No abstract

Single cell whole genome sequencing helps to decipher the genome heterogeneity within a cell population and facilitates the analysis of trace amounts of genetic material, such as is found in human embryos. The mitochondrial genome, although an important part of the genetic composition of eukaryotic cells, is often neglected in single cell genome analysis. A recently developed single cell whole genome amplification method was used, known as multiple annealing and looping based amplification cycles (MALBAC-NGS), for simultaneous analysis of chromosomal and mitochondrial genomes at the single cell level. The platform was validated by a series of technical and biological replicates and used for chromosomal and mitochondrial copy number analysis in 399 in-vitro fertilized embryos from 81 couples. A positive correlation of maternal age with increased mitochondria quantity (β = 0.176, P = 0.001) was observed after adjusting for the impact of cell type. Lower numbers of mitochondria were detected in successfully implanted embryos, although the difference was not significant. It is proposed that MALBAC-NGS could potentially be used for an advanced pre-implantation genetic screening procedure with both chromosomal constitution and mitochondrial copy number being evaluated.

Mitochondrial diseases are a group of heterogeneous genetic metabolic diseases caused by mitochondrial DNA (mtDNA) or nuclear DNA (nDNA) gene mutations. Mining the gene-disease association of mitochondrial diseases is helpful for understanding the pathogenesis of mitochondrial diseases, for carrying out early clinical diagnosis for related diseases, and for formulating better treatment strategies for mitochondrial diseases. This project researched the relationship between genes and mitochondrial diseases, combined the Malacards, Genecards, and MITOMAP disease databases to mine the knowledge on mitochondrial diseases and genes, used database integration and the sequencing method of the phenolyzer tool to integrate disease-related genes from different databases, and sorted the disease-related candidate genes. Finally, we screened 531 mitochondrial related diseases, extracted 26,723 genes directly or indirectly related to mitochondria, collected 24,602 variant sites on 1474 genes, and established a mitochondrial disease knowledge base (MitDisease) with a core of genes, diseases, and variants. This knowledge base is helpful for clinicians who want to combine the results of gene testing for diagnosis, to understand the occurrence and development of mitochondrial diseases, and to develop corresponding treatment methods.

ObjectiveA 2-dose varicella vaccine immunization strategy has been implemented in many cities in China, but there is few evidence on a long-term evaluation of the efficacy of the 2-dose varicella vaccine from China. This study aims to assess the long-term vaccine efficacy of the two doses varicella vaccine and analysis of its influencing factors.MethodsA retrospective study was carried out in 837,144 children born between 2011 and 2017 in Ningbo, Easten China. The logistic regression was performed to estimate varicella vaccine effectiveness (VE).ResultsThe overall VE of 2 doses of varicella vaccine compared without the vaccine was 90.31% (89.24–91.26%), and the overall incremental VE of 2 doses of varicella vaccine compared to the 1-dose was 64.71% (59.92–68.93%). Moreover, the varicella vaccination age of the second dose and the interval between 2 doses were both associated with VE. The VE compared to that without the vaccine in children vaccinated at <4 years old was 91.22% (95%CI: 90.16–92.17%) which was higher than in children vaccinated at ≥4 years old (VE: 86.79%; 95%CI: 84.52–88.73). And the effectiveness of the vaccine was 93.60% (95%CI: 92.19–94.75%) in children with the interval of the 2 doses ≤ 24 months significantly higher than in children with the interval of ≥36 months (VE: 85.62%, 95%CI: 82.89–87.91%).ConclusionsThis study provides evidence for long-term VE of the 2-dose varicella vaccine and the better age for 2-dose vaccination and the interval between 2 doses of the vaccine in China.

Oocytes reconstructed by spindle transfer (ST) are prone to chromosome abnormality, which is speculated to be caused by mechanical interference or premature activation, the mechanism is controversial. In this study, C57BL/6N oocytes were used as the model, and electrofusion ST was performed under normal conditions, Ca2+ free, and at room temperature, respectively. The effect of enucleation and electrofusion stimulation on MPF activity, spindle morphology, γ-tubulin localization and chromosome arrangement was compared. We found that electrofusion stimulation could induce premature chromosome separation and abnormal spindle morphology and assembly by decreasing the MPF activity, leading to premature activation, and thus resulting in chromosome abnormality in oocytes reconstructed via ST. Electrofusion stimulation was an independent factor of chromosome abnormality in oocytes reconstructed via ST, and was not related to enucleation, fusion status, temperature, or Ca2+. The electrofusion stimulation number should be minimized, with no more than 2 times being appropriate. As the electrofusion stimulation number increased, several typical abnormalities in chromosome arrangement and spindle assembly occurred. Although blastocyst culture could eliminate embryos with chromosomal abnormalities, it would significantly decrease the number of normal embryos and reduce the availability of embryos. The optimum operating condition for electrofusion ST was the 37°C group without Ca2+.