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1.
Biochem Biophys Res Commun ; 712-713: 149939, 2024 Jun 18.
Artigo em Inglês | MEDLINE | ID: mdl-38640729

RESUMO

Human heavy-chain ferritin is a naturally occurring protein with high stability and multifunctionality in biological systems. This study aims to utilize a prokaryotic expression system to produce recombinant human heavy-chain ferritin nanoparticles and investigate their targeting ability in brain tissue. The human heavy-chain ferritin gene was cloned into the prokaryotic expression vector pET28a and transformed into Escherichia coli BL21 (DE3) competent cells to explore optimal expression conditions. The recombinant protein was then purified to evaluate its immunoreactivity and characteristics. Additionally, the distribution of the administered protein in normal mice and its permeability in an in vitro blood-brain barrier (BBB) model were measured. The results demonstrate that the purified protein can self-assemble extracellularly into nano-cage structures of approximately 10 nm and is recognized by corresponding antibodies. The protein effectively penetrates the blood-brain barrier and exhibits slow clearance in mouse brain tissue, showing excellent permeability in the in vitro BBB model. This study highlights the stable expression of recombinant human heavy-chain ferritin using the Escherichia coli prokaryotic expression system, characterized by favorable nano-cage structures and biological activity. Its exceptional brain tissue targeting and slow metabolism lay an experimental foundation for its application in neuropharmaceutical delivery and vaccine development fields.


Assuntos
Barreira Hematoencefálica , Encéfalo , Escherichia coli , Ferritinas , Nanopartículas , Proteínas Recombinantes , Animais , Humanos , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Camundongos , Barreira Hematoencefálica/metabolismo , Encéfalo/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Nanopartículas/química , Ferritinas/metabolismo , Ferritinas/genética , Ferritinas/química , Apoferritinas/metabolismo , Apoferritinas/genética , Apoferritinas/química , Distribuição Tecidual
2.
J Sci Food Agric ; 104(6): 3246-3255, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38081762

RESUMO

BACKGROUND: The aim of this research was to evaluate the possibility of lipid concomitant γ-oryzanol reducing oil absorbency of fried foods and the underlying mechanism. Therefore, the influence of γ-oryzanol on moisture and oil content, and distribution and micromorphology of French fries and the viscosity, fatty acid composition and total polar compounds content of rice bran oil (RBO) after frying were studied. RESULTS: Our results showed that the incorporation of low concentration of γ-oryzanol [low addition group (LAG)] (5.754 g/kg) decreased the oil absorbency and porous structure of French fries during frying. Additionally, LAG incorporation inhibited the degradation of linoleic acid, decreased the growth rate of saturated fatty acids, total polar compounds and viscosity of frying oil. CONCLUSIONS: Consequently, it was recommended to incorporate a small amount of γ-oryzanol in frying oil because it could inhibit oil absorption behavior of French fries. © 2023 Society of Chemical Industry.


Assuntos
Culinária , Fenilpropionatos , Culinária/métodos , Ácidos Graxos , Óleo de Farelo de Arroz
3.
Urol Int ; 107(2): 193-201, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-35671712

RESUMO

INTRODUCTION: Postoperative hypertension resolution among patients with adrenal incidentalomas and normal hormone levels was unknown. Identifying the predictive factors was beneficial to the management of adrenal incidentalomas. METHODS: We conducted a retrospective cohort study, recruiting patients undergoing laparoscopic adrenal tumor resection for adrenal incidentaloma with hypertension and normal hormone levels. Demographic, clinical, treatment, and laboratory data were collected and compared. We used univariable and multivariable logistic regression methods to identify the predictive factors of postoperative hypertension resolution. RESULTS: Of the 171 patients in our study, 130 (76.0%) patients performed a resolution of hypertension, and 57 (33.3%) patients had a significant reduction. Multivariate logistic regression analysis showed that the male sex (odds ratio (OR) 0.305, 95% confidence interval (CI): 0.098-0.948, p = 0.040), body mass index (BMI) (OR 0.973, 95% CI: 0.670-0.938, p = 0.007), aldosterone and plasma renin activity ratio (APR) in erect position (OR 1.206, 95% CI: 1.042-1.397, p = 0.012), and preoperative systolic pressure (OR 1.044, 95% CI: 1.009-1.080, p = 0.014), were significantly associated with the outcomes of hypertension resolution. DISCUSSION/CONCLUSION: Adrenal incidentalomas patients with hypertension and normal hormone levels would perform hypertension resolution after laparoscopic adrenal tumor resection, especially for females with low BMI, high preoperative systolic blood pressure, and high APR (erect position).


Assuntos
Neoplasias das Glândulas Suprarrenais , Hipertensão , Laparoscopia , Feminino , Humanos , Masculino , Neoplasias das Glândulas Suprarrenais/complicações , Neoplasias das Glândulas Suprarrenais/cirurgia , Estudos Retrospectivos , Hipertensão/complicações , Aldosterona
4.
Eur Arch Otorhinolaryngol ; 280(4): 1919-1926, 2023 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-36422670

RESUMO

OBJECTIVE: To determine the value of a questionnaire as a screening tool for benign paroxysmal position vertigo (BPPV). STUDY DESIGN: Retrospective chart review. SETTING: Tertiary care centers. METHODS: A total of 520 vertigo adults completed the questionnaire before the diagnosis was confirmed. After vestibular function examination and other diagnostic examination, the diagnosis of all participants was confirmed by experts. By validating valuable items from the questionnaire with 47 items, a new questionnaire of 5 items was formed to quickly diagnose BPPV. The internal consistency of the new questionnaire and validity were evaluated. The correlation between the score obtained from the new questionnaire and diagnosis was investigated. The mean score was also compared between groups with and without BPPV and diagnostic precision measures were calculated. RESULTS: 520 vertigo participants answered all the question completely and BPPV was identified in 138 participants (26.5%). The responses to questionnaire revealed preferable reproducibility (r = 0.898, P < 0.05) and internal consistency (Cronbach's α = 0.702) as well as the validity (Kaiser-Meyer-Olkin, KMO = 0.731). The higher the individual score, the more likely to be BPPV (B = 2.082; P < 0.05). The mean score of answers was greater in the group with a clinical diagnosis of BPPV compared to those without BPPV (F = 58.459, P < 0.05). The sensitivity of the screening tool was 92.8% and specificity was 88.5%, with an area under the ROC curve of 0.946 (95% confidence interval 0.926-0.965; P < 0.05). CONCLUSION: The questionnaire proved to be of great value to screen for individuals with possible BPPV.


Assuntos
Vertigem Posicional Paroxística Benigna , Adulto , Humanos , Estudos Retrospectivos , Reprodutibilidade dos Testes , Vertigem Posicional Paroxística Benigna/diagnóstico , Inquéritos e Questionários , Curva ROC
5.
Molecules ; 28(5)2023 Feb 26.
Artigo em Inglês | MEDLINE | ID: mdl-36903419

RESUMO

The acidic extracellular microenvironment has become an effective target for diagnosing and treating tumors. A pH (low) insertion peptide (pHLIP) is a kind of peptide that can spontaneously fold into a transmembrane helix in an acidic microenvironment, and then insert into and cross the cell membrane for material transfer. The characteristics of the acidic tumor microenvironment provide a new method for pH-targeted molecular imaging and tumor-targeted therapy. As research has increased, the role of pHLIP as an imaging agent carrier in the field of tumor theranostics has become increasingly prominent. In this paper, we describe the current applications of pHLIP-anchored imaging agents for tumor diagnosis and treatment in terms of different molecular imaging methods, including magnetic resonance T1 imaging, magnetic resonance T2 imaging, SPECT/PET, fluorescence imaging, and photoacoustic imaging. Additionally, we discuss relevant challenges and future development prospects.


Assuntos
Neoplasias , Medicina de Precisão , Humanos , Peptídeos/química , Imageamento por Ressonância Magnética , Concentração de Íons de Hidrogênio , Microambiente Tumoral
6.
BMC Urol ; 22(1): 128, 2022 Aug 20.
Artigo em Inglês | MEDLINE | ID: mdl-35987640

RESUMO

BACKGROUND: Combining immune checkpoint inhibitors with chemotherapy can synergistically improve antitumor activity and are generally well tolerated. Recently, the efficacy and safety of combination therapy has been demonstrated for many cancers, including urothelial carcinomas. The aim of this retrospective pilot study was to evaluate the efficacy and safety of tislelizumab plus chemotherapy as first-line adjuvant treatment for locally advanced or metastatic bladder cancer. METHODS: We conducted a retrospective analysis of 31 patients with locally advanced or metastatic bladder cancer from December 2020 to January 2022 with an Eastern Cooperative Oncology Group performance status of 0/1. Of the 31 patients, 14 patients received tislelizumab (200 mg i.v. every 3 weeks, Q3W) plus 21 days cycles of chemotherapy (gemcitabine, 1000 mg/m2 i.v. on days 1 and 8 of each cycle + cisplatin, 70 mg/m2 i.v. on day 2 of each cycle) (TGC) treatment and 17 patients received gemcitabine plus cisplatin chemotherapy (GC) treatment. All patients treated with bladder cytoreductive surgery and were treated for four 21 days cycles until disease progression or intolerable treatment-related adverse events (TRAEs). The objective progression-free survival (PFS), overall survival (OS), overall response rate (ORR), disease control rate (DCR), clinical benefit rate (CBR) and TRAEs were recorded and reviewed. RESULTS: As of the cut-off date (March 25, 2022), PFS, OS, ORR, DCR, CBR and TRAEs were evaluated in 14 patients receiving combination therapy and 17 patients in the chemotherapy alone group. The median PFS was 36.0 [95% confidence interval (CI) 33.1-38.9] weeks in the TGC group and 29.0 (95% CI 25.4-32.6) weeks in the GC group [hazard ratio (HR) 0.15 (95% CI 0.04-0.55)]. In the GC group, the median OS was 48.0 (95% CI 39.7-56.3) weeks; the median OS was not yet mature for the TGC group [HR 0.26 (95% CI 0.07-0.94)]. Treatment with TGC resulted in improved DCR (TGC 71.4%; GC 65.0%) and CBR (TGC 64.3%; GC 52.9%) compared with GC. However, although higher incidences of grade ≥ 3 TRAEs were observed with TGC compared with GC (35.7% vs 23.5%), the difference was not statistically significant (p = 0.47). CONCLUSION: This study suggested that TGC provided survivors of locally advanced or metastatic bladder cancer with encouraging antitumor activity and was generally well tolerated.


Assuntos
Cisplatino , Neoplasias da Bexiga Urinária , Anticorpos Monoclonais Humanizados , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Desoxicitidina/análogos & derivados , Humanos , Projetos Piloto , Estudos Retrospectivos , Resultado do Tratamento , Neoplasias da Bexiga Urinária/tratamento farmacológico , Neoplasias da Bexiga Urinária/patologia , Gencitabina
7.
Molecules ; 27(23)2022 Nov 29.
Artigo em Inglês | MEDLINE | ID: mdl-36500432

RESUMO

Aminopeptidase N (APN) was closely associated with cancer invasion, metastasis, and angiogenesis. Therefore, APN inhibitors have attracted more and more attention of scientists as antitumor agents. In the current study, we designed, synthesized, and evaluated one new series of pyrazoline-based hydroxamate derivatives as APN inhibitors. Moreover, the structure-activity relationships of those were discussed in detail. 2,6-Dichloro substituted compound 14o with R1 = CH3, showed the best capacity for inhibiting APN with an IC50 value of 0.0062 ± 0.0004 µM, which was three orders of magnitude better than that of the positive control bestatin. Compound 14o possessed both potent anti-proliferative activities against tumor cells and potent anti-angiogenic activity. At the same concentration of 50 µM, compound 14o exhibited much better capacity for inhibiting the micro-vessel growth relative to bestatin in the rat thoracic aorta ring model. Additionally, the putative interactions of 14o with the active site of APN are also discussed. The hydroxamate moiety chelated the zinc ion and formed four hydrogen bonds with His297, Glu298 and His301. Meanwhile, the terminal phenyl group and another phenyl group of 14o interacted with S2' and S1 pockets via hydrophobic effects, respectively.


Assuntos
Antineoplásicos , Antígenos CD13 , Ratos , Animais , Inibidores de Proteases/química , Ácidos Hidroxâmicos/farmacologia , Antineoplásicos/farmacologia , Relação Estrutura-Atividade
8.
Pharmacol Res ; 165: 105442, 2021 03.
Artigo em Inglês | MEDLINE | ID: mdl-33497805

RESUMO

Epidermal growth factor receptor (EGFR) T790M mutation act as the dominant resistance mechanism to first and second generations tyrosine kinase inhibitors (TKIs), the roles of miR-7 in the development of T790M mutation are largely unknown. Here, we confirmed that the level of miR-7 was significantly higher in the gefitinib sensitivity PC9 cells compared to gefitinib resistance H1975 cells, and miR-7 overexpression promoted the apoptosis of H1975 cells by gefitinib treatment. Furthermore, we found that exosomes could transfer miR-7 mimics from PC9 cells to H1975 cells, which reversed gefitinib resistance through binding to YAP, and altered H1975 cells resistance phenotype in vitro. In addition, we suppressed exosomal miR-7 by GW4869, increasing PC9 cells chemoresistance to gefitinib treatment in vivo. Of note, we detected that miR-7 was significantly higher in serum exosomes from healthy controls than from patients with lung carcinoma, and high miR-7 expression was associated with strong response to lung carcinoma patients receiving gefitinib treatment, as well as a longer survival. Therefore, exosomal miR-7 can act as a potential biomarker and therapeutic target for EGFR T79M resistance mutations.


Assuntos
Carcinoma Pulmonar de Células não Pequenas/sangue , Proteínas de Ciclo Celular/metabolismo , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Exossomos/metabolismo , Gefitinibe/uso terapêutico , Neoplasias Pulmonares/sangue , MicroRNAs/sangue , Fatores de Transcrição/metabolismo , Idoso , Animais , Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Biomarcadores Tumorais/biossíntese , Biomarcadores Tumorais/sangue , Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Linhagem Celular Tumoral , Resistencia a Medicamentos Antineoplásicos/fisiologia , Exossomos/efeitos dos fármacos , Feminino , Gefitinibe/farmacologia , Humanos , Neoplasias Pulmonares/tratamento farmacológico , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , MicroRNAs/biossíntese , Pessoa de Meia-Idade , Inibidores de Proteínas Quinases/farmacologia , Inibidores de Proteínas Quinases/uso terapêutico , Carga Tumoral/efeitos dos fármacos , Carga Tumoral/fisiologia , Ensaios Antitumorais Modelo de Xenoenxerto
9.
Nanomedicine ; 32: 102342, 2021 02.
Artigo em Inglês | MEDLINE | ID: mdl-33253922

RESUMO

Acute kidney injury (AKI) is a life-threatening disease without effective treatment. The utilization of curcumin (Cur) for the treatment of AKI is still facing challenges due to its poor water-solubility and low bioavailability. Herein, kidney-targeted octenyl succinic anhydride-grafted fucoidan loaded with Cur (OSA-Fucoidan/Cur) was fabricated for synergistic treatment of AKI. It was found that OSA-Fucoidan/Cur micelles had a sustained drug release behavior and excellent physicochemical stability. Cellular uptake studies demonstrated that the specific binding between fucoidan and P-selectin overexpressed on H2O2-stimulated HUVECs contributed to the higher internalization of OSA-Fucoidan/Cur micelles by the cells. In addition, OSA-Fucoidan micelles exhibited an ideal kidney-targeted characteristic in lipopolysaccharide (LPS)-induced AKI mice. In vivo studies showed that the combination of Cur and OSA-Fucoidan endowed the OSA-Fucoidan/Cur micelles with synergistically anti-inflammatory and antioxidant abilities, thereby largely enhancing the therapeutic efficacy of AKI. Therefore, OSA-Fucoidan/Cur micelles may represent a potential kidney-targeted nanomedicine for effective treatment of AKI.


Assuntos
Injúria Renal Aguda/tratamento farmacológico , Portadores de Fármacos/química , Micelas , Selectina-P/antagonistas & inibidores , Polissacarídeos/química , Injúria Renal Aguda/patologia , Animais , Antioxidantes/farmacologia , Curcumina/farmacologia , Curcumina/uso terapêutico , Liberação Controlada de Fármacos , Endocitose/efeitos dos fármacos , Meia-Vida , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , Masculino , Camundongos Endogâmicos ICR , Anidridos Succínicos/química , Distribuição Tecidual/efeitos dos fármacos
10.
J Org Chem ; 85(17): 11265-11279, 2020 09 04.
Artigo em Inglês | MEDLINE | ID: mdl-32701277

RESUMO

We report, herein, the synthesis of thieno[3,2-b]pyrroles from the direct oxidative [4 + 1] cyclization of 2-alkynyl pyrrolidines with elemental sulfur. This transformation likely originates from electrophilic attack at the ß-position of pyrrolidine followed by an intramolecular thienannulation to deliver the desired product. Mechanistic investigation suggests that the present reaction involves the formation of dihydrothieno[3,2-b]pyrrole as an intermediate.

11.
Eur J Clin Microbiol Infect Dis ; 39(12): 2309-2315, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-32683596

RESUMO

During the COVID-19 outbreak, the mobile cabin hospital has effectively isolated and treated patients diagnosed as mild-moderate disease. However, a detailed clinical course has not been well described. We included 483 patients who were isolated and treated from Feb 6, 2020, to Feb 15, 2020, including definite outcome (discharge or deterioration). Sixty-two patients were transferred to severe cases, of whom were trasfered to designated hospital for intensive care. By March 9, 2020, all patients were discharged without dead. The mobile cabin hospital provides feasible strategy of isolation of mild-moderate cases and timely intervention during the virus outbreak.


Assuntos
Doença das Coronárias/diagnóstico , Infecções por Coronavirus/diagnóstico , Pneumopatias/diagnóstico , Pandemias , Alta do Paciente/estatística & dados numéricos , Isolamento de Pacientes/métodos , Pneumonia Viral/diagnóstico , Idoso , Betacoronavirus/patogenicidade , Índice de Massa Corporal , COVID-19 , Teste para COVID-19 , China/epidemiologia , Técnicas de Laboratório Clínico , Comorbidade , Doença das Coronárias/epidemiologia , Doença das Coronárias/fisiopatologia , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/fisiopatologia , Feminino , Hospitalização/estatística & dados numéricos , Humanos , Pneumopatias/epidemiologia , Pneumopatias/fisiopatologia , Masculino , Pessoa de Meia-Idade , Unidades Móveis de Saúde , Pneumonia Viral/epidemiologia , Pneumonia Viral/fisiopatologia , Estudos Retrospectivos , SARS-CoV-2 , Índice de Gravidade de Doença
12.
Cell Mol Life Sci ; 76(23): 4613-4633, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31352532

RESUMO

Lung cancer remains the leading cause of cancer-related death worldwide, and the high incidence rates are worrisome. Exosomes are a class of extracellular vesicles secreted by most cells, including RNAs, proteins and lipids. Exosomes can mediate cell-to-cell communication in both physiologic and pathologic processes. Accumulated evidences show that cancer-derived exosomes aid in the recruitment and reprogramming of constituents correlated with tumor microenvironment. Furthermore, exosome-based clinical trials have been completed in advanced lung cancer patients. In this review, we discuss the roles of exosomes in a lung cancer microenvironment, such as its participation in lung cancer initiation, progression and metastasis as well as being involved in angiogenesis, epithelial-mesenchymal transition (EMT), immune escape, and drug resistance. In addition, we focus on the potential of exosomes as diagnostic and prognostic biomarkers in lung cancer, as well as the challenges faced by and advantages of exosomes as drug delivery vehicles and in exosome-based immunotherapy.


Assuntos
Exossomos/metabolismo , Neoplasias Pulmonares/patologia , Comunicação Celular , Resistencia a Medicamentos Antineoplásicos , Transição Epitelial-Mesenquimal , Humanos , Terapia de Imunossupressão , Neoplasias Pulmonares/irrigação sanguínea , Neoplasias Pulmonares/metabolismo , Metástase Neoplásica , Neovascularização Patológica , Microambiente Tumoral
13.
J Org Chem ; 84(20): 12946-12959, 2019 Oct 18.
Artigo em Inglês | MEDLINE | ID: mdl-31389694

RESUMO

We report a base-promoted metal-free approach for the synthesis of thieno[2,3-b]indole derivatives. This method combined four C-H σ-bond cleavage reactions of two different kinds of C-H bonds and two C-S σ-bond formation processes. A series of thieno[2,3-b]indoles were obtained starting from 3-benzylindole derivatives with good yields and high regioselectivity, with the elemental sulfur serving as a cheap and readily available sulfur source. Good efficiency could be maintained even when the reaction was performed on a gram scale. A plausible mechanism was proposed on the basis of mechanistic studies.

14.
Avian Pathol ; 48(4): 343-351, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30958706

RESUMO

The recombinant Muscovy duck parvovirus (rMDPV) has been recently characterized and identified in China. However, whether other additional rMDPV field isolates exist, and whether these strains possess common molecular characteristics, remain to be explored. In this retrospective study, two new rMDPV isolates, namely, JH06 and JH10, were identified through genome sequencing and recombination analysis. JH06, JH10, and four previously characterized rMDPV strains (SAAS-SHNH, ZW, FJM3, and PT97) underwent the same recombination events in a 1.1-kb region in their VP3 genes and displayed highly consistent beginning and ending breakpoints. JH06, JH10, SAAS-SHNH, ZW, and FJM3, but not PT97, underwent recombination in their P9 promoter regions. In both recombination events, the classical MDPV strain YY acted as the major parent, whereas the virulent strain DY16 and the vaccine strain SYG61v of goose parvovirus (GPV) served as the minor parents. The sequence alignments of inverted terminal repeats (ITRs) revealed that rMDPV strains shared higher identities (96.0%-97.2%) with classical MDPV strains than with GPV and contained typical one-nucleotide-pair deletions in the palindromic stems of their ITRs. This work elucidated the common molecular characteristics and differences of six rMDPV strains. The results of this work will facilitate the preparation of an efficacious vaccine for the protection of Muscovy ducks against rMDPV infection.


Assuntos
Dependovirus/genética , Patos , Infecções por Parvoviridae/veterinária , Doenças das Aves Domésticas/virologia , Sequência de Aminoácidos , Animais , Sequência de Bases , China/epidemiologia , Dependovirus/isolamento & purificação , Dependovirus/patogenicidade , Dados de Sequência Molecular , Infecções por Parvoviridae/epidemiologia , Infecções por Parvoviridae/virologia , Parvovirus/genética , Parvovirus/isolamento & purificação , Parvovirus/patogenicidade , Filogenia , Doenças das Aves Domésticas/epidemiologia , Doenças das Aves Domésticas/prevenção & controle , Distribuição Aleatória , Recombinação Genética , Estudos Retrospectivos , Alinhamento de Sequência/veterinária , Vacinas Virais/normas
15.
Cell Physiol Biochem ; 49(2): 662-677, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30165358

RESUMO

BACKGROUND/AIMS: Phagocytosis of bacteria by monocytes/macrophages can trigger the immune response and the clearance of bacteria. This innate immune response involves Toll-like receptor 4 (TLR4). However, much remains unknown about the mechanism of TLR4-regulated phagocytosis of Salmonella enterica serovar Typhimurium (S. typhimurium) within sheep monocytes/macrophages. Here, we aimed to address these knowledge gaps by infecting transgenic sheep overexpressing TLR4 with S. typhimurium and examining the phagocytic mechanisms involved. METHODS: Transgenic sheep were generated by microinjection of the constructed plasmids into fertilized eggs. Monocytes/macrophages isolated from sheep blood were stimulated with LPS and S. typhimurium. Phagocytosis-related factor expression, phagocytic ability, and adhesion were then determined. TLR4/phosphatidylinositide 3-kinase (PI3K) signaling was inhibited to investigate if this pathway is involved in changes in bacterial internalization in sheep. RESULTS: We found that TLR4 overexpression effectively activated the PI3K signaling pathway and upregulated the expression of scavenger receptors. Additionally, actin polymerization and adhesive capacity were both enhanced in TLR4-overexpressing sheep monocytes/macrophages. TLR4 inhibition decreased S. typhimurium phagocytosis by reducing the actin polymerization and adhesive capacity of cells. Furthermore, inhibition of PI3K markedly impaired TLR4-dependent phagocytosis by restraining actin polymerization and scavenger receptor expression and reduced the adhesive capacity of the monocytes/macrophages. CONCLUSION: Our findings indicate that overexpression of TLR4 enhances phagocytosis through PI3K signaling and the subsequent activation of actin polymerization and scavenger receptors in sheep monocytes/macrophages infected with S. typhimurium.


Assuntos
Macrófagos/imunologia , Fagocitose , Fosfatidilinositol 3-Quinases/metabolismo , Salmonella typhimurium/metabolismo , Receptor 4 Toll-Like/metabolismo , Animais , Animais Geneticamente Modificados/metabolismo , Antígenos CD36/metabolismo , Células Cultivadas , Cromonas/farmacologia , Leucócitos Mononucleares/citologia , Lipopolissacarídeos/imunologia , Lipopolissacarídeos/farmacologia , Macrófagos/citologia , Macrófagos/efeitos dos fármacos , Morfolinas/farmacologia , Fator 88 de Diferenciação Mieloide/metabolismo , Fagocitose/efeitos dos fármacos , Inibidores de Fosfoinositídeo-3 Quinase , Proteínas Proto-Oncogênicas c-akt/metabolismo , Ovinos , Transdução de Sinais/efeitos dos fármacos , Receptor 4 Toll-Like/genética
16.
BMC Vet Res ; 13(1): 327, 2017 Nov 09.
Artigo em Inglês | MEDLINE | ID: mdl-29121936

RESUMO

BACKGROUND: Muscovy duck parvovirus (MDPV) and Goose parvovirus (GPV) are important etiological agents for Muscovy duck parvoviral disease and Derzsy's disease, respectively; both of which can cause substantial economic losses in waterfowl industry. In contrast to GPV, the complete genomic sequence data of MDPV isolates are still limited and their phylogenetic relationships largely remain unknown. In this study, the entire genome of a pathogenic MDPV strain ZW, which was isolated from a deceased Muscovy duckling in 2006 in China, was cloned, sequenced, and compared with that of other classical MDPV and GPV strains. RESULTS: The genome of strain ZW comprises of 5071 nucleotides; this genome was shorter than that of the pathogenic MDPV strain YY (5075 nt). All the four deleted nucleotides produced in strain ZW are located at the base-pairing positions in the palindromic stem of inverted terminal repeats (ITR) without influencing the formation of a hairpin structure. Recombination analysis revealed that strain ZW originated from genetic recombination between the classical MDPV and GPV strain. The YY strain of MDPV acts as the major parent, whereas the virulent strains YZ99-6 and B and the vaccine strain SYG61v of GPV act as the minor parents in varying degrees. Two recombination sites were detected in strain ZW, with the small recombination site surrounding the P9 promoter, and the large recombination site situated in the middle of the VP3 gene. The SYG61V strain is a vaccine strain used for preventing goose parvoviral disease. This strain was found to be solely involved in the recombination event detected in the P9 promoter region. Phylogenetic analyses between strain ZW and other classical strains of MDPV and GPV were performed. The results supported the in silico recombination analysis conclusion. CONCLUSIONS: MDPV Strain ZW is a novel recombinant parvovirus, and the bulk of its genome originates from the classical MDPV strain. Two virulent strains and a vaccine strain of GPV were involved in the recombination process in varying degrees.


Assuntos
Genoma Viral , Parvovirinae/genética , Animais , China , Patos , Infecções por Parvoviridae/veterinária , Infecções por Parvoviridae/virologia , Parvovirinae/classificação , Filogenia , Doenças das Aves Domésticas/virologia , Recombinação Genética , Análise de Sequência de DNA
17.
Int J Biol Sci ; 20(5): 1905-1926, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38481802

RESUMO

Increasing evidence suggests that autophagy plays a major role during renal fibrosis. Transcription factor EB (TFEB) is a critical regulator of autophagy- and lysosome-related gene transcription. However, the pathophysiological roles of TFEB in renal fibrosis and fine-tuned mechanisms by which TFEB regulates fibrosis remain largely unknown. Here, we found that TFEB was downregulated in unilateral ureteral obstruction (UUO)-induced human and mouse fibrotic kidneys, and kidney-specific TFEB overexpression using recombinant AAV serotype 9 (rAAV9)-TFEB in UUO mice alleviated renal fibrosis pathogenesis. Mechanically, we found that TFEB's prevention of extracellular matrix (ECM) deposition depended on autophagic flux integrity and its subsequent blockade of G2/M arrest in tubular cells, rather than the autophagosome synthesis. In addition, we together RNA-seq with CUT&Tag analysis to determine the TFEB targeted gene ATP6V0C, and revealed that TFEB was directly bound to the ATP6V0C promoter only at specific site to promote its expression through CUT&Run-qPCR and luciferase reporter assay. Interestingly, TFEB induced autophagic flux integrity, mainly dependent on scaffold protein ATP6V0C-mediated autophagosome-lysosome fusion by bridging with STX17 and VAMP8 (major SNARE complex) by co-immunoprecipitation analysis, rather than its mediated lysosomal acidification and degradation function. Moreover, we further investigated the underlying mechanism behind the low expression of TEFB in UUO-induced renal fibrosis, and clearly revealed that TFEB suppression in fibrotic kidney was due to DNMT3a-associated TFEB promoter hypermethylation by utilizing methylation specific PCR (MSP) and bisulfite-sequencing PCR (BSP), which could be effectively recovered by 5-Aza-2'-deoxycytidine (5A-za) to alleviate renal fibrosis pathogenesis. These findings reveal for the first time that impaired TFEB-mediated autophagosome-lysosome fusion disorder, tubular cell G2/M arrest and renal fibrosis appear to be sequentially linked in UUO-induced renal fibrosis and suggest that DNMT3a/TFEB/ATP6V0C may serve as potential therapeutic targets to prevent renal fibrosis.


Assuntos
Nefropatias , Obstrução Ureteral , ATPases Vacuolares Próton-Translocadoras , Animais , Humanos , Camundongos , Apoptose , Autofagia/genética , Fatores de Transcrição de Zíper de Leucina e Hélice-Alça-Hélix Básicos/genética , Fatores de Transcrição de Zíper de Leucina e Hélice-Alça-Hélix Básicos/metabolismo , Linhagem Celular Tumoral , Fibrose , Pontos de Checagem da Fase G2 do Ciclo Celular , Nefropatias/metabolismo , Lisossomos/metabolismo , Proteínas SNARE/metabolismo , Proteínas SNARE/farmacologia , Obstrução Ureteral/metabolismo , ATPases Vacuolares Próton-Translocadoras/metabolismo , ATPases Vacuolares Próton-Translocadoras/farmacologia
18.
Virulence ; 15(1): 2366874, 2024 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-38869140

RESUMO

Recombinant Muscovy duck parvovirus (rMDPV) is a product of genetic recombination between classical Muscovy duck parvovirus (MDPV) and goose parvovirus (GPV). The recombination event took place within a 1.1-kb DNA segment located in the middle of the VP3 gene, and a 187-bp sequence extending from the P9 promoter to the 5' initiation region of the Rep1 ORF. This resulted in the alteration of five amino acids within VP3. Despite these genetic changes, the precise influence of recombination and amino acid mutations on the pathogenicity of rMDPV remains ambiguous. In this study, based on the rMDPV strain ZW and the classical MDPV strain YY, three chimeric viruses (rZW-mP9, rZW-mPR187, and rYY-rVP3) and the five amino acid mutations-introduced mutants (rZW-g5aa and rYY-5aa(ZW)) were generated using reverse genetic technology. When compared to the parental virus rZW, rZW-g5aa exhibited a prolonged mean death time (MDT) and a decreased median lethal dose (ELD50) in embryonated duck eggs. In contrast, rYY-5aa(ZW) did not display significant differences in MDT and ELD50 compared to rYY. In 2-day-old Muscovy ducklings, infection with rZW-g5aa and rYY-5aa(ZW) resulted in mortality rates of only 20% and 10%, respectively, while infections with the three chimeric viruses (rZW-mP9, rZW-mPR187, rYY-rVP3) and rZW still led to 100% mortality. Notably, rYY-rVP3, containing the VP3 region from strain ZW, exhibited 50% mortality in 6-day-old Muscovy ducklings and demonstrated significant horizontal transmission. Collectively, our findings indicate that recombination and consequent amino acid changes in VP3 have a synergistic impact on the heightened virulence of rMDPV in Muscovy ducklings.


Assuntos
Proteínas do Capsídeo , Patos , Infecções por Parvoviridae , Mutação Puntual , Doenças das Aves Domésticas , Recombinação Genética , Animais , Virulência , Infecções por Parvoviridae/virologia , Infecções por Parvoviridae/veterinária , Doenças das Aves Domésticas/virologia , Proteínas do Capsídeo/genética , Parvovirinae/genética , Parvovirinae/patogenicidade
19.
Front Neurosci ; 17: 1219556, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37496735

RESUMO

After regular rehabilitation training, paralysis sequelae can be significantly reduced in patients with limb movement disorders caused by stroke. Rehabilitation assessment is the basis for the formulation of rehabilitation training programs and the objective standard for evaluating the effectiveness of training. However, the quantitative rehabilitation assessment is still in the experimental stage and has not been put into clinical practice. In this work, we propose improved spatial-temporal graph convolutional networks based on precise posture measurement for upper limb rehabilitation assessment. Two Azure Kinect are used to enlarge the angle range of the visual field. The rigid body model of the upper limb with multiple degrees of freedom is established. And the inverse kinematics is optimized based on the hybrid particle swarm optimization algorithm. The self-attention mechanism map is calculated to analyze the role of each upper limb joint in rehabilitation assessment, to improve the spatial-temporal graph convolution neural network model. Long short-term memory is built to explore the sequence dependence in spatial-temporal feature vectors. An exercise protocol for detecting the distal reachable workspace and proximal self-care ability of the upper limb is designed, and a virtual environment is built. The experimental results indicate that the proposed posture measurement method can reduce position jumps caused by occlusion, improve measurement accuracy and stability, and increase Signal Noise Ratio. By comparing with other models, our rehabilitation assessment model achieved the lowest mean absolute deviation, root mean square error, and mean absolute percentage error. The proposed method can effectively quantitatively evaluate the upper limb motor function of stroke patients.

20.
Front Med (Lausanne) ; 10: 1185539, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37275385

RESUMO

Purpose: Lower urinary symptoms (LUTS) may persist in a proportion of patients with benign prostatic hyperplasia (BPH) following transurethral resection of prostate (TURP), which is a major cause of reduced quality-of-life. We aimed to investigate the effect of frailty on LUTS in patients with BPH treated with TURP. Methods: We longitudinally evaluated LUTS and health-related quality-of-life (HRQOL) in patients with BPH treated with TURP from February 2019 and January 2022 using International Prostate Symptom Score (IPSS) and Short Form-8 (SF-8), respectively. Patients were divided into frail and non-frail groups according to the Fried phenotype (FP). The primary purpose was comparing the outcomes of LUTS and HRQOL between two groups. Secondary purposes were investigating the frailty as a preoperative predictor of postoperative adverse LUTS outcomes following TURP using logistic regression analysis. A 1:2 propensity score matching (PSM) was performed to reduce the effects of selection bias and potential confounders. Results: Of the 567 patients enrolled, 495 (87.3%) patients were non-frail (FP = 0-2), and the remaining 72 (12.7%) patients were classified into the frail group. There were no significant differences in body mass index (BMI), urine white blood cell (UWBC), creatinine, prostate-specific antigen (PSA) and prostate volume in both groups at baseline (all p > 0.05). However, patients with frailty were older, higher comorbidity rates, lower peak flow rates and lower HRQOL. In the frail group, although LUTS and HRQOL at 6 months following TURP improved significantly compared to those at baseline, it did not show a significant improvement compared with the non-frail group (both p < 0.001). Moreover, multivariable logistic regression analysis demonstrated that preoperative frailty was significantly associated with poor LUTS improvement in both the entire cohort and PSM subset (both p < 0.05), whereas age and comorbidities were not after PSM analysis. Conclusion: In patients with frail or non-frail, TURP for BPH provides overall good results. However, frail individuals are at higher risk of postoperative adverse LUTS outcomes. Frailty has the potential to be a strong objective tool for risk stratification and should be considered during the perioperative evaluation.

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