Genome-Scale Analysis of the Structure and Function of RNA Pathways and Networks in Pseudomonas aeruginosa.

In recent years, several genome-wide approaches based on RNA sequencing (RNA-seq) have been developed. These methods allow a comprehensive and dynamic view of the structure and function of the multi-layered RNA pathways and networks. Many of these approaches, including the promising one of single-cell transcriptome analysis, have been successfully applied to Pseudomonas aeruginosa. However, we are only at the beginning because only a few surrounding conditions have been considered. Here, we aim to illustrate the different types of approaches based on RNA-seq that will lead us in the future to a better understanding of the dynamics of RNA biology in P. aeruginosa.

De Novo Transcriptomic Analyses to Identify and Compare Allergens in Foods.

Food allergens have been traditionally identified using biomolecular and immunological approaches. However, the techniques used in extracting proteins from the food source to be analyzed may hinder the availability of all proteins when assessing immunological allergenicity. Additionally, depending on the number and pool of patient sera used to detect the IgE antibody-binding allergens, some allergens may not be detected if not all the patients in the pool are sensitized to all the allergens. To overcome these limitations, we describe an additional approach before the in vitro approaches, by analyzing the transcriptome in silico for all putative allergens within the analyzed food source.

Combining transcriptomics and network pharmacology to reveal the mechanism of Zuojin capsule improving spasmolytic polypeptide-expressing metaplasia.

ETHNOPHARMACOLOGICAL RELEVANCE: Spasmolytic polypeptide-expressing metaplasia (SPEM) is a gastric precancerous lesion (GPL). Zuojin capsule (ZJC), consisting of Coptis chinensis Franch. (Ranunculaceae, recorded in the Chinese Pharmacopoeia as Rhizoma Coptidis) and Tetradium ruticarpum (A.Juss.) T.G.Hartley (Rutaceae, recorded in the Chinese Pharmacopoeia as Fructus Evodiae), has long been used for various gastrointestinal diseases. However, the effect and mechanism of ZJC on SPEM remain unclear. AIM OF THE STUDY: To clarify the role of ZJC in improving SPEM and study its mechanism. MATERIALS AND METHODS: The study utilized SPEM mice induced by 250 mg/kg body weight of tamoxifen (TAM) to assess the effects of ZJC and investigate its possible mechanisms. A strategy of transcriptomics combined with network pharmacology was conducted to explore the targets and mechanisms of ZJC in improving SPEM. The "ingredients-target-pathway" network was constructed, and the possible connections were verified by RT-qPCR and Western blot assays. RESULTS: ZJC significantly attenuated the abnormal serological indices, destruction of the gastric mucosal structure, hyperplasia of gastric pits, increased gastric mucus, massive secretion of CD44 and TFF2, oxyntic atrophy and massive proliferation of stem/progenitor cells in TAM-induced SPEM mice. Combined transcriptomics and network pharmacology analysis, 50 core targets of ZJC related to SPEM improvement were obtained. KEGG results showed that the core targets were significantly enriched in the cell cycle, and PI3K-AKT signaling pathway. The top-ranked targets according to PPI network analysis were CDK1, CCNB1, and CCNA2, which are also associated with cell cycle. Combined experiments demonstrated that ZJC can induce G2/M phase cycle arrest and inhibit TAM-induced malignant proliferation by regulating abnormal activation of cell cycle-related proteins such as CDK1, CCNB1, CCNA2 and PI3K-AKT signaling pathways. CONCLUSIONS: ZJC may improve TAM-induced SPEM by inhibiting abnormal activation of cell cycle-related proteins (CDK1, CCNB1, CCNA2) and PI3K-AKT signaling pathway. This finding supports the use of ZJC, a famous traditional Chinese medicine compound, as a potential treatment for gastric precancerous lesions.

Analysis of Xylem Cells by Nucleus-Based Transcriptomics and Chromatin Profiling.

Nuclei contain essential information for cell states, including chromatin and RNA profiles - features which are nowadays accessible using high-throughput sequencing applications. Here, we describe analytical pipelines including nucleus isolation from differentiated xylem tissues by fluorescence-activated nucleus sorting (FANS), as well as subsequent SMART-seq2-based transcriptome profiling and assay for transposase-accessible chromatin (ATAC)-seq-based chromatin analysis. Combined with tissue-specific expression of nuclear fluorescent reporters, these pipelines allow obtaining tissue-specific data on gene expression and on chromatin structure and are applicable for a large spectrum of cell types, tissues, and organs. Considering, however, the extreme degree of differentiation found in xylem cells with programmed cell death happening during vessel element formation and their role as a long-term depository for atmospheric CO2 in the form of wood, xylem cells represent intriguing and relevant objects for large-scale profilings of their cellular signatures.

Comprehensive analysis of aflatoxin B1 biosynthesis in Aspergillus flavus via transcriptome-wide m6A methylome response to cycloleucine.

Aspergillus flavus and its toxic aflatoxins secondary metabolites contaminate food and grains, posing a severe threat to human health and leading to liver cancer. Here, we demonstrated that cycloleucine blocked aflatoxin B1 synthesis by inhibiting N6-methyladenosine (m6A) methylation modification of messenger RNA (mRNA). m6A Methylation Immunoprecipitation Sequencing (m6A MeRIP-Seq)-based comprehensive transcriptome-wide m6A profiling identified 102 differentially expressed genes that underwent m6A modification, of which 22 hypermethylated genes were downregulated and 49 hypomethylated genes were upregulated, suggesting a negative correlation between m6A methylation and gene expression. Notably, cycloleucine inhibited aflatoxin B1 production via multiple targets. The m6A sites of several key genes involved in the aflatoxin B1 biosynthesis pathway were significantly enriched in the coding sequence and around the stop codon, resulting in their downregulation. Furthermore, m6A methylation on genes related to the aflatoxin B1 biosynthesis pathway led to reduced mRNA stability. Cycloleucine inhibition of aflatoxin B1 production highlights its potential as an agent for removing mycotoxins in environmental pollution. ENVIRONMENTAL IMPLICATION: Aflatoxins, highly carcinogenic secondary metabolites produced by Aspergillus flavus, frequently contaminate crops such as peanut, corn, wheat and sesame leading to irreversible loss in the quality and yield of agricultural products and posing serious threats to food safety. Aflatoxins has also been linked to developmental delays and liver cancer in humans. In our study, 'monitoring aflatoxin concentrations and its bioaccumulation in organisms' has been conducted. The results demonstrated that aflatoxin production in A. flavus was completely blocked after cycloleucine treatment. Additionally, we demonstrated that inhibition of aflatoxin was linked to N6-methyladenosine methylation of multiple genes in aflatoxin biosynthesis pathway.

Network pharmacology and transcriptomics to determine Danggui Yifei Decoction mechanism of action for the treatment of chronic lung injury.

ETHNOPHARMACOLOGICAL RELEVANCE: Several children with pneumonia (especially severe cases) have symptoms of cough and expectoration during the recovery stage after standard symptomatic treatment, which eventually results in chronic lung injury. Danggui yifei Decoction (DGYFD), a traditional Chinese formula, has shown clinical promise for the treatment of chronic lung injury during the recovery stage of pneumonia, however, its mechanism of action is yet to be deciphered. AIM OF THIS STUDY: To investigate the therapeutic mechanism of DGYFD for the treatment of chronic lung injury by integrating network pharmacology and transcriptomics. MATERIALS AND METHODS: BALB/c mice were used to establish the chronic lung injury mouse model by intratracheal instillation of lipopolysaccharide (LPS). Pathological analysis of lung tissue, lung injury histological score, lung index, protein levels in bronchoalveolar lavage fluid (BALF), immunohistochemical staining, blood rheology, inflammatory cytokines, and oxidative stress levels were used to evaluate the pharmacological effects of DGYFD. Chemical components of DGYFD were identified using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Integrated network pharmacology together with transcriptomics was used to predict potential biological targets. Western blot analysis was used to verify the results. RESULTS: In this study, we demonstrated that DGYFD could improve lung injury pathological changes, decreases lung index, down-regulate NO and IL-6 levels, and regulate blood rheology. In addition, DGYFD was able to reduce the protein levels in BALF, up-regulate the expression levels of occludin and ZO-1, improve the ultrastructure of lung tissues, and reverse the imbalance of AT I and AT II cells to repair the alveolar-capillary permeability barrier. Twenty-nine active ingredients of DGYFD and 389 potential targets were identified by UPLC-MS/MS and network pharmacology, and 64 differentially expressed genes (DEGs) were identified using transcriptomics. GO and KEGG analysis revealed that the MAPK pathway may be the molecular target. Further, we found that DGYFD inhibits phosphorylation levels of p38 MAPK and JNK in chronic lung injury mouse models. CONCLUSIONS: DGYFD could regulate the imbalance between the excessive release of inflammatory cytokines and oxidative stress, repair the alveolar-capillary permeability barrier and improve the pathological changes during chronic lung injury by regulating the MAPK signaling pathway.

Mechanism of Xiaojianzhong decoction in alleviating aspirin-induced gastric mucosal injury revealed by transcriptomics and metabolomics.

ETHNOPHARMACOLOGICAL RELEVANCE: Aspirin, as a first-line drug for the treatment of cardiovascular diseases, currently has high clinical usage. However, reports of aspirin-induced gastric mucosal injury are increasing. Xiaojianzhong decoction (XJZD), a classic traditional Chinese medicine formula, has been shown to alleviate gastric mucosal injury, although its potential mechanism of action requires further study. AIM OF THE STUDY: This study aimed to explore the effect and mechanism of XJZD in preventing aspirin-induced gastric mucosal injury. MATERIALS AND METHODS: Aspirin was used to induce damage in the morning, while XJZD was applied as an intervention in the afternoon. The compounds in the XJZD were analyzed by means of both high-performance liquid chromatography and ultra-performance liquid chromatography-tandem mass spectrometry. The overall condition of the aspirin-related gastric mucosal injury was evaluated. The expressions of inflammatory factors and tight-junction-related proteins and apoptosis were observed via immunohistochemistry and immunofluorescence. The expression levels of the apoptosis-related proteins were detected using Western blot. Transcriptomics was used to perform the integrative analysis of gastric tissues, which was then validated. Molecular dynamics was used to explore the interaction of key compounds within the XJZD with relevant targets. Finally, non-targeted metabolomics was used to observe any metabolic changes and construct a network between the differentially expressed genes and the differential metabolites to elucidate their potential relationship. RESULTS: XJZD can alleviate inflammation response, maintain the gastric mucosal barrier's integrity, reduce apoptosis and necroptosis levels, and promote the proliferation and repair of gastric mucosal tissues. Its mechanism of action may be related to the regulation of TNF-α signaling. Furthermore, molecular docking showed that the cinnamaldehyde within XJZD played an important role in its effects. In addition, XJZD can correct metabolic disorders, mainly regulating amino acid metabolism pathways. Moreover, six differential genes (Cyp1a2, Cyp1a1, Pla2g4c, etc.) were determined to alleviate both gastric mucosal injury and inflammation by regulating arachidonic acid metabolism, Tryptophan metabolism, etc. CONCLUSIONS: This study is the first to report that XJZD can inhibit necroptosis and gastric mucosal injury induced by aspirin, thereby revealing the complex mechanism of XJZD in relation to alleviating gastric mucosal injury from multiple levels and perspectives.

Physiological and transcriptomic insights into sugar stress resistance in osmophilic yeast Zygosaccharomyces rouxii.

The osmophilic yeast Zygosaccharomyces rouxii has attracted increasing attention for its ability to survive and grow in extremely high sugar environments. This trait determines its role in fermentation process and results in contamination in the food industry. However, the behavior of Z. rouxii in regulating cell metabolism to combat high sugar stress and the corresponding mechanism have not been completely elucidated. Here, the resistance strategies of Z. rouxii against high glucose stress were explored by physiological analysis at cell membrane level and transcriptomic analysis. Physiological analysis showed that under high glucose stress, colony transparency increased, cell volume decreased, which was accompanied by reduction in permeability and integrity of cell membrane and subsequent gradual recovering. Additionally, the proportion of ergosterol and unsaturated fatty acids in cell membrane significantly increased under high glucose stress. A comparison of transcriptome data showed that most of the obtained differentially expressed genes (DEGs) involved in ergosterol and linoleic acid synthesis pathways as well as cell wall integrity (CWI) and high osmolarity glycerol mitogen-activated protein kinase (HOG-MAPK) pathways, which was in line with the results of physiological data. Our results provided a theoretic basis to develop the process control for the production of high sugar foods.

Insights into the reduction of methylmercury accumulation in rice grains through biochar application: Hg transformation, isotope fractionation, and transcriptomic analysis.

Methylmercury (MeHg), a potent neurotoxin, easily moves from the soil into rice plants and subsequently accumulates within the grains. Although biochar can reduce MeHg accumulation in rice grains, the precise mechanism underlying biochar-mediated responses to mercury (Hg) stress, specifically regarding MeHg accumulation in rice, remains poorly understood. In the current study, we employed a 4% biochar amendment to remediate Hg-contaminated paddy soil, elucidate the impacts of biochar on MeHg accumulation through a comprehensive analysis involving Hg isotopic fractionation and transcriptomic analyses. The results demonstrated that biochar effectively lowered the levels of MeHg in paddy soils by decreasing bioavailable Hg and microbial Hg methylation. Furthermore, biochar reduced the uptake and translocation of MeHg in rice plants, ultimately leading to a reduction MeHg accumulation in rice grains. During the process of total mercury (THg) uptake, biochar induced a more pronounced negative isotope fractionation magnitude, whereas the effect was less pronounced during the upward transport of THg. Conversely, biochar caused a more pronounced positive isotope fractionation magnitude during the upward transport of MeHg. Transcriptomics analyses revealed that biochar altered the expression levels of genes associated with the metabolism of cysteine, glutathione, and metallothionein, cell wall biogenesis, and transport, which possibly enhance the sequestration of MeHg in rice roots. These findings provide novel insights into the effects of biochar application on Hg transformation and transport, highlighting its role in mitigating MeHg accumulation in rice.

Transcriptome and ultrastructural analysis revealed the mechanism of Mercapto-palygorskite on reducing Cd content in wheat.

Large areas of crop yields in northern China have faced with cadmium (Cd) contamination problems. Mercapto-modified palygorskite (MP), as a highly efficient immobilization material, could reduce Cd absorption in wheat and alleviate its biotoxicity. However, the molecular mechanism underlying MP-mediated Cd reduction and detoxification processes in wheat is not well understood. This aim of this study was to investigate the biochemical and molecular mechanisms underlying the reduction in Cd accumulation in wheat (Triticum aestivum L.). The results showed that MP application decreased the Cd concentration by 68.91-74.32% (root) and 70.68-77.2% (shoot), and significantly increased the glutathione (GSH) and phytochelatins (PCs) contents in root and shoot. In addition, with the application of MP, the percentage of Cd in the cell walls and organelles of wheat decreased, while that of Cd in soluble components was increased. The content of Cd in all components was significantly reduced. Ultrastructural analysis revealed that MP thickened the cell wall, promoted vesicle formation in the membrane and protected the integrity of intracellular organelles in wheat. Transcriptome analysis further confirmed the above results. MP upregulated the expression of several genes (CCR, CAD COMT and SUS) involved in cell wall component biosynthesis and promoted vesicle formation on cell membranes by upregulating the expression of PLC and IPMK genes. In addition, genes related to antioxidant synthesis (PGD, glnA and GSS) and photosynthesis (Lhca, Lhcb) were altered by MP to alleviate Cd toxicity in wheat. This present work will help to more thoroughly elucidate the molecular mechanism by which wheat defends against Cd contamination under MP application and provide and important research basis for the application of this material in the future.

Single-cell transcriptomics reveals the pulmonary inflammation induced by inhalation of subway fine particles.

People generally take the subway and inevitably inhale the fine particles (PM2.5) on subway platforms. This study revealed whether and how subway PM2.5 causes lung inflammation. Herein, the pulmonary inflammatory response to subway PM2.5 was observed in mice, manifesting as the inflammatory cells infiltration and collagen deposition in tissue, inflammatory cytokine enhancement in bronchoalveolar lavage fluid and Toll-like receptors signal pathway activation in the lungs. Furthermore, single-cell RNA sequencing unearthed subway PM2.5-induced cell-specific responses in the lungs. Twenty immune subsets were identified by the molecular and functional properties. Specific cell populations of CD4+ T and γδ T cells were regarded as the predominant sources of pneumonitis induced by subway PM2.5. Moreover, we demonstrated that the lung inflammatory injury was significantly more attenuated in Rag1-/- mice lacking functional T cells and B cells than that in wild type mice. We proved the slight inflammation of lung tissue in Rag1-/- mice may be dependent on monocytes and neutrophils by activation of the intracellular molecular network. This is the first experimental study on subway PM2.5 causing pulmonary inflammatory damage. It will set an alarm for people who usually travel by subway and efficient measures to reduce PM2.5 should be developed in subway stations.

A novel strategy of integrating network pharmacology and transcriptome reveals antiapoptotic mechanisms of Buyang Huanwu Decoction in treating intracerebral hemorrhage.

ETHNOPHARMACOLOGICAL RELEVANCE: Buyang Huanwu Decoction (BYHWD), as a traditional Chinese medical prescription, has been used to treat intracerebral hemorrhage (ICH) for hundreds of years, but the antiapoptotic properties have not yet been studied. AIM OF THE STUDY: This study aims to elucidate the antiapoptotic mechanism of BYHWD in ICH. MATERIALS AND METHODS: The therapeutic effect of BYHWD on ICH was assessed by modified neurological severity scores (mNSS), foot fault, and histopathological staining. Then, we used a modified comprehensive strategy by integrating transcriptome and network pharmacology to reveal the underlying mechanism. TUNEL assay, qRT-PCR, and western blot were further applied to evaluate the antiapoptotic effect of BYHWD on ICH. Dual-luciferase reporter assay and plasmid transfections were implemented to validate the potential competing endogenous RNAs (ceRNA) mechanism of Sh2b3. RESULTS: Network pharmacology analysis indicated that the regulation of the apoptotic process was the highest enriched GO term, and that MAP kinase activity, ERK1, and ERK2 cascade were strongly correlated. Transcriptome analysis screened 180 differentially expressed mRNAs, which were highly enriched in the immune system process and negative regulation of programmed cell death. By checking the literature, we found that Sh2b3 was of great importance to apoptosis by modulating MAPK cascades. TUNEL assay validated the anti-apoptotic effect of BYHWD. Moreover, BYHWD was proven to regulate the Sh2b3-mediated ERK1/2 signaling pathway in ICH mice by qRT-PCR and western blot. We further explored the lncRNA-miRNA-mRNA network underlying the therapeutic effect, among which 4933404O12Rik/miR-185-5p is the upstream regulatory mechanism of Sh2b3. CONCLUSIONS: We explored the antiapoptotic mechanism of BYHWD in treating ICH by a novel integrated strategy, which involved the 4933404O12Rik/miR-185-5p/Sh2b3 ceRNAs axis.

Lineage Tracing by Single-Cell Transcriptomics Decoding Dynamics of Lineage Commitment.

Tracing the fate of individual cells and their progeny is necessary and significant for stem cell research and cancer research. Changes in lineage-specific transcription factor levels during lineage commitment are gradual and continuous. Development of single-cell sequencing technology allows many different states of cells to be sequenced at an unprecedented resolution, and it has been proved that single-cell transcriptomics meets lineage tracing. Here, we introduce a detailed protocol for the lineage tracing by single-cell transcriptomics to clarify the dynamics of lineage commitment.

Metastasis and recurrence patterns in the molecular subtypes of urothelial bladder cancer.

Urothelial cancer of the urinary bladder frequently metastasizes to lymph-nodes, lungs, liver and bone. A taxonomy for molecular classification exists, but it is unknown if molecular subtypes show tropism for different organs. Here, we study 146 patients with de novo metastatic disease or recurrence after curative treatment. We classify primary tumors using two transcriptomic methods and immunostaining and identify enrichment and depletion of metastatic sites in molecular subtypes using permutation tests. We observed significant depletion of bone metastases in the Basal/squamous molecular subtype, whereas the Urothelial-like subtype entailed an enrichment for metastases to bone. The Genomically unstable subtype was depleted of lung metastases, but enriched for atypical sites, including six out of seven patients with brain metastases. Stroma-rich primary tumor samples were associated with local recurrence, but not with distant sites. Additionally, the proportion with brain or testis metastases differed between systemic chemotherapy regimens (GC vs MVAC) suggesting a sanctuary effect. In conclusion, molecular subtypes of urothelial bladder cancer are significantly associated with specific metastatic sites, suggesting that subtype-specific molecular determinants could exist at various steps in the metastatic cascade.

Using regular and transcriptomic analyses to investigate the biotransformation mechanism and phytotoxic effects of 6:2 fluorotelomer carboxylic acid (6:2 FTCA) in pumpkin (Cucurbita maxima L.).

Although 6:2 fluorotelomer carboxylic acid (6:2 FTCA), which is one of the most popular substitutes for perfluorooctanoic acid (PFOA), has been widely distributed in environments, little is known about its biotransformation mechanism and phytotoxic effects in plants. Here, we showed that 6:2 FTCA could be taken up by pumpkin (Cucurbita maxima L.) roots from exposure solution and acropetally translocated to shoots. Biotransformation of 6:2 FTCA to different carbon chain perfluorocarboxylic acid (PFCA) metabolites (C2-C7) via α-and ß-oxidation in pumpkin was observed, and perfluorohexanoic acid (PFHxA) was the major transformation product. The results of enzyme assays, enzyme inhibition experiments and gene expression analysis indicated that cytochrome P450 (CYP450), glutathione-S-transferase (GST) and ATP-binding cassette (ABC) transporters were involved in the metabolism of 6:2 FTCA in pumpkin. Plant-associated rhizobacteria and endophyte also contributed to 6:2 FTCA degradation through ß-oxidation. The chlorophyll (Chl) content and genes involved in photosynthesis were significantly improved by 6:2 FTCA. The reductions of antioxidant and metabolic enzyme activities reflected the antioxidant defense system and detoxification system of pumpkin were both damaged, which were further confirmed by the down-regulating associated genes encoding phenylpropanoid biosynthesis, endoplasmic reticulum-related proteins, ascorbate-glutathione cycle and ABC transporters. This study is helpful to understand the environmental behaviors and toxicological molecular mechanisms of 6:2 FTCA in plants.

Physiological, transcriptome and gene functional analysis provide novel sights into cadmium accumulation and tolerance mechanisms in kenaf.

Kenaf is considered to have great potential for remediation of heavy metals in ecosystems. However, studies on molecular mechanisms of root Cd accumulation and tolerance are still inadequate. In this study, two differently tolerant kenaf cultivars were selected as materials and the physiological and transcriptomic effects were evaluated under Cd stress. This study showed that 200 µmol/L CdCl2 treatment triggered the reactive oxygen species (ROS) explosion and membrane lipid peroxidation. Compared with the Cd-sensitive cultivar 'Z', the Cd-tolerant cultivar 'F' was able to resist oxidative stress in cells by producing higher antioxidant enzyme activities and increasing the contents of ascorbic acid (AsA) and glutathione (GSH). The root cell wall of 'F' exhibited higher polysaccharide contents under Cd treatment, providing more Cd-binding sites. There were 3,439 differentially expressed genes (DEGs) that were co-regulated by Cd treatment in two cultivars. Phenylpropanoid biosynthesis and plant hormone signal transduction pathways were significantly enriched by functional annotation analysis. DEGs associated with pectin, cellulose, and hemi-cellulose metabolism were involved in Cd chelation of root cell wall; V-ATPases, ABCC3 and Narmp3 could participated in vacuolar compartmentalization of Cd; PDR1 was responsible for Cd efflux; the organic acid transporters contributed to the absorption of Cd in soil. These genes might have played key roles in kenaf Cd tolerance and Cd accumulation. Moreover, HcZIP2 was identified to be involved in Cd uptake and transport in kenaf. Our findings provide a deeper understanding of the molecular pathways underlying Cd accumulation and detoxification mechanisms in kenaf.

MICA: a multi-omics method to predict gene regulatory networks in early human embryos.

Recent advances in single-cell omics have transformed characterisation of cell types in challenging-to-study biological contexts. In contexts with limited single-cell samples, such as the early human embryo inference of transcription factor-gene regulatory network (GRN) interactions is especially difficult. Here, we assessed application of different linear or non-linear GRN predictions to single-cell simulated and human embryo transcriptome datasets. We also compared how expression normalisation impacts on GRN predictions, finding that transcripts per million reads outperformed alternative methods. GRN inferences were more reproducible using a non-linear method based on mutual information (MI) applied to single-cell transcriptome datasets refined with chromatin accessibility (CA) (called MICA), compared with alternative network prediction methods tested. MICA captures complex non-monotonic dependencies and feedback loops. Using MICA, we generated the first GRN inferences in early human development. MICA predicted co-localisation of the AP-1 transcription factor subunit proto-oncogene JUND and the TFAP2C transcription factor AP-2γ in early human embryos. Overall, our comparative analysis of GRN prediction methods defines a pipeline that can be applied to single-cell multi-omics datasets in especially challenging contexts to infer interactions between transcription factor expression and target gene regulation.

Integrated analysis of metabolomic and transcriptomic profiling reveals the effect of Atractylodes oil on Spleen Yang Deficiency Syndrome in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Spleen Yang Deficiency Syndrome (SYDS), which is a syndrome commonly treated with Traditional Chinese Medicine (TCM), manifests as overall metabolic dysfunction caused mainly by digestive system disorders. Atractylodes lancea (Thunb.) DC. (AL) is a widely used traditional herb with the efficacy of eliminate dampness and strengthen the spleen, Atractylodes oil (AO) is a medicinal component of AL and can be used to treat various gastrointestinal disorders. However, its effects on SYDS and underlying mechanisms have not been clarified to date. AIM OF THE STUDY: The present study aimed to investigate the efficacy of AO in the improvement of the symptoms of SYDS in rat and the underlying mechanism by integrating transcriptomics, and metabolomics. MATERIALS AND METHODS: The SYDS rats induced by reserpine were treated with AO. The protective effect of AO on SYDS rats was evaluated by serum biochemical detection, histopathological analyses. Enzyme-linked immunosorbent assay (ELISA), colorimetric assay and immunofluorescence (IF) were performed to determine the levels of relevant indicators of mitochondrial function and energy metabolism in the liver. Liver metabolites and transcript levels were assessed by non-targeted metabolomics and transcriptomics to analyze potential molecular mechanisms and targets. The expression of the corresponding proteins was verified using Western blotting. RESULTS: AO not only regulated the digestion, absorption function and oxidative stress status of SYDS rats, but also improved mitochondrial function and alleviated energy metabolism disorders in SYDS rats. Metabolomic and transcriptomic analyses demonstrated that AO regulation is mainly exerted in amino acid metabolism, unsaturated fatty acid metabolism, TCA cycle as well as PPAR and AMPK signaling pathways. In addition, The AMPK signaling pathway was verified and AO promoted AMPK phosphorylation and the expression of SIRT1, PGC-1α, and PPARα in SYDS rats. CONCLUSIONS: The therapeutic effect of AO on SYDS is potentially attributable to activation of the AMPK/SIRT1/PGC-1α signaling pathway, which enhances transport and regulation of energy metabolism.

Transcriptome wide identification of neuropeptides and G protein-coupled receptors (GPCRs) in Sunn pest, Eurygaster integriceps Puton.

Sunn pest (Eurygaster integriceps Puton) is major wheat pest causing economic damage. Neuropeptides and their receptors, G protein-coupled receptors (GPCRs), are involved in the regulation of insect physiology and behavior. Herein, a transcriptome-wide analysis was conducted in order to identify genes encoding neuropeptides, and putative GPCRs to gain insight into neuropeptide-modulated processes. De novo transcriptome assembly was undertaken using paired-end sequence reads derived from RNA samples collected from whole adults and yielded 582,398 contigs. In total, 46 neuropeptides have been identified, encompassing various known insect neuropeptide families. In addition, we discovered four previously uncharacterized neuroparsin peptides, which contributes to our understanding of the neuropeptide landscape. Furthermore, 85 putative neuropeptide GPCRs were identified, comprising three classes of GPCRs, A, B, C, and LGR, of which class C is not widely reported in insects. In addition, the identified GPCRs exhibited a remarkable 80% homology with the GPCRs found in the brown marmorated stink bug. It is noteworthy that these GPCRs displayed only a 20% homology to GPCRs from many other insect species. This information may be used to understand the neuropeptide-modulated physiology and behavior of Eurygaster integriceps, and to develop specific neuropeptide-based pest management strategies.

Bark transcriptome analyses reveals molecular mechanisms involved in tapping panel dryness occurrence and development in rubber tree (Hevea brasiliensis).

Tapping panel dryness (TPD) has become the mostimportant limiting factor for increasing natural rubber yield, whereas illuminating the molecular mechanisms underlying TPD is the prerequisite for solving the problem of TPD. However, molecular mechanisms underlying TPD are largely unknown. In this study, healthy and different stages of TPD-affected rubber trees were utilized to analyze TPD for the first time. We found that the changing tendencies of key latex physiological parameters were closely related to TPD occurrence and development. To reveal the molecular mechanisms underlying TPD, we sequenced and compared bark transcriptomes among healthy rubber tree, and TPD-affected ones at initial and advanced stages. In total, 8607 genes were identified as TPD-related genes in contrast to healthy rubber tree. According to gene expression profiles, the five samples were divided into three groups including healthy rubber tree, and TPD-affected rubber tree in the initial and advanced stages, which was consistent with the stages of TPD occurrence and development. Interestingly, only asmall proportionof the TPD-related genes were constantly down- or up-regulated with TPD occurrence and development. The TPD-related genes in KEGG pathways significantly enriched were closely associated with protein metabolism, cell division and differentiation, PCD, stress responses, terpene biosynthesis, and various metabolism processes. Moreover, overexpression of HbAPX2 identified as a TPD-related gene enhanced oxidative stress tolerance in S. cerevisiae. The typical symptoms of TPD, partial or complete dry zone (no latex flow) on tapping panel, might attribute to lower IPP available for rubber biosynthesis, and downregulation of the genes in post-IPP steps of rubber biosynthesis and the genes involved in latex flow. Our results not only provide new insights into molecular mechanisms underlying TPD occurrence and development but also contribute to developing effective measures to control TPD in rubber trees.