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1.
Cell Mol Neurobiol ; 2020 May 18.
Artigo em Inglês | MEDLINE | ID: mdl-32424775

RESUMO

Activation of pro-inflammatory microglia is an important mechanism of the cerebral ischemia-reperfusion (I/R)-induced neuronal injury and dysfunction. Mesenchymal stem cells (MSCs) together with their paracrine factors demonstrated curative potential in immune disorders and inflammatory diseases, as well as in ischemic diseases. However, it remains unclear whether conditioned medium from MSCs could effectively regulate the activation and polarization of microglia exposed to I/R stimulation. In this study, we investigated the effects of conditioned medium from bone marrow MSCs (BMSCs-CM) on I/R-stimulated microglia and the potential mechanism involved, as well as the way to obtain more effective BMSCs-CM. First, cell model of oxygen-glucose deprivation/reoxygenation (OGD/R) was established in microglia to mimic the I/R. BMSCs-CM from different culture conditions (normoxic: 21% O2; hypoxic: 1% O2; hypoxia preconditioning: preconditioning with 1% O2 for 24 h) was used to treat the microglia. Our results showed that BMSCs-CM effectively promoted the survival and alleviated the injury of microglia. Moreover, in microglia exposed to OGD/R, BMSCs-CM inhibited significantly the expression of pro-inflammatory cytokines (TNF-α, IL-1ß, and IL-6), CD86 and inducible nitric oxide synthase, whereas upregulated the levels of anti-inflammatory cytokine (IL-10), CD206 and Arginase-1. These results suggested that BMSCs-CM promoted the polarization of anti-inflammatory microglia. In particular, BMSCs-CM from cultures with hypoxia preconditioning was more effective in alleviating cell injury and promoting anti-inflammatory microglia polarization than BMSCs-CM from normoxic cultures and from hypoxic cultures. Furthermore, inhibition of exosomes secretion could largely mitigate these effects of BMSCs-CM. In conclusion, our results suggested that hypoxia preconditioning of BMSCs could enhance the efficacy of BMSCs-CM in alleviating OGD/R-induced injury and in promoting the anti-inflammatory polarization of microglia, and these beneficial effects of BMSCs-CM owed substantially to exosomes.

2.
Int J Med Sci ; 17(8): 1056-1061, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32410835

RESUMO

The aim of this study was to determine whether interleukin-1ß (IL-1ß) promotes oxidised low-density lipoprotein (Ox-LDL) uptake by human glomerular mesangial cells (HMCs) and its effect on the expression of lectin-like Ox-LDL receptor 1 (LOX-1) and to identify pathways through which IL-1ß affects lipid uptake. Confocal laser scanning microscopy and flow cytometry were used to observe the effect of IL-1ß on lipid uptake by HMCs and the pathway by which IL-1ß may mediate lipid uptake. Real-time polymerase chain reaction (PCR) and western blotting were used to evaluate the effect of IL-1ß on LOX-1 expression. Confocal laser scanning microscopy and flow cytometry revealed that IL-1ß promoted uptake of fluorescent Dil-labelled Ox-LDL(Dil-Ox-LDL) by HMCs and the enhanced uptake of Dil-Ox-LDL was partially inhibited by an anti-LOX-1 antibody evaluated by flow cytometry. Further, IL-1ß promoted LOX-1 mRNA and protein expression of HMCs in a dose- and time-dependent manner. Thus, Ox-LDL is ingested by HMCs under basic conditions. Inflammatory cytokine IL-1ß promotes Ox-LDL uptake by HMCs. Furthermore, IL-1ß promotes the mRNA and protein expression of LOX-1, a specific receptor of Ox-LDL, suggesting that the enhancement of Ox-LDL uptake may be mediated by LOX-1 pathway. Anti-LOX-1 therapy may be a promising option for treatment of glomerulosclerosis.

3.
Artigo em Inglês | MEDLINE | ID: mdl-32439513

RESUMO

As a NAD+-dependent deacetylase, SIRT1 is widely involved in apoptosis and cellular inflammation via multiple pathways such as p53, NF-кB and STAT. More and more studies have shown that p53 is the first non-histone deacetylation target of SIRT1. SIRT1-p53 axis thus plays an important role in mammalian cells. IRF9 is an important member of interferon regulator factor family and performs an important role in innate immunity against foreign virus invasion. More importantly, human IRF9 can suppress the SIRT1-p53 axis. However, the functions and relationship between IRF9 and SIRT1-p53 axis are rarely studied in fish. To this end, we made a preliminary research on the functions of grass carp (Ctenopharyngodon idella) IRF9, SIRT1 and p53 in apoptosis and innate immunity. Firstly, we cloned and identified the ORF of SIRT1 (named CiSIRT1, MN125614) from C. idella and demonstrated that CiIRF9 promoted apoptosis, while CiSIRT1 inhibited apoptosis by flow cytometry and TUNEL experiments. Next, we found the interaction between CiSIRT1 and Cip53 in vivo by co-immunoprecipitation experiments. Moreover, the colocalization analysis also showed CiSIRT1 and Cip53 were mainly distributed in nucleus. Thirdly, we got a conclusion that CiIRF9 can repress the expression of CiSIRT1, implying that CiIRF9 regulates CiSIRT1-p53 axis. Finally, CiSIRT1 mRNA level was significantly up-regulated and the expression reached the highest level at 24 h post poly (I:C) stimulation in CIK cells. So, CiSIRT1 may exert an important function in innate immunity. Furthermore, we found CiSIRT1 down-regulated the expression of CiIFN1. In summary, CiIRF9 promotes apoptosis and innate immunity by inhibiting SIRT1-p53 axis. These findings will provide a new theoretical basis for the research on teleost innate immunity.

4.
QJM ; 2020 Apr 10.
Artigo em Inglês | MEDLINE | ID: mdl-32275753

RESUMO

BACKGROUND: Lungs from patients with coronavirus disease 2019 (COVID-19) have shown typical signs of acute respiratory distress syndrome (ARDS), formation of hyaline membrane mainly composed of fibrin, and 'ground-glass' opacity. Previously, we showed plasminogen itself is a key regulator in fibrin degradation, wound healing and infection. AIM: We aimed to investigate whether plasminogen can improve lung lesions and hypoxemia of COVID-19. DESIGN: Thirteen clinically moderate, severe or critical COVID-19 patients were treated with atomization inhalation of freeze-dried plasminogen. METHODS: Levels of their lung lesions, oxygen saturation and heart rates were compared before and after treatment by CT scanning images and patient monitor. RESULTS: After plasminogen inhalation, conditions of lung lesions in 5 clinically moderate patients have quickly improved, shown as the decreased range and density of 'ground glass' opacity. Improvements of oxygen saturation were observed in 6 clinically severe patients. In the 2 patients with critical conditions, the oxygen levels have significantly increased from 79-82% to 91% just about 1 hour after the first inhalation. In 8 of 13 patients the heart rates had slowed down. For the 5 clinically moderate patients, the difference is even statistically significant. Furthermore, a general relief of chest tightness was observed. CONCLUSION: Whereas it is reported that plasminogen is dramatically increased in adults with ARDS, this study suggests that additional plasminogen may be effective and efficient in treating lung lesions and hypoxemia during COVID-19 infections. Although further studies are needed, this study highlights a possible hope of efficiently combating this rapid epidemic emergency.

5.
Carbohydr Polym ; 233: 115831, 2020 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-32059884

RESUMO

Fibrillar gel of pepsin-solubilized collagen from tilapia skin was prepared by self-assembly in neutral phosphate buffer at 28 °C. Then effects of acidic polysaccharides, such as sodium alginate (SA), chondroitin sulfate (CS), and hyaluronic acid (HA), on the formation and properties of self-assembled fibrillar gel were investigated. SA and CS prolonged gelling time, whereas HA had no obvious effect. SA made fibril network denser, while CS and HA induced the presence of larger ordered structures. All the acidic polysaccharides broadened the D-periodicity of fibrils. SA and HA increased the maximum mechanical strength of gel to 39.64 and 34.49 kN/m2, respectively, significantly higher than that of pure collagen gel (14.53 kN/m2), while that only 17.20 kN/m2 after CS introduced. HA had no evident effect on enzymatic resistance, while SA and CS decreased. Therefore, tilapia skin collagen with HA has a higher potential as a biomaterial than that with CS or SA.

6.
BMC Cardiovasc Disord ; 20(1): 40, 2020 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-32000667

RESUMO

BACKGROUND: Endothelial dysfunction may play a key role in non-obstructive coronary artery atherosclerosis. Our study aimed to evaluate the vascular endothelial function and its influencing factors in patients with non-obstructive coronary artery atherosclerosis. METHODS: A total of 131 consecutive patients with non-obstructive coronary artery atherosclerosis were enrolled. Flow-mediated dilatation (FMD) was measured at baseline and 1-year follow-up. Endothelial progenitor cells (EPCs) were counted by staining the fasting venous blood with antibodies against CD34 and vascular endothelial growth factor receptor 2. RESULTS: Systolic blood pressure, pulse pressure and the levels of HbA1c in participants with baseline FMD < 6% (n = 65) were significantly higher than those with baseline FMD ≥ 6% (n = 66). Baseline FMD was negatively associated with EPC counts (r = - 0.199, P < 0.05) and systolic blood pressure (r = - 0.315, P < 0.01). The 1-year FMD was significantly increased compared to the baseline FMD [(9.31 ± 5.62) % vs (7.31 ± 5.26) %, P < 0.001]. Independent predictors of FMD improvement included elevated EPC counts (OR = 1.104, 95% CI: 1.047-1.165, P < 0.001) and decreased levels of serum creatinine (OR = 0.915, 95% CI: 0.843-0.993, P = 0.034). CONCLUSIONS: Family history of premature cardiovascular diseases, hypertension, elevated systolic pressure, and HbA1c > 6.5% are independent risk factors for endothelial dysfunction in non-obstructive atherosclerotic patients. Elevated peripheral blood EPC counts and decreased levels of serum creatinine are independent predictors of endothelial function improvement.

7.
Food Funct ; 11(3): 2173-2185, 2020 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-32083631

RESUMO

The immunomodulatory effects of R-phycoerythrin (R-PE) from Porphyra haitanensis were investigated by a hydrocortisone (HC)-induced immunosuppressive model in the present research. The results showed that R-PE had immunomodulatory potential, such as increasing spleen and thymus indexes, ameliorating spleen morphology, enhancing the phagocytic capability of macrophages, promoting spleen T lymphocyte proliferation, and strengthening serum hemolysin formation. Furthermore, R-PE elevated the CD4+/CD8+ ratio and up-regulated the levels of cytokines for Th1, Th2, and Th17 and the mRNA expression of transcription factors T-bet, GATA3, and RORγt. Besides, the levels of cytokines for Treg and transcription factor Foxp3 were down-regulated. The levels of T-bet/GATA3 and RORγt/Foxp3 were increased. R-PE activated the phosphorylation of NF-κB through the TLR4-mediated differentiation pathway. These findings suggested that R-PE exerted immunomodulatory activities in the innate and adaptive immune systems through TLR4/NF-κB mediated CD4+ T cell activation and differentiation, which facilitated the further application of R-PE as an immunomodulator.

8.
Int J Biol Macromol ; 148: 510-517, 2020 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-31958560

RESUMO

This study investigated the extraction of polysaccharide from okra pulp (POP), its chemical components, and rheological properties. The results showed that the optimal extraction temperature, time, and the ration of water to material were 20-30 °C, 30 mins, and 150, respectively, giving a maximum yield of 29.4%. The POP extracted under the mild condition showed different properties. The molecular weight of POP varied from 6436 kDa to 7432 kDa. The GalA/Rha and Gal/Rha in POP suggests the domain of rhamnogalacturonan I with long galactose side chains. The POP presented pseudoplastic shear-thinning behavior, which can be described by the Ostward-DeWaele model. The apparent viscosity of POP decreased with temperature rising from 25 °C to 80 °C. In addition, sucrose, CaCl2, and NaCl led to the reduction of its apparent viscosity which was more sensitive to Ca2+ than to Na+ and sucrose. A closed hysteresis loop was obtained when the POP concentration reached to 6 g/L. The POP showed an elastic behavior (G' > G″) at concentration of 6 g/L, while it showed predominantly viscous response (G' < G″) at concentration of 2 and 4 g/L, over a wide range of frequencies (0.1-10 Hz). These results are potentially useful for the application of POP in food processing.

9.
J Control Release ; 317: 232-245, 2020 Jan 10.
Artigo em Inglês | MEDLINE | ID: mdl-31783048

RESUMO

The success in the application of nanomedicines for tumor therapy is largely dependent on the development of efficient tumor targeting, specific and effective drug delivery systems. Here, through a simple synthetic process, we developed a type of novel glucose transporter 1 (GLUT1)-targeting, tumor microenvironment responsive and near infrared irradiation (NIR) induced cytosolic drug delivery nanoparticles (NPs). Our design was based on polydopamine (PDA) NPs as the photothermal agent and drug delivery carrier, glucosyl functional ligands as the GLUT1 targeting agents, and the conjugation of anticancer drug bortezomib (BTZ) to the catechol groups of PDA NPs in a pH-dependent manner. The in vitro and in vivo studies demonstrated that the functionalized PDA NPs could efficiently accumulate in tumor site and localize in subcellular endo/lysosomes of tumor cells, and they could respond to tumor microenvironment and endo/lysosomal pH as well as NIR to promote the robust release of BTZ. Furthermore, the functionalized PDA NPs were first demonstrated to overcome the endo/lysosomal barrier for the enhanced cytosolic BTZ drug delivery through NIR-triggered endo/lysosomal release, achieving the integration of NIR-triggered photothermal effect and chemotherapy for synergistic tumor ablation. The significant suppression and even complete regression of 4 T1 tumor was observed in mice given only single treatment. Therefore, the GLUT1-targeting, pH and photothermal responsive drug delivery NPs show a great potential for broadly applicable chemo-photothermal tumor therapy.

10.
Dev Comp Immunol ; 102: 103475, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31437525

RESUMO

In mammals, interferon regulatory factor 5 (IRF5) can be activated by tumor necrosis factor receptor-associated factor 6 (TRAF6). Upon activation, IRF5 translocates into the nucleus, where it binds to IFN promoter and up-regulates IFN expression. However, there are few reports on the molecular mechanism by which TRAF6 up-regulates IFN expression in fish. In this study, we explored how Grass carp (Ctenopharyngodon idellus) TRAF6 initiated innate immunity by activating IRF5. We found that CiTRAF6, CiIRF5 and CiIFN1 were all significantly up-regulated in LPS-stimulated CIK cells and the expression of CiTRAF6 was earlier than the expressions of CiIRF5 and CiIFN1. These findings suggested that CiIFN1 expression might be induced by CiTRAF6 in fish. CiIFN1 expression, CiIFN1 promoter activity and CO cells viability were all significantly up-regulated in the overexpression experiments, but they were significantly down-regulated in the gene silencing experiments. This indicated that CiTRAF6, along with CiIRF5, regulated CiIFN1 expression. The localization analysis found that both CiTRAF6 and CiIRF5 located in the cytoplasm. Following LPS stimulation, CiIRF5 was observed to translocate to the nucleus. GST-pull down and co-IP experiments revealed that CiTRAF6 interacted with CiIRF5. The colocalization analysis also showed that CiTRAF6 bound with CiIRF5 in the cytoplasm. Overexpression of CiTRAF6 increased the endogenous CiIRF5, promoted its ubiquitination and nuclear translocation. In conclusion, CiTRAF6 bound to CiIRF5 in the cytoplasm, and then activated CiIRF5, resulting in up-regulating the expression of CiIFN1.

11.
J Mater Chem B ; 7(38): 5848-5860, 2019 Oct 14.
Artigo em Inglês | MEDLINE | ID: mdl-31508651

RESUMO

Reducing surgical incision for large area subcutaneous defect filling and repair is a great challenge in the biomedical field, especially for plastic surgery. In this study, a novel hydroxyethyl cellulose/soy protein isolate (HEC/SPI) composite sponge (EHSS) with a fluid responsive shape memory property was constructed, whose thickness could be controlled by hot-pressing conditions to reduce the required surgical incision greatly. Effects of the main factors such as pressure, temperature and hot-pressing cycles on the recovery degree of EHSS were investigated systematically. The structure and physical properties of the sponges were characterized by FTIR spectroscopy, XRD, SEM etc. The results showed that EHSS could be pressed into thin disks with much smaller thickness, and the thickness retention ratio and recovery ratio were affected by hot-pressing conditions such as pressure and temperature. Especially, EHSS could be hot-pressed into a dense thin disk (EHSS-PT-130) at 130 °C with the pressure of 30 MPa, which could quickly recover its original shape by soaking in hydrophilic fluids. EHSS-PT-130 also exhibited good hydrophilicity, cytocompatibility, histocompatibility and in vivo biodegradability. Compared with the original EHSS, in vivo shape memory EHSS-PT-130 required much smaller surgical incision to reach the same repair effect and no need of extra sterilization, showing potential application for subcutaneous defect filling and repair.

12.
Appl Microbiol Biotechnol ; 103(19): 7997-8008, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31414160

RESUMO

The synthesis of rhamnosylated compounds has gained great importance since these compounds have potential therapeutic applications. The enzymatic approaches for glycosylation of bioactive molecules have been well developed; however, the enzymatic rhamnosylation has been largely hindered by lacking of the glycosyl donor for rhamnosyltransferases. Here, we employed an α-L-rhamnosidase from Alternaria sp. L1 (RhaL1) to perform one-step rhamnosylation of anticancer drugs, including 2'-deoxy-5-fluorouridine (FUDR), cytosine arabinoside (Ara C), and hydroxyurea (Hydrea). The key synthesis conditions including substrate concentrations and reaction time were carefully optimized, and the maximum yields of each rhamnosylated drugs were 57.7 mmol for rhamnosylated Ara C, 68.6 mmol for rhamnosylated Hydrea, and 42.2 mmol for rhamnosylated FUDR. It is worth pointing out that these rhamnosylated drugs exhibit little cytotoxic effects on cancer cells, but could efficiently restore cytotoxic activity when incubated with exogenous α-L-rhamnosidase, suggesting their potential applications in the enzyme-activated prodrug system. To evaluate the cancer-targeting ability of rhamnose moiety, the rhamnose-conjugated fluorescence dye rhodamine B (Rha-RhB) was constructed. The fluorescence probe Rha-RhB displayed much higher cell affinity and cellular internalization rate of oral cancer cell KB and breast cancer cell MDA-MB-231 than that of the normal epithelial cells MCF 10A, suggesting that the rhamnose moiety could mediate the specific internalization of rhamnosylated compounds into cancer cells, which greatly facilitated their applications for cancer-targeting drug delivery.


Assuntos
Alternaria/enzimologia , Antineoplásicos/metabolismo , Glicosídeo Hidrolases/metabolismo , Terapia de Alvo Molecular/métodos , Neoplasias/tratamento farmacológico , Pró-Fármacos/metabolismo , Ramnose/metabolismo , Antineoplásicos/farmacologia , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Citarabina/metabolismo , Citarabina/farmacologia , Floxuridina/metabolismo , Floxuridina/farmacologia , Humanos , Hidroxiureia/metabolismo , Hidroxiureia/farmacologia , Pró-Fármacos/farmacologia
13.
Biosens Bioelectron ; 142: 111547, 2019 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-31387025

RESUMO

Glucose sensing is of vital importance due to the growing number of diabetes. In this study, we developed a visual detecting approach for glucose sensing based on a smart hydrogel system, by assembling of a photo-crosslinkable hydrogel and a pH-responsive nanogel, respectively. The hydrogel system showed fast response and high sensitivity to glucose in the physiological ranges, and enabled a visual detection of glucose both in vitro in glucose solutions and in vivo in diabetic mouse models. In normoglycemic state, the hydrogels showed large swelling, resulting in a large shape but with weak color or fluorescence intensity of the hydrogels. In hyperglycemic state, the hydrogels exhibited less swelling, resulting in a small shape but with strong color or fluorescence intensity of the hydrogels. Based on the observation of the size change and intensity change of the hydrogels, we can visual the glucose levels by either colorimetry or fluorescence imaging. This hydrogel system provides a novel means for visual detection of glucose. Our study broadens the current applications of hydrogels, extending their potentials in clinical diagnosis of diabetes or glucose-related analysis.


Assuntos
Acrilatos/química , Técnicas Biossensoriais/métodos , Glicemia/análise , Carboximetilcelulose Sódica/química , Gelatina/química , Hidrogéis/química , Animais , Colorimetria/métodos , Preparações de Ação Retardada/química , Diabetes Mellitus Experimental/sangue , Glucose/análise , Concentração de Íons de Hidrogênio , Luz , Masculino , Camundongos Endogâmicos BALB C , Camundongos Nus
14.
Zhonghua Wei Zhong Bing Ji Jiu Yi Xue ; 31(6): 665-668, 2019 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-31315719

RESUMO

OBJECTIVE: Severe community acquired pneumonia (SCAP) has raised substantial concern due to high morbidity and mortality. SCAP is usually associated with multiple organ dysfunction syndrome (MODS) and sepsis. It is suggested that organ function and coagulation/fibrinolysis improvement, organ injury reduction and organ repair facilitation are highly required besides microbiology detection and optimization of antibiotics strategy on treatment of SCAP. Neutralization antibody, steroid, immunmodulatory therapy, and Chinese medicine etc. contribute to organ function improvement from different perspective. The unique profile of multiple target and personalized therapy of Chinese medicine may become a promising new option for SCAP and MODS. The combination of Western and Chinese Medicine shed new light on SCAP treatment.


Assuntos
Infecções Comunitárias Adquiridas/terapia , Pneumonia/terapia , Antibacterianos , Humanos , Insuficiência de Múltiplos Órgãos , Sepse , Índice de Gravidade de Doença
15.
BMC Gastroenterol ; 19(1): 86, 2019 Jun 13.
Artigo em Inglês | MEDLINE | ID: mdl-31195990

RESUMO

BACKGROUND: Hepatitis C virus (HCV) infection is commonly associated with a disturbance of glucose metabolism. However, there have been conflicting reports on whether the clearance of the HCV may be followed by changes of serum blood glucose and insulin resistance. The aim of the present study was to evaluate the impact of HCV and antiviral treatment on serum glucose levels and other glucose metabolism parameters. METHODS: A retrospective analysis of 306 HCV-infected patients was performed. Fasting serum blood glucose (FBG) levels in these patients were compared with that of 325 healthy individuals. Serum parameters of glucose metabolism were measured in 183 patients with chronic hepatitis C at baseline, at the end of interferon α-2b plus ribavirin treatment, and at 24 weeks post-treatment. RESULTS: Patients with HCV infection had significantly higher FBG level than healthy controls (5.57 ± 0.74 vs. 5.11 ± 0.83 mmol/l, P < 0.001). After antiviral treatment, we found a significant reduction in FBG levels regardless of the outcome of treatment. However, after stopping treatment the serum FBG levels were significantly elevated in the sustained virological response (SVR) and non-responder groups, and maintained high level until week 24 post-treatment. In both groups, the levels of serum FBG after 24 weeks post-treatment were still lower than pre-treatment levels. In sustained responders, fasting insulin (P = 0.007), C-peptide (P < 0.001) and HOMA-IR (P < 0.001) significantly decreased, and the insulin sensitivity index (ISI) increased (P < 0.001) at the end of the treatment comparing with pre-treatment levels, while no significant difference was observed in non-responder group. HOMA-ß values were increased in both groups at the end of treatment (both P < 0.001). CONCLUSION: The total serum FBG level in HCV infected patients was higher than that in healthy controls. Clearance of HCV was associated with reduced glucose and improved insulin resistance.


Assuntos
Antivirais/administração & dosagem , Glicemia/análise , Hepacivirus , Hepatite C Crônica/sangue , Resistência à Insulina , Adulto , Quimioterapia Combinada , Jejum/sangue , Feminino , Hepatite C Crônica/tratamento farmacológico , Hepatite C Crônica/virologia , Humanos , Insulina/sangue , Interferon-alfa/administração & dosagem , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Ribavirina/administração & dosagem , Resposta Viral Sustentada
16.
Dev Comp Immunol ; 99: 103410, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31175887

RESUMO

As a member of NDR protein kinase family and a novel protein kinase of Hippo signal pathway, Serine/threonine kinase 38 (STK38) plays a very significant role in the innate immune. In mammals, STK38 performs its function by combining with GSK3ß. Nowadays, there are few reports of STK38 in fish. In order to explain the function of fish STK38 in the innate immunity, we cloned the ORF of grass carp (Ctenopharyngodon idella) STK38 (CiSTK38) and the related kinase GSK3ß (CiGSK3ß). Phylogenetic trees revealed that CiSTK38 and CiGSK3ß evolved closer kinship with sinocyclocheilus grahami STK38 and siniperca chuatsi GSK3ß respectively. CiSTK38 and CiGSK3ß can respond to the intradermal injection of poly (I:C) in grass carp different tissues and the transfection of poly I:C in CIK cells. Subcellular localization revealed the CiGSK3ß were broadly distributed through the cytoplasm, whereas CiSTK38 were observed both in cytoplasm and nucleus. However, when they were co-transferred into cells, the two proteins were found to aggregate in the nucleus. GST-pulldown and co-immunoprecipitation analysis revealed that CiSTK38 can physically interact with CiGSK3ß. Phos-tag PAGE illustrated CiSTK38 can decrease the phosphorylation and auto-phosphorylation level of CiGSK3ß at Ser9 and at Tyr216. To investigate the functional correlation between CiSTK38 and CiGSK3ß, we overexpressed CiSTK38 and CiGSK3ß in CIK cells and found that they can up-regulate the expression of IFN I. In short, we demonstrated that CiSTK38 can confer CiGSK3ß kinase activity by reducing its phosphorylation level. Result from this study strongly suggested that the anti-viral immune effects elicited by poly (I:C) in part were mediated through activation of CiGSK3ß. The findings provided scientific basis for the anti-viral immune mechanism of STK38 and GSK3ß in fish.


Assuntos
Carpas/imunologia , Proteínas de Peixes/metabolismo , Regulação da Expressão Gênica , Glicogênio Sintase Quinase 3 beta/metabolismo , Interferon Tipo I/genética , Proteínas Serina-Treonina Quinases/metabolismo , Animais , Carpas/classificação , Carpas/genética , Linhagem Celular , Núcleo Celular/metabolismo , Citoplasma/metabolismo , Proteínas de Peixes/genética , Técnicas de Silenciamento de Genes , Glicogênio Sintase Quinase 3 beta/genética , Imunidade Inata/genética , Fosforilação , Filogenia , Poli I-C/imunologia , Ligação Proteica , Proteínas Serina-Treonina Quinases/genética
17.
J Econ Entomol ; 112(5): 2085-2093, 2019 09 23.
Artigo em Inglês | MEDLINE | ID: mdl-31115452

RESUMO

Western flower thrips, Frankliniella occidentalis (Pergande), is a destructive insect pest in greenhouse production systems. Therefore, integrating the entomopathogenic fungus, Beauveria bassiana (Balsamo) Vuillemin, with the soil-dwelling rove beetle, Dalotia coriaria (Kraatz), targeting different aboveground and belowground life stages may help effectively manage western flower thrips populations. Two greenhouse experiments were conducted evaluating five treatments: 1) insecticides (spinosad, pyridalyl, chlorfenapyr, and abamectin), 2) B. bassiana, 3) D. coriaria, 4) B. bassiana and D. coriaria combination, and 5) water control. The estimated mean number of western flower thrips adults captured on yellow sticky cards was significantly lower for the insecticide treatment (mean range: 0-46 western flower thrips adults per yellow sticky card) than the B. bassiana and D. coriaria combination (0.3-105.1 western flower thrips per yellow card) over 8 wk. There were no significant differences in the final foliar damage ratings of chrysanthemum, Dendranthema × grandiflorum (Ramat.) Kitam., plants among the five treatments in experiment 1, but there were significant differences in experiment 2. In experiment 2, chrysanthemum plants across all treatments were not marketable due to western flower thrips feeding damage. Therefore, using B. bassiana and D. coriaria early in production should suppress population growth by targeting both foliar-feeding and soil-dwelling life stages of western flower thrips simultaneously.


Assuntos
Besouros , Hypocreales , Inseticidas , Tisanópteros , Animais , Flores , Insetos
18.
J Cell Physiol ; 234(10): 18906-18916, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30953350

RESUMO

Bone marrow mesenchymal stem cells (BMSC) can ameliorate ischemic injury of various tissues. However, the molecular mechanisms involved remain to be clarified. In this study, we intend to investigate the effects of BMSC-derived conditioned medium (BMSC-CM) on hypoxia/reoxygenation (H/R)-induced injury of H9c2 myocardial cells, and the potential mechanisms. Cell injury was determined through level of cell viability, lactate dehydrogenase (LDH) release, total intracellular reactive oxygen species (ROS), mitochondrial membrane potential (Δψm), and cell apoptosis. Autophagic activity of cells was detected through levels of the autophagy-associated proteins and autophagic flux. Results showed that BMSC-CM alleviated H/R-induced injury in H9c2 cells, as demonstrated by increased cell viability and Δψm, decreased ROS production, LDH release, and cell apoptosis. Furthermore, the H/R treatment induced a decrease in autophagic activity and an increase in Notch2 signaling activation in H9c2 cells. In the presence of BMSC-CM, the autophagic activity impaired by the H/R treatment was upregulated with decreased phosphorylation of mTOR, and the activation of Notch2 signaling was downregulated. These effects of BMSC-CM could be replicated by Notch signaling inhibitor. In contrast, inhibitors of cell autophagy including chloroquine (CQ) and 3-methyladenine, diminished the protective effects of BMSC-CM. Taken together results, our study showed that BMSC-CM could protect H9c2 cells from H/R-induced injury potentially through regulating Notch2/mTOR/autophagy signaling. These findings may provide a novel insight into the mechanisms of BMSC-CM in therapy of myocardial ischemia/reperfusion injury as well as other ischemic diseases.

19.
Int J Mol Sci ; 20(8)2019 Apr 12.
Artigo em Inglês | MEDLINE | ID: mdl-31013701

RESUMO

Phytophthora root rot (PRR) causes serious annual soybean yield losses worldwide. The most effective method to prevent PRR involves growing cultivars that possess genes conferring resistance to Phytophthora sojae (Rps). In this study, QTL-sequencing combined with genetic mapping was used to identify RpsX in soybean cultivar Xiu94-11 resistance to all P. sojae isolates tested, exhibiting broad-spectrum PRR resistance. Subsequent analysis revealed RpsX was located in the 242-kb genomic region spanning the RpsQ locus. However, a phylogenetic investigation indicated Xiu94-11 carrying RpsX is distantly related to the cultivars containing RpsQ, implying RpsX and RpsQ have different origins. An examination of candidate genes revealed RpsX and RpsQ share common nonsynonymous SNP and a 144-bp insertion in the Glyma.03g027200 sequence encoding a leucine-rich repeat (LRR) region. Glyma.03g027200 was considered to be the likely candidate gene of RpsQ and RpsX. Sequence analyses confirmed that the 144-bp insertion caused by an unequal exchange resulted in two additional LRR-encoding fragments in the candidate gene. A marker developed based on the 144-bp insertion was used to analyze the genetic population and germplasm, and proved to be useful for identifying the RpsX and RpsQ alleles. This study implies that the number of LRR units in the LRR domain may be important for PRR resistance in soybean.


Assuntos
Resistência à Doença/genética , Interações Hospedeiro-Parasita/genética , Phytophthora , Doenças das Plantas/genética , Doenças das Plantas/parasitologia , Soja/genética , Soja/parasitologia , Mapeamento Cromossômico , Cromossomos de Plantas , Sequência Conservada , Genes de Plantas , Ligação Genética , Loci Gênicos , Genótipo , Sequenciamento de Nucleotídeos em Larga Escala , Filogenia , Phytophthora/isolamento & purificação , Polimorfismo de Nucleotídeo Único , Soja/classificação
20.
Zhonghua Wei Zhong Bing Ji Jiu Yi Xue ; 31(2): 135-138, 2019 Feb.
Artigo em Chinês | MEDLINE | ID: mdl-30827296

RESUMO

OBJECTIVE: The "three certificates and three methods" is a combination of Chinese and Western medicine for the treatment of sepsis proposed by Professor Wang Jinda, a pioneer in Chinese emergency medicine. The "Wei Qi Ying Xue" syndrome differentiation is a dialectical theory proposed by Ye Tianshi, a famous doctor in the Qing Dynasty, for the treatment of warm diseases. The "three certificates and three methods" theory of "toxin-heat syndrome" (inflammation storm) to the "blood stasis syndrome" (coagulation storm) conversion mainly focus on the "Wei Qi Ying Xue" syndrome differentiation of the "Wei, Qi syndrome" to "Ying, Xue syndrome", and poison. The conversion of "toxin-heat syndrome" to "blood stasis syndrome" is a watershed in the progression of common infections to sepsis. The timely application of Xuebijing injection can cut off this transformation, which is one of the reasons why Xuebijing injection can treat sepsis.


Assuntos
Sepse/diagnóstico , Diagnóstico Diferencial , Humanos , Síndrome
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