Role of the CTCF Binding Site in Human T-Cell Leukemia Virus-1 Pathogenesis.

During HTLV-1 infection, the virus integrates into the host cell genome as a provirus with a single CCCTC binding protein (CTCF) binding site (vCTCF-BS), which acts as an insulator between transcriptionally active and inactive regions. Previous studies have shown that the vCTCF-BS is important for maintenance of chromatin structure, regulation of viral expression, and DNA and histone methylation. Here, we show that the vCTCF-BS also regulates viral infection and pathogenesis in vivo in a humanized (Hu) mouse model of adult T-cell leukemia/lymphoma. Three cell lines were used to initiate infection of the Hu-mice, i) HTLV-1-WT which carries an intact HTLV-1 provirus genome, ii) HTLV-1-CTCF, which contains a provirus with a mutated vCTCF-BS which abolishes CTCF binding, and a stop codon immediate upstream of the mutated vCTCF-BS which deletes the last 23 amino acids of p12, and iii) HTLV-1-p12stop that contains the intact vCTCF-BS, but retains the same stop codon in p12 as in the HTLV-1-CTCF cell line. Hu-mice were infected with mitomycin treated or irradiated HTLV-1 producing cell lines. There was a delay in pathogenicity when Hu-mice were infected with the CTCF virus compared to mice infected with either p12 stop or WT virus. Proviral load (PVL), spleen weights, and CD4 T cell counts were significantly lower in HTLV-1-CTCF infected mice compared to HTLV-1-p12stop infected mice. Furthermore, we found a direct correlation between the PVL in peripheral blood and death of HTLV-1-CTCF infected mice. In cell lines, we found that the vCTCF-BS regulates Tax expression in a time-dependent manner. The scRNAseq analysis of splenocytes from infected mice suggests that the vCTCF-BS plays an important role in activation and expansion of T lymphocytes in vivo. Overall, these findings indicate that the vCTCF-BS regulates Tax expression, proviral load, and HTLV pathogenicity in vivo.

Novel bioactive nanospheres show effective antibacterial effect against multiple endodontic pathogens.

Aim: The current study evaluated the antibacterial activity of a newly developed quaternary ammonium polymethacrylate (QAPM)-containing bioactive glasses (BGs) via a two-step method by our group, namely BGs-HAEMB, and explored its cytotoxicity and biocompatibility. Methods: The antibacterial effects of the BGs-HAEMB against planktonic bacteria, bacterial biofilm formation, and experimental root canal biofilms of persistent pathogens (Enterococcus faecalis, Streptococcus sanguis and Porphyromonas endodontalis) associated with endodontic infection were evaluated in vitro by agar diffusion tests, direct contact tests and live/dead staining. The cytotoxicity and biocompatibility of BGs-HAEMB were evaluated by CCK-8 assays in vitro and a skin implantation model in vivo. Results: Compared to three clinically used endodontic sealers (Endofill, AH Plus, and iRoot SP), BGs-HAEMB exhibited the relatively strongest antibacterial effect against E. faecalis, S. sanguis and P. endodontalis after sitting for 14 and 28 days (P < 0.01). SEM images and CLSM images also showed that for each tested bacteria, BGs-HAEMB killed the most microorganism among all the experimental groups, regardless of treatment for 7 days or 28 days (P < 0.05). Besides, the BGs-HAEMB-treated groups showed a relatively low cytotoxicity (RGRs ranging from 88.6% to 102.9%) after 1, 3, and 7 days of exposure. Meanwhile, after 28 days of implantation, the inflammatory grade in BGs-HAEMB treated group was assessed as Grade I, in which the average numbers of inflammatory cells (6.7 ± 2.1) were less than 25. Conclusions: BGs-HAEMB exerted a long-term and stable antibacterial effect. The remarkable biocompatibility of BGs-HAEMB in vitro and in vivo confirmed its possible clinical application as a potential alternative in the development of the next generation of endodontic sealers.

An HTLV-1 envelope mRNA vaccine is immunogenic and protective in New Zealand rabbits.

Human T-cell leukemia virus type 1 (HTLV-1) is a retrovirus responsible for adult T-cell leukemia/lymphoma, a severe and fatal CD4+ T-cell malignancy. Additionally, HTLV-1 can lead to a chronic progressive neurodegenerative disease known as HTLV-1-associated myelopathy/tropical spastic paraparesis. Unfortunately, the prognosis for HTLV-1-related diseases is generally poor, and effective treatment options are limited. In this study, we designed and synthesized a codon optimized HTLV-1 envelope (Env) mRNA encapsulated in a lipid nanoparticle (LNP) and evaluated its efficacy as a vaccine candidate in an established rabbit model of HTLV-1 infection and persistence. Immunization regimens included a prime/boost protocol using Env mRNA-LNP or control green fluorescent protein (GFP) mRNA-LNP. After immunization, rabbits were challenged by intravenous injection with irradiated HTLV-1 producing cells. Three rabbits were partially protected and three rabbits were completely protected against HTLV-1 challenge. These rabbits were then rechallenged 15 weeks later, and two rabbits maintained sterilizing immunity. In Env mRNA-LNP immunized rabbits, proviral load and viral gene expression were significantly lower. After viral challenge in the Env mRNA-LNP vaccinated rabbits, an increase in both CD4+/IFN-γ+ and CD8+/IFN-γ+ T-cells was detected when stimulating with overlapping Env peptides. Env mRNA-LNP elicited a detectable anti-Env antibody response after prime/boost vaccination in all animals and significantly higher levels of neutralizing antibody activity. Neutralizing antibody activity was correlated with a reduction in proviral load. These findings hold promise for the development of preventive strategies and therapeutic interventions against HTLV-1 infection and its associated diseases.IMPORTANCEmRNA vaccine technology has proven to be a viable approach for effectively triggering immune responses that protect against or limit viral infections and disease. In our study, we synthesized a codon optimized human T-cell leukemia virus type 1 (HTLV-1) envelope (Env) mRNA that can be delivered in a lipid nanoparticle (LNP) vaccine approach. The HTLV-1 Env mRNA-LNP produced protective immune responses against viral challenge in a preclinical rabbit model. HTLV-1 is primarily transmitted through direct cell-to-cell contact, and the protection offered by mRNA vaccines in our rabbit model could have significant implications for optimizing the development of other viral vaccine candidates. This is particularly important in addressing the challenge of enhancing protection against infections that rely on cell-to-cell transmission.

The effect of Par3 on the cellular junctions and biological functions of odontoblast-lineage cells.

Perfect intercellular junctions are key for odontoblast barrier function. However, whether Partitioning defective-3 (Par3) is expressed in odontoblasts and its potential effects on odontoblast junctions are unknown. Herein, we investigated the effect of Par3 on cellular junctions and the biological behavior of odontoblast-lineage cells (OLCs). Whole-transcriptome sequencing was used to analyze the effects of Par3 on OLCs and the underlying molecular mechanism. Par3 was detected under physiological and inflammatory conditions in OLCs. To investigate the regulatory effect of Par3 on junctions between mouse OLCs, the effects of Par3 downregulation on the proliferation, migration, cycle and apoptosis of OLCs were detected by 5-ethyl-2'-deoxyuridine (EdU) and Transwell assays and flow cytometry. Western blotting and alizarin red S and alkaline phosphatase (ALP) staining were used to observe the effect of Par3 downregulation on OLC mineralization. Whole-transcriptome sequencing was used to investigate the biological role of Par3 in OLCs and potential molecular mechanisms. Par3 was located along the odontoblast layer in the rat pulp tissue and in the cytoplasm of OLCs. Par3 expression was downregulated under inflammatory conditions. The OLC junctions were discontinuous, and total Zona occluden-1 (ZO-1) expression and expression of ZO-1 at the membrane in OLCs were reduced after Par3 silencing (P < 0.05). Expression of a junction-related protein (ZO-1) was downregulated after the downregulation of Par3 (P < 0.05), and ZO-1 moved from the cell membrane to the cytoplasm. OLC proliferation and migration were enhanced, but apoptosis and mineralization were inhibited in shPar3-transfected cells (P < 0.05). Sequencing identified 2996 differentially expressed genes (DEGs), which were mainly enriched in the response to stimuli and binding. Downregulation of Par3 could overactivate the PI3k-AKT pathway by promoting AKT phosphorylation (P < 0.05). Downregulation of Par3 may disrupt junctions between OLCs by affecting ZO-1 expression and distribution and promote OLC proliferation and migration but inhibit OLC mineralization. Par3 may interact with 14-3-3 proteins for PI3K-AKT pathway activation to affect OLC junctions and function.

Synthesis and biological evaluation of Ribo 7-N/O/S pyrimidine 9-deaza C-nucleoside analogs as new antiviral agents for inhibiting HCV RNA-dependent RNA polymerases.

Hepatitis C infection is caused by the bloodborne pathogen hepatitis C virus (HCV) and can lead to serious liver diseases and, ultimately, death if the treatment is ineffective. This work reports the synthesis and preclinical evaluation of 7 novel 9-O/N/S pyrimidine nucleosides, including compound 12, the triphosphate of known compound 7b. The nucleosides are 9-deaza modifications of adenosine and guanosine with ß-2'-C-methyl substituent on the ribose. Within this series of compounds, a 9-deaza furopyrimidine analog of adenosine, compound 7b, showed high anti-HCV activity in vitro, good stability, low toxicity, and low genotoxicity when administrated in low doses, and an adequate pharmacokinetics profile. An improved synthesis of compound 7b compared to a previous study is also reported. Compound 12 was synthesized as a control to verify phosphorylation of 7b occurred in vivo.

Computer Vision Technology for Monitoring of Indoor and Outdoor Environments and HVAC Equipment: A Review.

Artificial intelligence technologies such as computer vision (CV), machine learning, Internet of Things (IoT), and robotics have advanced rapidly in recent years. The new technologies provide non-contact measurements in three areas: indoor environmental monitoring, outdoor environ-mental monitoring, and equipment monitoring. This paper summarizes the specific applications of non-contact measurement based on infrared images and visible images in the areas of personnel skin temperature, position posture, the urban physical environment, building construction safety, and equipment operation status. At the same time, the challenges and opportunities associated with the application of CV technology are anticipated.

Effect of full pulpotomy using a calcium silicate-based bioactive ceramic in adult permanent teeth with symptoms indicative of irreversible pulpitis: A retrospective study.

BACKGROUND: The authors studied the treatment effect of full pulpotomy using a calcium silicate-based bioactive ceramic in adult permanent teeth with symptoms indicative of irreversible pulpitis. METHODS: Eighty-one adult permanent teeth with symptoms indicative of irreversible pulpitis in 78 patients aged 18 through 72 years were evaluated for inclusion in the study. After caries excavation, the pulp was amputated to the level of the canal orifices. After hemostasis was achieved, calcium silicate-based bioactive ceramic was placed as the capping agent. The cavity was sealed temporarily with a glass ionomer cement and then restored with flowable resin and composite resin after 2 weeks if no positive symptoms were reported or detected. Postoperative evaluation was performed by means of clinical and radiographic examination at 2 weeks and 3, 6, and 12 months. RESULTS: Overall success rates of the procedure were 96.3% (78 of 81), 93.8% (76 of 81), 92.6% (75 of 81), and 92.6% (75 of 81) at the 2-week, 3-month, 6-month, and 12-month recall visits, respectively. Six of the 81 teeth failed and required root canal therapy. In these 6 teeth, 3 exhibited severe cold stimuli pain and spontaneous pain at the 2-week follow-up, 2 had no response to electric pulp testing with apical percussion pain and periapical rarefaction at the 3-month follow-up, and 1 tooth exhibited periapical rarefaction and labial mucosal fistula at the 6-month follow-up. CONCLUSIONS: Under the conditions of this study, full pulpotomy using a calcium silicate-based bioactive ceramic was a successful option for the treatment of adult permanent teeth with carious originated symptoms indicative of irreversible pulpitis. PRACTICAL IMPLICATIONS: Vital pulp therapy is no longer impossible for adult permanent teeth with carious originated symptoms indicative of irreversible pulpitis.

Effect of access cavities on the biomechanics of mandibular molars: a finite element analysis.

INTRODUCTION: This study aimed to predict the fracture resistance of a mandibular first molar (MFM) with diverse endodontic cavities using finite element analysis (FEA). METHODS: Five experimental finite element models representing a natural tooth (NT) and 4 endodontically treated MFMs were generated. Treated MFM models were with a traditional endodontic cavity (TEC) and minimally invasive endodontic (MIE) cavities, including guided endodontic cavity (GEC), contracted endodontic cavity (CEC) and truss endodontic cavity (TREC). Three loads were applied, simulating a maximum bite force of 600 N (N) vertically and a normal masticatory force of 225 N vertically and laterally. The distributions of von Mises (VM) stress and maximum VM stress were calculated. RESULTS: The maximum VM stresses of the NT model were the lowest under normal masticatory forces. In endodontically treated models, the distribution of VM stress in GEC model was the most similar to NT model. The maximum VM stresses of the GEC and CEC models under different forces were lower than those of TREC and TEC models. Under vertical loads, the maximum VM stresses of the TREC model were the highest, while under the lateral load, the maximum VM stress of the TEC model was the highest. CONCLUSION: The stress distribution of tooth with GEC was most like NT. Compared with TECs, GECs and CECs may better maintain fracture resistance, TRECs, however, may have a limited effect on maintenance of the tooth resistance.

Dysbiosis of Oral Microbiota and Metabolite Profiles Associated with Type 2 Diabetes Mellitus.

Several previous studies have shown that oral microbial disorders may be closely related to the occurrence and development of type 2 diabetes mellitus (T2DM). However, whether the function of oral microorganisms and their metabolites have changed in patients with T2DM who have not suffered from any oral diseases has not been reported. We performed metagenomic analyses and nontargeted metabolic analysis of saliva and supragingival plaque samples from patients with T2DM who have not suffered any oral diseases and normal controls. We found that periodontal pathogens such as Porphyromonas gingivalis and Prevotella melaninogenica were significantly enriched, while the abundances of dental caries pathogens such as Streptococcus mutans and Streptococcus sobrinus were not significantly different in patients with T2DM compared to those in normal controls. Metabolomic analyses showed that the salivary levels of cadaverine and L-(+)-leucine of patients with T2DM were significantly higher than those of normal controls, while the supragingival plaque levels of N-acetyldopamine and 3,4-dimethylbenzoic acid in patients with T2DM were significantly higher than those in the normal controls. Additionally, we identified the types of oral microorganisms related to the changes in the levels of circulating metabolites, and the oral microorganisms were involved in the dysregulation of harmful metabolites such as cadaverine and n, n-dimethylarginine. Overall, our study first described the changes in the composition of oral microorganisms and their metabolites in patients with T2DM who have not suffered any oral diseases, which will provide a direct basis for finding oral biomarkers for early warning of oral diseases in T2DM. IMPORTANCE The incidence of oral diseases in type 2 diabetic patients might increase, and the severity might also be more serious. At present, the relationship between oral microorganisms and type 2 diabetes mellitus (T2DM) has become a hot topic in systemic health research. However, whether the function of oral microorganisms and their metabolites have changed in patients with T2DM who have not suffered from any oral diseases has not been reported. We found that even if the oral condition of T2DM is healthy, their oral microbes and metabolites have changed, thus increasing the risk of periodontal disease. Our study first described the changes in the composition of oral microorganisms and their metabolites in T2DM who have not suffered any oral diseases and revealed the correlation between oral microorganisms and their metabolites, which will provide a direct basis for finding oral biomarkers for early warning of oral diseases in patients with T2DM.

Transforming growth factor-ß1 promotes early odontoblastic differentiation of dental pulp stem cells via activating AKT, Erk1/2 and p38 MAPK pathways.

Background/purpose: TGF-ß1 (Transforming growth factor-ß1) plays an important role in the regeneration and repair of pulp-dentin complex. However, the biological function of TGF-ß1 on odontoblastic differentiation remains unclear, mainly due to the processes of differentiation were controlled by complex signaling pathways. This study aimed to investigate the signaling pathways involved in regulating the early differentiation of dental pulp stem cells (DPSCs) by TGF-ß1 and their functional role. Materials and methods: DPSCs were treated with 1 ng/mL TGF-ß1 and Western blotting was conducted to examine the activation of protein kinase B (AKT), small mothers against decapentaplegic 3 (Smad3), p38 mitogen-activated protein kinase (p38 MAPK), c-Jun N-terminal kinase (JNK) and extracellular signal-regulated kinase 1/2 (Erk1/2). DPSCs were exposed to mineralization medium contained TGF-ß1 with/without the specific signaling pathway inhibitors, and early odontogenic differentiation was evaluated by assessing the expression of alkaline phosphatase (ALP), collagen type 1 alpha 1 (COL1A), dentin matrix protein 1 (DMP-1) and runt-related transcription factor 2 (Runx2). Results: TGF-ß1 stimulated AKT, Smad3, p38 MAPK, Erk1/2 and JNK phosphorylation in DPSCs within 120 min. TGF-ß1 enhanced ALP activity and elevated levels of COL1A, DMP-1 and Runx2. LY294002, U0126 and SB203580 attenuated the effect of TGF-ß1 on DPSCs, however, the SIS3 and SP600125 treated groups had no significant effect. Conclusion: TGF-ß1 promotes the early stage of odontoblastic differentiation in DPSCs by activating AKT, Erk1/2 and p38 MAPK signaling pathways, but not by Smad3 and JNK.

Synthesis of novel azasugar-containing 2'ß-C-Me 9-deaza nucleosides as potential anti-hepatitis C virus agents.

As a part of our ongoing discovery efforts exploring azasugar as agents for treating various unmet medical needs, we prepared analogs of azasugar as potential anti-hepatitis C virus (HCV) agents. Herein we describe the synthesis of novel 2'ß-C-Me 9-deazanucleoside azasugar analogs.

Research progress on root canal irrigation disinfection drugs / 口腔疾病防治

@#Endodontic infection control is crucial to successful root canal treatment. Irrigation is the key step in endodontic procedures, and the application of root canal irrigation and disinfection medications play an important role. How to enhance antibacterial effects and functions in removing tissues while maintaining biocompatibility is a hot topic in endodontics. Currently, insights to address this issue can be split into two categories: one, the modification or combination of conventional endodontic irrigation solutions, and two, the development of novel endodontic irrigation solutions with new technologies and materials, for instance, nanomaterials and natural exacts. However, conventional endodontic irrigation solutions, such as sodium hypochlorite and chlorhexidine, are still the first choice in clinical practice. Most novel endodontic irrigation solutions remain at the pre-clinical laboratory stage. Clinical research and relevant data are required to determine whether various methods can improve endodontic irrigation. From basic research to clinical application is the direction for advancing to the next stage. The present article focuses on research progress on endodontic irrigation, especially concerning its antibacterial mechanism, characteristics and efficacy, to provide a reference for future clinical translation.

Contactless sleep posture measurements for demand-controlled sleep thermal comfort: A pilot study.

Thermal comfort during sleep is essential for both sleep quality and human health while sleeping. There are currently few effective contactless methods for detecting the sleep thermal comfort at any time of day or night. In this paper, a vision-based detection approach for human thermal comfort while sleeping was proposed, which is intended to avoid overcooling/overheating supply, meet the thermal comfort needs of human sleep, and improve human sleep quality and health. Based on 438 valid questionnaire surveys, 10 types of thermal comfort sleep postures were summarized. By using a large number of data captured, a fundamental framework of detection algorithm was constructed to detect human sleeping postures, and corresponding weighting model was established. A total of 2.65 million frames of posture data in natural sleep status were collected, and thermal comfort-related sleep postures dataset was created. Finally, the robustness and effectiveness of the proposed algorithm were validated. The validation results show that the sleeping posture and human skeleton keypoints can be used for estimating sleeping thermal comfort, and the the quilt coverage area can be fused to improve the detection accuracy.

Scaffold hopping via ring opening enables identification of acyclic compounds as new complement Factor D inhibitors.

The three complement pathways comprising the early phase of the complement system (the classical, lectin, and alternative pathways) act together with the innate and adaptive immune systems to protect against foreign entities and maintain tissue homeostasis. While these systems are normally under tight regulatory control, several diseases have been reported to correlate with uncontrolled activation and amplification of the alternative pathway, including paroxysmal nocturnal hemoglobinuria, atypical hemolytic uremic syndrome, C3 glomerulopathy, and age-related macular degeneration. Complement FactorD (CFD), a serine protease, is the rate-limiting enzyme for the activity of alternative pathway. CFD activates the alternative pathway by cleaving Complement Factor B complexed to C3b (C3bB) to generate alternative pathway C3 convertase (C3bBb). In our search for novel CFD inhibitors with therapeutic potential, we employed a hot-spot analysis of an ensemble of apo and holo CFD structures. This analysis identified potential pharmacophore features that aided in the design of a series of compounds based on an l-proline core. While these compounds inhibited CFD in an esterolytic assay (for example, a proline-based compound, IC50 = 161 nM), the pharmacokinetic (PK) properties were poor. A strategy of scaffold hopping via ring opening led to a novel series of acyclic compounds, with subsequent structure-based ligand design and lead optimization producing several novel CFD inhibitors. One of these inhibitors, 1-(2-((2-(3-chloro-2-fluorobenzylamino)-2-oxoethyl)(cyclopropyl)amino)-2-oxoethyl)-5-(3-methyl-3-(1-methylpiperidin-4-yl)ureido)-1H-indazole-3-carboxamide, showed good potency with IC50s of 37 nM in the esterolytic assay and 30 nM in a hemolytic assay and PK assessments following oral administration to rats revealed a Cmax of 113 ng/mL and an AUC0-24h of 257 hr.ng/mL.

Endodontic management of a fused left maxillary second molar and two paramolars using cone beam computed tomography: A case report.

BACKGROUND: Fused teeth usually involve several complications, such as the development of caries in the groove between fused crowns, tooth impaction, diastemas, aesthetic and periodontal problems, and pulpal pathosis, due to the complex anatomical structure of fused teeth. A thorough diagnosis is paramount to forming an accurate treatment plan and obtaining a favourable prognosis. With the advent of cone-beam computed tomography (CBCT), accurate 3-dimensional images of teeth and their surrounding dentoalveolar structures can now be readily obtained, and the technology can accurately provide a minimally invasive approach to acquire detailed diagnostic information. Therefore, we utilize CBCT data herein to generate a digital model for the infected region in a patient, and this model enables us to better plan the management of his case. CASE SUMMARY: This report details the diagnosis and endodontic treatment of a rare case involving a fused maxillary second molar and two paramolars with apical periodontitis. The patient experienced pain upon biting and cold sensitivity in the area of the maxillary left molar. No caries or other defects were identified in these teeth, and a normal response to a pulp electric viability test was observed. With the aid of CBCT and digital model technology, we initially suspected that the infection originated from the isthmus between the maxillary second molar and two paramolars. Therefore, we only treated the isthmus by an endodontic approach and did not destroy the original tooth structure; furthermore, the vital pulp was retained, and good treatment outcomes were observed at the 24-month follow-up. CONCLUSION: This finding may provide new insights and perspectives on the diagnosis and treatment of fused teeth.

Enamel Matrix Derivative Enhances the Odontoblastic Differentiation of Dental Pulp Stem Cells via Activating MAPK Signaling Pathways.

The odontoblastic differentiation of dental pulp stem cells (DPSCs) contributes to pulp-dentin regeneration. Enamel matrix derivative (EMD) is considered to be a critical epithelial signal to induce cell differentiation during odontogenesis and has been widely applied to clinical periodontal tissue regeneration. The purpose of this study was to explore the effect of EMD on DPSCs proliferation and odontoblastic differentiation, as well as the underlying mechanisms. We conducted in vitro and in vivo researches to get a comprehensive understanding of EMD. In vitro phase: cell proliferation was assessed by a cell counting kit-8 (CCK-8) assay; then, alkaline phosphatase (ALP) activity and staining, alizarin red staining, real-time RT-PCR, and western blot analysis were conducted to determine the odontoblastic potential and involvement of MAPK signaling pathways. In vivo phase: after ensuring the biocompatibility of VitroGel 3D-RGD via scanning electron microscopy (SEM), the hydrogel mixture was subcutaneously injected into nude mice followed by histological and immunohistochemical analyses. The results revealed that EMD did not interfere with DPSCs proliferation but promoted the odontoblastic differentiation of DPSCs in vitro and in vivo. Furthermore, blocking the MAPK pathways suppressed the EMD-enhanced differentiation of DPSCs. Finally, VitroGel 3D-RGD could well support the proliferation, differentiation, and regeneration of DPSCs. Overall, this study demonstrates that EMD enhances the odontoblastic differentiation of DPSCs through triggering MAPK signaling pathways. The findings provide a new insight into the mechanism by which EMD affects DPSCs differentiation and proposes EMD as a promising candidate for future stem cell therapy in endodontics.

Cubic Nanogrids for Counterbalance Contradiction among Reorganization Energy, Strain Energy, and Wide Bandgap.

Molecular cross-scale gridization and polygridization of organic π-backbones make it possible to install 0/1/2/3-dimensional organic wide-bandgap semiconductors (OWBGSs) with potentially ZnO-like fascinating multifunctionality such as optoelectronic and piezoelectronic features. However, gridization effects are limited to uncover, because the establishment of gridochemistry still requires a long time, which offers a chance to understand the effects with a theoretical method, together with data statistics and machine learning. Herein, we demonstrate a state-of-the-art 3D cubic nanogridon with a size of ∼2 × 2 × 1.5 nm3 to examine its multigridization of π-segments on the bandgap, molecular strain energy (MSE), as well as reorganization energy (ROE). A cubic gridon (CG) consists of a four-armed bifluorene skeleton and a thiophene-containing fused arene plane with the Csp3 spiro-linkage, which can be deinstalled into face-on or edge-on monogrids. As a result, multigridization does not significantly reduce bandgaps (Eg ≥ 4.03 eV), while the MSE increases gradually from 4.72 to 23.83 kcal/mol. Very importantly, the ROE of a CG exhibits an extreme reduction down to ∼28 meV (λ+) that is near the thermal fluctuation energy (∼26 meV). Our multigridization results break through the limitation of the basic positively proportional relationship between reorganization energies and bandgaps in organic semiconductors. Furthermore, multigridization makes it possible to keep the ROE small under the condition of a high MSE in OWBGS that will guide the cross-scale design of multifunctional OWBGSs with both inorganics' optoelectronic performance and organics' mechanical flexibility.

Profiling long noncoding RNA alterations during the stromal cell-derived factor-1α-induced odontogenic differentiation of human dental pulp stem cells.

OBJECTIVE: The purpose of this study was to investigate the differential expression of long noncoding RNAs (lncRNAs) in dental pulp stem cells (DPSCs) after stromal cell-derived factor-1α (SDF-1α) induction and to explore the lncRNAs that regulate the odontogenic differentiation and migration of DPSCs. DESIGN: We examined the altered expression of lncRNAs in DPSCs after SDF-1α induction by performing lncRNA microarray and qRT-PCR analyses. Moreover, a bioinformatics analysis was conducted to predict the interactions of lncRNAs and identify core regulatory factors. A small interfering RNA (siRNA) was used to knock down lncRNA AC080037.1 expression in DPSCs. Cell transmigration assays, alizarin red staining, qRT-PCR and Western blotting were performed to detect the expression of osteo/dentinogenic differentiation markers or Rho GTPase after lncRNA knockdown in DPSCs. RESULTS: The microarray analysis identified 206 differentially expressed lncRNAs at 7 days after treatment. One lncRNA, AC080037.1, was shown to regulate the odontogenic differentiation of DPSCs. An siRNA targeting lncRNA AC080037.1 suppressed DPSCs migration and the expression of Rho GTPase induced by SDF-1α. Moreover, AC080037.1 knockdown significantly affected mineralized nodule formation and substantially suppressed runt-related factor-2 (RUNX-2), dentin matrix protein-1 (DMP-1) and dentin sialophosphoprotein (DSPP) expression in DPSCs. CONCLUSIONS: Our results revealed the differential expression of lncRNAs in DPSCs before and after SDF-1α induction. Furthermore, we highlighted the significant involvement of one lncRNA, AC080037.1, in the positive regulation of the osteo/odontogenic differentiation of DPSCs and indicated that this lncRNA might be a potential target in regenerative endodontics. These findings may further advance translational studies of pulp engineering.

Heparanase Blockade as a Novel Dual-Targeting Therapy for COVID-19.

The coronavirus disease 2019 (COVID-19) pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has caused over 5 million deaths worldwide. Pneumonia and systemic inflammation contribute to its high mortality. Many viruses use heparan sulfate proteoglycans as coreceptors for viral entry, and heparanase (HPSE) is a known regulator of both viral entry and inflammatory cytokines. We evaluated the heparanase inhibitor Roneparstat, a modified heparin with minimum anticoagulant activity, in pathophysiology and therapy for COVID-19. We found that Roneparstat significantly decreased the infectivity of SARS-CoV-2, SARS-CoV-1, and retroviruses (human T-lymphotropic virus 1 [HTLV-1] and HIV-1) in vitro. Single-cell RNA sequencing (scRNA-seq) analysis of cells from the bronchoalveolar lavage fluid of COVID-19 patients revealed a marked increase in HPSE gene expression in CD68+ macrophages compared to healthy controls. Elevated levels of HPSE expression in macrophages correlated with the severity of COVID-19 and the expression of inflammatory cytokine genes, including IL6, TNF, IL1B, and CCL2. In line with this finding, we found a marked induction of HPSE and numerous inflammatory cytokines in human macrophages challenged with SARS-CoV-2 S1 protein. Treatment with Roneparstat significantly attenuated SARS-CoV-2 S1 protein-mediated inflammatory cytokine release from human macrophages, through disruption of NF-κB signaling. HPSE knockdown in a macrophage cell line also showed diminished inflammatory cytokine production during S1 protein challenge. Taken together, this study provides a proof of concept that heparanase is a target for SARS-CoV-2-mediated pathogenesis and that Roneparstat may serve as a dual-targeted therapy to reduce viral infection and inflammation in COVID-19. IMPORTANCE The complex pathogenesis of COVID-19 consists of two major pathological phases: an initial infection phase elicited by SARS-CoV-2 entry and replication and an inflammation phase that could lead to tissue damage, which can evolve into acute respiratory failure or even death. While the development and deployment of vaccines are ongoing, effective therapy for COVID-19 is still urgently needed. In this study, we explored HPSE blockade with Roneparstat, a phase I clinically tested HPSE inhibitor, in the context of COVID-19 pathogenesis. Treatment with Roneparstat showed wide-spectrum anti-infection activities against SARS-CoV-2, HTLV-1, and HIV-1 in vitro. In addition, HPSE blockade with Roneparstat significantly attenuated SARS-CoV-2 S1 protein-induced inflammatory cytokine release from human macrophages through disruption of NF-κB signaling. Together, this study provides a proof of principle for the use of Roneparstat as a dual-targeting therapy for COVID-19 to decrease viral infection and dampen the proinflammatory immune response mediated by macrophages.

Metagenomic study on the composition and function of oral microorganisms in healthy adults / 口腔疾病防治

Objective@#Metagenomic sequencing was used to explore the species composition and internal functional metabolic pathway of saliva and supragingival plaque microbial communities in healthy adults to provide a theoretical reference for the biological prevention and treatment of oral diseases.@*Methods@#Saliva and supragingival plaque samples were collected from healthy adults, total DNA was extracted, and a metagenomic library was constructed. The qualified library was sequenced via metagenomics, and the sequencing data were analyzed using bioinformatics and statistics. @*Results @#The main bacterial phyla in healthy oral samples were Proteobacteria (32.51%), Bacteroidetes (30.81%), and Actinobacteria (16.23%), and the main bacterial species were Corynebacterium matruchotii (3.84%), Haemophilus parainfluenzae (2.91%), and Prevotella melaninogenica (2.76%). The alpha diversity of the supragingival plaque group was higher than that of the saliva group, and there was a significant difference in the composition of the microbial community between the two groups (P<0.05). At the species level, Prevotella melaninogenica, Fusobacterium periodonticum, and Prevotella intermedia were more abundant in saliva samples than in supragingival plaque samples, while Corynebacterium matruchotii, Propionibacterium acidifaciens, and Rothia dentocariosa were more abundant in supragingival plaque samples than in saliva samples (P<0.05). High-quality gene sets of saliva and supragingival plaque in healthy adults were constructed based on metagenomic sequencing. The results of KEGG pathway functional metabolic differences showed that starch and sucrose metabolism, leucine and isoleucine degradation, and arginine biosynthesis in salivary microorganisms were more abundant than in supragingival plaque, while glycolysis/gluconeogenesis and carbon metabolism in supragingival plaque were more abundant than in saliva.@* Conclusion@#There are significant differences in the species composition and functional gene metabolic pathways of saliva and supragingival plaque microecology in healthy adults. The sensitivity of dominant species in different microecological regions to the identification of oral diseases may be different. In the microbiological study of oral diseases, appropriate samples should be selected according to different diseases.