An Expeditious Neutralization Assay for Porcine Reproductive and Respiratory Syndrome Virus Based on a Recombinant Virus Expressing Green Fluorescent Protein.

Due to the extensive genetic and antigenic variation in Porcine Reproductive and Respiratory Syndrome Virus (PRRSV), as well as its rapid mutability and evolution, PRRS prevention and control can be challenging. An expeditious and sensitive neutralization assay for PRRSV is presented to monitor neutralizing antibodies (NAbs) in serum during vaccine research. Here, a PRRSV expressing eGFP was successfully rescued with reverse genetics based on the infectious clone HuN4-F112-eGFP which we constructed. The fluorescent protein expressions of the reporter viruses remained stable for at least five passages. Based on this reporter virus, the neutralization assay can be easily used to evaluate the level of NAbs by counting cells with green fluorescence. Compared with the classical CPE assay, the newly developed assay increases sensitivity by one- to four-fold at the early antibody response stage, thus saving 2 days of assay waiting time. By using this assay to unveil the dynamics of neutralizing antibodies against PRRSV, priming immunity through either a single virulent challenge or only vaccination could produce limited NAbs, but re-infection with PRRSV would induce a faster and stronger NAb response. Overall, the novel HuN4-F112-eGFP-based neutralization assay holds the potential to provide a highly efficient platform for evaluating the next generation of PRRS vaccines.

PSMB1 Inhibits the Replication of Porcine Reproductive and Respiratory Syndrome Virus by Recruiting NBR1 To Degrade Nonstructural Protein 12 by Autophagy.

The nonstructural proteins (Nsps) of porcine reproductive and respiratory syndrome virus (PRRSV) play essential roles in virus replication-a multistep process that requires the participation of host factors. It is of great significance for the development of antiviral drugs to characterize the host proteins that interact with PRRSV Nsps and their functions in PRRSV replication. Here, we determined that proteasome subunit ß type 1 (PSMB1) interacted with viral Nsp12 to inhibit PRRSV replication in target and permissive cells. PSMB1 could be downregulated by PRRSV infection through interaction with the transcription factor EBF1. Proteasome and autophagy inhibitor assays showed that PSMB1 was regulated by the autophagic pathway to degrade Nsp12. Cotransfection of PSMB1 and Nsp12 increased the level of intracellular autophagy; both molecules were colocated in lysosomes. We also found that the selective autophagy cargo receptor protein NBR1 and E3 ubiquitin ligase STUB1 interacted with PSMB1 and Nsp12, respectively, in the autophagic degradation of Nsp12. Furthermore, the degradation of Nsp12 by PSMB1 was mainly dependent on the ubiquitination of Nsp12 at lysine site 130. Our results indicate for the first time that PSMB1 is an anti-PRRSV host protein that inhibits the replication of PRRSV by degradation of Nsp12 through the selective autophagy pathway. IMPORTANCE PRRS is a major threat to the global pig industry and urgently requires an effective and sustainable control strategy. PRRSV Nsps have important roles in viral RNA synthesis, proteinase activity, induction of replication-associated membrane rearrangements, replicative endoribonuclease activity, determination of virulence, and regulation of host immune response. Research associated with PRRSV Nsps can provide vital guidance to modify the PRRSV genome through reverse genetics in the development of vaccines and diagnostics. The function of Nsp12, which generally plays essential roles in virus replication, remains unclear. We demonstrated that PSMB1 interacted with and degraded Nsp12 through an autophagic pathway to inhibit PRRSV replication. Our data confirmed a novel antiviral function of PSMB1 and allowed us to elaborate on the roles of Nsp12 in PRRSV pathogenesis. These findings suggest a valid and highly conserved candidate target for the development of novel therapies and more effective vaccines and demonstrate the complex cross talk between selective autophagy and PRRSV infection.

Comparison of 755-nm picosecond alexandrite laser versus 1064-nm Q-switched Nd:YAG laser for melasma: A randomized, split-face controlled, 2-year follow-up study.

OBJECTIVES: Pulsed laser treatment of melasma has shown some promising results. To compare the effectiveness and safety of 755-nm picosecond alexandrite laser (PSAL) fitted with diffractive lens array (DLA) versus 1064-nm Q-switched neodynimum:yttrium aluminum garnet laser (QSNYL) for the treatment of melasma. METHODS: We conducted a randomized, split face controlled, 2-year follow-up study. Each face was divided into two parts, each side receiving three treatments with either PSAL or QSNYL at 1 month intervals. Modified Melasma Area Severity Index scores (mMASI), pain scores, patient satisfaction and adverse events were recorded. In vivo reflectance confocal microscopy (RCM) images were acquired. RESULTS: Twenty subjects were enrolled and three dropped out. At 6 months, mMASI scores were significantly lower than baseline for QSNYL sides (p = 0.022), with no statistically significant difference between PSAL sides before and after treatment, PSAL sides versus QSNYL sides, or patient satisfaction scores. QSNYL treatment was associated with less pain (p = 0.014). No serious adverse events were reported. In the PSAL sides RCM showed a large number of dendritic melanocytes infiltrated in the dermis at 2 weeks and 4 weeks after treatment. Ten patients (58.82%) reported recurrence or exacerbation at 2-year follow-up with no statistically significant difference between the two lasers. CONCLUSIONS: QSNYL demonstrated short term clinical efficacy for melasma, but did not provide any additional benefit compared to PSAL with DLA. QSNYL was associated with less pain. There was a high recurrence rate at 2-year follow-up. RCM allowed the detection of cellular changes in melasma lesions.

Enhanced removal of nano-oil droplets utilizing polysilicate aluminum ferric (PSAF): Leveraging bridging and non-polar surface advantages.

Hydraulic oil leaks during mechanical maintenance, resulting in flushing wastewater contaminated with dispersed nano-oil droplets. In this study, 75 mg L-1 of polysilicate aluminum ferric (PSAF) was stirred at 350 rpm and the optimal chemical oxygen demand (COD) removal was 71%. The increase of PSAF led to more hydrolysis of Fe, and 1,175 cm-1 hydroxyl bridged with negative oil droplets. At the same molar concentration, PSAF hydrolyzes cationic metals more rapidly than polymeric aluminum chloride (PAC). PSAF forms flocs of smaller complex structures with greater bridging. The Al-O and Si-O peaks occurred at 611 and 1,138 cm-1, indicating the formation of Si-O-Fe and Si-O-Al bonds on the flocs surface. Higher stirring speeds did not change the free energy of the flocs surface γTot, mainly because the decrease in the van der Waals force (γLW) offset the increase of Lewis acid-base force (γAB). Preserving the non-polar surface, in summary, owing to its bridging abilities and affinity for non-polar surfaces, PSAF demonstrates superior efficiency over PAC in capturing and removing oil droplets.

The CD2v protein of African swine fever virus inhibits macrophage migration and inflammatory cytokines expression by downregulating EGR1 expression through dampening ERK1/2 activity.

African swine fever virus (ASFV) is a highly contagious and deadly virus that leads to high mortality rates in domestic swine populations. Although the envelope protein CD2v of ASFV has been implicated in immunomodulation, the molecular mechanisms underlying CD2v-mediated immunoregulation remain unclear. In this study, we generated a stable CD2v-expressing porcine macrophage (PAM-CD2v) line and investigated the CD2v-dependent transcriptomic landscape using RNA-seq. GO terms enrichment analysis and gene set enrichment analysis revealed that CD2v predominantly affected the organization and assembly process of the extracellular matrix. Wound healing and Transwell assays showed that CD2v inhibited swine macrophage migration. Further investigation revealed a significant decrease in the expression of transcription factor early growth response 1 (EGR1) through inhibiting the activity of extracellular signal-regulated kinase 1 and 2 (ERK1/2). Notably, EGR1 knockout in swine macrophages restricted cell migration, whereas EGR1 overexpression in PAM-CD2v restored the ability of macrophage migration, suggesting that CD2v inhibits swine macrophage motility by downregulating EGR1 expression. Furthermore, we performed chromatin immunoprecipitation and sequencing for EGR1 and the histone mark H3K27 acetylation (H3K27ac), and we found that EGR1 co-localized with the activated histone modification H3K27ac neighboring the transcriptional start sites. Further analysis indicated that EGR1 and H3K27ac co-occupy the promoter regions of cell locomotion-related genes. Finally, by treating various derivatives of swine macrophages with lipopolysaccharides, we showed that depletion of EGR1 decreased the expression of inflammatory cytokines including TNFα, IL1α, IL1ß, IL6, and IL8, which play essential roles in inflammation and host immune response. Collectively, our results provide new insights into the immunomodulatory mechanism of ASFV CD2v.

Diagnosis of breast cancer lesion using ultrasound images, elastography, and Ki-67 protein cell proliferation index.

Today, using elastography and ultrasound images is the best method for diagnosing breast cancer for dense tissues, especially for women under 30 years old, which is used to detect the exact border of masses. Besides, using quantitative microscopic criteria that are less tasteful seems to be useful in predicting the behavior of the tumor and its prognosis. Ki-67 is an antigen corresponding to a nuclear non-histone protein produced by cells in proliferative phases. In this article, ultrasound and elastography images of patients were collected, and breast masses were identified. The proposed algorithm includes pre-processing, feature extraction, and classification. To remove the speckle noise, two pre-processing steps are used, and after segmenting each data with its appropriate color channel, statistical features and features based on the morphology of suspicious areas are extracted. Also, sections of paraffin blocks of samples fixed in formalin were prepared and stained by immunohistochemical staining with Ki-67 monoclonal antibody, and the cell proliferation index was determined in the prepared slides. The relationship between Ki-67 positivity and microscopic grade was studied. The feature extraction results show that elastography is chosen as a more appropriate method than ultrasound due to the separation in terms of color channels. The most appropriate proposed combined methods, namely RBF-Kmeans, MLP-SCG, and RBF-SOM, have been used to classify features. The combined MLP-SCG classifier with an average accuracy of 96% and an average of 98% has improved significantly compared to other methods.

Systematic review: interventions to quit tobacco products for young adults.

BACKGROUND: Young adulthood is an important period for smoking cessation; however, there is limited evidence of smoking-cessation interventions for young adults. The aims of this study were to identify evidence-based smoking-cessation strategies for young adults, examine gaps in the literature regarding smoking cessation among young adults, and discuss methodological issues/challenges related to smoking-cessation studies for young adults. METHODS: Studies tested interventions for smoking cessation among young adults (18 to 26 years old), excluding pilot studies. Five main search engines were used, including PubMed, the Cumulative Index of Nursing and Allied Health Literature (CINAHL), EMBASE, PsycINFO, and Web of Science. The search was conducted for articles published from January 2009 to December 2019. Intervention characteristics and cessation outcomes were reviewed, and methodological quality was evaluated. RESULTS: A total of 14 articles met inclusion criteria, including randomized controlled studies and repeated cross-sectional studies. Interventions included the following: text messaging (4/14, 28.6%), social media use (2/14, 14.3%), web-or app-based intervention (2/14, 14.3%), telephone counseling (1/14, 7.1%), in-person counseling (3/14, 21.4%), pharmacological (1/14, 7.1%), and self-help booklet (1/14, 7.1%). The intervention duration and frequency of contact with participants differed and yielded varied outcomes. CONCLUSIONS: Multiple interventions have been examined to aid young adults in achieving smoking cessation. While several approaches seem promising, at the present time, the published literature is inconclusive about the type of intervention that is most effective for young adults. Future studies should compare the relative effectiveness of these intervention modalities.

An update on fractional picosecond laser treatment: histology and clinical applications.

Picosecond lasers have a very short pulse duration and a high peak power density. When fractional optical delivery systems are attached to picosecond lasers, they generate an array of concentrated microspots with a high fluence surrounded by areas with a low fluence. This article discusses the histologic characteristics and clinical applications of fractional picosecond laser treatment. Fractional picosecond laser produces laser-induced optical breakdown (LIOB) and laser-induced cavitation (LIC) in the epidermis and dermis respectively, and can encourage skin regeneration and dermal remodeling. It has been shown that fractional picosecond laser has a positive effect on facial photoaging, enlarged facial pores, dyspigmentation, wrinkles, and atrophic scars. Further research is still needed to confirm the benefits of fractional picosecond lasers.

Design of a Reconfigurable Ultra-Wideband Terahertz Polarization Rotator Based on Graphene Metamaterial.

In this work, a reconfigurable ultra-wideband transmissive terahertz polarization rotator based on graphene metamaterial is proposed that can switch between two states of polarization rotation within a broad terahertz band by changing the Fermi level of graphene. The proposed reconfigurable polarization rotator is based on a two-dimensional periodic array of multilayer graphene metamaterial structure, which is composed of metal grating, graphene grating, silicon dioxide thin film, and a dielectric substrate. The graphene metamaterial can achieve high co-polarized transmission of a linearly polarized incident wave at the off-state of the graphene grating without applying the bias voltage. Once the specially designed bias voltage is applied to change the Fermi level of graphene, the polarization rotation angle of linearly polarized waves is switched to 45° by the graphene metamaterial at the on-state. The working frequency band with 45-degree linear polarized transmission remaining above 0.7 and the polarization conversion ratio (PCR) above 90% is from 0.35 to 1.75 THz, and the relative bandwidth reaches 133.3% of the central working frequency. Furthermore, even with oblique incidence at large angles, the proposed device retains high-efficiency conversion in a broad band. The proposed graphene metamaterial offers a novel approach for the design of a terahertz tunable polarization rotator and is expected to be applied in the applications of terahertz wireless communication, imaging, and sensing.

Potential Threats to Human Health from Eurasian Avian-Like Swine Influenza A(H1N1) Virus and Its Reassortants.

During 2018-2020, we isolated 32 Eurasian avian-like swine influenza A(H1N1) viruses and their reassortant viruses from pigs in China. Genomic testing identified a novel reassortant H3N1 virus, which emerged in late 2020. Derived from G4 Eurasian H1N1 and H3N2 swine influenza viruses. This virus poses a risk for zoonotic infection.

Prediction of all-cause mortality with malnutrition assessed by nutritional screening and assessment tools in patients with heart failure：a systematic review.

BACKGROUND AND AIMS: In the absence of a gold standard or scientific consensus regarding the nutritional evaluation of heart failure (HF) patients, this study aimed to summarize and systematically evaluate the prognostic value of nutritional screening and assessment tools used for all-cause mortality in HF patients. METHODS AND RESULTS: Relevant studies were retrieved from major databases (PubMed, Embase, Web of Science, China National Knowledge Infrastructure (CNKI), WanFang Data, and China Biology Medicine disc (CMB)) and searched from the earliest available date until July 2021. If three or more studies used the same tool, meta-analysis using RevMan 5.3 was performed. This systematic review was registered at PROSPERO (number CRD42021275575). A total of 36 articles involving 25,141 HF patients were included for qualitative analysis and 31 studies for quantitative analysis. Meta-analysis of these studies indicated, poor nutritional status evaluated by using 5 nutritional screening tools (Prognostic Nutritional Index (PNI), Geriatric Nutritional Risk Index (GNRI), Controlling Nutritional Status Score (CONUT), Nutritional Risk Index (NRI), and Short Form Mini Nutritional Assessment (MNA-SF)) or 2 nutritional assessment tools (the Mini Nutritional Assessment (MNA) and Generated Subjective Global Assessment (SGA)) predicted all-cause mortality in HF patients. Of all tools analyzed, MNA had the maximum HR for mortality [HR = 2.62, 95%CI 1.11-6.20, P = 0.03] and MNA-SF [HR = 1.94, 95%CI 1.40-2.70, P＜0.001] was the best nutritional screening tools. CONCLUSION: Poor nutritional status predicted all-cause mortality in HF patients. MNA may be the best nutritional assessment tool, and MNA-SF is most recommended for HF patient nutritional screening. The application value of MNA, especially in patients with reduced left ventricular ejection fraction (LVEF), needs to be further confirmed. The clinical application value of Mini-Nutrition Assessment Special for Heart Failure (MNA-HF) and Global Leadership Initiative on Malnutrition (GLIM) in HF patients needs to be confirmed.

Instruments to measure e-cigarette related constructs: a systematic review.

BACKGROUND: Electronic cigarettes (e-cigarettes) are relatively new tobacco products that are attracting public attention due to their unique features, especially their many flavor options and their potential as an alternative to cigarettes. However, uncertainties remain regarding the determinants and consequences of e-cigarette use because current research on e-cigarettes is made more difficult due to the lack of psychometrically sound instruments that measure e-cigarette related constructs. This systematic review therefore seeks to identify the instruments in the field that are designed to assess various aspects of e-cigarette use or its related constructs and analyze the evidence presented regarding the psychometric properties of the identified instruments. METHODS: This systematic review utilized six search engines: PubMed, Medline, CINAHL, PsycINFO, Web of Science, and EMBASE, to identify articles published in the peer-reviewed journals from inception to February 2022 that contained development or validation processes for these instruments. RESULTS: Eighteen articles describing the development or validation of 22 unique instruments were identified. Beliefs, perceptions, motives, e-cigarette use, and dependence, were the most commonly assessed e-cigarette related constructs. The included studies reported either construct or criterion validity, with 14 studies reporting both. Most studies did not report the content validity; for reliability, most reported internal consistencies using Cronbach's alpha, with 15 instruments reporting Cronbach's alpha > 0.70 for the scale or its subscales. CONCLUSIONS: Twenty-two instruments with a reported development or validation process to measure e-cigarette related constructs are currently available for practitioners and researchers. This review provides a guide for practitioners and researchers seeking to identify the most appropriate existing instruments on e-cigarette use based on the constructs examined, target population, psychometric properties, and instrument length. The gaps identified in the existing e-cigarette related instruments indicate that future studies should seek to extend the validity of the instruments for diverse populations, including adolescents. Instruments that explore additional aspects of e-cigarette use and e-cigarette related constructs to help build a strong theoretical background and expand our current understanding of e-cigarette use and its related constructs, should also be developed.

The Navigation System of a Logistics Inspection Robot Based on Multi-Sensor Fusion in a Complex Storage Environment.

To reliably realize the functions of autonomous navigation and cruise of logistics robots in a complex logistics storage environment, this paper proposes a new robot navigation system based on vision and multiline lidar information fusion, which can not only ensure rich information and accurate map edges, but also meet the real-time and accurate positioning and navigation in complex logistics storage scenarios. Simulation and practical verification showed that the robot navigation system is feasible and robust, and overcomes the problems of low precision, poor robustness, weak portability, and difficult expansion of the mobile robot system in a complex environment. It provides a new idea for inspection in an actual logistics storage scenario and has a good prospective application.

Incidence of workplace violence against nurses among Chinese hospitals: A meta-analysis.

AIMS: To explore the incidence of workplace violence against nurses in Chinese hospitals. BACKGROUND: Previous systematic reviews on the incidence of workplace violence against Chinese health care workers did not include many articles published in Chinese. Although several studies have investigated cases of violence against health care providers in China, no meta-analysis has been conducted to assess the incidence of violence against Chinese nurses. EVALUATION: In this study, relevant data were retrieved from studies published up to July 2020. A meta-analysis was conducted using R software (Version 4.0). KEY FINDINGS: The 12-month incidence of workplace violence among Chinese nurses was 71% (95% CI 67%-75%), and verbal violence was the most common sub-type of violence (63%, 95% CI 58%-67%). CONCLUSION: Chinese nurses are at a high risk of violence at workplace. Hospital managers should explore ways to reduce violence against their employees, especially the younger nurses who work in secondary hospitals. IMPLICATIONS FOR NURSING MANAGEMENT: The findings of this study highlight the need to enhance the legal system in terms of laws meant to effectively mitigate violence against nurses in Chinese hospitals. Measures should be particularly taken to protect younger nurses who work in secondary hospitals.

Platelet-expressed immune checkpoint regulator GITRL in breast cancer.

Owing to their key role in several diseases including cancer, activating and inhibitory immune checkpoint molecules are increasingly exploited as targets for immunotherapy. Recently, we demonstrated that platelets, which largely influence tumor progression and immune evasion, functionally express the ligand of the checkpoint molecule GITR. This immunoreceptor modulates effector functions of T cells and NK cells with its function varying dependent on cellular context and activation state. Here, we provide a comparative analysis of platelet-derived GITRL (pGITRL) in breast cancer patients and healthy volunteers. The levels of pGITRL were found to be higher on platelets derived from cancer patients and appeared to be specifically regulated during tumor progression as exemplified by several clinical parameters including tumor stage/grade, the occurrence of metastases and tumor proliferation (Ki67) index. In addition, we report that pGITRL is upregulated during platelet maturation and particularly induced upon exposure to tumor-derived soluble factors. Our data indicate that platelets modulate the GITR/GITRL immune checkpoint in the context of malignant disease and provide a rationale to further study the GITR/GITRL axis for exploitation for immunotherapeutic intervention in cancer patients.

Risk prediction models based on hematological/body parameters for chemotherapy-induced adverse effects in Chinese colorectal cancer patients.

PURPOSE: To determine risk factors and develop novel prediction models for chemotherapy-induced adverse effects (CIAEs) in Chinese colorectal cancer (CRC) patients receiving capecitabine. METHODS: A total of 233 Chinese CRC patients receiving post-operative chemotherapy with capecitabine were randomly divided into a training set (70%) and a validation set (30%). CIAE-related hematological/body parameters were screened by univariate logistic regression. Based on a set of factors selected from LASSO (least absolute shrinkage and selection operator) logistic regression, stepwise multivariate logistic regression was applied to develop prediction models. Area under the receiver operating characteristic (ROC) curve and Hosmer-Lemeshow (HL) test were used to evaluate the discriminatory ability and the goodness of fit of each model. RESULTS: In total, 35 variables were identified to be associated with CIAEs in univariate analysis. Developed multivariable models had AUCs (area under curve) ranging from 0.625 to 0.888 and 0.428 to 0.760 in the training and validation set, respectively. The grade ≥ 1 anemia multivariable model achieved the best discriminatory ability with AUC of 0.760 (95%CI: 0.609-0.912) and good calibration with HL P value of 0.450. Then, a nomogram was constructed to predict grade ≥ 1 anemia, which included variables of age, pre-operative hemoglobin count, and pre-operative albumin count, with C-indexes of 0.775 and 0.806 in the training and validation set, respectively. CONCLUSIONS: This study identified valuable hematological/body parameters related to CIAEs. A nomogram based on the multivariable model including three hematological/body predictors can accurately predict grade ≥ 1 anemia, facilitating clinicians to implement personalized medicine early for Chinese CRC patients receiving post-operative chemotherapy for better safety treatment. Trial registration This study was registered as a clinical trial at www.clinicaltrials.gov (NCT03030508).

Transcriptional response of Bacillus megaterium FDU301 to PEG200-mediated arid stress.

BACKGROUND: For microorganisms on a paper surface, the lack of water is one of the most important stress factors. A strain of Bacillus megaterium FDU301 was isolated from plaques on a paper surface using culture medium with polyethylene glycol 200 (PEG200) to simulate an arid condition. Global transcriptomic analysis of B. megaterium FDU301 grown under normal and simulated arid conditions was performed via RNA-seq technology to identify genes involved in arid stress adaptation. RESULTS: The transcriptome of B. megaterium FDU301 grown in LB medium under arid (15% PEG200 (w/w)) and normal conditions were compared. A total of 2941 genes were differentially expressed, including 1422 genes upregulated and 1519 genes downregulated under arid conditions. Oxidative stress-responsive regulatory genes perR, fur, and tipA were significantly upregulated, along with DNA protecting protein (dps), and catalase (katE). Genes related to Fe2+ uptake (feoB), sporulation stage II (spoIIB, spoIIE, spoIIGA), small acid-soluble spore protein (sspD), and biosynthesis of compatible solute ectoine (ectB, ectA) were also highly expressed to various degrees. Oxidative phosphorylation-related genes (atpB, atpE, atpF, atpH, atpA, atpG, atpD, atpC) and glycolysis-related genes (pgk, tpiA, frmA) were significantly downregulated. CONCLUSION: This is the first report about transcriptomic analysis of a B. megaterium to explore the mechanism of arid resistance. Major changes in transcription were seen in the arid condition simulated by PEG200 (15%), with the most important one being genes related to oxidative stress. The results showed a complex mechanism for the bacteria to adapt to arid stress.

Acidity/Alkalinity of Japanese Encephalitis Virus E Protein Residue 138 Alters Neurovirulence in Mice.

The Japanese encephalitis virus (JEV) envelope (E) protein, as one of mediators of virus entry into host cells, plays a critical role in determining virulence. The Glu-to-Lys mutation of residue 138 in E protein (E138) plays an important role in attenuating JEV vaccine strain SA14-14-2. However, it is not clear how E138 attenuates JEV. Here, we demonstrate that the Glu-to-Arg mutation of E138 also determines the attenuation of JEV strain 10S3. Likewise, for its parent strain (HEN0701), a virulence strain, the mutations of E138 are responsible for virulence alteration. Furthermore, we demonstrated that mutations of alkaline residues in E138 contributed to the attenuation of neurovirulence; in contrast, mutations of acidic residues enhanced the neurovirulence of the strains. Moreover, acidity in residue E47 had a similar effect on neurovirulence. Furthermore, the alkaline E138 residue enhanced susceptibility to heparin inhibition in vitro and limited JEV diffusion in mouse brain. These results suggest that the acidity/alkalinity of the E138 residue plays an important role in neurovirulence determination.IMPORTANCE The E protein is the only glycoprotein in mature JEV, and it plays an important role in viral neurovirulence. E protein mutations attenuate JEV neurovirulence through unclear mechanisms. Here, we discovered that E138 is a predominant determinant of JEV neurovirulence. We demonstrated that the alkalinity/acidity of E138 determines JEV neurovirulence. These data contribute to the characterization of the E protein and the rational development of novel JEV vaccines.

Sema3E is required for migration of cranial neural crest cells in zebrafish: Implications for the pathogenesis of CHARGE syndrome.

CHARGE syndrome is a congenital disorder with multiple malformations in the craniofacial structures, and cardiovascular and genital systems, which are mainly affected by neural crest defects caused by loss-of-function mutations within chromodomain helicase DNA-binding protein 7 (CHD7). However, many patients with CHARGE syndrome test negative for CHD7. Semaphorin 3E (sema3E) is a gene reported to be mutated in patients with CHARGE syndrome. However, its role in the pathogenesis of CHARGE syndrome has not been verified experimentally. Here, we report that the knockdown of sema3E results in severe craniofacial malformations, including small eyes, defective cartilage and an abnormal number of otoliths in zebrafish embryos, which resemble the major features of CHARGE syndrome. Further analysis reveals that the migratory cranial neural crest cells are scattered in the region of the hindbrain, and the postmigratory neural crest cells are reduced in the pharyngeal arches upon sema3E knockdown. Notably, immunostaining and time-lapse imaging analyses of a neural crest cell-labelled transgenic fish line, sox10:EGFP, show that the migration of cranial neural crest cells is severely impaired, and many of these cells are misrouted upon sema3E knockdown. Furthermore, the sox10-expressing cranial neural crest cells are scattered in chd7 homozygous mutants, which phenocopied the phenotype in sema3E morphants. Overexpression of sema3E rescues the phenotype of scattered cranial neural crest cells in chd7 homozygotes, indicating that chd7 may control the expression of sema3E to regulate cranial neural crest cell migration. Collectively, our data demonstrate that sema3E is involved in the pathogenesis of CHARGE syndrome by modulating cranial neural crest cell migration.

MOV10 inhibits replication of porcine reproductive and respiratory syndrome virus by retaining viral nucleocapsid protein in the cytoplasm of Marc-145 cells.

Porcine reproductive and respiratory syndrome virus (PRRSV) has been a major threat to global industrial pig farming ever since its emergence in the late 1980s. Identification of sustainable and effective control measures against PRRSV transmission is a pressing problem. The nucleocapsid (N) protein of PRRSV is specifically localized in the cytoplasm and nucleus of virus-infected cells which is important for PRRSV replication. In the current study, a new host restricted factor, Moloney leukemia virus 10-like protein (MOV10), was identified as an inhibitor of PRRSV replication. N protein levels and viral replication were significantly reduced in Marc-145 cells stably overexpressing MOV10 compared with those in wild-type Marc-145 cells. Adsorption experiments revealed that MOV10 did not affect the attachment and internalization of PRRSV. Co-immunoprecipitation and immunofluorescence co-localization analyses showed that MOV10 interacted and co-localized with the PRRSV N protein in the cytoplasm. Notably, MOV10 affected the distribution of N protein in the cytoplasm and nucleus, leading to the retention of N protein in the former. Taken together, these findings demonstrate for the first time that MOV10 inhibits PRRSV replication by restricting the nuclear import of N protein. These observations have great implications for the development of anti-PRRSV drugs and provide new insight into the role of N protein in PRRSV biology.