Insight of endophytic fungi promoting the growth and development of woody plants.

Microorganisms play an important role in plant growth and development. In particular, endophytic fungi is one of the important kinds of microorganisms and has a mutually beneficial symbiotic relationship with host plants. Endophytic fungi have many substantial benefits to host plants, especially for woody plants, such as accelerating plant growth, enhancing stress resistance, promoting nutrient absorption, resisting pathogens and etc. However, the effects of endophytic fungi on the growth and development of woody plants have not been systematically summarized. In this review, the functions of endophytic fungi for the growth and development of woody plants have been mainly reviewed, including regulating plant growth (e.g., flowering, root elongation, etc.) by producing nutrients and plant hormones, and improving plant disease, insect resistance and heavy metal resistance by producing secondary metabolites. In addition, the diversity of endophytic fungi could improve the ability of woody plants to adapt to adverse environment. The components produced by endophytic fungi have excellent potential for the growth and development of woody plants. This review has systematically discussed the potential regulation mechanism of endophytic fungi regulating the growth and development of woody plants, it would be of great significance for the development and utilization of endophytic fungi resource from woody plants for the protection of forest resources.

Genome-wide identification and expression analysis of MYB gene family in Cajanus cajan and CcMYB107 improves plant drought tolerance.

MYB transcription factor (TF) is one of the largest superfamilies that play a vital role in multiple plant biological processes. However, the MYB family has not been comprehensively identified and functionally verified in Cajanus cajan, which is the sixth most important legume crop. Here, 170 CcR2R3-MYBs were identified and divided into 43 functional subgroups. Segmental and tandem duplications and alternative splicing events were found and promoted the expansion of the CcR2R3-MYB gene family. Functional prediction results showed that CcR2R3-MYBs were mainly involved in secondary metabolism, cell fate and identity, developmental processes, and responses to abiotic stress. Cis-acting element analysis of promoters revealed that stress response elements were widespread in the above four functional branches, further suggesting CcR2R3-MYBs were extensively involved in abiotic stress response. The transcriptome data and qRT-PCR results indicated that most of the CcR2R3-MYB genes responded to various stresses, of which the expression of CcMYB107 was significantly induced by drought stress. Overexpression of CcMYB107 enhanced antioxidant enzyme activity and increased proline and lignin accumulation, thus improving the drought resistance of C. cajan. Furthermore, Overexpression of CcMYB107 up-regulated the expression of stress-related genes and lignin biosynthesis genes after drought stress. Our findings established a strong foundation for the investigation of biological function of CcR2R3-MYB TFs in C. cajan.

Three-dimensional phenotype characteristics of skeletal class III malocclusion in adult Chinese: a principal component analysis-based cluster analysis.

BACKGROUND: Skeletal class III malocclusion has a diverse and complicated aetiology involving environmental and genetic factors. It is critical to correctly classify and define this malocclusion to be diagnosed and treated on a clinically sound basis. Thus, this study aimed to provide reliable and detailed measurements in a large ethnically homogeneous sample of Chinese adults to generate an adequate phenotypic clustering model to identify and describe the skeletal variation present in skeletal class III malocclusion. MATERIALS AND METHODS: This is a retrospective cross-sectional study in which 500 pre-treatments cone-beam computed tomography (CBCT) scans of patients with skeletal class III malocclusion (250 males and 250 females) were selected following specific selection criteria. Seventy-six linear, angular, and ratios measurements were three-dimensionally analysed using InVivo 6.0.3 software. These measurements were categorised into 47 skeletal, 18 dentoalveolar, and 11 soft tissue variables. Multivariate reduction methods: principal component analyses and cluster analyses were used to present the most common phenotypic groupings of skeletal class III malocclusion in Han ethnic group of Chinese adults. RESULTS: The principal component analysis revealed eight principal components accounted for 72.9% of the overall variation of the data produced from the seventy-six variables. The first four principal components accounted for 53.37% of the total variations. They explained the most variation in data and consisted mainly of anteroposterior and vertical skeletal relationships. The cluster analysis identified four phenotypes of skeletal class III malocclusion: C1, 34%; C2, 11.4%; C3, 26.4%; and C4, 28.2%. CONCLUSION: Based on three-dimensional analyses, four skeletal class III malocclusion distinct phenotypic variations were defined in a large sample of the adult Chinese population, showing the occurrence of phenotypic variation between identified clusters in the same ethnic group. These findings might serve as a foundation for accurate diagnosis and treatment planning of each cluster and future genetic studies to determine the causative gene(s) of each cluster.

Preparation of Nanocomposites for Antibacterial Orthodontic Invisible Appliance Based on Piezoelectric Catalysis.

Compared to fixed orthodontic appliances with brackets, thermoplastic invisible orthodontic aligners offer several advantages, such as high aesthetic performance, good comfort, and convenient oral health maintenance, and are widely used in orthodontic fields. However, prolonged use of thermoplastic invisible aligners may lead to demineralization and even caries in most patients' teeth, as they enclose the tooth surface for an extended period. To address this issue, we have created PETG composites that contain piezoelectric barium titanate nanoparticles (BaTiO3NPs) to obtain antibacterial properties. First, we prepared piezoelectric composites by incorporating varying amounts of BaTiO3NPs into PETG matrix material. The composites were then characterized using techniques such as SEM, XRD, and Raman spectroscopy, which confirmed the successful synthesis of the composites. We cultivated biofilms of Streptococcus mutans (S. mutans) on the surface of the nanocomposites under both polarized and unpolarized conditions. We then activated piezoelectric charges by subjecting the nanocomposites to 10 Hz cyclic mechanical vibration. The interactions between the biofilms and materials were evaluated by measuring the biofilm biomass. The addition of piezoelectric nanoparticles had a noticeable antibacterial effect on both the unpolarized and polarized conditions. Under polarized conditions, nanocomposites demonstrated a greater antibacterial effect than under unpolarized conditions. Additionally, as the concentration of BaTiO3NPs increased, the antibacterial rate also increased, with the surface antibacterial rate reaching 67.39% (30 wt% BaTiO3NPs). These findings have the potential for application in wearable, invisible appliances to improve clinical services and reduce the need for cleaning methods.

Purification of Polysaccharides from Orostachys cartilaginous Cell Cultures by Macroporous Resin Absorption and Bioactivities of the Purified Polysaccharides.

Orostachys cartilaginous is a traditional herbal medicine and its cell cultures contain large amounts of polysaccharides. To utilize the cultured O. cartilaginous cells, this study purified the crude polysaccharides of O. cartilaginous cells by macroporous resin absorption and optimized the purification process in the experiment of orthogonal design with four factors (sample concentration and volume, and eluent concentration and volume) and three levels; the antibacterial and anti-cancer effects of the purified polysaccharides (OTP) were further examined. The results showed that polysaccharide purity reached 95 % in the optimized group, i. e., 1.6 mg/mL of sample (crude polysaccharides) concentration, 3.0 bed volume (BV) of sample volume, 65 % eluent (ethanol) concentration, and 3.0 BV of eluent volume. In the antibacterial experiment, the growth of three bacterial species, i. e., Pseudomonas aeruginosa, Staphylococcus aureus, and Bacillus subtilis was inhibited by OTP, whereas that of Escherichia coli was not affected; the largest diameter of the inhibitory zone was found on B. subtilis; the extracellular alkaline phosphatase activity and the electrical conductivity, nucleic acid, and protein levels of B. subtilis increased after OTP treatment, indicating that the inhibition of B. subtilis growth was caused by the leakage of cell contents. In the anti-cancer experiment, OTP decreased the cell viabilities of the tested human cancer cells, i. e., AGS (gastric cancer), HCT116 (colon cancer), HepG2 (liver cancer), and HeLa (cervical cancer), and the highest inhibitory effect was on HCT116. OTP promoted HCT116 apoptosis and affected the expression of apoptosis-related proteins, i. e., the expression of B-cell lymphoma-2 decreased and that of bcl-2 associated X protein, cytochrome c, caspase 9 and caspase 3 increased. The findings of the present study suggest that O. cartilaginous cell cultures have a potential application in food or drug production.

First report of Alternaria longipes causing leaf spot on tea in China.

Tea [Camellia sinensis (L.) Kuntze)] have been widely planted in Guizhou Province in recent years, and the cultivation area in the region ranks first among all the provinces or cities in China. Leaf spot disease was an important disease of tea in Kaiyang county, Guizhou Province, which mainly damaged young leaves and shoot of tea and led to a huge loss of the production of tea. The spots initially represented brown and round, and then the diameter of the spot was 4-6 mm during later period, with the color of the center in the spot changing white. Tea leaf spot disease always occurs in early spring and the region with 1300 m altitude. From 2016 to 2019, disease incidence of leaves was estimated at 84% to 92%, and the disease severity on a plant basis was determined to be 64% to 76%, depending on the field. To identify the causal agent of the foliar disease, pieces of the lesion margins were surface sterilized with 75% ethanol for 30 s, followed by 0.5% sodium hypochlorite for 5 min, rinsed with sterile water three times, plated on potato dextrose agar (PDA) and incubated in the dark at 25C for 3 to 5 d. The hyphal tips from the margins of the growing colonies were successively picked and transferred to fresh PDA plates to purify the isolates. The result indicated that the isolates on PDA represented initially round form, and white mycelium. The reverse sides of the isolates firstly displayed light yellow on PDA. Conidiophores represent dark brown, geniculate. Brown conidia, narrow ovoid, length: 22.9 ± 4.5 µm, width: 11.1 ± 1.7 µm, with 4 to 8 transverse septa and with conspicuously ornamented walls. The gene of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Berbee et al. 1999) and the Alternaria allergen 1 (Alt a1) (Hong et al. 2005) of three strains were amplified, sequenced and deposited in Genbank. Maximum parsimony phylogenetic analysis based on concatenated sequences of combined GAPDH (1-583) and Alt a 1 (588-1065) indicated that the strain AXLKY_2019_010 was identical to reference strain Alternaria longipes strain EGS 30-033, and the clade was supported by 96% bootstrap values. According to the Koch's postulate, the tea leaves were inoculated with PDA plugs with actively growing mycelia using the methods of the puncture, cut and unwound under the laboratory conditions and the natural conditions. Slight yellow spots were gradually formed after 2 d post-inoculation on the inoculated leaves, and the color of the center of the spot changed to be white. With the prolonging of inoculation time, the size of lesion represented to be slightly enlarged. PDA plugs without mycelia were used as a control, and the control group showed no symptoms. The same isolates were consistently reisolated from inoculated leaves. A. longipes can cause leaf blight of carrots in Israel (Vintal et al. 2002), leaf spot of potato in Pakistan (Shoaib et al. 2014) and leaf spot of Atractylodes macrocephala in China (Tan et al. 2012). To our knowledge, this is the first report of A. longipes causing leaf spot on tea in China and our findings will be useful for its management and for further research.

Proteome-Scale Investigation of Protein Allosteric Regulation Perturbed by Somatic Mutations in 7,000 Cancer Genomes.

The allosteric regulation triggering the protein's functional activity via conformational changes is an intrinsic function of protein under many physiological and pathological conditions, including cancer. Identification of the biological effects of specific somatic variants on allosteric proteins and the phenotypes that they alter during tumor initiation and progression is a central challenge for cancer genomes in the post-genomic era. Here, we mapped more than 47,000 somatic missense mutations observed in approximately 7,000 tumor-normal matched samples across 33 cancer types into protein allosteric sites to prioritize the mutated allosteric proteins and we tested our prediction in cancer cell lines. We found that the deleterious mutations identified in cancer genomes were more significantly enriched at protein allosteric sites than tolerated mutations, suggesting a critical role for protein allosteric variants in cancer. Next, we developed a statistical approach, namely AlloDriver, and further identified 15 potential mutated allosteric proteins during pan-cancer and individual cancer-type analyses. More importantly, we experimentally confirmed that p.Pro360Ala on PDE10A played a potential oncogenic role in mediating tumorigenesis in non-small cell lung cancer (NSCLC). In summary, these findings shed light on the role of allosteric regulation during tumorigenesis and provide a useful tool for the timely development of targeted cancer therapies.

Are the Apo Proteins Suitable for the Rational Discovery of Allosteric Drugs?

Allosteric modulators, by targeting the less-conserved allosteric sites, represent an innovative strategy in drug discovery. These modulators have a distinctive advantage over orthosteric ligands that attach to the conserved, functional orthosteric sites. However, in structure-based drug design, it remains unclear whether allosteric protein structures determined without orthosteric ligand binding are suitable for allosteric drug screening. In this study, we performed large-scale conformational samplings of six representative allosteric proteins uncomplexed ( apo) and complexed ( holo) with orthosteric ligands to explore the effect of orthosteric site binding on the conformational dynamics of allosteric sites. The results, coupled with the redocking evaluation of allosteric modulators to their apo and holo proteins using their MD trajectories, indicated that orthosteric site binding had an effect on the dynamics of the allosteric sites and allosteric modulators preferentially bound to their holo proteins. According to the analysis data, we constructed a new correlation model for quantifying the allosteric site change driven by substrate binding to the orthosteric site. These results highlight the strong demand to select holo allosteric proteins as initial inputs in structure-based allosteric drug screening when the distance between orthosteric and allosteric sites in the protein is below 5 Å, which is expected to contribute to allosteric drug discovery.

Investigation of ECD conformational transition mechanism of GLP-1R by molecular dynamics simulations and Markov state model.

As a member of the class B G protein-coupled receptors (GPCRs), the glucagon-like peptide-1 (GLP-1) can regulate the blood glucose level by binding to the glucagon-like peptide-1 receptor (GLP-1R). Since the extracellular domain (ECD) of GLP-1R is considered as one of the binding sites of GLP-1, the open and closed states of ECD play an important role in the binding process of GLP-1. To investigate the transition path of GLP-1R ECD, the crystal structures of GLP-1R in its bound and unbound states (apo-state) are chosen to perform a total of 1.6 µs of molecular dynamics simulations. The simulated results show that the ECD of GLP-1R closes in the GLP-1 bound state and opens in the GLP-1 unbound state. To determine the critical role that GLP-1 played in regulating the open and closed states of the ECD, we applied the independent gradient model (IGM) to the simulation trajectories. We found that the "hand-like" N-terminal of the GLP-1R ECD plays an important role in the GLP-1 binding. In contrast, the apo-state GLP-1R ECD opens and exposes the two ligand binding domains of GLP-1 after 200 ns of simulations. To elucidate the open and closed mechanisms of GLP-1R ECD in the apo-state and GLP-1 bound state, the Markov state model (MSM) is performed on the MD simulation trajectories. Our results provide possible transition pathways from the closed state to open state of the apo-state GLP-1R ECD. Each pathway contains several intermediate states that correspond to different local minima in deep wells. The dynamical relationships and the most possible conversion pathway between two states are detailed through the MSM analysis. Our results profile the conformation transition mechanism of the GLP-1R ECD and will help in hypoglycemic peptide design of GLP-1R.

Classical molecular dynamics and metadynamics simulations decipher the mechanism of CBP30 selectively inhibiting CBP/p300 bromodomains.

The selective modulation of individual bromodomains (BDs) by small molecules represents an important strategy for the treatment of various cancers, considering that the BD-containing proteins share common BD structures and distinct pharmacological functions. Small molecule inhibitors targeting BDs outside of the bromodomain and extraterminal domain (BET, including BRD2-4 and BRDT) family are particularly lacking. CBP30 exhibited excellent selectivity for the transcriptional coactivators CBP (CREB binding protein) and p300 bromodomains, providing a new opportunity for designing selective non-BET inhibitors. Here, we performed classical molecular dynamics (cMD) and metadynamics simulations to reveal the selective mechanism of CBP30 binding with CBP/p300 and BRD4-BD1/BD2 bromodomains. The cMD simulations combined with binding free energy calculations were performed to compare the overall features of CBP30 binding with CBP/p300 and BRD4-BD1/BD2 bromodomains. Arg1173/1137, as the unique residue for CBP/p300, was responsible for the selective binding to CBP30 via cation-π and hydrogen bond interactions. Metadynamics simulation, together with unbinding free energy profiles, suggested that the dissociation pathways of CBP30 from CBP/p300 and BRD4-BD1/BD2 bromodomains were different, with the unbinding of the former being more difficult. These findings will be helpful for novel CBP/p300-inhibitor design and rational structural modification of existing inhibitors to increase their selectivity.

Revealing the conformational dynamics of UDP-GlcNAc recognition by O-GlcNAc transferase via Markov state model.

The O-linked N-acetylglucosamine (O-GlcNAc) glycosylation is a critical post-translational modification and closely linked to various physiological and pathological conditions. The O-GlcNAc transferase (OGT) functions as the only glycosyltransferase of O-GlcNAc glycosylation by transferring GlcNAc from UDP-GlcNAc to serine or threonine residues on protein substrates. The interaction mode of UDP-GlcNAc against OGT has been preliminarily revealed by the crystal structures, yet an atomic-level comprehension for the conformational dynamics of the recognition process remains elusive. Here, we construct the Markov state model based on extensive all-atom molecular dynamics (MD) simulations with an aggregated simulation time of ∼9 µs, and reveal that the UDP-GlcNAc recognition process by OGT encompasses four key metastable states, occurring within an estimated timescale of ∼10 µs. During UDP-GlcNAc recognition process, we find the pyrophosphate moiety (P2O52-) initially anchors to the active pocket via salt bridge and hydrogen bonds, facilitating subsequent binding of the uridine and GlcNAc moieties. Furthermore, the functional roles of K842 involved in the salt bridge with P2O52- were evaluated through extra mutant MD simulations. Overall, our study provides valuable insights into the UDP-GlcNAc recognition mechanism by OGT, which could further aid in mechanistic studies of O-GlcNAc glycosylation and drug development targeting on OGT.

Effects of nitrogen, phosphorus and potassium formula fertilization on the yield and berry quality of blueberry.

Through the application ratio of nitrogen (N), phosphorus (P) and potassium (K) in the field, L9 (33) orthogonal experimental design was used to study the effects of different N, P and K ratios on the yield and quality of blueberry fruit, aiming to optimize the amount of supplied fertilizers. The results showed that N, P and K fertilizer had different effects on fruit yield and quality, among which K fertilizer was the most important factor. Fertilization could significantly improve the yield and fruit quality of blueberry, and the average yield of fertilization treatment was 37.78% higher than that of the control group (CK). Even the treatment with the worst results F6 (N2P3K1), its single fruit weight, anthocyanins, total phenols, soluble solids and soluble protein content were 1.09, 1.32, 1.23, 1.08 and 1.21 times higher than the control (CK), respectively. Based on the comprehensive evaluation of principal component analysis and multi factor analysis of variance, the best fertilization combination for high-yield and good-quality blueberries was N1P2K2 (F2), that is, the best fertilization effect was that including N 100 g/plant, P2O5 25 g/plant, K2O 25 g/plant, applied in the form of ammonium sulfate (472 g/plant), superphosphate (41 g/plant) and potassium sulfate (40 g/plant), respectively.

Physiological response of anthocyanin synthesis to different light intensities in blueberry.

Fruit color is an important economic character of blueberry, determined by the amount of anthocyanin content. Anthocyanin synthesis within the blueberry fruits is significantly affected by light. To reveal the physiological response mechanism of anthocyanin synthesis in blueberry fruits in different light intensities, four light intensities (100% (CK), 75%, 50% and 25%) were set for the 'O'Neal' southern highbush blueberry as the experimental material in our study. The relationship between endogenous hormones content, associated enzyme activities, and variations with the anthocyanin content in blueberry fruits under various light intensities during the white fruit stage (S1), purple fruit stage (S2), and blue fruit stage (S3) were studied. The results showed that adequate light could significantly promote anthocyanin synthesis in blueberry fruits (P < 0.05). Blueberry fruits had an anthocyanin content that was 1.76~24.13 times higher under 100% light intensity than it was under non-full light intensity. Different light intensities significantly affected the content of endogenous hormones and the activity of associated enzymes in anthocyanin synthesis pathway (P < 0.05). Among them, the JA (jasmonic acid) content and PAL (phenylalanine ammonia lyase) activity of fruits under 100% light intensity were 2.49%~41.83% and 2.47%~48.48% higher than those under other light intensity, respectively. And a significant correlation was found between the variations in anthocyanin content in fruits and the content or activities of JA, ABA (abscisic acid), ETH (ethylene), GA3 (gibberellin 3), IAA (indoleacetic acid), PAL, CHI (chalcone isomerase), DFR (dihydroflavonol reductase) and UFGT (UDP-glucose: flavonoid 3-glucosyltransferase) (P < 0.05). It indicated that 100% light intensity significantly promoted anthocyanin synthesis in blueberry fruits by affecting endogenous hormones content and associated enzyme activities in the anthocyanin synthesis pathway. This study will lay a foundation for further research on the molecular mechanism of light intensity regulating anthocyanin synthesis in blueberry.

Impact of molar teeth distalization by clear aligners on temporomandibular joint: a three-dimensional study.

BACKGROUND: Maxillary molar distalization is a common technique used in the non-extraction treatment of Angle Class II malocclusion that can effectively correct the molar relationship and create spaces for anterior teeth alignment. However, this approach may also impact the temporomandibular joint (TMJ) due to predictable changes in the posterior vertical dimension. Despite its widespread use, Class II malocclusions correction by molar distalization with clear aligners has not been investigated for their effects on the TMJ. Therefore, this study aimed to analyze the impact of sequential molar distalization using clear aligners on the TMJ. METHODS: Three-dimensional CBCT scans of 23 non-growing patients (7 males, 16 females; mean age 29.8 ± 4.6 years) with skeletal class I or II malocclusion and a bilateral molar class II relationship treated by sequential upper molars distalization with orthodontic clear aligners (Invisalign, Align Technology, San Josè, Ca, USA). A total of 46 joints were examined before and after molar distalization using Anatomage InvivoDental 6.0.3. Linear and angular measurements of the mandibular joint were measured, including joint parameters, inclination, position, and the dimension of the condyle and articular fossa. In addition, 3D volumetric spaces of the joint were analyzed. All data were statistically analyzed by paired T test to determine the differences between the pre-and post-orthodontic procedures. RESULTS: No statistically significant differences were found in all primary effects resulting from maxillary molars distalization by clear aligners on TMJ components measurements and joint spaces between T0 and T1. Meanwhile, statistically significant differences were observed in the linear position of the upper molars and the molar relationship parameter with at least P ≤ 0.05. CONCLUSION: Treatment by sequential upper molars distalization with clear aligners does not lead to significant TMJ parameters changes in condyle and fossa spaces, dimensions, and positions.

Correlation between the three-dimensional maxillomandibular complex parameters and pharyngeal airway dimensions in different sagittal and vertical malocclusions.

OBJECTIVES: This study aimed to determine the three-dimensional (3D) correlation between maxillomandibular complex parameters and pharyngeal airway dimensions in different sagittal and vertical malocclusions. METHODS: This retrospective cross-sectional study included the CBCT scans of 368 patients with a mean age of 23.81 ± 3.01 years. The patients were classified into three groups (skeletal Class I, II, and III). Each class group was divided into three subgroups based on vertical growth patterns (hypo-, normo-, and hyperdivergent). The maxillomandibular complex was evaluated in the three planes using 16 skeletal measurements. Naso-, oro-, hypo-, and total pharyngeal airway spaces were assessed in terms of width, volume, surface area, and minimum constricted area (MCA). Two-way ANOVA followed by the Bonferroni post-hoc test were used. RESULTS: The nasopharyngeal airway space was significantly lowest regarding sagittal and lateral widths in the skeletal Class III patients, the lowest volume and surface area were in hyperdivergent patients, and MCA was the highest in Class II and hypodivergent patients. The oro- and hypopharyngeal sagittal width, volume, surface area, and MCA were the lowest in the hyperdivergent patients, and oropharyngeal lateral width and hypopharyngeal sagittal width were the highest in skeletal Class III. The total pharyngeal volume, surface area, and MCA were the lowest in the hyperdivergent patients, and skeletal Class II patients had the lowest MCA. CONCLUSIONS: The pharyngeal airway dimensions differ with various sagittal and vertical malocclusions. These differences could apply to diagnosis, treatment planning, and possible changes following orthodontic/orthopedic or surgical treatment.

Loss of RPS27a expression regulates the cell cycle, apoptosis, and proliferation via the RPL11-MDM2-p53 pathway in lung adenocarcinoma cells.

BACKGROUND: Depletion of certain ribosomal proteins induces p53 activation, which is mediated mainly by ribosomal protein L5 (RPL5) and/or ribosomal protein L11 (RPL11). Therefore, RPL5 and RPL11 may link RPs and p53 activation. Thus, this study aimed to explore whether RPs interact with RPL11 and regulate p53 activation in lung adenocarcinoma (LUAD) cells. METHODS: The endogenous RPL11-binding proteins in A549 cells were pulled down through immunoprecipitation and identified with a proteomics approach. Docking analysis and GST-fusion protein assays were used to analyze the interaction of ribosomal protein S27a (RPS27a) and RPL11. Co-immunoprecipitation and in vitro ubiquitination assays were used to detect the effects of knockdown of RPS27a on the interaction between RPS27a and RPL11, and on p53 accumulation. Cell cycle, apoptosis, cell invasion and migration, cell viability and colony-formation assays were performed in the presence of knockdown of RPS27a. The RPS27a mRNA expression in LUAD was analyzed on the basis of the TCGA dataset, and RPS27a expression was detected through immunohistochemistry in LUAD samples. Finally, RPS27a and p53 expression was analyzed through immunohistochemistry in A549 cell xenografts with knockdown of RPS27a. RESULTS: RPS27a was identified as a novel RPL11 binding protein. GST pull-down assays revealed that RPS27a directly bound RPL11. Knockdown of RPS27a weakened the interaction between RPS27a and RPL11, but enhanced the binding of RPL11 and murine double minute 2 (MDM2), thereby inhibiting the ubiquitination and degradation of p53 by MDM2. Knockdown of RPS27a stabilized p53 in an RPL11-dependent manner and induced cell viability inhibition, cell cycle arrest and apoptosis in a p53-dependent manner in A549 cells. The expression of RPS27a was upregulated in LUAD and correlated with LUAD progression and poorer prognosis. Overexpression of RPS27a correlated with upregulation of p53, MDM2 and RPL11 in LUAD clinical specimens. Knockdown of RPS27a increased p53 activation, thus, suppressing the formation of A549 cell xenografts in nude mice. CONCLUSIONS: RPS27a interacts with RPL11, and RPS27a knockdown enhanced the binding of RPL11 and MDM2, thereby inhibiting MDM2-mediated p53 ubiquitination and degradation; in addition, RPS27a as important roles in LUAD progression and prognosis, and may be a therapeutic target for patients with LUAD.

3D Finite Element Study of the Physiological Anchorage Control Concept on Anchorage Molars in Lingual Orthodontics.

Objective: To study the effect of the physiological anchorage control concept on anchorage molars in lingual and labial orthodontic techniques. Methods: Three-dimensional finite element models, including the right maxillary first molar, periodontal ligament, alveolar bone, and buccal tube, were established. The models were divided into the McLaughlin-Bennett-Trevisi (MBT™) straight-wire model with 0-degree maxillary first molar axial inclination and the physiologic anchorage Speewire system (PASS) model with -7-degree maxillary first molar axial inclination. Simulated sliding retraction forces (1 N, 1.5 N, and 2 N) were loaded on the buccal side and lingual side, and retraction forces (0.5 N, 0.75 N, and 1 N) were loaded on the buccal and lingual sides simultaneously. The displacements, principal stresses, and von Mises stresses of the periodontal ligament under different conditions were derived. Results: The anchorage molars showed different degrees of rotation, tipping, intrusion, and extrusion. As the force increased, these displacement trends also increased. The mesial displacement of the buccal + lingual force loading was less than that of the other two groups. Under the same force load method, the mesial displacement of the PASS group was less than that of the MBT group. Tilt movement increases the tensile stress of the distal cervical margin and root mesial apical third and the compressive stress of the mesial cervical margin and root distal apical third. The maximum stress of the periodontal ligament was less than that of the other two groups when the lingual force was loaded. Conclusion: The physiological anchorage control concept in lingual orthodontics provides better sagittal anchorage control than in labial orthodontics, but there is no significant difference numerically. Attention should be given to the control of torsion, torque, and arch width. Tilt movement increases the PDL stress of the cervical margin and root apical third. The sliding retraction force should be loaded lingually to maintain the force value of 1∼1.5 N.

The preparation of lactoferrin/magnesium silicate lithium injectable hydrogel and application in promoting wound healing.

The development of novel wound dressings with highly effective antibacterial and accelerating wound healing properties has become the focus of current research. In this study, a novel and injectable lactoferrin (LF)/lithium magnesium silicate hydrogel (LMSH) was first synthesized through a simple electrostatic interaction method. The physical and biological properties are systematically characterized. The results show that the synthesized LF/LMSH has good antibacterial properties and biocompatibility. More importantly, it can effectively promote wound healing in the rat full-thickness skin wound model after 14 days post-operation, and the healing rate can reach 99.1 %, which is much higher than that of other groups. Meanwhile, histochemical and immunofluorescent staining confirm that the prepared injectable LF/LMSH has good pro-collagen deposition, pro-angiogenic and anti-inflammatory properties. The healed wounds present a consistently thickened epidermis with more follicular and glandular structures, indicating the great potential of the prepared material for wound management.

Recurrent neural network (RNN) model accelerates the development of antibacterial metronidazole derivatives.

Metronidazole is a specific drug against trichomonas and anaerobic bacteria, and is widely used in the clinic. However, extensive clinical application is often accompanied by extensive side effects, so it is still of great significance to develop metronidazole derivatives with a new skeleton. Compared with other traditional receptor-based drug design methods, the computational model based on a neural network has higher accuracy and reliability. In this work, a Recurrent Neural Network (RNN) model is applied to the discovery of metronidazole drugs with a new skeleton. Firstly, the generation model based on a Gated Recurrent Unit (GRU) is trained to generate an effective Simplified Molecular-Input Line-Entry System (SMILES) string library with high precision. Then, transfer learning is introduced to fine-tune the GRU model, and many molecules with structures similar to known active drugs are generated. After cluster analysis of the structures of the new compounds, 20 small molecular compounds with metronidazole structures of all different categories were selected, of which 19 may not belong to any published patents or applications. Through prediction and personal experience, the difficulty of synthesizing these 20 new structures was analyzed, and compound 0001 was chosen as our synthetic target, and a series of structures (8a-l) similar to compound 0001 were synthesized. Finally, the inhibitory activities of these compounds against bacteria E. coli, P. aeruginosa, B. subtilis and S. aureus were determined. The results showed that compound 8a-l had obvious inhibitory activity against these four bacteria, which proved the accuracy of our compound generation model.

Aramid nanofibers reinforced polyvinyl alcohol/tannic acid hydrogel with improved mechanical and antibacterial properties for potential application as wound dressing.

The poor mechanical properties and the lack of antibacterial ability of hydrogels limit their applications as wound dressing. In this work, a novel and high strength polyvinyl alcohol (PVA)/tannic acid (TA) hydrogel with aramid nanofibers (ANFs) as the reinforcement was successfully fabricated. The surface composition and microstructure of the hydrogel were characterized by fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and transmission electron microscopy (TEM). The mechanical properties, water content and swelling behaviors, as well as the antibacterial abilities and biocompatibility of the prepared hydrogel were systematically analyzed as well. The results indicated that the prepared hydrogel showed excellent mechanical properties. The tensile strength and elongation of the prepared hydrogel can respectively reach 2.06 MPa and 950% owing to the formation of the multiple H bonds among PVA, ANFs and TA. What's more, PVA/ANFs/TA (PAT) hydrogel possessed shape memory and broad-spectrum antibacterial properties against S. aureus, E. coli and P. aeruginosa (100% antibacterial rate) at the concentration of 12 mg/mL. PAT hydrogels also had low cytotoxicity, affirming its potential application as wound dressing.