Acbp is essential for decidualization during early pregnancy in mice.

Decidualization of uterine stromal cells plays an important role in the establishment of normal pregnancy. Previous studies have demonstrated that Acyl-CoA binding protein (Acbp) is critical to cellular proliferation, differentiation, mitochondrial functions, and autophagy. The characterization and physiological function of Acbp during decidualization remain largely unknown. In the present study, we conducted the expression profile of Acbp in the endometrium of early pregnant mice. With the occurrence of decidualization, the expression of Acbp gradually increased. Similarly, Acbp expression was also strongly expressed in decidualized cells following artificial decidualization, both in vivo and in vitro. We applied the mice pseudopregnancy model to reveal that the expression of Acbp in the endometrium of early pregnant mice was not induced by embryonic signaling. Moreover, P4 significantly upregulated the expression of Acbp, whereas E2 appeared to have no regulating effect on Acbp expression in uterine stromal cells. Concurrently, we found that interfering with Acbp attenuated decidualization, and that might due to mitochondrial dysfunctions and the inhibition of fatty acid oxidation. The level of autophagy was increased after knocking down Acbp. During induced decidualization, the expression of ACBP was decreased with the treatment of rapamycin (an autophagy inducer), while increased with the addition of Chloroquine (an autophagy inhibitor). Our work suggests that Acbp plays an essential role in the proliferation and differentiation of stromal cells during decidualization through regulating mitochondrial functions, fatty acid oxidation, and autophagy.

THBS1 regulates trophoblast fusion through a CD36-dependent inhibition of cAMP, and its upregulation participates in preeclampsia.

Preeclampsia is a pregnancy complication which threatens the survival of mothers and fetuses. It originates from abnormal placentation, especially insufficient fusion of the cytotrophoblast cells to form the syncytiotrophoblast. In this study, we found that THBS1, a matricellular protein that mediates cell-to-cell and cell-to-matrix interactions, is downregulated during the fusion of primary cytotrophoblast and BeWo cells, but upregulated in the placenta of pregnancies complicated by preeclampsia. Also, THBS1 was observed to interact with CD36, a membrane signal receptor and activator of the cAMP signaling pathway, to regulate the fusion of cytotrophoblast cells. Overexpression of THBS1 inhibited the cAMP signaling pathway and reduced the BeWo cells fusion ratio, while the effects of THBS1 were abolished by a CD36-blocking antibody. Our results suggest that THBS1 signals through a CD36-mediated cAMP pathway to regulate syncytialization of the cytotrophoblast cells, and that its upregulation impairs placental formation to cause preeclampsia. Thus, THBS1 can serve as a therapeutic target regarding the mitigation of abnormal syncytialization and preeclampsia.

[Comparison of the efficiency between in-vitro maturation and in-vitro fertilization after early follicular phase GnRH agonist down-regulation in infertile women with polycystic ovary syndrome].

OBJECTIVE: To compare the outcomes of in-vitro maturation (IVM) and in-vitro fertilization (IVF) after early follicular phase gonadotropin-releasing hormone agonist (GnRH-a) down-regulation in infertile patients with polycystic ovary syndrome (PCOS). METHODS: From July 2010 to December 2012, 72 infertile patients with PCOS undergoing assisted reproductive technology treatment in the Affiliated First Hospital of Wenzhou Medical University were enrolled in this study. The patients were divided into 2 groups, which were patients with early follicular phase down-regulation IVM (36 cases) at IVM group and early follicular phase down-regulation long protocol IVF (36 cases) at IVF group. The laboratory parameters and clinical outcomes were compared between two groups. RESULTS: (1) Lab parameters: a total of 442 oocytes were retrieved in group IVM, and 560 were in group IVF. The rate of mature oocytes of 83.8% (469/560) and high-quality embryos of 70.9% (212/299) at group IVF were significantly higher than that of group IVM[54.1% (239/442) and 50.7% (73/144), retrospectively, P < 0.01]. In group IVM, the average duration of gonadotropin (Gn) was (2.8 ± 1.5) days and the average dosage of Gn was (285 ± 169) U, which were significantly lower than (11.0 ± 1.0) days and (1499 ± 165) U in group IVF (P < 0.01). The mean number of oocytes retrieved 12.8 ± 2.5, fertilization rate of 64.8% (155/239), and implantation rate of 31% (23/74) in group IVM and 15.6 ± 3.1, 65.5% (307/469), 31% (23/74) in group IVF, which did not reach statistical difference (P > 0.05) . (2) Clinical outcomes: the clinical pregnancy rate (17/31, 55%) of IVF group was not significantly higher than that 44% (14/32) at IVM group (P > 0.05). The abortion rate was 1/17 at Group IVF and 1/14 in group IVM, which did not show statistical difference. Women at IVM group has no ovarian hyper-stimulation syndrome (OHSS) cycle, group IVF has 31% (11/36) cycles presented moderate and severe OHSS. CONCLUSIONS: Infertile patients with PCOS undergoing IVM and IVF treatment after early follicular phase GnRH-a down-regulation can get satisfactory laboratory and clinical outcome. In addition to short treatment cycle, IVM can also avoid the occurrence of OHSS completely, but it has a rising trend in the abortion rate. IVF has a high incidence of OHSS, meanwhile, it increases the dosage of gonadotropins.

[Study on clinical effect on infertility women with polycystic ovary syndrome treated by in vitro maturation and in vitro fertilization-embryo transfer].

OBJECTIVE: To compare clinical and laboratory outcomes of in vitro maturation (IVM) with in vitro fertilization-embryo transfer (IVF-ET) in treatment of infertility associated with polycystic ovary syndrome (PCOS). METHODS: From Jan.2007 to Dec.2010, infertile patients with PCOS underwent 701 cycles in First Affiliated Hospital of Wenzhou Medical College were studied retrospectively. Those were divided into 293 cycles of IVM group and 408 cycles of IVF/intra-cytoplasmic sperm injection (ICSI) group. The average transplantation rate, mean number of retrieval oocytes, maturation rate, fertilization rate, cleavage rate, high quality embryo rate, embryo implantation rate, pregnancy rate per transfer, pregnancy outcomes and incidence of ovarian hyperstimulation syndrome (OHSS) of the two methods of treatment were compared between two groups. RESULTS: There were 275 cycles in IVM group and 342 cycles in IVF/ICSI group established embryo transfer. The transplantation rate was 93.9% (275/293) in IVM group and 83.8% (342/408) in IVF/ICSI, which reached statistical difference (P < 0.01). The maturation rate of 56.64%, cleavage rate of 88.08%, high quality embryo rate of 38.72% and embryo implantation rate of 17.8% in IVM group were significantly lower than 91.09%, 94.91%, 51.50% and 25.4% in IVF/ICSI group (all P < 0.01). The clinical pregnancy rate per transfer were 37.8% (104/275) in IVM group and 44.2% (151/342) in IVF/ICSI group, which did not show statistical difference (P > 0.05). The mean number of oocytes (12.9 ± 6.5 vs. 12.9 ± 7.9) and fertilization rate (76.52% vs. 70.75%) didn't show significant difference between IVM group and IVF/ICSI group (P > 0.05). The 21.3% (87/408) cycles presented mild to moderate OHSS and 2.0% cycles (8/408) presented severe OHSS in IVF/ICSI group. While, no OHSS cycles were observed in IVM group. CONCLUSION: IVM could get similar clinical pregnancy rates compared with IVF/ICSI in patient with PCOS, however, it can avoid occurrence of OHSS.

[Clinical study on in vitro maturation of immature oocytes transferred from conventional in vitro fertilization.].

OBJECTIVE: To investigate clinical effect of in vitro maturation (IVM) of immature oocytes transferred from conventional in vitro fertilization embryo transfer (IVF-ET) cycles. METHODS: From January 2008 to June 2009, medical documents of 155 infertile patients underwent IVF-ET in the Reproductive Medical Center of First Affiliated Hospital of Wenzhou Medical College were analyzed retrospectively. If more than 20 oocytes were monitored after 5 - 7 days of ovulation induction or follicular developmental retardation were confirmed after 8 - 13 days of ovulation induction, according to patients' wish, IVM were transferred in 60 cycles (group A). In the mean time, IVF was continued in 95 cycles (group B). The mean dosage of gonadotropin, the cancellation rate of cycles, the mean numbers of oocytes retrieved and maturation, the rate of fertilization and excellent quality embryos, pregnancy outcome and the incidence rate of ovarian hyperstimulation syndrome (OHSS) were compared and analyzed. RESULTS: The rates of embryo transfer were 92% (55/60) in group A and 63% (60/95) in group B, which showed significant differences (P < 0.05). In group A, the mean dosage of the gonadotropin, the mean number of oocytes retrieved, the cleavage rate and OHSS rate were (1030 +/- 468) U, 10 +/- 6, 82.2% (231/281) and 0, and were (1544 +/- 338) U, 14 +/- 4, 94.0% (502/534) and 35% (21/60) in group B, respectively, all data above exhibited statistical difference between two groups (P < 0.05). However, the rates of fertilization and excellent quality embryos had no significant differences between two groups (P > 0.05). In group A, the rate of clinical pregnancy per transfer was 53% (29/55) and multiple pregnancy was 14% (4/29), and were 47% (28/60) and 32% (9/28) in group B, they all had no significant differences (P > 0.05). CONCLUSION: IVM of immature oocytes used in conventional IVF cycles not only obtained a high clinical pregnancy rate, but also reduced gonadotropin using dosage and avoided OHSS completely.

[A study on bisulfite sequencing method for methylation status of imprinted genes in single human oocytes].

OBJECTIVE: To establish and improve the method of bisulfite sequencing for methylation status of imprinted genes in single human oocytes. METHODS: Single superovulated immature human oocyte was embedded into low melting point agarose, followed by bisulfite treatment and polymerase chain reaction (PCR) amplification of the H19 and MEST genes. The PCR products were then subjected to TA cloning and sequencing to determine the methylation status. RESULTS: With the modified methods of embedding and bisulfite treatment, we achieved a high PCR success rate of 82.46%, with the somatic cell contamination rate as low as 7.14%. The sequencing results showed no non-CpG cytosine and exact conformity to the theoretical sequences. CONCLUSION: The bisulfite sequencing method we used to determine the methylation status of imprinted genes at the single-cell level was highly efficient and reliable, which can serve as a foundation for the further study of the influences of human assisted reproductive technology on genomic imprinting.

Establishment and characterization of two embryonic cell lines of Bombyx mori.

Two cell lines, i.e., BmE-SWU1 and BmE-SWU2, were established from silkworm embryonic tissues of the reversion phase through primary culture in Grace's medium supplemented with 20% fetal bovine serum. The BmE-SWU1 cell line mainly included diploid spindle cells and round cells, which were large and had severe heteroploidy karyotypes. The population doubling time of the 30th passage of the cell line was 58.7 hr. BmE-SWU2 cells were oblong or round, and small. The population doubling time for the 30th passage of the cell line was 46.6 hr. Of BmE-SWU2 cells 89.9% were diploid (2n=56). Both strains were attached to epithelial-like cell lines and were susceptible to Bombyx mori nucleopolyhedroviruse (BmNPV). Inter simple sequence repeat (ISSR) fingerprinting of silkworm embryonic cell line was obtained.

[Meiotic segregation results of male reciprocal chromosome translocations].

OBJECTIVE: To analyze the meiotic segregation results of male reciprocal chromosome translocation by fluorescence in situ hybridization (FISH). METHODS: Multi-color FISH using 3 combined probes located in any 3 chromosome segments on both sides of two breakpoints was performed on the de-condensed sperm head to analyze the sperm chromosomal contents and segregation patterns. RESULTS: Four male reciprocal translocation carriers were included in the study, with the karyotypes of 46, XY, t(2;18) (p16; q23); 46, XY, t(4;6) (q34;q21); 46, XY, t(8;13) (q23;q21) and 46, XY, t(4;5) (4q31;5q13), respectively. The results showed that 4 carriers had different proportions of various segregated spermatozoa. The spermatozoa of alternate, adjacent-1, adjacent-2, 3:1, non-disjunction in meiosis II, and 4:0 or diploidy accounted for 27.1%-49.4%, 26.9%-37.6%, 2.7%-15.7%, 8.6%-32.7%, 0.2%-1.9%, and 0.1%-0.4%, respectively. CONCLUSION: For each-reciprocal translocation carrier seems to have a particular meiotic segregation results, FISH analysis on sperm head should be done for each carrier in order to provide an accurate genetic counseling.