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The purpose of this study was to determine the neuroprotective effect and mechanism of walnut protein hydrolysates (WPH) against memory deficits induced by scopolamine in mice and further to validate the potent neuroprotective peptides identified by integrated approaches of in silico analysis and peptidomics in scopolamine-induced zebrafish. Results showed that a remarkable amelioration on behavioral performance was observed for oral administration of WPH, and disorders of cholinergic system and oxidative stress were normalized in the brains of mice also. Unfortunately, no obvious inflammatory response and anti-inflammatory effect were observed. Additionally, WPH significantly upregulated the expressions of antioxidant defense-related protein (Nrf2) and neurotrophic-related protein (BDNF and CREB). Furthermore, 20 peptides with relatively higher abundance and PeptideRanker scores were predicted by docking to AchE and Keap1. Among them, FY and SGFDAE with the highest binding affinities, -9.8 and -8.0 kcal/mol, were considered as the promising AchE and Keap1 inhibitors, respectively. They were further validated to have neuroprotective capacity in scopolamine-induced zebrafish, indicating that peptidomics and in silico prediction might be the effective approaches to screen neuroprotective peptides.

High vigor seeds have greater yield potential than those with low vigor; however, long-term storage leads to a decline in this trait. The objective of this study was to identify quantitative trait loci (QTLs) for seed vigor-related traits under artificial aging conditions using a high-density genetic linkage map of wheat (Triticum aestivum) and mine the related candidate genes. A doubled haploid population, derived from a cross between Hanxuan 10 × Lumai 14, was used as the experimental material. Six controlled-environment treatments were set up, i.e. the seeds were aged for 0, 24, 36, 48, 60, and 72 h at a high temperature (48 °C) and under high humidity (relative humidity 100%). Eight traits including seed germination percentage, germination energy, germination index, seedling length, root length, seedling weight, vigor index, and simple vigor index were measured. With the prolongation of artificial aging treatment, these traits showed a continuous downward trend and significant correlations were observed between most of them. A total of 49 additive QTLs for seed vigor-related traits were mapped onto 12 chromosomes (1B, 2D, 3A, 3B, 3D, 4A, 4D, 5A, 5B, 5D, 6D, and 7A); and each one accounted for 6.01–17.18% of the phenotypic variations. Twenty-five pairs of epistatic QTLs were detected on all chromosomes, except for 5D, 6A, and 7D, and each epistasis accounted for 7.35–26.06% of the phenotypic variations. Three additive QTL hot spots were found on chromosomes 5A, 5B, and 5D, respectively. 13 QTLs, QGEe5B, QGIe5B, QSLc5B, QSLd5B, QSLf5B, QRLd5B, QRLe5B, QRLf5B, QVId5B, QVIe5B, QVIf5B, QSVId5B, and QSVIe5B, were located in the marker interval AX-94643729 ~ AX-110529646 on 5B and the physical interval 707,412,449–710,959,479 bp. Genes including TRAESCS5B01G564900, TRAESCS5B01G564200, TRAESCS5B01G562600, TraesCS5B02G562700, TRAESCS5B01G561300, TRAESCS5B01G561400, and TRAESCS5B01G562100, located in this marker interval, were found to be involved in regulating the processes of carbohydrate and lipid metabolism, transcription, and cell division during the germination of aging seeds, thus they were viewed as candidate genes for seed viability-related traits. These findings provide the basis for the seed-based cloning and functional identification of related candidate genes for seed vigor.

Background Human demand for wheat will continue to increase together with the continuous global population growth. Agronomic traits in wheat are susceptible to environmental conditions. Therefore, in breeding practice, priority is given to QTLs of agronomic traits that can be stably detected across multiple environments and over many years. Results In this study, QTL analysis was conducted for eight agronomic traits using an introgression line population across eight environments (drought stressed and well-watered) for 5 years. In total, 44 additive QTLs for the above agronomic traits were detected on 15 chromosomes. Among these, qPH-6A, qHD-1A, qSL-2A, qHD-2D and qSL-6A were detected across seven, six, five, five and four environments, respectively. The means in the phenotypic variation explained by these five QTLs were 12.26, 9.51, 7.77, 7.23, and 8.49%, respectively. Conclusions We identified five stable QTLs, which includes qPH-6A, qHD-1A, qSL-2A, qHD-2D and qSL-6A. They play a critical role in wheat agronomic traits. One of the dwarf genes Rht14, Rht16, Rht18 and Rht25 on chromosome 6A might be the candidate gene for qPH-6A. The qHD-1A and qHD-2D were novel stable QTLs for heading date and they differed from known vernalization genes, photoperiod genes and earliness per se genes.

Nitrogen plays a crucial role in wheat growth and development. Here, we analyzed the tolerance of wheat strains XM26 and LM23 to low-nitrogen stress using a chlorate sensitivity experiment. Subsequently, we performed transcriptome analyses of both varieties exposed to low-nitrogen (LN) and normal (CK) treatments. Compared with those under CK treatment, 3534 differentially expressed genes (DEGs) were detected in XM26 in roots and shoots under LN treatment (p &lt, 0.05, and |log2FC| &gt, 1). A total of 3584 DEGs were detected in LM23. A total of 3306 DEGs, including 863 DEGs in roots and 2443 DEGs in shoots, were specifically expressed in XM26 or showed huge differences between XM26 and LM23 (log2FC ratio &gt, 3). These were selected for gene ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses. The calcium-mediated plant–pathogen interaction, MAPK signaling, and phosphatidylinositol signaling pathways were enriched in XM26 but not in LM23. We also verified the expression of important genes involved in these pathways in the two varieties using qRT-PCR. A total of 156 transcription factors were identified among the DEGs, and their expression patterns were different between the two varieties. Our findings suggest that calcium-related pathways play different roles in the two varieties, eliciting different tolerances to low-nitrogen stress.

Trehalose-6-phosphate phosphatase (TPP) genes take part in trehalose metabolism and also in stress tolerance, which has been well documented in many species but poorly understood in wheat. The present research has identified a family of 31 TPP genes in Triticum aestivum L. through homology searches and classified them into five clades by phylogenetic tree analysis, providing evidence of an evolutionary status with Hordeum vulgare, Brachypodium distachyon and Oryza sativa. The exon-intron distribution revealed a discrete evolutionary history and projected possible gene duplication occurrences. Furthermore, different computational approaches were used to analyze the physical and chemical properties, conserved domains and motifs, subcellular and chromosomal localization, and three-dimensional (3-D) protein structures. Cis-regulatory elements (CREs) analysis predicted that TaTPP promoters consist of CREs related to plant growth and development, hormones, and stress. Transcriptional analysis revealed that the transcription levels of TaTPPs were variable in different developmental stages and organs. In addition, qRT-PCR analysis showed that different TaTPPs were induced under salt and drought stresses and during leaf senescence. Therefore, the findings of the present study give fundamental genomic information and possible biological functions of the TaTPP gene family in wheat and will provide the path for a better understanding of TaTPPs involvement in wheat developmental processes, stress tolerance, and leaf senescence.

The mechanisms for physiological senescence in bamboo shoots after harvest remain unclear. This study investigated physiological changes in Fargesia yunnanensis shoots during storage at different temperatures. The relationship between morphological and physiological changes in bamboo shoots during storage was also analyzed. The results show that cold storage can reduce weight loss, browning, respiration rates, and sugar degradation in bamboo shoots; decrease related enzymatic activities; and inhibit the increase in lignin and cellulose content. The quality of bamboo shoots declines more during the first 3d after harvesting than it does during subsequent periods. The increase in the degree of lignification and fibrosis is the main reason for senescence and for the decline in quality of bamboo shoots after harvest. The bamboo shoots under RT conditions began browning during the third 3d of storage, with a browning rate of 688gkg−1 even in the upper parts; the increase in shoot browning degrees significantly decreased the quality. Low temperatures had better inhibitory effects on browning than they did on lignification and fibrosis. Nonstructural carbohydrates in bamboo shoots are degraded and flow into sheath and shoot respiration, phenols, and shoot fibrosis and lignification at room temperature, but only flow into sheath respiration, shoot fibrosis, and lignification at cold temperature. Soluble protein and free amino acids are primarily distributed into shoot and sheath respiration and into phenols at room temperature, but that process is well inhibited at cold temperature. Bamboo shoots, once removed from cold storage, should be consumed rapidly because enzyme activity recovers quickly. This research provides new theoretical information on the preservation of bamboo shoots.

Mesenchymal stem cell (MSC) therapy is considered one of the most promising treatments in the context of the coronavirus disease 2019 (COVID-19) pandemic. However, the safety and effectiveness of MSCs in the treatment of COVID-19-associated pneumonia patients need to be systematically reviewed and analyzed. Two independent researchers searched for relevant studies published between October 2019 and April 2021 in the PubMed, Embase, Cochrane Library, WAN FANG, and CNKI databases. All relevant randomized controlled trials, clinically controlled studies, retrospective studies, case reports, letters (with valid data), and case series were included in this meta-analysis. A fixed-effects model and 95% confidence interval (CI) were used to analyze the results. A total of 22 studies involving 371 patients were included in the present study. Allogeneic MSCs from umbilical cord, adipose tissue, menstrual blood, placental tissue, Wharton's jelly, or unreported sources were administered in 247 participants. Combined results revealed that MSC therapy significantly reduced the incidence of adverse events [AEs; odds ratio (OR) = 0.43, 95% CI = 0.22-0.84

Silicon is an essential element for the growth and development of bamboo plants that significantly affects the biochemical and physical properties for industrialized bamboo utilization. Silicon is biomineralized as diverse forms of phytoliths in the bamboo organs. We investigated the silicon variation and phytolith morphology in different organs of Dendrocalamus brandisii (Munro) Kurz. The silicon content in D. brandisii (Munro) Kurz varies and increases in the order of leaf > sheath > branch > culm > shoot > root. The phytolith morphotypes and presence frequency in different D. brandisii organs varied closely interrelated with the cell structure and organ functions. Amorphous silicon deposition and few varieties and numbers of phytolith were found in D. brandisii root and shoots. Rich distribution of the saddle and dumb-bell phytoliths was found in all organs except the root and mature culm and branch. More elongate and rectangular phytoliths were found in mature culm and branch. Round and elliptical phytoliths might be firstly formed followed by dumb-bell and saddle phytoliths. Phytolith sizes increased with increasing culm, branch and leaf age, but did not demonstrate a specific trend with culm height. The result could contribute to enriching phytolith morphology in Gramineae and provide theoretical basis and references for plant taxonomy and large-scale bamboo industrial processing and utilization.

Background Premature senescence of flag leaf severely affects wheat yield and quality. Chlorophyll (Chl) degradation is the most obvious symptom during leaf senescence and catalyzed by a series of enzymes. Pheophytin pheophorbide hydrolase (Pheophytinase, PPH) gene encodes a Chl degradation hydrolase. Results In this study, the coding, genomic and promoter sequences of wheat TaPPH-A gene were cloned. The corresponding lengths were 1467 bp, 4479 bp and 3666 bp, respectively. Sequence structure analysis showed that TaPPH-A contained five exons and four introns. After the multiple sequences alignment of TaPPH-A genome from 36 accessions in a wheat diversity panel, four SNPs and one 2-bp InDel were observed, which formed two haplotypes, TaPPH-7A-1 and TaPPH-7A-2. Based on the SNP at 1299 bp (A/G), a molecular marker TaPPH-7A-dCAPS was developed to distinguish allelic variation (A/G). Using the molecular markers, 13 SSR, and 116 SNP markers, a linkage map of chromosome 7A were integrated. TaPPH-A was mapped on the chromosome region flanked by Xwmc9 (0.94 cM) and AX-95634545 (1.04 cM) on 7A in a DH population. Association analysis between TaPPH-7A allelic variation and agronomic traits found that TaPPH-7A was associated with TGW in 11 of 12 environments and Chl content at grain-filling stage under drought stress using Population 1 consisted of 323 accessions. The accessions possessed TaPPH-7A-1 (A) had higher TGW and Chl content than those possessed TaPPH-7A-2 (G), thus TaPPH-7A-1 (A) was a favorable allelic variation. By analyzing the frequency of favorable allelic variation TaPPH-7A-1 (A) in Population 2 with 157 landraces and Population 3 with 348 modern cultivars, we found it increased from pre-1950 (0) to 1960s (54.5%), then maintained a relatively stable level about 56% from 1960s to 1990s. Conclusion These results suggested the favorable allelic variation TaPPH-7A-1 (A) should be valuable in enhancing grain yield by improving the source (chlorophyll content) and sink (the developing grain) simultaneously. Furthermore, the newly developed molecular marker TaPPH-7A-dCAPS could be integrated into a breeding kit of screening high TGW wheat for marker-assisted selection. Electronic supplementary material The online version of this article (10.1186/s12870-019-1901-0) contains supplementary material, which is available to authorized users.

Senescence in a wheat (Triticum aestivum L.) leaf is a programmed degeneration process leading to death. During this process, green leaf area duration (GLAD) and green leaf number of main stem (GLNMS) are gradually reduced. In this study, the two traits of Hanxuan10/Lumai14 DH population at different development stages after anthesis were evaluated under rainfed and irrigated conditions, and QTLs were detected. GLAD and GLNMS of two parents and DH population under rainfed condition were less than those under irrigated condition, and close correlations (P < 0 05) were found between GLAD and GLNMS after 25 DAA under both water conditions. GLAD and GLNMS were co-controlled by major and minor genes. QTLs for GLAD were stably expressed at different development stages after anthesis under both water conditions, such as QGlad22-1B-1, QGlad25-1B-1, QGlad28-1B-2 detected under irrigated condition and QGlad25-1B-3, QGlad28-1B-4 mapped under rainfed condition were located at a 20.7 cM marker interval of Xgwm273-EST122 on 1B chromosome. But QTLs for GLNMS were inducibly and specifically expressed at specific developmental stages after anthesis under both water conditions. The findings provide dynamic genetic information related to wheat senescence.

Bamboo is one of the fastest growing plants in the world, but their shoot buds develop very slowly. Information about the sugar storage and metabolism during the shoot growth is lacking. In the present study, we determined the activity of sucrose and starch metabolizing enzymes during the developmental period of Fargesia yunnanensis from shoot buds to the young culms that have achieved their full height. The soluble sugars and starch contents were also determined and analyzed in shoot buds and shoots at different developmental stages. The results showed that there were higher sucrose contents in shoot buds than shoots, which coincides with the sweeter taste of shoot buds. As the shoot buds sprouted out of the ground, the starch and sucrose were depleted sharply. Coupled with this, the activity of soluble acid invertase (SAI), cell wall-bound invertase (CWI), sucrose synthase at cleavage direction (SUSYC) and starch phosphorylase (STP) increased significantly in the rapidly elongating internodes. These enzymes dominated the rapid elongation of internodes. The activities of SAI, CWI, SUSYC and STP and adenosine diphosphate-glucose pyrophosphorylase were higher as compared to other enzymes in the shoot buds, but were far lower than those in the developing shoots. The slow growth of shoot buds was correlated with the low activity of these enzymes. These results complement our understanding of the physiological differences between shoot buds and elongating shoots and ascertain the physiological mechanism for the rapid growth of bamboo shoots.

Purpose Angiogenesis actively contributes to tumor growth and metastasis. MACC1 was reported to be associated with tumor progression. In the present study, we aimed to investigate the expression and role of MACC1 in cholangiocarcinoma (CCA) and its correlation with angiogenesis. Patients and methods We investigated the expression and correlation of MACC1 and VEGFA in The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) datasets and in 7 paired frozen CCA and matched paracarcinoma tissues. Immunohistochemistry (IHC) was used to examine MACC1 and VEGFA expression as well as microvessel density (MVD) in 122 paraffin-embedded CCA samples. Western blotting, real-time qPCR and ELISA were performed to investigate the effect of MACC1 knockdown on VEGFA expression and secretion in CCA cells. Subsequently, we collected conditioned medium from cells with MACC1 knockdown and used it in angiogenesis assays. Results The expression levels of both MACC1 and VEGFA were significantly upregulated in the TCGA and GEO datasets and in the 7 paired frozen CCA tissues compared to the matched paracarcinoma tissues, and MACC1 was significantly correlated with VEGFA. IHC showed that high expression of MACC1 and VEGFA was significantly correlated with lymph node metastasis (P

Sleep deprivation negatively influences memory formation and consolidation, which leads to memory impairment associated with oxidative stress and neurotrophic pathways. In this study, a sleep-deprived rat model was used to evaluate the protective effects of round scad hydrolysates (RSH, 333 and 666 mg per kg bw) on memory impairment and the underlying mechanisms. The result of the Morris water maze test revealed that RSH significantly reversed the cognition deficits induced by sleep deprivation. Moreover, RSH supplementation alleviated oxidative stress by increasing the activities of glutathione peroxidase and superoxide dismutase and the ratio of glutathione/oxidized glutathione in the brain. Furthermore, RSH significantly up-regulated the expression of antioxidant defense-related proteins, including nuclear factor E2-related factor 2 (Nrf2) and its downstream heme oxygenase-1 (HO-1), as well as the phosphorylation of Akt in rats. Moreover, RSH improved the expression of brain-derived neurotrophic factor (BDNF), the phosphorylation of cAMP response element-binding (CREB) and tropomyosin-related kinase B (TrkB) in vivo, indicating that RSH can also promote the expression of proteins related to the neurotrophic pathway. Additionally, UPLC-qTOF-MS/MS was further used to identify the peptides in RSH. The results indicated that RSH mainly consists of low molecular weight peptides with hydrophobic, aromatic and positively charged amino acids in sequence. Taken together, these findings demonstrate that RSH exerts memory-improving actions by regulating the antioxidant and neurotrophic pathways, and RSH can be a potential functional ingredient for the prevention and protection of cognitive deficits.

The peptide derived from Anchovy hydrolysates, Pro-Ala-Tyr-Cys-Ser (PAYCS), was reported to display a neuroprotective effect in vitro in our previous study. The in vivo memory improving effects of PAYCS were investigated in this study. Prior to the scopolamine-induced amnesia mice trial, the stability of PAYCS during digestion was detected and the digestive products were identified. The results showed that PAYCS was susceptible to proteolytic degradation after incubation with pepsin and pancreatin and Pro-Ala-Tyr (PAY) was released and survived during the simulated GI digestion. The results of scopolamine-induced amnesia model trials showed that PAYCS and PAY treatment exhibited cognitive improvement effects in the behavioral tests and different pathways were determined. The results indicated that only PAYCS facilitated cholinergic activity by up-regulating the amount of acetylcholine (Ach) and acetylcholine receptor (AChR). Additionally, both PAYCS and PAY enhanced the superoxide dismutase (SOD) activity. Furthermore, PAYCS was found to be beneficial for the expression of the nuclear factor (erythroid-derived2)-like 2 protein (Nrf2), brain-derived neurotrophic factor (BDNF) and cAMP response element binding protein (CREB). This indicated that PAYCS could regulate the oxidative stress by activating the Nrf2/antioxidant response elements (Nrf2/ARE) pathway. In our study, we demonstrated that the memory improving effects conferred by PAYCS on amnesia mice were linked to the attenuation of the cholinergic system and the activation of Nrf2/ARE and BDNF/CREB signaling.

BackgroundThe specification and differentiation of neocortical projection neurons is a complex process under precise molecular regulation; however, little is known about the similarities and differences in cerebral cortex development between human and mouse at single-cell resolution.ResultsHere, using single-cell RNA-seq (scRNA-seq) data we explore the divergence and conservation of human and mouse cerebral cortex development using 18,446 and 7,610 neocortical cells. Systematic cross-species comparison reveals that the overall transcriptome profile in human cerebral cortex is similar to that in mouse such as cell types and their markers genes. By single-cell trajectories analysis we find human and mouse excitatory neurons have different developmental trajectories of neocortical projection neurons, ligand-receptor interactions and gene expression patterns. Further analysis reveals a refinement of neuron differentiation that occurred in human but not in mouse, suggesting that excitatory neurons in human undergo refined transcriptional states in later development stage. By contrast, for glial cells and inhibitory neurons we detected conserved developmental trajectories in human and mouse.ConclusionsTaken together, our study integrates scRNA-seq data of cerebral cortex development in human and mouse, and uncovers distinct developing models in neocortical projection neurons. The earlier activation of cognition -related genes in human may explain the differences in behavior, learning or memory abilities between the two species.

The flag leaf is the main organ of photosynthesis during grain-filling period of wheat, and flag leaf-related traits affect plant morphology and yield potential. In this study, two BC3F6 introgression line (IL) populations derived from the common recipient parent Lumai 14 with Jing 411 and Shaanhan 8675, respectively, were used to map quantitative trait loci (QTL) for flag leaf length (FLL), flag leaf width (FLW), flag leaf area (FLA) and chlorophyll content (CC) at flowering stage and 15 and 20 days after anthesis (DAA) in 2016–2017 (E1) and 2017–2018 (E2) two environments. A total of 14 and 15 QTLs for flag leaf-related traits were detected in Lumai 14 / Jing 411 and Lumai 14 / Shaanhan 8675 populations, respectively. Among them, Both QFLW-6A and QFLA-6A were detected in Lumai 14 / Jing 411 population under E2 and in Lumai 14 / Shaanhan 8675 population under E1 and E2 environments, respectively. QCCS2-3A from Lumai 14 / Jing 411 population and QCCS3-1A, QFLL-4A and QFLL-6A from Lumai 14 / Shaanhan 8675 population were repeatedly identified under two tested environments. Moreover, eight QTL clusters controlling flag leaf-related traits were identified, which provided a genetic basis for significant correlations in phenotype among these traits. On the other hand, positive alleles of QFLW-6A for FLW detected in two populations were derived from their donors. Eighteen lines and 44 lines carried this QTL were found in Lumai 14 / Jing 411 and Lumai 14 / Shaanhan 8675 populations, respectively. The means of FLW in these lines were wider than that of the recipient parent, Lumai 14, in two environments, suggesting that QFLW-6A played an important role for increasing FLW. The IL 124 in Lumai 14 / Jing 411 population and the IL 59 and IL 127 in Lumai 14 / Shaanhan 8675 population had five, five and four donor chromosomal segments which carried no other QTL controlling FLW than QFLW-6A, respectively. And the FLWs of these lines were significantly greater than that of Lumai 14 under two environments. So these lines and their donor parent can be regarded as potential near-isogenic lines. Further, a synteny analysis found QFLW-6A was near the 574,283,851–574,283,613 bp fragment on chromosome 6A and 10 genes were in the range of 500 kb upstream and downstream of the fragment. These results provide the basis for identification of candidate gene and map-based cloning and functional verification of the QTL.

Among multiple ways to ameliorate Alzheimer's disease, antioxidant and acetylcholinesterase (AChE) inhibition are considered as two primary approaches. Previous studies have indicated that anchovy protein hydrolysate (APH) exhibited strong antioxidant and AChE inhibitory capacities, and Maillard reaction could significantly (p

Urban wetlands play an important role in improving urban environment and microclimate, while they are suffering degradation and destruction during urbanization process. Arbuscular mycorrhizae may enhance plant tolerance to environmental stresses and increase vegetation restoration in wetlands. However, presently little is known how about arbuscular mycorrhizal (AM) symbiosis in urban wetland plants and the factors affecting AM formation. Here, a survey for AM status in urban wetland plants was done in Beijing area, and the factors affecting AM formation were also discussed, such as water and sediment characteristics. Results showed that 87.5% of plants (49 of 56 species) were colonized by AM fungi. Mycorrhizal colonization rates (MCRs) ranged from 2% to 72%, while most of them were low level (

Culm sheath is an important morphological characteristic of bamboos; however, its physiological functions are still unknown. The main goal of this study was to examine the physiological functions of culm sheaths and their effects on the height growth of Fargesia yunnanensis Hsueh et Yi shoots. The elongating internodes were analyzed for determining the soluble sugar and starch contents, as well as the water potential of different sections. The transpiration rates were also determined and compared between culm sheaths and foliage leaf blades. The effects of culm sheaths on the shoot height growth were also analyzed. The results showed that the outer zone had higher starch but lower soluble sugar contents than the inner zone in the developing shoots. The transpiration rate of culm sheaths was far higher than that of leaves. The water potential of the outer zone was lower than that of the inner zone in bamboo culms. The growth of bamboo culms decreased without culm sheaths. When one strip of culm sheaths was peeled off longitudinally from young culms, the bare side of the culms curved rapidly after one night, and the water potential gradient between the inner and outer zones significantly decreased, further demonstrating that sheaths have significant effects on bamboo culm height growth.

Wear debris-induced osteolysis is one of the major reasons for subsequent aseptic loosening after cementless hip arthroplasty. Increasing evidence suggests that receptor activator of nuclear factor kappa-B (NF-κB) ligand-mediated osteoclastogenesis and osteolysis are responsible for wear debris-induced aseptic loosening. In the present study, we explored the effect of amentoflavone (AMF) on inhibiting osteoclast generation and wear debris-induced osteolysis in vitro and in vivo. Twenty-four male C57BL/J6 mice were randomly divided into four groups: a sham group and groups with titanium wear debris treatment followed by intraperitoneal injection of various concentrations of AMF (0, 20, and 40 mg/kg/day). The micro computed tomography scanning and histological analysis were performed. Bone marrow-derived macrophages were cultured to investigate the effect of AMF on osteoclast generation and function. The results showed that AMF suppressed osteoclastogenesis, F-actin ring formation, and bone absorption without cytotoxicity. AMF prevented titanium wear debris-induced osteolysis in mice. AMF suppressed the relative proteins of NF-κB and mitogen-activated protein kinase (MAPKs) signaling pathways. Thus, the present study suggests that AMF derived from plants could inhibit osteoclastogenesis and titanium wear debris-induced osteolysis via suppressing NF-κB and MAPKs signaling pathways.

Cross-linked enzyme aggregates (CLEAs) have emerged as an interesting biocatalyst design for enzyme immobilization. However, the commercialization of CLEAs is often hampered by their shortcomings, such as poor-controlled particle size, low activity and sticky characteristic. In order to overcome these drawbacks, five nanoparticles (NPs) (nano-TiO2, nano-MgO, nano-Ni, nano-Cu and nano-Fe3O4) were used to improve CLEAs activity and structure in this study. Results showed that moderate dosage of nano-TiO2 addition increased CLEAs activity, and the most increment was 15.2% relative to CLEAs without NPs. This was possibly due to more channels and smaller size particle of CLEAs after nano-TiO2 addition. Moreover, nano-TiO2 addition not only decreased fluorescence intensity but also caused the red shift in tryptophan (Trp) and tyrosine (Typ) residues. Nano-TiO2 addition decreased Km and increased Vmax of CLEAs. These changes were benefit to the activity and structure of CLEAs. However, addition of other four NPs (nano-MgO, nano-Ni, nano-Cu and nano-Fe3O4) did not increase CLEAs activity and even decrease CLEAs activity due to less amorphous cavities and larger or discrete particle size than CLEAs without NPs. In addition, FT-IR results showed that NPs addition increases the content of regular structure of CLEAs, and causes a partial transformation of β-turn into β-sheet. This study showed that it was potential to improve CLEAs performance by nano-TiO2 addition.

Tacrolimus (FK506) has been demonstrated to reduce epidural fibrosis. However, the detailed mechanism of action has not been elucidated. Aberrant miR-429 is involved in many diseases. The aim of this study was to describe the exact mechanism of FK506 induced apoptosis in fibroblasts and the prevention of epidural fibrosis. FK506 induced fibroblast apoptosis was evaluated using CCK-8 assays, flow cytometry, and western blotting. The expression of miR-429 in fibroblasts treated with FK506 was determined by RT-qPCR. Additionally, luciferase activity assays were used to determine the target relationship between miR-429 and RhoE. Flow cytometry and western blot analysis were used to determine the effects of FK506 and miR-429 on fibroblast apoptosis. The effects of FK506 and RhoE on fibroblast apoptosis were determined by CCK-8 assay, flow cytometry, and western blotting. We also evaluate the effects of FK506 and miR-429 on epidural fibrosis in rats by using histological analysis and TUNEL-staining. The results revealed FK506 induces fibroblast apoptosis and significantly downregulates miR-429 expression in fibroblasts. Additionally, miR-429 downregulation caused the apoptosis of fibroblasts. The luciferase activity assay confirmed that RhoE is a direct target of miR-429 and RhoE promotes fibroblast apoptosis. The rat model demonstrated miR-429 inhibition promotes fibroblast apoptosis and epidural fibrosis, which is consistent with the results of FK506 treatment. Our study demonstrates that FK506 induces fibroblast apoptosis and reduces epidural fibrosis by regulating miR-429 expression and its target of RhoE.

Background Circular RNAs (circRNAs) have recently been shown to play important roles in different tumors. However, their detailed roles and regulatory mechanisms in pancreatic ductal adenocarcinoma (PDAC) are not well understood. This study aimed to identify enriched circRNAs and detect their functions and mechanisms in PDAC cells and tissues. Methods circRNA-ASH2L (circ-ASH2L) was identified by circRNA microarray studies based on previous studies, and further detected in PDAC cells and samples by qRT-PCR. The functions of circ-ASH2L were identified by transwell, EdU, cell cycle or Tube formation assays. The regulatory mechanisms of circ-ASH2L were explored by WB, RIP, FISH, dual-luciferase assays, RNA pulldown or other assays. Results We identified a circRNA (circ-ASH2L) based on our previous studies, detected its expression in different malignant cells and found that circ-ASH2L was highly expressed in pancreatic cells or tumor tissues and correlated with tumor malignancy. Further studies revealed that circ-ASH2L promoted tumor invasion, proliferation and angiogenesis by regulating miR-34a, thus regulate Notch 1 expression. Circ-ASH2L served as a miRNA sponge for miR-34a and promoted tumor progression in vivo. Finally, we analyzed circ-ASH2L expression in clinical tissues and found that high circ-ASH2L expression was correlated with lymphatic invasion and TNM stage and was an independent risk factor for pancreatic patient survival. Conclusions circ-ASH2L play an important role in tumor invasion, and high circ-ASH2L may be a useful marker of PDAC diagnosis or progression.

Epilepsy is a common neurological disorder that affects mammalian species including human beings and dogs. In order to discover novel drugs for the treatment of canine epilepsy, multiomics data were analyzed to identify epilepsy associated genes. In this research, the original ranking of associated genes was obtained by two high-throughput bioinformatics experiments: Genome Wide Association Study (GWAS) and microarray analysis. The association ranking of genes was enhanced by a re-ranking system, HPO-Shuffle, which integrated information from GWAS, microarray analysis and Human Phenotype Ontology database for further downstream analysis. After applying HPO-Shuffle, the association ranking of epilepsy genes were improved. Afterward, a weighted gene coexpression network analysis (WGCNA) led to a set of gene modules, which hinted a clear relevance between the high ranked genes and the target disease. Finally, WGCNA and connectivity map (CMap) analysis were performed on the integrated dataset to discover a potential drug list, in which a well-known anticonvulsant phensuximide was included.

Cholangiocarcinoma (CCA) is the second most common hepatobiliary cancer after hepatocellular carcinoma. Antiangiogenic therapy has been administered to patients with CCA, but the benefits of this therapy remain unsatisfactory. Improved understanding of the molecular mechanisms underlying angiogenesis in CCA is required. In the present study, the expression of GATA‑binding protein 6 (GATA6), lysyl oxidase‑like 2 (LOXL2) and vascular endothelial growth factor A (VEGFA), in addition to the microvessel density (MVD), were evaluated by performing immunohistochemical staining of human CCA microarrays. The expression of GATA6/LOXL2 was associated with poor overall survival (P=0.01) and disease‑free survival (P=0.02), and was positively associated with VEGFA expression (P=0.02) and MVD (P=0.04). In vitro, western blotting, reverse transcription‑quantitative PCR analysis and ELISAs revealed that altered GATA6 and LOXL2 expression regulated the expression levels of secreted VEGFA. Co‑immunoprecipitation demonstrated a physical interaction between GATA6 and LOXL2 in CCA cell lines, and the scavenger receptor cysteine‑rich domain of LOXL2 interacted with GATA6, which regulated VEGFA mRNA expression and protein secretion, and promoted tube formation. In vivo analyses further revealed that GATA6/LOXL2 promoted VEGFA expression, angiogenesis and tumor growth. The GATA6/LOXL2 complex represents a novel candidate prognostic marker for stratifying patients with CCA. Drugs targeting this complex may possess great therapeutic value in the treatment of CCA.

The three genetic populations used in this study included a doubled-haploid (DH) population derived from ‘Hanxuan 10’ × ‘Lumai 14’, a BC3F6 population from ‘Lumai 14’ × ‘Jing 411’ [introgression line (IL) population 1], and a BC3F6 population from ‘Lumai 14’ × ‘Shaanhan 8675’ (IL population 2). The genetic characters underpinning kernel morphological traits, such as kernel length, kernel width, kernel thickness, kernel length/width ratio, kernel length/thickness ratio, and kernel width/thickness ratio were analyzed. Quantitative trait loci (QTL) for all the above traits were mapped in the three populations across six, three, and three environments, respectively. The genetic effects of qKW-6A, which was detected in all three populations, were analyzed. Forty six additive QTLs for kernel morphological traits were detected in the DH population, and 20 additive QTLs were detected in each of the IL populations. A kernel-width QTL, qKW-6A, was located within the same interval in all three populations. qKT-7A-3, qLTR-4A, and qWTR-7A-1 mapped in the DH population were located in the same marker intervals as qKT-7A-1, qLTR-4A, and qWTR-7A-1, respectively, in IL population 2. qLWR-5A-2, qWTR-5A-2, and qWTR-5A-1 from the DH population were detected in five, four, and three environments, and explained 14.72, 25.11, and 25.91%, respectively, of the phenotypic variation. qLTR-7A from IL population 1 and qLWR-5B from IL population 2, detected in all three environments, explained 6.10 and 10.66% of the phenotypic variation, respectively. On the other hand, negative alleles of qKW-6A for kernel width detected in all three populations were derived from ‘Lumai 14’. Donor segments including this QTL were introgressed into 18 lines of IL population 1 and 44 lines of IL population 2. The mean kernel width in these lines was greater than the recurrent parent ‘Lumai 14’ under drought-stress conditions in 2 years, indicating that qKW-6A played an important role in determination of kernel width. Line 157 from IL population 2 contained only five chromosomal segments from the donor parent, and these donor segments harbored no QTL for kernel width other than qKW-6A. However, kernel width in this line was significantly greater than that of ‘Lumai 14’ in all three environments. Thus, Line 157 can be regarded as a near-isogenic line for fine mapping and map-based cloning of qKW-6A.

Efficient single-cell assignment is essential for single-cell sequencing data analysis. With the explosive growth of single-cell sequencing data, multiple single-cell sequencing data sources are available for the same kind of tissue, which can be integrated to further improve single-cell assignment; however, an efficient integration strategy is still lacking due to the great challenges of data heterogeneity existing in multiple references. To this end, we present mtSC, a flexible single-cell assignment framework that integrates multiple references based on multitask deep metric learning designed specifically for cell type identification within tissues with multiple single-cell sequencing data as references. We evaluated mtSC on a comprehensive set of publicly available benchmark datasets and demonstrated its state-of-the-art effectiveness for integrative single-cell assignment with multiple references.

The Bambusoideae subfamily includes the fastest-growing plants worldwide, as a consequence of fast internode elongation. However, few studies have evaluated the temporal and spatial distribution of endogenous hormones during internode elongation. In this paper, endogenous indole-3-acetic acid (IAA) and abscisic acid (ABA) were detected in different developmental internodes during shoot elongation by immunolocalization. Immunohistochemistry showed that IAA was mainly present in the shoot apex, leaf sheath primordia, parenchymal cells, and vascular tissues. During internode elongation and maturation, the IAA signals decreased significantly and then increased slightly, with the weakest signals observed in the rapidly elongating internode. Based on immunogold localization, most IAA signals were detected in the cytoplasm and nuclei of both parenchymal and fiber cells, and few signals were detected in cell walls in the unelongated and elongating internodes. After the completion of internode elongation, additional IAA signals were detected in the secondary walls of both parenchymal and fiber cells. Immunohistochemical localization of ABA showed that ABA signals decreased with internode elongation and maturation, with the weakest signal observed in the internodes of 3-month-old shoots. In addition, few ABA signals were detected in the shoot apex. The strongest IAA and ABA signals in unelongated internodes suggested that both hormones participated in the mediation of internode differentiation but not in the rapid elongation. Moreover, IAA was involved in secondary cell wall deposition.

Stomatal density and size affect plant water use efficiency, photosynthsis rate and yield. The objective of this study was to gain insights into the variation and genetic basis of stomatal density and size during grain filling under drought stress (DS) and well-watered (WW) conditions. The doubled haploid population derived from a cross of wheat cultivars Hanxuan 10 (H10), a female parent, and Lumai 14 (L14), a male parent, was used for phenotyping at the heading, flowering, and mid- and late grain filling stages along with established amplified fragment length polymorphism (AFLP) and simple sequence repeat (SSR) markers. The stomatal density of doubled haploid (DH) lines was gradually increased, while the stomatal lengths and widths were gradually decreased during grain filling stage. Twenty additive QTLs and 19 pairs of epistatic QTLs for the 3 traits were identified under DS. The other 20 QTLs and 25 pairs epistatic QTLs were obtained under WW. Most QTLs made more than 10% contributions to the total phenotypic variations at one growth stage under DS or WW. Furthermore, QTLs for stomatal density near Xwmc74 and Xgwm291 located on chromosome 5A were tightly linked to previously reported QTLs regulating total number of spikelets per spike, number of sterile spikelets per spike and proportion of fertile spikelets per spike. Qsw-2D-1 was detected across stages, and was in the same marker region as a major QTL for plant height, QPH. cgb-2D.1 . These indicate that these QTLs on chromosomes 5A and 2D are involved in regulating these agronomic traits and are valuable for molecular breeding.

As of today, the functions of fusoid cell, and the transport and loading pathways of photoassimilate in bamboo leaves are still not clear. In this paper, the leaves of Fargesia yunnanensis from a greenhouse and the wild were respectively used as samples to analyze the anatomical characteristics of fusoid cells and vascular bundles. The results showed that the bamboo leaves from greenhouse got shorter and thinner with fewer layers of palisade parenchyma cells than those from the wild. The volumes of fusoid cells were also increased. Fusoid cells originated from a huge parenchyma cell as testified by the observed nuclei. Several fusoid cells usually formed one cell complex close to the midrib. Crystals were detected in fusoid cells but no pits or plasmodesmata on their walls, suggesting that fusoid cells had the function of regulating water. The presence of fusoid cells determined the major difference between a leaf blade and sheath. There were prominent chloroplasts with simple stroma lamellae in the parenchymatous bundle sheath cells and starch grains were also observed in these chloroplast. Photoassimilates could be transported across vascular bundle sheath via symplasmic pathways for an abundant of plasmodesmata in sheath cell walls, and transported into phloem tube by apoplastic pathway as there were no pits in the walls of companion cells and phloem tubes.

BACKGROUND The usefulness of Des-g-carboxyprothrombin (DCP) has been indicated in areas where hepatitis C virus is prevalent. DCP has yet to be used in China. The aim of this study was to evaluate the usefulness of DCP in Chinese patients with hepatocellular carcinoma (HCC) predominantly caused by hepatitis B. MATERIAL AND METHODS 329 subjects with HCC and 371 subjects without HCC that all underwent surgery were consecutively enrolled. Serum AFP and plasma DCP levels in all subjects and 153 healthy volunteers were measured and analyzed. RESULTS Of 329 subjects with HCC, 258 (78.4%) were HBsAg positive. The median level of plasma DCP was 853.72 mAU/mL in subjects with HCC, 26.43 mAU/mL in subjects without HCC, and 29.91 m AU/mL in healthy volunteers. A cut-off DCP value of 87 mAU/mL yielded the optimal sensitivity of 74.80% and a specificity of 83.33% for differentiating subjects with HCC from subjects without HCC. The combination of AFP of 21.33 ng/mL and DCP of 87 mAU/mL had a sensitivity of 82.60% for tumors no larger than 2 cm, as well as a sensitivity of 90% for tumors larger than 5 cm. CONCLUSIONS The combination of DCP and AFP yielded great improvement in sensitivity in differentiating subjects with HCC from subjects without HCC. These two markers may be incorporated in the protocol for surveillance and diagnosis of HCC in the high-risk Chinese population.

B7-H4, as a member of the costimulatory B7 family, serves a critical role in the negative regulation of T cell-mediated antitumor immune responses. Cholangiocarcinoma (CCA) has a poor prognosis due its invasiveness and associated metastasis. The present study investigated the expression of B7-H4 in patients with CCA and its association with patient prognosis. The correlation between B7-H4 expression and CD4+ and CD8+ tumor-infiltrating lymphocytes was also investigated. The results demonstrated that high B7-H4 expression was detected in cancer tissues (54/110; 49.1%) compared with that noted in chronic inflammatory bile duct tissues (4/19; 21.1%). Furthermore, all 8 biliary adenoma samples showed negative staining. The expression of B7-H4 was significantly associated with adverse clinical and pathological features including histologic grade (P

The chemical properties and fiber morphology of Dendrocalamus hamiltonii at different age and height were investigated. The variation in SiO2, toluene–alcohol extractive and holocellulose content with culm age was greater than those with height. However, the variation in lignin with culm height was greater than that with age. The fiber morphology of D. hamiltonii differed with age and height, but the differences were not significant. The fiber length, slenderness ratio, and wall thickness reached their greatest values in 2-year growth bamboo culms.

Tea leaf diseases seriously affect the quality and the yield of tea. In order to determine whether the tea leaves are infected by diseases or any types of infection, technical support is essential for taking appropriate measures of disease control. Images of normal tea leaves, tea leaves infected with Tea Red Leaf Spot, and leaves infected with Tea Red Scab disease were studied. An identification algorithm for both of the tea leaf diseases based on hybrid feature optimization was proposed. First, the image features were extracted using the Histogram of Oriented Gradient and the Inception v3 model. Then, hybrid feature optimization processing was performed on two types of extracted features. Finally, the Gradient Boosting Decision Tree algorithm was used as the classifier for the identification of tea leaf diseases. Experiments demonstrate that the hybrid feature optimization algorithm reduces the image feature from 36, 068 to less than 150 dimensions while maintaining a high identification accuracy, which greatly reduces the complexity of the identification algorithm. At the same time, the identification accuracy of tea leaf diseases based on hybrid feature optimization algorithm were higher than 95%.

Sepsis remains the primary cause of death in intensive care units and multiple long non-coding RNAs (lncRNAs) have been demonstrated to be dysregulated in samples of patients with sepsis. However, whether lncRNA-HOTAIR is involved in the etiology of sepsis remains unclear. The aim of the present study was to investigate the role of HOTAIR in sepsis and to reveal the associated mechanisms. A bioinformatics analysis and dual-luciferase reporter assay was performed to evaluate the interaction between HOTAIR and miR-211, as well as miR-211 and IL-6R. An animal model of sepsis was established in mice via cecal ligation and puncture. Interferon (IFN)-γ, interleukin (IL)-6, IL-17, tumor necrosis factor (TNF)-α, IL-1β, IL-6 receptor (R), microRNA (miR)-211 and HOTAIR expression was measured using reverse transcription-quantitative PCR. Cellular proliferation and apoptosis of monocytes were assessed using cell counting kit-8 assay and flow cytometry, respectively. miR-211 was revealed to be targeted by HOTAIR and IL-6R. The expression of IFN-γ, IL-6, IL-17, TNF-α, IL-1β, IL-6R and HOTAIR was significantly upregulated in the septic mice, whereas miR-211 expression was downregulated. The overexpression of hox transcript antisense RNA (HOTAIR) and knockdown of miR-211 were associated with an increased expression of IFN-γ, IL-6, IL-17, TNF-α, IL-1β and IL-6R in monocytes, while the overexpression of miR-211 exhibited the opposite effect. HOTAIR overexpression and miR-211 knockdown significantly inhibited cellular proliferation and promoted monocyte apoptosis, whereas the overexpression of miR-211 exhibited the opposite effects in monocytes. Therefore, HOTAIR may promote the progression of sepsis by indirectly regulating the expression of IL-6R via miR-211.

The aim of this study was to clarify the relationship of nitrogen accumulation and transport with yield ofwheat and determine the indicators for the screening of wheat varieties with high nitrogen use efficiency(NUE). A total of sixteen winter wheat (Triticum aestivum L.) varieties were used to determine fourteennitrogen accumulation and transport-related traits as well as yield-related traits in field under irrigated andrainfed conditions. The correlations between the major nitrogen accumulation and transport-related traitsand yield traits of wheat were analyzed, and the nitrogen use type of the wheat varieties was classified. Resultsshowed that eight nitrogen accumulation and transport-related traits had significant or highly significantgenetic correlations with yield per plant under the two water regimes. These eight traits were transportamount of pre- flowering reserve nitrogen and contribution of pre-flowering reserve nitrogen to grainnitrogen, nitrogen accumulation and transport amount after flowering, nitrogen transport efficiency afterflowering, contribution of nitrogen assimilation to grain nitrogen after flowering, nitrogen utilizationefficiency for grain production and biomass production. These eight traits were used as indicators for acomprehensive clustering of the wheat materials, and the sixteen varieties were classified into three groupsrepresenting the high NUE, intermediate, and low NUE types, respectively. From the perspective of nitrogenutilization, efforts must be made in the following aspects to obtain higher yields regardless of irrigated orrainfed conditions: 1) to improve nitrogen utilization efficiency for grain production and biomass production,as well as nitrogen accumulation and transport amount after flowering, nitrogen transport efficiency andcontribution of nitrogen assimilation to grain nitrogen after flowering, and 2) to decrease transport amount ofpre- flowering reserve nitrogen and contribution of pre-flowering reserve nitrogen to grain nitrogen.

The aim of this study was to determine the neuroprotective effects of walnut protein hydrolysates (WPH) against memory deficits induced by sleep deprivation (SD) in rat and further to identify and characterize the potent neuroprotective peptides against glutamate-induced apoptosis in PC12 cells. Results showed that a remarkable amelioration effect on behavioral performance in Morris water maze test was observed for WPH and its low molecular weight fraction WPHL, especially for WPHL. Additionally, a reduction of antioxidant defense (catalase, glutathione peroxidase (GSH-px), and superoxide dismutase (SOD)) and an increase of malondialdehyde content induced by SD were normalized in brain of rat after oral administration of WPH and WPHL. Then three neuroprotective peptides including GGW, VYY, and LLPF were identified from WPHL, which could protect PC12 cells against glutamate-induced apoptosis with relative cell viability of 78.29 ± 3.09%, 80.65 ± 1.74%, and 83.97 ± 3.06%, respectively, versus glutamate group 48.61 ± 3.99%. The possible mechanism underlying their protective effects of GGW and VYY could be related to their strong radical scavenging activity as well as their ability to reduce reactive oxygen species production and the depletion of SOD and GSH-px in PC12 cells. Notably, the marked neuroprotective effects of LLPF, which did not show obvious free-radical scavenging activity in vitro, could be attributed to its strong effects on inhibiting Ca2+ influx and mitochondrial membrane potential collapse. Additionally, all these peptides could regulate the expression of apoptosis-related proteins (Bax and Bcl-2). Therefore, walnut peptides might be regarded as the potential nutraceuticals against neurodegenerative disorders associated with memory deficits.

With increasing graphene oxide (GO) applications in industry and biomedicine, effects of GO on microorganisms, animals, and human health have been frequently studied; however, direct and indirect effects of GO on plants are seldom concerned. In this study, effects of GO and/or Cd2+ on seed germination, seedling growth, and uptake to Cd2+ were investigated in solution culture. The results showed that GO could quickly adsorb Cd2+ in solution, and the higher the GO concentration was, the lower the residual Cd2+ concentration was in solution. Rice seed germination, seminal root length, and bud length decreased with increasing GO and Cd2+ concentrations respectively, while the presence of GO could alleviate the inhibitive effects of Cd2+ on seminal root and bud growth compared with the single Cd2+ treatment. In maize seedling, fresh weights of shoot and root showed similar responses to the presence of Cd2+ and/or GO. Compared with the single Cd2+ treatment, root Cd concentrations were generally increased by GO in high Cd2+ solution (20 mg/L), while were slightly affected by GO in low Cd2+ solution (5 mg/L) independent of GO concentrations except for 100 mg/L GO. Shoot Cd concentrations were decreased by low GO (100 mg/L) while were increased by high GO (> 500 mg/L) independent of Cd2+ concentrations in solution. Moreover, significant interactive effects of GO and Cd2+ on root and shoot Cd concentrations were observed. This study indicates that GO can change the effects of Cd2+ on seed germination, seedling growth, and uptake to Cd2+ in solution through its adsorption on Cd2+.

In order to increase application of cross-linked enzyme aggregates (CLEAs) in industry production, a novel micro-reactor system that included a rotating magnetic field (RMF), a micro-reactor and CLEAs with magnetic nanoparticles (M-CLEAs) was designed to synthesize butyl oleate. Result showed that the presence of RMF significantly increased the yield of butyl oleate and the maximum increment was 23%. The yield of butyl oleate was impacted by the dosage and distribution of M-CLEAs in micro-reactor. M-CLEAs showed good reusability, since the morphology and the second structure of protein of M-CLEAs did not show evident change after 4 operative cycles. Although the three-dimensional fluorescence of M-CLEAs showed shift in fluorescence intensity and the maximum emission wavelengths, the yield of butyl oleate was not affected. This study provides a novel design that realized efficient, convenient and continuous application of CLEAs in biosynthesis, and M-CLEAs also show good promises in industry production.

Ameliorations of cholinergic system dysfunction and oxidative stress in neurodegenerative diseases were main approaches to improve memory disorder. Our previous investigation showed that anchovy protein hydrolysate (APH) could attenuate scopolamine-induced memory deficits in mice by regulating acetylcholinesterase (AChE) activity. Therefore, peptides with AChE inhibitory activity in APH were explored and identified in this study, and their possible neuroprotective mechanisms on glutamate induced apoptosis in PC12 were also elucidated. Two peptides with strong AChE inhibitory capacity were identified as Pro-Ala-Tyr-Cys-Ser (PAYCS) and Cys-Val-Gly-Ser-Tyr (CVGSY) by ultraperformance liquid chromatography coupled with tandem mass spectrometry. The AChE inhibitory was 23.68 ± 0.97% and 6.08 ± 0.41%, respectively. Treatment with PAYCS and CVGSY could significantly (p < 0.05) increase cells viability, reduce lactate dehydrogenase release, reactive oxygen species (ROS) production, malondialdehyde content, and the ratio of Bax/Bcl-2 of glutamate-induced apoptosis PC12 cells (82.78 ± 6.58 and 109.94 ± 7.16% of control, respectively) as well as increase superoxide dismutase and GSH-px activities. In addition, both the peptides could inhibit Ca2+ influx but have no effects on mitochondrial membrane potential. Results indicated that AChE inhibitory peptides (PAYCS and CVGSY) possibly protected the PC12 cells against glutamate-induced apoptosis via inhibiting ROS production and Ca2+ influx. PAYCS and CVGSY might be considered as nutraceuticals for alleviating memory deficits.

The endogenous soluble sugars and starch are the main carbohydrates accumulated in bamboo culms and the seasonal changes of their contents are also related to the bamboo phenology. In this study, it is observed that the dormancy phase of new Fargesia yunnanensis culms continued as long as 6 months from November to next May. The time interval for the sugar storage between the branching and leafing stage and shooting stage is relatively short. Moreover, the branching and leafing stage overlapped the shoot buds formation stage. Starch granules were depleted when the branching and leafing stage commenced, but then were accumulated before the shooting stage. The endogenous soluble sugar content in 2- and 3-year-old culms decreased during the branching and leafing stage, but increased before the shooting stage, and then decreased after the shooting stage. In shoots, the endogenous soluble sugar and starch contents increased and decreased with culm elongation, respectively. Starch granules were accumulated more in nodes than in internodes in un-elongated internodes. Shoot buds and shoots are the main sink tissues for sugar storage in the whole clumps. The bamboo clumps should be fertilized and irrigated before and after the branching and leafing stage and shooting stage. The late autumn is the best season for the culm harvest.

Suppressor of cytokine signaling 3 (SOCS3) is regarded as a vital repressor in the liver carcinogenesis mainly by inhibiting signal transducer and activator of transcription 3 (STAT3) activity. Farnesoid X Receptor (FXR), highly expressed in liver, has an important role in protecting against hepatocellular carcinoma (HCC). However, it is unclear whether the tumor suppressive activity of FXR involves the regulation of SOCS3. In the present study, we found that activation of FXR by its specific agonist GW4064 in HCC cells inhibited cell growth, induced cell cycle arrest at G1 phase, elevated p21 expression and repressed STAT3 activity. The above anti-tumor effects of FXR were dramatically alleviated by knockdown of SOCS3 with siRNA. Reporter assay revealed that FXR activation enhanced the transcriptional activity of SOCS3 promoter. Electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation (ChIP) assay displayed that FXR directly bound to IR9 DNA motif within SOCS3 promoter region. The in vivo study in nude mice showed that treatment with FXR ligand GW4064 could decelerate the growth of HCC xenografts, up-regulate SOCS3 and p21 expression and inhibit STAT3 phosphorylation in the xenografts. These results suggest that induction of SOCS3 may be a novel mechanism by which FXR exerts its anti-HCC effects, and the FXR-SOCS3 signaling may serve as a new potential target for the prevention/treatment of HCC.

The genetic basis of stomatal conductance (g s), net photosynthetic rate (P N), and transpiration rate (E) was explored by using a wheat doubled haploid population from a cross of Hanxuan10 and Lumai 14. The above three traits were evaluated in wheat flag leaves at 10, 20, 30 days after anthesis under drought stress (DS) and well-watered (WW), and quantitative trait loci (QTL) were analyzed. Expression of the traits during the grain filling stage showed downward trends under both conditions, but expression of three phenotypes were stronger under WW than those under DS. Extremely significant positive correlations were established among the traits at all growth stages under both conditions. A total of 18 additive QTLs for those traits were identified on 10 chromosomes. Among them, two batches of nine additive QTLs were associated with the target traits under DS and WW, respectively. Two additive QTLs for g s and E, two for g s and P N, six for g s, P N, and E clustered at the same or near the region (colocation) of chromosomes 4A, 2B, and 7B, respectively. This provided genetic basis for close phenotype correlations among g s, P N, and E. Furthermore, QTLs for g s, P N, and E near Xgwm577 and Xgwm611 located on 7B chromosome were linked to previously reported QTLs regulating a SPAD value and the chlorophyll a/b ratio under dark-induced condition. This finding indicated that these QTLs on 7B chromosome might be involved in the process of wheat leaf senescence.

To explore the expression of lamininγ2 in extrahepatic cholangiocarcinoma (EHCC) tissues and its influence on tumor invasion and metastasis.Paraffin embedding samples of cancer, para-cancer, lymph node metastatic and hepatic metastatic tissues from 79 patients undergoing EHCC resection were collected. Expression of lamininγ2 was detected by immunohistochemistry and its relationship with clinical pathological characteristics and the prognosis of EHCC patients were analyzed.Lamininγ2 showed negative staining in para-cancer tissues, but demonstrated a 51.9% (41/79) positive expression rate in extracellular matrix (ECM) or cytoplasm of EHCC tissues. In lymph node metastatic and distant metastatic nidi, expression of lamininγ2 was significantly higher than in the primary nidi (χ2=7.4173, P=0.0065; χ2=4.0077, P=0.0453). The expression was in obvious association with lymph node metastasis (P0.01), but had no relevance with age, gender, tumor location, tumor stage, differentiation and distant metastasis in ECM (P0.05), whereas it was in marked connection with lymph node and distant metastasis (P0.05 or P0.01), but had no relationship with age, gender, tumor location, tumor stage and differentiation in cytoplasm (P0.05). However, the median survival time and median recurrent period of patients with positive expression of lamininγ2 in both cytoplasm and ECM of tumor cells, only in ECM and only in cytoplasm, were evidently lower than with negative expression of lamininγ2 in RCM and cytoplasm (P0.05 or P0.01). Further Cox regression analysis showed that the positive expression of lamininγ2 and the tumor differentiation were independent risk factors influencing the prognosis of EHCC patients.Abnormal expression of lamininγ2 may be closely associated with invasion and metastasis of tumor cells, and thus a potential molecular marker for prognosis of EHCC patients.

Nitrogen-doped multichanneled activated carbon (NMAC) has been prepared for application as electrocatalysts for oxygen reduction reaction (ORR) in fuel cells. The NMAC is prepared by carbonizing luffa sponge fibers in NH 3 and subsequent activation with KOH. The NMAC possesses densely packed and parallel channels with diameters of 3–13 μm and wall thickness of 0.4–1.8 μm. After activation micro- and mesopores are generated on the channel walls, forming a hierarchically porous structure. The NMAC exhibits excellent electrocatalytic performance for ORR both in alkaline and acidic media with high electrocatalytic activity and long-term operation stability. In particular, the NMAC has prominent stability in acidic media, keeping stable after about 7 hours and retaining 88.5% of the initial current density after 12 hours in 0.5 M H 2 SO 4 solution. Because the active sites are located on the inner surface of the micrometer-scale channels the performance degradation caused by peeling, aggregation, and crack during operation can be avoided, resulting the high stability. Considering the abundance of luffa fibers on earth, the present NMAC materials are highly promising in various applications such as electrocatalysts, supercapacitors, and templates for supporting various functional materials.

Culm sheath has always been considered a modified leaf. In this study, the anatomical structure of the culm sheath from Fargesia yunnanensis was analyzed to determine whether it originated from the foliage leaf blade or from the branch. The vascular bundles of the culm sheath showed greater similarity to the branches in shape and anatomical structure. In contrast to foliage blades, there are no midribs in culm sheaths. Stomatal density in the culm sheath is greater in the adaxial than it is in the abaxial epidermis, which is the opposite of that found in foliage leaf blades, and that density shows greater similarity to branches than it does to foliage blades. Fusoid cells are distributed on both sites of the vascular bundles in foliage blades, whereas culm sheaths have, instead, a few parenchyma cells that disintegrate and form air cavities when the sheath matures. Additionally, the culm sheath has no bulliform cells or trichome in its epidermis, and the shape of its long cells shows greater similarity to those of branches. Therefore, culm sheath is a modified branch, rather than a modified leaf.

SUMMARYDelayed senescence, or stay-green, contributes to a longer grain-filling period and has been regarded as a desirable characteristic for the production of a number of crops including wheat. In the present study, in order to identify quantitative trait loci (QTLs) for traits related to the progression of wheat flag leaf senescence, green leaf area duration (GLAD) of a doubled haploid (DH) population, derived from two winter wheat varieties Hanxuan10 and Lumai14, was visually estimated under two water conditions and was recorded at 3-day intervals from 10 days after anthesis to physiological maturity using a 0–9 scale. According to GLAD, parameters related to the progression of senescence of DH lines and their parents were estimated by the Gompertz statistical model. Based on the model parameters, DH lines were categorized into three groups under drought stress and four groups under well-watered conditions. A total of 24 additive QTLs and 23 pairs of epistatic QTLs for parameters related to the progression of senescence were identified on 18 chromosomes, except for 3B, 1D and 6D. Of the QTLs detected, 14 and 10 additive QTLs were associated with the investigated traits under drought stress and well-watered conditions, respectively. Furthermore, 4, 7, 6, 2 and 2 additive QTLs for traits related to progression of senescence were clustered around the same or similar regions of chromosomes 1A, 1B, 5A, 5B and 7A, respectively. The present data provided the genetic basis for high phenotypic correlations among traits related to the progression of wheat flag leaf senescence. In addition, 17 loci were co-located or linked with previously reported QTLs regulating chlorophyll fluorescence, high-light-induced photo-oxidation, or heat stress and dark-induced senescence. The markerXwmc336on chromosome 1A, responsible for the onset and end times of leaf senescence, the time to maximum rate of senescence, the time to reach 75% senescence and chlorophyll content under drought stress may be helpful for marker-assisted selection breeding of wheat.

Elevated ozone (O3) generally affects microbial biomass and community structure in rhizosphere, but these effects are unclear in mycorrhizal plants because arbuscular mycorrhizal (AM) fungi often benefit microbial growth in the rhizosphere. Here, we investigate the effects of elevated O3 on microbial biomass and community structure in the rhizosphere of mycorrhizal snap bean (Phaseolus vulgaris L.) with different O3 sensitivity (R123: O3-tolerant plant; S156: O3-sensitive plant) based on the phospholipid fatty acids (PLFAs) method. Compared with ambient O3, elevated O3 significantly decreased mycorrhizal colonization rates in the 2 genotypes, especially in S156 plants. The wet masses of shoot and root were decreased by elevated O3 in the 2 genotypes independent of AM inoculation, but they were higher in the mycorrhizal plant than in the nonmycorrhizal plant independent of O3 concentration. Elevated O3 significantly decreased the relative proportion of specific fungal PLFAs in the nonmycorrhizal plant, but this effect disappeared in the mycorrhizal plant. The relative proportions of specific PLFAs of other microbial groups (Gram-positive, Gram-negative, and actinomycete) in the rhizosphere and all specific PLFAs in the hyphosphere were not affected by elevated O3 independent of AM inoculation. In the rhizosphere of the 2 genotypes, microbial community structure was changed by AM inoculation and elevated O3 as well as by their interaction; in the hyphosphere, however, microbial community structure was changed by elevated O3 only in R123 plants. It is concluded that AM inoculation can offset negative effect of elevated O3 on fungal biomass but seems to enhance shift of microbial community structure in rhizosphere under elevated O3.