Pyruvate Pathway Research Articles

Preparation and characterization of moringin-loaded chitosan-coated liposomes and their antibacterial activity against Staphylococcus aureus

This study aimed to improve the stability of moringin and clarify the inhibitory mechanisms of moringin-loaded chitosan-coated liposomes (MR-CS-LPs) against Staphylococcus aureus. Optimisation of MR-CS-LPs was conducted using the response surface methodology, and extensive characterization was performed. The anti-bacterial activity of MR-CS-LPs was assessed by determining the minimum inhibitory concentration (MIC) and conducting growth curve analyses. The effects of MR-CS-LPs on S. aureus cell wall and membrane integrity were investigated using techniques such as scanning electron microscopy and physical and chemical analyses. Apoptotic effects were evaluated by examining oxidative stress parameters, and the impact on S. aureus biofilm formation was explored. An LC–MS/MS analysis provided insights into the inhibitory mechanism of MR-CS-LPs against S. aureus. The results indicated that MR-CS-LPs achieved an encapsulation rate of 69.02 %. Furthermore, they demonstrated potent anti-bacterial activity against S. aureus, with an MIC of 0.125 mg/mL. MR-CS-LPs disrupted cell wall and membrane integrity, resulting in macromolecule leakage, induced oxidative stress–mediated apoptosis and effectively suppressed biofilm formation, ultimately leading to bacterial death. Metabolomics analysis revealed that MR-CS-LPs inhibit S. aureus by regulating pyruvate pathways. These findings affirm that MR-CS-LPs possess significant anti-microbial properties, underscoring their potential as effective anti-microbial agents against S. aureus.

Efficient carbon flux allocation towards D-pantothenic acid production via growth-decoupled strategy in Escherichia coli

For industrial strain construction, rational allocation of carbon flux is of paramount importance especially for decoupling cell growth and chemical productions to get maximum titer, rate, yield (TRY), which become Gordian Knot. Here, a temperature-sensitive switch and genetic circuits was used for effectively decoupling cell growth from D-pantothenic acid (DPA) production, along with systematically metabolic engineering including blocking redundant pathways of pyruvate and enhancing DPA driving force. Afterwards, rapid biomass accumulation only happened during growth stage, and subsequent high-efficient DPA production was initiated with reducing fermentation temperature. Finally, 97.20 g/L DPA and 0.64 g/g glucose conversion rate were achieved in 5-liter fed-batch fermentation. These undisputedly represent a milestone for the biosynthesis of DPA. With using strategies for decoupling cell growth from chemical productions, it would serve as “Alexander’s sword” to cut Gordian Knot to get industrial chassis cells with excellent TRY for de novo biosynthesis of valuable chemicals.

Antibiotic fermentation residue for biohydrogen production: Inhibitory mechanisms of the inherent antibiotic

Antibiotic fermentation residue, which is generated from the microbial antibiotic production process, has been a troublesome waste faced by the pharmaceutical industry. Dark fermentation is a potential technology to treat antibiotic fermentation residue in terms of renewable H2 generation and waste management. However, the inherent antibiotic in antibiotic fermentation residue may inhibit its dark fermentation performance, and current understanding on this topic is limited. This investigation examined the impact of the inherent antibiotic on the dark H2 fermentation of Cephalosporin C (CEPC) fermentation residue, and explored the mechanisms from the perspectives of bacterial communities and functional genes. It was found that CEP-C in the antibiotic fermentation residue significantly inhibited the H2 production, with the H2 yield decreasing from 17.2 mL/g-VSadded to 12.5 and 9.6 mL/g-VSadded at CEP-C concentrations of 100 and 200 mg/L, respectively. CEP-C also prolonged the H2-producing lag period. Microbiological analysis indicated that CEP-C remarkably decreased the abundances of high-yielding H2-producing bacteria, as well as downregulated the genes involved in hydrogen generation from the“pyruvate pathway” and“NADH pathway”, essentially leading to the decline of H2 productivity. The present work gains insights into how cephalosporin antibiotics influence the dark H2 fermentation, and provide guidance for mitigating the inhibitory effects.

L-tryptophan anaerobic fermentation for indole acetic acid production: Bacterial enrichment and effects of zero valent iron

Indole acetic acid (IAA) as a plant hormone, was one of the valuable products of anaerobic fermentation. However, the enriching method remained unknown. Moreover, whether zero valent iron (ZVI) could enhance IAA production was unexplored. In this work, IAA producing bacteria Klebsiella (63 %) was enriched successfully. IAA average production rate and concentration were up to 3 mg/L/h and 56 mg/L. With addition of 1 g/L ZVI, IAA average production rate and concentration was increased for 2 and 3 folds. Mechanisms indicated ZVI increased Na+K+-ATP activity and electron transport activity for 2 folds and 1 fold. Moreover, macro transcription determined indole pyruvate pathway activity like primary-amine oxidase, indole pyruvate decarboxylase and aldehyde dehydrogenase were increased for 146 %, 187 %, and 557 %, respectively. Therefore, ZVI was suitable for enhancement IAA production from mixed culture anaerobic fermentation.

Profiling the neuroproteomics of honeybee brain: A clue for understanding the role of neuropeptides in the modulation of aggressivity

The aggressivity is modulated in honeybee brain through a series of actions in cascade mode, with the participation of the neuropeptides AmAST A (59–76) and AmTRP (254–262). The aggressivity of honeybees was stimulated by injecting both neuropeptides in the hemocoel of the worker honeybees, which were submitted to behavioral assays of aggression. The brain of stinger individuals were removed by dissection and submitted to proteomic analysis; shotgun proteomic approach of honeybee brain revealed that both neuropeptides activate a series of biochemical processes responsible by production of energy, neuronal plasticity and cell protection. In addition to this, AmTRP (254–262) elicited the expression of proteins related to the processing of the potential of action and lipid metabolism; meanwhile AmAST A (59–76) elicited the metabolism of steroids and Juvenile hormone-related metabolism, amongst others. Apparently, the most complex biochemical process seems to be the regulation of ATP production, which occurs at two levels: i) by a subgroup of proteins common to the three experimental groups, which are over−/under-regulated through glycolysis, pyruvate pathway, Krebbs cycle and oxidative phosphorylation; ii) by a subgroup of proteins unique to the each experimental group, which seems to be regulated through Protein-Protein Interactions, where the protein network regulated by AmTRP (254–262) seems to be more complex than the other two experimental groups. SignificanceRecently we reported the effect of the neuropeptides AmAST A (59–76) and AmTRP (254–262) in the modulation of the aggressive behavior of the worker honeybees. Up to now it is known that the simple presence of the allatostatin and tachykinin-related-peptide in bee brain, is enough for inducing the aggressive behavior. However, nothing was known about how these neuropeptides perform their action, inducing the aggressive behavior. The results of the present study elucidated some of the metabolic pathways that were activated or inhibited to support the complex defensive behavior, which includes the aggressivity. These results certainly will impact the behavioral research of honeybees, since we are paving the way for understanding the molecular base of regulation, of individual /nest defense of honeybees.

In vivo mechanism of the interaction between trimethylamine lyase expression and glycolytic pathways.

Recent studies confirmed that host-gut microbiota interactions modulate disease-linked metabolite TMA production via TMA lyase. However, microbial enzyme production mechanisms remain unclear. In the present study, we investigated the impact of dietary and intervention factors on gut microbiota, microbial gene expression, and the interplay between TMA lyase and glycolytic pathways in mice. Using 16S rRNA gene sequencing, metagenomics, and metabolomics, the gut microbiota composition and microbial functional gene expression profiles related to TMA lyase and glycolytic enzymes were determined. The results revealed that distinct diets and intervention factors altered gut microbiota, gene expression, and metabolites linked to glycine metabolism and glycolysis. Notably, an arabinoxylan-rich diet suppressed genes linked to choline, glycine, glycolysis, and TMA lyase, favoring glycine utilization via pyruvate pathways. Glycolytic inhibitors amplified these effects, mainly inhibiting pyruvate kinase. Our findings underscored the crosstalk between TMA lyase and glycolytic pathways, regulating glycine levels, and suggested avenues for targeted interventions and personalized diets to curb choline TMA lyase production.

Investigation of Urine Organic Acid Profile in Coronavirus Disease (COVID-19) Patients.

Coronavirus disease 2019 (COVID-19) is a viral infection caused by severe acute respiratory syn-drome coronavirus 2 (SARS-CoV-2). The high mortality and morbidity rate and its course with a wide variety of clinical symptoms indicate that the disease affects many metabolic pathways. The aim of our study was to investi-gate the metabolic effects of SARS-CoV-2 by evaluating the urinary organic acid profile in patients with SARS-CoV-2 infection. Can metabolites in urine guide the diagnosis, follow-up, treatment, and prognosis of COVID-19? Forty-two patients, including 30 SARS-CoV-2 RT-PCR positive patients and 12 SARS-CoV-2 RT-PCR negative controls, were studied. SARS-COV-2 RT-PCR of nasopharyngeal swab samples was studied. Urines were evaluated in the GCMS-QP2010 SE Gas Chromatograph Mass Spectrometer (SHIMADZU) device for organic acid profile in the metabolism laboratory. Urine organic acid profile was evaluated by studying 117 organic acids in each patient. Tiglylglycine, 2-hydroxybutyric acid, 4-hydroxyphenylpyruvic acid, 3-hydroxypropionic acid, erythro-4,5-dihydroxyhexanoic acid lactone, 2-hydroxyphenylacetic acid, N-acetyltyrosine, 3-phenyllactic acid, 5-hydroxyindolacetic acid, 3-hydroxysebacic acid, palmitic acid, 3-methylglutaconic acid, 3-methylglutaric acid, lactic acid, pyruvic acid-oxime, 3-hydroxysobutyric acid, and organic acids were found to be increased, compared to the control group. Tiglylglycine has been specifically identified as a potential biomarker of respiratory chain disorders. The deterioration in lipid metabolism and pyruvate pathway in COVID-19 patients was evaluated as remarkable.

Natural product fargesin interferes with H3 histone lactylation via targeting PKM2 to inhibit non-small cell lung cancer tumorigenesis.

Non-small cell lung cancer (NSCLC) is one of the most common malignant tumors. There is an urgent need to find more effective drugs that inhibit NSCLC. Fargesin (FGS) has demonstrated anti-tumor effects; however, its efficacy and the molecular mechanism of inhibiting NSCLC are unclear. Herein, we investigated FGS' inhibitory effects on NSCLC by CCK8 and EdU assays and cell cycle analysis of A549 cells in vitro and in a nude mouse tumor transplantation model in vivo. FGS (10-50 μM) significantly inhibited cell proliferation and down-regulated expression levels of CDK1 and CCND1. Transcriptomic analysis showed that FGS regulated the cell metabolic process pathway. Differential metabolites with FGS treatment were enriched in glycolysis and pyruvate pathways. Cell metabolism assay were used to evaluate the oxygen consumption rate (OCR), Extracellular acidification rate (ECAR) in A549 cells. FGS also inhibited the production of cellular lactate and the expression of LDHA, LDHB, PKM2, and SLC2A1. These genes were identified as important oncogenes in lung cancer, and their binding to FGS was confirmed by molecular docking simulation. Notably, the over-expression and gene silencing experiments signified PKM2 as the molecular target of FGS for anti-tumorigenesis. Moreover, the H3 histone lactylation, were correlated with tumorigenesis, were inhibited with FGS treatment. Conclusively, FGS inhibited the aerobic glycolytic and H3 histone lactylation signaling pathways in A549 NSCLC cells by targeting PKM2. These findings provide evidence of the therapeutic potential of FGS in NSCLC.

Study on the relationship between flavor components and quality of ice wine during freezing and brewing of 'beibinghong' grapes

Ice wine has prominent fruity sweetness and unique, rich aroma compared to wine. The sweetness was accumulating, the acidity and astringency tended to soften of grape berry during the freezing period. The process gave the ice wine balanced taste, with prominent honey sweetness, accompanied by refreshing alcoholic taste, soft acidity and astringency. Eleven key aroma compounds were identified in ice wine through GC–MS and ROAV values. The key aroma compounds were analyzed with Pearson correlation coefficient and fragrance mechanism were speculated. Ethyl acetate and 1-octen-3-ol derived from the aroma of grape, are produced by anaerobic metabolism and lipoxygenase pathways of pyruvate and linoleic acid, respectively. Ester aromas, 2-phenylethanol and 2-methylbutanal were derived from the brewing process, were produced by octanoic acid, caproic acid, phenylalanine and isoleucine through lipid metabolism, Ehrlich pathway and Strecker pathway, respectively. Proposed corresponding control methods based on factors that affect the formation of ice wine aromas.

Promoter methylation as a potential cause for the decrease in pyruvate metabolism during environmentally induced aestivation in Apostichopus japonicus

Apostichopus japonicus is an important aquaculture species with significant economic value. As water temperatures increase during the summer, A. japonicus enters aestivation to resist adverse environments and maintain survival. The most significant feature of aestivation in sea cucumbers is metabolic depression, but the molecular mechanisms involved in this process are not fully explained. In this study, we analyzed the methylation status and gene expression of intestinal tissues from A. japonicus during aestivation, to reveal the mechanism of metabolic depression involved during aestivation from the perspective of epigenetic modifications. Of the 2886 hypermethylated genes and 1534 hypomethylated genes obtained, methylation and transcriptome correlation analysis identified 207 methylation-driven low-expression candidate genes and 71 methylation-driven highly expressed candidate genes. Analysis of the methylation location showed that 5% of differential methylation occurred in the promoter region; those genes that were differentially methylated in the promoter region were also significantly enriched in the pyruvate metabolic pathway. Metabolite content analysis of the glycolytic pathway and the tricarboxylic acid cycle pathway, key sources and consumption pathways of pyruvate, respectively, showed that the content of these metabolites was significantly reduced, suggesting pyruvate metabolic depression of A. japonicus during aestivation. In conclusion, these results reveal that promoter methylation is likely a potential cause of pyruvate metabolic depression during aestivation in A. japonicus. They also provide new insights into the epigenetic regulation mechanism of hypometabolism during aestivation.

Suppression of hnRNP A1 binding to HK1 RNA leads to glycolytic dysfunction in Alzheimer's disease models.

To investigate the mechanism of RNA-binding protein hnRNP A1 in mouse hippocampal neurons (HT22) on glycolysis. RIP and CLIP-qPCR were performed by HT22 in vitro to observe the mechanism of hnRNP A1 regulating the expression of key proteins in glycolysis. The RNA binding domain of hnRNP A1 protein in HT22 was inhibited by VPC-80051, and the effect of hnRNP A1 on glycolysis of HT22 was observed. Lentivirus overexpression of hnRNP A1 was used to observe the effect of overexpression of hnRNP A1 on glycolysis of Aβ25-35-injured HT22. The expression of hnRNP A1 in brain tissues of wild-type mice and triple-transgenic (APP/PS1/Tau) AD mice at different ages was studied by Western blot assay. The results of RIP experiment showed that hnRNP A1 and HK1 mRNA were significantly bound. The results of CLIP-qPCR showed that hnRNP A1 directly bound to the 2605-2821 region of HK1 mRNA. hnRNP A1 inhibitor can down-regulate the expression of HK1 mRNA and HK1 protein in HT22 cells. Overexpression of hnRNP A1 can significantly reduce the toxic effect of Aβ25-35 on neurons via the hnRNP A1/HK1/ pyruvate pathway. In addition, inhibition of hnRNP A1 binding to amyloid precursor protein (APP) RNA was found to increase Aβ expression, while Aβ25-35 also down-regulated hnRNP A1 expression by enhancing phosphorylation of p38 MAPK in HT22. They interact to form bidirectional regulation, further down-regulating the expression of hnRNP A1, and ultimately aggravating glycolytic dysfunction. Protein immunoblotting showed that hnRNP A1 decreased with age in mouse brain tissue, and the decrease was greater in AD mice, suggesting that the decrease of hnRNP A1 may be a predisposed factor in the pathogenesis of AD.

Bacteria pyruvate metabolism modulates AFB1 toxicity in Caenorhabditis elegans

Aflatoxin B1 (AFB1), the most potent mycotoxin and Group 1 human carcinogen, continues to pose a significant public health burden, particularly in developing countries. Increasing evidence has shown the gut microbiota as a key mediator of AFB1 toxicity through multiple interactive host-microbiota activities. In our previous study we observed that disturbances in bacterial pyruvate metabolism might have a significant impact on AFB1 in the host. To further investigate the impact of the pyruvate pathway on AFB1 toxicity in C. elegans, we engineered two bacterial strains (triple-overexpressed and triple-knockout strains with aceB, lpd, and pflB). Additionally, we employed two mutant worm strains (pyk-1 and pdha-1 mutants) known to affect pyruvate metabolism. Our results revealed that the co-metabolism of pyruvate by the host and bacterial strains synergistically influences AFB1 toxicity. Remarkable, we found that bacterial pyruvate metabolism, rather than that of the host, plays a pivotal role in modulating AFB1 toxicity in C. elegans. Our study sheds light on the role of gut microbiota involved in pyruvate metabolism in influencing AFB1 toxicity in C. elegans.

1-methylcyclopropene treatment improves chilling tolerance by regulating IAA biosynthesis, auxin signaling transduction and cell wall degradation in peach fruit

Chilling injury (CI) is a major issue in peach fruit during cold storage, which can be alleviated by 1-methylcyclopropene (1-MCP) treatment. Previous works have focused on ethylene regualtion. Recently, auxin signals were found to play vital roles in both 1-MCP-regulated fruit ripening and cold resistance. However, the information regarding the relationship between auxin metabolism and 1-MCP-induced cold tolerance was still limited, especially in cold-sensitive fruit. To investigate the underlying mechanisms, we performed transcriptome analysis and found that 1-MCP treatment significantly regulated several metabolic pathways, including auxin signaling, cell wall pectin degradation, and indole pyruvate metabolism. This regulation contribute to the development of cold resistance by maintaining cell wall structural integrity through delaying the increase of water-soluble pectin (WSP) and the decrease of CDTA-soluble pectin (CSP) and sodium carbonate-soluble pectin (NSP). Accordingly, 1-MCP treatment reduced the activities of polygalacturonase (PG), pectin methylesterase (PME) and pectin lyase (PEL), and down-regulated the expressions of PpPG1, PpPG2, PpPME1, PpPME2, PpPEL1 and PpPEL2. Moreover, the treatment decreased indole-3-acetic acid (IAA) biosynthesis via the indole pyruvate pathway, which led to the attenuation of the auxin signaling pathway through down-regulation of auxin response factors (PpARF1), auxin-responsive genes (PpAUX/IAA1, PpSAUR1 and PpGH3–1), and auxin receptor proteins (PpTIR1) expressions. Our results suggested 1-MCP treatment improved chilling tolerance by regulating IAA biosynthesis, auxin signaling transduction and cell wall degradation, and provided new insight into the mechanisms of 1-MCP-induced cold resistance in postharvest peach fruit.

Improved Assessment of Soil Nonextractable Residues of the Pyrethroid Insecticide Cyphenothrin.

The metabolic fate of pyrethroid insecticide cyphenothrin (1) [(RS)-α-cyano-3-phenoxybenzyl (1RS)-cis-trans-2,2-dimethyl-3-(2-methylprop-1-enyl)cyclopropanecarboxylate] in soils was investigated using 14C-labeled (1R)-cis/trans isomers at the cyclopropane ring. Both isomers degraded with half-lives of 19.0-47.4 days, and 48.9-56.0% and 27.5-38.7% of the applied radioactivity (AR) were mineralized to CO2 and incorporated into nonextractable residues (NER), respectively, after 120 days at 20 °C. NER analyses revealed 37.5-42.2% (cis-1) and 44.9-54.1% (trans-1) of each residue at 30/120 days were comprised of 14C-amino acids (AAs) as microbial products. Assuming that 50% of microbial biomass is AAs, it was estimated that 11.3-22.9%AR (cis-1, 75.0-84.4% of NER) and 13.9-30.4%AR (trans-1, 89.8-108.2% of NER) were nonhazardous biogenic NER (bio-NER), while type I/II xenobiotic NER (xeno-NER) characterized by silylation was insignificant at 0.9-1.0%/2.8-3.3%AR (cis-1). Detailed 14C-AA quantitation indicated a high relevance of the tricarboxylic acid cycle and pyruvate pathway during bio-NER formation, offering new insights into the microbial assimilation of the chrysanthemic moiety.

Host-microbiota affects the toxicity of Aflatoxin B1 in Caenorhabditis elegans.

Aflatoxins are a group of potent fungal metabolites produced by Aspergillus and commonly contaminate groundnuts and cereal grains. Aflatoxin B1 (AFB1), the most potent mycotoxin, has been classified as Group 1 human carcinogen because it can be metabolically activated by the cytochrome P450 (CYP450) in the liver to form AFB1-DNA adducts and induce gene mutations. Increasing evidence has shown the gut microbiota as a key mediator of AFB1 toxicity through multiple interactive host-microbiota activities. To identify specific bacterial activity that modulates AFB1 toxicity in Caenorhabditis (C.) elegans, we established a 3-way (microbe-worm-chemical) high-throughput screening system using C. elegans fed E. coli Keio collection on an integrated robotic platform, COPAS Biosort. We performed 2-step screenings using 3985 Keio mutants and identified 73 E. coli mutants that modulated C. elegans growth phenotype. Four genes (aceA, aceB, lpd, and pflB) involved in the pyruvate pathway were identified from the screening and confirmed to increase the sensitivity of all animals to AFB1. Taking together, our results indicated that disturbances in bacterial pyruvate metabolism might have a significant impact on AFB1 toxicity in the host.

A new capacity of gut microbiota: Fermentation of engineered inorganic carbon nanomaterials into endogenous organic metabolites

Carbon-based nanomaterials (CNMs) have recently been found in humans raising a great concern over their adverse roles in the hosts. However, our knowledge of the invivo behavior and fate of CNMs, especially their biological processes elicited by the gut microbiota, remains poor. Here, we uncovered the integration of CNMs (single-walled carbon nanotubes and graphene oxide) into the endogenous carbon flow through degradation and fermentation, mediated by the gut microbiota of mice using isotope tracing and gene sequencing. As a newly available carbon source for the gut microbiota, microbial fermentation leads to the incorporation of inorganic carbon from the CNMs into organic butyrate through the pyruvate pathway. Furthermore, the butyrate-producing bacteria are identified to show a preference for the CNMs as their favorable source, and excessive butyrate derived from microbial CNMs fermentation further impacts on the function (proliferation and differentiation) of intestinal stem cells in mouse and intestinal organoid models. Collectively, our results unlock the unknown fermentation processes of CNMs in the gut of hosts and underscore an urgent need for assessing the transformation of CNMs and their health risk via the gut-centric physiological and anatomical pathways.

Erchen decoction to reduce oxidative stress in dyslipidemia phlegm-dampness retention syndrome mice: In vivo mechanism revealed by metabolomics (liquid chromatography–mass spectrometry)

ObjectiveErchen decoction, a traditional Chinese medicine formula, can reduce the level of oxidative stress for the treatment of dyslipidemia phlegm-dampness retention syndrome (DPDRS); however, studies have not elucidated the mechanism underlying its metabolic action. Here, liquid chromatography–mass spectrometry (LC–MS)-based metabolomic techniques were utilized to characterize the in vivo effects of Erchen decoction in achieving reduction of oxidative stress levels and understand the potential metabolic mechanisms of action. MethodsWe constructed a DPDRS animal model using a multifactorial composite modeling approach, and Erchen decoction was administered by gavage. We employed LC–MS-based metabolomic techniques in combination with serum-associated factors, gene transcription, methylation detection, and hematoxylin and eosin staining. ResultsIn this study, the constructed animal model of DPDRS had satisfactory quality. Erchen decoction treatment reduced the levels of low-density lipoprotein cholesterol, t total cholesterol and riglyceride; it improved the endothelial structure, increased levels of serum β-nicotinamide adenine dinucleotide phosphate and glutathione concentrations, increased aortic phosphoserine aminotransferase and phosphoserine phosphatase gene expression levels, and decreased aortic phosphoglycerate dehydrogenase methylation level. A total of 64 differential metabolites were obtained using LC–MS assay, and 34 differential metabolic pathways were obtained after enrichment. ConclusionsErchen decoction treatment of DPDRS mice reversed lipid indexes, improved vascular endothelial structure, increased serum and aortic anti-oxidative stress factor concentration and expression levels, and decreased methylation levels, thereby reducing oxidative stress and protecting vascular endothelium. Tricarboxylic acid cycle and metabolic pathways of serum glutamine, serine, tryptophan, pyrimidine, and pyruvate were the most relevant metabolic pathways involved in reducing oxidative stress levels by Erchen decoction during DPDRS treatment; especially, mitochondrial redox homeostasis maintenance in endothelial cells may be crucial. In this work, the therapeutic potential of Erchen decoction for reducing the oxidative stress level in DPDRS was demonstrated; however, its in-depth mechanism is worth further exploration.

Pharmacological activities and effective substances of the component-based Chinese medicine of Ginkgo biloba leaves based on serum pharmacochemistry, metabonomics and network pharmacology.

As a potential drug candidate for the treatment of hypertension and complications, it is speculated that the component-based Chinese medicine of Ginkgo biloba leaves (GBCCM) which mainly composed of flavonoid aglycones (FAs) and terpene lactones (TLs) may have different pharmacological effects at different doses or ratios. Taking the normal mice as the study object, metabonomics was conducted by giving different doses of GBCCM. Based on the components of GBCCM absorbed into the blood, the network pharmacological prediction was carried out. By integrating the results of metabonomics and network pharmacology, predict the possible pharmacological effects of GBCCM and conduct experimental verification. It was found that eight of the 19 compounds in GBCCM could be absorbed into the blood. GBCCM mainly affected the signal pathways of unsaturated fatty acid, pyruvate, bile acid, melanin and stem cells. It was speculated that GBCCM might have activities such as lowering blood pressure, regulating stem cell proliferation and melanogenesis. By establishing the models of mushroom tyrosinase, rat bone marrow mesenchymal stem cells (BMSCs) and spontaneously hypertensive rats (SHRs), we found that FAs and TLs showed synergistic effect in hypertension and tyrosinase models, and the optimal ratio was 3:2 (4.4mg/kg) and 1:1 (0.4mg/ml), respectively. As effective substances, FAs significantly promoted the proliferation of rat BMSCs on the third and fifth days at the concentration of 0.2μg/ml (p < 0.05). GBCCM showed a variety of pharmacological effects at different doses and ratios, which provided an important reference for the druggability of GBCCM.

Transcriptome and Metabolome Analyses Reveal New Insights into the Regulatory Mechanism of Head Milled Rice Rate.

The head milled rice rate (HMRR) is the most important trait of milling quality, which affects the final yield and quality of rice. However, few genes related to HMRR have been identified and the regulatory mechanism of HMRR remains elusive. In this study, we performed a comparative analysis integrating the transcriptome sequencing of developing seeds at the grain-filling stage and a metabolome analysis of brown rice between two groups of accessions with contrasting performances in HMRR. A total of 768 differentially expressed genes (DEGs) were identified between the transcriptome profiles of low-HMRR and high-HMRR accessions. In comparison to the high-HMRR accessions, 655 DEGs were up-regulated in the low-HMRR accessions, which was 4.79 folds higher than the number of down-regulated genes. These up-regulated DEGs were enriched in various metabolic and biosynthetic processes, oxidation reduction, phosphorylation, ion transport and ATP-related processes. However, the 113 down-regulated DEGs in the low-HMRR accessions were concentrated in carbohydrate metabolic processes, cell-death-related processes and defense response. Among the 30 differential metabolites, 20 and 10 metabolites were down-/up-regulated, respectively, in the accessions with low HMRR. In addition, 10 differential metabolites, including five metabolites of the shikimate pathway and five metabolites of the pyruvate pathway, were integrated into two separate pathways, starting from sucrose. Our global analysis of HMRR provides an invaluable resource for a better understanding of the molecular mechanism underlying the genetic regulation of HMRR.

Roles of Fe-C amendment on sulfate-containing pharmaceutical wastewater anaerobic treatment: Microbial community and sulfur metabolism.

The effects of multiple two-phase anaerobic treatment involving acidification coupling Fe-C on sulfate-containing chemical synthesis-based pharmaceutical wastewater treatment were investigated. Fe-C was added as a filler with 25% vol. to acidogenic reactors for semi-continuous operation. The results suggested that Fe-C amendment promoted sulfate removal efficiency by 47.5% and shortened the reaction time by 50% in the acidogenic phase. With mitigation of sulfate inhibition, SCOD removal efficiency and methane production were further increased by 24.6% and 398% compared to direct raw wastewater anaerobic digestion, respectively, in methanogenic phase. The results of sulfate removal kinetics confirmed a 150% increase of removal rate in acidogenic phase. However, the apparent kinetic microbial sulfate removal constant without Fe-C amendment was maintained at approximately 0.06 h-1. The Fe-C amendment not only increased the relative abundance of Methanothrix and Desulfovibrio for sulfate reduction but also enriched unclassified_p__Chloroflexi and unclassified_c__Deltaproteobacteria for acidification. Metagenomic results indicated that Fe-C enhanced dissimilatory sulfate reduction and PAPS synthesis of assimilatory step. The hydrogen sulfide production through the 3-mercaptopyruvate to pyruvate pathways was also enhanced. Butyrate-oxidizing genes were increased synchronously to convert butyrate to acetate.