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1.
World Neurosurg ; 2024 May 30.
Artigo em Inglês | MEDLINE | ID: mdl-38823445

RESUMO

BACKGROUND: Deep brain stimulation (DBS) of the subthalamic nucleus (STN) is an effective therapy in ameliorating the motor symptoms of Parkinson's disease (PD). However, postoperative optimal contact selection is crucial for achieving the best outcome of STN-DBS surgery, but the process is currently a trial-and-error and time-consuming procedure that relies heavily on surgeons' clinical experience. METHODS: In this study, we propose a structural brain connectivity guided optimal contact selection method for STN-DBS. Firstly, we reconstruct the DBS electrode location and estimate the stimulation range using volumes of tissue activated (VTA) from each DBS contact. Then, we extract the structural connectivity features by concatenating fractional anisotropy (FA) and the number of streamlines (NOS) features of activated regions and the whole brain regions. Finally, we use a convolutional neural network (CNN) with convolutional block attention module (CBAM) to identify the structural connectivity features for the optimal contact selection. RESULTS: We review the data of 800 contacts from 100 patients with Parkinson disease for the experiment. The proposed method achieves promising results, with the average accuracy of 97.63%, average precision of 94.50%, average recall of 94.46% and average specificity of 98.18%, respectively. Our method can provide the suggestion for optimal contact selection. CONCLUSIONS: Our proposed method can improve the efficiency and accuracy of DBS optimal contact selection, reduce the dependence on surgeons' experience, and has the potential to facilitate the development of advanced DBS technology.

2.
Medicine (Baltimore) ; 103(6): e37097, 2024 Feb 09.
Artigo em Inglês | MEDLINE | ID: mdl-38335405

RESUMO

INTRODUCTION: Cellular blue nevus is an uncommon neoplasm in the spine. PATIENT CONCERNS: Here, we present a case of a 24 years old male with a 2 months history of numbness in the right upper limb and shoulder. DIAGNOSIS: Cervical spine and subcutaneous tissue invasive cellular blue nevus. INTERVENTIONS: The patient underwent C4 laminectomy and partial C3 and C5 laminectomy for total resection of the lesion. Histopathology revealed a nodular tumor with unclear boundaries, which was composed of heavily pigmented dendritic cells and more pigmented spindle cells. OUTCOMES: There was no recurrence during 3 years follow-up. CONCLUSION: Invasive cellular blue nevus of the spine can be wrongly diagnosed as spinal meningeal melanocytoma and meningeal melanoma due to its special cell behavior and rarity. Therefore, it is important to understand its pathological and clinical characteristics to avoid over-treatment.


Assuntos
Melanoma , Neoplasias Meníngeas , Nevo Azul , Nevo Pigmentado , Neoplasias Cutâneas , Humanos , Masculino , Adulto Jovem , Vértebras Cervicais/patologia , Melanoma/patologia , Neoplasias Meníngeas/patologia , Nevo Azul/diagnóstico , Nevo Azul/cirurgia , Neoplasias Cutâneas/diagnóstico , Neoplasias Cutâneas/cirurgia , Neoplasias Cutâneas/patologia
3.
Medicine (Baltimore) ; 102(43): e35654, 2023 Oct 27.
Artigo em Inglês | MEDLINE | ID: mdl-37904362

RESUMO

BACKGROUND: Rosai-Dorfman disease is a benign lymphatic tissue proliferative disease with slow growth and is rarely observed in the clinic. Most of these patients present clinically with enlarged lymph nodes. In patients with spinal extranodal Rosai-Dorfman disease, which is even rare than the disease of lymph nodes, patients may experience numbness and weakness in the extremities. CASE PRESENTATION: We report a 32-year-old male patient with multi-segmental spinal Rosai-Dorfman disease. On admission, his left fingers had been numb for 2 months. Over a 2-month period, the limb numbness progressed from the left to the right hand. The patient underwent resection of the lesion and internal fixation of the C2-C7 spine. The postoperative outcomes were satisfied and no recurrence was observed at 1-year follow-up. CONCLUSION: Spinal Rosai-Dorfman disease is a relatively rare tumor of which the pathogenesis is still unclear, and most patients have no specific clinical manifestations. Characteristic imaging findings can indicate this disease, however, a definitive diagnosis still depends on a pathological examination. Currently, total surgical resection of the tumor is a relatively effective and preferred treatment.


Assuntos
Histiocitose Sinusal , Linfadenopatia , Neoplasias , Masculino , Humanos , Adulto , Histiocitose Sinusal/diagnóstico , Histiocitose Sinusal/cirurgia , Histiocitose Sinusal/patologia , Hipestesia , Vértebras Cervicais/diagnóstico por imagem , Vértebras Cervicais/cirurgia , Vértebras Cervicais/patologia
4.
J Immunol Methods ; 521: 113550, 2023 10.
Artigo em Inglês | MEDLINE | ID: mdl-37661050

RESUMO

Black carp (Mylopharyngodon piceus) is an important fishery resource and the main breeding target in China. Due to the lack of an assay of immunoglobulin M (IgM) antibodies in black carp, there is no effective method to evaluate adaptive immune response, which limits immunological studies and vaccine development. The present study used mAbs (monoclonal antibodies) against serum IgM of grass carp as capture antibodies. The results of Western blot analysis indicated that these antibodies had strong affinity and specificity to IgM heavy chain in black carp serum and were used to detect the antibody titer, optimize the conditions, perform a sensitivity test, and develop an indirect ELISA (enzyme-linked immunosorbent assay) to detect specific IgM antibodies in the serum. This detection method has good specificity and is effective only for grass carp (Ctenopharyngodon idella) and black carp and not for crucian carp (Carassius aumtus), silver carp (Hypophthalmichthys molitrix), bighead carp (Hypophthalmichthys nobilis), mandarin fish (Siniperca chuatsi), black bream (Megalobrama skolkovii), or yellow catfish (Pseudobagrus fulvidraco). The lowest antigen detection level was 0.05 µg/ml. The error of experimental repetition in the same sample was 1.61-4.61%. The levels of specific IgM in black carp serum were steadily increased after immunization, peaked on day 28, and then slowly decreased. Indirect ELISA can be applied to detect the changes in specific antibodies in black carp serum. Moreover, indirect ELISA provides a convenient and reliable serological detection method for immunological research and evaluation of immune effects of a vaccine in black carp.


Assuntos
Carpas , Animais , Ensaio de Imunoadsorção Enzimática , Anticorpos Monoclonais , Imunoglobulina M , Imunidade Adaptativa
5.
PLoS One ; 18(4): e0284204, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37079617

RESUMO

The current evaluation of M&A performance lacks consideration of M&A motives. In this paper, we theoretically analyse and empirically test the effect of network synergy generated by M&A on the degree of realization of corporate M&A motives and the mechanism of its effect by constructing an equity network between a listed company and its subsidiaries within the company. The results show that the greater the variation of internal network node degree and strength, the more beneficial it is to promote the degree of realization of corporate M&A motivation; the results of further mechanism of action tests show that the variation of network node degree and strength has significant effects on economies of scale, economies of scope, and transaction costs; Furthermore, the heterogeneity test finds that the effect of variation of network node degree and strength to promote the degree of realization of corporate M&A motivation is more significant in the case of non-cash payment method and related M&A. This paper extends the study of complex networks to the field of M&A and uniquely explains the paradox of the "high failure rate" of M&A and the increasing activity of M&A activities from the perspective of network synergy, which helps to rationalize the M&A behavior of enterprises and further regulate the M&A behavior of listed companies by regulatory authorities.


Assuntos
Motivação , China
6.
Small Methods ; 7(6): e2201554, 2023 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-36929696

RESUMO

Metal-organic frameworks (MOFs) with inherent porosity, controllable structures, and designable components are recognized as attractive platforms for designing advanced electrodes of high-performance potassium-ion batteries (PIBs). However, the poor electrical conductivity and low theoretical capacity of many MOFs lead to inferior electrochemical performance. Herein, for the first time, a confined bismuth-organic framework with 3D porous matrix structure (Bi-MOF) as anode for PIBs via a facile wet-chemical approach is reported. Such a porous structure design with double active centers can simultaneously ensure the structure integrity and efficient charge transport to enable high-capacity electrode with super cycling life. As a result, the Bi-MOF for PIBs exhibits high reversible capacity (419 mAh g-1 at 0.1 A g-1 ), outstanding cycling stability (315 mAh g-1 at 0.5 A g-1 after 1200 cycles), and excellent full battery performance (a high energy density of 183 Wh kg-1 is achieved, outperforming all reported metal-based anodes for PIBs). Moreover, the K+ storage mechanisms of the Bi-MOF are further unveiled by in situ Raman, ex situ high-resolution transmission electron microscopy, and ex situ Fourier-transform infrared spectroscopy. This ingenious electrode design may provide further guidance for the application of MOF in energy storage systems.

7.
Protein Expr Purif ; 197: 106099, 2022 09.
Artigo em Inglês | MEDLINE | ID: mdl-35525405

RESUMO

Grass carp reovirus (GCRV) is one of the most serious pathogens threatening grass carp (Ctenopharyngon idellus) production in China. VP6 could be suitable for developing vaccine for the control of GCRV. Transgenic plants are an attractive bioreactor for their safety and ability to make economical vaccines. The B subunit of Escherichia coli heat-labile enterotoxin (LTB) fused to VP6 (LTB-VP6) was transformed into rice calli by Agrobacterium tumefaciens-mediated gene transformation. Transgenic rice calli was confirmed by PCR analysis separately. The copy numbers of LTB-VP6 inserted into the rice genome are between 1 and 2. The expression level of LTB-VP6 in rice calli was 0.0005-0.0019%, an average of 0.0011% of the TSP(total soluble proteins). LTB-VP6 was folded and assembled into a pentameric form of approximately 305 kDa capable of binding monosialoganglioside (GM1). The suitable concentration of LTB-VP6 in TSP was 0.4 µg/µl. LTB-VP6 is stable and highly active at room temperature. LTB-VP6 binding to GM1 is affected with different affinities under different temperatures. LTB-VP6 had a strong binding affinity at 25 °C and pH 8.4. Our results showed that LTB-VP6 is capable of forming an active pentameric form protein. It provides an ideal alternative to plant-based vaccines against GCRV in aquaculture.


Assuntos
Carpas , Doenças dos Peixes , Oryza , Reoviridae , Animais , Anticorpos Antivirais , Carpas/genética , Enterotoxinas/genética , Escherichia coli/metabolismo , Gangliosídeo G(M1)/metabolismo , Temperatura Alta , Oryza/genética , Reoviridae/genética
8.
PLoS One ; 16(11): e0259575, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34788332

RESUMO

Traditional forecasting methods in mergers and acquisitions (M&A) data have two limitations that significantly reduce forecasting accuracy: (1) the imbalance of data, that is, the failure cases of M&A are far fewer than the successful cases (82%/18% of our sample), and (2) both the bidder and the target of the merger have numerous descriptive features, making it difficult to choose which ones to forecast. This study proposes a neural network using partial-sigmoid (i.e., partial-sigmoid neural network [PSNN]) as the activation function of the output layer and compares three feature selection methods, namely, chi-square (chi2) test, information gain and gradient boosting decision tree (GBDT). Experimental results prove that our PSNN (improved up to 0.37 precision, 0.49 recall, 0.41 G-Mean and 0.23 F1-measure) and feature selection (improved 1.83%-13.16% accuracy) method can effectively improve the adverse effects of the defects of the above two merger data on forecasting. Scholars who studied the forecast of merger failure have overlooked three important features: assets of the previous year, market value and capital expenditure. The chi2 test feature selection method is the best among the three feature selection methods.


Assuntos
Redes Neurais de Computação , Distribuição de Qui-Quadrado
9.
ACS Appl Mater Interfaces ; 13(42): 50005-50016, 2021 Oct 27.
Artigo em Inglês | MEDLINE | ID: mdl-34637269

RESUMO

Practical applications of carbon anodes in high-power potassium-ion batteries (PIBs) were hampered by their limited rate properties, due to the sluggish K+ transport kinetics in the bulk. Constructing convenient ion/electron transfer channels in the electrode is of great importance to realize fast charge/discharge rates. Here, cross-linked porous carbon nanofibers (inner porous carbon nanotubes and outer soft carbon layer) modified with oxygen-containing functional groups were well designed as anodes to realize robust de-/potassiation kinetics. The novel anode delivered excellent rate capabilities (107 mAh g-1 at 20 A g-1 and 78 mAh g-1 at 40 A g-1) and superior cycling stability (76% capacity retention after 14,000 cycles at 2 A g-1). In situ XRD measurement, in situ Raman spectra, and galvanostatic intermittent titration verified its surface-dominated potassium storage behavior with fast de-/potassiation kinetics, excellent reversibility, and rapid ion/electron transport. Moreover, theoretical investigation revealed that the carboxyl groups in the carbon offered additional capacitive adsorption sites for K+, thus significantly enhancing the reversible capacity. Surprisingly, a full cell using the anode and perylene-3,4,9,10-tetracarboxylic dianhydride cathode achieved an outstanding power density of 23,750 W kg-1 and superior fast charge/slow discharge performance.

10.
Eur Radiol ; 31(12): 9324-9334, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34109485

RESUMO

OBJECTIVES: Mild cognitive impairment (MCI) is a well-defined non-motor manifestation and a harbinger of dementia in Parkinson's disease. This study is to investigate brain connectivity markers of MCI using diffusion tensor imaging and resting-state functional MRI, and help MCI diagnosis in PD patients. METHODS: We evaluated 131 advanced PD patients (disease duration > 5 years; 59 patients with MCI) and 48 healthy control subjects who underwent a diffusion-weighted and resting-state functional MRI scanning. The patients were randomly assigned to training (n = 100) and testing (n = 31) groups. According to the Brainnetome Atlas, ROI-based structural and functional connectivity analysis was employed to extract connectivity features. To identify features with significant discriminative power for patient classification, all features were put into an all-relevant feature selection procedure within cross-validation loops. RESULTS: Nine features were identified to be significantly relevant to patient classification. They showed significant differences between PD patients with and without MCI and positively correlated with the MoCA score. Five of them did not differ between general MCI subjects and healthy controls from the ADNI database, which suggested that they could uniquely play a part in the MCI diagnosis of PD. On basis of these relevant features, the random forest model constructed from the training group achieved an accuracy of 83.9% in the testing group, to discriminate patients with and without MCI. CONCLUSIONS: The results of our study provide preliminary evidence that structural and functional connectivity abnormalities may contribute to cognitive impairment and allow to predict the outcome of MCI diagnosis in PD. KEY POINTS: • Nine MCI markers were identified using an all-relevant feature selection procedure. • Five of nine markers differed between MCI and NC in PD, but not in general persons. • A random forest model achieved an accuracy of 83.9% for MCI diagnosis in PD.


Assuntos
Disfunção Cognitiva , Doença de Parkinson , Encéfalo/diagnóstico por imagem , Disfunção Cognitiva/diagnóstico por imagem , Imagem de Tensor de Difusão , Humanos , Imageamento por Ressonância Magnética , Testes Neuropsicológicos , Doença de Parkinson/complicações , Doença de Parkinson/diagnóstico por imagem
11.
Biomed Res Int ; 2021: 3642463, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33542919

RESUMO

Type 2 diabetes mellitus (T2DM) is a common chronic metabolic disease. Accumulating evidence has demonstrated that nonalcoholic fatty liver disease (NAFLD) shares common typical features with T2DM, and they affect each other extensively. Thus, NAFLD has emerged as a novel target for T2DM prevention and care. Although Corni Fructus (CF) and its extracts have a therapeutic effect on T2DM, its effects and mechanisms on T2DM with NAFLD are far from elucidated. In this study, a mouse model of T2DM with NAFLD complication was established in ICR mice by feeding a high-fat, high-sugar (HFHS) diet and intraperitoneally injecting with a low dose of streptozotocin (STZ). Then, the effects of iridoid glycosides (IG) extracted from CF on this mouse model were investigated. We found that 4-week IG administration remarkably alleviated hyperglycemia and insulin resistance and significantly reduced inflammation, oxidative stress, and fat accumulation in the liver of T2DM with NAFLD mice. Further studies showed that IG inhibited the NF-κB but enhanced the PI3K-AKT signaling pathway. In summary, these results indicated that the IG from CF has potential therapeutic effects on T2DM with NAFLD.


Assuntos
Diabetes Mellitus Experimental/tratamento farmacológico , Diabetes Mellitus Experimental/metabolismo , Inflamação/prevenção & controle , Glicosídeos Iridoides/farmacologia , Hepatopatia Gordurosa não Alcoólica/tratamento farmacológico , Hepatopatia Gordurosa não Alcoólica/metabolismo , Animais , Cornus/química , Diabetes Mellitus Experimental/patologia , Dieta Hiperlipídica , Modelos Animais de Doenças , Inflamação/etiologia , Inflamação/patologia , Masculino , Camundongos , Camundongos Endogâmicos ICR , Hepatopatia Gordurosa não Alcoólica/patologia , Estresse Oxidativo/fisiologia , Substâncias Protetoras/farmacologia , Transdução de Sinais
12.
Nanoscale Horiz ; 5(12): 1643, 2020 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-33118586

RESUMO

Correction for 'Nitrogen-doped carbon nanotubes as an anode for a highly robust potassium-ion hybrid capacitor' by Xiuqi Li et al., Nanoscale Horiz., 2020, DOI: .

13.
Nanoscale Horiz ; 5(12): 1586-1595, 2020 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-33052993

RESUMO

Potassium ion hybrid capacitors (KIHCs) have drawn growing interest owing to their outstanding energy density, power density and excellent cycling stability. However, the large ionic radius of potassium triggers a huge volume change during continuous K+ insertion/extraction processes, restricting the development of KIHCs. Here, we report N-doped carbon nanotubes (NCNTs) for high-performance K+ storage. The NCNTs possess a hierarchical structure and N functional groups and not only offer sufficient space to relieve the volume expansion, but also provide highly efficient channels to transport electrons and ions. As a result, the NCNTs anode presents a high specific capacity and an excellent cycling stability with an average decay rate of 0.0238% per cycle (the lowest value among the reported carbon-based anodes for K-ions batteries) during 3600 continuous cycles. A potassium ion hybrid capacitor (KIHC) was also designed with the NCNT anode and a commercial active carbon cathode and achieved both a high energy/power density (117.1 W h kg-1/1713.4 W kg-1) and a long cycle life (2000 cycles at 1 A g-1). Moreover, the in situ Raman and ex situ element mapping characterization demonstrate the outstanding electrochemical reversibility of the NCNTs. This work provides a superior strategy to design low-cost anode materials with excellent K+ storage electrochemistry.

14.
Am J Chin Med ; 48(6): 1385-1407, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32907359

RESUMO

Accumulating evidence suggests that gut microbiota plays a crucial role in the development of metabolic diseases, especially type 2 diabetes mellitus (T2DM). The nutrient-rich resource Cornus Fructus (CF) showed curative effects on diabetes mellitus. However, the mechanism underlying its hyperglycemic activity remains obscure. Herein, the antidiabetic potential of four extracts from CF, including saponin (CTS), iridoid glycoside (CIG), tannin (CT), and alcohol extract (CCA) was evaluated in vivo. The results showed that all four extracts could increase the body weight, decrease the blood glucose levels, and elevate the glucose tolerance. Moreover, insulin sensitivity and lipid profile were significantly improved in fed mice. In the [Formula: see text]-diversity index of samples, compared to the DM group, the diversity and richness of gut microbiota in mice to a certain extent were reduced in both CF extracts and Metformin (PC). Among them, there was statistical significance in PC (ACE, [Formula: see text]) and CCA (ACE, [Formula: see text]; chao1: [Formula: see text]). Beta diversity showed the same trend as the UPGMA clustering trees, which revealed that CF extracts could improve intestinal homeostasis in T2DM mice. Also, CF extracts could elevate the production of short-chain fatty acids, as well as regulate the composition of gut microbiota. The key bacteria related to T2DM including Firmicutes, Bacteroides, Lactobacillus, and Clostridium were modulated by metformin and CF. Altogether, CF is a potential nutrient-rich candidate that can be used in functional foods for the treatment of T2DM, and the change of gut microbiota might be a novel mechanism underlying its hyperglycemic activity.


Assuntos
Cornus/química , Diabetes Mellitus Tipo 2/tratamento farmacológico , Diabetes Mellitus Tipo 2/microbiologia , Microbioma Gastrointestinal/efeitos dos fármacos , Fitoterapia , Extratos Vegetais/farmacologia , Extratos Vegetais/uso terapêutico , Animais , Glicemia/metabolismo , Peso Corporal/efeitos dos fármacos , Diabetes Mellitus Tipo 2/etiologia , Modelos Animais de Doenças , Alimento Funcional , Resistência à Insulina , Glicosídeos Iridoides , Metabolismo dos Lipídeos/efeitos dos fármacos , Masculino , Camundongos Endogâmicos ICR , Extratos Vegetais/química , Saponinas , Taninos
15.
iScience ; 23(3): 100919, 2020 Mar 27.
Artigo em Inglês | MEDLINE | ID: mdl-32114378

RESUMO

Lithium selenium (Li-Se) batteries have attracted increasing interest for its high theoretical volumetric capacities up to 3,253 Ah L-1. However, current studies are largely limited to electrodes with rather low mass loading and low areal capacity, resulting in low volumetric performance. Herein, we report a design of covalent selenium embedded in hierarchical nitrogen-doped carbon nanofibers (CSe@HNCNFs) for ultra-high areal capacity Li-Se batteries. The CSe@HNCNFs provide excellent ion and electron transport performance, whereas effectively retard polyselenides diffusion during cycling. We show that the Li-Se battery with mass loading of 1.87 mg cm-2 displays a specific capacity of 762 mAh g-1 after 2,500 cycles, with almost no capacity fading. Furthermore, by increasing the mass loading to 37.31 mg cm-2, ultra-high areal capacities of 7.30 mAh cm-2 is achieved, which greatly exceeds those reported previously for Li-Se batteries.

16.
Int J Biol Macromol ; 151: 814-820, 2020 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-32097736

RESUMO

Grass carp reovirus (GCRV) is one of the most serious pathogens threatening grass carp (Ctenopharyngodon idella) production and results in high mortality in China. To obtain a genetically engineered oral vaccine against GCRV, the cholera toxin B subunit (CTB) of Vibrio cholerae was fused to VP7 (CTB-VP7) and transformed into BL21(DE3) for expression. SDS-PAGE and Western blotting showed that the purified rCTB-VP7 fusion protein (rCTB-VP7) was approximately 49.0 kDa. The monomeric nature of rCTB-VP7 through multistage purification showed a binding affinity for GM1, a receptor for biologically active CTB. rCTB-VP7 is not vulnerable to disassembly by SDS but is vulnerable to disassembly by 2-mercaptoethanol. rCTB-VP7 is stable and highly active at room temperature. The binding affinity experiment between rCTB-VP7 and GM1 also confirms the effects of acid and alkalinity in solution on the structure of rCTB-VP7. rCTB-VP7 bound to GM1 with different affinities under different temperatures and pH values. Prokaryotic expression of rCTB-VP7 was characterized by high expression and easy purification and had a strong binding force with GM1 at 37 °C and pH 7.4. Our results suggest that rCTB-VP7 has the potential as an oral vaccine for protection against GCRV in aquaculture.


Assuntos
Antígenos Virais/química , Antígenos Virais/genética , Proteínas do Capsídeo/química , Proteínas do Capsídeo/genética , Toxina da Cólera/genética , Expressão Gênica , Dobramento de Proteína , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/genética , Antígenos Virais/isolamento & purificação , Proteínas do Capsídeo/isolamento & purificação , Fenômenos Químicos , Escherichia coli/genética , Concentração de Íons de Hidrogênio , Conformação Proteica , Proteínas Recombinantes de Fusão/isolamento & purificação , Relação Estrutura-Atividade , Temperatura
17.
Fish Shellfish Immunol ; 98: 766-772, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31734284

RESUMO

Infectious hypodermal and haematopoietic necrosis virus (IHHNV) is a major viral pathogen in cultured penaeid shrimp. IHHNV has many hosts, mainly including crustaceans. It has recently been reported that Procambarus clarkii can be infected by IHHNV. In the present study, we studied the hepatopancreas of P. clarkii by transcriptome high-throughput sequencing to analyze the response of P. clarkii to IHHNV infection. After de novo assembly, there were 400,340,760 clean reads. A total of 237 differentially expressed genes (DEGs) were obtained, including 77 significantly up-regulated unigenes and 160 significantly down-regulated ones. The expression levels of 12 immune-related DEGs were validated by qRT-PCR, substantiating the reliability of RNA-Seq results. The enrichment analysis of DEGs showed that the immune-related pathways were closely related to apoptosis and phagocytosis. Moreover, a large number of pathways related to metabolic function were down-regulated, suggesting that IHHNV infection might affect the growth of P. clarkii.


Assuntos
Proteínas de Artrópodes/metabolismo , Astacoidea/imunologia , Densovirinae/fisiologia , Regulação da Expressão Gênica , Hepatopâncreas/virologia , Transcriptoma , Animais , Astacoidea/virologia , Perfilação da Expressão Gênica , Hepatopâncreas/imunologia , Sequenciamento de Nucleotídeos em Larga Escala
18.
Protein Expr Purif ; 158: 1-8, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-30753891

RESUMO

Grass carp reovirus (GCRV) is one of the most serious pathogens threatening grass carp (Ctenopharyngodon idellus) production and results in high mortality in China. VP7 from GCRV is involved in viral infection and could be suitable for developing vaccines for the control of GCRV infection. To obtain a genetically engineered vaccine and a plant-based oral vaccine and to evaluate their immune efficacy as an oral vaccine against GCRV, cholera toxin B subunit (CTB) of Vibrio cholerae fused to VP7 (CTB-VP7) was transformed into BL21(DE3) for expression. SDS-PAGE and Western blotting showed that the purified CTB-VP7 fusion protein (rCTB-VP7) was approximately 49.0 kDa. Meanwhile, CTB-VP7 was transformed into rice callus cells by Agrobacterium tumefaciens-mediated gene transformation. CTB-VP7 was integrated into the nuclear genome by PCR, and mRNA transcripts of CTB-VP7 were detected. ELISA and Western blot analyses revealed that the CTB-VP7 fusion protein (CTB-VP7) could be expressed in rice callus lines. The level of expression was determined to be 1.54% ±â€¯0.43 of the total soluble protein. CTB-VP7 showed a binding affinity for monosialoganglioside(GM1), a receptor for CTB. CTB-VP7 showed a higher affinity towards GM1 compared to rCTB-VP7. CTB-VP7 bonded to GM1 with different affinities under different temperatures. Maximum binding of CTB-VP7 to GM1 was reported to occur within 2 h at 37 °C, and approximately half of the binding affinity remained at 25 °C. Our results suggest that CTB-VP7 could be produced in rice calli, increasing the possibility that edible plants can be employed in mucosal vaccines for protection against GCRV in aquaculture.


Assuntos
Antígenos Virais/imunologia , Carpas/imunologia , Toxina da Cólera , Doenças dos Peixes/prevenção & controle , Infecções por Reoviridae/prevenção & controle , Reoviridae/imunologia , Vacinas Virais/imunologia , Animais , Antígenos Virais/química , Antígenos Virais/genética , Carpas/virologia , Toxina da Cólera/química , Toxina da Cólera/genética , Toxina da Cólera/imunologia , Toxina da Cólera/isolamento & purificação , Doenças dos Peixes/genética , Doenças dos Peixes/imunologia , Oryza/química , Oryza/genética , Oryza/imunologia , Plantas Geneticamente Modificadas/química , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/imunologia , Proteínas Recombinantes de Fusão , Reoviridae/genética , Infecções por Reoviridae/genética , Infecções por Reoviridae/imunologia , Infecções por Reoviridae/veterinária , Vacinas Virais/química , Vacinas Virais/genética
19.
Cell Physiol Biochem ; 48(3): 1088-1098, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30041238

RESUMO

BACKGROUND/AIMS: To investigate the mechanism that enables oxidative stress and cytoskeleton protein carbonylation to contribute to axonal dysfunction in traumatic brain injury (TBI). METHODS: We created an in vitro model of neuronal oxidative damage by exposing a neuron-like cell line (PC-12) to different concentrations (100 µM, 200 µM, and 300 µM) of H2O2 for 24 h or 48 h. Carbonyl modification of cytoskeletal proteins (ß-actin and ß-tubulin) and its impact on ß-actin/ß-tubulin filament dynamics were determined by enzyme-linked immunosorbent assay, immunostaining, and western blotting. Depolymerization of ß-actin/ß-tubulin filaments was evaluated using the monomer/polymer ratio of each protein via western blotting. Phosphorylation of the neurofilament heavy chain (P-NFH) was used as an axonal injury marker and detected by immunostaining. RESULTS: Our results showed that H2O2 treatment led to increased oxidative stress in PC-12 cells, as indicated by the increased generation of malondialdehyde and 8-hydroxydeoxyguanosine and decreased intracellular glutathione levels. H2O2 treatment also increased carbonyl modification of total proteins and cytoskeleton proteins ß-actin/ß-tubulin, which occurred concurrently with the suppression of proteasome activity. Moreover, H2O2 treatment increased the generation of the axonal injury marker P-NFH, and depolymerization of the ß-actin/ß-tubulin filaments was indicated by increased monomer/polymer ratios of each protein. Lastly, overexpression of the proteasome ß5 subunit in PC-12 cells significantly reduced the H2O2-induced accumulation of carbonylated ß-actin/ ß-tubulin, P-NFH, and ß-actin/ß-tubulin depolymerization. CONCLUSIONS: We concluded that carbonylation of cytoskeleton proteins could lead to depolymerization of their filaments and axonal injury, and proteasome suppression contributes to the accumulation of carbonylated proteins under oxidative conditions.


Assuntos
Citoesqueleto de Actina/efeitos dos fármacos , Proteínas do Citoesqueleto/metabolismo , Peróxido de Hidrogênio/farmacologia , Carbonilação Proteica/efeitos dos fármacos , 8-Hidroxi-2'-Desoxiguanosina , Citoesqueleto de Actina/metabolismo , Animais , Desoxiguanosina/análogos & derivados , Desoxiguanosina/metabolismo , Glutationa/metabolismo , Malondialdeído/metabolismo , Proteínas de Neurofilamentos/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Células PC12 , Fosforilação/efeitos dos fármacos , Complexo de Endopeptidases do Proteassoma/genética , Complexo de Endopeptidases do Proteassoma/metabolismo , Subunidades Proteicas/genética , Subunidades Proteicas/metabolismo , Ratos , Tubulina (Proteína)/metabolismo
20.
Exp Ther Med ; 14(1): 344-348, 2017 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-28672936

RESUMO

The purpose of this study was to further evaluate the role of myxoma virus (MYXV) as an oncolytic agent against experimental human gliomas in vitro, and analyze the effect of MYXV on malignant glioma cells at different incubation periods and infected at different multiplicities of infection. Neuroglioma cell lines U251 and A172 were cultured with various infective doses of myxoma virus at different time points (0-3 days) and cellular survival rates were evaluated using an MTT assay. Cell viability and cell death rates were assessed using Annexin V/propidium iodide and applying flow cytometry. Furthermore, the expression levels of phosphorylated AKT (p-AKT) in malignant gliomas were detected by western blot analysis to investigate the possible cell signaling targets in the pathway. MYXV exhibited a dose and time-dependent cytotoxic effect on neuroglioma cells, and there was increased expression of p-AKT in malignant gliomas. The present study confirms that MYXV induces oncolysis of malignant gliomas through regulating the activation of AKT. As such, MYXV is a potential therapeutic agent against human malignant gliomas.

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