QTL Mapping of Agronomic and Physiological Traits at the Seedling and Maturity Stages under Different Nitrogen Treatments in Barley.

Nitrogen (N) stress seriously constrains barley (Hordeum vulgare L.) production globally by influencing its growth and development. In this study, we used a recombinant inbred line (RIL) population of 121 crosses between the variety Baudin and the wild barley accession CN4027 to detect QTL for 27 traits at the seedling stage in hydroponic culture trials and 12 traits at the maturity stage in field trials both under two N treatments, aiming to uncover favorable alleles for N tolerance in wild barley. In total, eight stable QTL and seven QTL clusters were detected. Among them, the stable QTL Qtgw.sau-2H located in a 0.46 cM interval on the chromosome arm 2HL was a novel QTL specific for low N. Notably, Clusters C4 and C7 contained QTL for traits at both the seedling and maturity stages. In addition, four stable QTLs in Cluster C4 were identified. Furthermore, a gene (HORVU2Hr1G080990.1) related to grain protein in the interval of Qtgw.sau-2H was predicted. Correlation analysis and QTL mapping showed that different N treatments significantly affected agronomic and physiological traits at the seedling and maturity stages. These results provide valuable information for understanding N tolerance as well as breeding and utilizing the loci of interest in barley.

Multiple-Enzyme Graphene Microparticle Presenting Adaptive Chemical Network Capabilities.

Interrelated reaction networks steered by multiple types of enzymes are among the most intriguing enzyme-based cellular features. These reaction networks display advanced features such as adaptation, stimuli-responsiveness, and decision-making in accordance with environmental cues. However, artificial enzyme particles are still deficient in network-level capabilities, mostly because delicate enzymes are difficult to immobilize and assemble. In this study, we propose a general strategy to prepare enzyme-based particles that demonstrate network reaction capability. We assembled multiple types of proteins with a nanoscopic binder prepared from polyelectrolyte and graphene. After assembly, the enzymes all preserved their catalytic capabilities. By incorporating multiple types of enzymes, the particles additionally displayed network-reaction capabilities. We were able to use NIR irradiations to quasi-reversibly adjust the catalytic abilities of these enzyme-based particles. In addition, after a biomimetic mineralization process was used to wrap the protein complexes in a MOF shell, the particles were more robust and catalytically active even after being immersed in acidic (pH 4) or basic (pH 10) solutions for 3 days. This study provides an insight into the study of network properties of functional enzyme particles experimentally and enriches scientific understanding of multifunctional or stimuli-responsive behaviors at the reaction network level. The building of artificial reaction networks possesses high potential in realizing intelligent microparticles that can perform complicated tasks.

Metabolomics Study of Cultivated Bulbus Fritillariae Cirrhosae at Different Growth Stages using UHPLC-QTOF-MS Coupled with Multivariate Data Analysis.

INTRODUCTION: Bulbus fritillariae cirrhosae (known as Chuan bèi mǔ in China, BFC) contain fritillaria steroidal alkaloids as the bioactive ingredients and are widely used as traditional Chinese medicine for the treatment of cough and phlegm. Due to limited wild resources, the cultivated species are becoming predominantly used in Chinese traditional medicine markets. OBJECTIVE: To assess the impact of different growth stages on the alkaloids of cultivated BFC and establish a reference for quality control and guidance for appropriate harvesting practices. METHODS: The ultra-high performance liquid chromatography quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS) metabolomic strategy was applied to determine potential chemical markers for the discrimination and quality control of cultivated BFC in different growth stages. The molecular feature extraction and multivariate statistical analysis were applied to alkaloid extraction and full metabolomic profiling of cultivated BFC for classification and marker compound characterisation. RESULT: This approach allowed the establishment of a fast and efficient comparative multivariate analysis of the metabolite composition of 42 samples covering growth of cultivated BFC ranging in age from one to seven years old. Four alkaloid compounds were identified in cultivated BFC based on accurate mass, retention time, and MS/MS fragments. These compounds may be used as potential chemical markers for the classification and discrimination of cultivated BFC samples indifferent growth stages. CONCLUSIONS: The proposed analytical method in combination with multivariate statistical analysis comprised a useful and powerful strategy to explore the chemical ingredients and transforming mechanisms of cultivated BFC and for quality evaluation and control. Copyright © 2018 John Wiley & Sons, Ltd.

The use of 5-fluorouracil-loaded nanobubbles combined with low-frequency ultrasound to treat hepatocellular carcinoma in nude mice.

OBJECTIVE: This study aimed to investigate the therapeutic effects of 5-fluorouracil (5-FU)-loaded nanobubbles irradiated with low-intensity, low-frequency ultrasound in nude mice with hepatocellular carcinoma (HCC). METHODS: A transplanted tumor model of HCC in nude mice was established in 40 mice, which were then randomly divided equally into four groups: group A (saline), group B (5-FU-loaded nanobubbles), group C (5-FU-loaded nanobubbles with non-low-frequency ultrasound), and group D (5-FU-loaded nanobubbles with low-frequency ultrasound). The tumor size in each mouse was observed via ultrasound before and after the treatments. Inhibition of the tumor growth in each group was compared, and survival curves were generated. Tumor tissues were removed to determine the apoptotic index using the TUNEL method and quantitative analysis. Tumor tissues with CD34-positive microvessels were observed by immunohistochemistry, and the tumor microvessel densities were calculated. RESULTS: The growth rate of the tumor volumes in group D was significantly slower than that in the other groups, while the tumor inhibition rates and apoptotic index in group D were significantly higher than those of the other groups. The number of microvessels staining positive for CD34 was decreased in group D. Therefore, group D presented the most significant inhibitory effects. CONCLUSIONS: Therefore, 5-FU-loaded nanobubbles subjected to irradiation with low-frequency ultrasound could further improve drug targeting and effectively inhibit the growth of transplanted tumors, which is expected to become an ideal drug carrier and targeted drug delivery system for the treatment of HCC in the future.

Upregulation of Bcl-2 enhances secretion of growth factors by adipose-derived stem cells deprived of oxygen and glucose.

There is an increasing recognition that beneficial effects of adipose-derived stem cell (ADSC) therapy may depend largely on the secretion of multiple growth factors. This study modified ADSCs with the Bcl-2 gene in order to increase the secretion of growth factors during oxygen-glucose deprivation (OGD). The phenotypes of human ADSCs that were passaged 4 times were analyzed using flow cytometry. Then, ADSCs were genetically modified with Bcl-2 and Bcl-2 gene transduction was verified with Western blotting. Proliferative capacity and multipotent differentiation properties were evaluated in Bcl-2-modified ADSCs. Secretion of vascular endothelial growth factor (VEGF), hepatocyte growth factor (HGF), and basic fibroblast growth factor (bFGF) was evaluated using an enzyme-linked immunosorbent assay (ELISA) during OGD. Human ADSCs that were passaged 4 times expressed stem cell-associated markers but not a fibroblast marker or a hematopoietic stem cell marker. The Bcl-2 gene was efficiently transfected into ADSCs; Bcl-2 modification did not affect the proliferative and multilineage differentiation capacity of ADSCs. In addition, Bcl-2 overexpression enhanced the secretion of VEGF, bFGF, and HGF by 14.47%, 16.9%, and 91%, respectively, compared to ADSCs alone that were deprived of oxygen and glucose. These data suggest that Bcl-2 overexpression enhances secretion of angiogenic growth factors by ADSCs deprived of oxygen and glucose.

Penthorin A and B, two unusual 2,4'-epoxy-8,5'-neolignans from Penthorum chinese.

Two new 2,4'-epoxy-8,5'-neolignans (1 and 2), together with five known 7,9';7',9-diepoxylignans (3-7), were isolated from an ethyl acetate soluble portion of a hepatoprotective water decoction of Penthorum chinese. Their structures and absolute configurations were elucidated by extensive spectroscopic analyses and electronic circular dichroism (ECD) calculations. This is the second report of 2,4'-epoxy-8,5'-neolignans from plants. Compounds 2, 6, and 7 showed in vitro protective activities against acetaminophen-induced hepatocyte injury at 5 µM.

New amino butenolides from the bulbs of Fritillaria unibracteata.

Five new amino γ-butenolides, fritenolide A (1), B (2), C (3), D (4), and E (5), along with four known compounds, were isolated from the bulbs of Fritillaria unibracteata. Their structures were determined by spectroscopic analysis, including 1D NMR, 2D NMR, HRESIMS, HRESIMS/MS, IR, and CD techniques. All isolates were evaluated for the protective activity on injured hepatocytes and cytotoxic activity on human cancer cells in vitro. The unusual amino butenolides were isolated from the Liliaceae family for the first time.

Chemical composition and antibacterial activity of essential oils from different parts of Leonurus japonicus Houtt.

The herb and fruits of Leonurus japonicus Houtt., named "Yimucao" and "Chongweizi", respectively, in Chinese, have been widely used in China as gynecological medicines. The components of the essential oils obtained by hydrodistillation were investigated by GC-MS. The antibacterial activity of the essential oils was determined by micro-dilution assay. The results showed large variations in the chemical composition and antibacterial activity of the oils. The oil of "Yimucao" showed antibacterial activity against various Gram-positive bacteria and consisted mainly of sesquiterpenes and diterpenes, with phytone, phytol, caryophyllene oxide and ß-caryophyllene being the most significant constituents, whereas the oil of "Chongweizi", mainly made up of bornyl acetate and aliphatic hydrocarbons, was inactive in the antibacterial assay. Further study of the main compounds in "Yimucao oil" showed that ß-caryophyllene had wide-spectrum activity against Gram-positive bacteria.

Alkaloids isolated from the lateral root of Aconitum carmichaelii.

Two new alkaloids, aconicarmine (1) and aconicaramide (5), were isolated from the EtOH extract of the lateral roots of Aconitum carmichaelii, together with five known compounds: fuziline (2), neoline (3), N-ethylhokbusine B (4), 5-hydroxymethylpyrrole-2-carbaldehyde (6), and oleracein E (7). Their structures were elucidated by physical and NMR analysis. Pyrrole alkaloids were isolated from A. carmichaelii for the first time. In the in vitro assays, compounds 2 and 3 showed activity against pentobarbital sodiuminduced cardiomyocytes damage by obviously recovering beating rhythm and increasing the cell viability, while compounds 5 and 7 showed moderate antibacterial activity.