Computational design of transmembrane pores.

Transmembrane channels and pores have key roles in fundamental biological processes1 and in biotechnological applications such as DNA nanopore sequencing2-4, resulting in considerable interest in the design of pore-containing proteins. Synthetic amphiphilic peptides have been found to form ion channels5,6, and there have been recent advances in de novo membrane protein design7,8 and in redesigning naturally occurring channel-containing proteins9,10. However, the de novo design of stable, well-defined transmembrane protein pores that are capable of conducting ions selectively or are large enough to enable the passage of small-molecule fluorophores remains an outstanding challenge11,12. Here we report the computational design of protein pores formed by two concentric rings of α-helices that are stable and monodisperse in both their water-soluble and their transmembrane forms. Crystal structures of the water-soluble forms of a 12-helical pore and a 16-helical pore closely match the computational design models. Patch-clamp electrophysiology experiments show that, when expressed in insect cells, the transmembrane form of the 12-helix pore enables the passage of ions across the membrane with high selectivity for potassium over sodium; ion passage is blocked by specific chemical modification at the pore entrance. When incorporated into liposomes using in vitro protein synthesis, the transmembrane form of the 16-helix pore-but not the 12-helix pore-enables the passage of biotinylated Alexa Fluor 488. A cryo-electron microscopy structure of the 16-helix transmembrane pore closely matches the design model. The ability to produce structurally and functionally well-defined transmembrane pores opens the door to the creation of designer channels and pores for a wide variety of applications.

Application of validated migration models for the risk assessment of styrene and acrylonitrile in ABS plastic toys.

With styrene and acrylonitrile in ABS plastic toys as examples, this paper introduces to the development of a systematic strategy for studying the chemical migration risk in toys. The approach, included the detection method, establishment of migration model, model verification, and the practical application of the model in risk assessment. First, simple and sensitive methods for detecting analyte residues and migration were developed by headspace GC-MS. Then, the migration models were established based on the migration data from 5 min to 168 h and verified using 11 ABS samples. The results showed that the predicted values of the models and the experimental values had a good fit (RMSE=0.10-8.72 %). Subsequently, the migration of analytes in 94 ABS toys was predicted with these models at specific migration times. The daily average exposure level to styrene and acrylonitrile were estimated for children (3 months to 3 years). At last, the migration models reasonably predicted that the cancer risk of styrene and acrylonitrile in ABS toys were 1.6 × 10-8-1.4 × 10-6 and 3.1 × 10-8-1.6 × 10-6, respectively. This research contributes to promote toy safety and child health by enriching migration models and risk assessments.

Electromembrane Extraction and Dual-Channel Nanoelectrospray Ionization Coupled with a Miniature Mass Spectrometer: Incorporation of a Dicationic Ionic Liquid-Induced Charge Inversion Strategy.

Green analytical chemistry aims at developing analytical methods with minimum use and generation of hazardous substances for the protection of human health and the environment. To address this need, a green analytical protocol has been developed for the analysis of anionic compounds integrating electromembrane extraction (EME), dual-channel nanoelectrospray ionization (nanoESI), and a miniature mass spectrometer. Haloacetic acids (HAAs) have attracted considerable public concern due to their adverse effects on human health and were selected as model analytes for method development. A flat membrane EME device was developed and assembled in-house. Optimization of fundamental operational parameters was performed using single-factor test and response surface methodology. Both the EME acceptor phase and an imidazolium-based dicationic ionic liquid (DIL), 1,1-bis(3-methylimidazolium-1-yl) butylene difluoride (C4(MIM)2F2), were subjected to dual-channel nanoESI and miniature mass spectrometry analysis based on a charge inversion strategy, where positively charged complexes were formed. Enhancement in signal intensity by as much as 2 magnitudes was achieved in the positive-ion mode compared to the negative-ion mode in the absence of the dicationic ion-pairing agent. The developed protocol was validated, obtaining good recoveries ranging from 82.7 to 109.9% and satisfactory sensitivity with limits of detection (LODs) and quantitation (LOQs) in the ranges of 1-5 and 2-10 µg/L, respectively. The greenness of the analytical procedure was assessed with a calculated score of 0.71, indicating a high degree of greenness. The developed method was applied to the analysis of real environmental or municipal water samples (n = 16), exhibiting appealing potential for outside-the-laboratory applications.

Wood preservatives in children's wooden toys from China: Distribution, migration, oral exposure, and risk assessment.

A total of 90 wooden toys were collected, and six wood preservatives (chlorophenols and lindane) were analyzed by using gas chromatography-tandem mass spectrometry to assess the exposure risk of children to wood preservatives through oral contact with wooden toys. The detection rates of six preservatives ranged from 2.2% to 22.2%. The contents of the preservatives ranged from 0.6 µg/kg to 9.6 µg/kg. 2,4-Dichlorophenol (2,4-DCP) and 2,4,6-trichlorophenol (2,4,6-TCP) had higher detection rates and contents than other preservatives. Thus, their migration behaviors from toys to saliva were further investigated. In 11 positive samples, the max migration ratios of 2,4-DCP and 2,4,6-TCP ranged from 7.1% to 20.3% and from 11.1% to 24.8%, respectively. For children aged 3-36 months, the daily average 2,4-DCP exposure level associated with wooden toys ranged from 2.7 pg/(kg day) to 46.9 pg/(kg day), and the daily average 2,4,6-TCP exposure ranged from 3.6 pg/(kg day) to 69.4 pg/(kg day). The contribution to exposure provided by the saliva mobilization pathway was more than that provided by the ingestion of scraped-off toys, and the exposure level of 2,4,6-TCP was greater than that of 2,4-DCP. The max hazard quotient for 2,4-DCP was 1.9 × 10-4, and the max cancer risk for 2,4,6-TCP was 1.2 × 10-9. The above results indicated that although wood preservatives were distributed in wooden toys, exposure arising from directly mouthing these materials currently does not pose unacceptable risks to children.

Migration regularity of six preservatives from wooden children's products to saliva and sweat based on microstructure-related migration models.

Wood preservatives in wooden children's products (WCPs) may migrate into children's body through oral and dermal exposure, resulting in a potential health risk. In this paper, a systematic investigation on the migration regularity of lindane and five chlorophenols preservatives from WCPs to saliva and sweat was introduced. Migration models were established based on the abundant migration data among different time periods (2 min-96 h). Wood exhibited complex porosity for various species, resulting in the deviation of the migration of preservatives from different samples. By introducing a correction coefficient (f) calculated based on the sample microstructure (specific surface area and pore distribution), the predicted values of correcting migration model matched better with the experimental data. Migration data indicated that exposure risks should be noticed when children were in contact with WCPs because a considerable quantity of preservatives migrated into the body in a short time. This work is expected to play a role in the formulation of chemical limit standards to promote the safety of WCPs, and may serve ideas as basis for the migration research of chemicals in wooden products.

Alterations of the oral microbiome in patients treated with the Invisalign system or with fixed appliances.

INTRODUCTION: Although the Invisalign system has been used widely in recent years, the influences of this treatment on the oral microbiome and whether or not this influence is different from that of fixed appliances is still unknown. In this study, we investigated the changes in the oral microbiome in patients treated with the Invisalign system or with fixed appliances. METHODS: Fifteen subjects were enrolled, comprising 5 fixed appliance patients, 5 Invisalign patient, and 5 healthy controls. Saliva samples were collected, and high-throughput pyrosequencing was performed based on the 16S rRNA gene. RESULTS: Both fixed and Invisalign orthodontic treatments resulted in dysbiosis of the oral microbiome. Firmicutes and TM7 at the phyla level and Neisseria at the genus level displayed statistically significant differences between the 2 orthodontic groups. The effect of these changes with microbiome on oral health was inconsistent. The inferred microbial function of the Invisalign group suggested this group was more predisposed to periodontal diseases. CONCLUSION: The influence of the Invisalign system on the oral microbiome was no better for oral health compared with fixed appliances. The convenience of maintaining oral hygiene rather than changes in the oral microbiome may be the underlying reason for the performance of the Invisalign system on oral health.

Nitrogen-driven stem elongation in poplar is linked with wood modification and gene clusters for stress, photosynthesis and cell wall formation.

BACKGROUND: Nitrogen is an important nutrient, often limiting plant productivity and yield. In poplars, woody crops used as feedstock for renewable resources and bioenergy, nitrogen fertilization accelerates growth of the young, expanding stem internodes. The underlying molecular mechanisms of nitrogen use for extension growth in poplars are not well understood. The aim of this study was to dissect the nitrogen-responsive transcriptional network in the elongation zone of Populus trichocarpa in relation to extension growth and cell wall properties. RESULTS: Transcriptome analyses in the first two internodes of P. trichocarpa stems grown without or with nitrogen fertilization (5 mM NH4NO3) revealed 1037 more than 2-fold differentially expressed genes (DEGs). Co-expression analysis extracted a network containing about one-third of the DEGs with three main complexes of strongly clustered genes. These complexes represented three main processes that were responsive to N-driven growth: Complex 1 integrated growth processes and stress suggesting that genes with established functions in abiotic and biotic stress are also recruited to coordinate growth. Complex 2 was enriched in genes with decreased transcript abundance and functionally annotated as photosynthetic hub. Complex 3 was a hub for secondary cell wall formation connecting well-known transcription factors that control secondary cell walls with genes for the formation of cellulose, hemicelluloses, and lignin. Anatomical and biochemical analysis supported that N-driven growth resulted in early secondary cell wall formation in the elongation zone with thicker cell walls and increased lignin. These alterations contrasted the N influence on the secondary xylem, where thinner cell walls with lower lignin contents than in unfertilized trees were formed. CONCLUSION: This study uncovered that nitrogen-responsive elongation growth of poplar internodes is linked with abiotic stress, suppression of photosynthetic genes and stimulation of genes for cell wall formation. Anatomical and biochemical analysis supported increased accumulation of cell walls and secondary metabolites in the elongation zone. The finding of a nitrogen-responsive cell wall hub may have wider implications for the improvement of tree nitrogen use efficiency and opens new perspectives on the enhancement of wood composition as a feedstock for biofuels.

[The study of new soft one-piece PHEMA keratoprosthesis implanted into alkali burned rabbits cornea].

OBJECTIVE: This study was to evaluate clinical results of two types soft one-piece keratoprosthesis (KPros) made of PHEMA implanted to alkali burned rabbit corneas. METHODS: Experimental study. Twelve pieces KPros were implanted alkali burned rabbit corneas(type I and II, 6 pieces respectively). The examinations including slit-lamp, fundus photography,B-ultrasound and ultrasound biomicroscopy (UBM) were carried out. RESULTS: All Kpros were stable and no complications happened including conjunctiva flap dehiscence, aqueous leak and infection. IOP were normal by finger touch. B-ultrasound show no retina detachment and UBM show synechia in most animals (10 case). Retroprosthetic membrane happened in type I (3 case) . Visible conjunctiva hyperplasia covering the optical region happened in all KPros. CONCLUSIONS: (1) The same material, integrated design, maybe eliminating the material interface problem. (2)Simple surgical skills. (3)IOP measurement is possible because of soft material. (4)KPro I do not need removal of the lens and vitrectomy. The projecting portion of KPro II may reduce the incidence of proliferative membrane. Long term effects need more cases and further observation.

Nontargeted analysis and comparison strategies for volatile and semivolatile substances in toys made of different materials.

A nontargeted broad-spectrum analysis method for unknown volatile and semivolatile substances in toys was established by gas chromatography-Orbitrap high-resolution mass spectrometry. Based on the NIST spectrum library, unknown substances could be accurately identified by comprehensive scoring, retention index, chemical ionization, and fine comparison of ion fragments. For substances not included in the library, the molecular formulas of unknown substances were retrieved through online compound databases. Possible structural formulas were verified by high-resolution spectra and fragmentation mechanisms. Taking teether toys as an example, the substances differences of products made of different materials were compared through the digitization of chemical composition. Specifically, 59 substances were identified in 50 teether toys. The toys made of two different materials each had their own substance distribution, and the types and quantities of substances in thermoplastic polyurethanes samples were more than those in silicone samples. Substances with high risk included phenol, N-methylaniline, cyclohexanone, and 4-tert-amylphenol. This work can serve as a reference for the identification of unknown substances in toys and other products, as well as for the comparison the chemical composition of products made of different materials. Thus, this work has positive significance in promoting the quality and safety of toys and reducing chemical harm to children.

N-fertilization has different effects on the growth, carbon and nitrogen physiology, and wood properties of slow- and fast-growing Populus species.

To investigate how N-fertilization affects the growth, carbon and nitrogen (N) physiology, and wood properties of poplars with contrasting growth characteristics, slow-growing (Populus popularis, Pp) and fast-growing (P. alba×P. glandulosa, Pg) poplar saplings were exposed to different N levels. Above-ground biomass, leaf area, photosynthetic rates (A), instantaneous photosynthetic nitrogen use efficiency (PNUE (i)), chlorophyll and foliar sugar concentrations were higher in Pg than in Pp. Foliar nitrate reductase (NR) activities and root glutamate synthase (GOGAT) activities were higher in Pg than in Pp as were the N amount and NUE of new shoots. Lignin contents and calorific values of Pg wood were less than that of Pp wood. N-fertilization reduced root biomass of Pg more than of Pp, but increased leaf biomass, leaf area, A, and PNUE(i) of Pg more than of Pp. Among 13 genes involved in the transport of ammonium or nitrate or in N assimilation, transcripts showed more pronounced changes to N-fertilization in Pg than in Pp. Increases in NR activities and N contents due to N-fertilization were larger in Pg than in Pp. In both species, N-fertilization resulted in lower calorific values as well as shorter and wider vessel elements/fibres. These results suggest that growth, carbon and N physiology, and wood properties are more sensitive to increasing N availability in fast-growing poplars than in slow-growing ones, which is probably due to prioritized resource allocation to the leaves and accelerated N physiological processes in fast-growing poplars under higher N levels.

Lamp1 mediates lipid transport, but is dispensable for autophagy in Drosophila.

The endolysosomal system not only is an integral part of the cellular catabolic machinery that processes and recycles nutrients for synthesis of biomaterials, but also acts as signaling hub to sense and coordinate the energy state of cells with growth and differentiation. Lysosomal dysfunction adversely influences vesicular transport-dependent macromolecular degradation and thus causes serious problems for human health. In mammalian cells, loss of the lysosome associated membrane proteins LAMP1 and LAMP2 strongly affects autophagy and cholesterol trafficking. Here we show that the previously uncharacterized Drosophila Lamp1 is a bona fide ortholog of vertebrate LAMP1 and LAMP2. Surprisingly and in contrast to lamp1 lamp2 double-mutant mice, Drosophila Lamp1 is not required for viability or autophagy, suggesting that fly and vertebrate LAMP proteins acquired distinct functions, or that autophagy defects in lamp1 lamp2 mutants may have indirect causes. However, Lamp1 deficiency results in an increase in the number of acidic organelles in flies. Furthermore, we find that Lamp1 mutant larvae have defects in lipid metabolism as they show elevated levels of sterols and diacylglycerols (DAGs). Because DAGs are the main lipid species used for transport through the hemolymph (blood) in insects, our results indicate broader functions of Lamp1 in lipid transport. Our findings make Drosophila an ideal model to study the role of LAMP proteins in lipid assimilation without the confounding effects of their storage and without interfering with autophagic processes.Abbreviations: aa: amino acid; AL: autolysosome; AP: autophagosome; APGL: autophagolysosome; AV: autophagic vacuole (i.e. AP and APGL/AL); AVi: early/initial autophagic vacuoles; AVd: late/degradative autophagic vacuoles; Atg: autophagy-related; CMA: chaperone-mediated autophagy; Cnx99A: Calnexin 99A; DAG: diacylglycerol; eMI: endosomal microautophagy; ESCRT: endosomal sorting complexes required for transport; FB: fat body; HDL: high-density lipoprotein; Hrs: Hepatocyte growth factor regulated tyrosine kinase substrate; LAMP: lysosomal associated membrane protein; LD: lipid droplet; LDL: low-density lipoprotein; Lpp: lipophorin; LTP: Lipid transfer particle; LTR: LysoTracker Red; MA: macroautophagy; MCC: Manders colocalization coefficient; MEF: mouse embryonic fibroblast MTORC: mechanistic target of rapamycin kinase complex; PV: parasitophorous vacuole; SNARE: soluble N-ethylmaleimide sensitive factor attachment protein receptor; Snap: Synaptosomal-associated protein; st: starved; TAG: triacylglycerol; TEM: transmission electron microscopy; TFEB/Mitf: transcription factor EB; TM: transmembrane domain; tub: tubulin; UTR: untranslated region.

Enriched Au nanoclusters with mesoporous silica nanoparticles for improved fluorescence/computed tomography dual-modal imaging.

OBJECTIVES: Au nanoclusters (AuNCs) have been used widely in fluorescence bio-imaging because of their good fluorescence, small particle size and non-cytotoxicity. AuNCs are also efficient in computed tomography (CT) imaging. Hence, a dual-modal imaging probe can be constructed without any complicated modification processes by exploiting the excellent performance of AuNCs. In the present study, AuNCs were enriched with mesoporous silica nanoparticles (MSNs) to obtain enhanced fluorescence/CT dual-modal imaging, which was capable of acquiring more imaging information for diseases compared with single-mode imaging. MATERIALS AND METHODS: Biocompatible bovine serum albumin (BSA)-capped AuNCs were prepared and loaded into amine-functionalized MSNs to form MSN@AuNCs. BSA-AuNCs, MSNs, and MSN@AuNCs were characterized by ultraviolet-visible (UV-vis) spectra, transmission electron microscopy (TEM), fluorescence spectra, and zeta potential. CT imaging was recorded using micro-CT scanning. Fluorescence imaging was measured using confocal laser scanning microscopy and flow cytometry. RESULTS: The prepared AuNCs and MSNs possessed good properties as previously reported. The fluorescence intensity and CT value of the AuNCs were enhanced after being enriched with MSNs. The nanoparticles were both non-cytotoxic. Confocal laser scanning microscopy and flow cytometry indicated that MSN@AuNCs in CAL-27 cells showed improved fluorescence imaging compared with simple AuNCs at the same concentration. CONCLUSIONS: The results revealed that the strategy of enriching AuNCs with MSNs can obtain highly sensitive fluorescence/CT dual-modal imaging, which indicated the potential of this nanoparticle in the diagnosis and treatment of disease.

Conducting polymer nanomaterials: electrosynthesis and applications.

Conducting polymers (CPs) have been extensively studied and widely applied in various organic devices. To improve the performances or extend the functions of the devices, CPs usually have to be nanostructured. Electrosynthesis provides an effective and convenient one-step approach to CP nanomaterials. The resulting materials are usually oriented on the electrode surfaces and their properties are easy to be controlled. This critical review focuses on the syntheses of CP nanostructures and nanocomposites by electrochemical polymerization. The applications of the nanomaterials in organic devices such as sensors, actuators and memory devices also will be discussed (111 references).

Evaluation of the bond strength of chlorhexidine incorporated into the adhesive system composition: A PRISMA guided meta-analysis.

BACKGROUND/PURPOSE: Composite resin is currently the most widely used dental restoration material. Previous studies have demonstrated that the application of Chlorhexidine (CHX) on the dentin surface after acid etching can result in an improvement in the integrity and stability of tooth restoration through time. In order to better understand whether CHX can help improve the stability of the resin-dentin bond strength, in this study, a comprehensive review of the effect of adding CHX to the adhesive system on the stability of immediate and long-term resin-dentin bond strength was conducted. MATERIALS AND METHODS: This article was written in accordance with the PRISMA Statement and is registered on the International Prospective Register of Systematic Reviews (registration number CRD42018084962). Six electronic databases including PubMed, Embase, Cochrane library, ISI Web of Science, ClinicalTrials.gov and China National Knowledge Infrastructure (CNKI) were searched up to October, 2018. Ten articles were selected from 340 possible eligible articles for meta-analysis, and 41 sets of data were analyzed in the meta-analysis. RESULTS: The results indicated that the use of 0.1% and 0.2% CHX does not adversely affect the immediate bond strength (p > 0.05), but both 0.1% and 0.2% CHX increased bond strength compared with the control group over 12 months (p < 0.05). However, this trend does not represent a longer period of aging. CONCLUSION: In these in vitro clinical trials, CHX incorporated into the bonding systems maintained the stability of bond strength.

Pegylated recombinant human granulocyte colony-stimulating factor regulates the immune status of patients with small cell lung cancer.

BACKGROUND: Small cell lung cancer (SCLC) is an aggressive disease involving immunodeficiency for which chemotherapy is the standard treatment. Pegylated recombinant human granulocyte colony-stimulating factor (PEG-rhG-CSF) is widely used for primary or secondary prophylaxis of febrile neutropenia (FN) in chemotherapy. However, whether PEG-rhG-CSF influences immune cells, such as lymphocytes, remains unclear. METHODS: A total of 17 treatment-naïve SCLC patients were prospectively enrolled and divided into the PEG-rhG-CSF and control groups according to their FN risk. Longitudinal sampling of peripheral blood was performed before, after and 4-6 days after the first cycle of chemotherapy. Flow cytometry was used to assess lymphocyte subsets, including CD3+ T, CD4+ T, CD8+ T, NK, and B cells. The diversity and clonality of the T-cell receptor (TCR) repertoire was analyzed by next-generation sequencing. RESULTS: In the PEG-rhG-CSF group, the proportions of CD3+ T and CD4+ T cells had increased significantly (P = 0.002, P = 0.020, respectively), whereas there was no increase in CD8+ T cells. Further, TCR diversity increased (P = 0.009) and clonality decreased (P = 0.004) significantly after PEG-rhG-CSF treatment. However, these factors showed opposite trends before and after chemotherapy. Vß and Jß gene fragment types, which determine TCR diversity, were significantly amplified in the PEG-rhG-CSF group. The change in TCR diversity was significantly correlated with changes in the CD3+ T or CD4+ T cell proportions, but not the CD8+ T cell proportion. CONCLUSIONS: PEG-rhG-CSF regulates the immune status of SCLC patients; CD4+ T cells may be the main effector cells involved in this process. These findings may optimize the treatment of SCLC. KEY POINTS: PEG-rhG-CSF regulates SCLC immunity. PEG-rhG-CSF increased CD3+ T and CD4+ T cell proportions. PEG-rhG-CSF increased TCR diversity and decreased clonality in peripheral blood. Change in TCR diversity were correlated with CD3+ T or CD4+ T changes.

Suspect screening of 200 hazardous substances in plastic toys using ultra-high-performance liquid chromatography-hybrid quadrupole time-of-flight mass spectrometry.

There is an urgent need for the development of efficient and comprehensive analytical methods for organic chemical compounds due to their increasing number and diversity in children's toy products. The presence of these chemicals in toys poses an extreme risk for the health and development of children. In this study, an analytical methodology has been developed using ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS). This hybrid instrumentation together with an in-house accurate-mass database and a mass spectral library, allows for wide-scope screening and identification of hazardous substances in plastic toys. A total number of 200 compounds belonging to eight chemical families were investigated, such as coloring agents, plasticizers, fragrance allergens, nitrosamines, primary aromatic amines, flame retardants, perfluorinated compounds, and endocrine disruptors. Following a straightforward and efficient dissolution/precipitation method for sample preparation, chemical screening and confirmation were conducted by comparing the experimentally measured exact mass, retention time, and isotopic pattern with the accurate-mass database and by matching the acquired MS/MS spectra against the mass spectral library. The matrix effect, linearity, sensitivity, precision, and recovery of the proposed method were properly evaluated. The obtained limits of detection (LODs) and quantitation (LOQs) were in the range of 0.01-0.98 mg kg-1 and 0.03-2.99 mg kg-1, respectively. The applicability of the developed protocol was verified through the analysis of 55 real plastic toy products.

The potential effect of Bruton's tyrosine kinase in refractory periapical periodontitis.

To determine the expression of Bruton's tyrosine kinase (BTK) in refractory periapical periodontitis and analyze the relationship between BTK and bone resorption in refractory periapical periodontitis. The mechanism of bone resorption is also discussed. The OneArray Plus expression microarray was used to screen for genes related to refractory periapical periodontitis. Real-time PCR was used to detect the expression of BTK in refractory periapical periodontitis tissues. A model of periapical periodontitis was established by sealing E.faecalis into the pulp of rats. To establish a model of E.faecalis LTA infection of osteoclasts, the relationship between BTK and bone destruction during refractory periapical periodontitis was analyzed. OneArray Plus expression microarray results showed that we found that the expression of 1787 genes in the two samples was different. After validating these samples, we found that BTK was closely related to refractory periapical periodontitis. The results showed that the expression of BTK in refractory periapical periodontitis tissues was higher than that in normal tissues. Immunohistochemistry, enzyme histochemistry and real-time PCR showed that the BTK expression curve in the experimental model resembled a reverse V shape from week 1 to week 4. Osteoclasts were cultured in vitro and treated with E. faecalis LTA. The expression of BTK in the E. faecalis model was greater than that in the control group. BTK played an important role in the progression of refractory periapical periodontitis.

Optimization of multi-residue method for targeted screening and quantification of 216 harmful chemicals in plastic children's toys by gas chromatography-tandem mass spectrometry analysis.

Herein, we report a multi-residue method based on dissolution-precipitation extraction combined with gas chromatography-tandem mass spectrometry (GC-MS/MS) analysis for targeted screening and quantification of 216 harmful chemicals in plastic children's toys. We established a spectrogram database containing the first-order mass spectra of all analytes and their second-order mass spectra at different collision energies. Good separation was attained in less than 60 min. As a simple and low-cost extraction method, dissolution-precipitation exhibited good recoveries for the analytes in the plastic samples. The matrix-matched standard curves were accurate for analyte quantification in specific plastics due to the solvent effects and matrix effects. The limits of quantification range were 0.1 mg/kg to 10 mg/kg. A wide linear range of 0.1-200 mg/kg was also observed, with r2≥0.9924. The average recoveries ranged from 63.9% to 137.6%, and the relative standard deviation (n = 6) varied from 0.6% to 13.2%. Finally, 31 ABS actual toys and 30 PVC actual toys were detected and 20 analytes with contents ranging from 0.4 mg/kg to 172 mg/kg and 30 analytes from 0.8 mg/kg to 141,030 mg/kg were discovered respectively.

[Implantation of modified polyhydroxyethyl methacrylate-polymethyl methacrylate keratoprostheses in rabbit and monkey corneas].

OBJECTIVE: To investigate the biocolonization of polyhydroxyethyl methacrylate (PHEMA) sponge with cornea tissue and evaluate the therapeutic effects of modified porous PHEMA-PMMA (polymethyl methacrylate) Keratoprostheses (KPro) on rabbit and monkey corneas. METHODS: The KPro were made using two-stage polymerization combined with mechanical cutting. The experiment was divided into two groups. In the control group (A group), ten normal rabbit eyes received lamellar implantation of PHEMA sponges. The sponges were obtained 2 weeks, 1, 2, 3 and 4 months after operation. The cell proliferation and neovascularization inside the sponges were observed using light and transmission electron microscopy (TEM) and immunohistochemistry. In the experimental group (B group), the porous PHEMA-PMMA KPros were inserted into the lamellar pockets of ten rabbit corneas and two monkey corneas (stage I operation). The healing process was investigated by slit-lamp microscopy. The anterior lamellar cornea tissues were removed 3 months after surgery, exposing the underneath transparent core (stage II operation). The operated eyes were then followed up for 3 - 6 months. RESULTS: No complication was observed in A group. Under the light microscope, fibroblasts started to grow into the cornea 2 weeks after operation; lots of cells, accompanied with new blood vessels, invaded into the cornea 2 - 3 months after surgery. Invading cells of sponge, as well as keratocyte, were positive for vimentin. Under the electron microscope, the invading cells looked healthy and were surrounded by extracellular matrix and collagen. In B group, eight rabbit eyes which have received KPro implantation, anterior lamellar cornea melting happened in two eyes after the stage I operation. The remaining six corneas retained their central cores during observation after the stage II operation. Two monkey operated eyes were found no complication throughout the whole follow-up. CONCLUSIONS: The PHEMA sponge can obtain a tight fusion with the host cornea. The modified PHEMA-PMMA KPros have obtained a relatively stable therapeutic results after implantation into animal corneas.

Microporous Organic Polymers Based on Hyper-Crosslinked Coal Tar: Preparation and Application for Gas Adsorption.

Hyper-crosslinked polymers (HCPs) are promising materials for gas capture and storage, but high cost and complicated preparation limit their practical application. In this paper, a new type of HCPs (CTHPs) was synthesized through a one-step mild Friedel-Crafts reaction with low-cost coal tar as the starting material. Chloroform was utilized as both solvent and crosslinker to generate a three-dimensional crosslinked network with abundant micropores. The maximum BET surface area of the prepared CTHPs could reach up to 929 m2 g-1 . Owing to the high affinity between the heteroatoms on the coal-tar building blocks and the CO2 molecules, the adsorption capacity of CTHPs towards CO2 reached up to 14.2 wt % (1.0 bar, 273 K) with a high selectivity (CO2 /N2 =32.3). Furthermore, the obtained CTHPs could adsorb 1.27 wt % H2 at 1.0 bar and 77.3 K, and also showed capacity for the capture of high organic vapors at room temperature. In comparison with other reported porous organic polymers, CTHPs have the advantages of low-cost, easy preparation, and high gas-adsorption performance, making them suitable for mass production and practical use in the future.