Discovery of potent small molecule inhibitors of histone lysine methyltransferase NSDs.

Nuclear receptor binding SET domain (NSD) proteins are a class of histone lysine methyltransferases and implicated in multiple cancer types with aberrant expression and involvement of cancer related signaling pathways. In this study, a series of small-molecule compounds including compound 2 and 3 are identified against the SET domain of NSDs through structure-based virtual screening. Our lead compound 3 exhibits potent inhibitory activities in vitro towards the NSD2-SET and NSD3-SET with an IC50 of 0.81 µM and 0.84 µM, respectively, and efficiently inhibits histone H3 lysine 36 dimethylation and decreases the expression of NSDs-targeted genes in non-small cell lung cancer cells at 100 nM. Compound 3 suppresses cell proliferation and reduces the clonogenicity in H460 and H1299 non-small cell lung cancer cells, and induces s-phase cell cycle arrest and apoptosis. These data establish our compounds as a valuable tool-kit for the study of the biological roles of NSDs in cancer.

Quantifying RNA structures and interactions with a unified reduced chain representation model.

RNA is a pivotal molecule that plays critical roles in various cellular processes. Quantifying RNA structures and interactions is essential to understanding RNA function and developing RNA-based therapeutics. Using a unified five-bead model and a non-redundant database, this paper investigates the structural features and interactions of five commonly occurring RNA motifs, i.e., double-stranded helices, hairpin loops, internal/bulge loops, multi-branched junctions, and single-stranded terminal tails. Analyzing detailed distributions of RNA local structural features and base-base interactions reveals a preference for helical structures in both local backbone structures and base orientations. The interactions between adjacent bases exhibit motif-specific and sequence-dependent characteristics, reflecting the distinct topological constraints imposed by different loop-helix connection modes and the varying pairing and stacking interactions among different sequences. These findings shed light on the stability of RNA helices, emphasizing their significance in providing dominant base pairing and stacking interactions for RNA structures and stability. The four non-helix motifs encompass unpaired nucleotide loops and exhibit diverse base-base interactions, contributing to the structural diversity observed in RNA. Overall, the complexity of RNA structure arises from the intricate interplay of base-base interactions.

A molecular understanding of citrate adsorption on calcium oxalate polyhydrates.

Calcium oxalate precipitation is a common pathological calcification in the human body, whereby crystallite morphology is influenced by the chelating properties of biological ions such as citrate. It has been suggested that citrate could steer oxalate formation towards its dihydrated form and away from the monohydrated form, which was identified as a major cause for disease. To assess the influence of the citrate ion on the resulting calcium oxalate, surface energies were calculated at the dispersion-corrected density functional level of theory for both monohydrated and dihydrated calcium oxalate. Different adsorption geometries were considered by varying the attacking angle of citrate as well as by considering the citrate ion on top of an adsorbed water layer or penetrating the water layer. The obtained results were compared to ab initio molecular dynamics simulations and experimental scanning electron microscope images. A strong preference for citrate adsorption on calcium oxalate dihydrate was observed, suggesting medical applications for the treatment of such pathological calcifications.

Porous BCP ceramics with nanoscale whisker structure accelerate bone regeneration by regulating inflammatory response.

Inflammation-induced by biomaterials is a critical event to determine the success and efficiency of tissue repair. Macrophages are a major population that participates the biomaterial induced inflammation. The response of macrophages depends on the characteristics of biomaterials, thus causing a cascade reaction in subsequent biological processes. In this study, porous biphase calcium phosphate (BCP) ceramics with the different surface structures were constructed to compare the effect of surface structure on bone generation potential, and further reveal the inflammation-involved mechanism. Our results demonstrated that macrophages on three ceramics showed distinct morphologies and spreading areas. The nanoscale whisker structure did induce more bone generation in the mice thigh muscle. The in vitro result revealed that nanoscale whisker structure could drive macrophage polarization towards M1-like phenotype, indicated by a higher expression of pro-inflammatory specific markers (iNOS and CCR7), and mass secretion of TNF-α. Further research indicated that additional TNF-α could promote the osteogenic differentiation of mesenchymal stem cells (MSCs). However, excess addition of TNF-α showed an opposite effect on the osteogenic differentiation of MSCs by initiating the NF-κB signaling pathway, which suppresses the osteogenesis process.

Intracellular infection by symbiotic bacteria requires the mitotic kinase AURORA1.

The subcellular events occurring in cells of legume plants as they form transcellular symbiotic-infection structures have been compared with those occurring in premitotic cells. Here, we demonstrate that Aurora kinase 1 (AUR1), a highly conserved mitotic regulator, is required for intracellular infection by rhizobia in Medicago truncatula. AUR1 interacts with microtubule-associated proteins of the TPXL and MAP65 families, which, respectively, activate and are phosphorylated by AUR1, and localizes with them within preinfection structures. MYB3R1, a rhizobia-induced mitotic transcription factor, directly regulates AUR1 through two closely spaced, mitosis-specific activator cis elements. Our data are consistent with a model in which the MYB3R1-AUR1 regulatory module serves to properly orient preinfection structures to direct the transcellular deposition of cell wall material for the growing infection thread, analogous to its role in cell plate formation. Our findings indicate that the eukaryotically conserved MYB3R1-TPXL-AUR1-MAP65 mitotic module was conscripted to support endosymbiotic infection in legumes.

Crystalline structures of L-cysteine and L-cystine: a combined theoretical and experimental characterization.

It is assumed that genetic diseases affecting the metabolism of cysteine and the kidney function lead to two different kinds of pathologies, namely cystinuria and cystinosis whereby generate L-cystine crystals. Recently, the presence of L-cysteine crystal has been underlined in the case of cystinosis. Interestingly, it can be strikingly seen that cystine ([-S-CH2-CH-(NH2)-COOH]2) consists of two cysteine (C3H7NO2S) molecules connected by a disulfide (S-S) bond. Therefore, the study of cystine and cysteine is important for providing a better understanding of cystinuria and cystinosis. In this paper, we elucidate the discrepancy between L-cystine and L-cysteine by investigating the theoretical and experimental infrared spectra (IR), X-ray diffraction (XRD) as well as Raman spectra aiming to obtain a better characterization of abnormal deposits related to these two genetic pathologies.

Study on the biological behaviors of CaP coatings with different morphology on carbon/carbon composites.

In this work, we designed and fabricated a CaP composite bio-coating with different surface morphologies on a carbon/carbon (C/C) matrix by means of hybrid supersonic atmospheric plasma spraying (SAPS) and microwave-hydrothermal (MH) technologies. We found that all studied coating materials can support mesenchymal stem cells (MSCs) proliferation with prolonged culture time (3 days and 7 days) in vitro. Furthermore, according to the (Confocal Laser Scanning Microscopy) CLSM results, the MSCs also showed good attachment and different spreading morphologies on SAPS/MH coatings. As such, C/C matrix, the MH treated coatings with needle-like and rod-like microstructures were chosen for further in vivo investigation. Considering the good bonding between host tissue and the studied materials, the in vivo morphology studies confirmed a good histocompatibility for all coating samples, as well as a decreasing expression for inflammatory factors in a physiological environment. The histological results around the implants indicated different cell aggregation and vascularization ability in the local micro-environment. In particular, based on the reduction of the C/C initial surface flaws (e.g. hydrophobicity, biological inertia and easily producing carbon fragments or particles), the MH treated coating with rod-like surface morphology with a specific surface area (~2.33 m2/g) and roughness (~13.80 µm), showed excellent performance as a promising implant in live tissue.

A Root Tip-Specific Expressing Anthocyanin Marker for Direct Identification of Transgenic Tissues by the Naked Eye in Symbiotic Studies.

The Agrobacterium rhizogenes hairy root transformation system is widely used in symbiotic studies of model legumes. It typically relies on fluorescent reporters, such as DsRed, for identification of transgenic roots. The MtLAP1 transcription factor has been utilized as a reporter system in Medicago truncatula based on production of anthocyanin pigment. Here, we describe a version of this reporter driven by a root-cap specific promoter for direct observation of anthocyanin accumulation in root tips, which allows the identification of transgenic hairy roots by the naked eye. Results from our analysis suggest that the reporter had no significant effects on nodulation of M. truncatula. This approach, by virtue of its strong and specific expression in root cap cells, greatly reduces false positives and false negatives, and its use of an easily scored visible pigment should allow greater versatility and efficiency in root biology studies.

Three Common Symbiotic ABC Subfamily B Transporters in Medicago truncatula Are Regulated by a NIN-Independent Branch of the Symbiosis Signaling Pathway.

Several ATP-binding cassette (ABC) transporters involved in the arbuscular mycorrhizal symbiosis and nodulation have been identified. We describe three previously unreported ABC subfamily B transporters, named AMN1, AMN2, and AMN3 (ABCB for mycorrhization and nodulation), that are expressed early during infection by rhizobia and arbuscular mycorrhizal fungi. These ABCB transporters are strongly expressed in symbiotically infected tissues, including in root-hair cells with rhizobial infection threads and arbusculated cells. During nodulation, the expression of these genes is highly induced by rhizobia and purified Nod factors and is dependent on DMI3 but is not dependent on other known major regulators of infection, such as NIN, NSP1, or NSP2. During mycorrhization their expression is dependent on DMI3 and RAM1 but not on NSP1 and NSP2. Therefore, they may be commonly regulated through a distinct branch of the common symbiotic pathway. Mutants with exonic Tnt1-transposon insertions were isolated for all three genes. None of the single or double mutants showed any differences in colonization by either rhizobia or mycorrhizal fungi, but the triple amn1 amn2 amn3 mutant showed an increase in nodule number. Further studies are needed to identify potential substrates of these transporters and understand their roles in these beneficial symbioses.[Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Effect of sprayed techniques on the surface microstructure and in vitro behavior of nano-HAp coatings.

Supersonic atmospheric plasma spray (SAPS) technique is a classical method which is employed to coat the carbon/carbon (C/C) composites by nano-hydroxyapatite (HAp) powders to decrease the biologically inert, hydrophobic drawbacks of substrate surfaces. In recent years, profiting from the promoting of energy conservation and environmental protection, more emphasis was placed on industrial manufacturing to simplify the experimental steps. This paper aims to study the preparation of nano-HAp coatings by suspension plasma spray (SPS) instead of the original SAPS technique. A denser, more uniform and less defective coating is successfully fabricated on C/C substrate using SPS technique. More important, fewer surface flaws in SPS coating could be observed by scanning electron microscopy (SEM) which shows that the large drawbacks of the coating have disappeared during spraying process. Meanwhile, except for HAp, phase compositions of the SPS coating appear with slight calcium oxide (CaO, 0.8%) and tricalcium phosphate (TCP, 30.7%), and then all CaO as well as TCP phases transform into dicalcium phosphate anhydrous (DCPA, 60.6%) after microwave-hydrothermal (MH) treatment. Thermal analysis (TG/DSC) reveals that SPS coating (97.51%) has a higher thermal stability than that of the SAPS coating (82.37%). Also, in comparison, the SPS coating after MH treatment (SPS-MH coating) exhibits better thermal properties (92.76%). In addition, compared to the SAPS and SPS coatings, due to the more flaw reduction and phase transformation, SPS-MH coating shows a better biological properties according to the surface microstructure in simulation body fluid (SBF) solution and cell spreading area on coating. The highest corrosion resistance with the current density of 3.9798 × 10-7 A/cm2 and a potential of 0.0419 V is achieved for SPS-MH coating.

Micro-patterned surface construction on BCP ceramics and the regulation on inflammation-involved osteogenic differentiation.

The surface characteristics of bioceramics are critical in modulating the cellular events in osteogenesis. However, understanding on the effect of surface properties-derived inflammation on osteogenesis is unclear. In this study, dense biphase calcium phosphate (BCP) ceramics with various surface micropatterns were fabricated to investigate the effect of surface morphologies on the behaviors and functions of macrophages, and the subsequent cascade regulation on the osteogenic differentiation of preosteoblast. We found that the change of surface roughness and wettability of single convex in micropatterns with different sizes could make dramatic differences on the adherence, distribution and morphology of macrophages. Specifically, the patterned surface with smaller single convex area was more likely to drive macrophages towards M1-like phenotype, as was determined by the upregulated gene expression and molecular secretion of M1-specific markers compared with flat surface. Moreover, the macrophages/scaffolds-derived medium showed an obvious promotion on osteogenic differentiation of MC3T3-E1 cells that suggested the surface characteristics of BCP ceramics are important in terms of scaffolds-induced inflammation, which might contribute to its osteogenesis potential.

Mechanical properties of the supersonic atmospheric plasma sprayed CaP coating post-processed by a microwave-hydrothermal method.

Coating fabricated by supersonic atmospheric plasma spraying (SAPS) technique is post-treated by a microwave-hydrothermal (MH) method. In this case, after the surface defects are reduced, the mechanical properties between the sprayed CaP coating treated by the MH process and carbon/carbon (C/C) substrate are also studied. Accordingly, observation of the surface morphology and the cross-section of the coating are all performed by scanning electron microscopy. The phase composition and the chemical groups are examined by X-ray diffraction and Fourier Transform Infrared. Besides, a number of required measuring methods have been performed, which include shear strength, modulus of elasticity, hardness and friction coefficient. They are carried out using shear test, nanoindentation, and friction test in the mechanical property evaluation. It's obviously found that the MH method after the SAPS technique can enhance the mechanical properties at the coating-C/C substrate interface. The results show that the bonding strength of the coating after MH treatment, which is 10.46 MPa, increases by 33.25% compared to the bonding strength of the sprayed CaP coating (7.85 MPa). And the modulus of elasticity and hardness are also increased (33.74% and 38.13%). Additionally, the friction coefficient of the coating has been improved in the stable state. Finally, in simulated body fluid (SBF) experiment, the sprayed CaP coating after MH post-treatment also exhibits the bioactive behavior.

[A water maze for testing the motion of aquatic animal robots].

The existing mazes are mainly used to study the learning and memory of animals. However, there is still a lack of corresponding maze and method in the aspect of the observation and test of aquatic animal robots. For this purpose, the authors have developed a three-dimensional water maze combined with bilayer multi-channel which equips with stratified lines and tick marks. This device is a rectangular structure composed of one square bottom and four rectangular side walls, and the channels of every side wall are composed of one upper channel and two lower channels. The center of the upper channels is in the vertical center line of every side wall, and the two uper channels of adjacent side walls are at 90° degrees with each other, and the two lower channels of adjacent side walls are at 45° degrees with each other. There are stratified lines and tick marks on the side wall to test the spatial location and movement trajectories of aquatic animals. The carp robot was put into the water maze for the underwater experimental detection. The success rates of left and right steering at 135, 90 and 45 degrees as well as forward motion of the carp robots ( n = 10) were over 60%. This study showed that the device could be used to observe and test the motion of the carp robot.

Glycosylation at 11Asn on hemagglutinin of H5N1 influenza virus contributes to its biological characteristics.

A stem glycosylation site of hemagglutinin (HA) is important to the stability of the HA trimmer. A previous study shows that the stem 10/11 overlap glycosylation site of the H5 subtype avian influenza virus may influence the cleavage of HA, whereas the exact site and its effect on virulence remain unclear. In this study, site-directed mutagenesis was used to generate single or double mutant rSY-Δ10(10NNAT), rSY-Δ11(10NNSA), and rSY-Δ10/11(10NNAA) of the overlapping glycosylation site (10NNST) on the HA of A/Mallard/Huadong/S/2005(SY). By using Western blot analysis, we show that both rSY-Δ11 and rSY-Δ10/11 mutant viruses had significant delay on HA cleavage and a reduced HA molecular mass compared to the wild-type virus rSY, while the rSY-Δ10 mutant virus exhibited a similar HA molecular mass to that of the wild-type virus rSY. Interestingly, both rSY-Δ11 and rSY-Δ10/11 mutant viruses reverted their glycosylation sites at 11N after passage, indicating that 11N is a true and critical glycosylation site. Compared to the wild-type virus rSY, rSY-Δ11 and rSY-Δ10/11 mutant viruses had decreased growth rates, reduced thermo- and pH-stability, decreased pathogenicity, and limited systemic spread. Therefore, our study suggests that the 11N glycosylation site plays a key role in HA cleavage, structural stability and pathogenicity in H5 subtype avian influenza virus.

Contribution of the csgA and bcsA genes to Salmonella enterica serovar Pullorum biofilm formation and virulence.

Salmonella biofilm formation is important to environmental stress resistance and virulence. However, the roles of the csgA and bcsA genes, which affect curli protein and cellulose production, respectively, in Salmonella enterica serovar Pullorum, are unknown. Here we constructed deletions in the csgA and bcsA genes in S. enterica serovar Pullorum strain S6702 and evaluated several aspects of biofilm formation and virulence. ΔcsgA showed decreased production of curli fimbriae, while ΔbcsA had reduced cellulose production. Both mutants had a reduced ability to form biofilms. ΔcsgA was reduced in adhesion and invasion to HeLa cells and exhibited decreased intracellular proliferation in HD11 macrophages. ΔbcsA exhibited increased proliferation in HD11 cells and replicated better in chicken spleens, as compared to the wild-type strain. ΔcsgA virulence was attenuated in assays involving oral challenge of one-day-old chickens.

miR-495 sensitizes MDR cancer cells to the combination of doxorubicin and taxol by inhibiting MDR1 expression.

MDR1 is highly expressed in MDR A2780DX5 ovarian cancer cells, MDR SGC7901R gastric cancer cells and recurrent tumours. It pumps cytoplasmic agents out of cells, leading to decreased drug accumulation in cells and making cancer cells susceptible to multidrug resistance. Here, we identified that miR-495 was predicted to target ABCB1, which encodes protein MDR1. To reduce the drug efflux and reverse MDR in cancer cells, we overexpressed a miR-495 mimic in SGC7901R and A2780DX cells and in transplanted MDR ovarian tumours in vivo. The results indicated that the expression of MDR1 in the above cells or tumours was suppressed and that subsequently the drug accumulation in the MDR cells was decreased, cell death was increased, and tumour growth was inhibited after treatment with taxol-doxorubicin, demonstrating increased drug sensitivity. This study suggests that pre-treatment with miR-495 before chemotherapy could improve the curative effect on MDR1-based MDR cancer.

Ca-P bioactive coating prepared by combining microwave-hydrothermal and supersonic atmospheric plasma spraying methods.

Ca-P based coatings on carbon/carbon composite (C/C) were manufactured via a combined method comprising of microwave-hydrothermal (MH) and supersonic atmospheric plasma spraying (SAPS) techniques. However, a weak mutual interaction between the coating and C/C substrate has been a critical issue for a long time. Herein, we reported a new method for shear strength enhancement without compromising the osteoconductivity and osteoproductivity. Results showed that the inner layer has a strong mechanical interlocking with C/C substrate and the failure mode of outer layer changed from the coating cohesion (within the coating) to adhesive (at the coating/substrate interface) fracture. The shear strength between Ca-P bioactive coating-C/C substrate by MH/SAPS was significantly improved as compared to that prepared by SAPS. The Ca-P bioactive coating exhibited a good bioactivity as evidenced by the formation of a uniform carbonate-apatite layer formed on coating after immersing into stimulated body fluid for a specified period of time.

Preparation and properties of in-situ growth of carbon nanotubes reinforced hydroxyapatite coating for carbon/carbon composites.

Carbon nanotubes (CNTs) possess excellent mechanical properties for their role playing in reinforcement as imparting strength to brittle hydroxyapatite (HA) bioceramic coating. However, there are few reports relating to the in-situ grown carbon nanotubes reinforced hydroxyapatite (CNTs-HA) coating. Here we demonstrate the potential application in reinforcing biomaterials by an attempt to use in-situ grown of CNTs strengthen HA coating, using a combined method composited of injection chemical vapor deposition (ICVD) and pulsed electrodeposition. The microstructure, phases and chemical compositions of CNTs-HA coatings were characterized by various advanced methods. The scanning electron microscopy (SEM) images indicated that CNTs-HA coatings avoided the inhomogeneous dispersion of CNTs inside HA coating. The result show that the interfacial shear strength between CNTs-HA coating and the C/C composite matrix reaches to 12.86±1.43MPa. Potenitodynamic polarization and electrochemical impedance spectroscopy (EIS) studies show that the content of CNTs affects the corrosion resistance of CNTs-HA coating. Cell culturing and simulated body fluid test elicit the biocompatibility with living cells and bioactivity of CNTs-HA coatings, respectively.

[Relationship between the incidence of chronic pain and academic pressure in high school students].

OBJECTIVE: To explore the incidence of four types of chronic pain, i.e. headache, abdominal pain, neck & shoulder pain (NSP) and low back pain (LBP) and examine the relationship between the incidence of chronic pain and academic pressure in high school students. METHODS: A total of 3 000 high school students were randomly surveyed with a questionnaire on related issues. And the results were analyzed with a multivariate Logistic regression model. RESULTS: Among them, 2 849 completed the questionnaire. And the overall incident rates of headache, abdominal pain, NSP, and LBP were 30.3%, 20.9%, 32.8% and 41.1% respectively. The students generally experienced a heavy burden of learning, a high level of stress and sleep deprivation closely related to four types of chronic pain. CONCLUSION: As a common condition in Chinese adolescents, chronic pain is closely correlated with academic pressure.