mice were divided into three groups randomly, the normal group, the model group and the BAR group. The pregnant uterus of all mice was cut off on the 4th (D4), 5th (D5), 6th (D6) and 8th (D8) day of pregnancy for determining the PGI2 expression with radio immunoassay; and the mRNA and protein expression of PPARdelta with RT-PCR and immunohistochemistry at implantation site.

RESULTSPGI2 expression in the model group was obviously lower than that in the normal group (P < 0.01), and also lower than that in the BAR group (P < 0.01), while the index was insignificantly different between the normal and the BAR group. Compared with the normal group, the expression of PPARdelta in the model group was delayed temporally and spatially (P < 0.05), while that in the BAR group was not significantly different.

CONCLUSIONBAR can improve the implantation in mice with blastocyst implantation dysfunction through promoting the PGI2 expression and its nuclear receptor PPARdelta at implantation site.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Embryo Implantation, Delayed ; drug effects ; Endometrium ; drug effects ; metabolism ; Epoprostenol ; biosynthesis ; Female ; Immunohistochemistry ; Male ; Mice ; PPAR delta ; biosynthesis ; genetics ; Pregnancy ; RNA, Messenger ; biosynthesis ; genetics ; Radioimmunoassay ; Reverse Transcriptase Polymerase Chain Reaction

Asoprisnil, a member of the selective progesterone receptor modulators, exerts high progesterone receptor selectivity, endometrial targeted advantages and significant anti-implantation effect in rats. The purpose of this study was to confirm the anti-implantation effect of asoprisil, investigate the ultrastructural changes of the peri-implantation endometrium in mice and explore the effect of asoprisnil on endometrial receptivity and its targeted contraceptive proficiency. Post-coitus mice were administered with different dosages (0.2, 0.1, 0.05 mg·g(-1)·day(-1)) of asoprisnil from day 1 of pregnancy to day 3. Then 3 animals in each group were killed on day 5 of pregnancy, and uteri were collected to examine the ultrastructural changes of endometria under a transmission electron microscope (TEM). A total of 80 animals were sacrificed on day 8 of pregnancy, and the uterine horns were examined for the presence or absence of nidation sites and the number of implantation embryos. The results showed that the implantation rate and the average number of implantation embryos in asoprisnil groups were statistically significantly decreased as compared with the vehicle control group (P<0.05). The TEM results revealed that, in vehicle control group, the tight junction between the luminal epithelia cells was short and straight, the gap was wide; the luminal epithelia cells were covered with plenty of short, clavate and neatly arranged microvilli; the endometril stromal cells were large with plenty of cytoplasm, and showed significant decidual change; there was more than one nucleus in stromal cells, and the karyotheca was integrity. In low dosage and high dosage asoprisnil groups, the tight junction was longer and more curve than in the vehicle control group; microvilli were uneven and asymmetrically distributed in luminal epithelia; the stromal cells were small and the decidual change was not significant; there were karyopyknosis and karyolysis in stromal cells; there were abnormal thick-wall vessels in the endometrium. It was suggested that asoprisnil changed the ultrastructure of the endometrium in implantation window, disturbed the endometrial receptivity and finally resulted in embryo implantation failure.
Animals ; Contraception, Postcoital ; methods ; Embryo Implantation, Delayed ; drug effects ; physiology ; Endometrium ; drug effects ; physiology ; ultrastructure ; Estrenes ; administration & dosage ; Female ; Mice ; Oximes ; administration & dosage ; Oxytocics ; administration & dosage ; Pregnancy ; Pregnancy, Animal ; Treatment Outcome

OBJECTIVETo explore the mechanism of Jiantai liquid on the endometrium development of embryo implantation dysfunction mice.

METHODThe model of embryo implantation dysfunction mice was induced by mifepristone and treated by Jiantai liquid. All animals were sacrificed on day 8 of pregnancy. Estradiol and progesterone concentrations in serum and endometrium tissue homogenates were measured by radioimmunoassay method, the endometial expressions of estrogen receptor (ER)and progesterone receptor (PR)assessed by immunohistochemical SP method.

RESULTThere were no significantly differences in the estradiol and progesterone concentrations in serum and uterus tissue homogenates among three groups( P > 0.05). Absorbency and area rate of ER, PR in model group' s gland and stroma were higher than those in model group(P < 0.05), which was similar with the control group( P > 0.05).

CONCLUSIONJiantai liquid increase the implantation rate and improve the endometrial development by increasing the expressions of ER, PR in endometrium of embryo implantation dysfunction

Animals ; Astragalus membranaceus ; chemistry ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Embryo Implantation, Delayed ; drug effects ; Endometrium ; metabolism ; Female ; Loranthaceae ; chemistry ; Male ; Mice ; Mifepristone ; antagonists & inhibitors ; pharmacology ; Plants, Medicinal ; chemistry ; Receptors, Estrogen ; metabolism ; Receptors, Progesterone ; metabolism ; Salvia miltiorrhiza ; chemistry