Percutaneous versus traditional open approaches for the treatment of thoracolumbar fractures in patients without neurologic deficits: a meta-analysis of 35 cohort studies.

At present, percutaneous surgery is widely used to treat thoracolumbar fractures. However, the actual safety, feasibility, and effectiveness of percutaneous surgery are not clear. Through systematic review and meta-analysis, we compared the efficacies of percutaneous pedicle screw fixation and open pedicle screw fixation in the treatment of thoracolumbar fractures without nerve root symptoms. We systematically searched the PubMed, Embase, and Cochrane libraries for articles published on or before June 2023. All results were evaluated by standard methods recommended for meta-analysis, continuous data were expressed by standard mean differences (SMDs), and binary variables were analyzed by odds ratios (ORs) and 95% confidence intervals (95% CIs). We also explored the main sources of heterogeneity and the stability of the results through sensitivity analysis, Begg's funnel plots, and Egger's test. Thirty-five cohort studies with a total of 3039 patients were included. The study found that patients who undergo percutaneous approaches have less intraoperative blood loss (IBL), shorter length of hospital stay (LOS), shorter operation time, and shorter incision. Moreover, percutaneous approaches had more advantages in terms of visual analog scale (VAS) scores, Oswestry Disability Index (ODI) scores, and infection rates. However, there was no significant difference in anterior vertebral body height (AVB), Cobb angle (CA), or screw errors between the two groups. In the long run, the clinical and surgical results of the percutaneous approach are better than those of the open approach, but the radiological results of both operations do not seem to show an advantage for any specific approach. Because of publication bias and heterogeneity, our findings must be interpreted with caution. However, this paper will provide some support for clinicians to choose suitable surgical methods for the treatment of thoracolumbar fractures.

Lappaol F regulates the cell cycle by activating CDKN1C/p57 in human colorectal cancer cells.

CONTEXT: Lappaol F (LAF), a natural lignan from Arctium lappa Linné (Asteraceae), inhibits tumor cell growth in vitro and in vivo. The underlying mechanism involves the suppression of the Yes-associated protein. However, the specific role of LAF in cell cycle regulation remains unknown. OBJECTIVE: This study determined the molecular mechanism by which LAF regulates cell cycle progression. MATERIALS AND METHODS: Various colon cancer cell lines (SW480, HCT15, and HCT116) were treated with LAF (25, 50, and 75 µmol/L) for 48 h. The effects of LAF on cell proliferation and cell cycle were determined using sulforhodamine B and flow cytometry assays. Differentially expressed proteins (DEPs) were identified using quantitative proteomics. Bioinformatic analysis of DEPs was conducted via Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses. Expression levels of DEPs in the cell cycle pathway were analyzed using RT-qPCR and western blotting. RESULTS: LAF suppressed the proliferation of SW480, HCT15, and HCT116 cells (IC50 47.1, 51.4, and 32.8 µmol/L, respectively) and induced cell cycle arrest at the S phase. A total of 6331 proteins were identified and quantified, of which 127 were differentially expressed between the LAF-treated and untreated groups. GO and KEGG enrichment analyses revealed that DEPs mainly participated in the cell cycle. CDKN1C/p57 showed the most significant differential expression, with the highest fold-change (3.155-fold). Knockdown of CDKN1C/p57 attenuated the S phase cell cycle arrest and proliferation inhibition induced by LAF. CONCLUSION: LAF exerts antitumor effects via S phase arrest by activating CDKN1C/p57 in colorectal cancer cells.

Formicincola oecophyllae gen. nov. sp. nov., a novel member of the family Acetobacteraceae isolated from the weaver ant Oecophylla smaragdina.

In this study, a novel bacterium designated F3b2T was isolated from the gut sample of weaver ant Oecophylla smaragdina and characterised. Strain F3b2T was a Gram-negative, aerobic, non-motile, ovoid-shaped bacterium and grows optimally at 28-30 °C. Its major respiratory quinone is ubiquinone 10 (Q-10) and the major fatty acids are C18:1 ω7c, C19:0 cyclo ω8c and C16:0, representing 85% of the total fatty acids. The 16S rRNA gene sequence of strain F3b2T was highest in similarity to that of Oecophyllibacter saccharovorans DSM106907T and Swingsia samuieinsis NBRC 107927T at 94.35% and 91.96%, respectively. A 16S rRNA gene-based phylogenetic analysis and a core genes-based phylogenomic analysis placed strain F3b2T in a distinct lineage in the family Acetobacteraceae. The phylogenetic placement was supported by lower than species delineation threshold average nucleotide identity (ANI) (≤ 70.2%), in silico DNA-DNA hybridization (DDH) (≤ 39.5%) and average amino acid identity (AAI) (≤ 63.5%) values between strain F3b2T and closest neighbours. These overall genome relatedness indices also supported the assignment of strain F3b2T to a novel genus within Acetobacteraceae. The genome of strain F3b2T was 1.96 Mb with 60.4% G + C DNA content. Based on these results, strain F3b2T represented a novel taxon of Acetobacteraceae, for which we proposed the name Formicincola oecophyllae gen. nov. sp. nov., and strain F3b2T (= LMG 30590T = DSM 106908T = NBRC 113640T = KCTC 62951T) as the type strain.

Effectiveness of Public Education Campaigns for Stroke Symptom Recognition and Response in Non-Elderly Adults: A Systematic Review and Meta-Analysis.

OBJECTIVES: The incidence of stroke is on the rise among younger adults. Stroke educational campaigns are often used to raise public knowledge of stroke warning signs and response actions, but their effectiveness in non-elderly adults is unclear. This study investigated the effectiveness of stroke campaigns in improving stroke symptom recognition and intention to call emergency medical services in adults aged < 65 years, and described the characteristics of public stroke education in this demographic group. MATERIALS AND METHODS: Seven databases (Cochrane Library, CINAHL, Embase, Medline, PsycINFO, Scopus, and ProQuest Dissertations and Theses) were searched from inception to 24 January 2021. A meta-analysis was conducted to synthesize the effects of stroke education campaigns on stroke symptom recognition and intention to call emergency medical services in adults < 65 years. A narrative synthesis approach was used to explore the types of educational content and campaign best suited to this demographic group. RESULTS: Thirteen studies from 2001 to 2019 were included in this review. Out of the thirteen studies, five were included in the meta-analysis to synthesize the effects of stroke education campaigns on symptom recognition and four were included for synthesizing the effects on intention to call emergency medical services. The estimated pooled risk ratio for post-campaign compared with pre-campaign in stroke symptom recognition and intention to call emergency medical services was 1.20 (95% CI: 1.07-1.36, p = 0.002) and 1.19 (95% CI: 1.11-1.28, p < 0.00001), respectively. Usage of acronyms in broadcast and digital media campaigns appears promising in enhancing recognition of and responsiveness to stroke symptoms. CONCLUSIONS: Public stroke education campaigns were found to have a significant impact on stroke symptom recognition and intention to call emergency medical services. The evidence generated from this review could be applied to inform future campaigns targeted at younger adults.

Lappaol F, an anticancer agent, inhibits YAP via transcriptional and post-translational regulation.

CONTEXT: Lappaol F (LAF), a natural lignan from Arctium lappa Linné (Asteraceae), inhibits tumour cell growth by inducing cell cycle arrest. However, its underlying anticancer mechanism remains unclear. OBJECTIVE: The effects of LAF on the Hippo-Yes-associated protein (YAP) signalling pathway, which plays an important role in cancer progression, were explored in this study. MATERIALS AND METHODS: Cervical (HeLa), colorectal (SW480), breast (MDA-MB-231) and prostate (PC3) cancer cell lines were treated with LAF at different concentrations and different durations. BALB/c nude mice bearing colon xenografts were intravenously injected with vehicle, LAF (10 or 20 mg/kg) or paclitaxel (10 mg/kg) for 15 days. The expression and nuclear localisation of YAP were analysed using transcriptome sequencing, quantitative PCR, western blotting and immunofluorescence. RESULTS: LAF suppressed the proliferation of HeLa, MDA-MB-231, SW480 and PC3 cells (IC50 values of 41.5, 26.0, 45.3 and 42.9 µmol/L, respectively, at 72 h), and this was accompanied by significant downregulation in the expression of YAP and its downstream target genes at both the mRNA and protein levels. The expression of 14-3-3σ, a protein that causes YAP cytoplasmic retention and degradation, was remarkably increased, resulting in a decrease in YAP nuclear localisation. Knockdown of 14-3-3σ with small interfering RNA partially blocked LAF-induced YAP inhibition and anti-proliferation effects. In colon xenografts, treatment with LAF led to reduced YAP expression, increased tumour cell apoptosis and tumour growth inhibition. CONCLUSION: LAF was shown to be an inhibitor of YAP. It exerts anticancer activity by inhibiting YAP at the transcriptional and post-translational levels.

Quorum sensing activity of Hafnia alvei isolated from packed food.

Quorum sensing (QS) is a mechanism adopted by bacteria to regulate expression of genes according to population density. N-acylhomoserine lactones (AHLs) are a type of QS signalling molecules commonly found in Gram-negative bacteria which have been reported to play a role in microbial spoilage of foods and pathogenesis. In this study, we isolated an AHL-producing Hafnia alvei strain (FB1) from spherical fish pastes. Analysis via high resolution triple quadrupole liquid chromatography/mass spectrometry (LC/MS) on extracts from the spent supernatant of H. alvei FB1 revealed the existence of two short chain AHLs: N-(3-oxohexanoyl) homoserine lactone (3-oxo-C6-HSL) and N-(3-oxo- octanoyl) homoserine lactone (3-oxo-C8-HSL). To our knowledge, this is the first report of the production of AHLs, especially 3-oxo-C8-HSL, by H. alvei.

Carbon emission prediction of construction industry in Sichuan Province based on the GA-BP model.

The reduction of the carbon emissions of construction industry is urgent. Therefore, it is essential to accurately predict the carbon emissions of the provincial construction industry, which can support differentiation emission reduction policies in China. This paper proposes a carbon emission prediction model that optimizes the backpropagation (BP) neural network by genetic algorithm (GA) to predict carbon emission of construction industry, or "GA-BP". To begin with, the carbon emissions of construction industry in Sichuan Province from 2000 to 2020 are calculated by the emission factor method. Further, the electricity correction factor is introduced to eliminate the regional difference in electricity carbon emission coefficient. Finally, four factors are selected by the grey correlation analysis method to predict the carbon emission of construction industry in Sichuan Province from 2021 to 2025. The results show that the carbon emissions of construction industry in Sichuan Province have been trending up in the past two decades, with an average increase rate of 10.51%. The GA-BP model is a high-precision prediction model to predict carbon emissions of construction industry. The mean absolute percentage error (MAPE) of the model is only 6.303%, and its coefficient of determination is 0.853. Moreover, the carbon emissions of construction industry in Sichuan Province will reach 8891.97 million tons of CO2 in 2025. The GA-BP model can effectively predict the future carbon emissions of construction industry in Sichuan Province, which provides a new idea for the green and sustainable development of construction industry in Sichuan Province.

Sewage sludge-derived nutrients and biostimulants stimulate rice leaf photosynthesis and root metabolism to enhance carbohydrate, nitrogen and antioxidants accumulation.

The production of high quality liquid nitrogen fertilizer with both nutrient comprehensive and biostimulant properties by alkaline thermal hydrolysis of sewage sludge has shown great potential in agricultural production. However, little is known about the effects of sewage sludge-derived nutrients, and biostimulants (SS-NB) on leaf photosynthesis and root growth in rice. Phenotypic, metabolic and microbial analyses were used to reveal the mechanism of SS-NB on rice. Compared to NF treatment, phenotypic parameters (fresh/dry weight, soluble sugar, amino acid, protein) were increased by SS-NB in rice. SS-NB can enhance the photosynthesis of rice leaves by improving the photoconversion efficiency, chlorophyll content, ATP synthase activity, Rubisco and NADPH production. Meanwhile, SS-NB also increased antioxidant capacity (SOD, POD, CAT and proline) in rice leaf and root tissues. Metabolomics revealed that SS-NB application increased the expression levels of metabolites in root and leaf tissues, including carbohydrate, nitrogen and sulfur metabolism, amino acid metabolism, antioxidants, and phytohormone. Most importantly, the regulation of metabolites in rice root tissues is more sensitive than in leaf tissues, especially to the higher levels of antioxidants and phytohormones (IAA and GA) in rice root tissues. Furthermore, SS-NB increased the abundance of photosynthetic autotrophic, organic acids-degrading and denitrifying functional bacteria in rice roots and recruited plant growth-promoting bacteria (Azospirillum and norank_f_JG30-KF-CM45), while the NF treatment group resulted in an imbalance of the microbial community, leading to the dominance of pathogenic bacteria. The results showed that SS-NB had great application potential in crop growth and stress resistance improvement.

STING signalling compensates for low tumour mutation burden to drive anti-tumour immunity.

BACKGROUND: While mutation-derived neoantigens are well recognized in generating anti-tumour T cell response, increasing evidences highlight the complex association between tumour mutation burden (TMB) and tumour infiltrating lymphocytes (TILs). The exploration of non-TMB determinants of active immune response could improve the prognosis prediction and provide guidance for current immunotherapy. METHODS: The transcriptomic and whole exome sequence data in The Cancer Genome Atlas were used to examine the relationship between TMB and exhausted CD8+ T cells (Tex), as an indicator of tumour antigen-specific T cells across nine major cancer types. Computational clustering analysis was performed on 4510 tumours to identify different immune profiles. NanoString gene expression analysis and single cell RNA-seq analysis using fresh human breast cancer were performed for finding validation. FINDINGS: TMB was found to be poorly correlated with active immune response in various cancer types. Patient clustering analysis revealed a group of tumours with abundant Tex but low TMB. In those tumours, we observed significantly higher expression of the stimulator of interferon genes (STING) signalling. Dendritic cells, particularly those of BATF3+ lineage, were also found to be essential for accumulation of Tex within tumours. Mechanistically, loss of genomic and cellular integrity, marked by decreased DNA damage repair, defective replication stress response, and increased apoptosis were shown to drive STING activation. INTERPRETATION: These results highlight that TMB alone does not fully predict tumour immune profiles, with STING signalling compensating for low TMB in non-hypermutated tumours to enhance anti-tumour immunity. Translating these results, STING agonists may benefit patients with non-hypermutated tumours. STING activation may serve as an additional biomarker to predict response to immune checkpoint blockades alongside TMB. Our research also unravelled the interplay between genomic instability and STING activation, informing potential combined chemotherapy targeting the axis of genomic integrity and immunotherapy. FUNDING: City of Hope Christopher Family Endowed Innovation Fund for Alzheimer's Disease and Breast Cancer Research in honor of Vineta Christopher; Breast Cancer Alliance Early Career Investigator Award; National Cancer Institute of the National Institutes of Health under award number R01CA256989 and R01CA240392.

Cephaliverins A-D, four new lignans from Cephalotaxus oliveri Mast.

Four new lignans named cephaliverins A-D (1-4), along with seven known analogues (5-11), were isolated from Cephalotaxus oliveri Mast. Their structures were elucidated on the basis of HR-ESI-MS and NMR analyses, and their absolute configurations were determined by ECD comparison. Cephaliverin A (1), herpetotriol (5) and hedyotol A (6) exhibited moderate antitumor activity against HepG2 and A549 cell lines.

Nanoplasmonic Single-Tumoroid Microarray for Real-Time Secretion Analysis.

Organoid tumor models have emerged as a powerful tool in the fields of biology and medicine as such 3D structures grown from tumor cells recapitulate better tumor characteristics, making these tumoroids unique for personalized cancer research. Assessment of their functional behavior, particularly protein secretion, is of significant importance to provide comprehensive insights. Here, a label-free spectroscopic imaging platform is presented with advanced integrated optofluidic nanoplasmonic biosensor that enables real-time secretion analysis from single tumoroids. A novel two-layer microwell design isolates tumoroids, preventing signal interference, and the microarray configuration allows concurrent analysis of multiple tumoroids. The dual imaging capability combining time-lapse plasmonic spectroscopy and bright-field microscopy facilitates simultaneous observation of secretion dynamics, motility, and morphology. The integrated biosensor is demonstrated with colorectal tumoroids derived from both cell lines and patient samples to investigate their vascular endothelial growth factor A (VEGF-A) secretion, growth, and movement under various conditions, including normoxia, hypoxia, and drug treatment. This platform, by offering a label-free approach with nanophotonics to monitor tumoroids, can pave the way for new applications in fundamental biological studies, drug screening, and the development of therapies.

Double Advantages of Nutrients and Biostimulants Derived from Sewage Sludge by Alkaline Thermal Hydrolysis Process for Agricultural Use: Quality Promotion of Soil and Crop.

Low-carbon alkaline thermal hydrolysis of sewage sludge for the production of high-quality plant-growth-promoting nutrients and biostimulants is a growing concern for sludge resource utilization in agriculture. Thus, this study aims to investigate functional characteristics and soil biochemical effects of sewage sludge-derived nutrients and biostimulants (SS-NB). The content of heavy metals in SS-NB decreased by 47.39-100%, and an increase in soil protease, invertase, and soil nutrient utilization rates are observed in SS-NB groups. SS-NB substituted for chemical fertilizer increased the diversity and evenness of microbial community and reduced the abundance of the soil-borne bacterial genus Arthrobacter. The dominant community of SS-NB100 group is mainly enriched in Microvirga, Ensifer, Novosphingobium, Bosea and Ellin6055, which are principally beneficial symbiotic bacteria of plants and participated in C and N cycles. Moreover, SS-NB reduced the accumulation of Ktedonobacteria and Nitrosospira, which are involved in the production of CO2 and N2O, and also enhanced the coordination of soil microorganisms with enzyme activities and nutrient utilization rate. In conclusion, the results suggest that SS-NB exerts a positive effect on reducing greenhouse gas emissions and preventing soil-borne diseases, and can further enhance collaboration with soil enzyme activity and soil nutrient utilization by stimulating soil microorganisms.

Diterpenolignans and cephalotane diterpenoids from Cephalotaxus oliveri mast. with antitumor activity.

Four undescribed naturally diterpenolignans, and two cephalotane diterpenoids, along with seven known compounds, including two pairs of enantiomers, were isolated from the twigs and leaves of Cephalotaxus oliveri Mast. Their structures were elucidated via spectroscopic data interpretation, chiral-phase HPLC analysis, NMR calculations, and electronic circular dichroism analysis. All the isolated compounds were evaluated for their cytotoxic activities against three kinds of human tumor cell lines. Among them, compound 8 exhibited the most potent activities against MCF-7, HepG2 and A549 cell lines with IC50 values of 2.83, 4.75 and 2.77 µM, respectively.

Ultralong Stability of Ti3 C2 Tx -MXene Dispersion Through Synergistic Regulation of Storage Environment and Defect Capping with Tris-HCl Buffering.

Aqueous MXene dispersion suffers from a bottleneck issue of oxidation, leading to its gradual deterioration and ultimately compromised physicochemical characteristics. Herein, Tris-HCl buffer is employed to stabilize the diluted Ti3 C2 Tx -MXene dispersion (0.05 mg mL-1 ) through the synergy of its potent pH-regulation capability and capping effect toward oxidation-susceptible defects/edges. Tris-HCl functionalized Ti3 C2 Tx maintained its original morphology, structure, and favorable dispersity even after 150 days of aging under naturally aerated conditions. The pH-regulation nature of Tris-HCl is elucidated through solution monitoring of Ti3 C2 Tx dispersion, while the adsorption of Tris-HCl onto defects/edges is revealed by spectral analysis and multi-scale simulations. Tris-HCl at the neutral pH can bind to the negatively charged basal plane of Ti3 C2 Tx via + HTris moiety, while the other moiety (Tris) interacts with the exposed edge-based Ti atoms and/or intrinsic defects, forming a Ti─N bond that prevents MXene from attack by H2 O and O2 . Besides, Tris-HCl stabilized Ti3 C2 Tx exhibited nearly identical capacitive characteristics to its freshly-etched counterpart, indicating the minimal impact of Tris-HCl on electrochemical performance of Ti3 C2 Tx during long-term storage. This study provides practical guidance for stabilizing MXene in their native aqueous dispersion without compromising the inherent properties.

Organ-specific immune setpoints underlie divergent immune profiles across metastatic sites in breast cancer.

Immune composition within the tumor microenvironment (TME) plays a central role in the propensity of cancer to metastasize and to respond to therapy. Previous studies suggested that the metastatic TME is immune suppressed. However, limited accessibility to multiple metastatic sites within patients has made assessment of the immune TME in the context of multi-organ metastases difficult. We utilized a rapid postmortem tissue collection protocol to assess immune composition in numerous sites of breast cancer metastasis and paired tumor-free tissues. Metastases were found to have comparable immune cell densities and composition to paired tumor-free tissues of the same organ type. In contrast, immune cell densities in both metastatic and tumor-free tissues were significantly different between organ types, with lung immune infiltration consistently greater than liver. These immune profiling results were consistent between both flow cytometry and multiplex immunofluorescence-based spatial analysis. Furthermore, we found granulocytes were a predominant tumor-infiltrating immune cell in both lung and liver metastases and these granulocytes made up the majority of PD-L1-expressing cells in many tissue sites. We also identified distinct potential mechanisms of immunosuppression in lung and liver metastases, with lung having increased expression of PD-L1+ antigen-presenting cells and liver having higher numbers of activated regulatory T cells and HLA-DRlow monocytes. Together these results demonstrate that immune contexture of metastases is dictated by organ type, and that immunotherapy strategies may benefit from unique tailoring to tissue-specific features of the immune TME.

PD-L1-expressing tumor-associated macrophages are immunostimulatory and associate with good clinical outcome in human breast cancer.

Tumor-associated macrophages (TAMs) are the predominant cells that express programmed cell death ligand 1 (PD-L1) within human tumors in addition to cancer cells, and PD-L1+ TAMs are generally thought to be immunosuppressive within the tumor immune microenvironment (TIME). Using single-cell transcriptomic and spatial multiplex immunofluorescence analyses, we show that PD-L1+ TAMs are mature and immunostimulatory with spatial preference to T cells. In contrast, PD-L1- TAMs are immunosuppressive and spatially co-localize with cancer cells. Either higher density of PD-L1+ TAMs alone or ratio of PD-L1+/PD-L1- TAMs correlate with favorable clinical outcome in two independent cohorts of patients with breast cancer. Mechanistically, we show that PD-L1 is upregulated during the monocyte-to-macrophage maturation and differentiation process and does not require external IFN-γ stimulus. Functionally, PD-L1+ TAMs are more mature/activated and promote CD8+ T cells proliferation and cytotoxic capacity. Together, our findings reveal insights into the immunological significance of PD-L1 within the TIME.

Emotional labor mediates the associations between self-consciousness and flow in dancers.

Emotional labor has been a focal point in occupational well-being literature, but studies have long overlooked an important group of emotional laborers: performers. This research represents a pioneering effort to examine dancers' adoption of emotional labor strategies, their antecedent of self-consciousness, and the outcome of flow experience. We explored these elements both in the traditional setting of stage dancing and in the novel context of online dance classes without on-site spectators during the COVID-19 pandemic. The results revealed that dancers employed all three common emotional labor strategies: surface acting, deep acting, and expression of naturally felt emotions, with deep acting being the most frequent. In the traditional setting, only the expression of naturally felt emotions mediated the positive effect of private self-consciousness and the negative effect of public self-consciousness on flow experience. In contrast, in the online setting, only private self-consciousness impacted flow through the mediation of deep acting and expression of naturally felt emotions. This exploratory study bridges dramaturgy-originated theories of emotional labor with empirical performing arts research, preliminarily advancing knowledge in the relevant fields of dance education, self-presentation, and flow studies.

Virus Propagation and Cell-Based Colorimetric Quantification.

Zika virus (ZIKV) is a mosquito-borne virus belonging to the genus Flavivirus. ZIKV infection has been associated with congenital brain abnormalities and potentially Guillain-Barré syndrome in adults. Research on ZIKV to understand the disease mechanisms is important to facilitate vaccine and treatment development. The method of quantifying viruses is crucial and fundamental in the field of virology. The focus forming assay (FFA) is a virus quantification assay that detects the viral antigen with antibodies and identifies the infection foci of cells using the peroxidase immunostaining technique. The current study describes the virus propagation and quantification protocol using both 24-well and 96-well (high throughput) formats. Compared with other similar studies, this protocol has further described foci size optimization, which can serve as a guide to expand the use of this assay for other viruses. Firstly, ZIKV propagation is performed in Vero cells for 3 days. The culture supernatant containing ZIKV is harvested and quantitated using the FFA. Briefly, the virus culture is inoculated onto Vero cells and incubated for 2-3 days. Foci formation is then determined after optimized staining processes, including cell fixation, permeabilization, blocking, antibody binding, and incubation with peroxidase substrate. The stained virus foci are visualized using a stereo microscope (manual counting in 24-well format) or software analyzer (automated counting in 96-well format). The FFA provides reproducible, relatively fast results (3-4 days) and is suitable to be used for different viruses, including non-plaque-forming viruses. Subsequently, this protocol is useful for the study of ZIKV infection and could be used to detect other clinically important viruses.

Metabolomics reveals the molecular mechanism of sewage sludge-derived nutrients and biostimulants stimulating resistance enhancement and the redistribution of carbon and nitrogen metabolism in pakchoi cabbage.

Alkaline thermal hydrolysis of sewage sludge to produce high-quality liquid fertilizer that contains protein, amino acid, organic acid and biostimulants is receiving wide attention, however, the impact on plants and environmental risks must be evaluated for sustainable use. In this study, the interactions between sewage sludge-derived nutrients, biostimulants (SS-NB), and pakchoi cabbage were investigated by a combination of phenotypic and metabolic approaches. Compared with SS-NB0 (single chemical fertilizer), SS-NB100, SS-NB50, and SS-NB25 had no effect on crop yield, but the net photosynthetic rate increased from 1.13 % to 9.82 %. In addition, the antioxidant enzyme activity (SOD) increased from 29.60 % to 71.42 %, with decreasing malondialdehyde (MDA) and H2O2 levels by 84.62-92.93 % and 8.62-18.97 %, respectively, indicating positive effects on photosynthetic and antioxidant capacities. Leaf metabolomics revealed that SS-NB100, SS-NB50, and SS-NB25 resulted in up-regulation of amino acid and alkaloid metabolites, down-regulation of carbohydrate metabolites, and up-regulation and down-regulation of organic acid metabolites, which were involved in carbon and nitrogen redistribution. Galactose metabolism was inactivated by SS-NB100, SS-NB50, and SS-NB25, indicating the protective role of SS-NB in oxidative damage in cells. Furthermore, the application of SS-NB also resulted in a significant reduction in heavy metal contents (Cr, Ni, and Pb) and the target hazard quotient (THQ). The THQ values of Cd, Cr, Ni and Pb were <1.0 in SS-NB50, and may be considered as an optimal fertilization strategy. The result provided better understanding of the phenotypic and metabolic changes imposed by SS-NB-replaced chemical fertilizer nitrogen in pakchoi cabbage leaves.

Gene-microRNA Network Analysis Identified Seven Hub Genes in Association with Progression and Prognosis in Non-Small Cell Lung Cancer.

INTRODUCTION: Lung cancer is the leading cause of cancer deaths in the world and is usually divided into non-small cell lung cancer (NSCLC) and small cell lung cancer. NSCLC is dominant and accounts for 85% of the total cases. Currently, the therapeutic method of NSCLC is not so satisfactory, and thus identification of new biomarkers is critical for new clinical therapy for this disease. METHODS: Datasets of miRNA and gene expression were obtained from the NCBI database. The differentially expressed genes (DEGs) and miRNAs (DEMs) were analyzed by GEO2R tools. The DEG-DEM interaction was built via miRNA-targeted genes by miRWalk. Several hub genes were selected via network topological analysis in Cytoscape. RESULTS: A set of 276 genes were found to be significantly differentially expressed in the three datasets. Functional enrichment by the DAVID tool showed that these 276 DEGs were significantly enriched in the term "cancer", with a statistic p-value of 1.9 × 10-5. The subdivision analysis of the specific cancer types indicated that "lung cancer" occupies the largest category with a p-value of 2 × 10-3. Furthermore, 75 miRNAs were shown to be differentially expressed in three representative datasets. A group of 13 DEGs was selected by analysis of the miRNA-gene interaction of these DEGs and DEMs. The investigation of these 13 genes by GEPIA tools showed that eight of them had consistent results with NSCLC samples in the TCGA database. In addition, we applied the KMplot to conduct the survival analysis of these eight genes and found that seven of them have a significant effect on the prognosis survival of patients. We believe that this study could provide effective research clues for the prevention and treatment of non-small cell lung cancer.