Clinical and hematological characteristics of children infected with the omicron variant of SARS-CoV-2: role of the combination of the neutrophil: lymphocyte ratio and eosinophil count in distinguishing severe COVID-19.

Purpose: Investigate the clinical/hematological characteristics of children infected with the Omicron variant of severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) and identify an effective indicator to distinguish coronavirus disease 2019 (COVID-19) severity in children. Methods: A retrospective study was conducted through electronic medical records from pediatric patients. The demographic, clinical, and routine blood test (RBT) features of children diagnosed by real-time PCR for SARS-CoV-2 were collected. Results: Data of 261 patients were analyzed. The most common abnormality shown by RBTs was increased monocyte count (68%). Children had "mild-moderate" or "severe" forms of COVID-19. Prevalence of abnormal neutrophil count (p = 0.048), eosinophil count (p = 0.006), mean corpuscular volume (p = 0.033), mean platelet volume (p = 0.006), platelet-large cell ratio (p = 0.043), and red blood cell distribution width-standard deviation (p = 0.031) were significantly different in the two types. A combination of the neutrophil: lymphocyte ratio (NLR) and eosinophil count for diagnosing severe COVID-19 presented the largest AUC (0.688, 95% CI = 0.599-0.777; p < 0.001), and the AUC increased with a decrease in age. Conclusions: Combination of the NLR and eosinophil count might be a promising indicator for identifying severe COVID-19 in children at infection onset.

Preparation of MXene/BN Composites with Adjustable Microwave Absorption Performance.

The challenge of developing a high-efficiency microwave absorbent remains, because of the compatibility between microwave absorption and high-temperature-resistant performance in practical application. Herein, a facile method is used to obtain serial MXene/BN-zxy composites, where zx:y indicates the weight ratio of MXene and boron nitride (BN) in the composites, with adjustable microwave absorption performance (MAP) which can be regulated by the ratio of MXene and the BN nanosheet. In particular, the as-prepared absorbents with supercapacitance-like structure significantly enhanced the MAP and could be served more than 900 °C. The results of MAP reveal that the minimum reflection loss (RL) can reach -20.94 dB with a MXene/BN-101 composite coating thickness of 4.0 mm; the effective attenuation bandwidth (RL< -10 dB, i.e., 90% microwave energy is attenuated) is up to 9.71 GHz (7.94-17.65 GHz). From a detailed analysis, it is observed that attenuation is the critical limiting factor for MAPs rather than impedance mismatch, which can be assigned to the poor MAP of BN nanosheets. In any case, as-prepared absorbents have potential applications in the field of heating components.

Effect of polyhydroxy-terminated PAMAM dendrimer on dentin matrix metalloproteinases within the hybrid layers.

BACKGROUND: Intrafibrillar remineralization within the hybrid layers (HLs) has recently attracted extensive attention in achieving durable resin-dentin bonds. The polyhydroxy-terminated poly(amidoamine) dendrimer (PAMAM-OH) at fourth generation becomes a desirable candidate to induce intrafibrillar remineralization to protect exposed collagen fibrils within HLs based on the size exclusion effect of fibrillar collagen. However, the remineralization process in vivo is time-consuming, during which the exposed collagen fibrils are vulnerable to enzymatic degradation, resulting in unsatisfactory remineralization. Thereby, if PAMAM-OH itself possesses concomitant anti-proteolytic activity during the induction of remineralization, it would be very beneficial to obtain satisfactory remineralization. METHODS: Binding capacity tests using adsorption isotherm and confocal laser scanning microscopy (CLSM) were performed to assess if the PAMAM-OH had adsorption capacity on dentin. Anti-proteolytic testings were detected by MMPs assay kit, in-situ zymography and ICTP assay. Adhesive infiltration of resin-dentin interface and tensile bond strength before and after thermomechanical cycling were implemented to assess if the PAMAM-OH adversely affected resin-dentin bonds. RESULTS: Anti-proteolytic testings performed using MMPs assay kit, in-situ zymography and ICTP assay indicated that PAMAM-OH inhibited exogenous soluble MMP-9 as well as had inhibitory effect on the endogenous proteases. Adhesive infiltration of resin-dentin interface and tensile bond strength before and after thermomechanical cycling were implemented to indicate that the PAMAM-OH pretreatment had no adverse effects on immediate dentin bonding and prolonged the durability of resin-dentin bonds. CONCLUSIONS: PAMAM-OH possesses anti-proteolytic activity and prevents exposed collagen fibrils within HLs from degradation, which lays the foundation for the satisfactory intrafibrillar remineralization induced by PAMAM-OH within HLs to achieve durable resin-dentin bonds in the next work.

Effect of Cavity Cleanser With Long-Term Antibacterial and Anti-Proteolytic Activities on Resin-Dentin Bond Stability.

Objective: Secondary caries caused by oral microbiome dysbiosis and hybrid layer degradation are two important contributors to the poor resin-dentin bond durability. Cavity cleansers with long-term antimicrobial and anti-proteolytic activities are in demand for eliminating bacteria-induced secondary caries and preventing hybrid layers from degradation. The objectives of the present study were to examine the long-term antimicrobial effect and anti-proteolytic potential of poly(amidoamine) dendrimers with amino terminal groups (PAMAM-NH2) cavity cleanser. Methods: Adsorption tests by attenuated total reflectance-infrared (ATR-IR) spectroscopy and confocal laser scanning microscopy (CLSM) were first performed to evaluate whether the PAMAM-NH2 cavity cleanser had binding capacity to dentin surface to fulfill its relatively long-term antimicrobial and anti-proteolytic effects. For antibacterial testing, Streptococcus mutans, Actinomyces naeslundii, and Enterococcus faecalis were grown on dentin surfaces, prior to the application of cavity cleanser. Colony-forming unit (CFU) counts and live/dead bacterial staining were performed to assess antibacterial effects. Gelatinolytic activity within the hybrid layers was directly detected by in situ zymography. Adhesive permeability of bonded interface and microtensile bond strength were employed to assess whether the PAMAM-NH2 cavity cleanser adversely affected resin-dentin bonding. Finally, the cytotoxicity of PAMAM-NH2 was evaluated by the Cell Counting Kit-8 (CCK-8) assay. Results: Adsorption tests demonstrated that the binding capacity of PAMAM-NH2 on dentin surface was much stronger than that of 2% chlorhexidine (CHX) because its binding was strong enough to resist phosphate-buffered saline (PBS) washing. Antibacterial testing indicated that PAMAM-NH2 significantly inhibited bacteria grown on the dentin discs as compared with the control group (p < 0.05), which was comparable with the antibacterial activity of 2% CHX (p > 0.05). Hybrid layers conditioned with PAMAM-NH2 showed significant decrease in gelatin activity as compared with the control group. Furthermore, PAMAM-NH2 pretreatment did not adversely affect resin-dentin bonding because it did not decrease adhesive permeability and microtensile strength. CCK-8 assay showed that PAMAM-NH2 had low cytotoxicity on human dental pulp cells (HDPCs) and L929. Conclusions: PAMAM-NH2 cavity cleanser developed in this study could provide simultaneous long-term antimicrobial and anti-proteolytic activities for eliminating secondary caries that result from a dysbiosis in the oral microbiome and for preventing hybrid layers from degradation due to its good binding capacity to dentin collagen matrix, which are crucial for the maintenance of resin-dentin bond durability.

Quaternary ammonium silane-based antibacterial and anti-proteolytic cavity cleanser.

OBJECTIVE: Secondary caries and degradation of hybrid layers are two major challenges in achieving durable resin-dentin bonds. The objectives of the present study were to investigate the effects of a 2% quaternary ammonium silane (QAS) cavity cleanser on bacteria impregnated into dentin blocks and the gelatinolytic activity of the hybrid layers. METHODS: Microtensile bond strength was first performed to evaluate if the 2% QAS cavity cleanser adversely affected bond strength. For antibacterial testing, Streptococcus mutans and Actinomyces naeslundii were impregnated into dentin blocks, respectively, prior to the application of the cavity cleanser. Live/dead bacterial staining and colony-forming unit (CFU) counts were performed to evaluate their antibacterial effects. Gelatinolytic activity within the hybrid layers was directly examined using in-situ zymography. A double-fluorescence technique was used to examine interfacial permeability immediately after bonding. RESULTS: The cavity cleanser did not adversely affect the bond strength of the adhesives tested (p>0.05). Antibacterial testing indicated that 2% QAS significantly killed impregnated bacteria within the dentin blocks compared with control group (p<0.05), which was comparable with the antibacterial activity of 2% chlorhexidine (p>0.05). Hybrid layers pretreated with 2% QAS showed significant decrease in enzyme activity compared with control group. With the use of 2% QAS, relatively lower interfacial permeability was observed, compared with control group and 2% chlorhexidine (p<0.05). SIGNIFICANCE: The present study developed a 2% QAS cavity cleanser that possesses combined antimicrobial and anti-proteolytic activities to extend the longevity of resin-dentin bonds.

Optimizing resin-dentin bond stability using a bioactive adhesive with concomitant antibacterial properties and anti-proteolytic activities.

Secondary caries and hybrid layer degradation are two major challenges encountered in long-term resin-dentin bond stability. As a link between resin and dentin, adhesives that possess both antimicrobial and anti-proteolytic activities are in demand for eliminating bacteria-induced secondary caries and preventing hybrid layers from degradation. In the present study, a new quaternary ammonium methacryloxy silane (QAMS) prepared from sol-gel chemistry was incorporated into experimental adhesives to examine their antimicrobial effect and anti-proteolytic potential. This functional methacrylate resin monomer contains polymerizable methacryloxy functionalities as well as a positively-charged quaternary ammonium functionality with a long, lipophilic -C18H37 alkyl chain for puncturing the cell wall/membrane of surface-colonizing organisms. Antibacterial testing performed using agar diffusion test, live/dead bacterial staining and colony-forming unit counts all indicated that the QAMS-containing adhesives killed Streptococcus mutans and Actinomyces naeslundii in a dose-dependent manner via a predominant contact-killing mechanism. Gelatinolytic activity within the hybrid layers created by these adhesives was examined using in-situ zymography. Hybrid layers created with 0% QAMS-containing adhesive exhibited intense green fluorescence emitted by the hydrolyzed fluorescein-conjugated gelatin, with 4-fold increase in enzymatic activity compared with an experimental adhesive containing 5% QAMS. Taken together, incorporation of 5% QAMS in the experimental adhesive provides simultaneous antimicrobial and anti-proteolytic activities that are crucial for the maintenance of long-term resin-dentin bond integrity. STATEMENT OF SIGNIFICANCE: Durability of resin-dentin interfacial bond remains a clinically-significant challenge. Secondary caries caused by bacteria and the degradation of hybrid layers via endogenous dentin proteases are two important contributors to the poor resin-dentin bond durability. The present study developed a new 5% QAMS-containing adhesive that provides simultaneous antimicrobial and dentin protease inhibition functions to extend the longevity of resin-dentin bonds.

Antimicrobial efficacy of an apical negative pressure root canal irrigation system against intracanal microorganisms.

OBJECTIVES: The effect of irrigation time on the antimicrobial efficacy of an apical negative pressure irrigation system was examined in vitro, followed by validation of the antimicrobial effect in vivo using the identified optimal irrigation time. METHODS: For the in vitro experiment, 44 extracted premolars were decoronated, instrumented, autoclaved and inoculated with Enterococcus faecalis (ATCC 29212) for 21 days. Four teeth were used as positive control, without irrigation. Each of the remaining 40 teeth was irrigated with 2.5% NaOCl, delivered via the EndoVac MacroCannula for 10 s, and subsequently via the EndoVac MicroCannula for 15, 30, 45, 60 or 90 s per canal, respectively (N = 8). After irrigation, microbial samples were collected, transferred to BHI broth and incubated for counting of bacterial colony forming units (CFUs). Based on the in vitro results, 8.25% NaOCl was delivered via the EndoVac MicroCannula for 60 s, during root canal treatment of 20 human subjects presented with apical periodontitis. Microbial samples retrieved in vivo prior to canal instrumentation (S0), after chemomechanical debridement (S1) and after irrigation with EndoVac (S2) were cultured in an anaerobic chamber for 7 days for CFU evaluation. RESULTS: Compared with the control, irrigation significantly reduced bacterial populations (p < .05). Irrigation delivery via the EndoVac demonstrated improved antibacterial efficacy with increased irrigation time (p < .05). Samples retrieved from canals after NaOCl delivery in vivo with the EndoVac for 60 s were all culture-negative. CONCLUSIONS: Microbial elimination may be achieved with 8.25% NaOCl delivered via the EndoVac apical negative pressure irrigation device for 60 s. CLINICAL SIGNIFICANCE: With the use of the EndoVac apical negative pressure irrigant delivery system, optimal elimination of the intracanal bacterial load can only be achieved when sodium hypochlorite is delivered via the MicroCannula for at least 60 s per canal.

Bioinspired heptapeptides as functionalized mineralization inducers with enhanced hydroxyapatite affinity.

The regeneration of mineral crystals under physiological conditions is an efficient way to repair defects in hard tissues. To achieve robust mineralization on surfaces such as the tooth enamel, an inducer requires strong affinity with the substrates and should be able to induce mineralization. Thus far, most studies used a single molecule containing two components to realize the above functions separately, which might be troublesome to synthesize and purify. In this work, inspired by the statherin in the salivary acquired pellicle, we designed a simple peptide sequence, Asp-Asp-Asp-Glu-Glu-Lys-Cys (peptide-7), to accomplish the dual tasks of adsorption and mineralization on enamel surfaces. We speculate the calcium binding ability of the negatively charged carboxylic acid groups in the peptide itself contributes to the dual functions of peptide-7. In vitro and in vivo experiments demonstrated its excellent repair effect on enamel as compared to fluoride. More importantly, due to the strong affinity between peptides and hydroxyapatite, a compact mineralized crystal layer and a strong adhesion between the regenerated minerals and the bottom substrates were observed, similar to the effect induced by fluoride. This work sheds light on the interaction mechanism between peptide-7 and minerals. In addition, since it is safer than fluoride, peptide-7 may have potential applications in the repair of other hard tissues and the functionalization of biomaterials.

Susceptibility of contemporary single-bottle self-etch dentine adhesives to intrinsic water permeation.

OBJECTIVES: To evaluate the effect of intrinsic water permeation on the microtensile bond strengths of different adhesive systems to dentine and the quality of resin-dentine interfaces. METHODS: Ninety-six non-carious human third molars were divided into 4 groups: Clearfil S3 Bond Plus (CSBP; Kuraray); Clearfil S3 Bond (C3S; Kuraray); iBond Self-Etch (IB; Heraeus-Kulzer) and Prime&Bond NT (PB, control etch-and-rinse adhesive, Dentply-Sirona). For each adhesive, specimens from one subgroup (N=10) were bonded using zero pulpal pressure, while specimens from the other subgroup (N=10) were bonded using 15cm water pressure (PP). Each bonded tooth was sectioned into 1×1mm sticks and stressed to failure. Data were analysed using two-way ANOVA and Holm-Sidak pairwise comparisons to examine the effects of "adhesive", "pulpal pressure" and their interaction on bond strength (α=0.05). Representative fractured sticks were examined by SEM. The remaining tooth slabs in each subgroup were used for TEM and CLSM. RESULTS: Microtensile bond strengths (mean±SD; in MPa) were: 33.4±6.9 (CSBP), 33.2±4.7 (CSBP-PP), 35.0±8.6 (C3S), 25.5±7.3 (C3S-PP), 18.4±4.0 (IB), 16.5±6.9 (IB-PP), 28.2±5.5 (PB), 20.5±7.2 (PB-PP). "Adhesive-type" (P<0.001), "pulpal-pressure" (P<0.001) and their interactions (P<0.001) significantly affected bond strength results. No difference between no-PP and PP subgroups was found for CSBP and IB (P>0.05). Water droplets were identified along the resin-dentine interface for IB, IB-PP and C3S-PP. CONCLUSION: IB exhibits water sensitivity when bonding is performed with/without pulpal pressure. C3S exhibits water sensitivity when bonding is performed with pulpal pressure. CSBP does not exhibit water sensitivity when bonding is performed with/without pulpal pressure. CLINICAL SIGNIFICANCE: Intrinsic water permeation during bonding procedures significantly affects bond strength results and the resin-dentine interface of contemporary single-bottle self-etch dentine adhesive systems.

Preliminary analysis of salivary microbiome and their potential roles in oral lichen planus.

Several studies have explored the origin and development mechanism of oral lichen planus (OLP) with limited attention to the role of bacteria in the progression of this common oral disease. Here we utilized MiSeq sequencing of 16S rRNA gene amplicons to identify complex oral microbiota associated with OLP from saliva samples of two subtypes (reticular and erosive) of OLP patients and healthy controls. Our analyses indicated that the overall structure of the salivary microbiome was not significantly affected by disease status. However, we did observe evident variations in abundance for several taxonomic groups in OLP. Porphyromonas and Solobacterium showed significantly higher relative abundances, whereas Haemophilus, Corynebacterium, Cellulosimicrobium and Campylobacter showed lower abundances in OLP patients, as compared with healthy controls. In addition, we explored specific microbial co-occurrence patterns in OLP, and revealed significantly fewer linkers of Streptococcus comprising species in erosive OLP. Furthermore, the disease severity and immune dysregulation were also genus-associated, including with Porphyromonas that correlated to disease scores and salivary levels of interleukin (IL)-17 and IL-23. Overall, this study provides a general description of oral microbiome in OLP, and it will be useful for further investigation of their potential roles in the initiation and immune modulation of OLP.

8DSS-promoted remineralization of demineralized dentin in vitro.

Dentin phosphoprotein (DPP) plays an extremely important role in the biomineralization of human tooth. The repetitive nucleotide sequence of aspartate-serine-serine (DSS) is the fundamental unit within DPP, and peptides containing 8 repeats of DSS (8DSS) have been shown to possess the ability to induce remineralization of the demineralized enamel. In this work, we coated the 8DSS peptide on the completely demineralized dentin to evaluate the effect of 8DSS peptide coating on dentin remineralization. Human dentin samples were demineralized with 37% phosphoric acid for 2 min, and then the 8DSS peptide (1 mg mL-1) was coated and the binding strength of the 8DSS peptide to demineralized dentin was examined using attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR). Then the coated dentin samples were immersed in artificial saliva for 3 weeks. After that, the remineralized dentin samples were characterized by scanning electron microscopy (SEM), energy dispersive spectroscopy (EDS), X-ray diffraction (XRD) analysis and ATR-FTIR. The mechanical properties of the remineralized surfaces were determined by nano-indentation and atomic force microscopy (AFM). The results showed that the 8DSS peptide had good binding strength to demineralized dentin and could induce nano-crystals precipitation both on the surfaces and within the dentinal tubules. The mechanical properties of the 8DSS-coated samples were significantly improved. In contrast, there were hardly any newly generated minerals deposited on the samples without 8DSS peptide coating. In conclusion, the 8DSS peptide may be a promising biomaterial for restoring the demineralized human dentin.

TCAB1: a potential target for diagnosis and therapy of head and neck carcinomas.

BACKGROUND: WRAP53, including α, ß and γ isoforms, plays an important role not only in the stability of p53 mRNA, but also in the assembly and trafficking of the telomerase holoenzyme. It has been considered an oncogene and is thought to promote the survival of cancer cells. The aim of this study was to detect the role of TCAB1 (except WRAP53α) in the occurrence and development of head and neck carcinomas. METHODS: Immunohistochemistry was used to detect the TCAB1 expression in clinical specimen sections and performed western blotting to check the TCAB1 expression levels in cell lines. TCAB1 was depleted using shRNA lentivirus and the knockdown efficiency was assessed using q-PCR and Western blotting. We performed CCK-8 assays and flow cytometry to check the cell proliferation potential and used the trans-well assay to test the invasion ability in vitro. Xenografts were used to detect the tumor formation potential in vivo. Moreover, we performed cDNA microarray to investigate the candidate factors involved in this process. RESULTS: We observed a notable overexpression of TCAB1 in head and neck carcinoma clinical specimens as well as in carcinoma cell lines. Knockdown of TCAB1 decreased the cellular proliferation potential and invasion ability in vitro. cDNA microarray analysis suggested the possible involvement of several pathways and factors associated with tumorigenesis and carcinoma development in the TCAB1-mediated regulation of cancers. Furthermore, the xenograft assay confirmed that the depletion of TCAB1 would inhibit tumor formation in nude mice. The immunohistochemistry results of the mice tumor tissue sections revealed that the cells in shTCAB1 xenografts showed decreased proliferation potential and increased apoptotic trend, meanwhile, the angiogenesis was inhibited in the smaller tumors form shTCAB1 cells. CONCLUSIONS: Our study demonstrated that depletion of TCAB1 decreased cellular proliferation and invasion potential both in vitro and in vivo. The data indicated that TCAB1 might facilitate the occurrence and development of head and neck carcinomas. In future, TCAB1 might be useful as a prognostic biomarker or a potential target for the diagnosis and therapy of head and neck carcinomas.

[Correlation between DNA damage and EB virus infection in nasopharyngeal carcinoma].

OBJECTIVE: To explore the correlation between the expression of phosphorylated histone H2AX (γ-H2AX) and Epstein-Barr virus (EBV) infection in nasopharyngeal carcinoma (NPC) specimens, and to investigate the role and mechanism of EBV-induced DNA damage in NPC tumorigenesis and development in vitro. METHODS: We enrolled 50 cases of NPC and 20 cases of nasopharyngitis (NPI) specimens to test the expressions of γ-H2AX and EBV latent membrane protein 1 (LMP1) by immunohistochemical method (IHC). And then in LMP1-negative samples, we detected the EBV-encoded RNA (EBER) using in situ hybridization. The γ-H2AX level was detected in NPC cells CNE1 before and after EBV infection using Western blotting. RESULTS: γ-H2AX was expressed in most NPC specimens (94%), which was much higher than that in NPI (40%), and EBV was presented in 94% of NPC but only 30% in NPI. Moreover, γ-H2AX was positive in 97.9% of the EBV-positive specimens, which indicated the close correlation between γ-H2AX expression and EBV infection (P<0.05). Finally, Western blotting showed that γ-H2AX level significantly increased in CNE1 cells after EBV infection. CONCLUSION: This study demonstrated that an intimate connection existed between γ-H2AX expression and EBV infection in NPC both in vivo and in vitro. EBV infection might induce DNA damage in CNE1 cells, which causes genome instability and initiates or promotes the tumorigenesis and development of NPC.

DNA binding properties and biological evaluation of dihydropyrimidinones derivatives as potential antitumor agents.

The binding properties of two medicinally important dihydropyrimidinones derivatives 5-(Ethoxycarbonyl)-6-methyl-4-phenyl-3,4-dihydropyrimidin-2(1H)-one (EMPD) and 5-(Ethoxycarbonyl)-6-methyl-4-(4-chlorophenyl)-3,4-dihydropyrimidin-2(1H)-one (EMCD) with calf-thymus DNA (ctDNA) were investigated by spectroscopy, viscosity, isothermal titration calorimetry (ITC) and molecular modeling techniques. Simultaneously, their biological activities were evaluated with MTT assay method. The binding constants determined with spectroscopic titration and ITC were found to be in the same order of 10(4)M(-1). According to the results of viscosity studies, fluorescence competitive binding experiment and ITC investigations, intercalative binding was evaluated as the dominant binding modes between the two compounds and ctDNA. Furthermore, the results of molecular modeling corroborated those obtained from spectroscopic, viscosimetric and ITC investigations. Evaluation of the antitumor activities of the two derivatives against different tumor cell lines proved that they exhibited significant tumor cell inhibition rate, accordingly blocking DNA transcription and replication. The present results favor the development of potential drugs related with dihydropyrimidinones derivatives in the treatment of some diseases.

A "turn-on" fluorescent sensor for detection of Pb2+ based on graphene oxide and G-quadruplex DNA.

A rapid, sensitive and selective fluorescence sensor for detection of Pb(2+) was developed based on a Pb(2+)-induced G-quadruplex and graphene oxide. By using a specific G-rich DNA sequence, this strategy provided a promising alternative to Pb(2+) determination in the presence of Hg(2+) escaping from the use of a masking agent of Hg(2+).