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Abstract Light intensity is a key factor affecting the synthesis of secondary metabolites in plants. However, the response mechanisms of metabolites and genes in Gentiana macrophylla under different light intensities have not been determined. In the present study, G. macrophylla seedlings were treated with LED light intensities of 15 µmol/m2/s (low light, LL), 90 µmol/m2/s (medium light, ML), and 200 µmol/m2/s (high light, HL), and leaves were collected on the 5th day for further investigation. A total of 2162 metabolites were detected, in which, the most abundant metabolites were identified as flavonoids, carbohydrates, terpenoids and amino acids. A total of 3313 and 613 differentially expressed genes (DEGs) were identified in the LL and HL groups compared with the ML group, respectively, mainly enriched in KEGG pathways such as carotenoid biosynthesis, carbon metabolism, glycolysis/gluconeogenesis, amino acids biosynthesis, plant MAPK pathway and plant hormone signaling. Besides, the transcription factors of GmMYB5 and GmbHLH20 were determined to be significantly correlated with loganic acid biosynthesis; the expression of photosystem-related enzyme genes was altered under different light intensities, regulating the expression of enzyme genes involved in the carotenoid, chlorophyll, glycolysis and amino acids pathway, then affecting their metabolic biosynthesis. As a result, low light inhibited photosynthesis, delayed glycolysis, thus, increased certain amino acids and decreased loganic acid production, while high light got an opposite trend. Our research contributed significantly to understand the molecular mechanism of light intensity in controlling metabolic accumulation in G. macrophylla.

Abstract Gentiana macrophylla is one of Chinese herbal medicines in which 4 kinds of iridoids or secoiridoids, such as loganic acid, sweroside, swertiamarin, and gentiopicroside, are identified as the dominant medicinal secondary metabolites. WRKY, as a large family of transcription factors (TFs), plays an important role in the synthesis of secondary metabolites in plants. Therefore, WRKY genes involved in the biosynthesis of secoiridoids in G. macrophylla were systematically studied. First, a comprehensive genome-wide analysis was performed, and 42 GmWRKY genes were identified, which were unevenly distributed in 12 chromosomes. Accordingly, gene structure, collinearity, sequence alignment, phylogenetic, conserved motif and promoter analyses were performed, and the GmWRKY proteins were divided into three subfamilies based on phylogenetic and multiple sequence alignment analyses. Moreover, the enzyme-encoding genes of the secoiridoid biosynthesis pathway and their promoters were then analysed, and the contents of the four secoiridoids were determined in different tissues. Accordingly, correlation analysis was performed using Pearson′s correlation coefficient to construct WRKY gene-enzyme-encoding genes and WRKY gene–metabolite networks. Meanwhile, G. macrophylla seedlings were treated with methyl jasmonate (MeJA) to detect the dynamic change trend of GmWRKYs, biosynthetic genes, and medicinal ingredient accumulation. Thus, a total of 12 GmWRKYs were identified to be involved in the biosynthesis of secoiridoids, of which 8 (GmWRKY1, 6, 12, 17, 33, 34, 38 and 39) were found to regulate the synthesis of gentiopicroside, and 4 (GmWRKY7, 14, 26 and 41) were found to regulate the synthesis of loganic acid. Taken together, this study systematically identified WRKY transcription factors related to the biosynthesis of secoiridoids in G. macrophylla, which could be used as a cue for further investigation of WRKY gene functions in secondary metabolite accumulation.

Soil carbon mineralization plays an important role in the carbon cycle of terrestrial ecosystems. When it comes to the soil carbon cycle, however, research on how carbon mineralization characteristics of fertilized Camellia oleifera forest soil respond to temperature changes remains limited. This study used an indoor constant temperature incubation method to examine the effects of the vermicomposting of cow dung by applying it at three different quantities (A: 0.8 kg earthworm + 62.5 kg cow dung/Camellia oleifera; B: 1.6 kg earthworm + 125 kg cow dung/Camellia oleifera; C: 2.4 kg earthworm + 187.5 kg cow dung/Camellia oleifera) and set a control group with Camellia oleifera forest not being fertilized (CK). This research was conducted with incubators set at 5 °C, 15 °C, 25 °C, and 35 °C, and with continuous monitoring of soil carbon mineralization characteristics and temperature sensitivity of organic carbon mineralization. The results showed significant increases in soil MBC, MBN, DOC, DON, NO3−-N, and NH4+-N in groups with applications of cow-dung vermicomposting compared to CK. Except at 35 °C, soil respiration in the Camellia oleifera of Group A was consistently the strongest. The maximum soil carbon emission (C0) was determined through a simulation of potential carbon emissions, with all correlation coefficients exceeding 0.95. The contents of TC and TN were positively correlated with MBC and MBN (p micro was negatively correlated with TN, AN, MBN, and inorganic nitrogen. Based on temperature sensitivity (Q10), the influence of temperature on soil mineralization rate was observed. The vermicomposting of cow dung had a noticeable effect, as Group B showed significantly stronger enzyme activity compared to other groups. These results indicate that changes in MBC can impact the stability of soil carbon mineralization. The roles of soil moisture and microorganisms should be considered when predicting dynamic changes in the soil carbon pool of Camellia oleifera when applying fertilizers and improving its soil carbon sequestration capacity.

By examining the evolution of pre-Qin Confucian thought on the Heaven–human relationship, and a new interpretive perspective can be applied to understanding ethical relations within pre-Qin Confucianism. In the pre-Classical period, the absence of scientific explanations for human origins shrouded these origins in mysticism, leading to practices such as totem worship, ancestor worship, and the worship of Heaven. However, through continuous exploration of the Heaven–human relationship, pre-Qin Confucians began to liberate human rights from divine authority, highlighting a profound sense of humanism. In light of this transformation, this paper aims to address three key questions: first, what distinctions did pre-Qin Confucians draw between humans and animals, and how did they emphasize individual human value? Second, how did pre-Qin Confucians transform ancestor worship through the traditions of Li and Yue? Third, how did the focus of pre-Qin Confucians shift from Heaven back to humanity? These three research areas are crucial for analyzing how the ethical relations of pre-Qin Confucianism closely integrated with political order. This exploration not only illuminates the philosophical evolution of the period but also offers a deeper understanding of how early Confucianism laid the groundwork for the interplay between personal value and societal responsibilities.

Gentiana macrophylla is a traditional Chinese herbal medicine for treating jaundice, hepatitis, and stomachic and choleretic ailments, and its main secondary metabolites are secoiridiod glycosides represented by gentiopicroside. The biosynthetic pathway of gentiopicroside is via methylerythritol phosphate pathway (MEP) and mevalonate pathway (MVA) pathways. Fosmidomycin and lovastatin are specific inhibitors of 1-deoxy-D-xylulose-5-phosphate reductoisomerase (DXR) of the MEP pathway and hydroxymethylglutaryl-CoA reductase (HMGR) of the MVA pathway, and are widely used in the research of the biosynthesis pathway and regulation mechanism of secondary metabolites. The article aims to test and search for the optimal condition for decreasing the contents of four secoiridiod glycosides compounds (loganic acid, sweroside, swertiamarin and gentiopicroside) in G. macrophylla by using the orthogonal test designs. In this study, the concentration of fosmidomycin(A), concentration of lovastatin(B) and sampling days (C) were taken as three factors, and each factor was taken as four levels to design an orthogonal test, with 16 groups of treatment. HPLC was used to determine the contents of four secoiridiod glycosides, and the data were statistically analyzed. The results were as follows: (1) The contents of the four secoiridiod glycosides compounds in G. macrophylla were most affected by sampling time, followed by fosmidomycin concentration and then lovastatin concentration. (2) After treatment with the optimal inhibitory conditions, the contents of loganic acid, swertiamarin, gentiopicroside and sweroside decreased by 69%, 36%, 33% and 4% respectively. In conclusion, optimal inhibition conditions are determined to be fosmidomycin 400 μmol·L-1, lovastatin 50 μmol·L-1, samples are harvested on six days, which provides the reference for further research on the regulatory mechanism of MEP and MVA pathways in the metabolism and synthesis of secoiridiod glycosides.

ObjectivesThe aim of this article was to compare the differences between Intergrowth-21st (IG-21) and Fenton growth standards in the classification of intrauterine and extrauterine growth restriction (EUGR) in eastern Chinese preterm infants, and detect which one can better relate to neonatal diseases and predict the physical growth outcomes at 3–5 years old.MethodsPremature infants admitted to a tertiary pediatric hospital in Shanghai, China, from 2016 to 2018 were enrolled. Prenatal information, neonatal diseases during hospitalization, and anthropometric data (weight, height, and head circumference) at birth and at discharge were collected and analyzed. Physical growth outcomes (short stature, thinness, and overweight) were examined by telephone investigations in 2021 at age 3–5 years.ResultsThe medium gestational age and birth weight of the included 1,065 preterm newborns were 33.6 weeks and 1,900 g, respectively. The IG-21 curves diagnosed more newborns with small for gestational age (SGA) (19% vs. 14.7%) and fewer newborns with longitudinal EUGR on height (25.5% vs. 27.9%) and head circumference (17.9% vs. 24.7%) compared to Fenton curves. Concordances between Fenton and IG-21 standards were substantial or almost perfect in the classification of SGA and longitudinal EUGR, but minor in cross-sectional EUGR. EUGR identified by Fenton curves was better related to neonatal diseases than IG-21 curves. There were no statistical significances in the prediction of short stature, thinness, and overweight at 3–5 years old between the two charts.ConclusionsIG-21 growth standards are not superior to Fenton in assessing preterm growth and development in the eastern Chinese population.

Abstract Background The primary aim of the study was to assess the nutritional status of pediatric liver transplant outpatients in nutrition clinic, particularly the nutritional status of their bones. Methods One hundred thirty-eight pediatric liver transplant outpatients, who had visited the nutrition clinic in Shanghai Children’s Medical Center between January 2017 and December 2019, were recruited. The bone mineral density (BMD) z-scores were determined by dual energy X-ray absorptiometry (DXA). Nutritional assessment was performed, and their corresponding height-for-age z-scores (HAZs)/weight-for-age z-scores (WAZs)/BMI-for-age z-scores (BMIZs) were obtained. Results A total of 138 patients came to our nutrition outpatient clinic, including 68 boys (49.3%) and 70 girls (50.7%). The median age was 0.87y (0.68y, 1.71y). Among these patients, 44 (31.9%) had acute malnutrition with WAZ/BMIZ value -1.14 (-2.38, -0.18), 55 (38.4%) had chronic malnutrition with HAZ value -1.51 (-2.39, -0.38), and 96 (69.6%) had a BMD lower than normal. The BMD z-score was significantly correlated with the WAZ/BMIZ value (Spearman’s correlation coefficient = 0.334, p

ObjectivesTo investigate the postnatal growth trajectories of preterm infants and evaluate the association between extrauterine growth restriction (EUGR) at discharge and adverse physical growth outcomes at age 3–6 years.MethodsPremature infants admitted to Shanghai Children’s Medical Center within 24 h after birth from 1 January 2016 to 31 December 2018 were enrolled. Neonatal complications, nutrition support, and anthropometric data were collected and analyzed to diagnose EUGR on different definitions at discharge. The weight and the height of each subject were collected by telephone investigation from 1 September 2021 to 31 November 2021 to access the incidences of overweight/obesity, short stature, and thinness at age 3–6 years.ResultsA total of 527 preterm infants were included in the final sample. The overall mean weight and height Z-scores were –0.37 ± 0.97 SD and –0.29 ± 1.18 SD at birth, and increased to –0.03 ± 1.11 SD and 0.13 ± 1.2 SD at follow-up, respectively. The logistic regression analysis indicated longitudinal EUGR on head circumference as the risk factor of overweight or obesity, cross-sectional EUGR on height as the risk factor of short stature, and delayed EN as the risk factor of thinness.ConclusionThe growth trajectories of the preterm newborns tended toward the normal direction. Longitudinal EUGR on the head circumference and cross-sectional EUGR on height at discharge were associated with adverse physical growth outcomes at age 3–6 years.

Abstract Burkholderia pyrrocinia JK-SH007 is an important biocontrol strain for the prevention and treatment of poplar canker disease. Its powerful biocontrol function is inseparable from its successful colonization of poplar trees. Bacterial biofilms can ensure the long-term colonization of a host. To explore the mechanism of action of biofilms in the biocontrol process, we manipulated various exogenous factors to explore the morphology of the JK-SH007 biofilm in vitro. The addition of glycerol and MgSO4 to TSB medium stimulated biofilm production, increased the resistance of JK-SH007 to disease, enhanced the survival of JK-SH007 in nutrient-poor environments and maintained the antagonistic ability of JK-SH007 against the poplar canker pathogen. Therefore, we constructed and optimized a biofilm-forming system to produce a large number of stable JK-SH007 biofilms. The optimized system showed that the optimal incubation time for JK-SH007 biofilm formation was 14 h, the optimal temperature of the static culture was 25 °C, and the optimal pH was 5. The optimal medium for biofilm formation was TSB medium, 1% glycerol and 50 mM MgSO4. RT-qPCR experiments showed that an increase in the expression of the suhB gene promoted JK-SH007 biofilm formation, while an increase in the expression level of the ropN gene inhibited JK-SH007 biofilm formation. The possible mechanism by which JK-SH007 was inhibited by biofilm formation under natural culture was revealed. These results indicate the importance of adding nutrients to JK-SH007 biocides produced on a commercial scale. This is the first report of JK-SH007 producing a long-lasting biofilm that guarantees antagonism.

Dihydroflavonol 4-reductase (DFR), a key enzyme involved in the biosynthesis of anthocyanins, has been cloned from various species. However, little research has been conducted on this enzyme in ferns, which occupy a unique evolutionary position. In this study, we isolated two novel DFR genes from the fern Dryopteris erythrosora. In vitro enzymatic analysis revealed that DeDFR1 and DeDFR2 enzymes can catalyze dihydrokaempferol and dihydroquercetin but cannot catalyze dihydromyricetin. Amino acid sequence analysis showed that DeDFR1 and DeDFR2 have an arginine at the same substrate-specificity-determining site as that in the ferns Salvinia cucullata and Azolla filiculoides. Thus, we speculate that the Arg-type DFR is a new DFR functional type. To further verify the substrate preferences of the Arg-type DFR, an amino acid substitution assay was conducted. When N133 was mutated to R133, Arabidopsis DFR protein completely lost its catalytic activity for dihydromyricetin, as observed for DeDFR1 and DeDFR2. Additionally, heterologous expression of DeDFR2 in the Arabidopsis tt3-1 mutant resulted in increasing anthocyanin accumulation. In summary, DeDFR1 and DeDFR2 are considered to be a new type of DFR with unique structures and functions. The discovery of the Arg-type DFR provides new insights into the anthocyanin biosynthesis pathway in ferns.

Ceratopteris thalictroides, a model fern, has two kinds of gametophytes with different sex expression: male and hermaphrodite. Hermaphroditic gametophytes have one or several archegonia beneath the growing point and a few antheridia at the base or margin. Male gametophytes show a spoon-like shape with much longer than the width and produce many antheridia at the margin and surface. The results of chlorophyll fluorescence detection showed that the photochemical efficiency of hermaphrodites was higher than that of males. By using two-dimensional electrophoresis and mass spectrometry, the differentially abundant proteins in hermaphroditic and male gametophytes were identified. A total of 1136 ± 55 protein spots were detected in Coomassie-stained gels of proteins from hermaphroditic gametophytes, and 1130 ± 65 spots were detected in gels of proteins from male gametophytes. After annotation, 33 spots representing differentially abundant proteins were identified. Among these, proteins involved in photosynthesis and chaperone proteins were over-represented in hermaphrodites, whereas several proteins involved in metabolism were increased in male gametophytes in order to maintain their development under relatively nutritionally deficient conditions. Furthermore, the differentially abundant cytoskeletal proteins detected in this study, such as centrin and actin, may be involved in the formation of sexual organs and are directly related to sex expression. These differentially abundant proteins are important for maintaining the development of gametophytes of different sexes in C. thalictroides.

Manganese (Mn) exposure can affect brain development. Whether Selenium (Se) can protect neonates against neurotoxicity from Mn exposure remains unclear. We investigated this issue in 933 mother-newborn pairs in Shanghai, China, from 2008 through 2009. Umbilical cord serum concentrations of Mn and Se were measured and Neonatal Behavioral Neurological Assessment (NBNA) tests were conducted. The scores

Three furan derivatives (1-3) were isolated from the methanol extract of Gentiana macrophylla Pall. (Gentianaceae). Their structures were determined on the basis of spectroscopic analysis including HR-MS, 1D and 2D NMR. Genfurtate (1) was identified as a new compound, while, swertianglide (2) was firstly reported in genus Gentiana, and anglelone (3) was firstly reported in the plant G. macrophylla. Accordingly, their biosynthesis pathways transformed from secologanin in G. macrophylla was deduced. [ABSTRACT FROM AUTHOR]