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BACKGROUND: To establish a method to induce Campylobacter jejuni colonization in the intestines of C57BL/6 mice through antibiotic-induced microbiome depletion. RESULTS: Fifty-four female C57BL/6 mice were divided into the normal, control, and experimental groups. The experimental group was administered intragastric cefoperazone sodium and sulbactam sodium (50 mg/mL) for 2 days; then, the experimental and control mice were intragastrically administered 200 µL C. jejuni, which was repeated once more after 2 days. Animal feces were collected, and the HipO gene of C. jejuni was detected using TaqMan qPCR from day 1 to day 14 after modeling completion. Immunofluorescence was used to detect intestinal C. jejuni colonization on day 14, and pathological changes were observed using hematoxylin and eosin staining. Additionally, 16S rDNA analyses of the intestinal contents were conducted on day 14. In the experimental group, C. jejuni was detected in the feces from days 1 to 14 on TaqMan qPCR, and immunofluorescence-labeled C. jejuni were visibly discernable in the intestinal lumen. The intestinal mucosa was generally intact and showed no significant inflammatory-cell infiltration. Diversity analysis of the colonic microbiota showed significant inter-group differences. In the experimental group, the composition of the colonic microbiota differed from that in the other 2 groups at the phylum level, and was characterized by a higher proportion of Bacteroidetes and a lower proportion of Firmicutes. CONCLUSIONS: Microbiome depletion induced by cefoperazone sodium and sulbactam sodium could promote long-term colonization of C. jejuni in the intestines of mice.
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Antibacterianos , Infecções por Campylobacter , Campylobacter jejuni , Cefoperazona , Fezes , Microbioma Gastrointestinal , Camundongos Endogâmicos C57BL , RNA Ribossômico 16S , Sulbactam , Animais , Campylobacter jejuni/efeitos dos fármacos , Campylobacter jejuni/crescimento & desenvolvimento , Feminino , Antibacterianos/farmacologia , Cefoperazona/farmacologia , Fezes/microbiologia , Infecções por Campylobacter/microbiologia , Camundongos , Microbioma Gastrointestinal/efeitos dos fármacos , Sulbactam/farmacologia , RNA Ribossômico 16S/genética , Intestinos/microbiologia , Colo/microbiologia , Colo/patologia , Modelos Animais de Doenças , Mucosa Intestinal/microbiologia , Mucosa Intestinal/efeitos dos fármacos , DNA Bacteriano/genética , DNA Ribossômico/genéticaRESUMO
We prepared BiOCl, BiO(ClBr), BiO(ClBrI), and BiO[ClBrI(CO3)0.5] materials using a simple coprecipitation method. It was found that adjusting the number of anions in the anion layer was conducive to adjusting the band structure of BiOX and could effectively promote the migration and separation of photogenerated carriers, thus improving the photocatalytic activity. We first selected methyl orange (MO) as the study pollutant and compared it with BiOCl, BiO(ClBr), and BiO(ClBrI). The first-order kinetic constants of MO degradation by BiO[ClBrI(CO3)0.5] increased by 90.3, 33.9, and 3.1 times, respectively. The photocatalytic degradation rate of methylene blue by BiO[ClBrI(CO3)0.5] was 89.5%, indicating the excellent photocatalytic performance of BiO[ClBrI(CO3)0.5]. The stability of BiO[ClBrI(CO3)0.5] was demonstrated through cyclic experiments and XRD analysis before and after the reaction. The photocatalytic degradation of MO by BiO[ClBrI(CO3)0.5] showed that h+ and 1O2 were the main active oxidizing species and â¢O2- was the secondary active substance. Overall, our work provides new ideas for the synthesis and degradation of organic pollutants by using two-dimensional anionic high-entropy materials.
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Wing polymorphism is an evolutionary feature found in a wide variety of insects, which offers a model system for studying the evolutionary significance of dispersal. In the wing-dimorphic planthopper Nilaparvata lugens, the insulin/insulin-like growth factor signaling (IIS) pathway acts as a 'master signal' that directs the development of either long-winged (LW) or short-winged (SW) morphs via regulation of the activity of Forkhead transcription factor subgroup O (NlFoxO). However, downstream effectors of the IIS-FoxO signaling cascade that mediate alternative wing morphs are unclear. Here we found that vestigial (Nlvg), a key wing-patterning gene, is selectively and temporally regulated by the IIS-FoxO signaling cascade during the wing-morph decision stage (fifth-instar stage). RNA interference (RNAi)-mediated silencing of Nlfoxo increase Nlvg expression in the fifth-instar stage (the last nymphal stage), thereby inducing LW development. Conversely, silencing of Nlvg can antagonize the effects of IIS activity on LW development, redirecting wing commitment from LW to the morph with intermediate wing size. In vitro and in vivo binding assays indicated that NlFoxO protein may suppress Nlvg expression by directly binding to the first intron region of the Nlvg locus. Our findings provide a first glimpse of the link connecting the IIS pathway to the wing-patterning network on the developmental plasticity of wings in insects, and help us understanding how phenotypic diversity is generated by the modification of a common set of pattern elements.
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Proteína Forkhead Box O1/metabolismo , Hemípteros/metabolismo , Proteínas de Insetos/metabolismo , Somatomedinas/metabolismo , Asas de Animais/crescimento & desenvolvimento , Animais , Proteína Forkhead Box O1/genética , Regulação da Expressão Gênica no Desenvolvimento/genética , Técnicas de Inativação de Genes , Ontologia Genética , Inativação Gênica , Hemípteros/genética , Hemípteros/crescimento & desenvolvimento , Sequenciamento de Nucleotídeos em Larga Escala , Proteínas de Insetos/genética , Íntrons , Fenótipo , Ligação Proteica , Interferência de RNA , Somatomedinas/genética , Análise Espaço-Temporal , Asas de Animais/metabolismoRESUMO
High beam quality 588â nm radiation was realized based on a frequency-doubled crystalline Raman laser. The bonding crystal of YVO4/Nd:YVO4/YVO4 was used as the laser gain medium, which can accelerate the thermal diffusion. The intracavity Raman conversion and the second harmonic generation were realized by a YVO4 crystal and an LBO crystal, respectively. Under an incident pump power of 49.2 W and a pulse repetition frequency of 50 kHz, the 588â nm power of 2.85 W was obtained with a pulse duration of 3â ns, corresponding to a diode-to-yellow laser conversion efficiency of 5.75% and a slope efficiency of 7.6%. Meanwhile, a single pulse's pulse energy and peak power were 57 µJ and 19â kW, respectively. The severe thermal effects of the self-Raman structure were overcome in the V-shaped cavity, which has excellent mode matching, and combined with the self-cleaning effect of `Raman scattering, the beam quality factor M2 was effectively improved, which was measured optimally to be Mx 2 = 1.207, and My 2 = 1.200, with the incident pump power being 49.2 W.
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Two modified V2C-MXene nanocoatings are prepared through different molecular weights of polyacrylic acid (polyacrylic acid with â¼4 50 000 is marked as LPAA, and polyacrylic acid with â¼4 000 000 is marked as HPAA) and two-dimensional V2C-MXene. Their properties are characterized using a ball-on-disc tribometer, three-dimensional white-light interferometry topography images, optical microscope, Raman spectrometer, focused ion beam/scanning electron microscope and high-resolution transmission electron microscope/energy dispersive X-ray spectrometer (HRTEM/EDS). As a result, an ultralow friction (µ ≈ 0.073 ± 0.024) and an ultralow wear (3.41 × 10-7 mm3 N-1 m-1 for ball scar, and 7.49 × 10-8 mm3 N-1 m-1 for disc track) are achieved for the LPAA@V2C vs. steel ball system tested under 4 N in the air through tribo-physicochemical interactions. During the rubbing process, the LPAA@V2C nanocoating is transferred onto counter-bodies to form mixed-phase lubricative tribofilms. Monitoring via a HRTEM/EDS, the mixed-phase lubricative tribofilms are found to be mainly composed of amorphous carbon phases containing O and V and layered nano-debris along the sliding surface. The tribofilm's stable structure is the key to realizing ultralow friction and ultralow wear through the LPAA modification. These findings disclose that MXene-based nanomaterials can be applied for material engineering and mechanical engineering under common working conditions.
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To explore the effects of Campylobacter jejuni lipopolysaccharide (Cj-LPS) on axonal injury in the spinal cord. Wistar rats were divided into the control (NC) group, model group (Cj-LPS), and LPS antibody group (Anti-LPS). Rats in the NC group were injected with a mixture of normal saline and complete Freund's adjuvant (CFA) while those in Cj-LPS group were injected with Cj-LPS, composed of LPS, CFA, and saline. Rats were sacrificed at 4th week and 6th week after injection, and hematoxylin and eosin (HE) staining was performed on the spinal cord sections. Real time-reverse transcription(RT-PCR) was used to detect mRNA expression of the axonal nutrition factor neurotrophin-3 (NT-3) with its receptor tropomyosin receptor kinase C (TrkC) and axon inhibitory factor of NogoA/NgR (Nogo receptor). The results indicated that Cj-LPS induce axonal injury in the rat spinal cord, decreased the mRNA expression of the axonal nutrition factor NT-3/TrkC, and increased the mRNA expression of the inhibitory factor NogoA/NgR. However, anti-LPS ameliorated axonal injury in the rat spinal cord induced by Cj-LPS.
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Campylobacter jejuni/metabolismo , Lipopolissacarídeos/farmacologia , Traumatismos da Medula Espinal/imunologia , Medula Espinal/efeitos dos fármacos , Animais , Anticorpos Antibacterianos/sangue , Anticorpos Antibacterianos/imunologia , Antígenos de Bactérias/imunologia , Modelos Animais de Doenças , Regulação da Expressão Gênica , Lipopolissacarídeos/imunologia , Masculino , RNA Mensageiro/metabolismo , Ratos , Ratos Wistar , Reação em Cadeia da Polimerase em Tempo Real , Cloreto de Sódio/farmacologia , Medula Espinal/patologia , Traumatismos da Medula Espinal/patologiaRESUMO
Background: Micronutrient levels play a critical role in epilepsy. This study investigates the impact of micronutrient levels on epilepsy via Mendelian randomization (MR). Methods: A two-sample MR framework evaluated the genetic association between 15 serum micronutrients and epilepsy phenotypes. The analysis included calcium, iron, zinc, selenium, copper, magnesium, potassium, folate, vitamins B6, B12, C, D, E, retinol, and carotene against all epilepsy, generalized epilepsy, childhood absence epilepsy (CAE), juvenile absence epilepsy (JAE), juvenile myoclonic epilepsy (JME), generalized tonic-clonic seizures alone and with spike-wave electroencephalography (GTCS), and various focal epilepsy phenotypes [with hippocampal sclerosis (HS), lesions other than HS, lesion-negative]. The random-effects inverse-variance weighted (IVW) model was the primary method used, supported by heterogeneity and pleiotropy assessments. Multivariable Mendelian randomization analyses (MVMR) were used to identify micronutrients that are significantly causally associated with different epilepsy subtypes and to confirm the most potential causal risk factors for these subtypes. Results: Zinc conferred an increased risk of focal epilepsy with HS (OR = 1.01; p = 0.045). Carotene was similarly linked to higher risks of lesion-negative cases (OR = 1.129; p = 0.037). Conversely, vitamin B6 was associated with reduced risks of focal epilepsy with HS (OR = 0.949; p = 0.020), and vitamin D was linked to decreased risks of both CAE (OR = 0.976, 95% CI: 0.959-0.993, p = 0.006) and JAE (OR = 0.986, 95% CI: 0.973-0.999, p = 0.032). These associations were robust, showing minimal heterogeneity and no evidence of pleiotropy across various sensitivity analyses. After adjustment using MVMR, significant causal relationships between vitamin D and both CAE and JAE remained. Furthermore, the causal relationship between zinc and vitamin B6 on focal epilepsy with HS became non-significant, while carotene shifted from a risk factor to a protective factor for focal epilepsy lesion-negative after adjusting for vitamin D. Conclusion: MR estimates provide robust evidence for the causal effects of vitamin D on reducing the risk of CAE, and JAE, which might provide alternative treatment strategies.
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In this work, N-graphyne is in situ coupled with BiOCl0.5Br0.5via a facile one-step sonochemical method. To our knowledge, both the synthesis strategy for BiOCl0.5Br0.5 and the N-graphyne/BiOCl0.5Br0.5 photocatalytic system are new developments. A collection of characterization methods is adopted to detect the morphologies, structures, and electronic and optical properties. The results demonstrate that wrinkle-like N-graphyne nanosheets successfully enwind around or on flower-like BiOCl0.5Br0.5 microspheres, which are regularly assembled by BiOCl0.5Br0.5 nanosheets. Compared with pristine BiOCl0.5Br0.5, N-graphyne/BiOCl0.5Br0.5 composites exhibit superior adsorption capacity and visible-light-driven photocatalytic degradation of levofloxacin. In particular, the optimal N-graphyne amount for ameliorating the photocatalytic performance of BiOCl0.5Br0.5 is ascertained. In addition, the good stable performance for photocatalysis is confirmed by four cycling experiments. The dominant active species is confirmed to be O2Ë- during photodegradation. The improved photocatalytic activity is attributed to the enhanced visible light response and the accelerated transfer/separation of photogenerated carriers by N-graphyne, which are verified using UV-vis absorption spectra, photocurrents, photopotentials, Nyquist plots, and Mott-Schottky curves. This study develops a new perspective for the synthesis and modification of BiOX solid solution, which can be used as an efficient photocatalyst.
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Herein, a collection of novel N-Ti3C2/BiOClxBr1-x composites are fabricated via a simple in-situ sonochemical process. Not only the preparation method for N-Ti3C2 but also the photocatalytic system of N-Ti3C2/BiOClxBr1-x are firstly developed. Multiple characterizations jointly demonstrate the successful fabrication of the composites. Compared to that of BiOClxBr1-x, the maximum improvements of 1.16, 1.25 and 1.26 folds are severally confirmed for the photocatalytic degradation of levofloxacin, Rhodamine B, and methylene blue over N-Ti3C2/BiOClxBr1-x composites. In addition, through radicals trapping tests, the primary active species in photocatalytic degradation process are verified to be O2-. Moreover, N-Ti3C2/BiOClxBr1-x composites also exhibit 1.18 and 1.14 times enhancements for NH3 production compared with that of BiOClxBr1-x with or without the presence of methanol, respectively. In addition, the maximum improvements of photo-current and photo-potential for BiOClxBr1-x are 1.29 and 1.86 folds with the introduction of N-Ti3C2, respectively. The enhanced photocatalytic activity of N-Ti3C2/BiOClxBr1-x composites is owing to the heightened light absorption, increased specific surface area, and accelerated separation of photoinduced carriers. Additionally, the stable photocatalytic properties of N-Ti3C2/BiOClxBr1-x are confirmed by three photocatalytic recycle tests on pollutant degradation and nitrogen reduction combined with X-ray diffraction patterns before and after three recycles. This study suggests that N-Ti3C2 is an efficient ornamentation for boosting photocatalytic activity ofBiOClxBr1-x, which can also be expanded as a promising modifier for other semiconductors.
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Herein, Nitrogen-doped graphyne/porous g-C3N4 composites are firstly in-situ synthesized via the ultrasound vibration of CaC2, triazine, and porous g-C3N4 in absolute ethanol. A variety of characterizations are performed to investigate the morphology, microstructure, composition, and electrical/optical features of the obtained composites, such as transmission electron microscopy, scanning electron microscopy, X-ray diffraction, Fourier transform infrared spectra, X-ray photoelectron spectroscopy, and so forth. It is found that N-doped graphyne with flexible folds lamellar structure is intimately attached to flake g-C3N4 in the as-prepared composites. An enlargement of 1.68 and 1.44 folds for the photocatalytic degradation of levofloxacin, rhodamine B, Methylene blue, and Tetracycline is realized by N-doped graphyne/g-C3N4 in comparison with that of pristine g-C3N4, respectively. In addition, the highest NH3 production rate attains 1.71 mmolâ gcat-1â h-1 for N-doped graphyne/g-C3N4, which is 5.89 times larger than that of g-C3N4 (0.29 mmolâ gcat-1â h-1). The improved mechanism of photocatalysis including higher photo-response and carrier separation rate is verified by transient photo-current, transient photo-potential, Mott-Schottky plots, Tafel plots, electrochemical impedance spectroscopy, turn-over frequency, photoluminescence spectra, and UV-vis diffuse absorption spectra, etc. Overall, the current study shows that N-doped graphyne synthesized from CaC2 and triazine is a useful decoration to modulate the photocatalytic features of g-C3N4, which can also be widely extended for in-situ modification of other photocatalysts.
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Herein, a series of N-doped Ti3C2/porous g-C3N4 composites are ultrasonically prepared from N-doped Ti3C2 and porous g-C3N4 under N2 atmosphere. The structure, morphology, and optical characteristics of the as-prepared composites are characterized by X-ray diffraction, transmission electron microscopy, scanning electron microscopy, X-ray photoelectron spectroscopy, UV-vis diffuse reflectance spectroscopy, etc. Moreover, photocatalytic measurements show that N-doped Ti3C2 is an excellent modifier for porous g-C3N4 to heighten its photocatalytic activity. Only 44.1% of rhodamine B can be degraded by the photocatalysis of pristine porous g-C3N4, while the photocatalytic degradation ratio of rhodamine B can reach up to 97.5% for the optimal N-doped Ti3C2 loading composites under visible light for 15 min. Moreover, the photocatalytic tests of N2 fixation confirm that the optimal composites show the highest production yield of NH4+ (11.8 µmol gcat-1 h-1), which is 2.11-folds more than that of porous g-C3N4 (5.6 µmol gcat-1 h-1). The reinforced photocatalytic properties are revealed to profit from the more photogenerated electrons and holes' separation, higher ability for light response, and more abundant active sites. This work develops the route for boosting the photocatalytic properties of porous g-C3N4 with N-doped Ti3C2.
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Due to the threat of lead pollution to health, environmental and food safety, developing simple and fast detection methods is highly required. Whereas, traditional single-mode probe suffers from limited application scenario. In this study, a colorimetric and fluorometric dual-mode probe for Pb2+ determination was constructed by using bifunctional G-quadruplex-hemin complex. In this dual-mode probe, enzyme strand and substrate strand of 8-17 DNAzyme are labeled with G-quadruplex-hemin complex and fluorophore, respectively. In the absence of Pb2+, the self-assembly of enzyme strand and substrate strand inhibits intrinsic mimic peroxidase of G-quadruplex-hemin complex by base-pairing, which also quench the fluorescence via in proximity effect. When the DNAzyme is activated by Pb2+, the quenched fluorescence is restored as well as the inherent peroxidase mimetic activity, leading to dual signal output. Under optimal conditions, this dual-mode probe exhibit a good linear relationship between logarithm of Pb2+ concentration and signal difference within the range from 1.5 nM to 20 nM and 0.5 nM to 10 nM for colorimetric and fluorescence mode, respectively. The detection limits for the corresponding mode were estimated to be 1.29 nM and 0.16 nM, respectively. This dual-mode probe also successfully applied for the spiked river water assay with satisfactory recovery in the range of 93.2 %-115.3 %. This work paves a new way for DNAzyme based dual-mode probe construction.
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Técnicas Biossensoriais , DNA Catalítico , Quadruplex G , Hemina , Chumbo , Colorimetria/métodos , Corantes , Peroxidases , Técnicas Biossensoriais/métodosRESUMO
Owing to advantage in high sensitivity and fast response, aptamer based electrochemical biosensors have attracted much more attention. However, inappropriate interfacial engineering strategy leads to poor recognition performance, which ascribe to the following factors of immobilized oligonucleotide strand including steric hindrance, interchain entanglement, and unfavorable conformation. In this work, we proposed a DNA tetrahedron based diblock aptamer immobilized strategy for the construction of label-free electrochemical biosensor. The diblock aptamer sequence is composite of T-rich anchor domain and recognition domain, where T-rich domain enabling anchored on the edge of DNA tetrahedron via Hoogsteen hydrogen bond at neutral condition. The DNA tetrahedron scaffold offers an appropriate lateral space for target recognition of diblock aptamer. More importantly, this trivalent aptamer recognition interface can be regenerated by simply adjusting the pH environment to alkaline, resulting in the dissociation of diblock aptamer. Under the optimum condition, proposed electrochemical aptasensor manifested a satisfied sensitivity for aminoglycosides antibiotic, kanamycin with a limit of detection of 0.69 nM, which is 45-fold lower than traditional Au-S immobilization strategy. Moreover, the proposed aptasensor had also successfully been extended to ampicillin detection by changing the sequence of recognition domain in diblock aptamer. This work paves a new way for the rational design of aptamer-based electrochemical sensor.
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Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Antibacterianos , Técnicas Biossensoriais/métodos , Aptâmeros de Nucleotídeos/química , DNA/química , Canamicina , Técnicas Eletroquímicas , Limite de Detecção , Ouro/químicaRESUMO
Optogenetics is a novel biotechnology widely used to precisely manipulate a specific peripheral sensory neuron or neural circuit. However, the use of optogenetics to assess the therapeutic efficacy of analgesics is elusive. In this study, we generated a transgenic mouse stain in which all primary somatosensory neurons can be optogenetically activated to mimic neuronal hyperactivation in the neuropathic pain state for the assessment of analgesic effects of drugs. A transgenic mouse was generated using the advillin-Cre line mated with the Ai32 strain, in which channelrhodopsin-2 fused to enhanced yellow fluorescence protein (ChR2-EYFP) was conditionally expressed in all types of primary somatosensory neurons (advillincre/ChR2+/+). Immunofluorescence and transdermal photostimulation on the hindpaws were used to verify the transgenic mice. Optical stimulation to evoke pain-like paw withdrawal latency was used to assess the analgesic effects of a series of drugs. Injury- and pain-related molecular biomarkers were investigated with immunohistofluorescence. We found that the expression of ChR2-EYFP was observed in many primary afferents of paw skin and sciatic nerves and in primary sensory neurons and laminae I and II of the spinal dorsal horns in advillincre/ChR2+/+ mice. Transdermal blue light stimulation of the transgenic mouse hindpaw evoked nocifensive paw withdrawal behavior. Treatment with gabapentin, some channel blockers, and local anesthetics, but not opioids or COX-1/2 inhibitors, prolonged the paw withdrawal latency in the transgenic mice. The analgesic effect of gabapentin was also verified by the decreased expression of injury- and pain-related molecular biomarkers. These optogenetic mice provide a promising model for assessing the therapeutic efficacy of analgesics in neuropathic pain.
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Breast cancer (BRCA) is a highly heterogeneous malignancy with an urgent need to build a proper model to predict its prognosis. Cuproptosis is a recently discovered form of cell death, mediated by protein fatty acylation and tightly associated with mitochondrial metabolism. The role of cuproptosis-related genes (CRGs) in BRCA remains to be explored. We aimed to investigate the applications of CRGs in BRCA prognosis in different clinical contexts, including chemotherapy and immunotherapy, via bioinformatics analysis of the messenger RNA profiles and clinical data obtained from public databases. Molecular subtyping of CRGs was performed through consistent clustering analysis. Differentially expressed genes between different CRG clusters were identified. The differentially expressed genes were then used to build a risk assessment model using least absolute shrinkage and selection operator regression to predict patient survival with BRCA. The model was then validated with the data from the Molecular Taxonomy of Breast Cancer International Consortium, GSE96058, and GSE20685. Differences in somatic mutations, copy number variations, hallmark pathways, drug responses, and prognosis of immunotherapy and chemotherapy were analyzed by comparing the high-risk and low-risk groups. Patients with high-risk scores showed worse overall survival than those with low-risk scores. The results indicated significant differences between the 2 groups immune-related biological pathways and the variable immune status. It also suggests the differential sensitivity to chemotherapy between the 2 groups. The CRGs model showed the promise to predict the prognosis of BRCA patients and shed light on their treatment.
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Neoplasias da Mama , Humanos , Feminino , Neoplasias da Mama/genética , Variações do Número de Cópias de DNA , Prognóstico , Morte Celular , Análise por Conglomerados , ApoptoseRESUMO
Background: Thyroid carcinoma (THCA), the most common endocrine neoplasm, typically exhibits an indolent behavior. However, in some instances, lymph node metastasis (LNM) may occur in the early stages, with the underlying mechanisms not yet fully understood. Materials and methods: LNM potential was defined as the tumor's capability to metastasize to lymph nodes at an early stage, even when the tumor volume is small. We performed differential expression analysis using the 'Limma' R package and conducted enrichment analyses using the Metascape tool. Co-expression networks were established using the 'WGCNA' R package, with the soft threshold power determined by the 'pickSoftThreshold' algorithm. For unsupervised clustering, we utilized the 'ConsensusCluster Plus' R package. To determine the topological features and degree centralities of each node (protein) within the Protein-Protein Interaction (PPI) network, we used the CytoNCA plugin integrated with the Cytoscape tool. Immune cell infiltration was assessed using the Immune Cell Abundance Identifier (ImmuCellAI) database. We applied the Least Absolute Shrinkage and Selection Operator (LASSO), Support Vector Machine (SVM), and Random Forest (RF) algorithms individually, with the 'glmnet,' 'e1071,' and 'randomForest' R packages, respectively. Ridge regression was performed using the 'oncoPredict' algorithm, and all the predictions were based on data from the Genomics of Drug Sensitivity in Cancer (GDSC) database. To ascertain the protein expression levels and subcellular localization of genes, we consulted the Human Protein Atlas (HPA) database. Molecular docking was carried out using the mcule 1-click Docking server online. Experimental validation of gene and protein expression levels was conducted through Real-Time Quantitative PCR (RT-qPCR) and immunohistochemistry (IHC) assays. Results: Through WGCNA and PPI network analysis, we identified twelve hub genes as the most relevant to LNM potential from these two modules. These 12 hub genes displayed differential expression in THCA and exhibited significant correlations with the downregulation of neutrophil infiltration, as well as the upregulation of dendritic cell and macrophage infiltration, along with activation of the EMT pathway in THCA. We propose a novel molecular classification approach and provide an online web-based nomogram for evaluating the LNM potential of THCA (http://www.empowerstats.net/pmodel/?m=17617_LNM). Machine learning algorithms have identified ERBB3 as the most critical gene associated with LNM potential in THCA. ERBB3 exhibits high expression in patients with THCA who have experienced LNM or have advanced-stage disease. The differential methylation levels partially explain this differential expression of ERBB3. ROC analysis has identified ERBB3 as a diagnostic marker for THCA (AUC=0.89), THCA with high LNM potential (AUC=0.75), and lymph nodes with tumor metastasis (AUC=0.86). We have presented a comprehensive review of endocrine disruptor chemical (EDC) exposures, environmental toxins, and pharmacological agents that may potentially impact LNM potential. Molecular docking revealed a docking score of -10.1 kcal/mol for Lapatinib and ERBB3, indicating a strong binding affinity. Conclusion: In conclusion, our study, utilizing bioinformatics analysis techniques, identified gene modules and hub genes influencing LNM potential in THCA patients. ERBB3 was identified as a key gene with therapeutic implications. We have also developed a novel molecular classification approach and a user-friendly web-based nomogram tool for assessing LNM potential. These findings pave the way for investigations into the mechanisms underlying differences in LNM potential and provide guidance for personalized clinical treatment plans.
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Neoplasias da Glândula Tireoide , Humanos , Algoritmos , Metástase Linfática , Simulação de Acoplamento Molecular , Mapas de Interação de Proteínas , Neoplasias da Glândula Tireoide/genéticaRESUMO
BACKGROUND: Homologous repair deficiency (HRD) causes double-strand break repair to be impeded, which is a common driver of carcinogenesis. However, the therapeutic and prognostic potential of HRD in invasive breast cancer (BRCA) has not been fully explored using comprehensive bioinformatics analysis. MATERIALS AND METHODS: HRD score was defined as the unweighted sum of LOH, TAI, and LST scores and obtained from the previous study according to Theo A et al. To characterize BRCA tumor microenvironment (TME) subtypes, "ConsensusClusterPlus" R package was used to conduct unsupervised clustering. The xCell algorithm was utilized for tumor composition analysis to estimate the TME in TCGA-BRCA. A WGCNA analysis was conducted to uncover the gene coexpression modules and hub genes in the HRD-related gene module of BRCA. The functional enrichment study was carried out using Metascape. A novel analysis pipeline, Genetic Perturbation Similarity Analysis (GPSA), was used to explore the single-gene perturbation closely related to HRD based on 3048 stable knockdown/knockout cell lines. The prognostic variables were evaluated using univariate COX analysis. Kaplan-Meier (KM) survival analysis was performed to assess the prognostic potential of HRD score. Receiver operator characteristic (ROC) curve was utilized to judge the diagnostic utility. Drug sensitivity was estimated through the R package "oncoPredict" and Genomics of Drug Sensitivity in Cancer (GDSC) database. XSum algorithm was performed to screen the candidate small molecule drugs based on the connectivity map (CMAP) database. RESULTS: Low HRD score suggested a better prognosis in BRCA patients. The tumor with low HRD score had considerably greater degree of infiltration of stromal cells and infiltration of immunocytes was significantly enhanced in the high HRD score group. Using WGCNA, ten co-expression modules were obtained. The turquoise module and 25 hub genes were identified as the most correlated with HRD in BRCA. Functional enrichment analysis revealed that the turquoise gene module was mainly concentrated in the "cell cycle" pathways. Candidate HRD-related gene signatures (MELK) were screened out through WGCNA and GPSA analysis pipeline and then validated on independent validation sets. A small molecule drug (Clofibrate) that has the potential to reverse the increase of high HRD score was screened out to improve oncological outcomes in BRCA. Molecular docking suggested MELK to be one of possible molecular targets in the Clofibrate treatment of BRCA. CONCLUSION: Based on bioinformatic analysis, we fully explored the therapeutic and prognostic potential of HRD in BRCA. A novel HRD-related gene signature (MELK) were identified through the combination of WGCNA and GPSA analysis. In addition, we detailed the TME landscape in BRCA and identified four unique TME subtypes in group with high or low HRD score group. Moreover, Clofibrate were screened out to improve oncological outcomes in BRCA by reversing the increase of high HRD score. Thus, our study contributes to the development of personalized clinical management and treatment regimens of BRCA.
Assuntos
Neoplasias da Mama , Humanos , Feminino , Clofibrato , Simulação de Acoplamento Molecular , Mama , Microambiente Tumoral , Proteínas Serina-Treonina QuinasesRESUMO
Oxidative stress is a key pathogenic factor of cerebral ischemia/reperfusion (I/R) injury. MicroRNA-32-3p (miR-32-3p) plays critical roles in regulating ischemic diseases; however, its role in oxidative stress and cerebral I/R injury remains elusive. Primary cortical neurons and rats were treated with the agomir, antagomir and matched controls of miR-32-3p, and then received oxygen glucose deprivation/reperfusion (OGD/R) or I/R stimulation. To investigate the involvement of AMP-activated protein kinase (AMPK) and calcium-binding protein 39 (Cab39), a pharmacological inhibitor and small interfering RNA were used in vivo and in vitro. Herein, we found that miR-32-3p was upregulated in OGD/R-treated neurons and I/R-injured brains, and that inhibiting miR-32-3p by the miR-32-3p antagomir dramatically alleviated oxidative stress and neural death in OGD/R-stimulated primary cortical neurons. Conversely, overexpressing miR-32-3p by the miR-32-3p agomir further aggravated OGD/R-induced neural death and oxidative damage in primary cortical neurons. Meanwhile, we observed that the miR-32-3p antagomir prevented, while the miR-32-3p agomir facilitated neural death, oxidative damage and cerebral I/R injury in vivo. Mechanistically, miR-32-3p bound to the 3'-untranslated regions of Cab39, inhibited its protein level and subsequently inactivated AMPK. Conversely, treatment with the miR-32-3p antagomir upregulated Cab39 and activated AMPK, thereby attenuating oxidative damage and cerebral I/R injury. Moreover, inhibiting AMPK or Cab39 dramatically blocked the miR-32-3p antagomir-mediated beneficial effects against cerebral I/R injury in vivo and in vitro. miR-32-3p plays critical roles in neural death and oxidative damage upon I/R stimulation, and it is a novel target to treat cerebral I/R injury.
Assuntos
Isquemia Encefálica , MicroRNAs , Traumatismo por Reperfusão , Ratos , Animais , Proteínas Quinases Ativadas por AMP , MicroRNAs/genética , Antagomirs , Traumatismo por Reperfusão/tratamento farmacológico , Isquemia Encefálica/metabolismo , Glucose/metabolismo , ApoptoseRESUMO
Objective: This study aims to evaluate the intervention effect of concurrent training on children with malignant tumors to provide evidence for prescribing exercise for children with malignant tumors. Methods: Twelve databases were searched from inception to October 15, 2022. Two researchers independently screened the literature, evaluated the quality, extracted the data, and performed the meta-analysis using R. Result: A total of nine randomized controlled trials involving 371 children were included in this study. The meta-analysis revealed that muscle strength was significantly greater in the exercise group compared to the usual care group [SMD = 0.26, 95% CI (0.04, 0.48), P = 0.023], with subgroup analysis showing no significant difference in upper limb [SMD = 0.13, 95% CI (-0.17, 0.43), P = 0.318] and a considerable difference in lower limb strength [SMD = 0.41, 95% CI (0.08, 0.74), P = 0.015]. Physical activity [SMD = 0.57, 95% CI (0.03, 1.1), P = 0.038], timed up and down stairs test [SMD = -1.22, 95% CI (-2.04, -0.4), P = 0.004], 6-min walking ability [SMD = 0.75, 95% CI (0.38, 1.11), P < 0.01], quality of life [SMD = 0.28, 95% CI (0.02, 0.53), P = 0.033], and cancer-related fatigue [SMD = -0.53, 95% CI (-0.86, -0.19), P = 0.002] were significantly better than the usual care group. There were no significant differences in peak oxygen uptake [SMD = 0.13, 95% CI (-0.18, 0.44), P = 0.397], depression [SMD = 0.06, 95% CI (-0.38, 0.5), P = 0.791], and withdrawal rates [RR = 0.59, 95% CI (0.21, 1.63), P = 0.308] between the two groups. Conclusion: Concurrent training could improve physical performance for children with malignancy but had no significant effect on mental health. Because the quality level of evidence is mostly very low, future high-quality randomized controlled trials are required to confirm these findings. Systematic review registration: https://www.crd.york.ac.uk/PROSPERO/display_record.php?RecordID=364140, identifier CRD42022308176.
Assuntos
Neoplasias , Yoga , Humanos , Criança , Qualidade de Vida , Exercício Físico , Neoplasias/terapia , Yoga/psicologia , Desempenho Físico Funcional , Ensaios Clínicos Controlados Aleatórios como AssuntoRESUMO
Bone marrow-derived mesenchymal stem cells (BMSCs) transplantation is a promising therapeutic strategy for cerebral ischemia/reperfusion (I/R) injury; however, the clinical outcome is barely satisfactory and demands further improvement. The present study aimed to investigate whether preconditioning of BMSCs by recombinant human growth differentiation factor 7 (rhGDF7) could enhance its therapeutic capacity against cerebral I/R injury. Mouse BMSCs and primary neurons were co-cultured and exposed to oxygen glucose deprivation/reperfusion (OGD/R) stimulation. To investigate the role of exosomal microRNA-369-3p (miR-369-3p), inhibitors, RNAi and the miR-369-3p antagomir were used. Meanwhile, mice were intravenously injected with rhGDF7-preconditioned BMSCs and then received cerebral I/R surgery. Markers of inflammation, oxidative stress and neural damage were evaluated. To inhibit AMP-activated protein kinase (AMPK), compound C was used in vivo and in vitro. Compared with cell-free transwell or vehicle-preconditioned BMSCs, rhGDF7-preconditioned BMSCs significantly prevented OGD/R-induced inflammation, oxidative stress and neural damage in vitro. Meanwhile, rhGDF7-preconditioned BMSCs could prevent I/R-induced cerebral inflammation and oxidative stress in vivo. Mechanistically, rhGDF7 preconditioning significantly increased exosomal miR-369-3p expression in BMSCs and then transferred exosomal miR-369-3p to primary neurons, where it bound to phosphodiesterase 4 D (Pde4d) 3'-UTR and downregulated PDE4D expression, thereby preventing I/R-induced inflammation, oxidative stress and neural damage through activating AMPK pathway. Our study identify GDF7 pretreatment as a promising adjuvant reagent to improve the therapeutic potency of BMSCs for cerebral I/R injury and ischemic stroke.