Selection of reference genes for studies of porcine endometrial gene expression on gestational day 12

Comparing gene expression patterns in the endometrium on gestational day 12 (GD12) between Erhualian (ER) and Landrace × Large White (LL) pigs is helpful to understand the biological mechanisms of fecundity. Selecting genes that have stable expression levels as the internal standards in a comparative study is essential for identifying real gene-specific variation by quantitative RT-PCR (qRT-PCR). Five genes expressed in sow endometria on GD12 were evaluated for their suitability as internal control for relative quantification by qRT-PCR. These genes were beta-actin (ACTB), beta-2-microglobulin (B2M), phosphoglycerate kinase 1 (PGK1), RNA polymerase II polypeptide G (RPG), and ribosomal protein S20 (RPS20), which represent different functional classes. Our results indicated that ACTB, B2M, and PGK1 were not suitable as internal standards for normalization because of their huge variability between the two breeds. RPS20 and RPG were most stable, and the former is recommended to serve as the internal standard when the use of multiple housekeeping genes is unpractical.     

八种广西特有报春苣苔属植物开花生物学特性研究

广西是报春苣苔属植物的世界分布中心,特有种较多。该研究针对线叶报春苣苔(Primulina linearifolia)、条叶报春苣苔(P. ophiopogoides)、刺齿报春苣苔(P. spinulosa)、大根报春苣苔(P. macrorhiza)、线萼报春苣苔(P. linearicalyx)、癞叶报春苣苔(P. leprosa)、桂中报春苣苔(P. guizhongensis)、百寿报春苣苔(P. baishouensis),八种广西特有种的开花物候期、开花动态、子房发育情况、花粉活性等开花生物学特性进行了研究。结果表明:(1)八种广西特有报春苣苔属植物在野外及人工环境下部分种在花期、单株开花量、单花开放持续时间具有显著性差异。人工栽培植株在单株开花量、单花开放持续时间要优于野生植株。(2)不同物种的单花花期长短不同,为3～14 d。雄蕊在花蕾阶段已经基本完成生长。雌雄蕊成熟期不一致,雄蕊先于雌蕊成熟。(3)花冠半开放至完全打开阶段的花粉活性最高,均在90%以上。(4)花冠完全开放阶段的柱头可授性最强。(5)自花授粉不能结实,异花授粉可正常结实,自交种子发芽率在80%以上。综上结果不仅为解决珍稀物种的繁殖障碍和保存种质资源提供了数据支持,而且对后续提高报春苣苔属的杂交育种成功率具有重要意义。     

Endometrial and cervical polyps in 22 baboons (Papio sp.), 5 cynomolgus macaques (Macaca fascicularis) and one marmoset (Callithrix jacchus)

Background  Endometrial and cervical polyps are masses of endometrium or cervical epithelium that bulge into the uterine or cervical lumen. The physiopathology and contributing factors of endometrial polyps development are still unknown.
Methods  Clinical and pathology records of 28 non-human primates with histologically confirmed endometrial and cervical polyps were reviewed. Twenty-one baboons with endometrial polyps were evaluated for age at diagnosis, body weight, menstrual cycle length, presence of endometriosis and adenomyosis and number of offspring, cesarean sections, and stillbirths.
Results  Endometrial polyps in baboons were associated with increased age, decreased menstrual cycle lengths, endometriosis, and decreased parity. No differences were found for weight, adenomyosis, or number of cesarean sections or stillbirths.
Conclusions  Baboons are a promising model for the study of endometrial polyps because of their similarity to humans in both the development of endometrial polyps and association of many of the same risk factors.     

白杨派树种雌蕊柱头可授性及其检测方法的研究

以银腺杨、毛新杨和银毛杨为材料,采用定期授粉观察以及联苯胺-过氧化氢染色等方法,对白杨派树种雌蕊柱头可授性进行了研究,结果表明:在温室水培条件下,白杨树种雌蕊柱头的可授期一般在1~3 d内,其中毛新杨雌蕊柱头可授期比银腺杨和银毛杨相对短些.但3个树种进入最佳授粉时期时的柱头开裂角度约为180°,此时柱头发亮且表面有大量分泌物存在;在适宜授粉时期内,用联苯胺-过氧化氢染色后柱头变蓝,并有气泡产生。在育种实践中,可根据柱头形态以及联苯胺-过氧化氢染色后柱头颜色变化和气泡产生情况及时判别有关白杨派树种最佳授粉时期。     

降钙素对胚胎着床的影响机理

降钙素(calcitonin,CT)是甲状腺滤泡旁细胞分泌的一种含有32个氨基酸残基的肽类激素,是动物体内重要的调节钙磷代谢的内分泌因子。近年来的研究发现CT在胚胎着床过程中起着重要的作用。胚胎着床涉及到母体子宫和胚胎之间的复杂而精确的调控。在孕激素作用下,围着床期子宫内膜表达CT,CT与其膜受体结合后可激活腺苷酸环化酶(adenylate cyclase,AC)和磷脂酶(Cphospholipase C,PLC)等激酶的活性,促进细胞外Ca2 内流,从而促使子宫内膜和胚胎发生一系列的变化,有利于胚胎的植入。     

The effect of fludrocortisone on the uterine receptivity partially mediated by ERK1/2-mTOR pathway

Implantation of embryos needs endometrial receptivity. Mineralocorticoids is one of the causes influencing the implantation window. This study targeted to evaluation fludrocortisone different properties on endometrial receptivity. The objective of this study was to assess whether treatment with fludrocortisone could impact the expression of diverse genes and proteins that are involved in uterine receptivity in mice. In this study, 40 female adult BALB/c mice were used. The samples were allocated to four groups of ten. Control group (C) received: vehicle; fludrocortisone group (FCA): received 1.5 mg/kg fludrocortisone; PP242 group (PP242): received 30 mg/kg PP242; fludrocortisone+PP242 group (FCA+PP242): received fludrocortisone and PP242. Mice were killed on window implantation day after mating and confirmed pregnancy. The endometrial epithelium of mouse was collected to assess mRNA expression of leukemia inhibitory factor (LIF), mucin-1 (MUC1), heparin-binding epidermal growth factor (HB-EGF), (Msx.1), miRNA Let-7a, and miRNA 223-3p as well as protein expression of extracellular signal-regulated kinase 1/2 (ERK1/2), mammalian target of rapamycin (mTOR), and eukaryotic translation initiation factor 4E-binding protein 1 (4E-BP1) in the uterine using real-time PCR and western blot, respectively. In comparison with the control group, fludrocortisone administration upregulated the expression of LIF, HB-EGF, Msx.1, miRNA Let-7a, ERK1/2, and mTOR in the epithelial endometrium. The PP242-treated group demonstrated a significant rise in the expression of MUC1, miRNA 223-3p and a remarkable decline in ERK1/2 and p-4E-BP1 levels in comparison with the control group. Combination therapy of (FCA+PP242) resulted in a remarkable rise in LIF, Msx-1, HB-EGF, ERK1/2, and mTOR levels, in comparison with the PP242 group. Furthermore, combination therapy of (FCA+PP242) downregulated the expression of MUC1 in comparison with the PP242-treated group. According to the results, fludrocortisone affected uterine receptivity possibly by means of modulating the expression of genes involved in the uterine receptivity and activation of the ERK1/2-mTOR pathway.     

Aberrant endometrial steroid receptor expression in in-vitro maturation cycles despite hormonal luteal support: A pilot study

Clinical outcomes of fresh embryo transfer in non-hCG triggered in vitro maturation (IVM) cycles are inferior compared to vitrified-warmed embryo transfer. This is a prospective observational pilot study in a consecutive cohort of 31 polycystic ovary syndrome (PCOS) patients and 37 normo-ovulatory egg donors who underwent IVM without fresh embryo transfer between July 2009 and June 2014. All subjects received 150 IU of highly purified menotropin (HP-hMG) daily for three days. On cycle day 6, all patients started transdermal oestradiol (E2) at a daily dose of 9 mg. There was no human chorionic gonadotropin (hCG) trigger before oocyte retrieval (OR). Vaginal micronized progesterone was commenced on the evening after OR, at a daily dose of 600 mg. Additional luteal phase support (LPS) was administered as follows: Group A: no additional LPS; Group B: 1500 IU of hCG administered 4 h after OR and Group C: 5000 IU of hCG administered 4 h after OR + an additional injection of 5000 IU of hCG 1 day before endometrial biopsy. Endometrial biopsy for histology and immunohistochemistry (IHC) was performed on day 5 or 6 after OR. Instead of being downregulated, both PR-B and ERα in endometrial glands and stroma were moderately to strongly expressed in all three protocols, suggesting that the mid-luteal histological signature of endometrial receptivity is deficient in a non-hCG-triggered IVM cycle. Poor clinical outcomes after fresh embryo transfer following IVM are probably related to inappropriate endometrial development which may be linked to the short follicular phase of IVM cycles.     

Development of a high-throughput assay for human proprotein convertase 5/6 for detecting uterine receptivity

Embryo implantation requires a healthy embryo and a receptive uterus. In women, the inner lining of the uterus, the endometrium, remains in a hostile state and becomes receptive for embryo implantation for only a short period during each menstrual cycle. Determining endometrial receptivity is vital in in vitro fertilization (IVF) treatment because the timing of embryo transfer needs to be synchronized with endometrial receptivity. We have previously demonstrated that proprotein convertase 5/6A (PC6) is highly expressed in the receptive endometrium and that PC6 is critical for receptivity establishment in women. Furthermore, endometrial PC6 is secreted into the uterine fluid, and levels correlate with receptivity status. Detection of PC6 in uterine fluids, therefore, would provide a nonsurgical assessment of endometrial receptivity. However, to date no assays are available for human PC6. In this study, we produced three PC6 monoclonal antibodies (mAbs) and developed a sandwich enzyme-linked immunosorbent assay (ELISA) for PC6 detection in human uterine fluids. The PC6 mAbs were confirmed to be highly specific to PC6, and the ELISA detected PC6 in human uterine fluids with a significantly higher level during the receptive phase. This newly established PC6 ELISA provides an important tool in the development of noninvasive strategies to detect endometrial receptivity in women.