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Objectives: Norovirus is associated with one-fifth of all gastroenteritis cases, but basic epidemiological data is lacking, especially in developing countries. As long-term surveillance on norovirus gastroenteritis is scarce in western China, this study aims to update the epidemiological knowledge of norovirus gastroenteritis and to characterize the genotypes of norovirus strains. Methods: Stool samples were collected from hospitalized children under 5 years old with gastroenteritis in Chengdu, China. All samples were tested for norovirus as well as rotavirus, sapovirus, enteric adenovirus, and astrovirus by real-time RT-PCR. RdRp and VP1 genes were sequenced in norovirus-positive samples to investigate viral phylogenies. Results: Of the 1181 samples collected from 2015 to 2019, 242 (20.5%) were positive for norovirus.Among norovirus-positive cases, 65 cases had co-infection with another virus; norovirus/enteric adenovirus was most frequently detected (50.8%, 33/65). The highest positive rate was observed in children aged 13–18 months (23.7%, 68/287). Norovirus infection peaked in autumn (36.6%, 91/249), followed by summer (20.3%, 70/345). Pearson correlation analysis showed significant correlation between the norovirus-positive rate and humidity (r = 0.773, P

Fibrosis is characterized by the excessive production of collagen and other extracellular matrix (ECM) components and represents a leading cause of morbidity and mortality worldwide. Previous studies of nonalcoholic steatohepatitis (NASH) with fibrosis were largely restricted to bulk transcriptome profiles. Thus, our understanding of this disease is limited by an incomplete characterization of liver cell types in general and hepatic stellate cells (HSCs) in particular, given that activated HSCs are the major hepatic fibrogenic cell population. To help fill this gap, we profiled 17,810 non-parenchymal cells derived from six healthy human livers. In conjunction with public single-cell data of fibrotic/cirrhotic human livers, these profiles enable the identification of potential intercellular signaling axes (e.g., ITGAV–LAMC1, TNFRSF11B–VWF and NOTCH2–DLL4) and master regulators (e.g., RUNX1 and CREB3L1) responsible for the activation of HSCs during fibrogenesis. Bulk RNA-seq data of NASH patient livers and rodent models for liver fibrosis of diverse etiologies allowed us to evaluate the translatability of candidate therapeutic targets for NASH-related fibrosis. We identified 61 liver fibrosis-associated genes (e.g., AEBP1, PRRX1 and LARP6) that may serve as a repertoire of translatable drug target candidates. Consistent with the above regulon results, gene regulatory network analysis allowed the identification of CREB3L1 as a master regulator of many of the 61 genes. Together, this study highlights potential cell–cell interactions and master regulators that underlie HSC activation and reveals genes that may represent prospective hallmark signatures for liver fibrosis.

Glioblastoma (GBM) is a prevalent and highly lethal form of glioma, with rapid tumor progression and frequent recurrence. Excessive outgrowth of pericytes in GBM governs the ecology of the perivascular niche, but their function in mediating chemoresistance has not been fully explored. Herein, we uncovered that pericytes potentiate DNA damage repair (DDR) in GBM cells residing in the perivascular niche, which induces temozolomide (TMZ) chemoresistance. We found that increased pericyte proportion correlates with accelerated tumor recurrence and worse prognosis. Genetic depletion of pericytes in GBM xenografts enhances TMZ-induced cytotoxicity and prolongs survival of tumor-bearing mice. Mechanistically, C-C motif chemokine ligand 5 (CCL5) secreted by pericytes activates C-C motif chemokine receptor 5 (CCR5) on GBM cells to enable DNA-dependent protein kinase catalytic subunit (DNA-PKcs)-mediated DDR upon TMZ treatment. Disrupting CCL5-CCR5 paracrine signaling through the brain-penetrable CCR5 antagonist maraviroc (MVC) potently inhibits pericyte-promoted DDR and effectively improves the chemotherapeutic efficacy of TMZ. GBM patient-derived xenografts with high CCL5 expression benefit from combined treatment with TMZ and MVC. Our study reveals the role of pericytes as an extrinsic stimulator potentiating DDR signaling in GBM cells and suggests that targeting CCL5-CCR5 signaling could be an effective therapeutic strategy to improve chemotherapeutic efficacy against GBM.

The Cd accumulating difference in edible parts of leafy vegetables were involvement with the cultivar-dependent oxidation resistance of plants. According to the transcriptomic results, the total expression of the 4 major antioxidase genes in Cd pollution-safe cultivars (Cd-PSCs) under control condition were higher than in high-Cd-accumulating cultivars (HCCs) of the 4 tested leafy vegetables, indicating the cultivar-dependent high oxidation resistance of Cd-PSCs. Simultaneously, expression of superoxide dismutase (SOD) and peroxidase (POD) genes in Cd-PSCs were all higher than in HCCs under control condition. The real detection of SOD activities in Cd-PSCs under control condition were higher than in HCCs, which verified the transcriptomic results. What’s more, there were no uniform response mechanisms of antioxidase genes to cope with Cd stress among the 4 tested leafy vegetables, even 3 of them were native from the same genus Brassica. Meanwhile, expression of the 4 major antioxidase genes were difference from genus, such as the polyphenol oxidase (PPO) genes were hardly expression in genus Brassica. These results would contribute to the understanding the inherent difference between Cd-PSCs and HCCs, and the cultivar-dependent Cd oxidation resistance related to Cd-PSCs might a easy signal for in vitro catching and screening.

Aberrant activation of Wnt/β-catenin signaling by dysregulated post-translational protein modifications, especially ubiquitination is causally linked to cancer development and progression. Although Lys48-linked ubiquitination is known to regulate Wnt/β-catenin signaling, it remains largely obscure how other types of ubiquitination, such as linear ubiquitination governs its signaling activity. The expression and regulatory mechanism of linear ubiquitin chain assembly complex (LUBAC) on Wnt/β-catenin signaling was examined by immunoprecipitation, western blot and immunohistochemical staining. The ubiquitination status of β-catenin was detected by ubiquitination assay. The impacts of SHARPIN, a core component of LUBAC on malignant behaviors of gastric cancer cells were determined by various functional assays in vitro and in vivo. Unlike a canonical role in promoting linear ubiquitination, SHARPIN specifically interacts with β-catenin to maintain its protein stability. Mechanistically, SHARPIN competes with the E3 ubiquitin ligase β-Trcp1 for β-catenin binding, thereby decreasing β-catenin ubiquitination levels to abolish its proteasomal degradation. Importantly, SHARPIN is required for invasiveness and malignant growth of gastric cancer cells in vitro and in vivo, a function that is largely dependent on its binding partner β-catenin. In line with these findings, elevated expression of SHARPIN in gastric cancer tissues is associated with disease malignancy and correlates with β-catenin expression levels. Our findings reveal a novel molecular link connecting linear ubiquitination machinery and Wnt/β-catenin signaling via SHARPIN-mediated stabilization of β-catenin. Targeting the linear ubiquitination-independent function of SHARPIN could be exploited to inhibit the hyperactive β-catenin signaling in a subset of human gastric cancers.

The heavy metal poisoning in humans and fish represents a significant global problem. Copper (Cu), as an essential micronutrient in human and animal metabolism, often accumulates excessively in aquatic environment. The microbial floc is rich in a variety of probiotics and bioactive compounds, which has been documented to have the functions of antioxidant and immunoenhancement. A 64-day experiment was conducted to investigate the protective effects and potential mechanisms of dietary supplementation of microbial floc and Cu exposure on inflammatory response, oxidative stress, intestinal apoptosis and barrier dysfunction in Rhynchocypris lagowski Dybowski. A total of four hundred fifty R. lagowski were fed five experimental diets containing graded levels of microbial floc from 0% to 16% (referred to as B0, B4, B8, B12 and B16, respectively) in the first 60 days, and 96 h of acute copper exposure test was carried out in the last four days. The results showed that microbial floc exerted significant alleviative effects by preventing alterations in the levels of bioaccumulation, caspase3, caspase8, caspase9, malondialdehyde and interleukin-6, improving the activities of lysozyme, complement C3, complement C4, immunoglobulin M, alkaline phosphatase, heat shock protein 70, heat shock protein 90 and glutathione peroxidase, catalase, superoxide dismutase, total antioxidant capacity. In addition, microbial floc assisted in regulating the expression of NF-κB/Nrf2 signaling molecule genes, including NF-κB, TNF-α, IL-1β, IL-8, IL-10, TGF-β, Keap1, Nrf2, Maf, HO-1, CAT, CuZn-SOD, GCLC and GPX. Overall, our results suggest that dietary supplementation with of microbial floc can alleviate copper-induced inflammation, oxidative stress, intestinal apoptosis and barrier dysfunction in R. lagowski. A suitable supplementation level of approximately 12% microbial floc is recommended in the present study.

Purpose Clear cell renal cell carcinoma(ccRCC) is the most common type of renal cell carcinoma. While it is curable when detected at an early stage, some patients presented with advanced disease have poor prognosis. We aimed to identify key genes and miRNAs associated with clinical prognosis in ccRCC. Methods The microarray datasets were obtained from the Gene Expression Omnibus database. Differentially expressed genes (DEGs) and differentially expressed miRNAs (DEMs) were analyzed by using GEO2R. Then, Functional enrichment analysis was performed using the DAVID. A retrospective series of 254 ccRCC patients with complete clinical information was included in this study. Kaplan-Meier analysis and multivariate cox regression analysis were used for prognostic analysis. Wound healing assay and transwell assay were designed to evaluate the migration and invasion ability of ccRCC cell lines. Results miRNA-18a was identified to be related with prognosis of ccRCC by using Kaplan-Meier analysis and multivariate cox regression analysis demonstrated that the prognostic value of miRNA-18a was independent of clinical features. Further studies showed that up-regulation of miRNA-18a had a positive effect on migration and invasion of ccRCC cells. The target gene (HIF1A) of the miRNA-18a was predicted by using the miRPathDB database. The transcription factors of DEGs were identified by using the i-cisTarget. Luckily, HIF1A was found to be one of the transcription factors of DEGs. Among these DEGs, PVT1 may be regulated by HIF1A and be related with prognosis of ccRCC. Finally, validation of miRNA18a/HIF1A/PVT1 pathway was checked via reverse transcription-polymerase chain reaction (RT-PCR) assay in both cell lines and clinical tumor samples. Conclusion Our research revealed that miRNA18a/HIF1A/PVT1 pathway might play a crucial role in ccRCC progression, providing novel insights into understanding of ccRCC molecular mechanisms. Importantly, miRNA-18a could serve as a potential diagnostic biomarker and therapeutic targets for ccRCC patients.

A highly efficient di-C-glycosyltransferase GgCGT was discovered from the medicinal plant Glycyrrhiza glabra. GgCGT catalyzes a two-step di-C-glycosylation of flopropione-containing substrates with conversion rates of >98%. To elucidate the catalytic mechanisms of GgCGT, we solved its crystal structures in complex with UDP-Glc, UDP-Gal, UDP/phloretin, and UDP/nothofagin, respectively. Structural analysis revealed that the sugar donor selectivity was controlled by the hydrogen-bond interactions of sugar hydroxyl groups with D390 and other key residues. The di-C-glycosylation capability of GgCGT was attributed to a spacious substrate-binding tunnel, and the G389K mutation could switch di- to mono-C-glycosylation. GgCGT is the first di-C-glycosyltransferase with a crystal structure, and the first C-glycosyltransferase with a complex structure containing a sugar acceptor. This work could benefit the development of efficient biocatalysts to synthesize C-glycosides with medicinal potential.

Lamiophlomis rotata (Benth.) Kudo is a herbaceous plant of the family Lamiaceae, subfamily Lamioideae. Approximately, 127 chemical constituents have been isolated and identified from L. rotata, including iridoids, flavonoids, phenylethanoid glycosides, polysaccharides, and organic acids. These chemical constituents have extensive pharmacological properties, which include anti-nociceptive, haemostatic, anti-inflammatory, anti-tumour, immunomodulatory, antioxidant, and cardio-protective activities. Documentation of its historical use in traditional medicine and contemporary phytochemical and pharmacological research indicate that L. rotata has significant potential in therapeutic and health care applications. Both whole extracts and individual chemical components isolated from this plant exhibit a wide range of biological activities that warrant further investigation. These investigations can be assisted by careful review of existing traditional knowledge from diverse cultural backgrounds. A new search for chemical and biological markers and reinforced protection of the germplasm resources of L. rotata are also important to ensure targeted and sustainable use of this medicinal resource. The aim of this review was to provide comprehensive information on the botanical characteristics, traditional uses, ethnopharmacology, chemical and pharmacological properties, toxicity profile, and conservation status of L. rotata, to improve understanding of its mechanisms of action so that novel therapeutic agents may be developed from this plant.

Microsatellite instability (MSI) has been approved as a pan-cancer biomarker for immune checkpoint blockade (ICB) therapy. However, current MSI identification methods are not available for all patients. We proposed an ensemble multiple instance deep learning model to predict microsatellite status based on histopathology images, and interpreted the pathomics-based model with multi-omics correlation. Methods: Two cohorts of patients were collected, including 429 from The Cancer Genome Atlas (TCGA-COAD) and 785 from an Asian colorectal cancer (CRC) cohort (Asian-CRC). We established the pathomics model, named Ensembled Patch Likelihood Aggregation (EPLA), based on two consecutive stages: patch-level prediction and WSI-level prediction. The initial model was developed and validated in TCGA-COAD, and then generalized in Asian-CRC through transfer learning. The pathological signatures extracted from the model were analyzed with genomic and transcriptomic profiles for model interpretation. Results: The EPLA model achieved an area-under-the-curve (AUC) of 0.8848 (95% CI: 0.8185-0.9512) in the TCGA-COAD test set and an AUC of 0.8504 (95% CI: 0.7591-0.9323) in the external validation set Asian-CRC after transfer learning. Notably, EPLA captured the relationship between pathological phenotype of poor differentiation and MSI (P < 0.001). Furthermore, the five pathological imaging signatures identified from the EPLA model were associated with mutation burden and DNA damage repair related genotype in the genomic profiles, and antitumor immunity activated pathway in the transcriptomic profiles. Conclusions: Our pathomics-based deep learning model can effectively predict MSI from histopathology images and is transferable to a new patient cohort. The interpretability of our model by association with pathological, genomic and transcriptomic phenotypes lays the foundation for prospective clinical trials of the application of this artificial intelligence (AI) platform in ICB therapy.

Distribution of chemical forms of manganese(II) (Mn(II)) on plant roots may affect Mn(II) absorption by plants and toxicity of Mn(II) to plants at its high level. The chemical forms of Mn(II) on soybean roots were investigated to determine the main factors that affect their distribution and relationship with Mn(II) plant toxicity. Fresh soybean roots were reacted with Mn(II) in solutions, and Mn(II) adsorbed on the roots was differentiated into exchangeable, complexed, and precipitated forms through sequential extraction with KNO3, EDTA, and HCl. The exchangeable Mn(II) content on the roots was the highest, followed by the complexed and precipitated Mn(II) contents. Mn(II) toxicity to the roots was greater at pH 5.5 than at pH 4.2 due to the larger amount of exchangeable Mn(II) at higher pH. The cations Al3+, La3+, Ca2+, Mg2+, and NH4+ competed with Mn(II) for cation exchange sites on the root surfaces and thus reduced exchangeable Mn(II) on the roots, in the order Al3+, La3+ > Ca2+, Mg2+ > NH4+. Al3+ and La3+ at 100 μmol L−1 decreased exchangeable Mn(II) by 80% and 79%, respectively, and Ca2+ and Mg2+ at 1 mmol L−1 decreased exchangeable Mn(II) by 51% and 73%, respectively. Organic anions oxalate, citrate, and malate reduced free Mn(II) concentration in solution through formation of complexes with Mn(II), efficiently decreasing exchangeable Mn(II) on the roots; the decreases in exchangeable Mn(II) on the roots were 30.9%, 19.7%, and 10.9%, respectively, which was consistent with the complexing ability of these organic anions with Mn(II). Thus, exchangeable Mn(II) was the dominant form of Mn(II) on the roots and responsible for Mn(II) toxicity to plants. The coexisting cations and organic anions reduced the exchangeable Mn(II) content, and thus they could alleviate Mn(II) toxicity to plants on acid soils.

Cancer metastasis, a leading cause of death in patients, is associated with aberrant expression of epigenetic modifiers, yet it remains poorly defined how epigenetic readers drive metastatic growth and whether epigenetic readers are targetable to control metastasis. Here, we report that bromodomain-containing protein 4 (BRD4), a histone acetylation reader and emerging anticancer therapeutic target, promotes progression and metastasis of gastric cancer. The abundance of BRD4 in human gastric cancer tissues correlated with shortened metastasis-free gastric cancer patient survival. Consistently, BRD4 maintained invasiveness of cancer cells in vitro and their dissemination at distal organs in vivo. Surprisingly, BRD4 function in this context was independent of its putative transcriptional targets such as MYC or BCL2, but rather through stabilization of Snail at posttranslational levels. In an acetylation-dependent manner, BRD4 recognized acetylated lysine 146 (K146) and K187 on Snail to prevent Snail recognition by its E3 ubiquitin ligases FBXL14 and β-Trcp1, thereby inhibiting Snail polyubiquitination and proteasomal degradation. Accordingly, genome-wide transcriptome analyses identified that BRD4 and Snail regulate a partially shared metastatic gene signature in gastric cancer cells. These findings reveal a noncanonical posttranscriptional regulatory function of BRD4 in maintaining cancer growth and dissemination, with immediate translational implications for treating gastric metastatic malignancies with clinically available bromodomain inhibitors. Significance: These findings reveal a novel posttranscriptional regulatory function of the epigenetic reader BRD4 in cancer metastasis via stabilizing Snail, with immediate translational implication for treating metastatic malignancies with clinically available bromodomain inhibitors.

Flower color variegation has been observed in many plant species. However, pink flowers on the white-blooming hawthorn trees found by our group earlier have never been reported. To better understand the differentially expressed genes (DEGs) in variegated hawthorn flowers, white and pink flowers at different developmental stages (S1 and S2) underwent transcriptome sequencing separately. Approximately 34.28 Gb of high-quality data were obtained and assembled into 100,013 unigenes with an average length of 706.93 bp. These unigenes were further subjected to functional annotation and biochemical pathway analysis, and DEGs of two types of hawthorn flowers at different developmental stages were studied. Based on the enrichment analysis of DEGs, eight anthocyanin-modified enzyme genes or other enzyme genes that indirectly affect anthocyanin synthesis (5AT, 3GGT, and AI, β-Glu, two Aux/IAAs, two PODs), eight structural genes (UFGT, DFR, CHI, two F3Hs, and three PALs), and three transcription factors (one MYB and two bHLHs) were also identified. We randomly selected 15 genes, and the trends in the expression levels of these genes in the organs of white and pink flowers at different developmental stages were verified by quantitative real-time PCR. Mass sequence data obtained by RNA-seq of variegated hawthorn flowers provided basic sequence information and a unique opportunity to uncover the genetic mechanisms underlying flower color variegation.

Unlike animals, few vascular plants are known to spontaneously generate rhythmic electrical signals. In plants electrical signals triggered in response to external environmental changes are of physiological significance. However, plant electrical signals require experimental skill to measure, are stimulus-pattern dependent, and show poor quantitative reproducibility, thus their existence has been largely ignored by researchers. We developed a non-invasive and high-throughput light-induced in situ method to mine the information conveyed by reproducible bioelectrical activity in plants. Stable periodic electrical potential changes in wheat leaves induced by illumination–darkness cycles enabled selection of wave features and specific classification algorithms were used to enable rapid distinction of salt-tolerant and salt-sensitive cultivars. Chlorophyll fluorescence data also strongly supported the association of rhythmic electrical potential with salt sensitivity in wheat leaves. The results show that the electrophysiological phenotype represented by plant rhythmic electrical potential is a novel candidate for phenotypic analyses. The method will also enable rapid assessment of the response and resistance to abiotic stresses of crop seedlings.

Background: Our previous study confirmed the beneficial effects of chestnut polysaccharides on the spermatogenesis process, but the exact mechanism is not clear. Several studies have demonstrated the importance of a balanced gut microbiota in maintaining normal reproductive function. In this study, we investigated the biological functions of chestnut polysaccharides from the perspective of gut microbiota function, expecting to elaborate the specific mechanism of chestnut polysaccharides beneficial to spermatogenesis.Results: Compared with the Vehicle group, germ cell quantity, intestinal structure and tight junction of intestinal tissue were altered in the Busulfan-treated mice, and the intestinal microbiota was also disturbed at several levels, including phylum and genus. The Firmicutes was predominant bacteria in all group followed by Proteobacteria, Bacteroidetes, Actinobacteria, Tenericutes, Cyanobacteria and unidentified-Bacteria, however, the composition of those gut microbiota changes caused by different treatment. Busulfan disturbed the homeostasis of the intestinal microenvironment. Chestnut polysaccharides could rescue this aberrance and improve spermatogenesis via steroid hormone synthesis process. Conclusions: Chestnut polysaccharides can change gut structure and tight junctions to rescue Busulfan-impaired spermatogenesis by altering the composition of different groups of gut microbes, a rescue process that is most likely related to hormone synthesis processes. It will provide a new insight for treating the male-related infertility.

Colorectal cancer (CRC) is a common malignant tumor of the digestive tract and one of the leading causes of cancer-associated mortality. Secreted phosphoprotein-1 (SPP-1) is overexpressed in CRC and promotes cancer progression, but the underlying mechanisms underlying SPP-1 function remain unclear. The present study aimed to explore the effects of Wnt/β-catenin signaling in SPP-1-induced CRC progression. The expression patterns of SPP-1 in CRC tissues were examined using reverse transcription-quantitative (RT-q)PCR, western blotting and immunohistochemistry. SPP-1 expression in cells was assessed using RT-qPCR and western blotting. Cell-Counting Kit-8, flow cytometry and tumor-burdened mice experiments were used to determine cell proliferation, apoptosis and in vivo tumor formation abilities. The results showed that SPP-1 expression was markedly elevated in CRC tissues and cells compared with that in normal colorectal tissues and cells. High expression of SPP-1 was associated with advanced clinical process and low overall survival rate in patients with CRC. Besides, SPP-1 could interact with β-catenin and positively regulated β-catenin protein expression, and enhanced its nuclear accumulation. Moreover, SPP-1-upregulation significantly enhanced cell proliferation and in vivo tumor formation ability, and reduced apoptosis, whereas these effects were all abolished when β-catenin was silenced. Overall, the present study revealed that SPP-1 promoted the progression of CRC in a β-catenin-dependent manner.

Background Our previous studies demonstrated that the administration of crude Polysaccharide from Panax notoginseng (CPPN) can effectively prolong the lifespan of tumor-bearing mice via boosting the host immune system as well as weak cytotoxicity against hepatocellular carcinoma (HCC). In the present study, Neutral Polysaccharide (NPPN) were further purified from crude polysaccharide isolated from panax notoginseng. The effects of NPPN on the immune function and hematopoietic function of mice with low immunity and myelosuppression induced by cyclophosphamide (CTX) were investigated. The effect of NPPN combined with CTX on the tumor inhibition rate of the H22 tumor-bearing mice and the impact of NPPN on the proliferation of H22 liver cancer cells in vitro were investigated. Methods CPPN was obtained by water extraction and alcohol precipitation method, and further purified by DEAE Sepharose Fast Flow ion exchange resin column. NPPN was added to the immunosuppressed with myelosuppression mice induced by CTX. Thymus index, spleen index, lymphocyte proliferation stimulation index by adding of concanavalin A, determination of serum hemolysin, NK cell activity assay, mice carbon clearance experiment, blood count tests were detected. The tumor inhibition rate of the H22 tumor-bearing mice treated with NPPN combined with CTX was recorded. Results NPPN and 4 kinds of acid polysaccharide from Panax notoginseng (APPN) were successfully isolated from the CPPN by DEAE Sepharose Fast Flow ion exchange resin column. NPPN inhibited the growth of H22 cells and significantly increase the tumor inhibition rate of the H22 tumor-bearing mice combined with CTX. The elevation of the cellular and humoral immunity levels as well as a variety of blood count tests indicators of immunosuppressive with myelosuppression mice may contribute to the antitumor activity of NPPN. Conclusion NPPN has a potential antitumor activity for the treatment of liver cancer combined with cyclophosphamide.

Gene-expression programs define shared and species-specific phenotypes, but their evolution remains largely uncharacterized beyond the transcriptome layer 1 . Here we report an analysis of the co-evolution of translatomes and transcriptomes using ribosome-profiling and matched RNA-sequencing data for three organs (brain, liver and testis) in five mammals (human, macaque, mouse, opossum and platypus) and a bird (chicken). Our within-species analyses reveal that translational regulation is widespread in the different organs, in particular across the spermatogenic cell types of the testis. The between-species divergence in gene expression is around 20% lower at the translatome layer than at the transcriptome layer owing to extensive buffering between the expression layers, which especially preserved old, essential and housekeeping genes. Translational upregulation specifically counterbalanced global dosage reductions during the evolution of sex chromosomes and the effects of meiotic sex-chromosome inactivation during spermatogenesis. Despite the overall prevalence of buffering, some genes evolved faster at the translatome layer-potentially indicating adaptive changes in expression; testis tissue shows the highest fraction of such genes. Further analyses incorporating mass spectrometry proteomics data establish that the co-evolution of transcriptomes and translatomes is reflected at the proteome layer. Together, our work uncovers co-evolutionary patterns and associated selective forces across the expression layers, and provides a resource for understanding their interplay in mammalian organs.

In the present study, antagonistic activity of bacterial strain BS-Z15, was evaluated against Verticillium dahlia. The fermented broth of BS-Z15 inhibited the growth of Verticillium dahliae. The genome of strain BS-Z15 had a total size of 4,068,702 base pairs and contained 4318 genes, of which 4196 are coding sequences and 122 are non-coding RNA. Among these genes, nine genomic islands, 86 tRNAs, 13 sRNAs, and one prophage was determined. With the help of annotation databases, most unigene functions were identified. At the same time, genomic comparison between BS-Z15 and 12 Bacillus members showed that the genes of BS-Z15 were closely related to the Bacillus group, and were conserved between the two groups, including most of the genes associated with fungal antagonism. BS-Z15 contains genes involved in a variety of antagonistic mechanisms, including genes encoding or synthesizing mycosubtilin, chitinases (but not CHIA and CHIB), glycoside hydrolases, iron nutrients, and antibiosis. However, it only contained the complete mycosubtilin- and bacilibactin-related operators in the reported main antifungal gene cluster of B. subtilis. Mycosubtilin and bacilibactin may be the main active antifungal substance. Besides, some genes could encode products related to biofilm production, which may be related to the colonization ability of the strain in plant rhizospheres. The complete genome of B. subtilis BS-Z15 provided new insights into the potential metabolites it produces related to its biocontrol activity.

Systemic lupus erythematosus (SLE) is a genetically complex autoimmune disease. Despite the significant progress made in identifying susceptibility genes for SLE, the genetic architecture of the disease is far from being understood. In this study, we set to replicate a number of suggestive association signals found in genome-wide association studies (GWASs) in additional independent cohorts. Replication studies were performed on Han Chinese cohorts from Hong Kong and Anhui, involving a total of 2,269 cases and 5,073 controls. We identified a missense variant in IRF3 (rs7251) reaching genome-wide significance through a joint analysis of GWAS and replication data (OR = 0.876, P = 4.40E-08). A significant correlation was observed between rs7251 and lupus nephritis (LN) by subphenotype stratification (OR = 0.785, P = 0.0128). IRF3 is a key molecule in type I interferon production upon nucleic acid antigen stimulations and may inhibit regulatory T cell differentiation. Further elucidation of the mechanism of this association could help us better understand the pathogenesis of SLE.

As a further exploration of the asymmetrically substituted diiron models for the active site of [FeFe]-hydrogenases, two new types of small bite-angle aminodiphosphine [(Ph2P)2NR; denoted as PNP in this study]-chelated diiron N-phenyl-aza- and ethanedithioate complexes Fe2(μ-xdt)(CO)4{κ2-(Ph2P)2NR} (1a–1e) and (2a–2e), respectively, were successfully synthesized by the carbonyl substitution reactions of all-carbonyl diiron complexes Fe2(μ-xdt)(CO)6 (xdt = SCH2N(Ph)CH2S (adtNPh) and SCH2CH2S (edt)) with PNP (PNP = (Ph2P)2NR, R = CMe3, CH2CHMe2, (CH2)3Me, (CH2)3Si(OEt)3, and (CH2)3NMe2) in the presence of Me3NO·2H2O or UV irradiation. All the new complexes obtained above have been well characterized by elemental analysis, FT-IR, NMR spectroscopy, and particularly for 1a, 1b, 2a, and 2d by single-crystal X-ray diffraction analysis. By comparison, 31P{1H} NMR and X-ray crystallographic studies have clearly revealed that the change of the dithiolate bridge from adtNPh to edt has a significant influence on the ...

To explore the possible roles of metallothioneins (MTs) played in cadmium (Cd) accumulation of water spinach, three IaMT genes, IaMT1, IaMT2 and IaMT3 in a high-shoot-Cd (T308) and a low-shoot-Cd accumulation cultivar (QLQ) were cloned, characterized, and quantitated. Gene expression analysis suggested that the expression of the IaMTs was differentially regulated by Cd stress in different cultivars, and T308 showed higher MTs expression overall. Furthermore, only shoot IaMT3 expression was cultivar dependent among the three IaMTs. Antioxidant analysis showed that the high production of IaMTs in T308 should be associated with its high oxidation resistance. The role of IaMTs in protecting against Cd toxicity was demonstrated in vitro via recombinant E. coli strains. The results showed that IaMT1 correlated with neither Cd tolerance nor Cd accumulation of E. coli, while IaMT2 conferred Cd tolerance in E. coli, IaMT2 and IaMT3 increased Cd accumulation in E. coli. These findings help to clarify the roles of IaMTs in Cd accumulation, and increase our understanding of the cultivar-dependent Cd accumulation in water spinach.

This study evaluated whether tumor expression of programmed death ligand 1 (PD-L1) could predict the response of EGFR-mutated NSCLC to EGFR tyrosine kinase inhibitor (TKI) therapy.We retrospectively evaluated patients who received EGFR-TKIs for advanced NSCLC at the Guangdong Lung Cancer Institute between April 2016 and September 2017 and were not enrolled in clinical studies. The patients' EGFR and PD-L1 statuses were simultaneously evaluated.Among the 101 eligible patients, strong PD-L1 expression significantly decreased objective response rate, compared with weak or negative PD-L1 expression (35.7% versus 63.2% versus 67.3%, p = 0.002), and shortened progression-free survival (3.8 versus 6.0 versus 9.5 months, p0.001), regardless of EGFR mutation type (19del or L858R). Furthermore, positive PD-L1 expression was predominantly observed among patients with de novo resistance rather than acquired resistance to EGFR-TKIs (66.7% versus 30.2%, p = 0.009). Notably, we found a high proportion of PD-L1 and cluster of differentiation 8 (CD8) dual-positive cases among patients with de novo resistance (46.7%, 7 of 15). Finally, one patient with de novo resistance to EGFR-TKIs and PD-L1 and CD8 dual positivity experienced a favorable response to anti-programmed death 1 therapy.This study revealed the adverse effects of PD-L1 expression on EGFR-TKI efficacy, especially in NSCLC patients with de novo resistance. The findings indicate the reshaping of an inflamed immune phenotype characterized by PD-L1 and CD8 dual positivity and suggest potential therapeutic sensitivity to programmed death 1 blockade.

Taxus yunnanensis (Yew) is known for natural anticancer metabolite paclitaxel (Taxol) and its biosynthesis pathway in yew species still needs to be completely elucidated. In the current study, productions of paclitaxel and 10-DAB III from three different tissues (needle, branch, and root) of T. yunnanensis wild type (WT) and two new cultivars Zhongda-1 (Zd1) and Zhongda-2 (Zd2) were determined, and significant tissue differences in contents of the taxanes were observed among the three experimental lines. The much higher 10-DAB III and lower paclitaxel contents in needle of Zd2 when compared with that of Zd1 indicated the low conversion from 10-DAB III to paclitaxel in the needle of Zd2. In order to uncover the mechanisms of the tissue-specific biosynthesis of the taxanes, transcriptome analysis of cultivar Zd2 was conducted, and the previously reported transcriptome data of Zd1 and WT were used to perform a comparison. The enhancement of TDAT and T10βH side biosynthetic pathway in roots of Zd2 in early taxane synthesis might lead to the biosynthesis of other toxoids, while the preference of T13αH route in the needle and branch of Zd2 was mainly responsible for the tissue-specific reinforced biosynthesis of 10-DAB III and paclitaxel in Zd2. Different from Zd1, the tissue-specific pattern of paclitaxel biosynthesis genes in Zd2 was similar to WT. However, the lower transcript abundance of final steps genes (TBT, DBAT, BAPT, and DBTNBT) of the paclitaxel biosynthesis pathway in Zd2 than in Zd1 might further promote 10-DAB III accumulation in Zd2.

Tumor growth is fueled by subset of cells with stem cell properties (Cancer stem cells, CSCs). While persistent activation of Wnt/β-catenin signaling confers CSC properties, it remains unclear how epigenetic modifications regulate Wnt target genes to dictate their self-renewal. Here, we report a novel Wnt-responsive epigenetic switch for CSC maintenance through activating the stem cell transcription factor ASCL2 in gastric carcinoma (GC). We characterize ASCL2-expressing (ASCL2+) GC cells as a subset of Wnt-responsive CSCs that depend on ASCL2 for self-renewal. High-throughput RNAi screening uncovers that the histone methyltransferase SMYD3 determines H3K4me3 status at the ASCL2 locus to promote ASCL2 expression. Moreover, SMYD3 may be transcriptionally activated by the β-catenin/TCF4 complex, indicating that the SMYD3-ASCL2 axis may be an integral component of Wnt signaling. Consistently, SMYD3 maintains self-renewal and tumorigenicity of ASCL2+ CSCs largely through inducing ASCL2. Clinically, overexpression of SMYD3 and ASCL2 are associated with malignant progression and poor patient outcomes in GC. Together, these findings define a Wnt-responsive CSC pathway that could be exploited to identify essential regulators of the signaling output, and reveal SMYD3 as an epigenetic target for eliminating CSCs in human cancers.

Introduction Inhibition of programmed cell death-1 (PD-1) and its ligand programmed death ligand 1 (PD-L1) by using an immune checkpoint inhibitor has emerged as a promising immunotherapy for NSCLC. The correlation of PD-L1 expression in tumor cells with treatment outcomes has been reported in many pivotal trials; however, the relationship remains unclear. Here, we demonstrate that those patients with both high density of PD-1–positive CD8 and PD-L1–positive CD4-positive CD25-positive (PD-1 hi PD-L1 hi ) regulatory T cells (Tregs) have a better response to PD1/PD-L1 blockade. Methods In our study between April 1, 2014, and May 30, 2017, a total of 73 NSCLC peripheral blood samples and fresh tumor specimens were collected for study. Of these, 42 large (10-mm 3 ) fresh tumor specimens were obtained from surgical procedures and checked for expression of immunology biomarkers, including PD-L1, PD-1, CD8, CD4, and CD25, in tumor cells and tumor-infiltrating lymphocytes (TILs) by flow cytometry, immunohistochemistry, and immunofluorescence (IF). Moreover, 31 small biopsy specimens from patients who received immunotherapy (pembrolizumab or nivolumab) were analyzed by immunohistochemistry and IF. The correlation between flow cytometry and IF detected for TILs' density was evaluated by Spearman's rank correlation test; the primary end point was progression-free survival. For the PD-1/PD-L1 blockade assay, the TILs and peripheral blood mononuclear CD8 T cells were cultured (1×10 5 per well) with anti–PD-1 (clone MIH4), anti–PD-L1 (clone MIH1). The cytotoxic activity of TILs in killing NSCLC cells after stimulation by anti–PD-1 and anti–PD-L1 was measured by a conventional 51 Cr release assay. Results We first identified a population of high–PD-L1–expressing CD25-positive CD4-positive T cells (PD-L1 hi Tregs) in the tumor microenvironment. The frequency of PD-L1 hi Tregs was higher in tumor tissue (mean 48.6 ± 14.3% in CD25-positive CD3-positive CD4-positive T cells) than in blood (mean 35.4 ± 10.2% in CD25-positive CD3-positive CD4-positive T cells) and normal tissue (mean 38.6 ± 9.7% in CD25-positive CD3-positive CD4-positive T cells) ( p hi Tregs was positively correlated with PD-1–positive CD8 in Tregs. In addition, the TILs from these patients (PD-1 hi PD-L1 hi ) showed PD-1/PD-L1 pathway dependence and could induce a greater killing effect of TILs by PD-1/PD-L1 blockade treatment. The patients with PD-L1–positive NSCLC with PD-1 hi PD-L1 hi TILs showed a better clinical outcome than those with a low frequency of PD-1 hi CD8 or PD-L1 hi Tregs (median progression-free survival not reached versus 2 months). Conclusions Our findings suggested that the density of PD-L1–positive CD4-positive CD25-positive Tregs in the tumor microenvironment can serve as a diagnostic factor to supplement PD-L1 expression in tumor cells and predict the response to PD-1/PD-L1 blockade immunotherapy in NSCLC.

Pure Cu nanowires as catalyst were prepared by electrochemical deposition and were used in CO2 hydrogenation to methanol. The active sites of the Cu based catalyst were discussed. The performance and structural development of the catalyst were observed during CO2 hydrogenation. A mechanism for the deactivation of the catalyst was discussed. The key factors that affect the deactivation of the catalyst were found. Cu nanowire sample was characterized by SEM, EDS, XRD, and BET. The results show that Cu nanowires have very high sintering resistance and catalytic stability. This helps to develop high performance catalysts. The changes in the grain size, SEM morphology and catalytic properties of the sample during CO2 hydrogenation show that the migration of the Cu atoms on the surface of the Cu nanowires can occur. Continuous migration of Cu atoms and sintering of Cu grains can lead to flow blockage in gas channels. The gas channel flow blockage or the sintering of Cu grains can lead to deactivation of the catalyst. However, the shape of catalytic performance curve indicates that the main reason for the deactivation of the catalyst is the gas channel flow blockage.

Microalgae have important applications in the aquaculture industry due to their remarkable nutritional value, especially the high contents of carotenoids. In this study, five microalgae, namely Spirulina sp., Dunaliella salina, Haematococcus pluvialis, Isochrysis galbana and Myrmecia incisa were investigated for their beneficial effects on Artemia nauplii, their predator in the food chain. For H. pluvialis (rich in astaxanthin), I. galbana (rich in fucoxanthin) and M. incisa (limited carotenoids but rich in arachidonic acid), they were cultured under either normal or stressful condition to produce the algal samples containing different amounts of carotenoids or fatty acids. Results showed that through feeding, the carotenoids in microalgae can be efficiently transferred to Artemia nauplii. In comparison with fatty acids, carotenoids made more contribution to the improved growth and health conditions of Artemia, leading to better performances in survival rate, average body length, total effective body length and total antioxidant capacity. Besides, the immunity of Artemia nauplii was also enhanced, as evidenced by the strong resistance against Vibrio parahaemolyticus-induced infection. Findings of the present study clearly supported the benefits of carotenoid-rich microalgae for Artemia nauplii, providing a feasible microalgae-based strategy to produce the nutrition-enhanced Artemia that has commercial significance.

Breeding for pollution-safe cultivars (PSCs) can reduce pollutant accumulation in crops. However, the PSC breeding would face the risk of nutritional quality reduction, which is usually ignored in conventional breeding programs targeting to increase crop yield or nutritional quality. Thus, the doubt whether the risk would exist has to be clarified for supporting the PSC breeding. In the present study, a newly bred Cd/Pb-PSC of water spinach (Ipomoea aquatic Forsk.) and its parents (QLQ with low-Cd/Pb accumulation ability and T308 with high yield) of water spinach were employed to clarify the above-mentioned issue. Yields, and concentrations of Cd, Pb, nitrite, and organic and inorganic nutrients in shoots of the three experimental lines were determined. There were no significant differences in Cd/Pb concentration between the new PSC and QLQ, in nitrite content between the new PSC and its two parents and in yield between the new PSC and T308. It is decisively significant that shoot concentrations of organic and inorganic nutrients in the Cd/Pb-PSC were as high as those in one of its parents. It is affirmed that the breeding operations (crossing and consequently continuous selfing) for lowering Cd/Pb accumulation capacity of water spinach would not lower the nutritional values of the obtained Cd/Pb-PSCs from the breeding, which should be a pillar that supports the feasibility to minimize Cd/Pb pollution in vegetables using PSC-breeding method.

The difference in contents of paclitaxel and 10-deacetylbaccatin III (10-DABIII) in needles between wildtype (WT) and a new cultivar (Zhongdayihao, ZD1) of Taxus yunnanensis was examined. Transcriptome profiling was conducted for different tissues of the ZD1 and WT to illustrate the regulation mechanism of paclitaxel biosynthesis. It was observed that average contents of paclitaxel and 10-DABIII in ZD1 were 4 folds and 32 folds higher than those in WT, respectively. More significant elevations of differential expressed genes (DEGs) from paclitaxel biosynthesis pathway were revealed in ZD1 rather than WT, which should be responsible for the higher contents of paclitaxel and 10-DABIII in the ZD1. Special tissues-dependent expression patterns of paclitaxel biosynthesis DEGs in ZD1 compared to WT were unraveled. The relative higher expressions of paclitaxel biosynthesis genes in needles than other tissues supported the higher content of paclitaxel and 10-DABIII content in needles of ZD1. Attenuation of plant hormone signal transduction pathway led to the lower expression of TFs in ZD1 rather than WT. Besides, the significant negative correlations between differential expressed TFs and DEGs from paclitaxel biosynthesis pathway displayed a possibly negative regulation pattern of these TFs on paclitaxel biosynthesis pathway genes. These results provided new insights into the molecular process of paclitaxel synthesis in Taxus.

Screening salt-tolerant crops at different salt concentrations is of great significance but time-consuming. It remains a major challenge to automatically find the appropriate stress concentration to identify the salt tolerance of crops by using the recorded electrical signals of plants over long periods of time. To solve this problem, we designed a data-driven signal dynamics prediction model (SLSTM-TCNN), based on a one-dimensional convolutional neural network (1D-CNN) combined with a long short-term memory neural network. Furthermore, we developed a quantitative model, named the NaCl stress concentration discrimination model (SCDM), to investigate the relationship between the electrical signals, NaCl stress concentration, and time dependence, and used a salt tolerance classification model (STCM) to discover the most appropriate NaCl stress concentration for distinguishing the salt tolerance of wheat. These methods perform the time-consuming task of selecting salt-tolerant varieties of plants under different NaCl concentrations. The results show that the SLSTM-TCNN could quickly predict the signal dynamics of wheat leaves DeKang961 (salt-tolerant) and Langdon (salt-sensitive) under the ongoing stress of different NaCl concentrations. The accuracy of SCDM for differentiating NaCl stress concentrations increased to 88% and 83%, and that of STCM for classifying salt-tolerant and salt-sensitive varieties reached 92.36%. Finally, it was found that the salt tolerance of the two varieties (DeKang961 and Langdon) was higher when the NaCl concentration was in the range of 50–200 mM. In the future, the method will be a potentially useful tool for identifying the salt tolerance of other crops at the seedling stage.

Patients with non-small cell lung cancer (NSCLC) with activating epidermal growth factor receptor (EGFR) mutations (exon 19 deletions and L858R) benefit from EGFR tyrosine kinase inhibitors (TKIs). However, some researchers have reported that responses to TKIs differ by subtypes of EGFR exon 19 mutations. We retrospectively analyzed EGFR exon 19 deletion subtypes and their correlation with clinical outcomes of treatment with TKIs. A cohort of 2664 consecutive patients with NSCLC was enrolled. A total of 440 EGFR exon 19 deletions were defined as 39 subtypes. Among them, 158 patients with advanced lung adenocarcinoma with EGFR exon 19 deletion mutations received EGFR-TKIs. There were no significant differences in progression-free survival or overall survival among patients with non-LRE deletions, delE746, or delL747 (P = 0.463 and P = 0.464, respectively). Furthermore, two patients with EGFR exon19 insertion had durable response to EGFR-TKIs. In conclusion, EGFR exon 19 is highly fragile, resulting in many different deletion and insertion subtypes. There were no significant differences in clinical outcomes after TKI treatment across the different subtypes. It is necessary to attempt to identify all patients with exon 19 deletions so that they can be offered TKI treatment.

As the active site models of [FeFe]-hydrogenases, two new aminophosphine-monosubstituted diiron dithiolate compelxes with the general formulae Fe2(μ-pdt)(CO)5{PPh2NH(C6H4R-p)} (pdt = SCH2CH2CH2S; R = Br (1), Me (2)), were prepared by the oxidative decarbonylation of parent Fe2(μ-pdt)(CO)6 (A) with monophosphines Ph2PNH(C6H4Br-p) or Ph2PNH(C6H4Me-p) in the presence of decarbonylating agent Me3NO·2H2O in MeCN. Interestingly, the ambient temperature treatment of A and aminodiphosphine Ph2PN(C6H4Me-p)PPh2 resulted in the formation of the major chelated complex Fe2(μ-pdt)(CO)4{κ2-Ph2PN(C6H4Me-p)PPh2} (3) concomitant with the minor complex 2. All the complexes 1-3 have been well characterized by elemental analysis, FTIR, NMR (1H, 31P) spectroscopies, and X-ray crystallography. Additionally, the electrochemical and electrocatalytic properties of complexes 1-3 are studied using cyclic voltammetry, where the ratios of the catalytic current (icat) to the reductive peak current (ip) for similar complexes 1, 2, and parent A at 10 mM HOAc exhibit a ranking of 2 ≈ 1 > A.

Zearalenone (ZEA), a kind of nonsteroidal mycotoxin with estrogenic effects, can influence animal reproductive capacity through interfering with estrogen signaling pathway. Previous studies have shown exposure to ZEA at high doses (higher than No-Observed Effect Level, NOEL) had a significant impact on mouse sperm quality and pregnant rate, but little is known about the effect of exposure to ZEA at low doses (lower than NOEL) on mouse reproductive capacity. This study evaluated the effects of exposure to low-dose ZEA on mouse spermatogenesis and semen quality. Male mice (CD-1) of 21days were exposed to ZEA at 20, or 40μg/kg body weight for 14, 28 or 42days. After exposure to ZEA for 14days, the spermatogenic cells in seminiferous tubules were declined dose-independently; however in groups treated by ZEA for 28days, the spermatogenic cells were declined dose-dependently. Moreover, after treatment for 28days or 42days, the DNA double stand break (DSB) in spermatogenic cells were increased in a dose-dependent manner in treated groups. Compared with the control group, the sperm concentration, viability, motility, and hyperactive rate in treated groups were decreased dose-dependently and time-dependently. Meanwhile, deformity and mortality rate of sperm in treated groups were increased remarkably dose-dependently too. In conclusion, low dose ZEA impaired male reproductive capacity especially in spermatogenesis and semen quality of mouse.

Cadmium (Cd) contamination in arable soils presents a serious threat to agricultural crops and public health. Cd pollution-safe cultivars (Cd-PSCs) can effectively restrict human intake of Cd through food chain. However, not much is known on the genetic basis of low-Cd-accumulating trait. Here, we aim to investigate the microRNA (miRNA)-mediated post-transcriptional regulation underlying different capacities of Cd accumulation in Brassica parachinensis L. Using small RNA sequencing, miRNAs were identified and characterized in two previously identified cultivars, a low- (SJ19) and a high-Cd-accumulating cultivar (CX4) of B. parachinensis. 18 known and 148 novel miRNAs were identified to be differentially expressed in response to Cd (Cd-responsive miRNAs) in the two cultivars. Distinct Cd responsive pathways were employed by the two cultivars. In CX4, the overexpression of miR395 was indicated to engage in Cd tolerance by regulating sulfur assimilation and associated with its high Cd accumulation. In SJ19, miRNA-mediated oxidative resistance was enhanced in response to Cd exposure. Furthermore, miR397, miR393 and miR160 were involved in the Cd-induced growth improvement in both roots and shoots of the low-Cd-accumulating cultivar. Our findings provide new insights into miRNAs-mediated Cd response in B. parachinensis, and shed light on molecular-assisted Cd-PSC screening and breeding for agricultural crops and vegetables.

Purpose: Although clinical studies have shown promise for targeting programmed cell death protein-1 (PD-1) and ligand (PD-L1) signaling in non–small cell lung cancer (NSCLC), the factors that predict which subtype patients will be responsive to checkpoint blockade are not fully understood. Experimental Design: We performed an integrated analysis on the multiple-dimensional data types including genomic, transcriptomic, proteomic, and clinical data from cohorts of lung adenocarcinoma public (discovery set) and internal (validation set) database and immunotherapeutic patients. Gene set enrichment analysis (GSEA) was used to determine potentially relevant gene expression signatures between specific subgroups. Results: We observed that TP53 mutation significantly increased expression of immune checkpoints and activated T-effector and interferon-γ signature. More importantly, the TP53/KRAS comutated subgroup manifested exclusive increased expression of PD-L1 and a highest proportion of PD-L1+/CD8A+. Meanwhile, TP53- or KRAS-mutated tumors showed prominently increased mutation burden and specifically enriched in the transversion-high (TH) cohort. Further analysis focused on the potential molecular mechanism revealed that TP53 or KRAS mutation altered a group of genes involved in cell-cycle regulating, DNA replication and damage repair. Finally, immunotherapeutic analysis from public clinical trial and prospective observation in our center were further confirmed that TP53 or KRAS mutation patients, especially those with co-occurring TP53/KRAS mutations, showed remarkable clinical benefit to PD-1 inhibitors. Conclusions: This work provides evidence that TP53 and KRAS mutation in lung adenocarcinoma may be served as a pair of potential predictive factors in guiding anti–PD-1/PD-L1 immunotherapy. Clin Cancer Res; 23(12); 3012–24. ©2016 AACR.

International audience; Images of the nucleus and the coma (gas and dust) of comet 67P/Churyumov– Gerasimenko have been acquired by the OSIRIS (Optical, Spectroscopic and Infrared Remote Imaging System) cameras since 2014 March using both the wide-angle camera and the narrow-angle camera (NAC). We use images from the NAC camera to study a bright outburst observed in the Southern hemisphere on 2015 July 29. The high spatial resolution of the NAC is needed to localize the source point of the outburst on the surface of the nucleus. The heliocentric distance is 1.25 au and the spacecraft–comet distance is 186 km. Aiming to better understand the physics that led to the outgassing, we used the Direct Simulation Monte Carlo method to study the gas flow close to the nucleus and the dust trajectories. The goal is to understand the mechanisms producing the outburst. We reproduce the opening angle of the outburst in the model and constrain the outgassing ratio between the outburst source and the local region. The outburst is in fact a combination of both gas and dust, in which the active surface is approximately 10 times more active than the average rate found in the surrounding areas. We need a number of dust particles 7.83 × 1011 to 6.90 × 1015 (radius 1.97–185 μm), which correspond to a mass of dust (220–21) × 103 kg.

Heavy metals have serious health consequences and ecosystem impacts. A pot experiment was conducted to investigate the variation of cadmium (Cd) uptake and accumulation among 40 cultivars of radish (Raphanus sativus L.) at three Cd levels, including 0.31 (T1), 0.83 (T2), and 1.13 (T3) mg kg−1. Most of the tested cultivars had higher taproot biomass in the T3 treatment when compared to those in the T1 treatment, indicating a Cd stress-induced growth in radish. Taproot Cd concentrations in 95 and 5% of the tested cultivars were lower than 0.1 mg kg−1 (fresh weight, FW) in the T1 and T2 treatments, respectively; however, there was no cultivar suitable for safe consumption in the T3 treatment. Radish production showed potential risk of Cd pollution as high as some leafy vegetables when grown in the soils where Cd concentration exceeded 0.8 mg kg−1. When compared with Chinese heat-resisting or imported cultivars, Chinese common cultivars had significantly higher taproot Cd concentrations. Three low-Cd cultivars and five high-Cd cultivars were identified. Taproot Cd concentrations showed significant correlations between any two of the three treatments (p < 0.01), suggesting that Cd accumulation in taproot of radish was genotype-dependent.

MicroRNAs have been used as diagnostic and prognostic biomarkers for many cancers including oral squamous cell carcinoma (OSCC). Several studies have been shown that microRNA (miRNA) play important roles during the progression of OSCC. However, the results vary largely in different studies due to different platforms and sample sizes. In this study, we systematically evaluated a large scale of miRNA profiles from current qualified OSCC samples, and further investigated the functions of genes regulated by these key miRNAs as well as the signaling pathways through which these miRNA effect carcinogenesis. Seven key miRNAs were identified, and of which three were significantly upregulated, including hsa-miR-21, hsa-miR-31, hsa-miR-338, and four were downregulated, namely hsa-miR-125b, hsa-miR-133a, hsa-miR-133b, and hsa-miR-139. The function enrichment analysis revealed that target genes of upregulated miRNAs were associated with cellular protein metabolic process, macromolecule metabolic process, and TGF-beta pathway, while the targets of downregulated were enriched in negative regulation of macromolecule biosynthetic process and gene expression, and p53, long-term potentiation and adherens junction pathways. Transcription factor analysis revealed that there were 67 (51.1%) transcription factors influenced by both up and downregulated miRNAs. In summary, seven key miRNAs were found to play essential role in progression of OSCC, as well as the target genes and transcription factors of these miRNAs. The potential functions of these target genes identified in our study may be profitable to diagnosis and prognostic prediction of OSCC as biomarkers. J. Cell. Physiol. 232: 2178-2185, 2017. © 2016 Wiley Periodicals, Inc.

Metabolic reprogramming has become a hot topic recently in the regulation of tumour biology. Although hundreds of altered metabolic genes have been reported to be associated with tumour development and progression, the important prognostic role of these metabolic genes remains unknown. We downloaded messenger RNA expression profiles and clinicopathological data from The Cancer Genome Atlas and the Gene Expression Omnibus database to uncover the prognostic role of these metabolic genes. Univariate Cox regression analysis and lasso Cox regression model were utilized in this study to screen prognostic associated metabolic genes. Patients with high-risk demonstrated significantly poorer survival outcomes than patients with low-risk in the TCGA database. Also, patients with high-risk still showed significantly poorer survival outcomes than patients with low-risk in the GEO database. What is more, gene set enrichment analyses were performed in this study to uncover significantly enriched GO terms and pathways in order to help identify potential underlying mechanisms. Our study identified some survival-related metabolic genes for rectal cancer prognosis prediction. These genes might play essential roles in the regulation of metabolic microenvironment and in providing significant potential biomarkers in metabolic treatment.

Cultivar-dependent cadmium (Cd) accumulation was principal in developing Cd-pollution safe cultivars (PSCs). Proteins related to different Cd accumulations of the low-Cd-accumulating (SJ19) and high-Cd-accumulating (CX4) cultivars were investigated by iTRAQ analysis. Higher Cd bioaccumulation factors and translocation factor in CX4 than in SJ19 were consistent with the cultivar-dependent Cd accumulations. The Cd uptake was promoted in CX4 due to its higher expression of Cd-binding proteins and the lower expression of Cd-efflux proteins in roots. What's more, significantly elevated thiol groups (PC2 and PC3) in CX4 under Cd stress might contribute to the high Cd accumulation in roots and the root-to-shoot translocation of Cd-PC complex. Up-regulated proteins involved in cellulose biosynthesis and pectin de-esterification in SJ19 enhanced the Cd sequestration of root cell walls, which was considered as the predominant strategy for reducing Cd accumulation in shoots. The present study provided novel insights in the cultivar-dependent Cd accumulation in shoots of B. parachinensis.

Ovarian cancer (OC) is the highest frequent malignant gynecologic tumor with very complicated pathogenesis. The purpose of the present academic work was to identify significant genes with poor outcome and their underlying mechanisms. Gene expression profiles of GSE36668, GSE14407 and GSE18520 were available from GEO database. There are 69 OC tissues and 26 normal tissues in the three profile datasets. Differentially expressed genes (DEGs) between OC tissues and normal ovarian (OV) tissues were picked out by GEO2R tool and Venn diagram software. Next, we made use of the Database for Annotation, Visualization and Integrated Discovery (DAVID) to analyze Kyoto Encyclopedia of Gene and Genome (KEGG) pathway and gene ontology (GO). Then protein-protein interaction (PPI) of these DEGs was visualized by Cytoscape with Search Tool for the Retrieval of Interacting Genes (STRING). There were total of 216 consistently expressed genes in the three datasets, including 110 up-regulated genes enriched in cell division, sister chromatid cohesion, mitotic nuclear division, regulation of cell cycle, protein localization to kinetochore, cell proliferation and Cell cycle, progesterone-mediated oocyte maturation and p53 signaling pathway, while 106 down-regulated genes enriched in palate development, blood coagulation, positive regulation of transcription from RNA polymerase II promoter, axonogenesis, receptor internalization, negative regulation of transcription from RNA polymerase II promoter and no significant signaling pathways. Of PPI network analyzed by Molecular Complex Detection (MCODE) plug-in, all 33 up-regulated genes were selected. Furthermore, for the analysis of overall survival among those genes, Kaplan–Meier analysis was implemented and 20 of 33 genes had a significantly worse prognosis. For validation in Gene Expression Profiling Interactive Analysis (GEPIA), 15 of 20 genes were discovered highly expressed in OC tissues compared to normal OV tissues. Furthermore, four genes (BUB1B, BUB1, TTK and CCNB1) were found to significantly enrich in the cell cycle pathway via re-analysis of DAVID. In conclusion, we have identified four significant up-regulated DEGs with poor prognosis in OC on the basis of integrated bioinformatical methods, which could be potential therapeutic targets for OC patients.

Deficiency of surfactant proteins (SPs) is the main cause of respiratory distress syndrome (RDS) and chronic lung diseases. Our previous study demonstrated that miR‑431 was differentially expressed between infants with RDS and infants without RDS using microarray analysis. However, the potential role of miR‑431 in the development of lung function is still unknown. In the present study, the morphological characteristics of lung tissues and the expression levels of miR‑431 were examined at three time points of rat lung development [gestational days 19 and 21 (E19, and E21) and postnatal day (P3)]. The protein and mRNA levels of SMAD4 and SPs (SP‑A, SP‑B, SP‑C and SP‑D) were also validated by reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) and western blot analysis, respectively. The expression levels of miR‑431 were gradually decreased over time periods of E19, E21 and P3, as determine using RT‑qPCR and fluorescence in situ hybridization. Dual luciferase‑reporter assays revealed that SMAD4 is a direct target of miR‑431. The mRNA and protein expression levels of SMAD4 and SPs increased gradually in rat lung tissues from E19 to P3. The order of magnitude was as follows: E19, E21 and P3. The present study demonstrated that the expression level of miR‑431 decreased in the order of E19, E21 and P3 during rat lung development. The target gene of miR‑431, SMAD4, was negatively regulated by miR‑431, and its expression levels in the rat lung tissue increased from E19 to the P3. Surfactant synthesis was further increased over the E19 to P3 time period. Further studies are required to determine how miR‑431 regulates pulmonary surfactant synthesis by targeting SMAD4.

Flowering, the beginning of the reproductive growth, is a significant stage in the growth and development of plants. Conifers are economically and ecologically important, characterized by straight trunks and a good wood quality and, thus, conifer plantations are widely distributed around the world. In addition, conifer species have a good tolerance to biotic and abiotic stress, and a stronger survival ability. Seeds of some conifer species, such as Pinus koraiensis, are rich in vitamins, amino acids, mineral elements and other nutrients, which are used for food and medicine. Although conifers are the largest (giant sequoia) and oldest living plants (bristlecone pine), their growth cycle is relatively long, and the seed yield is unstable. In the present work, we reviewed selected literature and provide a comprehensive overview on the most influential factors and on the methods and techniques that can be adopted in order to improve flowering and seed production in conifers species. The review revealed that flowering and seed yields in conifers are affected by a variety of factors, such as pollen, temperature, light, water availability, nutrients, etc., and a number of management techniques, including topping off, pruning, fertilization, hormone treatment, supplementary pollination, etc. has been developed for improving cone yields. Furthermore, several flowering-related genes (FT, Flowering locus T and MADS-box, MCMI, AGAMOUS, DEFICIENCES and SRF) that play a crucial role in flowering in coniferous trees were identified. The results of this study can be useful for forest managers and for enhancing seed yields in conifer plantations for commercial use.

Cadmium (Cd) is a highly toxic heavy metal and has spread widely in the environment in recent decades. This review summarizes current knowledge about Cd contamination of leafy vegetables, its toxicity, exposure, health risks, and approaches to reducing its toxicity in humans. Leafy vegetable consumption has been identified as a dominant exposure pathway of Cd in the human body. An overview of Cd pollution in leafy vegetables as well as the main sources of Cd is given. Notable estimated daily intakes and health risks of Cd exposure through vegetable consumption for humans are revealed in occupational exposure areas and even in some reference areas. Vegetable consumption is one of the most significant sources of exposure to Cd, particularly in occupational exposure regions. Therefore, numerous approaches have been developed to minimize the accumulation of Cd in leafy vegetables, among which the breeding of Cd pollution-safe cultivars is one of the most effective tools. Furthermore, dietary supplements from leafy vegetables perform positive roles in alleviating Cd toxicity in humans with regard to the effects of essential mineral elements, vitamins and phytochemicals taken into the human body via leafy vegetable consumption.

Sialic acid modification is a kind of post-translational modification. To investigate the regulation effect of sialic acid on neural differentiation, we used CycloManN propanyl perac (CycloManN pro), a metabolic precursor of sialic acid, to treat PC12 cells. We noted that CycloManN pro indeed robustly promoted global sialylation detected by MAL II lectin blot in PC12 cells. Simultaneously, we interestingly found that the neurite outgrowth of PC12 cells was significantly promoted by the CycloManN pro treatment. The profile analysis of sialylated proteins showed that a protein band at 55KD was greatly enhanced especially in PC12L cells after CycloManN pro treatment. After enrichment with lectin MAL II, the proteins in this band were analyzed by mass spectrometry. The results showed that 23 proteins were in the band, but the score of vimentin was the highest among them. To investigate further the role of vimentin in the process of neurite differentiation, vimentin construct was transfected into PC12 cells. We interestingly observed that ectopic expression of vimentin significantly enhanced the neurite outgrowth induced by CycloManN pro. However, after three potential glycosylation sites (Ser-7, Thr-33, Ser-34:) of vimentin were mutated to alanine, overexpression of the mutated vimentin completely lost the enhancement activity for the neural differentiation even in the presence of CycloManN pro. Taken together, our study demonstrated that vimentin was important in the induction of neural differentiation by CycloManN pro.

The phenomenon of delayed flowering after the application of nitrogen (N) fertilizer has long been known in agriculture, but the detailed molecular basis for this phenomenon is largely unclear. Here we used a modified method of suppression-subtractive hybridization to identify two key factors involved in N-regulated flowering time control in Arabidopsis thaliana, namely ferredoxin-NADP(+)-oxidoreductase and the blue-light receptor cryptochrome 1 (CRY1). The expression of both genes is induced by low N levels, and their loss-of-function mutants are insensitive to altered N concentration. Low-N conditions increase both NADPH/NADP(+) and ATP/AMP ratios, which in turn affect adenosine monophosphate-activated protein kinase (AMPK) activity. Moreover, our results show that the AMPK activity and nuclear localization are rhythmic and inversely correlated with nuclear CRY1 protein abundance. Low-N conditions increase but high-N conditions decrease the expression of several key components of the central oscillator (e.g., CCA1, LHY, and TOC1) and the flowering output genes (e.g., GI and CO). Taken together, our results suggest that N signaling functions as a modulator of nuclear CRY1 protein abundance, as well as the input signal for the central circadian clock to interfere with the normal flowering process.