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Jun Ye,1,2 Renjie Li,1,2 Jialing Cheng,1,2 Dongdong Liu,1,2 Yanfang Yang,1,2 Hongliang Wang,1,2 Xiaoyan Xu,1,2 Lin Li,1,2 Panpan Ma,3 Yuling Liu1,2 1State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, People’s Republic of China; 2Beijing Key Laboratory of Drug Delivery Technology and Novel Formulation, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, People’s Republic of China; 3Beijing Union Second Pharmaceutical Factory, Beijing, People’s Republic of ChinaCorrespondence: Yuling Liu, State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, 1 Xiannongtan Street, Beijing, 100050, People’s Republic of China, Tel +86 10-89285188, Fax +86 10-89285190, Email ylliu@imm.ac.cnBackground: Amphotericin B (AmB) nanoformulations have been widely used for the treatment of invasive fungal infections in clinical practice, all of which are lyophilized solid dosage forms that improve storage stability. The colloidal stability of reconstituted lyophilized nanoparticles in an injection medium is a critical quality attribute that directly affects their safety and efficacy during clinical use.Methods: In the present study, the colloidal stability of commercial AmB nanoformulations, including AmB cholesteryl sulfate complex (AmB-CSC) and AmB liposome (AmB-Lipo), was evaluated using the dynamic (DLS) and static multiple light scattering (SMLS) techniques.Results: Compared to the DLS technique, the SMLS technique allows for a more objective and accurate evaluation of the colloidal stability of AmB nanoformulations. The results obtained using the SMLS technique demonstrated that AmB-CSC and AmB-Lipo exhibited excellent colloidal stability in both sterile water and 5% dextrose injection. The disk-like structure of the AmB-CSC nanoparticles more readily adsorbed serum proteins to form protein corona compared to the spherical structure of AmB-Lipo after incubation with serum. Additionally, AmB-CSC and AmB-Lipo can significantly reduce the in vitro cytotoxicity and in vivo nephrotoxicity of AmB, which may be attributed to the good colloidal stability and the improved pharmacokinetic profiles of AmB nanoformulations.Conclusion: To the best of our knowledge, this study is the first to compare the colloidal stability of commercial AmB nanoformulations. These findings will provide useful information not only to inform the clinical use of available AmB nanoformulations but also for improving the design and conduct of translational research on novel AmB nanomedicines.Keywords: nano-drug delivery systems, colloidal stability, amphotericin B, dynamic light scattering, static multiple light scattering

The first HFIP-promoted catalyst-free cascade reactions for the synthesis of biologically relevant 3,3-di(indolyl)indolin-2-ones (27 examples, up to 98% yield) from readily available indoles and isatin derivatives are described. This protocol shows well tolerance of different functional groups and features extremely mild reaction conditions such as short reaction time (∼ 1 h), no usage of catalyst, easy operation and product isolation. Of particular interest is the formation of two C-C bonds and one all-carbon quaternary center. This protocol could serve as an alternative strategy to synthesize biologically important 3,3-di(indolyl)indolin-2-ones for biological testing.

Targeting bromodomain-containing protein 4 (BRD4) has been proved to be an effective strategy for cancer therapy. To date, numerous BRD4 inhibitors and degraders have been identified, some of which have advanced into clinical trials. In this work, a focused library of new [1,2,4]triazolo[1,5-a]pyrimidine derivatives were discovered to be able to inhibit BRD4. WS-722 inactivated BRD4 (BD1/BD2), BRD2 (BD1/BD2) and BRD3 (BD1/BD2) broadly with the IC50 values less than 5 μmol/L. Besides, WS-722 inhibited growth of THP-1 cells with an IC50 value of 3.86 μmol/L. Like (+)-JQ1, WS-722 inhibited BRD4 in a reversible manner and enhanced protein stability. Docking studies showed that WS-722 occupied the central acetyl-lysine (Kac) binding cavity and formed a hydrogen bond with Asn140. In THP-1 cells, WS-722 showed target engagement to BRD4. Cellular effects of WS-722 on THP-1 cells were also examined, showing that WS-722 could block c-MYC expression, induce G0/G1 phase arrest and p21 up-regulation, and promote differentiation of THP-1 cells. BRD4 inhibition by WS-722 resulted in cell apoptosis and up-regulated expression of cleaved caspased-3/7 and PARP in THP-1 cell lines. The [1,2,4]triazolo[1,5-a]pyrimidine is a new template for the development of new BRD4 inhibitors.

The fast growth of mulberry depends on high water consumption, but considerable variations in drought tolerance exist across different cultivars. Physiological and anatomical mechanisms are important to plant survival under drought. However, few research efforts have been made to reveal the relationships of these two aspects in relation to drought tolerance. In this study, growth rates, leaf functional physiology and anatomical characteristics of leaf and xylem of 1-year-old saplings of seven mulberry cultivars at a common garden were compared. Their relationships were also explored. Growth, leaf physiology and anatomy were significantly different among the tested cultivars. Foliar stable carbon isotope composition (δ13 C) was negatively correlated with growth rates, and closely related to several leaf and xylem anatomical traits. Particularly, leaf thickness, predicted hydraulic conductivity and vessel element length jointly contributed 77% of the variability in δ13 C. Cultivar Wupu had small stomata, intermediate leaf thickness, the smallest hydraulically weighted vessel diameter and highest vessel number, and higher δ13 C; Yunguo1 had high abaxial stomatal density, low specific leaf area, moderate hydraulic conductivity and δ13 C; these are beneficial features to reduce leaf water loss and drought-induced xylem embolism in arid areas. Cultivar Liaolu11 had contrasting physiological and anatomical traits compared with the previous two cultivars, suggesting that it might be sensitive to drought. Our findings indicate that growth and δ13 C are closely associated with both leaf and xylem anatomical characteristics in mulberry, which provides fundamental information to assist evaluation of drought tolerance in mulberry cultivars and in other woody trees.

Drought stress commonly happens more than once during the life cycle of perennial trees. Stress memory endows better capacity to cope with repeated stresses for plants, while the underlying mechanisms are not fully elucidated. In this study, 2-month-old saplings of two mulberry cultivars (Husang32 and 7307 of Morus multicaulis) with or without an early soil water deficit were subjected to subsequent drought for 9 days. The shoot height growth, biomass production, stable carbon isotope discrimination, phytohormones, reactive oxygen species (ROS), osmotic substances and antioxidant enzymes were analyzed after the first and the second drought, respectively. Drought priming saplings sustained comparable or slightly higher biomass accumulation under the second drought than those non-priming. They also exhibited decreased levels of soluble sugars, free proline and soluble proteins, lower accumulation of malonaldehyde (MDA) and superoxide anion (O

Auxin plays essential roles in plant normal growth and development. The auxin signaling pathway relies on the auxin gradient within tissues and cells, which is facilitated by both local auxin biosynthesis and polar auxin transport (PAT). The TRYPTOPHAN AMINOTRANSFERASE OF ARABIDOPSIS (TAA)/YUCCA (YUC) pathway is the most important and well-characterized pathway that plants deploy to produce auxin. YUCs function as flavin-containing monooxygenases (FMO) catalyzing the rate-limiting irreversible oxidative decarboxylation of indole-3-pyruvate acid (IPyA) to form indole-3-acetic acid (IAA). The spatiotemporal dynamic expression of different YUC gene members finely tunes the local auxin biosynthesis in plants, which contributes to plant development as well as environmental responses. In this review, the recent advances in the identification, evolution, molecular structures, and functions in plant development and stress response regarding the YUC gene family are addressed.

The initiation and induction of root primordia are of great importance for adventitious root (AR) formation in cutting propagation of horticultural and forestry crops. However, the underlying mechanisms orchestrating these early phases of AR formation remain largely unexplored. Here, we investigated the physiological and transcriptomic changes during the early AR phases in mulberry stem hardwood cuttings. The results showed that the concentrations of soluble proteins increased, whereas concentrations of soluble sugars and starch were decreased. Indole-3-acetic acid (IAA) and zeatin had a rapid transit peak at 6 h after planting (hAP) and declined thereafter. The activities of peroxidase and catalase persistently increased and indole-3-acetic acid oxidase was maintained at a higher stable level from 0 hAP, while the activities of polyphenol oxidase fluctuated with soluble phenolics and IAA levels. The comparative transcriptome identified 4276 common genes that were differentially regulated at −6, 0 and 54 hAP. They were separated into five clusters with distinct biological functions such as defense response and photosynthesis. Considerable common genes were assigned to pathways of sugar metabolism, mitogen-activated protein kinase, and circadian rhythm. The gene co-expression network analysis revealed three major co-expressed modules involved in stress responses, hormone signaling, energy metabolism, starch metabolism, and circadian rhythm. These findings demonstrate the positive effect of auxin on AR induction, and uncovered the crucial roles of stress responses, hormone signaling and circadian rhythm in coordinating the physiological changes during the early phases of AR formation in mulberry stem hardwood cuttings.

Physiological and morphological traits have a considerable impact on the biomass production of fast-growing trees. To compare cultivar difference in shoot biomass and investigate its relationships with leaf functional traits in mulberry, agronomic traits and 20 physiological and morphological attributes of 3-year-old mulberry trees from eight cultivars growing in a common garden were analyzed. The cultivars Xiang7920, Yu711, and Yunsang2 had higher shoot fresh biomass (SFB), which was closely associated with their rapid leaf expansion rate, large leaf area, and high stable carbon isotope composition (&delta, 13C). Conversely, the cultivars 7307, Husang32, Wupu, Yunguo1, and Liaolu11 were less productive, and this was primarily the result of slower leaf expansion and smaller leaf size. Growth performance was negatively correlated with leaf &delta, 13C and positively correlated with the total nitrogen concentration, indicating that a compromise exists in mulberry between water use efficiency (WUE) (low &delta, 13C) and high nitrogen consumption for rapid growth. Several morphological traits, including the maximum leaf area (LAmax), leaf width and length, petiole width and length, leaf number per shoot, and final shoot height were correlated with SFB. The physiological traits that were also influential factors of shoot biomass were the leaf &delta, 13C, the total nitrogen concentration, and the water content. Among the studied leaf traits, LAmax, leaf &delta, 13C, and concentrations of chlorophyll a and b were identified as the most representative predictor variables for SFB, accounting for 73% of the variability in SFB. In conclusion, a combination of LAmax, leaf &delta, 13C, and chlorophyll should be considered in selection programs for high-yield mulberry cultivars.

The ability to form mature adventitious roots (AR) provides a competitive advantage for clonal multiplication of elite genotypic plant species, because high economic losses occur as a result of insufficient rooting. To better understand potential mechanisms involved in AR formation, we utilized an iTRAQ-based proteomic approach on mulberry hardwood cuttings. A total of 4427 proteins were identified from the base of cuttings, of which 595 and 231 proteins showed differential accumulations in the two periods of rooting, respectively. Three differentially expressed enzyme proteins were validated by an enzyme assay and qPCR. Functional annotation analysis showed that dysregulated proteins were involved in glucose metabolism, flavonoids biosynthesis, cell wall modification, and hormone regulation, indicating potential contributions to adventitious rooting. These results provide fundamental and important information for research on the molecular mechanism of AR development in mulberry cuttings and facilitate rooting efficiency in agricultural practice.

Adventitious rooting derived from the parenchyma cells of vascular cambium is a highly efficient rooting type for cutting propagation of woody species. To unravel the molecular mechanisms underlying this distinct rooting process in mulberry stem hardwood cuttings, RNA-seq approach was adopted to perform wide transcriptional expression profiles of three stages characterized by initial excision (stage 1), root primordia induction (stage 2) and emergence of adventitious roots (stage 3). Compared to the former two stages, stage 3 had the most expressed genes (19,987) to establish functionally mature adventitious roots. In contrast, differentially expressed genes (DEGs) identified in stage 1 vs stage 2 comparison group were over threefold higher than stage 2 vs stage 3 comparison group (8930 vs 2739), suggesting a drastic transcriptional activation of genes involved in root primordia induction. GO and KEGG enrichment analysis of the DEGs showed that circadian rhythm-plant pathway was significantly enriched in response to environmental and endogenous changes in stage 2, whereas plant hormones including auxin, ethylene and cytokinin coordinately participated in the formation of adventitious roots in a stage-specific manner via an extensive and delicate regulation of their biosynthesis and signal transduction. Among which, several sets of genes relevant to circadian clock and hormone biosynthesis and signaling are speculated to potentially associated with the induction and differentiation of parenchyma cell initiated adventitious rooting. These findings will enrich our understandings to the mechanisms of adventitious rooting in mulberry, and provide theoretical support for the development of rooting techniques for other woody crops.

Due to the non-standardization of online comments, it is difficult to extract the "opinion target - opinion word" pairs. This paper proposes a method of extracting the "opinion target - opinion word" pairs based on part-of-speech rules and semantic dependency parsing (SDP). Firstly, adjectives, status words and some idioms are extracted as original opinion words according to the part-of-speech characteristics of opinion words, and then SDP is performed to obtain the final opinion words. Then, the opinion targets are extracted based on SDP and dependency parsing (DP), and filtered through part-of-speech rules. Finally, the implicit opinion targets are extracted based on probability. The experimental results show that the average F-Measure is 78.86%, which indicates that the extraction performance of this method is good.

Accelerated root formation is necessary for successful cuttage in a number of woody plants, such as mulberry (Morus alba L.). However, little information regarding the molecular mechanisms involved in this type of root formation is available. Here, we compared transcriptome changes during different root formation periods (0, 4, and 8 days) by conducting high-throughput sequencing to understand the molecular mechanisms of root formation in mulberry softwood cuttings. Approximately 88.2 million, 88.2 million, and 52.8 million transcriptome sequencing reads were obtained for the three time periods, 0, 4, and 8 days, respectively. These reads were then assembled into 61,687 (0 day), 61,651 (4 days), and 59,457 (8 days) contigs by utilizing the de novo method, and a large number of differentially expressed genes were identified at various developmental stages. These assembled contigs were annotated using Kyoto Encyclopedia of Genes and Genomes (KEGG), as well as the simple sequence repeat (SSR). Mulberry had a complex response to adventitious rooting. Genes that might be associated with mulberry stem cutting root formation were also identified here. Furthermore, the reliability of the sequencing results was validated by performing quantitative real-time polymerase chain reaction (qRT-PCR). These results provide a comprehensive background of the molecular biology of mulberry development, in particular regarding its stem cutting root formation development.