Prolonged HCG action affects angiogenic substances and improves follicular maturation, oocyte quality and fertilization competence in patients with polycystic ovarian syndrome

BACKGROUND: The aim of this study was to determine whether, in polycystic ovarian syndrome (PCOS) patients, HCG action prolonged for 4 h improves the action of angiogenic substances [ovarian renin angiotensin system and vascular endothelial growth factor (VEGF)], and consequently follicular maturation, oocyte quality and oocyte fertilization competence. METHODS: In this prospective study 20 patients with PCOS undergoing IVF were included. Oocyte retrieval was carried out either 34 or 38 h after HCG administration. Each follicle was analysed for prorenin, active renin, VEGF and estradiol. Oocytes were evaluated for quality (mature, immature, degenerated oocytes), as were the embryos (low or high). RESULTS: In the HCG +38 h group there were 245 follicles, and in the HCG +34 h group 240 follicles. In the HCG +38 h group, log active renin was lower (2.78 +/- 0.20 versus 2.91 +/- 0.25; P < 0.001) and VEGF higher (2276.0 +/- 790.1 versus 1946.6 +/- 954.5 pg/ml; P < 0.001). The odds ratio for obtaining oocytes from follicles was 1.6 [95% confidence interval (CI) 1.1-2.6; P = 0.02], and for developing high quality embryos 7.6 (95% CI 2.8-20.9; P < 0.001) in favour of the HCG +38 h group. CONCLUSIONS: Follicular maturation and oocyte quality are related to the intrafollicular influences of active renin and VEGF in a time-dependent manner after HCG administration, whereas fertilization competence is related to VEGF only.     

Mitochondrial DNA content and 4977 bp deletion in unfertilized oocytes

Previous studies analysing the incidences of mitochondrial DNA (mtDNA) deletions and mtDNA content in unfertilized oocytes in relation to donors' age have been controversial. The objective of the study was to compare these two parameters in unfertilized oocytes and relate them to the donors' age. Fifty-two women donated 155 unfertilized metaphase II (MII) oocytes. The incidence of 4977 bp deletion was 34.6%, and the mtDNA copy number was 598 350 +/- 265 862. Women >or=35 years of age had a significantly higher incidence of 4977 bp deletion, lower mtDNA copy number, higher FSH level and poorer ovarian response when compared with younger women. The mtDNA copy number was negatively correlated with the donor's age. The higher incidence of mtDNA deletion and lower mtDNA copy number in older women suggested that these two parameters may reflect ovarian ageing.     

An immunocytochemical study of the germinal layer vasculature in the developing fetal brain using Ulex europaeus 1 lectin

The characteristics of the germinal matrix vasculature were studied in the developing fetal brain using immunocytochemical methods. A preliminary comparative immunocytochemical study was made on six fetal brains to compare endothelial staining by Ulex europaeus I lectin with that of antibody to Factor VIII related antigen. Ulex was found to stain germinal layer vessels better than Factor VIII related antigen. Subsequently, the germinal layers of a further 15 fetal and preterm infant brains ranging from 13 to 35 weeks' gestation were stained with Ulex europaeus I to demonstrate the vasculature. With increasing gestation, there was a gradual increase in vessel density, particularly of capillaries. This was not a uniform process. A plexus of capillaries was prominent immediately beneath the ependyma while the more central parts of the germinal matrix contained fewer, but often larger diameter, vessels. The variation in vessel density which was a feature of the later gestation brains may have implications for local blood flow and may be a factor in haemorrhage at this site.     

敌枯双对雄性小鼠生殖细胞分裂比率及精原细胞染色体畸变的影响

本文研究丁敌枯双对BALB/C雄性小鼠生殖细胞分裂比率及精原细胞染色体畸变的影响。将实验小鼠随机分为三组,即实验组(敌枯双组),阳性对照组(环磷酰胺组)和阴性对照组(双蒸水组)。结果发现:敌枯双能明显诱发精原细胞多倍体率和裂隙率增加,抑制终变期/中期Ⅰ和中期Ⅱ细胞的减数分裂比率,促进精原细胞有丝分裂比率。实验结果还提示:在遗传毒理检测中亚急性实验是必要的,并对实验结果进行了初步讨论。     

合体细胞滋养层细胞外囊泡阻止母体恶性肿瘤侵袭及转移至胎儿相关机制研究

目的探讨合体细胞滋养层细胞外囊泡(STB-EV)阻止母体恶性肿瘤侵袭、转移至胎儿的相关机制。 方法选择2015年6月,于四川大学华西第二医院进行剖宫产术分娩的1例妊娠合并宫颈癌(CCP)患者被肿瘤侵犯的胎盘组织为研究对象。宫颈癌细胞系SiHa细胞和人滋养层细胞系HTR-8细胞,均由本院西部妇幼医学研究院-分子与转化医学实验室馈赠。其中,本例CCP患者被肿瘤侵犯的胎盘组织制作组织切片后,于光学显微镜(×400)和电子显微镜(×2 000)下,观察其组织形态学、STB-EV及合体细胞滋养层(STB)旁宫颈癌细胞自噬性死亡情况等;而SiHa细胞与HTR-8细胞培养后,用于Transwell迁移实验与细胞划痕实验。将SiHa与HTR-8细胞共培养,纳入研究组;单独培养的SiHa细胞,纳入对照组。对2组细胞进行Transwell迁移实验及细胞划痕实验,2组Transwell小室穿膜细胞数、细胞迁移率等比较,采用成组t检验。本研究遵循的程序符合2013年新修订的《世界医学协会赫尔辛基宣言》要求。 结果①对本例CCP患者被肿瘤侵犯的胎盘组织制作组织切片后,于光学显微镜(×400)下观察结果显示,与STB相邻宫颈癌细胞自噬性死亡、凋亡和固缩性坏死显著。电子显微镜(×2 000)下观察结果显示,宫颈癌细胞累及胎盘绒毛组织时,STB分泌的STB-EV水平显著高于正常胎盘组织。②2组细胞Transwell迁移实验结果显示,研究组穿膜细胞数为(597.6±87.7)个/视野,显著低于对照组的(1 358.4±203.0)个/视野,并且差异有统计学意义(t＝14.490、P<0.001)。③2组细胞划痕实验结果显示,划痕后48 h时,研究组细胞迁移率为(26.6±3.8)%,显著低于对照组的(45.9±3.7)%,并且差异有统计学意义(t＝3.122、P＝0.035)。 结论STB分泌的STB-EV可能参与母体肿瘤细胞广泛自噬性死亡,从而阻止母体肿瘤细胞侵袭胎盘、影响胎儿发育。     

Predictive model containing PI-RADS v2 score for postoperative seminal vesicle invasion among prostate cancer patients

BackgroundSeminal vesicle invasion (SVI) is considered to be one of most adverse prognostic findings in prostate cancer, affecting the biochemical progression-free survival and disease-specific survival. Multiparametric magnetic resonance imaging (mpMRI) has shown excellent specificity in diagnosis of SVI, but with poor sensitivity. The aim of this study is to create a model that includes the Prostate Imaging Reporting and Data System version 2 (PI-RADS v2) score to predict postoperative SVI in patients without SVI on preoperative mpMRI.MethodsA total of 262 prostate cancer patients without SVI on preoperative mpMRI who underwent radical prostatectomy (RP) at our institution from January 2012 to July 2019 were enrolled retrospectively. The prostate-specific antigen levels in all patients were <10 ng/mL. Univariate and multivariate logistic regression analyses were used to assess factors associated with SVI, including the PI-RADS v2 score. A regression coefficient-based model was built for predicting SVI. The receiver operating characteristic curve was used to assess the performance of the model.ResultsSVI was reported on the RP specimens in 30 patients (11.5%). The univariate and multivariate analyses revealed that biopsy Gleason grade group (GGG) and the PI-RADS v2 score were significant independent predictors of SVI (all P<0.05). The area under the curve of the model was 0.746 (P<0.001). The PI-RADS v2 score <4 and Gleason grade <8 yielded only a 1.8% incidence of SVI with a high negative predictive value of 98.2% (95% CI, 93.0–99.6%).ConclusionsThe PI-RADS v2 score <4 in prostate cancer patients with prostate-specific antigen level <10 ng/mL is associated with a very low risk of SVI. A model based on biopsy Gleason grade and PI-RADS v2 score may help to predict SVI and serve as a tool for the urologists to make surgical plans.     

Transfer of germinal vesicle to ooplasm of young mice could not rescue ageing-associated chromosome misalignment in meiosis of oocytes from aged mice

BACKGROUD: Transferring a germinal vesicle (GV) from an aged woman's oocyte into ooplasm from a younger woman has been proposed as a possible way to overcome the problem of age-related decline in female fertility. Here we assessed this possibility by determining whether ooplasts derived from young mice could rescue ageing-associated chromosome misalignment in meiosis of oocytes from aged mice. METHODS: Three groups of reconstructed oocytes, young GV-young cytoplast (group YY), aged GV-young cytoplast (group AY), and young GV-aged cytoplast (group YA), were created by micromanipulation and electrofusion. RESULTS: Nuclear transplantation was successful in 89.8-94.4% of GV-ooplast complexes, and maturation rate of the reconstructed oocytes was 93.5-97.9%. Confocal microscopy analysis showed a significantly higher rate (49.2%) of chromosome misalignment in ageing mice than in young mice (16.9%), and 57.1% of oocytes in group AY exhibited chromosome misalignment, while the abnormality rate in groups YY and YA was 16.3 and 16.7% respectively. Calcium imaging showed that the three groups of reconstructed oocytes exhibited a similar pattern of calcium oscillations upon stimulation with bovine sperm extracts. Fertilization rate and developmental capacity to 2-cell embryos were also similar among the three groups of oocytes. CONCLUSIONS: Our findings suggest that: (i) the ooplasm from young mice could not rescue ageing-associated chromosome misalignment in meiosis of GV from aged mice; and (ii) behaviour of chromosome alignment over metaphase spindle is predominantly determined by GV material.     

突触小泡膜表面突起的研究

本文首次报道了用透射电镜观察大白鼠的大脑皮质和海马内的突触小泡膜表面突起的超微结构，发现神经终末内的突触小泡膜表面有短突起的微细结构，借此使小泡与小泡、小泡与突触前膜、小泡与骨架发生联系，为突触小泡的转运、入坞和释放的分子神经生物学研究提供了超微形态学的证据     

Synthetic Evolution of a Supramolecular Harpooning Mechanism to Immobilize Vesicles at Antifouling Interfaces

The immobilization of vesicles has been conceptualized as a method to functionalize biointerfaces. However, the preservation of their integrity post immobilization remains a considerable challenge. Interfacial interactions can cause vesicle rupture upon close surface contact and non-specific protein adsorption impairing surface functions. To date, immobilization of vesicles has relied solely on either entrapment or prior modification of vesicles, both of which require laborious preparation and limit their applications. This work develops a bioinspired strategy to pin vesicles without prior modification while preserving their intact shape. This work introduces antifouling diblock copolymers and ultrathin surface-attached hydrogels containing a brush-like interface consisting of a bottle brush copolymer of N-(2-hydroxypropyl) methacrylamide (HPMA) and N-(3-methacrylamidopropyl)-N,N-dimethyldodecan-1-aminiumiodide (C12+). The presence of positive charges generates an attractive force that pulls vesicles toward the surface. At the surface, the amphiphilic properties of the combs facilitate their insertion into the membrane, mimicking the harpooning mechanism observed in antimicrobial peptides. Importantly, the antifouling poly(HPMA) backdrop serves to safeguard the vesicles by preventing deformation and breakage. Using a combination of thermodynamic analysis, surface plasmon resonance, and confocal laser scanning microscopy, this work demonstrates the efficiency of this biomimetic system to capture vesicles while maintaining an antifouling interface necessary for bioapplications.     

Epidural anesthesia in anin vitro fertilization and embryo transfer program

Objectives To investigate the potential advantages of epidural anesthesia in an in vitro fertilization and embryo transfer program.Study Design Between January 1991 and December 1992, 148 infertile patients underwent transvaginal ultrasound guided oocyte retrieval. A total of 44 patients (group A) had 50 retrievals under epidural anesthesia, and 104 patients (group B, control group) 112 retrievals under intravenous sedation (propofol) with mask-assisted ventilation (nitrous oxide).Results In group A satisfactory anesthesia was achieved in 49 of the 50 cases (98%); one patient required additional intravenous administration of propofol owing to extreme anxiety. No complications were observed. Adversely, in group B nausea was observed in 16 cases (14%) and nausea and vomiting in 7 cases (6%). In group A the fertilization, cleavage and pregnancy rates were 67.2%, 92% and 20% respectively, while in group B the rates were 69.3%, 93% and 19.6% respectively; the differences are not statistically significant.Conclusions Epidural anesthesia is an effective method for transvaginal oocyte retrieval but does not improve the treatment outcome as compared to an intravenous sedation with mask-assisted ventilation using propofol and nitrous oxide.