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The development of a convergent route to the NLRP3 (nucleotide-binding domain and leucine-rich repeat-containing protein 3) agonist BMS-986299 is reported. The synthesis relies on a key Miyaura borylation and a tandem Suzuki-Miyaura coupling between an iodoimidazole and an o-aminochloroarene, followed by acid-mediated cyclization to afford the aminoquinoline core. The subsequent Boc cleavage and regioselective acylation afford the target compound. Two routes to the iodoimidazole intermediate are presented, along with the synthesis of the o-aminochloroarene via Negishi coupling. The convergent six-step route leads to an 80% reduction in process mass intensity compared to the linear enabling synthesis.
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Imidazóis , Proteína 3 que Contém Domínio de Pirina da Família NLR , Ciclização , AcilaçãoRESUMO
Accurate Force Fields (FFs) are essential for Molecular Dynamics (MD) simulations of the dynamics of realistic materials in terms of atomic-level interactions. The FF parameters of short-range valence interactions can be derived through Quantum Mechanical (QM) calculations on model systems practical for QM (<300 atoms). Similarly, the dynamic electrostatic interactions can be described with methods such as QEq or PQEq that allow charges and polarization to adjust dynamically. However, accurately extracting long-range van der Waals (vdW) interactions from QM calculations poses challenges due to the absence of a definitive method to distinguish between the different energetic components of electrostatics, polarization, vdW, hydrogen bonding, and valence interactions. To do this we use the Perdew-Burke-Ernzerhof flavor of Density Functional Theory, including empirical D3 vdW corrections, to predict the Equation of State for each element (keeping any covalent bonds fixed), from which we obtain the two-body vdW nonbond potential. Here, we extend these calculations to include non-bonded parameters for the N and O columns of the periodic table so that we now describe columns 15 (N), 16 (O), 17 (F), and 18 (Ne) of the periodic table. For these 20 elements, we find that the two-body vdW potentials can all be mapped to a single universal two-body curve, with just three scaling parameters: Re, De, and L. We refer to this as the Universal NonBond (UNB) potential. We expect this to be useful for new MD simulations and a helpful starting point to obtain UNB parameters for the remainder of the periodic table.
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The 1,4-diacyloxylation of 1,3-cyclohexadiene (CHD) affords valuable stereochemically defined scaffolds for natural product and pharmaceutical synthesis. Existing cis-selective diacyloxylation protocols require superstoichiometric quantities of benzoquinone (BQ) or MnO2 , which limit process sustainability and large-scale application. In this report, reaction development and mechanistic studies are described that overcome these limitations by pairing catalytic BQ with tert-butyl hydroperoxide as the stoichiometric oxidant. Catalytic quantities of bromide enable a switch from trans to cis diastereoselectivity. A catalyst with a 1:2 Pd:Br ratio supports high cis selectivity while retaining good rate and product yield. Further studies enable replacement of BQ with hydroquinone (HQ) as a source of cocatalyst, avoiding the handling of volatile and toxic BQ in large-scale applications.
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Acetatos/química , Benzoatos/síntese química , Cicloexenos/síntese química , Hidroquinonas/química , Compostos Organometálicos/química , Catálise , Paládio/química , EstereoisomerismoRESUMO
A Pd-catalyzed ring-opening reaction of 2 H-azirines with carboxylic acids was developed. This reaction undergoes nucleophilic addition between 2,3-diaryl-2 H-azirines and carboxylic acids followed by C-N single-bond cleavage and a subsequent thermal rearrangement. This method enables the rapid construction of valuable α-amido ketone derivatives with high atomic efficiency and superb functional group tolerance.
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OBJECTIVE: The study examines the factors determining the movement time (MT) of positioning an object in an immersive 3D virtual environment. BACKGROUND: Positioning an object into a prescribed area is a fundamental operation in a 3D space. Although Fitts's law models the pointing task very well, it does not apply to a positioning task in an immersive 3D virtual environment since it does not consider the effect of object size in the positioning task. METHOD: Participants were asked to position a ball-shaped object into a spherical area in a virtual space using a handheld or head-tracking controller in the ray-casting technique. We varied object size (OS), movement amplitude (A), and target tolerance (TT). MT was recorded and analyzed in three phases: acceleration, deceleration, and correction. RESULTS: In the acceleration phase, MT was inversely related to object size and positively proportional to movement amplitude. In the deceleration phase, MT was primarily determined by movement amplitude. In the correction phase, MT was affected by all three factors. We observed similar results whether participants used a handheld controller or head-tracking controller. We thus propose a three-phase model with different formulae at each phase. This model fit participants' performance very well. CONCLUSION: A three-phase model can successfully predict MT in the positioning task in an immersive 3D virtual environment in the acceleration, deceleration, and correction phases, separately. APPLICATION: Our model provides a quantitative framework for researchers and designers to design and evaluate 3D interfaces for the positioning task in a virtual space.
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Aceleração , Desaceleração , Atividade Motora/fisiologia , Realidade Virtual , Adulto , Feminino , Humanos , Masculino , Movimento , Fatores de Tempo , Adulto JovemRESUMO
The present study was conducted to investigate the effects of RECK on cervical cancer cell migration and invasion to help understand relevant molecular mechanisms. QRT-PCR and western blot were respectively utilized to examine the transcriptional and translational levels of RECK in cervical cancer cell lines (HELA and C33A) and normal cell line (H8). After transfection with RECK overexpressing vectors, the expression of RECK mRNA, RECK and p53 signaling pathway-related proteins (p21, p53, bcl-2, and Bax) in cervical cancer cells were respectively examined using qRT-PCR and western blot. Cervical cancer cell migration after transfection was detected by wound healing assay and transwell assay. RECK expression was much lower in cervical cancer cell lines compared with normal cell line. Results of wound-healing assay results indicated that RECK could inhibit cervical cancer cell migration, and transwell assay results demonstrated that cell invasion was suppressed by RECK overexpression. Furthermore, western blot indicated that the overexpression of RECK could promote the activation of p53 signaling pathway by influencing related protein expression; whereas its inhibition by PFT-α could antagonize the effect of RECK on migrative and invasive abilities of cervical cancer cells. RECK could inhibit the migration and invasion of cervical cancer cells by activating p53 signaling pathway.
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Regulação para Baixo , Proteínas Ligadas por GPI/genética , Proteínas Ligadas por GPI/metabolismo , Proteína Supressora de Tumor p53/genética , Neoplasias do Colo do Útero/genética , Benzotiazóis/farmacologia , Linhagem Celular Tumoral , Movimento Celular , Feminino , Regulação Neoplásica da Expressão Gênica , Células HeLa , Humanos , Invasividade Neoplásica , Transdução de Sinais , Tolueno/análogos & derivados , Tolueno/farmacologia , Neoplasias do Colo do Útero/metabolismoRESUMO
OBJECTIVE: This study investigated the effects of lncRNA SNHG1 on the proliferation, migration, and invasiveness of cervical cancer cells. METHODS: Three pairs of cervical cancer tissue samples and their corresponding adjacent samples were analyzed using Human LncRNA Microarray V3.0 chip for differential analysis. The expression of SNHG1 in cervical cancer cell lines was verified by qRT-PCR. CCK8 assays and colony formation assays were used to study the changes in cell proliferation. Cell migration and Transwell assays were used to study changes in cell migration and invasiveness. RESULTS: SNHG1 was highly expressed in cervical cancer tissues and cervical cancer cell lines. SNHG1 siRNA could knock-down the expression level of SNHG1 in cervical cancer cell lines HeLa and C33-A. After knock-down of SNHG1, cell proliferation and migration as well as invasiveness in HeLa and C-33A cells decreased. CONCLUSION: LncRNA SNHG1 promotes the development of cervical cancer cells.
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Movimento Celular/efeitos dos fármacos , Invasividade Neoplásica/patologia , RNA Longo não Codificante/farmacologia , Neoplasias do Colo do Útero/patologia , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Feminino , Células HeLa , Humanos , RNA Longo não Codificante/genéticaRESUMO
OBJECTIVE: Fractional exhaled nitric oxide (FeNO) is a non-invasive method used to reveal the eosinophilic inflammation of airway and to diagnose and manage asthma. The use of FeNO measurement or its combination with the small airway function in differentiating CVA patients from chronic cough patients is unexplored. METHODS: From December 2012 to June 2016, patients with chronic non-productive cough for more than 8 weeks based on physician's opinion were subjected to FeNO, pulmonary function, and bronchial hyperresponsiveness (BHR) tests. Receiver operating characteristic (ROC) curves were obtained to evaluate the clinical value of FeNO and small airways indices in CVA diagnosis. The optimal cutoff point for the level of FeNO was also determined. RESULTS: The FeNO value in the CVA group was significantly higher than that in the non-cough-variant asthma (NCVA) group [42(41) parts per billion (ppb) vs. (16(11) ppb), P < 0.05]. The area under the ROC curve was estimated to be 0.874 for FeNO. An optimal cutoff point of 25 ppb had respective sensitivity and specificity values of 81.3% and 84.0% in differentiating CVA patients from NCVA patients. The CVA group showed small airway dysfunction, and the small airway resistance was significantly higher than in the NCVA group. The area under the ROC curve of FeNO combined with maximal midexpiratory flow (MEF) (% predicted) was 0.883, and that combined with MEF50 (% predicted) was 0.886. CONCLUSIONS: The measurement of FeNO is a non-invasive, reproducible, and sensitive method of differentiating CVA patients from NCVA patients. A combination of the level of FeNO (25 ppb) and the abnormal small airway function suggested higher CVA possibility, thereby resulting in a rapid diagnosis. Unnecessary treatments are avoided. This finding provides a new perspective for the management of patients with CVA.
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Asma/diagnóstico , Tosse/diagnóstico , Pulmão/fisiopatologia , Óxido Nítrico/análise , Adulto , Idoso , Asma/fisiopatologia , Testes Respiratórios/métodos , Doença Crônica , Tosse/fisiopatologia , Expiração , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Curva ROC , Reprodutibilidade dos Testes , Adulto JovemRESUMO
We demonstrate the fabrication of an angle-chirped long-period fiber grating (ACLPFG) in a single-mode fiber via CO2 laser pulses. Because of the Berry phase introduced by the ACLPFG, the interference acquires an extra phase difference determined by the torsion of the device. By using that unique characteristic of the proposed device, a high sensitivity sine function torsion response is achieved. The torsion sensitivity is significantly improved, and the temperature crosstalk is effectively avoided by using the relative measurement technology. The torsion sensitivity is ~16 folds (~0.94 nm/ (rad/m)) higher than that of the normal long-period fiber grating (LPFG) with only ~0.006 nm/°C temperature crosstalk within the range of 25-80 °C, which is ~10 folds lower than that of the normal LPFG.
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A novel bidirectional high-sensitivity fiber-optic bending sensor based on the concave-lens-like long-period fiber grating (CLL-LPFG) is designed and demonstrated. The CLL-LPFG is composed by an array of arc-shaped grating planes, and accordingly, its refractive index modulation serves as a concave lens. As a result, the eigencladding mode of the device gets closer to the device surface than the conventional counterpart. Therefore, the proposed sensor provides a more sensitive result. The experimental results show that the bending sensitivities of the CLL-LPFG reach -32.782 nm/m-1 within the bending range of 0-2.08 m-1, which is about sixfold compared to the reported arts. The sensitivity can be potentially improved by optimizing the grating parameters, and the temperature characteristics of the CLL-LPFG can be used to manipulate the grating spectrum.
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A series of structurally new diheteroaryl thioether analogs was designed, prepared and screened toward MGC-803, MKN-45, EC-109 and H1650. Most of the target compounds displayed moderate to potent antiproliferative activities. Among them, compound 5 showed the best antiproliferative activity against the tested cell lines with the half maximal inhibitory concentration (IC50) values below 10µM. In addition, flow cytometry analysis showed that compound 5 increased Bax expression, down-regulated expression of Bcl-2, cleaved caspases-3/9, finally inducing apoptosis of MKN-45 cells as well asarrested the cell cycle at G2/M phase. This study suggests that the diheteroaryl thioethers are a class of emerging chemotypes for developing antitumor agents or biological probes, and compound 5 could serve as a good starting point to design new apoptosis inducers.
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Antineoplásicos/farmacologia , Desenho de Fármacos , Sulfetos/farmacologia , Antineoplásicos/síntese química , Antineoplásicos/química , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Estrutura Molecular , Relação Estrutura-Atividade , Sulfetos/síntese química , Sulfetos/químicaRESUMO
Acetic acid is an inhibitor in industrial processes such as wine making and bioethanol production from cellulosic hydrolysate. It causes energy depletion, inhibition of metabolic enzyme activity, growth arrest and ethanol productivity losses in Saccharomyces cerevisiae. Therefore, understanding the mechanisms of the yeast responses to acetic acid stress is essential for improving acetic acid tolerance and ethanol production. Although 329 genes associated with acetic acid tolerance have been identified in the Saccharomyces genome and included in the database ( http://www.yeastgenome.org/observable/resistance_to_acetic_acid/overview ), the cellular mechanistic responses to acetic acid remain unclear in this organism. Post-genomic approaches such as transcriptomics, proteomics, metabolomics and chemogenomics are being applied to yeast and are providing insight into the mechanisms and interactions of genes, proteins and other components that together determine complex quantitative phenotypic traits such as acetic acid tolerance. This review focuses on these omics approaches in the response to acetic acid in S. cerevisiae. Additionally, several novel strains with improved acetic acid tolerance have been engineered by modifying key genes, and the application of these strains and recently acquired knowledge to industrial processes is also discussed.
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Ácido Acético/farmacologia , Biologia Computacional/métodos , Microbiologia Industrial , Proteínas de Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/efeitos dos fármacos , Tolerância a Medicamentos , Etanol/metabolismo , Fermentação , Perfilação da Expressão Gênica , Regulação Fúngica da Expressão Gênica , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Proteínas de Saccharomyces cerevisiae/genéticaRESUMO
Medical image fusion plays an important role in clinical applications such as image-guided surgery, image-guided radiotherapy, noninvasive diagnosis, and treatment planning. In this paper, we propose a novel multi-modal medical image fusion method based on simplified pulse-coupled neural network and quaternion wavelet transform. The proposed fusion algorithm is capable of combining not only pairs of computed tomography (CT) and magnetic resonance (MR) images, but also pairs of CT and proton-density-weighted MR images, and multi-spectral MR images such as T1 and T2. Experiments on six pairs of multi-modal medical images are conducted to compare the proposed scheme with four existing methods. The performances of various methods are investigated using mutual information metrics and comprehensive fusion performance characterization (total fusion performance, fusion loss, and modified fusion artifacts criteria). The experimental results show that the proposed algorithm not only extracts more important visual information from source images, but also effectively avoids introducing artificial information into fused medical images. It significantly outperforms existing medical image fusion methods in terms of subjective performance and objective evaluation metrics.
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Dissection of the hereditary architecture underlying Saccharomyces cerevisiae tolerance to acetic acid is essential for ethanol fermentation. In this work, a genomics approach was used to dissect hereditary variations in acetic acid tolerance between two phenotypically different strains. A total of 160 segregants derived from these two strains were obtained. Phenotypic analysis indicated that the acetic acid tolerance displayed a normal distribution in these segregants, and suggested that the acetic acid tolerant traits were controlled by multiple quantitative trait loci (QTLs). Thus, 220 SSR markers covering the whole genome were used to detect QTLs of acetic acid tolerant traits. As a result, three QTLs were located on chromosomes 9, 12, and 16, respectively, which explained 38.8-65.9 % of the range of phenotypic variation. Furthermore, twelve genes of the candidates fell into the three QTL regions by integrating the QTL analysis with candidates of acetic acid tolerant genes. These results provided a novel avenue to obtain more robust strains.
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Ácido Acético/metabolismo , Genômica/métodos , Locos de Características Quantitativas , Saccharomyces cerevisiae/crescimento & desenvolvimento , Mapeamento Cromossômico , DNA Fúngico/análise , Repetições de Microssatélites , Fenótipo , Característica Quantitativa Herdável , Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/genéticaRESUMO
Kinetic, spectroscopic, crystallographic, and computational studies probing a Pd-catalyzed C-H arylation reaction reveal that mono-oxidation of the bis-phosphine ligand is critical for the formation of the active catalyst. The bis-phosphine mono-oxide is shown to be a hemilabile, bidentate ligand for palladium. Isolation of the oxidative addition adduct, with structural elucidation by X-ray analysis, showed that the mono-oxide was catalytically competent, giving the same reaction rate in the productive reaction as the Pd(II)/xantphos precursor. A dual role for the carboxylate base in both catalyst activation and reaction turnover was demonstrated, along with the inhibiting effect of excess phosphine ligand. The generality of the role of phosphine mono-oxide complexes in Pd-catalyzed coupling processes is discussed.
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Paládio/química , Fosfinas/química , Xantenos/química , Catálise , Cristalografia por Raios X , Cinética , Espectroscopia de Ressonância Magnética , OxirreduçãoRESUMO
A novel fibrin(ogen)olytic protease from Antheraea pernyi (important economically insect), named cocoonase, was isolated by a combination of ion-exchange chromatography and gel filtration. Furthermore, the characterization of cocoonase was investigated using fibrin(ogen)olytic, thrombolysis, and hemorrhagic assays. The NH2-terminal sequence (IVGGY SVTID KAPYQ) was established by Edman degradation. Based on the N-terminal sequencing, cocoonase cDNA has been cloned by means of RT-PCR and 5'RACE. It is composed of 261 amino acid residues and possesses the structural features of trypsin-like serine protease. The purified cocoonase showed specific esterase activity on N-ß-benzoyl-l-arginine ethyl (BAEE), and the kinetic constants, Km and Vmax were 2.577 × 10(-3)mol/L and 4.09 × 10(-3)µmol/L/s, respectively. Cocoonase showed strong activities on both fibrin and fibrinogen, preferentially hydrolyzed Aα and Bß chains followed by γ-chains of fibrinogen. Cocoonase exhibited a thrombolysis activity both in vitro (blood-clot lysis activity assay) and in vivo (carrageenan-induced thrombosis model). These findings indicate that A. pernyi cocoonase ia a novel fibrin(ogen)olytic enzyme and may have a potential clinical application as an antithrombotic agent.
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Proteínas de Insetos/metabolismo , Mariposas/enzimologia , Peptídeo Hidrolases/metabolismo , Sequência de Aminoácidos , Animais , Arginina/análogos & derivados , Arginina/metabolismo , China , DNA Complementar/química , DNA Complementar/genética , Eletroforese em Gel de Poliacrilamida , Esterases/metabolismo , Fibrina/metabolismo , Fibrinogênio/metabolismo , Fibrinólise , Proteínas de Insetos/administração & dosagem , Proteínas de Insetos/genética , Proteínas de Insetos/isolamento & purificação , Cinética , Camundongos , Dados de Sequência Molecular , Mariposas/genética , Mariposas/metabolismo , Peptídeo Hidrolases/genética , Peptídeo Hidrolases/isolamento & purificação , Filogenia , Análise de Sequência de DNA , Análise de Sequência de Proteína , Homologia de Sequência de Aminoácidos , Especificidade por Substrato , Trombose/metabolismo , Trombose/prevenção & controle , Tripsina/metabolismoRESUMO
BACKGROUND: Neural crest defects lead to congenital heart disease involving outflow tract malformation. Integrin-linked-kinase (ILK) plays important roles in multiple cellular processes and embryogenesis. ILK is expressed in the neural crest, but its role in neural crest and outflow tract morphogenesis remains unknown. RESULTS: We ablated ILK specifically in the neural crest using the Wnt1-Cre transgene. ILK ablation resulted in abnormal migration and overpopulation of neural crest cells in the pharyngeal arches and outflow tract and a significant reduction in the expression of neural cell adhesion molecule (NCAM) and extracellular matrix components. ILK mutant embryos exhibited an enlarged common arterial trunk and ventricular septal defect. Reduced smooth muscle differentiation, but increased ossification and neurogenesis/innervation were observed in ILK mutant outflow tract that may partly be due to reduced transforming growth factor ß2 (TGFß2) but increased bone morphogenetic protein (BMP) signaling. Consistent with these observations, microarray analysis of fluorescence-activated cell sorting (FACS)-sorted neural crest cells revealed reduced expression of genes associated with muscle differentiation, but increased expression of genes of neurogenesis and osteogenesis. CONCLUSIONS: Our results demonstrate that ILK plays essential roles in neural crest and outflow tract development by mediating complex crosstalk between cell matrix and multiple signaling pathways. Changes in these pathways may collectively result in the unique neural crest and outflow tract phenotypes observed in ILK mutants.
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Diferenciação Celular , Movimento Celular , Crista Neural/citologia , Proteínas Serina-Treonina Quinases/metabolismo , Animais , Proteínas Morfogenéticas Ósseas/metabolismo , Adesão Celular , Embrião de Mamíferos , Feminino , Deleção de Genes , Regulação da Expressão Gênica no Desenvolvimento , Camundongos , Camundongos Knockout , Músculo Liso/citologia , Moléculas de Adesão de Célula Nervosa/metabolismo , Crista Neural/metabolismo , Fosforilação , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais , Fator de Crescimento Transformador beta2/metabolismo , Proteína Wnt1/genéticaRESUMO
The Chinese oak silkworm, Antheraea pernyi, is an economically important insect of the Saturniidae family. In this study, genome walking was performed to obtain an A. pernyi actin promoter, which can be employed in transgenic or stable cell line expression systems. The putative promoter was analyzed by the online promoter analysis programs at the Berkeley Drosophila Genome Project and the Web Promoter Scan Service, which led to the recognition of several functional elements. With respect to these elements, a series of actin A1 promoter fragments with 5'-deletions were generated that were then used to construct different vectors expressing Green Fluorescent Protein (GFP). The plasmids were transfected into Sf9 cells and GFP expression was determined by observing GFP fluorescence in cells and by measuring GFP mRNA levels with real-time polymerase chain reaction. Sequence comparisons indicated that the sequence cloned from A. pernyi was the actin A1 promoter. The basic function of the promoter was verified by constructing expression vectors and observing GFP expression. In addition, real-time polymerase chain reaction revealed a strong inhibitory element may exist upstream of the TATA box, which downregulated gene expression. The actin A1 promoter is an ideal candidate for use in A. pernyi transgenic systems.
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Actinas/genética , Proteínas de Insetos/genética , Mariposas/genética , Regiões Promotoras Genéticas , Sequência de Aminoácidos , Animais , Sequência de Bases , Dados de Sequência Molecular , Análise de Sequência de DNARESUMO
OBJECTIVE: To explore the anti-tumor effect and its mechanism of Sendai virus Tianjin strain defective interfering particles (DIP) on mouse models of colon carcinoma. METHODS: CT26 cells (5×10(6)/0.1 ml) were subcutaneously injected into the back of Bal B/c mice to establish murine colon carcinoma model. After the tumors reached 5 mm in diameter, the mice were randomly divided into Tianjin strain DIP group and saline control group. The former was intratumorally injected with Tianjin strain DIP (0.1 ml) once a day on day 4, 7, 10 and 13 after CT26 cell inoculation. The latter was intratumorally injected with the same volume of saline. Tumor volume and survival rate of the mice were calculated to confirm the anti-tumor effect of DIP. Flow cytometry and ELISA were used to examine the maturation and release of cytokines IL-6, IFN-α and TNF-α from murine myeloid dendritic cells (DCs) induced by Tianjin strain DIP. Moreover, real-time RT-PCR and immunohistochemistry were performed to identify whether the Tianjin strain DIP could induce infiltration of CD11c(+) DCs, CD4(+) and CD8(+) T cells in the tumors. RESULTS: On day 22 after CT26 cell inoculation, the average tumor volume of the Tianjin strain DIP group was (33.2 ± 2.0) mm(3), significantly smaller than that of the control group [(2 376.0 ± 130.8)mm(3), P < 0.01]. On day 50 after CT26 cell inoculation, the survival rate of mice was 90.0% in the Tianjin strain DIP group, much higher than that of the control group (30.0%, P < 0.01). Flow cytometry analysis showed that the expression of markers of DCs maturation, including CD40, CD80 and CD86, was dose-dependently increased by DIP or intact virus. No statistically significant difference was found betweent the DIP and intact virus groups. ELISA results showed that DIP could stimulate the secretion of IL-6, IFN-α and TNF-α from mouse DCs. The secretion of all of the cytokines was dose-dependently increased by DIP or intact virus. Real-time RT-PCR revealed that the expression of CD4, CD8 and CD11c mRNAs was increased in tumors treated with DIP compared with that of the saline group at all time points. Moreover, the expression level of all of them remained maximal at 120 h after the last treatment. Immunohistochemical staining revealed that the ratios of CD4(+), CD8(+) T cells or CD11c(+) DCs to total cells were (21.60 ± 1.49)%, (22.12 ± 2.84)% and (23.05 ± 2.91)%, respectively, in the DIP-treated tumors. In the tumors treated by saline, the ratios were (2.62 ± 0.60)%, (4.05 ± 0.12)% and (3.10 ± 0.09)%, respectively. The difference between experimental group and control group had statistical significance. CONCLUSIONS: Tianjin strain DIP may exert anti-tumor effect on tumor-bearing mice. The mechanism is related with the antitumor immunity induced by DCs and T cells.
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Neoplasias do Colo/patologia , Vírus Defeituosos/imunologia , Células Dendríticas/metabolismo , Vírus Sendai/imunologia , Animais , Linhagem Celular Tumoral , Neoplasias do Colo/metabolismo , Citocinas/metabolismo , Feminino , Interferon-alfa/metabolismo , Interleucina-6/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Transplante de Neoplasias , Distribuição Aleatória , Linfócitos T/metabolismo , Carga Tumoral , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Retinal segmentation is a crucial step in the early warning of human health conditions. However, retinal blood vessels possess complex curvature, irregular distribution, and contain multi-scale fine structures, which make the limited receptive field of regular convolution challenging to process their vascular details efficiently. Additionally, the encoder-decoder based network leads to irreversible spatial information loss because of multiple downsampling, resulting in over-segmentation and missed segmentation of the vessels. For this reason, we develop a high-resolution network based on Deformable Convolution v3, called HRD-Net. By constructing a high-resolution representation, the network allows special attention to be paid to the details of tiny blood vessels. The proposed feature enhancement cascade module based on Deformable Convolution v3 can flexibly adapt and capture the ever-changing morphology and intricate connections of retinal blood vessels, ensuring the continuity of vessel segmentation. In the output phase of the network, the proposed global aggregation module integrates full-resolution feature maps while suppressing redundant features, achieving an effective fusion of high-level semantic information and spatial detail information. In addition, we have re-examined the selection criteria for activation and normalization methods, and also refine the network architectures from a spatial domain perspective to release redundant computational loads. Testing on the DRIVE, STARE, and CHASE_DB1 datasets indicates that HRD-Net, with fewer parameters, outperforms existing segmentation methods on several evaluation metrics such as F1, ACC, SE, SP, AUC, and IOU.