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Cerebral ischemic preconditioning (CIP) has been shown to improve brain ischemic tolerance against subsequent lethal ischemia. Reactive astrocytes play important roles in cerebral ischemia-reperfusion. Recent studies have shown that reactive astrocytes can be polarized into neurotoxic A1 phenotype (C3d) and neuroprotective A2 phenotype (S100A10). However, their role in CIP remains unclear. Here, we focused on the role of N-myc downstream-regulated gene 2 (NDRG2) in regulating the transformation of A1/A2 astrocytes and promoting to brain ischemic tolerance induced by CIP. A Sprague Dawley rat model of middle cerebral artery occlusion/reperfusion (MCAO/R) was used. Rats were divided into the following six groups: (1) sham group; (2) CIP group: left middle cerebral artery was blocked for 10 min; (3) MCAO/R group: left middle cerebral artery was blocked for 90 min; (4) CIP + MCAO/R group: CIP was performed 72 h before MCAO/R; (5) AAV-NDRG2 + CIP + MCAO/R group: adeno-associated virus (AAV) carrying NDRG2 was administered 14 days before CIP + MCAO/R; (6) AAV-Ctrl + CIP + MCAO/R group: empty control group. The rats were subjected to neurological evaluation 24 h after the above treatments, and then were sacrificed for 2, 3, 5-triphenyltetraolium chloride staining, thionin staining, immunofluorescence and western blot analysis. In CIP + MCAO/R group, the neurological deficit scores decreased, infarct volume reduced, and neuronal density increased compared with MCAO/R group. Notably, CIP significantly increased S100A10 expression and the number of S100A10+/GFAP+ cells, and also increased NDRG2 expression. MCAO/R significantly decreased S100A10 expression and the number of S100A10+/GFAP+ cells yet increased C3d expression and the number of C3d+/GFAP+ cells and NDRG2 expression, and these trends were reversed by CIP + MCAO/R. Furthermore, over-expression of NDRG2 before CIP + MCAO/R, the C3d expression and the number of C3d+/GFAP+ cells increased, while S100A10 expression and the number of S100A10+/GFAP+ cells decreased. Meanwhile, over-expression of NDRG2 blocked the CIP-induced brain ischemic tolerance. Taken together, these results suggest that CIP exerts neuroprotective effects against ischemic injury by suppressing A1 astrocyte polarization and promoting A2 astrocyte polarization via inhibiting NDRG2 expression.
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Astrocitos , Isquemia Encefálica , Infarto de la Arteria Cerebral Media , Precondicionamiento Isquémico , Ratas Sprague-Dawley , Animales , Precondicionamiento Isquémico/métodos , Masculino , Astrocitos/metabolismo , Infarto de la Arteria Cerebral Media/metabolismo , Infarto de la Arteria Cerebral Media/patología , Isquemia Encefálica/metabolismo , Ratas , Proteínas del Tejido NerviosoRESUMEN
Cerebral ischemic preconditioning (CIP)-induced brain ischemic tolerance protects neurons from subsequent lethal ischemic insult. However, the specific mechanisms underlying CIP remain unclear. In the present study, we explored the hypothesis that peroxisome proliferator-activated receptor gamma (PPARγ) participates in the upregulation of Klotho during the induction of brain ischemic tolerance by CIP. First we investigated the expression of Klotho during the brain ischemic tolerance induced by CIP. Lethal ischemia significantly decreased Klotho expression from 6 h to 7 days, while CIP significantly increased Klotho expression from 12 h to 7 days in the hippocampal CA1 region. Inhibition of Klotho expression by its shRNA blocked the neuroprotection induced by CIP. These results indicate that Klotho participates in brain ischemic tolerance by CIP. Furthermore, we tested the role of PPARγ in regulating Klotho expression after CIP. CIP caused PPARγ protein translocation to the nucleus in neurons in the CA1 region of the hippocampus. Pretreatment with GW9962, a PPARγ inhibitor, significantly attenuated the upregulation of Klotho protein and blocked the brain ischemic tolerance induced by CIP. Taken together, it can be concluded that Klotho upregulation via PPARγ contributes to the induction of brain ischemic tolerance by CIP.
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Isquemia Encefálica , Precondicionamiento Isquémico , Animales , Ratas , Isquemia Encefálica/metabolismo , Región CA1 Hipocampal , Isquemia , PPAR gamma/metabolismo , Ratas Wistar , Regulación hacia ArribaRESUMEN
OBJECTIVES: To explore the clinical potential of multiparametric cardiac magnetic resonance (CMR) in evaluating myocardial inflammation in patients with exertional heat illness (EHI). METHODS: This prospective study enrolled 28 males with EHI (18 patients with exertional heat exhaustion (EHE) and 10 with exertional heat stroke (EHS)) and 18 age-matched male healthy controls (HC). All subjects underwent multiparametric CMR, and 9 patients had follow-up CMR measurements 3 months after recovery from EHI. CMR-derived left ventricular geometry, function, strain, native T1, extracellular volume (ECV), T2, T2*, and late gadolinium enhancement (LGE) were obtained and compared among different groups. RESULTS: Compared with HC, EHI patients showed increased global ECV, T2, and T2* values (22.6% ± 4.1 vs. 19.7% ± 1.7; 46.8 ms ± 3.4 vs. 45.1 ms ± 1.2; 25.5 ms ± 2.2 vs. 23.8 ms ± 1.7; all p < 0.05). Subgroup analysis showed that ECV was higher in the EHS patients than those in EHE and HC groups (24.7% ± 4.9 vs. 21.4% ± 3.2, 24.7% ± 4.9 vs. 19.7% ± 1.7; both p < 0.05). Repeated CMR measurements at 3 months after baseline CMR showed persistently higher ECV than HC (p = 0.042). CONCLUSIONS: With multiparametric CMR, EHI patients demonstrated increased global ECV, T2, and persistent myocardial inflammation at 3-month follow-up after EHI episode. Therefore, multiparametric CMR might be an effective method in evaluating myocardial inflammation in patients with EHI. CLINICAL RELEVANCE STATEMENT: This study showed persistent myocardial inflammation after an exertional heat illness (EHI) episode demonstrated by multiparametric CMR, which is a potential promising method to evaluate the severity of myocardial inflammation and guide return to work, play, or duty in EHI patients. KEY POINTS: ⢠EHI patients showed an increased global extracellular volume (ECV), late gadolinium enhancement, and T2 value, indicating myocardial edema and fibrosis. ⢠ECV was higher in the exertional heat stroke patients than exertional heat exhaustion and healthy control groups (24.7% ± 4.9 vs. 21.4% ± 3.2, 24.7% ± 4.9 vs. 19.7% ± 1.7; both p < 0.05). ⢠EHI patients showed persistent myocardial inflammation with higher ECV than healthy controls 3 months after index CMR (22.3% ± 2.4 vs. 19.7% ± 1.7, p = 0.042).
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Agotamiento por Calor , Golpe de Calor , Miocarditis , Humanos , Masculino , Medios de Contraste/farmacología , Estudios Prospectivos , Agotamiento por Calor/patología , Gadolinio , Función Ventricular Izquierda , Imagen por Resonancia Cinemagnética , Estudios de Casos y Controles , Miocardio/patología , Espectroscopía de Resonancia Magnética , Golpe de Calor/complicaciones , Golpe de Calor/diagnóstico por imagen , Golpe de Calor/patología , Inflamación/diagnóstico por imagen , Inflamación/patología , Valor Predictivo de las PruebasRESUMEN
BACKGROUNDS: Due to the unexpected side effects of the iodinated contrast agents, novel contrast agents for X-ray computed tomography (CT) imaging are urgently needed. Nanoparticles made by heavy metal elements are often employed, such as gold and bismuth. These nanoparticles have the advantages of long in vivo circulation time and tumor targeted ability. However, due to the long residence time in vivo, these nanoparticles may bring unexpected toxicity and, the preparation methods of these nanoparticles are complicated and time-consuming. METHODS: In this investigation, a small molecular bismuth chelate using diethylenetriaminepentaacetic acid (DPTA) as the chelating agent was proposed to be an ideal CT contrast agent. RESULTS: The preparation method is easy and cost-effective. Moreover, the bismuth agent show better CT imaging for kidney than iohexol in the aspect of improved CT values. Up to 500 µM, the bismuth agent show negligible toxicity to L02 cells and negligible hemolysis. And, the bismuth agent did not induce detectable morphology changes to the main organs of the mice after intravenously repeated administration at a high dose of 250 mg/kg. The pharmacokinetics of the bismuth agent follows the first-order elimination kinetics and, it has a short half-life time of 0.602 h. The rapid clearance from the body promised its excellent biocompatibility. CONCLUSIONS: This bismuth agent may serve as a potential candidate for developing novel contrast agent for CT imaging in clinical applications.
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Bismuto , Medios de Contraste , Tomografía Computarizada por Rayos X/métodos , Animales , Bismuto/química , Bismuto/farmacocinética , Bismuto/toxicidad , Medios de Contraste/química , Medios de Contraste/farmacocinética , Medios de Contraste/toxicidad , Yohexol/química , Yohexol/farmacocinética , Riñón/diagnóstico por imagen , Riñón/metabolismo , Nanopartículas del Metal/química , Nanopartículas del Metal/toxicidad , Ratones , Ácido Pentético/química , Ácido Pentético/farmacocinética , Distribución Tisular , Imagen de Cuerpo EnteroRESUMEN
Glial glutamate transporter 1 (GLT-1) plays a vital role in the induction of brain ischemic tolerance (BIT) by ischemic preconditioning (IPC). However, the mechanism still needs to be further explained. The aim of this study was to investigate whether peroxisome proliferator-activated receptor gamma (PPARγ) participates in regulating GLT-1 during the acquisition of BIT induced by IPC. Initially, cerebral IPC induced BIT and enhanced PPARγ and GLT-1 expression in the CA1 hippocampus in rats. The ratio of nuclear/cytoplasmic PPARγ was also increased. At the same time, the up-regulation of PPARγ expression in astrocytes in the CA1 hippocampus was revealed by double immunofluorescence for PPARγ and glial fibrillary acidic protein. Then, the mechanism by which PPARγ regulates GLT-1 was studied in rat cortical astrocyte-neuron cocultures. We found that IPC [45 min of oxygen glucose deprivation (OGD)] protected neuronal survival after lethal OGD (4 h of OGD), which usually leads to neuronal death. The activation of PPARγ occurred earlier than the up-regulation of GLT-1 in astrocytes after IPC, as determined by western blot and immunofluorescence. Moreover, the preadministration of the PPARγ antagonist T0070907 or PPARγ siRNA significantly attenuated GLT-1 up-regulation and the neuroprotective effects induced by IPC in vitro. Finally, the effect of the PPARγ antagonist on GLT-1 expression and BIT was verified in vivo. We observed that the preadministration of T0070907 by intracerebroventricular injection dose-dependently attenuated the up-regulation of GLT-1 and BIT induced by cerebral IPC in rats. In conclusion, PPARγ participates in regulating GLT-1 during the acquisition of BIT induced by IPC. Cover Image for this issue: doi: 10.1111/jnc.14532. Open Science: This manuscript was awarded with the Open Materials Badge For more information see: https://cos.io/our-services/open-science-badges/.
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Encéfalo/irrigación sanguínea , Encéfalo/metabolismo , Transportador 2 de Aminoácidos Excitadores/metabolismo , Precondicionamiento Isquémico , PPAR gamma/metabolismo , Animales , Isquemia Encefálica/metabolismo , Técnicas In Vitro , Masculino , Neuroglía/metabolismo , Ratas , Ratas WistarRESUMEN
Cancer stem cells (CSCs) play critical roles in cancer, making them important targets for new diagnostic and therapeutic approaches. Since CSCs are heterogeneous and not abundant in tumors, and few specific markers for these cells currently exist, new methods to isolate and characterize them are required. To address this issue, we developed a new label-free methodology to isolate, enrich, and identify CSCs from an heterogeneous tumor cell subpopulation using a cell sorting method (sedimentation field flow fractionation, SdFFF) and a biosensor as a detector. Enrichment was optimized using an original protocol and U87-MG glioblastoma cells cultured in a normal (N) or defined (D) medium (± fetal bovine serum, FBS) under normoxic (N, pO2 = 20%) or hypoxic (H, pO2 < 2%) conditions to obtain four cell populations: NN, NH, DN, and DH. After elution of CSCs via SdFFF using the hyperlayer mode (inertial elution mode for micrometer-sized species), we isolated eight subpopulations with distinct CSC contents based on phenotypical and functional properties, ranging from NN F1 with a lower CSC content to DH F3 with a higher CSC content. Reflecting biological differences, the intrinsic intracellular dielectric permittivity increased from NN to DH conditions. The largest difference in electromagnetic signature was observed between NN F1 and DH F3, in which the CSC content was lowest and highest, respectively. The results demonstrate that microwave dielectric spectroscopy can be used to reliably and efficiently distinguish stem cell characteristics. This new instrumental and methodological approach is an important innovation that allows both enrichment and detection of CSCs, opening the door to novel diagnostic and therapeutic approaches.
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Separación Celular/métodos , Fraccionamiento de Campo-Flujo/métodos , Glioblastoma/patología , Células Madre Neoplásicas/patología , Técnicas Biosensibles/instrumentación , Técnicas Biosensibles/métodos , Línea Celular Tumoral , Movimiento Celular , Separación Celular/instrumentación , Diseño de Equipo , Fraccionamiento de Campo-Flujo/instrumentación , HumanosRESUMEN
OBJECTIVE: To determine the correlation between quantitative results of contrast enhanced ultrasound and clinical and laboratory index for synovium of patients with rheumatoid arthritis (RA). METHODS: Thirty five RA patients participated in this study, with ten normal adults serving as controls. They were given a score of DAS28 and underwent laboratory tests, including erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP). Contrast-enhanced ultrasound examinations were performed to detect thickness of synovium (2D and Power Dopple). RESULTS: Peak intensity (PI) and area under the curve (AUC) were positively correlated with ESR, CRP and DAS28 score, respectively (P<0. 01). The rest of ultrasound parameters had no significant correlations with ESR, CRP and DAS28 score (P>0. 05). Synovium thickness identified by contrast enhanced ultrasound had significant correlations with ESR, CRP and DAS28 score (P < 0. 01). CONCLUSION: Synovium thickness determined by contrast enhanced ultrasound, PI and AUC can serve as an index estimating inflammatory activity of RA, and provide evidence for clinical diagnosis and treatment.
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Artritis Reumatoide/diagnóstico por imagen , Índice de Severidad de la Enfermedad , Membrana Sinovial/diagnóstico por imagen , Adulto , Área Bajo la Curva , Sedimentación Sanguínea , Proteína C-Reactiva/análisis , Estudios de Casos y Controles , Humanos , UltrasonografíaRESUMEN
OBJECTIVE: To determine the diagnostic value of ultrasound imaging in chronic gouty arthritis. METHODS: Ultrasound imaging of 32 patients with chronic gouty arthritis were compared with those of 36 patients with non-gouty joint diseases. A logistic model was obtained to establish the associations between ultrasound appereances and gouty arthritis. Receiver operator characteristic (ROC) curve was constructed to evaluate the performance of the logistic model. RESULTS: A double contour sign of articular cartilage, a snowstorm appearance of the joint fluid, and a hyperecho around the periarticular tendon had diagnostic value for chronic gouty arthritis. According to the logistic regression analysis, the accuracy of predicting chronic gouty arthritis was 95.59% (65/ 68), and the area under curve of ROC was 3.987 +/- 0.011 (P < 0.05). CONCLUSION: A double contour sign of articular cartilage, a snowstorm appearance of the joint fluid, and a hyperecho around the periarticular tendon have diagnostic value. The combination of these appearances can improve diagnosis of chronic gouty arthritis.
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Artritis Gotosa/diagnóstico por imagen , Cartílago Articular/patología , Líquido Sinovial , Enfermedad Crónica , Humanos , Modelos Logísticos , Curva ROC , UltrasonografíaRESUMEN
Our previous study has proved that the Klotho up-regulation participated in cerebral ischemic preconditioning (CIP)-induced brain ischemic tolerance. However, the exact neuroprotective mechanism of Klotho in CIP remains unclear. We explored the hypothesis that STAT4-mediated Klotho up-regulation contributes to the CIP-induced brain ischemic tolerance via inhibiting neuronal pyroptosis. Firstly, the expressions of pyroptosis-associated proteins (i.e., NLRP3, GSDMD, pro-caspase-1, and cleaved caspase-1) in hippocampal CA1 region were determined during the process of brain ischemic tolerance. We found the expression of pyroptosis-associated proteins was significantly up-regulated in the ischemic insult (II) group, and showed no significant changes in the CIP group. The expression level of each pyroptosis-associated proteins was lower in the CIP + II group than that in the II group. Inhibition of Klotho expression increased the expression of pyroptosis-associated proteins in the CIP + II group and blocked the CIP-induced brain ischemic tolerance. Injection of Klotho protein decreased the expression of pyroptosis-associated proteins in the II group, and protected neurons from ischemic injury. Secondly, the transcription factor STAT4 of Klotho was identified by bioinformatic analysis. Double luciferase reporter gene assay and chromatin immunoprecipitation assay showed STAT4 can bind to the site between nt - 881 and - 868 on the Klotho promoter region and positively regulates Klotho expression. Moreover, we found CIP significantly enhanced the expression of STAT4. Knockdown STAT4 suppressed Klotho up-regulation after CIP and blocked the CIP-induced brain ischemic tolerance. Collectively, it can be concluded that STAT4-mediated the up-regulation of Klotho contributed to the brain ischemic tolerance induced by CIP via inhibiting pyroptosis.
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Isquemia Encefálica , Precondicionamiento Isquémico , Ratas , Animales , Ratas Wistar , Regulación hacia Arriba , Piroptosis , Factor de Transcripción STAT4/metabolismo , Isquemia Encefálica/metabolismo , Región CA1 Hipocampal/metabolismo , Neuronas/metabolismo , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismoRESUMEN
The morbidity rate of ischemic stroke is increasing annually with the growing aging population in China. Astrocytes are ubiquitous glial cells in the brain and play a crucial role in supporting neuronal function and metabolism. Increasing evidence shows that the impairment or loss of astrocytes contributes to neuronal dysfunction during cerebral ischemic injury. The mitochondrion is increasingly recognized as a key player in regulating astrocyte function. Changes in astrocytic mitochondrial function appear to be closely linked to the homeostasis imbalance defects in glutamate metabolism, Ca2+ regulation, fatty acid metabolism, reactive oxygen species, inflammation, and copper regulation. Here, we discuss the role of astrocytic mitochondria in the pathogenesis of brain ischemic injury and their potential as a therapeutic target.
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Lesiones Encefálicas , Isquemia Encefálica , Humanos , Anciano , Astrocitos/metabolismo , Isquemia Encefálica/patología , Encéfalo/metabolismo , Lesiones Encefálicas/metabolismo , Mitocondrias/metabolismoRESUMEN
OBJECTIVE: To screen interleukin (IL)-1ß secretion-related membrane transporters by macrophage experiment in vitro and conventional knockout mice. METHODS: THP-1 cell line was differentiated to obtain human THP-1-derived macrophages, and the primary macrophages were obtained from human peripheral blood. FVB wild-type mice with the same sex and age were used as the controls of MRP1 knockout mice. The macrophages in abdominal cavity and bone marrow of mice were cultivated. The cells were treated with ABCC1/MRP1, ABCG2/BCRP, ABCB1/P-gp, OATP1B1, and MATE transporter inhibitors, then stimulated by lipopolysaccharide and adenosine triphosphate. The secretion level of IL-1ß was detected by ELISA, Western blot, and immunofluorescence. RESULTS: After inhibiting ABCC1/MRP1 transporter, the secretion of IL-1ß decreased significantly, while inhibition of the other 4 transporters had no effect. In animal experiment, the level of IL-1ß secreted by macrophages in bone marrow of MRP1 knockout mice was significantly lower than control group (P < 0.05). CONCLUSION: ABCC1/MRP1 transporter is a newly discovered IL-1ß secretion pathway, which is expected to become a new target for solving clinical problems such as cytokine release syndrome.
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Regulación hacia Abajo , Interleucina-1beta , Macrófagos , Ratones Noqueados , Proteínas Asociadas a Resistencia a Múltiples Medicamentos , Animales , Humanos , Ratones , Interleucina-1beta/metabolismo , Lipopolisacáridos , Macrófagos/metabolismo , Proteínas Asociadas a Resistencia a Múltiples Medicamentos/metabolismo , Células THP-1RESUMEN
BACKGROUND: Hepatic pseudolesions detected by helical computed tomography (CT) are not rare, but it is difficult to make a final diagnosis when the hepatic lesion is complicated by the presence of greatly elevated alpha fetoprotein (AFP). Clinical treatment of non-typical hepatic pseudolesions complicated by greatly elevated AFP should confirm the diagnosis and minimize trauma. CASE PRESENTATION: Non-invasive procedures including ultrasonography, CT, and micro-invasive digital subtraction angiography could not safely differentiate this lesion from a malignant focus when it was complicated by greatly elevated AFP. Laparoscopic hepatectomy was performed, and pathological analysis showed chronic hepatitis, nodular regenerative hyperplasia, focal nodular hyperplasia of the liver, and mild vascular malformation. The tissue was HbsAg(-), HbcAg(-), and AFP(+). CONCLUSION: Heightened awareness of hepatic pseudolesion complicated by primarily elevated AFP will help physicians avoid unnecessary invasive procedures. Hepatic biopsy is inevitable because of greatly elevated AFP. For suspected hepatic pseudolesion with elevated AFP, needle-core biopsy and follow-up surveillance instead of hepatectomy are recommended to find the source of AFP and make a final diagnosis of pseudolesion.
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Hepatopatías/sangre , Hepatopatías/terapia , alfa-Fetoproteínas/metabolismo , Adulto , Femenino , Hepatectomía , Hepatitis Crónica/sangre , Humanos , Hiperplasia , Laparoscopía , Hígado/irrigación sanguínea , Hígado/patología , Hepatopatías/diagnóstico , Pronóstico , Tomografía Computarizada por Rayos X , Malformaciones Vasculares/sangre , Adulto JovenRESUMEN
Objective: To investigate the expression, prognosis, and underlying mechanism of Paxillin (PXN) in ovarian cancer. Materials and methods: By comprehensive use of various bioinformatics tools, we analyzed the expression of PXN and its prognostic value in ovarian cancer. Then, the enrichment analyses were conducted to determine the possible regulatory pathways PXN involved in ovarian cancer. Finally, the associations of PXN expression with immune cell infiltration and immune checkpoints were analyzed. Results: PXN was highly expressed in ovarian cancer and its expression could independently predict the overall survival of ovarian cancer patients. More importantly, PXN had a superior ability in predicting long-term survival than age and tumor residual disease in ovarian cancer patients. In addition, PXN was positively related to adherens junction and tight junction pathways. Significant negative relationships between PXN expression and immune infiltrates were observed, however, PXN was positively connected with immune checkpoint (VSIR) in ovarian cancer. Conclusions: PXN serves as a reliable prognostic biomarker and may be a potent therapeutic target for ovarian cancer. Moreover, high PXN expression may affect ovarian cancer progression via positive regulation of metastasis-related pathways.
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OBJECTIVE: The study goal was to determine the diagnostic accuracy of a specific cytokine pattern including interferon-gamma (IFN-γ), interleukin (IL)-10, and IL-6 for hemophagocytic lymphohistiocytosis (HLH) in febrile children. STUDY DESIGN: In this prospective study, 756 patients with fever admitted to a hematology-oncology unit were enrolled. The causes of fever were documented and the serum cytokines, including IFN-γ, tumor necrosis factor-alpha (TNF-α), IL-10, IL-6, IL-4, and IL-2, were determined using cytometric bead array techniques. RESULTS: Of 1474 episodes of fever that were analyzed, 71 episodes of HLH manifested a specific cytokine pattern of highly increased levels of IFN-γ (median level: 1088.5 pg/mL) and IL-10 (623.5 pg/mL) but a moderately increased level of IL-6 (51.1 pg/mL). IL-6 was predominantly increased to varied extents in patients in the sepsis group (244.6 pg/mL) and the nonsepsis infection group (34.7 pg/mL). The diagnostic accuracy of IFN-γ and IL-10 for HLH was 99.5% and 92.8%, respectively. By applying the cutoff point of 100 pg/mL, IFN-γ had a sensitivity of 94.4% and a specificity of 97.2% for HLH. When using the criteria of IFN-γ >75 pg/mL and IL-10 >60 pg/mL, the specificity reached 98.9% and the sensitivity was 93.0%. CONCLUSIONS: The specific cytokine pattern of markedly elevated levels of IFN-γ and IL-10 with only modestly elevated IL-6 levels has high diagnostic accuracy for HLH and may be a useful approach to differentiate HLH from infection.
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Citocinas/sangre , Linfohistiocitosis Hemofagocítica/diagnóstico , Adolescente , Biomarcadores/sangre , Niño , Preescolar , Diagnóstico Diferencial , Femenino , Estudios de Seguimiento , Humanos , Lactante , Recién Nacido , Interferón gamma/sangre , Interleucina-10/sangre , Interleucina-6/sangre , Linfohistiocitosis Hemofagocítica/sangre , Masculino , Estudios Prospectivos , Reproducibilidad de los Resultados , Factor de Necrosis Tumoral alfa/sangreRESUMEN
OBJECTIVE: To search for the most suitable gene transfection conditions for rat muscle in vivo by therapertic ultrasound-mediated microbubble destruction (UMMD). METHODS: A mixture of microbubbles and enhanced green flurescence protein (EGFP) plasmids was injected into rat tibialis anterior muscle and the muscels treated with ultrasound irradiation by different output intensity, duty cycle and irradiation time of therapeutic ultrasound. The transfection efficiency was demonstrated by the EGFP expression results under fluorescent staining and immunohistochemical staining. And the most favorable ultrasound conditions as well as local intra-muscle and intravenous injection methods were selected based on the best transfection efficiency and the least muscle damage under HE staining. The rats were divided into four groups: 1) ultrasound + microbubbles + plasmid; 2) microbubble + plasmid; 3) ultrasound + plasmid; 4) plasmid only. The favorable ultrasound conditions and injection method were selected out on the basis of above steps. EGFP expression was observed in the tibialis anterior muscle of each group. The rats were sacrificed in groups at 5 days after they underwent ultrasound irradiation, the EGFP expression in muscle was observed after fluorescent and immunohistochemical stainingand the muscle damage was also observed under HE staining. RESULTS: The most favorable conditions consisted of a 1-MHz therapeutic ultrasound irradiation applied for 3 min, a power output of 2 W/cm2 and a 20% duty cycle. Under these conditions, the muscle showed significant EGFP expression, and the muscle was not significantly damaged. The EGFP expression induced by the local intra-muscle injection was more significantly increased than that induced by the intravenous injection. Among the four groups, the EGFP expression under fluorescence staining and immunohistochemical staining in the muscle of the ultrasound + microbubbles + plasmid group was significantly higher than that of the other three groups, and the microbubbles + plasmid group was higer than that of the other two groups (P<0.05). No muscle damage caused by the ultrasound and microbubbles was detected under HE staining. CONCLUSION: Under the optimal transfection conditions, the therapeutic ultrasound-mediated microbubble destruction method can significantly enhance the in vivo gene transfection efficiency of rat muscle and found no damage to the muscle. Thus, these conditions can be used as part of a safe and effective non-viral gene transfection procedure for gene therapy.
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Técnicas de Transferencia de Gen , Microburbujas , Músculo Esquelético/metabolismo , Transfección/métodos , Ultrasonido/métodos , Animales , Femenino , Terapia Genética , Proteínas Fluorescentes Verdes/metabolismo , Masculino , Ratas , Ratas WistarRESUMEN
OBJECTIVES: To evaluate the determinants of failure rate and reproducibility of computed tomography feature tracking (CT-FT) derived left ventricular (LV) strain. METHODS: Subjects who underwent retrospectively electrocardiogram gated coronary computed tomography angiography (CCTA) were included in this study. CCTA image quality and strain curve were visually evaluated to determine whether the images were optimal for CT-FT strain analysis. Parameters of global strain, regional longitudinal strain (LS), territory LS, segmental LS, and LS mechanical dispersion were obtained. Reproducibility was quantified by intraclass correlation coefficient (ICC), Bland-Altman analysis, and coefficient of variation (CV). RESULTS: A total of 809 subjects were included in the final study cohort, and 625 subjects had complete LV CT-FT strain analysis. The risk factors for failed LV strain analysis were high average heart rate (odds ratios [OR] = 5.52; 95% confidence interval [95 %CI]: 3.59-8.68), high heart rate variation (OR = 2.7; 95% CI = 1.54-3.68), and older age (OR = 1.87; 95% CI = 1.24-2.84). Tube current modulation was inversely related to failure of LV strain analysis (OR = 0.19; 95% CI = 0.11-0.34). The intraobserver reproducibility of global strain was excellent (ICCs: 0.90-0.98, CVs: 3.4-9.7%), and the interobserver reproducibility was excellent (ICCs: 0.84-0.96, CVs: 4.4-11.9%), except for global radial strain. The intra- and interobserver ICCs of territory LS ranged from 0.83 to 0.97 and 0.76 to 0.95, while the CVs ranged from 3.7% to 9.2% and 4.6% to 10.6%.The intra- and inter-observer ICCs of regional LS ranged from 0.82 to 0.95 and 0.79 to 0.94, while the CVs ranged from 5.5% to 10.2% and 5.7% to 11.0%. The reproducibility of segmental LS was variable, and the ICCs were below 0.75 in 60.8% (31/51) segments. The reproducibility of LS mechanical dispersion was poor (the ICCs were below 0.50, and the CVs were greater than 20%). CONCLUSIONS: Controlling heart rate during CT scanning is essential to perform a successful analysis of LV CT-FT strain. The reproducibility of CT-FT-derived global strain, regional LS, and territory LS was good to excellent, while the segmental LS should be used with caution in clinical practice, and LS mechanical dispersion is not recommended at present.
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Imagen por Resonancia Cinemagnética , Función Ventricular Izquierda , Humanos , Imagen por Resonancia Cinemagnética/métodos , Variaciones Dependientes del Observador , Reproducibilidad de los Resultados , Estudios Retrospectivos , Tomografía Computarizada por Rayos X , Función Ventricular Izquierda/fisiologíaRESUMEN
Several studies indicated that autophagy activation participates in brain ischemic tolerance (BIT) induced by cerebral ischemic preconditioning (CIP). However, the mechanism of autophagy activation during the process still remains unclear. The present study aimed to evaluate the role of p38 MAPK-peroxisome proliferator-activated receptor γ (PPARγ) signaling cascade in autophagy during the CIP-induced BIT. The results shown that, initially, autophagy activation was observed after CIP in the model of global cerebral ischemia in rats, as was indicated by the upregulation of Beclin 1 expression, an increase in LC3-II/LC3-I ratio, the enhanced LC3 immunofluorescence, and a rise in the number of autophagosomes in the neurons of the hippocampal CA1 area. Besides, the inhibitor of autophagy 3-methyladenine obliterated the neuroprotection induced by CIP. Furthermore, the upregulation of p-p38 MAPK and PPARγ expressions was earlier than autophagy activation after CIP. In addition, pretreatment with SB203580 (the inhibitor of p38 MAPK) reversed CIP-induced PPARγ upregulation, autophagy activation, and neuroprotection. Pretreatment with GW9662 (the inhibitor of PPARγ) reversed autophagy activation and neuroprotection, while it had no effect on p-p38 MAPK upregulation induced by CIP. These data suggested that the p38 MAPK-PPARγ signaling pathway participates in autophagy activation during the induction of BIT by CIP.
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Isquemia Encefálica , Precondicionamiento Isquémico , Animales , Autofagia , Encéfalo/metabolismo , Isquemia Encefálica/metabolismo , Precondicionamiento Isquémico/métodos , PPAR gamma/genética , PPAR gamma/metabolismo , Ratas , Ratas Wistar , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismoRESUMEN
OBJECTIVE: To optimize the conditions of ultrasonic irradiation and microbubble of ultrasound cavitation on destruction of synovial pannus of antigen-induced arthritis (AIA) in rabbits. METHODS: Antigen-induced arthritis was successfully induced on bilateral knee joints of 85 rabbits. Each 10 AIA rabbits were divided into two groups to compare various peak negative pressures, different ultrasonic pulse durations, various pulse repetition frequencies, different irradiance duration, different dosages of microbubble contrast agents, different ultrasonic irradiance times. With intravenous infusion of Sonovue to the rabbits, ultrasonic irradiance was performed on the right knee joint using the above condition of ultrasound cavitation. At the day 1 after ultrasonic irradiance, MRI and pathological examination were employed to evaluate the optimal conditions. RESULTS: The optimal parameters and conditions for ultrasonic irradiance included intermittent ultrasonic application (in 6 s intervals), 0.6 mL/kg of microbubble contrast agent, 4.6 MPa of ultrasonic peak negative pressure, 100 cycles of pulse duration, 50 Hz of pulse repetition frequency, 5 min of ultrasonic duration, 0.6 mL/kg of dosages of microbubble contrast agents and multi-sessional ultrasonic irradiance. After the ultrasonic irradiance, the thickness of right knee synovium measured by MRI was thinner than that of left knee and synovial necrosis was confirmed by the pathological finding. CONCLUSION: Under optimal ultrasonic irradiation and microbubble conditions, ultrasonic cavitation could destroy synovial pannus of AIA in rabbits.
Asunto(s)
Artritis Experimental/terapia , Microburbujas/uso terapéutico , Sonido , Membrana Sinovial/irrigación sanguínea , Terapia por Ultrasonido/métodos , Animales , Artritis Experimental/inducido químicamente , Artritis Experimental/patología , Femenino , Masculino , Ovalbúmina , Conejos , Membrana Sinovial/efectos de la radiaciónRESUMEN
Carbon black (CB) has been demonstrated to have adverse effects on the lung tissue. Few studies explored the effects of CB on the cerebellum, widely recognized to contribute to gait and balance coordination and timing in the motor domain. Some studies have reported that inflammatory response and damaged autophagy are important mechanisms of CB toxicity and can be repaired after the recovery. The present study aimed to determine whether long-term CB exposure could induce the inflammation and damaged autophagy of the cerebellum. The rats were randomly divided into four groups. The control group received the filtered air for 90 days; the carbon black (CB) group received CB particles for 90 days; the recovery (R) group received CB for 90 days and recovered for another 14 days; the recovery control (RC) group received filtered air for 104 days. The purpose of the R group was to test whether neuroinflammation and autophagy could be repaired after short-term recovery. The western blot and immunohistochemistry revealed that long-term CB exposure induced augmented level of pro-inflammatory cytokines (Interleukin-1ß, IL-1ß; Interleukin-6, IL-6; and Tumor Necrosis Factor-α, TNF-α) and anti-inflammatory cytokine (Interleukin-10, IL-10). The autophagic markers (Beclin1 and LC3) were increased in both CB group and R group. These findings clearly demonstrated that long-term CB exposure induced inflammation and autophagy in the cerebellum, which were not obviously improved after short-term recovery.
Asunto(s)
Autofagia/efectos de los fármacos , Cerebelo/efectos de los fármacos , Enfermedades Neuroinflamatorias/inducido químicamente , Hollín/toxicidad , Animales , Western Blotting , Cerebelo/patología , Masculino , Enfermedades Neuroinflamatorias/patología , Ratas , Ratas Sprague-Dawley , Hollín/administración & dosificaciónRESUMEN
Our previous finding suggests that p38 MAPK contributes to the GLT-1 upregulation during induction of brain ischemic tolerance by cerebral ischemic preconditioning (CIP), however, the underlying mechanism is still unclear. Here, we investigated the molecular mechanisms underlying the CIP-induced GLT-1 upregulation by using Western blotting, Co-immunoprecipitation (Co-IP), electrophoretic mobility shift assay (EMSA) and thionin staining in rat hippocampus CA1 subset. We found that application of BAY11-7082 (an inhibitor of NF-κB), or dihydrokainate (an inhibitor of GLT-1), or SB203580 (an inhibitor of p38 MAPK) could attenuate the CIP-induced neuronal protection in hippocampus CA1 region of rats. Moreover, CIP caused rapid activation of NF-κB, as evidenced by nuclear translocation of NF-κB p50 protein, which led to active p50/p65 dimer formation and increased DNA binding activity. GLT-1 was also increased after CIP. Pretreatment with BAY11-7082 blocked the CIP-induced GLT-1 upregulation. The above results suggest that NF-κB participates in GLT-1 up-regulation during the induction of brain ischemic tolerance by CIP. We also found that pretreatment with SB203580 caused significant reduction of NF-κB p50 protein in nucleus, NF-κB p50/p65 dimer nuclear translocation and DNA binding activity of NF-κB. Together, we conclude that p38 MAPK/NF-κB pathway participates in the mediation of GLT-1 up-regulation during the induction of brain ischemic tolerance induced by CIP.