The complete chloroplast genome sequences of three lilies: genome structure, comparative genomic and phylogenetic analyses.

We sequenced and analyzed the complete chloroplast genomes of Lilium amoenum, Lilium souliei, and Nomocharis forrestii in detail, including the first sequence and structural comparison of Nomocharis forrestii. We found that the lengths and nucleotide composition of the three chloroplast genes showed little variation. The chloroplast genomes of the three Lilium species contain 87 protein coding genes (PCGs), 38 tRNAs, and 8 rRNA genes. The only difference is that Nomocharis forrestii had an additional infA pseudogene. In the sequence analysis of the Lilium chloroplast genomes, 216 SSRs, 143 pairs of long repeats, 571 SNPs, and 202 indels were detected. In addition, we identified seven hypervariable regions that can be used as potential molecular markers and DNA barcodes of Lilium through complete sequence alignment. The phylogenetic tree was constructed from the three chloroplast genome sequences of Lilium obtained here and 40 chloroplast genome sequences from the NCBI database (including 35 Lilium species, 4 Fritillaria species, and one species of Smilax). The analysis showed that the species clustering of the genus Lilium essentially conformed to the classical morphological classification system of Comber, but differences in the classification of individual species remained. In our report, we support the reclassification of Lilium henryi and Lilium rosthorniiy in the genus Lilium. In general, this study not only provides genome data for three Lilium species, but also provides a comparative analysis of the Lilium chloroplast genomes. These advances will help to identify Lilium species, clarify the phylogenetic analysis of the Lilium genus, and help to solve and improve the disputes and deficiencies in the traditional morphological classification.

Silver nanoparticles induce developmental toxicity via oxidative stress and mitochondrial dysfunction in zebrafish (Danio rerio).

Sliver nanoparticles (AgNPs) are widely used in industry, agriculture, and medicine, potentially resulting in adverse effects on human health and aquatic environments. Here, we investigated the developmental toxicity of zebrafish embryos with acute exposure to AgNPs. Our results demonstrated developmental defects in 4 hpf zebrafish embryos after exposure to different concentrations of AgNPs for 72 h. In addition, RNA-seq profiling of zebrafish embryos after AgNPs treatment. Further Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses showed that the differentially expressed genes (DEGs) were enriched in DNA replication initiation, oxidoreductase activity, DNA replication, cellular senescence, and oxidative phosphorylation signaling pathways in the AgNPs-treated group. Notably, we also found that AgNPs exposure could result in the accumulation of reactive oxygen species (ROS) and malondialdehyde (MDA), the inhibition of superoxide dismutase (SOD), catalase (CAT), and mitochondrial complex I-V activities, and the downregulated expression of SOD, CAT, and mitochondrial complex I-IV chain-related genes. Moreover, the expression of mitochondrion-mediated apoptosis signaling pathway-related genes, such as bax, bcl2, caspase-3, and caspase-9, was significantly regulated after AgNPs exposure in zebrafish. Therefore, these findings demonstrated that AgNPs exposure could cause oxidative stress, induce mitochondrial dysfunction, and ultimately lead to developmental toxicity.

Association between systemic lupus erythematosus and risk of cervical atypia: A meta-analysis.

BACKGROUND: Previous studies showed conflicting results regarding the association between systemic lupus erythematosus (SLE) and risk of cervical atypia. Therefore, the present study aimed to make a meta-analysis to summarize results of studies regarding association between SLE and risk of cervical atypia. METHODS: We searched for articles published before March 2021 in in the following databases: PubMed, EMBASE, Web of Science, Medline and Google Scholar. Odds ratios (ORs) and relative risks (RRs) with their 95% confidence intervals (CIs) were computed to create a pooled effect size and 95% CI using STATA 12.0 software. RESULTS: The present meta-analysis showed that SLE was significantly associated with increased risks of cervical atypia (OR/RR = 2.94, 95% CI 2.22 to 3.89, I2 = 92.1%, p < .001), cervical cancer (OR/RR = 3.13, 95% CI 2.09 to 4.70, I2 = 84.7%, p < .001), squamous intraepithelial lesion (SIL) (OR/RR = 5.00, 95% CI 2.58 to 9.69, I2 = 88.9%, p < .001) and low-grade SIL (OR/RR = 3.14, 95% CI 1.29 to 7.67, I2 = 63.3%, p = .018) with random effects models. CONCLUSION: In summary, findings of this meta-analysis demonstrated that SLE was associated with a higher risk of cervical pre-malignant lesions and carcinoma. It may be necessary for clinicians to remind women with SLE to screen human papillomavirus infection and be vaccinated as soon as possible. However, caution is required when interpreting our findings. Further studies, especially well-designed randomized controlled clinical trials are awaited to confirm the association between SLE and cervical atypia-associated diseases.

Brd4 modulates the innate immune response through Mnk2-eIF4E pathway-dependent translational control of IκBα.

Bromodomain-containing factor Brd4 has emerged as an important transcriptional regulator of NF-κB-dependent inflammatory gene expression. However, the in vivo physiological function of Brd4 in the inflammatory response remains poorly defined. We now demonstrate that mice deficient for Brd4 in myeloid-lineage cells are resistant to LPS-induced sepsis but are more susceptible to bacterial infection. Gene-expression microarray analysis of bone marrow-derived macrophages (BMDMs) reveals that deletion of Brd4 decreases the expression of a significant amount of LPS-induced inflammatory genes while reversing the expression of a small subset of LPS-suppressed genes, including MAP kinase-interacting serine/threonine-protein kinase 2 (Mknk2). Brd4-deficient BMDMs display enhanced Mnk2 expression and the corresponding eukaryotic translation initiation factor 4E (eIF4E) activation after LPS stimulation, leading to an increased translation of IκBα mRNA in polysomes. The enhanced newly synthesized IκBα reduced the binding of NF-κB to the promoters of inflammatory genes, resulting in reduced inflammatory gene expression and cytokine production. By modulating the translation of IκBα via the Mnk2-eIF4E pathway, Brd4 provides an additional layer of control for NF-κB-dependent inflammatory gene expression and inflammatory response.

[Improved Research of Adjustable External Fixation Device for Lower Limbs Based on Semiconductor Refrigeration Sheet].

A kind of adjustable external fixation device for lower extremity is designed. The circuit is mainly composed of TEC1-00703 semiconductor refrigeration chip, HZC-30A pressure sensor, STC89C52RC single chip microcomputer and other electrical components. It can realize the timing intelligent temperature control and meet the local fixed-point refrigeration. The design of adjustable structure and the application of intelligent air cushion can satisfy the full fixation of lower limbs of different individuals. Its operation does not need much medical knowledge. It can solve the problem of emergency transportation and follow-up treatment of lower limb injury in ice and snow sports. It has a good application prospect and universality.

Strong and weak couplings in molecular vibration-plasmon hybrid structures.

Molecular vibration-plasmon couplings in a hybrid structure, which are composed of a silver grating filled with polymethyl methacrylate (PMMA) molecules (SG-PMMA), have been investigated theoretically. It is found that the interaction between the vibrational transitions and plasmons can transform from weak coupling into strong coupling by reducing the distance between the elements. When the space between grating elements is large, the localized surface plasmon resonance (LSP) of the silver elements greatly enhances the absorption of the PMMA molecules. As the gap between elements becomes small, the plasmonic nanocavity (NC) mode emerges and couples strongly with the molecular vibrational mode of PMMA. The strong coupling results in two new hybridized modes and the Rabi splitting energy is about 15 meV. Our work provides an effective way to alter the coupling strength of the molecular vibration-plasmon hybrid system and may be beneficial to the further biochemical and biophysical applications.

Protective effects of ethyl gallate on H2O2-induced mitochondrial dysfunction in PC12 cells.

Oxidative stress has been suggested to play an important role in neuronal injury. Ethyl gallate (EG) is the ethyl ester of gallic acid which has been acknowledged as an antioxidant. We previously demonstrated that EG effectively inhibited H2O2-induced cytotoxicity and decreased the ROS levels in PC12 cells, while the relevant mechanisms of action of this compound remain largely uncharacterized. The present study was carried out in an attempt to clarify the underlying mechanisms of EG against H2O2-induced neurotoxicity in PC12 cells. EG pretreatment attenuated H2O2-induced mitochondrial dysfunction as indicated by the decreased caspase-9/-3 activation, PARP cleavage, mitochondrial membrane potential (MMP) depletion, Bax/Bcl-2 ratio, cytochrome c release and ROS overproduction. Furthermore, EG treatment resulted in nuclear translocation of Nrf2 along with increased expression of ARE-dependent cytoprotective genes, such as γ-GCS and NQO1, which indicated EG as an Nrf2 pathway activator. Silencing of Nrf2 signaling by siRNA abrogated the protective effects offered by EG on H2O2-induced PC12 cells injury, which suggested the important role of Nrf2 pathway in the protection of EG against oxidative stress induced PC12 cell apoptosis. These results taken together indicated that EG protects PC12 cells against H2O2-induced cell mitochondrial dysfunction possibly through activation of Nrf2 pathway. EG might be a potential candidate for further preclinical study aimed at the prevention and treatment of neurodegenerative diseases.

QSAR modelling study of the bioconcentration factor and toxicity of organic compounds to aquatic organisms using machine learning and ensemble methods.

Bioconcentration factors and median lethal concentrations (LC50s) are important when assessing risks posed by organic pollutants to aquatic ecosystems. Various quantitative structure-activity relationship models have been developed to predict bioconcentration factors and classify acute toxicity. In the study, we developed a regression model using Recursive Feature Elimination (RFE) method combined with the Support Vector Machine (SVM) algorithm. We calculated 2D molecular descriptors from a dataset containing 450 diverse chemicals in our regression model. Then we built three ensemble models using three machine learning algorithms and calculated 12 molecular fingerprints from a dataset containing 400 diverse chemicals in our classification models. In the regression model, the R2 and Rpred2 for the regression model were 0.860 and 0.757, respectively. Other parameters indicated that the regression model made good predictions and could efficiently predict a new set of compounds following standards set by Golbraikh, Tropsha, and Roy. In the classification models, the ensemble-SVM classification model gave an overall accuracy, sensitivity, specificity, and area under the receiver operating characteristic curve of 92.2, 95.1, 86.0, and 0.965, respectively, in a five-fold cross-validation and of 87.3, 92.6, 76.0, and 0.940, respectively, in an external validation. These parameters indicated that our ensemble-SVM model was more stable and gave more accurate predictions than previous models. The model could therefore be used to effectively predict aquatic toxicity and assess risks posed to aquatic ecosystems. We identified several structures most relevant to acute aquatic toxicity through predictions made by the two types of models, and this information may be important to aquatic toxicology experiments and aquatic system management.

Bromodomain and extraterminal (BET) protein inhibition suppresses human T cell leukemia virus 1 (HTLV-1) Tax protein-mediated tumorigenesis by inhibiting nuclear factor κB (NF-κB) signaling.

The etiology of human T cell leukemia virus 1 (HTLV-1)-mediated adult T cell leukemia is associated with the ability of viral oncoprotein Tax to induce sustained NF-κB activation and the expression of many NF-κB target genes. Acetylation of the RelA subunit of NF-κB and the subsequent recruitment of bromodomain-containing factor Brd4 are important for the expression of NF-κB target genes in response to various stimuli. However, their contributions to Tax-mediated NF-κB target gene expression and tumorigenesis remain unclear. Here we report that Tax induced the acetylation of lysine 310 of RelA and the binding of Brd4 to acetylated RelA to facilitate Tax-mediated transcriptional activation of NF-κB. Depletion of Brd4 down-regulated Tax-mediated NF-κB target gene expression and cell proliferation. Inhibiting the interaction of Brd4 and acetylated RelA with the bromodomain extraterminal protein inhibitor JQ1 suppressed the proliferation of Tax-expressing rat fibroblasts and Tax-positive HTLV-1-infected cells and Tax-mediated cell transformation and tumorigenesis. Moreover, JQ1 attenuated the Tax-mediated transcriptional activation of NF-κB, triggering the polyubiquitination and proteasome-mediated degradation of constitutively active nuclear RelA. Our results identify Brd4 as a key regulator for Tax-mediated NF-κB gene expression and suggest that targeting epigenetic regulators such as Brd4 with the bromodomain extraterminal protein inhibitor might be a potential therapeutic strategy for cancers and other diseases associated with HTLV-1 infection.

Thermochromic Hydrogels with Adjustable Transition Behavior for Smart Windows.

With the fast economic development and accelerating urbanization, more and more skyscrapers made entirely of concrete and glass are being constructed. To keep a comfortable indoor environment, massive energy for air conditioning or heating appliances is consumed. A huge amount of heat (>30%) is gained or released through glass windows. Using smart windows with the capability to modulate light is an effective way to reduce building energy consumption. Thermochromic hydrogel is one of the potential smart window materials due to its excellent thermal response, high radiation-blocking efficiency, cost-effectiveness, biocompatibility, and good uniformity. In this work, polyhydroxypropyl acrylate (PHPA) hydrogels with controllable lower critical solution temperature (LCST) were prepared by photopolymerization. The transition temperature and transition rate under "static transition" conditions were investigated. Unlike "static" conditions in which the transition temperature was not affected by the initial and final temperature and heating/cooling ramp, the transition temperature varied with the rate of temperature change under dynamic conditions. The "dynamic" transition temperature of the PHPA hydrogel gradually increased with the increase of the heating rate. It was the result of the movement of the molecular chains lagging behind the temperature change when the temperature change was too fast. The results of the solar irradiation experiment by filling PHPA hydrogels into double glazing windows showed that the indoor temperature was about 15 °C lower than that of ordinary glass windows, indicating that it can significantly reduce the energy consumption of air conditioning. In addition, a wide range of adjustable transition temperatures and fast optical response make PHPA hydrogels potentially applicable to smart windows.

Development and Application of a Duplex RT-RPA Assay for the Simultaneous Detection of Cymbidium mosaic virus and Odontoglossum ringspot virus.

Cymbidium mosaic virus (CymMV) and Odontoglossum ringspot virus (ORSV) are among the world's most serious and widespread orchid viruses; they often infect orchids, causing devastating losses to the orchid industry. Therefore, it is critical to establish a method that can rapidly and accurately detect viruses in the field using simple instruments, which will largely reduce the further spread of viruses and improve the quality of the orchid industry and is suitable for mass promotion and application at grassroots agrotechnical service points. In this investigation, we established a rapid amplification method for virus detection at 39 °C for 35 min to detect the presence of CymMV and ORSV simultaneously, sensitively, and specifically in orchids. Primers for the capsid protein (CP)-encoding genes of both viruses were designed and screened, and the reaction conditions were optimized. The experimental amplification process was completed in just 35 min at 39 °C. There were no instances of nonspecific amplification observed when nine other viruses were present. The RPA approach had detection limits of 104 and 103 copies for pMD19T-CymMV and pMD19T-ORSV, respectively. Moreover, the duplex RT-RPA investigation confirmed sensitivity and accuracy via a comparison of detection results from 20 field samples with those of a gene chip. This study presents a precise and reliable detection method for CymMV and ORSV using RT-RPA. The results demonstrate the potential of this method for rapid virus detection. It is evident that this method could have practical applications in virus detection processes.

Development and Validation of a Feature-Based Broad-Learning System for Opportunistic Osteoporosis Screening Using Lumbar Spine Radiographs.

RATIONALE AND OBJECTIVES: Osteoporosis is primarily diagnosed using dual-energy X-ray absorptiometry (DXA); yet, DXA is significantly underutilized, causing osteoporosis, an underdiagnosed condition. We aimed to provide an opportunistic approach to screen for osteoporosis using artificial intelligence based on lumbar spine X-ray radiographs. MATERIALS AND METHODS: In this institutional review board-approved retrospective study, female patients aged ≥50 years who received both X-ray scans and DXA of the lumbar vertebrae, in three centers, were included. A total of 1180 cases were used for training and 145 cases were used for testing. We proposed a novel broad-learning system (BLS) and then compared the performance of BLS models using radiomic features and deep features as a source of input. The deep features were extracted using ResNet18 and VGG11, respectively. The diagnostic performances of these BLS models were evaluated with the area under the curve (AUC), sensitivity, and specificity. RESULTS: The incidence rate of osteoporosis in the training and test sets was 35.9% and 37.9%, respectively. The radiomic feature-based BLS model achieved higher testing AUC (0.802 vs. 0.654 vs. 0.632, both P = .002), sensitivity (78.2% vs. 56.4% vs. 50.9%), and specificity (82.2% vs. 74,4% vs. 75.6%) than the two deep feature-based BLS models. CONCLUSION: Our proposed radiomic feature-based BLS model has the potential to expand osteoporosis screening to a broader population by identifying osteoporosis on lumbar spine X-ray radiographs.

Relating the carbon sources to denitrifying community in full-scale wastewater treatment plants.

Carbon source is a key factor determining the denitrifying effectiveness and efficiency in wastewater treatment plants (WWTPs). Whereas, the relationships between diverse and distinct denitrifying communities and their favorable carbon sources in full-scale WWTPs were not well-understood. This study performed a systematic analysis of the relationships between the denitrifying community and carbon sources by using 15 organic compounds from four categories and activated sludge from 8 full-scale WWTPs. Results showed that, diverse denitrifying bacteria were detected with distinct relative abundances in 8 WWTPs, such as Haliangium (1.98-4.08%), Dechloromonas (2.00-3.01%), Thauera (0.16-1.06%), Zoogloea (0.09-0.43%), and Rhodoferax (0.002-0.104%). Overall, acetate resulted in the highest denitrifying activities (1.21-4.62 mg/L/h/gMLSS), followed by other organic acids (propionate, butyrate and lactate, etc.). Detectable dissimilatory nitrate reduction to ammonium (DNRA) was observed for all 15 carbon sources. Methanol and glycerol resulted in the highest DRNA. Acetate, butyrate, and lactate resulted in the lowest DNRA. Redundancy analysis and 16S cDNA amplicon sequencing suggested that carbon sources within the same category tended to correlate to similar denitrifiers. Methanol and ethanol were primarily correlated to Haliangium. Glycerol and amino acids (glutamate and aspartate) were correlated to Inhella and Sphaerotilus. Acetate, propionate, and butyrate were positively correlated to a wide range of denitrifiers, explaining the high efficiency of these carbon sources. Additionally, even within the same genus, different amplicon sequence variants (ASVs) performed distinctly in terms of carbon source preference and denitrifying capabilities. These findings are expected to benefit carbon source formulation and selection in WWTPs.

Automated and reusable deep learning (AutoRDL) framework for predicting response to neoadjuvant chemotherapy and axillary lymph node metastasis in breast cancer using ultrasound images: a retrospective, multicentre study.

Background: Previous deep learning models have been proposed to predict the pathological complete response (pCR) and axillary lymph node metastasis (ALNM) in breast cancer. Yet, the models often leveraged multiple frameworks, required manual annotation, and discarded low-quality images. We aimed to develop an automated and reusable deep learning (AutoRDL) framework for tumor detection and prediction of pCR and ALNM using ultrasound images with diverse qualities. Methods: The AutoRDL framework includes a You Only Look Once version 5 (YOLOv5) network for tumor detection and a progressive multi-granularity (PMG) network for pCR and ALNM prediction. The training cohort and the internal validation cohort were recruited from Guangdong Provincial People's Hospital (GPPH) between November 2012 and May 2021. The two external validation cohorts were recruited from the First Affiliated Hospital of Kunming Medical University (KMUH), between January 2016 and December 2019, and Shunde Hospital of Southern Medical University (SHSMU) between January 2014 and July 2015. Prior to model training, super-resolution via iterative refinement (SR3) was employed to improve the spatial resolution of low-quality images from the KMUH. We developed three models for predicting pCR and ALNM: a clinical model using multivariable logistic regression analysis, an image model utilizing the PMG network, and a combined model that integrates both clinical and image data using the PMG network. Findings: The YOLOv5 network demonstrated excellent accuracy in tumor detection, achieving average precisions of 0.880-0.921 during validation. In terms of pCR prediction, the combined modelpost-SR3 outperformed the combined modelpre-SR3, image modelpost-SR3, image modelpre-SR3, and clinical model (AUC: 0.833 vs 0.822 vs 0.806 vs 0.790 vs 0.712, all p < 0.05) in the external validation cohort (KMUH). Consistently, the combined modelpost-SR3 exhibited the highest accuracy in ALNM prediction, surpassing the combined modelpre-SR3, image modelpost-SR3, image modelpre-SR3, and clinical model (AUC: 0.825 vs 0.806 vs 0.802 vs 0.787 vs 0.703, all p < 0.05) in the external validation cohort 1 (KMUH). In the external validation cohort 2 (SHSMU), the combined model also showed superiority over the clinical and image models (0.819 vs 0.712 vs 0.806, both p < 0.05). Interpretation: Our proposed AutoRDL framework is feasible in automatically predicting pCR and ALNM in real-world settings, which has the potential to assist clinicians in optimizing individualized treatment options for patients. Funding: National Key Research and Development Program of China (2023YFF1204600); National Natural Science Foundation of China (82227802, 82302306); Clinical Frontier Technology Program of the First Affiliated Hospital of Jinan University, China (JNU1AF-CFTP-2022-a01201); Science and Technology Projects in Guangzhou (202201020022, 2023A03J1036, 2023A03J1038); Science and Technology Youth Talent Nurturing Program of Jinan University (21623209); and Postdoctoral Science Foundation of China (2022M721349).

Adsorption Characterization of Cu(II) and Cd(II) by a Magnetite-Chitosan Composite: Kinetic, Thermodynamic and Equilibrium Studies.

Optimizing the use of magnetite-chitosan composites for heavy metal adsorption has been of great interest due to their environmental friendliness. To gain insights into their potential with green synthesis, this study analyzed one of these composites through X-ray diffraction, Fourier-transform infrared spectroscopy and scanning electron microscopy. Adsorption properties were then explored via static experiments to evaluate the pH dependence, isotherms, kinetics, thermodynamics and regeneration adsorption of Cu(II) and Cd(II). Results disclosed that the optimum pH of adsorption was 5.0, the equilibrium time was about 10 min, and the capacity for Cu(II) and Cd(II) reached 26.28 and 18.67 mg/g, respectively. The adsorption amount of cations increased with temperature from 25 °C to 35 °C and decreased with further increase in temperature from 40 °C to 50 °C, which might be related to the unfolding of chitosan; the adsorption capacity was above 80% of the initial value after two regenerations and about 60% after five regenerations. The composite has a relatively rough outer surface, but its inner surface and porosity are not obvious; it has functional groups of magnetite and chitosan, and chitosan might dominate the adsorption. Consequently, this research proposes the value of maintaining green synthesis research to further optimize the composite system of heavy metal adsorption.

Improving sludge dewaterability by free nitrous acid and lysozyme pretreatment: Performances and mechanisms.

Reducing the water content of waste activated sludge (WAS) is critical for sludge treatment and disposal in wastewater treatment plants (WWTPs). In this study, a new combined conditioning processes by using lysozyme (LZM) and free nitrous acid (FNA) were proposed and demonstrated to enhance the dewaterability of WAS. The water content of sludge cake dropped from 82.82 % to 68.42 % (1 h FNA treatment + 1 h LZM treatment) and 69.52 % (6 h FNA treatment + 1 h LZM treatment) with the combined FNA and LZM treatment; and the corresponding capillary suction time (CST) reduction efficiency increased 49.29 % (1 h FNA treatment + 1 h LZM treatment) and 52.98 % (6 h FNA treatment + 1 h LZM treatment). A comprehensive investigation conducted in this study revealed the underlying mechanism of dewaterability improvement lies in the transformations of extracellular polymeric substances (EPS). The combined conditioning led to enhanced hydrophobicity in the sludge, as suggested by FTIR protein secondary structure and interfacial free energy. The reduced zeta potential and the potential barrier indicated the reduction of the repulsive force of sludge particles and the bound water content in the conditioned floc. The hydrophobicity, flow permeability and flocculability were enhanced after combined treatment, leading to the release of bound water.

Establishment of a triplex TaqMan quantitative real-time PCR assay for simultaneous detection of Cymbidium mosaic virus, Odontoglossum ringspot virus and Cymbidium ringspot virus.

Orchids are significant ornamental plants whose viral infection results in substantial economic damage. Cymbidium mosaic virus (CymMV), Odontoglossum ringspot virus (ORSV), and Cymbidium ringspot virus (CymRSV) represent three important and prevalent orchid viruses. The detection system proposed in this study uses a triplex TaqMan quantitative real-time PCR assay to identify CymMV, ORSV, and CymRSV in a simultaneous manner. We designed specific primers and probes for CymMV, ORSV, and CymRSV, with amplified sequences of 156 bp, 148 bp, and 145 bp, respectively. The minimum detection limit of the triplex qRT-PCR assay for CymMV and CymRSV was 1 copy/assay, and the minimum detection limit was 10 copies/assay for ORSV. The minimum stable detection limits for CymMV, ORSV, and CymRSV were 10, 102, and 102 copies/assay, respectively. Therefore, this system exhibited higher sensitivity (approximately 10 to 104-fold) than RT-PCR. The intra-and interassay CVs of Cq values are less than 0.55 and 0.95%, respectively, indicating that the triplex assay is highly reliable and accurate. In addition, 66 samples from five different orchid genera were analyzed using the established assay and gene chip. The detection results demonstrated that the triplex probe qRT-PCR demonstrated higher sensitivity than the gene chip, indicating that the triplex real-time PCR assay could be used for the detection of field samples. Our findings suggest that the triplex real-time RT-PCR detection system represents a rapid, simple, and accurate tool for detecting CymMV, ORSV, and CymRSV on orchids.

Silver Nanoparticles Cause Neural and Vascular Disruption by Affecting Key Neuroactive Ligand-Receptor Interaction and VEGF Signaling Pathways.

Introduction: Silver nanoparticles (AgNP) are widely used as coating materials. However, the potential risks of AgNP to human health, especially for neural and vascular systems, are still poorly understood. Methods: The vascular and neurotoxicity of various concentrations of AgNP in zebrafish were examined using fluorescence microscopy. In addition, Illumina high-throughput global transcriptome analysis was performed to explore the transcriptome profiles of zebrafish embryos after exposure to AgNP. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were conducted to elucidate the top 3000 differentially expressed genes (DEGs) between AgNP-exposed and control groups. Results: We systematically investigated the neural and vascular developmental toxicities of AgNP exposure in zebrafish. The results demonstrated that AgNP exposure could cause neurodevelopmental anomalies, including a small-eye phenotype, neuronal morphology defects, and inhibition of athletic abilities. In addition, we found that AgNP exposure induces angiogenesis malformation in zebrafish embryos. Further RNA-seq revealed that DEGs were mainly enriched in the neuroactive ligand-receptor interaction and vascular endothelial growth factor (Vegf) signaling pathways in AgNP-treated zebrafish embryos. Specifically, the mRNA levels of the neuroactive ligand-receptor interaction pathway and Vegf signaling pathway-related genes, including si:ch73-55i23.1, nfatc2a, prkcg, si:ch211-132p1.2, lepa, mchr1b, pla2g4aa, rac1b, p2ry6, adrb2, chrnb1, and chrm1b, were significantly regulated in AgNP-treated zebrafish embryos. Conclusion: Our findings indicate that AgNP exposure transcriptionally induces developmental toxicity in neural and vascular development by disturbing neuroactive ligand-receptor interactions and the Vegf signaling pathway in zebrafish embryos.

Nuclear translocation of cGAS orchestrates VEGF-A-mediated angiogenesis.

Cyclic GMP-AMP synthase (cGAS) senses cytosolic incoming DNA and consequently activates stimulator of interferon response cGAMP interactor 1 (STING) to mount immune response. Here, we show nuclear cGAS could regulate VEGF-A-mediated angiogenesis in an immune-independent manner. We found VEGF-A stimulation induces cGAS nuclear translocation via importin-ß pathway. Moreover, nuclear cGAS subsequently regulates miR-212-5p-ARPC3 cascade to modulate VEGF-A-mediated angiogenesis through affecting cytoskeletal dynamics and VEGFR2 trafficking from trans-Golgi network (TGN) to plasma membrane via a regulatory feedback loop. In contrast, cGAS deficiency remarkably impairs VEGF-A-mediated angiogenesis in vivo and in vitro. Furthermore, we found strong association between the expression of nuclear cGAS and VEGF-A, and the malignancy and prognosis in malignant glioma, suggesting that nuclear cGAS might play important roles in human pathology. Collectively, our findings illustrated the function of cGAS in angiogenesis other than immune surveillance, which might be a potential therapeutic target for pathological angiogenesis-related diseases.

Ca-La layered double hydroxide (LDH) for selective and efficient removal of phosphate from wastewater.

Adsorption showed advantages in removing phosphorus (P) at low concentrations. Desirable adsorbents should have sufficiently high adsorption capacity and selectivity. In this study, a Ca-La layered double hydroxide (LDH) was synthesized for the first time by using a simple hydrothermal coprecipitation method for phosphate removal from wastewater. A maximum adsorption capacity of 194.04 mgP/g was achieved, ranking on the top of known LDHs. Adsorption kinetic experiments showed that 0.02 g/L Ca-La LDH could effectively reduce PO43-P from 1.0 to <0.02 mg/L within 30 min. With the copresence of bicarbonate and sulfate at concentrations 17.1 and 35.7 times of that of PO43-P, the Ca-La LDH showed promising selectivity towards phosphate (with a reduction in the adsorption capacity of <13.6%). In addition, four other (Mg-La, Co-La, Ni-La, and Cu-La) LDHs containing different divalent metal ions were synthesized by using the same coprecipitation method. Results showed much higher P adsorption performance of the Ca-La LDH than those LDHs. Field Emission Electron Microscopy (FE-SEM)-Energy Dispersive Spectroscopy (EDS), X-ray Diffraction (XRD), X-ray Photoelectron Spectroscopy (XPS), Fourier Transform Infrared Spectroscopy (FTIR), and mesoporous analysis were performed to characterize and compare the adsorption mechanisms of different LDHs. The high adsorption capacity and selectivity of the Ca-La LDH were mainly explained by selective chemical adsorption, ion exchange, and inner sphere complexation.