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Phylogenetically and antigenically distinct influenza A and B viruses (IAV and IBV) circulate in human populations, causing widespread morbidity. Antibodies (Abs) that bind epitopes conserved in both IAV and IBV hemagglutinins (HAs) could protect against disease by diverse virus subtypes. Only one reported HA Ab, isolated from a combinatorial display library, protects against both IAV and IBV. Thus, there has been so far no information on the likelihood of finding naturally occurring human Abs that bind HAs of diverse IAV subtypes and IBV lineages. We have now recovered from several unrelated human donors five clonal Abs that bind a conserved epitope preferentially exposed in the postfusion conformation of IAV and IVB HA2. These Abs lack neutralizing activity in vitro but in mice provide strong, IgG subtype-dependent protection against lethal IAV and IBV infections. Strategies to elicit similar Abs routinely might contribute to more effective influenza vaccines.
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Vírus da Influenza A , Vacinas contra Influenza , Influenza Humana , Infecções por Orthomyxoviridae , Humanos , Camundongos , Animais , Hemaglutininas , Epitopos , Anticorpos Antivirais , Glicoproteínas de Hemaglutininação de Vírus da Influenza , Vírus da Influenza BRESUMO
Background: Recent studies have demonstrated that activated microglia were involved in the pathogenesis of central sensitization characterized by cutaneous allodynia in migraine. Activation of microglia is accompanied by increased expression of its receptors and release of inflammatory mediators. Acupuncture and its developed electroacupuncture (EA) have been recommended as an alternative therapy for migraine and are widely used for relieving migraine-associated pain. However, it remains rare studies that show whether EA exerts anti-migraine effects via inhibiting microglial activation related to a release of microglial receptors and the inflammatory pathway. Therefore, this study aimed to investigate EA' ability to ameliorate central sensitization via modulation of microglial activation, microglial receptor, and inflammatory response using a rat model of migraine induced by repeated epidural chemical stimulation. Methods: In the present study, a rat model of migraine was established by epidural repeated inflammatory soup (IS) stimulation and treated with EA at Fengchi (GB20) and Yanglingquan (GB34) and acupuncture at sham-acupoints. Pain hypersensitivity was further determined by measuring the mechanical withdrawal threshold using the von-Frey filament. The changes in c-Fos and ionized calcium binding adaptor molecule 1 (Ibal-1) labeled microglia in the trigeminal nucleus caudalis (TNC) were examined by immunflurescence to assess the central sensitization and whether accompanied with microglia activation. In addition, the expression of Ibal-1, microglial purinoceptor P2X4, and its associated inflammatory signaling pathway mediators, including interleukin (IL)-1ß, NOD-like receptor protein 3 (NLRP3), and Caspase-1 in the TNC were investigated by western blot and real-time polymerase chain reaction analysis. Results: Allodynia increased of c-Fos, and activated microglia were observed after repeated IS stimulation. EA alleviated the decrease in mechanical withdrawal thresholds, reduced the activation of c-Fos and microglia labeled with Ibal-1, downregulated the level of microglial purinoceptor P2X4, and limited the inflammatory response (NLRP3/Caspase-1/IL-1ß signaling pathway) in the TNC of migraine rat model. Conclusions: Our results indicate that the anti-hyperalgesia effects of EA ameliorate central sensitization in IS-induced migraine by regulating microglial activation related to P2X4R and NLRP3/IL-1ß inflammatory pathway.
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Modelos Animais de Doenças , Eletroacupuntura , Hiperalgesia , Inflamação , Microglia , Transtornos de Enxaqueca , Ratos Sprague-Dawley , Receptores Purinérgicos P2X4 , Animais , Eletroacupuntura/métodos , Receptores Purinérgicos P2X4/metabolismo , Microglia/metabolismo , Hiperalgesia/terapia , Hiperalgesia/metabolismo , Transtornos de Enxaqueca/terapia , Transtornos de Enxaqueca/metabolismo , Masculino , Inflamação/metabolismo , Inflamação/patologia , Inflamação/terapia , Sensibilização do Sistema Nervoso Central/fisiologia , Ratos , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Proteínas Proto-Oncogênicas c-fos/metabolismoRESUMO
Background: The nursing work in the operating room is heavy, intensive, and irregular, and the quality of nursing work can directly affect the surgical effect and patient prognosis. Therefore, nursing management in the operating room should be strengthened to protect patients' life safety effectively. Objective: To assess the effectiveness of applying the 6S management model in operating room nursing. Design: This was a retrospective study. Setting: This study was conducted at the Department of Anesthesia Surgery, Nanfang Hospital, Southern Medical University. Participants: The research included 100 operating room nurses on duty between January 2020 and December 2022. Intervention: From January 2020 to June 2021, the hospital conducted routine training programs for nurses in the operating room. From July 2021 to December 2022, the hospital adopted the 6S management model for overseeing nursing work in the operating room. Primary Outcome Measures: (1) nursing quality score (2) nursing staff safety awareness (3) nursing disputes and complaints (4) incidence of adverse reactions (5) patient satisfaction with the quality of nursing care. Results: Following the adoption of the 6S management model, there was a noticeable improvement in the nursing quality scores, the nursing staff's awareness of safety, and the satisfaction levels of patients with the quality of care provided by operation nurses (P < .05). Additionally, the incidence of nursing disputes, complaints, and adverse events among patients decreased significantly compared to before the implementation of 6S (P = .01). Conclusion: Implementing 6S management with a focus on the work of operation room nurses enhanced the competence of the nursing staff and improved management effectiveness, ultimately leading to increased patient satisfaction.
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To overcome the limitations of NO2 electrochemical sensors, including their inaccurate measurements and short working life, when used around coal-fired power plants, we investigated the influence of coal-fired fly ash deposition on the measurement error of NO2 electrochemical sensors through experimental tests. The morphological characteristics and pellet diameter distribution of coal-fired fly ash pellets were determined via scanning electron microscopy. The sedimentation velocity of coal-fired fly ash pellets in the air was determined through theoretical calculations of aerodynamics and hydrodynamics. Additionally, the effect of the deposition of coal-fired fly ash on the measurement error of NO2 electrochemical sensors was determined through experimental tests. The test results show that the minimum and maximum measurement errors of the NO2 electrochemical gas sensor were 8.015% and 30.35%, respectively, after a deposition duration of 30 days with 30 mg/m3 coal-fired fly ash. This demonstrates that coal-fired fly ash deposition is the cause of the inaccurate measurements and short working life of these sensors. Coal-fired fly ash causes a decrease in the gas diffusion area of the sensor and the diffusion coefficient, thus increasing the sensor measurement error.
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BACKGROUND: Renal fibrosis is a final common pathway of chronic kidney disease. SIRT1, a NAD+-dependent protein deacetylase, deacetylates the p65 of NF-κB and shows protective effects in kidney disorders. miR-373 directly targets the 3'UTR of SIRT1. However, roles of miR-373 in renal fibrosis are unclear. METHODS: TGF-ß1, a critical regulator of fibrosis, was used to stimulate human kidney-2 cells to establish cell model for renal fibrosis. Unilateral ureteral obstruction (UUO) was performed as an in vivo model. RESULTS: TGF-ß1 induced the level of miR-373, reduced level of SIRT1, and promoted p65 acetylation and MMP-9 expression. These effects were reversed by the miR-373 inhibitor. In the animal model, UUO caused a consistent pattern as demonstrated in vitro. CONCLUSION: These results indicated an undesired effect of miR-373 in the regulation of renal injury and fibrosis by targeting SIRT1-mediated NF-κB/MMP-9 signaling, which might provide a potential therapeutic strategy for renal fibrosis.
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Rim/metabolismo , MicroRNAs/fisiologia , Insuficiência Renal/metabolismo , Sirtuína 1/metabolismo , Animais , Linhagem Celular , Fibrose , Humanos , Rim/patologia , Metaloproteinase 9 da Matriz/genética , Metaloproteinase 9 da Matriz/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , MicroRNAs/genética , NF-kappa B/genética , NF-kappa B/metabolismo , Insuficiência Renal/etiologia , Transdução de Sinais , Sirtuína 1/genética , Obstrução Ureteral/complicaçõesRESUMO
The pioneering prediction and successful synthesis of monolayer arsenene in recent years have promoted intensive studies on this novel two-dimensional (2D) material. Strain-engineered arsenene monolayer can change its geometric structures with tuned charge distribution, which paves the way for achieving novel electronic properties. The practical applications of the strain-driven topological state in arsenene strongly depend on its critical strain value. In this work, mechanical properties such as fracture strain, fracture strength and Young's modulus of two arsenene structures, i.e. buckled arsenene (b-arsenene) and puckered arsenene (p-arsenene), are comprehensively investigated under different modulators such as system dimension, chirality, temperature, strain rate and random surface defect. A maximum fracture strain reduction of 41.7% from 0.24 to 0.14 is observed in armchair b-arsenene when the temperature increases from 100 to 500 K. The most significant impact factor on the mechanical properties of arseneneis found to be surface defects. A maximum fracture strength reduction of 85.7% is predicted in the armchair b-arsenene when the defect ratio increases from 0 to 5%. On the other hand, the strain rate has a negligible effect on the mechanical properties. Our results provide fundamental knowledge on the critical fracture properties of arsenene.
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The Aicda gene product, activation-induced cytidine deaminase (AID), initiates somatic hypermutation, class-switch recombination, and gene conversion of Ig genes by the deamination of deoxycytidine, followed by error-prone mismatch- or base-excision DNA repair. These processes are crucial for the generation of genetically diverse, high affinity antibody and robust humoral immunity, but exact significant genetic damage and promote cell death. In mice, physiologically significant AID expression was thought to be restricted to antigen-activated, mature B cells in germinal centers. We now demonstrate that low levels of AID in bone marrow immature and transitional B cells suppress the development of autoreactivity. Aicda(-/-) mice exhibit significantly increased serum autoantibody and reduced capacity to purge autoreactive immature and transitional B cells. In vitro, AID deficient immature/transitional B cells are significantly more resistant to anti-IgM-induced apoptosis than their normal counterparts. Thus, early AID expression plays a fundamental and unanticipated role in purging self-reactive immature and transitional B cells during their maturation in the bone marrow.
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Linfócitos B/enzimologia , Linfócitos B/imunologia , Citidina Desaminase/imunologia , Tolerância a Antígenos Próprios/imunologia , Animais , Apoptose , Autoanticorpos/sangue , Linfócitos B/citologia , Ligante de CD40/deficiência , Ligante de CD40/genética , Ligante de CD40/imunologia , Diferenciação Celular , Citidina Desaminase/deficiência , Citidina Desaminase/genética , Feminino , Genes de Cadeia Pesada de Imunoglobulina , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Células Precursoras de Linfócitos B/citologia , Células Precursoras de Linfócitos B/enzimologia , Células Precursoras de Linfócitos B/imunologia , Receptores de Antígenos de Linfócitos B/metabolismo , Tolerância a Antígenos Próprios/genética , Transdução de Sinais/imunologia , Hipermutação Somática de ImunoglobulinaRESUMO
BACKGROUND: Multiple studies have suggested that paralgesia (hyperalgesia and cutaneous allodynia) in migraine reflects the activation and sensitisation of the trigeminovascular system (TGVS). In particular, it reflects the second-order and higher nerve centre sensitisation, which is caused and maintained by neuroinflammation. Microglia activation leads to the release of proinflammatory cytokines involved in inflammatory responses. Accumulating evidence indicates that electroacupuncture (EA) is effective in ameliorating paralgesia, but the underlying mechanisms of EA in migraine attacks caused by microglia and microglia-mediated inflammatory responses are still unclear. The purpose of this study was to explore whether EA could ameliorate the dysregulation of pain sensation by suppressing microglial activation and the resulting neuroinflammatory response, and to evaluate whether this response was regulated by Toll-like receptor 4 (TLR4)/nuclear factor-kappa B(NF-κB) in the trigeminal nucleus caudalis (TNC) in a rat model of migraine. METHODS: Repeated Inflammatory Soup (IS) was infused into the dura for seven sessions to establish a recurrent migraine-like rat model, and EA treatment was administered at Fengchi (GB20) and Yanglingquan (GB34) after daily IS infusion. Facial mechanical withdrawal thresholds were measured to evaluate the change in pain perception, and plasma samples and the TNC tissues of rats were collected to examine the changes in calcitonin gene-related peptide (CGRP), the Ibal-1-labelled microglial activation, and the resulting inflammatory response, including interleukin-1ß (IL-1ß), tumour necrosis factor-α (TNF-α), interleukin-6 (IL-6), and their regulatory molecules TLR4/NF-κB, via enzyme-linked immunosorbent assay (ELISA), real-time polymerase chain reaction (RT-PCR), immunohistochemistry (IHC) and Western blot analysis. RESULTS: Repeated IS injections into the dura induced facial mechanical paralgesia, which is the manifestation of migraine attacks, and increased the expression of CGRP, Ibal-1, microglial mediated inflammatory cytokines (IL-1ß, TNF-α, IL-6), and regulatory molecules TLR4/NF-κB. EA at GB20/34 significantly attenuated repetitive IS-induced pain hypersensitivity. This effect was consistent with decreased levels of CGRP and inflammatory cytokines in the plasma and the TNC via the inhibition of microglia activation, and this response may be regulated by TLR4/NF-κB. CONCLUSIONS: EA ameliorated paralgesia in repetitive IS-induced migraine-like rats, which was mainly mediated by a reduction in microglial activation and microglial-mediated inflammatory responses that could be regulated by TLR4/NF-κB.
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Depression is a complex clinical disorder associated with poor outcomes. Electroacupuncture (EA) has been demonstrated to have an important role in both clinical and pre-clinical depression investigations. Evidence has suggested that the P2X7 receptor (P2X7R), NLRP3, and IL-1ß play an important role in depressive disorder. Our study is aimed at exploring the role of EA in alleviating depression-like behaviors in rats. We therefore investigated the effects of EA on the prefrontal cortex and liver of rats subjected to chronic unpredictable mild stress (CUMS) through behavior tests, transmission electron microscopy, Nissl staining, HE staining, immunohistochemistry and Western blotting. Five weeks after exposure to CUMS, Sprague-Dawley (SD) rats showed depression-like behavior. Three weeks after treatment with brilliant blue G (BBG) or EA, depressive symptoms were significantly improved. Liver cells and microglia showed regular morphology and orderly arrangement in the BBG and EA groups compared with the CUMS group. Here we show that EA downregulated P2X7R/NLRP3/IL-1ß expression and relieved depression-like behavior. In summary, our findings demonstrated the efficacy of EA in alleviating depression-like behaviors induced by CUMS in rats. This suggests that EA may serve as an adjunctive therapy in clinical practice, and that P2X7R may be a promising target for EA intervention on the liver-brain axis in treatment of depression.
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Glioma is one of the most frequent types of primary tumors in central nervous system. Previous studies deomostrated that tumor-associated macrophages (TAMs) and their marker genes were significantly associated with immunologic suppression and immune escape of cancer. However, the molecular mechanism between glioma and TAM marker genes is still rarely reported. In this research, we performed a comprehensive analysis of the prognostic prediction value of TAM marker genes in multiple glioma cohorts. Further investigation indicated that the increased expression of TAM marker genes resulted in the immune suppressive microenvironment in glioma through regulating tumor-infiltrating immune cells and Cancer-Immunity Cycle. To better forecast the survival of glioma patients, we then developed gene risk models in four glioma datasets (CGGA, TCGA, Rembrandt and Gravendeel). Univariate and multivariate Cox analysis exhibited the good survival prediction ability and prognostic discrimination ability of our models. The results of immunotherapy prediction indicated that glioma patients with low risk were more likely to benefit from ICB (immune checkpoint blockade) treatment. Altogether, our research provided a comprehensive analysis of TAM marker genes and explored their value for predicting prognosis and immunotherapy response in glioma.
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Neoplasias Encefálicas , Glioma , Biomarcadores Tumorais/genética , Neoplasias Encefálicas/genética , Neoplasias Encefálicas/patologia , Neoplasias Encefálicas/terapia , Perfilação da Expressão Gênica , Glioma/genética , Glioma/patologia , Glioma/terapia , Humanos , Imunoterapia , Microambiente Tumoral/genética , Macrófagos Associados a TumorRESUMO
Many of the best HIV-1 broadly neutralizing antibodies (bnAbs) known have poly-/autoreactive features that disfavor normal B cell development and maturation, posing a major hurdle in developing an effective HIV-1 vaccine. Key to resolving this problem is to understand if, and to what extent, neutralization breadth-conferring mutations acquired by bnAbs contribute to their autoreactivity. Here, we back-mutated all known changes made by a prototype CD4 binding site-directed bnAb lineage, CH103-106, during its later maturation steps. Strikingly, of 29 mutations examined, only four were crucial for increased autoreactivity, with minimal or no impact on neutralization. Furthermore, three of these residues were clustered in the heavy chain complementarity-determining region 2 (HCDR2). Our results demonstrate that broad neutralization activity and autoreactivity in the CH103-106 bnAb lineage can be governed by a few, distinct mutations during maturation. This provides strong rationale for developing immunogens that favor bnAb lineages bearing "neutralization-only" mutations into current HIV-1 vaccine designs.
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HIV-1 , Vacinas , HIV-1/genéticaRESUMO
OBJECTIVE: To observe the effect of mild moxibustion on monocyte chemotaxis protein 1 (MCP-1)/matrix metalloproteinase 2 (MMP-2)/transforming growth factor ß1 (TGF-ß1) pro-inflammatory signal loop in senile rats, so as to explore its mechanisms underlying improving vascular aging (VA). METHODS: Twenty-four male VA SD rats were randomized into senium (VA) control, medication and moxibustion groups, and other 8 young SD rats (aged 2 months) were used as the young control group. The VA model was established by intraperitoneal injection of D-galactose (300 mg·kg-1·d-1) once daily for 4 weeks, and verified by serum total testosterone (TT) and free testosterone (FT) levels. For rats of the moxibustion group, mild moxibustion was applied to bilateral "Shenshu"(BL23) and "Guanyuan"(CV4) for 20 min, once a day, 5 days a week for 8 weeks. Rats of the medication group were treated by intraperitoneal injection of testosterone propionate (7 mg·kg-1·[3 d]-1) once daily for 8 weeks except weekends, and rats of the senium control and young control groups treated by intraperitoneal injection of the same dose of normal saline, once daily for 8 weeks except weekends. The duration of exhausted swimming (DES) before and after the treatment was recorded. H.E. staining and Masson staining were used to observe histopathological changes and collagen fiber content of the thoracic aortic tissue, respectively. The contents of serum TT, FT and angiotensin 2 (Ang â ¡) were determined by ELISA. The immunoactivity of aortic MCP-1 was detected by immunohistochemistry, and the expression levels of aortic MCP-1, MMP-2 and TGF-ß1 were detected by Western blot. RESULTS: Compared with the young control group, the levels of DES, serum TT and FT contents were significantly decreased (P<0.01), while those of serum Angâ ¡ and collagen fiber contents, aortic MCP-1 immunoactivity and MMP-2, TGF-ß1 and MCP-1 protein expression considerably increased in the senium control group (P<0.01). After the interventions, the decreased levels of DES, serum TT and FT contents and the increased levels of serum Angâ ¡, collagen fiber contents, aortic MCP-1 immunoactivity and MMP-2, TGF-ß1 and MCP-1 protein expression were reversed in both medication and moxibustion groups (P<0.01, P<0.05). The therapeutic effect of mild moxibustion was significantly superior to that of medication in down-regulating the aortic collagen fiber, serum Angâ ¡ contents and MCP-1 immunoactivity and protein expression (P<0.05). H.E. staining showed thickened endometrium and disordered arrangement of vascular smooth muscles of the aorta in the senium group, and thinner endometrium and regular and ordered arrangement of aortic vascular smooth muscles in both moxibustion and medication groups. CONCLUSION: Mild moxibustion may improve vascular aging in senescence rats, which is possibly by suppressing vascular MCP-1/MMP-2/TGF-ß1 pro-inflammatory signal loop.
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Moxibustão , Envelhecimento , Animais , Colágeno , Feminino , Masculino , Metaloproteinase 2 da Matriz/genética , Ratos , Ratos Sprague-Dawley , Testosterona , Fator de Crescimento Transformador beta1/genéticaRESUMO
OBJECTIVE: To observe the effect of electroacupuncture (EA) on the expressions of tight junction related proteins Claudin-5, ZO-1 in the colon and hippocampus, Toll-like receptor 4/nuclear factor-kappa B/NOD-like receptor protein 3 (TLR4/NF-κB/NLRP3) pathway in the hippocampus of APP/PS1 mice, so as to explore its mechanisms underlying improvement of cognitive impairment. METHODS: Eighteen 5-month-old male APP/PS1 mice were equally randomized into model and EA groups,and nine 5-month-old male C57BL/6 mice were used as the normal control. EA(2 Hz, 1 mA) was applied to "Baihui" (GV20), "Dachangshu" (BL25) and "Zusanli" (ST36) for 15 min, once daily, 5 days a week for 5 weeks. The Morris water maze swimming test was used to evaluate the mice's cognitive impairment. Nissl staining was used to observe the pathological morphology of hippocampus. The expression of amyloid ß-peptide (Aß) in brain tissue was detect by immunohistochemistry; the contents of lipopolysaccharide (LPS) in colon, serum and hippocampus were detected by ELISA; the expression levels of Claudin-5, ZO-1 in colon and hippocampus, and TLR4/NF-κB/NLRP3 pathway related proteins in hippocampus were detected by Western blot. RESULTS: Compared with the normal group, the escape latency of the mice in the model group was prolonged from the 3rd day (P<0.05, P<0.01), the number of crossing the platform and the percentage of target quadrant residence time were significantly decreased (P<0.01), and the contents of LPS in colon, serum and hippocampus were significantly increased (P<0.01), the expression levels of TLR4, NF-κB p65, NLRP3, Caspase-1, interleukin (IL)-1ß and tumor necrosis factor (TNF)-α in hippocampus and Aß in brain tissue were significantly increased (P<0.01), while the expression levels of Claudin-5, ZO-1 in colon and hippocampus were significantly decreased (P<0.01). Compared with the model group, the escape latency of mice in the EA group was shortened from the 4th day (P<0.05, P<0.01), the number of crossing the platform and the percentage of target quadrant residence time were increased (P<0.01, P<0.05), and the contents of LPS in serum and hippocampus were decreased (P<0.05), and the expression levels of TLR4, NF-κB p65, Caspase-1, NLRP3, IL-1ß, TNF-α in hippocampus and Aß in brain tissue were significantly decreased (P<0.05, P<0.01), while the expression levels of Claudin-5, ZO-1 in colon and hippocampus were significantly increased (P<0.05, P<0.01). Outcomes of Nissl staining showed dispersed arrangement of neurons with nuclear pyknosis or hyperchromasia in the hippocampus, and a decreased number of cell layers in the model group, which was relatively milder in the EA group. CONCLUSION: EA may improve the cognitive impairment of APP/PS1 mice by up-regulating the expression of Claudin-5 and ZO-1, reducing the transposition of gut-derived LPS to the central nervous system, inhibiting the over-activation of TLR4/NF-κB/NLRP3 pathway, and alleviating the inflammatory reaction of the central nervous system.
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Doença de Alzheimer , Disfunção Cognitiva , Eletroacupuntura , Doença de Alzheimer/genética , Doença de Alzheimer/terapia , Peptídeos beta-Amiloides , Animais , Caspases , Claudina-5 , Disfunção Cognitiva/genética , Disfunção Cognitiva/terapia , Hipocampo , Lipopolissacarídeos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , NF-kappa B/genética , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética , Receptor 4 Toll-Like/genéticaRESUMO
OBJECTIVE: To observe the effect of electroacupuncture (EA) on physical strength and expression levels of hepatic AMP-activated protein kinase (AMPK), mammalian target of rapamycin (mTOR), unc-51 like autophagy activating kinase 1 (ULK1) proteins and Atg5, Atg7, Atg13, Beclin1 and ULK1 mRNAs in aging (senescence accelerated mouse/prone 8, SAMP8)mice, so as to exp lore its mechanism underlying delaying aging by activating AMPK/mTOR/ULK1 signaling pathway. METHODS: Twenty-four male SAMP8 mice were randomly divided into model group, rapamycin (autophagy inducer) group, EA group and EA+autophagy inhibitor (EA+inhibitor) group, with 6 mice in each group, and 6 homologous anti-rapid aging male (SAMR1) mice in the same age were used as the control group. Mice of the rapamycin group received intraperitoneal injection of rapamycin solution (2 mg·kg-1·d-1). EA (2 Hz, 1 mA) was applied to bilateral "Taichong"(LR3)and "Shenshu"(BL23) for 15 min each time. Mice of the EA+inhibitor group received intraperitoneal injection of mTOR inhibitor 3-methyladenine (1.5 mg·kg-1·d-1) before the EA intervention each time. The above-mentioned interventions were conducted 6 times a week for 2 consecutive weeks. Physical conditions of mice were assessed by exhaustive swimming tests. Histopathological changes of the liver were observed by H.E. staining. Western blot was used to detect the expression of AMPK, phosphorylated AMPK (p-AMPK), mTOR, phosphorylated mTOR (p-mTOR), ULK1 and phosphorylated ULK1 (p-ULk1) in the liver tissues. The expression levels of Atg5, Atg7, Atg13, Beclin1 and ULK1 (cellular autophagy-related genes) mRNAs in the liver were detected by quantitative real-time PCR. The immunoactivity (IA) of heme oxygenase 1 (HO-1) in the liver was detected by immunohistochemistry, and the activity of superoxide dismutase (SOD) and the content of malondialdehyde (MDA) of the liver were measured by hydroxylamine method for assessing the level of oxidative stress. RESULTS: Compared with the control group, the duration of exhaustive swimming, the expression levels of AMPK, p-AMPK, ULK1 and p-ULK1 proteins, and Atg5, Atg7, Atg13, Beclin1 and ULK1 mRNA, HO-1 IA and SOD activity were considerably down-regulated (P<0.01), while the expression levels of mTOR and p-mTOR and MDA content were significantly up-regulated (P<0.01) in the model group. In comparison with the model group, the duration of the exhausted swimming, the expression levels of AMPK, p-AMPK, ULK1 and p-ULK1 proteins, and Atg5, Atg7, Atg13, Beclin1 and ULK1 mRNAs, HO-1 IA and SOD activity were significantly up-regulated (P<0.01, P<0.05), whereas the expression levels of mTOR and p-mTOR proteins and MDA content were notably down-regulated (P<0.01, P<0.05) in the rapamycin, EA and EA+inhibitor groups. The improvement of the abovementioned indexes of EA+inhibitor group was not as good as rapamycin and EA groups (P<0.01), suggesting an elimination of the therapeutic effects after administration of 3-methyladenine. No significant differences were found between the rapamycin and EA groups in the abovementioned indexes (P>0.05) except p-mTOR and mTOR which were higher in the EA group (P<0.01). H.E. staining showed ambiguous boundary of the liver lobule, disordered arrangement of hepatocytes with a large amount of fat vacuoles at different size and deviation of nucleus, and lysis of some hepatocytes. These situations were relatively milder in the rapamycin and EA groups. CONCLUSION: EA may enhance physical strength and promote cellular autophagy in the liver of aging mice by regulating AMPK/mTOR/ULK1 signaling, thereby inhibiting excessive oxidative stress, and delaying aging process to some extent.
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Eletroacupuntura , Proteínas Quinases Ativadas por AMP/genética , Animais , Autofagia/genética , Proteína Homóloga à Proteína-1 Relacionada à Autofagia/genética , Hepatócitos , Masculino , Camundongos , Estresse Oxidativo , Ratos , Ratos Sprague-Dawley , Transdução de Sinais , Serina-Treonina Quinases TOR/genéticaRESUMO
Somatic hypermutation normally occurs as a consequence of the expression of activation-induced cytidine deaminase (AID) by Ag-activated, mature B cells during T cell-dependent germinal center responses. Nonetheless, despite their inability to express CD154 and initiate GC responses, patients with type 1 hyper-IgM syndrome (HIGM1) support populations of IgM(+)IgD(+)CD27(+) B cells that express mutated Ig genes. The origin of these mutated B cells is unknown; the IgM(+)IgD(+)CD27(+) cells do not express AID and appear to acquire mutations independent of stringent selection by Ag. Here, we demonstrate that immature/transitional 1 B cells from the bone marrow of CD154-deficient mice express AID and acquire Ig mutations that lack the hallmarks of antigenic selection via BCR signaling. Comparable levels of AID expression was found in developmentally immature B cells recovered from murine fetal liver and from human immature/transitional 1 B cells recovered from umbilical cord blood. AID expression in human fetal liver was also robust, approaching that of human tonsil tissue and the human germinal center B cell line, Ramos. These observations led us to conclude that AID expression in developing human B cells is the origin of the mutated IgM(+)IgD(+)CD27(+) B cells present in HIGM1 patients, and we propose that both mice and humans share a latent, AID-dependent pathway for the preimmune diversification of B lymphocytes that is more prominent in chicken, sheep, and rabbits.
Assuntos
Citidina Desaminase/genética , Citidina Desaminase/metabolismo , Centro Germinativo/imunologia , Centro Germinativo/patologia , Síndrome de Imunodeficiência com Hiper-IgM/enzimologia , Síndrome de Imunodeficiência com Hiper-IgM/imunologia , Animais , Subpopulações de Linfócitos B/enzimologia , Subpopulações de Linfócitos B/imunologia , Subpopulações de Linfócitos B/patologia , Células da Medula Óssea/enzimologia , Células da Medula Óssea/imunologia , Células da Medula Óssea/patologia , Linhagem Celular Transformada , Linhagem Celular Tumoral , Citidina Desaminase/biossíntese , Feminino , Regulação da Expressão Gênica no Desenvolvimento/imunologia , Rearranjo Gênico do Linfócito B/genética , Centro Germinativo/enzimologia , Humanos , Síndrome de Imunodeficiência com Hiper-IgM/genética , Imunofenotipagem , Ativação Linfocitária/genética , Ativação Linfocitária/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Dados de Sequência Molecular , Receptores de Antígenos de Linfócitos B/deficiência , Receptores de Antígenos de Linfócitos B/genética , Transdução de Sinais/genética , Transdução de Sinais/imunologia , Hipermutação Somática de Imunoglobulina/genética , Células-Tronco/enzimologia , Células-Tronco/imunologia , Células-Tronco/patologiaRESUMO
Oil granuloma (OG) induced by intraperitoneal injection of pristane represents a non-infectious granuloma. Oil granuloma has been characterized, but the regulation of its formation still remains unknown. To address this, we injected pristane into various mice deficient for genes including, linker for activation of T cells (LAT), µMT, LTα, TNFα, IL-6. T cell deficient mice (LAT(-/-) ) responded to pristane by developing serosal granuloma and mesenteric granuloma (MG) as in wild type mice. The absence of B cells blocked serosal granuloma (SG) formation and diminished MG development in response to pristane. However, even when a comparable number of B cells were present in the mesentery, the absence of TNFα resulted in similar defects in OG formation after pristane treatment, demonstrating that both B cells and TNFα are very crucial for pristane-induced OG formation. Interestingly, IL-6(-/-) mice had intact MG formation; however, SG organization was impaired. These studies provide insight into granulomateous pathology induced by non-infectious substances for example, biomedical sutures.
Assuntos
Proteínas Adaptadoras de Transdução de Sinal/genética , Granuloma , Interleucina-6/genética , Linfotoxina-alfa/genética , Proteínas de Membrana/genética , Fosfoproteínas/genética , Terpenos/toxicidade , Fator de Necrose Tumoral alfa/genética , Proteínas Adaptadoras de Transdução de Sinal/imunologia , Animais , Linfócitos B/imunologia , Feminino , Regulação da Expressão Gênica/imunologia , Granuloma/induzido quimicamente , Granuloma/genética , Granuloma/imunologia , Imunossupressores/toxicidade , Interleucina-6/imunologia , Tecido Linfoide/imunologia , Linfotoxina-alfa/imunologia , Masculino , Proteínas de Membrana/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Mutantes , Fosfoproteínas/imunologia , Fator de Necrose Tumoral alfa/imunologiaRESUMO
Granuloma formation is an inflammatory response of the host against invading pathogens or indigestible substances. We generated mesenteric oil granulomas by injecting pristane into the peritoneal cavity (PC) of mice, and compared oil granuloma formation in the C57BL/6J and BALB/cByJ strains of mice. The formation and kinetics of oil granulomas were distinct between the two strains. In C57BL/6J mice, injected pristane induced oil granuloma formation at both the mesenteric centers (MG) and margins (SG). MG was resolving by 11 weeks, and SG persisted. In BALB/cByJ mice, MG developed slower but persisted longer than in C57BL/6J mice, and SG resolved sooner than in C57BL/6J mice. Injection of India ink revealed that phagocytes were localised mainly to the SG in C57BL/6J mice, but were located diffusely in both MG and SG of BALB/cByJ mice. SG cells expressed more monocyte chemotactic protein-1 (MCP-1) mRNA than MG cells in C57BL/6J mice, but there was no difference in MCP-1 expression between the MG and SG in BALB/cByJ mice. These observations suggest that the recruitment of inflammatory leucocytes under the direction of chemokines differentiates the patterns of granuloma responses to pristane in C57BL/6J and BALB/cByJ mice.
Assuntos
Granuloma , Imunossupressores/toxicidade , Camundongos Endogâmicos BALB C/imunologia , Camundongos Endogâmicos C57BL/imunologia , Terpenos/toxicidade , Animais , Linfócitos B/imunologia , Linfócitos B/patologia , Modelos Animais de Doenças , Feminino , Granuloma/induzido quimicamente , Granuloma/imunologia , Granuloma/patologia , Masculino , Mesentério/imunologia , Mesentério/patologia , Camundongos , Peritônio/imunologia , Peritônio/patologia , Fagócitos/imunologia , Fagócitos/patologia , Especificidade da EspécieRESUMO
OBJECTIVE: To observe the effect of electroacupuncture (EA) on the expression of autophagy mediated PI3K/AKT pathway and sex hormones in polycystic ovary syndrome (PCOS) rats, so as to explore the mechanisms underlying improvement of PCOS anovulatory infertility. METHODS: Six-week-old female Sprague-Dawley rats were randomly divided into control, model and EA groups (n=10 per group). The PCOS model was established by gavage of Letrozole solution (1.0 mg/kg), once daily for 21 consecutive days. Then, EA (2 Hz, 1 mA) was applied to the "Sanyinjiao"(SP6) and "Taichong"(LR3) for 20 min, once a day for 14 successive days. The morphological changes of the ovary were observed after H.E. staining. The contents of serum androgen (T), luteinizing hormone (LH) and anti-mullerian hormone (AMH) were detected by ELISA. The expression levels of phosphatidylinositol 3-kinase (PI3K), protein kinase B (AKT) and microtubule-associated protein 1 light chain 3 (LC3) in the ovary tissues were detected by Western blot. RESULTS: Compared with the control group, the levels of serum T, LH and AMH contents, and ovarian LC3 â ¡ expression and the ratio of LC3 â ¡/â were significantly increased (P<0.05), and the expression levels of PI3K and AKT proteins were considerably decreased in the model group (P<0.05). After EA intervention, the levels of serum T, LH, AMH, and the ratio of LC3 â ¡/â were considerably down-regulated (P<0.05), and those of PI3K and AKT were obviously up-regulated in the EA group (P<0.05). CONCLUSION: EA intervention can reduce serum T, LH and AMH contents, and the ratio of ovarian LC3 â ¡/â and up-regulate ovarian PI3K and AKT in PCOS rats, which may contribute to its effect in improving anovulatory infertility by suppressing autophagy.
Assuntos
Eletroacupuntura , Síndrome do Ovário Policístico , Pontos de Acupuntura , Animais , Autofagia , Feminino , Hormônios Esteroides Gonadais , Fosfatidilinositol 3-Quinases , Ratos , Ratos Sprague-DawleyRESUMO
OBJECTIVE: To observe the effect of electroacupuncture(EA) on proangiogenesis process and protein turn-over in a mouse model of sarcopenia, so as to explore its potential molecular mechanism anti-aging. METHODS: Fourteen 30-week-old male SAMP8 mice were randomly divided into a model group (n=7) and an EA group (n=7). Seven anti-rapidly aging SAMR1 mice of the same age were used as the control group (n=7). EA (1 mA, 4 Hz) was applied to bilateral "Zusanli"(ST36) and "Yanglingquan"(GB34) for 20 minutes each time once a day, 6 times a week for 4 weeks. The exhausted running platform was used to test the sports function. Gastrocnemius muscle mass and relative ratio of gastrocnemius muscle mass to body mass were measured. HE staining and transmission electron microscope were used to observe the morphology, and the cross-sectional area of gastrocnemius muscle was calculated. Relative protein expressions of protein kinase B (AKT) , phosphorylated (p) -AKT, mammalian target of rapamycin (mTOR) , p-mTOR, p70 ribosomal protein S6 kinase (p70S6K) , p-p70S6K,hypoxia inducible factor-1α (HIF-1α) and relative mRNA expressions of HIF-1α, vascular endothelial growth factor A (VEGF-A) , muscle RING finger-1 (MuRF-1) and muscle atrophy F-box (MAFbx) were detected by Western blot and real-time fluorescence quantitative PCR, seperatively. RESULTS: Compared with the control group, the running time and distance, body mass and gastrocnemius mass, and the ratio of gastrocnemius mass to body mass decreased(P<0.01, P<0.05), cross-sectional area of gastrocnemius, related protein expression of p-AKT,p-mTOR, p-p70S6K and HIF-1α, mRNA expression of HIF-1α and VEGF-A decreased (P<0.01), while mRNA expression of MuRF1 and MAFbx increased (P<0.01) in the model group. Following EA intervention, the running time and distance, body mass and gastrocnemius mass and the ratio of gastrocnemius mass to body mass increased (P<0.05), cross-sectional area of gastrocnemius, related protein expression of p-AKT,p-mTOR, p-p70S6K and HIF-1α, mRNA expression of HIF-1α and VEGF-A were significantly up-regulated (P<0.01), mRNA expression of MuRF1 and MAFbx down-regulated (P<0.01, P<0.05) in the EA group compared with the model group. CONCLUSION: EA may delay the aging muscle atrophy in mice by regulating the gastrocnemius muscle's proangiogenesis process and protein turnover.
Assuntos
Eletroacupuntura , Sarcopenia , Animais , Masculino , Camundongos , Atrofia Muscular , Ratos , Ratos Sprague-Dawley , Fator A de Crescimento do Endotélio Vascular/genéticaRESUMO
Carbon dots (CDs) have been widely used for the preparation of multifunctional probes by conjugation with organic fluorescent dyes. However, the effect of organic fluorescent dyes on CDs still remains poorly understood. Herein, the effect of fluorescein isothiocyanate (FITC) on CDs was explored by spectroscopic techniques at pH5.1, 7.0 and 9.0. The fluorescent intensity of CDs was found to be quenched gradually after mixing directly with different concentrations of FITC, but the fluorescent lifetime of CDs remained unchanged. According to the results of UV-vis absorption spectra and fluorescent lifetime measurements, a pH-dependent inner filter effect (IFE) between CDs and FITC was proposed. However, the fluorescent lifetime of CDs deceased after their conjugation with FITC, implying the fluorescence resonance energy transfer (FRET) between CDs and FITC. This study has revealed two different effects of FITC on CDs with varying pH values and provided useful theoretical guidelines for further research on the interaction between other nanoparticles and fluorophores.