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Simple Summary: Porcine epidemic diarrhea virus (PEDV), porcine transmissible gastroenteritis virus (TGEV), porcine deltacoronavirus (PDCoV), and porcine rotavirus-A (PoRVA) are the four significant pathogens of viral diarrhea of piglets in large-scale pig farms, which have caused huge economic losses to the pig industry all over the world. Since these four viruses have very similar clinical symptoms, it is necessary to develop an excellent detection method that can differentiate and diagnose these four viruses. In this study, we developed a multiplex real-time RT-PCR method that can simultaneously differentiate and diagnose these four viruses. The method has high specificity and does not cross-react with other porcine viruses. The lower limit of detection was 2.18 × 102 copies/μL. In addition, we tested 97 clinical samples collected by this method, and the results were consistent with the detection results of traditional RT-PCR, indicating that this method is reliable. In summary, we developed a multiplex real-time RT-PCR method for simultaneous detection of PEDV, TGEV, PDCoV, and PoRVA, and the results of this study can provide technical means for the differential diagnosis and epidemiological investigation of these four porcine diarrhea virus diseases. Porcine epidemic diarrhea virus (PEDV), porcine transmissible gastroenteritis virus (TGEV), porcine deltacoronavirus (PDCoV), and porcine rotavirus-A (PoRVA) are the four main pathogens that cause viral diarrhea in pigs, and they often occur in mixed infections, which are difficult to distinguish only according to clinical symptoms. Here, we developed a multiplex TaqMan-probe-based real-time RT-PCR method for the simultaneous detection of PEDV, TGEV, PDCoV, and PoRVA for the first time. The specific primers and probes were designed for the M protein gene of PEDV, N protein gene of TGEV, N protein gene of PDCoV, and VP7 protein gene of PoRVA, and corresponding recombinant plasmids were constructed. The method showed extreme specificity, high sensitivity, and excellent repeatability; the limit of detection (LOD) can reach as low as 2.18 × 102 copies/μL in multiplex real-time RT-PCR assay. A total of 97 clinical samples were used to compare the results of the conventional reverse transcription PCR (RT-PCR) and this multiplex real-time RT-PCR for PEDV, TGEV, PDCoV, and PoRVA detection, and the results were 100% consistent. Subsequently, five randomly selected clinical samples that tested positive were sent for DNA sequencing verification, and the sequencing results showed consistency with the detection results of the conventional RT-PCR and our developed method in this study. In summary, this study developed a multiplex real-time RT-PCR method for simultaneous detection of PEDV, TGEV, PDCoV, and PoRVA, and the results of this study can provide technical means for the differential diagnosis and epidemiological investigation of these four porcine viral diarrheic diseases. [ABSTRACT FROM AUTHOR]

Objective: To explore the molecular etiology of Waardenburg syndrome type II (WS2) in a family from Yunnan province, China. Methods: A total of 406 genes related to hereditary hearing loss were sequenced using next‐generation sequencing. DNA samples were isolated from the peripheral blood DNA of probands. Those pathogenic mutations detected by next‐generation sequencing in probands and their parents were validated by Sanger sequencing. The conservatism of variation sites in genes was also analyzed. The protein expression was detected by flow cytometry. Results: A heterozygous mutation c.178delG (p.D60fs*49) in the SOX10 gene was identified in the proband, which is a frameshift mutation and may cause protein loss of function and considered to be a pathogenic mutation. This was determined to be a de novo mutation because her family were demonstrated to be wild‐type and symptom free. SOX10, FGFR3, SOX2, and PAX3 protein levels were reduced as determined by flow cytometry. Conclusion: A novel frameshift mutation in SOX10 gene was identified in this study, which may be the cause of WS2 in proband. In addition, FGFR3, SOX2, and PAX3 might also participate in promoting the progression of WS2. [ABSTRACT FROM AUTHOR]

As an important therapeutic target in breast cancer, HER2 expression assessed by immunohistochemistry plays a critical role in breast cancer treatment. Recent advances in HER2 antibody–drug conjugate therapy have enabled patients with HER2-low expression breast cancer to benefit from the drugs. However, it is not known whether the HER2-low expression in breast cancer FFPE blocks would be lost as storage time increased. In this study, we aimed to assess the loss of HER2 antigenicity in stored FFPE blocks of breast cancer and the rescue effect of modifying the protocol of antigen staining. We selected archived HER2-low breast cancer FFPE blocks with stored time ranging from 1 year to over 15 years and re-detected the expression of HER2. Our study showed that HER2 antigenicity loss increased with storage time and could cause false negativity in HER2-low detection. Moreover, we showed that by either increasing the antigen retrieval time or applying the tyramide signal amplification (TSA) kit, the HER2 signal can be rescued and detected in about half of the cases with HER2-low loss without causing false positivity. [ABSTRACT FROM AUTHOR]

Hepatoid adenocarcinoma of stomach (HAS) is a special subtype of gastric cancer with poor prognosis. Immunohistochemical analysis could provide important clues for the treatment of HAS. A total of 159 patients were diagnosed as HAS and 139 were enrolled in this study. Statistical differences were determined using relative test methods and survival analyses were performed by the Kaplan–Meier method to find survival differences. All tumors in this study were negative for Epstein–Barr virus‐encoded small RNAs (EBERs) and almost all showed no loss of mismatch repair (MMR) proteins and were positive for alpha fetoprotein (AFP or spalt like transcription factor 4 (SALL4). About half of the tumors had a positive programmed death‐ligand 1 combined positive score (CPS) and 17.3% were positive for human epidermal growth factor receptor 2 (HER2). In addition, there was a relatively high proportion of cmet expression. We also found that HAS patients with recurrent disease treated by emerging therapy had a better survival than those treated with traditional chemotherapy (p = 0.002, median recurrence‐to‐death survival: 23 months versus 6 months); HAS patients who received anti‐HER2 therapy or harbored MMR deficiency had favorable prognosis. Overall, high proportions of MMR protein proficiency, positivity for AFP or SALL4, overexpression of HER2, CPS and cmet, as well as negative EBER findings, are distinctive characteristics of HAS patients. While negative EBER and MMR proficiency indicate molecular features of HAS, positivity for AFP or SALL4 could aid in the diagnosis of HAS. In addition, HAS patients could benefit from anti‐HER2 therapy, immunotherapy, and anti‐angiogenesis therapy. [ABSTRACT FROM AUTHOR]

This paper reports the results of a review of research articles on the teaching of Chinese as a second or foreign language published in four leading mainland Chinese journals during the years 2005-2015. The review found that Chinese language researchers are exploring a wide array of issues including language policy and planning, language learning and use, language pedagogy, teacher development and language testing. These studies document the efforts that Chinese language researchers and teachers have made to meet the explosively growing demand for Chinese language learning in the world. Furthermore, it was noted that the leading Chinese journals have become more receptive to empirical studies although a large number of non-empirical articles are still being published. Overall, research in these journals is still beset with different challenges, and there is a need for more rigorous scholarship on the part of researchers. For this reason, the review concludes with suggestions for Chinese journals so that they can promote high-quality research to support the global development of Chinese language education.

This inquiry explored 68 pre-service perceptions and aspirations of Teaching Chinese as an International Language (TCIL) teachers by analysing the metaphors they used to describe themselves as teachers. The inquiry revealed that the participants used a variety of metaphors to project the images of themselves as pre-service TCIL teachers. The results also confirm that the participants' use of metaphors, as powerful tools to understand the complexity of teaching, reflects the mediation of cultural, historical and sociopolitical conditions. These findings help teacher educators to support TCIL teachers' professional development by engaging them in critical dialogues about the metaphors they use to perceive themselves as TCIL teachers. They also help teacher educators to adopt tailor-made pedagogical content to address their development needs.

Our previous studies have shown that the introduction of structurally diverse benzyl side chains at the C5-NH2 position of oseltamivir to occupy 150-cavity contributes to the binding affinity with neuraminidase and anti-influenza activity. To obtain broad-spectrum neuraminidase inhibitors, we designed and synthesised a series of novel oseltamivir derivatives bearing different N-heterocycles substituents that have been proved to induce opening of the 150-loop of group-2 neuraminidases. Among them, compound 6k bearing 4-((r)-2-methylpyrrolidin-1-yl) benzyl group exhibited antiviral activities similar to or weaker than those of oseltamivir carboxylate against H1N1, H3N2, H5N1, H5N6 and H5N1-H274Y mutant neuraminidases. More encouragingly, 6k displayed nearly 3-fold activity enhancement against H3N2 virus over oseltamivir carboxylate and 2-fold activity enhancement over zanamivir. Molecular docking studies provided insights into the explanation of its broad-spectrum potency against wild-type neuraminidases. Overall, as a promising lead compound, 6k deserves further optimisation by fully considering the ligand induced flexibility of the 150-loop. [ABSTRACT FROM AUTHOR]

To address drug resistance to influenza virus neuraminidase inhibitors (NAIs), a series of novel boron-containing N-substituted oseltamivir derivatives were designed and synthesized to target the 150-cavity of neuraminidase (NA). In NA inhibitory assays, it was found that most of the new compounds exhibited moderate inhibitory potency against the wild-type NAs. Among them, compound 2c bearing 4-(3-boronic acid benzyloxy)benzyl group displayed weaker or slightly improved activities against group-1 NAs (H1N1, H5N1, H5N8 and H5N1-H274Y) compared to that of oseltamivir carboxylate (OSC). Encouragingly, 2c showed 4.6 times greater activity than OSC toward H5N1-H274Y NA. Moreover, 2c exerted equivalent or more potent antiviral activities than OSC against H1N1, H5N1 and H5N8. Additionally, 2c demonstrated low cytotoxicity in vitro and no acute toxicity at the dose of 1000 mg/kg in mice. Molecular docking of 2c was employed to provide a possible explanation for the improved anti-H274Y NA activity, which may be due to the formation of key additional hydrogen bonds with surrounding amino acid residues, such as Arg152, Gln136 and Val149. Taken together, 2c appeared to be a promising lead compound for further optimization. [ABSTRACT FROM AUTHOR]

Plasminogen activator inhibitor-1 (PAI-1) is implicated in thrombogenesis, inflammation, and extracellular matrix remodeling. Previous studies indicated that oxidized low-density lipoprotein (LDL) stimulated the generation of PAI-1 in vascular endothelial cells (EC). The present study demonstrated that LDL oxidized by copper, iron, or 3-morpholinosydnonimine increased the expression of NADPH oxidase (NOX) 2, PAI-1, and heat shock factor-1 (HSF1) in human umbilical vein EC or coronary artery EC compared with LDL or vehicle. Diphenyleneiodonium, a NOX inhibitor, prevented the increases of the expression of HSF1 and PAI-1 in EC induced by oxidized LDLs. Small-interference RNA (siRNA) for [p22.sup.phox], an essential subunit of NOX, prevented oxidized LDL-induced expression of NOX2, HSF1, and PAI-1 in EC. HSF1 siRNA inhibited oxidized LDL-induced expression of PAI-1 and HSF1, but not NOX2, in EC. The binding of HSF1 to PAI-1 promoter and the activity of PAI-1 promoter in EC were enhanced by oxidized LDL. Butylated hydroxytulene, a potent antioxidant, inhibited oxidized LDL-induced release of hydrogen peroxide ([H.sub.2][O.sub.2]) and the expression of NOX2, HSF1, and PAI-1 in EC. Treatment with [H.sub.2][O.sub.2] increased the abundance of NOX2, HSF1, and PAI-1 in EC. The results of the present study indicate that oxidized LDL-induced expression of NOX may lead to the elevated release of reactive oxygen species, the activation of HSF1, and the enhancement of the transcription of PAI-1 gene in cultured vascular EC. reduced nicotinamide adenine dinucleotide phosphatase; oxidized low-density lipoprotein

Motivation Emerging single-cell RNA sequencing (scRNA-seq) technology empowers biological research at cellular level. One of the most crucial scRNA-seq data analyses is clustering single cells into subpopulations. However, the high variability, high sparsity and high dimensionality of scRNA-seq data pose lots of challenges for clustering analysis. Although many single-cell clustering methods have been recently developed, few of them fully exploit latent relationship among cells, thus leading to suboptimal clustering results. Results Here, we propose a novel unsupervised clustering method, scGAC (single-cell Graph Attentional Clustering), for scRNA-seq data. scGAC firstly constructs a cell graph and refines it by network denoising. Then, it learns clustering-friendly representation of cells through a graph attentional autoencoder, which propagates information across cells with different weights and captures latent relationship among cells. Finally, scGAC adopts a self-optimizing method to obtain the cell clusters. Experiments on 16 real scRNA-seq datasets show that scGAC achieves excellent performance and outperforms existing state-of-art single-cell clustering methods. Availability and implementation Python implementation of scGAC is available at Github (https://github.com/Joye9285/scGAC) and Figshare (https://figshare.com/articles/software/scGAC/19091348). Supplementary information Supplementary data are available at Bioinformatics online. [ABSTRACT FROM AUTHOR]

Aiming at the problems of low accuracy of English phrase part of speech recognition, poor English translation effect, and long translation time in the traditional English translation model, an English translation model based on intelligent recognition and deep learning is designed. An English phrase corpus was built, the phrase antecedent and postscript likelihood of the improved GLR algorithm by using the quaternion cluster were calculated, and the part of speech of the English phrase corpus was identified. According to the recognition results, the feature extraction algorithm is introduced to extract the best contextual features. On this basis, a neural machine translation model is constructed by integrating the traditional neural network in deep learning and combining the attention mechanism. It is used as a neural machine translation model for English translation. The simulation results show that the English translation model based on intelligent recognition and deep learning has high phrase recognition accuracy, good translation effect, and short translation time, which improves the quality of English translation. [ABSTRACT FROM AUTHOR]

This paper assesses the explanatory power of the liquidity-risk-based pricing models relative to the Fama–French three-factor model (FF3) and the extensions to the FF3. We find that the liquidity-augmented capital asset pricing model (LCAPM) performs no worse but generally better than other models considered in describing liquidity risk and a variety of anomaly portfolios. Our finding remains intact relative to the troublesome portfolios related to small, value, and aggressive investment.. This study highlights that liquidity risk is not negligible, which is in contrast to some recent findings that the price-impact-based liquidity risk factor contributes little to explain average returns. [ABSTRACT FROM AUTHOR]

Abstract Insulin-responsive glucose transporter type 4 (GLUT4) translocation plays a major role in controlling glucose uptake in adipose tissue and muscle, maintaining homeostasis and preventing hyperglycemia. Screening for chemicals enhancing GLUT4 translocation is an approach for identifying hits of drug development for type 2 diabetes. Here we developed a novel functional dual-color probe, pHluorin-GLUT4-mOrange2, and constructed 3T3-L1 adipocytes based screening system to simply and efficiently screen new compounds stimulating GLUT4 translocation. Based on this system, we successfully identified a few hits facilitating GLUT4 translocation. In conclusion, we developed an easy-to-apply dual color GLUT4 probe to monitor GLUT4 translocation in insulin-responsive cells, which could be alternatively employed to high-throughput screen compounds regulating GLUT4 translocation and glucose uptake, even to dissect GLTU4 approaching, docking and fusion with the plasma membrane (PM), and to reveal relevant molecular mechanisms involved in these steps as expected. Highlights • We develop an easy-to-apply dual color GLUT4 probe to monitor GLUT4 translocation. • PHluorin-GLUT4-mOrange2 is a useful tool for the study of signaling pathway involved in GLUT4 translocation. • PHluorin-GLUT4-mOrange2 can be applied to high throughput screening of GLUT4 transport activators easily and handily. [ABSTRACT FROM AUTHOR]

Outbreaks of duck Tembusu virus (DTMUV) have caused substantial economic losses in the major duck-producing regions of China since 2010. To improve our understanding of the host cellular responses to virus infection and the pathogenesis of DTMUV infection, we applied isobaric tags for relative and absolute quantification (iTRAQ) labeling coupled with multidimensional liquid chromatography-tandem mass spectrometry to detect the protein changes in duck embryo fibroblast cells (DEFs) infected and mock-infected with DTMUV. In total, 434 cellular proteins were differentially expressed, among which 116, 76, and 339 proteins were differentially expressed in the DTMUV-infected DEFs at 12, 24, and 42 hours postinfection, respectively. The Gene Ontology analysis indicated that the biological processes of the differentially expressed proteins were primarily related to cellular processes, metabolic processes, biological regulation, response to stimulus, and cellular organismal processes and that the molecular functions in which the differentially expressed proteins were mainly involved were binding and catalytic activity. Some selected proteins that were found to be differentially expressed in DTMUV-infected DEFs were further confirmed by real-time PCR. The results of this study provide valuable insight into DTMUV-host interactions. This could lead to a better understanding of DTMUV infection mechanisms. [ABSTRACT FROM AUTHOR]

Encrypted traffic classification (ETC) plays an important role in network management. In most research, the statistical features, transformed traffic images, or text are used for classification. However, the statistical features' design is time-consuming and labor-intensive, and the transformed traffic data lack spatial or semantic features. Considering that the headers of traffic packets have a uniform structure and are independent of each other, traffic data are most similar to tabular data. Thus we propose a data processing approach to convert packet headers into traffic tables in which each field is viewed as a column (feature). In addition, traffic data are hard to label in real traffic environments, and each field contributes differently to the classification. Therefore, a self-supervised learning algorithm, SubTab, is used as the baseline network to reduce the reliance on labeled data and assign different weights to different fields. To the best of our knowledge, this is the first time that the ETC problem is solved from the tabular domain. Experimental results on two real-world datasets, ISCX VPN-nonVPN and the self-collected dataset SHU-ET, demonstrate that our method surpasses state-of-the-art methods based on traffic images or text and proves that traffic tables are more suitable for ETC problems. In addition, our method achieves a great performance with only 10% of labeled data and reduces the reliance on labeling data. [ABSTRACT FROM AUTHOR]