RESUMO
PHYTOCHROME INTERACTING FACTORS (PIFs) integrate light and temperature signs to control plant growth and development. However, little is known about PIFs in crop plants such as cotton. Here, we identified 68 PIF proteins and their coding genes from an allotetraploid and three diploid ancestors. Cotton PIFs contain typical ACTIVEPHYA-BINDING (APA) and ACTIVE PHYB-BINDING (APB) motifs by which they bind to phytochrome phyA and phyB, respectively, and have a BASIC HELIX-LOOP-HELIX (bHLH) domain and a nuclear localization sequence necessary for bHLH-type transcription factors. Bioinformatics analysis showed that the promoter of each PIF gene contains multiple cis-acting elements and that the evolution of cotton genomes probably underwent loss, recombination, and tandem replication. Further observations indicated that the sensitivity of cotton PIF expression to high temperature was significantly different from that to low temperature. We found that allotetraploid Gossypium hirsutum PIF4a (GhPIF4a) was induced by high temperature. GhPIF4a promotes flowering in cotton and Arabidopsis and binds to the promoter of GhFT (G. hirsutum FLOWERING LOCUS T), and binding increased with increasing temperature. Our work identifies the evolutionary and structural characteristics and functions of PIF family members in cotton.
Assuntos
Proteínas de Arabidopsis , Arabidopsis , Fitocromo , Fitocromo/genética , Fitocromo/metabolismo , Gossypium/genética , Gossypium/metabolismo , Temperatura , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Regulação da Expressão Gênica de PlantasRESUMO
BACKGROUND: Carotid plaque (CP) formation is an important consequence of atherosclerosis and leads to significant complications. Levels of neuropeptide Y (NPY), which is a sympathetic neurotransmitter, are elevated in cardiovascular diseases. It also has important roles in inflammatory conditions. This study aimed to explore the relationship between serum NPY and CP and to study further the influence of NPY and inflammatory factors on CP. METHODS: This cross-sectional study was conducted among 300 adults who underwent a health examination at the Second Affiliated Hospital of Fujian Medical University in Fujian Province, of whom 177 were finally enrolled. The participants were divided into the CP (n = 120) and non-CP (NCP) or control (n = 57) groups according to the results of carotid artery color Doppler ultrasound. The CP group was further classified into stable plaque (SP, n = 80) and vulnerable plaque (VP, n = 40) groups based on plaque characteristics. Serum NPY and pro-inflammatory cytokine tumor necrosis factor-α (TNF-α) levels were examined. Univariate and correlation analyses were used to evaluate the correlation between serum NPY levels, pro-inflammatory cytokines, and the CP phenotype. RESULTS: The serum NPY and TNF-α levels of patients in the CP group were significantly higher than those in individuals from the NCP group [ (177.30 ± 43.29) pg.mL- 1 vs. (121.53 ± 40.16)pg.mL- 1, P < 0.001; (41.94 ± 14.19) pg.mL- 1 vs.(33.54 ± 13.37)pg.mL- 1, P = 0.003]. The serum NPY levels of the patients in the VP group were significantly higher than those in patients from the SP group [(191.67 ± 39.87)ng.L- 1 vs.(170.12 ± 43.37)ng.L- 1, P = 0.01, P < 0.05]. Serum TNF-α and NPY levels were positively correlated among patients from the CP group (r = 0.184, P = 0.044). The binary logistic regression analysis showed that serum NPY and TNF-α were independent influencing factors of CP [(OR = 1.029, P < 0.001);(OR = 1.030, P = 0.023)]. The area under the ROC curve of NPY predicting the CP showed statistical significance at a value of 0.819. CONCLUSION: Together, elevated serum NPY levels seem to be associated with the occurrence of coronary atherosclerosis in Chinese adults.
Assuntos
Neuropeptídeo Y , Placa Aterosclerótica , Adulto , Humanos , Estudos Transversais , Fator de Necrose Tumoral alfa , Citocinas , Artérias Carótidas , ChinaRESUMO
Vascular calcification is caused by the deposition of calcium salts in the intimal or tunica media layer of the aorta, which increases the risk of cardiovascular events and all-cause mortality. However, the mechanisms underlying vascular calcification are not fully clarified. Recently it has been shown that transcription factor 21 (TCF21) is highly expressed in human and mouse atherosclerotic plaques. In this study we investigated the role of TCF21 in vascular calcification and the underlying mechanisms. In carotid artery atherosclerotic plaques collected from 6 patients, we found that TCF21 expression was upregulated in calcific areas. We further demonstrated TCF21 expression was increased in an in vitro vascular smooth muscle cell (VSMC) osteogenesis model. TCF21 overexpression promoted osteogenic differentiation of VSMC, whereas TCF21 knockdown in VSMC attenuated the calcification. Similar results were observed in ex vivo mouse thoracic aorta rings. Previous reports showed that TCF21 bound to myocardin (MYOCD) to inhibit the transcriptional activity of serum response factor (SRF)-MYOCD complex. We found that SRF overexpression significantly attenuated TCF21-induced VSMC and aortic ring calcification. Overexpression of SRF, but not MYOCD, reversed TCF21-inhibited expression of contractile genes SMA and SM22. More importantly, under high inorganic phosphate (3 mM) condition, SRF overexpression reduced TCF21-induced expression of calcification-related genes (BMP2 and RUNX2) as well as vascular calcification. Moreover, TCF21 overexpression enhanced IL-6 expression and downstream STAT3 activation to facilitate vascular calcification. Both LPS and STAT3 could induce TCF21 expression, suggesting that the inflammation and TCF21 might form a positive feedback loop to amplify the activation of IL-6/STAT3 signaling pathway. On the other hand, TCF21 induced production of inflammatory cytokines IL-1ß and IL-6 in endothelial cells (ECs) to promote VSMC osteogenesis. In EC-specific TCF21 knockout (TCF21ECKO) mice, VD3 and nicotine-induced vascular calcification was significantly reduced. Our results suggest that TCF21 aggravates vascular calcification by activating IL-6/STAT3 signaling and interplay between VSMC and EC, which provides new insights into the pathogenesis of vascular calcification. TCF21 enhances vascular calcification by activating the IL-6-STAT3 signaling pathway. TCF21 inhibition may be a new potential therapeutic strategy for the prevention and treatment of vascular calcification.
Assuntos
Placa Aterosclerótica , Calcificação Vascular , Animais , Humanos , Camundongos , Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Células Cultivadas , Células Endoteliais/metabolismo , Interleucina-6/metabolismo , Músculo Liso Vascular/metabolismo , Miócitos de Músculo Liso/metabolismo , Osteogênese , Placa Aterosclerótica/metabolismo , Transdução de Sinais , Fator de Transcrição STAT3/metabolismo , Calcificação Vascular/genética , Calcificação Vascular/patologiaRESUMO
The present study aims to construct an elderly vitality index evaluation system and develop a comprehensive vitality evaluation scale for the elderly to reasonably evaluate the vitality level of the elderly in China, so as to provide a reference for promoting the realization of "active aging" and "healthy aging". Literature research and in-depth interview were used to collect the senile vitality sensitive indexes. The indexes were screened and corrected by Delphi expert consultation method, item analysis method based on classical test theory, factor analysis method, and reliability and validity analysis method. The analytic hierarchy process was used to calculate the weight of each level of indexes. An elderly vitality evaluation system including 4 first-level indexes and 24 second-level indexes was constructed. The consistency test results of all levels of indicators showed that the consistency index (CI) and consistent ratio (CR) were both less than 0.1, which met the requirements and showed satisfactory consistency. The weights of exercise vitality, nutritional vitality, psychological vitality and social vitality were 0.263, 0.141, 0.455 and 0.141, respectively. In conclusion, the comprehensive vitality scale constructed for the Chinese elderly is reliable and scientific, and can be used to evaluate the vitality of the elderly.
Assuntos
Envelhecimento , Processo de Hierarquia Analítica , Humanos , Idoso , Reprodutibilidade dos Testes , Técnica Delphi , China , Inquéritos e QuestionáriosRESUMO
BACKGROUND: The association between serum total indoxyl sulfate (tIS), and cardiovascular disease (CVD) and all-cause mortality is a matter of debate. In the current study we sought to determine the association, if any, between serum tIS, and all-cause and CVD-associated mortality in patients on maintenance hemodialysis (MHD). METHODS: A prospective cohort study was conducted involving 500 MHD patients at Dalian Municipal Central Hospital from 31 December 2014 to 31 December 2020. Serum tIS levels were measured at baseline and classified as high (≥44.16 ng/ml) or low (< 44.16 ng/ml) according to the "X-tile" program. Besides, the associations between continuous serum tIS and outcomes were also explored. Predictors were tested for colinearity using variance inflation factor analysis. Hazard ratios (HRs) and 95% confidence intervals (CIs) were calculated using Cox proportional hazards regression models. Restricted cubic spline model was performed to assess dose-response relationships between tIS concentration and all-cause and CVD mortality. RESULTS: During a 58-month median follow-up period, 224 deaths (132 CVD deaths) were documented. After adjustment for potential confounders, the serum tIS level was positively associated with all-cause mortality (HR = 1.02, 95% = 1.01-1.03); however, we did not detect a significant association when tIS was a dichotomous variable. Compared with the MHD population with a serum tIS level < 44.16 ng/ml, the adjusted HR for CVD mortality among those with a serum tIS level ≥ 44.16 ng/ml was 1.76 (95% = 1.10-2.82). Furthermore, we also noted the same association when the serum tIS level was a continuous variable. CONCLUSION: The serum tIS level was associated with higher risk of all-cause and CVD mortality among MHD patients. Further prospective large-scale studies are required to confirm this finding.
Assuntos
Doenças Cardiovasculares , Indicã , Humanos , Modelos de Riscos Proporcionais , Estudos Prospectivos , Diálise RenalRESUMO
Two novel strains, designated 92R-1T and 9PBR-1T, were isolated from abandoned lead-zinc ore collected in Meizhou, Guangdong Province, PR China. Phylogenetic analyses based on 16S rRNA gene sequences showed that they fell into the genus of Hymenobacter and formed two distinct lineages. Strain 92R-1T was most closely related to Hymenobacter wooponensis JCM 19491T (98.7â%) and Hymenobacter gelipurpurascens LMG 21873T (98.5â%), while strain 9PBR-1T was most closely related to Hymenobacter chitinivorans LMG 21951T (99.0â%), Hymenobacter elongatus JCM 17223T (98.7â%) and Hymenobacter aquaticus JCM 31653T (98.1â%). Strain 92R-1Tshared average nucleotide identity values of 80.0-83.7â% and digital DNA-DNA hybridization values of 23.1-27.1â% with its closely related type strains, respectively, while strain 9PBR-1T shared corresponding values of 80.3-83.2â% and 23.6-26.7â% with its closely related type strains, respectively. The two novel strains could be clearly distinguished from their closely related type strains by enzyme activities and substrates assimilation, respectively. Both of them took iso-C15:0, summed feature 3 (C16:1 ω7c and/or C16:1 ω6c), summed feature 4 (iso-C17:1 I and/or anteiso-C17:1 B) and C16:1 ω5c as major fatty acids, and showed clear differences from their closely relatives in the contents of several components. They contained menaquinone 7 as the major respiratory quinone and phosphatidylethanolamine as the dominant polar lipid. The G+C contents of strains 92R-1T and 9PBR-1T were 56.7 and 59.5 mol%, respectively. The results clearly supported that strains 92R-1T and 9PBR-1T represent two distinct novel species within the genus Hymenobacter, for which the names Hymenobacter fodinae sp. nov. (type strain 92R-1T=GDMCC 1.1493T=JCM 32697T) and Hymenobacter metallicola sp. nov. (type strain 9PBR-1T=GDMCC 1.1491T=JCM 32698T) are proposed.
Assuntos
Cytophagaceae/classificação , Mineração , Filogenia , Técnicas de Tipagem Bacteriana , Composição de Bases , China , Cytophagaceae/isolamento & purificação , DNA Bacteriano/genética , Ácidos Graxos/química , Chumbo , Hibridização de Ácido Nucleico , Fosfatidiletanolaminas/química , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Vitamina K 2/análogos & derivados , Vitamina K 2/química , ZincoRESUMO
A novel pink-pigmented strain, designated 6HR-1T, was isolated from tungsten mine tailings in Jiangxi Province, PR China. Cells were Gram-stain-negative, aerobic, non-spore-forming, rod-shaped and motile with a polar flagellum (monotrichous). It could not utilize methanol, methylamine, formaldehyde or formate as a sole carbon source. The methanol dehydrogenase mxaF gene was absent but the xoxF gene was present. Phylogenomic and 16S rRNA gene phylogenetic analyses clearly showed that strain 6HR-1T was affiliated to the genus Methylobacterium and closely related to 'Methylobacterium terrae' 17Sr1-28T (98.6 %), Methylobacterium platani JCM 14648T (97.7 %), Methylobacterium variabile DSM 16961T (97.7 %) and Methylobacterium currus KACC 19662T (97.4 %). The average nucleotide identity and digital DNA-DNA hybridization values between strain 6HR-1T and its closely related type species were 87.4-88.7 and 33.2-36.3 %, respectively. It had summed feature 8 (C18 : 1 ω7c and/or C18 : 1 ω6c) as the major fatty acid and ubiquinone 10 as the predominant respiratory quinone. Polyphasic characterization supported that strain 6HR-1T represents a novel species of the genus Methylobacterium, for which the name Methylobacterium nonmethylotrophicum sp. nov. is proposed with the type strain 6HR-1T (=GDMCC 1.662T=KCTC 42760T).
Assuntos
Methylobacterium/classificação , Mineração , Filogenia , Tungstênio , Técnicas de Tipagem Bacteriana , Composição de Bases , China , DNA Bacteriano/genética , Ácidos Graxos/química , Genes Bacterianos , Methylobacterium/isolamento & purificação , Hibridização de Ácido Nucleico , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Ubiquinona/análogos & derivados , Ubiquinona/químicaRESUMO
BACKGROUND: Cryptosporidium and Enterocytozoon bieneusi are two important pathogens with zoonotic potential that cause enteric infections in a wide range of hosts, including humans. Both are transmitted from animals to humans by direct contact or through contaminated equipment. Bears are frequently found in Chinese zoos as ornamental animals as well as farmed as commercial animals, and are therefore in close contact with zoo- or farm-keepers, but the prevalence and zoonotic potential of Cryptosporidium and E. bieneusi in bears is poorly understood. In this study, we aimed to provide data on the occurrence and genetic diversity of Cryptosporidium and E. bieneusi in Asiatic black bears from Heilongjiang and Fujian, China. From May 2015 to December 2017, 218 fresh fecal specimens were collected from captive Asiatic black bears in Heilongjiang (n = 36) and Fujian (n = 182), China. Cryptosporidium and E. bieneusi were examined by PCR amplification of the partial small subunit of ribosomal DNA (SSU rDNA) and the internal transcribed spacer (ITS) region of rDNA, respectively. C. andersoni-positive isolates were subtyped through PCR analysis of the four minisatellite/microsatellite (MS1, MS2, MS3 and MS16) loci. RESULTS: The overall prevalence of Cryptosporidium and E. bieneusi were 2.4% (4/218) and 6.4% (14/218), respectively, with 2.8% (1/36) and 22.2% (8/36) in the Heilongjiang Province, and 1.6% (3/182) and 3.3% (6/182) in the Fujian Province. Sequence analysis confirmed the presence of Cryptosporidium species: C. andersoni (n = 3) and a genotype termed Cryptosporidium rat genotype IV (n = 1). All three identified C. andersoni belonged to the MLST subtype A4, A4, A4, A1. Two known E. bieneusi genotypes D (n = 4) and SC02 (n = 10) were identified, both of which belong to zoonotic Group 1. CONCLUSIONS: This is the first report of C. andersoni and Cryptosporidium rat genotype IV in bears. The discovery of the zoonotic potential of E. bieneusi genotype D in bears highlights its significant zoonotic potential and potential threat to human health.
Assuntos
Criptosporidiose/epidemiologia , Cryptosporidium/isolamento & purificação , Enterocytozoon/isolamento & purificação , Microsporidiose/veterinária , Ursidae/microbiologia , Animais , China/epidemiologia , Cryptosporidium/genética , DNA Ribossômico , Enterocytozoon/genética , Repetições de Microssatélites , Microsporidiose/epidemiologia , Reação em Cadeia da Polimerase/veterinária , Prevalência , Zoonoses/epidemiologiaRESUMO
An aerobic, non-motile, Gram-stain-negative, red-to-pinkish and rod-shaped bacterium, designated 9PBR-2T, was isolated from an abandoned lead-zinc ore sample collected from Meizhou, Guangdong Province, PR China. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain 9PBR-2T belongs to the genus Hymenobacter and was most closely related to Hymenobacter rigui KCTC 12533T (98.0â%), Hymenobacter swuensis KCTC 32018T (97.8â%) and Hymenobacter perfusus LMG 26000T (97.6â%). The calculated average nucleotide identity values based on whole genome sequences between strain 9PBR-2T and closely related type strains ranged from 81.3 to 84.1â%. Correspondingly, the digital DNA-DNA hybridization values ranged from 25.5 to 28.1â%. The major fatty acids of strain 9PBR-2T were iso-C15:0, anteiso-C15:0, C16:1ω5c, summed feature 3 (C16:1ω6c and/or C16:1ω7c) and summed feature 4 (iso-C17:1 I and/or anteiso-C17:1 B). It contained menaquinone 7 (MK-7) as the major isoprenoid quinone and phosphatidylethanolamine as the major polar lipid. The genomic DNA G+C content based on whole genome sequence was 59.8 mol%. Characterization based on phylogenetic, chemotaxonomic and phenotypic analyses clearly indicated that strain 9PBR-2T represents a novel species of the genus Hymenobacter, for which the name Hymenobactermetallilatus sp. nov. is proposed. The type strain is 9PBR-2T (=GDMCC 1.1492T=JCM 32699T).
Assuntos
Cytophagaceae/classificação , Mineração , Filogenia , Técnicas de Tipagem Bacteriana , Composição de Bases , China , Cytophagaceae/isolamento & purificação , DNA Bacteriano/genética , Ácidos Graxos/química , Chumbo , Hibridização de Ácido Nucleico , Fosfatidiletanolaminas/química , Pigmentação , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Vitamina K 2/análogos & derivados , Vitamina K 2/química , ZincoRESUMO
It is widely accepted that live vaccines elicit higher immune protection than inactivated vaccines. However, the mechanisms are largely unknown. Here, an array with 64 recombinant outer membrane proteins of Vibrio parahemolyticus was developed to explore antibody responses of live and inactivated V. parahemolyticus post immunization of the 8th, 12th, 16th and 20th day. Among the 64 outer membrane proteins, 28 elicited antibody generation. They were all detected in live vaccine-induced immunity but only 15 antibodies were found in inactivated vaccine-induced immunity. Passive immunization showed that higher percent survival was detected in live than inactivated vaccine-induced immunities. Active immunization indicated that out of 19 randomly selected outer membrane proteins, 5 stimulated immune protection against V. parahemolyticus infection. Among them, antibodies to VP2309 and VPA0526 were shared in mice immunized by live or inactivated vaccines, whereas antibodies to VPA0548, VPA1745, and VP1667 were only found in mice immunized by live vaccine. In addition, live V. parahemolyticus stimulated earlier antibody response than inactivated bacteria. These results indicate that not all of the outer membrane proteins elicited antibody responses when they work together in the form of live or inactivated bacteria; live vaccine elicits more protective antibodies, which contribute to higher immune protection in live vaccine than inactivated vaccine. Notably, the recombinant proteins might be different from those separated from live bacteria, and they might be different in their immunogenic potencies.
Assuntos
Anticorpos Antibacterianos/biossíntese , Proteínas da Membrana Bacteriana Externa/imunologia , Vacinas Bacterianas/imunologia , Imunidade Humoral/efeitos dos fármacos , Vibrioses/prevenção & controle , Animais , Proteínas da Membrana Bacteriana Externa/administração & dosagem , Proteínas da Membrana Bacteriana Externa/genética , Vacinas Bacterianas/administração & dosagem , Vacinas Bacterianas/genética , Temperatura Alta , Soros Imunes/administração & dosagem , Imunização Passiva/métodos , Imunogenicidade da Vacina , Camundongos , Análise Serial de Proteínas , Proteínas Recombinantes/administração & dosagem , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Análise de Sobrevida , Vacinas de Produtos Inativados , Vibrioses/imunologia , Vibrioses/microbiologia , Vibrioses/mortalidade , Vibrio parahaemolyticus/efeitos dos fármacos , Vibrio parahaemolyticus/crescimento & desenvolvimento , Vibrio parahaemolyticus/imunologia , Peixe-ZebraRESUMO
BACKGROUND: miR-193a has been shown to be involved in a variety of biological processes, including cell proliferation, differentiation, and apoptosis. However, little is known about how miR-193a regulates osteogenic differentiation. METHODS: We employed RT-qPCR to determine the level of miR-193a and mRNA level of HMGB1 and osteoblast-specific markers (Runx2, ALP, OSX, OCN). Besides, westernblot was used to probe protein level of phosphorylated MAPK family members and ß-catenin. Bioinformatic analysis was used to predict the putative binding sequence of miR-193a to the 3'-UTR of HMGB1 and we confirmed this result by dual luciferase reporter assay. Alizarin red staining assay (ARS) and alkaline phosphatase activity (ALP) were performed to detect osteogenic differentiation. RESULTS: miR-193a was downregulated in OM (osteogenic medium)induced hBMSC. More interestingly, we found that miR-193a mimic attenuated matrix mineralization and alkaline phosphatase activity, whereas miR-193a inhibitor exerted the opposite phenotypes. Mechanistically, we observed that miR-193a played an inhibitory role in expression of osteoblast-specific markers and activation of MAPK and Wnt signaling pathways. Bioinformatic analysis and dual luciferase assay demonstrated that miR-193a directly targeted 3'-UTR of HMGB1. Furthermore, we overexpressed HMGB1 in miR-193a overexpressed hBMSC to establish that HMGB1 acted as downstream target of miR-193a-inhibited osteogenic differentiation. CONCLUSIONS: Here, we reveal miR-193a plays a suppressive role in osteogenic differentiation of hBMSC via targeting HMGB1. These findings provide a novel mechanism underlying osteogenic differentiation and offer therapeutical strategy for bone formation.
Assuntos
Proteína HMGB1/genética , Células-Tronco Mesenquimais/citologia , MicroRNAs/genética , Osteoblastos/citologia , Osteogênese , Diferenciação Celular , Células Cultivadas , Regulação para Baixo , Humanos , Sistema de Sinalização das MAP Quinases , Masculino , Células-Tronco Mesenquimais/metabolismo , Osteoblastos/metabolismo , Regulação para Cima , Via de Sinalização WntRESUMO
Polyvalent vaccines against more than one species of pathogens are especially important due to the complex ecosystem in aquaculture. We have previously shown that the development of polyvalent vaccines by shuffling six ompA genes from different bacteria with V. parahaemolyticus VP0764 primers. Here, we used the same 6 genes, V. alginolyticus VA0764 and VA1186, V. parahaemolyticus VP0764 and VP1186, E. tarda ompA and E. coli ompA, but with E. tarda ompA primers to develop new polyvalent vaccines. By this approach, we identified 7 potential polyvalent vaccines that were effective against both V. alginolyticus and E. tarda infections. Furthermore, the innate immunity triggered by the vaccines were also explored in three groups, no protection (group I), protection against V. alginolyticus (group II), and protection against both V. alginolyticus and E. tarda (group III). The transcription of IL-1ß, IL-6, IL-8, C3b and NF-kB were significantly increased in group II and group III but not group I, where the expression level of group III was higher than group II. In addition, differential activities of succinate dehydrogenase were detected among the three groups. These results indicate the expansion of polyvalent vaccine reservoir with the same shuffling genes but different primers, and promote the understanding of the mechanisms of polyvalent vaccines based on vaccine-induced innate immunity.
Assuntos
Proteínas da Membrana Bacteriana Externa/imunologia , Vacinas Bacterianas/imunologia , Doenças dos Peixes/imunologia , Peixe-Zebra/imunologia , Animais , Proteínas da Membrana Bacteriana Externa/administração & dosagem , Vacinas Bacterianas/administração & dosagem , Edwardsiella tarda/fisiologia , Infecções por Enterobacteriaceae/imunologia , Distribuição Aleatória , Vibrioses/imunologia , Vibrio alginolyticus/fisiologiaRESUMO
Vaccines are the most economic, efficient and environment-friendly agents in protecting host against bacterial infection. In aquaculture, polyvalent vaccines targeting more than one bacterial specie are highly demanded due to the presence of various types of bacterial pathogens in farming environment. Here eighteen genes encoding outer membrane proteins of Vibrio parahaemolyticus were cloned and expressed. The expressed recombinant proteins were used for antiserum preparation. Passive and active immune protection of the antiserum and recombinant proteins was investigated in the zebrafish model. Two recombinant proteins, VP1667 and VP2369, showed effective immune protection against at least two genera of bacteria, Vibrio (V. parahaemolyticus and V. alginolyticus), Pseudomonas (P. fluorescens) or/and Aeromonas (A. hydrophila), and thereby are potential polyvalent vaccine candidates to defend against bacterial infection in fish farming. Furthermore, the mechanisms for the two polyvalent vaccines in triggering immune response were explored. Antiserum to VP1667 or VP2369 was not cross-reacted with P. fluorescens and A. hydrophila, whereas both recombinant proteins induced significant innate immune response. Comparatively, VP1667 stimulates stronger lymphokine and monokine, and VP2369 induces stronger humoral immune response, while both produce similar NF-κB, COX-2, TLR-1 and TLR-3 expression. Our results identify two polyvalent vaccines and demonstrate characteristics features of their cross-protection at the content of the innate immune response.
Assuntos
Proteínas da Membrana Bacteriana Externa/imunologia , Vacinas Bacterianas/imunologia , Imunidade Inata , Imunização Passiva/veterinária , Vacinação/veterinária , Vibrio parahaemolyticus/imunologia , Peixe-Zebra/imunologia , Animais , Antígenos de Bactérias/imunologia , Clonagem Molecular , Proteínas Recombinantes/imunologiaRESUMO
Our previous studies demonstrated that molecular breeding via DNA shuffling directs the evolution of polyvalent vaccines with desired traits, which leads to generation of polyvalent ompA vaccines using Vibrio alginolyticus VA0764 primers. Here, we replaced VA0764 primers with Edwardsiella tarda ompA primers to generate new polyvalent ompA vaccines by DNA shuffling of the same five ompA genes from four species of bacteria E. tarda, V. parahaemolyticus, V. alginolyticus and Escherichia coli. We identified four polyvalent vaccine candidates from a eukaryotic expressing library EompAs-FE containing 82 ompAs using active immune protection against V. alginolyticus and E. tarda. Furthermore, we explored mechanisms of polyvalent vaccine candidates by investigation of the innate immune response to these ompAs, and found that expression of IL-1ß, IL-8, IL-15, COX-2, IFN-γ, TLR-1, TLR-3 and C3b genes was elevated as a characteristic feature of these polyvalent vaccine candidates. These results indicate that use of different primers to construct a DNA library selects new evolution of polyvalent vaccines with desired traits, and polyvalent ompA vaccines elicit high innate immune response.
Assuntos
Proteínas da Membrana Bacteriana Externa/imunologia , Vacinas Bacterianas/imunologia , Edwardsiella tarda/imunologia , Doenças dos Peixes/imunologia , Vibrio alginolyticus/imunologia , Peixe-Zebra , Animais , Proteínas da Membrana Bacteriana Externa/genética , Proteínas de Bactérias/genética , Proteínas de Bactérias/imunologia , Embaralhamento de DNA/veterinária , Edwardsiella tarda/genética , Infecções por Enterobacteriaceae/imunologia , Infecções por Enterobacteriaceae/microbiologia , Infecções por Enterobacteriaceae/veterinária , Escherichia coli/genética , Escherichia coli/imunologia , Proteínas de Escherichia coli/genética , Proteínas de Escherichia coli/imunologia , Doenças dos Peixes/microbiologia , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Vibrioses/imunologia , Vibrioses/microbiologia , Vibrioses/veterinária , Vibrio alginolyticus/genética , Vibrio parahaemolyticus/genética , Vibrio parahaemolyticus/imunologiaRESUMO
Albizzia julibrissin Durazz, a Chinese Medicine, is commonly used for its anti-anxiety effects. (-)-syringaresnol-4-O-ß-d-apiofuranosyl-(1â2)-ß-d-glucopyranoside (SAG) is the main ingredient of Albizzia julibrissin Durazz. The present study investigated the anxiolytic effect and potential mechanisms on the HPA axis and monoaminergic systems of SAG on acute restraint-stressed rats. The anxiolytic effect of SAG was examined through an open field test and an elevated plus maze test. The concentration of CRF, ACTH, and CORT in plasma was examined by an enzyme-linked immune sorbent assay (ELISA) kit while neurotransmitters in the cerebral cortex and hippocampus of the brain were examined by High Performance Liquid Chromatography (HPLC). We show that repeated treatment with SAG (3.6 mg/kg, p.o.) significantly increased the number and time spent on the central entries in the open-field test when compared to the vehicle/stressed group. In the elevated plus maze test, 3.6 mg/kg SAG could increase the percentage of entries into and time spent on the open arms of the elevated plus maze. In addition, the concentration of CRF, ACTH, and CORT in plasma and neurotransmitters (NE, 5-HT, DA and their metabolites 5-HIAA, DOPAC, and HVA) in the cerebral cortex and hippocampus of the brain were decreased after SAG treatment, as compared to the repeated acute restraint-stressed rats. These results suggest that SAG is a potential anti-anxiety drug candidate.
Assuntos
Albizzia/química , Ansiolíticos/farmacologia , Glicosídeos/farmacologia , Lignanas/farmacologia , Estresse Psicológico/tratamento farmacológico , Ácido 3,4-Di-Hidroxifenilacético/metabolismo , Hormônio Adrenocorticotrópico/sangue , Animais , Monoaminas Biogênicas/metabolismo , Córtex Cerebral/efeitos dos fármacos , Córtex Cerebral/metabolismo , Corticosterona/sangue , Hormônio Liberador da Corticotropina/sangue , Hipocampo/efeitos dos fármacos , Hipocampo/metabolismo , Ácido Homovanílico/metabolismo , Ácido Hidroxi-Indolacético/metabolismo , Masculino , Aprendizagem em Labirinto/efeitos dos fármacos , Atividade Motora/efeitos dos fármacos , Ratos Sprague-Dawley , Estresse Psicológico/metabolismo , Estresse Psicológico/psicologiaRESUMO
In order to study the influence of ecological environment regarding the synthesis and accumulation of metabolites in Eucommiae Cortex, LC-QTOF MS/MS method combined with multivariate statistical analysis was used to analyze the differences of chemical constituents in Eucommiae Cortex from different habitats. Through the analysis of the multistage tandem mass spectrometry, the characteristic peaks were extracted with mass spectrometry data peak matching, peak alignment, and noise filtering. Principal component analysis (PCA) and partial least-squares discriminant analysis (PLS-DA) were used for data processing. The chemical constituents were identified or tentative presumed according to MS accurate mass and MS/MS spectrometry fragmentation information, combined with the software of database search, comparison with reference standards and literature. The results show the differences among samples of Eucommiae Cortex from different habitats are distinguishable. A total of 23 chemical constituents in Eucommiae Cortex were identified or tentative presumed. Among of them, 14 kinds of common differential chemical constituents (aucubin, geniposidic acid, neochlorogenic acid, syringin, olivil-4',4'-di-O-ß-D-glucopyranoside, chlorogenic acid, cryptochlorogenic acid, 1-hydroxypinoresinol- 4',4'-di-O-ß-D-glucopyranoside, caffeic acid, pinoresinol-di-O-ß-D-glucopyranoside, syringaresionl-di-O-ß-D-glucopyranoside, pinoresinol-4'-O-ß-D-glucopyranoside, eucommiol, isochlorogenic acid C and asiatic acid) presented different changing laws. This study provides basic information for revealing the influence law of ecological environment on the biosynthesis of metabolites in Eucommiae Cortex.
Assuntos
Ecossistema , Eucommiaceae/química , Compostos Fitoquímicos/análise , Plantas Medicinais/química , Cromatografia Líquida , Espectrometria de Massas em TandemRESUMO
Natural xanthones have diversity pharmacological activities. Here, a series of xanthones isolated from the pericarps of Garcinia mangostana Linn, named α-Mangostin, 8-Deoxygartanin, Gartanin, Garciniafuran, Garcinone C, Garcinone D, and γ-Mangostin were investigated. Biological screening performed in vitro and in Escherichia coli cells indicated that most of the xanthones exhibited significant inhibition of self-induced ß-amyloid (Aß) aggregation and also ß-site amyloid precursor protein-cleaving enzyme 1, acted as potential antioxidants and biometal chelators. Among these compounds, α-Mangostin, Gartanin, Garcinone C and γ-Mangostin showed better antioxidant properties to scavenge Diphenyl-1-(2,4,6-trinitrophenyl) hydrazyl (DPPH) free radical than Trolox, and potent neuroprotective effects against glutamate-induced HT22 cell death partly by up-regulating HO-1 protein level and then scavenging reactive oxygen species. Moreover, Gartanin, Garcinone C and γ-Mangostin could be able to penetrate the blood-brain barrier (BBB) in vitro. These findings suggest that the natural xanthones have multifunctional activities against Alzheimer's disease (AD) and could be promising compounds for the therapy of AD.
Assuntos
Doença de Alzheimer/metabolismo , Garcinia mangostana , Extratos Vegetais/uso terapêutico , Xantonas/uso terapêutico , Doença de Alzheimer/tratamento farmacológico , Secretases da Proteína Precursora do Amiloide/antagonistas & inibidores , Secretases da Proteína Precursora do Amiloide/metabolismo , Peptídeos beta-Amiloides/antagonistas & inibidores , Peptídeos beta-Amiloides/metabolismo , Animais , Ácido Aspártico Endopeptidases/antagonistas & inibidores , Ácido Aspártico Endopeptidases/metabolismo , Linhagem Celular , Camundongos , Extratos Vegetais/isolamento & purificação , Extratos Vegetais/farmacologia , Xantonas/isolamento & purificação , Xantonas/farmacologiaRESUMO
Oxidative stress mediates the pathogenesis of neurodegenerative disorders. Gartanin, a natural xanthone of mangosteen, possesses multipharmacological activities. Herein, the neuroprotection capacity of gartanin against glutamate-induced damage in HT22 cells and its possible mechanism(s) were investigated for the first time. Glutamate resulted in cell death in a dose-dependent manner and supplementation of 1-10 µM gartanin prevented the detrimental effects of glutamate on cell survival. Additional investigations on the underlying mechanisms suggested that gartanin could effectively reduce glutamate-induced intracellular ROS generation and mitochondrial depolarization. We further found that gartanin induced HO-1 expression independent of nuclear factor erythroid-derived 2-like 2 (Nrf2). Subsequent studies revealed that the inhibitory effects of gartanin on glutamate-induced apoptosis were partially blocked by small interfering RNA-mediated knockdown of HO-1. Finally, the protein expression of phosphorylation of AMP-activated protein kinase (AMPK) and its downstream signal molecules, Sirtuin activator (SIRT1) and peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α), increased after gartanin treatment. Taken together, these findings suggest gartanin is a potential neuroprotective agent against glutamate-induced oxidative injury partially through increasing Nrf-2-independed HO-1 and AMPK/SIRT1/PGC-1α signaling pathways.
Assuntos
Proteínas Quinases Ativadas por AMP/metabolismo , Apoptose/efeitos dos fármacos , Heme Oxigenase-1/metabolismo , Proteínas de Membrana/metabolismo , Fator 2 Relacionado a NF-E2/metabolismo , Neurônios/efeitos dos fármacos , Xantonas/farmacologia , Animais , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Ácido Glutâmico/farmacologia , Camundongos , Neurônios/citologia , Neurônios/metabolismo , Fármacos Neuroprotetores/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais/efeitos dos fármacos , Xantonas/químicaRESUMO
BACKGROUND: Despite several randomized controlled trials comparing operative to nonoperative management of primary patellar dislocation, the optimal management of this condition remains a subject of controversy. The aim of this study was to compare surgical to conservative treatment of outcomes for primary patellar dislocation by meta-analysis all the relative randomized controlled trials. STUDY DESIGN: Meta-analysis. METHODS: After searching multiple online databases (MEDILINE, EMBASE, CLINICAL, OVID, BISOS and Cochrane registry of controlled clinical trials), eight randomized controlled trials including 430 patients were meta-analyzed in which operative treatment was compared with non-operative treatment for primary patellar dislocation. Outcomes evaluated were redislocation rate, Kujala score, episode of instability, Tegner activity score, Hughston visual analog score (VAS) and patient satisfaction. RESULTS: Outcomes on recurrent patellar dislocation (P = 0.004) and Hughston VAS (P = 0.03) were statistically significant in favor of operative management. Tegner activity score (P < 0.00001) was significantly higher in favor of conservative treatment, though only a few studies were identified. There was no significant difference between the two treatments regarding episode of instability (P = 0.41), Kujala score (P = 0.32) or patient satisfaction (P = 0.49). CONCLUSION: Surgical treatment may be better than conservative treatment for patients with primary patellar dislocation on incidence of redislocation. However, since these findings are built on a limited number of studies available, well-designed, multicenter clinical trials with long-term follow-up are required to provide more solid evidence concerning optimal strategies.
Assuntos
Procedimentos Ortopédicos/métodos , Dor , Luxação Patelar/terapia , Satisfação do Paciente , Bases de Dados Factuais , Humanos , Instabilidade Articular/epidemiologia , Complicações Pós-Operatórias/epidemiologia , Ensaios Clínicos Controlados Aleatórios como Assunto , Recidiva , Projetos de PesquisaRESUMO
This work was designed to investigate proteins differentially expressed in cultivated Pseudostellaria heterophylla and its wild type using i TRAQ proteomics approach. The extracted proteins were digested using FASP method and identified by i TRAQ coupled with LC-MS/MS technology and then analyzed by Protein Pilot 5.0 search engine. Proteins differentially expressed were searched through comparison of relatively quantified proteins. The analysis was conducted using GO(gene ontology), KEGG and STRING. A total of 3 775 proteins were detected, among them, 3 676 proteins can be quantified, of which 127 proteins were up-regulated and 205 were down-regulated in cultivated Pseudostellaria heterophylla. We found 71 significantly differentially expressed proteins for further analysis. These proteins were classified into nine categories: heat shock proteins, transferases, oxidoreductases, lyases, isomerases, ligases, hydrolases, tubulin and translocases. The results indicated that the carbohydrate and cellular amino acids metabolism of cultivated Pseudostellaria heterophylla were weaker than its wild type and its ability of responding to stress was much stronger. GWD1, PHS1, GBE1, PGM, and BAM1 are the important proteins to regulate sucrose; met E and CYS are the key proteins that regulate amino acids in cultivated and wild Pseudostellaria heterophylla. This will provide the basic information for exploring the cause of secondary metabolites differences in different ecotype of Pseudostellaria heterophylla and the protein mechanism of its quality formation.