RESUMO
Detecting proteins at low concentrations in high-ionic-strength conditions by silicon nanowire field-effect transistors (SiNWFETs) is severely hindered due to the weakened signal, primarily caused by screening effects. In this study, aptamer as a signal amplifier, which has already been reported by our group, is integrated into SiNWFET immunosensors employing antigen-binding fragments (Fab) as the receptors to improve its detection limit for the first time. The Fab-SiNWFET immunosensors were developed by immobilizing Fab onto Si surfaces modified with either 3-aminopropyltriethoxysilane (APTES) and glutaraldehyde (GA) (Fab/APTES-SiNWFETs), or mixed self-assembled monolayers (mSAMs) of polyethylene glycol (PEG) and GA (Fab/PEG-SiNWFETs), to detect the rabbit IgG at different concentrations in a high-ionic-strength environment (150 mM Bis-Tris Propane) followed by incubation with R18, an aptamer which can specifically target rabbit IgG, for signal enhancement. Empirical results revealed that the signal produced by the sensors with Fab probes was greatly enhanced compared to the ones with whole antibody (Wab) after detecting similar concentrations of rabbit IgG. The Fab/PEG-SiNWFET immunosensors exhibited an especially improved limit of detection to determine the IgG level down to 1 pg/mL, which has not been achieved by the Wab/PEG-SiNWFET immunosensors.
Assuntos
Técnicas Biossensoriais , Nanofios , Animais , Imunoensaio , Limite de Detecção , Proteínas/análise , Coelhos , SilícioRESUMO
BACKGROUND: The purpose of this study is to determine the prevalence and the risk factors of DVT in end-stage OA patients. METHODS: From March 2015 to June 2017, 521 patients with knee degenerative osteoarthritis undergoing knee arthroplasty were enrolled; 458 patients (87.9%) were admitted for primary total knee arthroplasty and 63 patients (12.1%) were admitted for unicompartmental knee arthroplasty. Parameters were compared using χ2 or t-test for both the groups. Binary logistic regression analysis was used to determine risk factors. RESULTS: The incidence of preoperative DVT was 6.7% (n = 35). Age in preoperative DVT group was significantly more than the non-DVT group (72.54 ± 6.53 vs. 68.65 ± 7.35, P = 0.002). Preoperative D-dimer >0.5 µg/mL (P < 0.001) was also associated with preoperative DVT in knee osteoarthritis patients. The incidence increased with age significantly (2.17% in <65 years, 6.86% in ≥65 <75 years, and 12.26% in ≥75 years) (P = 0.008). Thus, age (P = 0.041, OR 1.075, 95% CI [1.002-1.110]) and D-dimer >0.5 µg/mL (P < 0.001, OR 4.441, 95% CI [1.942-10.153]) were the independent risk factors for preoperative DVT in knee osteoarthritis patients. CONCLUSIONS: The incidence of DVT in end-stage osteoarthritis was 6.7%. The results suggest that older people aged over 75 and D-dimer > 0.5 µg/mL were risk factors for DVT among patients admitted to the hospital for total knee arthroplasty. Instrumental screening should be encouraged, especially in subgroups at higher risk for preoperative DVT.
Assuntos
Osteoartrite do Joelho/epidemiologia , Trombose Venosa/epidemiologia , Adulto , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Artroplastia do Joelho , Biomarcadores/sangue , China/epidemiologia , Feminino , Produtos de Degradação da Fibrina e do Fibrinogênio/análise , Humanos , Incidência , Masculino , Pessoa de Meia-Idade , Osteoartrite do Joelho/diagnóstico , Osteoartrite do Joelho/cirurgia , Prevalência , Estudos Retrospectivos , Fatores de Risco , Trombose Venosa/diagnóstico por imagemRESUMO
BACKGROUND: Deep vein thrombosis (DVT) is common after total joint arthroplasty (TJA), and can cause the sequela of post-thrombotic syndrome (PTS), which is associated with decreased quality of life and increased treatment cost. The purpose of this study is to determine the incidence and risk factors for PTS in patients with DVT following primary unilateral total knee and hip arthroplasty. METHODS: We conducted this follow-up study involving patients developing DVT after primary unilateral total knee arthroplasty or total hip arthroplasty at our institution between April 2010 and March 2017. Each patient received a follow-up clinical interview regarding PTS-related symptoms and signs. We introduced demographic, clinical, and surgical data into the analysis to identify the risk factors for PTS. RESULTS: A total of 182 patients with postoperative DVT were enrolled with a mean follow-up time of 3.6 years. The incidence of PTS was 9.3% in patients developing DVT after TJA. Malignancy (P = .033), previous surgery in ipsilateral lower extremity (P = .013), and blood transfusion (P = .022) appeared to be the risk factors for PTS. CONCLUSION: We determined the incidence and risk factors for PTS in patients with DVT following TJA. Preventive measures should be used for patients at high risk of PTS.
Assuntos
Artroplastia de Quadril/efeitos adversos , Artroplastia do Joelho/efeitos adversos , Síndrome Pós-Trombótica/epidemiologia , Trombose Venosa/complicações , Adulto , Idoso , Idoso de 80 Anos ou mais , China/epidemiologia , Estudos de Coortes , Feminino , Seguimentos , Humanos , Incidência , Masculino , Pessoa de Meia-Idade , Síndrome Pós-Trombótica/etiologia , Qualidade de Vida , Fatores de RiscoRESUMO
OBJECTIVE: We investigated the role of endoplasmic reticulum stress (ERS) in silica-induced apoptosis in alveolar macrophages in vitro. METHODS: RAW264.7 cells were incubated with 200 µg/mL silica for different time periods. Cell viability was assayed by the MTT assay. Cell apoptosis was evaluated by DAPI staining, flow cytometry analysis, and Western blot analysis of caspase-3. Morphological changes in the endoplasmic reticulum were observed by transmission electron microscopy. The expression of ERS markers binding protein (BiP) and CCAAT-enhancer-binding protein homologous protein (CHOP) was examined by Western blotting and real-time PCR. As an inhibitor of ERS, 4-phenylbutyric acid (4-PBA) was used in the experiments. RESULTS: Silica exposure induced nuclear condensation and caspase-3 expression in RAW264.7 cells. The number of apoptotic cells increased after silica exposure in a time-dependent manner. Silica treatment induced expansion of the endoplasmic reticulum. In addition, the expression of BiP and CHOP increased in silica-stimulated cells. Furthermore, 4-PBA treatment inhibited silica-induced endoplasmic reticulum expansion and the expression of BiP and CHOP. Moreover, 4-PBA treatment attenuated nuclear condensation, reduced apoptotic cells, and downregulated caspase-3 expression in silica-stimulated cells. CONCLUSION: Silica-induced ERS is involved in the apoptosis of alveolar macrophages.
Assuntos
Apoptose/efeitos dos fármacos , Estresse do Retículo Endoplasmático/fisiologia , Dióxido de Silício/toxicidade , Animais , Butilaminas , Sobrevivência Celular/efeitos dos fármacos , Camundongos , Células RAW 264.7RESUMO
Bisphenol-A-glycidyldimethacrylate (BisGMA) is a frequently used monomer in dental restorative resins. However, BisGMA could leach from dental restorative resins after polymerization leading to inflammation in the peripheral environment. Wogonin, a natural flavone derivative, has several benefits, such as antioxidative, anti-inflammatory and neuroprotective properties. Pretreatment of macrophage RAW264.7 cells with wogonin inhibited cytotoxicity which is induced by BisGMA in a concentration-dependent manner. BisGMA induced apoptotic responses, such as redistribution of phosphatidylserine from the internal to the external membrane and DNA fragmentation, were decreased by wogonin in a concentration-dependent manner. In addition, BisGMA-induced genotoxicity, which detected by cytokinesis-blocked micronucleus and single-cell gel electrophoresis assays, were inhibited by wogonin in a concentration-dependent manner. Furthermore, wogonin suppressed BisGMA-induced activation of intrinsic caspase pathways, such as caspases-3 and -8. Parallel trends were observed in inhibition of caspase-3 and -8 activities, apoptosis, and genotoxicity. These results indicate wogonin suppressed the BisGMA-induced apoptosis and genotoxicity mainly via intrinsic caspase pathway in macrophages.
Assuntos
Antimutagênicos/farmacologia , Bis-Fenol A-Glicidil Metacrilato/toxicidade , Caspases/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Flavanonas/farmacologia , Sequestradores de Radicais Livres/farmacologia , Macrófagos/efeitos dos fármacos , Mutagênicos/toxicidade , Resinas Sintéticas/toxicidade , Animais , Fragmentação do DNA , Camundongos , Testes para Micronúcleos , Fosfatidilserinas/metabolismo , Células RAW 264.7 , Transdução de Sinais/efeitos dos fármacosRESUMO
Epidermal growth factor receptor tyrosine kinase inhibitors, including gefitinib, have been evaluated in patients with malignant gliomas. However, the molecular mechanisms involved in gefitinib-mediated anticancer effects against glioma are incompletely understood. In the present study, the cytostatic potential of gefitinib was demonstrated by the inhibition of glioma cell growth, long-term clonogenic survival, and xenograft tumor growth. The cytostatic consequences were accompanied by autophagy, as evidenced by monodansylcadaverine staining of acidic vesicle formation, conversion of microtubule-associated protein-1 light chain 3-II (LC3-II), degradation of p62, punctate pattern of GFP-LC3, and conversion of GFP-LC3 to cleaved-GFP. Autophagy inhibitor 3-methyladenosine and chloroquine and genetic silencing of LC3 or Beclin 1 attenuated gefitinib-induced growth inhibition. Gefitinib-induced autophagy was not accompanied by the disruption of the Akt/mammalian target of rapamycin signaling. Instead, the activation of liver kinase-B1/AMP-activated protein kinase (AMPK) signaling correlated well with the induction of autophagy and growth inhibition caused by gefitinib. Silencing of AMPK suppressed gefitinib-induced autophagy and growth inhibition. The crucial role of AMPK activation in inducing glioma autophagy and growth inhibition was further supported by the actions of AMP mimetic AICAR. Gefitinib was shown to be capable of reducing the proliferation of glioma cells, presumably by autophagic mechanisms involving AMPK activation.
Assuntos
Autofagia/efeitos dos fármacos , Glioma/tratamento farmacológico , Inibidores do Crescimento/farmacologia , Quinazolinas/farmacologia , Proteínas Quinases Ativadas por AMP/metabolismo , Aminoimidazol Carboxamida/análogos & derivados , Aminoimidazol Carboxamida/farmacologia , Animais , Proteínas Reguladoras de Apoptose/metabolismo , Linhagem Celular Tumoral , Gefitinibe , Glioma/metabolismo , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Proteínas Associadas aos Microtúbulos/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Ribonucleotídeos/farmacologia , Transdução de Sinais/efeitos dos fármacos , Serina-Treonina Quinases TOR/metabolismoRESUMO
Epithelial-mesenchymal transition (EMT) plays an important role in fibrotic diseases. We have previously showed that silica induces EMT in human bronchial epithelial cells (BECs); however, the underlying mechanism of silica-induced EMT is poorly understood. In the present study, we investigated the role of Snail in silica-induced EMT in human BECs in vitro. Human BECs were treated with silica at various concentrations and incubation times. Then MTT assay, western blot, electrophoretic mobility shift assay (EMSA), and small interfering RNA (siRNA) transfection were performed. We found that silica increased the expression and DNA binding activity of Snail in human BECs. SNAI siRNA inhibited the silica-induced expression of Snail. Moreover, SNAI siRNA upregulated the expression of epithelial marker E-cadherin, but attenuated the expression of mesenchymal marker α-smooth muscle actin and vimentin in silica-stimulated cells. These results suggest that Snail mediates the silica-induced EMT in human BECs.
Assuntos
Brônquios/efeitos dos fármacos , Células Epiteliais/efeitos dos fármacos , Transição Epitelial-Mesenquimal/efeitos dos fármacos , Dióxido de Silício/toxicidade , Fatores de Transcrição/metabolismo , Actinas/metabolismo , Western Blotting , Brônquios/citologia , Brônquios/metabolismo , Caderinas/metabolismo , Técnicas de Cultura de Células , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Ensaio de Desvio de Mobilidade Eletroforética , Células Epiteliais/citologia , Células Epiteliais/metabolismo , Humanos , Tamanho da Partícula , RNA Interferente Pequeno/genética , Fatores de Transcrição da Família Snail , Fatores de Transcrição/genéticaRESUMO
OBJECTIVE: Regular monitoring of serum potassium after a total joint arthroplasty (TJA) is a form of routine examination that can help detect abnormal serum potassium levels and reduce the incidences of adverse events that may occur on account of postoperative hypokalemia. Previous studies rarely discussed hypokalemia after joint replacement. In the present study, our primary goal was to investigate the incidence and possible risk factors of hypokalemia after a total hip and knee replacement procedure was performed. METHODS: This study included patients who underwent a unilateral total knee or hip arthroplasty in our department between April 2017 and March 2018. Serum potassium levels pre and post operation were collected and retrospectively analyzed. The differences in age, gender, body mass index (BMI), history of diseases, red blood cell (RBC), hemoglobin, hematocrit, glomerular filtration rate, ejection fraction, blood glucose, urine creatinine, urea nitrogen, intraoperative blood loss, operation time, drainage, preoperative potassium, surgery type, were compared between those patients diagnosed with hypokalemia and their non-hypokalemia at different times post surgery. Thereafter, the risk factors of postoperative hypokalemia patients were analyzed using statistical procedure multiple logistic regression model. RESULTS: The risk of hypokalemia after TJA was 53.1%, while, that on the first, third, and fifth day after operation was 12.5%, 40.7%, and 9.6% respectively. The serum potassium level on the first, third, and fifth postoperative days was 3.84 ± 0.32, 3.59 ± 0.34, and 3.80 ± 0.32 mmol/l, respectively. However, the level on the third day appeared to be the lowest (p = 0.015) of them all. The independent risk factors for hypokalemia after a total hip and knee replacement were the level of preoperative serum potassium concentration (p = 0.011), preoperative red blood cells counts (p = 0.027), and history of diabetes (p = 0.007). CONCLUSION: Regular monitoring of serum potassium concentration should be performed post TJA. We need to pay more attention to the patient's preoperative potassium levels along with their red blood cell counts especially in patients diagnosed with type 2 diabetes mellitus.
Assuntos
Artroplastia de Quadril , Artroplastia do Joelho , Diabetes Mellitus Tipo 2 , Hipopotassemia , Humanos , Artroplastia do Joelho/efeitos adversos , Artroplastia de Quadril/efeitos adversos , Estudos Retrospectivos , Hipopotassemia/epidemiologia , Hipopotassemia/etiologia , Prevalência , Fatores de Risco , PotássioRESUMO
Rheumatoid arthritis (RA) is a chronic systemic autoimmune disease characterized by synovial inflammation and the production of autoantibodies. Previous studies have indicated an association between high-salt diets (HSD) and an increased risk of RA, yet the underlying mechanisms remain unclear. Macrophage pyroptosis, a pro-inflammatory form of cell death, plays a pivotal role in RA. In this study, we demonstrate that HSD exacerbates the severity of arthritis in collagen-induced arthritis (CIA) mice, correlating with macrophage infiltration and inflammatory lesions. Given the significant alterations observed in macrophages from CIA mice subjected to HSD, we specifically investigate the impact of HSD on macrophage responses in the inflammatory milieu of RA. In our in vitro experiments, pretreatment with NaCl enhances LPS-induced pyroptosis in RAW.264.7 and THP-1 cells through the p38 MAPK/NF-κB signaling pathway. Subsequent experiments reveal that Slc6a12 inhibitors and SGK1 silencing inhibit sodium-induced activation of macrophage pyroptosis and the p38 MAPK/NF-κB signaling pathway, whereas overexpression of the SGK1 gene counteracts the effect of sodium on macrophages. In conclusion, our findings verified that high salt intake promotes the progression of RA and provided a detailed elucidation of the activation of macrophage pyroptosis induced by sodium transportation through the Slc6a12 channel.
Assuntos
Artrite Reumatoide , Macrófagos , Proteínas Serina-Treonina Quinases , Piroptose , Animais , Camundongos , Artrite Reumatoide/metabolismo , Artrite Reumatoide/patologia , Macrófagos/metabolismo , Piroptose/efeitos dos fármacos , Proteínas Serina-Treonina Quinases/metabolismo , Proteínas Serina-Treonina Quinases/genética , Cloreto de Sódio/farmacologia , Células RAW 264.7 , Humanos , Masculino , Proteínas Imediatamente Precoces/metabolismo , Proteínas Imediatamente Precoces/genética , Artrite Experimental/metabolismo , Transdução de Sinais , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Camundongos Endogâmicos DBARESUMO
OBJECTIVE: A molecular technique based on quasispecies analysis for tracing postexposure HIV transmission was applied in an investigation of a possible case of HIV transmission after blood transfusion. METHODS: Sixteen plasma specimens were collected from 3 HIV infections (T1-T3) involved in a possible HIV transmission chain and 13 HIV/AIDS (C1-C13) controls. The RNAs were extracted and then amplified by RT-PCR, the PCR products were cloned and sequenced.BioEdit 6.0.7 and MEGA 4.0 software were used to analyze gene sequences, calculate gene dispersion ratio and construct phylogenetic tree. RESULTS: The sequences of 13 specimens were successfully obtained.The HIV strains from T1, T2 and T3 were CRF07_BC recombinants, those from 5 out of the 6 controls lived in the same city with T2 and T3 were CRF07_BC recombinants as well, while those from 4 controls living in the same city with T1 were CRF01_AE recombinants. Compared with the clone sequences from T1, the mean gene dispersion ratio of T2 was the least (2.0%), followed by C12 (2.8%) , T3 (2.9%) and others. The phylogenetic tree showed that all clones from T1, T2, T3 and C12 might cluster together,and implied that the direction of HIV transmission was from T3 to T2, and then to T1. CONCLUSION: The results support the possible epidemiological clue that HIV was transmitted from T3 to T2, and then to T1, indicating that molecular epidemiological investigation could provide more direct evidence for tracing postexposure HIV transmission.
Assuntos
Infecções por HIV/transmissão , HIV/genética , Reação Transfusional , Feminino , Infecções por HIV/epidemiologia , Infecções por HIV/genética , Humanos , Masculino , Epidemiologia Molecular , Filogenia , RNA Viral/genética , Análise de Sequência de RNARESUMO
Traditional herbal medicine Ligusticum wallichii Franchat (Chuan Xiong) is frequently prescribed and highly recommended to patients with stroke. Rodent studies have demonstrated the neuroprotective effects of its active component tetramethylpyrazine against post-stroke brain injury and highlighted its role in antioxidant, anti-inflammation, and anti-apoptosis activity. Using permanent cerebral ischemia in rats and oxygen/glucose deprivation and reoxygenation (OGDR) in rat primary neuron/glia cultures, this study sheds light on the role of mitochondria as crucial targets for tetramethylpyrazine neuroprotection. Tetramethylpyrazine protected against injury and alleviated oxidative stress, interleukin-1ß release, and caspase 3 activation both in vivo and in vitro. Reduction of mitochondrial biogenesis- and integrity-related proliferator-activated receptor-gamma coactivator-1 alpha, mitochondrial transcription factor A (TFAM), translocase of outer mitochondrial membrane 20, mitochondrial DNA, and citrate synthase activity, as well as activation of mitochondrial dynamics disruption-related Lon protease, dynamin-related protein 1 (Drp1) phosphorylation, stimulator of interferon genes, TANK-binding kinase 1 phosphorylation, protein kinase RNA-like endoplasmic reticulum kinase phosphorylation, eukaryotic initiation factor 2α phosphorylation, and activating transcription factor 4 were revealed in permanent cerebral ischemia in rats and OGDR in neuron/glia cultures. TMP alleviated those biochemical changes. Our findings suggest that preservation or restoration of mitochondrial dynamics and functional integrity and alleviation of mitochondria-oriented pro-oxidant, pro-inflammatory, and pro-apoptotic cascades are alternative neuroprotective mechanisms of tetramethylpyrazine. Additionally, mitochondrial TFAM and Drp1 as well as endoplasmic reticulum stress could be targeted by TMP to induce neuroprotection. Data of this study provide experimental base to support clinical utility and value of Chuan Xiong towards stroke treatment and highlight an alternative neuroprotective target of tetramethylpyrazine.
Assuntos
Isquemia Encefálica , Oxigênio , Ratos , Animais , Glucose , Isquemia Encefálica/tratamento farmacológico , Infarto Cerebral , Mitocôndrias/metabolismoRESUMO
Rheumatoid arthritis (RA) is a chronic systemic autoimmune disease. Ethanol consumption has been reported to reduce morbidity in RA patients, but the mechanism behind it remains unclear. Our results showed that Muribaculaceae was predominant in the gut microbiota of mice after ethanol treatment, and the levels of microbiota metabolite acetate were increased. Acetate reduced arthritis severity in collagen-induced arthritis (CIA) mice, which was associated with a decrease in the articular neutrophils and the myeloperoxidase-deoxyribonucleic acid complex in serum. Meanwhile, in vitro experiments confirmed that acetate affected neutrophil activity by acting on G-protein-coupled receptor 43, which reduced endoplasmic reticulum stress in neutrophils and inhibited neutrophil extracellular traps formation. Furthermore, exogenous acetate reversed CIA mice with exacerbated gut microbial disruption, further confirming that the effect of gut microbial metabolite acetate on neutrophils in vivo is crucial for the immune regulation. Our findings illuminate the metabolic and cellular mechanisms of the gut-joint axis in the regulation of autoimmune arthritis, and may offer alternative avenues to replicate or induce the joint-protective benefits of ethanol without associated detrimental effects.
Assuntos
Artrite Experimental , Artrite Reumatoide , Armadilhas Extracelulares , Humanos , Camundongos , Animais , Artrite Reumatoide/tratamento farmacológico , Artrite Reumatoide/metabolismo , Neutrófilos , Artrite Experimental/tratamento farmacológico , Artrite Experimental/metabolismo , Acetatos/metabolismoRESUMO
Obesity is associated with accumulation of inflammatory immune cells in white adipose tissue, whereas thermogenic browning adipose tissue is inhibited. Dietary fatty acids are important nutritional components and several clinical and experimental studies have reported beneficial effects of docosahexaenoic acid (DHA) on obesity-related metabolic changes. In this study, we investigated effects of DHA on hepatic and adipose inflammation and adipocyte browning in high-fat diet-induced obese C57BL/6J mice, and in vitro 3T3-L1 preadipocyte differentiation. Since visceral white adipose tissue has a close link with metabolic abnormality, epididymal adipose tissue represents current target for evaluation. A course of 8-week DHA supplementation improved common phenotypes of obesity, including improvement of insulin resistance, inhibition of macrophage M1 polarization, and preservation of macrophage M2 polarization in hepatic and adipose tissues. Moreover, dysregulated adipokines and impaired thermogenic and browning molecules, considered obesogenic mechanisms, were improved by DHA, along with parallel alleviation of endoplasmic reticulum (ER) stress, mitochondrial dysfunction, and mitochondrial DNA stress-directed innate immunity. During 3T3-L1 preadipocytes differentiation, DHA treatment decreased lipid droplet accumulation and increased the levels of thermogenic, browning, and mitochondrial biogenesis molecules. Our study provides experimental evidence that DHA mitigates obesity-associated inflammation and induces browning of adipose tissue in visceral epididymal adipose tissue. Since obesity is associated with metabolic abnormalities across tissues, our findings indicate that DHA may have potential as part of a dietary intervention to combat obesity.
Assuntos
Dieta Hiperlipídica , Ácidos Docosa-Hexaenoicos , Camundongos , Animais , Ácidos Docosa-Hexaenoicos/metabolismo , Camundongos Obesos , Dieta Hiperlipídica/efeitos adversos , Tecido Adiposo Marrom/metabolismo , Camundongos Endogâmicos C57BL , Obesidade/metabolismo , Adipócitos , Tecido Adiposo Branco/metabolismo , Inflamação/tratamento farmacológico , Inflamação/metabolismo , TermogêneseRESUMO
BACKGROUND: Tendon derived stem cells (TDSCs) have proven to be effective in tendon repair by secreting paracrine factors, which modulate the function of resident cells and inflammatory process. Exosomes, which are secreted from cells to mediate intercellular communication, may be used to treat tendon injuries. Here, we aimed to determine the effects of exosomes from TDSCs (TDSC-Exos) on tendon repair and to explore the underlying mechanism by investigating the role of microRNAs (miRNAs). METHODS: TDSC-Exos were isolated from TDSC conditioned medium. In vitro studies were performed to investigate the effects of TDSC-Exos on the proliferation, migration, cytoprotection, collagen production and tendon-specific markers expression in tenocytes. In order to determine the therapeutic effects of TDSC-Exos in vivo, we used a scaffold of photopolymerizable hyaluronic acid (p-HA) loaded with TDSC-Exos (pHA-TDSC-Exos) to treat tendon defects in the rat model. Subsequently, RNA sequencing and bioinformatic analyses were used to screen for enriched miRNAs in TDSC-Exos and predict target genes. The miRNA-target transcript interaction was confirmed by a dual-luciferase reporter assay system. In order to determine the role of candidate miRNA and its target gene in TDSC-Exos-regulated tendon repair, miRNA mimic and inhibitor were transfected into tenocytes to evaluate cell proliferation and migration. RESULTS: Treatment with TDSC-Exos promoted proliferation, migration, type I collagen production and tendon-specific markers expression in tenocytes, and also protected tenocytes from oxidative stress and serum deprivation. The scaffold of pHA-TDSC-Exos could sever as a sustained release system to treat the rat model of tendon defects. In vivo study showed that TDSC-Exos promoted early healing of injured tendons. Rats treated with TDSC-Exos had better fiber arrangement and histological scores at the injury site. Besides, the injured tendons treated with TDSC-Exos had better performance in the biomechanical testing. Therefore, the pHA-TDSC-Exos scaffold proved to facilitate tendon repair in the rat model. miR-144-3p was enriched in TDSC-Exos and promoted tenocyte proliferation and migration via targeting AT-rich interactive domain 1A (ARID1A). CONCLUSIONS: TDSC-Exos enhanced tenon repair through miR-144-3p-regulated tenocyte proliferation and migration. These results suggest that TDSC-Exos can serve as a promising strategy to treat tendon injuries.
Assuntos
Exossomos , MicroRNAs , Animais , Proliferação de Células/genética , Exossomos/genética , Exossomos/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , Ratos , Células-Tronco/metabolismo , Tendões/metabolismo , Tenócitos/metabolismoRESUMO
BACKGROUND: Thyroid hormones (THs) and thyroid-stimulating hormone (TSH) seem to show high potential in predicting the clinical death outcome of patients admitted to the intensive care unit (ICU). However, diverse studies on this topic are conflicting. METHODS: A search was conducted by two investigators involved in this research in the PubMed, Embase, and Cochrane databases (all last launched on July 12, 2021). The quality of the included studies was evaluated using the Newcastle-Ottawa Quality Assessment Scale (NOS). Subgroup analyses were performed to determine the sources of heterogeneity. Sensitivity and publication bias analyses were also assessed. RESULTS: A total of 27 studies (4970 participants) were included based on the eligibility criteria. Compared with survivors, nonsurvivors were found to have lower levels of THs (T3, T4, fT3, and fT4), whereas no significant difference was found in TSH levels (13 studies for T3: standardized mean differences [SMD], -0.78; 95% CI, -1.36 to -0.20; I2 = 96%; p = 0.008; 11 studies for T4: SMD = -0.79; 95% CI, -1.31 to -0.28; I2 =95%; p = 0.0002; 14 studies for fT3: SMD = -0.76; 95% CI, -1.21 to -0.32; I2 = 95%; p = 0.0008; 17 studies for fT4: SMD = -0.60; 95% CI, -0.99 to -0.22; I2 = 95%; p = 0.002; 20 studies for TSH: SMD = 0.00; 93% CI, -0.29 to 0.29; I2 = 93%; p = 0.98). CONCLUSION: Nonsurvivors were associated with lower levels of THs (T3, T4, fT3, and fT4) than survivors. THs show great application potential in predicting ICU patients' death outcomes and improving already widely used prognostic scores in the ICU (ie, Acute Physiological and Chronic Health Evaluation [APACHE] II and Therapeutic Intervention Scoring System).
Assuntos
Tireotropina , Tiroxina , Adulto , Hospitalização , Humanos , Unidades de Terapia Intensiva , Hormônios TireóideosRESUMO
The role of NOD-like receptor protein 3 (NLRP3)-mediated macrophages pyroptosis in acute lung injury (ALI) is well-established. Quercetin (Que) is a natural bioflavonoid compound with anti-inflammatory and antioxidative properties that reportedly inhibits the NLRP3 inflammasome in sepsis-induced organ dysfunctions such as ALI. However, the mechanism underlying the inhibitory effect of quercetin on NLRP3 activation remains unclear. In this study, we established an endotoxin-induced ALI mouse model with an in vitro LPS challenge. We demonstrated that the administration of quercetin could significantly reduce pulmonary injury and decrease the production of pro-inflammatory cytokines. Moreover, we found that quercetin could inhibit the activation of the NLRP3 inflammasome by suppressing the nuclear accumulation of PKM2 and increasing SIRT1 levels. Importantly, treatment with SRT1720 (a specific SIRT1 activator) could inhibit the nuclear accumulation of PKM2 and the activation of NLRP3. Besides, preventing PKM2 dimerization with ML265 yielded an anti-inflammatory effect, similar to findings observed for SRT1720. In addition, we found that SIRT1 silencing or inhibition by EX527 could increase NLRP3 activation and nuclear accumulation of PKM2 and override quercetin-mediated anti-inflammatory activity. These findings indicated that quercetin could downregulate NLRP3 inflammasome activation by inhibiting the nuclear accumulation of PKM2 and upregulating SIRT1 expression, expanding the treatment landscape for ARDS.
Assuntos
Lesão Pulmonar Aguda , Inflamassomos , Animais , Camundongos , Lesão Pulmonar Aguda/induzido quimicamente , Lesão Pulmonar Aguda/tratamento farmacológico , Lesão Pulmonar Aguda/prevenção & controle , Anti-Inflamatórios/uso terapêutico , Inflamassomos/metabolismo , Lipopolissacarídeos/toxicidade , Camundongos Endogâmicos C57BL , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Quercetina/farmacologia , Quercetina/uso terapêutico , Sirtuína 1RESUMO
Plumbagin, a natural bicyclic naphthoquinone, has diverse pharmacological properties and biological benefits against a number of disorders, including liver disease. Though plumbagin's hepatoprotective potential attracts attention, currently no experimental evidence exists on its effectiveness against cholestatic liver injury. The present study investigated its hepatoprotection in the rat model of extrahepatic cholestasis using Bile Duct Ligation (BDL). We found that daily plumbagin supplementation protected the liver from cholestatic damage. Hepatoprotective actions of plumbagin were accompanied by reduction of Transforming Growth Factor ß1 (TGF-ß1)/Smad, High Mobility Group Box-1 (HMGB1)/Toll-Like Receptor-4 (TLR4), Hypoxia-Inducible Factor-1α (HIF-1α), Aryl Hydrocarbon Receptor (AhR), Heat Shock Protein 90 (HSP90), caveolin-1, NF-κB/AP-1, Dynamin Related Protein-1 (Drp1), malondialdehyde level, Interleukin-1ß (IL-1ß), p62/SQSTM1, and caspase 3 as well as increase of Farnesoid X Receptor (FXR), bile acid efflux transporters, glutathione, LC3-II, Beclin1, and nuclear NF-E2-Related Factor-2 (Nrf2) and Transcription Factor EB (TFEB). The activation of nuclear Nrf2 caused by plumbagin correlated well with the improvement in bile acid retention, liver histology, serum biochemical, ductular reaction, mitochondrial dysfunction, oxidative stress, inflammation, apoptosis, impaired autophagy, and fibrosis, involving interplay of multiple intracellular signaling pathways. Plumbagin is likely a candidate drug to protect the liver from cholestatic damages. Despite the promising findings from this study, translational implication of plumbagin on cholestatic liver injury warrants further investigation.
Assuntos
Ductos Biliares , Colestase , Fígado , Naftoquinonas , Animais , Ácidos e Sais Biliares/metabolismo , Ductos Biliares/patologia , Ductos Biliares/cirurgia , Colestase/complicações , Modelos Animais de Doenças , Ligadura , Fígado/efeitos dos fármacos , Fígado/metabolismo , Fígado/patologia , Fator 2 Relacionado a NF-E2/metabolismo , Naftoquinonas/farmacologia , Naftoquinonas/uso terapêutico , RatosRESUMO
18ß-Glycyrrhetinic acid is a nutraceutical agent with promising hepatoprotective effects. Its protective mechanisms against cholestatic liver injury were further investigated in a rodent model of extrahepatic cholestasis caused by Bile Duct Ligation (BDL) in rats. The daily oral administration of 18ß-Glycyrrhetinic acid improved liver histology, serum biochemicals, ductular reaction, oxidative stress, inflammation, apoptosis, impaired autophagy, and fibrosis. 18ß-Glycyrrhetinic acid alleviated the BDL-induced hepatic and systemic retention of bile acids, matrix-producing cell activation, hepatic collagen deposition, Transforming Growth Factor beta-1/Smad activation, malondialdehyde elevation, glutathione reduction, High Mobility Group Box-1/Toll-Like Receptor-4 activation, NF-κB activation, inflammatory cell infiltration/accumulation, Interleukin-1ß expression, Signal Transducer and Activator of Transcription-1 activation, Endoplasmic Reticulum stress, impairment autophagy, and caspase 3 activation. Conversely, the protein expression of Sirt1, Farnesoid X Receptor, nuclear NF-E2-Related Factor-2, Transcription Factor EB, bile acid efflux transporters, and LC3-II, as well as the protein phosphorylation of AMP-Activated Protein Kinase, was promoted in 18ß-Glycyrrhetinic acid-treated BDL rats. The hepatoprotective effects of 18ß-Glycyrrhetinic acid in the present investigation correlated well with co-activation and possible interactions among Sirt, FXR, and Nrf2. The concurrent or concomitant activation of Sirt1, FXR, and Nrf2 not only restored the homeostatic regulation of bile acid metabolism, but also alleviated oxidative stress, inflammation, apoptosis, impaired autophagy, and fibrosis.
RESUMO
OBJECTIVES: With the emergence of the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) B.1.1.7 lineage in the ongoing coronavirus disease 2019 (COVID-19) pandemic, Taiwan confronted a COVID-19 flare up in May 2021. Large-scale, accurate, affordable and rapid diagnostic tests such as the lateral flow assay can help to prevent community transmission, but their performance characteristics in real-world conditions and relevant subpopulations remain unclear. METHODS: The COVID-19 Antigen Rapid Test Kit (Eternal Materials, New Taipei City, Taiwan) was used in a high-throughput community testing site; the paired reverse transcription polymerase chain reaction (RT-PCR) results served as a reference for sensitivity and specificity calculations. RESULTS: Of 2096 specimens tested using the rapid antigen test, 70 (3.33%) were positive and 2026 (96.7%) were negative. This clinical performance was compared with the RT-PCR results. The sensitivity and specificity of the rapid antigen test were 76.39% [95% confidence interval (CI) 64.91-85.60%] and 99.26% (95% CI 98.78-99.58%), respectively, with high sensitivity in subjects with cycle threshold values ≤24. Further, the rapid antigen test detected the SARS-CoV-2 B.1.1.7 lineage effectively. CONCLUSIONS: Considering the short turnaround times and lower costs, this simple SARS-CoV-2 antigen detection test for rapid screening combined with RT-PCR as a double confirmatory screening tool can facilitate the prevention of community transmission during COVID-19 emergencies.
Assuntos
COVID-19 , SARS-CoV-2 , Antígenos Virais , Humanos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Sensibilidade e Especificidade , Taiwan/epidemiologiaRESUMO
OBJECTIVE: This study was to compare the performance of three HIV antibody confirmatory assay kits in confirming early HIV infection. METHODS: Five HIV antibody-positive plasma specimens were ten-fold serially diluted and then detected by ELISA. The above diluted specimens were detected with the following three HIV antibody confirmatory assay kits to analyze their sensitivity, including Wantai-RIBA (Recombinant immunoblot assay, Beijing Wantai Biological Pharmacy, China), MP-WB (HIV Blot 2.2 WB, MP Biomedicals Asia Pacific Pte. Ltd., Singapore) and INNO-LIA (INNO-LIA(TM) HIV I/II Score, Innogenetics N.V., Belgium), respectively. These kits were further used to detect 48 ELISA-reactive specimens from 11 sets of HIV seroconversion specimens (a total of 48 samples) which were previously detected as HIV antibody-positive by ELISA. RESULTS: When 5 samples were diluted to 100 fold, Wantai-RIBA still can detect them positive. Among the 48 HIV antibody-positive specimens detected with ELISA, the confirmation positive rate for Wantai-RIBA, MP-WB and INNO-LIA were 97.92% (47/48), 81.25% (39/48) and 91.67% (44/48), respectively. There was statistically significant difference between the confirmatory results of Wantai-RIBA and MP-WB (χ(2) = 6.13, P < 0.05), as well as between those of INNO-LIA and MP-WB (χ(2) = 5.48, P < 0.05); however, there was no statistically significant difference between those of Wantai-RIBA and INNO-LIA (χ(2) = 1.33, P > 0.05). For other six HIV seroconversion panels containing indeterminate specimens, the average seroconversion period of time for Wantai-RIBA, MP-WB and INNO-LIA were 0.7, 13.3 and 3.7 days, respectively. CONCLUSION: Compared with MP-WB, Wantai-RIBA and INNO-LIA could reduce the window period to confirm early HIV infection.