Sperm-specific protein ACTL7A as a biomarker for fertilization outcomes of assisted reproductive technology / 亚洲男科学杂志(英文版)

Obtaining high-quality embryos is one of the key factors to improve the clinical pregnancy rate of assisted reproductive technologies (ART). So far, the clinical evaluation of embryo quality depends on embryo morphology. However, the clinical pregnancy rate is still low. Therefore, new indicators are needed to further improve the evaluation of embryo quality. Several studies have shown that the decrease of sperm-specific protein actin-like 7A (ACTL7A) leaded to low fertilization rate, poor embryo development, and even infertility. The aim of this study was to study whether the different expression levels of ACTL7A on sperm can be used as a biomarker for predicting embryo quality. In this study, excluding the factors of severe female infertility, a total of 281 sperm samples were collected to compare the ACTL7A expression levels of sperms with high and low effective embryo rates and analyze the correlation between protein levels and in-vitro fertilization (IVF) laboratory outcomes. Our results indicated that the ACTL7A levels were significantly reduced in sperm samples presenting poor embryo quality. Furthermore, the protein levels showed a significant correlation with fertilization outcomes of ART. ACTL7A has the potential to be a biomarker for predicting success rate of fertilization and effective embryo and the possibility of embryo arrest. In conclusion, sperm-specific protein ACTL7A has a strong correlation with IVF laboratory outcomes and plays important roles in fertilization and embryo development.

Modified strict sperm morphology threshold aids in the clinical selection of conventional in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) / 亚洲男科学杂志(英文版)

For infertility treatment, the selection of in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) is decided by multiplying indicators (including fallopian tube factors, semen count, and semen motility), except for sperm morphology. In this study, we conducted a retrospective analysis, from implantation to birth, over a period of 5 years. A total of 1873 couples with primary or secondary fallopian tube factors and an increased defective sperm morphology rate (DSMR) were divided into different groups to receive IVF or ICSI cycles. By comparing the outcomes, we found that the F1 group (DSMR <96%, IVF group 1) had higher cleavage rate, biochemical pregnancy rate, clinical pregnancy rate, and live birth rate than the F3 group (DSMR >98%, IVF group 3; P < 0.05). In contrast, there was no significant difference in the ICSI subgroups. Furthermore, a comparison of the outcomes between IVF and ICSI showed that the S3 group (DSMR >98%, ICSI group 3) had higher cleavage rate (P < 0.001), biochemical pregnancy rate (P < 0.05), clinical pregnancy rate (P < 0.05) and live birth rate (P < 0.05) than the F3 group. However, the ICSI subgroup had a lower two pronuclei fertilization rate than the IVF subgroup (P < 0.05). Our data suggest that the sperm morphology should also be considered when selecting IVF or ICSI combined with other semen parameters before the first assisted reproductive technologies (ART) cycle, especially for males with severe sperm defects.

Advances in the construction of a scaffold for cardiac tissue engineering containing homogeneous and stable carbon nanotubes / 中国组织工程研究

BACKGROUND: How to efficiently and uniformly disperse carbon nanotubes (CNTs) into a tissue-engineered scaffold is crucial to construct an ideal CNTs-Polymer composite scaffold and it is also a hotspot of research in the tissue engineering. OBJECTIVE: To review the advances in the methodology progress of constructing a scaffold for cardiac tissue engineering, which contains uniformly and stably dispersed CNTs. METHODS: The Web of Science Core Collection and PubMed were searched by the first author for related papers about CNTs dispersion in the cardiac tissue engineering published from October 2004 to January 2017. The key words were "carbon nanotubes, dispersion, cardiac tissue engineering" in English. Original research papers were searched, which were screened through titles, abstracts and contents, and then reviewed. RESULTS AND CONCLUSION: CNTs are easy to aggregate because of high surface area, high aspect ratio and rough surface. Thus, it is one of the key points to construct an ideal CNTs-Polymer composite scaffold that whether CNTs could be uniformly and stably dispersed in polymer scaffolds. In the cardiac tissue engineering, covalent or non-covalent surface modification of CNTs significantly enhances the uniformity and stability of CNTs in the polymer scaffolds, which is conducive to construct the uniformly and stably CNTs-dispersed scaffold for cardiac tissue engineering, leading to notable improvement in mechanical and electrical properties of engineered cardiac tissues.

Epidemiological Survey and Risk Factor Analysis of Recurrent Spontaneous Miscarriages in Infertile Women at Large Infertility Centers / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>A higher frequency of spontaneous miscarriage has been observed in infertile couples, and there is a higher prevalence of infertility among patients with a history of recurrent spontaneous miscarriages (RSMs; ≥2 miscarriages). This study aimed to determine the proportion of infertile patients with RSM and examine risk factors associated in patients with RSM being treated with assisted reproductive technologies.</p><p><b>METHODS</b>This cross-sectional observational study was conducted at six reproductive medicine centers in three cities of China. Data of 751 patients with at least one spontaneous miscarriage were analyzed. Demographic data and etiological factors associated with infertility were compiled and compared between patients with a single spontaneous miscarriage (SSM) and those with RSM.</p><p><b>RESULTS</b>Two hundred (26.6%, 95% confidence interval [CI]: 23.50-29.95%) patients experienced RSMs and 551 (73.4%) had a single miscarriage. The odds of RSM increased with increasing age (odds ratio [OR] = 1.06), uterine disorders (OR = 2.09), endocrine disorders (OR = 2.48), and immune disorders (OR = 2.98). Higher education level, masters or above, and a pelvic cavity disorder were associated with lower risk of RSM (OR = 0.27 and 0.46, respectively). Late spontaneous miscarriages were more frequent in patients with RSM than in those with a SSM (31.5% vs. 14.2%, respectively, P< 0.001) and were associated with a history of uterine cavity procedures (OR = 2.095) and cervical factors related to infertility (OR = 4.136, 95% CI: 1.012-16.90).</p><p><b>CONCLUSIONS</b>Compared to patients with only a SSM, the conditions of patients with RSM are more complicated. To increase the success rate of assisted reproductive technology, factors including uterus cavity adhesion, cervical relaxation, endocrine disorders, and immune disorders should be treated before assisted reproduction is initiated. These data may provide treatment guidance for infertile patients with a history of RSM.</p>

Maternal Genistein Intake Can Reduce Body Weight in Male Offspring / 生物医学与环境科学（英文）

The study objectives were to investigate the relationship between early exposure to genistein and obesity in young adulthood and to evaluate changes in reproductive health during puberty and adulthood following in utero exposure to genistein. Thirty-two female rats were randomized into four groups; low dose 400 mg genistein/kg diet group (LG), mid-dose 1200 mg genistein/kg diet group (MG), high dose 3600 mg genistein/kg diet group (HG), and control group without genistein diet (CON). Rats were fed genistein at the beginning of pregnancy along with a high-fat diet. Pups were sacrificed at week 4 and week 8 after birth. High performance liquid chromatography (HPLC) results showed a correlation between maternal genistein intake and genistein concentration in pups' plasma. Compared to CON, body weight reduced significantly in male HG group at week 8. No statistical differences were found in plasma estradiol (E2), testosterone (T), interleukin (IL)-6, and C-reactive protein (CRP) levels with early genistein exposure. Furthermore, uterine histopathology showed notable changes in groups HG and MG compared with CON at week 4 and week 8. In conclusion, maternal genistein supplement could reduce body weight in male pups and alter uterine histopathology in female pups.

Acquirement and evaluation of murine ventricular extracellular matrix / 生理学报

Cardiac extracellular matrix (ECM), generated from the process of decellularization, has been widely considered as an ideal source of biological scaffolds. However, current ECM preparations are generally difficult to be applied to generate cardiac tissue. Our research was aimed to improve decellularization protocols to prepare cardiac ECM slices. Adult murine ventricular tissues were embedded in low melting agarose and cut into 300 μm slices, and then were divided randomly into three groups: normal cardiac tissue, SDS treated group (0.1% SDS) and SDS+Triton X-100 treated group (0.1% SDS+0.5% Triton X-100). Total RNA content and protein content quantification, HE staining and immunostaining were used to evaluate the removal of cell components and preservation of vital ECM components. Furthermore, murine embryonic stem cell-derived cardiomyocytes (mES-CMs) and mouse embryonic fibroblasts (MEFs) were co-cultured with ECM slices to evaluate biocompatibility. The relative residual RNA and protein contents of ECM slices significantly decreased after decellularization. HE staining showed that SDS+Triton X-100 treatment better destroyed cellular structure and removed nuclei of ECM slices, compared with SDS treatment. Immunostaining showed that collagen IV and laminin were better preserved and presented better similarity to original cardiac tissue in ECM slices acquired by SDS+Triton X-100 treatment. However, collagen IV and laminin were significantly decreased and arranged disorderly in SDS treated group. We observed effective survival (≥ 12 days) of MEFs and mES-CMs on ECM slices acquired by SDS+Triton X-100 treatment, and signs of integration, whereas those signs were not found in SDS treated group. We concluded that, compared with traditional SDS method, new combined protocol (SDS+Triton X-100) generated ECM slices with better component and structural preservation, as well as better biocompatibility.

Efficiency of multi-round fluorescence in situ hybridization and its influencing factors in preimplantation genetic diagnosis / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the efficiency of multi-round fluorescence in situ hybridization (FISH) and its influencing factors in preimplantation genetic diagnosis (PGD).</p><p><b>METHODS</b>A total of 48 couples accepted PGD because of various reasons: 24 with Robertsonian translocations, 16 with reciprocal translocations, 2 with pericentric inversions, one with advanced maternal age who had a previous liveborn of Down syndrome, 3 suffered from sex chromosome abnormalities and 2 repeated spontaneous miscarriages. After 72 retrieval cycles, 432 cleavage stage embryos with more than six cells were biopsied on day three. Only intact nuclei (396) were hybridized in order to verify the chromosomal status of the individual embryos. If previous FISH has failed to give conclusive results while the nuclei remained undamaged, the nuclei were hybridized once again. A total of 870 times of hybridization were conducted to 396 nuclei. Signal identification rates of each round as well as the influence of different probes to the hybridization efficiency were compared. Factors leading to inconclusive FISH results were analyzed as well.</p><p><b>RESULTS</b>Five hundred and thirty five out of 870 hybridizations gave identifiable signals (61.5%). The second and third round FISH showed the best signals with an identification rate of 71.8% and 77.4%, respectively, which were significantly higher than those of the first round (52.8%, P < 0.01), the fourth round (55.8%, P < 0.05, P < 0.01), the fifth round (54.5%, P < 0.05) and the sixth round (27.3%, P < 0.01). The identification rate of centromere specific probe signals (CEP group) was 80.3% and the former three rounds in this group got the best quality of signals with an identification rate of 85.7%, 85.1% and 88.0%, respectively, which was significantly higher than that of the latter three rounds. The identification rate of other probe was much lower than with the CEP probe (55.2% vs. 80.3%, P < 0.01) and the best quality of signal in this group was achieved in the fifth round (72.7%), followed by the second round (66.1%) and the third round (63.8%). The identification rate of the first round (50.3%) and the sixth round (22.2%) were significantly lower compared with the second round (P < 0.01). During the 6 rounds of FISH, 335 hybridizations did not give conclusion results (38.5%, 335/870). The main cause of unidentification was weak signals (20.9%, 182/870). Other common factors included background interference (7.6%, 66/870) and failed hybridization (6.1%, 53/870). Rare causes included nucleus damage (1.8%, 16/870), nucleus loss (1.1%, 10/870) and signal split/overlap (0.9%, 8/870).</p><p><b>CONCLUSION</b>Multi-round FISH can improve the utility of single nucleus in PGD and the former three rounds have the highest efficiency. The hybridization effect of CEP is better than other probe. Poor signal quality is the common cause of unidentification results.</p>

Gene expression profiles of peri-implantation endometrium of high responders during controlled ovarian hyperstimulation / 上海第二医科大学学报

Objective To investigate the gene expression profiles of peri-implantation endometrium of high responders during controlled ovarian hyperstimulation. Methods High responders with cancelled embryo-transfer during controlled ovarian hyperstimulation (high responder group, n=4) and healthy fertile volunteers (control group, n=3) were performed endometrial biopsies during peri-implantation. Histologic changes of endometrium were observed by HE staining, genes of differential expression were screened with microarrays Affymetrix U133A 2.0 and identified by Real-time PCR. The biological process analysis was performed by online biological information analysis tool PANTHER. Results The ehdometrium was in mid-secretory phase in control group, while development delay was found in some glandular organs in endometrium of high responder group. Three hundred and sixty-four genes of differential expression were screened, among which 233 were up-regulated genes and 131 were down-regulated genes. OPN, PLA2G2, DPPIV, IGFBP5 and SSAT were identified as endometrial function-related genes, whose Real-time PCR findings were positively correlated to gene signal values detected by microarray(r=0.44, P<0.01). PANTHER analysis indicated that genes of differential expression participated in the biological processes of cytokine signal transduction and immunological regulation. Conclusion Ovarian high response affects the gene expression profiles of peri-implantation endometrium, which may be one of the causes of sub-optimal endometrial receptivity.

Effects of serum estradiol levels during controlled ovarian hyperstimulation on outcomes of IVF-ET / 上海第二医科大学学报

Objective To analyse the effects of serum estradiol levels during controlled ovarian hyperstimulation (COH) on the outcomes of in vitro fertilization and embryo transfer (IVF-ET). Methods The clinical data of 472 patients undergoing IVF-ET with GnRH analogues recombinant FSH long protocol were retrospectively analysed. The area under the curve (AUC) of estradiol (E2) level was calculated during COH, and patients were categorized into groups according to the percentile of AUC of E2(AUCE2) during COH. The general characteristics and parameters related to the outcomes of IVF-ET were compared among groups. Results The 10th percentile and 90th percentile of AUCE2 were 3 347.0 pmoL/L and 14 414.3 pmol/L, respectively. Four hundred and seventy-two patients were divided into lower reaction group (AUCE2 3 347.0 pmol/L, n=48), normal reaction group (14 414.3 pmol/L>AUCE2 > 3 347.0 pmol/L, n=376) and higher reaction group (AUCE2≥14 413.3 pmol/L, n=48). There was no significant difference in age, body mass index, baseline follicle stimulating hormone level, time of treatment with gonadotropin, endometrium thickness on day of transfer and embryos transferred(P>0.05). Compared with lower reaction group and normal reaction group, the number of oocytes per retrieval and number of embryos frozen were significantly larger(P<0.01) and the mild/severe ovarian hyperstimulation syndrome rate was significantly higher in higher reaction group(P<0.05). There was no significant difference in fertilization rate, cumulative embryo score, high-grade embryo rate, clinical pregnancy rate and implantation rate among groups (P> 0.05). Conclusion Sustained snpraphysiological serum E2 levels during the COH process do not adversely affect the quality of oocytes and embryos, clinical pregnancy rate and implantation rate to some extent in IVF-ET.

Impact of body mass index on outcome of in vitro fertilization-embryo transfer in patients with polycystic ovarian syndrome / 上海第二医科大学学报

Objective To compare the clinical outcomes of in vitro fertilization-embryo transfer(IVF-ET) in polycystic ovary syndrome(PCOS) patients with different body mass index(BMI).MethodsEighty-seven PCOS patients who received IVF-ET were classified into four groups according to BMI: group A,BMI0.05).ConclusionThe doses of gonadotropins are increased in PCOS patients with increasing BMI.The number of usable embryo is significant decreased in PCOS patients with BMI≥25 kg/m2.