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BACKGROUND: Right ventricular (RV) fibrosis is an important pathological change that occurs during the development of right heart failure (RHF) induced by pulmonary hypertension (PH). Dapagliflozin (DAPA), a sodium-glucose cotransporter 2 (SGLT2) inhibitor, has been shown to play a major role in left heart failure, but it is unclear whether it has a positive effect on RHF. This study aimed to clarify the effect of DAPA on PH-induced RHF and investigate the underlying mechanisms. METHODS: We conducted experiments on two rat models with PH-induced RHF and cardiac fibroblasts (CFs) exposed to pathological mechanical stretch or transforming growth factor-beta (TGF-ß) to investigate the effect of DAPA. RESULTS: In vivo, DAPA could improve pulmonary hemodynamics and RV function. It also attenuated right heart hypertrophy and RV fibrosis. In vitro, DAPA reduced collagen expression by increasing the production of matrix metalloproteinase 2 (MMP2) and matrix metalloproteinase 9 (MMP9). Additionally, DAPA was found to reduce reactive oxygen species (ROS) levels in CFs and the right heart in rats. Similar to DAPA, the ROS scavenger N-acetylcysteine (NAC) exerted antifibrotic effects on CFs. Therefore, we further investigated the mechanism by which DAPA promoted collagen degradation by reducing ROS levels. CONCLUSIONS: In summary, we concluded that DAPA ameliorated PH-induced structural and functional changes in the right heart by increasing collagen degradation. Our study provides new ideas for the possibility of using DAPA to treat RHF.
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Background: Shear stress-induced Dickkopf-1 (DKK1) secretion by endothelial cells (ECs) promotes EC dysfunction and accelerates atherosclerosis (AS). However, the paracrine role of endothelial DKK1 in modulating adjacent smooth muscle cells (SMCs) in atherosclerosis remains unclear. This study investigated the role of EC-secreted DKK1 in SMC-derived foam cell formation under shear stress, in vitro and in vivo. Methods: Parallel-plate co-culture flow system was used to explore the cellular communication between ECs and SMCs under shear stress in vitro. Endothelium-specific knockout of DKK1 (DKK1ECKO/APOE-/-) and endothelium-specific overexpression of DKK1 (DKK1ECTg) mice were constructed to investigate the role of endothelial DKK1 in atherosclerosis and SMC-derived foam cell formation in vivo. RNA sequencing (RNA-seq) was used to identify the downstream targets of DKK1. Reverse transcription quantitative polymerase chain reaction (RT-qPCR), western blot, coimmunoprecipitation (Co-IP) assays and chromatin immunoprecipitation (ChIP) experiments were conducted to explore the underlying regulatory mechanisms. Results: DKK1 is transcriptionally upregulated in ECs under conditions of low shear stress, but not in co-cultured SMCs. However, DKK1 protein in co-cultured SMCs is increased via uptake of low shear stress-induced endothelial DKK1, thereby promoting lipid uptake and foam cell formation in co-cultured SMCs via the post-translational upregulation of scavenger receptor-A (SR-A) verified in parallel-plate co-culture flow system, DKK1ECKO and DKK1ECTg mice. RNA sequencing revealed that DKK1-induced SR-A upregulation in SMCs is dependent on Ubiquitin-specific Protease 53 (USP53), which bound to SR-A via its USP domain and cysteine at position 41, exerting deubiquitination to maintain the stability of the SR-A protein by removing the K48 ubiquitin chain and preventing proteasomal pathway degradation, thereby mediating the effect of DKK1 on lipid uptake in SMCs. Moreover, DKK1 regulates the transcription of USP53 by facilitating the binding of transcription factor CREB to the USP53 promoter. SMC-specific overexpression of USP53 via adeno-associated virus serotype 2 vectors in DKK1ECKO/APOE-/- mice reversed the alleviation of atherosclerotic plaque burden, SR-A expression and lipid accumulation in SMCs within plaques resulting from DKK1 deficiency. Conclusions: Our findings demonstrate that, endothelial DKK1, induced by pathological low shear stress, acts as an intercellular mediator, promoted the foam cell formation of SMCs. These results suggest that targeted intervention with endothelial DKK1 may confer beneficial effects on atherosclerosis.
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Aterosclerose , Células Espumosas , Peptídeos e Proteínas de Sinalização Intercelular , Miócitos de Músculo Liso , Animais , Aterosclerose/metabolismo , Camundongos , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Peptídeos e Proteínas de Sinalização Intercelular/genética , Células Espumosas/metabolismo , Miócitos de Músculo Liso/metabolismo , Células Endoteliais/metabolismo , Humanos , Ubiquitinação , Masculino , Técnicas de Cocultura , Camundongos Knockout , Proteases Específicas de Ubiquitina/metabolismo , Proteases Específicas de Ubiquitina/genética , Camundongos Endogâmicos C57BLRESUMO
AIM: To compare endovascular coiling and surgical clipping for the evaluation of clinical outcomes in patients with unruptured intracranial aneurysms. MATERIAL AND METHODS: We searched MEDLINE, EMBASE, the Cochrane Library and three Chinese domestic electronic databases, namely, Wanfang, CNKI and VIP for studies published between January 1990 and January 2018. We included controlled clinical studies comparing clinical outcomes between surgical clipping and endovascular coiling treatments. Two researchers extracted the data and assessed the quality of the studies, and a meta-analysis was performed using RevMan 5 software. RESULTS: We analysed a total of 23 controlled clinical studies including 117,796 cases. Meta-analysis demonstrated similar ischaemia rates between clipping and coiling with an odds ratio [OR] of 1.36 (95% CI: 0.77?2.40). The occlusion rate and bleeding risk were higher with clipping than coiling; the pooled ORs were 5.31 (95% CI: 3.07?9.19) and 2.39 (95% CI: 1.82?3.13), respectively. In addition, clipping resulted in a longer hospital stay (OR = 2.90, 95% CI: 2.14?3.65) than coiling did. Patients who underwent clipping had a higher short-term mortality (OR = 1.99, 95% CI: 1.70?2.33) and neurological deficit rate (OR = 2.05, 95% CI: 1.73? 2.44) compared with those who underwent coiling. However, 1 year mortality and deficit rate were similar for both clipping and coiling, with pooled ORs of 0.75 (95% CI: 0.41?1.38) and 0.94 (95% CI: 0.53?1.67), respectively. Funnel plots did not demonstrate a publication bias, with the exception of ischaemic outcome, and sensitivity analysis showed consistent results. CONCLUSION: Our study demonstrates that coiling is associated with a lower rate of occlusion, shorter hospital stay, lower bleeding risk and lower short-term mortality and morbidity compared with clipping. In terms of ischaemic risk, 1 year mortality and morbidity, coiling and clipping bear a similar risk. In addition, we speculate that surgical clipping may have a better outcome than endovascular coiling in the long term especially in young patients. Further research is needed to confirm our conclusion.
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Procedimentos Endovasculares , Aneurisma Intracraniano , Instrumentos Cirúrgicos , Humanos , Aneurisma Intracraniano/cirurgia , Aneurisma Intracraniano/terapia , Procedimentos Endovasculares/métodos , Resultado do Tratamento , Procedimentos Neurocirúrgicos/métodos , Embolização Terapêutica/métodos , Embolização Terapêutica/instrumentaçãoRESUMO
Photomorphogenesis is a light-dependent plant growth and development program. As the core regulator of photomorphogenesis, ELONGATED HYPOCOTYL 5 (HY5) is affected by dynamic changes in its transcriptional activity and protein stability; however, little is known about the mediators of these processes. Here, we identified PHOTOREGULATORY PROTEIN KINASE 1 (PPK1), which interacts with and phosphorylates HY5 in Arabidopsis, as one such mediator. The phosphorylation of HY5 by PPK1 is essential to establish high-affinity binding with B-BOX PROTEIN 24 (BBX24) and CONSTITUTIVE PHOTOMORPHOGENIC 1 (COP1), which inhibit the transcriptional activity and promote the degradation of HY5, respectively. As such, PPKs regulate not only the binding of HY5 to its target genes under light conditions but also HY5 degradation when plants are transferred from light to dark. Our data identify a PPK-mediated phospho-code on HY5 that integrates the molecular mechanisms underlying the regulation of HY5 to precisely control plant photomorphogenesis.
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Proteínas de Arabidopsis , Arabidopsis , Fatores de Transcrição de Zíper de Leucina Básica , Regulação da Expressão Gênica de Plantas , Luz , Proteínas de Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Arabidopsis/metabolismo , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/genética , Fatores de Transcrição de Zíper de Leucina Básica/metabolismo , Fatores de Transcrição de Zíper de Leucina Básica/genética , Fosforilação , Ubiquitina-Proteína Ligases/metabolismo , Ubiquitina-Proteína Ligases/genética , Proteínas Nucleares/metabolismo , Proteínas Nucleares/genética , Morfogênese/efeitos da radiação , Proteínas Quinases/metabolismo , Proteínas Quinases/genética , Proteínas RepressorasRESUMO
Background: Research has shown that carotid intima-media thickness (CIMT) could help to predict carotid plaque (CP) progression in patients with mild carotid stenosis. However, the debate continues as to the value of carotid intima thickness (CIT) in monitoring the development of CP in patients with severe carotid stenosis. This study sought to evaluate the relationships between CIT and the ultrasonic characteristics of CP and to analyze the value of CIT and the ultrasonic parameters of CP in assessing plaque vulnerability in advanced human carotid atherosclerosis. Methods: A total of 55 individuals who underwent carotid endarterectomy (CEA) were included in the study (mean age: 65±7 years; female: 9.1%). CIMT and CIT were examined at the common carotid artery (CCA). Plaque textural features, such as the gray-scale median (GSM), superb microvascular imaging (SMI) level, and total plaque area (TPA), were also identified. A Spearman correlation coefficient analysis was performed to examine the relationship between CIT and the ultrasonic parameters of CP. The CIT of various plaque types was compared. Receiver operating characteristic (ROC) curves were used to analyze the diagnostic values of the ultrasound characteristics to evaluate CP vulnerability. Results: The mean CIT of all the participants was 0.382±0.095 mm, the mean CIT of the participants with stable plaques was 0.328±0.031 mm, and the mean CIT of participants with vulnerable plaques was 0.424±0.106 mm (P<0.001). CIT was associated with the SMI level (Spearman's correlation coefficient: r=0.392, P=0.005), TPA (Spearman's correlation coefficient: r=0.337, P=0.012). Patients with thicker CIT had larger lipid cores, higher levels of plaque vulnerability, and more intraplaque hemorrhages (IPHs). The areas under the ROCs (AUCs) with 95% confidence interval (CI) for CIMT, CIT, the SMI level, the GSM, the TPA, and the combined model for identifying vulnerable plaques were 0.673 (0.533-0.793), 0.849 (0.727-0.932), 0.771 (0.629-0.879), 0.669 (0.529-0.790), 0.858 (0.738-0.938), and 0.949 (0.854-0.990), respectively. Conclusions: CIT was associated with both the histology and ultrasonic features of CP. CIT may be helpful in the detection of severe CP development.
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Vascular smooth muscle cells (VSMCs) are considered to be a crucial source of foam cells in atherosclerosis due to their low expression level of cholesterol exporter ATP-binding cassette transporter A1 (ABCA1) intrinsically. While the definite regulatory mechanisms are complicated and have not yet been fully elucidated, we previously reported that Dickkopf-1 (DKK1) mediates endothelial cell (EC) dysfunction, thereby aggravating atherosclerosis. However, the role of smooth muscle cell (SMC) DKK1 in atherosclerosis and foam cell formation remains unknown. In this study, we established SMC-specific DKK1-knockout (DKK1SMKO ) mice by crossbreeding DKK1flox/flox mice with TAGLN-Cre mice. Then, DKK1SMKO mice were crossed with APOE-/- mice to generate DKK1SMKO /APOE-/- mice, which exhibited milder atherosclerotic burden and fewer SMC foam cells. In vitro loss- and gain-of-function studies of DKK1 in primary human aortic smooth muscle cells (HASMCs) have proven that DKK1 prevented oxidized lipid-induced ABCA1 upregulation and cholesterol efflux and promoted SMC foam cell formation. Mechanistically, RNA-sequencing (RNA-seq) analysis of HASMCs as well as chromatin immunoprecipitation (ChIP) experiments showed that DKK1 mediates the binding of transcription factor CCAAT/enhancer-binding protein delta (C/EBPδ) to the promoter of cytochrome P450 epoxygenase 4A11 (CYP4A11) to regulate its expression. In addition, CYP4A11 as well as its metabolite 20-HETE-promoted activation of transcription factor sterol regulatory element-binding protein 2 (SREBP2) mediated the DKK1 regulation of ABCA1 in SMC. Furthermore, HET0016, the antagonist of CYP4A11, has also shown an alleviating effect on atherosclerosis. In conclusion, our results demonstrate that DKK1 promotes SMC foam cell formation during atherosclerosis via a reduction in CYP4A11-20-HETE/SREBP2-mediated ABCA1 expression.
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Aterosclerose , Células Espumosas , Humanos , Animais , Camundongos , Músculo Liso Vascular , Sistema Enzimático do Citocromo P-450 , Fatores de Transcrição , Aterosclerose/genética , Apolipoproteínas E/genética , Citocromo P-450 CYP4A , Transportador 1 de Cassete de Ligação de ATP/genéticaRESUMO
A multi-wavelength bandstop filter is proposed and numerically demonstrated using the sum-frequency generation (SFG) process in a waveguide of periodically poled lithium niobate (PPLN). This proposed device achieves channels number reconfigurable, central filtering wavelength of each filtering channel independently tunable and extinction ratios (ERs) equalized via all-optical methods.
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BACKGROUND: Enhancer of zeste homolog 2 (EZH2) was recently found to play an important role in cardiovascular disease. However, the role of EZH2 in vascular remodeling induced by mechanical stretch is poorly understood. The aim of the present work was to investigate the role of EZH2 in regulating smooth muscle cell function through mechanical stretch assays and to explore the underlying mechanisms. METHODS: WT C57BL/6 J mice underwent sham surgery or abdominal aortic constriction. The level of EZH2 expression was determined by Western blotting and immunohistochemical staining. We demonstrated the thickness of vascular remodeling by HE staining. JASPAR was used to predict transcription factors that could affect EZH2. Chromatin immunoprecipitation was used to substantiate the DNAprotein interactions. Promoter luciferase assays were performed to demonstrate the activity of the transcription factors. RESULTS: We found that in vivo, AAC significantly reduced EZH2 protein levels in the thoracic aorta. Smooth muscle-specific overexpression of EZH2 was sufficient to attenuate the AAC-induced reduction in trimethylation of Lys-27 in histone 3 and thickening of the arterial media. Administration of GSK-J4 (an inhibitor of H3K27me3 demethylase) induced the same effects. In addition, we found that mechanical stretch regulated the expression of EZH2 through the Yes-associated protein (YAP)- transcriptional factor TEA domain 1 (TEAD) pathway. TEAD1 bound directly to the promoter of EZH2, and blocking the YAP-TEAD1 interaction inhibited EZH2 downregulation due to mechanical stretch. CONCLUSION: This study reveals that mechanical stretch downregulates EZH2 through the YAP-TEAD1 pathway, thereby aggravating smooth muscle cell apoptosis and vascular remodeling.
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Proteína Potenciadora do Homólogo 2 de Zeste , Remodelação Vascular , Animais , Apoptose , Proliferação de Células , Proteína Potenciadora do Homólogo 2 de Zeste/genética , Proteína Potenciadora do Homólogo 2 de Zeste/metabolismo , Histonas/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Músculo Liso Vascular/metabolismo , Miócitos de Músculo Liso/metabolismo , Transdução de Sinais , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Proteínas de Sinalização YAPRESUMO
PURPOSE: This study aimed to compare the effect of needle puncture and chondroitinase ABC (ChABC) injection on inducing intervertebral disc (IVD) degeneration (IVDD) in rabbits. METHODS: Sixteen New Zealand white rabbits were used in this study. Briefly, the rabbits were divided into four groups. In the annulus fibrosis (AF) needle puncture group, a 16-G needle was used to puncture the L5-6 and L6-7 IVDs, while in the sham group, these IVDs were not punctured. In the ChABC group, 30 µL 0.5 Unit/mL ChABC was injected into L5-6 and L6-7 IVDs using a 26-G needle, while in the vehicle group, these IVDs were injected with 30 µL phosphate-buffered saline (PBS). X-ray and MRI scans were performed at the 4th, 12th and 16th weeks postoperatively. Histological, immunohistochemical and biochemical analyses were performed at the 16th week postoperatively. RESULTS: Both needle puncture and ChABC successfully established IVDD in rabbits at 4th, 12th and 16th weeks, confirmed by X-ray and MRI scan. The progression of IVDD went in a time-dependent manner. The IVDD in the ChABC group was less severe than in the needle puncture group throughout the study. Aggrecan and type II collagen significantly decreased, while tumor necrosis factor-α and superoxide dismutase 2 increased in the needle puncture and ChABC groups, compared with the sham and PBS groups. CONCLUSIONS: Both AF needle puncture and ChABC injection can successfully induce IVDD in rabbits. Compared with ChABC injection, AF needle puncture can induce more severe IVDD.
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Condroitina ABC Liase , Degeneração do Disco Intervertebral , Disco Intervertebral , Animais , Coelhos , Agrecanas , Condroitina ABC Liase/efeitos adversos , Colágeno Tipo II , Modelos Animais de Doenças , Disco Intervertebral/patologia , Degeneração do Disco Intervertebral/diagnóstico por imagem , Degeneração do Disco Intervertebral/patologia , Fator de Necrose Tumoral alfaRESUMO
Following the publication of this paper, it was drawn to the Editors' attention by a concerned reader that certain of the panels shown for the cell invasion assays in Figs. 2C and 6C were strikingly similar to data appearing in different form in other articles by different authors. Owing to the fact that the contentious data in the above article had were already under consideration for publication prior to its submission to Oncology Reports, the Editor has decided that this paper should be retracted from the Journal. The authors were asked for an explanation to account for these concerns, but the Editorial Office did not receive a reply. The Editor apologizes to the readership for any inconvenience caused. [Oncology Reports 18: 16701678, 2017; DOI: 10.3892/or.2017.5815].
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Vascular calcification (VC) is a significant risk factor for cardiovascular mortality and morbidity in patients with atherosclerosis (AS), chronic kidney disease, and diabetes. Dickkopf1 (Dkk1) is a multifunctional secreted glycoprotein that has been explored as a novel potential antitumor target. Recently, Dkk1 was shown to be closely associated with AS development. However, the role of Dkk1 in VC remains elusive. In this study, we explored the role and molecular mechanisms of Dkk1 in VC based on a smooth muscle-specific Dkk1-knockout (Dkk1SMKO) mouse model. Our data indicated that Dkk1 expression was decreased under calcifying conditions and that Dkk1 overexpression alleviated high phosphate-induced vascular calcification. In vivo, smooth muscle Dkk1-specific knockout aggravated vascular calcification in mice. However, phospholipase D1 (PLD1) overexpression partially weakened the protective effect of Dkk1 against vascular calcification. Mechanistically, Dkk1 slowed vascular calcification by promoting the degradation of PLD1 via the regulating autophagosome formation and maturation. In conclusion, we found that Dkk1 could alleviate vascular calcification by regulating the degradation of PLD1.
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Peptídeos e Proteínas de Sinalização Intercelular , Fosfolipase D , Insuficiência Renal Crônica , Calcificação Vascular , Animais , Camundongos , Miócitos de Músculo Liso/patologia , Fosfolipase D/metabolismo , Calcificação Vascular/genética , Calcificação Vascular/prevenção & controle , Camundongos Knockout , Peptídeos e Proteínas de Sinalização Intercelular/genéticaRESUMO
BACKGROUND: Pulmonary hypertension (PH) is a cancer-like proliferative disease, which has no curative treatment options. The dysfunction of pulmonary artery endothelial cells plays a key role in PH. DKK1 (Dickkopf 1) is a secretory glycoprotein that exerts proproliferative effects on tumor cells. In the present study, we aimed to identify the role and underlying mechanism of DKK1 in the development of PH, which still remain unclear. METHODS AND RESULTS: We found endothelial DKK1 expression was upregulated in serum and lung tissues obtained from patients with PH, mice with hypoxia-induced PH, and human pulmonary artery endothelial cells cultured under hypoxic conditions. Endothelium-specific DKK1-knockout (DKK1ECKO) mice significantly ameliorated hypoxia+Sugen5416 and hypoxia-induced PH. More importantly, neutralizing anti-DKK1 antibody treatment significantly attenuated established hypoxia+Sugen5416 PH. Results of proteome analysis of control and DKK1-knockdown human pulmonary artery endothelial cells identified a significantly differentially expressed protein, SHMT2 (serine hydroxymethyltransferase 2), a key metabolic enzyme in one-carbon metabolism, as a novel DKK1 target. DKK1 knockdown in human pulmonary artery endothelial cells cultured under hypoxic conditions decreased the cellular NADPH/NADP+ ratio, increased reactive oxygen species levels and the extent of mitochondrial DNA damage, and inhibited mitochondrial membrane hyperpolarization. In the context of this altered redox defense and mitochondrial disorder, DKK1 induced a proproliferative and antiapoptotic phenotype in endothelial cells. Furthermore, we confirmed that DKK1 regulated SHMT2 transcription through the AKT-Sp1 (specificity protein 1) signaling axis. CONCLUSIONS: Our data provide robust evidence and molecular explanations for the associations between DKK1, redox defense, mitochondrial disorders, and PH and reveal a novel target for PH treatment.
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Hipertensão Pulmonar , Animais , Células Endoteliais/metabolismo , Endotélio/metabolismo , Glicina Hidroximetiltransferase/genética , Glicina Hidroximetiltransferase/metabolismo , Humanos , Hipóxia/metabolismo , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Camundongos , Artéria Pulmonar/metabolismo , Fator de Transcrição Sp1/genética , Fator de Transcrição Sp1/metabolismo , Regulação para CimaRESUMO
OBJECTIVE: We sought to provide clinical evidence of the potential influence of ovariectomy (OVX) on intervertebral disk degeneration. METHODS: We retrospectively reviewed patients with a history of OVX who visited our hospital for lower back pain. In addition, 60 age-matched patients without OVX were randomly selected as control subjects. Next, the following demographic data were recorded and compared among groups: age, body mass index, duration of OVX, history of smoking, alcohol use, hypertension, diabetes, cardiocerebrovascular disease, hyperlipemia, osteoporosis, and degenerative spondylolisthesis. Next, the severity of lumbar disk degeneration, evaluated by the modified Pfirrmann grading system, was compared between groups. Data analyses were performed with SPSS 20.0. software. RESULTS: A total of 15 OVX (unilateral, n = 10; bilateral, n = 5) patients were included with a mean age of 62.40 ± 10.64. The average durations of OVX were 21.33 ± 9.24 years. There existed no remarkable intergroup differences in the demographic data (P > 0.05). Overall, the average Pfirrmann grading scores from L1/2 to L5/S1 presented as L1/2 < L2/3 < L3/4 ≤ L5/S1 ≤ L4/5, with no marked differences between groups (P > 0.05). Nevertheless, OVX groups displayed a relatively higher score at each level than non-OVX group. Moreover, the scores from L3/4 to L5/S1 were higher in the bilateral OVX group relative to the unilateral OVX group while they were equal at L1/2 and L2/3. CONCLUSIONS: Our findings demonstrated that OVX contributed to the progression of lumbar disk degeneration to some extent, but it appeared to be a long-term event.
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Degeneração do Disco Intervertebral/diagnóstico por imagem , Degeneração do Disco Intervertebral/epidemiologia , Vértebras Lombares/diagnóstico por imagem , Ovariectomia/tendências , Idoso , Estudos de Casos e Controles , Feminino , Humanos , Degeneração do Disco Intervertebral/etiologia , Dor Lombar/diagnóstico por imagem , Dor Lombar/epidemiologia , Dor Lombar/etiologia , Pessoa de Meia-Idade , Ovariectomia/efeitos adversos , Estudos RetrospectivosRESUMO
Dickkopf-1 (DKK1) was recently shown to play an important role in cardiovascular disease. The aim of this work was to assess the role of DKK1 in the regulation of smooth muscle cell function by mechanical stretch and the mechanisms underlying this process. Methods: Wild-type C57BL/6J mice were subjected to sham or abdominal aortic constriction (AAC) surgery. The expression level of DKK1 was examined by immunohistochemical staining and Western blotting. Analyses of DKK1 function in vascular smooth muscle cell (VSMC) proliferation and migration were performed. Transcriptome sequencing analysis was performed to identify the differentially expressed genes and pathways regulated by DKK1. Smooth muscle-specific Dkk1 knockout mice were used to confirm the function of DKK1 in vivo. Chromatin immunoprecipitation (ChIP) was used to confirm DNA-protein interactions. Promoter luciferase analysis was used to detect transcription factor activity. Results: We found that AAC significantly increased DKK1 protein levels in the thoracic aorta and coronary artery in vivo. In vitro, high-level stretch (18%) induced the expression of DKK1 in VSMCs. Knocking down DKK1 inhibited VSMC proliferation and migration under high-level stretch (18%). We identified ubiquitin-like containing PHD and RING finger domains 1 (UHRF1) as a target gene of DKK1. Knockdown of UHRF1 with small interfering RNAs partially reversed the regulatory effect of recombinant DKK1 on VSMCs. Specific deletion of DKK1 in VSMCs was sufficient to attenuate the AAC-induced upregulation of UHRF1, thickening of arterial media and increase in VSMC proliferation. Furthermore, we found that DKK1 regulated UHRF1 expression through the YAP-TEAD pathway. TEAD1 and TEAD4 bound directly to the promoter of UHRF1, and blocking the YAP-TEAD interaction inhibited UHRF1 upregulation due to DKK1. Conclusions: This study reveals that DKK1 mediates the mechanical stretch regulation of smooth muscle cell function by modulating UHRF1 expression through the YAP-TEAD pathway.
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Proteínas Estimuladoras de Ligação a CCAAT/metabolismo , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Músculo Liso Vascular/metabolismo , Fatores de Transcrição de Domínio TEA/metabolismo , Ubiquitina-Proteína Ligases/metabolismo , Proteínas de Sinalização YAP/metabolismo , Animais , Doenças Cardiovasculares/metabolismo , Proliferação de Células , Células Cultivadas , Regulação da Expressão Gênica , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Regulação para CimaRESUMO
BACKGROUND: Conventional high-throughput chemical screens in conjunction with genome-wide gene expression profiling proves to be successful in novel anti-cancer agent discovery and provides comprehensive insights into the mechanisms of action and off-target effects of single small-molecule compound. However, systematic evaluation on heterogeneous transcriptional responses of different cancer cell types to thousands of independent perturbations in a bioinformatics way is still limited. METHOD: Here, we introduce cancer transcriptome modifying potential (CTMP) which uses "Connectivity Score" to quantify and compare the effects of approved antineoplastic drugs on transcriptionally restoring dysregulated (both up- and down-) gene expressions at cancer state towards normal state. As a proof-of-concept, we applied this CTMP computational evaluation on > 10,000 small-molecule compounds using >200,000 Library of Integrated Network-based Cellular Signatures (LINCS) expression profiles generated upon 4 different cancer cell lines. We screened and proposed a candidate list of cancer transcriptome modifying therapeutics (CTMTs), among which the approved on-market drugs are further validated using GDSC drug sensitivity data, highlighting their potential to facilitate direct antineoplastic repositioning. RESULTS: In total, we calculated CTMPs of 85 on-market antineoplastic drugs and ~15,000 smallmolecule compounds using 253,813 transcriptomes across four cancer cell lines of lung, melanoma, prostate, and colon. Our results reveal that regardless of the chemical structure and targeted proteins majority of approved antineoplastic drugs present significant bilateral CTMPs across all 4 cancer cell lines. Bilateral CTMP-based systematic screen further indicates that candidate CTMTs are limited and most notably cancer-type specific. In particular, for each cancer cell type we proposed 3~5 CTMTs that are approved drugs with potent sensitivity data to support development in antineoplastic indications. CONCLUSION: Our work establishes CTMP to evaluate the antineoplastic property of small-molecule compounds and suggests CTMP-based systematic screen of cancer type-specific CTMTs as a feasible strategy in drug repositioning for precise anti-cancer purposes.
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Antineoplásicos/química , Biologia Computacional , Reposicionamento de Medicamentos , Ensaios de Triagem em Larga Escala , Neoplasias/genética , Bibliotecas de Moléculas Pequenas/química , Antineoplásicos/uso terapêutico , Perfilação da Expressão Gênica , Humanos , Neoplasias/tratamento farmacológico , Bibliotecas de Moléculas Pequenas/uso terapêuticoRESUMO
PURPOSE: To evaluate the efficacy of locking stand-alone cage (LSC) compared with anterior plate construct (APC) in anterior cervical discectomy and fusion (ACDF). METHODS: A comprehensive literature search was carried out in PubMed, Embase, Web of Science, and Cochrane Library to screen randomized controlled trials (RCTs) that directly compared LSC with APC in ACDF. The Cochrane Collaboration's tool was used for assessment of study quality. Data were analyzed with the Review Manager 5.3 software. RESULTS: A total of seven RCTs were included. The results revealed no significant differences between LSC and APC in ACDF regarding the fusion rate, Japanese Orthopaedic Association score, visual analogue scale score, neck disability index score, hospital stay, subsidence rate, cervical lordosis, segmental Cobb angle, and disc height. However, LSC was associated with a significantly shorter operation time, less blood loss, lower overall incidence of dysphagia, and lower adjacent-level ossification (ALO) rate compared with APC. CONCLUSION: In summary, LSC is not only a safe and effective device for ACDF but also has the advantages of significantly reduced operation time, blood loss, overall incidence of dysphagia, and ALO rate over APC. Therefore, LSC is a better alternative than APC for the patients undergoing ACDF procedures.
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Degeneração do Disco Intervertebral , Fusão Vertebral , Vértebras Cervicais/cirurgia , Discotomia , Humanos , Degeneração do Disco Intervertebral/cirurgia , Ensaios Clínicos Controlados Aleatórios como Assunto , Resultado do TratamentoRESUMO
BACKGROUND: Multidrug-resistant plasmids carrying replication genes have been widely present in various strains of Klebsiella pneumoniae. RepA and repB1 were found in plasmids belong to the IncFIB, but their detailed structural and genomic characterization was not reported yet. This is the first study that delivers structural and functional insights of repA- and repB1-carrying IncFIB plasmids. METHODS: Klebsiella pneumoniae strains A1705, 911021, and 1642 were isolated from the human urine samples and bronchoalveolar fluids collected from different hospitals of China. Antibacterial susceptibility and plasmid transfer ability were tested to characterize the resistant phenotypes mediated by the pA1705-qnrS, p911021-tetA, and p1642-tetA. The complete nucleotide sequences of these plasmids were determined through high-throughput sequencing technology and comparative genomic analyses of plasmids belong to the same incompatibility group were executed to extract the genomic variations and features. RESULTS: The pA1705-qnrS, p911021-tetA, and p1642-tetA are defined as non-conjugative plasmids, having two replication genes, repA and repB1 associated with IncFIB family, and unknown incompatible group, respectively. Comparative genomic analysis revealed that relatively small backbones of IncFIB plasmids integrated massive accessory module at one "hotspot" that was located between orf312 and repB1. These IncFIB plasmids exhibited the distinct profiles of accessory modules including one or two multidrug-resistant regions, many complete and remnant mobile elements comprising integrons, transposons and insertion sequences. The clusters of resistant genes were recognized in this study against different classes of antibiotics including ß-lactam, phenicol, aminoglycoside, tetracycline, quinolone, trimethoprim, sulfonamide, tunicamycin, and macrolide. It has been observed that all resistant genes were located in multidrug resistance regions. CONCLUSION: It is concluded that multidrug-resistant repA and repB1-carrying IncFIB plasmids are a key source to mediate the resistance through mobile elements among Klebsiella pneumoniae. Current findings provide a deep understanding of horizontal gene transfer among plasmids of the IncFIB family via mobile elements that will be utilized in further in vitro studies.
RESUMO
Background: In this study, it was aimed to investigate the clinical rehabilitation effect of lower-limb training on the patients that undergo oblique lumbar interbody fusion (OLIF) procedures. Methods: The eligible participants undergoing OLIF procedures between 01/2017 and 07/2019 were identified. All the patients underwent one-segment fusion operation (L3-4 or L4-5). Based on whether the participants received postoperative rehabilitation training, they were divided into two groups: intervention group and control group. Postoperatively, the participants in the intervention group were trained with lower-extremity rehabilitation exercise and maintained for three months. All participants got reexamined at the first postoperative week, the second postoperative week, the first postoperative month, and the third postoperative month (last follow-up). Comparisons were made in terms of the lower-extremity muscle force, visual analogue scale (VAS) score, lumbar JOA score, Oswestry disability index (ODI), the incidence of deep venous thrombosis (DVT), and patient satisfaction. Results: Seventy-seven participants in the intervention group (32 males and 45 females) and 82 in the control group (39 males and 43 females) were incorporated in this study. The median age of the participants was 57 years (39â¼73) in the intervention group and 54 years (35â¼71) in the control group. No statistical significance between the two groups was found (P > 0.05). ODI score was less in the intervention group as compared to the control group in the first week after surgery (P=0.029). VAS and JOA scores were better in the intervention group in the first two weeks after surgery (P < 0.05). DVT incidence in the intervention group was lower than the control group at final follow-up (P=0.037). Both group participants have achieved good grading in muscle force rehabilitation but no significant differences between the two groups. Additionally, satisfaction was higher in the intervention group than the control group. Conclusions: In summary, postoperative lower-extremity rehabilitation exercise can effectively accelerate patients' health recovery from the OLIF surgery and increase their satisfaction.
Assuntos
Terapia por Exercício/métodos , Recuperação de Função Fisiológica , Fusão Vertebral , Adulto , Idoso , Feminino , Humanos , Extremidade Inferior , Vértebras Lombares/cirurgia , Masculino , Pessoa de Meia-Idade , Satisfação do Paciente , Estudos RetrospectivosRESUMO
OBJECTIVES: To carry out a systematic review on the basis of overlapping meta-analyses that compare unilateral with bilateral pedicle screw fixation (PSF) in lumbar fusion to identify which study represents the current best evidence, and to provide recommendations of treatment on this topic. METHODS: A comprehensive literature search in PubMed, Embase, and the Cochrane Library databases was conducted to identify meta-analyses that compare unilateral with bilateral PSF in lumbar fusion. Only meta-analyses exclusively covering randomized controlled trials were included. Study quality was evaluated using the Oxford Levels of Evidence and Assessment of Multiple Systematic Reviews (AMSTAR) instrument. Then, the Jadad decision algorithm was applied to select the highest-quality study to represent the current best evidence. RESULTS: A total of 9 studies with Level II of evidence fulfilled the eligibility criteria and were included. The scores of AMSTAR criteria for them varied from 5 to 9 (mean 7.78). The current best evidence detected no significant differences between unilateral and bilateral PSF for short-segment lumbar fusion in the functional scores, length of hospital stay, fusion rate, and complication rate. However, unilateral PSF involved a remarkable decrease in operative time and blood loss but increase of cage migration when compared with bilateral PSF. CONCLUSIONS: According to this systematic review, unilateral PSF is an effective method of fixation for short-segment lumbar fusion, has the advantages of reduced operative time and blood loss over bilateral PSF, but increases the risk of cage migration.
Assuntos
Degeneração do Disco Intervertebral/cirurgia , Vértebras Lombares/cirurgia , Região Lombossacral/cirurgia , Parafusos Pediculares , Fusão Vertebral/métodos , Perda Sanguínea Cirúrgica , Humanos , Tempo de Internação , Duração da Cirurgia , Falha de Prótese , Fusão Vertebral/efeitos adversos , Resultado do TratamentoRESUMO
Nucleus pulposus cells (NPCs) play a vital role in maintaining the homeostasis of the intervertebral disc (IVD). Previous studies have discovered that NPCs exhibited malfunction due to cellular senescence during disc aging and degeneration; this might be one of the key factors of IVD degeneration. Thus, we conducted this study in order to investigate the altered biofunction and the underlying genes and pathways of senescent NPCs. We isolated and identified NPCs from the tail discs of young (2 months) and old (24 months) SD rats and confirmed the senescent phenotype through SA-ß-gal staining. CCK-8 assay, transwell assay, and cell scratch assay were adopted to detect the proliferous and migratory ability of two groups. Then, a rat Gene Chip Clariom™ S array was used to detect differentially expressed genes (DEGs). After rigorous bioinformatics analysis of the raw data, totally, 1038 differentially expressed genes with a fold change > 1.5 were identified out of 23189 probes. Among them, 617 were upregulated and 421 were downregulated. Furthermore, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were conducted and revealed numerous number of enriched GO terms and signaling pathways associated with senescence of NPCs. A protein-protein interaction (PPI) network of the DEGs was constructed using the Search Tool for the Retrieval of Interacting Genes (STRING) database and Cytoscape software. Module analysis was conducted for the PPI network using the MCODE plugin in Cytoscape. Hub genes were identified by the CytoHubba plugin in Cytoscape. Derived 5 hub genes and most significantly up- or downregulated genes were further verified by real-time PCR. The present study investigated underlying mechanisms in the senescence of NPCs on a genome-wide scale. The illumination of molecular mechanisms of NPCs senescence may assist the development of novel biological methods to treat degenerative disc diseases.