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Flowering time is an important agronomic trait for soybean yield and adaptation. However, the genetic basis of soybean adaptation to diverse latitudes is still not clear. Four NIGHT LIGHT-INDUCIBLE AND CLOCK-REGULATED 2 (LNK2) homeologs of Arabidopsis thaliana LNK2 were identified in soybean. Three single-guide RNAs were designed for editing the four LNK2 genes. A transgene-free homozygous quadruple mutant of the LNK2 genes was developed using the CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 (CRISPR-associated protein 9). Under long-day (LD) conditions, the quadruple mutant flowered significantly earlier than the wild-type (WT). Quantitative real-time PCR (qRT-PCR) revealed that transcript levels of LNK2 were significantly lower in the quadruple mutant than in the WT under LD conditions. LNK2 promoted the expression of the legume-specific E1 gene and repressed the expression of FT2a. Genetic markers were developed to identify LNK2 mutants for soybean breeding. These results indicate that CRISPR/Cas9-mediated targeted mutagenesis of four LNK2 genes shortens flowering time in soybean. Our findings identify novel components in flowering-time control in soybean and may be beneficial for further soybean breeding in high-latitude environments.

In this paper our group proposes and designs a lightweight convolutional neural network (CNN) for detecting facial emotions in real-time and in bulk to achieve a better classification effect. We verify whether our model is effective by creating a real-time vision system. This system employs multi-task cascaded convolutional networks (MTCNN) to complete face detection and transmit the obtained face coordinates to the facial emotions classification model we designed firstly. Then it accomplishes the task of emotion classification. Multi-task cascaded convolutional networks have a cascade detection feature, one of which can be used alone, thereby reducing the occupation of memory resources. Our expression classification model employs Global Average Pooling to replace the fully connected layer in the traditional deep convolution neural network model. Each channel of the feature map is associated with the corresponding category, eliminating the black box characteristics of the fully connected layer to a certain extent. At the same time, our model marries the residual modules and depth-wise separable convolutions, reducing large quantities of parameters and making the model more portable. Finally, our model is tested on the FER-2013 dataset. It only takes 3.1% of the 16GB memory, that is, only 0.496GB memory is needed to complete the task of classifying facial expressions. Not only can our model be stored in an 872.9 kilobytes file, but also its accuracy has reached 67% on the FER-2013 dataset. And it has good detection and recognition effects on those figures which are out of the dataset.

When TextRank algorithm based on graph model constructs graph associative edges, the co-occurrence window rules only consider the relationships between local terms. Using the information in the document itself is limited. In order to solve the above problems, an improved TextRank keyword extraction algorithm based on rough data reasoning combined with word vector clustering, RDD-WRank, was proposed. Firstly, the algorithm uses rough data reasoning to mine the association between candidate keywords, expands the search scope, and makes the results more comprehensive. Then, based on Wikipedia online open knowledge base, word embedding technology is used to integrate Word2Vec into the improved algorithm, and the word vector of TextRank lexical graph nodes is clustered to adjust the voting importance of nodes in the cluster. Compared with the traditional TextRank algorithm and the Word2Vec algorithm combined with TextRank, the experimental results show that the improved algorithm has significantly improved the extraction accuracy, which proves that the idea of using rough data reasoning can effectively improve the performance of the algorithm to extract keywords.

Hepatocyte necroptosis is a core pathogenetic event during alcoholic liver disease. This study was aimed to explore the potential of tetramethylpyrazine (TMP), an active hepatoprotective ingredient extracted from Ligusticum Wallichii Franch, in limiting alcohol-triggered hepatocyte necroptosis and further specify the molecular mechanism. Results revealed that TMP reduced activation of receptor-interacting protein kinase 1 (RIPK1)/RIPK3 necrosome in ethanol-exposed hepatocytes and phosphorylation of mixed-lineage kinase domain-like protein (MLKL), which thereby diminished necroptosis and leakage of damage-associated molecular patterns. Suppression on mitochondrial translocation of p-MLKL by TMP contributed to recovery of mitochondrial function in ethanol-damaged hepatocytes. TMP also disrupted necroptotic signal loop by interrupting mitochondrial reactive oxygen species (ROS)-dependent positive feedback between p-MLKL and RIPK1/RIPK3 necrosome. Further, TMP promoted clearance of impaired mitochondria in ethanol-incubated hepatocytes via restoring PINK1/parkin-mediated mitophagy. Ubiquinol-cytochrome c reductase core protein 2 (UQCRC2) was downregulated in ethanol-exposed hepatocytes, which was restored after TMP treatment. In vitro UQCRC2 knockdown lowered the capacities of TMP in reducing mitochondrial ROS accumulation, relieving mitochondria damage, and enhancing PINK1/parkin-mediated mitophagy in ethanol-exposed hepatocytes. Analogously, systematic UQCRC2 knockdown interrupted the actions of TMP to trigger autophagic signal, repress necroptotic signal, and protect against alcoholic liver injury, inflammation, and ROS overproduction. In conclusion, this work concluded that TMP rescued UQCRC2 expression in ethanol-challenged hepatocytes, which contributed to necroptosis inhibition by facilitating PINK1/parkin-mediated mitophagy. These findings uncovered a potential molecular pharmacological mechanism underlying the hepatoprotective action of TMP and suggested TMP as a promising therapeutic candidate for alcoholic liver disease.

Soybean (Glycine max (L.) Merrill) is an important legume crop worldwide. Plant height (PH) is a quantitative trait that is closely related to node number (NN) and internode length (IL) on the main stem, which together affect soybean yield. To identify candidate genes controlling these three traits in soybean, we examined a recombinant inbred line (RIL) population derived from a cross between two soybean varieties with semi-determinate stems (Dt1Dt1Dt2Dt2), JKK378 and HXW. A quantitative trait locus (QTL) named qPH18 was identified that simultaneously controls PH, NN, and IL; this region harbors the semi-determinant gene Dt2. Sequencing of the Dt2 promoter from JKK378 identified three polymorphisms relative to HXW, including two single nucleotide polymorphism (SNPs) and an 18-bp insertion/deletion polymorphism (Indel). Dt2 expression was lower in the qPH18(JKK378) group than in the qPH18(HXW) group, whereas the expression level of the downstream gene Dt1 showed the opposite tendency. A transient transfection assay confirmed that Dt2 promoter activity is lower in JKK378 compared to HXW. We propose that the polymorphisms in the dominant Dt2 promoter underlie the differences in Dt2 expression and its downstream gene Dt1 in the two parents, thereby affecting PH, NN, IL, and grain weight per plant without altering stem growth habit. Compared to the PH18(HXW) allele, the qPH18(JKK378) allele suppresses Dt2 expression, which releases the inhibition of Dt1 expression, thus enhancing NN and grain yield. Our findings shed light on the mechanism underlying NN and PH in soybean and provide a molecular marker to facilitate breeding. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s11032-021-01235-y.

The ZNF280D is a member of the C2H2 zinc finger family, which plays important roles in cell and organismal growth, secretion, and other biological processes. This study aimed to identify the alternative splicing of pig ZNF280D, and to predict the structure and function of the encoded proteins, as well as to study the spatio-temporal expression patterns of each transcript. RNA-seq and qPCR technology were used to identify the transcripts of pig ZNF280D. We found two transcripts, ZNF280D1 and ZNF280D2 (GenBank accession number: MF279143). ZNF280D1 is the common transcript, whereas ZNF280D2 lacked a large region at exon 8. In addition, ZNF280D2 had an additional C2H2 zinc finger domain, changing the protein from a tC2H2 to a maC2H2 zinc finger protein. The expression of two transcripts in various tissues of pig was differ significant. ZNF280D2 is the main subtype. The expression of ZNF280D2 in pig muscle was found to increase gradually with age. The ZNF280D1 was first decreased and then increased slightly. We successfully identified two transcripts of pig ZNF280D, and we suggested important functions for ZNF280D in digestive absorption and growth as well as in the development of skeletal muscles, the specific mechanisms for which would require further study.

Soybean [Glycine max (L.) Merrill] is one of the most important legume crops and its plant height (PH) is one significant quantitative trait closely related to node number (NN) and internode length (IL) on the main stem, which affect yield together. In this study, we used a recombinant inbred line (RIL) derived from a cross between two semi-determinate stem soybean varieties (Dt1Dt1Dt2Dt2), JKK378 and HXW. A consistent QTL named qPH18 simultaneously controlling PH, NN, and IL was identified, in which this region harbors the semi-determinant gene, Dt2. Sequencing of the promoter of Dt2 from JKK378, identified three polymorphisms relative to HXW, including two SNPs and a 18bp Indel. The expression level of Dt2 in qPH18JKK378 group was lower than that in qPH18HXW group, meanwhile, as a downstream gene, the expression of Dt1 from two groups showed a contrary tendency. Transient transfection assay confirmed that the promoter activity of Dt2 from JKK378 is lower compared to HXW. We speculate that the polymorphisms in the two dominant Dt2 promoter caused difference in expression level of Dt2 and its downstream gene Dt1, hence, affect PH, NN, IL, and grain weight per plant without changing stem growth habit. Compared to PH18HXW allele, qPH18JKK378 allele suppresses the expression of Dt2 gene which releases the expression of Dt1, thus promotes plant node number and enhanced grain yield. These results will be helpful for revealing the mechanism underlying the node number and plant height, the soybean materials and molecular marker will facilitate molecule breeding.

Asthma is a heterogeneous chronic inflammatory disorder of the airways with a complex etiology, which involves a variety of cells and cellular components. Therefore, the aim of the study was to investigate the effects and mechanisms of antagonistic peptides that specifically bind to the first and second extracellular loops of CCR5 (GH and HY peptides, respectively) and anti-interleukin-23 subunit p19 (anti-IL-23p19) in the airway and thereby mediate inflammation and the IL-23/T helper 17 (Th17) cell pathway in asthmatic mice. An experimental asthma model using BALB/c mice was induced by ovalbumin (OVA) and treated with peptides that are antagonistic to CCR5 or with anti-IL-23p19. The extents of the asthmatic inflammation and mucus production were assessed. In addition, bronchoalveolar lavage fluid (BALF) was collected, the cells were counted, and the IL-4 level was detected by ELISA. The IL-23/Th17 pathway-related protein and mRNA levels in the lung tissues were measured, and the positive production rates of Th17 cells in the thymus, spleen, and peripheral blood were detected. The groups treated with one of the two peptides and/or anti-IL-23p19 showed significant reductions in allergic inflammation and mucus secretion; decreased expression levels of IL-23p19, IL-23R, IL-17A and lactoferrin (LTF); and reduced proportions of Th17 cells in the thymus, spleen, and peripheral blood. Specifically, among the four treatment groups, the anti-IL-23p19 with HY peptide group exhibited the lowest positive production rate of Th17 cells. Our data also showed a significant and positive correlation between CCR5 and IL-23p19 protein expression. These findings suggest that the administration of peptides antagonistic to CCR5 and/or anti-IL-23p19 can reduce airway inflammation in asthmatic mice, most likely through inhibition of the IL-23/Th17 signaling pathway, and the HY peptide can alleviate inflammation not only through the IL-23/Th17 pathway but also through other mechanisms that result in the regulation of inflammation.

Purpose/Aim of the study: Cerebrovascular reactivity (CVR) is an important marker for assessing cerebrovascular disease. This study assessed the CVR by perfusion computed tomography (CT) and CO2 inhalation tests in patients with unilateral middle cerebral artery (MCA) stenosis disease. Materials and Methods: Thirty-one patients with unilateral MCA stenosis disease diagnosed by digital subtraction angiography were studied. Patients were divided into two groups according to the degree of stenosis: severe and moderate. The regional cerebral blood flow (CBF) before and after CO2 inhalation was determined by perfusion CT. Regional CVR values were obtained by the following formula: increase (%) = (post-CBF) − (pre-CBF)/(pre-CBF) × 100%. Results: No significant differences in the mean CBF in the MCA stenosis region were found between the affected and contralateral sides before the CO2 inhalation test; after the test, CBF was more significantly decreased on the affected side than on the contralateral side. The ch...

Purpose/aim of the study: Cerebrovascular reactivity (CVR) reflects the vasodilatory reserve of cerebral resistance vessels, which is an important marker for assessing cerebrovascular disease. The present study is to investigate whether CVR impairment increases adverse long-term outcome risk of patients with ≥ 50% symptomatic unilateral middle cerebral artery (MCA) stenosis (ischemic stroke (IS) or transient ischemic attack (TIA)).Digital subtraction angiography (DSA) was used to assess the degree of stenosis, and perfusion CT and 5% COSeventy-three patients with ≥ 50% symptomatic unilateral MCA stenosis, involving 31 non-CVR impairment cases and 42 CVR impairment cases, were included in the present study. Finally, IS occurred in six CVR impairment patients, and no endpoint happened in the non-CVR impairment group. Therefore, the annual rate of IS was 14.29% in the CVR impairment group and 0% in the non-CVR impairment group (P = 0.035). Besides, further Kaplan-Meier analysis found CVR impairment was closely associated with the IS risk (Kaplan-Meier Log-rank 4.719, P = 0.030).Our results showed that for patients with ≥ 50% symptomatic unilateral MCA stenosis, there was significant difference between non-CVR impairment cases and CVR impairment cases in the annual rate of IS. It suggests that CVR impairment increases the risk of adverse long-term outcomes.

Purpose/Aim of the study: Cerebrovascular reactivity (CVR) is an important marker for assessing cerebrovascular disease. This study assessed the CVR by perfusion computed tomography (CT) and COThirty-one patients with unilateral MCA stenosis disease diagnosed by digital subtraction angiography were studied. Patients were divided into two groups according to the degree of stenosis: severe and moderate. The regional cerebral blood flow (CBF) before and after CONo significant differences in the mean CBF in the MCA stenosis region were found between the affected and contralateral sides before the COCVR impairment was found in approximately half of patients with unilateral MCA stenosis. Along with an increase in the degree of stenosis, patients with unilateral MCA stenosis were more likely to exhibit CVR impairment. It is important to assess the CVR in patients with unilateral MCA stenosis, especially those with severe stenosis.