[Application of single-sperm sequencing in resolving the carrier status of preimplantation genetic testing for chromosomal structural rearrangements in Robertsonian translocations].

OBJECTIVE: To investigate the application value of single-sperm sequencing in resolving the carrier status of preimplantation genetic testing (PGT) for chromosomal structural rearrangements in Robertsonian translocations. METHODS: Haplotypes were constructed by single-sperm isolation combined with single-sperm sequencing for a patient with 45, XY, der(13; 14)(q10; q10). Twenty single-sperm samples were isolated by mechanical braking and subjected to whole-genome amplification (WGA), and then the Asian Screening Array (ASA) gene chip was used to detect the 183 708 single nucleotide polymorphisms (SNP) of the WGA products. The single sperm associated with the translocation that could be used as haplotype inference was detected by copy number variation (CNV) sequencing, and the chromosomal haplotypes with normal and Robertsonian translocations were inferred. Three biopsy samples of embryonic trophoblast cells were used as the objects. After whole-genome amplification, high-throughput sequencing was employed to determine the status of the translocation chromosome carried by the embryos. The available blastocysts were selected for transfer, and the amniotic fluid samples were taken at 18 weeks of gestation to confirm whether the fetus carried the pathogenic mutation. RESULTS: A total of 6 037 SNP sites were screened by single-sperm sequencing, and 30 sites selected to distinguish normal and translocation haplotypes. Preimplantation haplotype analysis showed that all the three embryos were euploids without Robertsonian translocation chromosome. Genetic testing of amniotic fluid in the second trimester confirmed that the karyotype of the fetus was 46, XN, carrying no Robertsonian translocation chromosome. CONCLUSION: For male carriers of Robertsonian translocation, single sperm sequencing can be used to screen SNP sites to construct haplotypes for distinguishing normal and Robertsonian translocation embryos, and to provide a basis for embryo selection by preimplantation chromosomal structural genetic testing.

Expression and regulation of androgen receptor in the mouse uterus during early pregnancy and decidualization.

The androgen receptor (AR) is a ligand-activated transcription factor that is important for both the male and female reproductive systems. The expression and regulation of AR in the uterine endometrium during early pregnancy and decidualization remain relatively under-investigated, so we sought to immunohistochemically examine the spatiotemporal expression of AR in mouse uteri during the peri-implantation period as well as in response to specific steroid hormones. AR protein was found in the nuclei of uterine stromal cells starting on pregnancy Days 1 and 2, with its abundance increasing on Days 3 and 4. From pregnancy Days 5 to 9, however, the expression of AR markedly declined in stromal zones of uteri. No signal was detected in the decidualized cells surrounding the site of embryo implantation; moreover, no AR immunostaining was observed in decidualized uterine cells in an artificial oil-induced model of decidualization. Progesterone significantly inhibited AR protein expression, whereas estrogen dramatically elevated AR abundance in the stroma of ovariectomized mouse uteri. Taken together, our results are the first to demonstrate that decidualization and progesterone significantly inhibited the AR protein expression in vivo, whereas estrogen increased AR protein levels in the stromal cells of mouse uteri. These responses might be advantageous for the proliferation and differentiation of uterine stroma and for embryo implantation during early pregnancy.

Exposure to di(2-ethylhexyl) phthalate inhibits luteal function via dysregulation of CD31 and prostaglandin F2alpha in pregnant mice.

BACKGROUND: Di(2-ethylhexyl) phthalate (DEHP) exposure reduces embryo implantations, increases embryonic loss, and decreases fetal body weights. However, whether it is associated with the alteration of luteal function remains unknown. Thus, our aim in this study was to explore the effect and mechanism of DEHP on luteal function in pregnant mice in vivo. METHODS: Mice were administered DEHP by gavage at 125, 250, 500 mg/kg/day from gestational days (GD) 1 to 9 or 13. Levels of serum progesterone and estradiol were measured by radioimmunoassay. The numbers and sizes of corpora lutea were calculated by ovarian histomorphology. Steroidogenic enzymes were assessed by qRT-PCR. CD31 protein was detected by immunocytochemistry, and prostaglandin F2alpha (PGF2alpha) levels were evaluated by enzyme immunoassay. RESULTS: Treatment with DEHP significantly inhibited progesterone secretion in pregnant mice in a dose-dependent manner but did not inhibit estradiol production on GD 9 and 13. Treatment also showed concomitant decreases in transcript levels for key steroidogenic enzymes (CYP11A, 3ß-HSD, and StAR) on GD 13. Furthermore, DEHP administration significantly reduced the numbers and sizes of corpora lutea on GD 13. No significant changes in the ratio of ovary weight vs. body weight were observed between the control group and treated animals on GD 9 and 13. In addition, treatment with DEHP significantly inhibited CD31 expression of corpora lutea, whereas plasma PGF2alpha levels in DEHP treatment groups were significantly higher compared with the control groups on GD 9 and 13. CONCLUSIONS: The results show DEHP significantly inhibits luteal function of pregnant mice in vivo, with a mechanism that seems to involve the down-regulation of progesterone and steroidogenic enzymes message RNA, the decrease in CD31 expression, and the increase in PGF2alpha secretion.

Actinidia macrosperma C. F. Liang (a Wild Kiwi): Preliminary Study of Its Antioxidant and Cytotoxic Activities.

The antioxidant potential of Actinidia macrosperma C. F. Liang (Actinidiaceae) was investigated in vitro for total phenolic content, along with total antioxidant activity (TAA), 1,1-diphenyl 2-picryl hydrazyl (DPPH), and lipid peroxidation (LP). The results indicated that different polarity extracts of A. macrosperma exhibit different biological activities, which depends mainly on the presence of phenolic compounds. The antioxidant activity was in the following decreasing order: MeOH extract > EtOAc extract > aqueous extract > CHCl(3) extract > Hexane extract. Moreover, the cytotoxic activity of this plant by MTT dye assay using SMMC-7721 has been determined also. The hexane, EtOAc, and CHCl(3) extracts showed cytotoxicity in a dose-dependent manner. Methanol and aqueous extracts, however, showed weak activities in this test. And a very significant cytotoxic activity, not significantly different from the positive control of quercetin, was observed in CHCl(3) extract.

Reduced expression of follicle stimulating hormone receptor mRNA and protein in pregnancies complicated by pre­eclampsia.

Expression and function of the follicle­stimulating hormone receptor (FSHR) are traditionally thought to be limited to the ovary in females. However, recent studies have indicated that the FSHR is also expressed in endothelial cells of the placental vasculature, and that the haploinsufficiency of the feto­placental FSHR impaired the growth of the mouse placenta. The aim of the current study was to investigate the placental expression of FSH and FSHR in pregnancies complicated by pre­eclampsia. Placental tissue samples were collected from 20 pregnancies with pre­eclampsia and 25 normal pregnancies. Placental FSH and FSHR mRNA levels were measured by reverse transcription­quantitative polymerase chain reaction (RT­qPCR), while FSH, FSHR and CD31 protein expression were examined by immunohistochemistry. Additionally, levels of serum FSH were measured by chemical luminescence immunity assay. The results demonstrated that no significant difference was observed in serum FSH levels and expression levels of placental FSH mRNA and protein between normal pregnancy and pre­eclampsia. However, RT­qPCR results indicated that the expression level of FSHR mRNA in pre­eclamptic placental samples was significantly lower than normal pregnancies. Immunostaining results from normal pregnant samples indicated that the FSHR protein was strongly expressed in the endothelial cells of blood vessels in the chorionic villi, moderately expressed in stromal cells of the villus, but not expressed in trophoblast cells of the term placenta. The staining intensity of FSHR­positive area was significantly lower in the placental villi of pre­eclampsia, when compared with the normal control group. In conclusion, expression levels of placental FSHR mRNA and protein are significantly reduced in pregnancies complicated with pre­eclampsia in the present study. Further studies may investigate whether FSHR could be used as a biomarker for the prediction of pre­eclampsia.

Maternal exposure to di-(2-ethylhexyl) phthalate disrupts placental growth and development in pregnant mice.

Di-(2-ethylhexyl) phthalate (DEHP) is used as a plasticizer and widely dispersed in the environment. DEHP exposure reduces embryo implantations, increases embryonic loss, and decreases fetal body weights. However, no detailed information is available about the effect of DEHP on the placentation during pregnancy. Thus, our aim was to explore the effect of DEHP on the growth and development of placenta in vivo. Mice were administered DEHP by gavages at 125, 250, 500 mg/kg/day from gestational days (GD) 1 until sacrifice. Results showed that DEHP treatment significantly reduced the weight of placenta at GD 13. Histopathologically, in DEHP-treated group, the ectoplacental cones significantly became smaller at GD9, and total area of placenta and area of spongiotrophoblast were significantly reduced at GD 13. Expression levels of Ascl2, Esx1 and Fosl1 mRNA dramatically decreased in DEHP-treated placenta at GD 13. DEHP administration disrupted labyrinth vascularization of placentas, and inhibited proliferation and induced apoptosis of placenta by the activation of caspase-3 and -8, up-regulation of Bax and down-regulation of Bcl-2 mRNA and protein at GD 13. In conclusion, these results suggest that adverse pregnancy outcomes including low birth-weight and pregnancy loss exposed to DEHP are possibly mediated, at least in part, via the suppression of placental growth and development.

[The effects of human cytomegalovirus infection on college students' neurobehaviors].

OBJECTIVE: To explore relevant between human cytomegalovirus (HCMV) infection and college students' neurobehaviors. METHODS: 87 college students were enlisted. They were tested with Bole. Neurobehavioral evaluation system (B. NES), and HCMV IgG antibody was detected after separation of serum. We analyzed the test results of B. NES by SPSS software. RESULTS: 76 college students were infected by HCMV in the past and 11 college students were not infected. The infected group scored 8.89 +/- 6.60 in depression aspect of emotion state test, while control group got 15.73 +/- 9.00. There was Significant difference between infection group and control (P < 0.05). There were no significant differences in other aspects of emotion states, study and memory, perception and mental movement (P > 0.05). CONCLUSION: HCMV infection is associated with depression status.