Comparative Study of the Efficacy of Autologous Tooth Bone Powder and Inorganic Bovine Bone Powder on the Repair of Jaw Defects.

Context: The main pathological features of jaw cysts are bone defects. Obtaining autologous bone for transplantation repair has been associated with postoperative complications, and the amount of bone that dentist can collect is limited. Studies have found that autologous tooth bone powder is safe and has good bone-formation ability and stability. Objective: The study intended to examine the efficacy of implantation of autologous tooth bone powder and inorganic bovine bone powder, after marsupialization and second-stage curettage for large jaw cysts that dentist can't directly remove by surgery in clinical practice. Design: The research team designed a prospective randomized controlled trial. Setting: The study took place in the Head and Neck Surgery Department at Chongqing University Cancer Hospital in Chongqing, China. Participants: Participants were 60 patients at the hospital between 2016 and 2018 who had mandibular cysts that surgical operation couldn't directly remove by surgery in clinical practice. Intervention: At 4 months after curettage, the research team randomly divided participants into three groups: (1) an intervention group who received implants of autologous tooth bone powder into the bone defects, (2) a positive control group who received implants of inorganic bovine bone powder, and (3) a negative control group who received no implants of any material. Outcome Measures: The research team performed: (1) periodontal probing at a fixed anatomical point for the intervention and both control groups postintervention at one day and 4 months after surgery and recorded the changes in probing depth and (2) computed tomography (CT) scans at baseline one day before and postintervention at 4 months after the implantation to determine changes in the bone mineral density and compared them among the three groups. Results: The change in the height of the intervention group's fixed anatomical point postintervention at 4 months after surgery was significantly smaller than that of the positive control group (P < .05). In the CT scan analysis, the differences between the intervention and negative control groups and between the positive and negative control groups were statistically significant (P < .05); however, the difference between the intervention and positive control groups wasn't significant (P > .05). Conclusions: Autologous tooth bone powder and inorganic bovine bone powder can effectively repair bone defects caused by large jaw cysts and that the repaired effect may be better than that of spontaneous osteogenesis. The autologous tooth bone powder was associated with lower levels of bone loss than those seen with use of inorganic bovine bone powder.

Real-Time Dissecting the Dynamics of Drug Transportation in the Live Brain.

Brain diseases are becoming a more and more serious threat to human health. Many critical properties of the transport mechanisms of drugs in live brains remain poorly understood. In this work, single-particle tracking was used to dissect the transport dynamics of wheat germ agglutinin (WGA) in live brain and characterize the geometry and rheology of the extracellular space (ECS). The results revealed that the movements of WGA were influenced by the specific-binding molecules and the nature of the ECS. We further analyzed the mobility behaviors of WGA globally and quantitatively and found that movement of WGA in brain cells of acute slices was an active transport process associated with actin filaments and microtubules. This work paves the way for studies aiming at characterizing the biophysics of drug transport in the context of live brains, which may contribute to developing potential new therapeutic applications for brain diseases.

Food-grade γ-aminobutyric acid production by immobilized glutamate decarboxylase from Lactobacillus plantarum in rice vinegar and monosodium glutamate system.

OBJECTIVES: γ-amino butyric acid (GABA) is a non-protein amino acid, considered a potent bioactive compound. This study focused on biosynthesis of food-grade GABA by immobilized glutamate decarboxylase (GAD) from Lactobacillus plantarum in the rice vinegar and monosodium glutamate (MSG) reaction system. RESULTS: The gene encoding glutamate decarboxylase (GadB) from L. plantarum has been heterologously expressed in Lactococcus lactis and biochemically characterized. Recombinant GadB existed as a homodimer, and displayed maximal activity at 40 °C and pH 5.0. The Km value and catalytic efficiency (kcat/Km) of GadB for L-Glu was 22.33 mM and 62.4 mM-1 min-1, respectively, with a specific activity of 24.97 U/mg protein. Then, purified GadB was encapsulated in gellan gum beads. Compared to the free enzyme, immobilized GadB showed higher operational and storage stability. Finally, 9.82 to 21.48 g/L of GABA have been acquired by regulating the amounts of catalyst microspheres ranging from 0.5 to 0.8 g (wet weight) in 0.8 mL of the designed rice vinegar and MSG reaction system. CONCLUSIONS: The method of production GABA by immobilized GadB microspheres mixed in the rice vinegar and MSG reaction system is introduced herein for the first time. Especially, the results obtained here meet the increased interest in the harnessing of biocatalyst to synthesize food-grade GABA.

A Single Mutation Increases the Thermostability and Activity of Aspergillus terreus Amine Transaminase.

Enhancing the thermostability of (R)-selective amine transaminases (AT-ATA) will expand its application in the asymmetric synthesis of chiral amines. In this study, mutual information and coevolution networks of ATAs were analyzed by the Mutual Information Server to Infer Coevolution (MISTIC). Subsequently, the amino acids most likely to influence the stability and function of the protein were investigated by alanine scanning and saturation mutagenesis. Four stabilized mutants (L118T, L118A, L118I, and L118V) were successfully obtained. The best mutant, L118T, exhibited an improved thermal stability with a 3.7-fold enhancement in its half-life (t1/2) at 40 °C and a 5.3 °C increase in T5010 compared to the values for the wild-type protein. By the differential scanning fluorimetry (DSF) analysis, the best mutant, L118T, showed a melting temperature (Tm) of 46.4 °C, which corresponded to a 5.0 °C increase relative to the wild-type AT-ATA (41.4 °C). Furthermore, the most stable mutant L118T displayed the highest catalytic efficiency among the four stabilized mutants.

The pretreatment neutrophil-lymphocyte ratio may predict prognosis of patients with liver cancer: A systematic review and meta-analysis.

BACKGROUND: At present, several studies have reported that the pretreatment neutrophil-lymphocyte ratio (NLR) may be associated with the prognosis of liver cancer. Nevertheless, their conclusions remain controversial. Thus, we performed a meta-analysis of 54 studies to evaluate the prognostic value of NLR. METHOD: Databases including PubMed, Embase, Cochrane Library, and Web of Science were searched to July 2017. RESULT: A total of 54 studies including 12 979 patients were included in this meta-analysis. Elevated NLR had a close relationship with the overall survival (OS) (HR 1.52; 95% CI 1.39-1.67), recurrence-free survival (RFS) (HR 1.84; 95% CI 1.48-2.30), and disease-free survival (DFS) (HR 1.71; 95% CI 1.39-2.11) of liver cancer, respectively. In addition, elevated NLR was associated with the presence of tumor vascular invasion (OR 2.35; 95% CI 1.93-2.86), multiple tumors (OR 1.38; 95% CI 1.15-1.66), alpha-fetoprotein ≥ 400 ng/mL (OR 1.51; 95% CI 1.15-1.98), presence of HbsAg (+) (OR 0.68; 95% CI 0.51-0.90), and cirrhosis (OR: 0.59; 95% CI 0.44-0.80). CONCLUSION: This meta-analysis indicated that elevated NLR may be an effective and noninvasive indicator for prognosis of patients with liver cancer.

Improving thermostability of (R)-selective amine transaminase from Aspergillus terreus through introduction of disulfide bonds.

To improve the thermostability of (R)-selective amine transaminase from Aspergillus terreus (AT-ATA), we used computer software Disulfide by Design and Modelling of Disulfide Bonds in Proteins to identify mutation sites where the disulfide bonds were most likely to form. We obtained three stabilized mutants (N25C-A28C, R131C-D134C, M150C-M280C) from seven candidates by site-directed mutagenesis. Compared to the wild type, the best two mutants N25C-A28C and M150C-M280C showed improved thermal stability with a 3.1- and 3.6-fold increase in half-life (t1/2 ) at 40 °C and a 4.6 and 5.1 °C increase in T5010 . In addition, the combination of mutant R131C-D134C and M150C-M280C displayed the largest shift in thermostability with a 4.6-fold increase in t1/2 at 40 °C and a 5.5 °C increase in T5010 . Molecular dynamics simulation indicated that mutations of N25C-A28C and M150C-M280C lowered the overall root mean square deviation for the overall residues at elevated temperature and consequently increased the protein rigidity. The stabilized mutation of R131C-D134C was in the region of high mobility and on the protein surface, and the disulfide bond constraints the flexibility of loop 121-136.

Adsorption of Cu(II) and Zn(II) Ions from Aqueous Solution by Gel/PVA-Modified Super-Paramagnetic Iron Oxide Nanoparticles.

Super-paramagnetic iron oxide nanoparticles (SPIONs)/gelatin (gel)/polyvinyl alcohol (PVA) nanoparticles were designed and synthesized by the co-precipitation method and further modified with gel and PVA. These nanoparticles were used for the removal of Cu(II) and Zn(II) from aqueous solutions. The adsorbents were rich in different functional groups for chemisorption and showed effective adsorption properties. The adsorption of Cu(II) and Zn(II) on the SPIONs/gel and SPIONs/gel/PVA materials were investigated with respect to pH, adsorption kinetics, and adsorption isotherms. The adsorption data was fitted to the Langmuir, Freundlich, and Sips models at the optimum pH 5.2 (±0.2) over 60 min; SPIONs/gel showed maximum adsorption capacities of 47.594 mg/g and 40.559 mg/g for Cu(II) and Zn(II); SPIONs/gel/PVA showed those of 56.051 mg/g and 40.865 mg/g, respectively. The experimental data fitted the pseudo-second-order model, indicating that the process followed chemical monolayer adsorption. In addition, the SPIONs/gel/PVA showed better stability and Cu(II) adsorption efficiency than SPIONs/gel.

Vesicle formation by proton transfer driven short-tailed fatty acids of C4-C8 chain length in water.

Ultrashort single-chain fatty acids self-assemble to form vesicles under certain proton-driven conditions. The protonation provides a larger charge area around the hydrophilic carbonyl headgroups, and proton shift as the key driving parameter was studied. The ultrashort fatty acids (C4-C8) formed stable unilamellar vesicles predominantly through out the whole range of tested pH levels (6.5-9.5). A proton-driven self-assembly process and effects on the phase transition were characterized by dynamic light scattering, transmission electron microscopy and cryo-transmission electron microscopy. In particular, we studied in greater detail the molecular packing characteristics of FA vesicles for geometric reasons and the protonation effect changes the molecular surface charge and further carboxylic acid headgroup motion. This study enhances the understanding of the physicochemical specificity of these membrane vesicles, and may facilitate the alteration of membrane function caused by FAs.

Fusaroside, a unique glycolipid from Fusarium sp., an endophytic fungus isolated from Melia azedarach.

Fusaroside (1), a unique trehalose-containing glycolipid composed of the 4-hydroxyl group of a trehalose unit attached to the carboxylic carbon of a long-chain fatty acid, was isolated from the organic extract of fermentation broths of an endophytic fungus, Fusarium sp. LN-11 isolated from the leaves of Melia azedarach. Six known compounds, phalluside (2), (9R*,10R*,7E)-6, 9,10-trihydroxyoctadec-7-enoic acid (3), porrigenic acid (4), (9Z)-2,3-dihydroxypropyl octadeca-9-enoate (5), cerevisterol (6) and ergokonin B (7), were also isolated from this fungus. The glycolipid contains a rare branched long-chain fatty acid (C(20:4)) with a conjugated diene moiety and a conjugated ketone moiety. The structure of the new compound 1 was elucidated by spectroscopic methods (1D and 2D NMR experiments, MS) and chemical degradations. The metabolites 1-5 were shown to have moderate to weak active against the brine shrimp larvae. To our knowledge, this is the first report of isolation of the first representative of a new family of glycolipids from natural sources.

Wavelet-Based Texture Reformation Network for Image Super-Resolution.

Most reference-based image super-resolution (RefSR) methods directly leverage the raw features extracted from a pretrained VGG encoder to transfer the matched texture information from a reference image to a low-resolution image. We argue that simply operating on these raw features neglects the influence of irrelevant and redundant information and the importance of abundant high-frequency representations, leading to undesirable texture matching and transfer results. Taking the advantages of wavelet transformation, which represents the contextual and textural information of features at different scales, we propose a Wavelet-based Texture Reformation Network (WTRN) for RefSR. We first decompose the extracted texture features into low-frequency and high-frequency sub-bands and conduct feature matching on the low-frequency component. Based on the correlation map obtained from the feature matching process, we then separately swap and transfer wavelet-domain features at different stages of the network. Furthermore, a wavelet-based texture adversarial loss is proposed to make the network generate more visually plausible textures. Experiments on four benchmark datasets demonstrate that our proposed method outperforms previous RefSR methods both quantitatively and qualitatively. The source code is available at https://github.com/zskuang58/WTRN-TIP.

Determination of Vancomycin and Norvancomycin Residues in Milk by Automated Online Solid-Phase Extraction Combined With Liquid Chromatography-High Resolution Mass Spectrometry.

BACKGROUND: Vancomycin and norvancomycin, as potent antibacterial retention drugs, were used illegally on animals bred for food, which directly affected the quality and safety of animal-derived food, and even harmed human health. OBJECTIVE: A fast analysis method, which was adopted to detect residues of vancomycin and norvancomycin in milk, was implemented on a chromatographic system containing online solid-phase extraction (SPE) device that combined with high-resolution mass spectrometer (HRMS). METHOD: First, the analytes were added to the blank milk sample were extracted with water [containing 0.1% trifluoroacetic acid (TFA)]-acetonitrile (ACN) (8:2, v/v), and then were purified and enriched on a C18-XL column, whereafter eluted from the purification column onto the analytical column (Shiseido Capcell Pak ADME column) for chromatographic separation prior to hybrid quadrupole-Orbitrap (Q-Orbitrap) detection. RESULTS: The results showed that the limit of detection (LOD) for each analyte and the limit of quantitation (LOQ) were 0.15 and 0.5 µg/kg, respectively. The correlation coefficient(s) of vancomycin and norvancomycin ranged from 0 to 200 ng/mL were greater than 0.9983. CONCLUSIONS: These validations reflected that it was suitable for the established method to rapidly analyze vancomycin and norvancomycin residues in milk. HIGHLIGHTS: The method for detecting vancomycin and norvancomycin residues in milk by online SPE combined with LC-HRMS. Online SPE technology realized automation, and the application of HRMS greatly improved the reliability of qualitative and quantitative analyses. The developed method is fast, simple, and reliable; each methodological index can meet requirements of trace analyses of vancomycin and norvancomycin in milk.

Proximity-induced exponential amplification reaction triggered by proteins and small molecules.

We proposed a method to regulate nucleic acid polymerization by proximity and designed an ultrasensitive biosensor based on proximity-induced exponential amplification reaction for proximity assay of proteins (streptavidin) and small molecules (adenosine triphosphate), which allows us to detect a variety of interesting targets by simply changing the binding sites of DNA.

Improving the Thermostability and Activity of Transaminase From Aspergillus terreus by Charge-Charge Interaction.

Transaminases that promote the amination of ketones into amines are an emerging class of biocatalysts for preparing a series of drugs and their intermediates. One of the main limitations of (R)-selective amine transaminase from Aspergillus terreus (At-ATA) is its weak thermostability, with a half-life (t 1/2) of only 6.9 min at 40°C. To improve its thermostability, four important residue sites (E133, D224, E253, and E262) located on the surface of At-ATA were identified using the enzyme thermal stability system (ETSS). Subsequently, 13 mutants (E133A, E133H, E133K, E133R, E133Q, D224A, D224H, D224K, D224R, E253A, E253H, E253K, and E262A) were constructed by site-directed mutagenesis according to the principle of turning the residues into opposite charged ones. Among them, three substitutions, E133Q, D224K, and E253A, displayed higher thermal stability than the wild-type enzyme. Molecular dynamics simulations indicated that these three mutations limited the random vibration amplitude in the two α-helix regions of 130-135 and 148-158, thereby increasing the rigidity of the protein. Compared to the wild-type, the best mutant, D224K, showed improved thermostability with a 4.23-fold increase in t 1/2 at 40°C, and 6.08°C increase in T 50 10 . Exploring the three-dimensional structure of D224K at the atomic level, three strong hydrogen bonds were added to form a special "claw structure" of the α-helix 8, and the residues located at 151-156 also stabilized the α-helix 9 by interacting with each other alternately.

[Effect of decompression combined with curettage and autogenous bone cement implantation on large cysts of the jaw].

Decompression and curettage can result are effective as treatments for large jaw cysts, which are common diseases in the clinic. Based on a treatment used in a previous study, this paper proposes a "three-step method" to treat large jaw cyst and repair the bone defect by decompression, curettage, and autologous dental bone powder implantation. This paper introduces the processes and key points of the operation involved in the abovementioned method.

Overexpression of mitogen-activated protein kinase kinase 4 and nuclear factor-kappaB in laryngeal squamous cell carcinoma: a potential indicator for poor prognosis.

This study aimed to investigate the expression and clinical significance of mitogen-activated protein kinase kinase 4 MKK4 and nuclear factor-kappaB (NF-kappaB) in patients with laryngeal squamous cell carcinoma (LSCC). We used immunohistochemistry (IHC) to examine the expression of MKK4 and NF-kappaB in 78 LSCCs and their adjacent normal tissues. To clarify the validity of MKK4 and NF-kappaB as determined by the IHC analysis, RT-PCR was performed on 21 tissues randomly selected from the 78 LSCCs. The positive expression rates of MKK4 and NF-kappaB in patients with LSCC were 67.9% (53/78) and 60.3% (47/78) respectively, which were significantly higher than those in the adjacent normal tissue (both P<0.01). The positive expression of MKK4 and NF-kappaB tended to be associated positively with lymph node metastasis (both P<0.01) as well as T stage (both P<0.01). The Spearman analysis indicated that the expression level of MKK4 was positively correlated with that of NF-kappaB significantly (rs=0.368, P<0.01). Overall survival curves estimated by Kaplan-Meier showed that tumor patients with low MKK4 and NF-kappaB expression in their tumor cells survive significantly longer than patients with high MKK4 and NF-kappaB levels (P=0.027, and P<0.01, respectively). In addition, multivariate Cox regression analysis showed that N stage, T stage and NF-kappaB expression are significant independent prognostic factors for overall survival (P<0.01, P=0.014, and P=0.027, respectively). These findings suggested that the expression of MKK4 and NF-kappaB may be considered as a useful prognostic marker of LSCC after surgical resection.

Construction of stabilized (R)-selective amine transaminase from Aspergillus terreus by consensus mutagenesis.

Amine transaminases are a class of efficient and industrially-desired biocatalysts for the production of chiral amines. In this study, stabilized variants of the (R)-selective amine transaminase from Aspergillus terreus (AT-ATA) were constructed by consensus mutagenesis. Using Consensus Finder (http://cbs-kazlab.oit.umn.edu/), six positions with the most prevalent amino acid (over 60% threshold) among the homologous family members were identified. Subsequently, these six residues were individually mutated to match the consensus sequence (I77 L, Q97E, H210N, N245D, G292D, and I295 V) using site-directed mutagenesis. Compared to that of the wild-type, the thermostability of all six single variants was improved. The H210N variant displayed the largest shift in thermostability, with a 3.3-fold increase in half-life (t1/2) at 40 °C, and a 4.6 °C increase in T5010 among the single variants. In addition, the double mutant H210N/I77L displayed an even larger shift with 6.1-fold improvement of t1/2 at 40 °C, and a 6.6 °C increase in T5010. Furtherly, the H210N/I77L mutation was introduced into the previously engineered thermostable AT-ATA by the introduction of disulfide bonds, employing B-factor and folding free energy (ΔΔGfold) calculations. Our results showed that the combined variant H210N/I77L/M150C-M280C had the largest shift in thermostability, with a 16.6-fold improvement of t1/2 and a 11.8 °C higher T5010.

LTBP2 promotes the migration and invasion of gastric cancer cells and predicts poor outcome of patients with gastric cancer.

Latent transforming growth factor-ß-binding protein (LTBP)2 is a member of the fibrillin/LTBP superfamily of extracellular matrix proteins, and has been demonstrated to exhibit tumor-promoting and tumor-suppressive functions in different types of cancer. However, the function of LTBP2 in gastric cancer (GC) remains unknown. The aim of the present study was to investigate the expression and molecular function of LTBP2 in GC, and to evaluate its prognostic value for patients with GC. The results revealed that the expression of LTBP2 was upregulated in GC tissues and cell lines. Increased LTBP2 expression was associated with poor overall survival in patients with early-stage [tumor-node-metastasis (TNM) I/II] and late-stage (TNM III/IV) GC. Furthermore, silencing of LTBP2 effectively suppressed the proliferation, migration, invasion and epithelial-mesenchymal transition in GC cells. These results suggested that LTBP2 may be considered as a potential therapeutic target and a promising prognostic biomarker for human GC.

The prognostic role of pretreatment platelet-to-lymphocyte ratio as predictors in patients with colorectal cancer: a meta-analysis.

AIM: This meta-analysis was designed to analyze and evaluate the prognostic role of preoperative or pretreatment platelet-to-lymphocyte ratio (PLR) in patients with colorectal cancer (CRC). METHOD: We searched PubMed, Embase, Cochrane Library, Web of Science, China National Knowledge Infrastructure, Chinese Biomedical Literature Database and WanFang Database up to April 2016. RESULTS: A total of 16 studies (n = 5068 participants) were included for this meta-analysis. Elevated PLR has a close relationship with the poor overall survival of CRC, with the pooled hazard ratio being 1.88 (95% CI: 1.50, 2.36; p < 0.00001). CONCLUSION: This meta-analysis indicated that pretreatment PLR may be a cost-effective and noninvasive serum biomarker for poor prognosis for patients with CRC.

Preparation and copper ions adsorption properties of thiosemicarbazide chitosan from squid pens.

Chitosan was prepared by alkaline N-deacetylation of ß-chitin from squid pens. Thiosemicarbazide group was introduced to chitosan via formaldehyde-derived linkages, and thiosemicarbazide chitosan (TSFCS) with different degrees of substitution (DS) was synthesized. The DS values of TSFCS calculated by elemental analysis were 0.19, 0.36 and 0.63. The structure of the TSFCS was confirmed by elemental analysis, FTIR, XRD, TGA and SEM. The adsorption capacity of Cu(II) ions by TSFCS showed good correlation with the DS and pH (pH range 2.2-5.8). The maximum Cu(II) ions adsorption capacity of all three TSFCS samples reached 134.0mgg-1 at pH 3.6, but chitosan showed no adsorption at this pH. The adsorption equilibrium process of Cu(II) ions onto TSFCS was better described by the Langmuir model than the Freundlich isotherm model. Cu(II) ions adsorbed by TSFCS could be released using 0.01M Na2EDTA and the adsorption capacity could retain above 80% after five adsorption-desorption cycles. TSFCS exhibited good potential for heavy metal removal because of its high adsorption capacity at the low pH.

The pretreatment thrombocytosis may predict prognosis of patients with colorectal cancer: a systematic review and meta-analysis.

AIM: Recently, several studies have reported that thrombocytosis may be associated with the poor prognosis of colorectal cancer (CRC). Nevertheless, their conclusions were still controversial. Results & methodology: We searched PubMed, Embase, Cochrane Library and Web of Science up to April 2016. A total of 30 studies including 9129 patients were included in this meta-analysis. Thrombocytosis had a close relationship with the poor overall survival of CRC compared with normal platelet counts, with the pooled hazard ratios being 1.89 (95% CI: 1.45-2.47; p < 0.00001) and 1.83 (95% CI: 1.33-2.53; p = 0.0002), with univariate and multivariate analyses, respectively. DISCUSSION & CONCLUSION: This meta-analysis indicated that thrombocytosis may be a cost-effective and noninvasive indicator for poor prognosis of patients with CRC, especially for overall survival.