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Current knowledge of RNA virus biodiversity is both biased and fragmentary, reflecting a focus on culturable or disease-causing agents. Here we profile the transcriptomes of over 220 invertebrate species sampled across nine animal phyla and report the discovery of 1,445 RNA viruses, including some that are sufficiently divergent to comprise new families. The identified viruses fill major gaps in the RNA virus phylogeny and reveal an evolutionary history that is characterized by both host switching and co-divergence. The invertebrate virome also reveals remarkable genomic flexibility that includes frequent recombination, lateral gene transfer among viruses and hosts, gene gain and loss, and complex genomic rearrangements. Together, these data present a view of the RNA virosphere that is more phylogenetically and genomically diverse than that depicted in current classification schemes and provide a more solid foundation for studies in virus ecology and evolution.
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Ubiquitylation is an essential type of protein post-translational modifications (PTMs) in eukaryotes, which mediates various biological processes by regulating the subcellular localization, activity, and stability of proteins. Histones, as the main protein ingredients of chromatin, are closely coupled with DNA activities such as replication, transcription and repair, and therefore are the hotspots of PTMs. After DNA damage, histone ubiquitylations are involved in DNA damage response (DDR) by regulating nucleosome structure, activating cell cycle checkpoints, remodeling the nucleosome, and the recruitment and assembly of repair factors. Meanwhile, histone ubiquitylations can also crosstalk with other types of PTMs to regulate DDR processes. In this review, we summarize how the site-specific histone ubiquitylation forms signal network and contributes to DDR, which may shed light on the further study of how histone codes formed by histone PTMs affect the entire DDR processes.
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Daño del ADN , Histonas/química , Ubiquitinación , Cromatina , Reparación del ADN , HumanosRESUMEN
BACKGROUND: Excretory-secretory products released by Echinococcus granulosus protoscoleces (EgPSC-ESPs) are well-known to regulate T cell responses. However, their direct influence on the differentiation of B cell subsets remains largely elusive. This study investigated the effects of EgPSC-ESPs on the differentiation of IL-10-producing B cells (B10), and explored the possible role of Toll-like receptor 2 (TLR-2) signaling in this process. RESULTS: In comparison to phosphate buffered saline (PBS), B cells exposed to the excretory-secretory products (ESPs) generated higher percentages of B10 cells, with higher expression of IL-10 mRNA, and larger amount of IL-10 production, which were in a dose dependent way. The mRNA and protein expression of TLR-2 in the ESPs-stimulated B cells were significantly higher than those in PBS, which was consistent to the results in B cells isolated from EgPSC infected mice. Moreover, TLR-2-/- B cells in response to ESPs stimulation expressed lower levels of IL-10 mRNA and produced undetectable IL-10 in comparison to those in normal B cells. In addition, Phosphatase and tensin homolog deleted on chromosome ten/AKT/Phosphatidylinositol-3 kinase (PTEN/AKT/PI3K) pathway was activated in ESPs-treated B cells, which was also dependent on TLR-2 signaling. Pam3CSK4, the agonist of TLR-2, could mock the effects of ESPs on the expression of PTEN, AKT and PI3K. CONCLUSION: Overall, this study revealed that TLR-2 signaling was required for B10 induction mediated by EgPSC-ESPs, which might be an immunomodulatory target against the parasite infection.
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Antígenos Helmínticos/inmunología , Subgrupos de Linfocitos B/inmunología , Equinococosis/inmunología , Echinococcus granulosus/inmunología , Interleucina-10/metabolismo , Receptor Toll-Like 2/metabolismo , Animales , Interleucina-10/genética , Ratones Endogámicos C57BL , Fosfohidrolasa PTEN/metabolismo , Fosfatidilinositol 3-Quinasa/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Receptor Toll-Like 2/genéticaRESUMEN
BACKGROUND/AIMS: This study aims to predict the pro-angiogenic functions of monocytic-type myeloid-derived suppressor cells (M-MDSCs) derived from mice infected with Echinococcus granulosus. METHODS: M-MDSCs were collected from Balb/c mice infected with E. granulosus and normal mice (control) and cultured in vitro. Human umbilical vein endothelial cells (HUVECs) were stimulated with the cell supernatant, and angiogenesis was investigated and analysed by the Angiogenesis module of the software NIH Image J. RNA was extracted from fresh isolated M-MDSCs and analysed with miRNA microarray; differentially expressed miRNAs and their potential functions were analysed through several bioinformatics tools. Finally, quantitative PCR was used to confirm the results of microarray analysis. RESULTS: M-MDSCs from mice infected with E. granulosus could promote the formation of tubes from HUVECs in vitro. Moreover, vascular endothelial growth factor (VEGF) showed significantly high expression, whereas soluble fms-like tyrosine kinase-1 (sFlt-1) showed low expression at the transcriptional level in M-MDSCs from mice infected with E. granulosus. Microarray analysis of miRNAs showed that 28 miRNAs were differentially expressed in M-MDSCs from the two experimental mice groups, and 272 target genes were predicted using the microRNA databases TargetScan, PITA and microRNAorg. These target genes were mainly involved in the biological processes of intracellular protein transport, protein targeting to the lysosome and protein transport, and mainly located in the cytoplasm, neuronal cell body and membrane. Moreover, they were mainly involved in the molecular functions of protein binding, metal ion binding and SH3 domain binding. Further, the differentially expressed miRNAs were mainly enriched in the endocytosis, Wnt and axon guidance pathways, as well as the MAPK, focal adhesion, PI3K-Akt, cAMP, mTOR and TGF-ß signalling pathways, which are linked to immunoregulation and angiogenesis based on the results of bioinformatics analysis with DIANA-miRPath 3.0. In addition, the expression of eight miRNAs was randomly verified by quantitative PCR independently in three mice infected with E. granulosus and three normal mice. CONCLUSION: M-MDSCs have a potential angiogenic role during E. granulosus infection, and miRNAs may play a role in the immune response and angiogenesis functions of M-MDSCs through regulation of the identified signalling pathways.
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Equinococosis/genética , Echinococcus granulosus/fisiología , Regulación de la Expresión Génica , MicroARNs/genética , Células Supresoras de Origen Mieloide/virología , Neovascularización Patológica/genética , Animales , Células Cultivadas , Equinococosis/patología , Equinococosis/virología , Células Endoteliales de la Vena Umbilical Humana , Humanos , Ratones , Ratones Endogámicos BALB C , Células Supresoras de Origen Mieloide/patología , Neovascularización Patológica/patología , Neovascularización Patológica/virología , Factor A de Crecimiento Endotelial Vascular/genéticaRESUMEN
Objective: To investigate the alteration of expression and activity of arginase from monocytic-type myeloid-derived suppressor cellsï¼M-MDSCï¼ in BALB/c mice infected with Echinococcus granulosus. Methods: Twelve BALB/c female mice were randomly divided into control and infected groups. The mice were injected intraperitoneally with 2 000 live protoscoleces or an equivalent volume of normal saline. After 120 days, peripheral blood was collected through venae orbitaeta, and mice were sacrificed for pathological examination. The spleen was collected under aseptic conditions and single-cell suspension was prepared for M-MDSC isolation using the magnetic bead separation technology. Total RNA was extracted from M-MDSC, cDNA was generated, and genes with differential expression without and with infection were screened using the chip hybridization method. The resulting genes were further validated using real-time PCR. The activity of arginase from peripheral blood was also measured. Results: Single cyst was formed within the abdomen and internal organs 120 days after infection. Chip hybridization and real-time PCR showed that the relative expression of arginase from M-MDSC in the infected group ï¼7.92±0.85 and 11.97±5.39, respectivelyï¼ was significantly higher than that in the control group ï¼1.65±0.19 and 1.00±0.57, respectivelyï¼ ï¼P<0.05ï¼. The activity of arginase was also significantly higher in the infected group ï¼»ï¼3.83±0.44ï¼U/Lï¼½ than in the control [(1.57±0.57ï¼U/L]. Conclusion: The expression and activity of arginase from mouse M-MDSC both increase significantly after infection with Echinococcus granulosus.
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Echinococcus granulosus , Animales , Arginasa , Femenino , Ratones , Ratones Endogámicos BALB C , Monocitos , Células Supresoras de Origen Mieloide , Ratas , BazoRESUMEN
Objective: To examine the IgG and IgM antibodies for parasites Cysticercus cellulosae and Toxoplasma gondii in 122 patients with meningitis encephalitis syndrome, and provide basis for clinical diagnosis of the meningitis encephalitis syndrome. Methods: The sera were collected from patients with meningitis encephalitis syndrome in Shanghai Jiaotong University Affiliated Sixth People's Hospital, People's Hospital of Danyang City, and Jiangsu University Affiliated Hospital from August, 2014 to December, 2015. Serum IgG and IgM antibodies for cysticercus and T. gondii were examined using antibody test kits. The antibody positive rate was calculated and its distribution was analyzed by gender, season, age and occupation. Results: A total of 122 patients with meningitis encephalitis syndrome were included. Seventeen and 22 patients of them were positive for IgG ï¼13.9%, 17/122ï¼ and IgMï¼18.0%, 22/122ï¼ against cysticercus, respectively, while 29 and 8 cases were positive for IgG ï¼23.8%, 29/122ï¼ and IgM ï¼6.6%, 8/122ï¼ against T. gondii. The positive rate of cysticercus and T. gondii in males was 30.6%ï¼22/72ï¼ and 31.9%ï¼23/72ï¼ respectively, while that in females was 26.0%ï¼13/50ï¼ and 24.0% ï¼12/50ï¼. The positive rate of IgM against cysticercus was 12.0%ï¼3/25ï¼, 27.0%ï¼17/63ï¼, 6.9% ï¼2/29ï¼, and 0ï¼0/5ï¼ from spring to winter, highest within 13-25 yearsï¼45.0%, 9/20ï¼ among age groups, and highest in workersï¼7/14ï¼ among various occupations. The positive rate of IgM against T. gondii was 4.0%ï¼1/25ï¼, 11.1% ï¼7/63ï¼, 0ï¼0/29ï¼, and 0ï¼0/5ï¼ from spring to winter, highest in ages >65 yearsï¼44.0%, 11/25ï¼, and highest in patients with other occupationsï¼4/10ï¼. There was no statistically significant difference in the positive rate between males and females, and among different seasons, ages and occupations. Conclusion: The positive rate of antibodies against cysticercus and T. gondii is high in the patients included, suggesting that a serological test for parasite infection might be performed during clinical diagnosis of meningitis encephalitis syndrome.
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Meningitis , Toxoplasma , Toxoplasmosis , Adolescente , Adulto , Anciano , Animales , Anticuerpos Antiprotozoarios , China , Cysticercus , Encefalitis , Femenino , Humanos , Inmunoglobulina G , Inmunoglobulina M , Pruebas Inmunológicas , Masculino , Enfermedades Parasitarias , Taenia solium , Adulto JovenRESUMEN
Objective: To investigate the phenotype and phagocytosis changes of the peritoneal macrophages (Mφ) in mice infected with the larval-stage Echinococcus granulosus, and explore the role of Mφ in the responses to parasite infection. Methods: Twenty-four female BALB/c mice (age of 6-8 weeks) were randomly assigned into control group and infection group (n=12 in each group). The mice in the infection group were intraperitoneally injected with 2 000 protoscoleces, while the control mice were injected with equal volume of PBS. Five months after infection, the peritoneal mononuclear cells were collected, and the percentage of Mφ and the expression of surface markers CD40, CD80, CD86, and major histocompatibility complex â ¡ (MHCâ ¡) were determined by flow cytometry. The absorbanceï¼A490 valueï¼ of Mφ at different concentrationsï¼1×106, 5×105, 1×105ï¼ was determined by the neutral red assay to evaluate the phagocytic ability of Mφ. Results: The Mφ constitutedï¼30.40±3.15ï¼% andï¼20.75±5.91ï¼% in mononuclear cells in the infection and the control groups, respectively. The percentages of Mφ expressing CD40, CD80, CD86, and MHC â ¡ wereï¼45.33±5.51ï¼%, ï¼61.00±10.61ï¼%, ï¼56.88±10.66ï¼% and ï¼27.00±3.82ï¼% in the infection group, which were all significantly higher than those in the control ï¼»ï¼41.43±6.19ï¼%, ï¼59.23±8.65ï¼%, ï¼10.91±1.82ï¼% and ï¼13.67±3.01%ï¼ï¼½ ï¼P<0.05ï¼. The A490 values of Mφ at 1×106, 5×105, 1×105 were 0.41±0.03ï¼ 0.24±0.05 and 0.16±0.01 in the infection group, which were significantly lower than those in the control ï¼0.61±0.15, 0.47±0.07 and 0.18±0.01ï¼ï¼P<0.01ï¼. Conclusion: The phagocytic ability of peritoneal Mφ is dramatically weakened after infection, but the expression of activation-associated surface markers is significantly up-regulated after infection.
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Echinococcus granulosus , Macrófagos Peritoneales , Animales , Femenino , Citometría de Flujo , Larva , Macrófagos , Ratones , Ratones Endogámicos BALB C , Fagocitosis , FenotipoRESUMEN
Objective: To investigate the pathological changes of liver and spleen of mice infected with Schistosoma japonicum and the changes of T follicular helper (Tfh) cells and surface molecules after praziquantel treatment. Methods: Fifteen female C57BL/6 mice ï¼6-8 weeksï¼ were randomly assigned into the praziquantel treated infection group ï¼treated groupï¼, infection control group (untreated group) and uninfected group ï¼n=5 in each groupï¼. The mice in the treated group and untreated group were each infected with 20 S. japonicum cercariae through the abdominal skin, and mice in the treated group were further administered with intragastric praziquantel [200 mg/ï¼kg·dï¼] at week 6 post-infection for 3 consecutive days. Mice were sacrificed at week 4 after treatment to observe the morphological changes of liver and spleen and calculate the worm reduction rate and the liver egg reduction rate. The Tfh cell to CD4+ T cell ratio, as well as the expression of inducible T-cell costimulator ï¼ICOSï¼ and programmed cell death protein 1ï¼PD-1ï¼ on peripheral blood and spleen, were determined by flow cytometry. Schistosome soluble egg antigen (SEA) specific IgG antibodies in serum were detected by ELISA. Results: The pathological changes of liver and spleen in the treated group were less severe compared with those of the untreated group, with a worm reduction rate of 84.1% and liver egg reduction rate of 69.1%. Flow cytometry showed that the percent of Tfh cells in peripheral blood and spleen was significantly higher in the treated groupï¼14.7%-18.0%, 15.6%-25.0%ï¼ and the untreated groupï¼13.7%-16.7%, 12.4%-18.2%ï¼ than that in the uninfected groupï¼2.5%-6.8%, 4.9%-8.0%ï¼, but there was no significant difference between the treated and untreated groups. The expression of ICOS in the peripheral blood and the spleen was significantly higher in untreated groupï¼1.3%-3.2%, 4.1%-7.0%ï¼ than in the treated groupï¼0.7%-1.1%, 1.8%-6.8%ï¼ and the uninfected groupï¼0.2%-0.3%, 0.5%-0.8%ï¼ï¼P<0.01ï¼, The expression of ICOS in the spleen was significantly higher in the treated group than in the uninfected group (P<0.01), while this difference was not found for ICOS expression in the peripheral blood. The PD-1 expression in the peripheral blood and spleen was significantly higher in the untreated groupï¼0.8%-1.9%, 4.1%-10.7%ï¼ than in the uninfected groupï¼0.4%-0.8%, 1.2%-1.8%ï¼ï¼P<0.01ï¼, while there was no significant difference between the treated groupï¼0.5%-1.5%, 4.5%-8.9%ï¼ and the untreated group (P>0.05). In addition, there was no significant difference in the level of SEA specific IgG between the treated groupï¼2.015±0.061ï¼ and the untreated groupï¼1.969±0.038ï¼ at 4 weeks after praziquantel treatment. Conclusion: Praziquantel treatment can significantly alleviate the lesions of the liver and the spleen and decrease the ICOS expression by Tfh cells in the peripheral blood and spleen.
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Schistosoma japonicum , Animales , Anticuerpos Antihelmínticos , Cercarias , Ensayo de Inmunoadsorción Enzimática , Femenino , Ratones , Ratones Endogámicos C57BL , Praziquantel , Esquistosomiasis Japónica , Bazo , Linfocitos T Colaboradores-InductoresRESUMEN
Autophagy is an evolutionarily well conserved cellular catabolic process.Cellular proteins and organelles are engulfed to autophagosomes and eventually delivered to lysosomes for degradation. Autophagy is closely associated with a variety of human diseases especially cancer. At present, it has been widely accepted that autophagy is a "double-edged sword" in cancer: it blocks tumorigenesis at the early stage, while it facilitates tumor development at the promotion and progression stage. Moreover, autophagy can be induced by chemotherapy and radiotherapy, which is generally play a pro-survival function. Thus, autophagy is an important target for cancer therapy, and suppression of autophagy to enhance the efficacy of cancer therapeutic agents is a novel strategy in cancer therapy under active clinical trials.
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Autofagia , Neoplasias , Transformación Celular Neoplásica , HumanosRESUMEN
OBJECTIVE: To clone and express the profilin (PRF) gene of Toxoplasma gondii, and analyze the immunoreactivity. METHODS: Total RNA was extracted from tachyzoites of T. gondii RH strain. The coding region of TgPRF was amplified with a pair of specific primers. PCR product was digested with double restriction enzyme and ligated into pET30a(+) vector. The recombinant pET30a(+)-TgPRF plasmid was transformed into E. coli DH5α with positive clones confirmed by the double restriction enzyme digestion, PCR and sequencing. The correct plasmid was transformed into E. coli BL21 and induced by IPTG. The expressed proteins were purified with Ni-NTA affinity chromatography and analyzed by SDS-PAGE. Western blotting with rabbit anti-T. gondii serum was used to analyze its antigenicity. RESULTS: The product of RT-PCR was with 492 bp. pET30a-TgPRF was confirmed by the double restriction enzyme digestion, PCR and sequencing. SDS-PAGE analysis showed that the expressed product was a soluble protein with a relative molecular weight of 35,000. Western blotting assay revealed that rTgPRF was recognized by rabbit anti-T. gondii serum. CONCLUSION: TgPRF gene has been expressed in prokaryotic expression system and shows immunoreactivity.
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Profilinas/inmunología , Proteínas Protozoarias/inmunología , Toxoplasma , Western Blotting , Cromatografía de Afinidad , Clonación Molecular , Electroforesis en Gel de Poliacrilamida , Escherichia coli , Expresión Génica , Vectores Genéticos , Plásmidos , Reacción en Cadena de la Polimerasa , Proteínas Recombinantes/inmunologíaRESUMEN
OBJECTIVE: To construct a cocktail DNA vaccine that expresses multiple genes of Toxoplasma gondii and investigate its immunogenicity in mice. METHODS: Genes for surface antigens (SAG), microneme(MIC), and rhoptry protein(ROP) were amplified from genomic DNA of T. gondii and then cloned separately into eukaryotic fluorescent protein expression vectors pShuttle-CMV-MCS-EFlα-AmCyan, pLVX-IRES-Zsgreen and pLVX-IRES-rfp, to construct expression plasmids pShuttle-SAG1, pLVX-Zsgreen-MIC3 and pLVX-rfp-ROP2. 293F cells were transfected with a combination of the three plasmids using the polyethylenimine (PEI) method. Forty-eight hours later, the expression of the three genes was observed under a fluorescence microscope. In addition, 30 C57BL/6 mice were randomized to receive intramuscular injection of saline (50 pA, group A), pShuttle+pLVX-Zsgreen+pLVX-rfp empty plasmids(2 µg/µL, 17 µL of each, group B) and pShuttle-SAG1+pLVX-Zsgreen-MIC3+ pLVX-rfp-ROP2 recombinant plasmid (2 µg/µL, 17 µL of each, group C). After 28 days, anti-T. gondii antibody in mouse serum was detected by ELISA, to evaluate the immunogenicity of the vaccine. RESULTS: The SAG1, MIC3 and ROP2 genes were amplified from the genomic DNA, with product sizes of 1, 1.1 and 1.7 kb. The eukaryotic expression plasmids pShuttle-SAG1, pLVX-Zsgreen-MIC3 and pLVX-rfp-ROP2 were constructed, and the corresponding fluorescences (blue, green and red) were observed after transfection. On day 28 after mouse vaccination, ELISA showed that the mean A(450) values for serum IgG in groups A, B and C were (0.620±0.029), (0.741±0.040) and (1.561±0.131), respectively, with the group C value being significantly higher than the others (P<0.01). CONCLUSION: The cocktail DNA vaccine comprising T. gondii SAG1, MIC3 and ROP2 shows promising immunogenicity in mice, and the fluorescent protein expression vectors are reliable tools for expression of the target genes.
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Vacunas Antiprotozoos/inmunología , Toxoplasma , Toxoplasmosis/prevención & control , Vacunas de ADN/inmunología , Animales , Anticuerpos Antiprotozoarios/sangre , Antígenos de Protozoos/inmunología , Vectores Genéticos , Proteínas de la Membrana/inmunología , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Plásmidos , Proteínas Protozoarias/inmunologíaRESUMEN
BACKGROUND: Awake fibreoptic intubation (AFOI) frequently requires sedation, anxiolysis and relief of discomfort without impairing ventilation and depressing cardiovascular function. The goal is to allow the patient to be responsive and co-operative. Medications such as fentanyl, remifentanil, midazolam and propofol have been reported to assist AFOI; however,these agents are associated with cardiovascular or respiratory adverse effects. Dexmedetomidine has been proposed as an alternative to facilitate AFOI. OBJECTIVES: The primary objective of this review is to evaluate and compare the efficacy and safety of dexmedetomidine in the management of patients with a difficult or unstable airway undergoing awake fibreoptic intubation (AFOI). SEARCH METHODS: We searched the Cochrane Central Register of Controlled Trials (CENTRAL; 2012, Issue 5), MEDLINE (1966 to May 2012) through Ovid, EMBASE (1980 to May 2012) and Web of Science (1945 to May 2012); we screened the reference lists of all eligible trials and reviews to look for further trials and contacted authors of trials to ask for additional information. We searched for ongoing trials at http://www.controlledtrials.com/ and http://clinicaltrials.gov/ . We reran our search of all databases listed above on 21 November 2013. SELECTION CRITERIA: We included published and unpublished randomized controlled trials, regardless of blinding or language of publication, in participants 18 years of age or older who were scheduled for an elective AFOI because of an anticipated difficult airway. Participants received dexmedetomidine or control medications. DATA COLLECTION AND ANALYSIS: Three review authors independently extracted data on study design, participants, interventions and outcomes. We assessed risk of bias using The Cochrane Collaboration's tool. We estimated risk ratios (RRs) or mean differences (MDs) with 95% confidence internals (CIs) for outcomes with sufficient data; for other outcomes, we performed a qualitative analysis. MAIN RESULTS: We identified four randomized controlled trials (RCTs), which included 211 participants. The four trials compared dexmedetomidine with midazolam, fentanyl, propofol or a sodium chloride placebo, respectively. The trials showed low or unclear risk of bias primarily because information provided on allocation concealment and other potential sources of bias was inadequate. Owing to clinical heterogeneity and potential methodological heterogeneity, it was impossible to conduct a full meta-analysis. We described findings from individual studies or presented them in tabular form. Limited evidence was available for assessment of the outcomes of interest for this review. Results of the limited included trials showed that dexmedetomidine significantly reduced participants' discomfort with no significant differences in airway obstruction, low oxygen levels or treatment-emergent cardiovascular adverse events noted during AFOI compared with control groups. When the search was rerun (from May 2012 to November 2013), it was noted that four studies are awaiting assessment. We will deal with these studies when we update the review. AUTHORS' CONCLUSIONS: Small, limited trials provide weak evidence to support dexmedetomidine as an option for patients with an anticipated difficult airway who undergo AFOI. The findings of this review should be further corroborated by additional controlled investigations.
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Agonistas de Receptores Adrenérgicos alfa 2 , Dexmedetomidina , Hipnóticos y Sedantes , Intubación Intratraqueal/métodos , Vigilia , Ansiolíticos , Fentanilo , Humanos , Midazolam , Propofol , Ensayos Clínicos Controlados Aleatorios como AsuntoRESUMEN
MicroRNAs (miRNAs) regulate gene expression by inhibiting translation or targeting messenger RNA (mRNA) for degradation in a posttranscriptional fashion. In this study, we show that ectopic expression of miR-34a-5p reduces the mRNA and protein levels of Krüppel-like factor 4 (KLF4). We also demonstrate that miR-34a targets the 3'-untranslated mRNA region of KLF4 and show that overexpression of miR-34a induces a significant level of apoptosis in BNL CL.2 cells exposed to doxorubicin or 10 Gy X-ray. Our data suggest that the effects of miR-34a on apoptosis occur due to the downregulation of KLF4.
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Factores de Transcripción de Tipo Kruppel/biosíntesis , Hígado/metabolismo , MicroARNs/genética , ARN Mensajero/biosíntesis , Apoptosis/genética , Regulación de la Expresión Génica , Hepatocitos/metabolismo , Humanos , Factor 4 Similar a Kruppel , Factores de Transcripción de Tipo Kruppel/genética , Hígado/citología , ARN Mensajero/genéticaRESUMEN
OBJECTIVE: To clone and express the conservative region of gene encoding tyrosine kinase 4 of Schistosoma japonicum and identify the difference in gene expression between genders of S. japonicum. METHODS: The gene fragment was amplified by reverse transcriptase-polymerase chain reaction (RT-PCR) using the total RNA isolated from adult S. japonicum (Chinese strain) with primers designed according to SmTK4 encoding tyrosine kinase 4. The purified PCR product was ligated with pET28a and the recombinant protein was induced to express, and analyzed by SDS-PAGE, Western blotting and tools of bio-informatics. Subsquently, total RNA was respectively isolated from adult males, females and both worms of S. japonicum. The real-time PCR was performed with corresponding primers after reverse transcription to show the expression levels of the gene in both genders. RESULTS: A 582 bp in size of the DNA fragment was acquired by RT-PCR. Sequence analysis indicated that the fragment showed 91% in homology to that of SmTK4, and the deduced amino acid sequence showed to be 98% identical with that encoded by SmTK4. SDS-PAGE analysis revealed that the relative molecular weight (M(r)) of expressed protein rSjTK4 was approximately 26000. The bio-information analysis demonstrated that the protein had multiple sites of enzymatic activities. The relative number of copies of SjTK4 in male worms was 0.61 +/- 0.29, while 0.03 +/- 0.02 in female worms, showing that the mRNA level of TK4 in male worms was 18 times higher than that in females. CONCLUSION: The conservative region of gene encoding tyrosine kinase 4 of S. japonicum is successfully cloned and expressed. The mRNA level of TK4 in male worms is significantly higher than that in females.
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Proteínas Tirosina Quinasas/genética , Schistosoma japonicum/enzimología , Secuencia de Aminoácidos , Animales , Western Blotting , Clonación Molecular , ADN Complementario , Electroforesis en Gel de Poliacrilamida , Femenino , Expresión Génica , Masculino , Reacción en Cadena en Tiempo Real de la Polimerasa , Proteínas Recombinantes , Schistosoma japonicum/genéticaRESUMEN
OBJECTIVE: To explore the toll-like receptor 7 knocked out (TLR7-/-) mice immune response against Schistosoma japonicum. METHODS: C57BL/6 mice (WT) and TLR7-/- mice (TLR7-/-) were infected with 20 S. japonicum cercariae via shaved abdomen. There were nine mice in each group. At 6 weeks post-infection, mice were sacrificed. Adult worms were harvested by perfusion of the portal venous system, and the number of adult worms was determined. At the time of perfusion, livers were collected, weighed, and digested overnight with 5% potassium hydroxide, and eggs were counted. In addition, spleens were aseptically harvested when WT and TLR7-/- mice were sacrificed at day zero and 6 weeks after S. japonicum infection. After 72 hours of the co-culture with or without S. japonicum eggs, the culture supernatants were collected for cytokine assays by ELISA assay. RESULTS: At 6 weeks after infection, there was no significant difference in number of worms [(10.5 +/- 3.3) vs (9.8 +/- 5.2)] and eggs per gram of liver tissue [(38 251.9 +/- 4 891.5) vs (38 160.9 +/- 3 341.0)] between WT and TLR7-/- mice. As for Th1/Th2 cytokine secretion from spleen cells, the levels of TNF-alpha [(43.7 +/- 9.8) pg/ml] and INF-gamma [(215.2 +/- 35.4) pg/ml] from TLR7-/- infected mice were lower than those of WT infected mice[(63.4 +/- 22.9) pg/ml, (383.5 +/- 253.3) pg/ml]. For Th2 cytokines detection, the production of IL-10 [(1702.6 +/- 572.3) pg/ml] and IL-4 [(59.5 +/- 10.1) pg/ml] from TLR7-/- mice were higher than those of WT mice [(595.2 +/- 386.3) pg/ml, (8.3 +/- 0.9) pg/ml] (P < 0.05, P < 0.01), while IL-4 level [(63.9 +/- 33.9) pg/ml] from TLR7-/- infected mice was higher than those of WT infected mice [(23.3 +/- 11.5) pg/ml]. CONCLUSION: TLR7-/- mice has a dominant Th2 response under the normal state. The absence of TLR7 does not influence the immune response against S. japonicum infection at 6 weeks post-infection.
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Glicoproteínas de Membrana/inmunología , Schistosoma japonicum/inmunología , Esquistosomiasis Japónica/inmunología , Receptor Toll-Like 7/inmunología , Animales , Cercarias , Técnicas de Cocultivo , Citocinas , Ensayo de Inmunoadsorción Enzimática , Hígado , Glicoproteínas de Membrana/deficiencia , Ratones , Ratones Endogámicos C57BL , Bazo , Receptor Toll-Like 7/deficiencia , Factor de Necrosis Tumoral alfaRESUMEN
OBJECTIVE: To determine the accumulation of CD11b+ Gr-1+ myeloid-derived suppressor cells (MDSC) in Schistosorna japonicum-infected mice. METHODS: Twenty-four C57BL/6 mice were infected cutaneously with S. japonicum cercariae. Peripheral blood samples were collected at 1, 2, 6 and 8 weeks post-infection (6 mice for each group). At 6 and 8 weeks post-infection, spleens were removed and a single-cell suspension was prepared. At the same time, 6 healthy mice each served as control. During the different stages of infection, the levels of MDSC, Gr-1+ cells, CD11b+ cells in murine peripheral blood and spleen were detected by flow cytometry. The possible function of MDSC on T cells was evaluated by using a CCK-8 method and CFSE proliferation assay. RESULTS: At 6 and 8 weeks post-infection, the levels of MDSC (38.2%-57.8% and 47.1-77.6%, respectively), Gr-1+ cells (28.9%-44.6%, 40.4%-72.9%), and CD11b+ cells (36.0%-48.1%, 40.3%-68.3%) in infection group were significantly higher than that of the controls (15.1%-20.4%, 8.4%-17.3%, 9.8%-22.6%), and that of infection group at 1 week (16.2%-19.8%, 13.0%-16.8%, 17.6%-19.4%) and 2 weeks (19.8%-29.5%, 17.2%-22.2%, 20.9%-33.3%) post-infection (P < 0.01). No significant difference was found in the levels of MDSC, Gr-1+ cells, CD11b+ cells among infection group at 1 and 2 weeks post-infection and control group. Moreover, the fluctuation trends of these cells in the spleens of infected mice were similar to those cells in peripheral blood (P > 0.05). Strikingly, the proliferation index of normal CD4 T cells was significantly lower after co-culture with Gr-1+ cells isolated from infected mice. CONCLUSION: Schistosoma japonicum infection induces higher level of MDSC in mice, and Gr-1+ cells isolated from the infected mice can significantly inhibit the proliferation of the normal CD4+ T cells.
Asunto(s)
Linfocitos T CD4-Positivos/inmunología , Esquistosomiasis Japónica/inmunología , Bazo/inmunología , Animales , Técnicas de Cocultivo , Citometría de Flujo , Ratones , Ratones Endogámicos C57BL , Bazo/parasitologíaRESUMEN
Radon and its progeny are confirmed to be type I carcinogenic agents accounting for increased risks in 10% of observed lung cancers globally. However, the underlying carcinogenic mechanisms are largely unknown. In the present study, BEAS2B cells were directly exposed twice to 20,000 Bq/m(3) radon gas for 20 min once (first passage) and subsequently 10 times (fifth passage). The fifth-passage cells were then subcultured for 1 and 20 generations (named Rn5-1 and Rn5-20, respectively). Molecular mechanisms indicative of malignant transformation were assessed by determination of apoptosis, seroresistance, and microRNA (miRNA) expression profiles. The microRNA profiles were used to assess the functional annotations of the target genes. Data indicated an increased seroresistance and colony efficiency on soft agar, and enhanced apoptosis resistance in the Rn5-20 cells with significant differential expressions in some miRNA, including hsa-miR-483-3p, hsa-miR-494, hsa-miR-2115*, hsa-miR-33b, hsa-miR-1246, hsa-miR-3202, hsa-miR-18a, hsa-miR-125b, hsa-miR-17*, and hsa-miR-886-3p. Functional annotation demonstrated that these miRNA target genes were predominantly involved in the regulation of cell proliferation, differentiation, and adhesion during the process of malignant transformation, which is associated with signal pathways such as mitogen-activated protein kinase (MAPK), Int and Wg (Wnt), reactive oxygen species (ROS), nuclear factor κB (NF-κB), and other genes regulating cell cycles.
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Contaminantes Radiactivos del Aire/toxicidad , Carcinógenos Ambientales/toxicidad , Transformación Celular Neoplásica/inducido químicamente , MicroARNs/metabolismo , Radón/toxicidad , Transcriptoma/efectos de los fármacos , Apoptosis/efectos de los fármacos , Línea Celular Transformada , Proliferación Celular/efectos de los fármacos , Transformación Celular Neoplásica/genética , Transformación Celular Neoplásica/metabolismo , Transformación Celular Neoplásica/patología , Perfilación de la Expresión Génica , Humanos , Análisis de Secuencia por Matrices de OligonucleótidosRESUMEN
OBJECTIVES: The primary objective of this review was to evaluate and compare the efficacy and safety of dexmedetomidine hydrochloride with the efficacy and safety of opioids for postoperative management of children after tonsillectomy and adenoidectomy. METHODS: We searched the Cochrane Central Register of Controlled Trials (Central) in the Cochrane Library (most recent issue), Medline (1966 to date) through Ovid, Embase (1980 to date), and Web of Science (1945 to date). The number of patients who required rescue analgesics (morphine or fentanyl) in the postanesthesia care unit, the number of patients with emergence agitation, the number of patients with postoperative nausea and vomiting, the time to eye-opening in response to verbal stimuli, and the time to extubation were analyzed. RESULTS: We included 5 trials, consisting of 482 patients in total. There were no significant differences in the number of patients who required rescue analgesics in the postanesthesia care unit, the number of patients with emergence agitation, the number of patients with postoperative nausea and vomiting, or the time to extubation between patients who received dexmedetomidine and those who received opioids. Compared with opioids, dexmedetomidine was associated with a significantly decreased time to eye-opening in response to verbal stimuli (mean difference, -2.11 minutes; 95% confidence interval, -3.32 to -0.91 minutes; p = 0.0006). CONCLUSIONS: Intraoperative use of dexmedetomidine was as effective as opioids in preventing postoperative pain and emergence agitation in children who had undergone tonsillectomy and adenoidectomy.
Asunto(s)
Adenoidectomía , Dexmedetomidina/uso terapéutico , Fentanilo/uso terapéutico , Morfina/uso terapéutico , Dolor Postoperatorio/tratamiento farmacológico , Ensayos Clínicos Controlados Aleatorios como Asunto , Tonsilectomía , Analgésicos no Narcóticos/uso terapéutico , Analgésicos Opioides/uso terapéutico , HumanosRESUMEN
OBJECTIVE: Pigs, as hosts of zoonotic Cryptosporidium species/genotypes, are domestic animals with public health significance. The present study was to characterize the infection rate and species/genotype of Cryptosporidium in pre-weaned and post-weaned pigs from Shanghai and Shaoxing, China. METHODS: A total of 208 fecal samples (42 from pre-weaned piglets, and 166 from post-weaned pigs) were examined by nested PCR of the 18S rRNA gene and analyzed by phylogenetic DNA fragment sequencing of secondary PCR products. RESULTS: Infection was detected in 79 samples (19/42 pre-weaned piglets, and 60/166 post-weaned pigs). C. suis (14/79) and Cryptosporidium pig genotype II (65/79) were identified; piglets were more susceptible to the former (13/14) and post-weaned pigs to the latter (59/65). CONCLUSION: Infection of Cryptosporidium spp. in pigs was age-specific; piglets were more susceptible to C. suis while pigs were more susceptible to Cryptosporidium pig genotype II. These findings combined with the isolation of the two Cryptosporidium from water suggest that pigs may be a source of zoonotic Cryptosporidium water pollution. Improvements in pig feeding practices, sewage discharge, feces disposal and field worker protection are therefore important to prevent potential public health problems.
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Criptosporidiosis/veterinaria , Predisposición Genética a la Enfermedad , Genotipo , Enfermedades de los Porcinos/parasitología , Envejecimiento , Animales , China/epidemiología , Criptosporidiosis/epidemiología , Criptosporidiosis/parasitología , Porcinos , Enfermedades de los Porcinos/epidemiología , DesteteRESUMEN
OBJECTIVE: To investigate the clinical efficacy of plasma exchange (PE) with or without prednisone and hydroxychloroquine (HCQ) for the treatment of systemic lupus erythematosus (SLE) during pregnancy. METHODS: The clinical characteristics of 14 pregnant women with SLE admitted to our hospital were retrospectively analyzed, including 7 only treated with prednisone and HCQ (non-PE group) as well as 7 combined PE (PE group). The delivery situations of 14 patients were recorded. Data like erythrocyte sedimentation rate (ESR), urine protein, platelet count, and SLEDAI scores were compared between two groups before treatment and 3, 6, and 12 months after delivery. RESULTS: Three patients in the non-PE group ended in miscarriage while all patients in the PE group were delivered successfully. Eleven successfully delivered fetuses in the two groups were healthy, and the Apgar scores were over 8. The ESR of the PE group was significantly lower than that of the non-PE group at 6 and 12 months after delivery, while there was no statistical difference in ESR between the two groups before treatment and 3 months after delivery. The ESR and urine protein were significantly higher in the non-PE group at months 3, 6, and 12 postpartum. There was a significant decrease in disease activity postpartum in the PE group compared to predelivery disease activity. The change in platelet counts between the two groups significantly increased over time in the PE group, while SLEDAI scores decreased. CONCLUSIONS: The combination of PE and oral prednisone and HCQ is possibly a more effective treatment than oral prednisone and HCQ alone for patients with active SLE during pregnancy. This treatment option reduces pregnancy loss and promotes the patients' postpartum condition to a certain extent.