G Protein Subunit ß1 Facilitates Influenza A Virus Replication by Promoting the Nuclear Import of PB2.

G protein subunit ß1 (GNB1), the beta subunit of the G protein family, plays an important role in regulating transmembrane signal transduction. Although a recent study has demonstrated that GNB1 can bind the matrix protein 1 (M1) to facilitate M1 transport to budding sites and promote the release of progeny influenza A virus (IAV), whether the GNB1 protein has other functions in IAV replication requires further study. Here, we found that GNB1 promoted IAV replication, as virus yield decreased in GNB1 knockdown or knockout cells. GNB1 interacted with polymerase subunits PB2, PB1, and PA. Overexpressed GNB1 facilitated PB2 binding to importin α3, α5, and α7 promoting the nuclear import of PB2, enhancing viral RNA synthesis and polymerase activity. Altogether, our results demonstrated that GNB1 positively regulates virus replication by interacting with polymerase subunits and facilitating the nuclear import of PB2, which provide novel insights into the molecular mechanism of IAV. IMPORTANCE Until now, there has been only one article on the role of GNB1 in IAV budding. No study has investigated the role of GNB1 in IAV replication. In this study, our research demonstrated that GNB1 could increase the interaction between PB2 and the importin α isoform and mediate the nuclear import of PB2. Therefore, GNB1 could promote viral replication and transcription. Our results provide a better understanding of the molecular mechanisms of viral replication and provide potential antiviral drug targets.

Comparative multi-omics analysis of hypoxic germination tolerance in weedy rice embryos and coleoptiles.

Hypoxic germination tolerance is an important trait for seedling establishment of direct-seeded rice. Our comparative metabolomics analysis revealed that weedy rice accumulated more sugar and amino acids than cultivated rice accumulated in the embryo and coleoptile tissues under hypoxic stress. At the transcriptional level, oxidative phosphorylation activity in weedy rice was higher than in cultivated rice that likely led to more efficient energy metabolism during hypoxic stress. Based on our comparative proteomics analysis, enriched proteins related to cell wall implied that the advantages in energy metabolism of weedy rice were ultimately reflected in the formation of tissue structures. In this study, we found that most of key hypoxic germination tolerance (HGT) genes shared the same genetic backgrounds with Oryza japonica, however, several of them genetically similar to other Oryza plant also play important roles. Our findings suggest weedy rice can serve as genetic resources for the improvement of direct-seeding rice.

Retrospective analysis of the efficacy and influencing factors of autologous hematopoietic stem cell transplantation for multiple myeloma.

This study aims to review the clinical efficacy and factors affecting the treatment of multiple myeloma (MM) by autologous hematopoietic stem cell transplantation (ASCT). The clinical data of 47 patients with MM from the Department of Hematology of Henan Cancer Hospital from September 2010 to July 2018 were retrospectively analyzed. At pre-transplantation of autologous cells, 25.5% were in complete remission (CR), 14.9% were in very good partial remission (VGPR) and 59.6% were in partial remission (PR). Among these cases, one case had PR after three recurrences. At post-transplantation, 51% were in CR, including two cases who received double transplantations, 27.7% were in VGPR, and 21.3% were in PR. The median follow-up time was 27.6 months (4-96 months). The 3-year progression free survival (PFS) and overall survival (OS) were 47.9% and 79.6%, respectively. The Analysis of variance (ANOVA) results revealed that factors that affected OS were international staging system (ISS) stage (P = 0.002), CR and VGPR post-transplantation (P = 0.002), while factors that affected PFS were ISS stage (P = 0.005), pre-transplant induction therapy (P = 0.032), and disease risk stratification (P = 0.017). The curative effects for PFS were CR and VGPR pre-transplantation (P = 0.013) and post-transplantation (P = 0.011). The Cox multivariate regression analysis revealed that ISS stage and CR and VGPR post-transplantation were independent prognostic factors of OS. At post-transplantation, CR and VGPR, ISS stage, and pre-transplant induction therapy were independent prognostic factors for PFS. In conclusion, ASCT can improve the clinical efficacy and survival rate of MM patients. ISS stage, CR and VGPR post-transplantation are independent prognostic factors of OS and PFS, while pre-transplant induction therapy is an independent prognostic factor for PFS.

Development of sandwich Enzyme-Linked Immunosorbent Assay for the detection of porcine epidemic diarrhea virus in fecal samples.

Porcine epidemic diarrhea virus (PEDV) causes acute diarrhea and dehydration in new-born piglets with subsequent economic losses to swine industry. In the current study, gene encoding of 381aa-792aa spike protein (S1) with the main epitope relative to virus neutralization of PEDV was amplified by RT-PCR and inserted into vector pET-30A(+). The plasmid was transferred into Escherichia coli BL21 (DE3). Meanwhile, recombinant protein expression was induced by isopropy1-ß-galactopyranoside (IPTG). After denaturation and renaturation of inclusion bodies, the S1 protein was obtained by using purified recombinant S1 protein in immunized female BALB/c mice. Monoclonal antibodies (MAb) against S1 protein, named 4C7 by hybridoma technique were gained successfully. The result showed that MAb can specifically respond to S1 protein and PEDV via ELISA, Western bolt and immunofluorescence assay methods. A sandwich ELISA (S-ELISA) was established by using the captured monoclonal antibodies 4C7. The sensitivity and specificity were compared between S-ELISA and RT-PCR, which showed similar sensitivity and specificity. This work indicated that S-ELISA would be a significant tool alongside a specific diagnostic reagent for PEDV in future.

Isoprenylated phenolic compounds with tyrosinase inhibition from Morus nigra.

A new isoprenylated sanggenon-type flavanone, nigrasin K (1), together with three known analogs (2-4) and five known Diels-Alder adducts (5-9), were isolated from the twigs of Morus nigra. Their structures were elucidated by spectroscopic methods. Sanggenon M (2), chalcomoracin (5), sorocein H (6), kuwanon J (7), sanggenon C (8), and sanggenon O (9) showed significant inhibitory effects on mushroom tyrosinase.

Seroprevalence investigation of bovine ephemeral fever in yaks in Tibetan Plateau of China from 2012 to 2015.

Bovine ephemeral fever (BEF) is caused by the arthropod-borne bovine ephemeral fever virus (BEFV), which is classified in family Rhabdoviridae and genus Ephemerovirus. However, it is still unclear whether yaks from the Tibetan plateau of China are exposed to BEFV. It is the first time that a survey was conducted to investigate the seroprevalence of BEFV infection in yaks (Bos grunniens) on the Tibetan Plateau of China. A total of 1123 serum samples were collected randomly from yaks from 2012 to 2015 and were assayed for BEFV antibodies by enzyme-linked immunosorbent assay (ELISA). The proportions of positive serum samples were assessed among the 1123 samples, as well as factors of geographical origin and years. The results showed that there were 454 serum samples that tested positive for BEFV, and the total positive rate is 40.4 %. The prevalence in 2012, 2013, 2014, and 2015 was 49.3, 36, 44.1, and 34.0 %, respectively, and the difference is statistically significant (P< 0.01). In different regions, the prevalence was ranged from 34.7 to 45.7 % with a significant difference among the different regions of (P < 0.05). Logistic regression analysis showed that yaks in Tibet (Xizang autonomous region) (45.7 %) had 1.6 times (OR = 1.589, 95 % CI = 1.141-2.215, P < 0.01) higher risk of being seropositive compared to yaks in Qinghai province, while no regional difference was found of Sichuan province compared to Qinghai (P > 0.05). The prevalence in 2012 (49.3 %) was more than 1.8 time (OR = 1.880, 95 % CI = 1.350-2.619, P < 0.001) at risk of acquiring the infection compared to the year of 2015. The prevalence of yaks in 2014 (44.1 %) had a 1.5 times (OR = 1.528, 95 % CI = 1.350-2.619, p < 0.001) at risk of being seropositive compared to yaks in 2015, while no year difference was found of 2013 compared to 2015 (P > 0.05). Our study suggests that the yaks from the high plateau are highly infected by BEFV, and geographical origin and years are main risk factors for BEF seroprevalence.

Prevalence, Associated Risk Factors, and Phylogenetic Analysis of Toxocara vitulorum Infection in Yaks on the Qinghai Tibetan Plateau, China.

Toxocara vitulorum has been rarely reported in yaks at high altitudes and remote areas of Sichuan Province of Tibetan Plateau of China. The current study was designed to investigate the prevalence, associated risk factors, and phylogenetic characteristics of T. vitulorum in yak calves on the Qinghai Tibetan plateau. Fecal samples were collected from 891 yak calves and were examined for the presence of T. vitulorum eggs by the McMaster technique. A multivariable logistic regression model was employed to explore variables potentially associated with exposure to T. vitulorum infection. T. vitulorum specimens were collected from the feces of yaks in Hongyuan of Sichuan Province, China. DNA was extracted from ascaris. After PCR amplification, the sequencing of ND1 gene was carried out and phylogenetic analyses was performed by MEGA 6.0 software. The results showed that 64 (20.1%; 95% CI 15.8-24.9%), 75 (17.2; 13.8-21.1), 29 (40.9; 29.3-53.2), and 5 (7.6; 2.5-16.8) yak calves were detected out to excrete T. vitulorum eggs in yak calve feces in Qinghai, Tibet, Sichuan, and Gansu, respectively. The present study revealed that high infection and mortality by T. vitulorum is wildly spread on the Qinghai Tibetan plateau, China by fecal examination. Geographical origin, ages, and fecal consistencies are the risk factors associated with T. vitulorum prevalence by logistic regression analysis. Molecular detection and phylogenetic analysis of ND1 gene of T. vitulorum indicated that T. vitulorum in the yak calves on the Qinghai Tibetan plateau are homologous to preveiously studies reported.

Inhibitory Effects of (2'R)-2',3'-dihydro-2'-(1-hydroxy-1-methylethyl)-2,6'-bibenzofuran-6,4'-diol on Mushroom Tyrosinase and Melanogenesis in B16-F10 Melanoma Cells.

(2'R)-2',3'-Dihydro-2'-(1-hydroxy-1-methylethyl)-2,6'-bibenzofuran-6,4'-diol (DHMB) is a natural compound extracted from Morus notabilis. It was found that DHMB acts as a competitive inhibitor against mushroom tyrosinase with a Ki value of 14.77 µM. Docking results further indicated that it could form strong interactions with one copper ion with a distance of 2.7 Å, suggesting the mechanism of inhibition might be due to chelating copper ions in the active site. Furthermore, melanin production in B16-F10 murine melanoma cells was significantly inhibited by DHMB in a concentration-dependent manner without cytotoxicity. The results of western blotting also showed that DHMB decreased 3-isobuty-1-methxlzanthine-induced mature tyrosinase expression. Taken together, these findings indicated that DHMB may be a new promising pigmentation-altering agent for agriculture, cosmetic, and therapeutic applications.

Mutual antagonism of mouse-adaptation mutations in HA and PA proteins on H9N2 virus replication.

Avian H9N2 viruses have wide host range among the influenza A viruses. However, knowledge of H9N2 mammalian adaptation is limited. To explore the molecular basis of the adaptation to mammals, we performed serial lung passaging of the H9N2 strain A/chicken/Hunan/8.27 YYGK3W3-OC/2018 (3W3) in mice and identified six mutations in the hemagglutinin (HA) and polymerase acidic (PA) proteins. Mutations L226Q, T511I, and A528V of HA were responsible for enhanced pathogenicity and viral replication in mice; notably, HA-L226Q was the key determinant. Mutations T97I, I545V, and S594G of PA contributed to enhanced polymerase activity in mammalian cells and increased viral replication levels in vitro and in vivo. PA-T97I increased viral polymerase activity by accelerating the viral polymerase complex assembly. Our findings revealed that the viral replication was affected by the presence of PA-97I and/or PA-545V in combination with a triple-point HA mutation. Furthermore, the double- and triple-point PA mutations demonstrated antagonistic effect on viral replication when combined with HA-226Q. Notably, any combination of PA mutations, along with double-point HA mutations, resulted in antagonistic effect on viral replication. We also observed antagonism in viral replication between PA-545V and PA-97I, as well as between HA-528V and PA-545V. Our findings demonstrated that several antagonistic mutations in HA and PA proteins affect viral replication, which may contribute to the H9N2 virus adaptation to mice and mammalian cells. These findings can potentially contribute to the monitoring of H9N2 field strains for assessing their potential risk in mammals.

Benzbromarone, an old uricosuric drug, inhibits human fatty acid binding protein 4 in vitro and lowers the blood glucose level in db/db mice.

AIM: Fatty acid-binding protein 4 (FABP4) plays an important role in maintaining glucose and lipid homeostasis. The aim of this study was to find new inhibitors of FABP4 for the treatment of type 2 diabetes. METHODS: Human FABP4 protein was expressed, and its inhibitors were detected in 1,8-ANS displacement assay. The effect of the inhibitor on lipolysis activity was examined in mouse 3T3-L1 preadipocytes. The db/db mice were used to evaluate the anti-diabetic activity of the inhibitor. Molecular docking and site-directed mutagenesis studies were carried out to explore the binding mode between the inhibitor and FABP4. RESULTS: From 232 compounds tested, benzbromarone (BBR), an old uricosuric drug, was discovered to be the best inhibitor of FABP4 with an IC50 value of 14.8 µmol/L. Furthermore, BBR (25 µmol/L) significantly inhibited forskolin-stimulated lipolysis in 3T3-L1 cells. Oral administration of BBR (25 or 50 mg/kg, for 4 weeks) dose-dependently reduced the blood glucose level and improved glucose tolerance and insulin resistance in db/db mice. Molecular docking revealed that the residues Ser55, Asp76, and Arg126 of FABP4 formed important interactions with BBR, which was confirmed by site-directed mutagenesis studies. CONCLUSION: BBR is an inhibitor of FABP4 and a potential drug candidate for the treatment of type 2 diabetes and atherosclerosis.

Discussion on the ecological theory and technological approaches of ecosystem quality improvement and stability enhancement.

Improving ecosystem quality and stability is one of the urgent tasks of national ecological environment construction. However, the ecological theory of ecosystem quality and stability has not been well clarified. Based on the summary of influencing factors and interaction between ecosystem quality and stability, we discussed the ecolo-gical theory on the evolution of ecosystem quality and stability from the perspectives of self-organization of biological agglomeration and structure nesting, correlation of ecological elements and coupling of ecological processes, ecosystem integrity and function emergence, ecological service spillover and efficiency tradeoff, synergy and interactions between resource supply capacity and environmental suitability, as well as interactions between spontaneous change and human activities. Technologies approaches and management strategies were proposed from the aspects of ecosystem macro-pattern adjustment, protected natural areas system construction, regional complex ecosystem comprehensive management, degraded ecosystem restoration, damaged ecosystem reestablishment, typical ecosystem process management.

Mapping Chinese annual gross primary productivity with eddy covariance measurements and machine learning.

Annual gross primary productivity (AGPP) is the basis for grain production and terrestrial carbon sequestration. Mapping regional AGPP from site measurements provides methodological support for analysing AGPP spatiotemporal variations thereby ensures regional food security and mitigates climate change. Based on 641 site-year eddy covariance measuring AGPP from China, we built an AGPP mapping scheme based on its formation and selected the optimal mapping way, which was conducted through analysing the predicting performances of divergent mapping tools, variable combinations, and mapping approaches in predicting observed AGPP variations. The reasonability of the selected optimal scheme was confirmed by assessing the consistency between its generating AGPP and previous products in spatiotemporal variations and total amount. Random forest regression tree explained 85 % of observed AGPP variations, outperforming other machine learning algorithms and classical statistical methods. Variable combinations containing climate, soil, and biological factors showed superior performance to other variable combinations. Mapping AGPP through predicting AGPP per leaf area (PAGPP) explained 86 % of AGPP variations, which was superior to other approaches. The optimal scheme was thus using a random forest regression tree, combining climate, soil, and biological variables, and predicting PAGPP. The optimal scheme generating AGPP of Chinese terrestrial ecosystems decreased from southeast to northwest, which was highly consistent with previous products. The interannual trend and interannual variation of our generating AGPP showed a decreasing trend from east to west and from southeast to northwest, respectively, which was consistent with data-oriented products. The mean total amount of generated AGPP was 7.03 ± 0.45 PgC yr-1 falling into the range of previous works. Considering the consistency between the generated AGPP and previous products, our optimal mapping way was suitable for mapping AGPP from site measurements. Our results provided a methodological support for mapping regional AGPP and other fluxes.

Dew water isotopic ratios and their relationships to ecosystem water pools and fluxes in a cropland and a grassland in China.

Dew formation has the potential to modulate the spatial and temporal variations of isotopic contents of atmospheric water vapor, oxygen and carbon dioxide. The goal of this paper is to improve our understanding of the isotopic interactions between dew water and ecosystem water pools and fluxes through two field experiments in a wheat/maize cropland and in a short steppe grassland in China. Measurements were made during 94 dew events of the D and (18)O compositions of dew, atmospheric vapor, leaf, xylem and soil water, and the whole ecosystem water flux. Our results demonstrate that the equilibrium fractionation played a dominant role over the kinetic fractionation in controlling the dew water isotopic compositions. A significant correlation between the isotopic compositions of leaf water and dew water suggests a large role of top-down exchange with atmospheric vapor controlling the leaf water turnover at night. According to the isotopic labeling, dew water consisted of a downward flux of water vapor from above the canopy (98%) and upward fluxes originated from soil evaporation and transpiration of the leaves in the lower canopy (2%).

New isoprenylated flavones and stilbene derivative from Artocarpus hypargyreus.

Three new isoprenylated flavones, hypargyflavones A-C (1-3, resp.), and one novel stilbene derivative, hypargystilbene A (4), together with seven known compounds, 5-11, were isolated from the stems of Artocarpus hypargyreus Hance. The structures were elucidated by spectroscopic methods. Hypargyflavone A (1), cudraflavone C (8), brosimone I (10), and norartocarpin (11) showed inhibitory effects on pancreatic lipase.

2-Arylbenzofuran and tyrosinase inhibitory constituents of Morus notabilis.

Phytochemical investigation of the stem of Morus notabilis led to the isolation and characterization of 10 compounds of 2-arylbenzofurans (1-10), including two new compounds, (2'R)-2',3'-dihydro-2'-(1-hydroxy-1-methylethyl)-2,6'-bibenzofuran-6,4'-diol (1) and 5,6-dimethoxy-2-(3-hydroxy-5-methoxyphenyl)benzofuran (2). Moracins O (6) and P (10) showed inhibitory effects on mushroom tyrosinase with IC50 values being lower than that of kojic acid.

[Progress of anti-tumor study based on BRAF].

BRAF is one of the most important pro-oncogenes, which is mutated in approximately 8% of human tumors. The most common BRAF mutation is a valine-to-glutamate transition (V600E) that is expressed primarily in melanoma, colorectal cancer and thyroid carcinoma. MEK/ERK is constitutively activated in the cells expressing BRAFV600E, leading to tumor development, invasion, and metastasis. Therefore, BRAFV600E is a therapeutic target for melanoma and some other BRAFV600E tumors. Vemurafenib, a BRAFV600E inhibitor, which was approved by FDA for the treatment of late-stage melanoma in 2011, produces improved rates of overall and progression-free survival in patients with the BRAFV600E mutation, making a dramatic breakthrough in melanoma treatment. Vemurafenib is also an individual target drug based on genetic diagnosis. However, its therapeutic success is limited by the emergence of drug resistance. Therefore, it is important to explore the mechanisms underlying the resistance for developing new inhibitor drugs and for preventing or delaying the resistance evolution to BRAF inhibitor drugs. In this review, we described the role of BRAFV600E as an anti-tumor drug target and the development of BRAF inhibitors. We also discussed the mechanisms leading to resistance of BRAFV600E inhibitors. Furthermore, therapeutic strategies that might be employed to overcome acquired resistance were proposed.

[Ecological economics foundation research on ecological values, ecological asset management, and value realization: Scientific concepts, basic theories, and realization paths]. / è‡ªç„¶ç”Ÿæ€ä»·å€¼ã€ç”Ÿæ€èµ„äº§ç®¡ç†åŠä»·å€¼å®žçŽ°çš„ç”Ÿæ€ç»æµŽå­¦åŸºç¡€ç ”ç©¶­­ç§‘学概念、基础理论及实现途径.

Ecosystems provide various environmental conditions and natural resources for life, production, and human livelihoods. Ecosystem management based on natural laws is the basic way for human society to recognize the value of nature and ecosystems, protect and utilize the natural environment and resources, create and accumulate ecological assets, and sustainably develop. Taking that as one of the core concepts, regional ecological economics or economic ecology is becoming a frontier in scientific research to evaluate the Anthropocene earth system and sustainable development. From the perspective of macro-ecosystem science, based on the multi-functionality and multi-values of ecosystems, this study expounded the concepts of natural ecological value, ecological assets, ecological products, and some related phrases, analyzed the processes and principles of formation and change of ecological assets, production and consumption of ecological products, ecological investment, and profit and loss of ecological assets based on integrated ecology, sociology, and economics, proposed paths of regional ecological asset management and ecological value realization. It would provide theoretical and methodological references for the evaluation of regional ecosystem value and ecological assets, and the development of ecological industries and ecological value realization systems.

Integrated analysis of the gut microbiome and metabolome in a mouse model of inflammation-induced colorectal tumors.

Colorectal cancer (CRC) is a common malignancy worldwide, and the gut microbiota and metabolites play an important role in its initiation and progression. In this study, we constructed a mouse model of inflammation-induced colorectal tumors, with fixed doses of azoxymethane/dextran sulfate sodium (AOM/DSS). We found that colorectal tumors only formed in some mice treated with certain concentrations of AOM/DSS (tumor group), whereas other mice did not develop tumors (non-tumor group). 16S rDNA amplicon sequencing and liquid chromatography-mass spectrometry (LC-MS)/MS analyses were performed to investigate the microbes and metabolites in the fecal samples. As a result, 1189 operational taxonomic units (OTUs) were obtained from the fecal samples, and the non-tumor group had a relatively higher OTU richness and diversity. Moreover, 53 different microbes were identified at the phylum and genus levels, including Proteobacteria, Cyanobacteria, and Prevotella. Furthermore, four bacterial taxa were obviously enriched in the non-tumor group, according to linear discriminant analysis scores (log10) > 4. The untargeted metabolomics analysis revealed significant differences between the fecal samples and metabolic phenotypes. Further, the heatmaps and volcano plots revealed 53 and 19 dysregulated metabolites between the groups, in positive and negative ion modes, respectively. Styrene degradation and amino sugar-nucleotide sugar metabolism pathways were significantly different in positive and negative ion modes, respectively. Moreover, a correlation analysis between the metabolome and microbiome was further conducted, which revealed the key microbiota and metabolites. In conclusion, we successfully established a tumor model using a certain dose of AOM/DSS and identified the differential intestinal microbiota and characteristic metabolites that might modulate tumorigenesis, thereby providing new concepts for the prevention and treatment of CRC.

[Discussion on the ecological theory and assessment methods of ecosystem quality and its evolution]. / ç”Ÿæ€ç³»ç»Ÿè´¨é‡åŠå ¶çŠ¶æ€æ¼”å˜çš„ç”Ÿæ€å­¦ç†è®ºå’Œè¯„ä¼°æ–¹æ³•ä¹‹æŽ¢ç´¢.

Ecological civilization construction and ecological environment governance are basic tasks of state gover-nance in China. China has clearly put forward the goal of improving ecosystem quality and stability. However, there are no consensus on the scientific concept of ecosystem quality and the assessment methods of ecosystem quality evolution, which has puzzled the academic community. Based on the summarization of the scientific concept of ecosystem quality and its evolution, we discussed the concept of ecosystem quality and its ecological theory basis by referencing the concept of production quality, quality management and quality assessment of material production. The scientific connotation of ecosystem quality and its evolution was discussed from the perspectives of ecosystem natural attribute-social attribute-economic attribute and the relationships between them, the cascade relations of ecosystem component-structure-process-function-service-efficacy, the feedback of factor-system-environment, and the logic relation of state ecosystem fluctuation-quantity variation-quality alternation. We proposed perspectives and approaches of multi-objective assessment of ecosystem quality alternation from the aspects of natural resource environment system, typical ecosystem, regional macro-ecosystem, and ecological engineering efficacy.