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Allelopathy means that one plant produces chemical substances to affect the growth and development of other plants. Usually, allelochemicals can stimulate or inhibit the germination and growth of plants, which have been considered as potential strategy for drug development of environmentally friendly biological herbicides. Obviously, the discovery of plant materials with extensive sources, low cost and markedly allelopathic effect will have far-reaching ecological impacts as the biological herbicide. At present, a large number of researches have already reported that certain plant-derived allelochemicals can inhibit weed growth. In this study, the allelopathic effect of Artemisia argyi was investigated via a series of laboratory experiments and field trial. Firstly, water-soluble extracts exhibited the strongest allelopathic inhibitory effects on various plants under incubator conditions, after the different extracts authenticated by UPLC-Q-TOF-MS. Then, the allelopathic effect of the A. argyi was systematacially evaluated on the seed germination and growth of Brassica pekinensis, Lactuca sativa, Oryza sativa, Portulaca oleracea, Oxalis corniculata and Setaria viridis in pot experiments, it suggested that the A. argyi could inhibit both dicotyledons and monocotyledons not only by seed germination but also by seedling growth. Furthermore, field trial showed that the A. argyi significantly inhibited the growth of weeds in Chrysanthemum morifolium field with no adverse effect on the growth of C. morifolium. At last, RNA-Seq analysis and key gene detection analysis indicated that A.argyi inhibited the germination and growth of weed via multi-targets and multi-paths while the inhibiting of chlorophyll synthesis of target plants was one of the key mechanisms. In summary, the A. argyi was confirmed as a potential raw material for the development of preventive herbicides against various weeds in this research. Importantly, this discovery maybe provide scientific evidence for the research and development of environmentally friendly herbicides in the future.

The feasibility and efficiency of magnetite nanoparticles (Fe3O4NPs) enhanced bio-hydrogen production from glucose anaerobic fermentation were evaluated in this study. The results demonstrated that the maximum hydrogen yield (HY) of 12.97 mL H2/g-VSS was obtained with 50 mg/L and 40–60 nm of Fe3O4NPs in batch experiments. Moreover, the optimum dosage of Fe3O4NPs produced hydrogen production (HP) of 4.95 L H2/d in an expanded granular sludge bed (EGSB) reactor. Fe3O4NPs involved could promote ethanol and acetic acid accumulation. Fe2+ as by-product of iron corrosion could effectively promote the activity of key coenzymes and soluble microbial products (SMPs). Importantly, Fe3O4NPs addition resulted in the formation of electronic conductor chains to enhance the electron transport efficiency in the granular sludge. Microbial community analysis revealed that the relative abundance of butyrate-hydrogen-producing bacteria (Clostridium) decreased from 40.55% to 11.45%, while the relative abundance of ethanol-hydrogen-producing bacteria (Acetanaerobacterium and Ethanoligenens) increased from 19.62% to 35.35% with Fe3O4NPs involved, confirming that the fermentation type was transformed from butyrate-type to ethanol-type, which finally facilitated more hydrogen production.

Chrysanthemum Flos is the prestigious traditional Chinese medicinal material and the popular health drink. This article comprehensively evaluated the chemical constituents, antioxidant activity, and hepatoprotective effects of 25 common chrysanthemum varieties in China. Firstly, we analyzed the chemical compositions of water extracts of chrysanthemum using UPLC/Q-TOF-MS, and identified 29 chemical components. The results displayed that chrysanthemum was rich in chemical constituents, but there were significant differences in the contents of four phenolic acids and five flavonoids among different varieties, and the coefficient of variation (CVs) ranged from 35.96 % to 114.62 %. Then, the antioxidant activities of different chrysanthemums were investigated, respectively via 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2'-azinobis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS), and Ferric Reducing Antioxidant Power (FRAP) assays. The spectrum-effect relationships between nine main components and antioxidant activities were investigated to identify the antioxidant constitutes in chrysanthemums. Meanwhile, H2 O2 -induced hepatocyte injury testing showed wide variation in cultivar antioxidant capacity, with Tongchengju (TCJ) producing the best effect (90.32 %), followed by Chuju (CJ; 85.78 %). In addition, the hepatoprotective effects of 8 mainstream varieties were determined by the model of acute alcoholic liver injury. They protected liver from injury by affecting relevant liver function and antioxidant indexes. Huangshangongju (HSG) could decrease aspartate aminotransferase (AST) activity by 39.27 % in liver tissue; Hangju-Fubaiju (HJ-FBJ), Jinsihuangju (JSH), and Chuju (CJ) significantly decreased the malondialdehyde (MDA) content of liver tissue, which reduced by more than 40 %; Jinsihuangju (JSH) of used for tea could double the content of glutathione (GSH) and had the similar effect on superoxide dismutase (SOD) as the positive group, showing significant antioxidant capacity. Therefore, this study confirmed that chrysanthemums are potential resources as antioxidants, functional foods, and medicinal materials. Importantly, it may provide a scientific support for further development and utilization of chrysanthemum, and screen excellent varieties for different demands.

Global warming is known to affect the growth, development and reproduction of insects. In this study, the larvae developmental process and endogenous hormone levels under high temperature (36 °C) stress were investigated in the lepidopteran model insect Bombyx mori (B. mori). After high temperature treatment, the duration of 5th instar larvae was shortened by 28 ± 2 h, the content of 20-hydroxyecdysone(20E) in hemolymph was significantly increased, and the transcription levels of the 20E response genes E93, Br–C, USP and E75 were up-regulated by 1.35-, 1.25-, 1.28-, and 1.27-fold, respectively. High temperature treatment also elevated the phosphorylation level of Akt and activated the downstream BmCncC/keap1 pathway, and the transcription levels of the 20E synthesis-related genes cyp302a1, cyp306a1, cyp314a1 and cyp315a1 were up-regulated by 1.12-, 1.51-, 2.17- and 1.23-fold, respectively. The transcription levels of cyp302a1 and cyp306a1 were significantly decreased in BmN cells after treatment with the double stranded RNA of BmCncC (dsBmCncC), whereas their transcription levels were significantly increased (2.15- and 1.31-fold, respectively) after treatment with the CncC agonist Curcumin. These results demonstrated that high temperature treatment promoted the metamorphosis and the BmCncC/keap1 pathway played a role in the metamorphosis of B. mori. Our results provided clues for understanding the CncC/keap1 pathway-mediated regulation of metamorphosis of Lepidopteran insects.

Chlorantraniliprole (CAP) can induce excessive calcium release from muscle of insects, causing muscle paralysis until death, and its residues in farmland can cause poisoning in Bombyx mori (B. mori), resulting in the failure of cocooning. No reports have investigated the effects of CAP exposure on detoxification enzyme activities and detoxification-related gene expression in B. mori. In the present study, we treated mulberry leaves with CAP by the leaf-dipping method, and then B. mori larvae were continuously fed with the polluted mulberry leaves. Moreover, the detoxification enzyme activities and the expressions of detoxification-related genes in the fat body of B. mori were examined. The results showed that at 24 h after CAP exposure, the activities of P450 and GST enzymes were all significantly increased, with P450 enzymes responding fastest. CarE enzyme activity was up-regulated in 24 h, and then it was decreased compared with the control group. Furthermore, the expressions of the key genes in the PI3K/Akt/CncC signaling pathway (PI3K, PDK, Akt, CncC and Keap1) at the mRNA were significantly increased. Western blotting analysis revealed that Akt was inhibited at the protein level, resulting in decreased expression of Keap1 and increased expression of CncC. These results indicated that the PI3K/Akt/CncC signaling pathway in the fat body of B. mori responded to CAP exposure and regulated the expressions of downstream detoxification enzymes, thus enhancing the detoxifying capability of B. mori.

Tembusu virus (TMUV), a highly infectious pathogenic flavivirus, causes severe egg-drop and encephalitis in domestic waterfowl, while the determinants responsible for viral pathogenicity are largely unknown. In our previous studies, virulent strain JXSP2-4 had been completely attenuated by successive passages in BHK-21 cells and the avirulent strain was designated as JXSP-310. Based on the backbone of JXSP2-4, a series of chimeric viruses were generated according to the amino acid substitutions in NS5 and their infectivities were also analyzed in cell cultures and ducklings. The results showed that the viral titers of RNA-dependent RNA polymerase (RdRp) domain-swapped cheimeric mutant (JXSP-310RdRp) in cells and ducklings were both markedly decreased compared with JXSP2-4, indicating that mutations in the RdRp domain affected viral replication. There are R543K and V711A two amino acid substitutions in the RdRp domain. Further site-directed mutagenesis showed that single-point R543K mutant (JXSP-R543K) exhibited similar replication efficacy compared with JXSP2-4 in cells, but the viral loads in JXSP-R543K-infected ducklings were significantly lower than that of JXSP2-4 and higher than JXSP-310RdRp. Surprisingly, the single-point V711A mutation we introduced rapidly reverted. In addition, qRT-PCR and Western blot confirmed that the mutations in the RdRp domain significantly affected the replication of the virus. Taken together, these results show that R543K substitution in the RdRp domain impairs the in vivo growth of TMUV, but sustaining its attenuated infectivity requires the concurrent presence of the V711A mutation.

In this study, brewery wastewater was used as the treatment in exploring the optimal conditions and maximum processing efficiency of the completely mixed anaerobic biofilm reactor (CMABR) under the conditions of hydraulic retention time (HRT) (18 h, 24 h, and 30 h) with a rotational speed (70 rpm, 100 rpm, and 130 rpm) and influent total alkalinity (TA) (20 mmol/L, 25 mmol/L, and 30 mmol/L), which was measured by the response surface methodology (RSM). The results indicated that the maximum chemical oxygen demand (COD) removal ratio was achieved under the following conditions: HRT of 21.42 h, rotational speed of 101.34 rpm, and influent TA of 25.22 mmol/L. Analysis by scanning electron microscope (SEM) showed that the microorganisms were successfully immobilized on the polyurethane fillers before the reactor began operation. High-throughput sequencing indicated that Methanothrix and Methanospirillum were the dominant contributors for COD removal in the CAMBR under these optimum conditions.

Summary 2′3′-cyclic GMP-AMP (2′3′-cGAMP), generated by cyclic GMP-AMP synthase (cGAS) under activation by cytosolic DNA, has a vital role in innate immune response via its receptor protein stimulator of interferon genes (STING) to fight viral infections and tumors. In order to have a complete understanding of biological functions of 2′3′-cGAMP, it is important to find out whether 2′3′-cGAMP has other unrevealed binding proteins present in mammalian cells and executes unknown functions. Here we report the 2′3′-cGAMP-based photoaffinity probes that capture and isolate 2′3′-cGAMP-binding proteins. These probes enable the identification of some potential 2′3′-cGAMP-binding proteins from HeLa cells. EF1A1, an essential protein regulating protein synthesis, is further validated to associate with 2′3′-cGAMP in vitro and in cells to impede protein synthesis. Thus, our studies provide a powerful approach to enable identification of the 2′3′-cGAMP interactome, discover unknown functions of 2′3′-cGAMP, and understand its physiological/pathological roles in tumor immunity and immune-related diseases.

Long non-coding RNA (lncRNA) plays an important role in many biological processes. A large number of studies have shown that predicting the associations between lncRNAs and diseases may uncover the causation of various diseases. However, experimental determination of the lncRNA-disease associations is expensive and time-consuming. Many methods are emerging, but it is still a challenge to predict the associations between lncRNAs and diseases more accurate. More and more evidences suggest that lncRNA has interaction with miRNA, which is highly related to the occurrence of cancer, gene regulation, and cell metabolism. Therefore, in this paper, we design a new method based on the interactions of lncRNAs, diseases and miRNAs. We represent the lncRNA-disease-miRNA triplet as a tensor, innovatively, and predict the potential associations between lncRNAs and diseases via tensor decomposition. First, we build lncRNAs similarity matrix by integrating semantic information and functional interactions. Second, we collect the pairwise associations between lncRNAs, diseases, and miRNAs, respectively, and integrate them into a three-dimensional association tensor. Third, we utilize the lncRNAs similarity matrix and diseases similarity matrix as auxiliary information to perform tensor decomposition with the association tensor to obtain factor matrices of lncRNAs, diseases and miRNAs, respectively. Finally, we use the factor matrices to reconstruct the association tensor for new prediction of triplet associations. To evaluate the performance of TDLDA, we compare our method with other methods and find it more superior.

Kaempferol, a flavonoid identified in a wide variety of dietary sources, has been reported to possess anti-obesity properties; however, its underlying mechanism was poorly understood. Chronic, low-grade gut inflammation and dysbacteria are proposed as underlying factors as well as novel treatment approaches for obesity-associated pathologies. This present study aims to investigate the benefits of experimental treatment with kaempferol on intestinal inflammation and gut microbial balance in animal model of obesity. High fat diet (HFD) was applied to C57BL/6J mice for 16 weeks, during which the supplement of kaempferol served as a variable. Clearly, HFD induced obesity, fat accumulation, glucose intolerance and adipose inflammation, the metabolic syndrome of which was the main finding. All these metabolic disorders can be alleviated through kaempferol supplementation. In addition, increased intestinal permeability, infiltration of immunocytes (macrophage, dendritic cells and neutrophils) and overexpression of inflammatory cytokines (tumor necrosis factor-alpha, interleukin-1beta, interleukin-6, monocyte chemoattractant protein-1) were also found in the HFD-induced mice. Kaempferol supplementation improved intestinal barrier integrity and inhibited gut inflammation, by reducing the activation of TLR4/NF-κB pathway. Furthermore, the characterization of the cecal microbiota by sequencing showed that kaempferol supplementation was able to counteract the dysbiosis associated to obesity. Our study delineated the multiple mechanism of action underlying the anti-obesity effect of kaempferol, and provide scientific evidence to support the development of kaempferol as a dietary supplement for obesity treatment.

Identifying cancer drivers is a crucial challenge to explain the underlying mechanisms of cancer development. There are many methods to identify cancer drivers based on the single mutation site or the entire gene. But they ignore a large number of functional elements with medium in size. It is hypothesized that mutations occurring in different regions of the protein sequence have different effects on the progression of cancer. Here, we develop a novel functional region driver(frDriver) identification method based on Bayesian probability and multiple linear regression models to identify protein regions that can regulate gene expression levels and have high functional impact potential. Combining gene expression data and somatic mutation data, with functional impact scores(SIFT, PROVEAN) as a priori knowledge, we identified cancer driver regions that are most accurate in predicting gene expression levels. We evaluated the performance of frDriver on the BRCA and GBM datasets from TCGA. The results showed that frDriver identified known cancer drivers and outperformed the other three state-of-the-art methods(eDriver, ActiveDriver and OncodriveCLUST). In addition, we performed KEGG pathway and GO term enrichment analysis, and the results indicated that the cancer drivers predicted by frDriver were related to processes such as cancer formation and gene regulation.

The high-temperature stress gene GrpE plays an important role in coping with high-temperature stress. The mutation of key sites of this gene can improve the high-temperature resistance of organisms. In the present study, using complementary DNAs from the silkworm fat body as the template, the open reading frame sequence of the GrpE gene (BmGrpE) was amplified and was found to be 644 bp in length and encode a protein with a predicted molecular weight of 24.1 kDa. The presence of a binding site for the heat shock transcription factor (Hsf1) at -1440 bp upstream of its coding region indicates that BmGrpE may respond to high-temperature stress. BmGrpE was constitutively expressed throughout developmental stages, with the highest level observed in the 5th instar larvae stage. Moreover, in 5th instar larvae (the 3th day), BmGrpE was expressed in all tissues examined, with the highest levels in the fat body, silk gland, and midgut. Interestingly, under high-temperature stress, TiO2 nanoparticle treatment increased the messenger RNA levels of BmGrpE in the fat body and silk gland. After treatment with dsRNA of BmGrpE, the cell viability of BmN cells was significantly decreased under 34°C and H2 O2 stress (p < .05). Mutation of BmGrpE (H163L) enhanced the resistance of BmN cells under high-temperature stress. These results provide new clues for the study of molecular mechanisms of insect resistance to high temperatures.

The massive use of fossil fuels since the industrial revolution has led to a rapid increase in the concentration of carbon dioxide (CO2) in the atmosphere. What effects elevated CO2 concentrations (ECO2) have on the defense mechanisms plants employ against insects remains poorly understood. This study showed that ECO2 of 750 ± 20 mmol/mol, increased the photosynthetic rate and biomass gain of tobacco and melon plants. However, while mass gain of Spodoptera litura, a nocturnal moth in the Noctuidae family, was higher when feeding on tobacco plants under ECO2, mass gain of Diaphania indica was reduced when feeding on melon plant at ECO2 compared to ambient CO2. Plants have many mechanisms to defend themselves against insects. Jasmonic acid (JA) is a crucial element of plant defense against lepidopteran insects. Our study showed that JA levels increased in tobacco plants under ECO2 but decreased in melon plants. It is speculated that ECO2 changes plant resistance to insects mainly by affecting the JA signaling pathway. Nutrient analysis suggested defensive metabolites rather than changes in the total nitrogen or protein content of the plants led to the changes in plant defense levels under ECO2. In summary, ECO2 affects the interaction between plants and insects. The results may provide a theoretical basis for studying the changes in crop resistance to pests under ECO2 and predicting the impact of ECO2 on future agro-ecosystems.

Salmonella infection not only causes acute and chronic diseases in poultry flocks, but the infected poultry are among the most important reservoirs for a variety of Salmonella serovars frequently transmitted to humans. This study aimed to investigate the occurrence of Salmonella spp. in local poultry farms in China. Samples (n = 4255), including dead-in-shell embryos, culled day-old-hatchings and 1- to 4-week-old diseased birds, were collected for Salmonella culture from broiler chicken, meat-type duck and pigeon farms in northern China between 2014 and 2018. A total of 103 Salmonella were isolated. S. enterica serovar Enteritidis and S. Typhimurium were the most prevalent serovars, representing 53.4% and 34.9% of the isolates, respectively. Serovar diversity was the highest in ducks, with the S. Apeyeme being isolated for the first time from duck tissues. All isolates were characterized by multilocus sequence typing (MLST) and pulsed-field gel electrophoresis (PFGE). MLST showed that all S. Enteritidis isolates shared the same sequence type (ST11), and Typhimurium showed several rare STs in addition to ST19. In comparison, PFGE showed better discrimination for S. Enteritidis and S. Typhimurium isolates, with nine distinct pulsotypes being observed. The isolates exhibited varying degrees of resistance to 15 tested antimicrobials and identified S. Enteritidis isolates (98.18%) with multiple antimicrobial resistance were a cause for concern. Our data on invasive Salmonella infection in meat-type poultry in local farms can be used to identify sources and factors associated with Salmonella spread in poultry and the associated food chain.

Silk gland is a major organ of Bombyx mori for the synthesis and secretion of silk protein. Phoxim exposure can be toxic to B. mori and causes a decrease of fibroin synthesis, finally affecting the silk production in industry. To study the mechanism of metabolism and detoxification of silk gland under phoxim exposure, we measured the residual quantity of phoxim in silk gland and hemolymph after phoxim exposure, and the detoxifying enzymes-related genes and enzyme activity were also investigated. Results indicated that the residual amount of phoxim existed up to 24 h in silk gland compared with that in hemolymph, suggesting that phoxim can accumulate in the silk glands within a certain time course. The transcriptional levels of PI3K/Akt genes, including Akt, Tor1, p70s6k and 4e-bp, were up-regulated by 6.919, 1.358, 10.766 and 7.708-fold, respectively. The expression of two downstream genes (CncC and Keap1) was up-regulated by 1.939 and 3.373-fold, respectively. In addition, the transcriptional levels of detoxification-related genes including CYP6AB, CYP306A, CarE2, GST1 and GSTd1 were up-regulated by 1.731, 1.221, 1.366, 1.376 and 6.591-fold, respectively. The enzymatic activity of CYP450, CarE and GST were increased over time. These results provided possible insights into the injury of silk gland and the transcriptional response to detoxifying enzymes-related genes in silkworm after phoxim exposure.

Ginsenosides attract great attention for their bioactivities. However, their contents are low, and many UDP-glycosyltransferases (UGTs) that play crucial roles in the ginsenoside biosynthesis pathways have not been identified, which hinders the biosynthesis of ginsenosides. In this study, we reported that one UDP-glycosyltransferase, UGTPg71A29, from Panax ginseng could glycosylate C20-OH of Rh1 and transfer a glucose moiety to Rd, producing ginsenosides Rg1 and Rb1, respectively. Ectopic expression of UGTPg71A29 in Saccharomyces cerevisiae stably generated Rg1 and Rb1 under its corresponding substrate. Overexpression of UGTPg71A29 in transgenic cells of P. ginseng could significantly enhance the accumulation of Rg1 and Rb1, with their contents of 3.2- and 3.5-fold higher than those in the control, respectively. Homology modeling, molecular dynamics, and mutational analysis revealed the key catalytic site, Gln283, which provided insights into the catalytic mechanism of UGTPg71A29. These results not only provide an efficient enzymatic tool for the synthesis of glycosides but also help achieve large-scale industrial production of glycosides.

Folate metabolism pathway is mainly associated with syntheses of nucleic acids and proteins as well as DNA methylation and repair. Cytoplasmic C1 tetrahydrofolate (C1-THF) synthase is a central enzyme in folate metabolism and plays an important role in C1-THF conversion. The full-length sequence of silkworm (Bombyx mori) cytoplasmic C1-THF synthase (BmC1-THF synthase) gene including an open reading frame (ORF) of 932 amino acid residues was cloned. Phylogenetic analysis indicated that BmC1-THF synthase shares 84% and 81% identity with C1-THF synthase of Papilio Xuthus and Danaus plexippus, respectively. Conserved sequence analysis showed that the 330 to 710 amino acid residues of BmC1-THF synthase were highly conserved among various organisms. Recombinant BmC1-THF synthase protein with a molecular weight of about 105.5 kDa was expressed in E. coli cells and identified using SDS-PAGE and western blot analysis. The expression of BmC1-THF synthase gene in eight silkworm tissues revealed that BmC1-THF synthase gene is widely expressed in various tissues of silkworm and highly expressed in the fat body. The expression characteristics of BmC1-THF synthase gene in response to oxidative stress were conducted under phoxim and high temperature (30 °C) exposure. The results showed that the transcriptional level of BmC1-THF synthase gene was upregulated by 3.435-fold and 6.845-fold after phoxim and high temperature (30 °C) exposure, respectively, indicating that BmC1-THF synthase gene may play a significant role in response of silkworm to oxidative stresses.

Silkworm, Bombyx mori (L.; Lepidoptera: Bombycidae), is an economically important insect, which is sensitive to the environment and susceptible to oxidative damages at high temperature. Low concentrations of TiO2 NPs (titanium dioxide nanoparticles) can scavenge reactive oxygen species (ROS) produced by oxidative damages in vivo. To explore whether TiO2 NPs could alleviate oxidative damages of high temperature, the effects of TiO2 NPs treatment on silkworm growth, the levels of ROS and H2O2, as well as the transcription level of antioxidant-related genes were studied at 30°C. These results showed that TiO2 NPs treatment increased silkworm body weight by 6.0% and reduced the occurrence of irregular cocoon at 30°C. TiO2 NPs treatment at 30°C decreased ROS levels in fat body and increased expression of Hsp70, SOD by 5.70-fold at 48 h, TPx by 1.61-fold, CAT by 1.81-fold. These results indicated that TiO2 NPs treatment at 30°C could promote the expression of antioxidant genes and reduce oxidative stress and provide a new method to alleviate high-temperature induced oxidative stress to silkworm.

This study reports the best culture conditions for roots growth and accumulation of active components by optimizing the parameters. Glycyrrhiza uralensis adventitious roots metabolites were significantly increased after adding Saccharomyces cerevisiae and Meyerozyma guilliermondii. The highest contents of polysaccharide, glycyrrhizic acid, glycyrrhetinic acid, and total flavonoids were obtained in M. guilliermondii group; the content of glycyrrhizic acid was 5.3-fold higher than the control. In control and treatment groups, 12 compounds were identified by high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry (HPLC-ESI-MS/MS), among which some new compounds have been detected in elicitor groups including 5,7-dihydroxyflavanone, glycyrrhisoflavanone, licorice saponin J2, uralsaponin B, (3R)-vestitol, and uralenol. Meyerozyma guilliermondii significantly upregulated the expression of the genes such as 3-hydroxy-3-methylglutaryl coenzyme A reductase, farnesyl diphosphate synthase, geranyl diphosphate synthase, squalene synthase, squalene epoxidase, β-amyrin synthase, and CYP88D6 and CYP72A154. Meanwhile, it increased the biosynthesis of signaling molecules (nitric oxide, salicylic acid, and jasmonic acid) in defense mechanism.

microRNA 21 (miRNA‑21) promotes the development of cardiac fibrosis, hypertrophy and heart failure. However, whether it can be used as a biomarker for the diagnosis and prognosis of heart failure remains unclear. The current study assessed circulating miRNA‑21 as a viable indicator for diagnosis and prognosis of heart failure. The levels of miRNA‑21 and brain natriuretic peptide were measured in serum obtained from the peripheral vein (miRNA‑21‑PV) and coronary sinus (miRNA‑21‑CS) of 80 patients with heart failure and 40 control individuals via reverse transcription‑quantitative polymerase chain reaction and ELISA, respectively. The correlations between circulating miRNA‑21 and diagnosis, severity, prognosis and re‑hospitalization rate of heart failure were evaluated using statistical analysis. Serum miRNA‑21‑PV and miRNA‑21‑CS levels of patients with heart failure were significantly higher than that of control subjects, and were also correlated with ejection fraction and brain natriuretic peptide. Both were determined to have high levels of sensitivity and specificity for diagnosing heart failure. Follow‑up of the patients with heart failure indicated that miRNA‑21‑PV and miRNA‑21‑CS were correlated with prognosis, and miRNA‑21‑CS was efficient in predicting re‑hospitalization for heart failure. Circulating miRNA‑21 has potential to be a biomarker of heart failure.

The protein tyrosine phosphatase 1B (PTP1B) is an important regulator of metabolism. The relationship between PTP1B and tumors is quite complex. The purpose of this study is to explore the expression pattern and role of PTP1B in breast cancer. The expression of PTP1B was detected in 67 samples of breast cancer tissue by Western blot. Cell growth assay, Transwell migration assay, and Scratch motility assay were used to examine the proliferation and migration of MCF-7 with and without PTP1B. The total levels and phosphorylated levels of signal transduction and activator of transcription 3 (STAT3) and the expression of C-C motif chemokine ligand 5 (CCL5) were also examined by Western blot. PTP1B was overexpressed in over 70% of breast cancer tissues, correlating with patients with estrogen receptor (ER)-negative, progesterone receptor (PR)-negative, and human epidermal growth factor receptor 2 (HER2)-positive tumors. The data also showed that both tumor size and lymph node metastasis were significantly higher in patients with a higher level of PTP1B. The proliferation and migration of MCF-7 cells were found to be inhibited after knocking down the gene of PTP1B. Our data also showed that PTP1B could up-regulate the dephosphorylated level of STAT3, which could increase the expression of CCL5. These phenomena indicated that PTP1B may play a crucial role in the development of breast cancer.

Over past decades plant tissue culture has emerged as an alternative of whole plant cultivation in the production of valuable secondary metabolites. Adventitious roots culture of Panax ginseng and Echinacea purpure has reached the scale of 1-10 kL. Some molecular biological techniques, such as transgenic technology and genetic stability are increasingly used in the studies on plant tissue cultures. The studies on elicitors have deepened into the induction mechanism, including signal molecules, functional genes, and so on. More and more biological elicitors, such as A. niger and yeast are used to increase the active compounds in plant tissue cultures. We also discussed the application of synthetic biology in the studies on biosynthesis of artemisinin, paclitaxel, and tanshinon. The studies on active ingredients biosynthesis of medicinal plants provide unprecedented possibilities to achieve mass production of active ingredients. Plant tissue cultures can not only produce active ingredients but also as experimental materials for biosynthesis. In order to improve the contents of active compounds in medicinal plants, following aspects could be carried out gene interference or gene silencing, gene overexpression, combination with chemical synthesis, application of elicitors, and site-directed mutagenesis of the key enzymes.

This research explored the effects of protein and polysaccharide in Meyerozyma guilliermondii on active compounds in Glycyrrhiza uralensis Fisch adventitious roots. In this study, a responsive protein LSP1 was purified from the Meyerozyma guilliermondii since the excellent induction. The contents of total flavonoids (3.46 mg · g−1), glycyrrhizic acid (0.41 mg · g−1), glycyrrhetinic acid (0.41 mg · g−1) and polysaccharide (94.49 mg · g−1) in adventitious root peaked at LSP1 group, which were 1.6, 3.4, 2.4, 2.0-fold that of control, respectively. Besides, the responsive protein LSP1 significantly activated the defense signaling, mitogen-activated protein kinases and extremely up-regulated the expression of defense-related genes and functional genes involved in glycyrrhizic acid biosynthesis. This article is protected by copyright. All rights reserved

In this work, we selected three fungi strains (Aspergillus niger, Aspergillus flavus and Aspergillus oryzae) as elicitors prepared from mycelium or fermentation broth to improve ginsenosides production in adventitious roots culture. The results showed that ginsenosides production (29.90 ± 4.67 mg g−1) was significantly enhanced upon elicitation with 200 mg L−1 A. niger elicitor prepared from mycelium, which was 3.52-fold of untreated group. HPLC-ESI–MSn analysis was performed, showing that ginsenoside Rb3 was present after treatment with the A. niger. Furthermore, we found that A. niger significantly enhanced accumulation of Nitric oxide (NO), salicylic acid (SA) and jasmonic acid (JA) involved in plant defense response, and significantly up-regulated the expression of the geranyl diphosphate synthase (GPS), farnesyl diphosphate synthase (FPS), squalene synthase (SS), squalene epoxidase (SE), dammarenediol synthase (DS), Two cytochrome P450 (CYP) genes (CYP716A47 and CYP716A53v2) and three UDP-glycosyltransferases (UGT) genes (UGTAE2, UGT94Q2 and UGTpg100).

Magnetite nanoparticles (Fe3O4 NPs) were applied in an anaerobic semi-continuous tank reactor (ASTR) to investigate its effect on the anaerobic digestion (AD) of acetate synthetic wastewater. The Fe3O4 NPs corrosion could create a more favorable micro-environment to enhance the methanogens activity. The chemical oxygen demand (COD) removal efficiency and methane production in test (ASTRT) were 31.1% and 101.5% higher than those in control (ASTRC). With the addition of Fe3O4 NPs, the concentration of key coenzyme (F420 and M) increased from averagely 0.523 and 5.352 μmol/g-VSS to 0.956 and 9.267 μmol/g-VSS, and the content of soluble microbial products (SMPs) significantly increased. Additionally, the high-throughput 16S rRNA gene sequencing further confirmed that the percentage of hydrogen-utilizing methanogens (Methanolinea) was up to 62.6% of total archaeal sequences. Fe3O4 NPs addition would accelerate electrons transfer from acids oxidizers to syntrophic methanogenesis, further stimulate acids oxidizers to decompose acetate to H2/CO2, and finally facilitate more methane production.

Silkworm (Bombyx mori) is an economic insect of the Lepidoptera. Chlorantraniliprole (CAP) exposure results in reduced growth and development of B. mori and failure in cocooning, seriously affecting the development of sericulture. To study the mechanisms underlying the damage to silkworm caused by sublethal doses of CAP, we examined the oxidative damage, the activities of digestive enzymes in midgut, and the expressions of midgut-related genes at the mRNA level. We found that CAP exposure inhibited the growth of silkworm, decreased the body mass and caused the accumulation of reactive oxygen species (ROS) [the levels of O2-, H2O2 and lipid peroxidation (MDA) were increased by 1.62-, 1.87- and 1.46-fold, respectively]. Moreover, we also found that the midgut cells were disintegrated, microvilli disappeared, the stroma became thinner, and the chromatin of nucleus became aggregated after CAP exposure by the analysis of transmission electron microscopy (TEM). In addition, the activities of digestive enzymes were dysregulated in midgut (the activities of α-amylase and trypsin were decreased 0.69- and 0.20-fold, respectively). Furthermore, digital gene expression (DGE) profiling analysis revealed that the expressions of oxidative phosphorylation pathway and antioxidant defense system related genes in midgut were decreased, indicating that it was the oxidative damage in midgut caused by CAP that mainly affected the growth of silkworm, rather than the toxicological effects of CAP. Collectively, this study provided valuable insights into the toxic effects of CAP on insects.

Carbon starvation is the current leading hypothesis of plant mortality mechanisms under drought stress; recently, it is also used to explain tree die-off in plant diseases. However, the molecular biology of the carbon starvation pathway is unclear. Here, using a punch inoculation system, we conducted transcriptome and physiological assays to investigate pathogen response in poplar stems at the early stages of Botryosphaeria and Valsa canker diseases. Transcriptome assays showed that the majority of differentially expressed genes (DEGs) in stem phloem and xylem, such as genes involved in carbon metabolism and transportation, aquaporin genes (in xylem) and genes related to the biosynthesis of secondary metabolites and the phenylpropanoid pathway (related to lignin synthesis), were downregulated at 7 days after inoculation (DAI). Results also showed that the expression of the majority of disease-resistance genes upregulated in poplar stems, which may be connected with the downregulation expression of the majority of WRKY family genes. Physiological assays showed that transpiration rate decreased but WUE (water use efficiency) increased the 3 and 7 DAI, while the net photosynthetic rate decreased at 11 DAI in Botryosphaeria infected poplars (ANOVA, P

Summary Aims There is currently no specific treatment for non-alcoholic fatty liver disease (NAFLD) in children, but recent studies have shown that vitamin E may be effective. Therefore, we conducted a meta-analysis of trials in which vitamin E was used to treat paediatric NAFLD. Methods We searched the PubMed, Embase, Scopus and Cochrane Library databases to identify related articles published prior to March 2020 that examined the effect of vitamin E for the treatment of paediatric NAFLD. Results The results showed that vitamin E significantly decreased low-density lipoprotein (LDL) and total cholesterol (TCHO) levels. However, no significant changes were found in other indicators, including body mass index (BMI), triglyceride (TG) levels, high-density lipoprotein (HLD) levels, fasting insulin levels, homeostatic model assessment (HOMA-IR), alanine transaminase (ALT) levels, aspartate aminotransferase (AST) levels, glutamate transpeptidase (GGT) levels, ballooning degeneration and fibrosis (P > 0.05). Although the P value of NAS was less than 0.05, the evidence was not strong enough. We also found that treatment with vitamin E significantly increased fasting glucose (FSG) levels if the intervention time was ≤12 months. Conclusions Vitamin E therapy can improve blood lipids to some extent, but its effect on children's liver function and liver tissue is not apparent, and the finding that this therapy increases FSG levels still needs more research. Systematic review registration PROSPERO CRD42020177663

White Chrysanthemum (Chrysanthemum morifolium), a perennial herb of the Compositae family, is used for traditional medicine. The planting area of white chrysanthemum in Macheng city, Hubei Province is about 3333 ha and the annual output can reach more than 5000 tons. In 2019, leaf spot disease appeared on almost all middle and lower leaves of white chrysanthemum in most fields of Fengshumiao county, Macheng city (N31°29'57″, E115°05'49″). This county has 33 acres white chrysanthemum planting area, and most of the plants in the county were infected with the leaf spot disease. The average incidence of leaf spot disease was 65%, and incidence in some areas was 100%. In our observations, leaf spot disease can occur throughout the whole growth period of white chrysanthemum, and it will become more serious under the high temperature and humidity condition. Usually, the diseased leaves account for 30 to 80% of the total leaves on the plant. Leaf spot initially manifests as necrotic lesions on the edge and tip of the leaf, and then the lesions coalesce and gradually expand to form irregular light-brown to brown-black spots, eventually leading to necrosis and curling of the entire leaf. This disease seriously affects the growth and development of plants, resulting in the decline of yield and quality of white chrysanthemum. Ten symptomatic leaf samples were collected, the surfaces were disinfected with 0.1% mercuric chloride (HgCl2) for 3 min, and washed with sterile distilled water three times. Ten tissue samples at the junction of diseased and healthy areas (0.5 × 0.5 cm2) were cut and placed on potato dextrose agar (PDA) medium containing 100 µg/ml cefotaxime sodium and incubated in a dark chamber at 28°C. After 2 days, the hyphal tips from the edges of growing colonies were transferred to fresh PDA plates for further purification. Finally, eight isolates were obtained and these isolates were similar in morphology. The color of purified isolates was initially white to pale yellow. After six days of incubation, colonies had a diameter of 8 cm and the cultures were pale gray and starting to secrete scarlet pigment. After 15 days incubation, the colonies were grayish brown, while the backside was reddish-brown. Gray to tan chlamydospores were observed, nearly spherical, with a wart-like surface. Unicellular chlamydospores were 7.91 to 32.23 × 12.03 to 38.42 µm (n=30) and multicellular chlamydospores were 6.32 to 25.10 × 21.75 to 100.05 µm (n=30). The morphological characteristics were similar to Epicoccum sorghinum (Kang et al. 2019). The isolate FDY-5 was chosen for molecular identification. The sequence of rDNA-ITS, TUB, and LSU of the FDY-5 were amplified (GenBank MT800929, MT799852, and MT800935, respectively) (White et al. 1990; Carbone and Kohn 1999; Lumbsch et al. 2000). BLAST results showed that the rDNA-ITS sequences, the TUB gene sequences, and LSU gene sequences of strain FDY-5 shared 99% identity with the sequences of E. sorghinum (syn. Phoma sorghina) in GenBank (MN555348.1, MF987525.1, MK516207.1, respectively). Moreover, a phylogenetic tree of the LSU gene sequence of FDY-5 was constructed based on the Neighbor-Joining (NJ) method in MEGA6 software (Tamura et al. 2013) and revealed that strain FDY-5 was closest to E. sorghinum. Based on morphological and molecular characteristics, the fungus was identified as E. sorghinum. Pathogenicity tests were conducted on two-month-old white chrysanthemum plants. The upper three leaves of three plants were randomly selected for stab treatment and were inoculated with 5 × 5 mm mycelial discs produced from a fifteen-day-old colony on PDA. The inoculated and control (treated with sterile PDA disks) plants were incubated in a moist chamber (25 ± 2 °C, RH 85%). The first lesions appeared 1 day after inoculation on leaves, and the necrotic lesion area expanded outward and showed typical symptoms 3 days later. To fulfill Koch's postulates, the pathogen was reisolated from nine inoculated leaves by repeating the above isolating operation, and confirmed as E. sorghinum by morphology. To the best of our knowledge, this is the first report of E. sorghinum causing leaf spot on white chrysanthemum in China. E. sorghinum has a wide host range worldwide and often causes crop yield reduction. This report will facilitate the diagnosis of white chrysanthemum leaf spot of white chrysanthemum allowing control measures to be adopted to manage this disease in a timely manner. References Carbone, I., and Kohn, L. M. 1999. Mycologia 91:553. Kang, Y., et al. 2019. Plant Dis. 103 (7):1787. Lumbsch, H., et al. 2000. Plant Biol. 2:525. Tamura, K., et al. 2013. Mol. Biol. Evol. 30:2725-2729. White, T. J., et al. 1990. Page 315 in:PCR protocols:a guide to methods and applications. Academic Press, San Diego, CA. Funding Funding was supported by Major Increase and Decrease Projects at the Central Level of China (2060302) and the National Key Research and Development Program (2017FYC1700704).

Magnetite nanoparticles (Fe3O4 NPs) was firstly used to enhance pollutants removal during coal gasification wastewater (CGW) treatment in anaerobic digestion (AD) system. Bench-scale results revealed that 200 mg/L and 20–40 nm of Fe3O4 NPs addition resulted in a maximum removal capacity of total phenol (TPh) at a temperature of 36 °C and hydraulic retention time (HRT) of 36 h. Meanwhile, Fe3O4 NPs addition reduced the oxidation reduction potential (ORP) values and biological toxicity, and enhanced the stability of AD system. Pilot-scale results showed that the TPh and chemical oxygen demand (COD) removal efficiency (53% and 49%) were obtained with the optimal dosage of Fe3O4 NPs. Moreover, electron nanowires may be established with Fe3O4 NPs assisted to perform direct interspecies electron transfer (DIET) among Geobacter, Pseudomonas and Methanosaeta species, and finally enhanced the pollutants removal efficiency.

Tree canker diseases are mainly caused by necrotrophic fungal pathogens, which induce canker lesions, wilting, or dieback in many species. We hypothesized that canker-associated canopy dieback is related to functional failure of the phloem and cambium, carbon starvation and/or hydraulic failure. Using a girdling and inoculation system, we evaluated gas exchange and hydraulic parameters, concentrations of non-structural carbohydrates (NSCs), leaf characteristics, stem phenotyping and symptoms of 6-month-old poplar saplings with Botryosphaeria or Valsa canker disease. Pathogen inoculations inhibited callus formation and phloem regeneration and altered the physiological parameters, water potential, NSC content in stems, and symptoms on leaves and stems, while the girdling-only control did not. Results suggest that the functional and structural integrity of the phloem and cambium play crucial roles in the pathogenesis of canker disease. Botryosphaeria canker decreased photosynthetic rate (−54% to −75%), stomatal conductance (−71% to −80%) and transpiration rate (−58% to −66%), while Valsa canker decreased photosynthetic rate (−53% to −83%), stomatal conductance (−71% to −87%) and transpiration rate (−53% to −76%) at 20, 25, and 30 days after inoculation. Results suggest that the two fungal pathogens induce carbon starvation by inhibiting photosynthesis. The leaf water potential was higher and the change in specific hydraulic conductivity was smaller in the canker region than in the adjacent regions, while the NSC content was lower in the region below cankers than above them, suggesting that the fungal pathogens caused poplar canopy dieback by initially inducing carbon starvation, not hydraulic failure.

In this study, Candida tropicalis, Aspergillus niger, and a mixture of their cultures were previously used as fungal elicitors in Panax quinquefolium adventitious root culture to enhance ginsenoside production. The result showed that ginsenoside content (11.47 ± 0.54 mg g−1) was significantly enhanced to 3.25-fold of the control group after treatment with 400 mg L−1 C. tropicalis. The polysaccharide content (213.01 ± 18.63 mg g−1) reached the highest when A. niger was at the concentration of 200 mg L−1. HPLC–ESI-MSn analysis was performed, showing that ginsenosides Rh1 and Ro were identified after treatment with fungal elicitors. Furthermore, we found that C. tropicalis, A. niger, and mix elicitor significantly up-regulated the expression of the squalene synthase, squalene epoxidase, dammarenediol synthase, β-amyrin synthase, protopanaxadiol synthase, protopanaxatriol synthase, and UDP-glycosyltransferase. SDS-polyacrylamide gel electrophoresis was used to analysis the changes of protein. A new band was discovered at the size of 75 kDa. In addition, it found that fungal elicitors significantly increased the activities of catalases and peroxidase.

Spinocerebellar ataxia type 3 (SCA3) is a rare inherited autosomal dominant progressive neurological disorder, which results from a CAG‑repeat expansion in the gene encoding the deubiquitinating enzyme, ataxin‑3. At present, no effective treatment is available for this fatal disorder; however, certain studies have suggested that reducing the levels of mutant ataxin‑3 protein may reverse or halt the progression of disease in patients with SCA3. In the present study, clinical examinations were performed on a patient with SCA3 who exhibited disease features including coughing, expectoration and was bedridden with mobility limitation. CAG repetitions at SCA‑associated genes were detected in the patient's family by performing standard polymerase chain reaction (PCR) and triple‑repeat primed PCR. The numbers of CAG‑repeats within the two alleles of the gene of interest in the patient were 15 and 78. Notably, the patient's brother, who harbored 76 CAG‑repeats in one allele of the gene of interest, did not exhibit severe disease symptoms. These results suggest that the number of CAG‑repeats is a critical for determination of SCA3 disease severity and time of onset. In addition, the defined phenotypic characteristics of the patient in the present study provide useful insight for more accurate clinical diagnosis and genotyping of future patients.

Panax notoginseng, an important herbal medicine, has wide uses for its bioactive compounds and health function. In this work, we compared the content of saponin in cultivation and adventitious root. The total content of saponins in adventitious root (8.48 mg⋅g-1 ) was found lower than in the native one (3-year-old) (34.34 mg⋅g-1 ). To enhance the content of bioactive compounds, we applied elicitors jasmonic acid (JA) and methyl dihydrojasmonate (MDJ) to the adventitious root culture. It was observed that the highest total content of saponins (71.94 mg⋅g-1 ) was achieved after treatment with 5 mg⋅L-1 JA, which was 2.09-fold higher than native roots and 8.45-fold higher than the control group. The findings from high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry analysis showed that six new compounds were present after the treatment with the elicitors. Furthermore, we found that JA and MDJ significantly upregulated the expression of the geranyl diphosphate synthase, farnesyl diphosphate synthase, squalene synthase, squalene epoxidase, dammarenediol synthase, and CYP716A47 and CYP716A53v2 (CYP450 enzyme) genes; downregulated the expression of the cycloartenol synthase gene; and increased superoxide dismutase and peroxidase activities.

This study explored the ability of three rhizobacterial strains (Bacillus subtilis, Penicillium fellutanum and Escherichia coli) to trigger metabolism. The protein fragment of more than 10 kDa significantly increased the metabolite contents in Glycyrrhiza uralensis adventitious roots. The results showed the highest accumulation of total flavonoids (7.59 mg g−1), glycyrrhizic acid (0.29 mg g−1), glycyrrhetinic acid (0.27 mg g−1) and polysaccharide (93.11 mg g−1) by up to 2.27-fold, 2.64-fold, 2.70-fold and 2.32-fold that of control roots, respectively. Besides, the protein fragment of more than 10 kDa significantly activated defense signaling and extremely up-regulated the expression of defense-related genes and functional genes in glycyrrhizic acid and flavonoid biosynthesis. In Glycyrrhiza uralensis adventitious roots, HPLC-ESI-MSn analysis showed that the protein fragment of more than 10 kDa induced the generation of four new compounds over the control group.

The global warming has affected the growth, development and reproduction of insects. However, the molecular mechanism of high temperature stress-mediated metamorphosis regulation of lepidopteran insect has not been elucidated. In this study, the relationship between the insect developmental process and endogenous hormone level was investigated under high temperature (36 ° C) stress inBombyx mori(B. mori). The results showed that the duration of 5thinstar larvae were shortened by 28 ± 2 h, and the content of 20E was up-regulated significantly after 72 h of high temperature treatment, while the transcription levels of 20E response genesE93, Br-C, USP, E75were up-regulated 1.35, 1.25, 1.28, and 1.27-fold, respectively. The high temperature treatment promoted the phosphorylation level of Akt and the downstream BmCncC/keap1 pathway was activated, the transcription levels of 20E synthesis-related genescyp302a1, cyp306a1, cyp314a1andcyp315a1were up-regulated by 1.12, 1.51, 2.17 and 1.23-fold, respectively. After treatment with double stranded RNA of BmCncC (dsBmCncC) in BmN cells, the transcription levels ofcyp302a1andcyp306a1were significantly decreased, whereas up-regulated by 2.15 and 1.31-fold, respectively, after treatment with CncC activator Curcumin. These results suggested that BmCncC/keap1-mediated P450 genes (cyp302a1, cyp306a1) expression resulted in the changes of endogenous hormone level, which played an important role in the regulation of metamorphosis under high temperature stress. Studies provide novel clues for understanding the CncC/keap1 pathway-mediated metamorphosis regulation mechanism in insects.Author SummaryMammalian nuclear transcription factor Nrf2 (NF-E2-related factor 2) plays an important role in the stress response of cells. CncC is a homolog of mammalian Nrf2 in insect, regulating the genes expression of insect antioxidant enzymes and cytochrome P450 detoxification enzyme. Evidence suggests that the CncC/Keap1 pathway also plays an important role in regulating insect development. Here, we investigated the regulatory mechanism between the CncC/Keap1 pathway and metabolism of silkworm hormones in Lepidoptera. We found that high temperature induction accelerated the development of silkworm, the ecdysone content and related metabolic genes in hemolymph were significantly up-regulated, the CncC/Keap1 pathway was activated, and the expression ofBmCncCwas significantly increased, indicating that the Cncc/Keap1 pathway plays an important role in this process. The expression ofcyp302a1andcyp306a1was significantly decreased by RNA interference withBmCncC, which indicated that CncC in silkworm had a regulatory relationship with downstream 20E synthetic gene. In summary, the results indicate that the CncC/Keap1 pathway plays an important role in regulating hormone metabolism in silkworm, providing a basis for further study of the relationship between CncC/Keap1 pathway and development in insects.

Various nanoparticles, such as silver nanoparticles (AgNPs) and titanium nanoparticles (TiO2 NPs) are increasingly used in industrial processes. Because they are released into the environment, research into their influence on the biosphere is necessary. Among its other effects, dietary TiO2 NPs promotes silk protein synthesis in silkworms, which prompted our hypothesis that TiO2 NPs influence protein kinase B (Akt)/Target of rapamycin (Tor) signaling pathway (Akt/Tor) signaling in their silk glands. The Akt/Tor signaling pathway is a principle connector integrating cellular reactions to growth factors, metabolites, nutrients, protein synthesis, and stress. We tested our hypothesis by determining the influence of dietary TiO2 NPs (for 72 h) and, separately, of two Akt/Tor pathway inhibitors (LY294002 and rapamycin) on expression of Akt/Tor signaling pathway genes and proteins in the silk glands. TiO2 NPs treatments led to increased accumulation of mRNAs for Akt, Tor1 and Tor2 by 1.6-, 12.1-, and 4.8-fold. Dietary inhibitors led to 2.6- to 4-fold increases in mRNAs encoding Akt and substantial decreases in mRNAs encoding Tor1 and Tor2. Western blot analysis showed that dietary TiO2 NPs increased the phosphorylation of Akt and its downstream proteins. LY294002 treatments led to inhibition of Akt phosphorylation and its downstream proteins and rapamycin treatments similarly inhibited the phosphorylation of Tor-linked downstream proteins. These findings support our hypothesis that TiO2 NPs influence Akt/Tor signaling in silk glands. The significance of this work is identification of specific sites of TiO2 NPs actions.

Bombyx mori is an economic insect of the Lepidoptera. Its posterior silk gland (PSG) is an important organ for fibroin synthesis. In order to study the occurrence of apoptosis in PSG and the role of PI3K/Akt signaling pathway during spinning period, changes in morphology of silk gland, expressions of fibroin components Fib-H, Fib-L and P25 and Akt, TOR2, P70S6K and S6 in PI3K/Akt pathway, expressions of apoptosis related genes caspase-3, caspase-9 and activity of caspase-3 were explored. The results showed that the morphology of silk gland dramatically degenerated; transcription of Fib-H, Fib-L, and P25 gradually declined with time; and Fib-L protein level reduced by 0.6-fold at 72 h. Moreover, the transcription levels of Akt, TOR2, P70S6K, and S6 also decreased by 0.3-, 0.8-, 0.7-, and 0.1-fold, respectively, indicating that the downregulation of PI3K/Akt signaling pathway could lead to reduction in fibroin synthesis. In addition, the transcription levels of caspase-3 and caspase-9 increased by 1.3- and 3.6-fold, respectively, and the enzyme activity of caspase-3 grew at a maximum of 1.6-fold. The results showed the occurrence of apoptosis in PSG during spinning period. In conclusion, the present study indicated that both the decline in fibroin components and the increase in apoptosis-related genes were regulated by PI3K/Akt signaling pathway during spinning period, which shed new light on the functions of PI3K/Akt signaling pathway.

Paramyotonia congenita (PC) is a rare autosomal dominant neuromuscular disorder characterized by juvenile onset and development of cold-induced myotonia after repeated activities. The disease is mostly caused by genetic mutations of the sodium channel, voltage-gated, type IV, alpha subunit (SCN4A) gene. This study intended to systematically identify the causative genetic variations of a Chinese Han PC family. Seven members of this PC family, including four patients and three healthy controls, were selected for whole exome sequencing (WES) using the Illumina HiSeq platform. Sequence variations were identified using the SoftGenetics program. The mutation R1448C of SCN4A was found to be the only causative mutation. This study applied WES technology to sequence multiple members of a large PC family and was the first to systematically confirm that the genetic change in SCN4A is the only causative variation in this PC family and the SCN4A mutation is sufficient to lead to PC.

In this study, adventitious roots of Glycyrrhiza uralensis were cultured in a 5-L balloon-type bubble bioreactor with different medium salt strength and different concentrations of sucrose. The results of the culture showed that the best culture conditions were 4 % sucrose concentration and 1 MS medium for the accumulation of secondary metabolites. Salicylic acid has previously been used as an elicitor in tissue cultures to enhance production of secondary metabolites. Addition of 1 mg L−1 salicylic acid significantly enhanced the contents of glycyrrhizic acid (0.31 mg g−1), glycyrrhetinic acid (0.14 mg g−1) and polysaccharide (159.29 mg g−1) in the G. uralensis adventitious roots and the contents were 2.58-fold, 1.27-fold, and 2.07-fold over the control group. Furthermore, the greatest content of total flavonoid (9.40 mg g−1) was observed with 2 mg L−1 salicylic acid added in the culture, which was 2.68-fold higher than the control. The ESI-MSn analysis was also performed, showing that two new kinds of flavonoids, including (3R)-vestitol and glycyrol, were only identified in the salicylic acid treatment. In addition, we also found that salicylic acid significantly increased superoxide dismutases, catalases, and peroxidase activity.

Mutations in the BRCA1/2 genes are associated with an increased risk of breast cancer, but no large-scale research have examined the BRCA1/2 mutations in Chinese Kazakh women. We evaluated the frequency and distributions of BRCA1 and BRCA2 gene mutations in Kazakh sporadic breast cancer patients and healthy women in China. The association between the clinical-pathologic features of Kazakh breast cancer patients and BRCA1/2 mutations were also investigated. Two unclassified variants (T539M and T1915M) and 16 polymorphisms were detected in this study, 4 of which (G356A, His743, Asn991Asp, Val1269) were detected more frequently in breast cancer patients than in healthy controls. We observed a higher prevalence of BRCA1/2 common sequence alterations and a large number of Kazakh women carrying multiple co-existing BRCA1/2 mutations. The prevalence of BRCA1 mutations was similar to that of BRCA2 mutations. Although no significant differences were observed, BRCA1/2 carriers were generally younger at diagnosis of wild-type breast cancer patients. BRCA1-associated Kazakh sporadic breast cancers present with high tumor grade, early stage, negative lymph node status, absence of estrogen receptor expression and progesterone-positive status. Estrogen receptor expression was the only predominant histological type in BRCA2 carriers. In this study, we determined the BRCA1 and BRCA2 gene mutation status and determined the association with clinical-pathologic characteristics in a Chinese Kazakh population. Larger population-based screening studies screening the entire coding region of BRCA1/2 are required to evaluate the breast cancer risk induced by the sequence alterations detected in this study.

Aminoacyl-tRNA synthetases are the key enzymes for protein synthesis. Glycine, alanine, serine and tyrosine are the major amino acids composing fibroin of silkworm. Among them, the genes of alanyl-tRNA synthetase (AlaRS) and glycyl-tRNA synthetase (GlyRS) have been cloned. In this study, the seryl-tRNA synthetase (SerRS) and tyrosyl-tRNA synthetase (TyrRS) genes from silkworm were cloned. Their full length are 1709 bp and 1868 bp and contain open reading frame (ORF) of 1485 bp and 1575 bp, respectively. RT-PCR examination showed that the transcription levels of SerRS, TyrRS, AlaRS and GlyRS are significantly higher in silk gland than in other tissues. In addition, their transcription levels are much higher in middle and posterior silk gland than in anterior silk gland. Moreover, treatment of silkworms with phoxim, an inhibitor of silk protein synthesis, but not TiO2 NP, an enhancer of silk protein synthesis, significantly reduced the transcription levels of aaRS and content of free amino acids in posterior silk gland, therefore affecting silk protein synthesis, which may be the mechanism of phoxim-silking disorders. Furthermore, low concentration of TiO2 NPs showed no effect on the transcription of aaRS and content of free amino acids, suggesting that TiO2 NPs promotes silk protein synthesis possibly by increasing the activity of fibroin synthase in silkworm.

Mounting evidence suggests that all organisms at the cellular level respond to stress by synthesizing heat shock proteins at the expense of other proteins, and the ability of human cells to respond to heat stress decreases with aging. We thus investigate the association of 3 variants (A1267G in HSPA1B , G190C in HSPA1A , and T2437C in HSPA1L ) in the heat shock protein 70 (Hsp70) family with natural longevity in a Xinjiang Hetian Uygur population. A case-control study was conducted in 191 healthy individuals greater than 90 years of age, and 53 naturally died persons 65–70 years of age. Promoter activity was evaluated by luciferase reporter assays. The data were analyzed using an EH/EH+ program for haplotype prediction and MDR software for gene–gene interaction. All studied variants satisfied the Hardy-Weinberg equilibrium in each group. In single-locus analysis, no significant differences were found between long-lived people and short-lived people in the genotype/allele distributions of all variants. In contrast, haplotype analysis indicated that haplotypes A-G-C and A-C-T were more prevalent in long-lived people than short-lived people ( P = 0.026 and 0.017), and the analysis conferred a 3.46- and 4.51-fold increased tendency for longevity, respectively ( P = 0.025 and 0.016). The haplotype results were strengthened by interaction analysis, which suggests an optimal model in which G190C and T2437C exert an interacting effect on longevity. No functional significance was observed between 190G and 190C alleles in both control and heat-inducible A549 cells ( P > 0.05). Taken together, our findings suggested that common genetic variants in Hsp70 family might contribute interactively to longevity the Xinjiang Hetian Uygur population.

Experimental evidence suggesting that heat shock protein 70 (Hsp70) gene or associated genes are responsible for the pathophysiology of hypertension is accumulating. In this study, we focused on five polymorphisms in three genes (HSPA1A, HSPA1B, and HSPA1L) of Hsp70 family to explore the genetic contribution, alone and in combination, of these polymorphisms to essential hypertension risk in a Uygur population. Genotyping was performed using PCR-RFLP and direct sequencing techniques. Data were analyzed using haplotype and multifactor dimensionality reduction (MDR) methods. Genotype distributions of all the polymorphisms satisfied the Hardy-Weinberg proportions in cases and controls. Statistical significance was only observed in the genotype (P = 0.0028) and (P = 0.0146) allele distributions of -110A/C polymorphism, with the -110C allele conferring a 1.45- and 2.83-fold of relative risk, assuming the additive and recessive models, respectively, and in 1267A/G genotype distribution (P = 0.0106) with the 1267G allele conferring a 44% reduced risk. The interaction information analysis indicated that polymorphisms -110A/C and 1267A/G had a strong synergistic effect, while polymorphisms 2074G/C and 2437T/C had a moderate synergistic effect. Haplotype analyses further strengthened the interaction information. Using the haplotype H(1) as a reference, haplotype H(4) had a 40% reduced risk, while haplotypes H(5) and H(8) had a significantly 5.00- and 3.75-fold increased risk for essential hypertension, respectively. Taken together, our results supported strong genetic interaction of the studied polymorphisms with the risk of having essential hypertension in Uygur ethnicity. Functional studies are warranted to confirm or refute these findings. This is the first study to evaluate the genetic interaction information of the Hsp70 in Uygur ethnicity, which represents one of the major nationalities in China with high homogeneity and unique lifestyles. Moreover, we employed the haplotype and MDR methods to explore the potential interaction of Hsp70 genetic polymorphisms in the pathogenesis of essential hypertension in Uygur.

Hypertrophic cardiomyopathy (HCM) is a cardiovascular disease with high heterogeneity. Limited knowledge concerning the genetic background of nearly 40% HCM cases indicates there is a clear need for further investigation to explore the genetic pathogenesis of the disease. In this study, we undertook a whole exome sequencing (WES) approach to identify novel candidate genes and mutations associated with HCM. The cohort consisted of 74 unrelated patients with sporadic HCM (sHCM) previously determined to be negative for mutations in eight sarcomere genes. The results showed that 7 of 74 patients (9.5%) had damaging mutations in 43 known HCM disease genes. Furthermore, after analysis combining the Transmission and De novo Association (TADA) program and the ToppGene program, 10 putative genes gained priority. A thorough review of public databases and related literature revealed that there is strong supporting evidence for most of the genes playing roles in various aspects of heart development. Findings from recent studies suggest that the putative and known disease genes converge on three functional pathways: sarcomere function, calcium signaling and metabolism pathway. This study illustrates the benefit of WES, in combination with rare variant analysis tools, in providing valuable insight into the genetic etiology of a heterogeneous sporadic disease.

Dilated cardiomyopathy (DCM) is a myocardial disease with a high mortality rate. Approximately 40 genes have been found to be associated with DCM to date. Non-familial DCM can also be caused by gene mutations, suggesting that genetic factors were involved in the pathogenesis of DCM; therefore genetic testing is beneficial for the early diagnosis of DCM, which can facilitate the implementation of preventive measures by and within patient's families. Here, we investigated the underlying genetic mutations involved in the cause of patients with DCM. This prospective study included 240 patients with idiopathic DCM and 240 healthy volunteers. Subject clinical data were collected and polymerase chain reaction amplification was carried out on subject DNA for three candidate genes tropomyosin (TPM1), cardiac troponin T type-2 (TNNT2), and nuclear lamina protein A/C. Single nucleotide polymorphism (SNP) loci were detected in the TPM1 (rs1071646) and TNNT2 (rs3729547) genes, respectively. The genotype distributions and allele frequencies were found to satisfy Hardy-Weinberg equilibrium, which indicated that the group was representative. Statistically significant differences were found between the variant frequencies in the two SNP loci between the Kazakh patients with idiopathic DCM (IDCM) and healthy volunteers. A significant difference in the genotype distributions (P = 0.000) and allele frequencies (P = 0.000) of SNP rs1071646, and another significant difference in the genotype distributions (P = 0.000) and allele frequencies (P = 0.039) of SNP rs3729547 between Kazakhs with IDCM and Kazakh controls. These results suggest that the TPM1 (rs1071646) and TNNT2 (rs3729547) gene variants might represent risk factors for patients with DCM in the Kazakh population.

In the present study, the culture conditions for the accumulation of Glycyrrhiza uralensis adventitious root metabolites in balloon-type bubble bioreactors (BTBBs) have been optimized. The results of the culture showed that the best culture conditions were a cone angle of 90° bioreactor and 0.4-0.6-0.4-vvm aeration volume. Aspergillus niger can be used as a fungal elicitor to enhance the production of defense compounds in plants. With the addition of a fungal elicitor (derived from Aspergillus niger), the maximum accumulation of total flavonoids (16.12 mg g(-1)) and glycyrrhetinic acid (0.18 mg g(-1)) occurred at a dose of 400 mg L(-1) of Aspergillus niger resulting in a 3.47-fold and 1.8-fold increase over control roots. However, the highest concentration of polysaccharide (106.06 mg g(-1)) was achieved with a mixture of elicitors (Aspergillus niger and salicylic acid) added to the medium, resulting in a 1.09-fold increase over Aspergillus niger treatment alone. Electrospray ionization tandem mass spectrometry (ESI-MS(n)) analysis was performed, showing that seven compounds were present after treatment with the elicitors, including uralsaponin B, licorice saponin B2, liquiritin, and (3R)-vestitol, only identified in the mixed elicitor treatment group. It has also been found that elicitors (Aspergillus niger and salicylic acid) significantly upregulated the expression of the cinnamate 4-hydroxylase (C4H), β-amyrin synthase (β-AS), squalene epoxidase (SE) and a cytochrome P450 monooxygenase (CYP72A154) genes, which are involved in the biosynthesis of bioactive compounds, and increased superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD) activity.