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.

Single-cell multi-omics sequencing reveals chromosome copy number inconsistency between trophectoderm and inner cell mass in human reconstituted embryos after spindle transfer.

STUDY QUESTION: Is the chromosome copy number of the trophectoderm (TE) of a human reconstituted embryos after spindle transfer (ST) representative of the inner cell mass (ICM)? SUMMARY ANSWER: Single-cell multi-omics sequencing revealed that ST blastocysts have a higher proportion of cell lineages exhibiting intermediate mosaicism than conventional ICSI blastocysts, and that the TE of ST blastocysts does not represent the chromosome copy number of ICM. WHAT IS KNOWN ALREADY: Preimplantation genetic testing for aneuploidy (PGT-A) assumes that TE biopsies are representative of the ICM, but the TE and ICM originate from different cell lineages, and concordance between TE and ICM is not well-studied, especially in ST embryos. STUDY DESIGN, SIZE, DURATION: We recruited 30 infertile women who received treatment at our clinic and obtained 45 usable blastocysts (22 from conventional ICSI and 23 reconstituted embryos after ST). We performed single-cell multi-omics sequencing on all blastocysts to predict and verify copy number variations (CNVs) in each cell. We determined the chromosome copy number of each embryo by analysing the proportion of abnormal cells in each blastocyst. We used the Bland-Altman concordance and the Kappa test to evaluate the concordance between TE and ICM in the both groups. PARTICIPANTS/MATERIALS, SETTING, METHODS: The study was conducted at a public tertiary hospital in China, where all the embryo operations, including oocytes retrieval, ST, and ICSI, were performed in the embryo laboratory. We utilized single-cell multi-omics sequencing technology at the Biomedical Pioneering Innovation Center, School of Life Sciences, Peking University, to analyse the blastocysts. Transcriptome sequencing was used to predict the CNV of each cell through bioinformatics analysis, and the results were validated using the DNA methylation library of each cell to confirm chromosomal normalcy. We conducted statistical analysis and graphical plotting using R 4.2.1, SPSS 27, and GraphPad Prism 9.3. MAIN RESULTS AND THE ROLE OF CHANCE: Mean age of the volunteers, the blastocyst morphology, and the developmental ratewere similar in ST and ICSI groups. The blastocysts in the ST group had some additional chromosomal types that were prone to variations beyond those enriched in the blastocysts of the ICSI group. Finally, both Bland-Altman concordance test and kappa concordancetest showed good chromosomal concordance between TE and ICM in the ICSI blastocysts (kappa = 0.659, P < 0.05), but not in ST blastocysts (P = 1.000), suggesting that the TE in reconstituted embryos is not representative of ICM. Gene functional annotation (GO and KEGG analyses) suggests that there may be new or additional pathways for CNV generation in ST embryos compared to ICSI embryos. LIMITATIONS, REASONS FOR CAUTION: This study was mainly limited by the small sample size and the limitations of single-cell multi-omics sequencing technology. To select eligible single cells, some cells of the embryos were eliminated or not labelled, resulting in a loss of information about them. The findings of this study are innovative and exploratory. A larger sample size of human embryos (especially ST embryos) and more accurate molecular genetics techniques for detecting CNV in single cells are needed to validate our results. WIDER IMPLICATIONS OF THE FINDINGS: Our study justifies the routine clinical use of PGT-A in ICSI blastocysts, as we found that the TE is a good substitute for ICM in predicting chromosomal abnormalities. While PGT-A is not entirely accurate, our data demonstrate good clinical feasibility. This trial was able to provide correct genetic counselling to patients regarding the reliability of PGT-A. Regarding ST blastocysts, the increased mosaicism rate and the inability of the TE to represent the chromosomal copy number of the ICM are both biological characteristics that differentiate them from ICSI blastocysts. Currently, ST is not used clinically on a large scale to produce blastocysts. However, if ST becomes more widely used in the future, our study will be the first to demonstrate that the use of PGT-A in ST blastocysts may not be as accurate as PGT-A for ICSI blastocysts. STUDY FUNDING/COMPETING INTEREST(S): This study was supported by grants from the National Key R&D Program of China (2018YFA0107601) and the National Key R&D Program of China (2018YFC1003003). The authors declare no conflict of interest. TRIAL REGISTRATION NUMBER: N/A.

A Dual-Optimization Fault Diagnosis Method for Rolling Bearings Based on Hierarchical Slope Entropy and SVM Synergized with Shark Optimization Algorithm.

Slope entropy (SlopEn) has been widely applied in fault diagnosis and has exhibited excellent performance, while SlopEn suffers from the problem of threshold selection. Aiming to further enhance the identifying capability of SlopEn in fault diagnosis, on the basis of SlopEn, the concept of hierarchy is introduced, and a new complexity feature, namely hierarchical slope entropy (HSlopEn), is proposed. Meanwhile, to address the problems of the threshold selection of HSlopEn and a support vector machine (SVM), the white shark optimizer (WSO) is applied to optimize both HSlopEn and an SVM, and WSO-HSlopEn and WSO-SVM are proposed, respectively. Then, a dual-optimization fault diagnosis method for rolling bearings based on WSO-HSlopEn and WSO-SVM is put forward. We conducted measured experiments on single- and multi-feature scenarios, and the experimental results demonstrated that whether single-feature or multi-feature, the WSO-HSlopEn and WSO-SVM fault diagnosis method has the highest recognition rate compared to other hierarchical entropies; moreover, under multi-features, the recognition rates are all higher than 97.5%, and the more features we select, the better the recognition effect. When five nodes are selected, the highest recognition rate reaches 100%.

Long non-coding RNA HNF1A-AS1 upregulates OTX1 to enhance angiogenesis in colon cancer via the binding of transcription factor PBX3.

Colon cancer shows characteristics of metastasis, which is associated with angiogenesis. Increasing evidence highlights long non-coding RNAs (lncRNAs) as important participants in angiogenesis of cancers, including colon cancer. Hence, this study investigated the role of HNF1A-AS1 in angiogenesis of colon cancer. RT-qPCR and Western blot analysis were applied to detect HNF1A-AS1 and OTX1 expression in colon cancer tissues and cell lines. Then the interactions among HNF1A-AS1, PBX3, OTX1 and ERK/MAPK pathway were evaluated with RNA pull-down, RIP, ChIP and dual-luciferase reporter gene assays. Next, HCT116 and SW620 cells were treated with si-HNF1A-AS1 and/or oe-OTX1 plasmids to assess the effects of HNF1A-AS1 and OTX1 on angiogenesis, which was further evaluated in nude mice injected with SW620 cells transfected with sh-HNF1A-AS1 or sh-OTX1 lentivirus. HNF1A-AS1 and OTX1 were highly expressed in colon cancer. Silencing of HNF1A-AS1 inhibited angiogenesis of colon cancer in vivo and in vitro. HNF1A-AS1 increased the OTX1 expression by binding to transcription factor PBX3 to promote angiogenesis in colon cancer. Further, HNF1A-AS1 upregulated OTX1 to activate the ERK/MAPK pathway. Altogether, our findings identified HNF1A-AS1 as a tumor-promoting RNA in colon cancer, which could serve as a potential therapeutic target for colon cancer treatment.

Long non-coding RNA LINC00858 inhibits colon cancer cell apoptosis, autophagy, and senescence by activating WNK2 promoter methylation.

Accumulating evidence shows the involvement of long non-coding RNAs (lncRNAs) in tumorigenesis of many types of human cancers. However, the role of LINC00858 in colon cancer has not been fully elucidated. Therefore, we investigated the involvement of LINC00858 in the progression of colon cancer and identified its downstream targets. After examining the expression of LINC00858 in colon cancer tissues and cell lines, we then identified the possible interaction between LINC00858 and WNK lysine deficient protein kinase 2 (WNK2) by fluorescence in situ hybridization, RNA immunoprecipitation, chromatin immunoprecipitation, and RNA pull-down assays. Next, the role of the LINC00858/WNK2 axis was explored by evaluating the apoptosis, autophagy, and senescence of colon cancer cells in vitro after ectopic expression and depletion experiments in HCT116 cells. Moreover, a mouse xenograft model of HCT116 cells was established to verify the function of the LINC00858/WNK2 axis in vivo. There was high expression of LINC00858 and low expression of WNK2 in colon cancer tissues and cell lines. Silencing of LINC00858 promoted apoptosis, senescence, and autophagy in colon cancer cells. Additionally, the enrichment of WNK2 was promoted when LINC00858 bound to DNA methyltransferases. Furthermore, in vivo assays demonstrated that silencing of LINC00858 resulted in inhibited tumor growth by upregulating WNK2. In summary, LINC00858 acts as a tumor-promoting lncRNA in colon cancer by downregulating WNK2. Our results may provide novel targets for the treatment for colon cancer.

Capturing Electrocardiogram Signals from Chairs by Multiple Capacitively Coupled Unipolar Electrodes.

A prototype of an electrocardiogram (ECG) signal acquisition system with multiple unipolar capacitively coupled electrodes is designed and experimentally tested. Capacitively coupled electrodes made of a standard printed circuit board (PCB) are used as the sensing electrodes. Different from the conventional measurement schematics, where one single lead ECG signal is acquired from a pair of sensing electrodes, the sensing electrodes in our approaches operate in a unipolar mode, i.e., the biopotential signals picked up by each sensing electrodes are amplified and sampled separately. Four unipolar electrodes are mounted on the backrest of a regular chair and therefore four channel of signals containing ECG information are sampled and processed. It is found that the qualities of ECG signal contained in the four channel are different from each other. In order to pick up the ECG signal, an index for quality evaluation, as well as for aggregation of multiple signals, is proposed based on phase space reconstruction. Experimental tests are carried out while subjects sitting on the chair and clothed. The results indicate that the ECG signals can be reliably obtained in such a unipolar way.

MicroRNA-128 acts as a suppressor in the progression of gastrointestinal stromal tumor by targeting B-lymphoma Mo-MLV insertion region 1.

INTRODUCTION: The critical role of microRNA-128 (miR-128) in gastrointestinal-related diseases has been documented. In the current study, we tried to clarify the specific role miR-128 in gastrointestinal stromal tumor (GIST) and the underlying mechanism. METHODS: Differentially expressed genes in GIST were identified following bioinformatics analysis. Then, expression patterns of miR-128 and B-lymphoma Mo-MLV insertion region 1 (BMI-1) in clinical tissue samples and cell lines were characterized, followed by validation of their correlation. GIST-T1 cells were selected and transfected with different mimic, inhibitor, or siRNA plasmids, after which the biological functions were assayed. RESULTS: We identified low miR-128 and high BMI-1 expression in GIST tissues of 78 patients and 4 GIST cell lines. Ectopic expression of miR-128 or silencing of BMI-1 suppressed the malignant potentials of GIST-T1 cells. As a target of miR-128, BMI-1 re-expression could partly counteract the suppressive effect of miR-128 on the malignancy of GIST-T1 cells. CONCLUSION: Our study provided evidence that miR-128-mediated silencing of BMI-1 could prevent malignant progression of GIST, highlighting a promising anti-tumor target for combating GIST.

Integrative analysis of transcriptome, DNA methylome and chromatin accessibility reveals candidate therapeutic targets in hypertrophic cardiomyopathy.

Hypertrophic cardiomyopathy (HCM) is the most common inherited heart disease and is characterized by primary left ventricular hypertrophy usually caused by mutations in sarcomere genes. The mechanism underlying cardiac remodeling in HCM remains incompletely understood. An investigation of HCM through integrative analysis at multi-omics levels will be helpful for treating HCM. DNA methylation and chromatin accessibility, as well as gene expression, were assessed by nucleosome occupancy and methylome sequencing (NOMe-seq) and RNA-seq, respectively, using the cardiac tissues of HCM patients. Compared with those of the controls, the transcriptome, DNA methylome and chromatin accessibility of the HCM myocardium showed multifaceted differences. At the transcriptome level, HCM hearts returned to the fetal gene program through decreased sarcomeric and metabolic gene expression and increased extracellular matrix gene expression. In the DNA methylome, hypermethylated and hypomethylated differentially methylated regions (DMRs) were identified in HCM. At the chromatin accessibility level, HCM hearts showed changes in different genome elements. Several transcription factors (TFs), including SP1 and EGR1, exhibited a fetal-like pattern of binding motifs in nucleosome-depleted regions (NDRs) in HCM. In particular, the inhibition of SP1 or EGR1 in an HCM mouse model harboring sarcomere mutations markedly alleviated the HCM phenotype of the mutant mice and reversed fetal gene reprogramming. Overall, this study not only provides a high precision multi-omics map of HCM heart tissue but also sheds light on the therapeutic strategy by intervening the fetal gene reprogramming in HCM.

Chaos Raman distributed optical fiber sensing.

The physics principle of pulse flight positioning is the main theoretical bottleneck that restricts the spatial resolution of the existing Raman distributed optical fiber sensing scheme. Owing to the pulse width of tens of nanoseconds, the spatial resolution of the existing Raman distributed optical fiber sensing scheme with kilometer-level sensing distance is limited to the meter level, which seriously restricts the development of the optical time-domain reflection system. In this paper, a chaos laser is proposed in the context of the physical principle of the Raman scattering effect, and a novel theory of chaos Raman distributed optical fiber sensing scheme is presented. The scheme reveals the characteristics of chaos Raman scattering light excited by a chaotic signal on the sensing fiber. Further, the chaos time-domain compression demodulation mechanism between the temperature variation information and chaos correlation peak is demonstrated. Then, the position of the temperature variation signal is precisely located using the delay time of the chaos correlation peak combined with the chaos pulse flight time. Based on this novel optical sensing mechanism, an experiment with 10 cm spatial resolution and 1.4 km sensing distance was conducted, and the spatial resolution was found to be independent of the sensing distance. Within the limit of the existing spatial resolution theory, the spatial resolution of the proposed scheme is 50 times higher than that of the traditional scheme. The scheme also provides a new research direction for optical chaos and optical fiber sensing.

Self-management in patients with coronary heart disease after stent implantation at the long-term stage: a cross-sectional study.

BACKGROUND: The self-management of risk reduction in coronary heart disease (CHD) plays an important role in mediating health outcomes following percutaneous coronary intervention (PCI), but there is a lack of research on self-management status in the long-term stage after PCI in Chinese patients with CHD. Hence, this study investigated the self-management status in the long-term stage (>2 years) after PCI in patients with CHD. The results could provide a reference for the development of targeted interventions. METHODS: This cross-sectional study administered a questionnaire survey on self-management in patients with CHD who underwent PCI (convenience sampling) and had been discharged from our medical center for >2 years, excluding the patients with severe hepatic or renal dysfunction or tumor. Data about cardiovascular risk factors, including body mass index (BMI), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), and self-management status, were collected. The Coronary Artery Disease Self-Management Scale (CSMS) was used to assess patient self-management. RESULTS: The total CSMS score was 69.5±11.0, suggesting that the patients' self-management level was average. The scores for life management, emotion management, and disease management were 87.4±9.7, 77.6±7.7, and 57.5±11.0, respectively. Significant differences were noted in the patients' self-management scores according to occupation, education level, residence, and sex (all P<0.05). The self-management ability of farmers, primary school-educated, and male participants was relatively low. Moreover, cardiovascular risk factors were positively correlated with the emotion and disease management scores. CONCLUSIONS: CHD patients' self-management status in the long-term stage after PCI was moderate or poor. Medical staff should provide targeted guidance and education (for example, effective guidance on smoking cessation, first aid training, repeated disease education, and use of electronic devices to improve medication compliance) to improve the self-management level for the secondary prevention of CHD.

Relationship between thrombomodulin gene polymorphism and susceptibility to venous thromboembolism: A protocol for systematic review and meta-analysis.

BACKGROUND: Previous studies displayed that thrombomodulin gene polymorphisms are closely associated with venous thromboembolism (VTE), while the results are inconsistent. Therefore, we conducted a meta-analysis to accurately determine the association between thrombomodulin gene polymorphism and the risk of VTE. METHODS: Wanfang, Chinese Biomedical Literature Database, Chinese National Knowledge Infrastructure, the Chongqing VIP Chinese Science and Technology Periodical Database, PubMed, EmBase, and Web of Science databases were searched, and the time to build the database was set until January 2021. The association between thrombomodulin gene polymorphism and the risk of VTE was evaluated. Meta-analysis was performed with STATA 16.0 software, and the odds ratio and its 95% confidence interval were applied to estimate the relationship between thrombomodulin gene polym'orphism and the risk of VTE. RESULTS: The results of this meta-analysis will be submitted to a peer-reviewed journal for publication. CONCLUSION: This meta-analysis will summarize the relationship between thrombomodulin genepolymorphism and VTE risk. ETHICS AND DISSEMINATION: Ethical approval was not required for this study. The systematic review will be published in a peer-reviewed journal, presented at conferences, and shared on social media platforms. This review would be disseminated in a peer-reviewed journal or conference presentations. OSF REGISTRATION NUMBER: DOI 10.17605/OSF.IO/UEHJP.

Electronics of a Wearable ECG With Level Crossing Sampling and Human Body Communication.

In this paper, the human body communication (HBC) and level crossing sampling (LCS) are combined to design electronics for a wearable electrocardiograph (ECG). The ECG signals acquired by capacitively coupled electrodes are sampled with LCS in place of conventional synchronous sampling. In order to transmit signals through HBC at low frequencies (100 kHz, 1 MHz), an electric field sensor with high input impedance is adopted as the front end of the HBC receiver. The HBC channel gain is enhanced by more than 30 dB with the electric field sensor. An LCS structure based on the send-on-delta concept is implemented with discrete components to convert the ECG signals into binary impulses. The converted impulses are modulated by an on-off keying modulator and then transmitted via the human body to the receiver. A prototype ECG waist belt is developed with commercially available components and experimentally evaluated. The results indicate that the acquired ECG waveforms exhibit good agreement with regular Ag/AgCl ECG methods. The heartbeat detection using a technique based on the Kadane's algorithm and the power consumption performance of the proposed system are also discussed.