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The cellular transcription factors are known to play important roles in virus infection. The present study cloned and characterized a transcription factor CCAAT/Enhancer-binding protein homolog from the shrimp Penaeus vannamei (designates as PvCEBP), and explored its potential functions in white spot syndrome virus (WSSV) infection. PvCEBP has an open reading frame (ORF) of 864 bp encoding a putative protein of 287 amino acids, which contained a highly C-terminal conserved bZIP domain. Phylogenetic tree analysis showed that PvCEBP was evolutionarily clustered with invertebrate CEBPs and closely related to the CEBP of Homarus americanus. Quantitative real-time PCR (qPCR) analysis revealed that PvCEBP was expressed in all examined shrimp tissues, with transcript levels increased in shrimp hemocytes and gill upon WSSV challenge. Furthermore, knockdown of PvCEBP mediated by RNA interference significantly decreased the expression of WSSV genes and viral loads, while enhanced the shrimp survival rate under WSSV challenge. In silico prediction and reporter gene assays demonstrated that PvCEBP could activate the promoter activity of the viral immediate-early gene ie1. Collectively, our findings suggest that PvCEBP is annexed by WSSV to promote its propagation by enhancing the expression of viral immediate-early genes.
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Penaeidae , Vírus da Síndrome da Mancha Branca 1 , Animais , Fatores de Transcrição/genética , Penaeidae/genética , Vírus da Síndrome da Mancha Branca 1/fisiologia , Filogenia , Sequência de Aminoácidos , Proteínas de Artrópodes/genéticaRESUMO
For DNA viruses, the immediate-early (IE) proteins are generally essential regulators that manipulate the host machinery to support viral replication. Recently, IE1, an IE protein encoded by white spot syndrome virus (WSSV), has been demonstrated to function as a transcription factor. However, the target genes of IE1 during viral infection remain poorly understood. Here, we explored the host target genes of IE1 using RNAi coupled with transcriptome sequencing analysis. A total of 429 differentially expressed genes (DEGs) were identified from penaeid shrimp, of which 284 genes were upregulated and 145 genes were downregulated after IE1 knockdown. GO and KEGG pathway enrichment analysis revealed the identified DEGs are significantly enriched in the minichromosome maintenance (MCM) complex and DNA replication, indicating that IE1 plays a critical role in DNA replication control. In addition, it was found that Penaeus vannamei MCM complex genes were remarkably upregulated after WSSV infection, while RNAi-mediated knockdown of PvMCM2 reduced the expression of viral genes and viral loads at the early infection stage. Finally, we demonstrated that overexpression of IE1 promoted the expression of MCM complex genes as well as cellular DNA synthesis in insect High-Five cells. Collectively, our current data suggest that the WSSV IE1 protein is a viral effector that modulates the host DNA replication machinery for viral replication.
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Proteínas Imediatamente Precoces , Penaeidae , Vírus da Síndrome da Mancha Branca 1 , Animais , Replicação do DNA/genética , Perfilação da Expressão Gênica , Regulação Viral da Expressão Gênica , Proteínas Imediatamente Precoces/genética , Proteínas Imediatamente Precoces/metabolismo , Penaeidae/metabolismo , Transcriptoma , Vírus da Síndrome da Mancha Branca 1/genéticaRESUMO
The vanilloid-type transient receptor potential (TRPV) channel is reported to be the molecular target of the commercial insecticide pymetrozine, which specifically disrupts the feeding of plant sap-sucking insects. However, the functions of TRPV channels in plant sap-sucking insects have not been fully elucidated. In the present study, RNA interference was used to investigate the effects of the knockdown of TRPV genes (Nan and Iav) on the mortality, locomotion, and feeding behavior of an important plant-feeding insect pest in rice, the brown planthopper, Nilaparvata lugens. Injecting dsRNA of Nan and Iav into fourth-instar nymphs significantly knocked down the target genes. The injection of dsNan or dsIav did not affect any morphological phenotype (including leg extension) of N. lugens nymphs and adults. Knockdown of Nan or Iav resulted in significantly decreased climbing activity against top plants but did not influence the leg-griping strength of adults. Knockdown of Nan resulted in a significantly elevated mortality of N. lugens in the observation period of 7 d after injection, whereas no significant difference in survival rates 7 d after injection was found between dsIav-injected and dsGFP-injected insects. Electropenetrographic (EPG) recordings indicated that knockdown of Nan and Iav reduced the ingestion activity in the rice phloem tissues of N. lugens. Knockdown of Nan and Iav significantly reduced the amount of honeydew excreted by N. lugens. Our findings indicated a relationship between TRPV and N. lugens locomotion and feeding behavior, which may help to fully elucidate the functions of TRPV in insects.
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Hemípteros/genética , Canais de Cátion TRPV/genética , Animais , Comportamento Alimentar , Hemípteros/fisiologia , Locomoção/genética , Ninfa/genética , Ninfa/fisiologia , Oryza , Floema , Interferência de RNA , Canais de Cátion TRPV/metabolismoRESUMO
Tolls and Toll-like receptors (TLRs) are important regulators in the innate immune system and their genetic variations usually affect the host's susceptibility/resistance to pathogen infections. In this study, we report on the single nucleotide polymorphisms (SNPs) of Toll1 in Litopenaeus vannamei (LvToll1) and how this is associated with immune response. PCR-DGGE analysis revealed genetic polymorphisms in LvToll1 at both the genomic DNA (gDNA) and cDNA levels. Using high-throughput sequencing, 223 SNPs were identified at the gDNA level, of which 145 were non-synonymous SNP (nsSNP), with 3 nsSNPs having frequency over 1%. On the other hand, 60 SNPs were identified at the cDNA level including 38 nsSNPs and 4 nsSNPs with frequency over 1%. Upon challenging shrimps with Streptococcus iniae, Vibrio parahaemolyticus and white spot syndrome virus (WSSV), LvToll1 was shown to generate 6, 4 and 4 novel bands, respectively when analyzed with PCR-DGGE. Sequencing analysis of these bands showed that they contained 6, 4 and 2 nsSNPs, respectively. Moreover, the nsSNP C1526T was detected in S. iniae-resistant but not in susceptible shrimps. Most significantly, the C1526T mutation could shorten the α-helix of the LRR domain and was predicted to affect the function of LvToll1, indicating that SNP C1526T might be associated with shrimp's resistance to pathogen infections. In sum, our findings here reveal that the genetic polymorphisms of Toll receptor are linked with the immune response to pathogen infections in L. vannamei.
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Imunidade Inata/genética , Penaeidae/genética , Penaeidae/imunologia , Polimorfismo de Nucleotídeo Único , Receptor 1 Toll-Like/genética , Receptor 1 Toll-Like/imunologia , Sequência de Aminoácidos , Animais , Proteínas de Artrópodes/química , Proteínas de Artrópodes/genética , Proteínas de Artrópodes/imunologia , Perfilação da Expressão Gênica , Alinhamento de Sequência , Receptor 1 Toll-Like/químicaRESUMO
Rhabdomyosarcoma is a highly malignant tumor with striated muscle differentiation, which is histologically classified as alveolar, embryonal, pleomorphic, and spindle cell/sclerosing histological subtype. Rhabdomyosarcoma with TFCP2 rearrangement, which usually occurs in the bone, is a newly identified rare spindle and epithelioid rhabdomyosarcoma with characteristic clinicopathological features and molecular alterations. We report a 39-year-old female patient who underwent local excision of the mandibular lesion. Microscopically, the intraosseous tumor was composed of spindle-shaped, epithelioid, and rhabdomyoblastic cells with atypical nuclei and atypical mitotic figures. In addition, TFCP2 rearrangement was revealed by the fluorescence in situ hybridization. The tumor was thus correctly diagnosed as rhabdomyosarcoma with TFCP2 rearrangement. The patient was scheduled to undergo radiotherapy, and triple-agent chemotherapy after surgery, and no tumor recurrence or metastasis was detected during the 3-month postoperative follow up. Since this tumor is relatively rare and newly recognized, it can be easily misdiagnosed or missed and might be a conundrum of pathological diagnosis. Familiarity with its clinicopathological features and molecular alterations is essential for its correct diagnosis. Therefore, we summarized the clinicopathological, immunohistochemical, and molecular alterations of 43 cases of this rare rhabdomyosarcoma variant in the English-language literature. In addition, the differential diagnosis of this lesion is crucial either.
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RATIONALE AND OBJECTIVES: This study aimed to propose a novel approach of lipiodol combined with drug-eluting beads transarterial chemoembolization (Lipiodol-DEB TACE) and to compare the safety and efficacy with DEB-TACE alone for patients with unresectable hepatocellular carcinoma (HCC). MATERIALS AND METHODS: From the database of four centers, the records of patients with HCC who received DEB-TACE or Lipiodol-DEB TACE as initial treatment were retrospectively evaluated. The tumor response was measured based on the Modified Response Evaluation Criteria in Solid Tumors. Overall survival (OS), progression-free survival (PFS) and adverse events (AEs) were compared between two groups. RESULTS: A total of 244 patients were included with 160 patients receiving DEB-TACE and 84 patients receiving Lipiodol-DEB TACE. Lipiodol-DEB TACE group had higher objective response rate (86.9 % vs. 76.3 %), higher disease control rate (97.6 % vs. 88.8 %), longer median OS (42.6 vs. 25.8 months) and longer median PFS (34.0 vs. 17.0 months) than DEB-TACE group (P < 0.05). There was no significant difference observed in the incidence of AEs between two groups. Cox analysis identified total bilirubin level, maximum tumor diameter, TACE method and portal vein invasion as independent prognostic factors. CONCLUSION: Lipiodol-DEB TACE was a safe option and associated with improved tumor response and survival outcome compared to DEB-TACE alone for selected patients with HCC.
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The excretory organs of insects offer potential physiological targets for insect control. In this study, RNA-seq was utilized to identify a set of transporter and receptor genes enriched in the excretory organs of the brown planthopper (BPH), Nilaparvata lugens, which is considered the most important phloem-feeding insect pest in rice. A total of 1565 and 1084 transcripts were upregulated in the excretory organs, Malpighian tubules, and hindgut, respectively, compared to the midgut, which was enriched for transport activity and oxidoreductase activity. Eight potentially important genes were selected for the exploration of biological function, including one sodium/potassium-ATPase (NKA) subunit (ATP1A1), five aquaporins (AQPs), and two neuropeptide receptors. RNA interference (RNAi) assays showed that the knockdown of ATP1A1 and two AQP genes in BPH resulted in significant lethal phenotypes (corrected mortalities = 42.9-63.6%, 7 days after injection) and significantly reduced honeydew amounts. Our findings suggest that several genes enriched in excretory organs were important for BPH survival, which could be new insect control targets.
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Hemípteros , Oryza , Animais , Transcriptoma , Interferência de RNA , Hemípteros/genética , Oryza/genéticaRESUMO
Objective. Automated medical image segmentation is vital for the prevention and treatment of disease. However, medical data commonly exhibit class imbalance in practical applications, which may lead to unclear boundaries of specific classes and make it difficult to effectively segment certain tail classes in the results of semi-supervised medical image segmentation.Approach. We propose a novel multi-task contrastive learning framework for semi-supervised medical image segmentation with multi-scale uncertainty estimation. Specifically, the framework includes a student-teacher model. We introduce global image-level contrastive learning in the encoder to address the class imbalance and local pixel-level contrastive learning in the decoder to achieve intra-class aggregation and inter-class separation. Furthermore, we propose a multi-scale uncertainty-aware consistency loss to reduce noise caused by pseudo-label bias.Main results. Experiments on three public datasets ACDC, LA and LiTs show that our method achieves higher segmentation performance compared with state-of-the-art semi-supervised segmentation methods.Significance. The multi-task contrastive learning in our method facilitates the negative impact of class imbalance and achieves better classification results. The multi-scale uncertainty estimation encourages consistent predictions for the same input under different perturbations, motivating the teacher model to generate high-quality pseudo-labels. Code is available athttps://github.com/msctransu/MCSSMU.git.
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Processamento de Imagem Assistida por Computador , Humanos , IncertezaRESUMO
Objective.Automatic mutli-organ segmentation from anotomical images is essential in disease diagnosis and treatment planning. The U-shaped neural network with encoder-decoder has achieved great success in various segmentation tasks. However, a pure convolutional neural network (CNN) is not suitable for modeling long-range relations due to limited receptive fields, and a pure transformer is not good at capturing pixel-level features.Approach.We propose a new hybrid network named MSCT-UNET which fuses CNN features with transformer features at multi-scale and introduces multi-task contrastive learning to improve the segmentation performance. Specifically, the multi-scale low-level features extracted from CNN are further encoded through several transformers to build hierarchical global contexts. Then the cross fusion block fuses the low-level and high-level features in different directions. The deep-fused features are flowed back to the CNN and transformer branch for the next scale fusion. We introduce multi-task contrastive learning including a self-supervised global contrast learning and a supervised local contrast learning into MSCT-UNET. We also make the decoder stronger by using a transformer to better restore the segmentation map.Results.Evaluation results on ACDC, Synapase and BraTS datasets demonstrate the improved performance over other methods compared. Ablation study results prove the effectiveness of our major innovations.Significance.The hybrid encoder of MSCT-UNET can capture multi-scale long-range dependencies and fine-grained detail features at the same time. The cross fusion block can fuse these features deeply. The multi-task contrastive learning of MSCT-UNET can strengthen the representation ability of the encoder and jointly optimize the networks. The source code is publicly available at:https://github.com/msctunet/MSCT_UNET.git.
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Redes Neurais de Computação , Software , Processamento de Imagem Assistida por ComputadorRESUMO
White spot syndrome virus (WSSV) is a lethal shrimp pathogen that has a latent infection cycle. The latent virus can easily turn into an acute infection when the culture environment changes, leading to widespread shrimp mortality. However, the mechanism of WSSV latent infection is poorly understood. Bioinformatic analysis revealed that the promoters of WSSV latency-related genes (i.e., wsv151, wsv366, wsv403, and wsv427) contained putative myocyte enhancer factor 2 (MEF2) binding sites. This suggested that the transcription factor MEF2 may be involved in WSSV latent infection. To further investigate this, a MEF2 homolog (PvMEF2) was cloned from Penaeus vannamei and its role in WSSV latent infection was explored. The results showed that knockdown of PvMEF2 led to an increase in the copy number of WSSV, indicating reactivation of WSSV from a latent infection. It was further demonstrated that suppression of PvMEF2 significantly decreased expression of the viral latency-related genes in WSSV-latent shrimp, while overexpression of PvMEF2 in Drosophila S2 cells activated the promoter activity of the viral latency-related gene. Additionally, we demonstrated that silencing of PvMEF2 was able to upregulate the expression of pro-apoptosis genes, thereby promoting cell apoptosis during latent infection. Collectively, the present data suggest that PvMEF2 could promote the expression of virus latency-related genes and enhance cell survival to maintain WSSV latent infection. This finding would contribute to a better understanding of the maintenance mechanism of WSSV latent infection.
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Penaeidae , Vírus da Síndrome da Mancha Branca 1 , Animais , Penaeidae/genética , Penaeidae/metabolismo , Fatores de Transcrição MEF2/genética , Fatores de Transcrição MEF2/metabolismo , Vírus da Síndrome da Mancha Branca 1/genética , Vírus da Síndrome da Mancha Branca 1/metabolismo , Regulação da Expressão Gênica , Regiões Promotoras Genéticas , Drosophila/genéticaRESUMO
Krüppel-like factor 8 (KLF8) regulates critical gene transcription and cellular events associated with cancer. However, KLF8-interacting proteins remain largely unidentified. Using co-immunoprecipitation (co-IP), mass spectrometry, and GST pulldown assays, we identified poly(ADP-ribose) polymerase-1 (PARP-1) as a novel KLF8-interacting protein. Co-IP and Western blotting indicated that KLF8 is also a PARP-1 substrate. Mutation of the cysteines in the zinc finger domain of KLF8 abolished PARP-1 interaction. Surprisingly, immunofluorescent staining revealed a cytoplasmic mislocalization of KLF8 in PARP-1(-/-) cells or when the interaction was disrupted. This mislocalization was prevented by either PARP-1 re-expression or inhibition of CRM1-dependent nuclear export. Interestingly, co-IP indicated competition between PARP-1 and CRM1 for KLF8 binding. Cycloheximide chase assay showed a decrease in the half-life of KLF8 protein when PARP-1 expression was suppressed or KLF8-PARP-1 interaction was disrupted. Ubiquitination assays implicated KLF8 as a target of ubiquitination that was significantly higher in PARP-1(-/-) cells. Promoter reporter assays and chromatin immunoprecipitation assays showed that KLF8 activation on the cyclin D1 promoter was markedly reduced when PARP-1 was deleted or inhibited or when KLF8-PARP-1 interaction was disrupted. Overall, this work has identified PARP-1 as a novel KLF8-binding and -regulating protein and provided new insights into the mechanisms underlying the regulation of KLF8 nuclear localization, stability, and functions.
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Núcleo Celular/metabolismo , Poli(ADP-Ribose) Polimerases/metabolismo , Proteínas Repressoras/metabolismo , Fatores de Transcrição/metabolismo , Transporte Ativo do Núcleo Celular/fisiologia , Animais , Núcleo Celular/genética , Ciclina D1/genética , Ciclina D1/metabolismo , Células HEK293 , Humanos , Carioferinas/genética , Carioferinas/metabolismo , Fatores de Transcrição Kruppel-Like , Camundongos , Camundongos Knockout , Mutação , Poli(ADP-Ribose) Polimerase-1 , Poli(ADP-Ribose) Polimerases/genética , Estabilidade Proteica , Receptores Citoplasmáticos e Nucleares/genética , Receptores Citoplasmáticos e Nucleares/metabolismo , Proteínas Repressoras/genética , Fatores de Transcrição/genética , Dedos de Zinco , Proteína Exportina 1RESUMO
White spot syndrome virus (WSSV) is a large, enveloped, double-stranded DNA virus that threatens shrimp aquaculture worldwide. So far, the mechanisms of WSSV-host interactions are ill-defined. Recent studies have revealed that IE1, an immediate-early protein of WSSV, is a multifunctional modulator implicated in virus-host interactions. In this study, the functions of IE1 were further explored by identifying its interacting proteins using GST-pull down and mass spectrometry analysis. A total of 361 host proteins that potentially bind to IE1 were identified. Bioinformatics analysis revealed that the identified IE1-interactors wereinvolved in various signaling pathways such as prophenoloxidase (proPO) system, PI3K-AKT, and MAPK. Among these, the regulatory role of IE1 in shrimp proPO system was further studied. The Co-immunoprecipitation results confirmed that IE1 interacted with the Ig-like domain of Penaeus vannamei proPO or proPO-like protein (hemocyanin). Additionally, we found that knockdown of IE1 reduced viral genes expression and viral loads and increased the hemocytes' PO activity, whereas recombinant IE1 protein inhibited the PO activity in a dose-dependent manner. Finally, we demonstrated that WSSV could suppress the hemocytes' PO activity at the early infection stage. Collectively, our current data indicate that IE1 is a novel viral regulator that negatively modulates the shrimp proPO system.
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Proteínas Imediatamente Precoces , Vírus da Síndrome da Mancha Branca 1 , Animais , Catecol Oxidase , Precursores Enzimáticos , Proteínas Imediatamente Precoces/genética , Proteínas Imediatamente Precoces/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Proteômica , Vírus da Síndrome da Mancha Branca 1/genética , Vírus da Síndrome da Mancha Branca 1/metabolismoRESUMO
In the Notch signaling pathway in vertebrates and invertebrates, the ligand Delta plays crucial roles in cell proliferation, differentiation, and immunity. Although the Notch signaling pathway has recently been implicated in the immune defense of Penaeus vannamei, the association of Delta with this immune response remains unclear. Here, we cloned and characterized the Delta homolog in P. vannamei (designated as PvDelta). PvDelta has a 2493 bp open reading frame (ORF) encoding a putative protein of 830 amino acids. Bioinformatics analysis revealed that PvDelta contains an N-terminal signal peptide, a conserved Notch ligand (MNNL) domain, a Delta/Serrate/Lag-2 segment, 9 epidermal growth factors segments, a transmembrane domain, and shares high homology with other Delta family members. Transcripts of PvDelta were detected in all shrimp tissues tested and were induced by Vibrio parahaemolyticus, white spot syndrome virus (WSSV), and lipopolysaccharide (LPS), indicating its involvement in shrimp immune response. Moreover, after PvDelta knockdown followed by LPS stimulation, the expression of Notch signaling pathway genes (i.e., PvNotch, PvCSL, and PvHey) was downregulated. Finally, shrimp depleted of PvDelta showed a lower survival rate in response to V. parahaemolyticus challenge. In sum, our data reveal that PvDelta is involved in the innate immunity of shrimp by positively modulating the Notch signaling pathway.
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Proteínas de Artrópodes/metabolismo , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Proteínas de Membrana/metabolismo , Penaeidae/imunologia , Receptores Notch/agonistas , Vibrioses/imunologia , Vibrio parahaemolyticus/fisiologia , Sequência de Aminoácidos , Animais , Proteínas de Artrópodes/genética , Células Cultivadas , Clonagem Molecular , Técnicas de Silenciamento de Genes , Imunidade Inata , Peptídeos e Proteínas de Sinalização Intracelular/genética , Ligantes , Proteínas de Membrana/genética , Alinhamento de Sequência , Transdução de SinaisRESUMO
BACKGROUND: The novel coronavirus disease 2019 (COVID-19) was broken out in December 2019 and soon became a global health emergency. Effective treatment for COVID-19 is urgently needed. In the present study, we aimed to evaluate the antiviral effect of Arbidol vs. Chloroquine in treating COVID-19. METHODS: We retrospectively analyzed 62 patients with COVID-19 diagnosed according to the guidelines for diagnosis and treatment of COVID-19 in China. They were divided into two groups depending on the antiviral drugs that they received. Participants in the Arbidol group (n=42) received 0.2 g Arbidol, tid for 10 days,and those in Chloroquine group (n=20) received 500 mg Chloroquine, bid for 10 days. The coronavirus negative conversion time and the length of hospital stay were analyzed and compared between the two groups. RESULTS: There was no significant difference in demographic and clinical characteristics between the two groups. After antiviral treatment, the nasopharyngeal specimen negative conversion time of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and the length of hospital stay in the Arbidol group were significantly shorter than those in the Chloroquine group (18.50 vs. 25.05 days, P=0.001; 23.52 vs. 28.75 days, P=0.001). Adverse events observed during the antiviral treatment period were comparable between the two groups. Overall, 3 (7.14%) participants in the Arbidol group and 4 (20.0%) in the Chloroquine group experienced adverse events during antiviral treatment. CONCLUSIONS: These results suggest that Arbidol is advantageous over Chloroquine in terms of the SARS-CoV-2 negative conversion and the length of hospital stay in treating COVID-19 patients.
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Antivirais/uso terapêutico , Tratamento Farmacológico da COVID-19 , Cloroquina/uso terapêutico , Indóis/uso terapêutico , China , Cloroquina/efeitos adversos , Humanos , Tempo de Internação , Estudos RetrospectivosRESUMO
Singapore grouper iridovirus (SGIV) is a large double-stranded DNA virus that is a major threat to grouper aquaculture. The pathogenesis of SGIV is not well understood so far. Previous studies have revealed that ICP18, an immediate early protein encoded by SGIV ORF086R gene, promotes viral replication by regulating cell proliferation and virus assembly. In the present study, the potential functions of ICP18 were further explored by probing into its interactors using a proximity-dependent BioID method. Since our in-house grouper embryonic cells (a natural host cell of SGIV) could not be efficiently transfected with the plasmid DNA, and the grouper genome data for mass spectrometry-based protein identification is not currently available, we chosen a non-permissive cell (HEK293 T) as a substitute for this study. A total of 112 cellular proteins that potentially bind to ICP18 were identified by mass spectrometry analysis. Homology analysis showed that among these identified proteins, 110 candidate ICP18-interactors had homologous proteins in zebrafish (a host of SGIV), and shared high sequence identity. Further analysis revealed that the identified ICP18-interacting proteins modulate various cellular processes such as cell cycle and cell adhesion. In addition, the interaction between ICP18 and its candidate interactor, i.e., cyclin-dependent kinase1 (CDK1), was confirmed using Co-immunoprecipitation (Co-IP) and Pull-down assays. Collectively, our present data provides additional insight into the biological functions of ICP18 during viral infection, which could help in further unraveling the pathogenesis of SGIV.
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Bass/virologia , Iridovirus/metabolismo , Proteínas Virais/metabolismo , Animais , Adesão Celular , Ciclo Celular , Quinases Ciclina-Dependentes/metabolismo , Doenças dos Peixes/virologia , Células HEK293 , Humanos , Iridovirus/química , Iridovirus/classificação , Iridovirus/genética , Espectrometria de Massas/métodos , Domínios e Motivos de Interação entre Proteínas , Singapura , Proteínas Virais/genética , Replicação ViralRESUMO
Honeydew is a watery fluid excreted by plant sap-feeding insects. It is a waste product for the insect hosts. However, it plays important roles for other organisms, such as serving as a nutritional source for beneficial insects and bacteria, as well as elicitors and effectors modulating plant responses. In this study, shotgun LC-MS/MS analyses were used to identify the proteins in the honeydew from two important rice hemipteran pests, the brown planthopper (Nilaparvata lugens, BPH) and green rice leafhopper (Nephotettix cincticeps, GRH). A total of 277 and 210 proteins annotated to insect proteins were identified in the BPH and GRH honeydews, respectively. These included saliva proteins that may have similar functions as the saliva proteins, such as calcium-binding proteins and apolipophorin, involved in rice plant defenses. Additionally, a total of 52 and 32 Oryza proteins were identified in the BPH and GRH honeydews, respectively, some of which are involved in the plant immune system, such as Pathogen-Related Protein 10, ascorbate peroxidase, thioredoxin and glutaredoxin. Coincidently, 570 and 494 bacteria proteins were identified from the BPH and GRH honeydews, respectively, which included several well-known proteins involved in the plant immune system: elongation factor Tu, flagellin, GroEL and cold-shock proteins. The results of our study indicate that the insect honeydew is a complex fluid cocktail that contains abundant proteins from insects, plants and microbes, which may be involved in the multitrophic interactions of plants-insects-microbes.
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BACKGROUND: The use of oxaliplatin is very little in the treatment of lung can- cer . This study is designed to find out the efficacy and toxicity of the combination of oxaliplatin and gemcitabine in the treatment of advanced non-small cell lung cancer (NSCLC). METHODS: There were 32 cases of advanced NSCLC patients who were treated with oxaliplatin 65mg/m² by intravenous infusion on the 1st and 8th days, and gemcitabine 800-1000mg/m² by intravenous infusion on the 1st and 8th days every 21 days. After 2 cycles the efficacy was evaluated. RESULTS: There were 31 cases that could be evaluated. No complete response was observed and the overall response rate was 22.6% (7 partial response). The disease control rate was 71.0%. The median time to diease progression was 7 months and the median survival time was 14.5 months. The 1-year survival rate was 59.0% and 2-year survival rate was 45.8%. The main toxicities were myelosuppression and nausea and vomiting. CONCLUSIONS: The combination of oxaliplatin and gemcitabine is effective and well tolerated in the treatment of advanced NSCLC. The quality of life is little influenced and the regimen also can prolong the survival time in some patients.
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In this submission, the phase transition behavior for poly(N-isopropylacrylamide-co-acrylic acid) (pNIPAm-co-AAc) microgels and their assemblies was investigated as a function of temperature and pH using UV-vis spectroscopy (to probe light scattering behavior) and quartz crystal microbalance with dissipation (QCM-D) measurements. PNIPAm-co-AAc microgels were "painted" onto Au-coated glass substrates (for UV-vis) and the Au electrode of a QCM crystal to generate monolayers. The subsequent deposition of another Au layer on top of the pNIPAm-co-AAc microgel layer yields what is known as an etalon. UV-vis/QCM-D measurements revealed that the temperature and pH responsivities for the microgel assemblies match well with their solution behavior. UV-vis spectroscopy shows that the transmittance of the microgel monolayers decreased with increasing solution temperature at pH 3.0. At pH 6.5, the AAc groups in the microgels were deprotonated, leading to strong Coulombic repulsive forces inside the microgels that prevented their collapse and lead to minimal change in the transmitted light intensity. However, QCM-D analysis reveals more complex behavior as it is sensitive to the viscosity/viscoelasticity and thickness changes of the microgel layer, which ultimately depends on the microgel chemical composition and the interaction of the etalon's Au layer with the crystal. The maximum sensitivity to temperature is 0.8 × 10-3 °C·Hz-1, which is the most sensitive pNIPAm microgel-based QCM temperature sensor thus far reported in the literature. Finally, we exploit this new understanding to characterize the pH and ionic strength of a solution using pNIPAm-co-XAAc microgel-based etalon coated crystals. The research results and the sensing demonstration can inspire new and improved sensor designs for a variety of analytes.
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Pemetrexed combined with platinum is a first-line therapy used to treat patients with advanced non-small cell lung cancer (NSCLC) that exhibit negative or unknown epidermal growth factor receptor (EGFR) mutational status or anaplastic lymphoma kinase (ALK) rearrangements. Lung adenocarcinoma (LAC) is the primary type of NSCLC. In order to prevent overtreatment, it is necessary to identify patients with LAC who may not benefit from certain chemotherapies. Patients recruited in the present study (n=129) were diagnosed with advanced LAC and received first-line pemetrexed and platinum-based chemotherapy. A microRNA (miR) microarray was used to screen the plasma miR expression profiles in a screening set of eight patients prior to and following treatment. Specifically, plasma miR-25, miR-21, miR-27b, miR-326, miR-483-5p and miR-920 were selected for reverse transcription-quantitative polymerase chain reaction analysis in a training set (n=44) prior to treatment. The screening and training set patients were all non-smokers with no prior history of serious or chronic disease. The ∆∆Cq values of these miRs were compared between the group that showed benefit from pemetrexed and platinum treatment and the group that did not. Consequently, the ∆∆Cq values of miR-25, miR-21, miR-27b and miR-326 were further determined in a validation set (n=77). The results of the present study demonstrate that plasma expression levels of miR-25, miR-21, miR-27b and miR-326, in the training and validation sets prior to treatment, were significantly different between the benefit and non-benefit groups (P≤0.001). The expression of miR-25, miR-21, miR-27b and miR-326 was upregulated in the non-benefit group and this elevation was positively correlated with decreased progression-free survival (PFS; P≤0.001). In addition, the predictive power of each miR was evaluated through receiver operating characteristic curves, in which miR-25 exhibited the highest degree of accuracy (area under the curve, 0.926; 95% confidence interval, 0.881-0.971). These results indicate that overexpression of plasma miR-25, miR-21, miR-27b and miR-326, prior to treatment, in patients with advanced LAC is predictive of non-benefit from first-line pemetrexed and platinum-based chemotherapy, and is associated with decreased PFS. Among these four miRs, miR-25 exhibited the highest degree of accuracy in predicting insensitivity, suggesting it is the most promising biomarker.
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OBJECTIVE: To study the optimum extraction factors of Buxueyangyan Mixture. METHOD: The orthogonal design was used to select the optimum extraction factors of Cervi Pantotrichum etc. and water extraction rate was used to evaluate the factors. The orthogonal desingn was used to study three factors including crude particle size, extraction time and amount of water. Lcariine content was used as analytical parameter. RESULT: The optimum extraction factors for Cervi Pantotrichum etc. were: 3 a volume ofwith 10 times of water used for three extraction 2.5 h each time. The optimum extraction factors for others were: 1 cm or smaller the crude particle size, boiling 2 times: 2.5, 1.5 h respectively the amount of water used was 10. 8 times of the drug mixture respectively. CONCLUSION: According the optimum extraction factors, the active substance can be extracted efficiently.