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Diversity, a hallmark of G protein-coupled receptor (GPCR) signaling, partly stems from alternative splicing of a single gene generating more than one isoform for a receptor. Additionally, receptor responses to ligands can be attenuated by desensitization upon prolonged or repeated ligand exposure. Both phenomena have been demonstrated and exemplified by the deuterostome tachykinin signaling system, although the role of phosphorylation in desensitization remains a subject of debate. Here, we describe the signaling system for tachykinin-related peptides (TKRPs) in a protostome, mollusk Aplysia. We cloned the Aplysia TKRP precursor, which encodes three TKRPs (apTKRP-1, apTKRP-2a, and apTKRP-2b) containing the FXGXR-amide motif. In situ hybridization and immunohistochemistry showed predominant expression of TKRP mRNA and peptide in the cerebral ganglia. TKRPs and their posttranslational modifications were observed in extracts of central nervous system ganglia using mass spectrometry. We identified two Aplysia TKRP receptors (apTKRPRs), named apTKRPR-A and apTKRPR-B. These receptors are two isoforms generated through alternative splicing of the same gene and differ only in their intracellular C termini. Structure-activity relationship analysis of apTKRP-2b revealed that both C-terminal amidation and conserved residues of the ligand are critical for receptor activation. C-terminal truncates and mutants of apTKRPRs suggested that there is a C-terminal phosphorylation-independent desensitization for both receptors. Moreover, apTKRPR-B also exhibits phosphorylation-dependent desensitization through the phosphorylation of C-terminal Ser/Thr residues. This comprehensive characterization of the Aplysia TKRP signaling system underscores the evolutionary conservation of the TKRP and TK signaling systems, while highlighting the intricacies of receptor regulation through alternative splicing and differential desensitization mechanisms.
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BACKGROUND: Opioids are the most frequently used drugs to treat pain in cancer patients. However opioid analgesics can cause adverse effects and potential drug-drug interaction. RESEARCH DESIGN AND METHODS: This cross-sectional retrospective study analyzed pDDI in 1839 patients with opioid analgesics in a large comprehensive hospital in China from January 1 to 31 December 2022. Three drug interaction databases were used to screen for pDDI including Drugs (U.S.A.), Medscape (U.S.A.), and Drug Assistant of Dingxiangyuan (China). RESULTS: The prevalence of pDDIs among 1839 patients was around 41.27% of 759 patients, and 564 patients (74.31%) with pDDIs were diagnosed with tumor. Further, the total of 275 various pDDIs combinations were identified. The combination of oxycodone with morphine had the most frequent occurrence of 229 times, and its adverse effects mainly related to exacerbate central respiratory depression. While, gender, tumor, number of diagnoses, and the variety of opioid analgesics used were independent risk factors for pDDIs. CONCLUSIONS: Outpatients taking opioid analgesics had a higher incidence of pDDIs. As consequently, optimized monitoring and management of patients taking opioid analgesics is recommended in order to ensure patient medication safety.
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With the increasing number of microRNAs (miRNAs), identifying essential miRNAs has become an important task that needs to be solved urgently. However, there are few computational methods for essential miRNA identification. Here, we proposed a novel framework called Rotation Forest for Essential MicroRNA identification (RFEM) to predict the essentiality of miRNAs in mice. We first constructed 1,264 miRNA features of all miRNA samples by fusing 38 miRNA features obtained from the PESM paper and 1,226 miRNA functional features calculated based on miRNA-target gene interactions. Then, we employed 182 training samples with 1,264 features to train the rotation forest model, which was applied to compute the essentiality scores of the candidate samples. The main innovations of RFEM were as follows: 1) miRNA functional features were introduced to enrich the diversity of miRNA features; 2) the rotation forest model used decision tree as the base classifier and could increase the difference among base classifiers through feature transformation to achieve better ensemble results. Experimental results show that RFEM significantly outperformed two previous models with the AUC (AUPR) of 0.942 (0.944) in three comparison experiments under 5-fold cross validation, which proved the model's reliable performance. Moreover, ablation study was further conducted to demonstrate the effectiveness of the novel miRNA functional features. Additionally, in the case studies of assessing the essentiality of unlabeled miRNAs, experimental literature confirmed that 7 of the top 10 predicted miRNAs have crucial biological functions in mice. Therefore, RFEM would be a reliable tool for identifying essential miRNAs.
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MicroRNAs , Camundongos , Animais , MicroRNAs/genética , Rotação , Biologia Computacional/métodos , Algoritmos , Predisposição Genética para DoençaRESUMO
Aiming at simplifying the network structure of broad learning system (BLS), this article proposes a novel simplification method called compact BLS (CBLS). Groups of nodes play an important role in the modeling process of BLS, and it means that there may be a correlation between nodes. The proposed CBLS not only focuses on the compactness of network structure but also pays closer attention to the correlation between nodes. Learning from the idea of Fused Lasso and Smooth Lasso, it uses the L1 -regularization term and the fusion term to penalize each output weight and the difference between adjacent output weights, respectively. The L1 -regularization term determines the correlation between the nodes and the outputs, whereas the fusion term captures the correlation between nodes. By optimizing the output weights iteratively, the correlation between the nodes and the outputs and the correlation between nodes are attempted to be considered in the simplification process simultaneously. Without reducing the prediction accuracy, finally, the network structure is simplified more reasonably and a sparse and smooth output weights solution is provided, which can reflect the characteristic of group learning of BLS. Furthermore, according to the fusion terms used in Fused Lasso and Smooth Lasso, two different simplification strategies are developed and compared. Multiple experiments based on public datasets are used to demonstrate the feasibility and effectiveness of the proposed methods.
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Neuropeptides with the C-terminal Wamide (Trp-NH2) are one of the last common ancestors of peptide families of eumetazoans and play various physiological roles. In this study, we sought to characterize the ancient Wamide peptides signaling systems in the marine mollusk Aplysia californica, i.e., APGWamide (APGWa) and myoinhibitory peptide (MIP)/Allatostatin B (AST-B) signaling systems. A common feature of protostome APGWa and MIP/AST-B peptides is the presence of a conserved Wamide motif in the C-terminus. Although orthologs of the APGWa and MIP signaling systems have been studied to various extents in annelids or other protostomes, no complete signaling systems have yet been characterized in mollusks. Here, through bioinformatics, molecular and cellular biology, we identified three receptors for APGWa, namely, APGWa-R1, APGWa-R2, and APGWa-R3. The EC50 values for APGWa-R1, APGWa-R2, and APGWa-R3 are 45, 2100, and 2600 nM, respectively. For the MIP signaling system, we predicted 13 forms of peptides, i.e., MIP1-13 that could be generated from the precursor identified in our study, with MIP5 (WKQMAVWa) having the largest number of copies (4 copies). Then, a complete MIP receptor (MIPR) was identified and the MIP1-13 peptides activated the MIPR in a dose-dependent manner, with EC50 values ranging from 40 to 3000 nM. Peptide analogs with alanine substitution experiments demonstrated that the Wamide motif at the C-terminus is necessary for receptor activity in both the APGWa and MIP systems. Moreover, cross-activity between the two signaling systems showed that MIP1, 4, 7, and 8 ligands could activate APGWa-R1 with a low potency (EC50 values: 2800-22,000 nM), which further supported that the APGWa and MIP signaling systems are somewhat related. In summary, our successful characterization of Aplysia APGWa and MIP signaling systems represents the first example in mollusks and provides an important basis for further functional studies in this and other protostome species. Moreover, this study may be useful for elucidating and clarifying the evolutionary relationship between the two Wamide signaling systems (i.e., APGWa and MIP systems) and their other extended neuropeptide signaling systems.
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Aplysia , Neuropeptídeos , Animais , Sequência de Aminoácidos , Moluscos , PeptídeosRESUMO
Neuropeptides are ubiquitous intercellular signaling molecules in the CNS and play diverse roles in modulating physiological functions by acting on specific G-protein coupled receptors (GPCRs). Among them, the elevenin signaling system is now believed to be present primarily in protostomes. Although elevenin was first identified from the L11 neuron of the abdominal ganglion in mollusc Aplysia californica, no receptors have been described in Aplysia, nor in any other molluscs. Here, using two elevenin receptors in annelid Platynereis dumerilii, we found three putative elevenin GPCRs in Aplysia. We cloned the three receptors and tentatively named them apElevR1, apElevR2, and apElevR3. Using an inositol monophosphate (IP1) accumulation assay, we demonstrated that Aplysia elevenin with the disulfide bond activated the three putative receptors with low EC50 values (ranging from 1.2 to 25 nM), supporting that they are true receptors for elevenin. In contrast, elevenin without the disulfide bond could not activate the receptors, indicating that the disulfide bond is required for receptor activity. Using alanine substitution of individual conserved residues other than the two cysteines, we showed that these residues appear to be critical to receptor activity, and the three different receptors had different sensitivities to the single residue substitution. Finally, we examined the roles of those residues outside the disulfide bond ring by removing these residues and found that they also appeared to be important to receptor activity. Thus, our study provides an important basis for further study of the functions of elevenin and its receptors in Aplysia and other molluscs.
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Aplysia , Neuropeptídeos , Animais , Sequência de Aminoácidos , Aplysia/genética , Neuropeptídeos/química , Receptores Acoplados a Proteínas G/química , DissulfetosRESUMO
Influenza infection in children causes a tremendous global burden. In this study, we aimed to investigate the clinical predictors of severe influenza among children. We retrospectively included hospitalized children who had laboratory-confirmed influenza infection and were admitted to a medical center in Taiwan between 2010 and 2018. Severe influenza infection was defined as needing intensive care. We compared demographics, comorbidities, vaccine status and outcomes between patients with severe and nonsevere infection. There were 1030 children hospitalized for influenza infection: 162 patients needed intensive care and 868 patients did not. Multivariable analysis revealed that an age below 2 years (adjusted odds ratio [aOR] 3.31, 95% confidence interval [CI] 2.22-4.95), underlying cardiovascular disease (aOR 1.84, 95% CI 1.04-3.25), neuropsychological (aOR 4.09, 95% CI 2.59-6.45) or respiratory disease (aOR 3.87, 95% CI 1.42-10.60), patchy infiltrates (aOR 2.52, 95% CI 1.29-4.93), pleural effusion (aOR 6.56, 95% CI 1.66-25.91), and invasive bacterial coinfection (aOR 21.89, 95% CI 2.19-218.77) were significant clinical predictors of severe disease, whereas severe infection was less likely in individuals who had received influenza and pneumococcal conjugate vaccines (PCVs) (aOR 0.51, 95% CI 0.28-0.91; aOR 0.35, 95% CI 0.23-0.51, respectively). The most significant risk factors associated with severe influenza infection were an age under 2 years, comorbidities (cardiovascular, neuropsychological, and respiratory diseases), patchy infiltrates or effusion shown on chest X-rays, and bacterial coinfections. The incidence rate of severe disease was significantly lower in those who had received influenza vaccines and PCVs.
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Vacinas contra Influenza , Influenza Humana , Criança , Humanos , Pré-Escolar , Influenza Humana/epidemiologia , Influenza Humana/prevenção & controle , Estudos Retrospectivos , Fatores de Risco , HospitalizaçãoRESUMO
Microalgae-photosynthetic bacteria (PSB) co-culture, which is promising for wastewater treatment and lipid production, is lacking of study. In this work, the combinations of 3 microalgae and 3 PSB strains were firstly screened and then different inoculation ratios of the co-cultures were investigated. It was found the best promotion was Chlorella pyrenoidosa/Rhodobacter capsulatus co-culture (1:1), where the biomass productivity, acetate assimilation rate and lipid productivity were 1.64, 1.61 and 2.79 times than that of the sum of pure microalgae and PSB cultures, respectively. Meanwhile, the inoculation ratio significantly affected the growth rate and lipid productivity of co-culture systems. iTRAQ analysis showed that PSB played a positive effect on acetate assimilation, TCA cycle and glyoxylate cycle of microalgae, but decreased the carbon dioxide utilization and photosynthesis, indicating PSB promoted the microalgae metabolism of organic carbon utilization and weakened inorganic carbon utilization. These findings provide in-depth understanding of carbon utilization in microalgae-PSB co-culture.
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Chlorella , Microalgas , Microalgas/metabolismo , Chlorella/metabolismo , Lipídeos , Biomassa , Bactérias , Águas ResiduáriasRESUMO
Session-based recommendation (SBR) aims to recommend the next items based on anonymous behavior sequences over a short period of time. Compared with other recommendation paradigms, the information available in SBR is very limited. Therefore, capturing the item relations across sessions is crucial for SBR. Recently, many methods have been proposed to learn article transformation relationships over all sessions. Despite their success, these methods may enlarge the impact of noisy interactions and ignore the complex high-order relationship between non-adjacent items. In this study, we propose a self-supervised global context graph neural network (SGC-GNN) to model high-order transition relations between items over all sessions by using virtual context vectors, each of which connects to all items in a given session and enables to collect and propagation information beyond adjacent items. Moreover, to improve the robustness of the proposed model, we devise a contrastive self-supervised learning (SSL) module as an auxiliary task to jointly learn more robust representations of the items in sessions and train the model to fulfill the SBR task. Experimental results on three benchmark datasets demonstrate the superiority of our model over the state-of-the-art (SOTA) methods and validate the effectiveness of context vectors and the self-supervised module.
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BACKGROUND: Timely and complete restoration of blood flow is the most effective intervention for patients with acute myocardial infarction. However, the efficacy is limited by myocardial ischemia-reperfusion (MI/R) injury. PDE4 (phosphodiesterase-4) hydrolyzes intracellular cyclic adenosine monophosphate and it has 4 subtypes A-D. This study aimed to delineate the role of PDE4B (phosphodiesterase-4 subtype B) in MI/R injury. METHODS: Mice were subjected to 30-minute coronary artery ligation, followed by 24-hour reperfusion. Cardiac perfusion was assessed by laser Doppler flow. Vasomotor reactivities were determined in mouse and human coronary (micro-)arteries. RESULTS: Cardiac expression of PDE4B, but not other PDE4 subtypes, was increased in mice following reperfusion. PDE4B was detected primarily in endothelial and myeloid cells of mouse and human hearts. PDE4B deletion strikingly reduced infarct size and improved cardiac function 24-hour or 28-day after MI/R. PDE4B in bone marrow-derived cells promoted MI/R injury and vascular PDE4B further exaggerated this injury. Mechanistically, PDE4B mediated neutrophil-endothelial cell interaction and PKA (protein kinase A)-dependent expression of cell adhesion molecules, neutrophil cardiac infiltration, and release of proinflammatory cytokines. Meanwhile, PDE4B promoted coronary microcirculatory obstruction and vascular permeability in MI/R, without affecting flow restriction-induced thrombosis. PDE4B blockade increased flow-mediated vasodilatation and promoted endothelium-dependent dilatation of coronary arteries in a PKA- and nitric oxide-dependent manner. Furthermore, postischemia administration with piclamilast, a PDE4 pan-inhibitor, improved cardiac microcirculation, suppressed inflammation, and attenuated MI/R injury in mice. Incubation with sera from patients with acute myocardial infarction impaired acetylcholine-induced relaxations in human coronary microarteries, which was abolished by PDE4 inhibition. Similar protection against MI/R-related coronary injury was recapitulated in mice with PDE4B deletion or inhibition, but not with the pure vasodilator, sodium nitroprusside. CONCLUSIONS: PDE4B is critically involved in neutrophil inflammation and microvascular obstruction, leading to MI/R injury. Selective inhibition of PDE4B might protect cardiac function in patients with acute myocardial infarction designated for reperfusion therapy.
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Infarto do Miocárdio , Traumatismo por Reperfusão Miocárdica , Nucleotídeo Cíclico Fosfodiesterase do Tipo 4/genética , Nucleotídeo Cíclico Fosfodiesterase do Tipo 4/metabolismo , Humanos , Inflamação/metabolismo , Microcirculação , Infarto do Miocárdio/metabolismo , Traumatismo por Reperfusão Miocárdica/metabolismo , Neutrófilos/metabolismoRESUMO
The radiative cooling of butterfly wing scales hierarchy has great value in understanding how poikilotherms adapt to the environment and developing bionic materials. However, it remains unclear what the cooling system is like and how the variation of hierarchy affects the cooling efficiency. Therefore, the correlation between the variations of the structure and emissivity of scale hierarchy is thoroughly investigated in Tirumala limniace (Cramer, 1775), whose thermal properties are highly heterogeneous among different wings and regions but similar between males and females. Patterns were deduced from the biological and model simulation experiments. The scale hierarchy varies at the micro- to nanolevel on both surface and section, corresponding to the variating emissivity. Scales on wing veins and margins have large nanostructured units with small lumens and are distinctly thickened, which bring extraordinarily high emissivity. The variations of light and dark scales, respectively, lead to the high emissivity of the middle region of wings and the front wings. Generally, the elevation of the inner surface area and the thickness of the chitin is the key to enhancing the cooling efficiency. For the first time, the effects of the variation of hierarchy toward emissivity of the mid-infrared spectrum are systematically clarified. It is demonstrated that wing scales integrally differentiate in coping with the heterogeneous cooling needs, which may benefit in balancing multifunctions and the development toward the adaptation to the abiotic environment. The study provides insights into the comprehensive thermoregulation system of butterflies and the further development of radiative cooling materials.
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Borboletas , Feminino , Masculino , Animais , Borboletas/fisiologia , Asas de Animais/fisiologia , Regulação da Temperatura Corporal , Adaptação Fisiológica , Temperatura Baixa , PigmentaçãoRESUMO
Non-small cell lung cancer (NSCLC) is considered to be one of the most prevalent and fatal malignancies, with a poor survival rate. Chimeric antigen receptor T cell (CAR-T) cell therapy is one of the most exciting directions in the field of Cellular immunotherapy. Therefore, CAR-T cells that target c-Met have been developed for use in NSCLC therapy and might be a potential therapeutic strategy. The anti c-Met scFv structure was fused with the transmembrane and intracellular domains. Using a lentiviral vector to load the c-Met CAR gene, then transfected the c-Met CAR lentiviral into human T cells to obtain the second generation c-Met CAR-T expressing CARs stably. In vitro co-culture, experiments revealed that CAR-T cells have high proliferative activity and the potential to secrete cytokines (IL-2, TNF-α, and IFN-γ). c-Met CAR-T cells showed special cellular cytotoxicity in LDH release assay. A subcutaneous tumor model in nude mice was used to test the anticancer effectiveness of c-met CAR-T cells in vivo. For c-Met positive NSCLC tissue, according to tumor volume, weight, fluorescence intensity, and immunohistochemical detection, c-Met CAR-T cells had stronger tumor growth suppression compared to untransduced T cells. HE staining revealed that c-Met CAR-T cells did not produced side effects in nude mice. Taken together, we provided useful method to generate c-Met CAR- T cells, which exhibit enhanced cytotoxicity against NSCLC cells in vitro and in vivo. Thus, providing a new therapeutic avenue for treating NSCLC clinically.Highlights (1) c-Met CAR-T capable of stably expressing c-Met CARs were constructed.(2) c-Met CAR-T have strong anti-tumor ability and proliferation ability in vitro.(3) c-Met CAR-T can effectively inhibit the growth of A549 cells subcutaneous xenografts.
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Carcinoma Pulmonar de Células não Pequenas , Neoplasias Pulmonares , Receptores de Antígenos Quiméricos , Animais , Carcinoma Pulmonar de Células não Pequenas/genética , Carcinoma Pulmonar de Células não Pequenas/terapia , Linhagem Celular Tumoral , Humanos , Imunoterapia Adotiva , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/terapia , Camundongos , Camundongos Nus , Receptores de Antígenos Quiméricos/genética , Linfócitos T , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Neuroblastoma is a common childhood cancer with poor prognosis when at its advanced stage. Checkpoint molecule inhibition is successful in treating multiple advanced adult cancers. We investigated PD-L1 and other checkpoint molecule expression to determine their roles in drug resistance and usefulness as targets for drug therapy. We developed three doxorubicin-resistant (DoxR) cell lines from parental cell lines. Matrigel in vitro invasion assays were used to compare invasiveness. Western blot assays were used to compare PD-L1 expression. Immuno-oncology checkpoint protein panels were used to compare concentrations of 17 checkpoint molecules both cellular and soluble. PD-L1 and 12 other checkpoint molecules were present in all cell lysates of each cell line without significantly different levels. Three were solubilized in the media of each cell line. PD-L1 is expressed in all DoxR and parental neuroblastoma cells and may be a potential target for drug therapy although its role in drug resistance remains unclear. Benchmarking checkpoint molecules provides the basis for future studies identifying targets for directed therapy and biomarkers for cancer detection or prognosis.
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BACKGROUND: This study aimed to systematically analyze the association between long-term use of proton pump inhibitors (PPIs) and the risk of gastric cancer (GC). METHODS: We performed a systematic search of articles on the relationship between long-term use of PPIs and the risk of GC from PubMed and EMBASE. We calculated the pooled odds ratio of GC in PPI users compared to non-PPI users using random-effects models. RESULTS: This meta-analysis included 18 studies from 20 different databases with 4348,905 patients enrolled. In the random effects model, we found that an increased risk of GC among PPI users (ORâ =â 1.94; 95% CI [1.43, 2.64]). The long-term use of PPIs compared with histamine-2 receptor antagonist users did not increase the risk of GC (ORâ =â 1.65; 95% CI [0.92, 2.97]). Stratified analysis showed that PPI users had a significantly increased risk of noncardia GC (ORâ =â 2.53; 95% CI [2.03, 3.15]), but had a relatively small relationship with the risk of gastric cardia cancer. (ORâ =â 1.79; 95% CI [1.06, 3.03]). With the extension of PPI use time, the estimated risk value decreases (<1 year: ORâ =â 6.33, 95% CI [3.76, 10.65]; 1-3 years: ORâ =â 1.82, 95% CI [1.30, 2.55]; >3 years: ORâ =â 1.25, 95% CI [1.00, 1.56]). Despite Helicobacter pylori eradication, the long-term use of PPIs did not alter the increased risk of GC (ORâ =â 2.29; 95% CI [1.57, 3.33]). CONCLUSION: Our meta-analysis found that PPI use may be associated with an increased risk of GC. Further research on the causal relationship between these factors is necessary.
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Inibidores da Bomba de Prótons , Neoplasias Gástricas , Humanos , Antagonistas dos Receptores H2 da Histamina/efeitos adversos , Inibidores da Bomba de Prótons/efeitos adversos , Neoplasias Gástricas/induzido quimicamente , Neoplasias Gástricas/epidemiologiaRESUMO
BACKGROUND: Inhibition of the programmed death ligand 1, programmed death 1 pathway has been successfully used for treatment of multiple advanced adult cancers. However, its use in pediatric osteosarcoma is still in its infancy. In this study, we investigated programmed death ligand 1 and other checkpoint molecules' expression to determine the potential usefulness as targets for drug therapy. METHODS: We incubated human wild-type osteosarcoma cells with incremental concentrations of doxorubicin to create a doxorubicin-resistant cell line. Matrigel in vitro invasion assays were used to compare invasiveness. Comparative programmed death ligand 1 expression was evaluated by Western blot assays. An immuno-oncology checkpoint protein panel was used to compare concentrations of 16 other checkpoint molecules. Chi-square tests and Wilcoxon rank-sum tests were used to determine significant differences. RESULTS: A doxorubicin-resistant cell line was successfully created and was significantly more invasive than wild-type cells (0.47 vs 0.07, P < .001). On Western blot assay, doxorubicin-resistant but not wild-type cells expressed programmed death ligand 1. Doxorubicin-resistant cells had significantly higher levels of T-cell immunoglobulin-3 and cluster of differentiation 86 and higher cluster of differentiation 27, cluster of differentiation 40, lymphocyte-activation gene-3, cluster of differentiation 80, programmed death ligand 1, programmed death ligand 2, and inducible T-cell costimulatory expression than wild-type cells. Both lines expressed B- and T-lymphocyte attenuator, cluster of differentiation 28, herpesvirus entry mediator, and programmed death 1. Herpesvirus entry mediator, cluster of differentiation 40, and programmed death ligand 2 were also present in the culture media of both cell lines. CONCLUSION: Doxorubicin-resistant osteosarcoma seems to express higher programmed death ligand 1 than nonresistant wild-type cells. Benchmarking checkpoint molecules may provide the basis for future studies that elucidate pathways of drug resistance and tumor metastasis, biomarkers for cancer prognosis or recurrence, and future targets for directed drug therapy.
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Swarming behavior facilitates pair formation, and therefore mating, in many eusocial termites. However, the physiological adjustments and morphological transformations of the flight muscles involved in flying and flightless insect forms are still unclear. Here, we found that the dispersal flight of the eusocial termite Reticulitermes chinensis Snyder led to a gradual decrease in adenosine triphosphate supply from oxidative phosphorylation, as well as a reduction in the activities of critical mitochondrial respiratory enzymes from preflight to dealation. Correspondingly, using three-dimensional reconstruction and transmission electron microscopy (TEM), the flight muscles were found to be gradually deteriorated during this process. In particular, two tergo-pleural muscles (IItpm5 and III-tpm5) necessary to adjust the rotation of wings for wing shedding behavior were present only in flying alates. These findings suggest that flight muscle systems vary in function and morphology to facilitate the swarming flight procedure, which sheds light on the important role of swarming in successful extension and fecundity of eusocial termites.
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Voo Animal , Isópteros , Animais , Feminino , Isópteros/anatomia & histologia , Isópteros/química , Isópteros/fisiologia , Isópteros/ultraestrutura , Masculino , Microscopia Eletrônica de Transmissão , Músculos/anatomia & histologia , Músculos/química , Músculos/fisiologia , Músculos/ultraestrutura , ReproduçãoRESUMO
To explore the effects of different coated slow-release calcium peroxide on soil microbial characteristics in gleyed paddy field, we set seven treatments by simulation test in a greenhouse, including CK (none calcium peroxide), calcium peroxide powder, calcium peroxide particle and other four coated slow-release calcium peroxide treatments. Samples were collected in same tillering stage of early season rice. The results showed that all the applications of calcium peroxide could improve the concentrations of active soil organic carbon, soil available nutrients, soil microbial biomass, culturable microorganism population, as well as soil enzyme activity. The coated slow-release calcium peroxide had stronger effects than calcium peroxide particles and calcium peroxide powder. The calcium peroxide coated by ethyl cellulose was the most effective, which enhanced soil active organic carbon, soil microbial biomass carbon, nitrogen, phosphorus by 19.4%, 11.4%, 121.5% and 127.2%, soil alkaline hydrolysis nitrogen and available phosphorus by 4.0% and 45.5%, soil culturable bacteria and culturable microorganism population by 137.3% and 113.7%, fungi and actinomyces number were increased by 33.6% and 44.7%. The enzyme activities of invertase, phosphatase, urease as well as catalase were increased by 92.4%, 91.8%, 112.5% and 17.1%, respectively. The results could provide reference for improving gleyed paddy field with coated calcium peroxide.
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Oryza , Solo , Biomassa , Carbono , Nitrogênio , Peróxidos , Microbiologia do SoloRESUMO
Broad learning system (BLS) is a novel neural network with effective and efficient learning ability. BLS has attracted increasing attention from many scholars owing to its excellent performance. This article proposes a weighted BLS (WBLS) based on BLS to tackle the noise and outliers in an industrial process. WBLS provides a unified framework for easily using different methods of calculating the weighted penalty factor. Using the weighted penalty factor to constrain the contribution of each sample to modeling, the normal and abnormal samples were allocated higher and lower weights to increase and decrease their contributions, respectively. Hence, the WBLS can eliminate the bad effect of noise and outliers on the modeling. The weighted ridge regression algorithm is used to compute the algorithm solution. Weighted incremental learning algorithms are also developed using the weighted penalty factor to tackle the noise and outliers in the additional samples and quickly increase nodes or samples without retraining. The proposed weighted incremental learning algorithms provide a unified framework for using different methods of computing weights. We test the feasibility of the proposed algorithms on some public data sets and a real-world application. Experiment results show that our method has better generalization and robustness.
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Objective To prepare and purify human single-chain variable fragment (scFv) targeting c-Met protein and identify its targeting to c-Met protein and its effects on lung adenocarcinoma cells. Methods The scFv gene was inserted into the pFuse vector to construct eukaryotic expression vector that expressed the Met-scFv fused with mouse Fc fragment. Met-scFv was purified by AKTA Protein Purification System and subjected to functional detection. ELISA was used to detect Met-scFv affinity. Flow cytometry and immunofluorescence staining were used to determine the recognition and binding ability of Met-scFv to lung adenocarcinoma cells. The impact of Met-scFv on the proliferation of A549 cells was evaluated by CCK-8 assay. The cytotoxic effect of Met-scFv on A549 cells was examined by the LDH release kit through CDC and ADCC assays. Results ELISA showed that Met-scFv had a dose-effect relationship with c-Met protein. Flow cytometry and immunofluorescence staining revealed that Met-scFv group signal was positive compared with the control group, which suggested that Met-scFv could specifically bind A549 cells. Met-scFv reduced the proliferation of A549 cells and had cytotoxicity to A549 cells, and the toxicity was positively correlated with the dose of Met-scFv. Conclusion Met-scFv targeting c-Met protein has been prepared and it can recognize and mediate to kill A549 cells in vitro.
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Adenocarcinoma de Pulmão , Proteínas Proto-Oncogênicas c-met/antagonistas & inibidores , Anticorpos de Cadeia Única/imunologia , Células A549 , Animais , Ensaio de Imunoadsorção Enzimática , Humanos , CamundongosRESUMO
Objective To establish a hepatocellular carcinoma xenograft model in nude mice which could stably express gene and be monitored dynamically. Methods We first constructed the lentiviral particles containing luciferase (Luc) and near-infrared fluorescent protein (iRFP) and puromycin resistance gene, and then transduced them into the HepG2 hepatoma cells. The cell line stably expressing Luc and iRFP genes were screened and inoculated into nude mice to establish xenograft tumor model. Tumor growth was monitored using in vivo imaging system. HE staining and immunohistochemistry were used to evaluate the pathological features and tumorigenic ability. Results HepG2 cells stably expressing iRFP and Luc were obtained; with the engineered cell line, xenograft model was successfully established with the features of proper tumor developing time and high rate of tumor formation as well as typical pathological features as showed by HE staining and immunohistochemistry. Conclusion Hepatocellular carcinoma model in nude mice with the features of stable gene expression and dynamical monitoring has been established successfully with the HepG2-iRFP-Luc cell line.