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Isolation and quantification of soil organic matter (SOM) pools under the influence of management practices is needed for assessing the changes in soil fertility. However, the knowledge on how the active, slow and passive pools of SOM respond to long-term fertilization is scarce. Therefore, the present study was designed to isolate the active, slow, and passive pools of soil organic matter through physical fractionation under long-term fertilization. The treatments included; inorganic fertilization (NPK) either alone or combined with a normal dose of manure (MNPK) or a high dose of manure (1.5MNPK) with an unfertilized control (CK) for comparison. The isolated pools were analyzed and compared for their sizes, SOC and TN storage and their contribution to total SOC and TN sequestration. The results revealed that the fertilization enhanced the active, slow and passive pools of SOC and TN and their storage under applied treatments was patterned as 1.5MNK > MNPK > NPK > CK. The highest SOC and TN storage was observed in the active pool, while, greater response to fertilization (in terms of response ratio) was associated with the slow pool. Results show that fertilization enhanced the proportion of SOC and TN stocks to bulk SOC and TN stocks in active and slow pools, while a diminishing trend was found for passive pools. Moreover, the highest response ratio was found for TN sequestration in each pool as compared to SOC, suggesting preferential accumulation of TN over SOC in the studied soil. Nevertheless, the highest SOC and TN storage took place in the active pool. The slow pool showed greater response to applied fertilizer, with the highest values being observed under 1.5MNPK. This study concluded that long-term manure + inorganic fertilization is crucial for enhancing C and N sequestration by altering the size and response of SOM pools.
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Nitrogênio , Solo , Nitrogênio/análise , Carbono/análise , Esterco , Agricultura/métodos , Fertilizantes/análise , Fertilização , ChinaRESUMO
Long-term fertilization has shown a high relevance as regards soil organic carbon (SOC) sequestration, but the degree of stability of the sequestered SOC has not been widely studied up to now. Using physical fractionation combined with laboratory incubation and NMR spectroscopy, we evaluated the differences in SOC stability caused by long-term fertilization. Four SOC fractions were isolated and examined for contents and chemical composition and cumulative amount of CO2-C respired from the fractions under six fertilization treatments: control (CK); balanced inorganic fertilization (NPK); NPK combined with pig manure (MNPK); NPK combined 1.5 times of pig manure (1.5MNPK); and NPK combined with high amount of manure (M2NPK). The highest contents of SOC were recorded for the coarse particulate organic carbon (cPOC) fraction, ranging from 17.25 to 30.47 g kg-1 under CK and M2NPK. The highest cumulative amount of CO2-C was released from the cPOC fraction under manure treatments (M2NPK and 1.5NPKM), which was 56 and 43% higher than that from CK, whereas the lowest amount of CO2-C was released from the mineral associated-C (MOC) fraction under the same treatments, being 65 and 49% higher than that released from CK, suggesting low SOC stability in cPOC and high SOC stability in MOC fractions. However, manure treatments (M2NPK and 1.5NPKM) greatly lowered the specific amount of C-mineralized (C-mineralized per unit total SOC) in fractions and whole soil, suggesting the ability of manure to accumulate more SOC by reducing SOC losses. Moreover, carbonyl-C was found to be the form of SOC experiencing major degree of sequestration under current fertilization practices. The SOC stability indices; aromaticity index (AI), hydrophobicity index (HI) and alkyl-C/O-alkyl-C were found to be higher in manure treated plots further suggesting higher stability of SOC under manure addition. Thus, long-term manure combined with mineral fertilizers would enhance SOC stability through minimizing SOC losses and promoting accumulation of stable C forms in a Chinese Mollisol.
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Carbono , Solo , Agricultura/métodos , Animais , Carbono/análise , Fertilização , Fertilizantes/análise , Espectroscopia de Ressonância Magnética , Esterco/análise , Solo/química , SuínosRESUMO
The CRISPR/Cas9 site-directed gene-editing system offers great advantages for identifying gene function and crop improvement. The circadian clock measures and conveys day length information to control rhythmic hypocotyl growth in photoperiodic conditions, to achieve optimal fitness, but operates through largely unknown mechanisms. Here, we generated core circadian clock evening components, Brassica rapa PSEUDO-RESPONSE REGULATOR (BrPRR) 1a, 1b, and 1ab (both 1a and 1b double knockout) mutants, using CRISPR/Cas9 genome editing in Chinese cabbage, where 9-16 genetic edited lines of each mutant were obtained. The targeted deep sequencing showed that each mutant had 2-4 different mutation types at the target sites in the BrPRR1a and BrPRR1b genes. To identify the functions of BrPRR1a and 1b genes, hypocotyl length, and mRNA and protein levels of core circadian clock morning components, BrCCA1 (CIRCADIAN CLOCK-ASSOCIATED 1) and BrLHY (LATE ELONGATED HYPOCOTYL) a and b were examined under light/dark cycles and continuous light conditions. The BrPRR1a and 1ab double mutants showed longer hypocotyls, lower core circadian clock morning component mRNA and protein levels, and a shorter circadian rhythm than wildtype (WT). On the other hand, the BrPRR1b mutant was not significantly different from WT. These results suggested that two paralogous genes may not be associated with the same regulatory function in Chinese cabbage. Taken together, our results demonstrated that CRISPR/Cas9 is an efficient tool for achieving targeted genome modifications and elucidating the biological functions of circadian clock genes in B. rapa, for both breeding and improvement.
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Brassica rapa , Brassica , Brassica/genética , Brassica rapa/genética , Sistemas CRISPR-Cas , China , Ritmo Circadiano/fisiologia , Regulação da Expressão Gênica de Plantas , Mutagênese , Melhoramento Vegetal , RNA MensageiroRESUMO
Soil aggregation plays a critical role in the maintenance of soil structure, as well as in its productivity. Fertilization influences soil aggregation, especially by regulating soil organic carbon (SOC) and total nitrogen (TN) contents in aggregate fractions. The present study evaluated the influence of three contrasting fertilizer regimes (unfertilized control -CK-, mineral fertilization -NPK- and manure combined with NPK -NPKM) on soil aggregate stability, aggregate-associated organic carbon and total nitrogen sequestration and mineralization of SOC. Soil samples from (20 cm) depth were collected from a long-term fertilization experiment and analysed for size distribution ranging (>250 µm, 250-53 µm and <53 µm sizes), SOC and TN contents, as well as for mineralization of bulk and aggregate associated-SOC. Both NPK and NPKM fertilizations significantly enhanced SOC and TN contents in bulk soil and its constituent aggregates of >250 µm, 250-53 µm and <53 µm sizes, as compared to CK. Long-term NPK and NPKM increased SOC and TN stock in bulk soil by 45 and 98%, and by 70 and 144%, respectively, as compared to CK. Similarly, higher values of SOC and TN stock in all aggregate fractions was observed with the application of NPKM. Application of NPK and NPKM for 26 years significantly increased aggregate stability, which was positively correlated with total SOC contents in terms of mean weight diameter (MWD) (Adj. R2 = 0.689, p < 0.03) and geometric mean diameter (GMD) (Adj. R2 = 0.471, p < 0.24). Moreover, higher scores regarding cumulative mineralization for bulk soil and aggregate associated OC were observed with the application of NPK and NPKM. Irrespective of treatments, higher cumulative C-mineralization was observed for macro-aggregates (>250 µm size) followed by 250-53 µm and <53 µm size aggregates. Interestingly, a highly positive correlation was observed between aggregate stability and the cumulative amount of mineralization for bulk soil and aggregate fractions, with R2 ranging from 0.84 to 0.99. This study evidenced that long-term fertilization of NPK and NPKM can improve soil aggregation, stability and associated OC and TN stock in aggregates, as well as aggregate-associated OC mineralization, which was further governed by aggregate size.
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Nitrogênio/análise , Solo , Agricultura , Carbono/análise , China , Fertilizantes/análise , EstercoRESUMO
Autoantigen-specific immunotherapy promises effective treatment for devastating tissue specific autoimmune diseases like multiple sclerosis (MS) and type 1 diabetes (T1D). Because activated dendritic cells (DCs) stimulate the differentiation of autoreactive T cells involved in the initiation of autoimmunity, blocking the activation of DCs may be an effective strategy for inhibiting tissue specific autoimmunity. Following this approach, immature DCs were shown to remain inactive after treatment with chimeric fusion proteins composed of the cholera toxin B subunit adjuvant linked to autoantigens like proinsulin (CTB-INS). Mass spectrometer analysis of human DCs treated with CTB-INS suggest that upregulation of the tryptophan catabolic enzyme indoleamine 2, 3-dioxygenase (IDO1) is responsible for inhibiting DC activation thereby resulting in a state of immunological tolerance within the DC. Here we show that the fusion protein CTB-INS inhibits human monocyte derived DC (moDC) activation through stimulation of IDO1 biosynthesis and that the resultant state of DC tolerance can be further enhanced by the presence of residual E. coli lipopolysaccharide (LPS) present in partially purified CTB-INS preparations. Additional experiments showed that LPS enhancement of DC tolerance was dependent upon stimulation of IDO1 biosynthesis. LPS stimulation of increased levels of IDO1 in the DC resulted in increased secretion of kynurenines, tryptophan degradation products known to suppress DC mediated pro-inflammatory T cell differentiation and to stimulate the proliferation of regulatory T cells (Tregs). Further, the presence of LPS in CTB-INS treated DCs stimulated the biosynthesis of costimulatory factors CD80 and CD86 but failed to upregulate maturation factor CD83, suggesting CTB-INS treated DCs may be maintained in a state of semi-activation. While treatment of moDCs with increasing amounts of LPS free CTB-INS was shown to increase DC secretion of the anti-inflammatory cytokine IL-10, the presence of residual LPS in partially purified CTB-INS preparations dramatically increased IL-10 secretion, suggesting that CTB-INS may enhance DC mediated immunological tolerance by stimulating the proliferation of anti-inflammatory T cells. While the extraction of LPS from bacterial generated CTB-INS may remove additional unknown factors that may contribute to the regulation of IDO1 levels, together, our experimental data suggest that LPS stimulates the ability of CTB-INS to induce IDO1 and IL-10 important factors required for establishment of a state of functional immunological tolerance in human DCs. Regulation of the ratio of LPS to CTB-INS may prove to be an effective method for optimization of readily available "off the shelf" CTB-INS mediated immune-therapy for tissue specific autoimmune diseases including type 1 diabetes.
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Autoantígenos/metabolismo , Doenças Autoimunes/terapia , Toxina da Cólera/metabolismo , Células Dendríticas/imunologia , Imunoterapia/métodos , Proinsulina/metabolismo , Proteínas Recombinantes de Fusão/metabolismo , Doenças Autoimunes/imunologia , Diferenciação Celular , Células Cultivadas , Toxina da Cólera/genética , Humanos , Tolerância Imunológica , Indolamina-Pirrol 2,3,-Dioxigenase/genética , Indolamina-Pirrol 2,3,-Dioxigenase/metabolismo , Interleucina-10/genética , Interleucina-10/metabolismo , Lipopolissacarídeos/metabolismo , Proinsulina/genética , RNA Interferente Pequeno/genética , Proteínas Recombinantes de Fusão/genéticaRESUMO
Cyclic citrullinated peptide (CCP) antibody has been shown recently to be a promising marker for early detection and diagnosis of rheumatoid arthritis (RA). In order to exploit newly developed therapies for RA, early intervention is crucial in preventing irreversible joint damage. Here, we describe use of a plant expression system to produce a CCP antibody that could be used in the early diagnosis of RA. Heavy and light chain gene sequences of a CCP monoclonal antibody (CCP mAb) were cloned from the hybridoma cell (12G1) and introduced into two separate plant expression vectors under the control of the rice α-amylase 3D (RAmy3D) promoter system. The vectors were introduced into rice calli (Oryza sativa L. cv. Dongjin) using Agrobacterium tumefaciens mediated transformation. Integration of the CCP mAb genes into rice chromosomes was confirmed by a genomic DNA polymerase chain reaction and expression was verified by northern blot analysis of mRNA. The in vivo assembly and secretion of CCP mAb occurred in transgenic rice cell suspension culture under the RAmy3D expression system; accumulated CCP mAbs in the medium were purified by protein G affinity chromatography. Immunoblot assays and ELISA showed these plant-produced CCP mAbs successfully bound to a synthetic CCP antigen. Taken together, our results suggest that CCP mAb produced in a transgenic rice suspension culture were easily purified and biologically active against their antigen in the RA, and thus may be used a specific serological marker, which is present very early in the RA.
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Anticorpos Monoclonais/metabolismo , Oryza/imunologia , Peptídeos Cíclicos/imunologia , Plantas Geneticamente Modificadas/imunologia , Agrobacterium tumefaciens/genética , Anticorpos Monoclonais/genética , Anticorpos Monoclonais/imunologia , Técnicas de Cultura de Células , Células Cultivadas , Vetores Genéticos , Humanos , Oryza/genética , Oryza/metabolismo , Peptídeos Cíclicos/genética , Peptídeos Cíclicos/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismoRESUMO
Gaucher disease is an inherited metabolic disease caused by genetic acid ß -glucosidase (GBA) deficiency and is currently treated by enzyme replacement therapy. For uptake into macrophages, GBA needs to carry terminal mannose residues on their N-glycans. Knockout mutant rice of N-acetylglucosaminyltransferase-I (gnt1) have a disrupted N-glycan processing pathway and produce only glycoproteins with high mannose residues. In this study, we introduced a gene encoding recombinant human GBA into both wild-type rice (WT) and rice gnt1 calli. Target gene integration and mRNA expression were confirmed by genomic DNA PCR and Northern blotting, respectively. Secreted rhGBAs in culture media from cell lines originating from both WT (WT-GBA) and rice gnt1 (gnt1-GBA) were detected by Western blotting. Each rhGBA was purified by affinity and ion exchange chromatography. In vitro catalytic activity of purified rhGBA was comparable to commercial Chinese hamster ovary cell-derived rhGBA. N-glycans were isolated from WT-GBA and gnt1-GBA and analyzed by using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The amounts of high mannose-type N-glycans were highly elevated in gnt1-GBA (100%) compared to WT-GBA (1%).
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Doença de Gaucher/tratamento farmacológico , Glucosilceramidase , Mutação , Oryza , Plantas Geneticamente Modificadas , Polissacarídeos , Animais , Células CHO , Cricetulus , Glucosilceramidase/biossíntese , Glucosilceramidase/genética , Glucosilceramidase/isolamento & purificação , Glucosilceramidase/uso terapêutico , Humanos , Oryza/química , Oryza/genética , Oryza/metabolismo , Plantas Geneticamente Modificadas/química , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Polissacarídeos/química , Polissacarídeos/genética , Polissacarídeos/metabolismo , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificaçãoRESUMO
High levels of serum long chain saturated fatty acids (LCSFAs) have been associated with inflammation in type 2 diabetes. Dietary SFAs can promote inflammation, the secretion of IgG antibodies, and secretion of the proinflammatory cytokine IL-1ß. This study characterizes anti-LCSFA IgG antibodies from patients with type 2 diabetes. Serum samples from several cohorts with type 2 diabetes were analyzed for the presence of anti-LCSFA IgG, the cytokine IL-1ß, and nonesterified fatty acids. Anti-LCSFA IgG was isolated from patient samples and used for in vitro characterization of avidity and specificity. A cohort participating in En Balance, a diabetes health education program that improved diabetes management, tested positive for anti-LCSFA IgG. Following the 3-month program, the cohort showed a significant reduction in anti-LCSFA IgG levels. Anti-LCSFA antibodies isolated from these patients demonstrated high avidity, were specific for long chain SFAs, and correlated with serum fatty acids in patients with managed type 2 diabetes. Interestingly, anti-LCSFA IgG neutralized PA-induced IL-1ß secretion by dendritic cells. Our data shows that nonesterified SFAs are recognized by IgG antibodies present in human blood. The identification of anti-LCSFA IgG antibodies in human sera establishes a basis for further exploration of lipid induced immune responses in diabetic patients.
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Diabetes Mellitus Tipo 2/imunologia , Ácidos Graxos/imunologia , Imunoglobulina G/sangue , Adulto , Idoso , Especificidade de Anticorpos , Células Dendríticas/imunologia , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Interleucina-1beta/sangue , Masculino , Pessoa de Meia-Idade , Ácido Palmítico/imunologiaRESUMO
We focused on the geminiviral vector systems to develop an efficient vector system for plant biotechnology. Begomoviruses and curtoviruses, which belong to the Geminiviridae family, contain an intergenic region (IR) and four genes involved in replication, including replication-associated protein (Rep, C1), transcriptional activator (TrAP, C2), and replication enhancer (REn, C3). Geminiviruses can amplify thousands of copies of viral DNA using plant DNA polymerase and viral replication-related enzymes and accumulate viral proteins at high concentrations. In this study, we optimized geminiviral DNA replicon vectors based on tomato yellow leaf curl virus (TYLCV), honeysuckle yellow vein virus (HYVV), and mild curly top virus (BMCTV) for the rapid, high-yield plant-based production of recombinant proteins. Confirmation of the optimal combination by co-delivery of each replication-related gene and each IR harboring the Pontellina plumata-derived turbo green fluorescence protein (tGFP) gene via agroinfiltration in Nicotiana benthamiana leaves resulted in efficient replicon amplification and robust protein production within 3 days. Co-expression with the p19 protein of the tomato bush stunt virus, a gene-silencing suppressor, further enhanced tGFP accumulation by stabilizing mRNA. With this system, tGFP protein was produced at 0.7-1.2 mg/g leaf fresh weight, corresponding to 6.9-12.1% in total soluble protein. These results demonstrate the advantages of rapid and high-level production of recombinant proteins using the geminiviral DNA replicon system for transient expression in plants.
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Major depressive disorder (MDD) is one of the foremost causes of disability and premature death worldwide. Although the available antidepressants are effective and well tolerated, they also have many limitations. Therapeutic advances in developing a new drug's ultimate relation between MDD and chronobiology, which targets the circadian rhythm, led to a renewed focus on psychiatric disorders. In order to provide a critical analysis about antidepressant properties of agomelatine, a detailed PubMed (Medline), Scopus (Embase), Web of Science (Web of Knowledge), Cochrane Library, Google Scholar, and PsycInfo search was performed using the following keywords: melatonin analog, agomelatine, safety, efficacy, adverse effects, pharmacokinetics, pharmacodynamics, circadian rhythm, sleep disorders, neuroplasticity, MDD, bipolar disorder, anhedonia, anxiety, generalized anxiety disorder (GAD), and mood disorders. Agomelatine is a unique melatonin analog with antidepressant properties and a large therapeutic index that improves clinical safety. Published articles revealed that agomelatine is a melatonin receptors (MT1 and MT2) agonist and 5HT2C receptor antagonist. The effects receptors' on melatonin receptors enable the resynchronization of irregular circadian rhythms with beneficial effects on sleep architectures. In this way, agomelatine is accredited for its unique mode of action, which helps to exert antidepressant effects and resynchronize the sleep-wake cycle. To sum up, an agomelatine has not only antidepressant properties but also has anxiolytic effects.
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Transtorno Depressivo Maior , Melatonina , Acetamidas , Antidepressivos/efeitos adversos , Transtorno Depressivo Maior/induzido quimicamente , Transtorno Depressivo Maior/tratamento farmacológico , Humanos , Melatonina/farmacologia , Melatonina/uso terapêutico , Naftalenos , Receptores de Melatonina/uso terapêuticoRESUMO
Glucosinolates (GSLs) are secondary metabolites providing defense against pathogens and herbivores in plants, and anti-carcinogenic activity against human cancer cells. Profiles of GSLs vary greatly among members of genus Brassica. In this study, we found that a reference line of Chinese cabbage (B. rapa ssp. pekinensis), 'Chiifu' contains significantly lower amounts of total GSLs than the oilseed-type B. rapa (B. rapa ssp. trilocularis) line 'LP08'. This study aimed to identify the key regulators of the high accumulation of GSLs in Brassica rapa plants using transcriptomic and linkage mapping approaches. Comparative transcriptome analysis showed that, in total, 8,276 and 9,878 genes were differentially expressed between 'Chiifu' and 'LP08' under light and dark conditions, respectively. Among 162 B. rapa GSL pathway genes, 79 were related to GSL metabolism under light conditions. We also performed QTL analysis using a single nucleotide polymorphism-based linkage map constructed using 151 F5 individuals derived from a cross between the 'Chiifu' and 'LP08' inbred lines. Two major QTL peaks were successfully identified on chromosome 3 using high-performance liquid chromatography to obtain GSL profiles from 97 F5 recombinant inbred lines. The MYB-domain transcription factor gene BrMYB28.1 (Bra012961) was found in the highest QTL peak region. The second highest peak was located near the 2-oxoacid-dependent dioxygenase gene BrGSL-OH.1 (Bra022920). This study identified major genes responsible for differing profiles of GSLs between 'Chiifu' and 'LP08'. Thus, our study provides molecular insights into differences in GSL profiles between vegetative- and oilseed-type B. rapa plants.
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A synthetic bovine trypsinogen (sbTrypsinogen) was synthesized on the basis of rice-optimized codon usage via an overlap PCR strategy, prior to being expressed under the control of the sucrose starvation-inducible rice α-amylase 3D (RAmy3D) promoter. Secretion of trypsin into the culture medium was achieved by using the existing signal peptide. The plant expression vector was introduced into rice calli (Oryza sativa L. cv. Dongjin), mediated by Agrobacterium tumefaciens. The integration of the sbTrypsinogen gene into the chromosome of the transgenic rice callus was verified via genomic DNA PCR amplification, and sbTrypsin expression in transgenic rice suspension cells was confirmed via Northern blot analysis. Western blot analysis detected glycosylated proteins in the culture medium, having masses from 24 to 26 kDa, following induction by sugar starvation. Proteolytic activity of the rice-derived trypsin was confirmed by gelatin zymogram, and was similar to that of the commercial bovine-produced trypsin. The yields of sbTrypsin that accumulated in the transgenic rice cell suspension medium were 15 mg/L at 5 days after sugar starvation.
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Oryza , Plantas Geneticamente Modificadas , Proteínas Recombinantes/biossíntese , Tripsina/biossíntese , Sequência de Aminoácidos , Animais , Sequência de Bases , Bovinos , Células Cultivadas , Meios de Cultivo Condicionados , Vetores Genéticos , Dados de Sequência Molecular , Tripsinogênio/biossínteseRESUMO
Crop productivity and soil health are limited by organic carbon (OC), however, the variations in the mechanisms of SOC preservation in a complete soil profile subjected to long-term fertilization remains unclear. The objective of the study was to examined the content and profile distribution of the distinctive SOC protection mechanisms on a complete profile (0-100 cm) of Eumorthic Anthrosols in Northwest China after 23 years of chemical and manure fertilization. The soil was fractionated by combined physical-chemical and density floatation techniques. Throughout the profile, significant variations were observed among fractions. In the topsoil (0-20 and 20-40 cm), mineral coupling with the fertilization of manure (MNPK) enhanced total SOC content and recorded for 29% of SOC in the 0-20 and 20-40 cm layers. Moreover, MNPK increased the SOC content of the unprotected cPOC fraction by 60.9% and 61.5% in the 0-20 and 20-40 cm layer, while SOC content was low in the subsoil layers (40-60, 60-80 and 80-100 cm, respectively) compared with the control (C). The highest OC under MNPK in physically protected micro-aggregates (µagg) (6.36 and 6.06 g C kg-1), and occluded particulate organic carbon (iPOC) (1.41 and 1.29 g C kg-1) was found in the topsoil layers. The unprotected cPOC fraction was the greatest C accumulating fraction in the topsoil layers, followed by µagg and H-µSilt fractions in the soil profile, implying that these fractions were the most sensitive to the fertilization treatments. Overall, the unprotected, physically protected, and physico-chemically protected fractions were the dominant fractions for the sequestration of carbon across fertilization treatments and soil layers.
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Carbono , Solo , Agricultura , Povo Asiático , Carbono/análise , China , Fertilização , Fertilizantes , HumanosRESUMO
Circadian clocks integrate environmental cues with endogenous signals to coordinate physiological outputs. Clock genes in plants are involved in many physiological and developmental processes, such as photosynthesis, stomata opening, stem elongation, light signaling, and floral induction. Many Brassicaceae family plants, including Chinese cabbage (Brassica rapa ssp. pekinensis), produce a unique glucosinolate (GSL) secondary metabolite, which enhances plant protection, facilitates the design of functional foods, and has potential medical applications (e.g., as antidiabetic and anticancer agents). The levels of GSLs change diurnally, suggesting a connection to the circadian clock system. We investigated whether circadian clock genes affect the biosynthesis of GSLs in Brassica rapa using RNAi-mediated suppressed transgenic Brassica rapa GIGENTEA homolog (BrGI knockdown; hereafter GK1) Chinese cabbage. GIGANTEA plays an important role in the plant circadian clock system and is related to various developmental and metabolic processes. Using a validated GK1 transgenic line, we performed RNA sequencing and high-performance liquid chromatography analyses. The transcript levels of many GSL pathway genes were significantly altered in GK1 transgenic plants. In addition, GSL contents were substantially reduced in GK1 transgenic plants. We report that the BrGI circadian clock gene is required for the biosynthesis of GSLs in Chinese cabbage plants.
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Brassica rapa/genética , Proteínas CLOCK/genética , Glucosinolatos/metabolismo , Brassica rapa/metabolismo , China , Relógios Circadianos/genética , Alimento Funcional , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Glucosinolatos/genética , Redes e Vias Metabólicas/genética , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas , Transcriptoma/genéticaRESUMO
OBJECTIVE: To investigate the effects of different methods of treatment, including tonifying the kidney, activating blood and soothing the liver, on mice with autoimmune premature ovarian failure (POF). METHODS: After preparing ovarian antigen with the ovarian tissue of female BALB/C mice, POF in female BALB/C mice was induced by immunization injection of the ovarian antigen on multiple subcutaneous sites and two posterior soles. The mice were treated with Liuwei Dihuang Pill, Shaofu Zhuyu Granule, Chaihu Shugan Pill respectively after the third immunization. Levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E(2)), prolactin (PRL), testosterone (T), anti-ovarian antibodies (AOAb) and anti-zona pellucida antibody (aZP) in sera were analyzed by enzyme-linked immunosorbent assay. RESULTS: Compared with the normal group, the levels of serum FSH and LH were increased and the level of E(2) was decreased in the untreated group. AOAb and aZP positive rates in the untreated group were obviously higher than those in the normal group. Liuwei Dihuang Pill could significantly reduce the levels of FSH and LH and increase the level of E(2) as compared with the untreated group, but there were no significant differences in the AOAb and aZP positive rates. Shaofu Zhuyu Granule and Chaihu Shugan Pill could significantly reduce the positive rates of AOAb and aZP as compared with the untreated group, but no significant effect on the levels of FSH, LH and E(2) were observed. The drugs had no effects on serum contents of PRL and T. CONCLUSION: Tonifying the kidney method can recover the ovarian function in POF mice mainly by regulating the hormone levels. Activating blood and soothing the liver methods can improve the humoral immune function in POF mice.
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Doenças Autoimunes/tratamento farmacológico , Medicamentos de Ervas Chinesas/uso terapêutico , Fitoterapia , Extratos Vegetais/uso terapêutico , Insuficiência Ovariana Primária/tratamento farmacológico , Animais , Feminino , Medicina Tradicional Chinesa , Camundongos , Camundongos Endogâmicos BALB C , Insuficiência Ovariana Primária/etiologiaRESUMO
Heart failure combined with pulmonary infection is characterized by heart-lung qi deficiency, spleen-kidney yang deficiency, and deficiency of both qi and yin as the the root of disease; lung heat, phlegm heat, and blood stasis as the manifestation of disease. Thus, clearing, dispersing, supplementing, and transforming as the main treatment methods were proposed. “Clearing” refers to the method of clearing heat to release the exterior, clearing heat from the ying level to relieve heat, and clearing heat and cooling blood, based on the degree of differentiation of pathogenic heat and stagnation of qi and blood; “dispersing” mainly refers to the method of diffusing the lung and dredging the bowels; “supplementing” refers to the method of relieving fever with sweet-warm medicinals, nourishing yin and tonifying qi, and warming yang and transforming qi; “transforming” refers to the method of resolving phlegm and dampness, invigorating blood and dissolving stasis according to different pathological factors.
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The term ''panvascular'' refers to the human vascular system, which is a complex network of arteries, veins, and lymphatic vessels. Panvascular diseases refer to a group of vascular system diseases, with vascular atherosclerosis as the common pathological feature. The panvascular diseases in target organs such as the heart, brain, kidney, and limbs are caused by ischemia or bleeding, including arterial system diseases, venous system diseases, microcirculation system diseases, and Zangfu organ-blood vessel diseases. The concept of panvascular diseases integrates vascular lesions and target organ damage. In clinical practice, blood vessels in multiple regions are regarded as a large vascular unit system, and vascular lesions and the induced target organ damage are considered as a whole. Based on the holistic concept and the Zangxiang theory in traditional Chinese medicine (TCM), the ''blood vessel-Zangfu organ-syndrome differentiation and treatment'' network is built, on the basis of which a pattern of vascular disease-Zangfu organ dysfunction-syndrome differentiation and treatment is applied to the TCM diagnosis and treatment of panvascular diseases. The theory of treating arterial system diseases from the heart, venous system diseases from the kidneys, and microvascular system diseases from the liver is proposed. According to the causes identified based on syndrome differentiation, this paper summarizes the methods of reinforcing Yang and activating blood (including warming Yang and activating blood, replenishing Qi and activating blood, replenishing Qi, nourishing Yin and activating blood, activating Yang and blood, dispersing cold and activating blood), cooling blood and resolving stasis, tonifying kidney and promoting urination coupled with activating blood and dredging vessels, nourishing Yin and tonifying kidney coupled with activating blood and dredging vessels, and soothing liver and regulating Qi coupled with activating blood and dredging collaterals, as well as wind-extinguishing medicines, applied to the treatment of panvascular diseases, aiming to provide methods and ideas for the treatment of vascular diseases with TCM.
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AIM:To rapidly evaluate the effective-ness,safety and economy of dienogest in the treat-ment of endometriosis so as to provide evidence-based reference for clinical drug selection and deci-sion.METHODS:Retrieved from PubMed,Embase,Cochrane Library,CNKI,SinoMed,Wanfang and health technology assessment(HTA)organization websites,HTA report,systematic review/Meta-anal-ysis and pharmacoeconomic study were included during the inception to Sep 2023.Data extraction and quality evaluation were carried out for the lit-erature that met the inclusion and exclusion crite-ria,and the research results were summarized and analyzed qualitatively.RESULTS:Nine systematic re-view/Meta-analyses,and 2 pharmacoeconomic studies were included.The quality of Meta-analysis literatures was low,and the quality of economic re-search was good.In terms of effectiveness,com-pared with no treatment,dienogest can significant-ly reduce postoperative recurrence rate,VAS score,and improve pregnancy rate and effective rate;Di-enogest has similar therapeutic effects in terms of recurrence rate as GnRHa and COC;With regard to recurrence rate and pregnancy rate,dienogest is superior to danazol,gestrinone,and mifepristone.In terms of safety,compared with placebo,the inci-dence of vaginal bleeding and headache was signifi-cantly increased with dienogest,and there was no difference in bone loss;The incidence of vaginal bleeding caused by dienogest was significantly high-er than that of GnRHa,but the incidence of hot flashes and bone loss was lower.In terms of econo-my,dienogest has a more cost-effective advantage comparing with GnRHa,but does not have econom-ic benefits comparing with COC.CONCLUSION:Di-enogest has good effectiveness and safety in the treatment of endometriosis,and economically su-perior to GnRHa,but inferior to COC.
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Endometriosis is a chronic,relapsing disease that requires long-term management.Ado-lescent endometriosis is a group that can be ig-nored,and the drug treatment is an important method for long-term management and fertility preservation.There are many kinds of drugs to treat endometriosis,such as NSAIDs,combined oral contraceptives(COC),progesterone,GnRHa.For the drug treatment selection,the characteris-tics of adolescents should be carefully considered and individualized treatment should be provided.In order to better long-term management,it is nec-essary to develop new treatments,which can effec-tively relieve endometriosis without damaging fer-tility.This review focuses on the drug treatment op-tions and research progress of basic drug treat-ment for adolescent endometriosis,so as to delay disease progression,reduce adverse reactions,pre-serve fertility,and improve quality of life.
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Objective To evaluate the clinical practical value of the golden angle radial sparse parallel(GRASP)technology in the MR imaging quality of lung cancer under free-breathing state.Methods The imaging data of 30 lung cancer patients diagnosed with CT screening were collected by using a large-aperture 3.0T MR scanner.All patients underwent routine Cartesian breath-hold volume interpolation(BH-VIBE)scanning,then free-breathing dynamic contrast-enhanced magnetic resonance imaging(DCE-MRI)scanning based on GRASP technology,and another repetitive BH-VIBE scanning.The MR imaging quality was independently evaluated by two experienced thoracic radiologists including overall image quality,lesion outline,artifact grade and diagnostic confidence,mean-while,the CT images of the patient at the same time were used as a reference.The sequence information on MR images was also pro-cessed blindly.Results The GRASP sequence could effectively eliminate the interference of motion artifacts such as respiration and cardiac large vessel beats,and the image quality,lesion outline,artifact grade,and diagnostic confidence of the GRASP sequence were significantly better than the BH-VIBE(P<0.01).The image quality of the GRASP sequence was more satisfactory with clea-rer tumor boundary and more details.Conclusion Free-breathing high-temporal-spatial resolution GRASP technology has better image quality than traditional BH-VIBE scanning,and it is a reliable method for high-resolution imaging of lung cancer.