Abnormal Y chromosome detection in infertile males using multiplex ligation-dependent probe amplification.

Y chromosome abnormalities are the leading cause of male infertility. The clinical detection of abnormalities is necessary for appropriate genetic counselling. This study describes the prevalence, distribution and characteristics of Y chromosome abnormalities, which should be considered in the clinical management of infertile males. A total of 121 patients with oligozoospermia, 120 with azoospermia and 88 normal individuals were recruited between June 2019 and July 2021. Y chromosome microdeletions were assessed using multiplex ligation-dependent probe amplification (MLPA). The abnormal Y chromosome prevalence was 30.70%, and it was most common in patients aged 26-40 years. The frequencies of azoospermia factor (AZF) deletion, duplication and deletions/duplications were 19.76%, 9.42% and 1.52% respectively. The most common abnormalities were AZFc deletion (19.80%), AZFc partial deletion (40.59%) and AZFc partial duplication (17.82%). Oligozoospermia was associated with an increased incidence of AZF deletion. In the subgroup analysis, patients <30 years old with azoospermia exhibited elevated follicle-stimulating hormone levels and oestradiol. Moreover, the incidence of AZF deletion was higher in those with azoospermia (OR: 2.12; 95% CI: 1.05-5.28; p = 0.023) or oligozoospermia (OR: 2.54; 95% CI: 1.13-5.79; p = 0.008) than in normal individuals for ages ≥30 years.

Fetal Hyperthyroidism with Maternal Hypothyroidism: Two Cases of Intrauterine Therapy.

Fetal hyperthyroidism can occur secondary to maternal autoimmune hyperthyroidism. The thyroid-stimulating hormone receptor antibody (TRAb) transferred from the mother to the fetus stimulates the fetal thyroid and causes fetal thyrotoxicosis. Fetuses with this condition are difficult to detect, especially after maternal Graves disease therapy. Here, we present two cases of fetal hyperthyroidism with maternal hypothyroidism and review the assessment and intrauterine therapy for fetal hyperthyroidism. Both women were referred at 22+ and 23+ weeks of gestation with abnormal ultrasound findings, including fetal heart enlargement, pericardial effusion, and fetal tachycardia. Both women had a history of Graves disease while in a state of hypothyroidism with a high titer of TRAb. A sonographic examination showed a diffusely enlarged fetal thyroid with abundant blood flow. Invasive prenatal testing revealed no significant chromosomal aberration. Low fetal serum TSH and high TRAb levels were detected in the cord blood. Fetal hyperthyroidism was considered, and maternal oral methimazole (MMI) was administered as intrauterine therapy, with the slowing of fetal tachycardia, a reduction in fetal heart enlargement, and thyroid hyperemia. During therapy, maternal thyroid function was monitored, and the dosage of maternal levothyroxine was adjusted accordingly. Both women delivered spontaneously at 36+ weeks of gestation, and neonatal hyperthyroidism was confirmed in both newborns. After methimazole and propranolol drug treatment with levothyroxine for 8 and 12 months, both babies became euthyroid with normal growth and development.

Genome-wide association study of maternal plasma metabolites during pregnancy.

Metabolites are key indicators of health and therapeutic targets, but their genetic underpinnings during pregnancy-a critical period for human reproduction-are largely unexplored. Using genetic data from non-invasive prenatal testing, we performed a genome-wide association study on 84 metabolites, including 37 amino acids, 24 elements, 13 hormones, and 10 vitamins, involving 34,394 pregnant Chinese women, with sample sizes ranging from 6,394 to 13,392 for specific metabolites. We identified 53 metabolite-gene associations, 23 of which are novel. Significant differences in genetic effects between pregnant and non-pregnant women were observed for 16.7%-100% of these associations, indicating gene-environment interactions. Additionally, 50.94% of genetic associations exhibited pleiotropy among metabolites and between six metabolites and eight pregnancy phenotypes. Mendelian randomization revealed potential causal relationships between seven maternal metabolites and 15 human traits and diseases. These findings provide new insights into the genetic basis of maternal plasma metabolites during pregnancy.

Effects of HAR1 on cognitive function in mice and the regulatory network of HAR1 determined by RNA sequencing and applied bioinformatics analysis.

Background: HAR1 is a 118-bp segment that lies in a pair of novel non-coding RNA genes. It shows a dramatic accelerated change with an estimated 18 substitutions in the human lineage since the human-chimpanzee ancestor, compared with the expected 0.27 substitutions based on the slow rate of change in this region in other amniotes. Mutations of HAR1 lead to a different HAR1 secondary structure in humans compared to that in chimpanzees. Methods: We cloned HAR1 into the EF-1α promoter vector to generate transgenic mice. Morris water maze tests and step-down passive avoidance tests were conducted to observe the changes in memory and cognitive abilities of mice. RNA-seq analysis was performed to identify differentially expressed genes (DEGs) between the experimental and control groups. Systematic bioinformatics analysis was used to confirm the pathways and functions that the DEGs were involved in. Results: Memory and cognitive abilities of the transgenic mice were significantly improved. The results of Gene Ontology (GO) analysis showed that Neuron differentiation, Dentate gyrus development, Nervous system development, Cerebral cortex neuron differentiation, Cerebral cortex development, Cerebral cortex development and Neurogenesis are all significant GO terms related to brain development. The DEGs enriched in these terms included Lhx2, Emx2, Foxg1, Nr2e1 and Emx1. All these genes play an important role in regulating the functioning of Cajal-Retzius cells (CRs). The DEGs were also enriched in glutamatergic synapses, synapses, memory, and the positive regulation of long-term synaptic potentiation. In addition, "cellular response to calcium ions" exhibited the second highest rich factor in the GO analysis. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis of the DEGs showed that the neuroactive ligand-receptor interaction pathway was the most significantly enriched pathway, and DEGs also notably enriched in neuroactive ligand-receptor interaction, axon guidance, and cholinergic synapses. Conclusion: HAR1 overexpression led to improvements in memory and cognitive abilities of the transgenic mice. The possible mechanism for this was that the long non-coding RNA (lncRNA) HAR1A affected brain development by regulating the function of CRs. Moreover, HAR1A may be involved in ligand-receptor interaction, axon guidance, and synapse formation, all of which are important in brain development and evolution. Furthermore, cellular response to calcium may play an important role in those processes.

[Clinical study on 156 cases with hydrops fetalis].

OBJECTIVE: To investigate the ultrasound characteristics, etiology and prognosis in hydrops fetalis. METHODS: From September 2002 to May 2010, 156 hydrops fetalis presented in Shenzhen Maternity and Child Healthcare Hospital were studied retrospectively, including ultrasound characteristics, etiology, and prognosis. RESULTS: All of the 112 typical hydrops fetalis, 20 cases with isolated ascites, 8 cases with isolated pleural effusion, 7 cases with isolated pericardial effusion, 5 cases with isolated subcutaneous edema, 4 cases with isolated placental thickening were observed by ultrasonography. The major etiology and associated diagnosis consisted of 35.9% (56/156) of non-immune anemia, 9.6% (15/156) of cardiac abnormalities, 7.1% (11/156) of intrauterine infection, 6.4% (10/156) of twin problems, 5.8% (9/156) of meconium peritonitis, 5.1% (8/156) of thoracic-lung disease, 4.5% (7/156) of chromosomal abnormalities, 1.9% (3/156) of immune anemia. Alpha thalassemia was the most common non-immune anemia (96%, 54/56). An etiology and associated diagnosis could be determined in 81.4% (127/156) of cases. Follow-up data showed that 7 cases were fetal death, 110 women elected to terminate their pregnancies, 3 cases lost follow-up, the other 36 cases preserve continuing pregnancy, including 28 liveborn infants and 8 fetal deaths. Etiology of twin-twin transfusion syndrome, meconium peritonitis, congenital chylothorax, intrauterine infection, cardiac abnormalities and so on had survived fetus cases. The survival rate of typical hydrops fetalis in the present series was 3.6% (4/112). CONCLUSIONS: Ascites is the most common characteristics of sonogram in hydrops fetalis. The etiology of hydrops fetalis is extremely complex. The prognosis is associated with the etiology and hydrops subtype.

[The lung injury in postpartum and non-pregnant rabbits after resuscitation from hemorrhagic shock].

OBJECTIVE: To investigate the difference in the development of acute lung injury after hemorrhagic shock in postpartum and non-pregnant rabbits. METHODS: Hemorrhagic shock/resuscitation was produced on 9 pregnant New-Zealand rabbits postpartum (within 24 hours after giving birth) and 9 non-pregnant controls via carotid artery bleeding and perfusion (i.v.) of lactate Ringer solution for 3 hours. The serum level of cytokine tumor necrosis factor-α (TNF-α) and interleukin-10 (IL-10) in the animals were measured at different time points. Lung tissue were taken 3 hours after resuscitation for the determination of malondialdehyde (MDA),the activity of superoxide dismutase (SOD), myeloperoxidase (MPO), dry/wet weight (D/W), nuclear factor-ΚB (NF-ΚB), and mRNA content of intercellular adhesion molecule-1 (ICAM-1). RESULTS: Serum TNF-α (ng/L) and IL-10 (ng/L) in antepartum pregnant rabbits were not significantly different from non-pregnant rabbits (TNF-α: 87.6 ± 6.8 vs. 83.2 ± 5.3; IL-10: 44.9 ± 3.9 vs. 42.7 ± 3.4, both P > 0.05). However, the serum TNF-α increased significantly after delivery (102.5 ± 8.1 vs. 87.6 ± 6.8, P < 0.05). TNF-α and IL-10 increased 1 hour after shock in both postpartum and non-pregnant rabbits. The serum TNF-α in postpartum rabbits was significant higher than non-pregnant rabbits in each resuscitation period (1 hour: 230.0 ± 14.9 vs. 202.0 ± 10.1, 2.5 hours: 290.0 ± 18.6 vs. 236.0 ± 14.4, 4 hours: 265.0 ± 15.9 vs. 217.0 ± 12.8, all P < 0.05), meanwhile the serum IL-10 in postpartum animals was significantly lower than the non-pregnant controls(1 hour: 104.3 ± 6.9 vs. 135.0 ± 7.8, 2.5 hours: 146.8 ± 9.4 vs. 178.3 ± 11.7, 4 hours: 126.0 ± 7.9 vs. 165.8 ± 9.6, all P < 0.05). The value of MDA, MPO, D/W, NF-ΚB activity and ICAM-1 mRNA content in lung tissue of postpartum rabbits were all significant higher than non-pregnant rabbits after resuscitation [MDA (nmol/mg): 52.6 ± 5.9 vs. 39.4 ± 4.7, MPO (U/mg): 4.62 ± 0.85 vs. 3.26 ± 0.62, D/W: 0.186 ± 0.025 vs. 0.143 ± 0.016, NF-ΚB activity (A value): 0.89 ± 0.27 vs. 0.46 ± 0.15, ICAM-1 mRNA: 4.6 ± 1.2 vs. 2.5 ± 0.7, all P < 0.05], but the activity of SOD (U/mg) was lower (47.8 ± 6.7 vs. 63.5 ± 8.2, P < 0.05) in the controls after resuscitation. CONCLUSIONS: Delivery causes significant increase in serum TNF-α in pregnant rabbits. Inflammatory lung injury is more severe in postpartum rabbits after resuscitation from hemorrhagic shock, and the increased serum level of inflammatory mediators may be part of the mechanism for such difference.

Prenatal diagnosis of Pfeiffer syndrome type 2 with increased nuchal translucency.

Pfeiffer syndrome (PS) is a rare autosomal dominant genetic disorder characterized by craniosynostosis, broad thumbs / toes. Here, we report a case of PS type 2 with increased nuchal translucency in early trimester.

Identification of copy number variants by NGS-based NIPT at low sequencing depth.

OBJECTIVE: To explore the clinical utility of detecting chromosome copy number variants (CNVs) in the fetus by noninvasive prenatal testing (NIPT) using the low-pass whole-genome sequencing. METHODS: Eight hundred and seventy-three singleton pregnancies with chromosomal microarray analysis (CMA) available between January 2017 to December 2019 and stored enough plasma sample for NIPT testing were included in this study. The CMA results show that forty-eight pregnancies with CNVs and eight hundred and twenty-five pregnancies are normal. Each pregnancy's plasma sample was blindly tested with NIPT at a depth of 0.51-1.19x for CNVs detection. The performance of the NIPT method for CNVs detection compared with the CMA method is evaluated. RESULTS: A total of fifty-two CNVs ranging from 0.1-47.3 Mb identified in forty-eight samples were identified by NIPT, of which thirty-four CNVs were consistent with CMA results. Additionally, eighteen CNVs were missed by NIPT. The overall sensitivity and specificity for the detection of CNVs were 65.38% (95% CI: 51.76%-76.89%) and 97.45% (95% CI: 96.12%-98.35%), respectively. However, for the detection of CNVs larger than 2 Mb and CNVs less than 2Mb, the sensitivities were 81.58% (95% CI: 66.27%-91.09%) and 21.43% (95% CI: 6.84%-48.32%), respectively. CONCLUSION: Our study demonstrated that the NIPT might be an alternative method for screening CNVs comparable with other studies. However, CNVs less than 2Mb in length shows poor sensitivity by NIPT. Noninvasive CNVs detection based on the NIPT method still needs more clinical validation studies and technical improvement to achieve clinically acceptable accuracy.