Association between TRP channels and glutamatergic synapse gene polymorphisms and migraine and the comorbidities anxiety and depression in a Chinese population.

Background: Genetic and environmental factors contribute to migraine and the comorbidities of anxiety and depression. However, the association between genetic polymorphisms in the transient receptor potential (TRP) channels and glutamatergic synapse genes with the risk of migraine and the comorbidities of anxiety and depression remain unclear. Methods: 251 migraine patients containing 49 comorbidities with anxiety and 112 with depression and 600 controls were recruited. A customized 48-plex SNPscan kit was used for genotyping 13 SNPs of nine target genes. Logistic regression was conducted to analyze these SNPs' association with the susceptibility of migraine and comorbidities. The generalized multifactor dimension reduction (GMDR) was applied to analyze the SNP-SNP and gene-environment interactions. The GTEx database was used to examine the effects of the significant SNPs on gene expressions. Results: The TRPV1 rs8065080 and TRPV3 rs7217270 were associated with an increased risk of migraine in the dominant model [ORadj (95% CI): 1.75 (1.09-2.90), p = 0.025; 1.63 (1.02-2.58), p = 0.039, respectively]. GRIK2 rs2227283 was associated with migraine in the edge of significance [ORadj (95% CI) = 1.36 (0.99-1.89), p = 0.062]. In migraine patients, TRPV1 rs222741 was associated with both anxiety risk and depression risk in the recessive model [ORadj (95% CI): 2.64 (1.24-5.73), p = 0.012; 1.97 (1.02-3.85), p = 0.046, respectively]. TRPM8 rs7577262 was associated with anxiety (ORadj = 0.27, 95% CI = 0.10-0.76, p = 0.011). TRPV4 rs3742037, TRPM8 rs17862920 and SLC17A8 rs11110359 were associated with depression in dominant model [ORadj (95% CI): 2.03 (1.06-3.96), p = 0.035; 0.48 (0.23-0.96), p = 0.042; 0.42 (0.20-0.84), p = 0.016, respectively]. Significant eQTL and sQTL signals were observed for SNP rs8065080. Individuals with GRS (Genetic risk scores) of Q4 (14-17) had a higher risk of migraine and a lower risk of comorbidity anxiety than those with Genetic risk scores scores of Q1 (0-9) groups [ORadj (95% CI): 2.31 (1.39-3.86), p = 0.001; 0.28 (0.08-0.88), p = 0.034, respectively]. Conclusion: This study suggests that TRPV1 rs8065080, TRPV3 rs7217270, and GRIK2 rs2227283 polymorphism may associate with migraine risk. TRPV1 rs222741 and TRPM8 rs7577262 may associate with migraine comorbidity anxiety risk. rs222741, rs3742037, rs17862920, and rs11110359 may associate with migraine comorbidity depression risk. Higher GRS scores may increase migraine risk and decrease comorbidity anxiety risk.

Dual attention mechanism network for lung cancer images super-resolution.

BACKGROUND AND OBJECTIVE: Currently, the morbidity and mortality of lung cancer rank first among malignant tumors worldwide. Improving the resolution of thin-slice CT of the lung is particularly important for the early diagnosis of lung cancer screening. METHODS: Aiming at the problems of network training difficulty and low utilization of feature information caused by the deepening of network layers in super-resolution (SR) reconstruction technology, we propose the dual attention mechanism network for single image super-resolution (SISR). Firstly, the feature of a low-resolution image is extracted directly to retain the feature information. Secondly, several independent dual attention mechanism modules are constructed to extract high-frequency details. The introduction of residual connections can effectively solve the gradient disappearance caused by network deepening, and long and short skip connections can effectively enhance the data features. Furthermore, a hybrid loss function speeds up the network's convergence and improves image SR restoration ability. Finally, through the upsampling operation, the reconstructed high-resolution image is obtained. RESULTS: The results on the Set5 dataset for 4 × enlargement show that compared with traditional SR methods such as Bicubic, VDSR, and DRRN, the average PSNR/SSIM is increased by 3.33 dB / 0.079, 0.41 dB / 0.007 and 0.22 dB / 0.006 respectively. The experimental data fully show that DAMN can better restore the image contour features, obtain higher PSNR, SSIM, and better visual effect. CONCLUSION: Through the DAMN reconstruction method, the image quality can be improved without increasing radiation exposure and scanning time. Radiologists can enhance their confidence in diagnosing early lung cancer, provide a basis for clinical experts to choose treatment plans, formulate follow-up strategies, and benefit patients in the early stage.

Formation, Development, and Cross-Species Interactions in Biofilms.

Biofilms, which are essential vectors of bacterial survival, protect microbes from antibiotics and host immune attack and are one of the leading causes that maintain drug-resistant chronic infections. In nature, compared with monomicrobial biofilms, polymicrobial biofilms composed of multispecies bacteria predominate, which means that it is significant to explore the interactions between microorganisms from different kingdoms, species, and strains. Cross-microbial interactions exist during biofilm development, either synergistically or antagonistically. Although research into cross-species biofilms remains at an early stage, in this review, the important mechanisms that are involved in biofilm formation are delineated. Then, recent studies that investigated cross-species cooperation or synergy, competition or antagonism in biofilms, and various components that mediate those interactions will be elaborated. To determine approaches that minimize the harmful effects of biofilms, it is important to understand the interactions between microbial species. The knowledge gained from these investigations has the potential to guide studies into microbial sociality in natural settings and to help in the design of new medicines and therapies to treat bacterial infections.

Removal of the root canal smear layer using Carisolv III and sodium hypochlorite.

The present study investigated the effectiveness of a Carisolv III + 0.5% sodium hypochlorite (NaOCl)-based root canal irrigant for smear layer removal.Forty maxillary incisors were randomly divided into 4 groups (n = 10 per group). The canals in group A (experimental) were prepared with 0.5% NaOCl, and Carisolv III and 0.5% NaOCl was used for the final washing; groups B and C (positive controls) used 2% and 5.25% NaOCl, respectively; and group D (negative control) used phosphate-buffered saline (PBS). Ethylenediaminetetraacetic acid (EDTA) was used for all of the groups. A 5-point scoring scale and scanning electron microscopy were used to evaluate the effectiveness of the irrigants. The canals were consistently cleaner in the coronal and middle thirds than in the apical thirds (P < .05).For cleaning the root canals, 5.25% NaOCl was more effective than 2% NaOCl, 0.5% NaOCl + Carisolv III, and phosphate-buffered saline , respectively (P < .05). The 2% NaOCl solution showed similar results to 0.5% NaOCl + Carisolv III (P > .05). The combination of 5.25% NaOCl and 17% EDTA remains the most effective irrigant for removal of the root canal smear layer.A combination of Carisolv III + 0.5% NaOCl (with 17% EDTA) showed a cleaning ability similar to that of 2% NaOCl (with 17% EDTA).

Long noncoding RNA TUG1 promotes osteogenic differentiation of human periodontal ligament stem cell through sponging microRNA-222-3p to negatively regulate Smad2/7.

OBJECTIVE: To scrutinize the function of long non-coding RNA TUG1 on the osteogenic differentiation of human periodontal ligament stem cells in periodontitis and its specific mechanism. METHODS: Human periodontal ligament stem cells were extracted from human periodontium, followed by induction of osteogenic differentiation with osteogenic medium. Knockdown or overexpression of TUG1, microRNA-222-3p or Smad2/7 were performed in human periodontal ligament stem cells to observe their effect on expressions of osteogenic differentiation markers (Runx2, ALP, and OCN), and on calcium nodule formation by Alizarin red staining. Starbase software was utilized for prediction of the binding sites of TUG1 and microRNA-222-3p in addition to microRNA-222-3p and Smad2/7. Then dual-luciferase reporter gene assay was adopted to inspect the binding relationships between microRNA-222-3p and TUG1 or Smad2/7. RESULTS: Highly expressed TUG1 and lowly expressed microRNA-222-3p were found in human periodontal ligament stem cells during osteogenic differentiation. After measuring the expression of Runx2, ALP, and OCN as well as the formation of calcium nodules, we discovered that TUG1 can facilitate osteogenic differentiation of human periodontal ligament stem cells, while microRNA-222-3p had a reverse effect. Subsequently, knockdown of TUG1 and Smad2/7 or overexpression of microRNA-222-3p inhibited the osteogenic differentiation of human periodontal ligament stem cells. MicroRNA-222-3p is a target gene of TUG1, while microRNA-222-3p can negatively regulate Smad2/7. CONCLUSION: LncRNA TUG1 accelerates the osteogenic differentiation of human periodontal ligament stem cells by sponging microRNA-222-3p to regulate Smad2/7.

Root canal morphology of maxillary and mandibular first premolars analyzed using cone-beam computed tomography in a Shandong Chinese population.

The aim of this study was to explore the root morphology and root canal configuration of first premolars among Shandong Chinese residents using cone-beam computed tomography (CBCT).Randomly selected CBCT images were collected from 648 patients (44% women, 56% men). In total, 1268 maxillary and 1296 mandibular first premolars were analyzed. The number of roots and the canal configuration were recorded and identified based on Vertucci's classification.The majority of the maxillary first premolars had 1 root (67.4%), followed by 2 roots (32%). A 2-canal configuration (89%) was the most prevalent observation. For mandibular first premolars, 98.8% had 1 root and 81% presented the type I configuration. There were no statistical differences in the number of roots or morphology in terms of the left/right side or sex (P > .05).Among Chinese residents, the majority of maxillary first premolars had 1 root and 2 canals, whereas the most common anatomical configuration for mandibular first premolars was 1 root with 1 canal.

Design of stroke imaging package study of intracranial atherosclerosis: a multicenter, prospective, cohort study.

BACKGROUND: Intracranial atherosclerosis (ICAS) is a major cause of stroke worldwide. However, much remains unknown regarding its underlying pathophysiology. High-resolution magnetic resonance imaging (HR-MRI) can clearly display intracranial vessel wall lesions in vivo. The aim of stroke imaging package study of ICAS (SIPS-ICAS) study is to explore the stroke mechanisms of symptomatic ICAS, the dynamic changes under aggressive medical treatment and their associations with clinical events using conventional MRI sequences plus HR-MRI. METHODS: This is a multicenter, prospective, cohort study recruiting first-ever acute ischemic stroke patients attributed to intracranial large artery stenosis (>50% or occlusion). Subjects undergo a pre-designed stroke imaging packages at baseline and are recommended to receive aggressive medical treatments. Participants will be followed up for functional outcome, stroke recurrence, and death events at 3, 6 and 12 months and retake HR-MRI imaging at 6 months. RESULTS: Enrollment began in November 2018 and 96 patients have been enrolled as of September 2019. CONCLUSIONS: The SIPS-ICAS study will provide insights into the pathophysiology of ICAS and identify specific imaging markers for risk stratification and prognosis prediction. At the same time, the feasibility and validity of the new stroke imaging package including HR-MRI will be assessed, which is promising for clinical routine use.

Berberine Promotes Osteogenic Differentiation of Human Dental Pulp Stem Cells Through Activating EGFR-MAPK-Runx2 Pathways.

Similar to the mesenchymal stem cells (MSCs), dental pulp stem cells (DPSCs) also have pluripotent differentiation characteristic and may be more ideal for tissue regeneration, especially in tooth regeneration engineering. However, bacterial infection may be a powerful obstacle. Berberine (BBR), known with antibacterial effects, was recently found to play functions in bone formation through promoting osteogenic differentiation from pluripotent stem cells. However, whether BBR also function in DPSCs osteogenic differentiation has not yet been reported. Primary DPSCs were isolated from dental pulp tissues extracted from human impacted mandibular third molars, and identified by flow cytometry for cell surface antigen molecules. A dexamethasone osteogenic medium was used to induce DPSCs osteogenic differentiation. BBR (1 µM and 5 µM) was pre-added to into medium, and then cell proliferation, spheroid formation and osteogenic differentiation capacities of DPSCs were analyzed, as well as the underlying molecules modulation mechanism. Flow cytometry identified that CD44, CD90, CD81 and CD105 positively expressed in the isolated hDPSCs, with CD34 and CD45 negetively expressed. BBR enhanced the cell proliferation of hDPSCs in a dose-dependent pattern, and promoted dexamethasone-induced osteogenic differentiation via enhancing Runx2 transcription factor activity followed by upregulating osteogenesis markers expression, whereas the adipogenic differentiation of hDPSCs was suppressed dramatically by BBR. The EGFR and MAPK pathways were activated by BBR, and inhibitors for these pathways significantly suppressed the osteogenic differentiation promotion of BBR. These results have revealed a novel mechanism that berberine might promote hDPSCs osteogenic differentiation through activating EGFR-MAPK-Runx2 signaling pathways.

Berberine reduces inflammation of human dental pulp fibroblast via miR-21/KBTBD7 axis.

OBJECTIVE: This study is to investigate the effect and underlying mechanisms of berberine (BBR) on the proliferation and inflammatory levels of lipopolysaccharide induced human dental pulp fibroblast (LPS-HDPF). METHODS: Different concentrations of LPS were used to induce inflammatory response of HDPF. Cell proliferation was observed in BBR treated HDPF after transfection of miR-21 mimic, miR-21 inhibitor or sh-kBTBD7 plasmids and their negative controls. ELISA was employed to detect the expressions of inflammation related cytokines. Real-time quantitative RT-PCR was applied to estimate the expressions of miR-21 and KBTBD7. KBTBD7 expression was detected by Immunocytochemistry. Western blot analysis showed the protein levels of KBTBD7, Phospho-IKKα/ß, IKKß, Phospho-NF-κB p65, NF-κB p65, and IκBα. Luciferase reporter gene assay was used to determine the interaction between miR-21 and KBTBD7. RESULTS: LPS could promote inflammatory response in HDPF. Down-regulated miR-21 and up-regulated KBTBD7 were found in LPS-HDPF. BBR (25 uM) treatment in LPS-HDPF could ameliorate cell inflammatory response, presented by reduced expressions of IL-1ß, IL-6 and TNF-α, as well as enhanced cell proliferation and miR-21 expression. Moreover, miR-21 negatively targets KBTBD7. Over-expression of miR-21 or silencing of KBTBD7 could enhance the protective role of BBR on LPS-HDPF by inhibiting inflammatory response and promoting cell proliferation. Transfection of miR-21 overexpression or KBTBD7 silencing in BBR treated LPS-HDPF could inhibit activation of NF-κB signal pathway. CONCLUSION: Evidence in this study suggested that BBR mediates LPS induced inflammation in HDPF via miR-21/KBTBD7 axis to regulate NF-κB signal pathway, which may provide theoretical basis for BBR in prevention of pulpitis.

ADAM28 dramatically regulates the biological features of human gingival fibroblasts.

This study was to explore the effects of a disintegrin and metalloproteinase 28 (ADAM28) on the proliferation, differentiation, and apoptosis of human gingival fibroblasts (HGFs) and probable mechanism. After ADAM28 antisense oligodeoxynucleotide (AS-ODN) and sense oligodeoxynucleotide (S-ODN) were transfected into HGFs by Lipofectamine 2000, respectively, the expression discrepancies of ADAM28 among various groups were evaluated by reverse transcription-polymerase chain reaction (RT-PCR) and Western-blotting. Methabenzthiazuron (MTT) and cell-cycle assays were used to test the HGFs proliferation activity. Annexin V fluorescein isothiocyanate (FITC)/propidium iodide (PI) and alkaline phosphatase (ALP) analysis were performed separately to measure apoptosis and the cytodifferentiation standard. Immunocytochemistry and Western-blotting were carried out to determine the influence of ADAM28 AS-ODN on HGFs expressing core binding factor α1 (Cbfα1), cementum protein 1 (CEMP1), osteopontin (OPN) and dentin matrix protein 1 (DMP1). The AS-ODN group displayed the lowest expression level in HGFs, meanwhile the ADAM28 S-ODN group showed the highest. Furthermore, blocking of ADAM28 could inhibit the proliferation of HGFs, enhance HGFs differentiation and induce apoptosis of HGFs. Whereas, overexpression of ADAM28 generated the opposite effects and inhibited apoptosis. ADAM28 AS-ODN was able to notably suppress the expressions of Cbfα1 and CEMP1, and ADAM28 had positive correlations with cbfα1 and CEMP1. These provided conspicuous evidence that ADAM28 may play a crucial role in root development as a potential regulator of growth, differentiation, and apoptosis of HGFs.

[Maxillary first premolar with four canals: a case report].

This article presented a case of discovering and diagnosing three roots with four canals of the maxillary first premolar. We found and located the extra root canal by clinical diagnosis, careful observation during the operation, and multiangle X-ray. We further confirmed the existence of the three roots with four canals with the help of cone-beam computed tomography. Finally, we verified the success of the high-quality root-canal therapy through root optical microscopy.

miR-140-5p-mediated regulation of the proliferation and differentiation of human dental pulp stem cells occurs through the lipopolysaccharide/toll-like receptor 4 signaling pathway.

Human dental pulp stem cells (DPSCs) are oral mesenchymal stem cells with potential to differentiate into various cell types. Recent studies of DPSCs have focused on microRNAs (miRNAs), a class of small noncoding RNAs that play crucial roles in regulating DPSC phenotypes. In the current study, the expression of miR-140-5p was significantly decreased during lipopolysaccharide (LPS)-mediated differentiation of DPSCs in vitro. Overexpression of miR-140-5p enhanced proliferation of DPSCs and inhibited DPSC differentiation, whereas suppression of miR-140-5p produced the opposite effect. Moreover, the expression of toll-like receptor 4 (TLR-4), a critical regulator of DPSCs, was negatively correlated with the levels of miR-140-5p. A luciferase reporter analysis confirmed that miR-140-5p could regulate TLR-4 by directly binding to the 3'-untranslated region (3'-UTR) of the TLR4 mRNA. Additionally, we suppressed TLR-4 expression by treating cells with a TLR-4 inhibitor, CLI-095, and demonstrated that the effect of the miR-140-5p inhibitor on DPSC proliferation and differentiation could be partially reversed by blocking TLR-4. Taken together, our data suggest that miR-140-5p is a novel miRNA that regulates DPSC proliferation and differentiation.

Species discrimination in the subfamily Ostertagiinae of Northern China: assessment of DNA barcode in a taxonomically challenging group.

Gastrointestinal nematodes within the subfamily Ostertagiinae (Teladorsagia, Ostertagia, and Marshallagia et al.) are among the most common infections of domesticated livestock. These parasites are of particular interest, as many of the species within this group are of economic importance worldwide. Traditionally, nematode species designations have been based on morphological criteria. However, this group possesses poorly defined species. There is an urgent need to develop a reliable technique that can distinguish species of Ostertagiinae. DNA barcoding has been proved to be a powerful tool to identify species of birds, mammals, and arthropods, but this technique has not yet been examined for identifying species of Ostertagiinae. In this study, a total of 138 mitochondrial DNA (mtDNA) cytochrome c oxidase subunit I (COI) sequences from individuals representing 11 species of Ostertagiinae were acquired by PCR for the first time. The specimens were collected from pastoral area of northern China. Genetic divergence analyses showed that mean interspecific Kimura two-parameter distances of COI (13.61 %) were about four times higher than the mean value of the intraspecific divergence (3.69 %). Then, the performance of the COI to identify species of Ostertagiinae was evaluated by identification success rates using nearest neighbor (NN) and BLASTn. The results indicated that the rates of correct sequence identification for COI were high (>80 %) when using the NN and BLASTn methods. Besides, the deep lineage divergences are detected in Teladorsagia circumcincta. Meanwhile, the analyses also detected no genetic differentiation between some species such as Ostertagia hahurica and Ostertagia buriatica. These results indicate that the traditional status of species within Ostertagiinae should be closely examined based on the molecular data.