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Blood test is a basic medical test item, and the content of white blood cells (WBCs) is directly related to the evaluation of the patient's health and the judgment of serious diseases. However, both manual detection method and traditional image recognition method have the problem of insufficient efficiency and accuracy. The purpose of this study is to propose a fast and accurate WBC detection method, called hyperspectral based Multi-data Faster RCNN. For hyperspectral multi-dimensional sensing information, one-dimensional (1-D) CNN and Faster RCNN were used to model and analyze spectral and image features, respectively. And then, the Multi-data Faster RCNN was built, in which the 1-D CNN network and fusion network layer were added in Faster RCNN to realize the joint detection based on spectral and image fusion features of WBCs. The hyperspectral system coupled with Multi-data Faster RCNN can effectively improve the accuracy of WBCs' recognition. The average precision (Ap) value and the F1 score of each classification of WBCs were above 0.9, and the mean average precision (mAP) even reached 0.962. The results showed that the Multi-data Faster RCNN integrated with hyperspectral imaging had great potential in the detection of WBCs. ● Hyperspectral multi-dimensional sensing information can be used as an effective basis for white blood cells detection. ● Different kinds of white blood cells contain different enzymes which result in different spectral characteristics. ● Compared with 2-D image, 1-D spectral data can achieve faster model convergence with more suitable for WBC classification. ● In WBC detection, the multi-data detection has better performance in accuracy and stability than single-data detection. [ABSTRACT FROM AUTHOR]

[Display omitted] • The detection of salted sea cucumber was realized by near infrared hyperspectral imaging and low field nuclear magnetic resonance. • Cuckoo optimization algorithm was proved to be effective in extracting salient features from the data, which was helpful to improve the accuracy of the models. • Deep learning framework was used to establish moisture content prediction models of salted sea cucumber based on single-data and multiple-data. The accurate control of moisture content (MC) during the processing of sea cucumber is beneficial to improve the taste of sea cucumber and maintain its nutritional value, which is directly related to the quality and shelf life of sea cucumber. The purpose of this study is to explore the feasibility using deep learning (DL) to realize rapid nondestructive detection of MC in salted sea cucumbers based on hyperspectral imaging (HSI) and low field nuclear magnetic resonance (LF-NMR) data. Firstly, three Cuckoo Search (CS) dimensionality reduction algorithms (Traditional-CS, Binary-CS and Chaotic-CS) were combined with DL framework respectively using HSI and LF-NMR data to establish prediction models, which proved the feasibility of DL framework in predicting the MC of sea cucumbers, and Chaotic-CS algorithm was selected as the optimal dimensionality reduction algorithm. Then, the MC visualization based on HSI and LF-NMR data was realized respectively to detect the migration and decrease of MC. Finally, using both HSI and LF-NMR data, the advantages of the models based on Fusion-net DL (FDL) framework were discussed, which showed better performance than the single-data models, with R C 2 of 0.9929, RMSE C of 0.0016, R P 2 of 0.9936 and RPD of 12.5041. In summary, the rapid nondestructive detection of MC in salted sea cucumbers could be realized by HSI and LF-NMR data based on DL framework, and the advantage of data fusion detection based on FDL framework was verified. [ABSTRACT FROM AUTHOR]

Transport of mRNA is an important biological process in all cells that sets up gradients of translated protein from the site of mRNA docking and translation. Neurons are highly polarized cells where the targeted movement of RNAs and local translation at that site have been shown to be integral to the proper functioning of the neuron. Indeed, this specialized biological function for localized RNAs in particular neurons may in part confer a selective advantage on these cells such that they “out-compete” others in the race to establish synaptic connectivity. In this mini-review we highlight some of the salient features of RNA targeting and translation in neurons. [ABSTRACT FROM AUTHOR]

Abstract: RNA interference (RNAi) is a conserved eukaryotic mechanism by which double-stranded RNA (dsRNA) triggers the sequence-specific degradation of homologous mRNAs. Recent concerns have arisen in mammalian systems about off-target effects of RNAi, as well as an interferon response. Most mammalian cells respond to long dsRNAs by inducing an antiviral response mediated by interferon that leads to general inhibition of protein synthesis and nonspecific degradation of mRNAs. Moreover, recent reports demonstrate that under certain conditions, short interfering RNAs (siRNAs, 21–25 bp) may activate the interferon system. Mouse oocytes and preimplantation embryos apparently lack this response, as potent and specific inhibition of gene expression triggered by long dsRNA is observed in these cells. In the present study, we analyzed the global pattern of gene expression by microarray analysis in transgenic mouse oocytes expressing long dsRNA and find no evidence of off-targeting. We also report that genes involved in the interferon response pathway are not expressed in mouse oocytes, even after exposure for an extended period of time to long dsRNA. [Copyright &y& Elsevier]

Abstract: Zygotic gene activation is essential for development beyond the 2-cell stage in the preimplantation mouse embryo. Based on α-amanitin-sensitive BrUTP incorporation, transcription initiates in the 1-cell embryo and a major reprogramming of gene expression driven by newly expressed genes is prominently observed during the 2-cell stage. Superimposed on genome activation is the development of a transcriptionally repressive state that is mediated at the level of chromatin structure. The identity of the genes that are expressed during the 1- and 2-cell stages, however, is poorly described, as are those genes involved in mediating the transcriptionally repressive state. Using the Affymetrix MOE430 mouse GeneChip set, we characterized the set of α-amanitin-sensitive genes expressed during the 1- and 2-cell stages, and we used Expression Analysis Systematic Explorer (EASE) and Ingenuity Pathway Analysis (IPA) to identify biological and molecular processes represented by these genes, as well as interactions among them. We find that although the 1-cell embryo is transcriptionally active, we did not detect any transcripts present on the MOE430 GeneChip set to be α-amanitin-sensitive. Thus, what the BrUTP incorporation represents remains elusive. About 17% of genes expressed in the 2-cell embryo are α-amanitin-sensitive. EASE analysis reveals that genes involved in ribosome biogenesis and assembly, protein synthesis, RNA metabolism and transcription are over-represented, suggesting that genome activation during 2-cell stage may not be as global and promiscuous as previously proposed. IPA implicated Myc and Hdac1 as candidate genes involved in genome activation and the development of the transcriptionally repressive state, respectively. [Copyright &y& Elsevier]

Objective: Using fetal goats as animal models, to establish the methodology of in utero transplantation of human hematopoeitic stem cell (HSC) under B-scan ultrasonographic guidance for prenatal therapy. Study design: Human HSC were directly injected into the peritoneal cavities of the recipient fetal goats at 45–55 days of gestation (term: 145 days) under the guidance of B-type ultrasound scan. After birth, the peripheral blood was collected for fluorescence assisted cell sorting (FACS), quantitative real-time PCR and fluorescence in situ hybridization (FISH) to detect and analyze the presence of human cells in the recipients. Results: The 32 recipients were born alive except one miscarriage. To test for the presence of human-goat chimeras, cells from 13 randomly selected transplanted goats were collected. FACS analyses showed the presence of human cells in all the transplanted goats tested. The average proportion of CD34+ cells and GPA+(glycophorin A) cells in the peripheral blood were 1.34 ± 1.10% and 2.80 ± 2.10%, respectively. No CD34+ or GPA+ cells were found in the non-transplanted goats tested. The results of the quantitative real-time PCR in three engraftment goats were 1.2 × 104, 2.9 × 104, and 3.2 × 104 copies of human GPA DNA per μg of genomic DNA. FISH experiments showed that cells containing human specific α-satellite DNA sequence were present in the peripheral blood of the transplanted goats. Conclusions: The method described herein is safe and reliable, with low miscarriage risk and high chimerism rate. This approach may provide a promising animal model for potential prenatal treatment. [Copyright &y& Elsevier]

Studies using low-resolution methods to assess gene expression during preimplantation mouse development indicate that changes in gene expression either precede or occur concomitantly with the major morphological transitions, that is, conversion of the oocyte to totipotent 2-cell blastomeres, compaction, and blastocyst formation. Using microarrays, we characterized global changes in gene expression and used Expression Analysis Systematic Explorer (EASE) to identify biological and molecular processes that accompany and likely underlie these transitions. The analysis confirmed previously described processes or events, but more important, EASE revealed new insights. Response to DNA damage and DNA repair genes are overrepresented in the oocyte compared to 1-cell through blastocyst stages and may reflect the oocyte''s response to selective pressures to insure genomic integrity; fertilization results in changes in the transcript profile in the 1-cell embryo that are far greater than previously recognized; and genome activation during 2-cell stage may not be as global and promiscuous as previously proposed, but rather far more selective, with genes involved in transcription and RNA processing being preferentially expressed. These results validate this hypothesis-generating approach by identifying genes involved in critical biological processes that can be the subject of a more traditional hypothesis-driven approach. [Copyright &y& Elsevier]

An established technology to create cloned animals is through the use of somatic cell nuclear transfer (SCNT), in which reprogramming the somatic cell nucleus to a totipotent state by enucleated oocyte cytoplasm is a necessary process, including telomere length reprogramming. The limitation of this technology; however, is that the live birth rate of offspring produced through SCNT is significantly lower than that of IVF. Whether and how telomere length play a role in the development of cloned animals is not well understood. Only a few studies have evaluated this association in cloned mice, and fewer still in cloned cows. In this study, we investigated the difference in telomere length as well as the abundance of some selected molecules between newborn deceased cloned calves and normal cows of different ages either produced by SCNT or via natural conception, in order to evaluate the association between telomere length and abnormal development of cloned cows. The absolute telomere length and relative mitochondrial DNA (mtDNA) copy number were determined by real-time quantitative PCR (qPCR), telomere related gene abundance by reverse-transcription quantitative PCR (RT-qPCR), and senescence-associated β-galactosidase (SA-β-gal) expression by SA-β-gal staining. The results demonstrate that the newborn deceased SCNT calves had significantly shortened telomere lengths compared to newborn naturally conceived calves and newborn normal SCNT calves. Significantly lower mtDNA copy number, and significantly lower relative abundance of LMNB1 and TERT , higher relative abundance of CDKN1A , and aberrant SA-β-gal expression were observed in the newborn deceased SCNT calves, consistent with the change in telomere length. These results demonstrate that abnormal telomere shortening, lower mtDNA copy number and abnormal abundance of related genes were specific to newborn deceased SCNT calves, suggesting that abnormally short telomere length may be associated with abnormal development in the cloned calves. • The association of telomere length and related genes with developmental abnormalities in cloned calves was investigated. • Shortened telomere length and reduced mtDNA copy number were specific to newborn deceased SCNT calves. • Abnormal telomere shortening may be associated with abnormal development in cloned cows. • To help further understand the role of telomeres in developmental barriers to cloned calves. • Telomeres and related genes may be able to serve as molecular targets to enhance live birth rates in cloned calves. [ABSTRACT FROM AUTHOR]

The surface of carp is easily damaged during the descaling process, which jeopardizes the quality and safety of carp products. Damage recognition realized by manual detection is an important factor restricting the automation in the pretreatment. For the commonly used methods of mechanical and water‐jet descaling, damage area recognition according to the hyperspectral data was proposed. Two discrimination models, including decision tree (DT) and self‐organizing feature mapping (SOM), were established to recognize the damaged and normal descaling area with the average spectral value. The damage‐discrimination model based on DT was determined to be the optimal one, which possessed the best model performance (accuracy = 96.7%, sensitivity = 96.7%, specificity = 96.7%, F1‐score = 96.7%). Considering the efficiency and precision of damage‐area recognition and visualization, the principal component analysis (PCA) combined with pixel values statistical analysis was used to reduce the dimension of hyperspectral images at the image level. Through statistical analysis, the value 0 was used as the threshold to distinguish the normal area and the damaged area in the PC image to achieve preliminary segmentation. Then, the spectral values of the initially discriminated damage area were input into the DT discrimination model to realize the final discriminant of damaged area. On this basis, the position information of the damaged area could be used to realize the visualization. The final visualization maps for mechanical and water‐jet descaling damage were obtained by image morphology processing. The average recognition accuracy can reach 94.9% and 90.3%, respectively. The results revealed that the hyperspectral imaging technique has great potential to recognize the carp damage area nondestructively and accurately under descaling processing. Practical Application: This study demonstrated that hyperspectral imaging technique can realize the carp damage area detection nondestructively and accurately under descaling processing. With the advantages of nondestructive and rapid, hyperspectral imaging system and the method can be widely expanded and applied to the quality detection of other freshwater fish pretreatment. [ABSTRACT FROM AUTHOR]

RNA editing is a co- or post-transcriptional modification through which some cells can make discrete changes to specific nucleotide sequences within an RNA molecule after transcription. Previous studies found that RNA editing may be critically involved in cancer and aging. However, the function of RNA editing in human early embryo development is still unclear. In this study, through analyzing single cell RNA sequencing data, 36.7% RNA editing sites were found to have a have differential editing ratio among early embryo developmental stages, and there was a great reprogramming of RNA editing rates at the 8-cell stage, at which most of the differentially edited RNA editing sites (99.2%) had a decreased RNA editing rate. In addition, RNA editing was more likely to occur on RNA splicing sites during human early embryo development. Furthermore, long non-coding RNA (lncRNA) editing sites were found more likely to be on RNA splicing sites (odds ratio = 2.19, P = 1.37×10−8), while mRNA editing sites were less likely (odds ratio = 0.22, P = 8.38×10−46). Besides, we found that the RNA editing rate on lncRNA had a significantly higher correlation coefficient with the percentage spliced index (PSI) of lncRNA exons (R = 0.75, P = 4.90×10−16), which indicated that RNA editing may regulate lncRNA splicing during human early embryo development. Finally, functional analysis revealed that those RNA editing-regulated lncRNAs were enriched in signal transduction, the regulation of transcript expression, and the transmembrane transport of mitochondrial calcium ion. Overall, our study might provide a new insight into the mechanism of RNA editing on lncRNAs in human developmental biology and common birth defects. Author summary: RNA editing is a process by which selected nucleotides on RNA molecules are changed post transcription of RNA. This results in changes RNA sequences that are different from the corresponding DNA sequence at modified positions. However, this slight difference has a great impact on various biological processes. In this manuscript, we investigated the function of RNA editing during human early embryo development. Through analyzing the sequencing data and computational biology, we found that RNA editing could regulate the alternative splicing of long-noncoding RNAs (lncRNAs) in human early embryo development. Here, lncRNAs are special kind of RNAs that do not translate into proteins, but have functions in the regulation of other genes. Alternative splicing is a process through which RNA molecules can be spliced into different transcripts after transcription. Thus, our results indicated that RNA editing could regulate the alternative splicing of lncRNAs and further affect the expression of downstream genes in human early embryo development. RNA editing could be the reason for the tremendous change of gene expression profile in human early embryo development which was found the previous researches. [ABSTRACT FROM AUTHOR]

[Display omitted] • Photothermal BLC can be directely carbonized from bamboo leaves. • The BLC/MF achieves an evaporation rate up to 2.29 kg m-2h−1 under one-sun irradaition. • The BLC/MF exhibits the excellent stability for long-term solar steam generation. • Salt tolerant desalination and wastewater purification over BLC/MF are demonstrated. High-performance salt-resistant solar evaporators breaking the theoretical limit of solar steam generation are highly desired for solar water evaporation technology. However, low-cost, scalable and continuous solar steam generation remains a big challenge. Herein, a robust biomass-based salt-resistant three-dimensional (3D) evaporator is designed and developed, which is composed of bamboo leaf-derived carbon (BLC) and heat-insulating macroporous melamine foam (MF), showing the integrated merits for the boosted solar evaporation performances. The designed BLC/MF evaporator presents excellent photothermal conversion ability with an evaporation rate as high as 2.29 kg m-2h−1 and acceptable evaporation efficiency of 93.95 % under one-sun irradiation. The superior performance endows BLC/MF evaporator with flexible and versatile behaviors to purify various water sources to produce clean water meeting the standard level of World Health Organization (WHO) for drinkable water. Significantly, the BLC/MF evaporator maintains a similar evaporation rate in saline solutions and can work stably at high salinities without salt deposition. This present discloses that the BLC/MF evaporator can be served as a feasible and robust alternative for solar water evaporation in various water environment for clean water production. [ABSTRACT FROM AUTHOR]

Inflammation and fibrosis in human liver are often precursors to hepatocellular carcinoma (HCC), yet none of them is easily modeled in animals. We previously generated transgenic mice with hepatocyte-specific expressed herpes simplex virus thymidine kinase (HSV-tk). These mice would develop hepatitis with the administration of ganciclovir (GCV). However, our HSV-tk transgenic mice developed hepatitis and HCC tumor as early as six months of age even without GCV administration. We analyzed the transcriptome of the HSV-tk HCC tumor and hepatitis tissue using microarray analysis to investigate the possible causes of HCC. Gene Ontology (GO) enrichment analysis showed that the up-regulated genes in the HCC tissue mainly include the immune-inflammatory and cell cycle genes. The down-regulated genes in HCC tumors are mainly concentrated in the regions related to lipid metabolism. Gene set enrichment analysis (GSEA) showed that immune-inflammatory-related signals in the HSV-tk mice are up-regulated compared to those in Notch mice. Our study suggests that the immune system and inflammation play an important role in HCC development in HSV-tk mice. Specifically, increased expression of immune-inflammatory-related genes is characteristic of HSV-tk mice and that inflammation-induced cell cycle activation maybe a precursory step to cancer. The HSV-tk mouse provides a suitable model for the study of the relationship between immune-inflammation and HCC, and their underlying mechanism for the development of therapeutic application in the future. [ABSTRACT FROM AUTHOR]

Hematopoietic stem cells (HSCs) are stem cells that can differentiate into various blood cells and have long-term self-renewal capacity. At present, HSC transplantation is an effective therapeutic means for many malignant hematological diseases, such as aplastic hematological diseases and autoimmune diseases. The hematopoietic microenvironment affects the proliferation, differentiation, and homeostasis of HSCs. The regulatory effect of the hematopoietic microenvironment on HSCs is complex and has not been thoroughly studied yet. In this study, we focused on mononuclear cells (MNCs), which provided an important microenvironment for HSCs and established a methodological system for identifying cellular composition by means of multiple technologies and methods. First, single-cell RNA sequencing (scRNA-seq) technology was used to investigate the cellular composition of cells originating from different microenvironments during different stages of hematopoiesis, including mouse fetal liver mononuclear cells (FL-MNCs), bone marrow mononuclear cells (BM-MNCs), and in vitro-cultured fetal liver stromal cells. Second, bioinformatics analysis showed a higher proportion and stronger proliferation of the HSCs in FL-MNCs than those in BM-MNCs. On the other hand, macrophages in in vitro-cultured fetal liver stromal cells were enriched to about 76%. Differential gene expression analysis and Gene Ontology (GO) functional enrichment analysis demonstrated that fetal liver macrophages have strong cell migration and actin skeleton formation capabilities, allowing them to participate in the hematopoietic homeostasis through endocytosis and exocytosis. Last, various validation experiments such as quantitative real-time PCR (qRT-PCR), ELISA, and confocal image assays were performed on randomly selected target genes or proteins secreted by fetal liver macrophages to further demonstrate the potential relationship between HSCs and the cells inhabiting their microenvironment. This system, which integrates multiple methods, could be used to better understand the fate of these specific cells by determining regulation mechanism of both HSCs and macrophages and could also be extended to studies in other cellular models. [ABSTRACT FROM AUTHOR]

Thalassemia is a recessive monogenic hematological disease associated with reduced amounts of functional hemoglobin caused by mutations/deletions in at least one of the globin genes. This disease has attracted significant attention throughout the years in terms of genetic diagnosis and developments in gene and cell therapy. Here, recent progress is reviewed in the genetic diagnosis and development of therapeutics for thalassemia, particularly β-thalassemia, in China and around the world. [ABSTRACT FROM AUTHOR]

Conventional silicon (Si)-based power devices face physical limitations—such as switching speed and energy efficiency—which can make it difficult to meet the increasing demand for high-power, low-loss, and fast-switching-frequency power devices in power electronic converter systems. Gallium nitride (GaN) is an excellent candidate for next-generation power devices, capable of improving the conversion efficiency of power systems owing to its wide band gap, high mobility, and high electric breakdown field. Apart from their cost effectiveness, GaN-based power high-electron-mobility transistors (HEMTs) on Si substrates exhibit excellent properties—such as low ON-resistance and fast switching—and are used primarily in power electronic applications in the fields of consumer electronics, new energy vehicles, and rail transit, amongst others. During the past decade, GaN-on-Si power HEMTs have made major breakthroughs in the development of GaN-based materials and device fabrication. However, the fabrication of GaN-based HEMTs on Si substrates faces various problems—for example, large lattice and thermal mismatches, as well as 'melt-back etching' at high temperatures between GaN and Si, and buffer/surface trapping induced leakage current and current collapse. These problems can lead to difficulties in both material growth and device fabrication. In this review, we focused on the current status and progress of GaN-on-Si power HEMTs in terms of both materials and devices. For the materials, we discuss the epitaxial growth of both a complete multilayer HEMT structure, and each functional layer of a HEMT structure on a Si substrate. For the devices, breakthroughs in critical fabrication technology and the related performances of GaN-based power HEMTs are discussed, and the latest development in GaN-based HEMTs are summarised. Based on recent progress, we speculate on the prospects for further development of GaN-based power HEMTs on Si. This review provides a comprehensive understanding of GaN-based HEMTs on Si, aiming to highlight its development in the fields of microelectronics and integrated circuit technology. [ABSTRACT FROM AUTHOR]

Limb‐girdle muscular dystrophy recessive 1 (LGMDR1), previously known as LGMD2A, is a specific LGMD caused by a gene mutation encoding the calcium‐dependent neutral cysteine protease calpain‐3 (CAPN3). In our study, the compound heterozygosity with two missense variants c.635 T > C (p.Leu212Pro) and c.2120A > G (p.Asp707Gly) was identified in patients with LGMDR1. However, the pathogenicity of c.635 T > C has not been investigated. To evaluate the effects of this novel likely pathogenic variant to the motor system, the mouse model with c.635 T > C variant was prepared by CRISPR/Cas9 gene editing technique. The pathological results revealed that a limited number of inflammatory cells infiltrated the endomyocytes of certain c.635 T > C homozygous mice at 10 months of age. Compared with wild‐type mice, motor function was not significantly impaired in Capn3 c. 635 T > C homozygous mice. Western blot and immunofluorescence assays further indicated that the expression levels of the Capn3 protein in muscle tissues of homozygous mice were similar to those of wild‐type mice. However, the arrangement and ultrastructural alterations of the mitochondria in the muscular tissues of homozygous mice were confirmed by electron microscopy. Subsequently, muscle regeneration of LGMDR1 was simulated using cardiotoxin (CTX) to induce muscle necrosis and regeneration to trigger the injury modification process. The repair of the homozygous mice was significantly worse than that of the control mice at day 15 and day 21 following treatment, the c.635 T > C variant of Capn3 exhibited a significant effect on muscle regeneration of homozygous mice and induced mitochondrial damage. RNA‐sequencing results demonstrated that the expression levels of the mitochondrial‐related functional genes were significantly downregulated in the mutant mice. Taken together, the results of the present study strongly suggested that the LGMDR1 mouse model with a novel c.635 T > C variant in the Capn3 gene was significantly dysfunctional in muscle injury repair via impairment of the mitochondrial function. [ABSTRACT FROM AUTHOR]

A novel E-mode gallium nitride (GaN) high-electron-mobility field-effect transistor (HEMT) was prepared by combining a superlattice barrier with Mg by thermal diffusion doping. The new device can effectively overcome the disadvantages of the conventional p-GaN gate E-mode HEMT process, which include high etching accuracy requirements, difficult p-type doping, weak gate control, and large gate leakage. The device obtained in this study has a threshold voltage of 1.17 V, an output current of 227 mA mm−1 at VG = 3 V, a gate leakage of 2.0 × 10−8 mA mm−1 at VG = 0 V, and a breakdown voltage of up to 405 V. The high-voltage, low-gate-leakage current power device developed in this work could be used as a new type of E-mode power device. [ABSTRACT FROM AUTHOR]

Objectives: Mesoderm, derived from a new layer between epiblast and hypoblast during gastrulation, can differentiate into various tissues, including muscles, bones, kidneys, blood, and the urogenital system. However, systematic elucidation of mesoderm characteristics and specific markers remains a challenge. This study aims to screen and identify candidate genes important for mesoderm development. Materials and Methods: Cells originating from the three germ layers were obtained by laser capture microdissection, followed by microcellular RNA sequencing. Mesoderm‐specific differentially expressed genes (DEGs) were identified by using a combination of three bioinformatics pipelines. Candidate mesoderm‐specific genes expression were verified by real‐time quantitative polymerase chain reaction analysis and immunohistochemistry. Functional analyses were verified by ESCs‐EBs differentiation and colony‐forming units (CFUs) assay. Results: A total of 1962 differentially expressed mesoderm genes were found, out of which 50 were candidate mesoderm‐specific DEGs which mainly participate in somite development, formation of the primary germ layer, segmentation, mesoderm development, and pattern specification process by GO analysis. Representative genes Cdh2, Cdh11, Jag1, T, Fn‐1, and Pcdh7 were specifically expressed in mesoderm among the three germ layers. Pcdh7 as membrane‐associated gene has hematopoietic‐relevant functions identified by ESCs‐EBs differentiation and CFUs assay. Conclusions: Spatial transcriptomic profiling with multi‐method analysis and confirmation revealed candidate mesoderm progenitors. This approach appears to be efficient and reliable and can be extended to screen and validate candidate genes in various cellular systems. [ABSTRACT FROM AUTHOR]

Pathogenic variants of zinc finger C4H2-type containing (ZC4H2) on the X chromosome cause a group of genetic diseases termed ZC4H2-associated rare disorders (ZARD), including Wieacker-Wolff Syndrome (WRWF) and Female-restricted Wieacker-Wolff Syndrome (WRWFFR). In the current study, a de novo c.352C>T (p.Gln118*) mutation in ZC4H2 (NM_018684.4) was identified in a female neonate born with severe arthrogryposis multiplex congenita (AMC) and Pierre-Robin sequence (cleft palate and micrognathia). Plasmids containing the wild-type (WT), mutant-type (MT) ZC4H2, or GFP report gene (N) were transfected in 293T cell lines, respectively. RT-qPCR and western blot analysis showed that ZC4H2 protein could not be detected in the 293T cells transfected with MT ZC4H2. The RNA seq results revealed that the expression profile of the MT group was similar to that of the N group but differed significantly from the WT group, indicating that the c.352C>T mutation resulted in the loss of function of ZC4H2. Differentially expressed genes (DEGs) enrichment analysis showed that c.352C>T mutation inhibited the expression levels of a series of genes involved in the oxidative phosphorylation pathway. Subsequently, expression levels of ZC4H2 were knocked down in neural stem cells (NSCs) derived from induced pluripotent stem cells (iPSCs) by lentiviral-expressed small hairpin RNAs (shRNAs) against ZC4H2. The results also demonstrated that decreasing the expression of ZC4H2 significantly reduced the growth of NSCs by affecting the expression of genes related to the oxidative phosphorylation signaling pathway. Taken together, our results strongly suggest that ZC4H2 c.352C>T (p.Gln118*) mutation resulted in the loss of protein function and caused WRWFFR. [ABSTRACT FROM AUTHOR]

Objectives: Early embryo development is dependent on the regulation of maternal messages stored in the oocytes during the maternal‐to‐zygote transition. Previous studies reported variability of oocyte competence among different inbred mouse strains. The present study aimed to identify the maternal transcripts responsible for early embryonic development by comparing transcriptomes from oocytes of high‐ or low‐ competence mouse strains. Materials and Methods: In vitro fertilization embryos from oocytes of different mouse strains were subject to analysis using microarrays, RNA sequencing, real‐time quantitative PCR (RT‐qPCR) analysis, Western blotting, and immunofluorescence. One candidate gene, Prkce, was analysed using Prkce knockout mice, followed by a cRNA rescue experiment. Results: The fertilization and 2‐cell rate were significantly higher for FVB/NJ (85.1% and 82.0%) and DBA/2J (79.6% and 76.7%) inbred mouse strains than those for the MRL/lpr (39.9% and 35.8%) and 129S3 (35.9% and 36.6%) strains. Thirty‐nine differentially expressed genes (DEGs) were noted, of which nine were further verified by RT‐qPCR. Prkce knockout mice showed a reduced 2‐cell rate (Prkce+/+ 80.1% vs. Prkce−/− 32.4%) that could be rescued by Prkce cRNA injection (2‐cell rate reached 76.7%). Global transcriptional analysis revealed 143 DEGs in the knockout mice, which were largely composed of genes functioning in cell cycle regulation. Conclusions: The transcription level of maternal messages such as Prkce in mature oocytes is associated with different 2‐cell rates in select inbred mouse strains. Prkce transcript levels could serve as a potential biomarker to characterize high‐quality mature oocytes. [ABSTRACT FROM AUTHOR]

Pantothenate kinase associated neurodegeneration (PKAN) is a severe autosomal recessive rare disease and characterized by iron accumulation in the basal ganglia. To investigate the pathogenesis of this disease in two sibling patients with PANK in a Chinese family, whole-exome variant detection and functional analysis were performed. Clinical and radiographic investigations were performed in the two brother patients. Whole exome sequencing (WES) was used in mutation detection, and the mutations were confirmed by Sanger sequencing. A longevity cohort genetic database was applied as Chinese urban controls. Bioinformatic analysis was performed to predict the pathogenicity. Compound heterozygous mutations of PANK2 were detected in two sibling brothers with PKAN in a Chinese family: c.510_522del (p.A170fs) and c.1319G > C (p.R440P) in the transcript NM_153638. PANK2: c.510_522del (p.A170fs) was absent in public data and the Chinese urban controls. Bioinformatics analysis showed that the above two variants were pathogenicity. We identified a rare compound heterozygous combination of PANK2 mutations found in a Chinese family in which two sibling brothers suffered from PKAN. PANK2 c.510_522del (p.A170fs) was the first reported to be a PKAN pathogenic variant. [ABSTRACT FROM AUTHOR]

We examined changes in burnout across three timepoints in one school year, in a sample (N = 230) of special educators serving students with emotional-behavioral disorders, in 15 school districts selected through stratified random sampling at the national level. Emotional exhaustion decreased at each timepoint in the school year and personal accomplishment increased from fall to spring. Latent growth curve modeling did not produce latent trajectories of burnout among teachers; however, cross-lagged panel structure equation modeling revealed that emotional exhaustion and personal accomplishment had both direct and indirect effects on depersonalization. Differences in burnout were significant by race/ethnicity but not by gender. Participants reported higher emotional exhaustion, lower depersonalization, and higher personal accomplishment than a national sample. We provide implications for researchers and practitioners. [ABSTRACT FROM AUTHOR]

Mental retardation is the main clinical manifestation of Down syndrome (DS), and neural abnormalities occur during the early embryonic period and continue throughout life. Tc1, a model mouse for DS, carries the majority part of the human chromosome 21 and has multiple neuropathy phenotypes similar to patients with DS. To explore the mechanism of early neural abnormalities of Tc1 mouse, induced pluripotent stem (iPS) cells from Tc1 mice were obtained, and genome‐wide gene expression and methylation analysis were performed for Tc1 and wild‐type iPS cells. Our results showed hypermethylation profiles for Tc1 iPS cells, and the abnormal genes were shown to be related to neurodevelopment and distributed on multiple chromosomes. In addition, important genes involved in neurogenesis and neurodevelopment were shown to be downregulated in Tc1 iPS cells. In short, our study indicated that genome‐wide hypermethylation leads to the disordered expression of genes associated with neurodevelopment in Tc1 mice during early development. Overall, our work provided a useful reference for the study of the molecular mechanism of nervous system abnormalities in DS. [ABSTRACT FROM AUTHOR]

Two-dimensional polymer/nanoparticle (NP) composite superlattice monolayers have attracted great attention, due to their applications in flexible optical, sensing, and electronic devices. The large-scale fabrication of these devices requires large-area superlattice monolayers. However, the preparation of large-area superlattice monolayers of polymer-grafted NP still faces great challenges. Herein, we report a modified Langmuir-Blodgett method for preparing large-area superlattice monolayers of polystyrene-grafted gold NPs (AuNP@PS). This method involves two key steps: pre-assembling the AuNP@PS on a liquid surface using mixed high/low-boiling-point solvent as the spreading solvent and then compressing the pre-assembled film in low-boiling-point solvent vapor. The discrete monolayer islands of AuNP@PS are formed in the pre-assembly process. During the surface compression process, the introduction of solvent vapor facilitates the fusion of AuNP@PS islands to form the continuous superlattice monolayers. Otherwise, discrete films with nonuniform thickness are obtained by compression without solvent vapor. This strategy enables the preparation of large superlattice monolayer films over 100 cm2, which are useful in the manufacture of microelectronic devices. [Display omitted] • A modified Langmuir-Blodgett method for preparing superlattice films is reported. • Superlattice films of polymer-grafted nanoparticles over 100 cm2 can be obtained. • Mixed solvent and solvent vapor facilitate the formation of superlattice monolayers. [ABSTRACT FROM AUTHOR]

Nanog is a well-known transcription factor that plays a fundamental role in stem cell self-renewal and the maintenance of their pluripotent cell identity. There remains a large data gap with respect to the spectrum of the key pluripotency transcription factors' interaction partners. Limited information is available concerning Nanog-associated RNA-binding proteins (RBPs), and the intrinsic protein-RNA interactions characteristic of the regulatory activities of Nanog. Herein, we used an improved affinity protocol to purify Nanog-interacting RBPs from mouse embryonic stem cells (ESCs), and 49 RBPs of Nanog were identified. Among them, the interaction of YBX1 and ILF3 with Nanog mRNA was further confirmed by in vitro assays, such as Western blot, RNA immunoprecipitation (RIP), and ex vivo methods, such as immunofluorescence staining and fluorescent in situ hybridization (FISH), MS2 in vivo biotin-tagged RNA affinity purification (MS2-BioTRAP). Interestingly, RNAi studies revealed that YBX1 and ILF3 positively affected the expression of Nanog and other pluripotency-related genes. Particularly, downregulation of YBX1 or ILF3 resulted in high expression of mesoderm markers. Thus, a reduction in the expression of YBX1 and ILF3 controls the expression of pluripotency-related genes in ESCs, suggesting their roles in further regulation of the pluripotent state of ESCs. [ABSTRACT FROM AUTHOR]

Streptomyces phage φC31 integrase induces efficient site-specific recombination capable of integrating exogenous genes at pseudo attP sites in human, mouse, rat, rabbit, sheep, Drosophila, and bovine genomes. However, the φC31-mediated recombination between attB and the corresponding pseudo attP sites has not been investigated in Capra hircus. Here, we identified eight pseudo attP sites located in the intron or intergenic regions of the C. hircus genome, and demonstrated different levels of foreign gene expression after φC31 integrase-mediated integration. These pseudo attP sites share similar sequences with each other and with pseudo attP sites in other mammalian genomes, and these are associated with a neighboring consensus motif found in other genomes. The application of the φC31 integrase system in C. hircus provides a new option for genetic engineering of this economically important goat species. [ABSTRACT FROM AUTHOR]

Highlights: [•] Combination of HR and ΦC31 integrase was used to improve site-specific integration. [•] ΦC31 integrase was used to introduce double strand breaks in vivo. [•] The frequency of integration into the targeted sites was evaluated up to 93%. [•] A new way for effective integration of donor gene at desired locus was provided. [ABSTRACT FROM AUTHOR]

Human amniotic fluid derived progenitor cells (hAFPCs) may be multipotent and can be considered a potential tool in the field of cell therapy for haemophilia B. Their capacity to express human coagulation factor IX (hFIX) after transduction and their fate after in utero transplantation is unknown. hAFPCs isolated from second trimester pregnancies were assessed for their phenotypic markers, multilineage capacity, and expression of hFIX after transduction. Their engraftment potential was analysed in a mouse model after in utero transplantation at embryonic day 12.5. Immunohistochemistry, fluorescence in situ, ELISA and PCR were used to assess post-transplant chimeras. hAFPCs expressed several pluripotent markers, including NANOG, SOX2, SSEA4 and TRA-1-60, and could differentiate into adipocytes and osteocytes. In vitro, after transduction with hFIX and EGFP cDNAs, constitutive hFIX protein expression and clotting activity were found. Engraftment was achieved in various foetal tissues after in utero transplantation. Safe engraftment without oncogenesis was confirmed, with low donor cell levels, but persistent engraftment, into different organs (liver, heart and lung) through to 12 weeks of age. Transgenic expression of circulating hFIX was detected in recipient mice for up to 12 weeks. hAFPCs can be engrafted long-term in immunocompetent mice after in utero transplantation. Thus, cell transplantation approaches using genetically engineered hAFPCs may prove valuable for the prenatal treatment for haemophilia B. [ABSTRACT FROM AUTHOR]

ΦC31 integrase, a site-specific recombinase, can catalyze integration of circular DNA bearing attB site into pseudo attP sites in mammalian genomes. However, the integration efficiency mediated by integrase is relatively low. Our study centered on the investigation of the impact of the position, orientation, and number of attBs in the donor plasmid on the efficiency of ΦC31 integrase system. Donor plasmids bearing various types of attBs (including forward and reverse directions, tandem, and intersperse) and reporter enhanced green fluorescent protein ( EGFP) were constructed. The plasmids plus helper plasmid encoding integrase were co-transfected into HeLa cells. After G418 selection, the resistant cell colonies were counted for calculating chromosomal integration frequency. EGFP expression was detected by fluorescence-activated cell sorter and enzyme-linked immunosorbent assay analysis. The results showed that efficiency of integration mediated by integrase accounted for 70%±7.1% of total integration events in the transfected HeLa cells. Compared with a forward orientation of attB in donor plasmid, a reverse direction of attB or interspersed attBs showed 1.5- or 2.8-fold increase in integration efficiency, respectively, while tandem attBs in donor plasmids caused a decreased efficiency of integration. We conclude that the adjustment of attB sites in donor plasmids may be of value for gene therapy and routine genetic engineering by using ΦC31 integrase system. [ABSTRACT FROM AUTHOR]

Induced pluripotent stem (iPS) cells hold great promise for regenerative medicine and drug discovery. Since the invention of iPS technology, a central goal of this field has been to derive safer and better iPS cells for human research and therapeutic applications. From the first generation of iPS cells that were only partially pluripotent, through iPS cells that were capable of germ line transmission, to current iPS cells that can produce viable mice through tetraploid complementation, the accumulating knowledge gained through mouse models is expected to extend to humans for selection and characterization of fully pluripotent human iPS cell lines. Here we review the progress and strategies toward generating fully pluripotent iPS cells, discuss the potential of patient-specific iPS cells in disease modeling and drug discovery, and discuss the potential for novel gene therapy systems combined with iPS cell-based cell replacement therapy. [ABSTRACT FROM AUTHOR]

Abstract: A novel intrauterine transplantation (IUT) approach was developed to improve the efficiency of engraftment of hematopoietic stem cells (HSCs). HSCs with a green fluorescent protein (GFP) reporter gene were transplanted in utero on days 12.5, 13.5 and 14.5 post coitum (p.c.). The degree of chimerism of donor cells in recipient newborn mice was examined using fluorescent microscopy, polymerase chain reaction (PCR), fluorescence-activated cell sorting (FACS), and fluorescence in situ hybridization (FISH) analyses. Microscopic examination revealed the presence of green fluorescent signal in the peripheral blood of the chimeric mice. The highest survival rate (47%) as well as the highest chimerism rate (73%) were achieved by our new approach in the newborn mice that were subjected to in utero transplantation (IUT) on day 12.5 p.c. (E12.5) compared to the conventional IUT method. FACS analysis indicated that 1.55±1.10% of peripheral blood cells from the newborn mice were GFP-positive donor cells. FISH showed that cells containing the donor-specific GFP sequence were present in the bone marrow (BM) of the chimeric mice. Thus, the efficiency of chimera production with this new method of IUT was significantly improved over the existing IUT techniques and instruments. [Copyright &y& Elsevier]

Salted (S) sea cucumbers are not only used as the ingredient in most sea cucumber products but also sold directly as the final product. Over-salted (OS) and sugar-treated (ST) sea cucumbers are usually used to adulterate salted sea cucumbers, which jeopardizes the quality and safety of salted sea cucumber products. In this study, S, OS and ST sea cucumbers were classified according to the hyperspectral data in the near-infrared range (957.3–1679.8 nm). Four preprocessing methods, including Savitzky–Golay (SG), multiplicative scatter correction (MSC), standard normal variate (SVN) and variable sorting for normalization (VSN) were established to filter the noise and scattering information in the original spectra. The approaches of regression coefficient (RC), successive projections algorithm (SPA), and two-dimensional correlation spectroscopy (2D-COS) were adopted to extract characteristic wavelengths. Subsequently, sea cucumber classification models were established on the basis of full and characteristic spectra using a support vector machine (SVM). Among the models established, the SNV-2D-COS-SVM and VSN-2D-COS-SVM classification models showed optimal recognition performance. In summary, the rapid and non-destructive classification and identification of S, OS, and ST sea cucumbers were accurately realized with the near-infrared-based HSI technology, and the feasibility of using near-infrared hyperspectral technology to identify salted sea cucumber was verified. • Hyperspectral imaging was applied for salted sea cucumbers adulteration detection. • Spectral data processing is helpful to improve the accuracy of the model. • The classification model of sea cucumber was established by support vector machine. [ABSTRACT FROM AUTHOR]

Abstract: The aim of this study was to determine the effect of individual oocyte donors on cloned embryo development in vitro. Five Holstein heifers of varied genetic origins were subject to ovum pick up (OPU) once weekly. In total, 913 oocytes were recovered from 1304 follicles. A mean of 7.7±0.4 oocytes was recovered per session per animal. Individual mean oocyte production varied significantly in quantity but not in quality (morphological categories) among heifers. Oocytes from individual heifers were used as recipient cytoplasm for somatic cell nuclear transfer (SCNT). Cumulus cells, collected from a single Holstein cow genetically unrelated to the oocyte donor, were used as donor cells. Although the percentage of reconstructed embryos that started to cleave was nearly constant, the percentage of cleaved embryos that developed into blastocysts showed clear individual heifer variation (61%, 51%, 31%, 28% and 24%, respectively), with a mean of 38% showing blastocyst formation. In vitro fertilization (IVF) was also conducted with oocyte from the same heifers used in SCNT. A variation of blastocyst production among individual heifers was also shown in the IVF experiment, but the rank of oocyte donor based on the blastocyst rate was changed. In conclusion, individual oocyte donor may have an effect on cloned embryo development in vitro, which differed from the effect on IVF embryos. [Copyright &y& Elsevier]

Novel methods and associated tools permitting individual identification in publicly accessible SNP databases have become a debatable issue. There is growing concern that current technical and ethical safeguards to protect the identities of donors could be insufficient. In the context of human embryonic stem cell research, there are no studies focusing on the probability that an hESC line donor could be identified by analyzing published SNP profiles and associated genotypic and phenotypic information. We present the International Stem Cell Forum (ISCF) Ethics Working Party's Policy Statement on ' Publishing SNP Genotypes of Human Embryonic Stem Cell Lines (hESC)'. The Statement prospectively addresses issues surrounding the publication of genotypic data and associated annotations of hESC lines in open access databases. It proposes a balanced approach between the goals of open science and data sharing with the respect for fundamental bioethical principles (autonomy, privacy, beneficence, justice and research merit and integrity). [ABSTRACT FROM AUTHOR]

• lacZ codons optimized with 7 highly abundant cow milk proteins to construct olacZ. • olacZ expression increased in vitro, in cell lines 293T, HeLa, Cos7, NIH 3T3, HC11. • olacZ expression increased in vivo, in breast tissue in transgenic lactating female mice. • olacZ expression enhancement observed at the translational level. • System-preferential codon optimization can be used with other organs or bioreactors. lacZ encodes for β-galactosidase within the galactose operon of bacterial cells. When used as a reporter gene, bacterial "β-galactosidase" expression is often insufficient for detection in mammalian cells. We intended to optimize the lacZ codon usage according to the most frequently used codons for the seven major proteins in cow's milk, in order to pave a way for the enhancement of transgenic genes expression in eukaryotes. We constructed modified lacZ (named olacZ) according to optional codons used for proteins expressed in cow's milk. The expression of lacZ and olacZ was then compared in HC11 (a murine mammary gland epithelial line), 293T, HeLa, Cos7, and NIH 3T3 cells. While there was no significant difference at the mRNA level between lacZ and olacZ (P > 0.05). The quantification of β-galactosidase activity and in situ staining experiments showed a 1.2-fold to 3.3-fold expression improvement when comparing olacZ with lacZ. The levels of β-galactosidase expression at the protein levels from olacZ were approximately 9.2-fold and 2.4-fold respectively for Cos7 and HC11 cells. Furthermore, a 1.9-fold tendency of enhanced expression of olacZ in mammary gland during lactation was observed in transgenic- olacZ mice. This study demonstrates an alternative choice for improving lacZ reporter expression in eukaryotes, especially in the mammary gland of cattle or goats. [ABSTRACT FROM AUTHOR]

The fast charge carrier recombination and slow oxidation kinetics of ternary semiconductor InGaN as promising photocatalyst impede the PEC performance. Herein, we fabricate transition metal zinc (Zn) doped InGaN nanorods arrays by radio-frequency plasma-assisted molecular beam epitaxy. The doping obviously reduces indium atoms composition, the aggregation of In–In and induces the deep energy level. This greatly decreases the defects and improves the valence band potential of InGaN nanorods, which is beneficial for the rapid carrier separation efficiency with the decreased photogenerated carrier recombination rate and improved water oxidation kinetics. Significantly, Zn doped InGaN nanorods photoanode shows three times higher photocurrent density of 1.65 mA/cm2 at 1.23 V versus reversible hydrogen electrode (RHE) compared to undoped InGaN nanorods (0.58 mA/cm2). More importantly, after loading Au nanoparticles, the maximum applied bias photo-to-current efficiency of Zn-doped nanorods photoanode reaches 1.33%, which is superior to the recent results of reported Zn doped based photoanodes. This efficient doping strategy not only bridges the gaps of heteroatom doped InGaN nanorods based photoelectrodes, but also provides deep insights into controlling the electronic structure, and crystallinity of photoelectrodes for enhanced solar converting efficiency. Image 1 • Highly efficient Zn-doped InGaN NRs photoanodes are prepared. • Photogenerated carrier recombination is suppressed for Zn-doped InGaN NRs. • Zn doping contributes to the positive shift of valence band. • High photovoltage for Zn-doped InGaN NRs is achieved. [ABSTRACT FROM AUTHOR]

Objectives: This study explored whether TALENs‐mediated non‐homologous end joining (NHEJ) targeting the mutation site can correct the aberrant β‐globin RNA splicing, and ameliorate the β‐thalassaemia phenotype in β654 mice. Material and methods: TALENs vectors targeted to the human β‐globin gene (HBB) IVS2‐654C >T mutation in a mouse model were constructed and selected to generate double heterozygous TALENs+/β654 mice. The gene editing and off‐target effects were analysed by sequencing analysis. β‐globin expression was identified by RT‐PCR and Western blot analysis. Various clinical indices including haematologic parameters and tissue pathology were examined to determine the therapeutic effect in these TALENs+/β654 mice. Results: Sequencing analysis revealed that the HBBIVS2‐654C >T point mutation was deleted in over 50% of the TALENs+/β654 mice tested, and off‐target effects were not detected. RT‐PCR and Western blot analysis confirmed the expression of normal β‐globin in TALENs+/β654 mice. The haematologic parameters were significantly improved as compared with their affected littermates. The proportion of nucleated cells in bone marrow was considerably decreased, splenomegaly with extramedullary haematopoiesis was reduced, and significant decreases in iron deposition were seen in spleen and liver of the TALENs+/β654 mice. Conclusion: These results suggest effective treatment of the anaemia phenotype in TALENs+/β654 mice following deletion of the mutation site by TALENs, demonstrating a simple and straightforward strategy for gene therapy of β654‐thalassaemia in the future. [ABSTRACT FROM AUTHOR]