Untargeted Metabolomic Method Research Articles

Effect of exogenous calcitriol on myopia development and axial length in guinea pigs with form deprivation myopia

The annual increase in myopia prevalence poses a significant economic and health challenge. Our study investigated the effect of calcitriol role in myopia by inducing the condition in guinea pigs through form deprivation for four weeks. Untargeted metabolomics methods were used to analyze the differences in metabolites in the vitreous body, and the expression of vitamin D receptor (VDR) in the retina was detected. Following form deprivation, the guinea pigs received intraperitoneal injections of calcitriol at different concentrations. We assessed myopia progression using diopter measurements and biometric analysis after four weeks. Results indicated that form deprivation led to a pronounced shift towards myopia, characterized by reduced choroidal and scleral thickness, disorganized collagen fibers, and decreased scleral collagen fiber diameter. Notably, a reduction in calcitriol expression in vitreous body, diminished vitamin D and calcitriol levels in the blood, and decreased VDR protein expression in retinal tissues were observed in myopic guinea pigs. Calcitriol administration effectively slowed myopia progression, preserved choroidal and scleral thickness, and prevented the reduction of scleral collagen fiber diameter. Our findings highlight a significant decrease in calcitriol and VDR expressions in myopic guinea pigs and demonstrate that exogenous calcitriol supplementation can halt myopia development, enhancing choroidal and scleral thickness and scleral collagen fiber diameter.

Development of an Untargeted LC-MS Metabolomics Method with Postcolumn Infusion for Matrix Effect Monitoring in Plasma and Feces.

Untargeted metabolomics based on reverse phase LC-MS (RPLC-MS) plays a crucial role in biomarker discovery across physiological and disease states. Standardizing the development process of untargeted methods requires paying attention to critical factors that are under discussed or easily overlooked, such as injection parameters, performance assessment, and matrix effect evaluation. In this study, we developed an untargeted metabolomics method for plasma and fecal samples with the optimization and evaluation of these factors. Our results showed that optimizing the reconstitution solvent and sample injection amount was critical for achieving the balance between metabolites coverage and signal linearity. Method validation with representative stable isotopically labeled standards (SILs) provided insights into the analytical performance evaluation of our method. To tackle the issue of the matrix effect, we implemented a postcolumn infusion (PCI) approach to monitor the overall absolute matrix effect (AME) and relative matrix effect (RME). The monitoring revealed distinct AME and RME profiles in plasma and feces. Comparing RME data obtained for SILs through postextraction spiking with those monitored using PCI compounds demonstrated the comparability of these two methods for RME assessment. Therefore, we applied the PCI approach to predict the RME of 305 target compounds covered in our in-house library and found that targets detected in the negative polarity were more vulnerable to the RME, regardless of the sample matrix. Given the value of this PCI approach in identifying the strengths and weaknesses of our method in terms of the matrix effect, we recommend implementing a PCI approach during method development and applying it routinely in untargeted metabolomics.

Changes of anthocyanin and amino acid metabolites in saffron petals (Crocus sativus L.) during fermentation based on untargeted metabolomics

In order to investigate the possibility of fermentation for deep processing of saffron petals, the metabolic profiles of anthocyanins and amino acids in saffron petals were studied by untargeted metabolomics method based on ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS) combined with multivariate statistical analysis. The PCA, OPLS-DA and HCA models were established to analyze the differential metabolites before and after fermentation. Subsequently, the differential metabolites were screened by OPLS-DA with pairwise modeling. The contents of 26 kinds of anthocyanins during fermentation and 23 kinds of anthocyanins after fermentation were significantly different, respectively. After fermentation, the total anthocyanin content increased, and the monomer anthocyanin produced a variety of glucoside anthocyanin. As for amino acid metabolism, there were significant differences in 24 amino acids during fermentation, 27 kinds of amino acids after fermentation, and then the total content of amino acids decreased. In addition, based on the KEEG database, several metabolic pathways during the fermentation process were identified, and the metabolism pathway of anthocyanin is primarily related to anthocyanin biosynthesis, while the metabolic pathway of amino acid is mainly related to metabolic pathways. These findings provide comprehensive insights into the quality improvements of saffron petals wine.

UHPLC-QTOF-MS-based untargeted metabolomic authentication of Chinese red wines according to their grape varieties

Wine is a very popular alcoholic drink owing to its health benefits of antioxidant effects. However, profits-driven frauds of wine especially false declarations of variety frequently occurred in markets. In this work, an UHPLC-QTOF-MS-based untargeted metabolomics method was developed for metabolite profiling of 119 bottles of Chinese red wines from four varieties (Cabernet Sauvignon, Merlot, Cabernet Gernischt, and Pinot Noir). The metabolites of red wines from different varieties were assessed using orthogonal partial least-squares discriminant analysis (OPLS-DA) and analyzed using KEGG metabolic pathway analysis. Results showed that the differential compounds among different varieties of red wines are mainly flavonoids, phenols, indoles and amino acids. The KEGG metabolic pathway analysis showed that indoles metabolism and flavonoids metabolism are closely related to wine varieties. Based on the differential compounds, OPLS-DA models could identify external validation wine samples with a total correct rate of 90.9 % in positive ionization mode and 100 % in negative ionization mode. This study indicated that the developed untargeted metabolomics method based on UHPLC-QTOF-MS is a potential tool to identify the varieties of Chinese red wines.

Metabolomic profiling of preterm birth in pregnant women living with HIV

BackgroundPreterm birth is a leading cause of death in children under the age of five. The risk of preterm birth is increased by maternal HIV infection as well as by certain antiretroviral regimens, leading to a disproportionate burden on low- and medium-income settings where HIV is most prevalent. Despite decades of research, the mechanisms underlying spontaneous preterm birth, particularly in resource limited areas with high HIV infection rates, are still poorly understood and accurate prediction and therapeutic intervention remain elusive.ObjectivesMetabolomics was utilized to identify profiles of preterm birth among pregnant women living with HIV on two different antiretroviral therapy (ART) regimens.MethodsThis pilot study comprised 100 mother-infant dyads prior to antiretroviral initiation, on zidovudine monotherapy or on protease inhibitor-based antiretroviral therapy. Pregnancies that resulted in preterm births were matched 1:1 with controls by gestational age at time of sample collection. Maternal plasma and blood spots at 23–35 weeks gestation and infant dried blood spots at birth, were assayed using an untargeted metabolomics method. Linear regression and random forests classification models were used to identify shared and treatment-specific markers of preterm birth.ResultsClassification models for preterm birth achieved accuracies of 95.5%, 95.7%, and 80.7% in the untreated, zidovudine monotherapy, and protease inhibitor-based treatment groups, respectively. Urate, methionine sulfone, cortisone, and 17α-hydroxypregnanolone glucuronide were identified as shared markers of preterm birth. Other compounds including hippurate and N-acetyl-1-methylhistidine were found to be significantly altered in a treatment-specific context.ConclusionThis study identified previously known as well as novel metabolomic features of preterm birth in pregnant women living with HIV. Validation of these models in a larger, independent cohort is necessary to ascertain whether they can be utilized to predict preterm birth during a stage of gestation that allows for therapeutic intervention or more effective resource allocation.

THU307 Landscapes Of MaternalAnd Neonatal Bacterial And Metabolic SignaturesAnd Their Interaction In Gestational Diabetes Mellitus

Abstract Disclosure: L. Ding: None. J. Liu: None. X. Zhai: None. X. Xiao: None. Background: Gestational diabetes mellitus (GDM) is strongly correlated with considerable risks to both the mother and the developing fetus. Although evidence indicates an association between the altered gut microbiota and GDM, the microbial and metabolic mechanisms that link the metabolic state between mothers and offspring are unknown. This study aims to characterize the maternal and neonatal bacterial and metabolic signatures and their interaction in gestational diabetes mellitus. Methods: Sixty-six mothers (39 women with GDM and 27 normal pregnant women) and eighty-nine newborns (44 from GDM mothers and 45 from normal mothers) were included in this study. Using 16S rRNA gene sequencing and untargeted metabolomic methods, we determined the gut microbiota and serum metabolomics both for mothers and newborns. Results: We found that the microbial and metabolic signatures of GDM mothers and newborns were significantly different from those of the Control group (Ctr), respectively. Intriguingly, we identified 14 genera (Phyllobacterium, etc.) that exhibited identical alterations in mothers and newborns between groups. Among the overlapped differential metabolites in mothers and newborns, 23 (Glutamyl-methionine, etc.) showed consistent alterations between groups, while 19 (Cyclotetradecane, etc.) metabolites exhibited opposite alterations. Metabolic KEGG pathway analysis revealed that 6 pathways, including fatty acid metabolism, sphingolipid metabolism, TCA cycle, transsulfuration pathway, bile acid metabolism, and carnitine metabolism were both enriched in mothers and newborns. Significantly, correlation analysis revealed the notable association between pyroglutamic acid and maternal glucose level in Ctr group was disrupted in GDM group (Ctr: r = -0.46, p = 0.02; GDM: r = 0.04, p = 0.82). Simultaneously, pyroglutamic acid was significantly correlated with neonate serum high-density lipoprotein cholesterol level, which was disrupted in newborns of GDM mothers (Ctr: r = 0.33, p = 0.03; GDM: r = -0.05, p = 0.80). Conclusion: Altogether, we demonstrated that GDM could consistently alter the microbiota and serum metabolome in mothers and newborns, which is significantly associated with their metabolic state respectively. These findings indicate that serum metabolites serve as promising targets to intervene in metabolic transmission in GDM, which needs further validation. Presentation: Thursday, June 15, 2023

Crotonis Fructus-induced gut microbiota and serum metabolic disorders in rats.

Crotonis Fructus (CF), a poisonous traditional laxative, has been used to treat constipation, edema, ascites, and inflammation for more than 2000years. However, CF possesses toxicity and its toxic mechanism is still unclear. Thus, this research explored the deleterious impacts and underlying mechanisms of CF by evaluating alterations in gut microbiota composition and metabolites. High-throughput sequencing was employed on the 16S rDNA gene to explore the intestinal flora. The untargeted metabolomics method was utilized for evaluating serum metabolomics analysis. The results showed that CF could induce obvious hepatic and gastrointestinal damage by histopathologic morphology of the liver, stomach, duodenum, and colon. According to 16S rDNA sequencing, CF can cause gut microbiota disturbance in rats, and the abundance of opportunistic pathogens such as Clostridia_UCG_014_unclassified increased significantly, while the levels of beneficial bacterial Lactobacillus remarkably declined after CF treatment. Additionally, metabolomics analysis demonstrated that CF may induce toxicity by disrupting the glycerophospholipid metabolism pathway and metabolites such as phosphatidylcholine and phosphatidylethanolamine. Moreover, a correlation study revealed the link between intestinal flora, serum metabolites, and toxicity-related biochemical markers. The results provide a new idea for the research and clinical application of toxic traditional medicine. KEY POINTS: • Crotonis Fructus could affect the gut flora and serum metabolic disruption in SD rats. • Crotonis Fructus could promote the proliferation of harmful bacteria and inhibit beneficial bacteria. • Glycerophospholipid metabolism was disturbed by Crotonis Fructus.

Mitochondrial citrate metabolism and efflux regulate BeWo differentiation

Cytotrophoblasts fuse to form and renew syncytiotrophoblasts necessary to maintain placental health throughout gestation. During cytotrophoblast to syncytiotrophoblast differentiation, cells undergo regulated metabolic and transcriptional reprogramming. Mitochondria play a critical role in differentiation events in cellular systems, thus we hypothesized that mitochondrial metabolism played a central role in trophoblast differentiation. In this work, we employed static and stable isotope tracing untargeted metabolomics methods along with gene expression and histone acetylation studies in an established BeWo cell culture model of trophoblast differentiation. Differentiation was associated with increased abundance of the TCA cycle intermediates citrate and α-ketoglutarate. Citrate was preferentially exported from mitochondria in the undifferentiated state but was retained to a larger extent within mitochondria upon differentiation. Correspondingly, differentiation was associated with decreased expression of the mitochondrial citrate transporter (CIC). CRISPR/Cas9 disruption of the mitochondrial citrate carrier showed that CIC is required for biochemical differentiation of trophoblasts. Loss of CIC resulted in broad alterations in gene expression and histone acetylation. These gene expression changes were partially rescued through acetate supplementation. Taken together, these results highlight a central role for mitochondrial citrate metabolism in orchestrating histone acetylation and gene expression during trophoblast differentiation.

UHPLC-QTOF-MS-based untargeted metabolomics revealing the differential chemical constituents and its application on the geographical origins traceability of lily bulbs

Lily bulbs have been historically used as an edible and medicinal homologous plant. Identifying the geographical origins of lily bulbs produced in specific origin is of great importance since the geographical origins of lily bulbs influence their quality and price greatly. In this work, an untargeted metabolomic method based on UHPLC-QTOF-MS was established for revealing the differential chemical constituents of lily bulbs among different origins and predicting the geographical origins of them by chemometric modeling. A total of 15 differential compounds were screened and identified from untargeted metabolomic data of 50 lily bulb samples by our previously developed AntDAS software. Principal component analysis (PCA) and hierarchical cluster analysis (HCA) showed that samples from five different origins were obviously distinguished based on the differential compounds. What's more, 7 and 6 key characteristic markers were discovered by partial least squares-discriminant analysis (PLS-DA) and orthogonal partial least squares-discriminant analysis (OPLS-DA), respectively. Finally, heatmap, PLS-DA and OPLS-DA models were reconstructed based on the discovered key characteristic markers, and external validation lily bulb samples were successfully discriminated, with recognition rate of 100%. This study demonstrated that the proposed strategy has great potentials for the differentiation and identification of the geographical origins of lily bulbs.

Metabolic profiles and morphological characteristics of leaf tips among different sweet potato (Ipomoea batatas Lam.) varieties

Sweet potato leaf tips have high nutritional value, and exploring the differences in the metabolic profiles of leaf tips among different sweet potato varieties can provide information to improve their qualities. In this study, a UPLC-Q-Exactive Orbitrap/MS-based untargeted metabolomics method was used to evaluate the metabolites in leaf tips of 32 sweet potato varieties. Three varieties with distinct overall metabolic profiles (A01, A02, and A03), two varieties with distinct profiles of phenolic acids (A20 and A18), and three varieties with distinct profiles of flavonoids (A05, A12, and A16) were identified. In addition, a total of 163 and 29 differentially expressed metabolites correlated with the color and leaf shape of sweet potato leaf tips, respectively, were identified through morphological characterization. Group comparison analysis of the phenotypic traits and a metabolite-phenotypic trait correlation analysis indicated that the color differences of sweet potato leaf tips were markedly associated with flavonoids. Also, the level of polyphenols was correlated with the leaf shape of sweet potato leaf tips, with lobed leaf types having higher levels of polyphenols than the entire leaf types. The findings on the metabolic profiles and differentially expressed metabolites associated with the morphology of sweet potato leaf tips can provide useful information for breeding sweet potato varieties with higher nutritional value.

Integrated mass spectrometry imaging and metabolomics reveals sublethal effects of indoxacarb on the red fire ant Solenopsis invicta.

Indoxacarb, representing an efficient insecticide, is normally made into a bait to spread the poison among red fire ants so that it can be widely applied in the prevention and control of Solenopsis invicta. However, the potential toxicity mechanism of S. invicta in response to indoxacarb remains to be explored. In this study, we integrated mass spectrometry imaging (MSI) and untargeted metabolomics methods to reveal disturbed metabolic expression levels and spatial distribution within the whole-body tissue of S. invicta treated with indoxacarb. Metabolomics results showed a significantly altered level of metabolites after indoxacarb treatment, such as carbohydrates, amino acids and pyrimidine and derivatives. Additionally, the spatial distribution and regulation of several crucial metabolites resulting from the metabolic pathway and lipids can be visualized using label-free MSI methods. Specifically, xylitol, aspartate, and uracil were distributed throughout the whole body of S. invicta, while sucrose-6'-phosphate and glycerol were mainly distributed in the abdomen of S. invicta, and thymine was distributed in the head and chest of S. invicta. Taken together, the integrated MSI and metabolomics results indicated that the toxicity mechanism of indoxacarb in S. invicta is closely associated with the disturbance in several key metabolic pathways, such as pyrimidine metabolism, aspartate metabolism, pentose and glucuronate interconversions, and inhibited energy synthesis. Collectively, these findings provide a new perspective for the understanding of toxicity assessment between targeted organisms S. invicta and pesticides. © 2023 Society of Chemical Industry.

Untargeted Metabolomic Characterization of Glioblastoma Intra-Tumor Heterogeneity Using OrbiSIMS.

Glioblastoma (GBM) is an incurable brain cancer with a median survival of less than two years from diagnosis. The standard treatment of GBM is multimodality therapy comprising surgical resection, radiation, and chemotherapy. However, prognosis remains poor, and there is an urgent need for effective anticancer drugs. Since different regions of a single GBM contain multiple cancer subpopulations ("intra-tumor heterogeneity"), this likely accounts for therapy failure as certain cancer cells can escape from immune surveillance and therapeutic threats. Here, we present metabolomic data generated using the Orbitrap secondary ion mass spectrometry (OrbiSIMS) technique to investigate brain tumor metabolism within its highly heterogeneous tumor microenvironment. Our results demonstrate that an OrbiSIMS-based untargeted metabolomics method was able to discriminate morphologically distinct regions (viable, necrotic, and non-cancerous) within single tumors from formalin-fixed paraffin-embedded tissue archives. Specifically, cancer cells from necrotic regions were separated from viable GBM cells based on a set of metabolites including cytosine, phosphate, purine, xanthine, and 8-hydroxy-7-methylguanine. Moreover, we mapped ubiquitous metabolites across necrotic and viable regions into metabolic pathways, which allowed for the discovery of tryptophan metabolism that was likely essential for GBM cellular survival. In summary, this study first demonstrated the capability of OrbiSIMS for in situ investigation of GBM intra-tumor heterogeneity, and the acquired information can potentially help improve our understanding of cancer metabolism and develop new therapies that can effectively target multiple subpopulations within a tumor.

Bile metabolites as diagnostic biomarkers for perihilar cholangiocarcinoma

It is difficult to directly obtain pathological diagnosis of perihilar cholangiocarcinoma (pCCA). Analysis of bile in the pCCA microenvironment, based on metabolomics and statistical methods, can help in clinical diagnosis. Clinical information, bile samples, blood liver function, blood CA199, CEA, and other indicators were collected from 33 patients with pCCA and 16 patients with gallstones. Bile samples were analyzed using untargeted metabolomics methods. A combination of multivariate and univariate analyses were used to screen for potential differential metabolites Through Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment and differential metabolite remodeling, we explored changes in the pCCA pathway and potential therapeutic targets. There were significant differences in patient blood TBIL, ALT, AST, TBA, CA19-9, and CEA indices (p < 0.05, |log2(fc)| ≥ 1) between two groups. A significant correlation was found between these different indicators by Spearman's analysis. The clinical parameters were correlated with mass-to-charge ratios of 305 (Positive Ion Mode, POS) and 246 (Negative Ion Mode, NEG) in the metabolic group (|r| ≥ 0.7, P ≤ 10−7). The result of this study indicated that bile untargeted metabolomics combined with statistical analysis techniques may be used for diagnose and treatment of pCCA.

Positive Effect of High Zinc on Growth of Sedum alfredii

Sedum alfredii Hance (S. alfredii) is a native hyperaccumulator plant species in China that has strong tolerance and accumulation ability for Zn and Cd. In addition, it is a good material for the phytoextraction of soil heavy metal pollutants. However, the specific effect of high Zn concentrations on the growth of S. alfredii and its metabolic mechanisms are not clear. Using an untargeted metabolomics method, we analysed the differential metabolites of the two ecotypes in S. alfredii roots under different Zn treatments. The results showed that high Zn levels significantly promoted plant growth in the hyperaccumulating ecotype (HE), while growth was inhibited in the non-hyperaccumulating ecotype (NHE). We detected 624 metabolites in the roots of S. alfredii. Under the high Zn treatment, lots of lipids and lipid-like molecules, such as glyceryl monooleate and 9,12,13-trihydroxyoctadecane-10-enoic acid, along with organic acids, such as lauramidopropylbetaine, L-malic acid, and their derivatives, decreased significantly in HE roots. Differential metabolites, such as some lipids and lipid-like molecules, were significantly upregulated in NHE roots. The above results indicate that the exogenous high Zn treatment induces the downregulation of HE differential metabolites in response to Zn, but significantly induces the upregulation of differential metabolites in NHE.

Lysophospholipids and branched chain amino acids are associated with aging: a metabolomics-based study of Chinese adults

BackgroundAging is an inevitable process associated with impairments in multiple organ systems, which increases the risk of comorbidity and disability, and reduces the health-span. Metabolomics is a powerful tool in aging research, which can reflect the characteristics of aging at the level of terminal metabolism, and may contribute to the exploration of aging mechanisms and the formulation of anti-aging strategies.MethodsTo identify possible biomarkers and pathways associated with aging using untargeted metabolomics methods, we performed liquid chromatography–mass spectrometry (LC–MS)-based untargeted metabolomics profiling on serum samples from 32 older adults and 32 sex-matched young controls.ResultsMetabolite profiling could distinguish the two groups. Among the 349 metabolites identified, 80—including lysophospholipids whose levels gradually decline—are possible candidate aging biomarkers. Valine, leucine and isoleucine degradation and biosynthesis were important pathways in aging, with reduced levels of l-isoleucine (r = − 0.30, p = 0.017) and l-leucine (r = − 0.32, p = 0.010) observed in older adults.ConclusionsWe preliminarily revealed the metabolite changes associated with aging in Chinese adults. Decreases in mitochondrial membrane-related lysophospholipids and dysfunction of branched-chain amino acid metabolism were determined to be the characteristics and promising research targets for aging.

The grading quality markers identification of Panax notoginseng under the guidance of traditional experience using untargeted metabolomics and anti-myocardial ischemia evaluation of zebrafish

Panax notoginseng (PN) was an edible Chinese herbal medicine. PN's current quality control standard cannot precisely match the traditional grading experience. In this study, under the guidance of the traditional grading experience, the combined metabolomics and biological effect evaluation were used to reveal the distinct chemical quality of PN. The quality of PN was evaluated by traditional experience and characterized by the electronic tongue. A zebrafish myocardial ischemia model was developed to verify the grading experience. The untargeted metabolomics method was used to identify and validate the grading markers of PN. The taste was the critical indicator for classifying the quality. Based on the experience sensory scores (ranged from 47.0 to 87.8), PNs could be divided into two grades. The experience scores were significantly associated with umami and richness of the electronic tongue(p<0.01). Besides, superior PN showed substantially stronger anti-myocardial ischemia activity(p<0.001). Thirty-nine differential components were found using UHPLC-LTQ-Orbitrap MS, of which 22 were identified. A new kind of grading quality markers alkynols in PN-associated efficacy was identified, which revealed stronger anti-myocardial ischemia activities than saponin. This study evaluated PN through untargeted metabolomics and anti-myocardial ischemia evaluation of zebrafish and proposed the critical role of alkynols in PN's quality classification.

Metabolic mechanism of nitrogen modified atmosphere storage on delaying quality deterioration of rice grains.

Nitrogen modified atmosphere was an effective way to control pest infestation in grains. In this study, the quality changes of rice during nitrogen modified atmosphere packaging storage (N2-MAPS) were monitored. An un-targeted metabolomics method was used to detect the rice metabolites and explore the mechanism of N2-MAPS for delaying rice deterioration. In this study, two rice species were studied under N2-MAPS and conventional storage at 30°C for 150days. The quality changes of rice during storage were monitored. The results showed that N2-MAPS could retard the increase of fatty acid value and amylose content, and defer the decrease of enzyme activities. And N2-MAPS had no significant influence on texture characteristics of rice. The metabolomics results suggested some metabolites and pathways were affected by N2-MAPS and revealed that N2-MAPS could protect rice cells from oxidative damage, maintain cell integrity and stability by regulating the metabolism to delay the rice deterioration.

Polarity-extended composition profiling via LC-MS-based metabolomics approaches: A key to functional investigation of Citrus aurantium L

Citrus is a genus containing diverse edible species, among them Citrus aurantium L. is widely utilized while short of composition research. Herein, utilizing multiple liquid chromatography-mass spectrometry (LC-MS)-based metabolomics approaches, we comprehensively characterized its components. We first systematized both LC and MS characteristics of polymethoxyflavones (PMFs), by which 13 PMFs were identified in C. aurantium, and their biosynthesis pathway was further established. Using derivatization-LC-MS targeted metabolomics approaches, 28 carbohydrates and 18 carboxylic acids were firstly found in C. aurantium. Combined with untargeted metabolomics method, total 147 compositions were characterized, among which 92 were firstly reported in C. aurantium. We further obtained their geographical features and sought out principal discriminative compounds. Moreover, typical biofunctions of C. aurantium from diverse regions were speculated using pharmacological platform and partly verified by experiments. The present study provided systematic component information for C. aurantium, which laid the foundation for its further exploitation as functional food.

Transcriptome and Metabolome Analysis Provides Insights into the Heterosis of Yield and Quality Traits in Two Hybrid Rice Varieties (Oryza sativa L.).

Heterosis is a common biological phenomenon that is useful for breeding superior lines. Using heterosis to increase the yield and quality of crops is one of the main achievements of modern agricultural science. In this study, we analysed the transcriptome and metabolome of two three-line hybrid rice varieties, Taiyou 871 (TY871), and Taiyou 398 (TY398) and the parental grain endosperm using RNA-seq (three biological repeats per variety) and untargeted metabolomic (six biological repeats per variety) methods. TY871 and TY398 showed specific heterosis in yield and quality. Transcriptome analysis of the hybrids revealed 638 to 4059 differentially expressed genes in the grain when compared to the parents. Metabolome analysis of the hybrids revealed 657 to 3714 differential grain metabolites when compared to the parents. The honeydew1 and grey60 module core genes Os04g0350700 and Os05g0154700 are involved in the regulation of awn development, grain size, and grain number, as well as the regulation of grain length and plant height, respectively. Rice grain length may be an important indicator for improving the quality of three-line hybrid rice. In addition, the rice quality-related metabolite NEG_M341T662 was highly connected to the module core genes Os06g0254300 and Os03g0168100. The functions of Os06g0254300 and Os03g0168100 are EF-hand calcium binding protein and late embroideries absolute protein repeat containing protein, respectively. These genes may play a role in the formation of rice quality. We constructed a gene and metabolite coexpression network, which provides a scientific basis for the utilization of heterosis in producing high-yield and high-quality hybrid rice.

Decreased IGF-1 level is associated with restrained amino acid metabolism in NSCLC with diabetes mellitus.

The discovery of a large number of small pulmonary nodules and early diagnosis of lung cancer in the diabetic patients prompt us to re-examine the relationship between diabetes and the occurrence and development of lung cancer. The aim of this study was to explore the underlying metabolites changes in diabetes with NSCLC or benign nodule patients, and further to investigate the association of serum IGF-1 level and differentially expressed metabolites (DEMs). An untargeted metabolomics method was used to detect the changes of metabolism in diabetic patients with NSCLC on the platform of HR-MS. Serum level of IGF-1 was measured by ELISA. The patients were divided to three groups, DM, DLB (nodule), and DLC (cancer). we have identified numerous DEMs, which include amino acid, choline, and fatty acid derivatives. Further analysis of the involved metabolic pathways suggested that linoleate metabolism, tryptophan metabolism, histidine metabolism, putative anti-Inflammatory metabolites formation from EPA, and arachidonic acid metabolism were considered to be the most significant metabolic pathways between groups. Networks analysis suggested that a series of metabolites were associated with serum IGF-1among the three groups, which can be divided into 6 categories. Nine metabolites have been identified as the main DEMs among the DLC, DLB, and DM groups. In conclusion, metabolomics is a powerful and promising tool for the cancer risk evaluation in diabetic patients. Our results suggest that decreased IGF-1 level is associated with restrained amino acid metabolism in NSCLC with diabetes mellitus.