After implantation, complex and highly specialized molecular events render functionally distinct organ formation, whereas how the epigenome shapes organ-specific development remains to be fully elucidated. Here, nano-hmC-Seal, RNA bisulfite sequencing (RNA-BisSeq), and RNA sequencing (RNA-Seq) were performed, and the first multilayer landscapes of DNA 5-hydroxymethylcytosine (5hmC) and RNA 5-methylcytosine (m⁵C) epigenomes were obtained in the heart, kidney, liver, and lung of the human foetuses at 13-28 weeks with 123 samples in total. We identified 70,091 and 503 organ- and stage-specific differentially hydroxymethylated regions (DhMRs) and m⁵C-modified mRNAs, respectively. The key transcription factors (TFs), T-box transcription factor 20 (TBX20), paired box 8 (PAX8), krueppel-like factor 1 (KLF1), transcription factor 21 (TCF21), and CCAAT enhancer binding protein beta (CEBPB), specifically contribute to the formation of distinct organs at different stages. Additionally, 5hmC-enriched Alu elements may participate in the regulation of expression of TF-targeted genes. Our integrated studies reveal a putative essential link between DNA modification and RNA methylation, and illustrate the epigenetic maps during human foetal organogenesis, which provide a foundation for for an in-depth understanding of the epigenetic mechanisms underlying early development and birth defects.
Humans ; DNA Methylation ; RNA ; Epigenesis, Genetic ; DNA/genetics* ; Basic Helix-Loop-Helix Transcription Factors/genetics*

Objective:In vitro fertilization and embryo transplantation in patients with endometrial implantation window phase of natural killer cell ( uNK ) influence on the endometrial microvascular and uterine artery pulsatility index and embryo implantation.Methods:A retrospective analysis of our hospital from January 2009 to July 2013 the implementation of in vitro fertilization and embryo transfer(IVF-ET) clinical data of 37 patients,according to whether the success of pregnancy and at least 2 times for IVF-ET failure into success group(26 cases)and the failure group(11 cases),compared two groups of research object during the window of implantation in the uterus endometrial uNK cell marker CD56+, uterine artery pulsatility index, vascular endothelial marker F8 factor,alpha smooth muscle actin( alpha SMA) and myosin heavy chain( SMM) and the relationship between the expression of difference.Results:Success group based FSH 3.52±0.68(mU/ml),basal antral follicles of 9.44±2.53 compared with the failure group differences were not statistically significant(P>0.05).The successful group of uNK cells of 27.18±5.94(%),MVD 6.79±1.74 (%) ,αSMA 33.72 ±4.19 (%) , SMM 25.19 ±5.83 (%) were significantly higher than the expression rate of IVF-ET assisted reproductive failure group(P<0.05).The number of uNK cells,the proportion of endometrium during the window of implantation and MVD of 37 patients withαSMA and SMM coloring cell number proportion were positive correlation significantly( r=0.472,P=0.000<0.005),(r=0.426 ,P=0.000<0.005),(r=0.512,P=0.000<0.05).Success group during the window of implantation in the uterine artery PI=0.94±0.28,PI=0.87±0.29 failure group,differences between the two groups was not statistically significant(t=0.688,P=0.493>0.05).Conclusion:During the window of implantation in the endometrium of natural killer cells and endometrial angiogenesis was significantly associated with successful pregnancy, may have a certain relationship;uterine artery PI and pregnancy outcome successful has no obviously relationshhip.

Objective To investigate ectopic pregnancy from embryo transfer (ET)of in-vitro fertilization (IVF) cycle and intracytoplasmic sperm iujection (ICSI) cycle and frozen-thawed (FET) cycle.Methods From Jan.2005 to Dec.2010,a total of 9037 IVF-ET or ICSI-ET cycles and 4034 FET cycles were performed in our reproductive medicine center,Affiliated First Hospital of Zhengzhou University.The incidence of ectopic pregnancy rate was studied in fresh cycles IVF-ET (5998) and ICSI-ET (3039) cycles,and natural FET (2198) and hormone replacement (E-P) FET (1836) cycles.ResultsOf 4034 FET cycles,1090 clinical pregnancies and 26 ectopic pregnancies were observed,the incidence of ectopic pregnancy was 2.38％ (26/1090).Of 9037 fresh cycles,3602 cycles were clinical pregnancy,and 133 cycles were ectopic pregnancy,and the incidence of ectopic pregnancy was 3.69％ (133/3602).The ectopic pregnancy rate in FET cycles was lower than in fresh cycles significantly (P ＜0.05).Of 3039 fresh ICSI-ET cycles,the incidence of ectopic pregnancy was 2.62％ ( 34/1298 ) in 1298 clinical pregnancies.Of 5998 IVF-ET cycles,2304 clinical pregnancies were observed,the incidence of ectopic pregnancy was 4.30％ (99/2304).Ectopic pregnancy rate in the fresh ICSI-ET cycles was lower than that of IVF-ET group significantly (P ＜ 0.01 ).The ectopic pregnancy rate in the natural FET cycles was 1.46％ (8/547),which was significantly lower than 3.31％ ( 18/543 ) in E-P group ( P ＜ 0.05 ).ConclusionsThe incidence of ectopic pregnancy of FET cycles was significantly lower than that of fresh embryo transfer cycles.The application of exogenous sex hormones in assisted reproductive cycles might increase occurrence of ectopic pregnancy.

Objective To evaluate the effects on pregnancy outcome of freezing time from oocyte retrieval and thawing method for metaphase Ⅱ human oocytes vitrification. Methods From Mar 2007 to Mar 2009, the clinical outcome of 30 infertile women undergoing vitrified-thawing oocytes of in vitro fertilizationembryo transfer(IVF-ET) in the Reproductive Medical Center of the First Affiliated Hospital of Zhengzhou University was studied retrospectively, including 21 women with double fallopian tube obstruction and 9 women's husband azoospermia. All infertile women were divided into three groups, including 5 cases in group A (freezing between 4 and 5 hours from oocyte retrieval and conventional thawing method), 9 cases in group B (freezing within 2 hours from retrieval and conventional thawing method) and 16 cases in group C (freezing within 2 hours from retrieval and improved thawing method). The vitrified oocytes were preserved for 2 months to I year and thawed for Intracytoplasmic sperm injection (ICSI) and embryo transfer. The outcome of IVF and pregnancy were recorded. Results (1) The rates of oocyte survival was (65±33) % in group B and (72±23)% in group C and the rate of transfer cycle was 9/9 in group B and 16/16 in group C, which were all significantly higher than (16±17) % of oocyte survival and 1/5 of transfer cycle in group A (P = 0. 001,0. 021). However, the rate of oocyte survival and transfer cycle between group B and group C did not reach statistical difference (P > 0. 05). The rate of implantation and clinical pregnancy of (33±38) % and 9/16 in group C were significantly higher (4±11)% and 1/9 in group B (P =0. 033,0. 040).(2)The mean age of women in group C were (28.6±2.1) in oneself oocyte, (28.0±4.6) in donor oocyte and (28.1±3.4) in donor sperm. The rate of oocyte survival was (73±25) %, (88±10) % and (66±25) %. The rate of fertilization rate was (84. 6±0. 9) %, (79. 3±2. 0) % and (82. 8±15.0) %. The rate of implantation was (20. 0±44. 7) %, (33. 0±0. 1) % , (41.6±41.7) %. The rate of clinical pregnancy was 1/5 in oneself cycles,3/3 in donor oocyte cycles, 5/8 banked donor sperm cycles in group C. All above clinical parameters were not statistically different (P >0. 05). (3) In group A, one women underwent IVFET and no clinical pregnancy was observed. One women pregnancy was terminated at two months in group B.The clinical pregnancies rate of group C was 9/16, late abortion occurred in 1 woman, the other 8 women underwent term pregnancy, including 5 male infants and 4 female infants. All of infants showed normal Karyotype. Live-birth rates per warmed oocyte was 5.9% (8/135). The mean gestational weeks and birth weight of the infants were (39. 4±0. 9) weeks and (3574±569) g, respectively. Conclusions Embryo quality and clinical outcome of thawing cycles could be significantly improved when oocyte vitrification was performed within 2 hours from oocyte retrieval and improved thawing method.

Objective To compare the advantages of three-dimensional(3D) and two-dimensional(2D) ultrasound in embryo transfer. Methods A total of 319 patients accepted embryo transfter were included in this study. 2D and 3D ultrasound were used to investigate the uterine cavity and transfer distance from the fundus (TDF),respectivly. They were divided into four groups according to TDF difference(D-TDF) between 2D and 3D ultrasound(group of DTDF<3mm,group of DTDF3～5mm,group of 6～9 mm,group of DTDF≥10 mm. Pregnancy outcomes among the four groups were compared. Results Of the 319 patients, 41 were observed to have abnormal uterine cavity. For 140 patients, the TDF measured by 2D ultrasound were different from that measured by 3D ultrasound. Clinical pregnancy rate and implantation rate were found lowest in group of TDF≥10 mm mm (7.7% vs 34.1%,38.1% ,40.0% and 3.6% vs 18.2% ,21.2% ,20.0%, P <0.05). Conclusions 2D ultrasound is limit and deficient for embryo transfer, especially for the visualization of uterine cavity and location of catheter tip, however, it may be better achieved with 3D ultrasound. It is helpful to use the 3D ultrasound to place the catheter tip accurately and improve the pregnancy rate of embryo transfer.

Objective To investigate the incidence of and clinical factors influencing neonatal birth defects from different assisted reproductive technology. Methods Between October 1998 and December 2006,1271 newborns from mothers treated by in vitro fertilization techniques [ including in vitro fertilization (IVF), intracytoplasmic sperm injection (1CSI) and thaw embryo transfer (Thaw-ET) ] matched with 269 newborns from mothers treated by artificial insemination were enrolled in Reproductive Medicine Center in First Hospital Affiliated to Zhengzhou University. Their medical information was analyzed retrospectively to compared neonatal characteristics, the incidence of birth defect and anomalous organs involved between in vitro fertilization group and artificial insemination group. Results In group of in vitro fertilization, those newborns with low birth weight from IVF, ICSI and Thaw-ET were 20. 0% ( 134/671 ), 22. 4% (92/410), 18.9% (36/190)respectively, which were more than 11.5% (31/269) cases in group of artifical semination with statistical significance (P < 0. 05 ). The rates of multiple pregnancy of 23.8% ( 160/671 ), 25.4% (104/410) ,21.1% (40/190) in subgroup of 1VF, ICSI and Thaw-ET were significantly higher than 10. 0% ( 27/269 ) in group of artifical insemination( P < 0. 05 ). The rate of macrosomia in group of in vitro fertilization was significantly lower than that of artificial insemination group (3.9% vs 8. 2%, P <0.05). However, the incidence of birth defect involved in various organs did not show significant difference between two groups ( P>0.05 ). Conclusions The incidence of multiple pregnancy demonstrated obviously increasing trends born with various In Vitro Fertilization techniques, which pave the way to high risk pregnancy. However, the incidence of newborn birth defect was not increased significantly. Thus, to lower occurrence of multiple pregnancy was the key approach to obtain neonates health.

0.05).The effective rate on relieving traditional Chinese medical syndrome was 95.0% in the treatment group and 73.3% in the control group,the difference being significant(P0.05).After treatment,the parameters of 24-h dynamic blood pressure were improved in both groups(P