The high-quality sequencing of the Brassica rapa 'XiangQingCai' genome and exploration of genome evolution and genes related to volatile aroma.

'Vanilla' (XQC, brassica variety chinensis) is an important vegetable crop in the Brassica family, named for its strong volatile fragrance. In this study, we report the high-quality chromosome-level genome sequence of XQC. The assembled genome length was determined as 466.11 Mb, with an N50 scaffold of 46.20 Mb. A total of 59.50% repetitive sequences were detected in the XQC genome, including 47 570 genes. Among all examined Brassicaceae species, XQC had the closest relationship with B. rapa QGC ('QingGengCai') and B. rapa Pakchoi. Two whole-genome duplication (WGD) events and one recent whole-genome triplication (WGT) event occurred in the XQC genome in addition to an ancient WGT event. The recent WGT was observed to occur during 21.59-24.40 Mya (after evolution rate corrections). Our findings indicate that XQC experienced gene losses and chromosome rearrangements during the genome evolution of XQC. The results of the integrated genomic and transcriptomic analyses revealed critical genes involved in the terpenoid biosynthesis pathway and terpene synthase (TPS) family genes. In summary, we determined a chromosome-level genome of B. rapa XQC and identified the key candidate genes involved in volatile fragrance synthesis. This work can act as a basis for the comparative and functional genomic analysis and molecular breeding of B. rapa in the future.

Integrated fecal microbiota and metabolomics analysis of the orlistat intervention effect on polycystic ovary syndrome rats induced by letrozole combined with a high-fat diet.

BACKGROUND: This study aimed to compare the characteristics of the gut microbiota and their metabolite profiles between polycystic ovary syndrome (PCOS) and orlistat-treated PCOS rats (ORL-PCOS), which could help to better understand the underlying mechanism of the effect of orlistat on PCOS. METHODS: PCOS rat models were established using letrozole combined with a high-fat diet. Ten rats were randomly selected as a PCOS control group (PCOS). The other three groups (n = 10/group) were additionally supplemented with different doses of orlistat (low, medium, high). Then, fecal samples of the PCOS and ORL-PCOS groups were analysed by 16S rRNA gene sequencing and untargeted metabolomics. Blood samples were collected to detect serum sex hormones and lipids. RESULTS: The results showed that orlistat attenuated the body weight gain, decreased the levels of T, LH, the LH/FSH ratio, TC, TG and LDL-C; increased the level of E2; and improved estrous cycle disorder in PCOS rats. The bacterial richness and diversity of the gut microbiota in the ORL-PCOS group were higher than those in the PCOS group. The ratio of Firmicutes to Bacteroidetes was decreased with orlistat treatment. Moreover, orlistat treatment led to a significant decrease in the relative abundance of Ruminococcaceae and Lactobacillaceae, and increases in the abundances of Muribaculaceae and Bacteroidaceae. Metabolic analysis identified 216 differential fecal metabolites in total and 6 enriched KEGG pathways between the two groups, including steroid hormone biosynthesis, neuroactive ligand-receptor interaction and vitamin digestion and absorption. Steroid hormone biosynthesis was the pathway with the most significant enrichment. The correlations between the gut microbiota and differential metabolites were calculated, which may provide a basis for understanding the composition and function of microbial communities. CONCLUSIONS: Our data suggested that orlistat exerts a PCOS treatment effect, which may be mediated by modifying the structure and composition of the gut microbiota, as well as the metabolite profiles of PCOS rats.

Echinacea purpurea-derived homogeneous polysaccharide exerts anti-tumor efficacy via facilitating M1 macrophage polarization.

Echinacea purpurea modulates tumor progression, but the underlying mechanism is poorly defined. We isolated and purified a novel homogeneous polysaccharide from E. purpurea (EPPA), which was shown to be an arabinogalactan with a mean molecular mass (Mr) of 3.8 × 104 Da and with α- (1 → 5) -L-Arabinan as the backbone and α-L-Araf-(1→, →6)-ß-D-Galp-(1→, and →4)-α-D-GalpA-(1→ as the side chains. Interestingly, oral administration of EPPA suppresses tumor progression in vivo and shapes the immune cell profile (e.g., facilitating M1 macrophages) in tumor microenvironment by single-cell RNA sequencing (scRNA-seq) analysis. More importantly, EPPA activates the inflammasome through a phagocytosis-dependent mechanism and rewires transcriptomic and metabolic profile, thereby potentiating M1 macrophage polarization. Collectively, we propose that EPPA supplementation could function as an adjuvant therapeutic strategy for tumor suppression.

Assessment of Significant Pathway Signaling and Prognostic Value of GNG11 in Ovarian Serous Cystadenocarcinoma.

BACKGROUND: GNG11 (G protein subunit gamma 11) is a member of guanine nucleotide-binding protein (G protein) gamma family. Few studies elucidated the role of GNG11 in human disease, especially in tumors. The present study initially analyzed the function of GNG11 in ovarian serous cystadenocarcinoma. METHODS: The differential expression of GNG11 mRNA in ovarian cancer and normal tissues was evaluated through Oncomine, CCLE, Gepia, UCSC Xena and UALCAN databases. The protein expression of GNG11 was assessed via HPA database. Prognosis analysis was performed by Kaplan-Meier Plotter. Restrict survival analysis to subtypes including tumor grade, cancer stage and TP53 mutation status was then carried out. GSEA enrichment analysis was performed to explore the significant pathways associated with GNG11 in ovarian cancer. Finally, the upstream miRNAs of GNG11 were predicted by DIANA, Target Scan, miRDB and miRWalk databases, and the potential key KEGG pathways were subsequently determined by DIANA. RESULTS: The mRNA expression of GNG11 was down-regulated in ovarian cancer patients (P<0.05). The cancer stage of patients correlated with the expression of GNG11 (P<0.05). Survival analysis indicated that GNG11 high expression statistically shortened the overall survival time of patients (HR=1.26, P=0.0043) compared with low expression group, especially for the patients with earlier stage (HR=2.48, P=0.035) and lower grade (HR=1.72, P=0.0016). Subsequently, the consistent upstream miRNA of GNG11, hsa-miR-22-5p, was predicted from 4 databases. The differential expression profile of hsa-miR-22-5p in blood was observed in ovarian cancer patients. According to the GSEA analysis on GNG11 and KEGG analysis on hsa-miR-22-5p, the consistent pathway of ECM-receptor interaction was observed (all P<0.01). ECM-receptor interaction pathway and differential expression of hsa-miR-22-5p in blood suggested the migration risk of ovarian cancer. CONCLUSION: High expression of GNG11 indicated the poor prognosis of ovarian cancer patients. GNG11 might play a crucial role in the biological process of ovarian serous cystadenocarcinoma by ECM-receptor interaction pathway, thus affecting the prognosis of patients.

Phillygenin exhibits anti-inflammatory activity through modulating multiple cellular behaviors of mouse lymphocytes.

Context: Phillygenin (PHI) is an intestinal metabolite of phillyrin from the genus Forsythia. Although the regulatory activity of Forsythia on immune system has been investigated, the effect of PHI on activated lymphocytes is poorly understood. Objective: This study was aimed to discuss the possible anti-inflammation potential of PHI on mitogen-activated stimulated lymphocytes in vitro. Methods: Mice spleen lymphocytes were incubated with PHI for 4 h, and then stimulated with concanavalin A (Con A) or phorbol 12-myristate 13-acetate/ionomycin (PMA + Ion). Cell viability was assayed by cell counting kit-8 (CCK-8). The expression of CD69 and CD25, proliferation, cell cycle, intracellular Ca2+ concentration, apoptosis, mitochondrial inner membrane potential (ΔΨm), mitochondrial permeability transition (MPT), interleukin-2 (IL-2), interferon-γ (IFN-γ), and tumor necrosis factor-α (TNF-α) were analyzed by flow cytometry. The expression of cyclin B1, cyclin D1, Cyclin E, and the phosphorylation of c-jun N-terminal kinase (JNK), extracellular signal-regulated kinase 1/2 (Erk1/2) and p38 were assayed by western blotting. Results: The results showed that PHI inhibited the proliferation of Con A-activated lymphocytes and induced a G0/G1 phase arrest by suppressing cyclin D1 and cyclin E. Meanwhile, PHI antagonized Con A-induced T cells activation through blocking intracellular Ca2+ overload and suppressing the phosphorylation of JNK and Erk1/2. Both Con A and PMA + Ion-induced secretion of IL-2, IFN-γ, and TNF-α were attenuated by PHI. PHI enhanced Con A-induced lymphocytes apoptosis through decreasing ΔΨm and increasing MPT. Conclusion: These results suggest that PHI exhibits its anti-inflammatory activity through modulating multiple cellular behaviors, leading to the suppression of the adaptive immune response.

Correction to: Transcriptome profiling of genes related to light-induced anthocyanin biosynthesis in eggplant (Solanum melongena L.) before purple color becomes evident.

After publication of the original article [1] it was noted that in Additional file 1: Table S1, and Fig. 1, specific primer sequences were incorrect, and taken from Sme2.5_02154.1_g00001.1 rather than Sme2.5_13923.1_g00001.1.

Transcriptome profiling of genes related to light-induced anthocyanin biosynthesis in eggplant (Solanum melongena L.) before purple color becomes evident.

BACKGROUND: The anthocyanins are highly enriched in eggplants (Solanum melongena L.) with purple peel. However, our previous study showed that anthocyanins biosynthesis in eggplant cultivar 'Lanshan Hexian' was completely regulated by light and color becomes evident at most 2 days after exposure to light. In the present investigation, transcriptome study was made to explore the underlying molecular mechanisms of light-induced anthocyanin biosynthesis in eggplant (Solanum melongena L.) before color becomes evident. RESULTS: RNA-Seq was performed for four time points (0, 0.5, 4 and 8 h after bags removal) where concerted changes happened. A total of 32,630 genes or transcripts were obtained by transcriptome sequencing, from which 1956 differentially expressed genes (DEGs) were found. Gene Ontology analysis showed that the 1956 DEGs covered a wide range of cellular components, molecular functions and biological processes. All the DEGs were further divided into 26 clusters based on their distinct expression patterns. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis found out 24 structural anthocyanin biosynthesis genes which distributing in seven clusters. In addition, 102 transcription factors, which exhibited highly dynamic changes in response to light, were found in the seven clusters. Three photoreceptors, UV Resistance Locus 8 (UVR8), Cryptochrome 3 (CRY3) and UVR3, were identified as DEGs. The light signal transduction elements, COP1 and two SPAs, might be responsible for anthocyanin biosynthesis regulation. CONCLUSION: Based on the transcriptome data, the anthocyanin biosynthesis structural genes, transcription factors, photoreceptors and light signal transduction elements were quickly screened which may act as the key regulatory factors in anthocyanin biosynthesis pathway. By comparing the transcriptome data with our previous studies, 869 genes were confirmed to participate in the light-induced anthocyanin biosynthesis. These results expand our knowledge of light-induced anthocyanin biosynthesis in plants, which allowing for fruit coloration to be improved under low-light conditions in future.

Combined transcriptomic and proteomic analysis constructs a new model for light-induced anthocyanin biosynthesis in eggplant (Solanum melongena L.).

Light is a key environmental factor affecting anthocyanin biosynthesis. Our previous study demonstrated that "Lanshan Hexian" is a light-sensitive eggplant cultivar, but its regulatory mechanism is unknown. Here, delphinidin-3-[4-(cis-p-coumaroyl)-rhamnosyl-glucopyranoside]-5-glucopyranoside and delphinidin-3-[4-(trans-p-coumaroyl)-rhamnosyl-glucopyranoside]-5-glucopyranoside were identified as the main anthocyanin components in Lanshan Hexian by ultra-performance liquid chromatography-tandem mass spectrometry. Three time points of anthocyanin accumulation, including the start point (0 day), fastest point (5 days), and highest point (12 day), were investigated by using ribonucleic acid sequencing and iTRAQ technology. The corresponding correlation coefficients of differentially expressed genes, and differentially expressed proteins were 0.6936, 0.2332, and 0.6672. Anthocyanin biosynthesis was a significantly enriched pathway, and CHI, F3H, 3GT, 5GT, and HY5 were regulated at both transcriptional and translational levels. Moreover, some transcription factors and photoreceptors may participate in light-induced anthocyanin biosynthesis like the known transcription factors MYB113 and TT8. The transient expression assay indicated that SmMYB35, SmMYB44, and a SmMYB86 isoform might involve in the light-induced anthocyanin biosynthesis pathway. Finally, a regulatory model for light-induced anthocyanin biosynthesis in eggplant was constructed. Our work provides a new direction for the study of the molecular mechanisms of light-induced anthocyanin biosynthesis in eggplant.

Cycloastragenol mediates activation and proliferation suppression in concanavalin A-induced mouse lymphocyte pan-activation model.

CONTEXT: Cycloastragenol (CAG) is a molecule isolated from various species in the genus Astragalus. Although the regulatory activity of Astragalus on immune system has been investigated, the effect of CAG on activated lymphocytes is poorly understood. OBJECTIVE: We aimed to biologically address the possible anti-inflammation potential of CAG on concanavalin A (Con A)-mediated mouse lymphocyte pan-activation model. MATERIALS AND METHODS: Mouse lymphocytes were obtained from spleens and subjected to Con A for 24 h. Herein, the cells were treated with different concentrations of CAG. Cell viability was assayed by MTT. Pretreated by CAG and stimulated by Con A, the expression of CD69 and CD25, Th1/Th2/Th17 cytokines, cell cycle, proliferation and intracellular Ca2+ concentration ([Ca2+]i) were analyzed by flow cytometry. RESULTS: The results declared that CAG significantly downregulated both CD69 and CD25 expressed on Con A activated CD3 + T cells' surface, as well as inhibiting proliferation of activated lymphocytes. In addition, CAG blocked the Con A-induced mitogenesis, exhibiting lymphocyte G0/G1-phase cell-cycle arrest with significant reduction of cells in S and G2/M phases. Meanwhile, pretreated by CAG, a significant decline in [Ca2+]i was observed. Furthermore, CAG significantly inhibited the production of Th1 cytokines IFN-γ, TNF, IL-2, Th2 cytokines IL-4, IL-6, IL-10 and Th17 cytokine IL-17 A on Con A-activated lymphocytes. CONCLUSION: Our results reinforce that CAG has important anti-inflammatory activity by inhibiting lymphocytes activation, proliferation and cytokines expression, and shows, that this effect may be related to reduction of overall intracellular Ca2+ overload.

Identification and characterization of CBL and CIPK gene families in eggplant (Solanum melongena L.).

Eggplant (Solanum melongena L.) is an edible vegetable cultivated and consumed worldwide. But the production of eggplant is significantly limited by the soil salinization in greenhouse cultivation system. The main ions are Na(+), Ca(2+), Mg(2+), K(+), Cl(-), and SO4 (2-) in the salty soils. Calcineurin B-like proteins (CBLs) are calcium sensors and control the affinities and activities of numerous ion transporters with CBL-interacting protein kinases (CIPKs). In this study, a total of 5 CBL and 15 CIPK genes from eggplant were identified first. The yeast two-hybrid (Y2H) assay and bimolecular fluorescence complementation (BiFC) assay demonstrated the interaction network between SmCBLs and SmCIPKs. Strikingly, some new CBL-CIPK complexes were found which have never been discovered in any other plant species. The expression level of each SmCBL or SmCIPK under 200 mM NaCl, low potassium (LK; 100 µM), high Mg with 20 mM MgCl2 and MgSO4 stresses were examined by quantitative real-time PCR (qRT-PCR) assay and these CBL and CIPK genes were found to respond to the four ion stresses differently. Interestingly, the differential expression level of SmCIPK3, -24 or -25 to Mg(2+) is higher than Na(+), and Cl(-) is higher than SO4 (2-). In addition, different magnesium salt can induce different response mechanisms in eggplant. In summary, this study provides insight into the characterization of CBLs and CIPKs in eggplant. It may be used in a novel biotechnological breeding program strategy to create new eggplant cultivars, leading to enhance different ion tolerance.

Novel insight into the mechanism underlying light-controlled anthocyanin accumulation in eggplant (Solanum melongena L.).

Eggplant is rich in anthocyanins, which are the major secondary metabolites and beneficial to human health. We discovered that the anthocyanin biosynthesis of eggplant cultivar 'Lanshan Hexian' was regulated by light. In this study, we isolated two blue light receptor genes, SmCRY1 and SmCRY2, and negative/positive anthocyanin regulatory factors SmCOP1 and SmHY5 from eggplant. In terms of transcript levels, SmCRY1, SmCRY2 and SmHY5 were up-regulated by light, while SmCOP1 was down-regulated. Subsequently, the four genes were functionally complemented in phenotype of corresponding mutants, indicating that they act as counterparts of Arabidopsis genes. Yeast two-hybrid and bimolecular fluorescence complementation assays showed that SmCRY1 and SmCRY2 interact with SmCOP1 in a blue-light-dependent manner. It also obtained the result that SmCOP1 interacts with SmHY5 and SmMYB1. Furthermore, using yeast one-hybrid assay, we found that SmHY5 and SmMYB1 both bind the promoters of anthocyanin biosynthesis structural genes (SmCHS and SmDFR). Taken together, blue-light-triggered CRY1/CRY2-COP1 interaction creates the condition that HY5 and MYB1 combine with the downstream anthocyanin synthesis genes (CHS and DFR) in eggplant. Our finding provides a new working model by which light controls anthocyanin accumulation in eggplant.

Review on emergency medical response against terrorist attack.

Terrorism is a global issue and a constant international threat. As a result, anti-terrorism and emergency response strategies are tasks of critical importance that have a direct impact on the national security of every country in the world. This paper reviews new characteristics of international anti-terrorism measures and offers an in-depth reflection on emergency medical response countermeasures; additionally, this paper presents the goals of related research, which include: 1) to present a model of a highly efficient medical response command; 2) to introduce the pre-planning phases of the emergency medical response; 3) to establish a response system capable of handling various types of terror attacks; 4) to promote anti-terrorism awareness to the general public and emphasize its prevention; and 5) to continue basic investigations into emergency medical responses for various types of terrorist attacks (for example, the classifications and characteristics of new injuries, pathophysiology, prevention and treatment of the resultant stress disorders, improved high-efficiency medical response measures and equipment, etc.).