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Infections with vancomycin-resistant Enterococcus (VRE) have been increasingly reported, especially in immunocompromised patients. Bacteriophages (phages) are often considered as potential therapeutic candidates in treating infectious diseases. To search for therapeutic phages as antimicrobial agents for the treatment of Enterococcus faecium infections, eight E. faecium phages were isolated from the environments. One of these, phage φEc-ZZ2, which was capable of eliciting efficient lysis of E. faecium, was selected for analysis. Transmission electron microscopy revealed its resemblance to members of the family Siphoviridae, with a tail 200 nm long and an icosahedral head 45 nm in diameter. Phage φEc-ZZ2 exhibited rapid adsorption (> 99% adsorbed in 10 min), a short latency period of 20 min and a large burst size of 180 PFU/cell. Host range determination shows that φEc-ZZ2 has a relatively broader host range (specificity for 54.5% of tested E. faecium strains). Furthermore, when φEc-ZZ2 alone was incubated at different pH, the phage was stable over a wide pH range (5 to 9). These characteristics of φEc-ZZ2 will provide useful information for further research on the interaction between Enterococcus phages and their hosts. Moreover, in consideration of its pH stability and broader host range, φEc-ZZ2 may be a good candidate as a therapeutic/disinfectant agent to control nosocomial infections caused by E. faecium.

Abstract The Intelligent Evaluation System for Calligraphy Characters (IESCC) is used for teaching calligraphy, and users can learn calligraphy through the modifications given by the system. Chinese character skeleton extraction is an important step in the intelligent evaluation algorithm of calligraphic characters. The skeletons of Chinese characters extracted by traditional refinement algorithms are prone to redundant branches and deformed skeletons, which can lead to skeleton extraction results that do not conform to the topology of the original character. In this study, the focus lies on hard‐pen regular script, and skeleton repair and extraction are performed for these characters. According to the writing characteristics of regular script, the redundant burs are removed and the deformation zone of the thinned skeleton is detected, and then the idea of first splitting is used, then restructuring, to propose a skeleton extraction algorithm based on stroke characterization and ambiguous zone detection for hard‐pen regular script, referred to as SCAD. First, a thinning algorithm is used to extract the skeleton of Chinese characters and remove redundant pixels. By analyzing the stroke characteristics of regular script, the burrs are classified and different conditions are set to detect and remove the burrs. Then the ambiguous zones are detected according to the different kinds of junction points. Then, curvature, stroke width and direction deviation are used to analyze the continuity of stroke segments, and the decision function is used to classify the stroke segments. Finally, the stroke segments with optimal pairings were compensated by interpolation according to the direction trend. This concludes the skeleton extraction. Skeleton extraction is performed on 1000 sample characters, and the SCAD algorithm can extract the skeleton of Chinese characters with an accuracy of up to 98.37%. It is proved that the SCAD method proposed here is a practical and effective method to extract the skeleton of hard‐pen regular script.

Abstract Background Dental caries in young children is a difficult global oral health problem. In the last decade, China has put a great deal of effort into reducing the prevalence of dental caries. This study, which is part of the China Population Chronic Disease and Nutrition Surveillance 2021, aimed to investigate the prevalence of dental caries among children aged 5 in Shanghai, China, and its associated factors. Methods A total of 1281 children aged 5 years from 6 districts in Shanghai were selected by a stratified sampling method. The survey consisted of an oral health questionnaire and an oral health examination. The questionnaire included questions on oral health knowledge, attitudes, and behaviours. The oral health examination used WHO standards. After screening, the data were input and analysed. Chi-square tests and logistic regression analyses were used to study the relevant factors affecting dental caries. Results The prevalence of dental caries among 1281 children was 51.0%, the dmft index score was 2.46, the Significant Caries Index (SiC) score was 6.39, and the SiC10 score was 10.35. Dental caries experience was related to the frequency of sweet drink consumption, the age of starting tooth brushing, eating habits after brushing, whether the children had received an oral examination provided by the government (p 0.05). Logistic regression analysis showed that the region of residence, eating after brushing and the age of starting brushing were associated with dental caries. Conclusions Dental caries remained prevalent among 5-year-old children in Shanghai, China. Prevention strategies that target the associated factors including region of residence, eating after brushing, and the age of starting brushing should be considered.

Abstract Swine acute diarrhea syndrome coronavirus (SADS-CoV), an emerging Alpha-coronavirus, brings huge economic loss in swine industry. Interferons (IFNs) participate in a frontline antiviral defense mechanism triggering the activation of numerous downstream antiviral genes. Here, we demonstrated that TRIM25 overexpression significantly inhibited SADS-CoV replication, whereas TRIM25 deficiency markedly increased viral yield. We found that SADS-CoV N protein suppressed interferon-beta (IFN-β) production induced by Sendai virus (SeV) or poly(I:C). Moreover, we determined that SADS-CoV N protein interacted with RIG-I N-terminal two caspase activation and recruitment domains (2CARDs) and TRIM25 coiled-coil dimerization (CCD) domain. The interaction of SADS-CoV N protein with RIG-I and TRIM25 caused TRIM25 multimerization inhibition, the RIG-I-TRIM25 interaction disruption, and consequent the IRF3 and TBK1 phosphorylation impediment. Overexpression of SADS-CoV N protein facilitated the replication of VSV-GFP by suppressing IFN-β production. Our results demonstrate that SADS-CoV N suppresses the host IFN response, thus highlighting the significant involvement of TRIM25 in regulating antiviral immune defenses.

Porcine epidemic diarrhea virus (PEDV) is associated with severe enteritis, which contributes to high mortality in piglets. The aim of this study was to describe molecular mechanisms associated with proinflammatory cytokine(s) production during PEDV infection. We showed that infection of porcine intestine epithelial cell clone J2 (IPEC-J2) with PEDV induces a gradual increase in interleukin 8 (IL-8) production at different time points, as well as infection of Vero E6 with PEDV. The secretion of IL-8 in these two cell lines infected with PEDV is related to the activation of NF-κB. Furthermore, the cells expressing PEDV M or E protein can induce the upregulation of IL-8. These findings suggest that the IL-8 production can be the initiator of inflammatory response by the host cells upon PEDV infection.

The cumulative coherence condition serves as a valuable tool within the realm of Compressed Sensing. However, its application to the $\ell _{1}-\ell _{2}$ -minimization model lacks thorough discussion. This paper aims to address this gap by introducing a sufficient condition for stable recovery of sparse vectors under the $\ell _{1}-\ell _{2}$ -minimization model, building upon the cumulative coherence condition. Moreover, employing graphical analysis, this study contrasts our proposed sufficient condition with existing criteria for stable recovery using both $\ell _{1}-\ell _{2}$ -minimization and $\ell _{1}$ -minimization models. Experimental data illustrate that our proposed sufficiency conditions exhibit less stringent requirements compared to established conclusions.

Abstract Ovarian cancer (OVCA), a prevalent gynecological malignancy, ranks as the fourth most common cancer among women. Mitotic Arrest Deficient 2 Like 2 (MAD2L2), a chromatin-binding protein and a component of DNA polymerase ζ, has been previously identified as an inhibitor of tumor growth in colorectal cancer. However, the roles of MAD2L2 in OVCA, including its expression, impact, and prognostic significance, remain unclear. We employed bioinformatics tools, Cox Regression analysis, and in vitro cell experiments to investigate its biological functions. Our findings reveal that MAD2L2 typically undergoes genomic alterations, such as amplifications and deep deletions. Moreover, we observed an overexpression of MAD2L2 mRNA in OVCA patients, correlating with reduced survival rates, particularly in those with Grade IV tumors. Furthermore, analysis of mRNA biofunctions indicated that MAD2L2 is predominantly localized in the organellar ribosome, engaging mainly in NADH dehydrogenase activity. This was deduced from the results of gene ontology enrichment analysis, which also identified its role as a structural constituent in mitochondrial translation elongation. These findings were corroborated by KEGG pathway analysis, further revealing MAD2L2’s involvement in tumor metabolism and the cell death process. Notably, MAD2L2 protein expression showed significant associations with various immune cells, including CD4+T cells, CD8+T cells, B cells, natural killer cells, and Myeloid dendritic cells. Additionally, elevated levels of MAD2L2 were found to enhance cell proliferation and migration in OVCA cells. The upregulation of MAD2L2 also appears to inhibit the ferroptosis process, coinciding with increased mTOR signaling activity in these cells. Our study identifies MAD2L2 as a novel regulator in ovarian tumor progression and offers new insights for treating OVCA.

In this study, a ring light point cloud calibration technique based on collimated laser beams is developed, aiming to reduce errors caused by the position and attitude changes of traditional ring light measurement devices. This article details the generation mechanism of the ring beam and the principle of deep hole measurement. It introduces the collimated beam as a reference, building on traditional ring light measurement devices, to achieve the synchronous acquisition of the ring beam and collimated spot images by an industrial camera. The Steger algorithm is employed to accurately extract the coordinates of the point cloud contours of both the ring beam and the collimated spot. By analyzing the shape and position changes of the collimated spot contour, the spatial position and attitude of the measuring device are precisely determined. This technique is applied to the 3D reconstruction of the inner surface of deep holes, ensuring the accurate restoration of the spatial positional attitude of the ring beam by incorporating the spatial positional attitude parameters of the measuring device to precisely calibrate the cross-sectional point cloud coordinates. Experimental results with ring gauges and deep hole workpieces demonstrate that this technique effectively reduces the percentage of point cloud data outside the tolerance range, and improves the accuracy of the 3D reconstruction model by 6.287%, thereby verifying the accuracy and practicality of this technique.

Adenosine kinase (ADK) is a key enzyme widely distributed in plants, playing an important role in maintaining cellular energy homeostasis and regulating plant growth, development, and responses to environmental stresses. However, research on ADK genes in cotton (Gossypium hirsutum), an economically significant crop, has been limited. This study identified 92 ADK genes from four cotton species (G. arboreum, G. raimondii, G. hirsutum, and G. barbadense) using HMMER and Local BLASTP methods and classified them into six groups. Chromosomal localization revealed a random distribution of ADK genes in G. hirsutum, with 13 genes located on the At subgenome and 14 genes on the Dt subgenome. Gene structure analysis showed consistency in exon–intron organization within subgroups, while conserved motif analysis identified subgroup-specific motifs, indicating functional diversity. Synteny and collinearity mapping analysis revealed that the primary expansion mechanisms of the ADK gene family in cotton are polyploidy and segmental duplication. Cis-regulatory elements in GhADK promoters were classified into light response, hormone response, developmental regulation, and stress response. We also analyzed the expression patterns of GhADK genes under a low temperature (4 °C) and drought conditions. Most GhADK genes responded to cold stress with different expression patterns, indicating their roles in rapid response and long-term cold adaptation. Under drought stress, expression patterns varied, with some genes showing sustained high expression levels. The qRT-PCR validation of transcriptomic data confirmed the stress-induced expression patterns of selected GhADK genes. Functional analysis through the VIGS silencing of GhADK25 demonstrated its importance in cold and drought stress responses, with silencing resulting in poor growth under stress, highlighting its significance in stress tolerance. This study provides a basis for further understanding the evolutionary relationships and functions of the cotton ADK gene family.

Porcine deltacoronavirus (PDCoV) is a novel enteric coronavirus that can cause vomiting, watery diarrhea in pigs and the death of piglets. The open reading frame (ORF) 5 is one of the accessory genes in PDCoV genome and encodes an accessory protein NS6. To date, the function of NS6 is still unclear. In this study, the recombinant NS6 was successfully expressed in prokaryotic expression system and purified. To prepare monoclonal antibody (mAb), six-week-old female BALB/c mice were primed subcutaneously with purified NS6. A novel mouse mAb against NS6 was obtained and designated as 3D5. The isotype of 3D5 is IgG2b with kappa (κ) light chain. 3D5 can specifically recognizes the natural NS6 in swine testis (ST) cells infected with PDCoV and expressed NS6 in human embryonic kidney 293T (HEK 293T) cells transfected with mammalian vector. The minimal linear B cell epitope recognised by 3D5 on NS6 was 25VPELIDPLVK34 determined by peptide scanning and named EP-3D5. The sequence of EP-3D5 is completely conserved among PDCoV strains. Moreover, six to nine residues of EP-3D5 were identified to be conserved in non-PDCoV strains. These results provide valuable insights into the antigenic structure and function of NS6 in virus pathogenesis, and aid for the development of PDCoV epitope-associated diagnostics and vaccine design.

In the original publication [...]

Objective: To evaluate the performance of newly developed glycated hemoglobin (HbA1c) clinical analytic reagents and HPLC columns, applied on Tosoh HLC-723 G8 Analyzer. Methods: Newly developed reagents and columns were used on a Tosoh HLC-723 G8 Analyzer (standard mode) system to measure both of qulity contorls and the clinical blood samples to evaluate the performances of these newly developed prodcuts including precision, accuracy, linearity, carryover, bias evaluation, correlation with commercial reagents, and stability according to CLSI recommendations. Results: The CV of intra-assay precision and inter-assay precision of quality control and clinical blood sample assays using Lirimax products were both less than 3.00%. And the REs of accuracy were less than 6.00%. Linearity: R2 = 0.9993 in the concentration range 4.77%–14.67%. Carryover: 0.05%. The Bland-Altman mean difference: −0.003583% HbA1c (CI: 0.07398: −0.08115); Passing-Bablok regression: y = 1.0022(0.9984:1.006)x-0.01097(-0.03776: 0.01582), R2 = 0.9996. Stability evaluation was also acceptable. Conclusion: The performance of newly developed products was well evaluated for HbA1c measurement on a TOSOH G8 Analyzer which shows excellent suitability for clinical assay.

Coronaviruses (CoVs) are a family of the largest RNA viruses that typically cause respiratory, enteric, and hepatic diseases in animals and humans, imposing great threats to the public safety and animal health. Porcine deltacoronavirus (PDCoV), a newly emerging enteropathogenic coronavirus, causes severe diarrhea in suckling piglets all over the world and poses potential risks of cross-species transmission. Here, we use PDCoV as a model of CoVs to illustrate the reciprocal regulation between CoVs infection and host antiviral responses. In this study, downregulation of DNA polymerase delta interacting protein 3 (POLDIP3) was confirmed in PDCoV infected IPEC-J2 cells by isobaric tags for relative and absolute quantification (iTRAQ) and Western blotting analysis. Overexpression of POLDIP3 inhibits PDCoV infection, whereas POLDIP3 knockout (POLDIP3-/-) by CRISPR-Cas9 editing significantly promotes PDCoV infection, indicating POLDIP3 as a novel antiviral regulator against PDCoV infection. Surprisingly, an antagonistic strategy was revealed that PDCoV encoded nonstructural protein 5 (nsp5) was responsible for POLDIP3 reduction via its 3C-like protease cleavage of POLDIP3 at the glutamine acid 176 (Q176), facilitating PDCoV infection due to the loss of antiviral effects of the cleaved fragments. Consistent with the obtained data in IPEC-J2 cell model in vitro, POLDIP3 reduction by cleavage was also corroborated in PDCoV infected-SPF piglets in vivo. Collectively, we unveiled a new antagonistic strategy evolved by PDCoV to counteract antiviral innate immunity by nsp5-mediated POLDIP3 cleavage, eventually ensuring productive virus replication. Importantly, we further demonstrated that nsp5s from PEDV and TGEV harbor the conserved function to cleave porcine POLDIP3 at the Q176 to despair POLDIP3-mediated antiviral effects. In addition, nsp5 from SARS-CoV-2 also cleaves human POLDIP3. Therefore, we speculate that coronaviruses employ similar POLDIP3 cleavage mechanisms mediated by nsp5 to antagonize the host antiviral responses to sustain efficient virus infection.

Highlights 1. The epidemiology of A. baumannii in Jilin Province showed distinctive local characteristic. 2. Newly discovered sequence types (STs), the results of genetic relatedness and geographical distribution of all STs showed epidemiological research by multi-locus sequence typing (MLST) in single hospital is helpful to monitor and control nosocomial infection of A. baumannii. 3. The rate of carbapenems resistance among A. baumannii is about 94% in Jilin province (northeast China). Carbapenems resistance is a helpful factor for predominant STs spreading.

In response to the challenges associated with the misassembly, omission, and low manual inspection efficiency in automobile wiring harness relay assemblies, a novel online detection system has been engineered. This system consists of a mobile-based visual imaging system and an improved YOLOv5-based detection algorithm that tracks human movement to acquire images and videos. The system is coupled with deep learning for real-time detection and recognition for error-proofing the installation process of automotive wiring harness relays. This innovation aims to facilitate error-proof inspection during the assembly process of automotive wiring harness relays. The YOLOv5s model is augmented with an Adaptive Spatial Feature Fusion (ASFF) module, enhancing multi-scale feature integration capabilities. A Global Context Network (GCNet) is incorporated into the C3 module to emphasize target information from a global perspective. Additionally, the replacement of standard Convolution (Conv) modules with Global Sparse Convolution (GSConv) modules in the Neck section effectively reduces computational costs while sustaining overall performance efficacy. The experimental results show that the detection system achieved a comprehensive accuracy rate of 99.2% and an F1 score of 99.29. The system possesses high accuracy and stability, enabling flexible and intelligent target detection applications in the automotive industry.

Swine acute diarrhea syndrome coronavirus (SADS-CoV) is an emerging porcine intestinal coronavirus that can cause acute diarrhea, vomiting, rapid weight loss, and high mortality in newborn piglets. Cholesterol 25-hydroxylase (CH25H) is a molecular mediator of innate antiviral immunity and converts cholesterol to 25-hydroxycholesterol (25HC). Previous studies have reported that CH25H and 25HC have an antiviral effect against multiple viruses. However, the interplay between SADS-CoV infection and CH25H or 25HC is still uncertain. Here, we found that CH25H and its enzymatic product 25HC restrained SADS-CoV replication by blocking membrane fusion. Our results show that CH25H was upregulated by SADS-CoV infection in vitro and in vivo, and that it was an IFN-stimulated gene in porcine ileum epithelial cells. Moreover, CH25H and CH25H mutants lacking catalytic activity can inhibit SADS-CoV replication. Furthermore, 25HC significantly suppressed SADS-CoV infection by inhibiting virus entry. Notably, we confirmed that CH25H and 25HC blocked SADS-CoV spike protein-mediated membrane fusion. Our data provide a possible antiviral therapy against SADS-CoV and other conceivable emerging coronaviruses in the future.

Autophagy plays an important role in the infectious processes of diverse pathogens. For instance, cellular autophagy could be harnessed by viruses to facilitate replication. However, it is still uncertain about the interplay of autophagy and swine acute diarrhea syndrome coronavirus (SADS-CoV) in cells. In this study, we reported that SADS-CoV infection could induce a complete autophagy process both in vitro and in vivo, and an inhibition of autophagy significantly decreased SADS-CoV production, thus suggesting that autophagy facilitated the replication of SADS-CoV. We found that ER stress and its downstream IRE1 pathway were indispensable in the processes of SADS-CoV-induced autophagy. We also demonstrated that IRE1-JNK-Beclin 1 signaling pathway, neither PERK-EIF2S1 nor ATF6 pathways, was essential during SADS-CoV-induced autophagy. Importantly, our work provided the first evidence that expression of SADS-CoV PLP2-TM protein induced autophagy through the IRE1-JNK-Beclin 1 signaling pathway. Furthermore, the interaction of viral PLP2-TMF451-L490 domain and substrate-binding domain of GRP78 was identified to activate the IRE1-JNK-Beclin 1 signaling pathway, and thus resulting in autophagy, and in turn, enhancing SADS-CoV replication. Collectively, these results not only showed that autophagy promoted SADS-CoV replication in cultured cells, but also revealed that the molecular mechanism underlying SADS-CoV-induced autophagy in cells.

IntroductionA study in 2006 showed that the clinical course of PEDV disease was markedly aggravated by transplacental infection of PCV2. Therefore, we investigated whether the small intestine supports PCV2 replication and the effect of PCV2 infection on PEDV replication in epithelial cells in vitro.MethodsTo confirm the intestinal tropism of PCV2, the viral loads in the small-intestinal tissues after PCV2 infection were determined with virus titration, and the viral titers in the infected pig jejunum, ileum, ileocecal valve, and colon were 104.86, 104.09, 102.52, and 102.35 TCID50/g, respectively. We then determined the propagation characteristics of PCV2 in ileal epithelial cells (IPI-FX) and jejunal epithelial cells (IPEC-J2) with an immunoperoxidase monolayer assay, virus titration, and an immunofluorescence assay. Both IPI-FX and IPEC-J2 cells supported the replication of PCV2, with titers of 105.5 and 105.0 TCID50/ml, respectively. We established an infection model of PCV2 and PEDV in IPI-FX cells and found that PEDV and PCV2 infected the cells individually and together. The effects of PCV2 infection on PEDV replication were determined with reverse transcription–quantitative PCR (qPCR), western blotting, and virus titration. When PCV2 infected IPI-FX cells before PEDV, PCV2 significantly inhibited the replication of PEDV in a dose- and time-dependent manner and that the mRNAs of IFN-β, TNF-α, IL1β, and OASL were downregulated (detected with qPCR). Surprisingly, when IPI-FX cells were co-infected with PCV2 and PEDV, PCV2 promoted the replication of PEDV, the expression of the host IFN-β, TNF-α, IL1β, and OASL mRNAs was upregulated.DiscussionThese findings demonstrate that the co-infection of IPI-FX cells with PCV2 and PEDV represents an excellent in vitro model in which to investigate their combined pathogenic mechanisms.

Multiprotein bridging factor 1 (MBF1) is an ancient family of transcription coactivators that play a crucial role in the response of plants to abiotic stress. In this study, we analyzed the genomic data of five Solanaceae plants and identified a total of 21 MBF1 genes. The expansion of MBF1a and MBF1b subfamilies was attributed to whole-genome duplication (WGD), and the expansion of the MBF1c subfamily occurred through transposed duplication (TRD). Collinearity analysis within Solanaceae species revealed collinearity between members of the MBF1a and MBF1b subfamilies, whereas the MBF1c subfamily showed relative independence. The gene expression of SlER24 was induced by sodium chloride (NaCl), polyethylene glycol (PEG), ABA (abscisic acid), and ethrel treatments, with the highest expression observed under NaCl treatment. The overexpression of SlER24 significantly enhanced the salt tolerance of tomato, and the functional deficiency of SlER24 decreased the tolerance of tomato to salt stress. SlER24 enhanced antioxidant enzyme activity to reduce the accumulation of reactive oxygen species (ROS) and alleviated plasma membrane damage under salt stress. SlER24 upregulated the expression levels of salt stress-related genes to enhance salt tolerance in tomato. In conclusion, this study provides basic information for the study of the MBF1 family of Solanaceae under abiotic stress, as well as a reference for the study of other plants.

Abstract Background The present study aimed to develop and test an extended theory of planned behavior (TPB), which includes attitudes, subjective norms and perceived behavioral control, oral health knowledge, and past oral health behavior on the intention to improve oral health behaviors among primary school students in Shanghai, China. Methods A school-based cross-sectional study was conducted with 414 students in the third-grade from 10 classes of Mingqiang Primary School located in Shanghai, China. Participants were recruited in October 2019. Data were collected through self-reported questionnaires, consisting of demographic characteristics, TPB variables, oral health knowledge and past oral health behaviors. Exploratory factor analysis was used to analyze TPB items. Pearson’s correlation and hierarchical regression analyses were conducted to identify the associated factors of intention to improve oral health behaviors. Results The study showed that among students in the third grade, attitudes, subjective norms, perceived behavioral control, and past oral health behaviors were associated with the intention to improve oral health behaviors. In the hierarchical regression analysis, age and sex were entered in Model 1 which significantly explained 3.00% of the variance (F = 6.26, p

Objective Achieving good oral health-related quality of life (OHRQOL) is of particular concern in children. The inter-relations among oral health knowledge, self-efficacy, behaviours and OHRQOL in children groups remain unclear. This study aimed to explore the inter-relations between these oral health behaviour-related factors and OHRQOL in primary school children.Methods In this cross-sectional study, 651 children in grades 2 and 3 were recruited in October 2020 from two primary schools in Minhang District, Shanghai, China. Data were collected through self-reported questionnaires, consisting of demographic characteristics, oral health knowledge, self-efficacy, oral health behaviours and OHRQOL. Pearson’s correlation analyses were used to analyse the relationship between study variables. Structural equation models were used to test the inter-relations between OHRQOL and oral health behaviour-related factors.Results Four hypothetical structural equation models were tested and one of them was selected as the most appropriate model, which explained 15.0% of the variance in OHRQOL. This selected model showed that oral health behaviours were directly related to OHRQOL. Oral health knowledge was indirectly associated with OHRQOL through both self-efficacy and oral health behaviours. Self-efficacy was directly associated with OHRQOL or was indirectly associated with OHRQOL through oral health behaviours.Conclusion This study revealed a pathway of association between children’s oral health knowledge and their OHRQOL, in which children’s oral health self-efficacy and behaviours had indirect effects. This provides a basis for understanding the mechanism of oral health promotion interventions to improve children’s OHRQOL and helps to identify direct or indirect intervention targets.

Swine acute diarrhea syndrome coronavirus (SADS-CoV) is an emerging swine enteric coronavirus that causes vomiting, severe diarrhea, dehydration and death in suckling piglets. NS7a is putative accessory protein that is predicted to be encoded by SADS-CoV, but still to be confirmed experimentally. In the present study, recombinant NS7a protein was expressed in a prokaryotic expression system and used as an antigen to prepare monoclonal antibodies (mAbs) specific to NS7a protein. We obtained two anti-NS7a mAbs, termed AH5 and EH3, that were shown by western blotting to react with the natural NS7a protein in Vero E6 cells infected with SADS-CoV. Using the produced mAbs, we observed by confocal microscopy that NS7a protein was expressed in the cytoplasm. Further studies revealed that the motif 31VNTWQEFA38 was the minimal unit of the linear B-cell epitope recognized by mAb AH5, and the motif 82FDLFERF88 was the minimal unit of the linear B-cell epitope recognized by mAb EH3. Alignment of amino acids showed that these two epitopes were highly conserved among different SADS-CoV strains and SADS-related coronaviruses from bats, but with one substitution in these two motifs in bat coronavirus HKU2. In summary, we generated and characterized two mAbs against SADS-CoV NS7a protein, and demonstrated NS7a expression in SADS-CoV-infected cells for the first time.

Abstract Purpose People with obesity have a compromised browning capacity of adipose tissue when faced with sympathetic stimuli. This study aimed to determine whether norepinephrine treatment can enhance the induction of precursor cells from human white adipose tissue to differentiate into adipocytes that express key markers of beige adipocytes, and if there is a difference in this capacity between normal weight and overweight individuals. Methods Stromal vascular cells derived from subcutaneous white adipose tissue of normal weight and overweight groups were induced to differentiation, with or without norepinephrine, into adipocytes. Oxygen consumption rate, lipolysis, the expression of uncoupling protein 1 and other thermogenic genes were compared between different adiposity and treatment groups. Results Peroxisome proliferator activated receptor γ- coactivator-1 alpha (PGC-1 α) and uncoupling protein 1 gene expression increased significantly in the normal weight group, but not in the overweight group, with norepinephrine treatment. The increments of lipolysis and oxygen consumption rate were also higher in adipocytes from the normal weight group with norepinephrine treatment, as compared with those of the overweight group. PR domain containing protein 16 (PRDM 16) gene expression was higher in the normal weight group compared with that in the overweight group, while there were no significant changes found with norepinephrine treatment in either the normal weight or overweight group. Conclusions Adipogenic precursor cells derived from overweight individuals were less prone to differentiate into beige-like adipocytes when facing sympathetic stimuli than normal weight ones, resulting in the compromised sympathetic-induced browning capacity in subcutaneous white adipose tissue in overweight individuals, which occurred before the onset of overt obesity.

ABSTRACTSwine acute diarrhea syndrome coronavirus (SADS-CoV), a newly discovered enteric coronavirus, is the aetiological agent that causes severe clinical diarrhea and intestinal pathological damage in piglets. To understand the effect of SADS-CoV on host cells, we characterized the apoptotic pathways and elucidated mechanisms underlying the process of apoptotic cell death after SADS-CoV infection. SADS-CoV-infected cells showed evidence of apoptosis in vitro and in vivo. The use of a pan-caspase inhibitor resulted in the inhibition of SADS-CoV-induced apoptosis and reduction in SADS-CoV replication, suggestive of the association of a caspase-dependent pathway. Furthermore, SADS-CoV infection activated the initiators caspase-8 and -9 and upregulated FasL and Bid cleavage, demonstrating a crosstalk between the extrinsic and intrinsic pathways. However, the proapoptotic proteins Bax and Cytochrome c (Cyt c) relocalized to the mitochondria and cytoplasm, respectively, after infection by SADS-CoV. Moreover, Vero E6 and IPI-2I cells treated with cyclosporin A (CsA), an inhibitor of mitochondrial permeability transition pore (MPTP) opening, were completely protected from SADS-CoV-induced apoptosis and viral replication, suggesting the involvement of cyclophilin D (CypD) in these processes. Altogether, our results indicate that caspase-dependent FasL (extrinsic)- and mitochondria (intrinsic)- mediated apoptotic pathways play a central role in SADS-CoV-induced apoptosis that facilitates viral replication. In summary, these findings demonstrate mechanisms by which SADS-CoV induces apoptosis and improve our understanding of SADS-CoV pathogenesis.

At present, the use of efficient and cost-effective methods to construct plasmonic surface-enhanced Raman scattering (SERS) substrates of high sensitivity, uniformity and reproducibility is still crucial to satisfy the practical application of SERS technology. In this paper, a localized surface plasmonic resonance (LSPR) tunable flexible Ag@PDMS substrate was successfully constructed by the low-cost bio-template-stripping method and magnetron sputtering technology. The theory proves that the local electromagnetic field enhancement and “hot spot” distribution is adjustable by modifying the size of the optical cavity unit in the periodicity nanocavity array structure. Experimentally, using rhodamine 6G (R6G) as the target analyte, the SERS performance of optimal Ag@PDMS substrate (Ag film thickness for 315 nm) was researched in detail, which the minimum detection limit was 10−11 M and the enhancement factor was calculated as 8.03 × 108, indicating its high sensitivity. The relative standard deviation (RSD) was calculated as 10.38%, showing that the prepared substrate had excellent electromagnetic field enhancement uniformity. At last, the trace detection of Crystal violet (CV, LOD = 10−9 M) and the simultaneous detection of three common dyes (R6G, CV and Methylene blue (MB) mixture) were also realized. This result suggests that the SERS substrate has a good application prospect in the quantitative and qualitative detection of dye molecules.

Swine enteric disease is the predominant cause of morbidity and mortality, and viral species involved in swine enteric disease include rotaviruses and coronaviruses, among others. Awareness of the circulating porcine rotavirus group C (PoRVC) in pig herds is critical to evaluate the potential impact of infection. At present, due to the lack of disease awareness and molecular diagnostic means, the research on RVC infection in China is not well-studied. In this study, diarrhea samples collected from pig farms were detected positive for RVC by PCR, and the full-length RVC was not previously reported for Chinese pig farms. This rotavirus strain was designated as RVC/Pig/CHN/JS02/2018/G6P6. A natural recombination event was observed with breakpoints at nucleotides (nt) 2509 to 2748 of the VP2 gene. Phylogenetic analysis based on nsp1 revealed that a new branch A10 formed. Collectively, our data suggest a potentially novel gene recombination event of RVC in the VP2 gene. These findings provide a new insight into the evolution of the rotavirus.

BackgroundThyroid nodules are a very common finding in the general population. Fine-needle aspiration (FNA) has been recommended as the initial test for the evaluation of thyroid nodules. The trend on reporting as atypia of undetermined significance (AUS) has been significantly increased, but the malignant risk is still controversial among different studies. The aim of this study is to investigate the malignancy risk of thyroid nodules reported as Bethesda category III (AUS/FLUS) on initial FNA.MethodWe reviewed 272 papillary thyroid cancer (PTC) patients with suspicious thyroid nodules who underwent fine-needle aspiration and received surgical treatment during 2019 to 2020.ResultsOne hundred ten (40.4%) patients were diagnosed with PTC. Multivariate analysis showed that microcalcification (p = 0.037, OR = 2.260, 95% CI: 1.051–4.860), shape (p = 0.003, OR = 4.367, 95% CI: 1.629–11.705), diameters (p = 0.002, OR = 0.278, 95% CI: 0.123–0.631), anti-thyroglobulin antibodies (TGAb) (p = 0.002, OR = 0.150, 95% CI: 0.046–0.494), anti-thyroid peroxidase antibody (A-TPO) (p = 0.009, OR = 4.784, 95% CI: 1.486–15.401), and nodule goiter (p < 0.001, OR = 0.100, 95% CI: 0.046–0.217) were independent malignant risk factors in patients with thyroid nodule classified as Bethesda category III.ConclusionIn this study, malignant risk factors in patients with thyroid nodule classified as Bethesda category III were significantly associated with preoperative serum TGAb, A-TPO, microcalcification, irregular shape, and nodule diameters. Nodules with malignant factors should be carefully elevated; surgery may be the better option for those patients.

In this work, a rapid and controllable ultrasonic vibration method for forming Al-based metallic glass at room temperature is proposed. This method can dramatically improve the forming ability of Al-based metallic glasses, which are virtually brittle at room temperature and have almost no supercooled liquid region at high temperatures. Under ultrasonic vibration, Al-based metallic glasses exhibited obvious plastic flow, with a maximum deformation degree up to 58% and an average deformation degree up to 43%. It is worth mentioning that no crystalline peaks were found on the X-ray diffraction patterns after deformation under ultrasonic vibration, and the mechanical properties remained the same as the primary sample. The present results provide a new approach for the deformation and forming of Al-based metallic glasses, which can significantly broaden their applications.

A hybrid process combining ultrasonic injection molding and electrical discharge machining was proposed to realize the economic, high-precision, and environmental-friendliness production of glass fiber reinforced polypropylene microstructured parts. First, the prism array microstructures were processed on tungsten carbide-cobalt (WC-8 wt% Co) substrate by low-speed wire electrical discharge machining and used as the mold core. Then, the glass fiber reinforced polypropylene composite (GF/PP) was melted under ultrasonic vibration, and the surface microstructure of the mold core was replicated. The experimental results show that this method can efficiently and accurately manufacture microstructured parts, and the microstructure has a high replication rate up to 95.5%. By replicating the microstructure, the surface contact angle of GF/PP parts increased from 57.1° to 134.9°, and the wettability changed from hydrophilicity to hydrophobicity. The mechanical properties test results show that the GF/PP parts fabricated by ultrasonic injection molding had excellent mechanical properties, and the 30% GF/PP parts show the highest tensile strength of 56.9 MPa. This work provides a new option for fabricating fiber reinforced composite parts.

Two important basic inference models of fuzzy reasoning are Fuzzy Modus Ponens (FMP) and Fuzzy Modus Tollens (FMT). In order to solve FMP and FMT problems, the full implication triple I algorithm, the reverse triple I algorithm and the Subsethood Inference Subsethood (SIS for short) algorithm are proposed, respectively. Furthermore, the existing reasoning algorithms are extended to intuitionistic fuzzy sets and interval-valued fuzzy sets according to different needs. The purpose of this paper is to study the relationship between intuitionistic fuzzy reasoning algorithms and interval-valued fuzzy reasoning algorithms. It is proven that there is a bijection between the solutions of intuitionistic fuzzy triple I algorithm and the interval-valued fuzzy triple I algorithm. Then, there is a bijection between the solutions of intuitionistic fuzzy reverse triple I algorithm and the interval-valued fuzzy reverse triple I algorithm. At the same time, it is shown that there is also a bijection between the solutions of intuitionistic fuzzy SIS algorithm and interval-valued fuzzy SIS algorithm.

Porcine deltacoronavirus (PDCoV) is a recently discovered enteropathogenic coronavirus and has caused significant economic impacts on the pork industry. Although studies have partly uncovered the molecular mechanism of PDCoV–host interaction, it requires further research. In this study, we explored the roles of Stromal Antigen 2 (STAG2) in PDCoV infection. We found that STAG2-deficient cells inhibited infection with vesicular stomatitis virus (VSV) and PDCoV, whereas restoration of STAG2 expression in STAG2-depleted (STAG2−/−) IPEC-J2 cells line restored PDCoV infection, suggesting that STAG2 is involved in the PDCoV replication. Furthermore, we found that STAG2 deficiency results in robust interferon (IFN) expression. Subsequently, we found that STAG2 deficiency results in the activation of JAK-STAT signaling and the expression of IFN stimulated gene (ISG), which establish an antiviral state. Taken together, the depletion of STAG2 activates the JAK-STAT signaling and induces the expression of ISG, thereby inhibiting PDCoV replication. Our study provides new insights and potential therapeutic targets for unraveling the mechanism of PDCoV replication.

Porcine epidemic diarrhea virus (PEDV) infects piglets and causes serious diarrhea as well as vomiting, dehydration, and death. The trimeric S protein plays a crucial role in the induction of neutralizing antibodies, and many neutralizing monoclonal antibodies (mAbs) against PEDV S protein have been developed. However, these mAbs exclusively target the S1 protein. In this study, we obtained a neutralizing mAb, 5F7, against the S2 protein of PEDV, and this mAb could neutralize new variant genotype 2 PEDV strains (LNCT2), as well as a genotype 1 PEDV strain (CV777), in vitro. The core sequence of the epitope was found in amino acid sequence 1261 aa~1337 aa. These findings confirm that the S2 protein possessed neutralizing epitopes and provided knowledge to aid further research on this virus.

The B-box proteins (BBXs) are a family of zinc-finger transcription factors with one/two B-Box domain(s) and play important roles in plant growth and development as well as stress responses. Wolfberry (Lycium barbarum L.) is an important traditional medicinal and food supplement in China, and its genome has recently been released. However, comprehensive studies of BBX genes in Lycium species are lacking. In this study, 28 LbaBBX genes were identified and classified into five clades by a phylogeny analysis with BBX proteins from Arabidopsis thaliana and the LbaBBXs have similar protein motifs and gene structures. Promoter cis-regulatory element prediction revealed that LbaBBXs might be highly responsive to light, phytohormone, and stress conditions. A synteny analysis indicated that 23, 20, 8, and 5 LbaBBX genes were orthologous to Solanum lycopersicum, Solanum melongena, Capsicum annuum, and Arabidopsis thaliana, respectively. The gene pairs encoding LbaBBX proteins evolved under strong purifying selection. In addition, the carotenoid content and expression patterns of selected LbaBBX genes were analyzed. LbaBBX2 and LbaBBX4 might play key roles in the regulation of zeaxanthin and antheraxanthin biosynthesis. Overall, this study improves our understanding of LbaBBX gene family characteristics and identifies genes involved in the regulation of carotenoid biosynthesis in wolfberry.

Large‐volume central lymph node metastasis (large-volume CLNM) is associated with high recurrence rate in papillary thyroid carcinoma (PTC) patients. However, sensitivity in investigating large-volume CLNM on preoperative ultrasonography (US) is not high. The aim of this study is to investigate the clinical factors associated with large-volume CLNM in clinical N0 PTC patients. We reviewed 976 PTC patients undergoing total thyroidectomy with central lymph node dissection during 2017 to 2019. The rate of large-volume LNM was 4.1% (40 of 967 patients). Multivariate analysis showed that male gender and young age (age2cm) was associated with large-volume CLNM, with OR (95% CI) 3.757 (1.061–13.310), by multivariate analysis. In ROC curve analysis on the diameter of the CPTC tumor, the Area Under Curve (AUC) =0.682(p=0.013), the best cut-off point was selected as 2.0cm. In conclusion, male gender and young age were predictors for large-volume CLNM of cN0 PTC. cN0 PTMC patient with capsule invasion and cN0 CPTC patient with tumor diameter >2cm were correlated with large-volume CLNM. Total thyroidectomy with central lymph node dissection may be a favorable primary treatment option for those patients.

Abstract Mitochondrial DNA depletion syndrome (MDS) is a group of severe inherited disorders caused by mutations in genes, such as deoxyribonucleoside kinase (DGUOK). A great majority of DGUOK mutant MDS patients develop iron overload progressing to severe liver failure. However, the pathological mechanisms connecting iron overload and hepatic damage remains uncovered. Here, two patients’ skin fibroblasts are reprogrammed to induced pluripotent stem cells (iPSCs) and then corrected by CRISPR/Cas9. Patient‐specific iPSCs and corrected iPSCs‐derived high purity hepatocyte organoids (iHep‐Orgs) and hepatocyte‐like cells (iHep) are generated as cellular models for studying hepatic pathology. DGUOK mutant iHep and iHep‐Orgs, but not control and corrected one, are more sensitive to iron overload‐induced ferroptosis, which can be rescued by N‐Acetylcysteine (NAC). Mechanically, this ferroptosis is a process mediated by nuclear receptor co‐activator 4 (NCOA4)‐dependent degradation of ferritin in lysosome and cellular labile iron release. This study reveals the underlying pathological mechanisms and the viable therapeutic strategies of this syndrome, and is the first pure iHep‐Orgs model in hereditary liver diseases.

In the current experimental study, we scrutinized the chemoprotective effect of astraxanthin against the 7,12-dimethylbenz(a)anthracene (DMBA)-induced breast cancer via Nrf-2-Keap1 and NF-kB and mTOR/Maf-1/PTEN pathway. The double emulsion solvent displacement method was used for the preparation of astraxanthin solid lipid nanoparticles (SLN). SLNs were appraised for entrapment, potential, size, drug-release performance, and gastric stability. DMBA (8 mg/kg) was used for the induction of breast cancer. Tumor weight, body weight, and tumor incidence were estimated at a regular interval. Different biochemical parameters such as Na+/K+, Ca2+, and Mg2+ activity, antioxidant, lipid, glycoprotein, phase I and II biotransformation enzymes, mitochondrial TCA cycle, and carbohydrate metabolizing enzymes were estimated. Keap1-Nrf-2, associated HO-1, and NF-kB expressions were estimated. Moreover, it estimated the mRNA expression of LXR (α,β), HMG-CoAR, PTEN, Maf1, PI3K, mTOR, Akt, FASN, and ACC1. AX-SLN reduced the tumor incidence, tumor weight, and increased the body weight. AX-SLN exhibited the protective effect against the LPO, enzymic (SOD, CuZnSOD, MnSOD, GPx, and CAT), and nonenzymic (GSH) in the serum, mammary gland, renal, and hepatic tissues. AX-SLN reduced the p-AKT which is accountable for the reduction in the NF-kB expression and also reduced the expression of Keap1 and NF-kB along with increasing the expression of HO-1 and Nrf-2. Further, AX-SLN significantly altered the mRNA of LXR (α,β), HMG-CoAR, PTEN, Maf1, PI3K, mTOR, Akt, FASN, and ACC1. On the basis of the results, we can conclude that AX-SLN inhibits the mammary gland carcinogenesis via Nrf-2-Keap1, NF-kB, and mTOR/Maf-1/PTEN pathway.

Objective To investigate the characteristics of diabetic foot ulcer (DFU) recurrence. Methods A total of 573 patients with DFUs were recruited and divided into an initial group (395 patients) and a recurrence group (178 patients). The factors related to recurrence were analyzed using multivariate regression. Results The recurrence group had longer diabetes duration (odds ratio [OR] 192; 95% confidence interval 120, 252 vs. 156; 96, 240); lower glycated hemoglobin levels (OR 8.1; 95% CI 6.8, 9.6 vs. 9.1; 7.4, 10.5), and higher rates than the initial group of amputation (37.5% vs. 2.0%), history of vascular intervention (21.3% vs. 3.9%), retinopathy (77.7% vs. 64.7%), callus (44.4% vs. 20.8%), foot deformity (51.2% vs. 24.6%), and outdoor sports shoe wearing (34.0% vs. 21.2%). Multiple factor logistic regression analysis showed that diabetes duration (OR 1.004), callus (OR 2.769), vascular intervention (OR 2.824) and amputation (OR 22.256) were independent risk factors for DFU recurrence. Conclusion Diabetes duration, callus, history of vascular intervention, and amputation were independent risk factors for recurrent DFUs in a cohort of Chinese patients with active DFU. The prevention and treatment of DFUs, especially callus treatment, foot care, and blood glucose control, should be improved in China.

The R2R3-MYB is a large gene family involved in various plant functions, including carotenoid biosynthesis. However, this gene family lacks a comprehensive analysis in wolfberry (Lycium barbarum L.) and other Solanaceae species. The recent sequencing of the wolfberry genome provides an opportunity for investigating the organization and evolutionary characteristics of R2R3-MYB genes in wolfberry and other Solanaceae species. A total of 610 R2R3-MYB genes were identified in five Solanaceae species, including 137 in wolfberry. The LbaR2R3-MYB genes were grouped into 31 subgroups based on phylogenetic analysis, conserved gene structures, and motif composition. Five groups only of Solanaceae R2R3-MYB genes were functionally divergent during evolution. Dispersed and whole duplication events are critical for expanding the R2R3-MYB gene family. There were 287 orthologous gene pairs between wolfberry and the other four selected Solanaceae species. RNA-seq analysis identified the expression level of LbaR2R3-MYB differential gene expression (DEGs) and carotenoid biosynthesis genes (CBGs) in fruit development stages. The highly expressed LbaR2R3-MYB genes are co-expressed with CBGs during fruit development. A quantitative Real-Time (qRT)-PCR verified seven selected candidate genes. Thus, Lba11g0183 and Lba02g01219 are candidate genes regulating carotenoid biosynthesis in wolfberry. This study elucidates the evolution and function of R2R3-MYB genes in wolfberry and the four Solanaceae species.

In symmetric environment, as the tangential electric field is almost constant across the ultrathin film, the absorption rate of the film is up to 0.5 under single side illumination. In this letter, we propose a symmetrical single layer ultrathin metal grating structure where perfect absorption (up to 0.99) can be achieved with unilateral illumination. We consider that this phenomenon of extraordinary optical absorption is caused by the coupling of the localized surface plasmon resonance and the long range surface plasmon. In particular, the electromagnetic response of the metal grating we proposed greatly resembles the electromagnetic induced absorption (EIA) which shows a narrow absorption peak within a broad absorption profile.

Enterobacter cloacae, an important agent associated with nosocomial infection, usually involves expanded-spectrum cephalosporin resistance. The therapeutic potential of bacteriophage is a possibly alternative chemotherapy against bacterial infection. In this study, we have characterized one newly isolated bacteriophage φEC14, which is lytic to E. cloacae specifically. Transmission electron microscopy revealed that phage φEC14 had an icosahedral head and long contractile tail, morphologically similar to phages belonging to family Siphoviridae. Pulsed-Field Gel Electrophoresis (PFGE) showed that the size of φEC14 virion DNA was in rang of 23.0-48.5 kb. Restriction analysis showed that lytic phage φEC14 was a double-stranded DNA virus, which might be cut by some restriction endonucleases. SDS-PAGE of phage proteins exhibited one major band and six minor bands with molecular weight ranging from 6.5 to 66.4 kilo-Dalton. In one-step experiment, phage φC14 had a short latent period of 10 minutes and a burst size of 50 PFU/cell. The best understanding of the biological features of lytic bacteriophage φEC14 would facilitate the development of an alternative agent to control the spread of multidrug-resistant E. cloacae.

Abstract Self-assembled organic-inorganic CH3NH3PbI3 perovskite microwires (MWs) upon humidity exposure along several weeks were investigated by photoluminescence (PL) spectroscopy, Raman spectroscopy, and X-ray diffraction (XRD). We show that, in addition to the common perovskite decomposition into PbI2 and the formation of a hydrated phase, humidity induced a gradual PL redshift at the initial weeks that is stabilized for longer exposure (~ 21 nm over the degradation process) and an intensity enhancement. Original perovskite Raman band and XRD reflections slightly shifted upon humidity, indicating defects formation and structure distortion of the MWs crystal lattice. By correlating the PL, Raman, and XRD results, it is believed that the redshift of the MWs PL emission was originated from the structural disorder caused by the incorporation of H2O molecules in the crystal lattice and radiative recombination through moisture-induced subgap trap states. Our study provides insights into the optical and structural response of organic-inorganic perovskite materials upon humidity exposure.

Knowing the droplet-deformation conditions, the droplet-breakup conditions, and the drag force in the interaction between two droplets with a high Reynolds number is of importance for tracking droplet movement in the annular flow field of a gas well. The interactions between two droplets with a high Reynolds number in a tandem arrangement fixed in flowing gas was investigated. The volume of fluid (VOF) method was used to model the droplets’ surface structure. Two different body forces were exerted on both droplets to hold them suspended at a fixed location, which eliminated the effect of droplet acceleration or deceleration on the drag and decreased the amount of computation required. The exerted body forces were calculated using the Newton iteration procedure. The interactions between the two droplets were analyzed by comparison with the simulation results of a single isolated droplet. The effect of the separation distance on the drag force was investigated by changing the separation spacing. The simulation results showed that for droplets with a small separating space between them, the dynamics of the downstream droplet were influenced significantly by the upstream droplet. The drag coefficient of the downstream droplet decreased considerably to a small, even negative, value, especially for droplets with higher Weber numbers and smaller initial separating spaces between them, while the drag force of the upstream droplet was influenced only slightly. In addition, a formula for predicting the final drag coefficient of the downstream droplet was devised.

The emissions of greenhouse gases (GHGs) from inland waters are an important component of the global carbon (C) cycle. However, the current understanding of GHGs emissions from arid river systems remains largely unknown. To shed light on GHGs emissions from inland waters in arid regions, high-resolution carbon dioxide (CO2) and methane (CH4) emission measurements were carried out in the arid Kuye River Basin (KRB) on the Chinese Loess Plateau to examine their spatio-temporal variability. Our results show that all streams and rivers were net C sources, but some of the reservoirs in the KRB became carbon sinks at certain times. The CO2 flux (FCO2) recorded in the rivers (91.0 mmol m−2 d−1) was higher than that of the reservoirs (10.0 mmol m−2 d−1), while CH4 flux (FCH4) in rivers (0.35 mmol m−2 d−1) was lower than that of the reservoirs (0.78 mmol m−2 d−1). The best model developed from a number of environmental parameters was able to explain almost 40% of the variability in partial pressure of CO2 (pCO2) for rivers and reservoirs, respectively. For CH4 emissions, at least 70% of the flux occurred in the form of ebullition. The emissions of CH4 in summer were more than threefold higher than in spring and autumn, with water temperature being the key environmental variable affecting emission rates. Since the construction of reservoirs can alter the morphology of existing fluvial systems and consequently the characteristics of CO2 and CH4 emissions, we conclude that future sampling efforts conducted at the basin scale need to cover both rivers and reservoirs concurrently.

Transmissible gastroenteritis virus (TGEV) is an infective coronavirus (CoV) that causes diarrhea-related morbidity and mortality in piglets. For the first time, a natural recombination strain of a TGEV Anhui Hefei (AHHF) virus between the Purdue and the Miller clusters was isolated from the small intestine content of piglets in China. A phylogenetic tree based on a complete genome sequence placed the TGEV AHHF strain between the Purdue and the Miller clusters. The results of a computational analysis of recombination showed that the TGEV AHHF strain is a natural recombinant strain between these clusters. Two breakpoints located in the open reading frame 1a (ORF1a) and spike (S) genes were identified. The pathogenicity of the TGEV AHHF strain was evaluated in piglets, and the results show that TGEV AHHF is an enteric pathogenic strain. These results provide valuable information about the recombination and evolution of CoVs and will facilitate future investigations of the molecular pathogenesis of TGEV.Emerging Microbes & Infections (2017) 6, e74; doi:10.1038/emi.2017.62; published online 23 August 2017

The hydraulic lifting system is the key part of the crane. A sliding mode control method based on the cerebella neural network is proposed to solve the problem that the robustness of the traditional control method is poor in view of the problem of the synchronous error of the double motor in the lifting process of the crane. On the basis of the master and slave control, the outlet pressure of the hydraulic pump and the angle of the hook are simulated as the control index. Finally, the experimental study is carried out on the lifting condition. The results show that, compared with the traditional method, the proposed control strategy can effectively improve the synchronisation control precision of two motors, strong anti-interference ability and good robust performance.

Real-time fault prevention and diagnosis in hydraulic systems is one of the challenges in engineering applications. In order to solve the problems of feature extraction and pattern recognition of different types of fault signals such as leakage, blockage, and cavitation in the hydraulic system, this paper presents a fault diagnosis method based on a multi-feature fusion support vector machine. This method first extracts the typical features of the fault signal from the time domain, frequency domain, and time−frequency domain. Then use the information entropy to calculate the weight of each feature, select the feature with the larger weight to participate in the stock index diagnosis. Finally, a multi-classifier based on the support vector machine is used to realise the fault diagnosis of the hydraulic system. The experimental results show that this method can achieve high accuracy in the hydraulic system fault diagnosis when the Gaussian radial basis function is selected. Also, compared with the commonly used improved back-projection neural network fault diagnosis method, this method has more excellent diagnostic performance.

Porcine epidemic diarrhea virus (PEDV) causes a porcine disease associated with swine epidemic diarrhea. The type I interferon (IFN-I or IFN α/β) is a key mediator of innate antiviral response during virus infection. Different antagonistic strategies have been identified and determined as to how PEDV infection inhibits the host’s IFN responses to escape the host innate immune pathway, but the pathogenic mechanisms of PEDV infection are not fully elucidated. Our preliminary results revealed that endogenous TANK-binding kinase 1 (TBK1) and interferon regulatory factor 3 (IRF3), the key components in the IFN signaling pathway were downregulated in PEDV infected IPEC-J2 cells by iTRAQ analysis. In this study, we screened nsp15 as the most important viral encoded protein involved in TBK1 and IRF3 reduction. Endoribonuclease (EndoU) activity has been well determined for coronavirus nsp15. Three residues (H226, H241, and K282) of PEDV nsp15 were identified as critical amino acids for PEDV EndoU but not D265, which was not well correlated with published results of other coronaviruses, such as severe acute respiratory syndrome virus (SARS-CoV). Moreover, PEDV nsp15 can directly degrade the RNA levels of TBK1 and IRF3 dependent on its EndoU activity to suppress IFN production and constrain the induction of IFN stimulated genes (ISGs), by which PEDV antagonizes the host innate response to facilitate its replication. Collectively, these results have confirmed that PEDV nsp15 was capable of subverting the IFN response by the RNA degradation of TBK1 and IRF3.

Porcine epidemic diarrhea (PED) re-emerged in China in 2010 and is now widespread. Evidence indicates that highly virulent porcine epidemic diarrhea virus (PEDV) strains belonging to genotype G2 caused a large-scale outbreak of diarrhea. Currently, vaccines derived from PEDV classical strains do not effectively prevent infection by virulent PEDV strains, and no specific drug is available to treat the disease. RNA interference (RNAi) is a novel and effective way to cure a wide range of viruses. We constructed three short hairpin RNA (shRNA)-expressing plasmids (shR-N307, shR-N463, and shR-N1071) directed against nucleocapsid (N) and determined their antiviral activities in intestine epithelial cells infected with a classical CV777 strain and LNCT2. We verified that shR-N307, shR-N463, and shR-N1071 effectively inhibited the expression of the transfected N gene in vitro, comparable to the control shRNA. We further demonstrated the shRNAs markedly reduced PEDV CV777 and LNCT2 replication upon downregulation of N production. Therefore, this study provides a new strategy for the design of antiviral methods against coronaviruses by targeting their processivity factors.

Alginate as a good drug delivery vehicle has excellent biocompatibility and biodegradability. In the ionic gelation process between alginate and Ca2+, the violent reaction is the absence of a well-controlled strategy in the synthesizing calcium alginate (CaA) microgels. In this study, a concentration-controlled microfluidic chip with central buffer flow was designed and 3D-printed to well-control the synthesis process of CaA microgels by the diffusion mixing pattern. The diffusion mixing pattern in the microfluidic chip can slow down the ionic gelation process in the central stream. The particle size can be influenced by channel length and flow rate ratio, which can be regulated to 448 nm in length and 235 nm in diameter. The delivery ratio of Doxorubicin (Dox) in CaA microgels are up to 90% based on the central stream strategy. CaA@Dox microgels with pH-dependent release property significantly enhances the cell killing rate against human breast cancer cells (MCF-7). The diffusion mixing pattern gives rise to well-controlled synthesis of CaA microgels, serving as a continuous and controllable production process for advanced drug delivery systems.