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Abstract Marek’s disease virus (MDV) is a highly pathogenic and oncogenic alpha herpesvirus that causes Marek’s disease (MD), which is one of the most important immunosuppressive and rapid-onset neoplastic diseases in poultry. The onset of MD lymphomas and other clinical diseases can be efficiently prevented by vaccination; these vaccines are heralded as the first demonstration of a successful vaccination strategy against a cancer. However, the persistent evolution of epidemic MDV strains towards greater virulence has recently resulted in frequent outbreaks of MD in vaccinated chicken flocks worldwide. Herein, we provide an overall review focusing on the discovery and identification of the strategies by which MDV evades host immunity and attacks the immune system. We have also highlighted the decrease in the immune efficacy of current MD vaccines. The prospects, strategies and new techniques for the development of efficient MD vaccines, together with the possibilities of antiviral therapy in MD, are also discussed.

In the past decade, research has demonstrated that viral miRNAs encoded by a number of viral genomes, particularly by most of the herpesvirus including Marek's disease virus (MDV), play important regulatory roles in viral infection, replication, and regulation of tumorigenesis. As macrovesicles in cells, exosomes can deliver viral miRNAs and exert gene regulatory functions. Whether the exosomes play a role in the replication, pathogenesis/tumorigenesis of avian herpesviruses such as oncogenic Marek's disease virus (MDV) remains unclear. Herein we extracted and identified the exosomes from MDV-transformed T cell line MSB-1 and demonstrated high abundance of MDV-1 miRNA expression. Using dual luciferase-based reporter assay, we also demonstrated that the exosomes derived from MSB-1 can deliver functional miRNA successfully into primary chicken embryo fibroblasts. These findings provide new insights into the role of exosomes and the mechanisms of how virus-encoded miRNA function in MDV latency/activation switching, viral replication, pathogenesis and/or tumorigenesis.

Abstract Marek’s disease virus (MDV) strain GX0101 was the first reported field strain of recombinant gallid herpesvirus type 2 (GaHV-2). However, the splenic proteome of MDV-infected chickens remains unclear. In this study, a total of 28 1-day-old SPF chickens were intraperitoneally injected with chicken embryo fibroblast (CEF) containing 2000 PFU GX0101. Additionally, a control group, consisting of four one-day-old SPF chickens, received intraperitoneal equal doses of CEF. Blood and various tissue samples were collected at different intervals (7, 14, 21, 30, 45, 60, and 90 days post-infection; dpi) for histopathological, real-time PCR, and label-free quantitative analyses. The results showed that the serum expressions of MDV-related genes, meq and gB, peaked at 45 dpi. The heart, liver, and spleen were dissected at 30 and 45 dpi, and their hematoxylin-eosin staining indicated that virus infection compromised the normal organizational structure at 45 dpi. Particularly, the spleen structure was severely damaged, and the lymphocytes in the white medulla were significantly reduced. Furthermore, liquid chromatography-mass spectrometry (LC-MS) and label-free techniques were used to analyze the difference in splenic proteome profiles of the experimental and control groups at 30 and 45 dpi. Proteomic analysis identified 1660 and 1244 differentially expressed proteins (DEPs) at 30 and 40 dpi, respectively, compared with the uninfected spleen tissues. According to GO analysis, these DEPs were involved in processes such as organelle organization, cellular component biogenesis, cellular component assembly, anion binding, small molecule binding, metal ion binding, cation binding, cytosol, nuclear part, etc. Additionally, KEGG analysis indicated that the following pathways were linked to MDV-induced inflammation, apoptosis, and tumor: Wnt, Hippo, AMPK, cAMP, Notch, TGF-β, PI3K-Akt, Rap1, Ras, Calcium, NF-κB, PPAR, cGMP-PKG, Apoptosis, VEGF, mTOR, FoxO, TNF, JAK-STAT, MAPK, Prion disease, T cell receptor, and B cell receptor. We finally screened 674 DEPs that were linked to MDV infection in spleen tissue. This study improves our understanding of the MDV response mechanism in the spleen.

Myocardial infarction (MI) and its associated poor prognosis pose significant risks to human health. Nanomaterials hold great potential for the treatment of MI due to their targeted and controlled release properties, particularly biomimetic nanomaterials. The utilization of biomimetic strategies based on extracellular vesicles (EVs) and cell membranes will serve as the guiding principle for the development of nanomaterial therapy in the future. In this review, we present an overview of research progress on various exosomes derived from mesenchymal stem cells, cardiomyocytes, or induced pluripotent stem cells in the context of myocardial infarction (MI) therapy. These exosomes, utilized as cell-free therapies, have demonstrated the ability to enhance the efficacy of reducing the size of the infarcted area and preventing ischaemic reperfusion through mechanisms such as oxidative stress reduction, polarization modulation, fibrosis inhibition, and angiogenesis promotion. Moreover, EVs can exert cardioprotective effects by encapsulating therapeutic agents and can be engineered to specifically target the infarcted myocardium. Furthermore, we discuss the use of cell membranes derived from erythrocytes, stem cells, immune cells and platelets to encapsulate nanomaterials. This approach allows the nanomaterials to camouflage themselves as endogenous substances targeting the region affected by MI, thereby minimizing toxicity and improving biocompatibility. In conclusion, biomimetic nano-delivery systems hold promise as a potentially beneficial technology for MI treatment. This review serves as a valuable reference for the application of biomimetic nanomaterials in MI therapy and aims to expedite the translation of NPs-based MI therapeutic strategies into practical clinical applications.

Abstract Female solo travel is experiencing a global increase and specifically, gaining popularity in Asia. This study explores how personal values and female solo travel motivation affect travel behavior. Using a sample comprising 381 single females in Taiwan, partial least squares structural equation modeling was utilized to investigate the hypotheses. The results revealed Hypothesis 1 and Hypothesis 3 are supported, which verifies personal internal values significantly affect female solo travel motivation, and are identified as significant factors influencing female solo travel intention. Additionally, Hypothesis 5 is partially support, indicating the female solo travel motivations of escape/relaxation, relationship, and self-actualization contribute to the formation of positive female solo travel intention. As Hypothesis 2 and Hypothesis 4 are unsupported, external values have no impact on female solo travel motivation or any significant effect on female solo travel intention. This research adds to the vast gap in tourism literature by identifying the personal values and motivations of female solo travel, and benefits the development of the female solo travel market.

ABSTRACT Marek’s disease virus (MDV) induces immunosuppression and neoplastic disease in chickens. The virus is controllable via an attenuated meq deletion mutant virus, which has the disadvantage of retaining the ability to induce lymphoid organ atrophy. To overcome this deficiency and produce more vaccine candidates, a recombinant MDV was generated from the highly virulent Md5BAC strain, in which both meq and a cytolytic replication-related gene, pp38, were deleted. Replication of the double deletion virus, Md5BAC ΔmeqΔpp38, was comparable with that of the parental virus in vitro. The double deletion virus was shown to be fully attenuated and to reduce lymphoid organ atrophy in vivo. Crucially, Md5BAC ΔmeqΔpp38 confers superior protection against highly virulent virus compared with a commercial vaccine strain, CVI988/Rispens. Transcriptomic profiling indicated that Md5BAC ΔmeqΔpp38 induced a different host immune response from CVI988/Rispens. In summary, a novel, effective, and safe vaccine candidate for prevention and control of MD caused by highly virulent MDV is reported. IMPORTANCE MDV is a highly contagious immunosuppressive and neoplastic pathogen. The virus can be controlled through vaccination via an attenuated meq deletion mutant virus that retains the ability to induce lymphoid organ atrophy. In this study, we overcame the deficiency by generating meq and pp38 double deletion mutant virus. Indeed, the successfully generated meq and pp38 double deletion mutant virus had significantly reduced replication capacity in vivo but not in vitro. It was fully attenuated and conferred superior protection efficacy against very virulent MDV challenge. In addition, the possible immunological protective mechanism of the double deletion mutant virus was shown to be different from that of the gold standard MDV vaccine, CVI988/Rispens. Overall, we successfully generated an attenuated meq deletion mutant virus and widened the range of potential vaccine candidates. Importantly, this study provides for the first time the theoretical basis of vaccination induced by fully attenuated gene-deletion mutant virus.

Due to the effect COVID-19 epidemic, promoting green consumption is now a key marketing strategy in the hospitality and tourism industry. As it is vital green hotels predict their customers’ visit intention, this study attempts to discover the factors affecting Taiwan’s Z-generation tourists’ green hotel visit intention using an extended theory of planned behavior [including personal moral norms (PMN) and environmental concern (EC)]. Data were gathered from 296 Z-generation tourists via an online survey, which was subsequently analyzed using partial least squares structural equation modeling. The results evidence that Z-generation tourists’ attitude, subjective norms, (SN) and perceived behavioral control positively and significantly influence their green hotel visit intention, with attitude being the most significant factor. Moreover, the mediation model analysis indicates Z-generation tourists’ attitude toward green hotels mediates the relationships between PMN, SN, EC, and visit intention. This study provides new insights into tourists’ green hotel visit intention and emphasizes the importance of attitude in the formation of intention.

As one of the most important avian immunosuppressive and neoplastic diseases, Marek’s disease (MD), caused by oncogenic Marek’s disease virus (MDV), has caused huge economic losses worldwide over the past five decades. In recent years, MD outbreaks have occurred frequently in MD-vaccinated chicken flocks, but the key pathogenic determinants and influencing factors remain unclear. Herein, we analyzed the pathogenicity of seven newly isolated MDV strains from tumor-bearing chickens in China and found that all of them were pathogenic to chicken hosts, among which four MDV isolates, SDCW01, HNXZ05, HNSQ05 and HNSQ01, were considered to be hypervirulent MDV (HV-MDV) strains. At 73 days of the virus infection experiment, the cumulative incidences of MD were 100%, 93.3%, 90% and 100%, with mortalities of 83.3%, 73.3%, 60% and 86.7%, respectively, for the four viruses. The gross occurrences of tumors were 50%, 33.3%, 30% and 63.3%, respectively, accompanied by significant hepatosplenomegaly and serious atrophy of the immune organs. Furthermore, the immune protection effects of four commercial MD vaccines against SDCW01, CVI988, HVT, CVI988+HVT, and 814 were explored. Unexpectedly, during the 67 days of post-virus challenge, the protection indices (PIs) of these four MD vaccines were only 46.2%, 38.5%, 50%, and 28%, respectively, and the birds that received the monovalent CVI988 or HVT still developed tumors with cumulative incidences of 7.7% and 11.5%, respectively. To our knowledge, this is the first demonstration of the simultaneous comparison of the immune protection efficacy of multiple commercial MD vaccines with different vaccine strains. Our study revealed that the HV-MDV variants circulating in China could significantly break through the immune protection of the classical MD vaccines currently widely used. For future work, there is an urgent need to develop novel, more effective MD vaccines for tackling the new challenge of emerging HV-MDV strains or variants for the sustainable control of MD.

Abstract Marek’s disease virus (MDV) induces severe immunosuppression and lymphomagenesis in the chicken, its natural host, and results in a condition that investigated the pathogenesis of MDV and have begun to focus on the expression profiling of circular RNAs (circRNAs). However, little is known about how the expression of circRNAs is referred to as Marek’s disease. Previous reports have is regulated during MDV replication. Here, we carried out a comprehensive profiling analysis of N6-methyladenosine (m6A) modification on the circRNA transcriptome in infected and uninfected chicken embryonic fibroblast (CEF) cells. Methylated RNA immunoprecipitation sequencing (MeRIP-Seq) revealed that m6A modification was highly conserved in circRNAs. Comparing to the uninfected group, the number of peaks and conserved motifs were not significantly different in cells that were infected with MDV, although reduced abundance of circRNA m6A modifications. However, gene ontology and Kyoto encyclopedia of genes and genomes (KEGG) pathway analyses revealed that the insulin signaling pathway was associated with the regulation of m6A modified circRNAs in MDV infection. This is the first report to describe alterations in the transcriptome-wide profiling of m6A modified circRNAs in MDV-infected CEF cells.

Abstract Background The newly discovered reversible N6-methyladenosine (m6A) modification plays an important regulatory role in gene expression. Long non-coding RNAs (lncRNAs) participate in Marek’s disease virus (MDV) replication but how m6A modifications in lncRNAs are affected during MDV infection is currently unknown. Herein, we profiled the transcriptome-wide m6A modification in lncRNAs in MDV-infected chicken embryo fibroblast (CEF) cells. Results Methylated RNA immunoprecipitation sequencing results revealed that the lncRNA m6A modification is highly conserved with MDV infection increasing the expression of lncRNA m6A modified sites compared to uninfected cell controls. Gene Ontology and the Kyoto Encyclopedia of Genes and Genomes pathway analysis revealed that lncRNA m6A modifications were highly associated with signaling pathways associated with MDV infection. Conclusions In this study, the alterations seen in transcriptome-wide m6A occurring in lncRNAs following MDV-infection suggest this process plays important regulatory roles during MDV replication. We report for the first time profiling of the alterations in transcriptome-wide m6A modification in lncRNAs of MDV-infected CEF cells.

Abstract Processing and packaging of herpesvirus genomic DNA is regulated by a packaging-associated terminase complex comprising of viral proteins pUL15, pUL28 and pUL33. Marek’s disease virus (MDV) homologs UL28 and UL33 showed conserved functional features with high sequence identity with the corresponding Herpes simplex virus 1 (HSV-1) homologs. As part of the investigations into the role of the UL28 and UL33 homologs of oncogenic MDV for DNA packaging and replication in cultured cells, we generated MDV mutant clones deficient in UL28 or UL33 of full-length MDV genomes. Transfection of UL28- or UL33-deleted BAC DNA into chicken embryo fibroblast (CEF) did not result either in the production of visible virus plaques, or detectable single cell infection after passaging onto fresh CEF cells. However, typical MDV plaques were detectable in CEF transfected with the DNA of revertant mutants where the deleted genes were precisely reinserted. Moreover, the replication defect of the UL28-deficient mutant was completely restored when fragment encoding the full UL28 gene was co-transfected into CEF cells. Viruses recovered from the revertant construct, as well as by the UL28 co-transfection, showed replication ability comparable with parental virus. Furthermore, the transmission electron microscopy study indicated that immature capsids were assembled without the UL28 expression, but with the loss of infectivity. Importantly, predicted three-dimensional structures of UL28 between MDV and HSV-1 suggests conserved function in virus replication. For the first time, these results revealed that both UL28 and UL33 are essential for MDV replication through regulating DNA cleavage and packaging.

Marek’s disease (MD) caused by pathogenic Marek’s disease virus type 1 (MDV−1) is one of the most important neoplastic diseases of poultry. MDV−1-encoded unique Meq protein is the major oncoprotein and the availability of Meq-specific monoclonal antibodies (mAbs) is crucial for revealing MDV pathogenesis/oncogenesis. Using synthesized polypeptides from conserved hydrophilic regions of the Meq protein as immunogens, together with hybridoma technology and primary screening by cross immunofluorescence assay (IFA) on Meq-deleted MDV−1 viruses generated by CRISPR/Cas9-gene editing, a total of five positive hybridomas were generated. Four of these hybridomas, namely 2A9, 5A7, 7F9 and 8G11, were further confirmed to secrete specific antibodies against Meq as confirmed by the IFA staining of 293T cells overexpressing Meq. Confocal microscopic analysis of cells stained with these antibodies confirmed the nuclear localization of Meq in MDV-infected CEF cells and MDV-transformed MSB-1 cells. Furthermore, two mAb hybridoma clones, 2A9-B12 and 8G11-B2 derived from 2A9 and 8G11, respectively, displayed high specificity for Meq proteins of MDV−1 strains with diverse virulence. Our data presented here, using synthesized polypeptide immunization combined with cross IFA staining on CRISPR/Cas9 gene-edited viruses, has provided a new efficient approach for future generation of specific mAbs against viral proteins.

Abstract Background Porcine circovirus type 2 (PCV2) is the pathogen of porcine circovirus associated diseases (PCVAD) and one of the main pathogens in the global pig industry, which has brought huge economic losses to the pig industry. In recent years, there has been limited research on the prevalence of PCV2 in Henan Province. This study investigated the genotype and evolution of PCV2 in this area. Results We collected 117 clinical samples from different regions of Henan Province from 2015 to 2018. Here, we found that the PCV2 infection rate of PCV2 was 62.4%. Thirty-seven positive clinical samples were selected to amplify the complete genome of PCV2 and were sequenced. Based on the phylogenetic analysis of PCV2 ORF2 and complete genome, it was found that the 37 newly detected strains belonged to PCV2a (3 of 37), PCV2b (21 of 37) and PCV2d (13 of 37), indicating the predominant prevalence of PCV2b and PCV2d strains. In addition, we compared the amino acid sequences and found several amino acid mutation sites among different genotypes. Furthermore, the results of selective pressure analysis showed that there were 5 positive selection sites. Conclusions This study indicated the genetic diversity, molecular epidemiology and evolution of PCV2 genotypes in Henan Province during 2015–2018.

The avian immunosuppressive and neoplastic diseases caused by Marek’s disease virus (MDV), avian leucosis virus (ALV), and reticuloendotheliosis virus (REV) are seriously harmful to the global poultry industry. In recent years, particularly in 2020–2022, outbreaks of such diseases in chicken flocks frequently occurred in China. Herein, we collected live diseased birds from 30 poultry farms, out of 42 farms with tumour-bearing chicken flocks distributed in central China, to investigate the current epidemiology and co-infections of these viruses. The results showed that in individual diseased birds, the positive infection rates of MDV, ALV, and REV were 69.5% (203/292), 14.4% (42/292), and 4.7% (13/277), respectively, while for the flocks, the positive infection rates were 96.7% (29/30), 36.7% (11/30), and 20% (6/30), respectively. For chicken flocks, monoinfection of MDV, ALV, or REV was 53.3% (16/30), 3.3% (1/30), and 0% (0/30), respectively, but a total of 43.3% (13/30) co-infections was observed, which includes 23.3% (7/30) of MDV+ALV, 10.0% (3/30) of MDV+REV, and 10.0% (3/30) of MDV+ALV+REV co-infections. Interestingly, no ALV+REV co-infection or REV monoinfection was observed in the selected poultry farms. Our data indicate that the prevalence of virulent MDV strains, partially accompanied with ALV and/or REV co-infections, is the main reason for current outbreaks of avian neoplastic diseases in central China, providing an important reference for the future control of disease.

Cucumber seeds are the seeds of Cucumber (Cucumis sativus L.), which include a large number of essential fatty acids, plant sterols, glycosides, volatile oils, as well as Ca, Mg, and other inorganic elements. Cucumber seed oil has good UV absorption effect because it is rich in an unsaturated functional group structure. Plant sterol in cucumber seed oil shows strong permeability toward the skin, which promotes skin metabolism and inhibits skin inflammation. β-sitosterol in plant sterols can also effectively protect the peroxide of low-density lipoprotein, so they have an antioxidant effect. In this study, cucumber seed oil was extracted by the Soxhlet extraction method as plant-based sunscreen, in which cucumber seed oil was used as the main component, and an appropriate amount of titanium dioxide was used as an antioxidant. Taking the settling solvent ratio as the index, the orthogonal experiment was designed to determine the prescription composition of the pure matrix. Then the single factor experiments of cucumber seed oil, titanium dioxide, and vitamin E were carried out. The effects of three components on UV absorption were investigated by orthogonal experiments, and the optimum formula with cucumber seed oil as the main component was finally determined and verified. In conclusion, our plant sunscreen was produced by cucumber seed oil as raw material instead of hormone. There was no harm to the skin. It has the potential to develop plant-based sunscreen with cucumber seed oil and provides a reference for the development of plant sunscreen and skin care products.

Suicide is one of the leading causes of mortality worldwide. Various factors could lead to suicidal ideation (SI), while depression is the predominant cause among all mental disorders. Studies have shown that alterations in brain structures and networks may be highly associated with suicidality. This study investigated both neurological structural variations and network alterations in depressed patients with suicidal ideation by using generalized q-sampling imaging (GQI) and Graph Theoretical Analysis (GTA). This study recruited 155 participants and divided them into three groups: 44 depressed patients with suicidal ideation (SI+; 20 males and 24 females with mean age = 42, SD = 12), 56 depressed patients without suicidal ideation (Depressed; 24 males and 32 females with mean age = 45, SD = 11) and 55 healthy controls (HC; nine males and 46 females with mean age = 39, SD = 11). Both the generalized fractional anisotropy (GFA) and normalized quantitative anisotropy (NQA) values were evaluated in a voxel-based statistical analysis by GQI. We analyzed different topological parameters in the graph theoretical analysis and the subnetwork interconnections in the Network-based Statistical (NBS) analysis. In the voxel-based statistical analysis, both the GFA and NQA values in the SI+ group were generally lower than those in the Depressed and HC groups in the corpus callosum and cingulate gyrus. Furthermore, we found that the SI+ group demonstrated higher global integration and lower local segregation among the three groups of participants. In the network-based statistical analysis, we discovered that the SI+ group had stronger connections of subnetworks in the frontal lobe than the HC group. We found significant structural differences in depressed patients with suicidal ideation compared to depressed patients without suicidal ideation and healthy controls and we also found several network alterations among these groups of participants, which indicated that white matter integrity and network alterations are associated with patients with depression as well as suicidal ideation.

Marek’s disease virus (MDV) is an important oncogenic α-herpesvirus that induces Marek’s disease (MD), characterized by severe immunosuppression and rapid-onset T-cell lymphomas in its natural chicken hosts. Historically, MD is regarded as an ideal biomedical model for studying virally induced cancers. Monoclonal antibodies (mAbs) against viral or host antigenic epitopes are crucial for virology research, especially in the exploration of gene functions, clinical therapy, and the development of diagnostic reagents. Utilizing the CRISPR/Cas9-based gene-editing technology, we produced a pp38-deleted MDV-1 mutant—GX0101Δpp38—and used it for the rapid screening and identification of pp38-specific mAbs from a pool of MDV-specific antibodies from 34 hybridomas. The cross-staining of parental and mutated MDV plaques with hybridoma supernatants was first performed by immunofluorescence assay (IFA). Four monoclonal hybridomas—namely, 4F9, 31G7, 34F2, and 35G9—were demonstrated to secrete specific antibodies against MDV-1’s pp38 protein, which was further confirmed by IFA staining and confocal analysis. Further experiments using Western blotting, immunoprecipitation (IP), liquid chromatography–tandem mass spectrometry (LC–MS/MS), and immunohistochemistry (IHC) analysis demonstrated that the pp38-specific mAb 31G7 has high specificity and wide application potential for further research in MD biology. To the best of our knowledge, this is the first demonstration of the use of CRISPR/Cas9-based gene-editing technology for efficient screening and identification of mAbs against a specific viral protein, and provides a meaningful reference for the future production of antibodies against other viruses—especially for large DNA viruses such as herpesviruses.

During the coronavirus disease 2019 (COVID-19) pandemic, quarantine hotel employees face a higher risk of infection while they host quarantine guests from overseas. This is the first research to empirically investigate the psychological effects of operating a quarantine hotel on its employees. The empirical results indicate that heightened fear of COVID-19 leads to adverse mental health issues for quarantine hotel employees and confirm that depression, anxiety, and stress have a significant influence on turnover intention. These findings contribute to the extant knowledge base by uncovering the role of mental health in employee turnover intention. Based on the results, implications are presented for practitioners.

In recent years, outbreaks of Marek’s disease (MD) have been frequently reported in vaccinated chicken flocks in China. Herein, we have demonstrated that four Marek’s disease virus (MDV) isolates, HN502, HN302, HN304, and HN101, are all pathogenic and oncogenic to hosts. Outstandingly, the HN302 strain induced 100% MD incidence, 54.84% mortality, and 87.10% tumor incidence, together with extensive atrophy of immune organs. Pathotyping of HN302 was performed in comparison to a standard very virulent (vv) MDV strain Md5. We found that both CVI988 and HVT vaccines significantly reduced morbidity and mortality induced by HN302 or Md5 strains, but the protection indices (PIs) provided by these two vaccines against HN302 were significantly lower (27.03%) or lower (33.33%) than that against Md5, which showed PIs of 59.89% and 54.29%, respectively. These data suggested that HN302 possesses a significant higher virulence than Md5 and at least could be designated as a vvMDV strain. Together with our previous phylogenetic analysis on MDV-1 meq genes, we have presently suggested HN302 to be a typical highly virulent MDV variant belonging to an independent Chinese branch. To our knowledge, this is the first report to provide convincible evidence to identify a pathogenic MDV variant strain with a higher virulence than Md5 in China, which may have emerged and circulating in poultry farms in China for a long time and involved in the recent MD outbreaks.

MicroRNAs (miRNAs) have been demonstrated for their involvement in virus biology and pathogenesis, including functioning as key determinants of virally-induced cancers. As an important oncogenic α-herpesvirus affecting poultry health, Marek’s disease virus serotype 1 [Gallid alphaherpesvirus 2 (GaHV-2)] induces rapid-onset T-cell lymphomatous disease commonly referred to as Marek’s disease (MD), an excellent biological model for the study of virally-induced cancer in the natural hosts. Previously, we have demonstrated that GaHV-2-encoded miRNAs (especially those within the Meq-cluster) have the potential to act as critical regulators of multiple processes such as virus replication, latency, pathogenesis, and/or oncogenesis. In addition to miR-M4-5p (miR-155 homolog) and miR-M3-5p, we have recently found that miR-M2-5p possibly participate in inducing MD lymphomagenesis. Here, we report the identification of two tumor suppressors, the RNA-binding protein 24 (RBM24) and myogenic differentiation 1 (MYOD1), being two biological targets for miR-M2-5p. Our experiments revealed that as a critical miRNA, miR-M2-5p promotes cell proliferation via regulating the RBM24-mediated p63 overexpression and MYOD1-mediated IGF2 signaling and suppresses apoptosis by targeting the MYOD1-mediated Caspase-3 signaling pathway. Our data present a new strategy of a single viral miRNA exerting dual role to potentially participate in the virally-induced T-cell lymphomagenesis by simultaneously promoting the cell proliferation and suppressing apoptosis.

The physical arch and the arch effect are related to many geotechnical problems, such as the stability of mine openings, a tunnel's shape, and disaster prevention. However, observing the arch in geo-material is challenging due to Earth's invisible force network distribution. The arch shape is a critical parameter related to the arching effect, which is still an open question. Specifically, the force network evolution after local excavation in granular material is revealed using the photo-elastic technique, and the system pressure variation is calculated qualitatively in particle scale according to the G2 algorithm. Besides, the force chain arch is formed by strong force chains, which are important in redistributing the stress field. The principal direction of the spatial correlation colormap represents the direction of the strong force chains in the network. Based on this preferred angle between the strong force chain and the horizontal direction, a simplified force chain arch trajectory is iteratively rebuilt. Then, the total force chain distribution is studied considering both G2 and the total length of strong force chains, with the corresponding results demonstrating a relationship between the changes in these characteristics and the force chain arch formation. [ABSTRACT FROM AUTHOR]

The clogging of a stream bed has a significant impact on the exchange of water between a river and groundwater, known as hyporheic exchange. Clogging issues in orifices influenced by fluid flow have long been a concern, particularly in understanding clogging mechanisms and particle dynamics. This study focuses on investigating the behavior of poly-dispersed particle systems in the Shanghai soft soil using a coupled lattice Boltzmann method and a discrete element method (LBM-DEM). A particle-orifice model was developed based on LBM-DEM to study the clogging problem. Three different orifice diameters with the same particle size distribution were set at a microscopic scale. The simulation results indicate that the primary form of clogging is the formation of particle domes, exhibiting distinct characteristics depending on orifice size. As the orifice size increases, clogging becomes more challenging. Interestingly, the flow rate of fluid passing through clogged orifices is quite complex, as it does not decrease significantly. The formation process of the particle structure shows significant randomness, posing challenges for quantitative analysis and experimental validation. This study provides insights into the initial stages of clogging formation, leading to a better understanding of hyporheic exchange phenomena in stream bed ecosystems. [ABSTRACT FROM AUTHOR]

During the pandemic, quarantine hotel workers face a higher risk of infection while they host quarantine guests from overseas. This study’s aim is to gain an understanding of the knowledge, attitudes, and practices (KAP) of quarantine hotel workers in China. A total of 170 participants took part in a cross-sectional survey to assess the KAP of quarantine hotel workers in China, during the COVID-19 pandemic. The chi-square test, independent t-test, one-way analysis of variance (ANOVA), descriptive analysis, and binary logistic regression were used to examine the sociodemographic factors associated with KAP levels during the COVID-19 pandemic. The results show that 62.41% have good knowledge, 94.7% have a positive attitude towards COVID-19, but only 78.2% have good practices. Most quarantine hotel workers (95.3%) are confident that COVID-19 will be successfully controlled and that China is handling the COVID-19 crisis well (98.8%). Most quarantine hotel workers are also taking personal precautions, such as avoiding crowds (80.6%) and wearing facemasks (97.6%). The results evidence that quarantine hotel workers in China have acquired the necessary knowledge, positive attitudes and proactive practices in response to the COVID-19 pandemic. The results of this study can provide a reference for quarantine hotel workers and their targeted education and intervention.

In recent years, the CRISPR/Cas9-based gene-editing techniques have been well developed and applied widely in several aspects of research in the biological sciences, in many species, including humans, animals, plants, and even in viruses. Modification of the viral genome is crucial for revealing gene function, virus pathogenesis, gene therapy, genetic engineering, and vaccine development. Herein, we have provided a brief review of the different technologies for the modification of the viral genomes. Particularly, we have focused on the recently developed CRISPR/Cas9-based gene-editing system, detailing its origin, functional principles, and touching on its latest achievements in virology research and applications in vaccine development, especially in large DNA viruses of humans and animals. Future prospects of CRISPR/Cas9-based gene-editing technology in virology research, including the potential shortcomings, are also discussed.

Hot mixed asphalt (HMA) is a mixture of particles (coarse and fine aggregates) and interstitial fluid (asphalt binder) designed to compact and harden for long-lasting roads. In this study, we implement a two-scale approach to capture the compaction behaviour of hot asphalt mixtures using both a granular-slurry rheology (GSR) at a smaller scale and a discrete element method (DEM) simulation at the scale of a compactor. We show that this modelling effort captures the compaction of HMA with different binder viscosities modified by adding graphene nano-platelets (GNP). This research has the capacity to shed light on how the properties of mixture components can influence compaction efficiency and effectiveness.

The virus-encoded microRNAs (miRNAs) have been demonstrated to have important regulatory roles in herpesvirus biology, including virus replication, latency, pathogenesis and/or tumorigenesis. As an emerging efficient tool for gene editing, the clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9 system has been successfully applied in manipulating the genomes of large DNA viruses. Herein, utilizing the CRISPR/Cas9 system with a double-guide RNAs transfection/virus infection strategy, we have established a new platform for mutagenesis of viral miRNAs encoded by the Marek’s disease virus serotype 1 (MDV-1), an oncogenic alphaherpesvirus that can induce rapid-onset T-cell lymphomas in chickens. A series of miRNA-knocked out (miR-KO) mutants with deletions of the Meq- or the mid-clustered miRNAs, namely RB-1B∆Meq-miRs, RB-1B∆M9-M2, RB-1B∆M4, RB-1B∆M9 and RB-1B∆M11, were generated from vvMDV strain RB-1B virus. Interestingly, mutagenesis of the targeted miRNAs showed changes in the in vitro virus growth kinetics, which is consistent with that of the in vivo proliferation curves of our previously reported GX0101 mutants produced by the bacterial artificial chromosome (BAC) clone and Rec E/T homologous recombination techniques. Our data demonstrate that the CRISPR/Cas9-based gene editing is a simple, efficient and relatively nondisruptive approach for manipulating the small non-coding genes from the genome of herpesvirus and will undoubtedly contribute significantly to the future progress in herpesvirus biology.

The behaviour of granular column collapses is associated with the dynamics of geohazards, such as debris flows, landslides and pyroclastic flows, yet their underlying physics is still not well understood. In this paper, we explore granular column collapses using the sphero-polyhedral discrete element method, where the system contains two types of particles with different frictional properties. We impose three different mixing ratios and multiple different particle frictional coefficients, which lead to different run-out distances and deposition heights. Based on our previous work and a simple mixture theory, we propose a new effective initial aspect ratio for the bi-frictional granular mixture, which helps unify the description of the relative run-out distances. We analyse the kinematics of bi-frictional granular column collapses and find that deviations from classical power-law scaling in both the dimensionless terminal time and the dimensionless time when the system reaches the maximum kinetic energy may result from differences in the initial solid fraction and initial structures. To clarify the influence of initial states, we further decrease the initial solid fraction of granular column collapses, and propose a trial function to quantitatively describe its influence. Due to the utilization of a simple mixture theory of contact occurrence probability, this study can be associated with the friction-dependent rheology of granular systems and friction-induced granular segregations, and further generalized to applications with multiple species of particles in various natural and engineering mixtures. [ABSTRACT FROM AUTHOR]

The compaction of asphalt mixture is crucial to the performance of the pavement. However, the mix design (i.e., porosity, aggregate size distribution, binder content), which is based on compaction results, remains largely empirical. It is difficult to relate the aggregate size distribution and the asphalt binder properties to the compaction curve in both the field and laboratory compaction of asphalt mixtures. In this paper, the author proposes a simple mathematical model from the perspective of granular physics to predict the compaction of asphalt mixtures. In this model, the compaction process is divided into two mechanisms: (i) viscoplastic deformation of an ordered granular-fluid assembly, and (ii) the transition from an ordered system to a disordered system due to particle rearrangement. This model could take into account both the viscous properties of the asphalt binder and the grain size distributions of the aggregates, where the viscous deformation is calculated with a proposed governing equation and the particle rearrangement effect is solved using simple DEM simulations. This model is calibrated based on the Superpave gyratory compaction tests in the pavement lab, and the R-squares of model predictions are all above 0.95. The model results are compared with experimental data to show that it can provide good predictions for the experiments, suggesting its potential for enhancing the design of asphalt mixtures. [ABSTRACT FROM AUTHOR]

Objetivos: observar se o regime do erro médico previsto no Regulamento para o Tratamento de Acidentes Médicos perde ou não a sua utilidade prática a partir da entrada em vigor da Lei de Responsabilidade Civil da República Popular da China que prevê especificamente um regime da responsabilidade pelos danos causados por tratamentos médicos; segundo, ver se a constituição de acidente médico ainda é um pressuposto da responsabilidade civil médica prevista na lei citada; terceiro, ver se ambos os regimes, agora de diferentes diplomas legais, requerem os mesmos requisitos e se cada um destes tem o mesmo âmbito. Metodologia: executaram-se estudos bibliográficos e normativos que se relacionam com os dois regimes. Resultado e conclusão: o regime de acidente médico apenas perde a sua utilidade prática relativa à responsabilidade civil médica, mas ainda se mantém a sua utilidade prática de natureza administrativa. A constituição de acidente médico não é um pressuposto da responsabilidade civil médica prevista na nova lei. Os requisitos para ambos os regimes são o facto ilícito, negligência médica ou culpa, dano, nexo de causalidade e instituição médica e profissionais de saúde como sujeitos, mas alguns destes requisitos têm âmbito diferente, como nas partes relativas à ilicitue, à negligência médica ou culpa e ao dano.

MicroRNAs (miRNAs) are small non-coding RNAs that have been identified in animals, plants, and viruses. These small RNAs play important roles in post-transcriptional regulation of various cellular processes, including development, differentiation, and all aspects of cancer biology. Rapid-onset T-cell lymphoma of chickens, namely Marek’s disease (MD), induced by Gallid alphaherpesvirus 2 (GaHV2), could provide an ideal natural animal model for herpesvirus-related cancer research. GaHV2 encodes 26 mature miRNAs derived from 14 precursors assembled in three distinct gene clusters in the viral genome. One of the most highly expressed GaHV2 miRNAs, miR-M4-5p, shows high sequence similarity to the cellular miR-155 and the miR-K12-11 encoded by Kaposi’s sarcoma-associated herpesvirus, particularly in the miRNA “seed region.” As with miR-K12-11, miR-M4-5p shares a common set of host and viral target genes with miR-155, suggesting that they may target the same regulatory cellular networks; however, differences in regulatory function between miR-155 and miR-M4-5p may distinguish non-viral and viral mediated tumorigenesis. In this review, we focus on the functions of miR-M4-5p as the viral ortholog of miR-155 to explore how the virus mimics a host pathway to benefit the viral life cycle and trigger virus-induced tumorigenesis.

Marek’s disease virus (MDV), a lymphotropic α-herpesvirus associated with T-cell lymphomas in chickens, is an excellent model for herpesvirus biology and virus-induced oncogenesis. Marek’s disease (MD) is also one of the cancers against which a vaccine was first used. In the lymphomas and lymphoblastoid cell lines (LCLs) derived from them, MDV establishes latent infection with limited gene expression. Although LCLs are valuable for interrogating viral and host gene functions, molecular determinants associated with the maintenance of MDV latency and lytic switch remain largely unknown, mainly due to the lack of tools for in situ manipulation of the genomes in these cell lines. Here we describe the first application of CRISPR/Cas9 editing approach for precise editing of the viral gene phosphoprotein 38 (pp38), a biomarker for latent/lytic switch in MDV-transformed LCLs MDCC-MSB-1 (Marek’s disease cell line MSB-1) and MDCC-HP8. Contradictory to the previous reports suggesting that pp38 is involved in the maintenance of transformation of LCL MSB-1 cells, we show that pp38-deleted cells proliferated at a significant higher rate, suggesting that pp38 is dispensable for the transformed state of these cell lines. Application of CRISPR/Cas9-based gene editing of MDV-transformed cell lines in situ opens up further opportunities towards a better understanding of MDV pathogenesis and virus-host interactions.

We formulate a finite‐size scaling hypothesis to predict the global permeability of fracture networks. To validate the hypothesis, numerous discrete fracture networks are generated, and the permeability is numerically calculated. Results suggest that the dimensionless permeability, scaled by moments of local conductivity and fracture sizes and corrected by two stereological ratios, can capture variations in fracture attributes (orientations, sizes, and apertures). The universal form obtained in this study can also explain the contradictory observations where the permeability decreases or increases with the domain size of fracture networks. We show that how a clear transition point, where the permeability does not change with the domain size, is obtained from this universal form. This study provides a solid theoretical foundation to understand the connection between fracture attributes and field‐scale hydraulic properties. Plain Language Summary: Knowledge of the permeability of fractured rocks is critical for many geo‐engineering applications (such as deep geological disposal of radioactive wastes, mining and dam engineering) and environmental problems (such as ground water and pollutant transport). The influence of fracture attributes (e.g., orientations, sizes, and apertures) on hydraulic properties of rock masses is significant, but the task of correlating the permeability to individual fracture data is still a challenging one with remarkable knowledge gaps. In this letter, instead of using analytical solutions, we develop a finite‐size scaling (FSS) function, inspired by the percolation theory in statistic physics, to describe the permeability of fracture networks. The universality of the scaling function is validated by massive simulation data which cover a wide range of fracture attributes. Results demonstrate that, regardless of how domain sizes and fracture attributes change, the scaling for the permeability is of an invariant form, after considering the finite‐size effect. Our findings motivate further studies of applying the FSS function at the field scale and improve on the information that earth scientists can obtain from fracture statistics. Key Points: A finite‐size scaling hypothesis for the permeability of discrete fracture networks (DFNs) is formulated with several dimensionless quantitiesNumerous DFN representations and flow simulations are implemented to validate the scaling hypothesisThe scaling is found universal for different DFNs with an invariant form [ABSTRACT FROM AUTHOR]

Devido às profundas mudanças económicas e sociais na China a prestação de cuidados de saúde enfrenta sérias dificuldades para satisfazer as necessidades dos pacientes, conduzindo pois a frequentes conflitos entre pacientes e médicos e/ou estabelecimentos de cuidados de saúde. Uma vez que os tribunais comuns parecem incapazes de resolver os litígios médicos de uma forma considerada satisfatória para ambas as partes, o Governo Chinês criou os tribunais do acidente médico. Estes tribunais são caracterizados por duas notas principais: por um lado a sua composição, dado que, e ao contrário dos tribunais judiciais, são compostos por decisores com conhecimentos médicos; por outro lado, o facto de a actividade dos tribunais do acidente médico se restringir ao campo da responsabilidade médica, sendo que esta especialização lhes permite decidir os casos de uma forma mais expedita e supostamente mais justa. Contudo, e apesar do grande desenvolvimento representado pelos tribunais do acidente médico, eles representam apenas uma pequena peça da grande reforma exigida pelos conflitos médicos que têm lugar por toda a China.

Very few studies have emphasized the effects of high-pressure sintering on snow density evolution, even though snow as a type of engineering material is widely used in construction engineering in cold regions for snow pavement, snow runway and polar infrastructure. This study presents new experimental results of snow densification under high pressures of up to 100 MPa for a temperature range from −3.5 to −17.3°C and uniaxial compression at the temperature of −10°C and constant strain rates from 5 × 10−4 to 10−1 s−1. Results reveal that density evolution of snow to ice under high-pressure sintering can be achieved in a wide temperature range within a duration as short as 5 min. The compressive strength of snow-sintered ice was ~1.2–2.2 times as large as that of water-frozen ice reported by previous work. The orthogonal experiment showed that pressure is a more significant factor affecting the final density in comparison with sintering temperature and time. The increased rates of ice fabrication, low limitations on temperature and reliable sintered snow strength indicate that snow-ice engineering, such as airport construction in Greenland and Antarctica, can be improved by high-pressure sintering of snow to overcome the harsh environment. [ABSTRACT FROM AUTHOR]