LORF9 of Marek's disease virus is involved in the early cytolytic replication of B lymphocytes and can act as a target for gene deletion vaccine development.

IMPORTANCE: Marek's disease virus (MDV) is a highly infectious and oncogenic virus that can induce severe T cell lymphomas in chickens. MDV encodes more than 100 genes, most of which have unknown functions. This work indicated that the LORF9 gene is necessary for MDV early cytolytic replication in B lymphocytes. In addition, we have found that the LORF9 deletion mutant has a comparative immunological protective effect with CVI988/Rispens vaccine strain against very virulent MDV challenge. This is a significant discovery that LORF9 can be exploited as a possible target for the development of an MDV gene deletion vaccine.

Infection phase-dependent dynamics of the viral and host N6-methyladenosine epitranscriptome in the lifecycle of an oncogenic virus in vivo.

Dynamic alteration of the epitranscriptome exerts regulatory effects on the lifecycle of oncogenic viruses in vitro. However, little is known about these effects in vivo because of the general lack of suitable animal infection models of these viruses. Using a model of rapid-onset Marek's disease lymphoma in chickens, we investigated changes in viral and host messenger RNA (mRNA) N6-methyladenosine (m6 A) modification during Marek's disease virus (MDV) infection in vivo. We found that the expression of major epitranscriptomic proteins varies among viral infection phases, reprogramming both the viral and the host epitranscriptomes. Specifically, the methyltransferase-like 3 (METTL3)/14 complex was suppressed during the lytic and reactivation phases of the MDV lifecycle, whereas its expression was increased during the latent phase and in MDV-induced tumors. METTL3/14 overexpression inhibits, whereas METTL3/14 knockdown enhances, MDV gene expression and replication. These findings reveal the dynamic features of the mRNA m6 A modification program during viral replication in vivo, especially in relation to key pathways involved in tumorigenesis.

Research Progress of Long Non-coding RNAs in Spinal Cord Injury.

Spinal cord injury (SCI) can result in a partial or complete loss of motor and sensory function below the injured segment, which has a significant impact on patients' quality of life and places a significant social burden on them. Long non-coding RNA (LncRNA) is a 200-1000 bp non-coding RNA that has been shown to have a key regulatory role in the progression of a variety of neurological illnesses. Many studies have demonstrated that differentially expressed LncRNAs following spinal cord injury can participate in inflammatory damage, apoptosis, and nerve healing by functioning as competitive endogenous RNA (ceRNA); at the same time, it has a significant regulatory effect on sequelae such neuropathic pain. As a result, we believe that LncRNAs could be useful as a molecular regulatory target in the diagnosis, treatment, and prognosis of spinal cord injury.

Effect of Asymmetry on the Contact Time of Droplet Impact on Superhydrophobic Cylindrical Surfaces.

Reducing the contact time during the droplet impact on the surface is crucial for anti-icing, self-cleaning, and heat transfer optimization applications. This study aims to minimize the contact time by modifying the surface curvature to create an asymmetric impact process. Our experiments showed that the increase in Weber numbers (We) and the decrease in the ratio of surface diameter to droplet diameter (D*) intensify the asymmetry of the impact process, yielding four distinct rebound modes. Low asymmetry observes the liquid retract toward the central point (Rebound Modes 1 and 2), whereas high asymmetry yields a wing-like rebound (Rebound Modes 3 and 4). In Rebound Mode 1, increased asymmetry would lead to more extended contact due to the prolonged waiting period. Conversely, the reduction in contact time in Rebound Mode 2 occurs due to increased asymmetry with no waiting period. For Rebound Modes 3 and 4, the retraction time could be divided into three stages, generated by two liquid detachment modes from the surface. Analysis reveals that an increased asymmetry would reduce the retraction time during the first stage but prolong it during the third stage, with no significant effects on the second. Four correlations, each pertaining to a distinct impact mode, are proposed based on these analyses to describe the contact time concerning We and D* for droplets impacting a superhydrophobic cylindrical surface.

Monitoring nature's calendar from space: Emerging topics in land surface phenology and associated opportunities for science applications.

Vegetation phenology has been viewed as the nature's calendar and an integrative indicator of plant-climate interactions. The correct representation of vegetation phenology is important for models to accurately simulate the exchange of carbon, water, and energy between the vegetated land surface and the atmosphere. Remote sensing has advanced the monitoring of vegetation phenology by providing spatially and temporally continuous data that together with conventional ground observations offers a unique contribution to our knowledge about the environmental impact on ecosystems as well as the ecological adaptations and feedback to global climate change. Land surface phenology (LSP) is defined as the use of satellites to monitor seasonal dynamics in vegetated land surfaces and to estimate phenological transition dates. LSP, as an interdisciplinary subject among remote sensing, ecology, and biometeorology, has undergone rapid development over the past few decades. Recent advances in sensor technologies, as well as data fusion techniques, have enabled novel phenology retrieval algorithms that refine phenology details at even higher spatiotemporal resolutions, providing new insights into ecosystem dynamics. As such, here we summarize the recent advances in LSP and the associated opportunities for science applications. We focus on the remaining challenges, promising techniques, and emerging topics that together we believe will truly form the very frontier of the global LSP research field.

Quantifying the influences of climate change and human activities on the grassland in the Southwest Transboundary Basin, China.

The Southwest Transboundary Basin (STB), which contains a part of the Tibetan Plateau and Southwest China, covers a large area of grassland that has been experiencing significant climate change (Cc) and human activities (Ha). However, variabilities of the grasslands and respective contributions of the drivers have not been fully understood. In this study, we quantitatively investigated the influences of Cc and Ha on the grasslands from 1982 to 2015 across the STB, using residual analysis, trend analysis, and partial correlation analysis. Particularly, vapor pressure deficit (VPD), which is increasingly paid attention to in current studies, was included in our analysis as a critical driver of alpine grasslands. Additionally, a new method, pure climate-induced grassland pixel, which combined the coefficient of variation, was proposed to significantly improve the pixel purity with higher coefficient correlation and lower error and bias. Results show that changes in the climate play an important role in the grassland coverage of STB during 1982-1999 and 2000-2015, while Ha only had a relatively weak negative impact on grassland during 2000-2015, mainly in the Lancang River Basin. Specifically, the grasslands showed an increasing trend in 1982-1999, dominated by climate change with a contribution rate of 80.87%. Temperature had the greatest contribution, followed by VPD and precipitation in this period. On the contrary, the grasslands showed a decreasing trend in 2000-2015, dominated by both Cc and Ha with the contributions of 53.16% and 46.84%, respectively. The findings of this study provide a valuable reference for future ecological restoration in other similar alpine regions.

The insufficiency of ATG4A in macroautophagy.

During autophagy, LC3 and GABARAP proteins become covalently attached to phosphatidylethanolamine on the growing autophagosome. This attachment is also reversible. Deconjugation (or delipidation) involves the proteolytic cleavage of an isopeptide bond between LC3 or GABARAP and the phosphatidylethanolamine headgroup. This cleavage is carried about by the ATG4 family of proteases (ATG4A, B, C, and D). Many studies have established that ATG4B is the most active of these proteases and is sufficient for autophagy progression in simple cells. Here we examined the second most active protease, ATG4A, to map out key regulatory motifs on the protein and to establish its activity in cells. We utilized fully in vitro reconstitution systems in which we controlled the attachment of LC3/GABARAP members and discovered a role for a C-terminal LC3-interacting region on ATG4A in regulating its access to LC3/GABARAP. We then used a gene-edited cell line in which all four ATG4 proteases have been knocked out to establish that ATG4A is insufficient to support autophagy and is unable to support GABARAP proteins removal from the membrane. As a result, GABARAP proteins accumulate on membranes other than mature autophagosomes. These results suggest that to support efficient production and consumption of autophagosomes, additional factors are essential including possibly ATG4B itself or one of its proteolytic products in the LC3 family.

Rebound Behaviors of Multiple Droplets Simultaneously Impacting a Superhydrophobic Surface.

The rebound behaviors of multiple droplets simultaneously impacting a superhydrophobic surface were investigated via lattice Boltzmann method (LBM) simulations. Three rebound regions were identified, i.e., an edge-dominating region, a center-dominating region, and an independent rebound region. The occurrence of the rebound regions strongly depends on the droplet spacing and the associated Weber and Reynolds numbers. Three new rebound morphologies, i.e., a pin-shaped morphology, a downward comb-shaped morphology, and an upward comb-shaped morphology, were presented. Intriguingly, in the edge-dominating region, the central droplets experience a secondary wetting process to significantly prolong the contact time. However, in the center-dominating region, the contact time is dramatically shortened because of the strong interactions generated by the central droplets and the central ridges. These findings provide useful information for practical applications such as self-cleaning, anticorrosion, anti-icing, and so forth.

Comparative transcriptome analysis of Chinese grass shrimp (Palaemonetes sinensis) hepatopancreas under ectoparasitic isopod (Tachaea chinensis) infection.

Tachaea chinensis, a parasitic isopod, negatively affects the production of several commercially important shrimp species. To better understand the interaction between shrimp immunity and isopod infection, we performed a transcriptome analysis of the hepatopancreas of Palaemonetes sinensis challenged with T. chinensis. After assembly and annotation, 75,980 high-quality unigenes were obtained using RNA-seq data. Differential gene expression analysis revealed 896 significantly differently expressed genes (DEGs) after infection, with 452 and 444 upregulated and downregulated genes, respectively. Specifically, expression levels of genes involved in detoxification, such as the interferon regulatory factor, venom carboxylesterase-6, serine proteinase inhibitor, and cytochrome P450, were upregulated. Furthermore, expression levels of genes corresponding to retinol dehydrogenase, triosephosphate isomerase, variant ionotropic glutamate receptor, and phosphoenolpyruvate carboxykinase were significantly upregulated after isopod parasitization, indicating that the shrimp's visual system was influenced by isopod parasitization. Moreover, quantitative real-time PCR of 10 DEGs helped validate the RNA-seq findings. These results provide a valuable basis for future studies on the elucidation of immune responses of P. sinensis to T. chinensis infection.

Life cycle of the ectoparasite Tachaea chinensis (Isopoda: Corallanidae) on the freshwater shrimp Palaemonetes sinensis (Decapoda: Palaemonidae).

In this study, we describe in detail the life cycle of Tachaea chinensis (Isopoda: Corallanidae), a branchial ectoparasitic isopod that infests the freshwater shrimp Palaemonetes sinensis in China. We obtained 14 ovigerous T. chinensis females (8.22-11.92 mm in length) and observed the development of embryos through 5 sequential ontogenetic stages within the brood pouches (marsupium) of these females. The number of eggs or mancae (post-larval juveniles) held in the female marsupium ranged from 31 to 86, with a mean ± SD of 61.25 ± 16.16 eggs. Female T. chinensis were semelparous, i.e. individuals died following the release of mancae from the marsupium. Released mancae were non-planktonic and immediately infective to host shrimps. However, only a few mancae successfully established contact with a host, and it is thus assumed that the remainder were predated by shrimp. Attached T. chinensis fed on the host hemolymph, and subsequent to host death, these isopods typically searched for a new host. We also found that T. chinensis exhibits a host preference: most mancae attached to P. sinensis rather than to Neocaridina sp. or Macrobrachium nipponense. This study provides valuable empirical data that will support future research on the prevention and control of parasitic isopod infections.

Concurrent and lagged effects of spring greening on seasonal carbon gain and water loss across the Northern Hemisphere.

Spring greening has been widely observed across the Northern Hemisphere (NH) using a remotely sensed vegetation index (e.g., the normalized difference vegetation index, NDVI). However, there is still a debate on the ecological effects of spring greening on seasonal carbon and water budgets. This study jointly investigated the concurrent and lagged effects of spring greening on carbon gain (gross primary productivity, GPP) and water loss (evapotranspiration, ET) in the summer-active ecosystems at mid and high latitudes of NH using remote sensing and multimodel ensemble data during 1982-2013. The results showed that the collective promotion of spring greening to concurrent GPP and ET is widespread despite variations in magnitude and significance. Both beneficial and adverse lagged effects of spring greening on summer GPP commonly appear with an obvious spatial heterogeneity and difference among climate-plant types. However, the expected significant suppression of spring greening to summer GPP was rarely observed even in the areas where spring ET was significantly promoted by spring greening. Nevertheless, when drought was taken into account, the response patterns of spring water use to spring greening varied to some extent, and the adverse lagged effect of spring greening to summer GPP appeared or strengthened in some regions, especially during the years with dry summer. Given the predicted warming of the climate and more frequent climatic extremes, the adverse effect of spring greening should be given more attention.

α-Mangostin promotes apoptosis of human rheumatoid arthritis fibroblast-like synoviocytes by reactive oxygen species-dependent activation of ERK1/2 mitogen-activated protein kinase.

α-Mangostin (α-M) is a commonly used traditional medicine with various biological and pharmacological activities. Our study aimed to explore the effects and mechanism of α-M in regulating apoptosis of rheumatoid arthritis fibroblast-like synoviocytes (RA-FLS). α-M of 10 to 100 µM was used to treat RA-FLS for 24 hours, followed by measuring cell viability and apoptosis. The involvement of reactive oxygen species (ROS) and mitogen-activated protein kinases was detected. Treatment of α-M promoted apoptosis and reduced viability of RA-FLS in a dose-dependent manner. The mitochondrial membrane potential in RA-FLS was remarkably reduced by α-M treatment, accompanied by the cytochrome c accumulation in the cytosol and increased activities of caspase-3 and caspase-9. Moreover, we found that α-M treatment promoted ROS production and extracellular signal-regulated kinase 1/2 (ERK1/2) phosphorylation. The proapoptotic activity of α-M in RA-FLS was markedly reversed by the co-induction with the ERK1/2 inhibitor LY3214996 or ROS scavenger N-acetyl-l-cysteine. In conclusion, our studies found that α-M had remarkable proapoptotic activities in RA-FLS, which is regulated by the induction of ROS accumulation and ERK1/2 phosphorylation. α-M may thus have potential therapeutic effects for rheumatoid arthritis.

MicroRNA-185 inhibits the growth and proliferation of osteoblasts in fracture healing by targeting PTH gene through down-regulating Wnt/ß -catenin axis: In an animal experiment.

Fracture healing is a repair process of a mechanical discontinuity loss of force transmission, and pathological mobility of bone. Increasing evidence suggests that microRNA (miRNA) could regulate chondrocyte, osteoblast, and osteoclast differentiation and function, indicating miRNA as key regulators of bone formation, resorption, remodeling, and repair. Hence, during this study, we established a right femur fracture mouse model to explore the effect microRNA-185 (miR-185) has on osteoblasts in mice during fracture healing and its underlying mechanism. After successfully model establishment, osteoblasts were extracted and treated with a series of mimics or inhibitors of miR-185, or siRNA against PTH. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blot analysis were performed to determine the levels of miR-185, PTH, ß-catenin and Wnt5b. Cell viability, cycle distribution and apoptosis were detected by means of MTT and flow cytometry assays. Dual luciferase reporter gene assay verified that PTH is a target gene of miR-185. Osteoblasts transfected with miR-185 mimics or siRNA against PTH presented with decreased levels of PTH, ß-catenin and Wnt5b which indicated that miR-185 blocks the Wnt/ß -catenin axis by inhibiting PTH. Moreover, miR-185 inhibitors promoted the osteoblast viability and reduced apoptosis with more cells arrested at the G1 stage. MiR-185 mimics were observed to have inhibitory effects on osteoblasts as opposed to those induced by miR-185 inhibitors. Above key results indicated that suppression of miR-185 targeting PTH could promote osteoblast growth and proliferation in mice during fracture healing through activating Wnt/ß -catenin axis.

Ammonia Emissions May Be Substantially Underestimated in China.

China is a global hotspot of atmospheric ammonia (NH3) emissions and, as a consequence, very high nitrogen (N) deposition levels are documented. However, previous estimates of total NH3 emissions in China were much lower than inference from observed deposition values would suggest, highlighting the need for further investigation. Here, we reevaluated NH3 emissions based on a mass balance approach, validated by N deposition monitoring and satellite observations, for China for the period of 2000 to 2015. Total NH3 emissions in China increased from 12.1 ± 0.8 Tg N yr-1 in 2000 to 15.6 ± 0.9 Tg N yr-1 in 2015 at an annual rate of 1.9%, which is approximately 40% higher than existing studies suggested. This difference is mainly due to more emission sources now having been included and NH3 emission rates from mineral fertilizer application and livestock having been underestimated previously. Our estimated NH3 emission levels are consistent with the measured deposition of NHx (including NH4+ and NH3) on land (11-14 Tg N yr-1) and the substantial increases in NH3 concentrations observed by satellite measurements over China. These findings substantially improve our understanding on NH3 emissions, implying that future air pollution control strategies have to consider the potentials of reducing NH3 emission in China.

Survival and disease-free benefits with mastectomy versus breast conservation therapy for early breast cancer: a meta-analysis.

The objective of the present meta-analysis was to estimate the magnitude of survival and disease-free benefits from mastectomy compared with breast conservation therapy (BCT) in patients with early breast cancer. We searched PubMed, Embase, the Cochrane Library, Web of Science, and Chinese biomedical literature database from their inception to May 2015. All the data were independently extracted from the publications by two reviewers. Results regarding the overall survival (OS) and disease-free survival (DFS) in the meta-analysis were expressed as hazard ratios (HRs) with 95 % confidence intervals (CIs). Nine randomized control trials were eligible for final meta-analysis. Meta-analysis showed that mastectomy provided significant benefit in OS compared with BCT (HR 1.09, 95 % CI 1.01-1.19; P = 0.03). Sensitivity analysis gives similar OS estimates (HR 1.12, 95 % CI 1.01-1.25). In the subgroup analysis of patients according to tumor size, the pooled HRs for OS indicated that there is a borderline statistical difference between two arms in the subgroup with tumor size ranging between ≥2 cm and <5 cm (HR 1.09, 95 % CI 1.00-1.19), but subgroup analysis of tumor size <2 cm showed no statistically significant difference in OS (HR 1.08, 95 % CI 0.88-1.33) when comparing the BCT arm with the mastectomy arm. There was no significant difference in DFS between BCT and mastectomy groups (HR 1.08, 95 % CI 0.99-1.18; P = 0.08). Sensitivity analysis also gives similar DFS estimates (HR 1.11, 95 % CI 0.96-1.27). Subgroup analysis indicated that the pooled HRs for DFS did not favor mastectomy arm or BCT arm either in the subgroup with tumor size <2 cm (HR 1.09, 95 % CI 0.78-1.52) or in the subgroup with tumor size ranging between ≥2 cm and <5 cm (HR 1.08, 95 % CI 0.99-1.18) according to tumor size. Five-year OS decreased from 70 to 68 % with BCT. The present meta-analysis indicated that mastectomy might provide slight OS benefit compared with BCT in early breast cancer patients with larger tumor size, but the absolute survival gain is small.

Development of an isothermoal amplification-based assay for rapid visual detection of an Orf virus.

BACKGROUND: Orf virus (ORFV) is the causative agent of a severe infectious skin disease (also known as contagious ecthyma) in goats, sheep and other small ruminants. Importantly, ORFV also infect humans which causes a public health concern in the context of changing environment and increase in human populations. The rapid detection is critical in effective control of the disease and urgently needed. RESULTS: A novel "point of care" molecular amplification assay for rapid visual detection of ORFV was developed based on isothermoal recombinase polymerase amplification (RPA) technology in combination with a simpler lateral flow immunoassay strip (ORFV RPA- LFD assay). The developed ORFV RPA- LFD assay was able to detect ORFV in less than 25 min. This assay was highly sensitive, with detection limit of as low as 80 copies/reaction, and highly specific, with no cross-reactions with capripox virus, foot-and-mouth disease virus and peste des petits ruminants virus. Furthermore, the ORFV RPA- LFD assay has good correlation with qPCR assay for detection of ORFV present in clinical samples. CONCLUSIONS: The developed ORFV RPA-LFD assay was a sensitive and specific method for rapid detection of ORFV, and has great potential as an onsite molecular diagnostic tool in control of Orf.

Comprehensive analysis of the immune implication of EPHX4 gene in laryngeal squamous cell carcinoma.

OBJECTIVES: The role of Epoxide Hydrolase-4 (EPHX4), a member of epoxide hydrolase family, has not been investigated in cancer. The purpose of this article is to explore the application value of EPHX4 in laryngeal cancer and its relationship with immune infiltration. METHODS: We observed that EPHX4 expression and its survival assays in laryngeal cancer specimens based on The Cancer Genome Atlas (TCGA) cohorts. We also analyzed the correlation between immune cell infiltration levels and EPHX4 gene copy number in laryngeal cancer. Finally, we conducted in vitro assay to evaluate the functions of EPHX4 in laryngeal cancer cell line. RESULTS: EPHX4 is highly expressed in laryngeal cancer specimens and has a poor prognosis. EPHX4 related immune cell analysis showed that it participated in NK Natural killer cell mediated cytotoxicity. Finally, Cell experiments indicate that EPHX4 could promote laryngeal cancer cell line proliferation, colony formation and invasion. CONCLUSIONS: Our research results suggest that EPHX4 may be a potential immunotherapy target for laryngeal cancer. The nominated immune signature is a helpful and promising prognostic indicator in laryngeal cancer. LEVELS OF EVIDENCE: Level 3.

Assessing the structural and foaming property changes in egg yolk proteins due to malondialdehyde: Experimental and molecular docking studies.

This study evaluated the effects of varying levels of malondialdehyde (MDA) on the structural and foaming properties of the egg yolk proteins (EYPs), and the interaction between them was explored by molecular docking. The results showed that oxidative modification due to MDA increased the carbonyl content of EYPs by 4.49 times. Simultaneously, the total sulfhydryl content was reduced by 21.47%, and the solubility of EYPs was significantly decreased (p < 0.05). Continuous oxidation disorders the previously ordered structure of EYPs. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicated that some proteins underwent crosslinking and aggregation with increased MDA oxidation, aligning with changes in particle size and zeta-potential. Moderate oxidation (<1 mmol/L) enhanced the foaming capacity and foam stability of EYPs. Additionally, molecular docking results uncovered favorable interactions between MDA and specific EYPs, primarily through hydrogen bonding. This research offers valuable insights into managing the functional and quality changes of yolk products during processing.

Evaluation of systemic medications associated with diabetic retinopathy: a nested case-control study from the UK Biobank.

AIMS: This study aims to investigate the associations between commonly used systemic medications and diabetic retinopathy (DR). METHODS: Individuals with linked primary care prescription data from the UK Biobank were included. Cases were defined as individuals with a Hospital Episode Statistics-coded or primary care recorded diagnosis of DR or self-reported DR. Controls were matched for age, sex, glycosylated haemoglobin, duration of diabetes mellitus (DM), hypertension status and cardiovascular disease status. ORs and 95% CIs were calculated using conditional univariate and multivariable logistic regression models. RESULTS: A total of 3377 case subjects with DR were included in the study and matched with 3377 control subjects. In multivariable logistic regression, increased odds of incident DR were observed for exposure to short-acting insulins (OR 1.63; 95% CI 1.22 to 2.18), medium-acting insulins (OR 2.10; 95% CI 1.60 to 2.75), sulfonylureas (OR 1.30; 95% CI 1.16 to 1.46). Instead, the use of fibrates (OR 0.71; 95% CI 0.53 to 0.94) and Cox-2 inhibitors (OR 0.68; 95% CI 0.58 to 0.79) was associated with decreased odds of incident DR. Dose-response relationships were observed for all five drug categories (all p<0.05). CONCLUSIONS: This study comprehensively investigated the associations between systemic medication use and DR and found significant associations between the use of short-acting insulins, medium-acting insulins and sulfonylureas with increased odds of incident DR. In contrast, fibrates and Cox-2 inhibitors were associated with decreased odds of incident DR. These findings may provide valuable insights into DM medication management and serve as a reference for the prevention of DR in patients with DM.