[Preparation technology comparison and performance evaluation of different protein A affinity chromatographic materials].

Protein A affinity chromatographic materials are widely used in clinical medicine and biomedicine because of their specific interactions with immunoglobulin G (IgG). Both the characteristics of the matrix, such as its structure and morphology, and the surface modification method contribute to the affinity properties of the packing materials. The specific, orderly, and oriented immobilization of protein A can reduce its steric hindrance with the matrix and preserve its bioactive sites. In this study, four types of affinity chromatographic materials were obtained using agarose and polyglycidyl methacrylate (PGMA) spheres as substrates, and multifunctional epoxy and maleimide groups were used to fix protein A. The effects of the ethylenediamine concentration, reaction pH, buffer concentration, and other conditions on the coupling efficiency of protein A and adsorption performance of IgG were evaluated. Multifunctional epoxy materials were prepared by converting part of the epoxy groups of the agarose and PGMA matrices into amino groups using 0.2 and 1.6 mol/L ethylenediamine, respectively. Protein A was coupled to the multifunctional epoxy materials using 5 mmol/L borate buffer (pH 8) as the reaction solution. When protein A was immobilized on the substrates by maleimide groups, the agarose and PGMA substrates were activated with 25% (v/v) ethylenediamine for 16 h to convert all epoxy groups into amino groups. The maleimide materials were then converted into amino-modified materials by adding 3 mg/mL 3-maleimidobenzoyl-N-hydroxysuccinimide ester (MBS) dissolved in dimethyl sulfoxide (DMSO) and then suspended in 5 mmol/L borate buffer (pH 8). The maleimide groups reacted specifically with the C-terminal of the sulfhydryl group of recombinant protein A to achieve highly selective fixation on both the agarose and PGMA substrates. The adsorption performance of the affinity materials for IgG was improved by optimizing the bonding conditions of protein A, such as the matrix type, matrix particle size, and protein A content, and the adsorption properties of each affinity material for IgG were determined. The column pressure of the protein A affinity materials prepared using agarose or PGMA as the matrix via the maleimide method was subsequently evaluated at different flow rates. The affinity materials prepared with PGMA as the matrix exhibited superior mechanical strength compared with the materials prepared with agarose. Moreover, an excellent linear relationship between the flow rate and column pressure of 80 mL/min was observed for this affinity material. Subsequently, the effect of the particle size of the PGMA matrix on the binding capacity of IgG was investigated. Under the same protein A content, the dynamic binding capacity of the affinity materials on the PGMA matrix was higher when the particle size was 44-88 µm than when other particle sizes were used. The properties of the affinity materials prepared using the multifunctional epoxy and maleimide-modified materials were compared by synthesizing affinity materials with different protein A coupling amounts of 1, 2, 4, 6, 8, and 10 mg/mL. The dynamic and static binding capacities of each material for bovine IgG were then determined. The prepared affinity material was packed into a chromatographic column to purify IgG from bovine colostrum. Although all materials showed specific adsorption selectivity for IgG, the affinity material prepared by immobilizing protein A on the PGMA matrix with maleimide showed significantly better performance and achieved a higher dynamic binding capacity at a lower protein grafting amount. When the protein grafting amount was 15.71 mg/mL, the dynamic binding capacity of bovine IgG was 32.23 mg/mL, and the dynamic binding capacity of human IgG reached 54.41 mg/mL. After 160 cycles of alkali treatment, the dynamic binding capacity of the material reached 94.6% of the initial value, indicating its good stability. The developed method is appropriate for the production of protein A affinity chromatographic materials and shows great potential in the fields of protein immobilization and immunoadsorption material synthesis.

A novel lysosome-related prognostic signature associated with prognosis and immune infiltration landscape in oral squamous cell carcinoma.

Background: Predicting the outcome of oral squamous cell carcinoma (OSCC) is challenging due to its diverse nature and intricate causes. This research explores how lysosome-associated genes (LRGs) might forecast overall survival (OS) and correlate with immune infiltration in OSCC patients. Methods: We analyzed OSCC patients' LRGs' mRNA expression data and clinical details from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO). Through univariate Cox regression, we pinpointed LRGs with prognostic potential. A signature comprising 12 LRGs linked to prognosis was developed via the Least Absolute Shrinkage and Selection Operator (LASSO) in a training dataset. Patients were classified as higher or lower risk based on their risk scores, and the prognostic independence of the risk score was assessed using multivariate analysis. The model's robustness and precision were confirmed through bioinformatics in the GEO test set. Differential gene expression analysis between risk groups highlighted functional disparities, while various immune evaluation methods elucidated immune differences. Results: The prognostic framework utilized 12 LRGs (SLC46A3, MANBA, NEU1, SDCBP, BRI3, TMEM175, CD164, GPC1, SFTPB, TPP1, Biglycan (BGN) and TMEM192), showing that higher risk was associated with poorer OS. This set of genes independently predicted OS in OSCC, linking LRGs to cellular adhesion and extracellular matrix involvement. Initial assessments using ssGSEA and CIBERSORT suggested that the adverse outcomes in the higher-risk cohort may be tied to immune system deregulation. Conclusion: Twelve-LRGs signature has been identified for OSCC prognosis prediction, offering novel directions for lysosome-targeted therapies against OSCC.

Non-pharmacological interventions for preventing suicide attempts: A systematic review and network meta-analysis.

Suicide attempts can cause serious physical harm or death. It would be crucial to gain a better understanding of the comparative efficacy of non-pharmacological interventions. We aimed to identify which non-pharmacological interventions are more effective in preventing suicide attempts. PubMed, Web of Science, and EMBASE databases were searched systematically from their inception until 3 April 2023. To be eligible for inclusion, randomized controlled trials (RCTs) had to meet the following criteria: Participants were individuals who had suicidal ideation or a history of severe self-harm or attempted suicide. A network meta-analysis was performed using a random effects model to estimate the treatment effect of various non-pharmacological interventions. (PROSPERO registration number: CRD42023411393). We obtained data from 54 studies involving 17,630 participants. Our primary analysis found that Cognitive therapy (CT) (OR=0.19, 95%CI =0.04-0.81), Dialectical Behavior Therapy (DBT) (OR=0.37, 95%CI =0.13-0.97), Cognitive-behavioral therapy (CBT) (OR=0.42, 95%CI =0.17-0.99), and Brief intervention and contact (BIC) (OR=0.65, 95%CI=0.44-0.94) were superior to TAU (within the longest available follow-up time) in preventing suicide attempts, while other intervention methods do not show significant advantages over TAU. Secondary analysis showed that the two intervention measures (CT and BIC) were effective when follow-up time did not exceed 6 months, but there was no effective intervention measure with longer follow-up times. CT, DBT, CBT, and BIC have a better effect in preventing suicide attempts than other non-pharmacological interventions. Additional research is necessary to validate which interventions, as well as which combinations of interventions, are the most effective.

Observation of a Folded Dirac Cone in Heavily Doped Graphene.

Superlattice potentials imposed on graphene can alter its Dirac states, enabling the realization of various quantum phases. We report the experimental observation of a replica Dirac cone at the Brillouin zone center induced by a superlattice in heavily doped graphene with Gd intercalation using angle-resolved photoemission spectroscopy (ARPES). The replica Dirac cone arises from the (√3× âˆš3)R30° superlattice formed by the intervalley coupling of two nonequivalent valleys (e.g., the Kekulé-like distortion phase), accompanied by a bandgap opening. According to the findings, the replica Dirac band in Gd-intercalated graphene disappears beyond a critical temperature of 30 K and can be suppressed by potassium adsorption. The modulation of the replica Dirac band is primarily attributable to the residual frozen gas, which can act as a source of intervalley scattering at temperatures below 30 K. Our results highlight the persistence of the hidden Kekulé-like phase within the heavily doped graphene, enriching our current understanding of its replica Dirac Fermions.

Changes of maxillary central incisor and alveolar bone in Class II Division 2 nonextraction treatment with a fixed appliance or clear aligner: A pilot cone-beam computed tomography study.

INTRODUCTION: This retrospective clinical study investigated the clinical changes of maxillary central incisor and alveolar bone in Class II Division 2 nonextraction treatment with fixed appliances or clear aligners on the basis of cone-beam computed tomography. METHODS: Fifty-nine Chinese Han patients with similar demographic characteristics were collected from a conventional bracket group, a self-ligating bracket group, and a clear aligner group. All measurements about root resorption and alveolar bone thickness on the cone-beam computed tomography images were tested. Changes between pretreatment and posttreatment were evaluated by paired-sample t test. The variation among the 3 groups was compared by 1-way analysis of variance. RESULTS: The resistance center of the maxillary central incisor showed upward or forward movement, and the axial inclination was increased in 3 groups (P <0.0001). Root volume loss in the clear aligner group (23.68 ± 4.82 mm3) was significantly less than that in the fixed appliances group (28.24 ± 6.44 mm3 in the conventional bracket group, 28.17 ± 6.07 mm3 in the self-ligating bracket group) (P <0.05). All 3 groups showed a significant decrease in palatal alveolar bone and total bone thickness at all 3 levels at posttreatment. In contrast, labial bone thickness significantly increased except for crestal level l. Among the 3 groups, the clear aligner group had a prominent increase in labial bone thickness at the apical level (P = 0.0235). CONCLUSIONS: Clear aligner treatment for Class II Division 2 malocclusions could effectively reduce the incidence of fenestration and root resorption. Our findings will be beneficial to comprehensively understand the effectiveness of different appliances for Class II Division 2 malocclusions treatment.

Piezo1-mediated M2 macrophage mechanotransduction enhances bone formation through secretion and activation of transforming growth factor-ß1.

Macrophages are multifunctional immune system cells that are essential for the mechanical stimulation-induced control of metabolism. Piezo1 is a non-selective calcium channel expressed in multifarious tissues to convey mechanical signals. Here, a cellular model of tension was used to study the effect of mechanical stretch on the phenotypic transformation of macrophages and its mechanism. An indirect co-culture system was used to explore the effect of macrophage activation on bone marrow mesenchymal stem cells (BMSCs), and a treadmill running model was used to validate the mechanism in vivo for in vitro studies. p53 was acetylated and deacetylated by macrophages as a result of mechanical strain being detected by Piezo1. This process is able to polarize macrophages towards M2 and secretes transforming growth factor-beta (TGF-ß1), which subsequently stimulates BMSCs migration, proliferation and osteogenic differentiation. Knockdown of Piezo1 inhibits the conversion of macrophages to the reparative phenotype, thereby affecting bone remodelling. Blockade of TGF-ß I, II receptors and Piezo1 significantly reduced exercise-increased bone mass in mice. In conclusion, we showed that mechanical tension causes calcium influx, p53 deacetylation, macrophage polarization towards M2 and TGF-ß1 release through Piezo1. These events support BMSC osteogenesis.

Atf4 regulates angiogenic differences between alveolar bone and long bone macrophages by regulating M1 polarization, based on single-cell RNA sequencing, RNA-seq and ATAC-seq analysis.

In the repair of maxillofacial bone defects, autogenous craniofacial bone can often provide superior clinical results over long bone grafts. Most current studies have focused on the osteogenic differences between alveolar bone marrow (ABM) and long bone marrow (LBM), however, studies about the angiogenic differences between the two are currently lacking. We downloaded single-cell RNA sequencing (scRNA-seq) of mouse ABM and LBM respectively from the public database, and the data were processed by using Seurat package. CellphoneDB2 results showed that macrophages had the strongest interaction with mesenchymal stem cells (MSCs) and endothelial cells (ECs). ELISA results confirmed that ABM macrophages secreted a higher level of vascular endothelial growth factor A (Vegfa) compared to LBM macrophages, which further promoted angiogenesis of ECs and MSCs. Using SCENIC package, six key transcription factors (TFs) were identified to regulate the difference between ABM and LBM macrophages, and activating transcription factor 4 (Atf4) was confirmed to be more expressed in ABM macrophages by polymerase chain reaction (PCR) and western blot (WB), with predicted target genes including Vegfa. Besides, the result of scRNA-seq implied ABM macrophages more in M1 status than LBM macrophages, which was confirmed by the following experiments. From the results of another assay for transposase accessible chromatin sequencing (ATAC-seq) and RNA-seq about M1 macrophages, Atf4 was also confirmed to regulate the M1 polarization. So, we suspected that Atf4 regulated the different expression of Vegfa between ABM and LBM macrophages by activating M1 polarization. After knocking down Atf4, the expression of M1 polarization markers and Vegfa were downregulated and vasculogenic differences were eliminated, which were subsequently reversed by the addition of LPS/IFN-γ. Our study might provide a new idea to improve the success rate of autologous bone grafting and treatment of oral diseases.

Three-dimensional evaluation of the mandibular condyle in adults with various skeletal patterns.

Objective: Morphometric and morphological evaluation of the mandibular condyle in adults and to identify its correlation with skeletal malocclusion patterns. Methods: Cone-beam computed tomography scans of 135 adult patients were used in this study and classified into groups according to four criteria: (1) sex (male and female); (2) sagittal skeletal discrepancy (Class I, Class II, and Class III); (3) vertical skeletal discrepancy (hyperdivergent, normodivergent, and hypodivergent); and age (group 1 ≤ 20 years, 21 ≤ group 2 < 30, and group 3 ≥ 30 years). The morphometrical variables were mandibular condyle height and width, and the morphological variable was the mandibular condyle shape in coronal and sagittal sections. Three-dimensional standard tessellation language files were created using itk-snap (open-source software), and measurements were performed using Meshmixer (open-source software). Results: The mandibular condyle height was significantly greater (p < 0.05) in patients with class III malocclusion than in those with class I or II malocclusion; the mandibular condyle width was not significantly different among different sexes, age groups, and sagittal and vertical malocclusions. There were no statistical associations between various mandibular condyle shapes and the sexes, age groups, and skeletal malocclusions. Conclusions: The condylar height was greatest in patients with class III malocclusion. The condylar height and width were greater among males than in females. The mandibular condyle shapes observed in sagittal and coronal sections did not affect the skeletal malocclusion patterns.

Associations between mixed urinary phenols and parabens metabolites and bone mineral density: Four statistical models.

Phenols and parabens widely exist in personal care and consumer products and have been proved to be endocrine disrupting chemicals that could disturb bone metabolism. The current studies focusing on the associations between phenols and parabens with bone mineral density (BMD) drew contradictory conclusions. Considering the bias might be due to not considering the effects of mixed exposure, we conducted the first cross-sectional study to investigate the associations of both single and mixed metabolites of phenols and parabens with BMD in three populations by setting up four models: generalized linear regression model (GLM), weighted quantile sum (WQS) regression model, quantile g-computation (qgcomp) model and Bayesian kernel machine regression (BKMR) model, based on the US National Health and Nutrition Examination Survey (NHANES) database. We found that the association between the mixtures and total femur BMD in men was significantly negative. Bisphenol A (BPA) was shown to play the most important role in this negative association in the BKMR model, and this negative association was also confirmed in the GLM model with ß coefficient (95% CI) being -0.02 (-0.04, -0.01). The relationships between the mixtures and femoral neck and trochanter BMD in postmenopausal women were significantly positive. Benzophenone-3 (BP-3) played the most significant role in the positive association with trochanter BMD, as confirmed by the WQS, qgcomp and BKMR models, and this positive association was also verified by the GLM model with ß coefficient (95% CI) being 0.01 (0.00, 0.02). In conclusion, the association between the mixed phenols and parabens and BMD was negative in men while was positive in postmenopausal women, which was gender-specific. This study might provide new ideas for the prevention and treatment of osteoporosis and the control of personal care and consumer products containing phenols and parabens in the future.

Association between mixed aldehydes and bone mineral density based on four statistical models.

Aldehydes as an environmental pollutant may lead to oxidative stress, which is an important mechanism in the development of osteoporosis. This suggests a possible link between aldehyde exposure and osteoporosis. Considering the mixed nature of aldehyde exposure and the interactions between different aldehydes, we explored for the first time the associations between mixed six aldehydes (benzaldehyde, butyraldehyde, heptanal, hexanal, isovaleraldehyde, and propionaldehyde) and BMD in three populations (men, premenopausal women, and postmenopausal women) by applying four statistical models: quantile g-computation (qgcomp) model, Bayesian kernel machine regression (BKMR) model, generalized linear regression model (GLM), and generalized additive model (GAM), based on the National Health and Nutrition Examination Survey (NHANES) 2013-2014. We found that mixed aldehydes could significantly reduce BMD in men, with hexanaldehyde and propanaldehyde having the greatest negative qgcomp model and BKMR model weights, also confirmed by GLM. The associations between isopentanaldehyde and propanaldehyde and femoral BMD in men were non-linear and had threshold effects as derived from the BKMR model and GAM. The associations turned positive when the concentrations of isopentanaldehyde and propanaldehyde exceeded their respective inflection points. To conclude, our study might provide new ideas for the prevention and treatment of osteoporosis, and hexanaldehyde and propanaldehyde should be more regulated to prevent osteoporosis.

Comparison of vacuum suction ureteroscopic laser lithotripsy and traditional ureteroscopic laser lithotripsy for impacted upper ureteral stones.

PURPOSE: To compare a novel vacuum suction ureteroscopic laser lithotripsy (VS-URS) with traditional ureteroscopic laser lithotripsy (T-URS) for impacted upper ureteral stones and to better define the potential benefits of VS-URS. METHODS: Between May 2019 and March 2021, 158 patients with impacted upper ureteral stones underwent ureteroscopic holmium-YAG laser lithotripsy. Of these, 76 underwent VS-URS and 82 underwent T-URS. In VS-URS procedures, the vacuum suction device is composed of a 5F ureteral catheter and a tee joint. The ureteral catheter is linked to the vacuum aspirator by the sidearm of the tee joint, and a 200 µm fiber is inserted through the tee joint and the ureteral catheter into the stone site for lithotripsy. RESULTS: When compared to the T-URS group, the VS-URS group had a shorter mean operation time (38.18 ± 6.37 min vs. 46.65 ± 5.66 min; P = 0.000), lower fever rate (3.9% vs. 14.6%; P < 0.022), less stone retropulsion (5.3% vs. 18.3%; P = 0.012), lower extra management rate (6.58% vs. 21.95%; P = 0.006), and a higher stone-free rate of the first postoperative day (88.2% vs. 72.0%; P = 0.011). There were no significant differences in stone-free rates 1 month after surgery between groups (94.7% vs. 92.7%; P = 0.748). CONCLUSIONS: VS-URS is an effective modality for impacted upper ureteral stones, and has a shorter operating time, lower fever rate, less stone retropulsion, and a higher primary stone-free rate compared with T-URS.

Orexin-A Reverse Bone Mass Loss Induced by Chronic Intermittent Hypoxia Through OX1R-Nrf2/HIF-1α Pathway.

Background: Recent studies suggest that there is a potential connection between obstructive sleep apnea (OSA) and osteoporosis through dysregulation of bone metabolism. Orexin-A, a neuroprotective peptide secreted by the hypothalamus, is at a lower level in the plasma of OSA patients, which regulates appetite, energy expenditure and sleep-wake states. However, the protective effect of orexin-A on bone metabolism in OSA is unclear. Purpose: To investigate whether the activation of OX1R by orexin-A can reverse bone mass loss induced by chronic intermittent hypoxia (CIH). Methods: Mice were randomly divided into the normoxia group and CIH group. Within the CIH or normoxia groups, treatment groups were given a subcutaneous injection of either orexin-A or saline vehicle once every day for 4 weeks and then femurs were removed for micro-CT scans. Histology and immunohistochemical staining were performed to observe and calculate the changes in femurs as a result of hypoxia. Cell immunofluorescence and immunohistochemical staining were used to detect the expression of orexin receptors in MC3T3-E1 cells or in bones. CCK-8 assay, ALP assay kit and alizarin red staining were used to detect the viability, alkaline phosphatase (ALP) activity, and capacity of mineralization, respectively. The effect of orexin-A on osteogenic differentiation of MC3T3-E1 cells was evaluated using qRT-PCR, Western blot and cell staining. Results: CIH led to a decrease in the amount and density of trabecular bone, downregulated OCN expression while increasing osteoclast numbers in femurs and inhibited the expression of RUNX2, OSX, OPN and Nrf2 in MC3T3-E1 cells. Orexin-A treatment alleviated these CIH-induced effects by combining to OX1R. The level of HIF-1α was elevated both in CIH and orexin-A treatment groups. Conclusion: CIH environment inhibits osteogenesis and orexin-A can reverse bone mass loss induced by CIH through OX1R-Nrf2/HIF-1α pathway.

In situ mineralized PLGA/zwitterionic hydrogel composite scaffold enables high-efficiency rhBMP-2 release for critical-sized bone healing.

Osteoconductive and osteoinductive scaffolds are highly desirable for functional restoration of large bone defects. Here, we report an in situ mineralized poly(lactic-co-glycolic acid)/poly[2-(methacryloyloxy)ethyl]dimethyl-(3-sulfopropyl)ammonium hydroxide hydrogel (PLGA/PSBMA) scaffold as a novel high-efficiency carrier for recombinant human bone morphogenetic protein-2 (rhBMP-2) for bone tissue regeneration. By virtue of the oppositely charged structure, the zwitterionic PSBMA component is able to template well-integrated dense mineralization of calcium phosphate throughout the PLGA/PSBMA scaffold. The high affinity between rhBMP-2 and the mineralized matrix, combined with the capability of the zwitterionic hydrogel to sequester and to enable sustained release of ionic proteins, endows the mineralized PLGA/PSBMA scaffolds with high-efficiency sustained release of rhBMP-2 (only 1.7% release within 35 days), thus enabling robust healing of critical-sized (5 mm) nonunion calvarial defects in rats at an ultralow dosage of rhBMP-2 (150 ng per scaffold), at which level successful healing of critical-sized bone defects has never been reported. These findings show that the mineralized PLGA/PSBMA scaffold is promising for bone defect repair.

Comparison of diagnostic value of SWE, FNA and BRAF gene detection in ACR TI-RADS 4 and 5 thyroid nodules.

OBJECTIVES: To compare the diagnostic value of shear wave elastography (SWE), fine needle aspiration (FNA) and BRAF gene detection (BRAFV600E gene mutation detection) in ACR TI-RADS 4 and 5 thyroid nodules. METHODS: SWE images, FNA cytological results and BRAF detection results of ACR TI-RADS 4 and 5 thyroid nodules confirmed by pathology were analyzed retrospectively. The receiver operating characteristic (ROC) curve was drawn to determine the best cutoff value of SWE Emax. In the combined diagnosis of SWE, FNA and BRAF, firstly, the nodules with BRAF gene mutation were included in the positive ones, secondly, the nodules with benign and malignant FNA were included in the FNA + SWE or FNA + SWE + BRAF negative and positive ones respectively, finally, for FNA uncertain nodules: those whose SWE Emax were less than or equal to the cutoff value were included in FNA + SWE or FNA + SWE + BRAF negative ones, and those whose SWE Emax were greater than the cutoff value were included in positive ones. The diagnostic efficacy of SWE, FNA, SWE + FNA, FNA + BRAF and their combination in ACR TI-RADS 4 and 5 thyroid nodules were compared. RESULTS: The ROC curve showed that the best cutoff value of SWE Emax was 40.9 kpa, and the area under ROC curve (AUC) was 0.842 (0.800∼0.885). The sensitivity, specificity and accuracy of SWE were 76.3% (270/354), 75.5% (80/106) and 76.1% (350/460), respectively. The sensitivity, specificity and accuracy of FNA were 58.2% (206/354), 88.7% (94/106) and 65.2(300/460), respectively. The sensitivity, specificity and accuracy of FNA + BRAF were 95.5% (338/354), 88.7% (94/106) and 93.9% (432/460), respectively. The sensitivity, specificity and accuracy of SWE + FNA were 85.9% (304/354), 98.1% (104/106) and 88.7% (408/460), respectively. The sensitivity, specificity and accuracy of SWE + FNA + BRAF were 98.3% (348/354), 98.1% (104/106) and 98.3% (452/460), respectively. For the diagnostic accuracy, SWE + FNA + BRAF > FNA + BRAF > FNA + SWE > SWE > FNA, the difference was statistically significant (all P > 0.05). CONCLUSIONS: For ACR TI-RADS 4 and 5 thyroid nodules, SWE and FNA have high diagnostic efficiency. For the diagnostic accuracy, FNA + BRAF is better than FNA + SWE. FNA combination with BRAF gene detection further improves the diagnostic sensitivity and accuracy of FNA. The combined application of the three is the best.

Comparison of thyroid nodule FNA rates recommended by ACR TI-RADS, Kwak TI-RADS and ATA guidelines.

OBJECTIVES: To compare the differences among the thyroid imaging reporting and data system (TI-RADS) proposed by American College of Radiology (ACR TI-RADS), TI-RADS proposed by Kwak (Kwak TI-RADS), and American Thyroid Association (ATA) guidelines in the specificity, sensitivity and the unnecessary FNA rate (the UFR, the false positive rate) of recommended fine needle aspiration (FNA), and to observe the changes of the UFR in the KwakTI-RADS and ATA guidelines with the recommended FNA nodule size threshold. METHODS: The specificities, sensitivities and UFRs of recommended FNA in the ACR TI-RADS, ATA guidelines and Kwak TI-RADS were calculated and compared. The nodule sizes for recommended FNA of ATA guidelines and Kwak TI-RADS were systematically varied to establish new FNA thresholds. The specificities, sensitivities and UFRs of recommended FNA under the new models were calculated and compared to those in the ACR TI-RADS. RESULTS: For all thyroid nodules, the UFRs in the ACR TI-RADS, ATA guidelines and Kwak TI-RADS were 26.3%, 47.4% and 40.0%, respectively. The UFR in the ACR TI-RADS was lower than that in the others, and the specificity of recommended FNA in the ACR TI-RADS(73.7%) was higher than that in the others (all P < 0.001), but the sensitivity of recommended FNA in the ACR TI-RADS(89.3%) was lower than that in the others (all P < 0.001). When nodule sizes threshold of the recommended FNA for ATA guidelines Intermediate Suspicion, Low Suspicion, and Very Low Suspicion, and Kwak TI-RADS grade 4b and 4a were gradually increased, the UFRs gradually decreased. CONCLUSIONS: The UFRs of FNA recommended by the Kwak TI-RADS and ATA guidelines were higher than that of the ACR TI-RADS, and were affected by the recommended FNA nodule size threshold.

Bioinformatics analysis and experimental validation of differentially expressed genes in mouse articular chondrocytes treated with IL-1ß using microarray data.

Osteoarthritis (OA) is the most prevalent chronic degenerative disease that affects the health of the elderly. The present study aimed to identify significant genes involved in OA via bioinformatics analysis. A gene expression dataset (GSE104793) was downloaded from the Gene Expression Omnibus. Bioinformatics analysis was then performed in order to identify differentially expressed genes (DEGs) between untreated chondrocytes and chondrocytes cultured with interleukin-1ß (IL-1ß) for 24 h. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed using Metascape. A protein-protein interaction network of DEGs was constructed using the Search Tool for the Retrieval of Interacting Genes. Gene set enrichment analysis (GSEA) was performed using GSEA software. Furthermore, chondrocytes were extracted and treated with IL-1ß (10 ng/ml) for 24 h, and reverse-transcription quantitative PCR was used to confirm differential expression of hub genes. Patient samples were also collected to verify the bioinformatic analysis results. Based on the cut-off criteria used for determination of the DEGs, a total of 844 DEGs, including 498 upregulated and 346 downregulated DEGs, were identified. The DEGs were mainly enriched in the GO terms and KEGG pathways 'inflammatory response', 'negative regulation of cell proliferation', 'ossification', 'taxis', 'blood vessel morphogenesis', 'extracellular structure organization', 'mitotic cell cycle process' and 'TNF signaling pathway'. The majority of the PCR results, namely the differential expression of kininogen 2, complement C3, cyclin B1, cell division cycle 20, cyclin A2, 1-phosphatidylinositol 4-kinase, BUB1 mitotic checkpoint serine/threonine kinase, kinesin family member 11, cyclin B2 and BUB1 mitotic checkpoint serine/threonine kinase B were consistent with the bioinformatics results. Collectively, the present observations provided a regulation network of IL-1ß-stimulated chondrocytes, which may provide potential targets of OA therapy.

China experts' consensus on preventive and interceptive orthodontic treatments of malocclusions of children.

Malocclusion is one of the three most common oral diseases reported by World Health Organization(WHO). In China, its incidence rate is rising. Malocclusion seriously affects the dental and maxillofacial function, facial appearance and growth development of nearly 260 million children in China, and what is more, it affects their physical and mental health development. Malocclusion occurrence is related to genetic and environmental factors. Early treatment of malocclusion can create a good dental and maxillofacial development environment, correct abnormal growth and control the adverse effects of abnormal genetic factors. It can effectively reduce the prevalence of children's malocclusion and enhance their physical and mental health. This is an urgent need from the economic perspective of our society, so it has great practical and social significance. Experts from the project group "standard diagnose and treatment protocols for early orthodontic intervention of malocclusions of children" which initiated by China National Health Institute of Hospital Administration wrote the "China Experts' Consensus on Preventive and Interceptive Orthodontic Treatments of Malocclusions of Children", which aims to guide and popularize the clinical practice, improve the clinical theory and practice level, and accelerate the disciplinary development of early treatment of children's malocclusion in China. The consensus elaborates the harmfulness of malocclusion and the necessity of early treatment, and brings up the principles and fundamental contents. Based on the law of dental and maxillofacial development, this paper puts forward the guiding suggestions of preventive and interceptive treatments in different stages of dental development ranging from fetus to early permanent dentition. It is a systematic project to promote and standardize the early treatment of malocclusion. Through scientific and comprehensive stratified clinical practice and professional training, the clinical system of early treatment of malocclusion in China will eventually be perfected, so as to comprehensively care for children's dental and maxillofacial health, and improve their oral and physical health in China.

Smile Attractiveness Evaluation of Patients Selected for a U.S.-Based Board Certification Examination.

OBJECTIVES: The aims of this study were to assess smile attractiveness of a collection of 68 smiling photographs of successfully treated cases submitted to the American Board of Orthodontics (ABO) clinical examination and identify variables that influence the assessment. MATERIALS AND METHODS: A panel of 81 non-Caucasian assessors from various clinical disciplines were instructed to score the smile attractiveness on a visual analog scale from 1 (least attractive) to 10 (most attractive) and to select which components contributed to a lesser attractive smile. The mean, standard deviations (SDs), and quartiles of the smile attractiveness were obtained with descriptive statistics. Multilinear regression analysis was performed to investigate the scores of the perceived quality of smile attractiveness when the clinical disciplines and gender of the assessors were the factors taken into consideration. Receiver operating characteristic (ROC) curve was generated to establish the relationship between smile attractiveness and the achievement of a perfect smile. RESULTS: The mean (SD) rating of each clinical photograph of the anterior occlusion on smiling ranged from 3.11 (1.47) as the least attractive smile to 7.59 (1.45) as the most attractive smile. The overall mean (SD) score for smile attractiveness was 5.30 (1.10). Problems associated with teeth, gingiva, and lips corresponded with a reduction of the smile attractiveness score by 1.56, 1.82, and 1.47, respectively. Gender was not associated with smile attractiveness ratings. Orthodontists, periodontists, and prosthodontists demonstrated no difference in the ratings, while plastic surgeons were more critical than orthodontists regarding smile attractiveness. CONCLUSIONS: This study suggested that only 2 out of 68 AOB validated treatment finishes had a perfect and attractive smile.

Establishment of Detection Methods for Five Cannabinoids in Hemp Cosmetics Based on HPLC.

A simultaneously HPLC detection method for cannabidiolic acid (CBDA), cannabidiol (CBD), cannabinol (CBN), Δ9-tetrahydrocannabinol (THC), tetrahydro-cannabinolic acid (THCA) in 3 kinds of cosmetics matrix containing hemp leaf extract was developed. The extraction and HPLC conditions were optimized, and a methodological verification was also carried out. The results showed that this method had a good linear relationship in the range of 0.25 - 50 µg/mL with LOD values for 5 cannabinoids all between 0.10 - 0.25 µg/g. The recovery rates of 5 cannabinoids in 3 different cosmetics matrixes were between 90.1 - 108.5%, and the RSD values were all below 4.4%. These results indicated that this method had the advantages of simple operation, high sensitivity, and good accuracy. Through the testing of 6 kinds of hemp cosmetics, it was found that such cosmetics had uneven quality. The establishment of this method can lay a methodological foundation for establishing relevant testing method standards.

Effect of thickness and stacking order on Raman spectrum of layered CrCl3.

Layer- and stacking-dependent physical properties discovered in van der Waals materials have attracted a lot of attention recently. In this work, we have investigated the Raman spectrum of layered magnetic CrCl3with different atomic thickness and stacking order from first principles. Our study indicates Raman shift ofR3̄andC2/mphase is near degenerate except that the first and fourth Raman resonant modes exhibit a marked peak difference about 3 cm-1, which can be understood by stacking-dependent vibrational eigenvectors. Moreover, we also find that both frequency and intensity of Raman modes of few-layer CrCl3increase with the number of layers simultaneously. Furthermore, the Raman tensor and polarization-dependent Raman intensity are also evaluated in details, which are found to be in good agreement with available experiments. Our results proposed several distinctive signals to identify layer number and stacking order of layered CrCl3, which will provide an important reference to further experimental study.