Absence of paternal accessory sex gland secretions disturbs epigenetic reprogramming and expression of Igf2 and Dlk1 in golden hamster embryos

Accessory sex gland (ASG) secretion is known to exert an effect on sperm that is heritable in hamster embryos. We hypothesized that ASG secretion changes the sperm epigenome, which in turn is propagated in sired embryos. To test our hypothesis, we produced male hamsters that were devoid of either all ASG (TX) or only the ventral lobe of the prostate gland (VPX). A sham-operated control group (SH) was also established. These males were mated with normal females; uterine sperm, fertilized oocytes, and pre-implantation embryos were harvested from the females after mating. Epididymal sperm were collected at the end of experiments. Immunofluorescent staining was performed on these harvested specimens using antibodies against 5-methylcytosine, Dnmt1, Dnmt3a, Dnmt3b, protamine 1, protamine 2, and aectyl-H4K5. Expression of Igf2 and Dlk1 were analyzed by real-time RT PCR and in situ hybridization. We demonstrated that the DNA methylation pattern changed dynamically in SH, TX, and VPX fertilized oocytes. In VPX and TX embryos, DNA demethylation was slower and remethylation was delayed when compared with SH embryos. In addition, Dnmt3b expression was also abnormal. When sperm from VPX and TX males were exposed to whole ASG secretion in vivo, the resulting embryos all methylated normally. Immunofluorescent staining revealed that there was no difference in protamine packaging of uterine sperm from VPX and TX males. The staining also showed a lower level of acetyl-H4K5 expression in the male pronuclei of TX produced embryos. Furthermore, the VPX and TX embryos also expressed higher levels Igf2, and Dlk1. We concluded that interactions between ASG and sperm affected: (1) histone acetylation in male pronuclei; (2) DNA methylation in fertilized oocytes; and (3) Igf2 and Dlk1 expression embryos.     

Requirement for ADAMTS-1 in extracellular matrix remodeling during ovarian folliculogenesis and lymphangiogenesis

Murine ovarian folliculogenesis commences after birth involving oocyte growth, somatic cell differentiation and structural remodeling of follicle stromal boundaries. The extracellular metalloproteinase ADAMTS-1 has activity against proteoglycans and collagen and is produced by the granulosa cells of ovarian follicles. Mice with ADAMTS-1 gene disruption are subfertile due to an unknown mechanism resulting in severely reduced ovulation. Here we show that ADAMTS-1 is necessary for structural remodeling during ovarian follicle growth. A significant reduction in the number of healthy growing follicles and corresponding follicle dysmorphogenesis commencing at the stage of antrum formation was identified in ADAMTS-1-/- ovaries. Morphological analysis and immunostaining of basement membrane components identified stages of follicle dysgenesis from focal disruption in ECM integrity to complete loss of follicular structures. Cells expressing the thecal marker Cyp-17 were lost from dysgenic regions, while oocytes and dispersed cells expressing the granulosa cell marker anti-mullerian hormone persisted in ovarian stroma. Furthermore, we found that the ovarian lymphatic system develops coincidentally with follicular development in early postnatal life but is severely delayed in ADAMTS-1-/- ovaries. These novel roles for ADAMTS-1 in structural maintenance of follicular basement membranes and lymphangiogenesis provide new mechanistic understanding of folliculogenesis, fertility and disease.     

Two-dimensional informative array testing

Summary Array-based group-testing algorithms for case identification are widely used in infectious disease testing, drug discovery, and genetics. In this article, we generalize previous statistical work in array testing to account for heterogeneity among individuals being tested. We first derive closed-form expressions for the expected number of tests (efficiency) and misclassification probabilities (sensitivity, specificity, predictive values) for two-dimensional array testing in a heterogeneous population. We then propose two "informative" array construction techniques which exploit population heterogeneity in ways that can substantially improve testing efficiency when compared to classical approaches that regard the population as homogeneous. Furthermore, a useful byproduct of our methodology is that misclassification probabilities can be estimated on a per-individual basis. We illustrate our new procedures using chlamydia and gonorrhea testing data collected in Nebraska as part of the Infertility Prevention Project.     

A review on the stem cell therapy and an introduction to exosomes as a new tool in reproductive medicine

Stem cell therapy and exosome therapy are the two experimental methods that are now at the center of attention. Various types of stem cells, especially mesenchymal stem cells and spermatogonial stem cells have been widely administrated in reproductive medicine. However, due to the limitation of injecting living cells, using their paracrine secretions such as exosomes seems to be a better option. Exosomes show regenerative, pro-angiogenic, anti-apoptotic, anti-inflammatory, anti-hypoxic, and anti-fibrotic characteristics. They can induce cell proliferation, cell viability, migration, oogenesis, spermatogenesis, capacitation, acrosome reaction, and embryonic implantation. Exosomes have shown promising results in regenerative medicine such as liver fibrosis, stroke, cardiac ischemia, and skin injuries. Exosomes have been used to treat reproductive diseases such as erectile dysfunction and primary ovarian insufficiency. However, the study of exosomes in reproductive medicine is limited. In this article, we are going to review some of the researches on the use of stem cells and exosomes in reproductive medicine and suggest administration of a combination of exosomes for alleviating the symptoms of endometriosis and asthenozoospermia based on previous studies.     

二甲双胍联合枸橼酸氯米芬对多囊卵巢综合征不孕患者性激素和胰岛素水平的影响

目的:探讨二甲双胍、枸橼酸氯米芬联合治疗对多囊卵巢综合征(PCOS)不孕患者性激素和胰岛素水平的影响。方法:选取我院于2017年1月到2018年7月期间收治的101例PCOS不孕患者,采用数字表法将患者随机分为对照组(n=50)和研究组(n=51),对照组给予枸橼酸氯米芬治疗,研究组在对照组基础上联合二甲双胍治疗,采用门诊复查等方式随访6个月,记录两组患者排卵率及妊娠率,比较两组患者治疗前、治疗后的性激素、胰岛素以及血管活性因子水平,记录不良反应发生情况。结果:两组患者治疗后黄体生成素(LH)、睾酮(T)、胰岛素(INS)、LH/卵泡刺激素(FSH)均较治疗前降低,且研究组低于对照组(P0.05)。研究组排卵率、妊娠率均高于对照组(P0.05)。两组患者治疗后血管紧张素(AT-Ⅱ)、血管内皮生长因子(VEGF)均较治疗前降低,且研究组低于对照组(P0.05)。两组不良反应发生率对比无统计学差异(P0.05)。结论:二甲双胍联合枸橼酸氯米芬治疗PCOS不孕,安全有效,可有效调节患者胰岛素、性激素水平,提高排卵率、妊娠率,改善血管活性因子水平。     

Co-occurrences of polymorphic heterochromatin regions of chromosomes and effect on reproductive failure

Although the polymorphic heterochromatin regions of chromosomes (heteromorphisms) have been extensively studied for their phenotypic effects on humans, co-occurrences of chromosome 1, 9, 16 and Y heteromorphisms and of acrocentric variants have never been studied on humans with an objective scoring system. Here we compared the frequencies of individual heteromorphisms on a total of 602, 768 and 224 patients with the indications of infertility, recurrent miscarriage and in vitro fertilization (IVF) failure, respectively and on 272 controls. Then we examined whether there were significant co-occurrences between heteromorphisms within and between the groups. There were no statistically significant differences in the frequencies of heteromorphisms between the groups. Both statistically significant and non-significant correlations were observed within the non-acrocentric and certain acrocentric heteromorphisms in each group. When these co-occurrences were examined between the groups, a 2.2 fold increased risk of IVF failure in males in the presence of either chromosome 13 or chromosome 21 variants was observed (95 %CI:1.1–4.2). We conclude that the simultaneous manifestations of heteromorphisms have no effect on reproductive failure. There seems to be a correlation between the non-acrocentric heteromorphisms (1qh+, 9qh+, 16qh + and Yqh+/-), which might be the result of complex interactions of formation of these heterochromatin regions. The correlations observed between certain acrocentric chromosomes might be related to satellite association and nucleolus formation. The increased risk observed in males with IVF failure in the presence of either chromosome 13 or 21 variants should be interpreted cautiously due to the heterogeneity of the group.     

调经促孕丸联合奥利司他治疗肥胖型多囊卵巢综合征不孕的临床研究

摘要 目的：探讨调经促孕丸联合奥利司他治疗肥胖型多囊卵巢综合征（PCOS）不孕的临床疗效。方法：选取2017年6月到2019年8月期间我院收治的肥胖型PCOS不孕患者120例,按照随机数字表法将患者分为对照组（n=60）和研究组（n=60）。对照组予以奥利司他治疗,研究组在对照组的基础上联合调经促孕丸治疗,比较两组患者疗效、激素水平、排卵率、妊娠率、血糖及不良反应。结果：研究组治疗3个疗程后的临床总有效率为90.00%（54/60）,高于对照组的70.00%（42/60）（P<0.05）。两组治疗3个疗程后睾酮（T）、卵泡刺激素（FSH）、促黄体生成素（LH）均较治疗前下降,且研究组低于对照组（P<0.05）。研究组的排卵率为85%（51/60）、妊娠率为65%（39/60）均高于对照组的65%（39/60）、46.67%（28/60）（P<0.05）。两组餐后2 h血糖(2hPG)、空腹血糖（FBG）、糖化血红蛋白（HbAlc）均较治疗前下降,且研究组低于对照组（P<0.05）。两组治疗期间不良反应发生率对比未见统计学差异（P>0.05）。结论：调经促孕丸联合奥利司他治疗肥胖型PCOS不孕患者,疗效显著,可有效改善患者激素水平、血糖、排卵率、妊娠率,且不增加不良反应发生率。     

输卵管梗阻性不孕症的常用治疗方法及研究进展

不孕症是妇科常见疾病,其发病率呈逐年上升的趋势,在我国其发病率约为7％～10％。在女性不孕症中,输卵管梗阻是最常见的原因之一,其中又以近端阻塞多见。目前,宫腹腔镜诊断输卵管通畅度已经成为诊断输卵管梗阻胜不孕症的金标准。但是,目前治疗输卵管梗阻性不孕症的方法及效果尚未统一,常用的治疗方法不能够对患者做出全面客观的评估及准确的治疗。宫腹腔镜手术联合导丝治疗输卵管梗阻性不孕症的方法是将内镜和介入技术联合使用,较好地解决了各种治疗方法的不足,该方法凭借其可视、微创、并发症少、输卵管再通率高等优点应用于,临床。本文将对女性输卵管梗阻性不孕症的常用治疗方法及研究进展做一综述。     

金黄色葡萄球菌致小鼠慢性输卵管炎性不孕模型制作

目的通过金黄色葡萄球菌直接感染小鼠输卵管,建立炎症致不孕的动物模型。方法用1×109/mL的金黄色葡萄球菌接种小鼠,制作慢性输卵管炎症模型,观察输卵管病理炎性改变以及小鼠的受孕情况。结果造模术75 d后,模型组小鼠受孕率、输卵管通畅率显著低于对照组（P〈0.001）。模型组肉眼观察输卵管有不同程度积水积脓、僵硬,输卵管与周围组织均有不同程度的粘连;病理学观察输卵管管腔被异物肉芽组织完全阻塞,全层均见大量慢性炎细胞浸润。结论输卵管内接种浓度1×109/mL的金黄色葡萄球菌可以成功建立小鼠输卵管炎性不孕模型。