Single-cell multiomics analyses of spindle-transferred human embryos suggest a mostly normal embryonic development.

Mitochondrial DNA (mtDNA) mutations are often associated with incurable diseases and lead to detectable pathogenic variants in 1 out of 200 babies. Uncoupling of the inheritance of mtDNA and the nuclear genome by spindle transfer (ST) can potentially prevent the transmission of mtDNA mutations from mother to offspring. However, no well-established studies have critically assessed the safety of this technique. Here, using single-cell triple omics sequencing method, we systematically analyzed the genome (copy number variation), DNA methylome, and transcriptome of ST and control blastocysts. The results showed that, compared to that in control embryos, the percentage of aneuploid cells in ST embryos did not significantly change. The epiblast, primitive endoderm, and trophectoderm (TE) of ST blastocysts presented RNA expression profiles that were comparable to those of control blastocysts. However, the DNA demethylation process in TE cells of ST blastocysts was slightly slower than that in the control blastocysts. Collectively, our results suggest that ST seems generally safe for embryonic development, with a relatively minor delay in the DNA demethylation process at the blastocyst stage.

Personalized embryo transfer based on RNA sequencing endometrial receptivity test in repeated implantation failure patients: artificial cycle versus natural cycle.

INTRODUCTION: Embryo implantation provides an efficient way for patients with repeated implantation failure (RIF) to achieve pregnancy. The aim of this study is to compare the implantation outcomes of RIF patients in artificial cycle to those in natural cycle, both were treated with RNA sequencing endometrial receptivity test (rsERT) based personalized embryo implantation. METHODS: The endometrial receptivity (ER) analysis was performed using rsERT followed by personalized embryo transfer at optimal window of implantation (WOI). The implantation rate (IR), clinical pregnancy rate (CPR) and live birth rate (LBR) were calculated. The expression levels of biomarkers involved in pregnancy process in the patients detected as in receptivity status were also analyzed. RESULTS: The rsERT shown that 44.8% (natural cycle) and 47.8% (artificial cycle) patients were in non-receptive status, which indicated a WOI displacement. After personalized embryo transfer, the IR of patients in artificial cycle was higher than those in natural cycle (52.2% vs 27.6%). The expressions of FKBP52, MUC1 and LPAR3 were significantly lower in artificial cycle than in natural cycle. CONCLUSION: Using artificial cycle for personalized embryo transfer based on rsERT may yield better pregnancy outcomes for RIF patients. A gene expression analysis of FKBP52, MUC1 and LPAR3 provided a potential way to increase implantation outcomes for RIF patients.

Comprehensive chromosomal and mitochondrial copy number profiling in human IVF embryos.

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 (ß = 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.

Anterior uterine incarceration complicated by placenta previa and placenta accreta spectrum disorder: a case report.

Background: Uterine incarceration is a rare obstetric complication that is always associated with retroversion and prone to misdiagnosis. Pelvic examination and imaging methods including ultrasound and magnetic resonance imaging (MRI) are used as the primary diagnostic tool. We present an asymptomatic anterior uterine incarceration complicated by placenta previa and placenta accreta spectrum (PAS) disorder, which could be diagnosed during the pregnancy periods, but was first diagnosed during the cesarean section (CS) and got the surgeons into trouble. Case Description: A 28-year-old woman, gravidity 4, parity 1, was hospitalized due to placenta previa and PAS disorder diagnosed by ultrasound and MRI at 35.6 weeks of gestation. She had not experienced any discomfort. Given her history of a previous CS, she underwent a well-prepared cesarean delivery for the termination of the pregnancy. The patient had a series of periodical ultrasound and MRI examinations in which placenta previa and placenta accreta were described in disregard of the abnormal location of cervix; consequently, interior uterine incarceration was first diagnosed during the surgery, which caused significant difficulties in the operation which lasted 3 hours and 21 minutes. The patient developed severe hemorrhaging and lost approximately 5,000 mL of blood. Fortunately, she delivered a health male infant weighing 3,440 grams with quite good Apgar scores. During the follow-up, maternal and child health was confirmed. Conclusions: With regard to patients who have undergone previous pelvic surgery, doctors need to pay close attention to the position of the cervix, the pelvic adhesion situation during the prenatal examination, and be on alert for uterine incarceration. Uterine incarceration can be accurately recognized by periodic sonography and MRI if the radiologist is aware of this unusual condition. We do suggest that special are must be taken to avoid unnecessary trauma by misdiagnosis because of lack of awareness.

Long-term vaccine efficacy of a 2-dose varicella vaccine in China from 2011 to 2021: A retrospective observational study.

Objective: A 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. Methods: A 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). Results: The 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%). Conclusions: This 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.

Efficacy of low-dose hCG on FET cycle in patients with recurrent implantation failure.

Objective: To study patients' new treatment methods and mechanisms of repeated implantation failure. Design: A retrospective study. Setting: In vitro fertilization (IVF) unit in a Three-A hospital. Patients: Ninety-three patients with repeated implantation failure in IVF and embryo transfer. Interventions: the luteal phase support. Main outcome measures: According to whether human chorionic gonadotropin(HCG) was added, the two groups were divided into an observation group and a control group, and the clinical outcomes of the two groups were compared. Furthermore, 20 patients were selected for whole exome sequencing to investigate the mechanism. Results: The observation group's clinical pregnancy rate and live birth rate were significantly higher than those in the control group (P=0.004). Functional enrichment analysis showed that these genes were significantly enriched in embryo implantation or endometrial receptivity processes, such as microtubule-based movement, NABA CORE MATRISOME, superoxide anion generation, protein localization to vacuole, extracellular matrix organization, fertilization, microtubule-based transport, cell junction organization, microtubule cytoskeleton organization. Furthermore, variants detected in these pathway genes were missense mutations that affect the protein's biological activity but do not effectuate its inactivation. Conclusions: Adding HCG in the luteal phase might improve the clinical pregnancy and live birth rates in RIF patients. The potential pathogenesis of RIF genetic level may be caused by microtubule-based movement, extracellular matrix organization, and the Superoxide Anion generation pathway.

The mining and construction of a knowledge base for gene-disease association in mitochondrial diseases.

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.

Electrofusion Stimulation Is an Independent Factor of Chromosome Abnormality in Mice Oocytes Reconstructed via Spindle Transfer.

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+.

Varicella breakthrough infection and effectiveness of 2-dose varicella vaccine in China.

BACKGROUND: 2-dose varicella vaccination has been available in Ningbo city, China since 2014 in the private sector. This study aimed to evaluate the breakthrough varicella infection rate, and to examine overall and incremental effectiveness of 2-dose varicella vaccination among Chinese children. METHODS: A retrospective investigation was done among native children born from 2008 to 2013 in active surveillance area in Ningbo, China. Between 2009 and 2016, demographic information and data on varicella vaccination were collected by Ningbo's Immunization Information System, and information of varicella infections were obtained from China Information System for Disease Control and Prevention. The logistic regression was conducted to estimate varicella vaccine effectiveness (VE). RESULT: A total of 107,324 local children were enrolled in the cohort analysis and 95.11% of these children with no varicella disease history received at least 1-dose varicella vaccine from 2009 to 2016. The total breakthrough varicella infection rate (BVR) was 0.37% for all the vaccinated children and 0.04% for 2-dose vaccination. The annual BVR ranged from 0.01% to 0.49% for 1-dose of varicella vaccine and from 0.01% to 0.02% for 2-dose. The infection rates both in the unvaccinated children and the 1-dose children were decreasing after 2-dose vaccination implemented in 2014 (Unvaccinated: P < 0.001 for trend; 1-dose: P = 0.003 for trend). The VE against all varicella was 50.3% (95% CI: 39.8%-59.0%) for 1-dose (P < 0.001) and 98.7% (95% CI: 98.1%-99.1%) for 2-dose (P < 0.001), and the incremental VE was 97.4% (95% CI: 96.2%-98.2%) compared with 1-dose (P < 0.001). CONCLUSION: The 2-dose regimen provided excellent protection to prevent all varicella, and the universal 2-dose regimen of varicella vaccine should be recommended to prevent the varicella disease among children in China.

Individualized Hormone Adjustment in the Treatment of Recurrent Spontaneous Abortions.

Our goal was to develop a safe, efficient, and practical clinical plan for successful pregnancies for patients with recurrent spontaneous miscarriages by adjustment of their hormone levels after ovulation. We treated 61 patients with recurrent miscarriages and 110 patients with two miscarriages. All patients had miscarriages before or during the 12th week of pregnancy, and unsuccessfully underwent progesterone therapy. We measured their hormone levels and administered appropriate doses of estrogen, progesterone, and luteinizing hormones to attain normal levels (respectively, 150 pg/ml, 16 ng/ml, and 6 mIU/ml). The hormone doses were reduced upon detection of fetal heart beating, and the treatment continued until the 12th week of pregnancy. The patients were followed up by phone after the child birth. In patients with recurrent miscarriages, these were prevented in 57/61 (93.44 %). In patients with two miscarriages, successful pregnancies were in 106/110 (96.4 %) patients. The vast majority of patients in both groups gave birth to healthy babies. There was only one case per each group of induced labor due to trisomy 21 (patient with a history of recurrent miscarriages) or trisomy 17 (patient with two previous miscarriages). Individualized adjustment of hormone levels after ovulation prevents miscarriages and improves the pregnancy success rates.