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Hypertension is the most prevalent chronic disease and a risk factor for various diseases. Although its mechanisms and therapies are constantly being updated and developed, they are still not fully clarified. In recent years, novel gut microbiota and its metabolites have attracted widespread attention. It is strongly linked with physiological and pathological systems, especially TMA and TMAO. TMA is formed by intestinal microbial metabolism of choline and L-carnitine and converted into TMAO by FMO3. This paper collected and collated the latest researches and mainly discussed the following four parts. It introduced gut microbiota; provided a focus on TMA, TMA-producing bacteria, and TMAO; summarized the alternations in hypertensive patients and animals; discussed the mechanisms of TMAO with two respects; and summarized the regulatory factors may be as new interventions and therapies of hypertension. And, more relevant studies are still prospected to be accomplished between hypertension and TMA/TMAO for further clinical services.

Background Based on amyloid cascade and tau hypotheses, protein biomarkers of different Aβ and tau species in cerebrospinal fluid (CSF) and blood/plasma/serum have been examined to correlate with brain pathology. Recently, unbiased proteomic profiling of these human samples has been initiated to identify a large number of novel AD biomarker candidates, but it is challenging to define reliable candidates for subsequent large-scale validation. Methods We present a comprehensive strategy to identify biomarker candidates of high confidence by integrating multiple proteomes in AD, including cortex, CSF and serum. The proteomes were analyzed by the multiplexed tandem-mass-tag (TMT) method, extensive liquid chromatography (LC) fractionation and high-resolution tandem mass spectrometry (MS/MS) for ultra-deep coverage. A systems biology approach was used to prioritize the most promising AD signature proteins from all proteomic datasets. Finally, candidate biomarkers identified by the MS discovery were validated by the enzyme-linked immunosorbent (ELISA) and TOMAHAQ targeted MS assays. Results We quantified 13,833, 5941, and 4826 proteins from human cortex, CSF and serum, respectively. Compared to other studies, we analyzed a total of 10 proteomic datasets, covering 17,541 proteins (13,216 genes) in 365 AD, mild cognitive impairment (MCI) and control cases. Our ultra-deep CSF profiling of 20 cases uncovered the majority of previously reported AD biomarker candidates, most of which, however, displayed no statistical significance except SMOC1 and TGFB2. Interestingly, the AD CSF showed evident decrease of a large number of mitochondria proteins that were only detectable in our ultra-deep analysis. Further integration of 4 cortex and 4 CSF cohort proteomes highlighted 6 CSF biomarkers (SMOC1, C1QTNF5, OLFML3, SLIT2, SPON1, and GPNMB) that were consistently identified in at least 2 independent datasets. We also profiled CSF in the 5xFAD mouse model to validate amyloidosis-induced changes, and found consistent mitochondrial decreases (SOD2, PRDX3, ALDH6A1, ETFB, HADHA, and CYB5R3) in both human and mouse samples. In addition, comparison of cortex and serum led to an AD-correlated protein panel of CTHRC1, GFAP and OLFM3. In summary, 37 proteins emerged as potential AD signatures across cortex, CSF and serum, and strikingly, 59% of these were mitochondria proteins, emphasizing mitochondrial dysfunction in AD. Selected biomarker candidates were further validated by ELISA and TOMAHAQ assays. Finally, we prioritized the most promising AD signature proteins including SMOC1, TAU, GFAP, SUCLG2, PRDX3, and NTN1 by integrating all proteomic datasets. Conclusions Our results demonstrate that novel AD biomarker candidates are identified and confirmed by proteomic studies of brain tissue and biofluids, providing a rich resource for large-scale biomarker validation for the AD community.

The complete chloroplast (cp) genome of Crassocephalum crepidioides was sequenced and assembled for the first time. In this study, the total genome size is 150,596 bp in length and demonstrates a typical quadripartite structure containing a large single copy (LSC, 82,575 bp) and a small single copy (SSC, 18,293 bp), separated by a pair of inverted repeats (IRa, IRb) of 24,864 bp. The G + C content of this cp genome was 37.21%. Gene annotation analysis identified 130 genes including 85 protein-coding genes, 37 transfer RNA, and 8 ribosomal RNA genes. The maximum-likelihood phylogenetic analysis result showed that C. crepidioides was closely related to Nannoglottis ravida in the phylogenetic relationship.

Vinylpyrazine compounds are widely present in foods, especially in hot-processed foods, as a class of flavor compounds; however, their formation mechanism in food systems is still unclear. Therefore, in this study, 2-vinylpyrazine and 2-vinyl-6-methylpyrazine were identified in the Maillard model reaction of D-glucose and glycine. The Maillard model reaction of glucose–glycine was constructed to explore the effects of reaction parameters on vinylpyrazines and the related products. The Maillard reaction of [U-13C6] glucose and glycine was established, and alkylpyrazines and formaldehyde were determined via isotope tracing technique as the precursors of vinylpyrazines. The formation of vinylpyrazines was verified by building a model reaction between alkylpyrazines and formaldehyde. The H/D exchange experiment confirmed that the active site of alkylpyrazines was on the methyl group, which was the reaction site for the condensation reaction of alkylpyrazines with formaldehyde. Results suggest that vinylpyrazines are formed by the condensation reaction of alkylpyrazines and formaldehyde.

Background Study shows that signal transducer and activator of transcription 3 (STAT3) can increase the Warburg effect by stimulating hexokinase 2 in breast cancer and upregulate lactate dehydrogenase A and pyruvate dehydrogenase kinase 1 in myeloma. STAT3 and pyruvate kinase M2 (PKM2) can also be activated and enhance the Warburg effect in hepatocellular carcinoma. Precancerous lesions are critical to human and rodent hepatocarcinogenesis. However, the underlying molecular mechanism for the development of liver precancerous lesions remains unknown. We hypothesized that STAT3 promotes the Warburg effect possibly by upregulating p-PKM2 in liver precancerous lesions in rats. Aim To investigate the mechanism of the Warburg effect in liver precancerous lesions in rats. Methods A model of liver precancerous lesions was established by a modified Solt-Farber method. The liver pathological changes were observed by HE staining and immunohistochemistry. The transformation of WB-F344 cells induced with N-methyl-N'-nitro-N-nitrosoguanidine and hydrogen peroxide was evaluated by the soft agar assay and aneuploidy. The levels of glucose and lactate in the tissue and culture medium were detected with a spectrophotometer. The protein levels of glutathione S-transferase-π, proliferating cell nuclear antigen (PCNA), STAT3, and PKM2 were examined by Western blot and immunofluorescence. Results We found that the Warburg effect was increased in liver precancerous lesions in rats. PKM2 and p-STAT3 were upregulated in activated oval cells in liver precancerous lesions in rats. The Warburg effect, p-PKM2, and p-STAT3 expression were also increased in transformed WB-F344 cells. STAT3 activation promoted the clonal formation rate, aneuploidy, alpha-fetoprotein expression, PCNA expression, G1/S phase transition, the Warburg effect, PKM2 phosphorylation, and nuclear translocation in transformed WB-F344 cells. Moreover, the Warburg effect was inhibited by stattic, a specific inhibitor of STAT3, and further reduced in transformed WB-F344 cells after the intervention for PKM2. Conclusion The Warburg effect is initiated in liver precancerous lesions in rats. STAT3 activation promotes the Warburg effect by enhancing the phosphorylation of PKM2 in transformed WB-F344 cells.

The complete chloroplast (cp) genome of Ehrharta erecta was sequenced and assembled for the first time. In this study, The total genome size is 134,511 bp in length and demonstrates a typical quadripartite structure containing a large single copy (LSC, 95,227 bp) and a small single copy (SSC, 12,306 bp), separated by a pair of inverted repeats (IRa, IRb) of 13,489 bp. The G + C content of this chloroplast genome was 38.76%. Gene annotation analysis identified 130 genes including 84 protein-coding genes, 38 transfer RNA, and 8 ribosomal RNA genes. The maximum-likelihood phylogenetic analysis result showed that E. erecta was closely related to O. sativa in the phylogenetic relationship.

Indole-3-butyric acid (IBA) is widely used to encourage root development in cuttings of general field crops, vegetables, forest trees, fruit trees, and flowers. However, previous studies reported that IBA inhibited the germination of citrus buds via an unknown molecular mechanism. This study aimed to unravel the regulatory mechanisms underlying this inhibition. Citrus apical buds were sprayed with 100 mg ⋅ L−1 IBA. Subsequently, the plant hormone levels were analyzed, and transcriptomic analysis was performed. The results identified 3325 upregulated genes and 2926 downregulated genes in the citrus apical buds. The gene set enrichment analysis method was used to determine the Gene Ontology related to the treatment. Genes were enriched into 157 sets, including 17 upregulated sets and 140 downregulated sets, after indole butyric acid treatment. The upregulated gene sets were related to glucose import, sugar transmembrane transporter activity, and photosynthesis. The downregulated genes were mainly related to the ribosomal subunit and cell cycle process under butyric acid treatment. Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis revealed the enrichment of 11 pathways. Of note, genes related to the ribosome and proteasome pathways were significantly downregulated. Only one pathway was significantly upregulated: the autophagy pathway. Overall, these results provided insights into the molecular mechanisms underpinning the IBA-mediated inhibition of citrus bud germination inhibition. Also, the study provided a large transcriptomics dataset that could be used for further research.

2-Acetylthiazole possesses a nutty, cereal-like and popcorn-like aroma and a low odor threshold, and this compound has been identified in some processed foods, while the formation pathway of 2-acetylthiazole has not been clearly elucidated. Here, a model reaction of d-glucose and l-cysteine was constructed to investigate the formation pathway of 2-acetylthiazole. l-Cysteine, d-glucose and the corresponding intermediates, namely, dicarbonyl compounds (DCs), were involved in the formation of 2-acetylthiazole and detected by high-performance liquid chromatography with tandem mass spectrometry (HPLC-MS/MS), high-performance ion chromatography (HPIC) and HPLC, respectively. The carbon module labeling (CAMOLA) technique revealed that the C-4 and C-5 of 2-acetylthiazole were derived from the carbons of glucose. The potential of glyoxal, which is degraded by glucose, to form 2-acetylthiazole was revealed for the first time. A novel route to form 2-acetylthiazole by the reaction of glyoxal and methylglyoxal produced by d-glucose with H2S and NH3 produced by l-cysteine was proposed.

Soil salinity is a serious abiotic stress worldwide. Pecan plants (Carya illinoensis K. Koch) have been suggested for cultivation in soils with high levels of salinity owing to their huge demand. To understand the potential molecular mechanisms in pecan in response to salt stress, RNA-sequencing technology was used to compare the transcriptomes of pecan plants treated with 0, 0.3 %, or 0.6 % NaCl solutions. The results indicated that 170,086 unigenes were obtained from pecan leaf cDNA samples. Based on the assembled de novo transcriptome, 53, 535, and 7358 differentially expressed genes (DEGs) were detected between untreated and salt-treated leaves at 8, 24, and 48 h, respectively. Because of the large number of DEGs across different contrasts, a Gene Set Enrichment Analysis was selected to identify gene pathways associated with salt treatment. A total of 1858 DEGs were enriched in 66 gene sets, including 22 up-regulated and 47 down-regulated gene sets in the salt treatment groups, compared with those in the control groups. The up-regulated gene sets were mainly involved in the response to salicylic acid; the regulation of the jasmonic acid-mediated signalling pathway during the short-term treatment (8 h); and the cellular response to hypoxia, cellular respiration, and RNA modification during the long-term treatment (24-48 h). The down-regulated gene sets were predominately associated with photosynthesis, water transport, and the metabolic biosynthetic process under salt stress. Genes related to the Really Interesting New Gene superfamily protein and F-box domain protein in the ubiquitin-dependent degradation pathway were significantly up-regulated or down-regulated in different periods of the regulating process. Overall, these results not only enrich genomic resources but also provide insights into the molecular mechanism in pecan under salt stress.

Red bayberry (Morella rubra) is an evergreen fruit tree found in southern China whose whole-genome sequence has recently been published. We updated the linkage map of the species by adding 118 SSR markers and the female-specific marker MrFT2_BD-SEX. The integrated map included eight linkage groups and spanned 491 cM. Eleven sex-associated markers were identified, six of which were located in linkage group 8, in agreement with the previously reported location of the sex-determining region. The MrFT2_BD-SEX marker was genotyped in 203 cultivated accessions. Among the females of the accessions, we found two female-specific alleles, designated W-b (151 bp) and W-d (129 bp). We previously found that‘Dongkui’, a female cultivar, could produce viable pollen (we refer to such plants‘Dongkui-male’) and serve as the paternal parent in crosses. The genotypes of the MrFT2_BD-SEX marker were W-b/Z in‘Biqi’and W-d/Z in‘Dongkui-male’. The progeny of a cross between these parents produced a 3:1 female (W-) to male (ZZ) ratio and the expected 1:1:1:1 ratio of W-b/W-d: W-b/Z: W-d/Z: Z/Z. In addition, the flowering and fruiting phenotypes of all the F1 progeny fit their genotypes. Our results confirm the existence of ZW sex determination and show that the female phenotype is controlled by a single dominant locus (W) in a small genomic region (59 kb and less than 3.3 cM). Furthermore, we have produced a homozygous “super female” (WW) that should produce all-female offspring in the F2 generation, providing a foundation for commercial use and presenting great potential for use in modern breeding programs.

l -asparaginase (EC 3.5.1.1) needs to endure high temperatures during applications in food processing. We here improved the thermal stability of Bacillus subtilis 168 l -asparaginase by regulating both the kinetic and thermodynamic states of the enzyme. According to the molecular dynamics simulation for l -asparaginase, 10 flexible residues potentially affecting the kinetic stability were individually subjected to saturation mutagenesis, generating two highly stabilized mutants, S180N and D289T. As predicted by Discovery Studio, eight residues of l -asparaginase formed multiple unfavourable charge–charge interactions adversely affecting thermodynamic stability. Single-site saturation mutagenesis on these residues resulted in two thermodynamically stabilized mutants, E260F and E292S. Using these mutants, we constructed a further stabilized quadruple mutant NTFS (S180N/D289T/E260F/E292S). Compared with the wild-type enzyme, NTFS exhibited an 8.1-fold increase in half-life at 65 °C and a 5.56 °C increase in melting temperature. NTFS also showed a substantial increase in the transition state energy barrier (ΔΔGtransition = 5.27 KJ/mol) and a clear decrease in folding free energy (ΔΔGfolding = −19.12 kcal/mol) relative to the wild-type enzyme. These results indicated that simultaneous regulation of both the kinetic and thermodynamic states of an enzyme represents an efficient strategy for enhancing thermal stability. The highly stable NTFS described here might be beneficial for industrial application.

In this study, the complete chloroplast (cp) genome of Duranta erecta was assembled using Illumina sequencing data. The complete cp genome is 149,869 bp in length, including a pair of invert repeats (IRA and IRB) regions of 22,839 bp, large single-copy (LSC) region of 86,201 bp, and small single-copy (SSC) region of 17,990 bp. The G + C content of this cp genome was 38.26%. A total of 128 genes were predicted in the genome, including 83 protein-coding genes, 37 tRNA genes, and eight rRNA genes. Phylogenetic analysis confirmed the phylogenetic relationship between D. erecta and other representative species of Verbenaceae.

In this study, the complete chloroplast genome of O. sativa Temperate Japonica YunJing-24 was assembled using Illumina sequencing data. The complete chloroplast (cp) genome is 134,556 bp in length, including a pair of invert repeats (IRA and IRB) regions of 20,797 bp, large single-copy (LSC) region of 80,615 bp, and small single-copy (SSC) region of 12,347 bp. A total of 129 genes were predicted in the genome, including 87 protein-coding genes, 37 tRNA genes, and 8 rRNA genes. Phylogenetic analysis confirmed the phylogenetic relationship between O. sativa Temperate Japonica and other representative species.

Engineering cell factories for producing biofuels and pharmaceuticals has spurred great interests to develop rapid and efficient synthetic biology tools customized for modular pathway engineering. Along the way, combinatorial gene expression control through modification of regulatory element offered tremendous opportunity for fine-tuning gene expression and generating digital-like genetic circuits. In this report, we present an efficient evolutionary approach to build a range of regulatory control elements. The reported method allows for rapid construction of promoter, 5′UTR, terminator and trans-activating RNA libraries. Synthetic overlapping oligos with high portion of degenerate nucleotides flanking the regulatory element could be efficiently assembled to a vector expressing fluorescence reporter. This approach combines high mutation rate of the synthetic DNA with the high assembly efficiency of Gibson Mix. Our constructed library demonstrates broad range of transcriptional or translational gene expression dynamics. Specifically, both the promoter library and 5′UTR library exhibits gene expression dynamics spanning across three order of magnitude. The terminator library and trans-activating RNA library displays relatively narrowed gene expression pattern. The reported study provides a versatile toolbox for rapidly constructing a large family of prokaryotic regulatory elements. These libraries also facilitate the implementation of combinatorial pathway engineering principles and the engineering of more efficient microbial cell factory for various biomanufacturing applications.

A novel Gram-staining positive, moderately halophilic, endospore-forming, motile, rod-shaped and strictly aerobic strain, designated YIM 93565T, was isolated from a salt lake in Xinjiang province of China and subjected to a polyphasic taxonomic study. Strain YIM 93565T grew in the range of pH 6.0–9.0 (optimum pH 7.0), 10–45 °C (optimum 35–40 °C) and at salinities of 2–24% (w/v) NaCl (optimum 7–10%). Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain YIM 93565T clustered with members of the genera Gracilibacillus and form a clade with Gracilibacillus bigeumensis KCTC 13130T (95.6% similarity) and Gracilibacillus halophilus DSM 17856T (94.9%), which was well separated from others. The DNA G + C content of this novel strain was 36.8 mol%. The major fatty acids were anteiso-C15:0, iso-C15:0, C16:0 and anteiso-C17:0 and its polar lipids consisted of diphosphatidylglycerol, phosphatidylglycerol, one unidentified glycolipid and two unidentified phospholipids. The predominant menaquinone was MK-7. The cell-wall peptidoglycan was based on meso-diaminopimelic acid. Based on the results of phylogenetic, physiological and chemotaxonomic comparative analyses, the isolate is assigned to a novel species of the genus Gracilibacillus, for which the name Gracilibacillus eburneus sp. nov. is proposed, with the type strain YIM 93565T (= DSM 23710T = CCTCC AB 2013249T).

Atherosclerosis (AS) is a progressive disease of multifactorial origin, which occurs in response to endothelial injury. Increased homocysteine (Hcy) is considered a major cause of endothelial dysfunction, oxidative stress and DNA methylation; however, the mechanisms remain to be fully elucidated. The aim of the present study was to investigate whether Hcy causes injury to endothelial cells (ECs) by the effect of lectin‑like oxidized‑low density lipoprotein receptor‑1 (LOX‑1) DNA methylation through toll‑like receptor 4(TLR4)/nuclear factor (NF)‑κB/DNA methyltransferase (DNMT)1. The ECs were treated with different concentrations of Hcy, and it was found that Hcy promoted the expression of TLR4, leading to EC injury. The effect of oxidative stress was analyzed by measuring superoxide dismutase, malondialdehyde and hydrogen peroxide in the ECs. In addition, the association between NF‑κB and DNMT1 was examined by treatment of the ECs with pyrrolidine dithiocarbamate (PDTC). The results suggested that Hcy induced LOX‑1 DNA hypomethyaltion to promote the expression levels of LOX‑1. Taken together, Hcy injured the ECs through the effect of methylation and trans‑sulfuration metabolism of LOX‑1 through TLR4/NF‑κB/DNMT1. Following injury to the ECs, lipids, particularly ox‑LDL, accumulated in the sub‑endothelial layer to promote the formation of AS.

Alzheimer's disease (AD) is one of the most common neurodegenerative diseases. The formation of amyloid plaques by aggregated amyloid beta (Aβ) peptides is a primary event in AD pathology. Understanding the metabolomic features and related pathways is critical for studying plaque-related pathological events (e.g., cell death and neuron dysfunction). Mass spectrometry imaging (MSI), due to its high sensitivity and ability to obtain the spatial distribution of metabolites, has been applied to AD studies. However, limited studies of metabolites in amyloid plaques have been performed due to the drawbacks of the commonly used techniques such as matrix-assisted laser desorption/ionization MSI. In the current study, we obtained high spatial resolution (∼17 μm) MS images of the AD mouse brain using the Single-probe, a microscale sampling and ionization device, coupled to a mass spectrometer under ambient conditions. The adjacent slices were used to obtain fluorescence microscopy images to locate amyloid plaques. The MS image and the fluorescence microscopy image were fused to spatially correlate histological protein hallmarks with metabolomic features. The fused images produced significantly improved spatial resolution (∼5 μm), allowing for the determination of fine structures in MS images and metabolomic biomarkers representing amyloid plaques.

The flavor release mechanism related to the interaction of aroma compounds with proteins is still unclear. In this study, the interaction of protein with pyrazine homologues, such as 2-methylpyrazine (MP), 2,5-dimethylpyrazine (DP), 2,3,5-trimethylpyrazine (TRP) and 2,3,5,6-tetramethylpyrazine (TEP), was investigated to elucidate the effect of alkyl distribution in a pyrazine ring on its flavor release in bovine serum albumin (BSA) solution (pH 7.4). The results of SPME-GC-MS indicated that methyl distribution in a pyrazine ring significantly affected its release from BSA solution. The pyrazines released from BSA solution with an increasing order of MP, DP, TRP and TEP. The inhibition mechanism of alkyl-pyrazine release was further elucidated by the interaction between alkyl-pyrazines and BSA using multiple spectroscopic methods. The non-covalent interaction between alkyl-pyrazines and BSA was confirmed as the main interaction force by the value of the bimolecular quenching constant (Kq > 2 × 1010 L mol−1 s−1). A decrease in the hydrophobicity of the microenvironment between the alkyl-pyrazine and BSA was detected by synchronous fluorescence spectra, which revealed that alkyl-pyrazines were mainly bound on the sites of tyrosine and tryptophan in BSA. The UV-vis absorption spectra and circular dichromatic (CD) spectrum revealed that alkyl-pyrazines could induce polarity and conformation change of BSA. The above results indicated that the structure of the flavor homologues can affect their release in food matrices.

Alzheimer's disease (AD) displays a long asymptomatic stage before dementia. We characterize AD stage-associated molecular networks by profiling 14,513 proteins and 34,173 phosphosites in the human brain with mass spectrometry, highlighting 173 protein changes in 17 pathways. The altered proteins are validated in two independent cohorts, showing partial RNA dependency. Comparisons of brain tissue and cerebrospinal fluid proteomes reveal biomarker candidates. Combining with 5xFAD mouse analysis, we determine 15 Aβ-correlated proteins (e.g., MDK, NTN1, SMOC1, SLIT2, and HTRA1). 5xFAD shows a proteomic signature similar to symptomatic AD but exhibits activation of autophagy and interferon response and lacks human-specific deleterious events, such as downregulation of neurotrophic factors and synaptic proteins. Multi-omics integration prioritizes AD-related molecules and pathways, including amyloid cascade, inflammation, complement, WNT signaling, TGF-β and BMP signaling, lipid metabolism, iron homeostasis, and membrane transport. Some Aβ-correlated proteins are colocalized with amyloid plaques. Thus, the multilayer omics approach identifies protein networks during AD progression.

The dispersion solid-phase microextraction (DSPE) combined with dispersion liquid–liquid microextraction (DLLME) was developed as a rapid, simple and effective pretreatment method for the determination of flavor enhancers (maltol, ethyl maltol, vanillin, methyl vanillin, ethyl vanillin) of ready-to-eat seafood products (dried squid, baked squid, fried fish, steamed abalone). Under the optimized DSPE–DLLME method, the developed method exhibited low limits of quantitation (0.20–0.50 μg g−1) and excellent linearity (R2 ≥ 0.995). The recoveries of flavor enhancers in the actual samples were 83.7%–114.9%. Compared with traditional pretreatment methods, the developed DSPE–DLLME method was rapid (17 min) and easy to determine the flavor enhancers by high performance liquid chromatography with photodiode array detector (HPLC-PDA). In the actual samples, creamy compounds such as vanillin, methyl vanillin and ethyl vanillin were hardly found. However, the flavor compounds produced by thermal reactions such as maltol (except for dried squid 3) and ethyl maltol were present in all samples.

LncRNA SBF2-AS1 has been reported to be implicated in the deterioration of multiple human cancers. However, the roles and underlying mechanisms of SBF2-AS1 in acute myeloid leukemia (AML) are still unclear. In the present study, the online GEPIA database showed that SBF2-AS1 expression was significantly increased in AML samples. QRT-PCR results showed that SBF2-AS1 expression was upregulated in AML cells. CCK-8 assay revealed that SBF2-AS1 inhibition decreased AML cells proliferation ability in vitro. Flow cytometry assays showed that SBF2-AS1 inhibition induced AML cells apoptosis and arrested AML cells in G0/G1 phase. Mechanistically, miR-188-5p was identified as a direct target of SBF2-AS1. SBF2-AS1 upregulated the expression level of ZFP91 by sponging miR-188-5p. And the effects of SBF2-AS1 suppression on AML cells progression could be abolished by miR-188-5p inhibitors. Moreover, we found that SBF2-AS1 inhibition reduced tumor growth in vivo. Taken together, our findings elucidated that SBF2-AS1 could act as a miRNA sponge in AML progression, and provided a potential therapeutic strategy for AML treatment.

L-asparaginase (EC 3.5.1.1, ASN) exhibits great commercial value due to its uses in the food and medicine industry. In this study, we reported the enhanced expression of type II ASN from Bacillus subtilis 168 in B. subtilis WB600 through a combined strategy. First, eight signal peptides (the signal peptide of the ASN, ywbN, yvgO, amyE, oppA, vpr, lipA, and wapA) were used for ASN secretion in B. subtilis by using Hpa II promoter, respectively. The signal peptide wapA achieved the highest extracellular ASN activity (28.91 U/mL). Second, Hpa II promoter was replaced by a strong promoter, P43 promoter, resulting in 38.1 % enhanced ASN activity. By two rounds of error-prone PCR mutation, the P43 promoter variants with remarkably enhanced strength (D7, E2, H6, B2, and F3) were identified. B2 (−28: A → G, −13: A → G) achieved ASN activity up to 51.13 U/mL. Third, after deletion of the N-terminal 25-residues, ASN activity reached 102.41 U/mL, which was 100 % higher than that of the intact ASN. At last, the extracellular ASN of the B. subtilis arrived at 407.6 U/mL (2.5 g/L of ASN protein) in a 3-L bioreactor by using a fed-batch strategy. The purified ASN showed maximal activity at 65 °C and its half-life at 65 °C was 61 min. The K m and k cat of the ASN were 5.29 mM and 54.4 s−1, respectively. To the best of our knowledge, we obtained the highest yield of ASN in a food-grade host ever reported, which may benefit the industrial production and application of ASN.

The peach [Prunus persica (L.) Batsch] is an important fruit crop that is prone to injury during spring frost, yet little is known about the molecular mechanism of the low temperature (LT) stress response in the peach. In this study, we employed next generation sequencing-technology to characterize the transcriptome of the peach stigma and identify differentially expressed genes (DEGs) of peaches treated at LT (−2 °C) for 4 and 26 h. We obtained a total of 31,273,923 (CK), 37,477,875 (S1), and 36,009,843 (S2) perfect reads from controls, 4 h LT treatment, and 26 h LT treatment, respectively. 96.4–96.8 % of the reads mapped to the peach genome. Gene ontology and pathway enrichment analysis revealed that the upregulated DEGs were predominately involved in response to temperature stimulus, carbohydrate metabolism, biological processes, cytoskeletal, hydrolase activity and primary metabolic pathways. Many of the genes coded for proteins involved in response to LT-associated mechanisms, including the U-box domain-containing protein, CBL-interacting protein kinase, NAC domain-containing protein, and dehydration-responsive element-binding protein. More genes were differentially expressed in S2 (966) compared to S1 (276), and of these, the U-box domain-containing protein 34 had the highest expression in S2. Furthermore, we confirmed differential expression of 20 selected genes via qPCR and found highly similar expression differences to the Illumina analysis. Overall, we identified candidate genes for LT stress in the peach and provide new insights for investigating the molecular mechanisms of LT stress.

Hyperhomocysteinemia (HHcy) is an independent risk factor for the development of atherosclerosis (AS), according to overwhelming number of clinical and epidemiological studies. However, the underlying pathogenic molecular mechanisms by which HHcy promotes AS remain to be fully elucidated. Fatty acid binding protein 4 (FABP4) has been shown to be important in macrophage cholesterol trafficking. The objective of the present study was to determine whether homocysteine (Hcy) accelerates AS through regulating FABP4, and then mediates cholesterol accumulation in macrophages. Hcy concentrations of 0, 50, 100, 200 and 500 µM, and 100 µM Hcy+30 µM vitamin B12 (VB12)+30 µM folic acid (FA) were respectively added to cultured THP‑1 monocyte‑derived macrophages for 24 h. The levels of FABP4, which acts as a key factor connecting cellular lipid accumulation to inflammation, were determined using reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) and western blot analyses in the macrophages. The present study used a nested touchdown methylation‑specific PCR assay to detect the DNA methylation status of the FABP4 promoter region. In addition, the FABP4 gene fragment was inserted into the cloning vector, pcDNA3.1‑EGFP, to construct the recombinant plasmid, pcDNA3.1‑EGFP/FABP4, which was identified using restriction endonuclease digestion analysis and DNA sequencing. The pcDNA3.1‑EGFP/FABP4 expression plasmid was transfected into THP‑1 monocyte‑derived macrophages, mediated by liposome reagent, following which the expression levels of FABP4 were detected using RT‑qPCR and western blot analyses. The present study also determined the intracellular accumulation of total cholesterol in the macrophages. The results indicated that Hcy decreased the levels of FABP4 promoter methylation, but increased the mRNA and protein expression levels of FABP4 in the macrophages, compared with the control group (0 µM Hcy). However, no dose‑dependent changes were observed with increasing concentrations of Hcy. The recombinant fluorescent eukaryotic expression vector, pcDNA3.1‑EGFP/FABP4, was successfully constructed and effectively expressed in the THP‑1 macrophages. The results also showed that FABP4 accelerated the accumulation of cholesterol in the macrophages. Taken together, the results of the present study suggested that FABP4 DNA hypomethylation induced by Hcy may be involved in the overexpression of FABP4, thereby inducing cholesterol accumulation in macrophages.

Dendrobium candidum Lindl. is an endangered lithophytic orchid distrubuted in the subtropical regions of China which was listed in the category of the Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES). Here, we reported the complete chloroplast (cp) genome sequence and the cp genomic features of D. candidum. The genome was 152,094 bp long with 129 genes comprising 83 protein-coding genes, 40 tRNA genes, and six rRNA genes. Most of the genes occurred as a single-copy, while 18 occurred in two copies. Phylogenetic analysis of a data set of cp genomes indicated that D. candidum is closely related to that of other species in Dendrobium.

Dried Lentinus edodes (Berk.) sing stipe (LES) is mostly discarded owing to its high fibre content and bad palatability, causing significant waste of resources. The aim of this study is to sustainably use LES for the production of flavour soybean oil using the supercritical CO2 fluid extraction (SFE) method. The quality properties of the obtained soybean oil flavoured by LES (OFLS) were evaluated in this study. Profiles obtained with an electronic nose and gas chromatography results obtained by quadrupole time-of-flight mass spectrometry and with a mass-selective detector indicated that mushroom-like aroma compounds were mainly concentrated into the OFLSs from separation kettle A (9 MPa). Interestingly, off-flavour compounds from soybean oil, such as o-xylene and trans-2-nonenal, were mostly distilled into separation kettle B (4 MPa). More importantly, the developed SFE method effectively improved the colour quality of the OFLSs. Additionally, the decrease of the viscosity of the OFLSs indicated that the thermal stability of the edible oil could be improved. In summary, the OFLSs prepared by the green and sustainable SFE method significantly improved the quality of the edible oil.

l -Asparaginase (ASN; EC 3.5.1.1) shows great commercial value because of its ability to reduce toxic levels of acrylamide in foods. To achieve high-efficiency production of l -asparaginase, an open reading frame of 978 bp encoding asparaginase (BcA) was amplified from Bacillus cereus BDRD-ST26, followed by its expression in Bacillus subtilis WB600, with the highest yield of 374.9 U/ml obtained using an amyE-signal peptide. A four-step purification protocol was used to purify BcA, resulting in a 15.1-fold increase in purification yield, with a specific activity of purified BcA at 550.8 U/mg and accompanied by detection of minimal l -glutaminase activity. Maximum BcA activity was detected at 50°C and pH 9.0 in 20 mM Tris–HCl buffer, with a half-life at 50°C of 17.35 min and a Km and kcat of 9.38 mM and 63.6 s−1, respectively. Compared with untreated potato strips, 72% acrylamide (2.35 mg/kg) was removed from potato strips pretreated with BcA. These results indicated that this novel BcA variant represents a potential candidate for application in the food-processing industry.

Previous brain morphology-related diagnostic models for attention-deficit hyperactivity disorder (ADHD) were based on regional features. However, building a model of individual interregional morphological connectivity is a challenging task. This study aimed to identify children with ADHD utilizing a novel interregional morphological connectivity model and discover the discriminative patterns in patients. Therefore, novel interregional morphological patterns rather than regional patterns were extracted via surface-based analysis. The interregional morphological features were trained and tested using a hybrid machine learning method, which was implemented using the leave-one-out cross-validation (LOOCV) method to produce the optimized discriminative model and discriminative patterns. The inclusion of interregional morphological connectivity significantly improved the performance of the diagnostic models compared to the performance of the model constructed using regional features. The optimized discriminative model exhibited a total accuracy of 74.65%, a sensitivity of 75% and a specificity of 74.29%. The brain regions displaying altered morphological connectivity included the insula, the caudal anterior cingulate cortex, the frontal pole, and the postcentral cortex, among others. In addition, the altered connections correlated with the clinical symptoms. In summary, patients with ADHD exhibited altered morphological connectivity, which might be a potential biomarker for the classification of ADHD. The discriminative features will potentially benefit studies investigating the brain network mechanisms of ADHD.

Elevated homocysteine (Hcy) levels have been reported to be involved in liver injury, and autophagy plays an important role in normal hepatic physiology and pathophysiology, but the mechanism underlying Hcy regulated autophagy is currently unknown. In this study, CBS+/- mice were fed with regular diet for 12 weeks to establish a hyperhomocysteinemia (HHcy) model and HL-7702 cells were treated with Hcy, we found that Hcy increases autophagy and aggravates liver injury by downregulation of cystic fibrosis transmembrane conductance regulator (CFTR) expression in vivo and in vitro. Overexpression of CFTR inhibited the formation of autophagosomes and the expression of autophagy-related proteins BECN1, LC3-II/I and Atg12, while the expression of p62 increased in Hcy-treated hepatocytes and CBS+/- mice injected with lentivirus expressing CFTR. Further study showed that CFTR expression is regulated by the interaction of DNA methyltransferase 1 (DNMT1) and enhancer of zeste homolog 2 (EZH2), which, respectively, regulate DNA methylation and histone H3 lysine 27 trimethylation (H3K27me3). In conclusion, our study showed that Hcy activates autophagy by inhibition of CFTR expression via interaction between H3K27me3 and DNA methylation in the mouse liver. These findings provide new insight into the mechanism of Hcy-induced autophagy in liver injury.

Exercise reduces the risk of developing a number of neurological disorders and increases the efficiency of cellular energy production. However, overly strenuous exercise produces oxidative stress. Proper oxygenation is crucial for the health of all tissues, and tight regulation of cellular oxygen is critical to balance O2 levels and redox homeostasis in the brain. Hypoxia Inducible Factor (HIF)1α and HIF2α are transcription factors regulated by cellular oxygen concentration that initiate gene regulation of vascular development, redox homeostasis, and cell cycle control. HIF1α and HIF2α contribute to important adaptive mechanisms that occur when oxygen and ROS homeostasis become unbalanced. It has been shown that preconditioning by exposure to a stressor prior to a hypoxic event reduces damage that would otherwise occur. Previously we reported that 3months of exercise protects SNpc dopaminergic (DA) neurons from toxicity caused by Complex I inhibition. Here, we identify the cells in the SNpc that express HIF1α and HIF2α and show that running exercise produces hypoxia in SNpc DA neurons, and alters the expression of HIF1α and HIF2α. In mice carrying a conditional knockout of Hif1α in postnatal neurons we observe that exercise alone produces SNpc TH+ DA neuron loss. Loss of HIF1α also abolishes exercise-induced neuroprotection. In mice lacking Hif2α in postnatal neurons, the number of TH+ DA neurons in the adult SNpc is diminished, but 3months of exercise rescues this loss. We conclude that HIF1α is necessary for exercise-induced neuroprotection and both HIF1α and HIF2α are necessary for the survival and function of adult SNpc DA neurons.

An earlier molecular phylogenetic analysis using cpDNA sequence data revealed the non-monophyly of Primula subgenera Auganthus and Carolinella, and the evolutionary implication of the infrageneric division of these two groups remains uncertain. In the present study, 130 new sequences were newly analyzed, with the combination of plastid genes and nrDNA ITS regions through maximum parsimony and Bayesian inference analyses. It confirms that neither subgenus Auganthus nor subgenus Carolinella is monophyletic in its current circumscription. The calyptrate capsule, a diagnostic character for subgenus Carolinella, may have evolved multiple times in Primula. Subgenus Auganthus would be a natural group only with inclusion of subgenus Carolinella and exclusion of P. aromatica. The positions of some infrageneric groups are clarified, but challenges remain regarding relevant morphological characters for sectional delimitations within subgenus Auganthus.

Morella rubra, red bayberry, is an economically important fruit tree in south China. Here, we assembled the first high‐quality genome for both a female and a male individual of red bayberry. The genome size was 313‐Mb, and 90% sequences were assembled into eight pseudo chromosome molecules, with 32 493 predicted genes. By whole‐genome comparison between the female and male and association analysis with sequences of bulked and individual DNA samples from female and male, a 59‐Kb region determining female was identified and located on distal end of pseudochromosome 8, which contains abundant transposable element and seven putative genes, four of them are related to sex floral development. This 59‐Kb female‐specific region was likely to be derived from duplication and rearrangement of paralogous genes and retained non‐recombinant in the female‐specific region. Sex‐specific molecular markers developed from candidate genes co‐segregated with sex in a genetically diverse female and male germplasm. We propose sex determination follow the ZW model of female heterogamety. The genome sequence of red bayberry provides a valuable resource for plant sex chromosome evolution and also provides important insights for molecular biology, genetics and modern breeding in Myricaceae family.

Missense mutations in the leucine rich repeat kinase 2 (LRRK2) gene result in late-onset Parkinson’s disease. The incomplete penetrance of LRRK2 mutations in humans and LRRK2 murine models of Parkinson’s disease suggests that the disease may result from a complex interplay of genetic predispositions and persistent exogenous insults. Since neuroinflammation is commonly associated with the pathogenesis of Parkinson’s disease, we examine a potential role of mutant LRRK2 in regulation of the immune response and inflammatory signalling in vivo. Here, we show that mice overexpressing human pathogenic LRRK2 mutations, but not wild-type mice or mice overexpressing human wild-type LRRK2 exhibit long-term lipopolysaccharide-induced nigral neuronal loss. This neurodegeneration is accompanied by an exacerbated neuroinflammation in the brain. The increased immune response in the brain of mutant mice subsequently has an effect on neurons by inducing intraneuronal LRRK2 upregulation. However, the enhanced neuroinflammation is unlikely to be triggered by dysfunctional microglia or infiltrated T cells and/or monocytes, but by peripheral circulating inflammatory molecules. Analysis of cytokine kinetics and inflammatory pathways in the peripheral immune cells demonstrates that LRRK2 mutation alters type II interferon immune response, suggesting that this increased neuroinflammatory response may arise outside the central nervous system. Overall, this study suggests that peripheral immune signalling plays an unexpected—but important—role in the regulation of neurodegeneration in LRRK2-associated Parkinson’s disease, and provides new targets for interfering with the onset and progression of the disease.

In recent years, coinfection of tuberculosis (TB) and parasitosis in humans is an emerging problem in coendemic areas, which has been increasingly highlighted in developing countries. However, there is limited information about the prevalence of Toxoplasma gondii infection in TB patients. Therefore, through a case-control study, 924 TB patients hospitalized for diacrisis or treatment in northeastern and eastern China, and 924 control subjects from the general population of the same region matched with gender, age, and residence were examined for the presence of IgG and IgM antibodies to T. gondii and associated sociodemographic and behavioral characteristics in a population of TB patients. Seroprevalence of IgG antibodies to T. gondii in TB patients (122/924, 13.2%) was significantly higher than control subjects (90/924, 9.7%) (p = 0.019), and 26 (2.8%) TB patients and 19 (2.1%) controls were positive for anti-T. gondii IgM antibodies (p = 0.291), respectively. Multivariate analysis showed that T. gondii infection was associated with keeping cats at home, presence of stray cats, and consumption of raw/undercooked meat. The present study first revealed the seroprevalence of T. gondii infection in TB patients in China. Moreover, parasitological surveys should be regularly carried out among TB patients, aiming to prevent the possibility of severe toxoplasmosis.

Vascular smooth muscle cell (VSMC) proliferation is a primary pathological event in the development of atherosclerosis (AS), and the presence of homocysteine (Hcy) acts as an independent risk factor for AS. However, the underlying mechanisms remain to be elucidated. Phosphatase and tensin homologue on chromosome 10 (PTEN), is endogenously expressed in VSMCs and induces multiple signaling networks involved in cell proliferation, survival and inflammation, however, the specific role of PTEN is still unknown. The present study detected the proliferation ratio of VSMCs following treatment with Hcy and Resveratrol (RSV). In the 100 µM Hcy group, the proliferation ratio increased, and treatment with RSV decreased the proliferation ratio induced by Hcy. Reverse transcription‑quantitative polymerase chain reaction and western blotting were used to analyze PTEN expression, RSV treatment was associated with decreased PTEN expression levels in VSMCs. PTEN levels were decreased in Hcy treated cells, and the proliferation ratio of VSMCs were increased following treated with Hcy. To study the mechanism of regulation of PTEN by Hcy, the present study detected PTEN methylation levels in VSMCs, and PTEN DNA methylation levels were demonstrated to be increased in the 100 µM Hcy group, whereas treatment with RSV decreased the methylation status. DNA methyltransferase 1 is important role in the regulation of PTEN methylation. Overall, Hcy impacts the methylation status of PTEN, which is involved in cell proliferation, and induces the proliferation of VSMCs. This effect is alleviated by treatment with RSV, which exhibits an antagonistic mechanism against Hcy.

One new tirucallane-type nortriterpenoid Nortirucallane A (1), together with Chrysoeriol (2) and Isorhamnetin (3), was isolated from the methylene chloride part of Lonicerae japonicae flos. Their structures were elucidated by the detailed analysis of comprehensive spectroscopic data. Compound 3 was isolated from the genus of Lonicera for the first time. The significance of 1 for the study of phytochemical taxonomy was discussed.

Ipomoea trifida is a herbaceous plant of the family Convolvulaceae that is mainly distributed around the Caribbean Sea. In this study, we reported and analyzed the complete chloroplast genome of I....

Ethnopharmacological relevance Mistletoe ( Viscum coloratum (Kom.) Nakai ) has long been categorized as a traditional herbal medicine in Asia. In addition to its application in cancer therapy, mistletoe has also been used in the treatment of chronic hepatic disorders in China. In the present study, we investigated the antifibrotic effect and mechanisms of action of mistletoe extracts in a rat model of carbon tetrachloride (CCl 4 )-induced hepatotoxicity. Materials and methods An experimental model of hepatic fibrosis was established by intraperitoneal injection of rats with CCl 4 for 8 weeks. Rats were subsequently treated with a mistletoe alkaloid fraction preparation via oral administration (120 mg/kg daily for 8 weeks) or with distilled water as a control. Histopathological changes were observed by hematoxylin and eosin staining and Masson׳s trichrome staining. The expression of markers relevant to hepatic stellate cell (HSC) activation in the liver was assessed by real-time reverse transcription-polymerase chain reaction, immunohistochemistry and western blotting. The anti-fibrosis activity and mechanisms of action of mistletoe alkaloid fractions were further investigated in the HSC-T6 HSC line, following treatment with mistletoe alkaloid fractions (12 mg/ml) for 48 h. Results Hepatic fibrosis decreased markedly in CCl 4 -treated animals following treatment with mistletoe alkaloid fractions , compared to controls. The mRNA levels of transforming growth factor-β1 (TGF-β1), procollagen I and tissue inhibitors of metalloproteinases (TIMPs) were significantly downregulated, by about 40%, 40% and 45%, respectively, in liver tissues from rats treated with mistletoe alkaloid fractions . Furthermore, significant downregulation of TGF-β1, TGF-β1 receptor, phosphorylated Smad 2 and alpha smooth muscle actin (α-SMA) proteins, by about 45%, 30% and 40%, respectively, was also observed in liver tissues from mistletoe alkaloid fractions -treated rats. In contrast, Smad 7 levels were significantly increased by about 30% in mistletoe alkaloid fractions -treated rats. Treatment of HSC-T6 cells with mistletoe alkaloid fractions significantly induced Smad 7 expression and inhibited the expression of α-SMA, TGFβ1, TGF-β1 receptor, Smad 2 and TIMP-1, in vitro. Conclusion We demonstrate that mistletoe alkaloid fractions decrease extracellular matrix accumulation by inhibiting HSC activation. Mechanistically, this may occur via inhibition of TGF-β1/Smad 2 and Smad 7 signal transduction, thereby blocking the synthesis of procollagen I and TIMP-1. These findings suggest that mistletoe alkaloid fractions may be a potential therapeutic agent for the treatment of hepatic fibrosis.

Sequence-specific amplified polymorphism (SSAP) technology is a novel, anchored PCR approach derived from AFLP, which amplifies the region between a transposon insertion and an adjacent restriction site and have higher levels of polymorphism. In the current study, we developed 16 SSAP markers based on the long terminal repeat (LTR) sequences of Ty1- copia retrotransposons in the peach and used them for DNA profiling of 52 individual peaches: 44 peach cultivars and 8 ornamental peaches. These primer combinations produced a total of 1,553 fragments and 1,517 polymorphic bands with a polymorphism percentage of 97.7%. Furthermore, the Shannon's information index of each primer combination ranged from 0.1593 to 0.4456. Neighbor-joining analyses revealed two main genetic clusters, corresponding to the fruit flesh types: (A-1) MF (melting flesh) with clingstone and ornamental peaches; (A-2) MF with freestone and NMF (non-melting flesh) with clingstone. Finally, cluster analyses revealed that all ornamental peaches are closely related to the MF with clingstone peach cultivars. The application of these primer combinations identified using SSAP will facilitate future cultivar identification and germplasm management in peaches.

A Gram-negative, moderately halophilic, strictly aerobic strain, designated YIM 95345T, was isolated from a soil sample of a hypersaline mine in Yunnan province, PR China, and subjected to a polyphasic taxonomic study. Strain YIM 95345T grew at 15–45 °C (optimum 30–35 °C), 3.0–23.0 % (w/v) NaCl (optimum 10.0–11.0 %, w/v) and pH 6.0–9.0 (optimum pH 7.0–8.0). Phylogenetic analyses based on 16S rRNA gene sequences revealed that the organism belongs to the genus Aquisalimonas and exhibited sequence similarity of 96.6 % to the sole type strain Aquisalimonas asiatica CG12T. The predominant isoprenoid quinone was Q-8 and the major fatty acids were C16 : 0, C19 : 0 cyclo ω8c and C18 : 1ω7c. The polar lipids consisted of diphosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylinositol, three aminolipids and three unidentified phospholipids. The G+C content of the genomic DNA was 59.4 mol%. Based on the results of our comparative phylogenetic, chemotaxonomic and physiological analyses, the new isolate is assigned to a novel species of the genus Aquisalimonas , for which the name Aquisalimonas halophila sp. nov. is proposed, with the type strain YIM 95345T ( = DSM 25902T = CCTCC AB 2012043T).

Dendrobium bellatulum Rolfe is an Endangered orchid species that is distributed in the subtropical regions of Yunnan Province, China. It was listed in the category of the Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES). Here, we reported the complete chloroplast (cp) genome sequence and the cp genomic features of D. bellatulum. The genome was 152,107 bp long with 129 genes comprising 83 protein-coding genes, 40 tRNA genes, and 6 rRNA genes. Phylogenetic analysis of a data set of cp genomes indicated that D. bellatulum is clustered with other species in Dendrobium.

Background Casticin, the flavonoid extracted from Vitex rotundifolia L, exerts various biological effects, including anti-inflammatory and anti-cancer activity. The aim of this study is to investigate the effects and mechanisms of casticin in human gallbladder cancer cells. Methods Human NOZ and SGC996 cells were used to perform the experiments. CCK-8 assay and colony formation assay were performed to evaluate cell viability. Cell cycle analyses and annexin V/PI staining assay for apoptosis were measured using flow cytometry. Western blot analysis was used to evaluate the changes in protein expression, and the effect of casticin treatment in vivo was experimented with xenografted tumors. Results In this study, we found that casticin significantly inhibited gallbladder cancer cell proliferation in a dose- and time-dependent manner. Casticin also induced G0/G1 arrest and mitochondrial-related apoptosis by upregulating Bax, cleaved caspase-3, cleaved caspase-9 and cleaved poly ADP-ribose polymerase expression, and by downregulating Bcl-2 expression. Moreover, casticin induced cycle arrest and apoptosis by upregulating p27 and downregulating cyclinD1/cyclin-dependent kinase4 and phosphorylated protein kinase B. In vivo, casticin inhibited tumor growth. Conclusion Casticin induces G0/G1 arrest and apoptosis in gallbladder cancer, suggesting that casticin might represent a novel and effective agent against gallbladder cancer.

Background Epidermal growth factor receptor (Egfr) has been the target of several drugs for cancers. The potential gender differences in genes in the Egfr axis have been suggested in humans and in animal models. Female and male mice from the same recombinant inbred (RI) strain have the same genomic components except the sex difference. A population of different RI mouse strains allows to conduct precise analysis of molecular pathways and regulation of Egfr between female and male mice. Methods The whole genome expression profiles of 70 genetically diverse RI strains of mice were used to compare three major molecular aspects of Egfr gene: the relative expression levels, gene network and expression quantitative trait loci (eQTL) that regulate the expression of Egfr between female and male mice. Results Our data showed that there is a significant sex difference in the expression levels in kidney. A considerable number of genes in the gene network of Egfr are sex differentially expressed. The expression levels of Egfr in mice are statistical significant different between C57BL/6 J (B6) and DBA/2 J (D2) genotypes in male while no difference in female mice. The eQTLs that regulate the expression levels of Egfr between female and male mice are also different. Furthermore, the differential expression levels of Egfr showed significantly different correlations with two known biological traits between male and female mice. Conclusion Overall there is a substantial sex difference in the Egfr pathways in mice. These data may have significant impact on drug target design, development, formulation, and dosage determinant for women and men in clinical trials. Electronic supplementary material The online version of this article (doi:10.1186/s12863-016-0449-3) contains supplementary material, which is available to authorized users.

Vascular smooth muscle cell (VSMC) proliferation is a primary pathological event in atherosclerosis (AS), and homocysteine (Hcy) is an independent risk factor for AS. However, the underlying mechanisms are still lagging. Studies have used the combination of methylation of promoters of multiple genes to diagnose tumors, thus the aim of the current study was to investigate the role of methylation status of several genes in VSMCs treated with Hcy. CpG islands were identified in the promoters of platelet‑derived growth factor (PDGF), p53, phosphatase and tensin homologue on chromosome 10 (PTEN) and mitofusin 2 (MFN2). Hypomethylation was observed to occur in the promoter region of PDGF, hypermethylation in p53, PTEN and MFN2, and hypomethylation in two global methylation indicators, aluminium (Alu) and long interspersed nucleotide element‑1 (Line‑1). This was accompanied by an increase in the expression of PDGF, and reductions of p53, PTEN and MFN2, both in mRNA and protein levels. An elevation of S‑adenosylmethionine (SAM) and a reduction of S‑adenosylhomocysteine (SAH) and the SAM/SAH ratio were also identified. In conclusion, Hcy impacted methylation the of AS‑associated genes and global methylation status that mediate the cell proliferation, which may be a character of VSMCs treated with Hcy. The data provided evidence for mechanisms of VSMCs proliferation in AS induced by Hcy and may provide a new perspective for AS induced by Hcy.

A bacterial strain, RS-LYSO-3T, was isolated from tobacco-cultivated soil, collected near Chuxiong, Yunnan province, southwestern China. RS-LYSO-3T could effectively inhibit the invasion of powdery mildew on tobacco. The colonies of RS-LYSO-3T were pale yellow, and its cells were Gram-stain-negative and rod-shaped, with 68 mol% DNA G+C content. Gene sequence analysis for its 16S rRNA gene revealed the highest similarity (97.78 %) with that of Lysobacter spongiicolaKMM 329T. Chemotaxonomic data showed that RS-LYSO-3T possesses a quinone system with Q-8, and iso-C16 : 0, summed feature 9 and iso-C15 : 0 as the predominant fatty acids, all of which support the affiliation of RS-LYSO-3T to the genus Lysobacter . The results of DNA–DNA hybridization, physiological and biochemical tests clearly proved that RS-LYSO-3T is a representative of a novel species of the genus Lysobacter , for which the name Lysobacter erysipheresistens sp. nov. is proposed. The type strain is RS-LYSO-3T (=CCIC 23922T=JCM 31042T).

In the swamps of North Myanmar lies some of the oldest stone in the world. Burmese amber (burmite) is more than 100 million years old ([ 1 ][1], [ 2 ][2]). Unlike more recent ambers from the Baltic Sea ([ 3 ][3]), Dominica ([ 4 ][4]), and India ([ 5 ][5]), burmite formed in the early Cretaceous

A Gram-stain-negative, moderately halophilic, strictly aerobic bacterium, designated YIM 95161T, was isolated from brine of a salt well in Yunnan province, China, and subjected to a taxonomic study using a polyphasic approach. Cells of strain YIM 95161T were short rods, approximately 0.9–1.4 µm long and 0.4–0.6 µm wide. Strain YIM 95161T grew at 15–40 °C (optimum, 25–30 °C), 6–29 % (w/v) NaCl (optimum, 14–19 %) and at pH 5.0–8.0 (optimum, pH 7.0). The predominant isoprenoid quinone was Q-8. The major fatty acids (>10 %) were summed feature 8 (C18 : 1ω6c and/or C18 : 1ω7c) and C14 : 0. The polar lipids consisted of diphosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, one unknown phosphoglycolipid and two unknown lipids. The DNA G+C content was 69.5 %. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain YIM 95161T was a member of the genus Salinisphaera and exhibited sequence similarities of 96.7 %, 95.6 % and 95.4 % to Salinisphaera shabanensis E1L3AT, Salinisphaera dokdonensis CL-ES53T and Salinisphaera hydrothermalis EPR70T, respectively. On the basis of phylogenetic, physiological and chemotaxonomic analysis, strain YIM 95161T represents a novel species of the genus Salinisphaera , for which the name Salinisphaera halophila sp. nov. is proposed. The type strain is YIM 95161T ( = CCTCC AB 2011132T = JCM 17431T).

A Gram-positive, moderately halotolerant, rod-shaped bacterium, designated YIM 94025(T), was isolated from a soil sample from a salt lake in Xinjiang province, north-west China. Strain YIM 94025(T) was observed to grow at 25-45 °C (optimum 37 °C), 0-22 % NaCl (optimum 2-10 %) and pH 6.0-9.0 (optimum pH 8.0). Phylogenetic analyses based on 16S rRNA gene sequences revealed that the organism belongs to the genus Tenuibacillus and exhibited sequence similarity of 98.0 % to the closest type strain, Tenuibacillus multivorans AS 1.3442(T). The predominant menaquinone was found to be MK-7; the cell-wall peptidoglycan diamino acid was meso-diaminopimelic acid; the polar lipids consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, an unidentified phospholipid and an unknown lipid; and the major fatty acids were found to contain iso-C(15:0), anteiso-C(15:0) and iso-C(16:0). The chemotaxonomic characteristics of strain YIM 94025(T) are consistent with those of the genus Tenuibacillus. The level of DNA-DNA relatedness value between YIM 94025(T) and T. multivorans AS 1.3442(T) was 36.6 ± 4.5 %. The G+C content of the strain YIM 94025(T) was determined to be 38.5 %. Based on the comparative analysis of physiological, biochemical and chemotaxonomic data, as well as DNA-DNA hybridization results, the isolate is considered to represent a novel species of the genus Tenuibacillus, for which the name Tenuibacillus halotolerans sp. nov., is proposed, with the type strain of YIM 94025(T) (=CCTCC AB 2012860(T) = KCTC 33046(T)).

Activation of FFA1 (GPR40), a member of G protein-coupling receptor family A, is mediated by medium- and long-chain fatty acids and leads to amplification of glucose-stimulated insulin secretion, suggesting a potential role for free fatty acid 1 (FFA1) as a target for type 2 diabetes. It was assumed previously that there is a single binding site for fatty acids and synthetic FFA1 agonists. However, using members of two chemical series of partial and full agonists that have been identified, radioligand binding interaction studies revealed that the full agonists do not bind to the same site as the partial agonists but exhibit positive heterotropic cooperativity. Analysis of functional data reveals positive functional cooperativity between the full agonists and partial agonists in various functional assays (in vitro and ex vivo) and also in vivo. Furthermore, the endogenous fatty acid docosahexaenoic acid (DHA) shows negative or neutral cooperativity with members of both series of agonists in binding assays but displays positive cooperativity in functional assays. Another synthetic agonist is allosteric with members of both agonist series, but apparently competitive with DHA. Therefore, there appear to be three allosterically linked binding sites on FFA1 with agonists specific for each of these sites. Activation of free fatty acid 1 receptor (FFAR1) by each of these agonists is differentially affected by mutations of two arginine residues, previously found to be important for FFAR1 binding and activation. These ligands with their high potencies and strong positive functional cooperativity with endogenous fatty acids, demonstrated in vitro and in vivo, have the potential to deliver therapeutic benefits.

This study evaluated the soil characteristics and nitrogen-fixing bacterial (NFB) communities inhabiting the rhizosphere of an invasive plant, Ageratina adenophora , in Southwest of China. In a large geographic range, heavily invaded (HI) soils generally had higher nutrient levels compared with lightly invaded (LI) and non-invaded (NI) ones. The NFB number and species diversity of HI soils were significantly greater than those of NI ones. A total of 416 NFB isolates were divided into 47 operational taxonomic units (OTUs). They were phylogenetically categorized into Proteobacteria (63.9%), Actinobacteria (32.2%), Firmicutes (1.9%) and Bacteroidetes (1.9%). At a lower taxonomic level, these isolates were grouped into 20 genera, among which, the dominant isolates were Arthrobacter , Mitsuaria , Burkholderia , Sinorhizobium , Pseudomonas and Rhizobium . The phylogenetical structures of NFB communities obtained from HI, LI and NI soils were slightly different, with their phylogenetic structure similarity indices ( θ YC ) being less than 0.8. However, the percentages of isolates represented by the shared species between any two communities were more than 80%, and in most cases, nearly 100%. There were 16 OTUs shared by these three communities (HI, NI and LI), representing 351 isolates (84.38% of total isolates). These facts indicated that numerically dominant species among HI, LI and NI soils were highly similar. In conclusion, A. adenophora invasion significantly increased the NFB number and diversity of soils, but did not change their phylogenetic structure of core community.