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BACKGROUND: Cadmium (Cd) is a widespread toxic heavy metal pollutant in agricultural soil, and Cd accumulation in rice grains is a major intake source of Cd for Asian populations that adversely affect human health. However, the molecular mechanism underlying Cd uptake, translocation and accumulation has not been fully understood in rice plants. RESULTS: In this study, a mutant displaying extremely low Cd accumulation (lcd1) in rice plant and grain was generated by EMS mutagenesis from indica rice cultivar 9311 seeds. The candidate SNPs associated with low Cd accumulation phenotype in the lcd1 mutant were identified by MutMap and the transcriptome changes between lcd1 and WT under Cd exposure were analyzed by RNA-seq. The lcd1 mutant had lower Cd uptake and accumulation in rice root and shoot, as well as less growth inhibition compared with WT in the presence of 5 µM Cd. Genetic analysis showed that lcd1 was a single locus recessive mutation. The SNP responsible for low Cd accumulation in the lcd1 mutant located at position 8,887,787 on chromosome 7, corresponding to the seventh exon of OsNRAMP5. This SNP led to a Pro236Leu amino acid substitution in the highly conserved region of OsNRAMP5 in the lcd1 mutant. A total of 1208 genes were differentially expressed between lcd1 and WT roots under Cd exposure, and DEGs were enriched in transmembrane transport process GO term. Increased OsHMA3 expression probably adds to the effect of OsNRAMP5 mutation to account for the significant decreases in Cd accumulation in rice plant and grain of the lcd1 mutant. CONCLUSIONS: An extremely low Cd mutant lcd1 was isolated and identified using MutMap and RNA-seq. A Pro236Leu amino acid substitution in the highly conserved region of OsNRAMP5 is likely responsible for low Cd accumulation in the lcd1 mutant. This work provides more insight into the mechanism of Cd uptake and accumulation in rice, and will be helpful for developing low Cd accumulation rice by marker-assisted breeding.
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Cádmio/metabolismo , Proteínas de Transporte de Cátions/genética , Oryza/genética , Proteínas de Plantas/genética , Poluentes do Solo/metabolismo , Sequência de Aminoácidos , Transporte Biológico , Proteínas de Transporte de Cátions/química , Proteínas de Transporte de Cátions/metabolismo , Perfilação da Expressão Gênica , Oryza/metabolismo , Filogenia , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Alinhamento de SequênciaRESUMO
Lysophospholipase I (LYPLA1) is an important protein with multiple functions. In this study, the full-length cDNA of the LYPLA1 gene from Ovis aries (OaLypla1) was cloned using primers and rapid amplification of cDNA ends (RACE) technology. The full-length OaLypla1 was 2457 bp with a 5'-untranslated region (UTR) of 24 bp, a 3'-UTR of 1740 bp with a poly (A) tail, and an open reading frame (ORF) of 693 bp encoding a protein of 230 amino acid residues with a predicted molecular weight of 24,625.78 Da. Phylogenetic analysis showed that the OaLypla1 protein shared a high amino acid identity with LYPLA1 of Bos taurus. The recombinant OaLypla1 protein was expressed and purified, and its phospholipase activity was identified. Monoclonal antibodies (mAb) against OaLypla1 that bound native OaLypla1 were generated. Real-time PCR analysis revealed that OaLypla1 was constitutively expressed in the liver, spleen, lung, kidney, and white blood cells of sheep, with the highest level in the kidney. Additionally, the mRNA levels of OaLypla1 in the buffy coats of sheep challenged with virulent or avirulent Brucella strains were down-regulated compared to untreated sheep. The results suggest that OaLypla1 may have an important physiological role in the host response to bacteria. The function of OaLypla1 in the host response to bacterial infection requires further study in the future.
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Brucella melitensis/genética , DNA Complementar/genética , Lisofosfolipase/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Brucella melitensis/enzimologia , Brucelose/imunologia , Brucelose/veterinária , Clonagem Molecular , Feminino , Camundongos Endogâmicos BALB C , Filogenia , Reação em Cadeia da Polimerase , Reação em Cadeia da Polimerase em Tempo Real , Homologia de Sequência de Aminoácidos , OvinosRESUMO
The hydrotalcite with the properties of flame-retardant, eliminating smoke, filling and thermostability is a new kind of inorganic flame retardant. In the work, the MgAl hydrotalcite as flame retardant with Mg/Al molar ratio of 4 (MgAl-LDH) was prepared by using urea as the precipitating agent. The thermolysis behavior of the MgAl-LDH flame retardant was investigated by X-ray diffraction (XRD), fourier transform infrared spectroscopy (FT-IR) and thermogravimetry-differential scanning calorimetry (TG-DSC) as well as self deconvolution and curve-fitting analyses. Thermal phase transition of the MgAl-LDH was clarified, especially the characteristics of the hydroxyl groups (-OH) in the brucite-like layers and the changes in coordinate of the carbonate (CO3(2-)) from the interlayers. Based on thermodynamic data, thermal decomposition process was discussed. By.XRD analysis; it was found that the phase change took place when the decomposition temperature increased. The MgAl-LDH was decarbonated basically to MgAl mixed metal oxides (Mg-Al-O) at 500 °C, and impurity MgAl204 phase formed at 600 °C. According to the analyses of FT-IR, TG-DSC and curve-fitting technique, the hydroxyl groups (-OH) in the brucite-like layers possessed three the ligands such as [Al-OH-Al], [Al-OH-Mg] and [Mg-OH-Mg] modes. Dehydroxylation of the brucite-like layers based on the binding forces, where the [Mg-OH-Mg] among the three modes was the most difficult to be re- moved during the pyrolysis process. In the same way, the CO3(2-) ligands also possessed three modes such as H2O-bridged CO3(2-), monodentate and bidentate coordination modes. Based on the thermodynamic analysis, the thermodynamic properties of the hydrotalcite as flame retardant were evaluated, and the expressions of the Gibbs free energy, (ΔrGθT), as a function of temperature, were derived for the Mg8Al2 (OH)20CO3 crystal. Thermodynamic analysis showed that the removal of -OH from the brucite-like layers was spontaneous process, when the Gibbs free energy (ΔrGθT) was under zero at the temperature (T) above 228. 65 °C. The result and datum were close to the experimental result from the TG-DSC analyses, indicating that the relationship between the Gibbs free energy (ΔrGθT) and temperature (T) from thermodynamic analysis was reliable.
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PURPOSE: PHBHHx (poly(3-hydroxybutyrate-co-3-hydroxyhexanoate)) is an excellent biomaterial for tissue repair. Here, we aim to develop a PHBHHx-based three-dimensional (3D) scaffold system for sustained delivery of proteins (insulin serves as a model protein). METHODS: The insulin-phospholipid complex (INS-PLC) was prepared to enhance the insulin lipophilicity. INS-PLC loaded PHBHHx 3D scaffolds (INS-PLC-SCAs) containing PEG-2000 were fabricated by lyophilization. In vitro release was performed in the medium with or without lipase. The bioactivity of INS-PLC-SCAs was measured in diabetic rats. RESULTS: In vitro release shows that the release rate of INS-PLC-SCAs was very slow (~6% of total insulin was released within 120 days), and PEG-2000 or lipase had no effect on its release pattern. The bioactivity test shows that the hypoglycaemic effect of insulin was maintained after formulated into scaffolds. After subcutaneous (s.c.) implantation, its therapeutic effect lasted for over 130 h, and its bioavailability was enhanced by 4-fold. CONCLUSIONS: PHBHHx based 3D scaffold has a great potential for sustained delivery of proteins, especially growth factors. When growth factors are incorporated, it can serve as a bifunctional system that provides a porous skeleton for cells attachment and proliferation, as well as a matrix for long term release of the loaded growth factors.
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Ácido 3-Hidroxibutírico/química , Caproatos/química , Preparações de Ação Retardada/química , Diabetes Mellitus Experimental/tratamento farmacológico , Hipoglicemiantes/administração & dosagem , Insulina/administração & dosagem , Fosfolipídeos/química , Ácido 3-Hidroxibutírico/metabolismo , Animais , Materiais Biocompatíveis/química , Materiais Biocompatíveis/metabolismo , Caproatos/metabolismo , Preparações de Ação Retardada/metabolismo , Hipoglicemiantes/uso terapêutico , Insulina/uso terapêutico , Masculino , Fosfolipídeos/metabolismo , Ratos , Ratos Sprague-Dawley , Alicerces Teciduais/químicaRESUMO
Objectives: This study aimed to identify the key proteins in the bone mass of ovariectomized (OVX) rats after a period of regular moderate-intensity treadmill exercise and to investigate their effects using tag mass spectrometry and quantitative proteomics with a view to improving the understanding and treatment of postmenopausal osteoporosis. Methods: Sixty three-month-old female Sprague-Dawley tats of specific-pathogen-free grade were randomly and equally divided into a sham operation group, ovariectomized group (OVX) and ovariectomized combined exercise (OVX + EX) group, and the latter took moderate-intensity treadmill exercise for 17 weeks. After this period of time, body composition and bone density were measured using dual-energy x-ray absorptiometry, and serum bone metabolism indicators were measured using an enzyme immunoassay. In addition, the bone microstructure was examined using micro-computed tomography and scanning of the femur, and femur proteins were subject to proteomic analysis. Results: Compared with the rats in the OVX group, the bone metabolism indicators in the OVX + EX group decreased significantly, femur bone density increased significantly, the number of the trabeculae increased, and continuity was higher. In the OVX + EX group, 17 proteins were significantly upregulated and 33 significantly downregulated. The main gene ontology and signaling pathways enriched by the proteins were identified as the tumor necrosis factor-mediated signaling pathways. The protein-protein interaction network identified the key proteins, and the correlation analysis of these proteins and the bone parameters found histone deacetylase 8(HDAC8) and leucine-rich transmembrane and O-methyltransferase domain containing (LRTOMT) and trimethylguanosine synthase 1(TGS1) and ankyrin repeat domain 46(ANKRD46) to be the key targets of exercise in relation to postmenopausal osteoporosis. Conclusion: Moderate-intensity treadmill exercise significantly improved the bone mass of OVX rats, and differentially expressed proteins, such as HDAC8 and LRTOMT and TGS1 and ANKRD46, could be the target of moderate-intensity treadmill exercise.
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Objective: Using RNA-sequencing technology to screen the effect of moderate-intensity treadmill exercise on the sensitive genes that affect bone mass in the peripheral blood mononuclear cells (PBMCs) of ovariectomized (OVX) rats. Methods: Three-month-old female Sprague-Dawley rats of Specific Pathogen Free (SPF) grade were randomly divided into the sham operation (SHAM) group, OVX group, and OVX combined exercise (OVX + EX) group. The OVX + EX group performed moderate-intensity treadmill exercise for 17 weeks. Then, the body composition and bone mineral density (BMD) were measured, and the bone microstructure of the femur was observed. PBMCs were collected from the abdominal aorta, and the differential genes were analyzed by transcriptome sequencing to further screen sensitive genes. Results: (1) In the OVX group, the body weight and body fat content were significantly higher than in the SHAM group while the muscle content and BMD were significantly lower than the SHAM group. (2) The trabecular bone parameters in the OVX group were significantly lower than in the SHAM group, and they were significantly higher in the OVX + EX group than in the OVX group. When compared with the SHAM group, the microstructure of the distal femur trabecular in the OVX group was severely damaged, suggest that the morphological structure of trabecular bone is severely damaged, the number of trabecular bones is reduced, and the thickness becomes thinner, which lead to the widening of the trabecular bone space and the appearance of osteoporosis. The number and continuity of the trabecular bones were higher in the OVX + EX group than in the OVX group. (3) A Venn diagram showed that there were 58 common differential genes, and the differential genes were mainly enriched in the PI3K-Akt signaling pathway. Five sensitive genes were screened including CCL2, Nos3, Tgfb3, ITGb4, and LpL. The expression of CCL2, Nos3, and Tgfb3 genes was closely related to multiple bone parameters. Conclusion: Moderate-intensity treadmill exercise may improve the body composition and bone mass of the OVX group by upregulating CCL2 and other genes of the PBMC. The PBMCs in the peripheral blood can be a useful tool for monitoring the effect of exercise on bone health in postmenopausal osteoporosis.
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The present study aimed to investigate the effect of bezafibrate on glucolipid abnormalities induced by antipsychotics in schizophrenia. Patients in the treatment group (group A) were treated with antipsychotics and a daily dose of 200â¯mg bezafibrate for 12 weeks, and patients in the control group (group B) were treated with antipsychotics; sugar, fat and weight changes before and after the treatment were compared between the two groups. Before treatment the differences in TG, TC, LDL-C, HDL-C, body weight and blood glucose between groups A and B were not statistically significant. However, in group B, levels of TG, TC, LDL-C, body weight and blood glucose after treatment showed statistically significant increases, although levels of HDL-C did not register any statistically significant change. By contract, in group A, there were no statistically significant changes in any of the variables measured. Bezafibrate can prevent an increase in sugar, fat and weight gain in treating schizophrenia patients with antipsychotics, and low doses of bezafibrate are safe in the antipsychotic treatment for schizophrenia.
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Antipsicóticos/administração & dosagem , Bezafibrato/administração & dosagem , Glicemia/efeitos dos fármacos , Hipolipemiantes/administração & dosagem , Risperidona/administração & dosagem , Esquizofrenia/tratamento farmacológico , Adolescente , Adulto , Antipsicóticos/efeitos adversos , Glicemia/metabolismo , Peso Corporal/efeitos dos fármacos , Peso Corporal/fisiologia , Quimioterapia Combinada , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Risperidona/efeitos adversos , Esquizofrenia/sangue , Aumento de Peso/efeitos dos fármacos , Aumento de Peso/fisiologia , Adulto JovemRESUMO
INTRODUCTION: Serological diagnosis of brucellosis is still a great challenge due to the infeasibility of discriminating infected animals from vaccinated ones, so it is necessary to search for diagnostic biomarkers for differential diagnosis of brucellosis. MATERIAL AND METHODS: Cell division cycle 42 (Cdc42) from sheep (Ovis aries) (OaCdc42) was cloned by rapid amplification of cDNA ends (RACE), and then tissue distribution and differential expression levels of OaCdc42 mRNA between infected and vaccinated sheep were analysed by RT-qPCR. RESULTS: The full-length cDNA of OaCdc42 was 1,609 bp containing an open reading frame (ORF) of 576 bp. OaCdc42 mRNAs were detected in the heart, liver, spleen, lung, kidneys, rumen, small intestine, skeletal muscles, and buffy coat, and the highest expression was detected in the small intestine. Compared to the control, the levels of OaCdc42 mRNA from sheep infected with Brucella melitensis or sheep vaccinated with Brucella suis S2 was significantly different (P < 0.01) after 40 and 30 days post-inoculation, respectively. However, the expression of OaCdc42 mRNA was significantly different between vaccinated and infected sheep (P < 0.05 or P < 0.01) on days: 14, 30, and 60 post-inoculation, whereas no significant difference (P > 0.05) was noted 40 days post-inoculation. Moreover, the expression of OaCdc42 from both infected and vaccinated sheep showed irregularity. CONCLUSION: OaCdc42 is not a good potential diagnostic biomarker for differential diagnosis of brucellosis in sheep.
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OBJECTIVE: To investigate beta-sitosterol's inhibitory effects on SiHa cells' growth, and the effects on microtubular system in SiHa cell. METHODS: Proliferation inhibition of SiHa cell line was evaluated by MTT assay. Cell cycle of SiHa cells treated with beta-sitosterol was analyzed by flow cytometry. The expression and distribution of microtubule and microtubule associated protein 2 in SiHa cells were investigated by confocal microscopy. Immunoblotting analysis was used to determine tubulin alpha, microtubule associated protein 2, and the proportion of polymerization of tubulin. RESULTS: beta-sitosterol could obviously inhibit the proliferation of SiHa cells, and induce the accumulation of cells in S phase (rather than the G2/M phase) and mitotic arrest in the cell cycle. Confocal microscopy showed an abnormal microtubular network in SiHa cell treated with beta-sitosterol for 5 days, and the expression of microtubule associated protein 2 was marked down-regulated. Further analysis by immunoblotting confirmed the down-regulation of beta-sitosterol on the expression for both microtubule associated protein 2 and tubulin alpha. Moreover, beta-sitosterol reduced the proportion of polymerization of microtubule in a time-dependent manner. CONCLUSION: beta-sitosterol could down-regulate the expression of tubulin alpha and microtubule associated protein 2 in SiHa cells, and inhibit the microtubular polymerization. Our results suggested an anti-microtubule characteristic of beta-sitosterol which might contribute to the proliferation inhibition of SiHa cells.
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Microtúbulos/efeitos dos fármacos , Microtúbulos/metabolismo , Sitosteroides/farmacologia , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Feminino , Humanos , Immunoblotting , Microscopia Confocal , Proteínas Associadas aos Microtúbulos/biossíntese , Tubulina (Proteína)/biossíntese , Neoplasias do Colo do Útero/metabolismo , Neoplasias do Colo do Útero/patologiaRESUMO
AIM: To study the changes of endogenous interleukin 18 (IL-18) levels and evaluate the role of IL-18 on lung injury following gut ischemia/reperfusion. METHODS: A superior mesenteric artery occlusion model was selected for this research. The mice were randomly divided into four groups: Sham operation (sham), ischemia (0.5 h) followed by different times of reperfusion (I/R), and I/R pretreated with exogenous IL-18 (I/R+IL-18) or IL-18 neutralizing antibody (I/R+IL-18Ab) 15 min before ischemia. Serum IL-18 levels were detected by Western blot and ELISA, and the levels of IL-18 in lung tissue were evaluated by immunohistochemical staining. For the study of pulmonary inflammation, the lung myeloperoxidase (MPO) contents and morphological changes were evaluated. RESULTS: Gut ischemia/reperfusion induced rapid increase of serum IL-18 levels, peaked at 1 h after reperfusion and then declined. The levels of IL-18 in lung tissue were gradually enhanced as the progress of reperfusion. Compared with I/R group, exogenous administration of IL-18 (I/R+IL-18) further remarkably enhanced the pulmonary MPO activity and inflammatory cell infiltration, and in I/R+IL-18Ab group, the content of MPO were significantly reduced and lung inflammation was also decreased. CONCLUSION: Gut ischemia/reperfusion induces the increase of IL-18 expression, which may make IL-18 act as an important proinflammatory cytokine and contribute to gut ischemia/reperfusion-induced lung inflammation.
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Interleucina-18/imunologia , Pneumonia/imunologia , Traumatismo por Reperfusão/imunologia , Doença Aguda , Animais , Intestinos/irrigação sanguínea , Artéria Mesentérica Superior , Camundongos , Camundongos Endogâmicos C57BL , Pneumonia/etiologia , Traumatismo por Reperfusão/complicaçõesRESUMO
Peroxiredoxin 6 (Prdx6), an important antioxidant enzyme that can eliminate reactive oxygen species (ROS) to maintain homeostasis, is a bifunctional protein that possesses the activities of both glutathione peroxidase and phospholipase A2. In this study, a novel full-length Prdx6 cDNA (OaPrdx6) was cloned from Sheep (Ovis aries) using rapid amplification of cDNA ends (RACE). The full-length cDNA of OaPrdx6 was 1753bp containing a 5'-untranslated region (UTR) of 93bp, a 3'-UTR of 985bp with a poly(A) tail, and an open reading frame (ORF) of 675bp encoding a protein of 224 amino acid residues with a predicted molecular weight of 25.07kDa. The recombinant protein OaPrdx6 was expressed and purified, and its DNA protection activity was identified. In order to analyze the Prdx6 protein expression in tissues from O. aries, monoclonal antibodies against OaPrdx6 were prepared. Western blotting results indicated that OaPrdx6 protein could be detected in heart, liver, spleen, lung, kidney, stomach, intestine, muscle, lymph node and white blood cells, and the highest expression was found in lung while the lowest expression in muscle. Compared to the normal sheep group, the mRNA transcription level of Prdx6 in buffy coat was up-regulated in the group infected with a virulent field strain of Brucella melitensis, and down-regulated in the group inoculated with a vaccine strain S2 of brucellosis. The results indicated that Prdx6 was likely to be involved in the host immune responses against Brucella infection, and probably regarded as a molecular biomarker for distinguishing between animals infected with virulent Brucella infection and those inoculated with vaccine against brucellosis.
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DNA Complementar/genética , Peroxirredoxina VI/genética , Sequência de Aminoácidos , Animais , Anticorpos Monoclonais/imunologia , Sequência de Bases , Brucella melitensis/imunologia , Brucelose/imunologia , Brucelose/veterinária , Clonagem Molecular , DNA Complementar/química , Dados de Sequência Molecular , Peroxirredoxina VI/imunologia , Proteínas Recombinantes/imunologia , OvinosRESUMO
Mitochondrial fragmentation plays an important role in the progression of cardiac diseases, such as myocardial infarction and heart failure. Mitochondrial network is controlled by many factors in different cell types. Here we show that the interplay between E2F1, miR-421 and Pink1 regulates mitochondrial morphology and cardiomyocyte cell death. Pink1 reduces mitochondrial fragmentation and protects cardiomyocyte from apoptosis. On the other hand, miR-421 promotes cardiomyocyte mitochondrial fragmentation, apoptosis and myocardial infarction by suppressing Pink1 translation. Finally, we show that transcription factor E2F1 activates miR-421 expression. Knocking down E2F1 suppresses mitochondrial fragmentation, apoptosis and myocardial infarction by affecting miR-421 levels. Collectively, these data identify the E2F1/miR-421/Pink axis as a regulator of mitochondrial fragmentation and cardiomyocyte apoptosis, and suggest potential therapeutic targets in treatment of cardiac diseases.
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Fator de Transcrição E2F1/genética , MicroRNAs/genética , Mitocôndrias Cardíacas/metabolismo , Infarto do Miocárdio/genética , Traumatismo por Reperfusão Miocárdica/genética , Miócitos Cardíacos/metabolismo , Proteínas Quinases/genética , Trifosfato de Adenosina/metabolismo , Animais , Apoptose , Imunoprecipitação da Cromatina , Fator de Transcrição E2F1/metabolismo , Immunoblotting , Camundongos , Camundongos Knockout , Camundongos Transgênicos , Microscopia Eletrônica , Mitocôndrias Cardíacas/ultraestrutura , Infarto do Miocárdio/metabolismo , Infarto do Miocárdio/patologia , Traumatismo por Reperfusão Miocárdica/metabolismo , Traumatismo por Reperfusão Miocárdica/patologia , Miócitos Cardíacos/ultraestrutura , Proteínas Quinases/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Programmed cell death 10 (PDCD10) is a highly conserved adaptor protein. Its mutations result in cerebral cavernous malformations (CCMs). In this study, PDCD10 cDNA from the buffy coat of Small Tail Han sheep (Ovis aries) was cloned from a suppression subtractive hybridization cDNA library, named OaPDCD10. The full-length cDNA of OaPDCD10 was 1343bp with a 639bp open reading frame (ORF) encoding 212 amino acid residues. Tissue distribution of OaPDCD10 mRNA determined that it was ubiquitously expressed in all tested tissue samples, and the highest expression was observed in the heart. The differential expression of OaPDCD10 between infected sheep (challenged with Brucella melitensis) and vaccinated sheep (vaccinated with Brucella suis S2) was also investigated. The results revealed that, compared to the control group, the expression of OaPDCD10 from infected and vaccinated sheep was both significantly up-regulated (p<0.05). Moreover, the expression levels of OaPDCD10 from the vaccinated sheep were significantly higher than the infected sheep (p<0.05) after 30days post-inoculation. The recombinant OaPDCD10 (rOaPDCD10) protein was expressed in Escherichia coli BL21 (DE3), and then purified by affinity chromatography. The rOaPDCD10 protein was demonstrated to induce apoptosis and promote cell proliferation. Our studies are intended to discover potential diagnostic biomarkers of brucellosis to discern infected sheep from vaccinated sheep, and OaPDCD10 could be considered as a potential diagnostic biomarker of brucellosis.
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Proteínas Reguladoras de Apoptose/química , Proteínas Reguladoras de Apoptose/genética , Clonagem Molecular , Carneiro Doméstico/genética , Animais , Apoptose , Proteínas Reguladoras de Apoptose/metabolismo , Brucelose/imunologia , Brucelose/microbiologia , Brucelose/prevenção & controle , Brucelose/veterinária , Proliferação de Células , Escherichia coli/genética , Expressão Gênica , Especificidade de Órgãos , Ovinos , Doenças dos Ovinos/imunologia , Doenças dos Ovinos/microbiologia , Doenças dos Ovinos/prevenção & controleRESUMO
New type of liquid embolic agents based on a liquid crystalline material of glyceryl monooleate (GMO) was developed and evaluated in this study. Ternary phase diagram of GMO, water and ethanol was constructed and three isotropic liquids (ILs, GMO:ethanol:water=49:21:30, 60:20:20 and 72:18:10 (w/w/w)) were selected as potential liquid embolic agents, which could spontaneously form viscous gel cast when contacting with water or physiological fluid. The ILs exhibited excellent microcatheter deliverability due to low viscosity, and were proved to successfully block the saline flow when performed in a device to simulate embolization in vitro. The ILs also showed good cytocompatibility on L929 mouse fibroblast cell line. The embolization of ILs to rabbit kidneys was performed successfully under monitoring of digital subtraction angiography (DSA), and embolic degree was affected by the initial formulation composition and used volume. At 5th week after embolization, DSA and computed tomography (CT) confirmed the renal arteries embolized with IL did not recanalize in follow-up period, and an obvious atrophy of the embolized kidney was observed. Therefore, the GMO-based liquid embolic agents showed feasible and effective to embolize, and potential use in clinical interventional embolization therapy.
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Materiais Biocompatíveis/química , Embolização Terapêutica/métodos , Glicerídeos/química , Cristais Líquidos/química , Animais , Materiais Biocompatíveis/toxicidade , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Etanol/química , Fibroblastos/efeitos dos fármacos , Géis , Glicerídeos/toxicidade , Cristais Líquidos/toxicidade , Camundongos , Transição de Fase , Coelhos , Radiografia , Artéria Renal/diagnóstico por imagem , Artéria Renal/efeitos dos fármacos , Viscosidade , Água/químicaRESUMO
AIM: To screen specifically expressed genes in the development of nerve, muscle, and body axis of amphioxus, Branchiostoma belcheri tsingtauenese. METHODS: A subtractive cDNA library was constructed from the 12-hour amphioxus neurula cDNA after subtractively hybridized with the 6-hour amphioxus gastrula cDNA. The total RNA was extracted from the 12-hour neurula and 6-hour gastrula, then reverse transcribed into cDNA. The 12-hour neurula cDNA was designated as the experimental group (the tester) and the 6-hour gastrula cDNA as the control group (the driver). The differentially expressed sequences were exponentially amplified using suppression PCR. Background was subtracted and differentially expressed sequences were further enriched. The PCR products were ligated to the T Vector. After transformation of the recombinant plasmid carrying inserted amphioxus cDNA into E.coli host cells, the cDNA library was constructed successfully. RESULTS: Two hundred randomly chosen positive clones were sequenced and some of neurula-specifically expressed genes were obtained. CONCLUSION: SSH is an effective method for searching differentially expressed genes. The subtractive cDNA library we generated provides a tool for further study of regulatory mechanisms of amphioxus early embryonic development.
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Peixes/embriologia , Peixes/genética , Biblioteca Gênica , Neurulação/genética , Animais , Clonagem Molecular , Desoxirribonucleases de Sítio Específico do Tipo II/metabolismo , Plasmídeos/genética , RNA/genéticaRESUMO
Interest in atmospheric dry deposition results mostly from concerns about the effects of the deposited trace elements entering waterbody, soil and vegetation as well as their subsequent health effects. A microwave assisted digestion method followed by inductively coupled plasma mass spectrometric (MAD-ICP/MS) analysis was developed to determine the concentrations of a large number of trace metals in atmospheric dry deposition samples with a heavy matrix of polyurethane foam (PUF). A combination of HNO3-H2O2-HF was used for digestion. The experimental protocol for the microwave assisted digestion was established using two different SRMs (GBW 07401, Soil and GBW 08401, Coal fly ash). Subsequently, blanks and limits of detection for total trace metal concentrations were determined for PUF filter which was used for dry deposition sampling. Finally, the optimized digestion method was applied to real world atmospheric dry deposition samples collected at 10 sites in Jingjinji area in winter from Dec. 2007 to Feb. 2008. The results showed that the area-averaged total mass fluxes ranged between 85 and 912 mg x (m2 x d)(-1), and fluxes of most elements were highest at Baoding and lowest at Xinglong. In addition, the elemental fluxes in urban areas of Beijing, Tianjin and Tangshan were measured to be higher than that in suburb and rural sites. The average fluxes of crust elements (A1, Fe, Mn, K, Na, Ca and Mg) were one to three orders of magnitude higher than anthropogenic elements (Cu, Pb, Cr, Ni, V, Zn and Ba), varying from 151 to 16034 microg x (m2 x d)(-1) versus 14 to 243 microg x (m2 x d)(-1). Zinc was the most abundant heavy metal and calcium the highest of the crust elements while the elements Mo, Co, Cd, As and Be deposited less or even could not be detected. The anthropogenic and crustal contributions were estimated by employing enrichment factors (EF) calculated relative to the average crustal composition. The EF values of all elements except Pb and Zn were below 10, suggesting that local soil and/or dust generally dominate in the dry deposition flux.
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Poluentes Atmosféricos/análise , Espectrometria de Massas/métodos , Metais/análise , Oligoelementos/análise , Ar/análise , Atmosfera/análise , Micro-Ondas , Tamanho da Partícula , Manejo de EspécimesRESUMO
OBJECTIVE: To observe the ultrastructure of human oocytes at different maturity stages and morphologies before and after in vitro maturation. DESIGN: Case report. SETTING: University-based center for human reproduction. PATIENT(S): A 28-year-old patient with polycystic ovary syndrome undergoing IVM/IVF-ET treatment. INTERVENTION(S): Human chorionic gonadotropin was given to the patient 36 hours before her oocytes were retrieved by transvaginal aspiration. MAIN OUTCOME MEASURE(S): Ultrastructure of human oocytes matured in vitro. RESULT(S): Obvious ultrastructural differences were found in oocytes at different maturity stages between those cultured in vitro and those matured in vivo. Degenerate structures were observed in abnormal oocytes. Granulosa cell apoptosis rates from abnormal oocytes were higher than those from normal oocytes. Cumulus cells surrounding morphologically abnormal oocytes had a significantly higher frequency of cell apoptosis, as compared with morphologically normal oocytes. CONCLUSION(S): Obvious ultrastructural differences were found between the oocytes matured in vitro and in vivo.
Assuntos
Fertilização in vitro , Oócitos/fisiologia , Oócitos/ultraestrutura , Síndrome do Ovário Policístico/sangue , Adulto , Transferência Embrionária , Estradiol/sangue , Feminino , Hormônio Foliculoestimulante/sangue , Humanos , Hormônio Luteinizante/sangue , Microvilosidades/ultraestrutura , Gravidez , Zona Pelúcida/ultraestruturaRESUMO
To study the characteristics and sources of particles in Pearl River Delta Region, samples of PM2.5 were collected by high volume sampler, and elements of the samples were determined by ICP-MS. The results showed that concentrations of Pb, V, Cu, As, Zn and Se were 216.24, 15.40, 60.56, 31.81,432.06 and 8.12 ng x m(-3) respectively, those were in high level. Factor analysis on the chemical composition of PM2.5 showed that fuel combustion, metal-working industry, ash and sea salt were the main sources of PM2.5 in this region.
Assuntos
Poluentes Atmosféricos/análise , Exposição Ambiental , Monitoramento Ambiental , Metais/análise , Material Particulado/análise , China , Tamanho da PartículaRESUMO
Samples of aerosol are sampled by high volume sampler from Dec. 2005 to Nov. 2006 at high altitude of 1640 m of Gongga Mountain. Water-soluble ions in PM2.5 and PM10 are analyzed by ion chromatogram(IC). The results shows that the annual concentrations of total water-soluble inorganic ions are respectively 6.46 and 8.86 microg/m3 in PM2.5 and PM10, three major ions of SO4(2-), NO3-; and NH4+, account for respectively 82% and 85% in PM2.5 and PM10, Na+, NH4+, K+, Cl- and SO4(2-) were mostly distributed in fine particles, NO3-, Mg2+ and Ca2+ occupied each one half in coast of crude and fine particles. The correlation of Na+ and Cl- is significantly enhanced in wet season in Gongga Mountain, the correlation coefficient R2 are respectively 0.87 and 0.84 in PM2.5 and PM10. The concentration ratio of SO4(2-) and NH4+ is approximate to 1:2 in PM10, main existence form of SO4(2-) and NH4+ (NH4)2SO4 at Gongga Mountain.
Assuntos
Poluentes Atmosféricos/análise , Poeira/análise , Compostos Inorgânicos/análise , Material Particulado/análise , Aerossóis/análise , Aerossóis/química , Poluentes Atmosféricos/química , Altitude , Atmosfera , China , Monitoramento Ambiental , Nitratos/análise , Tamanho da Partícula , Compostos de Amônio Quaternário/análise , Solubilidade , Ácidos de Enxofre/análiseRESUMO
In order to study the characteristics and sources of atmospheric particles before and in heating period, samples of atmospheric particles were collected in November of 2006. Concentrations of elements in particles were determined with ICP-MS. The results shows that concentrations of As, Se, Mo, Cd in heating period are more than twice of those before heating period, and there is sharp increasing of concentrations of Zn, Pb, Tl, K, Se, As, Cu, Cd, Ag in fine particles in heating period. Furthermore, Zn, Na are associated with finer particles in heating period than those before heating period. Factor analysis on the chemical composition of particles shows that contribution of combustion and biomass burning goes up and contribution of crust goes down in heating period.