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1.
Cerebrovasc Dis ; 52(3): 318-343, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36279857

RESUMO

BACKGROUND: There is increasing evidence on the prognostic significance of D-dimer and fibrinolysis in stroke. However, the systematic analysis of their relationship with adverse outcomes after stroke is lacking. Herein, we comprehensively assessed the correlation of D-dimer and fibrinolysis with stroke outcomes through meta-analysis. METHODS: Studies for systematic literature review were retrieved from PubMed, EMBASE, and Cochrane Library databases. The association of D-dimer and fibrinolysis with outcomes of stroke patients was expressed as an odds ratio (OR) with 95% confidence intervals (95% CI). RESULTS: Totally, 52 studies comprising 21,473 stroke patients were included. The results showed that the high D-dimer level was significantly associated with peripheral venous thrombosis after stroke (OR 1.03, 95% CI 1.01-1.05), poor outcome (MRS >2) after stroke (OR 1.731, 95% CI 1.464-2.048), death after stroke (OR 2.367, 95% CI 1.737-3.224), stroke recurrence (OR 1.229, 95% CI 1.113-1.358), and early neurologic deterioration (NIHSS >4) (OR 1.791, 95% CI 1.117-2.870). Moreover, high fibrinogen level was significantly associated with poor outcome (MRS >2) after stroke (OR 1.650, 95% CI 1.314-2.071), death after stroke (OR 1.310, 95% CI 1.128-1.520), stroke recurrence (OR 1.228, 95% CI 1.166-1.422), early neurologic deterioration (NIHSS >4) (OR 2.381, 95% CI 1.156-4.904), and coronary events after stroke (OR 1.427, 95% CI 1.232-1.653). CONCLUSION: Fibrinogen and D-dimer may be associated with adverse outcomes in patients with stroke, suggesting that they may serve as possible biomarkers for post-stroke adverse outcomes.


Assuntos
Hemostáticos , Acidente Vascular Cerebral , Humanos , Fibrinogênio , Relevância Clínica , Produtos de Degradação da Fibrina e do Fibrinogênio/análise , Acidente Vascular Cerebral/diagnóstico , Acidente Vascular Cerebral/terapia , Biomarcadores
2.
Curr Genomics ; 24(3): 155-170, 2023 Nov 22.
Artigo em Inglês | MEDLINE | ID: mdl-38178986

RESUMO

Background: Recent studies on CRISPR/Cas9-mediated gene editing in Schistosoma mansoni have shed new light on the study and control of this parasitic helminth. However, the gene editing efficiency in this parasite is modest. Methods: To improve the efficiency of CRISPR/Cas9 genome editing in schistosomes, we used lentivirus, which has been effectively used for gene editing in mammalian cells, to deliver plasmid DNA encoding Cas9 nuclease, a sgRNA targeting acetylcholinesterase (SmAChE) and a mCherry fluorescence marker into schistosomes. Results: MCherry fluorescence was observed in transduced eggs, schistosomula, and adult worms, indicating that the CRISPR components had been delivered into these parasite stages by lentivirus. In addition, clearly changed phenotypes were observed in SmAChE-edited parasites, including decreased SmAChE activity, reduced hatching ability of edited eggs, and altered behavior of miracidia hatched from edited eggs. Next-generation sequencing analysis demonstrated that the lentiviral transduction-based CRISPR/Cas9 gene modifications in SmAChE-edited schistosomes were homology-directed repair predominant but with much lower efficiency than that obtained using electroporation (data previously published by our laboratory) for the delivery of CRISPR components. Conclusion: Taken together, electroporation is more efficient than lentiviral transduction in the delivery of CRISPR/Cas9 into schistosomes for programmed genome editing. The exploration of tactics for enhancing CRISPR/Cas9 gene editing provides the basis for the future improvement of programmed genome editing in S. mansoni.

3.
Bioessays ; 43(1): e2000185, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-33145822

RESUMO

Recent reports of CRISPR/Cas9 genome editing in parasitic helminths open up new avenues for research on these dangerous pathogens. However, the complex morphology and life cycles inherent to these parasites present obstacles for the efficient application of CRISPR/Cas9-targeted mutagenesis. This is especially true with the trematode flukes where only modest levels of gene mutation efficiency have been achieved. Current major challenges in the application of CRISPR/Cas9 for study of parasitic worms thus lie in enhancing gene mutation efficiency and overcoming issues involved in host passage so that mutated parasites survive. Strategies developed for CRISPR/Cas9 studies on Caenorhabditis elegans, protozoa and mammalian cells, including novel delivery methods, the choice of selectable markers, and refining mutation precision represent novel tactics whereby these impediments can be overcome. Furthermore, employing CRISPR/Cas9-mediated gene drive to interfere with vector transmission represents a novel approach for the control of parasitic worms that is worthy of further exploration.


Assuntos
Sistemas CRISPR-Cas , Parasitos , Animais , Sistemas CRISPR-Cas/genética , Caenorhabditis elegans/genética , Edição de Genes , Mutagênese
4.
FASEB J ; 35(1): e21205, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-33337558

RESUMO

CRISPR/Cas9-mediated genome editing shows cogent potential for the genetic modification of helminth parasites. We report successful gene knock-in (KI) into the genome of the egg of Schistosoma mansoni by combining CRISPR/Cas9 with single-stranded oligodeoxynucleotides (ssODNs). We edited the acetylcholinesterase (AChE) gene of S. mansoni targeting two guide RNAs (gRNAs), X5 and X7, located on exon 5 and exon 7 of Smp_154600, respectively. Eggs recovered from livers of experimentally infected mice were transfected by electroporation with a CRISPR/Cas9-vector encoding gRNA X5 or X7 combining with/ without a ssODN donor. Next generation sequencing analysis of reads of amplicon libraries spanning targeted regions revealed that the major modifications induced by CRISPR/Cas9 in the eggs were generated by homology directed repair (HDR). Furthermore, soluble egg antigen from AChE-edited eggs exhibited markedly reduced AChE activity, indicative that programed Cas9 cleavage mutated the AChE gene. Following injection of AChE-edited schistosome eggs into the tail veins of mice, an significantly enhanced Th2 response involving IL-4, -5, -10, and-13 was detected in lung cells and splenocytes in mice injected with X5-KI eggs in comparison to control mice injected with unmutated eggs. A Th2-predominant response, with increased levels of IL-4, -13, and GATA3, also was induced by X5 KI eggs in small intestine-draining mesenteric lymph node cells when the gene-edited eggs were introduced into the subserosa of the ileum of the mice. These findings confirmed the potential and the utility of CRISPR/Cas9-mediated genome editing for functional genomics in schistosomes.


Assuntos
Acetilcolinesterase/metabolismo , Sistemas CRISPR-Cas , Edição de Genes , Proteínas de Helminto/metabolismo , Schistosoma mansoni/enzimologia , Esquistossomose mansoni/metabolismo , Acetilcolinesterase/genética , Animais , Feminino , Proteínas de Helminto/genética , Camundongos , Schistosoma mansoni/genética , Esquistossomose mansoni/genética
5.
Environ Res ; 213: 113633, 2022 10.
Artigo em Inglês | MEDLINE | ID: mdl-35700766

RESUMO

The bentonite colloid produced in the deep geological repository of high-level radioactive waste can directly affect the migration of radionuclide strontium when it acts on claystone. The adsorption characteristics of strontium were investigated on claystone with the presence or absence of bentonite colloids from the Suhongtu area of China. The effects of different influencing factors, such as pH and solid content, on the adsorption process were investigated by batch adsorption experiments, and spectroscopic techniques were used to characterize the samples before and after adsorption of strontium. The results show that the presence of bentonite colloids can promote the adsorption of strontium on claystone under alkaline conditions. and the general order kinetic model provided the best fit to the experimental data. Strontium is adsorbed on the surface of claystone and bentonite colloid by ion exchange and surface complexation. Most of the Sr2+ formed SrCO3 with CO32- after ion exchange with Ca2+ and Mg2+ in plagioclase and dolomite, and a small amount of Sr2+ was adsorbed by complexation with -OH, Al-O and Si-O. These results provide a scientific basis for predicting the migration of strontium in subsurface porous media and the siting of high-level radioactive waste repositories.


Assuntos
Resíduos Radioativos , Adsorção , Bentonita/química , Coloides/química , Resíduos Radioativos/análise , Estrôncio/química
6.
BMC Med Imaging ; 21(1): 195, 2021 12 24.
Artigo em Inglês | MEDLINE | ID: mdl-34952572

RESUMO

BACKGROUND: Magnetic resonance imaging (MRI) is an effective auxiliary diagnostic method in clinical medicine, but it has always suffered from the problem of long acquisition time. Compressed sensing and parallel imaging are two common techniques to accelerate MRI reconstruction. Recently, deep learning provides a new direction for MRI, while most of them require a large number of data pairs for training. However, there are many scenarios where fully sampled k-space data cannot be obtained, which will seriously hinder the application of supervised learning. Therefore, deep learning without fully sampled data is indispensable. MAIN TEXT: In this review, we first introduce the forward model of MRI as a classic inverse problem, and briefly discuss the connection of traditional iterative methods to deep learning. Next, we will explain how to train reconstruction network without fully sampled data from the perspective of obtaining prior information. CONCLUSION: Although the reviewed methods are used for MRI reconstruction, they can also be extended to other areas where ground-truth is not available. Furthermore, we may anticipate that the combination of traditional methods and deep learning will produce better reconstruction results.


Assuntos
Aprendizado Profundo , Processamento de Imagem Assistida por Computador/métodos , Imageamento por Ressonância Magnética , Humanos
7.
Sensors (Basel) ; 20(8)2020 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-32326501

RESUMO

Resource constraints have prevented comprehensive cryptography and multifactor authentication in numerous Internet of Things (IoT) connectivity scenarios. Existing IoT systems generally adopt lightweight security protocols that lead to compromise and privacy leakage. Edge computing enables better access control and privacy protection, furthermore, blockchain architecture has achieved a trusted store of value by open-source and distributed consensus mechanisms. To embrace these new paradigms, we propose a scheme that employs one-time association multitasking proofs for peer to local authentication (OTMP-P2L). The scheme chooses relevant nondeterministic polynomial (NP) problem tasks, and manages localized trust and anonymity by using smart devices such as phones and pads, thereby enabling IoT devices to autonomously perform consensus validation with an enhanced message authentication code. This nested code is a one-time zero-knowledge proof that comprises multiple logic verification arguments. To increase diversity and reduce the workload of each one, these arguments are chained by a method that establishes some of the inputs of the following task from the output of previous tasks. We implemented a smart lock system and confirmed that the scheme outperforms IoT authentication methods. The result demonstrates superior flexibility through dynamic difficulty strategies and succinct non-interactive peer-to-peer (P2P) verification.

8.
Int J Mol Sci ; 21(11)2020 Jun 04.
Artigo em Inglês | MEDLINE | ID: mdl-32512920

RESUMO

We characterized Schistosoma japonicum HSP40 (Sjp40) and HSP90α (Sjp90α) in this study. Western blot analysis revealed both are present in soluble egg antigens and egg secretory proteins, implicating them in triggering the host immune response after secretion from eggs into host tissues. These observations were confirmed by immunolocalization showing both HSPs are located in the Reynolds' layer within mature eggs, suggesting they are secreted by miracidia and accumulate between the envelope and the eggshell. Both HSPs are present in the musculature and parenchyma of adult males and in the vitelline cells of females; only Sjp90α is present on the tegument of adults. Sjp40 was able to enhance the expression of macrophages, dendritic cells, and eosinophilic cells in mouse liver non-parenchymal cells, whereas rSjp90α only stimulated the expression of dendritic cells. T helper 1 (Th1), Th2, and Th17 responses were increased upon rSjp40 stimulation in vitro, but rSjp90 only stimulated an increased Th17 response. Sjp40 has an important role in reducing the expression of fibrogenic gene markers in hepatic stellate cells in vitro. Overall, these findings provide new information on HSPs in S. japonicum, improving our understanding of the pathological roles they play in their interaction with host immune cells.


Assuntos
Antígenos de Helmintos/imunologia , Proteínas de Choque Térmico HSP40/imunologia , Proteínas de Choque Térmico HSP90/imunologia , Proteínas de Helminto/imunologia , Schistosoma japonicum/imunologia , Sequência de Aminoácidos , Animais , Antígenos de Helmintos/química , Antígenos de Helmintos/metabolismo , Modelos Animais de Doenças , Proteínas de Choque Térmico HSP40/química , Proteínas de Choque Térmico HSP40/metabolismo , Proteínas de Choque Térmico HSP90/química , Proteínas de Choque Térmico HSP90/metabolismo , Proteínas de Helminto/química , Proteínas de Helminto/metabolismo , Células Estreladas do Fígado/metabolismo , Imuno-Histoquímica , Fígado/imunologia , Fígado/metabolismo , Fígado/parasitologia , Fígado/patologia , Camundongos , Modelos Moleculares , Conformação Proteica , Schistosoma japonicum/metabolismo , Relação Estrutura-Atividade
9.
Int J Mol Sci ; 20(7)2019 Mar 28.
Artigo em Inglês | MEDLINE | ID: mdl-30925781

RESUMO

We showed previously that the Schistosoma japonicum insulin-like peptide (SjILP) binds the worm insulin receptors, thereby, activating the parasite's insulin pathway and emphasizing its important role in regulating uptake of glucose, a nutrient essential for parasite survival. Here we show that SjILP is differentially expressed in the schistosome life cycle and is especially highly transcribed in eggs, miracidia, and adult female worms. RNA inference was employed to knockdown SjILP in adults in vitro, with suppression confirmed by significantly reduced protein production, declined adenosine diphosphate levels, and reduction in glucose consumption. Immunolocalization showed that SjILP is located to lateral gland cells of mature intra-ovular miracidia in the schistosome egg, and is distributed on the ciliated epithelium and internal cell masses of newly transformed miracidia. In schistosomula, SjILP is present on the tegument in two antero-lateral points, indicating highly polarized expression during cercarial transformation. Analysis of serum from S. japonicum-infected mice by ELISA using a recombinant form of SjILP as an antigen revealed IgG immunoreactivity to this molecule at 7 weeks post-infection indicating it is likely secreted from mature eggs into the host circulation. These findings provide further insights on ILP function in schistosomes and its essential roles in parasite survival and growth in different development stages.


Assuntos
Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Helminto/genética , Insulina/genética , Schistosoma japonicum/crescimento & desenvolvimento , Schistosoma japonicum/genética , Esquistossomose Japônica/parasitologia , Animais , Feminino , Proteínas de Helminto/imunologia , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Insulina/imunologia , Estágios do Ciclo de Vida , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Schistosoma japonicum/imunologia , Esquistossomose Japônica/sangue , Esquistossomose Japônica/imunologia
10.
BMC Med Imaging ; 18(1): 7, 2018 05 03.
Artigo em Inglês | MEDLINE | ID: mdl-29724180

RESUMO

BACKGROUND: Multi-contrast images in magnetic resonance imaging (MRI) provide abundant contrast information reflecting the characteristics of the internal tissues of human bodies, and thus have been widely utilized in clinical diagnosis. However, long acquisition time limits the application of multi-contrast MRI. One efficient way to accelerate data acquisition is to under-sample the k-space data and then reconstruct images with sparsity constraint. However, images are compromised at high acceleration factor if images are reconstructed individually. We aim to improve the images with a jointly sparse reconstruction and Graph-based redundant wavelet transform (GBRWT). METHODS: First, a sparsifying transform, GBRWT, is trained to reflect the similarity of tissue structures in multi-contrast images. Second, joint multi-contrast image reconstruction is formulated as a ℓ2, 1 norm optimization problem under GBRWT representations. Third, the optimization problem is numerically solved using a derived alternating direction method. RESULTS: Experimental results in synthetic and in vivo MRI data demonstrate that the proposed joint reconstruction method can achieve lower reconstruction errors and better preserve image structures than the compared joint reconstruction methods. Besides, the proposed method outperforms single image reconstruction with joint sparsity constraint of multi-contrast images. CONCLUSIONS: The proposed method explores the joint sparsity of multi-contrast MRI images under graph-based redundant wavelet transform and realizes joint sparse reconstruction of multi-contrast images. Experiment demonstrate that the proposed method outperforms the compared joint reconstruction methods as well as individual reconstructions. With this high quality image reconstruction method, it is possible to achieve the high acceleration factors by exploring the complementary information provided by multi-contrast MRI.


Assuntos
Encéfalo/diagnóstico por imagem , Processamento de Imagem Assistida por Computador/métodos , Imageamento por Ressonância Magnética/métodos , Meios de Contraste , Humanos , Processamento de Sinais Assistido por Computador , Análise de Ondaletas
11.
Sensors (Basel) ; 18(3)2018 Mar 06.
Artigo em Inglês | MEDLINE | ID: mdl-29509666

RESUMO

Deep convolutional neural networks (CNNs) are successful in single-image super-resolution. Traditional CNNs are limited to exploit multi-scale contextual information for image reconstruction due to the fixed convolutional kernel in their building modules. To restore various scales of image details, we enhance the multi-scale inference capability of CNNs by introducing competition among multi-scale convolutional filters, and build up a shallow network under limited computational resources. The proposed network has the following two advantages: (1) the multi-scale convolutional kernel provides the multi-context for image super-resolution, and (2) the maximum competitive strategy adaptively chooses the optimal scale of information for image reconstruction. Our experimental results on image super-resolution show that the performance of the proposed network outperforms the state-of-the-art methods.

12.
Int J Mol Sci ; 19(10)2018 Oct 09.
Artigo em Inglês | MEDLINE | ID: mdl-30304851

RESUMO

There is a pressing need to develop vaccines for schistosomiasis given the current heavy dependency on praziquantel as the only available drug for treatment. We previously showed the ligand domain of the Schistosoma japonicum insulin receptor 1 and 2 (rSjLD1 and 2) fusion proteins conferred solid protection in mice against challenge infection with S. japonicum. To improve vaccine efficacy, we compared the immunogenicity and protective efficacy of rSjLD1 on its own and in combination with S. japonicum triose-phosphate isomerase (SjTPI), formulated with either of two adjuvants (QuilA and montanide ISA 720VG) in murine vaccine trials against S. japonicum challenge. The level of protection was higher in mice vaccinated only with rSjLD1 formulated with either adjuvant; rSjTPI or the rSjTPI-rSjLD1 combination resulted in a lower level of protection. Mirroring our previous results, there were significant reductions in the number of female worms (30⁻44%), faecal eggs (61⁻68%), liver eggs (44⁻56%), intestinal eggs (46⁻48%) and mature intestinal eggs (58⁻63%) in the rSjLD1-vaccinated mice compared with the adjuvant only groups. At 6-weeks post-cercarial challenge, a significantly increased production of interferon gamma (IFNγ) in rSjLD1-stimulated splenic CD4⁺ T cells was observed in the rSjLD1-vaccinated mice suggesting a Th1-type response is associated with the generated level of protective efficacy.


Assuntos
Imunidade , Receptor de Insulina/imunologia , Proteínas Recombinantes/imunologia , Schistosoma japonicum/imunologia , Esquistossomose Japônica/imunologia , Esquistossomose Japônica/prevenção & controle , Animais , Anticorpos Antiprotozoários/imunologia , Reações Cruzadas/imunologia , Citocinas/metabolismo , Modelos Animais de Doenças , Feminino , Imunofluorescência , Glucose/metabolismo , Imunização , Imunoglobulina G/imunologia , Masculino , Camundongos , Vacinas Protozoárias/imunologia , Esquistossomose Japônica/metabolismo , Subpopulações de Linfócitos T/imunologia , Subpopulações de Linfócitos T/metabolismo
13.
Int J Mol Sci ; 19(8)2018 Aug 16.
Artigo em Inglês | MEDLINE | ID: mdl-30115897

RESUMO

To further investigate the importance of Schistosoma japonicum acetylcholinesterase (SjAChE) in cholinergic signaling for parasite growth and development, we used RNA interference (RNAi) to knock-down its expression in adults and eggs in vitro. This resulted in its reduced transcription but also expression of other important genes involved both in cholinergic signaling and glucose uptake were impacted substantially. Significant decreases in AChE protein expression, AChE enzymatic activity, and glucose uptake were observed in the SjAChE-knockdown parasites compared with luciferase controls. In vaccine/challenge experiments, we found that immunization of mice with recombinant SjAChE (rSjAChE) expressed in Escherichia coli elicited reductions in male worm numbers (33%), liver granuloma density (41%), and reduced numbers of mature intestinal eggs (73%) in the vaccinated group compared with the control group. These results indicate AChE plays an important role in the metabolism of male worms, and impacts indirectly on female fecundity leading to increased numbers of immature eggs being released and reduced sizes of liver granulomas. Furthermore, cytokine analysis showed that immunization of mice with rSjAChE elicited a predominantly Th1-type immune response characterized by increased production of IFNγ in splenic CD4⁺ T cells of vaccinated mice. The study confirms the potential of SjAChE as a vaccine/drug candidate against zoonotic schistosomiasis japonica.


Assuntos
Acetilcolinesterase/metabolismo , Parasitos/enzimologia , Parasitos/crescimento & desenvolvimento , Schistosoma japonicum/enzimologia , Schistosoma japonicum/crescimento & desenvolvimento , Animais , Formação de Anticorpos , Citocinas/metabolismo , Regulação da Expressão Gênica , Técnicas de Silenciamento de Genes , Glucose/metabolismo , Fígado/parasitologia , Fígado/patologia , Camundongos Endogâmicos CBA , Óvulo/metabolismo , Parasitos/genética , Interferência de RNA , RNA de Cadeia Dupla/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Receptores Nicotínicos/genética , Receptores Nicotínicos/metabolismo , Schistosoma japonicum/genética , Esquistossomose Japônica/imunologia , Esquistossomose Japônica/parasitologia , Baço/metabolismo , Transcrição Gênica , Resultado do Tratamento , Vacinação , Vacinas/imunologia
14.
Sheng Li Xue Bao ; 70(2): 123-131, 2018 Apr 25.
Artigo em Zh | MEDLINE | ID: mdl-29691576

RESUMO

Cholinergic receptor activation and intracellular current injection lead to the persistent activity (PA), which may be involved in inducing neural plasticity. Our previous study showed that PA is closely related to the activation of muscarinic acetylcholine receptors (mAChRs) in pyramidal neurons of mouse primary auditory cortex (AI). However, the subtypes of mAChRs involved in PA remain unclear. Thus, using whole-cell patch-clamp recording and pharmacological methods, we investigated the role of different mAChR subtypes in inducing PA in AI layer V pyramidal neurons of young mice. The results showed that activation of mAChRs with intracellular depolarizing current induced PA in layer V pyramidal neurons. Blockade of M1, M2 or M3 subtypes prevented the PA, whereas M4 receptor antagonists did not affect the production of PA. The results suggest that the PA may be induced through a mechanism involving M1, M2 and M3 muscarinic receptors, but not M4 subtype.


Assuntos
Córtex Auditivo/fisiologia , Células Piramidais/fisiologia , Receptores Muscarínicos/fisiologia , Acetilcolina , Animais , Camundongos , Técnicas de Patch-Clamp , Receptores Muscarínicos/classificação
15.
Molecules ; 22(9)2017 Sep 14.
Artigo em Inglês | MEDLINE | ID: mdl-28906438

RESUMO

Schistosomiasis, which is caused by helminth trematode blood flukes of the genus Schistosoma, is a serious health and economic problem in tropical areas, and the second most prevalent parasitic disease after malaria. Currently, there is no effective vaccine available and treatment is entirely dependent on a single drug, praziquantel (PZQ), raising a significant potential public health threat due to the emergence of PZQ drug resistance. It is thus urgent and necessary to explore novel therapeutic targets for the treatment of schistosomiasis. Previous studies demonstrated that acetylcholinesterase (AChE) and nicotinic acetylcholine receptors (nAChRs) play important roles in the schistosome nervous system and ion channels, both of which are targeted by a number of currently approved and marketed anthelminthic drugs. To improve understanding of the functions of the cholinergic system in schistosomes, this article reviews previous studies on AChE and nAChRs in schistosomes and other helminths and discusses their potential as suitable targets for vaccine development and drug design against schistosomiasis.


Assuntos
Acetilcolinesterase/metabolismo , Helmintos/metabolismo , Receptores Nicotínicos/metabolismo , Animais , Anti-Helmínticos/uso terapêutico , Desenho de Fármacos , Resistência a Medicamentos , Humanos , Praziquantel/farmacologia , Schistosoma/metabolismo , Esquistossomose/tratamento farmacológico , Esquistossomose/prevenção & controle , Vacinas
16.
J Proteome Res ; 14(11): 4623-34, 2015 Nov 06.
Artigo em Inglês | MEDLINE | ID: mdl-26370134

RESUMO

Schistosomiasis is a parasitic zoonosis posing great threat to human health. The infection is acquired by larval cercariae penetrating host skin and transforming into juveniles, schistosomula. Proteolytic enzymes secreted from the cercarial acetabular glands are known to aid to the skin penetration, but molecular mechanisms remain largely unclear. To profile the protein composition and identify potential invasive proteases, we developed a new method for simulating cercarial transformation and collecting schistosomula, and for the first time, we compared the proteomes of Schistosoma japonicum cercariae and schistosomula by using in-gel shotgun proteomic analysis. Totally, 1972 proteins were identified in association with ten main biological processes based on Gene Ontology analysis; 46 proteases were detected in cercariae, and among them, 25 proteases disappeared after penetrated. Notably, leishmanolysins and serine and cysteine proteases were found abundant but differentially expressed. Recombinant serine protease SjCE2b and cysteine protease SjCB2 were produced and used for validation of native proteins. Immunofluorescence and Western blotting assays detected SjCE2b and SjCB2 in cercariae but not in schistosomula, suggesting the two enzymes might be consumed upon skin migration. Our data comprehensively chart the proteomic changes during cercarial invasion, revealing the potential proteases involved, providing a platform for the development of molecular anti-infection strategy.


Assuntos
Cercárias/metabolismo , Cisteína Proteases/química , Proteínas de Helminto/química , Fragmentos de Peptídeos/isolamento & purificação , Proteoma/química , Schistosoma japonicum/metabolismo , Serina Endopeptidases/química , Sequência de Aminoácidos , Animais , Cercárias/genética , Cercárias/crescimento & desenvolvimento , Cromatografia Líquida , Cisteína Proteases/genética , Cisteína Proteases/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Feminino , Expressão Gênica , Proteínas de Helminto/genética , Proteínas de Helminto/metabolismo , Estágios do Ciclo de Vida/genética , Camundongos , Camundongos Endogâmicos C57BL , Anotação de Sequência Molecular , Dados de Sequência Molecular , Proteólise , Proteoma/genética , Proteoma/metabolismo , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Schistosoma japonicum/genética , Schistosoma japonicum/crescimento & desenvolvimento , Serina Endopeptidases/genética , Serina Endopeptidases/metabolismo , Pele/parasitologia , Caramujos/parasitologia , Espectrometria de Massas em Tandem
17.
Sheng Li Xue Bao ; 67(4): 370-8, 2015 Aug 25.
Artigo em Zh | MEDLINE | ID: mdl-26300248

RESUMO

The inferior colliculus (IC) is a pivot along the central auditory pathway. Using infrared visual whole-cell patch clamp recording technique, we investigated the electrophysiological properties of IC subnuclei neurons. Recordings were made from 88 neurons, including 21 neurons from the dorsal cortex of the IC (ICd), 43 neurons from the central nucleus of the IC (ICc) and 24 neurons from the external cortex of the IC (ICx). Based on the responses to positive current injection, three firing patterns, i.e., onset (6.8%, n = 6), adapting (39.8%, n = 35) and sustained (53.4%, n = 47) patterns, were identified. The hyperpolarization-activated inward current (Ih) could be recorded in half of the neurons (49/88). The sustained pattern occurred in more than half of ICd and ICc neurons (61.9% and 67.4%), while the adapting pattern occurred in majority of ICx neurons (75%). Action potential (AP) threshold and time constant also showed significant differences across neurons from the ICd, the ICc and the ICx. Our results indicate that IC neurons are different in electrophysiological properties across the subnuclei. The variance of the responses may be related to the distinct types of neurons as well as the received projections, which is implicated in the distinct roles of IC neurons in central auditory processing.


Assuntos
Colículos Inferiores/citologia , Neurônios/citologia , Técnicas de Patch-Clamp , Potenciais de Ação , Animais , Fenômenos Eletrofisiológicos , Técnicas In Vitro , Camundongos
18.
Artigo em Zh | MEDLINE | ID: mdl-25065206

RESUMO

OBJECTIVE: To clone and express the recombinant proteins based on the whole open reading frame of two tyrosinases (tyrosinase 1 and tyrosinase 2) from Schistosoma japonicum, and study the transcription specificity of the two tyrosinases in different sex and developmental stages of S. japonicum. METHODS: The full-length of SjTYR1 and SjTYR2 were amplified with specific primers and subcloned into pSJ2. The recombinant plasmids were transformed into E. coli Rosetta Gami strains and induced with IPTG for expression. The recombinant proteins were purified by Ni-NTA agarose. The recombinant proteins SjTYR1 and SjTYR2 were used to produce the specific antibodies by immunizing the rabbits. The immunogenicity of the recombinant proteins SjTYR1 and SjTYR2 were detected by Western blotting using sera of recombinant proteins-immunized rabbits and S. japonicum-infected rabbit serum as the primary antibody, respectively. The reactivity of sera from recombinant proteins-immunized rabbits was analyzed by Western blotting against the native protein of S. japonicum worm. Total RNA was extracted from 14, 16, 18, 20, 22, 24, 26, and 28-day male and female worms. Transcription levels of the two tyrosinases in different sex and different stage were determined via RT-PCR method. RESULTS: The expression vector of SjTYR1/pSJ2 and SjTYR2/pSJ2 were constructed and the recombinant pro teins SjTYR1 and SjTYR2 were expressed in inclusion body in E. coli (about M(r) 55 000 and M(r) 56 800). The sera of S. japonicum-infected rabbits reacted positively with the purified recombinant protein SjTYR1, but not with recombinant protein SjTYR2. The native protein of S. japonicum worm could be recognized by sera of rSjTYR1-immunized rabbits (M(r) 100 000), but not by sera of rSjTYR2-immunized rabbits. Transcription levels of the two tyrosinases in male worms were nearly zero. In female worms, the transcription levels of the two tyrosinases increased sharply from the 24th day post-in- fection and reached maximum on the 28th day. CONCLUSION: The recombinant proteins of SjTYR1 and SjTYR2 show immunogenicity and immunoreactivity. SjTYR1 and SjTYR2 are both expressed specifically in female worms and the transcription levels increase in 24-28 days after infection.


Assuntos
Monofenol Mono-Oxigenase/metabolismo , Schistosoma japonicum/metabolismo , Transcrição Gênica , Animais , Western Blotting , Clonagem Molecular , Escherichia coli , Feminino , Expressão Gênica , Masculino , Monofenol Mono-Oxigenase/genética , Coelhos , Proteínas Recombinantes , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Schistosoma japonicum/genética
19.
Artigo em Zh | MEDLINE | ID: mdl-25065204

RESUMO

OBJECTIVE: To clone and express recombinant calpain of Schistosoma japonicum (Sjcalpain), observe the distribution of Sjcalpain in S. japonicum cercariae and analyze its role in skin invasion. METHODS: The primers were designed according to the full-length sequence of calpain (GenBank accession No. AB016726). The genes encoding catalytic domain and Ca2+ binding domain of Sjcalpain were amplified by PCR, and the target fragments were subcloned into pET-28a. The recombinant proteins were expressed in E. coli BL21 (DE3) and purified by Ni-NTA resin. The rabbit polyclonal antibodies were prepared with the two purified recombinant proteins by immunizing New Zealand white rabbits. ELISA was used to detect the titer of rabbit antiserum. Immunolocalization was used to investigate the distribution of Sjcalpain in S. japonicum cercariae. Cercariae were incubated with specific inhibitor before infection of mice and the worm reduction rate was calculated. RESULTS: The recombinant expression vector Sjcalpain catalytic domain/pET28a and Sjcalpain Ca2+ binding domain/pET28a were constructed and the recombinant proteins were successfully expressed in E. coli BL21 (DE3) (about M(r) 43 000 and M(r) 39 000, respectively). The two target proteins were expressed as inclusion bodies. The purified target proteins were obtained through Ni-NTA affinity purification. ELISA result showed that the titer of prepared rabbit polyclonal antibodies was higher than 1 : 80 000. Immunolocalization study demonstrated that Sjcalpain protein was mainly expressed in the head of cercariae. Inhibition assays suggested that the average number of adult worms in calpain inhibitor-incubation group and control group was 19 and 23, respectively, with a worm reduction rate of 17.4%. CONCLUSION: Sjcalpain is mainly expressed in the head of S. japonicum cercariae. Inhibition of Sjcalpain could reduce the number of invading cercariae in infected mice, which suggest that Sjcalpain may play a role in skin invasion by cercariae.


Assuntos
Calpaína/metabolismo , Schistosoma japonicum/metabolismo , Esquistossomose Japônica/metabolismo , Animais , Cercárias , Ensaio de Imunoadsorção Enzimática , Escherichia coli , Camundongos , Coelhos , Proteínas Recombinantes , Schistosoma japonicum/genética
20.
Anatol J Cardiol ; 2024 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-38430113

RESUMO

OBJECTIVE: Although telemedicine interventional therapy is an innovative method to reduce public medical burden and improve heart failure, its effectiveness is still controversial. This meta-analysis evaluates the role of telemedicine interventional therapy in the treatment of patients with chronic heart failure. METHODS: Relevant literature on telemedicine in chronic heart failure treatment was screened and extracted based on predefined criteria. Quality assessment used Cochrane Handbook 5.1.0 tool, and meta-analysis was conducted using R 4.2.2 software. RESULTS: Fifteen English-language articles were ultimately included in this meta-analysis. The risk bias evaluation determined that 4 articles were low-risk bias and 11 articles were unclear risk bias. The meta-analysis revealed that, compared to the routine intervention group, the all-cause hospitalization rate of patients in the telemedicine intervention group decreased [OR = 0.63, 95% CI (0.41; 0.96), P =.03], and the hospitalization rate of heart failure also decreased [OR = 0.70, 95% CI (0.48; 0.85), P <.01]. However, there were no differences in mortality [OR = 0.64, 95% CI (0.41; 1.01), P =.05], length of hospitalization [MD = -0.42, 95% CI (-1.22; 0.38), P =.31], number of emergency hospitalizations [MD = -0.09, 95% CI (-0.33; 0.15), P =.45], medication compliance [OR = 1.67, 95% CI (0.92; 3.02), P =.09], or MLHFQ scores [MD = -2.30, 95% CI (-6.16; 1.56), P =.24] among the patients. CONCLUSION: This meta-analysis showed that telemedicine reduced overall and heart failure-related hospitalizations in chronic heart failure patients, suggesting its value in clinical management. However, it did not significantly affect mortality, hospital stay length, emergency visits, medication adherence, or quality of life. This suggests the need to optimize specific aspects of telemedicine, identify key components, and develop strategies for better treatment outcomes.

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