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Objective: To detect the incidence and analyze the clinical significance of regions of homozygosity (ROH) through the single nucleotide polymorphism array (SNP array). Methods: The SNP array detection results of 5 116 pregnant women in the Third Affiliated Hospital of Guangzhou Medical University from January 2016 to December 2020 were retrospectively analyzed. The pregnant women with ROH (5 Mb as the threshold) were followed up to analyze the relationship between ROH and abnormal fetal phenotype. Whole exon sequencing was performed in 4 cases of consanguineous marriage to detect potential recessive causative genes in the ROH region. Results: (1) A total of 39 cases of ROH were detected, with a positive rate of 0.76% (39/5 116). Among them, 25 cases (64%, 25/39) were detected only on single chromosome, and chromosome 11 had the highest detection rate, suggesting the risk of uniparental disomy; fourteen cases (36%,14/39) were detected on multiple chromosomes, most commonly on chromosomes 11, 1, 3, 4 and 8. (2) The number of cases and detection rate of ROH detected by different prenatal diagnosis indicators were as follows: 12 cases (1.78%, 12/676) in pregnant women with abnormal non-invasive prenatal testing result, 12 cases (0.37%, 12/3 284) in pregnant women with ultrasound abnormality, 4 cases (4/4) in pregnant women with consanguineous marriage, 3 cases (0.92%, 3/326) in pregnant women with previous adverse pregnancy, 2 cases (1.15%, 2/174) in pregnant women with high risk of serology in screening, 2 cases (4.00%, 2/50) in pregnant women with abnormal fetal chromosomal karyotype, 2 cases (0.79%, 2/253) in pregnant women with advanced maternal age, 1 case (0.56%, 1/178) in pregnant women with related parental genetic factors and 1 case (0.58%, 1/171) in pregnant women with the other factors. (3) The follow-up results of 39 cases of prenatal ROH showed that there were 16 cases of term birth, 15 cases of termination of pregnancy, 2 cases of preterm births, 1 case of fetal death and 5 cases lost to follow-up. Conclusions: Chromosomal ROH phenomenon is not rare. By analyzing the detection rate of ROH in prenatal diagnosis, combined with the results of fetal phenotype and postpartum follow-up, the clinical characteristics of ROH are discussed, so as to better understand the relationship between ROH and its phenotype.
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Cariótipo Anormal , Diagnóstico Pré-Natal , Feminino , Feto , Humanos , Gravidez , Cuidado Pré-Natal , Diagnóstico Pré-Natal/métodos , Estudos RetrospectivosRESUMO
AIMS: This study aimed to explore potential drug targets of Streptococcus suis at the system level. METHODS AND RESULTS: A homologous protein mapping method was used in the construction of a protein-protein interaction (PPI) network of S. suis, which presented 1147 non-redundant interaction pairs among 286 proteins. The parameters of PPI networks were calculated and showed scale-free network properties. In all, 41 possibly essential proteins identified from 47 highly connected proteins were selected as potential drug target candidates. Of these proteins, 30 were already regarded as drug targets in other bacterial species. Six transporters with high connections to other functional proteins were identified as probably not essential but important functional proteins. Afterward, the subnetwork centred with cell division protein FtsZ was used in confirming the PPI network through bacterial two-hybrid analysis. CONCLUSIONS: The predicted PPI network covers 13·04% of the proteome in S. suis. The selected 41 potential drug target candidates are conserved between S. suis and several model bacteria. SIGNIFICANCE AND IMPACT OF THE STUDY: The predictions included proteins known to be drug targets, and a verifying experiment confirmed the reliability of predicted interactions. This work is the first to present systematic computational PPI data for S. suis and provides potential drug targets, which are valuable in exploring novel anti-streptococcus drugs.
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Proteínas de Bactérias/metabolismo , Mapeamento de Interação de Proteínas , Streptococcus suis/metabolismo , Antibacterianos/farmacologia , Proteínas de Bactérias/antagonistas & inibidores , Proteínas de Membrana Transportadoras/metabolismo , Proteoma/metabolismo , Streptococcus suis/efeitos dos fármacosRESUMO
Objective: To investigate the effect of triptolide on radiosensitivity of lung cancer cells and its mechanism. Methods: The lung cancer cells H1299, A549, H157 and H838 were cultured. The strongest radio resistance cell line, H1299 was selected by cell clone formation experiment and for the subsequent experiments. 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H tetrazolium bromide (MTT) was used to detect the effect of different concentrations of triptolide on the proliferation of H1299 cells. The optimal concentration of triptolide was 50nmol/L, and the optimal treatment time was 48 hours. The H1299 cells were divided into the control group, triptolide group (50 nmol/L), 4 Gy group and triptolide+ 4 Gy group. Flow cytometry was used to detect the apoptosis rate of H1299 cell. Western Blot was used to detect the proteins expression levels of p-Chk2, p-ATM, p-p53, Bcl-2, Bax and cleaved-Caspase 3. Results: The apoptotic rate and protein levels of Bax, cleaved-Caspase 3, p-Chk2, p-ATM and p-p53 in the 4 Gy group were (12.38±1.34)%, 0.42±0.04, 0.38±0.04, 0.98±0.11, 0.73±0.08, 0.95±0.09, respectively, higher than (3.26±2.43)%, 0.22±0.02, 0.23±0.03, 0.32±0.03, 0.21±0.02, 0.25±0.03 in the control group (P<0.05). However, the protein level of Bcl-2 was (0.52±0.05), lower than (0.93±0.09) of the control group (P<0.05). The survival fraction (0.462) and protein level of Bcl-2 (0.44±0.04) in the triptolide group were lower than those of the control group (0.702 and 0.93±0.09, P<0.05). The apoptotic rate and the protein levels of Bax and cleaved-Caspase 3 in the triptolide group were (9.27±1.08)%, 0.45±0.05, 0.41±0.04, respectively, higher than (3.26±2.43)%, 0.22±0.02, 0.23±0.03 in the control group (P<0.05), and the sensitization ratio in the triptolide group was 1.579. The apoptosis rate, Bax and cleaved Caspase 3 protein expression levels in triptolide + 4 Gy group were (26.53±2.19)%, 0.91±0.09 and 0.79±0.08, respectively, higher than (12.38±1.34)%, 0.42±0.04 and 0.38±0.04 in 4 Gy group (P<0.05). The expression level of Bcl-2 protein was (0.21±0.02), lower than (0.52±0.05) in 4 Gy group (P<0.05). Conclusion: Triptolide increases the radiosensitivity of radiation-induced lung cancer cells by inhibiting DNA repair and inducing apoptosis.
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Neoplasias Pulmonares , Tolerância a Radiação , Apoptose , Reparo do DNA , Diterpenos , Compostos de Epóxi , Humanos , FenantrenosRESUMO
ABSTRACT: Objective To study the changes of metabolites in serum and tissues ï¼kidney, liver and heartï¼ of mice died of acute tetracaine poisoning by metabolomics, to search for potential biomarkers and related metabolic pathways, and to provide new ideas for the identification of cause of death and research on toxicological mechanism of acute tetracaine poisoning. Methods Forty ICR mice were randomly divided into control group and acute tetracaine poisoning death group. The model of death from acute poisoning was established by intraperitoneal injection of tetracaine, and the metabolic profile of serum and tissues of mice was obtained by ultra-high performance liquid chromatography-electrostatic field orbitrap high resolution mass spectrometry ï¼UPLC-Orbitrap HRMSï¼. Multivariate statistical principal component analysis ï¼PCAï¼ and orthogonal partial least square-discriminant analysis ï¼OPLS-DAï¼ were used, combined with t-test and fold change to identify the differential metabolites associated with death from acute tetracaine poisoning. Results Compared with the control group, the metabolic profiles of serum and tissues in the mice from acute tetracaine poisoning death group were significantly different. Eleven differential metabolites were identified in serum, including xanthine, spermine, 3-hydroxybutylamine, etc.; twenty-five differential metabolites were identified in liver, including adenylate, adenosine, citric acid, etc.; twelve differential metabolites were identified in heart, including hypoxanthine, guanine, guanosine, etc; four differential metabolites were identified in kidney, including taurochenodeoxycholic acid, 11, 12-epoxyeicosatrienoic acid, dimethylethanolamine and indole. Acute tetracaine poisoning mainly affected purine metabolism, tricarboxylic acid cycle, as well as metabolism of alanine, aspartic acid and glutamic acid. Conclusion The differential metabolites in serum and tissues of mice died of acute tetracaine poisoning are expected to be candidate biomarkers for this cause of death. The results can provide research basis for the mechanism and identification of acute tetracaine poisoning.
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Metabolômica , Tetracaína , Animais , Biomarcadores/metabolismo , Cromatografia Líquida de Alta Pressão , Espectrometria de Massas , Metaboloma , Camundongos , Camundongos Endogâmicos ICRRESUMO
Heavy metal pollution from industrial wastewater is an important source. A method for heavy metals determination in industrial wastewater based on laser-induced breakdown spectroscopy (LIBS) technique was studied and the on-line monitoring system that used automatic graphite enrichment and spatial plasma confinement detection was developed and field demonstrated. The limits of detection (LOD) of heavy metal elements (Cd, Cr, Cu, Ni, Pb, Zn) could reach several µg/L. In Tongling, the on-line heavy metal monitor was field demonstrated. The calibration curves of copper and zinc were built on site, and then on-line monitoring was conducted. The measurement results of this monitor were compared with ICP-OES and had a good correlation. The results showed that the heavy metal monitor could be used for on-line detection of heavy metals in wastewater and had a good reliability.
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OBJECTIVES: To compare adverse effects, tolerability and efficacy of the tricyclic antidepressants (TCAs) amitriptyline and nortriptyline in management of neuropathic pain due to peripheral neuropathy (PN). MATERIALS & METHODS: We performed a prospective open-label flexible-dosing comparison of monotherapy or adjuvant therapy using amitriptyline or nortriptyline in PN-associated neuropathic pain. Primary outcomes were quantitative adverse effects and discontinuation rates. Secondary outcomes assessed changes in pain severity, quality of life, disability, sleep efficacy, mood and anxiety, and global improvement. Assessments occurred at 3 and 6 months after initiation. Our hypothesis was that nortriptyline would have better tolerance than amitriptyline. RESULTS: A total of 228 PN patients were enrolled approximately equally for monotherapy and adjuvant therapy. Adverse effects and discontinuation rates were similar between amitriptyline and nortriptyline interventions. Weight gain was more common with amitriptyline, while nortriptyline use was associated with greater prevalence of dry mouth. Secondary outcome measures were similar in both groups, demonstrating improvement from baseline. CONCLUSIONS: Amitriptyline and nortriptyline are equivalent for overall adverse effects and discontinuation rates. Either TCA should be equally considered for use in neuropathic pain due to PN. When used as monotherapy or as part of adjuvant therapy, either TCA can be expected to provide approximately 23-26% visual analog scale pain reduction if tolerated. Discontinuations due to inefficacy or adverse effects can be anticipated in 26-37% of patients initiated on either TCA for PN-associated neuropathic pain.
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Amitriptilina/uso terapêutico , Antidepressivos Tricíclicos/uso terapêutico , Neuralgia/tratamento farmacológico , Nortriptilina/uso terapêutico , Doenças do Sistema Nervoso Periférico/tratamento farmacológico , Amitriptilina/efeitos adversos , Antidepressivos Tricíclicos/efeitos adversos , Quimioterapia Combinada , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Neuralgia/etiologia , Nortriptilina/efeitos adversos , Medição da Dor , Doenças do Sistema Nervoso Periférico/complicações , Equivalência Terapêutica , Resultado do Tratamento , Recusa do Paciente ao TratamentoRESUMO
The magnetic properties of a Co2FeAl/(Ga,Mn)As bilayer epitaxied on GaAs (001) are studied both experimentally and theoretically. Unlike the common antiferromagnetic interfacial interaction existing in most ferromagnet-magnetic semiconductor bilayers, a ferromagnetic interfacial interaction in the Co2FeAl/(Ga,Mn)As bilayer is observed from measurements of magnetic hysteresis and x-ray magnetic circular dichroism. The Mn ions in a 1.36 nm thick (Ga,Mn)As layer remain spin polarized up to 400 K due to the magnetic proximity effect. The minor loops of the Co2FeAl/(Ga,Mn)As bilayer shift with a small ferromagnetic interaction field of +24 Oe and -23 Oe at 15 K. The observed ferromagnetic interfacial coupling is supported by ab initio density functional calculations. These findings may provide a viable pathway for designing room-temperature semiconductor spintronic devices through magnetic proximity effect.
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OBJECTIVE: Adult stem cell senescence and exhaustion are important drivers of organismal age. Restored stem cell self-renewal has revealed novel therapeutic targets for decreasing the incidence of age-associated diseases (AADs) and prolonging the human health span. Transient ectopic expression of the reprogramming factors Oct3/4, Sox2, Klf4 and c-Myc (collectively known as OSKM) in somatic cells can induce partial cellular reprogramming and effectively ameliorate their age-associated hallmarks. However, how this form of rejuvenation is applied to senescent stem cells remains unknown. MATERIALS AND METHODS: The Integrin-α6highCD71high epidermal stem cells (ESCs) with low self-renewal ability were sorted by flow cytometry and then treated by the interrupted reprogramming induced by transient expression of OSKM. The ability of secondary clones' generation and self-proliferation in vitro, as well as stem cell marker p63, were detected to determine their self-renewal ability. Besides, gene and protein of epidermal cell markers were detected to determine whether their cell identities were retained. Finally, DNA methylation age (eAge) and DNA dehydroxymethylase/methyltransferase were analyzed to explore the alternation of their global DNA methylation pattern during this rejuvenation. RESULTS: The partial reprogramming restored the youthful self-renewal and proliferation in senescent ESCs, including larger secondary clone generation, higher expression of stem cell marker p63 and proliferation marker Ki67, and faster proliferation speed, in each case without abolishing epithelial cellular identity. Moreover, the rejuvenation of adult stem cells could be maintained for 2 weeks after reprogramming factor withdrawal, which was more stable than that of differentiated somatic cells. Additionally, we found that partial reprogramming counteracted the acceleration of eAge in senescent epidermal stem cells and DNA methyltransferase 1 (DNMT1) may play a crucial role in this process. CONCLUSIONS: Partial reprogramming has high therapeutic potential for reversing adult stem cell age, providing an advanced way to treat AADs.
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Reprogramação Celular , Células-Tronco Pluripotentes Induzidas , Adulto , Humanos , Células-Tronco , Células Epidérmicas , Metiltransferases/metabolismo , DNA/metabolismo , Células-Tronco Pluripotentes Induzidas/metabolismoRESUMO
Objective: To analyze the difference between Z score and previous criteria in the diagnosis characteristics of coronary artery aneurysm (CAA) in Kawasaki disease, and to investigate the clinical distribution of Kawasaki disease CAA in the Z score group. Methods: This study retrospectively analyzed the clinical and echocardiographic data of 2 419 children with Kawasaki disease in Shenzhen Children's Hospital from January 2009 to December 2019. The traditional criteria and Z score criteria were used to diagnose CAA, and the differences of diagnostic efficiency between the 2 diagnostic methods were analyzed. The clinical distribution characteristics of CAA in children with Kawasaki disease were analyzed by grouping their sex, clinical classification (complete Kawasaki disease, incomplete Kawasaki disease) the sensitivity to intravenous immunoglobulin (IVIG) (IVIG-sensitive Kawasaki disease,IVIG-unresponsive Kawasaki disease). And the course of the disease (≤6 weeks, >6-8 weeks, >8 weeks to 6 months) etc. The χ² test or Kruskal-Wallis test was used for comparison between the groups, and the Kappa test was used for consistency evaluation. Results: Among the 2 419 children with Kawasaki disease, 1 558 were males and 861 were females. The age of onset was 1.8 (1.0, 3.2) years. The rate of CAA by Z score criteria was higher than that by traditional method (21.9% (529/2 419) vs. 13.9% (336/2 419), χ2=1 074.94, P<0.001). Compared to the traditional method, the Z score criteria found higher rate of CAA in male patients, patients with incomplete Kawasaki disease, and IVIG-unresponsive patients (25.2% (392/1 558) vs. 16.0% (249/1 558), (32.7% (166/507) vs. 19.5% (99/507), 30.5% (95/312) vs. 24.0% (75/312), χ2=694.05, 216.19, 184.37, all P<0.001). The Z score criteria was consistent with the traditional method in diagnosing CAA (κ=0.642,P<0.001). Moreover, in the Z score criteria, the rate of CAA in males (25.2%, 392/1 558) was higher than that in females (15.9%, 137/861), higher in incomplete Kawasaki cases (32.7%, 166/507) than that in complete Kawasaki case (19.0%, 363/1 912), and higher in IVIG-unresponsive cases (30.4%, 95/312) than that in IVIG-sensitive cases (20.6%, 434/2 107), with statistically significant differences (χ2=27.76, 44.38, 15.43, all P<0.001). Coronary Z score of course ≤ 6 weeks was greater than that of course between>6-8 weeks and >8 weeks to 6 months (1.3 (0.7, 2.3) vs. 0.7 (0.3, 1.4), 0.7 (0.3, 1.3), Z=20.65, 13.70, both P<0.001). Conclusions: The rate of CAA in Kawasaki disease by Z score criteria is higher than that by traditional method. In the Z score group, most CAA occur within 6 weeks of the course of the disease, and the rate of CAA in male, incomplete Kawasaki disease, and IVIG-unresponsive is higher.
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Aneurisma Coronário , Doença da Artéria Coronariana , Síndrome de Linfonodos Mucocutâneos , Criança , Aneurisma Coronário/diagnóstico por imagem , Aneurisma Coronário/etiologia , Vasos Coronários/diagnóstico por imagem , Ecocardiografia , Feminino , Humanos , Imunoglobulinas Intravenosas/uso terapêutico , Lactente , Masculino , Síndrome de Linfonodos Mucocutâneos/complicações , Síndrome de Linfonodos Mucocutâneos/diagnóstico por imagem , Estudos RetrospectivosRESUMO
The frequent disease outbreaks caused by avian influenza virus not only affect the poultry industry but also pose a threat to human safety. To address the problem, RNA interference (RNAi) has recently been widely used as a potential antiviral approach. Transgenesis in combination with RNAi to specifically inhibit avian enza virus gene expression has been proposed to make chickens resistant to the infection. For the transgenic breeding, screening in vitro efficient siRNAs as the candidate genes is one of the most important tasks. Here, we combined an online search tool and a series of bioinformatics programs with a set of rules for designing siRNAs targeted towards different mRNA regions of H5N1 avian influenza virus. Five rational siRNAs were chosen by this method, five U6 promoter-driven shRNA expression plasmids containing the siRNA genes were constructed and used for producing stably transfected MDCK cells. The data obtained by virus titration, IFA, PI-stained flow cytometry, real-time quantitative RT-PCR, and DAS-ELISA analyses showed that all five stably transfected cell lines we re resistant to virusreplication when exposed to 100 CCID50 of avian influenza virus H5N1. Finally, most effective plasmids (pSi-604i and pSi-1597i) as the candidates for making the transgenic chickens were chosen. These findings provide baseline information on use of RNAi technique for breeding transgenic chickens resistant to avian influenza virus.
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Animais Geneticamente Modificados/genética , Cruzamento , Galinhas/genética , Virus da Influenza A Subtipo H5N1 , Influenza Aviária/genética , RNA Interferente Pequeno/genética , Animais , Animais Geneticamente Modificados/virologia , Sequência de Bases , Linhagem Celular Transformada , Galinhas/virologia , Cães , Conformação de Ácido Nucleico , Plasmídeos/genética , RNA Interferente Pequeno/isolamento & purificaçãoRESUMO
The article "Preclinical pharmacokinetic study of a novel lipid-lowering agent, IMM-H007, by Z.-L. Zhang, W.-Q. Liu, X.-Z. Deng, published in Eur Rev Med Pharmacol Sci 2018; 22 (24): 8939-8950-DOI: 10.26355/eurrev_201812_16664-PMID: 30575938" has been withdrawn from the authors stating that "after our article was published, we received a conflict of interest from the Chinese Academy of Sciences. They declared that they developed IMM-H007 for the first time and they had already applied for the patent. We had already cited in the references that IMM-H007 came from the Chinese Academy of Sciences. But they still insisted that we had violated their rights. We had to withdraw our paper in order to avoid greater conflicts and disputes". The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/16664.
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Several bioactive molecules isolated from the saliva of blood-sucking arthropods, such as mosquitoes, have been shown to exhibit potential anticoagulant function. We have previously identified a 30kDa allergen named Aegyptin-like protein (alALP), which is highly homologous to Aegyptin, from the salivary glands of female Aedes albopictus (Asian tiger mosquito). In this study, we identified the conserved functional domain of alALP by using bioinformatic tools, and expressed the His-tagged alALP recombinant protein in sf9 insect cells by generation and transfection of a baculoviral expression plasmid carrying the fulllength cDNA of alALP. We purified this recombinant protein and examined its function on the inhibition of blood coagulation. The results showed that the purified His-alALP prolonged the Activated Partial Thromboplastin Time (APTT), Prothrombin Time (PT) and Thrombin Time (TT) in vitro as well as the Bleeding Time (BT) in vivo, which suggest that alALP could be a novel anticoagulant.
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Aedes/genética , Anticoagulantes/química , Proteínas de Insetos/química , Proteínas e Peptídeos Salivares/química , Sequência de Aminoácidos , Animais , Testes de Coagulação Sanguínea , Clonagem Molecular , Biologia Computacional , Proteínas de Insetos/genética , Camundongos , Tempo de Tromboplastina Parcial , Tempo de Protrombina , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas e Peptídeos Salivares/genéticaRESUMO
We identify an essential role for the RGD (Arg-Gly-Asp tripeptide) moiety in vivo during adult peripheral neuron regenerative growth. Beyond a peripheral nerve transection there were rises in the fibronectin extracellular matrix, and striking rises in the mRNA and protein expression of integrin subunits sensitive to RGD/fibronectin signalling. Neuron perikarya, axons and Schwann cells all expressed RGD/fibronectin sensitive integrins after injury. To evaluate the significance of RGD/fibronectin-integrin interactions, we infused solutions of a pentapeptide including the RGD motif (sRGD) serially and directly within the milieu of early axon growth across rat sciatic transection injuries. While low dose infusions of sRGD facilitated early axon ingrowth, we encountered inhibition of ingrowth and bridge formation with higher doses of sRGD indicating competitive disruption of RGD/fibronectin-integrin signalling. Fibronectin RGD moieties serve a critical and important role during peripheral axon outgrowth.
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Axônios/fisiologia , Regeneração Nervosa/fisiologia , Oligopeptídeos/metabolismo , Nervo Isquiático/fisiologia , Análise de Variância , Animais , Matriz Extracelular/metabolismo , Fibronectinas/metabolismo , Imuno-Histoquímica , Integrinas/metabolismo , Masculino , Fibras Nervosas Mielinizadas/fisiologia , Oligopeptídeos/farmacologia , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Células de Schwann/metabolismo , Nervo Isquiático/lesões , Transdução de SinaisRESUMO
OBJECTIVE: The aim of this study was to investigate the effect of long non-coding ribonucleic acid (lncRNA) nuclear paraspeckle assembly transcript 1 (NEAT1) on sepsis-induced brain injury in mice through nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB). MATERIALS AND METHODS: The mouse model of sepsis was established by cecal ligation and puncture induction. The relative expression levels of NEAT1 and NF-κB in brain tissues of mice in healthy group and sepsis group were determined via quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR) and Western blotting, respectively. Subsequently, the expression of NEAT1 was silenced by transfection of small interfering RNAs (siRNAs). Meanwhile, its effect on NF-κB expression was detected. To further explore the effect of sepsis on brain injury, the content of brain water and the expression levels of apoptosis-related proteins, including B-cell lymphoma 2 (Bcl-2) and Bcl-2-associated X protein (BAX), in mice of healthy group, sepsis group, and sepsis + si-NEAT1 group were measured. Furthermore, MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assays were used to detect the proliferation and apoptosis of nerve cells. RESULTS: The relative expression levels of NEAT1 and NF-κB were significantly increased in the brain tissues of septic mice (p<0.01). Si-NEAT1 transfection significantly decreased the expressions of NEAT1 and NF-κB in brain tissues of septic mice (p<0.05). The content of brain water in mice of sepsis group was evidently increased (p<0.05). However, si-NEAT1 treatment remarkably reduced this content (p<0.05). In addition, sepsis markedly decreased the activity of nerve cells (p<0.05). However, si-NEAT1 could significantly increase the activity of nerve cells in septic mice (p<0.05). Moreover, si-NEAT1 notably decreased the expression of BAX (p<0.05), whereas it increased the expression of Bcl-2 (p<0.05). The results of apoptosis detection revealed that sepsis remarkably promoted the apoptosis of mouse nerve cells (p<0.05). In addition, si-NEAT1 transfection could evidently alleviate the apoptosis of nerve cells in septic mice (p<0.05). CONCLUSIONS: LncRNA NEAT1 promotes brain injury in septic mice by positively regulating NF-κB. However, si-NEAT1 transfection can reduce this injury.
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Lesões Encefálicas/patologia , NF-kappa B/metabolismo , RNA Longo não Codificante/metabolismo , Sepse/patologia , Animais , Apoptose , Encéfalo/metabolismo , Lesões Encefálicas/etiologia , Modelos Animais de Doenças , Regulação para Baixo , Camundongos , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Interferência de RNA , RNA Longo não Codificante/antagonistas & inibidores , RNA Longo não Codificante/genética , RNA Interferente Pequeno/metabolismo , Sepse/complicações , Transdução de Sinais , Regulação para Cima , Proteína X Associada a bcl-2/metabolismoRESUMO
Erythropoietin (EPO) and its receptor (EPO-R), mediate neuroprotection from axonopathy and apoptosis in the peripheral nervous system (PNS). We examined the impact and potential mechanisms of local EPO signaling on regenerating PNS axons in vivo and in vitro. As a consequence of injury, peripheral nerve axons and DRG neurons have a marked increase in the expression of EPO and EPO-R. Local delivery of EPO via conduit over 2 weeks to rat sciatic nerve following crush injury increased the density and maturity of regenerating myelinated axons growing distally from the crush site. In addition, EPO also rescued retrograde degeneration and atrophy of axons. EPO substantially increased the density and intensity of calcitonin gene-related peptide (CGRP) expression within outgrowing axons. Behavioral improvements in sensorimotor function also occurred in rats exposed to near nerve EPO delivery. EPO delivery led to decreased nuclear factor kappaB (NFkB) activation but increased phosphorylation of Akt and STAT3 within nerve and dorsal root ganglia neurons indicating rescue from an injury phenotype. Spinal cord explant studies also demonstrated a similar dose-dependent effect of EPO upon motor axonal outgrowth. Local EPO signaling enhances regenerating peripheral nervous system axons in addition to its known neuroprotection. Exogenous EPO may have a therapeutic role in a large number of peripheral nerve diseases through its impact on regeneration.
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Axônios/efeitos dos fármacos , Eritropoetina/farmacologia , Regeneração Nervosa/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Animais , Comportamento Animal/efeitos dos fármacos , Western Blotting , Relação Dose-Resposta a Droga , Regulação para Baixo/efeitos dos fármacos , Eletrofisiologia , Gânglios Espinais/efeitos dos fármacos , Gânglios Espinais/patologia , Força da Mão , Imuno-Histoquímica , Masculino , Compressão Nervosa , Condução Nervosa/efeitos dos fármacos , Técnicas de Cultura de Órgãos , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Ratos , Ratos Sprague-Dawley , Proteínas Recombinantes , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Nervo Isquiático/lesões , Nervo Isquiático/patologia , Nervo Isquiático/fisiologia , Medula Espinal/fisiologiaRESUMO
Amyotrophic lateral sclerosis (ALS) is a fatal neuromuscular disorder in which motor neurons may be targeted by oxidative and nitrergic stress without sufficient compensation by intrinsic support mechanisms. In this work, we addressed two key tenets of this hypothesis for the pathogenesis of ALS. Using superoxide dismutase (SOD) 1G93A mice, we studied the impact of reduction of nitrergic stress within the CNS with the use of a broad spectrum nitric oxide synthase (NOS) inhibitor, NG-nitro-l-arginine methyl ester. A separate cohort of SOD1G93A mice received direct insulin neurotrophic support, ligating receptors expressed upon motor neurons, to attempt protection against neuronal and functional motor dropout. For direct access, we used a novel form of intranasal delivery that provides peak concentration levels in the CNS within 1 h of delivery without systemic side effects at doses which previously rescued retrograde loss of motor axons after axotomy. To identify even minor impacts of these interventions on the outcome, we utilized an intensive program of serial behavioral and electrophysiological testing weekly, combined with endpoint quantitative morphometry and molecular analysis. This intensive evaluation enhanced our knowledge of the time course in SOD1G93A mice and impact of the SOD1G93A mutation upon motor neurons and their function. Neither intervention had even minimal impact upon slowing progression of disease in SOD1G93A mice. Our data argue against significant roles for nitrergic stress in promoting motor neuron loss and the importance of alternative neurotrophic support mechanisms that might support motor neurons and prevent disease progression in SOD1G93A mice.
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Esclerose Lateral Amiotrófica/tratamento farmacológico , Inibidores Enzimáticos/administração & dosagem , Insulina/administração & dosagem , NG-Nitroarginina Metil Éster/administração & dosagem , Potenciais de Ação/efeitos dos fármacos , Potenciais de Ação/fisiologia , Administração Intranasal , Fatores Etários , Esclerose Lateral Amiotrófica/genética , Esclerose Lateral Amiotrófica/patologia , Esclerose Lateral Amiotrófica/fisiopatologia , Animais , Comportamento Animal/efeitos dos fármacos , Modelos Animais de Doenças , Regulação da Expressão Gênica/efeitos dos fármacos , Regulação da Expressão Gênica/genética , Humanos , Camundongos , Camundongos Transgênicos , Atividade Motora/efeitos dos fármacos , Neurônios Motores/efeitos dos fármacos , Neurônios Motores/patologia , Força Muscular/efeitos dos fármacos , Músculo Esquelético/efeitos dos fármacos , Músculo Esquelético/fisiopatologia , Mutação/genética , Condução Nervosa/efeitos dos fármacos , Junção Neuromuscular/efeitos dos fármacos , Junção Neuromuscular/patologia , Nervos Periféricos/patologia , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/genética , Transdução de Sinais/fisiologia , Medula Espinal/patologia , Superóxido Dismutase/genéticaRESUMO
OBJECTIVE: To investigate the pharmacokinetic characteristics of absorption, distribution, metabolism, and excretion in vivo after oral administration and sublingual venous injection of the small molecule IMM-H007 in hamsters. MATERIALS AND METHODS: Pharmacokinetic characteristics, including absorption, distribution, metabolism, and excretion, were studied in vivo by LC-MS/MS after oral administration and sublingual venous injection of IMM-H007 in hamsters. Furthermore, IMM-H007 stability in artificial gastric juices, artificial intestinal juices, and Tris-HCl buffer was also analyzed. RESULTS: There was no significant matrix or impurity interference in golden hamster whole blood as shown using MS/MS analysis to detect the existence of these substances. IMM-H007, Ml, and MP exhibited good linearity in the range of 1-500 ng/mL, 2-1000 ng/mL, and 10-5000 ng/mL, respectively. The matrix effect was 71.93-105.49%, and IMM-H007, M1, and MP were stable during the process of sample disposal. These results illustrate that the HPLC MS/MS analytic method is simple, reliable, and sensitive and exhibits high specificity and which meets the clinical pharmacokinetic requirements of IMM-H007. IMM-H007 is rapidly absorbed through the oral route in hamsters. The Cmax and AUC(0-t) of the Ml and MP metabolites in male and female hamsters were increased with increasing dosage and were proportional to the dose. In addition, T1/2 and MRT(0-t) were significantly prolonged with increasing dosage, exhibiting linear dynamic characteristics and no significant gender differences. Bioavailability in male and female golden hamsters after oral administration of IMM-H007 was calculated using the sum of Ml and MP, resulting in 6.97% and 8.95%, respectively. IMM-H007 and its metabolites were stable in Tris-HCl buffer, artificial gastric juices, and artificial intestinal juices. CONCLUSIONS: We provide an experimental basis for elucidating the material pharmacodynamics actions of IMM-H007 and predicting its potential drug interactions.
Assuntos
Adenosina/análogos & derivados , Hipolipemiantes/farmacocinética , Adenosina/administração & dosagem , Adenosina/sangue , Adenosina/farmacocinética , Administração Intravenosa , Administração Oral , Animais , Estabilidade de Medicamentos , Feminino , Absorção Gastrointestinal , Hipolipemiantes/administração & dosagem , Hipolipemiantes/sangue , Eliminação Intestinal , Modelos Lineares , Masculino , Mesocricetus , Modelos Biológicos , Eliminação RenalRESUMO
A charge amplifier or a voltage amplifier can be used as a signal conditioning circuit between a piezoelectric element and a meter or a data acquisition board. The outputs of the piezoelectric sensor are in an open-circuit state and a short-circuit state if a voltage amplifier and a charge amplifier are used, respectively. When the electrodes are in different states, the piezoelectric element has rather different stiffness and thus different sensor resonant frequency. This phenomenon is theoretically analyzed in detail and validated by a carefully designed experiment. The results indicate that a much wider range of working frequency is achieved when a voltage amplifier is used.
RESUMO
Successful spin injection into graphene makes it a competitive contender in the race to become a key material for quantum computation, or the spin-operation-based data processing and sensing. Engineering ferromagnetic metal (FM)/graphene heterojunctions is one of the most promising avenues to realise it, however, their interface magnetism remains an open question up to this day. In any proposed FM/graphene spintronic devices, the best opportunity for spin transport could only be achieved where no magnetic dead layer exists at the FM/graphene interface. Here we present a comprehensive study of the epitaxial Fe/graphene interface by means of X-ray magnetic circular dichroism (XMCD) and density functional theory (DFT) calculations. The experiment has been performed using a specially designed FM1/FM2/graphene structure that to a large extent restores the realistic case of the proposed graphene-based transistors. We have quantitatively observed a reduced but still sizable magnetic moments of the epitaxial Fe ML on graphene, which is well resembled by simulations and can be attributed to the strong hybridization between the Fe 3dz2 and the C 2pz orbitals and the sp-orbital-like behavior of the Fe 3d electrons due to the presence of graphene.