Arboviruses antagonize insect Toll antiviral immune signaling to facilitate the coexistence of viruses with their vectors.

Many plant arboviruses are persistently transmitted by piercing-sucking insect vectors. However, it remains largely unknown how conserved insect Toll immune response exerts antiviral activity and how plant viruses antagonize it to facilitate persistent viral transmission. Here, we discover that southern rice black-streaked dwarf virus (SRBSDV), a devastating planthopper-transmitted rice reovirus, activates the upstream Toll receptors expression but suppresses the downstream MyD88-Dorsal-defensin cascade, resulting in the attenuation of insect Toll immune response. Toll pathway-induced the small antibacterial peptide defensin directly interacts with viral major outer capsid protein P10 and thus binds to viral particles, finally blocking effective viral infection in planthopper vector. Furthermore, viral tubular protein P7-1 directly interacts with and promotes RING E3 ubiquitin ligase-mediated ubiquitinated degradation of Toll pathway adaptor protein MyD88 through the 26 proteasome pathway, finally suppressing antiviral defensin production. This virus-mediated attenuation of Toll antiviral immune response to express antiviral defensin ensures persistent virus infection without causing evident fitness costs for the insects. E3 ubiquitin ligase also is directly involved in the assembly of virus-induced tubules constructed by P7-1 to facilitate viral spread in planthopper vector, thereby acting as a pro-viral factor. Together, we uncover a previously unknown mechanism used by plant arboviruses to suppress Toll immune response through the ubiquitinated degradation of the conserved adaptor protein MyD88, thereby facilitating the coexistence of arboviruses with their vectors in nature.

Prediction of lymph node metastasis in operable cervical cancer using clinical parameters and deep learning with MRI data: a multicentre study.

OBJECTIVES: To develop and validate a magnetic resonance imaging-based (MRI) deep multiple instance learning (D-MIL) model and combine it with clinical parameters for preoperative prediction of lymph node metastasis (LNM) in operable cervical cancer. METHODS: A total of 392 patients with cervical cancer were retrospectively enrolled. Clinical parameters were analysed by logistical regression to construct a clinical model (M1). A ResNet50 structure is applied to extract features at the instance level without using manual annotations about the tumour region and then construct a D-MIL model (M2). A hybrid model (M3) was constructed by M1 and M2 scores. The diagnostic performance of each model was evaluated by the area under the receiver operating characteristic curve (AUC) and compared using the Delong method. Disease-free survival (DFS) was evaluated by the Kaplan‒Meier method. RESULTS: SCC-Ag, maximum lymph node short diameter (LNmax), and tumour volume were found to be independent predictors of M1 model. For the diagnosis of LNM, the AUC of the training/internal/external cohort of M1 was 0.736/0.690/0.732, the AUC of the training/internal/external cohort of M2 was 0.757/0.714/0.765, and the AUC of the training/internal/external cohort of M3 was 0.838/0.764/0.835. M3 showed better performance than M1 and M2. Through the survival analysis, patients with higher hybrid model scores had a shorter time to reach DFS. CONCLUSION: The proposed hybrid model could be used as a personalised non-invasive tool, which is helpful for predicting LNM in operable cervical cancer. The score of the hybrid model could also reflect the DFS of operable cervical cancer. CRITICAL RELEVANCE STATEMENT: Lymph node metastasis is an important factor affecting the prognosis of cervical cancer. Preoperative prediction of lymph node status is helpful to make treatment decisions, improve prognosis, and prolong survival time. KEY POINTS: • The MRI-based deep-learning model can predict the LNM in operable cervical cancer. • The hybrid model has the highest diagnostic efficiency for the LNM prediction. • The score of the hybrid model can reflect the DFS of operable cervical cancer.

The impact of threat of shock-induced anxiety on alerting, orienting, and executive function in women: an ERP study.

The present study used a combination of the Threat-of-Shock paradigm and the Attention Network Test (ANT) to investigate how induced anxiety affects alerting, orienting, and executive control and whether individual differences in threat sensitivity moderate these effects. Forty-two female subjects completed the ANT task in alternation under shock-threat and no-shock ("safe") conditions while event-related potentials (ERPs) were recorded. The results showed that anxiety induced by the threat of shock had a significant impact on alerting and executive control functions at the neural level. Specifically, alerting-related N1 and stimulus-preceding negativity (SPN) differences between double cue and no cue conditions were greater in the threat versus safe state, suggesting that the induced anxiety promoted the early perception of cues and preparation for the target. Moreover, executive control-related P3 and sustained potential (SP) differences between incongruent and congruent trials were greater in the threat versus safe state, indicating that the induced anxiety might improve the attentional allocation efficiency and stimulate subjects to recruit more cognitive resources to resolve conflicts. However, orienting-related ERPs were not affected by the threat of shock, but the threat of shock promoted the processing efficiency of spatial-cue at the behavioral level. Analysis of individual differences revealed that trait anxiety moderated the attentional allocation efficiency when performing executive control related tasks in the threat versus safe state. Our findings demonstrate the adaptive significance of the threat of shock-induced anxiety in that being in an anxious state can enhance individuals' alerting, orienting, and executive functions.

Multiregional radiomics of brain metastasis can predict response to EGFR-TKI in metastatic NSCLC.

OBJECTIVES: To develop radiomics signatures from multiparametric magnetic resonance imaging (MRI) scans to detect epidermal growth factor receptor (EGFR) mutations and predict the response to EGFR-tyrosine kinase inhibitors (EGFR-TKIs) in non-small cell lung cancer (NSCLC) patients with brain metastasis (BM). METHODS: We included 230 NSCLC patients with BM treated at our hospital between January 2017 and December 2021 and 80 patients treated at another hospital between July 2014 and October 2021 to form the primary and external validation cohorts, respectively. All patients underwent contrast-enhanced T1-weighted (T1C) and T2-weighted (T2W) MRI, and radiomics features were extracted from both the tumor active area (TAA) and peritumoral edema area (POA) for each patient. The least absolute shrinkage and selection operator (LASSO) was used to identify the most predictive features. Radiomics signatures (RSs) were constructed using logistic regression analysis. RESULTS: For predicting the EGFR mutation status, the created RS-EGFR-TAA and RS-EGFR- POA showed similar performance. By combination of TAA and POA, the multi-region combined RS (RS-EGFR-Com) achieved the highest prediction performance, with AUCs of 0.896, 0.856, and 0.889 in the primary training, internal validation, and external validation cohort, respectively. For predicting response to EGFR-TKI, the multi-region combined RS (RS-TKI-Com) generated the highest AUCs in the primary training (AUC = 0.817), internal validation (AUC = 0.788), and external validation (AUC = 0.808) cohort, respectively. CONCLUSIONS: Our findings suggested values of multiregional radiomics of BM for predicting EGFR mutations and response to EGFR-TKI. CLINICAL RELEVANCE STATEMENT: The application of radiomic analysis of multiparametric brain MRI has proven to be a promising tool to stratify which patients can benefit from EGFR-TKI therapy and to facilitate the precise therapeutics of NSCLC patients with brain metastases. KEY POINTS: • Multiregional radiomics can improve efficacy in predicting therapeutic response to EGFR-TKI therapy in NSCLC patients with brain metastasis. • The tumor active area (TAA) and peritumoral edema area (POA) may hold complementary information related to the therapeutic response to EGFR-TKI. • The developed multi-region combined radiomics signature achieved the best predictive performance and may be considered as a potential tool for predicting response to EGFR-TKI.

Development and validation of MRI-based radiomics signatures as new markers for preoperative assessment of EGFR mutation and subtypes from bone metastases.

BACKGROUND: This study aimed to develop and externally validate contrast-enhanced (CE) T1-weighted MRI-based radiomics for the identification of epidermal growth factor receptor (EGFR) mutation, exon-19 deletion and exon-21 L858R mutation from MR imaging of spinal bone metastasis from primary lung adenocarcinoma. METHODS: A total of 159 patients from our hospital between January 2017 and September 2021 formed a primary set, and 24 patients from another center between January 2017 and October 2021 formed an independent validation set. Radiomics features were extracted from the CET1 MRI using the Pyradiomics method. The least absolute shrinkage and selection operator (LASSO) regression was applied for selecting the most predictive features. Radiomics signatures (RSs) were developed based on the primary training set to predict EGFR mutations and differentiate between exon-19 deletion and exon-21 L858R. The RSs were validated on the internal and external validation sets using the Receiver Operating Characteristic (ROC) curve analysis. RESULTS: Eight, three, and five most predictive features were selected to build RS-EGFR, RS-19, and RS-21 for predicting EGFR mutation, exon-19 deletion and exon-21 L858R, respectively. The RSs generated favorable prediction efficacies for the primary (AUCs, RS-EGFR vs. RS-19 vs. RS-21, 0.851 vs. 0.816 vs. 0.814) and external validation (AUCs, RS-EGFR vs. RS-19 vs. RS-21, 0.807 vs. 0.742 vs. 0.792) sets. CONCLUSIONS: Radiomics features from the CE MRI could be used to detect the EGFR mutation, increasing the certainty of identifying exon-19 deletion and exon-21 L858R mutations based on spinal metastasis MR imaging.

Development and externally validate MRI-based nomogram to assess EGFR and T790M mutations in patients with metastatic lung adenocarcinoma.

OBJECTIVES: This study aims to explore values of multi-parametric MRI-based radiomics for detecting the epidermal growth factor receptor (EGFR) mutation and resistance (T790M) mutation in lung adenocarcinoma (LA) patients with spinal metastasis. METHODS: This study enrolled a group of 160 LA patients from our hospital (between Jan. 2017 and Feb. 2021) to build a primary cohort. An external cohort was developed with 32 patients from another hospital (between Jan. 2017 and Jan. 2021). All patients underwent spinal MRI (including T1-weighted (T1W) and T2-weighted fat-suppressed (T2FS)) scans. Radiomics features were extracted from the metastasis for each patient and selected to develop radiomics signatures (RSs) for detecting the EGFR and T790M mutations. The clinical-radiomics nomogram models were constructed with RSs and important clinical parameters. The receiver operating characteristics (ROC) curve was used to evaluate the predication capabilities of each model. Calibration and decision curve analyses (DCA) were constructed to verify the performance of the models. RESULTS: For detecting the EGFR and T790M mutation, the developed RSs comprised 9 and 4 most important features, respectively. The constructed nomogram models incorporating RSs and smoking status showed favorite prediction efficacy, with AUCs of 0.849 (Sen = 0.685, Spe = 0.885), 0.828 (Sen = 0.964, Spe = 0.692), and 0.778 (Sen = 0.611, Spe = 0.929) in the training, internal validation, and external validation sets for detecting the EGFR mutation, respectively, and with AUCs of 0.0.842 (Sen = 0.750, Spe = 0.867), 0.823 (Sen = 0.667, Spe = 0.938), and 0.800 (Sen = 0.875, Spe = 0.800) in the training, internal validation, and external validation sets for detecting the T790M mutation, respectively. CONCLUSIONS: Radiomics features from the spinal metastasis were predictive on both EGFR and T790M mutations. The constructed nomogram models can be potentially considered as new markers to guild treatment management in LA patients with spinal metastasis. KEY POINTS: • To our knowledge, this study was the first approach to detect the EGFR T790M mutation based on spinal metastasis in patients with lung adenocarcinoma. • We identified 13 MRI features that were strongly associated with the EGFR T790M mutation. • The proposed nomogram models can be considered as potential new markers for detecting EGFR and T790M mutations based on spinal metastasis.

Preparation of CuHY catalyst via solid-state ion exchange method and its catalytic performance in isobutane/2-butene alkylation.

CuHY samples prepared by solid-state ion exchange of HY zeolite with CuCl were used as catalysts in isobutane/2-butene alkylation. The results show that both the addition amount of CuCl and the calcination temperature affect the ion exchange degree. Cu+ can be introduced into Y zeolite by replacing H+ in the HY zeolite after the solid-state ion exchange, causing a decrease of the amount of Brønsted acid sites and an increase of that of Lewis acid sites. When taking CuHY as the catalyst in isobutane/2-butene alkylation, the 3d104s0 valence electron configuration of Cu+ makes it favorable for inhibiting the oligomerization of 2-butene and accelerating the hydride transfer reaction rate by indirectly increasing the local isobutane/olefin ratio around the acid sites of the catalyst. As a result, the selectivity of C8 and trimethylpentanes over CuHY in alkylate are improved compared with those of HY.

Cucurbit Chlorotic Yellows Virus p22 Protein Interacts with Cucumber SKP1LB1 and Its F-Box-Like Motif Is Crucial for Silencing Suppressor Activity.

Plants use RNA silencing as a defense against viruses. In response, viruses encode various RNA silencing suppressors to counteract the antiviral silencing. Here, we identified p22 as a silencing suppressor of cucurbit chlorotic yellows crinivirus and showed that p22 interacts with CsSKP1LB1, a Cucumis sativus ortholog of S-phase kinase-associated protein 1 (SKP1). The F-box-like motif of p22 was identified through sequence analysis and found to be necessary for the interaction using a yeast two-hybrid assay. The involvement of the F-box-like motif in p22 silencing suppressor activity was determined. Proteomics analysis of Nicotiana benthamiana leaves expressing p22, and its F-box-like motif deletion mutant showed 228 differentially expressed proteins and five enriched Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways: ABC transporters, sesquiterpenoid and triterpenoid biosynthesis, ubiquitin-mediated proteolysis, riboflavin metabolism, and cysteine and methionine metabolism. Collectively, our results demonstrate the interaction between p22 and CsSKP1LB1 and show that the deletion of F-box-like motif inhibits p22 silencing suppressor activity. The possible pathways regulated by the p22 through the F-box-like motif were identified using proteomics analysis.

Analgesia-enhancing effects of repetitive transcranial magnetic stimulation on neuropathic pain after spinal cord injury:An fNIRS study.

BACKGROUND: Repetitive transcranial magnetic stimulation (rTMS) is a promising treatment for chronic intractable neuropathic pain in patients with spinal cord injury (SCI). However, the analgesia-enhancing effects of rTMS on conventional interventions (e.g., medications), and the underlying mechanisms remain poorly understood. OBJECTIVE: To investigate the enhancement of analgesia and change of cortex activation by rTMS treatment on neuropathic pain following SCI. METHODS: A double-blind, sham-controlled, clinical trial was performed. Twenty-one patients with neuropathic pain after SCI were randomized (2:1) to receive a session of rTMS (10 Hz, a total of 1200 pulses at an intensity of 80% resting motor threshold) or sham treatment over the left primary motor cortex (M1) corresponding to the hand area daily for six weeks with a one-day interval per week. At T0 (before rTMS treatment), T1 (after the first session rTMS), T2 (after one week), T3 (after two weeks), T4 (after four weeks) and T5 (after six weeks), activations in the bilateral M1, primary somatosensory cortex (S1), premotor cortex (PMC) and prefrontal cortex (PFC) during the handgrip task were measured using functional near-infrared spectroscopy (fNIRS). In addition, the numerical rating scale (NRS) was used to assess pain. RESULTS: The pain intensity or activation in PFC, PMC, M1 or S1 was not remarkably changed at T1. Along with the time, the pain intensity gradually decreased in both the rTMS and sham groups. The real rTMS, compared with the sham, showed more pain relief from two weeks (T3) to six weeks (T5), and the activations of the motor-related areas M1 and PMC were remarkably suppressed. CONCLUSIONS: The findings of this preliminary study with a small patient sample suggest that the analgesia-enhancing effects of high-frequency rTMS might be related with the amelioration of M1 and PMC hypersensitivity, shedding light upon the clinical treatment of SCI-related neuropathic pain.

Notoginsenoside R1 protects WI-38 cells against lipopolysaccharide-triggered injury via adjusting the miR-181a/TLR4 axis.

Notoginsenoside R1 (NGR1) is a neoteric phytoestrogen extracted from Panax notoginseng, and possesses comprehensive pharmacological functions in multitudinous ailments. But, whether NGR1 is utilized in neonatal pneumonia is not clear. This research study aspired to disclose the protective activity of NGR1 in neonatal pneumonia. WI-38 cells were co-stimulated with NGR1 and lipopolysaccharide (LPS, 10 ng/mL), CCK-8 and flow cytometry assays were implemented for cell viability and apoptosis assessment. Real-time quantitative plymerase chain reaction (RT-qPCR), enzyme-linked immunosorbent assay (ELISA), and Western blot analysis were executed for inflammatory cytokine determination. MicroRNA-181a (miR-181a) expression was evaluated through RT-qPCR, simultaneously, the impact of miR-181a was estimated in NGR1 and LPS co-managed cells. Dual luciferase report assay was performed to disclose the relation between miR-181a and Toll-like receptor 4 (TLR4). The nuclear factor-κB (NF-κB) and TAK1/JNK pathways were ultimately appraised. We found that NGR1 decreased cell viability, evoked apoptosis and impeded interleukin-1ß (IL-1ß), IL-6, and tumor necrosis factor-α (TNF-α) expression and secretions in LPS-managed WI-38 cells. MiR-181a expression was enhanced by NGR1, and miR-181a inhibition inverted the impacts of NGR1 in LPS-managed WI-38 cells. Besides, TLR4 was predicted to be a firsthand direct target of miR-181a. Furthermore, NGR1 hindered NF-κB and TAK1/JNK pathways through modulating TLR4. These discoveries disclosed the fact that NGR1 protected WI-38 cells against LPS-triggered injury via adjusting the miR-181a/TLR4 and NF-κB and TAK1/JNK pathways.

Transcriptome analysis of Cucumis sativus infected by Cucurbit chlorotic yellows virus.

BACKGROUND: Cucurbit chlorotic yellows virus (CCYV) is a recently reported bipartite crinivirus that causes chlorotic leaf spots and yellowing symptoms on the leaves of cucurbit plants. The virus-host interaction of CCYV remains to be elucidated, and the influence of criniviruses on the host gene transcriptome requires analysis. METHODS: We used transcriptome sequencing to analyse the differentially expressed genes (DEGs) caused by CCYV infection. RESULTS: CCYV infection resulted in 865 DEGs. The Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis identified 67 pathways, and the three major enrichment pathways (according to the P-values) were photosynthesis-antenna proteins (KO00196), phenylalanine metabolism (KO00360a), and phenylpropanoid biosynthesis (KO00940). Of the 13 DEGs identified in phenylalanine metabolism, 11 genes encode disease resistance-related phenylalanine ammonia-lyase (PAL) genes. Using quantitative real-time PCR, we validated the differential expression of 12 genes. CONCLUSIONS: Our study based on the CCYV-cucumber interaction provides comprehensive transcriptomic information, and will improve our understanding of host-crinivirus interactions.

Infectious clones of the crinivirus cucurbit chlorotic yellows virus are competent for plant systemic infection and vector transmission.

Cucurbit chlorotic yellows virus (CCYV), a recently identified bipartite crinivirus, causes economic losses in cucurbit plants. CCYV is naturally transmitted only by whitefly Bemisia tabaci. Here we constructed full-length cDNA clones of CCYV (RNA1 and RNA2) fused to the T7 RNA polymerase promoter and the cauliflower mosaic virus 35S promoter. CCYV replicated and accumulated efficiently in Cucumis sativus protoplasts transfected with in vitro transcripts. Without RNA2, RNA1 replicated efficiently in C. sativus protoplasts. Agroinoculation with the infectious cDNA clones of CCYV resulted in systemic infection in the host plants of C. sativus and Nicotiana benthamiana. Virus derived from the infectious clones could be transmitted between cucumber plants by vector whiteflies. This system will greatly enhance the reverse genetic studies of CCYV gene functions.

[Effects of injuries pulmonary arterial endothelial cell induced by endotoxin on proliferation of pulmonary artery smooth muscle cells and interference effect of bone morphogenetic protein-2].

OBJECTIVE: To investigate the effects of injuries pulmonary arterial endothelial cell induced by endotoxin on proliferation of pulmonary artery smooth muscle cells and interference effect of bone morphogenetic protein-2, and at the same time explore its possible mechanism. METHODS: Upper liquid of normal PAEC incubated by DMEM for 24 h without serum (EC-CMI) or that of PAEC incubated by the same DMEM and same period after incubating with endotoxin at concentration of 1 microg/ml for 1 h (EM-CMII) were collected and preserved for PASMC incubating. Confluent monolayer PASMC of rats were incubated with EC-CMI (group I), EC-CMII (group II), EC-DMII + BMP-2 1 ng/ml (group III), EC-DMII + BMP-2 10 ng/ml (group IV) and EC-DMII + BMP-2 100 ng/ml (group V) for 24 h. The cultured cells were identified by phase-contrast microscope and immunofluorescent stain of SMC specific antigen. The proliferation of PASMC were evaluated by the BrdU assay. The cell cycle analysis including proliferation index (PI) and S-phase cell fraction (SPF) were performed by the flow cytometry. the DNA synthesis was detected by [methyl-(3)H]-thymidine incorporation; the expression of cyclinD1 mRNA was elucidated by RT-PCR technology; the analysis of phosphorylated Smad1 (pSmad1) and expression of cyclinD1 protein were detected by Western-blot experiment. RESULTS: Cells were confirmed as smooth muscle by their typical "hill-valley" morphological features displayed under phase-contrast microscope and by immunofluorescent staining of smooth muscle, Compared with group I, there were a significant increase in proliferation rate, S-phase cell fraction (SPF), DNA synthesis in group II. Compared with group II, there were no significant changes in group III about that data. But the groups with 10 or 100 ng/ml (group IV and group V) of BMP-2 significantly attenuated proliferation rate, S-phase cell fraction (SPF), DNA synthesis and expression of cyclinD1 mRNA and protein compared with EC-CM2 alone (group II, P < 0.01). Phosphorylated Smad1 increased in group IV and group V compared with group II, (P < 0.01). CONCLUSION: The BMP-2 could downregulate expression of cyclinD1 gene by smad signaling transconduction system, then depressed proliferation of PASMCs by EC-CM2 in vitro culture. And this effect is dependent in the concentration range of 1 - 100 ng/ml.