The Deletion of US3 Gene of Pseudorabies Virus (PRV) ΔgE/TK Strain Induces Increased Immunogenicity in Mice.

Re-emerging pseudorabies (PR) caused by pseudorabies virus (PRV) variant has been prevailing among immunized herds in China since 2011, indicating that commercially available PR vaccine strains couldn't provide complete protection against novel, epidemic PRV variant. Before this study, a gE/TK-gene-deleted virus (PRV ΔgE/TK) was constructed from PRV QYY2012 variant through homologous recombination and Cre/LoxP system. Here, PRV ΔgE/TK/US3 strain was generated by deleting US3 gene based on PRV ΔgE/TK strain using the same method. The growth characteristics of PRV ΔgE/TK/US3 were analogous to that of PRV ΔgE/TK. Moreover, the deletion of US3 gene could promote apoptosis, upregulate the level of swine leukocyte antigen class I molecule (SLA-I) in vitro, and relieve inflammatory response in inoculated BALB/c mice. Subsequently, the safety and immunogenicity of PRV ΔgE/TK/US3 was evaluated as a vaccine candidate in mice. The results revealed that PRV ΔgE/TK/US3 was safe for mice, and mice vaccinated with PRV ΔgE/TK/US3 could induce a higher level of PRV-specific neutralizing antibodies and cytokines, including IFN-γ, IL-2 and IL-4, also higher level of CD8+ CD69+ Tissue-Resident Memory T cells (TRM). The results show that the deletion of US3 gene of PRV ΔgE/TK strain could induce increased immunogenicity, indicating that the PRV ΔgE/TK/US3 strain is a promising vaccine candidate for preventing and controlling of the epidemic PR in China.

The Role of Abnormal Uterine Junction Zone in the Occurrence and Development of Adenomyosis.

Adenomyosis is a benign disease with a malignant behavior, bothering a lot of women at reproductive age who suffer from increased menstruation, prolonged menstruation, progressive dysmenorrhea, and infertility. At present, there is no effective treatment for adenomyosis. It seriously affects the life quality of these patients. However, the pathogenesis of adenomyosis is not yet clear. Recently, uterus junctional zone, defined as the inner 1/3 of myometrium between endometrium and myometrium, has gained broad attention. As is reported, the structure and function disorder of uterus junctional zone may play an important part in the occurrence and development of adenomyosis. In this issue, the present study generally reviews the role of uterine junction zone and the related mechanisms involved in adenomyosis, such as the local micro-damage, the formation of inflammatory and hypoxic microenvironment, changes of cytokines, and abnormalities of miRNA as well as signal pathways. It will provide new insights and potential therapeutic target strategies for clinical strategies in the management of adenomyosis.

Blastocyst culture of non-top-quality cleavage embryos may increase the risk of anembryonic pregnancy following in vitro fertilization: a retrospective cohort study.

Background: Anembryonic pregnancy (AP) is the most severe dysmorphogenesis of human embryo development and a frequent presentation of early pregnancy loss (EPL). Studies have analyzed the association between assisted reproductive technologies (ART) and EPL. However, the specific relationship between ART and AP has not been fully elucidated. Several studies suggested that non-genomic anomalies might be related to AP and ART might increase the risk of epigenetic changes, thus possibly detecting some associations between ART and AP. Our study aims to find out any possible risk factors of AP in ART treatments, and translate the results into clinical practice. Methods: A retrospective cohort study was conducted in Nanfang Hospital. Data from 1,765 singleton pregnancies following fresh or frozen-thawed embryo transfer from January 2014 to December 2017 were collated with the inclusion of EPLs and normal live births (NLB). Participants were divided into three groups: NLB (full-term birth with normal body weight infants), EPL (spontaneous pregnancy loss prior to 13 weeks gestation) with embryos (EE), and APs (embryonic pole was invisible in two consecutive ultrasound examinations). The basic characteristics of the patients and the association between ART-related variables and AP were analyzed using one-way analysis of variance (ANOVA) and multivariable logistic regression model, respectively. Products of conception (POC) from AP and EE patients received karyotype analysis using multiplex ligation-dependent probe amplification (MLPA). Results: Blastocyst culture of non-top-quality cleavage stage embryos almost doubled the percentage of AP in EPL (45.9% vs. 24.4%, P=0.037), and the normal euploid rate was significantly higher in the AP group (50.5% vs. 32.3%, P=0.003). Using multivariable logistic regression model, we found that blastocyst transfer and advanced maternal age might be risk factors for AP (OR >1, P<0.05). Deceased ß-HCG level might indicate its occurrence (OR <1, P<0.001) while CoQ10 supplementation might be a protective factor (OR <1, P<0.001). Conclusions: The occurrence of AP may be due to epigenetic abnormalities associated with advanced maternal age and extended in vitro embryo culture, while CoQ10 supplementation may be a potential method in preventing AP. Future multi-center prospective cohort studies should be conducted to verify these results.

Bisphenol A promotes autophagy in ovarian granulosa cells by inducing AMPK/mTOR/ULK1 signalling pathway.

BACKGROUND: Bisphenol A (BPA) is a widespread endocrine-disrupting chemical with estrogen like effects, which could interfere with the human reproductive system by disrupting the normal function of granulosa cells (GCs) leading to abnormal ovarian function. However, the mechanism of its toxicity on human GCs has not been clearly described thus far. METHODS: 106 normogonadotropic infertile women undergoing their first in-vitro fertilization-embryo transfer (IVF-ET) cycle were recruited. Urinary BPA level and the early outcomes of IVF-ET were analysed. Patients were divided to low and high BPA exposure groups using the median urinary BPA concentration as the cut-off value. In-vivo and in-vitro studies were conducted using mice and human granulosa cell line (KGN cells). Female Kunming mice approximately 6-8 weeks of age were poisoned with BPA at different dosages (1, 10 or 100 µg/kg) by oral gavage once daily for 2 weeks, while KGN cells were exposed to BPA at the concentration of 1, 10 or 100 nM for 24 h, 48 h or 72 h. BPA-induced ovarian morphologic changes were analysed by histopathology investigation. Cell viability and apoptosis were evaluated using CCK-8, TUNEL and flowcytometric, respectively. Hormone levels were determined using ELISA and the molecular mechanism studies were conducted using immunofluorescence, RT-PCR and western blots. RESULTS: The oocyte retrieval rate, maturation rate and embryo implantation rate significantly decreased with the higher level of urinary BPA concentration. Peak E2 level was lower in high BPA group, but no statistical significance could be observed. In BPA treated mice, cystic dilation of the follicles with a decreased number of GCs could be observed histopathologically. Decreased E2, P4 and AMH level and GCs autophagy could be detected both in-vivo and in-vitro with the activation of AMPK/mTOR/ULK1 signalling pathway. As being confirmed in KGN cells, phosphorylated AMPK and ULK1 increased while phosphorylated mTOR decreased, and by inhibition autophagy using knockdown of AMPK or 3-MA, adverse effects of BPA exposure in-vitro could be reversed. CONCLUSION: BPA exposure might abnormally influence human ovarian functions leading to abnormal folliculogenesis by activation of autophagy in GCs through AMPK/mTOR/ULK1 pathway.

Association of seminal angiotensinogen with sperm motility and morphology in male infertility.

Many researchers have shown that renin-angiotensin system (RAS) is involved in various important aspects of male reproduction. In this study, we assessed whether abnormal levels of seminal angiotensinogen (AGT) may be associated with semen parameters in infertile males. A total of 115 male patients were recruited, and semen parameters, seminal AGT and the electrolytes including K+ , Na+ , Cl- , P and Ca were evaluated. According to the World Health Organization (WHO) 2010 criteria, the patients were divided into two groups: G1 group with normal semen parameters (n = 42) and G2 group with subnormal semen parameters (n = 73). The level of seminal AGT was significantly higher in G2 group compared with G1 group. Moreover, the level of AGT was negatively correlated with the percentage of total motility (r = -.322, p = .000), progressive motility (PR) (r = -.339, p = .000) and morphologically normal forms (r = -.263, p = .004). This study suggests that elevated seminal AGT level is associated with increased risk of asthenospermia and teratozoospermia.