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1.
Hepatology ; 2020 Mar 10.
Artigo em Inglês | MEDLINE | ID: mdl-32155285

RESUMO

Hepatic ischemia-reperfusion (IR) injury is a major complication of liver transplantation, resection and hemorrhagic shock. Hypoxia is a key pathological event associated with IR injury. miR-210 has been characterized as a micromanager of hypoxia pathway. However, its function and mechanism in hepatic IR injury is unknown. In this study, we found miR-210 was induced in liver tissues from patients subjected to IR related surgeries. In a murine model of hepatic IR, the level of miR-210 was increased in hepatocytes but not in nonparenchymal cells. miR-210 deficiency remarkably alleviated liver injury, cell inflammatory responses and cell death in a mouse hepatic IR model. In vitro, inhibition of miR-210 decreased HR-induced cell apoptosis of primary hepatocytes and LO2 cells, whereas overexpression of miR-210 increased cells apoptosis during HR. Mechanistically, miR-210 directly suppressed SMAD4 expression under normoxia and hypoxia condition by directly binding to the 3' UTR of SMAD4. The pro-apoptotic effect of miR-210 was alleviated by SMAD4, while sh-SMAD4 abrogated the anti-apoptotic role of miR-210 inhibition in primary hepatocytes. Further studies demonstrated that hypoxia-induced SMAD4 transported into nucleus, in which SMAD4 directly bound to the promoter of miR-210 and transcriptionally induced miR-210, thus forming a negative feedback loop with miR-210. Our study implicates a crucial role of miR-210-SMAD4 interaction in hepatic IR-induced cell death and provides a promising therapeutic approach for liver IR injury.

2.
Biomed Pharmacother ; 120: 109385, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31542613

RESUMO

Long non-coding RNA (lncRNA) linc00152 has been recognized as an oncogenic lncRNA in various cancers. This study attempts to investigate the roles of linc00152 in the metastasis-related traits of infantile hemangioma (IH). Linc00152 was overexpressed in the proliferating-phase hemangioma tissues when compared with that in involuting-phase. Downregulation of linc00152 strikingly suppressed the cell viability, migration and invasion of hemangioma cells. Furthermore, silence of linc00152 repressed the growth and lung metastasis of hemangioma cell in vivo. Subsequent analysis revealed that linc00152 bound with miR-139-5p and linc00152 expression was inversely correlated with the level of miR-139-5p in IH. Same effects of miR-139-5p transfection on HemECs cells were observed as downregulation of linc00152. Moreover, tumor protein D52 (TPD52) was confirmed to be the target of miR-139-5p. Besides, the anti-tumor effect of linc00152 silence on hemangioma cell was reversed by rexpression of TPD52. This study demonstrates that downregulatuon of linc00152 restrains the aggressiveness of hemangioma cell in vitro and in vivo via interacting with miR-139-5p and further modulates the level of TPD52.

3.
Neuropsychiatr Dis Treat ; 15: 1403-1411, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31190842

RESUMO

Background: Poststroke depression (PSD) is the most frequent psychological sequela after stroke. Electroacupuncture (EA) treatment is effective for PSD. The study aimed at clarifying the mechanisms of EA's antidepressant effects in a PSD rat model. Methods: We used middle cerebral artery occlusion to establish the rat model of PSD. Tests of sucrose preference and locomotor activity were performed to examine depressive-like behaviors. We measured malondialdehyde, GSH, SOD, IL6, IL1ß, TNFα, and 5HT with ELISA. The hippocampal Shh-signaling pathway was assessed by Western blot. Results: EA significantly decreased sucrose preference and locomotor activities of PSD rats, reduced IL6, TNFα, increased GSH, and upregulated 5HT, and also slightly reduced IL1ß and malondialdehyde, all of which were measured with ELISA. The Shh-signaling pathway assessed by Western blotting was activated by EA. Those changes were inhibited by the Shh-pathway inhibitor cyclopamine. Conclusion: EA effectively alleviated depressive-like behaviors in PSD by suppressing inflammation and oxidative stress through activation of the Shh-signaling pathway.

4.
Ageing Res Rev ; 50: 102-109, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30711712

RESUMO

BACKGROUND: Post stroke depression is a significant neuropsychiatric manifestation, predicting a range of poor outcomes. There are several studies investigating the association between post stroke depression and stroke recurrence/mortality, but results have been inconsistent. OBJECTIVE: A systematic review, meta-analysis and meta regression of observational studies assessing the association between post stroke depression and risk of stroke recurrence and mortality. METHODS: A search of Medline (via PubMed), Web of Science databases, EMBASE, Cochrane Central Register of Controlled Trials and the Cochrane Database of Systematic Reviews was conducted until August 2018. We extracted and pooled hazard ratios from observational studies that reported the risk estimates of stroke recurrence and mortality in stroke survivors with/without depression. RESULTS: The reviewed sample comprised 15 prospective cohort studies with 250,294 participants, 139,276 cases, and follow-up periods ranging from 1 to 15 years. The meta-analysis concluded a hazard ratio for post stroke depression and all-cause mortality of 1.59 (95% CI, 1.30-1.96), but research to date has been insufficient to determine the association between post stroke depression and stroke recurrence. CONCLUSION AND RELEVANCE: Post stroke depression is associated with a significantly increased risk of mortality in stroke survivors. More researches are required on the association with stroke recurrence.


Assuntos
Depressão/etiologia , Depressão/mortalidade , Acidente Vascular Cerebral/complicações , Acidente Vascular Cerebral/mortalidade , Depressão/psicologia , Humanos , Mortalidade/tendências , Estudos Observacionais como Assunto , Estudos Prospectivos , Recidiva , Estudos Retrospectivos , Fatores de Risco , Acidente Vascular Cerebral/psicologia
5.
BMJ Open ; 8(12): e026316, 2018 12 16.
Artigo em Inglês | MEDLINE | ID: mdl-30559164

RESUMO

INTRODUCTION: A number of observational studies have indicated that poststroke depression could increase the risk of stroke outcomes. There is a meta-analysis indicating that poststroke depression is a risk factor of all-cause mortality. This paper reports the protocol for a systematic review and meta-analysis to clarify the associations of poststroke depression with stroke recurrence and mortality in order to determine whether poststroke depression is a predictor of stroke outcomes according to data extracted from relevant observational studies. METHODS AND ANALYSIS: MEDLINE, Web of Science databases, EMBASE, Cochrane Central Register of Controlled Trials, Cochrane Database of Systematic Reviews will be used to conduct the search. Published studies written in English will be included. The risk of bias for the studies included in the systematic review or meta-analysis will be assessed by the Newcastle-Ottawa Quality Assessment Scale. HRs for stroke recurrence and mortality with 95% CIs will be included as primary outcomes. Subgroup analyses and meta-regression will be performed. ETHICS AND DISSEMINATION: Ethics approval will not be needed because the data used in this systematic review will be extracted from published studies. The results of the systematic review focusing on whether depression after stroke is a predictor for stroke recurrence and mortality will be disseminated by publication in a peer-reviewed journal. PROSPERO REGISTRATION NUMBER: CRD42018107944.


Assuntos
Depressão/etiologia , Acidente Vascular Cerebral/mortalidade , Acidente Vascular Cerebral/psicologia , Adulto , Causas de Morte , Depressão/epidemiologia , Humanos , Metanálise como Assunto , Projetos de Pesquisa , Revisões Sistemáticas como Assunto
6.
J Clin Med ; 7(12)2018 Nov 24.
Artigo em Inglês | MEDLINE | ID: mdl-30477228

RESUMO

Oral squamous cell carcinoma (OSCC) is one of the most common cancer types worldwide and can be divided into three major subsites: buccal mucosal SCC (BMSCC), tongue SCC (TSCC), and lip SCC (LSCC). The autophagy marker microtubule-associated protein light chain 3B (MAP1LC3B) and adaptor sequestosome 1(SQSTM1) are widely used proteins to evaluate autophagy in tumor tissues. However, the role of MAP1LC3B and SQSTM1 in OSCC is not fully understood, particularly in certain subsites. With a tissue microarray comprised of 498 OSCC patients, including 181 BMSCC, 244 TSCC, and 73 LSCC patients, we found that the expression levels of MAP1LC3B and cytoplasmic SQSTM1 were elevated in the tumor tissues of three subsites compared with those in adjacent normal tissues. MAP1LC3B was associated with a poor prognosis only in TSCC. SQSTM1 was associated with poor differentiation in three subsites, while the association with lymph node invasion was only observed in BMSCC. Interestingly, MAP1LC3B was positively correlated with SQSTM1 in the tumor tissues of BMSCC, whereas it showed no correlation with SQSTM1 in adjacent normal tissue. The coexpression of higher MAP1LC3B and SQSTM1 demonstrated a significantly worse disease-specific survival (DSS) and disease-free survival (DFS) in patients with BMSCC and LSCC, but not TSCC. The knockdown of MAP1LC3B and SQSTM1 reduced autophagy, cell proliferation, invasion and tumorspheres of BMSCC cells. Additionally, silencing both MAP1LC3B and SQSTM1 enhanced the cytotoxic effects of paclitaxel in the tumorspheres of BMSCC cells. Taken together, MAP1LC3B and SQSTM1 might modulate autophagy to facilitate tumorigenesis and chemoresistance in OSCC, particularly in BMSCC.

7.
Dalton Trans ; 47(42): 14932-14937, 2018 Oct 30.
Artigo em Inglês | MEDLINE | ID: mdl-30252011

RESUMO

Solid-state lithium metal batteries have emerged as a promising alternative to existing liquid Li-ion batteries and can power the future storage market considering their higher energy outputs and better safety. Among various solid electrolytes, polymer electrolytes have received more attention due to their potential advantages, including wide electrochemical windows, ease of processing, low interface impedance and low cost. Polymeric electrolytes based on poly(ethylene oxide) (PEO) as a well-known polymer matrix have been extensively studied because of their highly flexible EO segments in the amorphous phase that can provide channels for lithium ion transport. However, obtaining a PEO-based solid electrolyte with high Li ion conductivity and without sacrificing mechanical strength is still a huge challenge. In this study, polymethylhydrogen-siloxane (PMHS) with low glass transition temperature and good flexibility was blended into the PEO to optimize ion transportation by the solution casting technique. The hybrid electrolyte membrane with 40% PMHS exhibited high ionic conductivity (2.0 × 10-2 S cm-1 at 80 °C), large electrochemical windows (5.2 V), a high degree of flexibility, and thermal stability. When assembling a Li/LiFePO4 battery, a reversible capacity close to 140 mA h g-1 (0.1 C) at 60 °C was delivered. In addition, a cell with this polymer electrolyte exhibits excellent stability. These results demonstrate that solid polymer electrolyte systems are eligible for next-generation high energy density all-solid-state lithium ion batteries.

8.
Chemistry ; 24(38): 9606-9611, 2018 Jul 05.
Artigo em Inglês | MEDLINE | ID: mdl-29633384

RESUMO

MnO is a promising high-capacity anode material for lithium-ion batteries (LIBs), but pristine material suffers short cycle life and poor rate capability, thus hindering the practical application. In this work, a new type of porous MnO microballs stringed with N-doped porous carbon (3DHB-MnO@NC) with a well-connected hierarchical three-dimensional network structure was prepared by the facile self-template method. The 3DHB-MnO@NC electrode can effectively promote the ion/electron transfer and buffer the large volume change of electrode during the electrochemical reaction. As the anode for LIBs, the 3DHB-MnO@NC possesses outstanding cycling performance (1247.7 mA h g-1 after 90 cycles at 200 mA g-1 ) and good rate capabilities (949.6 mA h g-1 after 450 cycles at 1000 mA g-1 ). The facile self-template method of the prepared 3DHB-MnO@NC composite paves a new way for practical applications of MnO in high performance LIBs.

9.
Nanomaterials (Basel) ; 8(2)2018 Jan 27.
Artigo em Inglês | MEDLINE | ID: mdl-29382044

RESUMO

Nanoparticle alignment on the substrate attracts considerable attention due to its wide application in different fields, such as mechanical control, small size electronics, bio/chemical sensing, molecular manipulation, and energy harvesting. However, precise nanoparticle positioning and deposition control with high fidelity are still challenging. Herein, a straightforward strategy for high quality nanoparticle-alignment by chemical lift-off lithography (CLL) is demonstrated. This technique creates high resolution self-assembled monolayer (SAM) chemical patterns on gold substrates, enabling nanoparticle-selective deposition and precise alignment. The fabricated nanoparticle arrangement geometries and dimensions are well-controllable in a large area. With proper nanoparticle surface functionality control and adequate substrate molecular manipulation, well-defined nanoparticle arrays with single-particle-wide alignment resolution are achieved.

10.
J Med Microbiol ; 67(3): 441-451, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29458539

RESUMO

PURPOSE: Avian colibacillosis is responsible for economic losses to poultry producers worldwide. To combat this, we aimed to develop an effective oral vaccine for chicken against O78 avian pathogenic Escherichia coli (APEC) infection through a Lactobacillus delivery system. METHODOLOGY: Eight Lactobacillus strains isolated from the intestines of broiler chickens were evaluated based on their in vitro adherence ability to assess their potential as a delivery vector. Fimbrial subunit A (FimA) and outer-membrane protein C (OmpC) of APEC with and without fusion to dendritic cell-targeting peptide (DCpep) and microfold cell-targeting peptide (Co1) were displayed on the surface of Lactobacillus saerimneri M-11 and yielded vaccine groups (pPG-ompC-fimA/M-11 and pPG-ompC-fimA-Co1-DCpep/M-11, respectively). The colonization of the recombinant strains in vivo was assessed and the immunogenicity and protective efficacy of orally administered recombinant strains in chickens were evaluated. RESULTS: The colonization of the recombinant strains in vivo revealed no significant differences between the recombinant and wild-type strains. Chickens orally administered with vaccine groups showed significantly higher levels of OmpC/FimA-specific IgG in serum and mucosal IgA in cecum lavage, nasal lavage and stool compared to the pPG/M-11 group. After challenge with APEC CVCC1553, better protective efficacy was observed in chickens orally immunized with pPG-ompC-fimA/M-11 and pPG-ompC-fimA-Co1-DCpep/M-11, but no significant differences were observed between the two groups. CONCLUSIONS: Recombinant chicken-borne L. saerimneri M-11 showed good immunogenicity in chickens, suggesting that it may be a promising vaccine candidate against APEC infections. However, the activity of mammalian DCpep and Co1 was not significant in chickens.


Assuntos
Infecções por Escherichia coli/veterinária , Vacinas contra Escherichia coli/imunologia , Proteínas de Fímbrias/imunologia , Imunogenicidade da Vacina , Lactobacillus/genética , Porinas/imunologia , Doenças das Aves Domésticas/imunologia , Administração Oral , Animais , Anticorpos Antibacterianos/sangue , Antígenos de Bactérias/genética , Antígenos de Bactérias/imunologia , Ceco/imunologia , Galinhas , Escherichia coli/imunologia , Escherichia coli/patogenicidade , Infecções por Escherichia coli/imunologia , Infecções por Escherichia coli/prevenção & controle , Proteínas de Fímbrias/genética , Imunoglobulina A/sangue , Imunoglobulina G/sangue , Intestinos/microbiologia , Lactobacillus/crescimento & desenvolvimento , Lactobacillus/imunologia , Lactobacillus/isolamento & purificação , Porinas/genética , Doenças das Aves Domésticas/prevenção & controle
11.
Can J Microbiol ; 64(2): 147-154, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-29220592

RESUMO

The increased serum survival gene (iss) has been identified as a virulence trait associated with the virulence of Escherichia coli, causing colibacillosis in poultry. However, it remains unclear as to whether iss mRNA copy number and sequence affect virulence. To examine these influences, we assessed the presence of iss, sequence analysis, iss mRNA copy number, and serum resistance. The iss gene was detected in 88 (all) E. coli isolates from different sources, and sequencing identified 16 alleles (32 different loci) and 10 amino acid sequences (10 different loci). Nested polymerase chain reaction improved iss detection. The isolates from sick chickens had >68% livability in serum resistance tests and higher iss mRNA copy number. The iss mRNA copy number highly correlated with mortality and E. coli livability. Student's t tests confirmed the relationship between the different loci to iss transcription, serum resistance, and virulence. These data suggest that iss mRNA copy number and different loci affect the virulence and serum resistance. These findings could be useful in further studies on the prevalence of iss among E. coli isolates and other virulence factors.


Assuntos
Infecções por Escherichia coli/veterinária , Proteínas de Escherichia coli/genética , Escherichia coli/genética , Escherichia coli/patogenicidade , Doenças das Aves Domésticas/microbiologia , RNA Mensageiro/genética , Virulência/genética , Sequência de Aminoácidos , Animais , Galinhas , Infecções por Escherichia coli/microbiologia , Infecções por Escherichia coli/mortalidade , Dosagem de Genes , Reação em Cadeia da Polimerase , Aves Domésticas , Doenças das Aves Domésticas/mortalidade , Fatores de Virulência/genética
12.
J Virol Methods ; 243: 80-82, 2017 05.
Artigo em Inglês | MEDLINE | ID: mdl-28185831

RESUMO

In this study, a specific and sensitive method for simultaneous detection of human astrovirus, human rotavirus, norovirus, sapovirus and enteric adenovirus associated with acute enteritis was developed, based on the specific dual priming oligonucleotide (DPO) system and the sensitive high-performance liquid chromatography (HPLC) analysis. The DPO system-based multiplex reverse transcription-polymerase chain reaction (RT-PCR) combined with HPLC assay was more sensitive than agarose gel electrophoresis analysis and real-time SYBR Green PCR assay, and showed a specificity of 100% and sensitivity of 96%-100%. The high sensitivity and specificity of the assay indicates its great potential to be a useful tool for the accurate diagnosis of enteric virus infections.


Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Enterite/diagnóstico , Técnicas de Diagnóstico Molecular/métodos , Reação em Cadeia da Polimerase Multiplex/métodos , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Viroses/diagnóstico , Humanos , Sensibilidade e Especificidade
13.
Addict Biol ; 22(2): 435-445, 2017 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26692025

RESUMO

The glutamatergic projection from the ventral subiculum of the hippocampus (vSUB) to the nucleus accumbens (NAc) shell has been reported to play a key role in drug-related behavior. The GluN2B subunit of N-methyl-D-aspartate receptors (NMDARs) in the NAc can be selectively elevated after the retrieval of drug-conditioned memory. However, whether the increased GluN2B-containing NMDARs (GluN2B-NMDARs) are able to alter the synaptic plasticity of the vSUB-NAc glutamatergic pathway remains unclear. Here, we found that the long-term potentiation (LTP) in the vSUB-NAc pathway was facilitated and the GluN2B subunit protein level was elevated in synaptoneurosomes of the NAc shell, but not in the core, following morphine-induced conditioned place preference (CPP) expression in rats. The facilitated LTP was prevented by the GluN2B-NMDAR antagonist RO25-6981. Also, a neurochemical disconnection following microinjection of RO25-6981 into the NAc shell, plus microinfusion of GABA agonist baclofen and muscimol into the contralateral vSUB prevented the expression of morphine-induced CPP. These findings suggest that the retrieval of drug-associated memory potentiated synaptic plasticity in the vSUB-NAc pathway, which was dependent on GluN2B-NMDAR activation in the NAc shell. These findings provide a new explanation for the mechanisms that underlie the morphine-associated-context memory. The GluN2B-NMDARs may be regarded as a potential target for erasing morphine-related memory.


Assuntos
Analgésicos Opioides/farmacologia , Comportamento Animal/efeitos dos fármacos , Condicionamento Operante , Hipocampo/efeitos dos fármacos , Potenciação de Longa Duração/efeitos dos fármacos , Morfina/farmacologia , Núcleo Accumbens/efeitos dos fármacos , Receptores de N-Metil-D-Aspartato/efeitos dos fármacos , Animais , Baclofeno/farmacologia , Agonistas de Receptores de GABA-A/farmacologia , Agonistas dos Receptores de GABA-B/farmacologia , Hipocampo/metabolismo , Masculino , Memória/efeitos dos fármacos , Muscimol/farmacologia , Núcleo Accumbens/metabolismo , Fenóis/farmacologia , Piperidinas/farmacologia , Ratos , Ratos Sprague-Dawley , Receptores de N-Metil-D-Aspartato/antagonistas & inibidores , Receptores de N-Metil-D-Aspartato/metabolismo
14.
Appl Microbiol Biotechnol ; 100(22): 9593-9604, 2016 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-27557715

RESUMO

Using two-step plasmid integration in the presence of 5-fluorouracil (5-FU), we developed a stable and markerless Lactobacillus casei strain for vaccine antigen expression. The upp of L. casei, which encodes uracil phosphoribosyltransferase (UPRTase), was used as a counterselection marker. We employed the Δupp isogenic mutant, which is resistant to 5-FU, as host and a temperature-sensitive suicide plasmid bearing upp expression cassette as counterselectable integration vector. Extrachromosomal expression of UPRTase complemented the mutated chromosomal upp allele and restored sensitivity to 5-FU. The resultant genotype can either be wild type or recombinant. The efficacy of the system was demonstrated by insertion and expression of porcine rotavirus (PRV) VP4. To improve VP4 expression, we analyzed L. casei transcriptional profiles and selected the constitutive highly expressed enolase gene (eno). The VP4 inserted after the eno termination codon were screened in the presence of 5-FU. Using genomic PCR amplification, we confirmed that VP4 was successfully integrated and stably inherited for at least 50 generations. Western blot demonstrated that VP4 was steadily expressed in medium with different carbohydrates. RT-qPCR and ELISA analysis showed that VP4 expression from the chromosomal location was similar to that achieved by a plasmid expression system. Applying the recombinant strain to immunize BALB/c mice via oral administration revealed that the VP4-expressing L. casei could induce both specific local and systemic humoral immune responses in mice. Overall, the improved gene replacement system represents an efficient method for chromosome recombination in L. casei and provides a safe tool for vaccine production.


Assuntos
Proteínas do Capsídeo/biossíntese , Expressão Gênica , Marcação de Genes/métodos , Lactobacillus casei/genética , Proteínas Recombinantes/biossíntese , Vacinas contra Rotavirus/imunologia , Administração Oral , Animais , Proteínas do Capsídeo/genética , Instabilidade Genômica , Camundongos Endogâmicos BALB C , Proteínas Recombinantes/genética , Recombinação Genética , Vacinas contra Rotavirus/administração & dosagem , Vacinas contra Rotavirus/genética , Vacinas Sintéticas/administração & dosagem , Vacinas Sintéticas/genética , Vacinas Sintéticas/imunologia
15.
Neuropharmacology ; 102: 216-28, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26596557

RESUMO

The nitric oxide (NO)/soluble guanylyl cyclase (sGC)/cGMP-dependent protein kinase (PKG) signaling pathway has been reported to play a key role in memory processing. However, little is known about its role in drug-associated reward memory. Here, we report the following. 1) The NO pathway in the CA1 is critical for the retrieval of morphine-associated reward memory. Specifically, the nNOS, sGC and PKG protein levels in the CA1 were increased after the expression of morphine conditioned place preference (CPP). Intra-CA1 injection of an NOS, sGC or PKG inhibitor prevented morphine CPP expression. 2) The involvement of the NO pathway in morphine CPP requires NR2B-containing NMDA receptors (NR2B-NMDARs). NR2B-NMDAR expression was elevated in the CA1 following morphine CPP expression, and intra-CA1 injection of the NR2B-NMDAR antagonist Ro25-6981 not only blocked morphine CPP expression but also inhibited the up-regulation of nNOS, sGC and PKG. Moreover, the Ro25-6981-induced blockade of morphine CPP was abolished by intra-CA1 injection of a NOS substrate or an sGC activator. 3) The NR2B-NMDAR stimulated the NO pathway by up-regulating the phosphorylation of Akt(Ser473). Morphine CPP expression enhanced the pAkt(Ser473) level, which has been corroborated to regulate nNOS activity, and this effect was reversed by intra-CA1 injection of Ro25-6981. 4) GluR1 acted downstream of the NO pathway. The membrane level of GluR1 in the CA1 was increased after morphine CPP expression, and this effect was prevented by pre-injection of a PKG inhibitor into the CA1. Additionally, co-immunoprecipitation revealed an interaction between PKG and GluR1; this result further indicated a role of PKG in regulating GluR1 trafficking. Collectively, the results of our study demonstrated that the activation of the NR2B-NMDAR/NO/sGC/PKG signaling pathway is necessary for the retrieval of morphine-associated reward memory.


Assuntos
Região CA1 Hipocampal/metabolismo , Condicionamento Operante/efeitos dos fármacos , Memória/efeitos dos fármacos , Morfina/farmacologia , Óxido Nítrico/metabolismo , Receptores de Neurotransmissores/metabolismo , Transdução de Sinais/fisiologia , Animais , Aprendizagem por Associação/efeitos dos fármacos , Aprendizagem por Associação/fisiologia , Região CA1 Hipocampal/efeitos dos fármacos , Condicionamento Operante/fisiologia , Masculino , Memória/fisiologia , Fosforilação/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-akt/metabolismo , Ratos , Ratos Sprague-Dawley , Recompensa , Transdução de Sinais/efeitos dos fármacos , Regulação para Cima/efeitos dos fármacos
16.
Proteomics ; 15(11): 1843-58, 2015 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-25684102

RESUMO

Similar to the well-studied viruses human immunodeficiency virus (HIV)-1 and simian immunodeficiency virus (SIV), equine infectious anemia virus (EIAV) is another member of the Lentivirus genus in the family Retroviridae. Previous studies revealed that interactions between EIAV and the host resulted in viral evolution in pathogenicity and immunogenicity, as well as adaptation to the host. Proteomic analysis has been performed to examine changes in protein expression and/or modification in host cells infected with viruses and has revealed useful information for virus-host interactions. In this study, altered protein expression in equine monocyte-derived macrophages (eMDMs, the principle target cell of EIAV in vivo) infected with the EIAV pathogenic strain EIAV(DLV34) (DLV34) was examined using 2D-LC-MS/MS coupled with the iTRAQ labeling technique. The expression levels of 210 cellular proteins were identified to be significantly upregulated or downregulated by infection with DLV34. Alterations in protein expression were confirmed by examining the mRNA levels of eight selected proteins using quantitative real-time reverse-transcription PCR, and by verifying the levels of ten selected proteins using parallel reaction monitoring (PRM). Further analysis of GO and Kyoto Encyclopedia of Genes and Genomes (KEGG)-Pathway enrichment demonstrated that these differentially expressed proteins are primarily related to the biological processes of oxidative phosphorylation, protein folding, RNA splicing, and ubiquitylation. Our results can facilitate a better understanding of the host response to EIAV infection and the cellular processes required for EIAV replication and pathogenesis.


Assuntos
Interações Hospedeiro-Patógeno , Vírus da Anemia Infecciosa Equina/patogenicidade , Macrófagos/metabolismo , Macrófagos/virologia , Proteoma/análise , Sequência de Aminoácidos , Animais , Células Cultivadas , Anemia Infecciosa Equina/metabolismo , Ontologia Genética , Cavalos , Dados de Sequência Molecular , Proteoma/genética , Proteoma/metabolismo , Reprodutibilidade dos Testes , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Espectrometria de Massas em Tandem , Replicação Viral
17.
Behav Neurosci ; 128(4): 446-59, 2014 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-25046820

RESUMO

There is evidence that the nitric oxide (NO)/soluble guanylyl cyclase (sGC)/cGMP-dependent protein kinase (PKG) signaling pathway in the basal lateral amygdala and hippocampus plays a key role in memory processing, but it is not known if this NO signaling pathway in the nucleus accumbens (Gomes et al., 2006), a known pivotal region in reward memory, is essential for drug-associated reward memory. We therefore investigated the effect of the NO/sGC/PKG signaling pathway in the nucleus accumbens (NAc) on morphine-induced conditioned place preference (CPP). Results showed that a preconditioning microinjection of the NO synthase (NOS) inhibitor Nω-nitro-L-arginine methyl ester (L-NAME) into the NAc shell, but not into the core, significantly blocked the acquisition of morphine CPP. The blockage effect of L-NAME on the acquisition of CPP was imitated by the neuronal NOS inhibitor 7-nitroindazole, 3-bromo-, sodium salt (7-NI), the sGC inhibitor 1H-[1,2,4] oxadiazolo-[4,3-a]quinoxalin-1-one (ODQ), and the PKG inhibitor Rp-8Br-PET-cGMPS. The 7-NI- or ODQ-induced effect was reversed by premicroinjection of the sGC activator YC-1 or the PKG activator 8-Br-cGMP in the NAc shell. However, microinfusion of 7-NI, ODQ, or Rp-8Br-PET-cGMPS into the NAc shell or the core had no effect on the expression of morphine CPP. These findings indicate that the NO/sGC/PKG signaling pathway in the NAc shell is critical for the acquisition of morphine-induced place preference, whereas the same signaling pathway in the NAc shell or core is not involved in the retrieval of morphine-induced place preference.


Assuntos
Condicionamento Clássico/efeitos dos fármacos , Proteínas Quinases Dependentes de GMP Cíclico/metabolismo , Guanilato Ciclase/metabolismo , Morfina/farmacologia , Óxido Nítrico/metabolismo , Núcleo Accumbens/metabolismo , Receptores Citoplasmáticos e Nucleares/metabolismo , Transdução de Sinais , Animais , Condicionamento Clássico/fisiologia , Masculino , Ratos , Ratos Sprague-Dawley , Guanilil Ciclase Solúvel
18.
Exp Appl Acarol ; 63(4): 579-85, 2014 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-24699771

RESUMO

Since the year 2000, clinical patterns resembling tick-borne rickettsioses have been noticed in China-Russia border areas. Epidemiological data regarding species of the aetiological agent, tick vector prevalence and distribution as well as incidence of human cases in the areas are still sparse to date. In order to identify Rickettsia species occurring in the areas, we investigated Dermacentor silvarum collected in the selected areas. Rickettsia raoultii was the predominant Rickettsia found in D. silvarum evident with ompA, ompB, gltA and 17 kDa protein genes. The Rickettsia prevalence in D. silvarum appeared to be 32.25 % with no sex difference. The results extend the common knowledge about the geographic distribution of R. raoultii and its candidate vector tick species, which suggest an emerged potential threat of human health in the areas.


Assuntos
Dermacentor/microbiologia , Filogenia , Infecções por Rickettsia/microbiologia , Rickettsia/isolamento & purificação , Doenças Transmitidas por Carrapatos/microbiologia , Animais , Proteínas da Membrana Bacteriana Externa/química , Proteínas da Membrana Bacteriana Externa/genética , Sequência de Bases , China/epidemiologia , Citrato (si)-Sintase/química , Citrato (si)-Sintase/genética , DNA Bacteriano/química , DNA Bacteriano/genética , Feminino , Humanos , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Rickettsia/genética , Infecções por Rickettsia/epidemiologia , Federação Russa/epidemiologia , Alinhamento de Sequência , Análise de Sequência de DNA , Doenças Transmitidas por Carrapatos/epidemiologia
19.
Appl Environ Microbiol ; 80(11): 3321-6, 2014 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-24657853

RESUMO

To develop a stable and marker-free Lactobacillus strain useful for the expression of vaccines, we developed a temperature-sensitive suicide plasmid with expression cassettes containing an HCE promoter, a PgsA anchor, the alpha-toxin gene, and an rrnB T1T2 terminator (PPαT) that uses a 5-fluorouracil (5-FU) counterselectable marker for Lactobacillus casei. Three strains containing the correct PPαT expression cassettes were produced via the selective pressure of 5-FU screening. We confirmed that the upp gene was deleted and that the PPαT expression cassettes were inserted into the upp site of L. casei ATCC 393 by genomic PCR amplification and sequencing. 5-FU resistance in recombinant bacteria could be stably inherited for as long as 40 generations following insertion. However, bacteria containing the integrated DNA grew more slowly than wild-type L. casei. An indirect enzyme-linked immunosorbent assay (ELISA) analysis demonstrated that the alpha-toxin gene was expressed. Also, we visualized expression of the protein on the surface of L. casei cells using laser confocal microscopy. These results taken together demonstrate that these recombinant bacteria should provide a safe tool for effective vaccine production.


Assuntos
Antígenos/genética , Antígenos/imunologia , Vacinas Bacterianas/genética , Vacinas Bacterianas/imunologia , Lactobacillus casei/genética , Mutagênese Insercional , Técnicas de Visualização da Superfície Celular , Proteínas de Membrana/genética , Proteínas de Membrana/imunologia , Biologia Molecular/métodos , Plasmídeos
20.
Int Rev Neurobiol ; 111: 235-56, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-24215926

RESUMO

Over the last four decades, there has been an increasing interest in acupuncture treatment of substance abuse around the world. Three important steps can be identified in this field. Dr. Wen of Hong Kong was the first (1972) to report that acupuncture at four body points and two ear points combined with electric stimulation can relieve opioid withdrawal signs in the addicts. The second major step was made by Dr. M. Smith in New York, the head of the National Acupuncture Detoxification Association (NADA) of the United States, who finalized a protocol (1985), using only ear points without electric stimulation for the treatment of cocaine dependence. The recent advance in this field was made by Dr. Han of the Peking University, Beijing, who characterized a protocol (2005), using electric stimulation of identified frequencies on body points to ameliorate heroin withdrawal signs and reduce relapse of heroin use. In this chapter, the efficacy of acupuncture and related techniques for the treatment of drug dependence in experimental settings and clinical practice will be reviewed, and the possible mechanisms underlying this effect be discussed.


Assuntos
Terapia por Acupuntura , Transtornos Relacionados ao Uso de Substâncias/terapia , Humanos
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