Effect of SARS-CoV-2 infection on semen parameters in sperm bank volunteers with normal sperm concentration.

In this study, we aimed to assess the effect of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection on semen parameters. The study comprised 110 sperm volunteers who self-reported SARS-CoV-2 infection from the Human Sperm Bank of the Center for Reproductive Medicine, Shandong University (Jinan, China). The volunteers had normal sperm concentration before infection. Each volunteer provided semen samples before and after infection. We selected 90 days after infection as the cutoff point. Semen parameters within 90 days after infection of 109 volunteers (group A) were compared with semen parameters before infection. Moreover, semen parameters on or after 90 days after infection of 36 volunteers (group B) were compared with semen parameters before infection. Furthermore, based on whether the volunteers had completed the three-dose SARS-CoV-2 vaccination booster, volunteers in group A and B were further divided into two subgroups separately. Semen parameters were compared before and after infection in each subgroup. Our results showed that in this cohort population, the semen quality in volunteers with normal sperm concentrations before infection decreased after SARS-CoV-2 infection within 90 days, while the semen quality returned to preinfection levels after 90 days. The completion of a three-dose SARS-CoV-2 vaccination booster may exert a protective effect on semen quality after infection.

Extracellular vesicles from iPSC-MSCs alleviate chemotherapy-induced mouse ovarian damage via the ILK-PI3K/AKT pathway.

Chemotherapy can significantly reduce follicle counts in ovarian tissues and damage ovarian stroma, causing endocrine disorder, reproductive dysfunction, and primary ovarian insufficiency (POI). Recent studies have suggested that extracellular vesicles (EVs) secreted from mesenchymal stem cells (MSCs) exert therapeutic effects in various degenerative diseases. In this study, transplantation of EVs from human induced pluripotent stem cell-derived MSCs (iPSC-MSC-EVs) resulted in significant restoration of ovarian follicle numbers, improved granulosa cell proliferation, and inhibition of apoptosis in chemotherapy-damaged granulosa cells, cultured ovaries, and in vivo ovaries in mice. Mechanistically, treatment with iPSC-MSC-EVs resulted in up-regulation of the integrin-linked kinase (ILK) -PI3K/AKT pathway, which is suppressed during chemotherapy, most likely through the transfer of regulatory microRNAs (miRNAs) targeting ILK pathway genes. This work provides a framework for the development of advanced therapeutics to ameliorate ovarian damage and POI in female chemotherapy patients.

Growth hormone supplementation ameliorates blastocyst euploidy rates and improves pregnancy outcomes in women undergoing preimplantation genetic testing for aneuploidy cycles.

Background: Growth hormone (GH) supplementation has been shown to improve oocyte quality and live birth, but few studies have examined whether GH can reduce embryonic aneuploidy. Chromosomal abnormalities in preimplantation embryos have been regarded as the principal cause of implantation failure and miscarriage, and an increased percentage of aneuploid embryos has been observed in patient cohorts with unexplained recurrent pregnancy loss (RPL), recurrent implantation failure (RIF), and advanced maternal age. Methods: This prospective cohort study was conducted on women whose previous PGT-A cycle ended up with no transferrable blastocysts, or the aneuploidy rate was above 50% and no live birth was acquired. The participants were divided into GH co-treatment and comparison groups according to whether GH was administered in the subsequent PGT-A cycle. In addition, within the GH co-treatment group, the previous failed cycle constituted the self-control group. Results: 208 women were recruited in the study (GH co-treatment group: 96 women, comparison group: 112 women). Compared to the self-control and comparison groups, the rate of euploid blastocysts was significantly higher in the GH co-treatment group (GH vs self-control: 32.00% vs 9.14%, odds ratio [OR]: 4.765, 95% confidence interval [CI]: 2.420-9.385, P < 0.01; GH vs comparison: 32.00% vs. 21.05%, OR: 1.930, 95% CI: 1.106-3.366, P = 0.021), and their frozen embryo transfers resulted in more pregnancies and live births. In the subgroup analysis, for the <35 and 35-40 years groups, the euploidy rate in the GH co-treatment group was significantly higher than those in the self-control and comparison groups, but in the >40 years group, there was no difference in euploidy rate. Conclusion: Our study presents preliminary evidence that GH supplementation may ameliorate blastocyst aneuploidy and improve pregnancy outcomes in women who have previously experienced pregnancy failures along with high aneuploidy rates, particularly in those younger than 40 years. Therefore, the use of GH in such women should be considered. However, considering the limited sample size and mixed indications for PGT-A, further scientific research on the underlying mechanism as well as clinical trials with larger sample sizes are needed to confirm the effects and optimal protocols.

[Clinical and Cytogenetical Characteristics in Acute Myeloid Leukemia with Myelodysplasia-Related Changes].

OBJECTIVE: To explore the clinical and cytogenetic characteristics of acute myeloid leukemia with myelodysplasia-related changes (AML-MRC) based on morphology define. METHODS: A total of 180 newly diagnosed acute myeloid leukemia (AML) patients were enrolled and retrospectively analyzed, and marrow cell morphology of 126 patients were re-evaluated. The clinical and cytogenetic characteristics, including ages, sex, WBC count, HGB level, PLT count, blasts percentage, abnormal karyotype detection rate of the patients in AML with multilineage dysplasia (AML-MRC-1), secondary AML from myelodysplastic/ myeloproliferative neoplasms (MDS/MPN) (AML-MRC-2), and AML not otherwise specified (AML-NOS) groups were investigated. RESULTS: There was no significant differences between the patients in three groups in terms of sex, age and platelet count (P=0.898, P=0.365, P=0.853), but AML-MRC-2 group (73.2%) was higher than AML-MRC-1 (60.0%) and AML-NOS (56.4%) in the percentages of patients over 60 years old (P=0.228); there were statistically significant differences on WBC count, HGB level, and blasts percentage (P=0.000, P=0.022, P=0.000, AML-MRC-2

Single-Oocyte Gene Expression Suggests That Curcumin Can Protect the Ovarian Reserve by Regulating the PTEN-AKT-FOXO3a Pathway.

A better understanding of the mechanism of primordial follicle activation will help us better understand the causes of premature ovarian insufficiency (POI), and will help us identify new drugs that can be applied to the clinical treatment of infertility. In this study, single oocytes were isolated from primordial and primary follicles, and were used for gene profiling with TaqMan array cards. Bioinformatics analysis was performed on the gene expression data, and Ingenuity Pathway Analysis was used to analyze and predict drugs that affect follicle activation. An ovarian in vitro culture system was used to verify the function of the drug candidates, and we found that curcumin maintains the ovarian reserve. Long-term treatment with 100 mg/kg curcumin improved the ovarian reserve indicators of AMH, FSH, and estradiol in aging mice. Mechanistic studies show that curcumin can affect the translocation of FOXO3, thereby inhibiting the PTEN-AKT-FOXO3a pathway and protecting primordial follicles from overactivation. These results suggest that curcumin is a potential drug for the treatment of POI patients and for fertility preservation.

Endometrial cavity fluid is associated with deleterious pregnancy outcomes in patients undergoing in vitro fertilization/intracytoplasmic sperm injection: a retrospective cohort study.

BACKGROUND: The effects of endometrial cavity fluid (ECF) on in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) pregnancy outcomes following embryo transfer (ET) are still controversial. We conducted the present study to investigate whether the presence of ECF in infertile patients scheduled to undergo IVF or ICSI was associated with pregnancy outcomes. METHODS: A retrospective cohort study design was used. Among infertile patients undergoing IVF/ICSI, those with and without ECF were matched 1:1 using propensity score matching (PSM). After ensuring that the baseline levels of the two matched groups were consistent, the pregnancy and obstetrical outcomes of the two groups were compared. RESULTS: Patients with ECF had significantly lower clinical rates of pregnancy (1,061/1,862, 57% vs. 1,182/1,862, 63.5%; P<0.001), live birth (902/1,862, 48.4% vs. 1,033/1,862, 55.5%; P<0.001), biochemical pregnancy (1,182/1,862, 63.5% vs. 1,288/1,862, 69.2%; P<0.001), and embryo implantation (1,500/3,740, 40.1% vs. 1,661/3,740, 44.4%, P<0.001) than patients without ECF. Also, patients with ECF had a higher incidence of gestational diabetes (17/78, 22% vs. 8/94, 9%, P=0.014). However, there were no differences in gestational weeks at delivery or birth weight between the two groups. CONCLUSIONS: ECF was significantly associated with adverse pregnancy outcomes but showed no significant association with adverse obstetric outcomes (except for gestational diabetes).

Mutational analysis of the GATA4 gene in Chinese men with nonobstructive azoospermia.

As a crucial transcription factor for spermatogenesis, GATA-binding protein 4 (GATA4) plays important roles in the functioning of Sertoli and Leydig cells. Conditional knockout of GATA4 in mice results in age-dependent testicular atrophy and loss of fertility. However, whether GATA4 is associated with human azoospermia has not been reported. Herein, we analyzed the GATA4 gene by direct sequencing of samples obtained from 184 Chinese men with idiopathic nonobstructive azoospermia (NOA). We identified a missense mutation (c.191G>A, p.G64E), nine single-nucleotide polymorphisms (SNPs), and one rare variant (c.*84C>T) in the 3´ untranslated region (UTR). Functional studies demonstrated that the p.G64E mutation did not affect transactivation ability of GATA4 for spermatogenesis-related genes (claudin-11 and steroidogenic acute regulatory protein, Star), and the 3´ UTR rare variant c.*84C>T did not generate microRNA-binding sites to repress GATA4 expression. To our knowledge, this is the first report to investigate the association between GATA4 and azoospermia; our results indicate that mutations in GATA4 may not be pathogenic for NOA in Chinese men.

Corrigendum: Hormone-Like Effects of 4-Vinylcyclohexene Diepoxide on Follicular Development.

[This corrects the article DOI: 10.3389/fcell.2020.00587.].

Systemic changes in a mouse model of VCD-induced premature ovarian failure.

AIMS: Premature ovarian failure (POF) is a phenomenon in which the ovaries fail before the age of 40 years. Prior research has used a wide range of mouse models designed to reflect different causes of POF, including genetic factors, iatrogenic factors, and immune factors. The current study employed a mouse model of POF induced by 4-vinylcyclohexene diepoxide (VCD). VCD can specifically kill primordial and primary ovarian follicles, which destroys the follicular reserve and causes POF. The current study sought to specify and extend the applications of this model by examining the effect of timing and VCD dose and by exploring the effect of the model on systems outside of the ovaries. MATERIALS AND METHODS: A VCD-induced mouse model of POF was constructed using established methods (VCD injected continuously at a concentration of 160 mg/kg for 15 days). Evidence for a graded effect of VCD was observed using a range of concentrations, and the best windows for examining VCD's effects on follicles and associated tissues were identified. KEY FINDINGS: The mouse model used here successfully simulated two common complications of POF - emotional changes and decreased bone density. The model's application was then extended to examine the links between disease and intestinal microorganisms, and evidence was found linking POF to the reproductively relevant composition of the gut microbiota. SIGNIFICANCE: These findings provide novel methodological guidance for future research, and they significantly extend the applications and scope of VCD-induced POF mouse models.

Hormone-Like Effects of 4-Vinylcyclohexene Diepoxide on Follicular Development.

BACKGROUND: 4-vinylcyclohexene diepoxide (VCD) has long been considered a hazardous occupational chemical that promotes ovarian failure. However, VCD is also used as a research compound to chemically induce animal models of premature ovarian insufficiency (POI), and in related work we unexpectedly found that VCD apparently exhibits both dose- and duration-dependent opposing, hormone-like effects on the maintenance of the primordial follicle pool, follicle development, and ovulation induction. RESULTS: We conducted experiments with cultured murine ovaries and performed transplantation experiments using postnatal day (PD) 2 and PD12 mice and found that low-dose, short-term exposure to VCD (VCDlow) actually protects the primordial/primary follicle pool and improves the functional ovarian reserve (FOR) by disrupting follicular atresia. VCDlow inhibits follicular apoptosis and regulates the Pten-PI3K-Foxo3a pathway. Short-term VCD exposure in vivo (80 mg/kg, 5 days) significantly increases the number of superovulated metaphase II oocytes, preovulatory follicles, and corpus luteum in middle-aged mice with diminished ovarian reserve (DOR). We demonstrate that low-dose but not high-dose VCD promotes aromatase levels in granulosa cells (GCs), thereby enhancing the levels of estradiol secretion. CONCLUSION: Our study illustrates a previously unappreciated, hormone-like action for the occupational "ovotoxin" molecule VCD and strongly suggests that VCDlow should be explored for its potential utility for treating human ovarian follicular development disorders, including subfertility in perimenopausal women.

Pten-mediated Gsk3ß modulates the naïve pluripotency maintenance in embryonic stem cells.

Mouse embryonic stem cells (ESCs) are isolated from the inner cell mass of blastocysts, and they exist in different states of pluripotency-naïve and primed states. Pten is a well-known tumor suppressor. Here, we generated Pten-/- mouse ESCs with the CRISPR-Cas9 system and verified that Pten-/- ESCs maintained naïve pluripotency by blocking Gsk3ß activity. Serum/LIF and 2i (MAPK and GSK3 inhibitors) conditions are commonly used for ESC maintenance. We show that the Pten-inhibitor SF1670 contributed to sustaining mouse ESCs and that Pten activation by the S380A, T382A, and T383A mutations (Pten-A3) suppressed the pluripotency of ESCs. The in vivo teratoma formation ability of SF1670-treated ESCs increased, while the Pten-A3 mutations suppressed teratoma formation. Furthermore, the embryoid bodies derived from Pten-deficient ESCs or SF1670-treated wild-type ESCs showed greater expression of ectoderm and pluripotency markers. These results suggest that Pten-mediated Gsk3ß modulates the naïve pluripotency of ESCs and that Pten ablation regulates the lineage-specific differentiation.

The histone modification reader ZCWPW1 is required for meiosis prophase I in male but not in female mice.

Meiosis is a specialized type of cell division that creates haploid germ cells and ensures their genetic diversity through homologous recombination. We show that the H3K4me3 reader ZCWPW1 is specifically required for meiosis prophase I progression in male but not in female germ cells in mice. Loss of Zcwpw1 in male mice caused a complete failure of synapsis, resulting in meiotic arrest at the zygotene to pachytene stage, accompanied by incomplete DNA double-strand break repair and lack of crossover formation, leading to male infertility. In oocytes, deletion of Zcwpw1 only somewhat slowed down meiosis prophase I progression; Zcwpw1-/- oocytes were able to complete meiosis, and Zcwpw1-/- female mice had normal fertility until mid-adulthood. We conclude that the H3K4me3 reader ZCWPW1 is indispensable for meiosis synapsis in males but is dispensable for females. Our results suggest that ZCWPW1 may represent a previously unknown, sex-dependent epigenetic regulator of germ cell meiosis in mammals.

Novel Method of 3-Dimensional Graphical Representation for Proteins and Its Application.

In this article, we propose a 3-dimensional graphical representation of protein sequences based on 10 physicochemical properties of 20 amino acids and the BLOSUM62 matrix. It contains evolutionary information and provides intuitive visualization. To further analyze the similarity of proteins, we extract a specific vector from the graphical representation curve. The vector is used to calculate the similarity distance between 2 protein sequences. To prove the effectiveness of our approach, we apply it to 3 real data sets. The results are consistent with the known evolution fact and show that our method is effective in phylogenetic analysis.

Higher Prevalence of Endometrial Polyps in Infertile Patients with Endometriosis.

AIM: To study whether infertile patients with endometriosis have a higher prevalence of endometrial polyps, and to clarify the characteristics of the pathology of combined polyps. METHODS: Infertile patients who had undergone both hysteroscopy and laparoscopy in Reproductive Hospital Affiliated with Shandong University from January 2014 to May 2017 were enrolled. Patients with and without endometriosis, diagnosed by laparoscopy, were staged and included in the study group and control group, respectively, and the prevalence of polyps was compared. The pathological types of endometrial polyps were analyzed. RESULTS: A total of 414 cases were enrolled in the study group and 3,048 cases in the control group; polyps were diagnosed, with endoscopy, in 1,107 patients. Endometrial polyps were detected by hysteroscopy in 47.83% of the endometriosis group and 29.82% of the control group. The prevalence of endometrial polyps was significantly higher in the endometriosis group than in the control group (p < 0.001) but not significantly different between stages of endometriosis (p = 0.580). The pathological diagnosis included 899 endometrial polyps and 208 polypoid hyperplasia; 66.5% of endometrial polyps were combined with simple hyperplasia. CONCLUSIONS: The infertile patients with endometriosis had a higher prevalence of endometrial polyps, and those polyps are often combined with simple hyperplasia.

Identification of reference genes for qRT-PCR in granulosa cells of healthy women and polycystic ovarian syndrome patients.

Comparative gene expression analysis by qRT-PCR is commonly used to detect differentially expressed genes in studies of PCOS pathology. Impaired GC function is strongly associated with PCOS pathogenesis, and a growing body of studies has been dedicated to identifying differentially expressed genes in GCs in PCOS patients and healthy women by qRT-PCR. It is necessary to validate the expression stability of the selected reference genes across the tested samples for target gene expression normalization. We examined the variability and stability of expression of the 15 commonly used reference genes in GCs from 44 PCOS patients and 45 healthy women using the GeNorm, BestKeeper, and NormFinder statistical algorithms. We combined the rankings of the three programs to produce a final ranking based on the geometric means of their stability scores. We found that HPRT1, RPLP0, and HMBS out of 15 examined commonly used reference genes are stably expressed in GCs in both controls and PCOS patients and can be used for normalization in gene expression profiling by qRT-PCR. Future gene-expression studies should consider using these reference genes in GCs in PCOS patients for more accurate quantitation of target gene expression and data interpretation.

[Estimation of net primary productivity in arid region based on coupling model.] / 基于耦合模型的干旱区植被净初级生产力估算.

Net primary productivity (NPP), as the base for the research of matter recycling and energy flow in terrestrial ecosystem, is sensitive to the changes of environment and climate in arid region, and also is an important indicator of eco-environmental characteristics. Based on remote sensing (RS) and geographic information system (GIS), using meteorological data, eddy cova-riance data, Landsat 8 and MODIS data, this study coupled SEBAL model and light utility efficiency model to estimate the NPP of vegetation in Manas River Watershed, and the spatial pattern of NPP and the relationships between NPP and terrain factors (elevation and slope) were analyzed. Results showed that the estimated result of NPP in Manas River Watershed by coupling model was reasonable and could actually reflect the NPP of vegetation. The total annual NPP of vegetation and the mean annual NPP in Manas River Watershed in 2013 were 7066.72 Tg C·a-1 and 278.06 g C·m-2·a-1 respectively. With the variation of geomorphic type and land cover, the NPP changed remarkably from south to north in a trend of increase-decrease-increase-decrease pattern. The temporal variations of NPP were also obvious, with the NPP in July and August accounting for 52.2% of total annual NPP. With the increase of the elevation and slope, the mean annual NPP decreased as a whole with fluctuations induced by different land covers and environmental factors.

Predictors of Gestational Diabetes Mellitus in Chinese Women with Polycystic Ovary Syndrome: A Cross-Sectional Study.

AIMS: This study aims to explore the independent predictors of gestational diabetes mellitus (GDM) in Chinese women with polycystic ovary syndrome (PCOS). METHODS: This cross-sectional study analyzed primigravid women with PCOS and classified them as those with and without GDM. Independent risk factors and model performance were analyzed using multivariate logistic regression and the area under the curve (AUC) of receiver operating characteristic (ROC), respectively. RESULTS: Maternal body mass index, waist circumference, waist-to-hip ratio (WHR), fasting glucose, insulin, sex hormone-binding globulin (SHBG), homeostasis model assessment-insulin resistance (HOMA-IR) before pregnancy, gestation weight gain before 24 weeks and the incidence of family history of diabetes were different in the 2 groups. Logistic regression analysis showed that pre-pregnancy WHR, SHBG, HOMA-IR and gestation weight gain before 24 weeks were the independent predictors of GDM. ROC curve analysis confirmed that gestation weight gain before 24 weeks (AUC 0.767, 95% CI 0.688-0.841), pre-pregnant WHR (AUC 0.725, 95% CI 0.649-0.802), HOMA-IR (AUC 0.711, 95% CI 0.632-0.790) and SHBG levels (AUC 0.709, 95% CI 0.625-0.793) were the strong risk factors. CONCLUSIONS: In Chinese women with PCOS, factors of gestation weight gain before 24 weeks, pre-pregnant WHR, HOMA-IR and SHBG levels are strongly associated with subsequent development of GDM.

Coexistence of SFO-1 and NDM-1 ß-lactamase genes and fosfomycin resistance gene fosA3 in an Escherichia coli clinical isolate.

This study aims to characterize antimicrobial resistance and antimicrobial resistance genetic determinants of an Escherichia coli clinical isolate HD0149 from China in 2012. This strain displayed high-level resistance to cephalosporins, carbapenems, fluoroquinolones, aminoglycosides and fosfomycin. A range of antimicrobial resistance genes was detected responsible for its multiple antimicrobial resistances, involving the blaCMY-2, blaCTX-M-65, blaNDM-1, blaSFO-1, blaTEM-1, fosA3, rmtB, sul1 and sul2 genes. Four amino acid substitutions were detected in the quinolone resistance-determining regions (QRDRs) of GyrA (S83L and D87N), ParC (S80I) and ParE (S458A). Conjugation experiments revealed two multiresistance plasmids present in E. coli HD0149. The blaSFO-1 gene associated with blaNDM-1 gene was located in a 190 kb IncA/C plasmid and the blaCTX-M-65, fosA3 and rmtB genes were located in a 110 kb IncF plasmid. This is the first identification of the blaSFO-1 gene in an E. coli isolate and on a conjugative IncA/C plasmid. This may dramatically enhance the international prevalence and dissemination of blaSFO-1 among Enterobacteriaceae.

The association between testicular microlithiasis and semen parameters in Chinese adult men with fertility intention: experience of 226 cases.

OBJECTIVE: To investigate the association between testicular microlithiasis (TM) and semen parameters in Chinese adult men with fertility intention. MATERIALS AND METHODS: We retrospectively reviewed the ultrasonography results of the reproductive system of 16,204 consecutive adult male patients in our hospital with fertility intention from November 2012 to October 2013. TM was diagnosed by scrotal ultrasonography. Patients with TM were divided into classic testicular microlithiasis (CTM) or limited testicular microlithiasis (LTM). The clinical data of CTM, LTM, and non-TM groups, especially of patients in whom sperms were found in semen analysis, were collected and analyzed. RESULTS: There were 226 men (1.39%) diagnosed with TM. The mean age was 28.96 ± 5.12 years (range, 21-46 years), whereas mean testicular volume was 15.38 ± 4.90 mL (range, 1.62-31.23 mL). CTM and LTM were detected in 141 (62.39%) and 85 patients (37.61%), respectively. Among 200 patients who underwent semen analysis, sperms were found in 159 men (79.5%; 97 men with CTM and 62 men with LTM). One hundred and twenty cases without TM (ie, non-TM group) were collected in the control group. Sperm concentration, total motility, and percentage of progressively motile of CTM, LTM, and non-TM groups was (38.01 ± 31.58 million/mL vs 52.31 ± 33.26 million/mL vs 67.16 ± 36.94 million/mL; P <.001), (46.03 ± 23.69% vs 55.37 ± 24.16% vs 62.08 ± 20.45%; P <.001), and (35.88 ± 20.17% vs 43.15 ± 21.08% vs 47.10 ± 17.84%; P <.001), respectively. CONCLUSION: TM is associated with worse semen parameters in adult men with fertility intention. The extent of microlithiasis correlates inversely with semen parameters.