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Substance P (SP) is a neuropeptide released from the nervous fibers in response to injury. In addition to its association with pain and reactions to anxiety and stress, SP exerts various physiological functions by binding to the neurokinin-1 receptor (NK1R). However, the expression and role of SP in reparative dentinogenesis remain elusive. Here, we explored whether SP is involved in odontoblastic differentiation during reparative dentinogenesis.Dental pulp stem cells (DPSCs) were isolated from healthy human dental pulp tissues and subjected to odontoblastic differentiation. The expression of SP and NK1R during odontoblastic differentiation was investigated in vitro. The effects of SP on odontoblastic differentiation of DPSCs were evaluated using alizarin red staining, alkaline phosphatase staining, and real-time polymerase chain reaction. After direct pulp capping with mineral trioxide aggregate (MTA), the expression of SP and NK1R during reparative dentin formation in rats were identified using histological and immunohistochemical staining.SP and NK1R expression increased during the odontoblastic differentiation of DPSCs. SP translocated to the nucleus when DPSCs were exposed to differentiation medium. NK1R was always present in the nuclei of DPSCs and odontoblast-like cells. Additionally, we discovered that 10SP contributes to odontoblastic differentiation during reparative dentin formation by binding to the NK1R.

Introduction Regenerative endodontics has created a desirable shift in the treatment paradigm despite current limitations of regenerative outcomes. Mesenchymal stem cells (MSCs) facilitate tissue regeneration and repair in a mild inflammatory environment. Small extracellular vesicles (sEVs) derived from MSCs play an imperative role in the paracrine modulation of regenerative responses modulated by MSCs. However, it remains unknown whether MSCs enhance dental pulp regeneration or whether this enhancement is mediated by sEVs in a mild inflammatory environment. The present study aimed to elucidate the effects of sEVs originated from lipopolysaccharide (LPS)-preconditioned human dental pulp stem cells (hDPSCs) on dental pulp regeneration. Methods All sEVs were isolated from hDPSCs cultured with or without LPS (ie, N-sEVs and L-sEVs, respectively). The effect of N-sEVs and L-sEVs on proliferation, migration, angiogenesis, and differentiation of rat bone marrow MSCs was identified in vitro. Moreover, N-sEVs or L-sEVs were implanted into rat pulpless root canal models, and the regenerated tissue in root canals was assessed via hematoxylin-eosin staining, Masson staining, and immunohistochemistry after 30 days of transplantation. Results Both N-sEVs and L-sEVs could modulate BMSC proliferation, migration, angiogenesis, and differentiation. Both kinds of sEVs enhanced the structure of the regenerated tissue closer to that of a normal dental pulp in vivo. L-sEVs had a more significant effect than N-sEVs. Conclusions sEVs released by hDPSCs in a mild inflammatory microenvironment are capable of facilitating the regeneration of dental pulp through functional healing instead of scar healing, which has potential applications in regenerative endodontics.

Macrophages have been reported to play important roles in tissue repair and regeneration. While it is known that macrophages are present in the dental pulp, their role in dental pulp regeneration is not fully understood. In the present study, we investigated the effects of different phenotype macrophages conditioned medium on the cellular behaviors of hDPSCs and their extracellular matrix (cell sheets) in vitro. Moreover, twenty-four root fragments inserted with cell sheets cultured with different conditioned media were placed into the back subcutaneous space of 6-8-week-old male BALB/c nude mouse. The regenerated tissues in the root fragments were assessed via histologic analysis after 8 weeks of transplantation. M2 macrophages could promote the proliferation, migration, and osteogenic differentiation of hDPSCs. Dental pulp-like tissue with an odontoblast-like layer lining the dentinal surface and well-arranged collagen fibers was harvested in root fragment combined with M2 conditioned medium cultured cell sheet, whereas a large amount of calcium salt deposition and disorganization of collagen fibers were observed in root fragments combined with M1 conditioned medium cultured cell sheet. Therefore, promoting the transformation of M1 into M2 macrophage in dental pulp tissue regeneration may be a potential way for dental pulp regeneration via functional healing.

Objective: The aim of this meta-analysis was to critically assess the effect of cochlear implantation on auditory and speech performance outcomes of children with auditory neuropathy spectrum disorder (ANSD). Material and methods: A systematic literature search was conducted on PubMed, EMbase, and Web of Science. The outcomes included speech recognition score, Categories of Auditory Performance (CAP), Speech Intelligibility Rating (SIR) score, and open-set speech perception. Results were expressed as standardized mean difference (SMD) or risk ratio (RR) with a 95% confidence interval (95% CI). Results: A total of 15 studies was included in this meta-analysis. Pooled data showed that, there were no significant differences between ANSD and sensorineural hearing loss (SNHL) groups in terms of speech recognition score (SMD = 0.01, 95% CI: −0.45, 0.47; P = .959),CAP (SMD = 0.71, 95% CI: −0.13, 1.54; P = .098), SIR score (SMD = −0.09, 95% CI: −0.49, 0.32; P = .667), and open-set speech perception (RR = 0.85, 95% CI: 0.69, 1.05; P = .142). Sensitivity analysis by removing individual studies one at a time showed that the overall estimate and level of heterogeneity did not change substantially. Conclusion: The current evidence suggested that children with ANSD who underwent cochlear implants achieved comparable effects in auditory and speech performance as children with non-ANSD SNHL.

Background and Aim. Nonalcoholic fatty liver disease (NAFLD) is an independent risk factor for cardiovascular disease. Hepatic fibrosis is the most significant determinant of all-cause- and liver -related mortality in NAFLD. However, the relationship between NAFLD fibrosis and severe coronary artery disease (CAD) remains unclear. Methods and Results. We conducted a retrospective study of 531 patients with ultrasonogram-confirmed NAFLD who underwent percutaneous coronary intervention (PCI). Then, all patients were separated into four categories by Gensini score (0, 0-9, 9-48, and ≥48) for use in ordinal logistic regression analysis to determine whether NAFLD fibrosis was associated with increased Gensini scores. Mediation analysis was used to investigate whether systemic inflammation is a mediating factor in the association between NAFLD fibrosis and CAD severity. FIB − 4 > 2.67 ( OR = 5.67 , 95% CI 2.59-12.38) and APRI > 1.5 ( OR = 14.8 , 95% CI 3.24-67.60) remained to be independent risk factors for the severity of CAD after adjusting for conventional risk factors, whereas among the inflammation markers, only neutrophils and neutrophil-to-lymphocyte ratio (NLR) were independently associated with CAD. Multivariable ordinal regression analysis suggested that increasing Gensini score (0, 0-9, 9-48, and ≥48) was associated with advanced NAFLD fibrosis. ROC curve showed that either fibrosis markers or inflammation markers, integrating with traditional risk factors, could increase the predictive capacity for determining CAD. Inflammation markers, especially neutrophils and NLR, were mediators of the relationship between NAFLD fibrosis and CAD severity. Conclusions. NAFLD patients with advanced fibrosis are at a high risk of severe coronary artery stenosis, and inflammation might mediate the association between NAFLD fibrosis and CAD severity.

The detection of precise hearing thresholds in infants and children with auditory neuropathy (AN) is challenging with current objective methods, especially in those younger than six months of age.The aim of this study was to compare the thresholds using auditory steady-state response (ASSR) and cochlear microphonics (CM) in children with AN and children with normal hearing.The thresholds of CM, ASSR, and visual reinforcement audiometry (VRA) tests were recorded; the ASSR and VRA frequencies used were 250, 500, 1000, 2000, and 4000 Hz.The participants in this study were 15 children with AN (27 ears) (1–7.6 years, median age 4.1 years) and ten children with normal hearing (20 ears) (1–8 years, median age four years).The thresholds of the three methods were compared, and histograms were used to represent frequency distributions of threshold differences obtained from the three methods.In children with normal hearing, the average CM thresholds (84.5 dB) were significantly higher than the VRA thresholds (10.0–10.8 dB); in children with AN, both CM and VRA responses were seen at high signal levels (88.9 dB and 70.6–103.4 dB, respectively). In normal children, the difference between mean VRA and ASSR thresholds ranged from 17.5 to 30.3 dB, which was significantly smaller than the difference seen between the mean CM and VRA thresholds (71.5–72.3 dB). The correlation between VRA and ASSR in children with normal hearing ranged from 0.38 to 0.48, whereas no such correlation was seen in children with AN at any frequency (0.03–0.19).Our results indicated that ASSR and CM were poor predictors of the conventional behavioral threshold in children with AN.

BackgroundReports have proven that shorter door-to-needle time (DTN time) indicates better outcomes in AIS patients received intravenous thrombolysis. Efforts have been made by hospitals and centers to minimize DTN time in many ways including introducing a stroke nurse. However, there are few studies to discuss the specific effect of stroke nurse on patients’ prognosis. This study aimed to compare consecutive AIS patients before and after the intervention to analyze the effect of stroke nurse on clinical outcome of AIS patients.MethodsIn this retrospective study, we observed 1003 patients from November 2016 to December 2020 dividing in two groups, collected and analyzed AIS patients’ medical history, clinical assessment information, important timelines, 90 mRS score, etc. Comparative analysis and mediation analysis were also used in this study.ResultsA total of 418 patients was included in this study, and 199 patients were enrolled in the stroke nurse group and 219 was in the preintervention group. Baseline characteristics of patients showed no significant difference except there seems more patients with previous ischemic stroke history in the group of stroke nurse. (p = 0.008). The median DTN time significantly decreased in the stroke nurse group (25 min versus 36 min,p p = 0.001). Mediation analysis indicated the reduction of DTN time plays a partial role on the 90 days mRS score and the stroke nurse has some direct effect on the improvement of clinical outcome (p = 0.006).ConclusionsThe introduction of stroke nurse is beneficial to clinical outcome of AIS patients and can be use of reference in other hospitals or centers.

Qi-Wen Deng,1,&ast; Shi Huang,1,&ast; Shuo Li,2,&ast; Qian Zhai,1 Qing Zhang,1 Zhen-Jie Wang,1 Wen-Xia Chen,1 Huiling Sun,3 Min Lu,1 Junshan Zhou1 1Department of Neurology, Nanjing First Hospital, Nanjing Medical University, Nanjing, 210006, People’s Republic of China; 2Department of Neurology, Affiliated ZhongDa Hospital, School of Medicine, Southeast University, Nanjing, 210009, People’s Republic of China; 3General Clinical Research Center, Nanjing First Hospital, Nanjing Medical University, Nanjing, 210006, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Junshan Zhou; Min Lu Department of Neurology, Nanjing First Hospital, Nanjing Medical University, 210006, People’s Republic of China Email zhjsh333@126.com; luminnfhsjk@126.comBackground and Purpose: This study aimed to explore several peripheral blood-based markers related to the inflammatory response in a total of 210 patients with acute ischemic stroke (AIS) caused by large artery occlusion in the anterior circulation who received endovascular therapy (EVT) from an observational study of clinical significance of circulating non-coding RNA in acute ischemic stroke (AISRNA).Methods: We collected baseline characteristics of 210 AIS patients participating in an observational acute stroke cohort: the AISRNA study. The following inflammatory factors were measured in these participants: interleukin-2 [IL-2], IL-4, IL-6, IL-10, tumor necrosis factor-α [TNF-α], and interferon-γ [IFN-γ]. The National Institute of Health Stroke Scale score increase of ≥ 4 within 24 hours after EVT defined as early neurological deterioration (END).Results: Compared with patients without END, patients with END had a higher incidence of atrial fibrillation (P=0.012), and also had higher levels of IL-6 and IL-10 (P< 0.01). Furthermore, we found that the area under the curves (AUCs) of IL-6 and IL-10 for predicting END were 0.768 (0.697– 0.829), and 0.647 (0.570– 0.719), respectively. Adjusting for age, sex, and atrial fibrillation, the odds ratios (ORs; 95% confidence interval) for incident END for IL-6 and IL-10 were 1.98 (1.05– 6.69) and 1.18 (1.04– 1.33), respectively. Additionally, we found significant changes over time in the expression levels of IL-4, IL-6, and IL-10 in patients with END compared with patients without END (P< 0.05).Conclusion: IL-6 and IL-10 levels at admission may be potential markers of END after EVT, and the time course of IL-4, IL-6, and IL-10 is correlated with stroke progression. Further larger studies are needed to confirm the current findings.Trial Registration: ClinicalTrials.gov NCT04175691. Registered November 21, 2019, https://www.clinicaltrials.gov/ct2/show/NCT04175691Keywords: acute ischemic stroke, early neurological deterioration, endovascular therapy, inflammatory factors, IL-6

BackgroundBiallelic mutations in LOXHD1 have been identified as the cause of DFNB77 (deafness, autosomal recessive 77). It is a novel, progressive, severe-profound, and late-onset non-syndromic hearing loss, and is genetically and phenotypically highly heterogeneous. This study aimed to provide an additional three cases of DFNB77 to analyze this complex disease.MethodsWe presented three cases of pediatric patients with prelingual milder form of the DFNB77 with residual hearing at low frequencies. Trio whole-exome sequencing (WES) was conducted to identify the pathogenic variants. Additionally, we reviewed the literature to further analyze the relationships between the genotype and audiology phenotype of LOXHD1 worldwide.ResultsSix novel possible pathogenic LOXHD1 variants in three patients were identified by WES, including three missense, one nonsense, and two splicing variants. The literature review showed that 68.5% of DFNB77 patient onset before five years old; Most variants (60%) were associated with a milder phenotype, particularly variants in the protein domain of PLAT 7 and PLAT 9. We found that compared with homozygous LOXHD1 variants, individuals with heterozygous compound variants had a significantly milder phenotype, especially individuals carrying one missense and one splicing or bi-allelic missense variants (P <0.05). Audiometric analysis at different ages showed that the hearing loss degree was aggravated at all frequencies in adulthood and more severe in elderhood.Conclusions We report three children with hearing loss carrying six novel LOXHD1 variants identified by WES. Furthermore, our work indicates that DFNB77 may be milder than previously reported, and recommends considering the genotype combination and mutation location of LOXHD1 and race-specificity in DFNB77 molecular diagnoses and management.

Background: This study aimed to explore several peripheral blood-based markers related to inflammatory response in a total of 85 patients with acute ischemic stroke (AIS) caused by large artery occlusion in the anterior circulation receiving endovascular therapy (EVT) from the AISRNA study regarding the association between inflammatory factors and early neurological deterioration (END), and investigated whether their time course correlated with END after EVT.Methods: We collected baseline characteristics of 85 AIS patients participating in an observational acute stroke cohort: the AISRNA study. The following inflammatory factors were measured in these participants: interleukin-2 [IL-2], IL-4, IL-6, IL-10, tumor necrosis factor-α [TNF-α], and interferon-γ [IFN-γ]. The National Institute of Health Stroke Scale score increase of ≥4 within 24 hours after EVT defined as END.Results: IL-6 and IL-10 were higher in patients with END compared to those with non-END (PPConclusions: IL-6 and IL-10 levels on admission are significantly associated with END after EVT, and time course of IL-4, IL-6, and IL-10 is correlated with stroke progression. Further study of molecular mechanisms on peripheral immunomodulation in AIS would be helpful.Trial registration: ClinicalTrials.gov NCT04175691. Registered November 21, 2019.

Autosomal recessive deafness-102 (DFNB102), a new profound prelingual non-syndromic hearing loss, is caused by mutations in the EPS8 gene. To date, only three such consanguineous families with three different homozygous variants in EPS8 have been reported. Here, we report the fourth case from a non-consanguineous Chinese family, an 11-month-old male infant presented with congenital profound non-syndromic hearing loss. Trio whole-exome sequencing initially identified the patient with a novel seemingly homozygous splicing variant NM_004447.5: c.1435-2A > T in intron 14 of the EPS8 gene and was inherited from his father; further CNVs analysis identified a novel 65.9 kb intragenic deletion and was inherited from his mother. The deletion is covering intron 14 that could account for the apparent homozygosity of the patient. In vitro splicing assay showed the variant c.1435-2A > T creates a new donor site at position c.1443, which is predicted to produce a stop codon after 14 additional amino acids (p.His479Cysfs*14). Furthermore, quantitative allele-specific expression assay showed that relative EPS8 gene expression in the patient significantly decreased (0-fold for the wild-type transcript and 0.25-0.27-fold for the mutant transcript) compared to the control (P < 0.05), indicating the pathogenicity of the identified variants. Overall, our study provides additional evidence that EPS8 is a causative gene for DFNB102 and highlights the clinical utility of simultaneous analysis of CNVs and SNVs to avoid potential errors in the diagnosis and interpretation of patients with apparent homozygosity.

Autoantibodies against tumor-associated antigens (TAAs) have been found in many kinds of cancers, and might serve as biomarkers for early cancer diagnosis. The present study was carried out to test if there is any relation between autoantibodies against Fascin and esophageal squamous cell carcinoma (ESCC).One hundred and forty-nine patients with ESCC and 98 control subjects were recruited in the study. The levels of circulating autoantibodies against Fascin were measured by enzyme-linked immunosorbent assay (ELISA). Receiver operating characteristic (ROC) was used to calculate diagnostic accuracy.The levels of autoantibodies against Fascin in patients with ESCC were significantly higher than in control subjects (P0.001). Measurement of autoantibodies against Fascin provided an area under the curve (AUC) of 0.636, [95% confidence interval (CI): 0.568-0.704] 24.8% sensitivity (95% CI: 18.3%-37.2%) and 99.0% specificity (95% CI: 93.6%-99.9%). Moreover, serum level of autoantibodies against Fascin in early-stage ESCC was significantly higher than that of normal controls (P0.05). The positive rates of autoantibodies against Fascin were correlated with age (P0.05), but not with gender, tumor size, tumor site, histological grade, T stage, N stage or TNM stage (P0.05).Our results suggest that Fascin autoantibody may be a potential biomarker for the early detection of ESCC.

Epitaxial ${\mathrm{BiMnO}}_{3}$ (BMO) thin films were prepared on (001)-oriented ${\mathrm{SrTiO}}_{3}$ substrates with a ${\mathrm{SrRuO}}_{3}$ buffer layer using pulsed laser deposition. The correlation between ferroelectric and structural properties of BMO has been investigated in detail. By high-resolution transmission electron microscopy measurement, we observed a structural transition from pseudocubic to monoclinic structure with increasing thickness of BMO films, which can be ascribed to the intermediate layer and small composition fluctuation, i.e., Bi deficiency. The ferroelectricity of the BMO films has been characterized by piezoresponse force microscope, demonstrating that pseudocubic BMO thin films show intrinsic ferroelectricity, while the ferroelectricity is absent or very weak in the monoclinic BMO thin films at room temperature. The results of density-functional theory calculations also support the experimental results.

Biallelic mutations in LOXHD1 have been identified as the cause of DFNB77 (deafness, autosomal recessive 77). It is a new progressive, severe-to-profound, and late-onset nonsyndromic sensorineural hearing loss (NSHL), and is highly heterogeneous genetically and phenotypically. This study aimed to provide an additional three cases of DFNB77.We presented three unrelated children diagnosed with prelingual mild-to-severe NSHL, and their audiograms showed mild hearing loss at 250 Hz before downsloping to a moderate-to-severe degree. Trio whole-exome sequencing (WES) was conducted to identify the pathogenic variants. Additionally, we reviewed the literature to further analyze the relationships between the genotype and audiology phenotype of LOXHD1.Six novel possible pathogenic LOXHD1 variants were identified, including three missense, one nonsense, and two splicing variants. The literature review showed that 68.5% of patients with DFNB77 onset before five years old; Most variants (62%) were associated with a down-sloping audiogram of mild-to-moderate hearing loss at low frequencies (200Hz, 500Hz), particularly variants in the protein domain of PLAT 9. We found that compared with homozygous LOXHD1 variants, individuals with heterozygous compound variants had a significantly milder phenotype, especially individuals carrying one missense and one splicing or bi-allelic missense variants (P 0.05). Audiometric analysis at different ages showed that the hearing loss degree was aggravated at all frequencies by increasing age.We report three children with prelingual NSHL carrying six novel LOXHD1 variants. Furthermore, our work indicates that DFNB77 may be milder than previously reported and recommends considering the genotype combination and mutation location of LOXHD1 and race-specificity in DFNB77 molecular diagnoses and management.

Introduction Platelet-rich fibrin (PRF) has been used as a scaffold material in various tissue regeneration studies. In the previous methods to combine seed cells with PRF, the structure of PRF was damaged, and the manipulation time in vitro was also increased. The objective of this in vitro study was to explore an appropriate method to develop a PRF–human dental pulp cell (hDPC) complex to maintain PRF structure integrity and to find out the most efficient part of PRF. Methods The PRF-hDPC complex was developed at 3 different time points during PRF preparation: (1) the before centrifugation (BC) group, the hDPC suspension was added to the venous blood before blood centrifugation; (2) the immediately after centrifugation (IAC) group, the hDPC suspension was added immediately after blood centrifugation; (3) the after centrifugation (AC) group, the hDPC suspension was added 10 minutes after blood centrifugation; and (4) the control group, PRF without hDPC suspension. The prepared PRF-hDPC complexes were cultured for 7 days. The samples were fixed for histologic, immunohistochemistry, and scanning electron microscopic evaluation. Real-time polymerase chain reaction was performed to evaluate messenger RNA expression of alkaline phosphatase and dentin sialophosphoprotein. Enzyme-linked immunosorbent assay quantification for growth factors was performed within the different parts of the PRF. Results Histologic, immunohistochemistry, and scanning electron microscopic results revealed that hDPCs were only found in the BC group and exhibited favorable proliferation. Real-time polymerase chain reaction revealed that alkaline phosphatase and dentin sialophosphoprotein expression increased in the cultured PRF-hDPC complex. The lower part of the PRF released the maximum quantity of growth factors. Conclusions Our new method to develop a PRF-hDPCs complex maintained PRF structure integrity. The hDPCs were distributed in the buffy coat, which might be the most efficient part of PRF.

The gap junction β2 (GJB2) gene is associated with more than half of the recessive forms of hereditary hearing loss (HHL). However, the correlation between p.V27I and p.E114G variants of GJB2 and hearing phenotype remains controversial. This study aimed to clarify possible roles of these variants in Chinese infants with nonsyndromic hearing loss (NSHL). Hearing and gene tests were conducted in 300 infants (aged 0-3 months) with NSHL and 484 normal infants (aged 0-3 months). The p.V27I and p.E114G variants appeared frequently in both NSHL patients and normal controls. The allele and haplotype frequencies of p.V27I and p.E114G in patients and controls were compared, but no significant difference was observed (p=0.44 and p=0.26, respectively). Moreover, genotype frequencies of the p.V27I variant showed no significant difference between the two groups (p=0.66). Interestingly, more homozygote p.V27I/p.E114G subjects were found in NSHL infants than in controls (5/484 and 13/300, respectively), most of whom (61.54%) had mild or moderate hearing losses. Our results indicate that homozygote p.V27I/p.E114G is associated with mild and moderate HHL.

The aim of this study was to compare the quality of root fillings completed by a modified single-cone (MSC) technique with 3 different sealers after minimal instrumentation and multisonic cleaning of root canals of maxillary first molars.Root canals of 18 maxillary first molars were instrumented to size 15/.04 taper using rotary files. Sodium hypochlorite 5.25% was used during instrumentation; the final cleaning was performed by the GentleWave System (Sonendo Inc, Laguna Hills, CA). The specimens were allocated into 3 groups and root filled by the MSC technique using a size fitted gutta-percha master cone and GuttaFlow Bioseal (Coltene Whaledent GmBH + Co KG, Langenau, Switzerland), GuttaFlow 2 (Coltene Whaledent GmBH + Co KG), and MTA Fillapex (Angelus Industria de Produtos Odontológicos S/A, Londrina, PR, Brazil) sealers. Micro-computed tomographic scans were obtained before and after instrumentation, post-GentleWave, and after obturation. Reconstructed images were analyzed for the volumetric percentage of filling materials. Mesiobuccal roots of the selected teeth were sectioned at 0.5-mm increments starting at the apex of the root. The cross sections were further examined using a light microscope.The 3 groups had 90%-99% of the canal space filled with the root filling material. The mean volume of the filling material was higher in the GuttaFlow Bioseal and GuttaFlow 2 groups than in the MTA Fillapex group (P.05). There was no significant difference among the apical, middle, and coronal thirds. The cross-sectional images showed no obvious gaps or voids in the GuttaFlow groups. After instrumentation, 49 of the 189 canal thirds (25.9%) had hard tissue debris in the root canal system. After GentleWave cleaning, only 4 of 63 canals (6.3%) and 4 of the 189 canal thirds (2.1%) still had debris.The MSC method with GuttaFlow 2 and GuttaFlow Bioseal sealers after multisonic cleaning of minimally instrumented molar canals resulted in high-quality root fillings. Multisonic cleaning of minimally instrumented molars seems to be effective in debris removal.

ObjectiveTo study the effect of lipoteichoic acid (LTA) and 5-FU on the expression of caspase-3, EGFR, TGF-α proteins of tumor tissue of H22 cancer bearing mice and its anti-tumor mechanism.MethodsA total of 40 SPF grade Kunming mice were selected to establish H22 liver cancer model, and then the mice were divided into 4 groups at random with ten mice in each group. Group A was given saline lavage treatment, Group B was treated with 5-FU by intraperitoneal injection, Group C was treated with LTA by lump body injection; Group D was treated with LTA by lump body injection and 5-FU by intraperitoneal injection. Two weeks after the treatment, the mice in each group were executed and the tumor tissue was stripping and weighted, and the tumor growth inhibition ratio was calculated. Then the tumor tissue was processed for conventional embedding, sectioned to observe the expression of caspase-3, EGFR, TGF-α by immunohistochemical staining method.ResultsThe tumor inhibitory rate o f Group D was significantly higher than Groups B and C (P 0.05). The IDO values of TGF-α, EGFR proteins in Groups B, C, D mice tumor tissue were significantly lower than that in group A (P

Introduction Dentinogenesis includes odontoblast differentiation and extracellular matrix maturation as well as dentin mineralization. It is regulated by numerous molecules. High-temperature requirement protein A1 (HtrA1) plays crucial roles in bone mineralization and development and is closely associated with the transforming growth factor beta (TGF-β) signal in osteogenesis differentiation. Simultaneously, the TGF-β1/small mother against decapentaplegic (Smad) signaling pathway is an important signaling pathway in various physiological processes and as a downstream regulation factor of HtrA1. However, the role of HtrA1 and its relationship with the TGF-β1/Smad signaling pathway in dentin mineralization is unknown. Methods We detected the role of HtrA1 and its relationship with the TGF-β1/Smad signaling pathway in odontoblastic differentiation of human dental pulp cells (hDPCs) in this study. First, hDPCs were cultured in mineralized medium, and odontoblastic differentiation was confirmed by investigating mineralized nodule formation, alkaline phosphatase (ALP) activity, and the expression of mineral-associated genes, including ALP, collagen I, and dentin sialophosphoprotein. Then, the expression of HtrA1 and TGF-β1/Smad in hDPCs was investigated in hDPCs during mineralized induction. After HtrA1 knockdown by lentivirus, the mineralized nodule formation, ALP activity, and expression of mineral-associated genes and TGF-β1/Smad genes were investigated to confirm the effect of HtrA1 on odontoblastic differentiation and its relationship with the TGF-β1/Smad signaling pathway. Results The expression of HtrA1 and TGF-β1 was increased during odontoblastic differentiation of hDPCs along with the messenger RNA expression of downstream factors of the TGF-β1/Smad signaling pathway. In addition, lentivirus-mediated HtrA1 knockdown inhibited the process of mineralization and the expression of HtrA1 and TGF-β1/Smad genes. Conclusions These findings suggest that HtrA1 might positively regulate odontoblastic differentiation of hDPCs through activation of the TGF-β1/Smad signaling pathway.

Esophageal squamous cell carcinoma (ESCC), one of the most common malignancies worldwide, is a highly aggressive and homogeneous entity occurring in esophageal squamous epithelium, and a reliable noninvasive test for early detection is needed. A recent study showed that serum autoantibodies against Ezrin could be detected in patients with pancreatic cancer. Here, we assessed whether autoantibodies against Ezrin could have diagnostic relevance for early ESCC. We analyzed autoantibodies against Ezrin in sera of 98 normal controls and 149 patients with ESCC. Ezrin autoantibodies levels were evaluated by enzyme-linked immunosorbent assay (ELISA). Results showed that higher levels of autoantibodies against Ezrin were observed in serum samples from patients with ESCC than in serum from normal controls (P<0.0001). Based on a cutoff value of 0.319, the sensitivity and specificity of autoantibodies against Ezrin for diagnosis of ESCC were 27.5% and 95.9%, respectively. Compared with normal controls, the positive rate of autoantibodies against Ezrin was significantly elevated in patients with early-stage ESCC (P<0.0001). Moreover, there was no significant difference of positivity of autoantibodies against Ezrin in ESCC patients categorized according to age, gender, tumor size, tumor invasion depth, tumor site, histological grade, lymph node status, or tumor stage. Our study indicates that the presence of autoantibodies against Ezrin is significantly associated with ESCC.

This study was designed to analyze the three-dimensional structure of platelet-rich fibrin (PRF) gel as scaffold for dental pulp regeneration, and to investigate the effect of PRF gel on the proliferation of human dental pulp cells (hDPCs) in vitro.PRF gel prepared by Choukroun's protocols was evaluated by light microscope and scanning electron microscope (SEM). The extract of PRF gel was collected at the time point of 7th day. The experimental group was divided into 2 subgroups (25% and 75%) according to the volume fraction of PRF gel extract. The 2 subgroups were named 1PRF and 3PRF, respectively. The hDPCs were isolated and cultured in DMEM with 1PRF and 3PRF, respectively. Cell proliferation was detected using cell counting kit-8 (CCK-8) at 24, 48, 72, 96, 120, 144 and 168 h, respectively. The data was analyzed with SPSS16.0 software package.Platelet and leukocyte were mainly distributed at the junction between the red corpuscles and the PRF gel, which was called "buffy coat". "Buffy coat" was constructed by large and dense fibrin clusters. At 24, 48, 72 and 96 h, there was no significant difference (OD) value of optical density(OD) between experimental group and control group(P0.05). At 120, 144 and 168 h, the OD value in the experimental group was significant higher than that of the control group (P0.05); there was no significant difference of OD value between group 1PRF and 3PRF (P0.05).The results confirm that the three-dimensional structure of PRF gel is constituted by fibrin network. A large amount of platelet and leukocyte which can release high quantities of growth factors are concentrated in the PRF gel. The extract of PRF gel can accelerate the proliferation of hDPCs in time-dependent manner. PRF gel may be a proper scaffold for pulp regeneration. Supported by National Natural Science Foundation of China (81160133).

To compare the cleanliness of root end preparations by using ultrasonic instrumentation and slow-speed handpiece.Thirty-two mesial roots of the first mandibular molars with two canals and mature root apices were assigned randomly to 2 groups, each group had 16 teeth. The root-end preparations were made respectively using ultrasonic diamond tip Berutti and NiTi tip RE2 and slow-speed handpiece with No.2 round bur. Root end cavities were examined under scanning electron microscope for further evaluation of the superficial debris and smear layer of the root end preparations. SPSS 13.0 software package was used for Kruskal Wallis test.Ultrasonic preparation had significantly less superficial debris and smear layer than slow-speed handpiece preparation (P0.05).Ultrasonic instrument creates cleaner surfaces for root end cavities than slow-speed handpiece preparation in posterior teeth root end preparation.

To compare the apical sealing ability of the two filling materials: Resilon/Epiphany Self-Etch and Gutta-percha/AH plus.Thirty-six recently extracted mandibular premolar roots were included, 6 for the positive and negative groups, 28 for the experimental groups (14 for the Resilon/Epiphany SE group, 14 for the Gutta-percha / AH plus group).The roots were prepared according to the same standards, then the root canals were filled respectively with Resilon/Epiphany SE and Gutta-percha/AH plus. The teeth were put into the India ink to stain, and the apical sealing ability of the two filling materials: Resilon/Epiphany Self-Etch and Gutta-percha/AH plus were compared with the methods of dye leakage. SPSS13.0 software package was used for data analysis.The sealer of the Resilon/Epiphany SE was bonding to the resin filling material and the root canal dentin, and became a structure named mono-block. Compared with the Gutta-percha /AH plus group, the Resilon/Epiphany SE group had significantly lower apical leakage(P0.05).The results show that the Resilon/Epiphany SE filling system has better apical sealing ability than the traditional filling material of Gutta-percha/AH plus, and the sealer of Resilon/Epiphany SE is bonded to the resin filling materials and the root canal dentin, and become a structure named monoblock.

To compare the therapeutic efficacy both apexification and revascularization in the immature anterior teeth of animal model with apical periodontitis, and observe the histological situation of revascularization in the root canal.Six immature anterior teeth of one animal model (dog) aged approximately 4.5 months was selected. Afterwards, periapical periodontitis pattern were established, the samples were randomly divided into the experimental group (revascularization, 3 teeth) and the control group (apexification, 3 teeth). To compare the development of root and the healing of periapical inflammation, the involved teeth were respectively radiographed 1, 4, 8 weeks after surgery. The animals were sacrificed after 8 weeks, and the closure of apical foramen and the content of root canal were observed by hematine-eosine (HE) staining.The postoperative radiography after 1 week and 4 weeks, the apical foramen size and the periapical radiolucency of the samples was shown no perceptual change. After 8 weeks, the experimental group periapical radiolucency area was obviously more narrowing, and had a apical closure tendency whereas the thickness of the root canal walls had imperceptible changed. While the control group periapical radiolucency change varied. The granulation tissue could be seen within the lumen of the experimental group, which contained a large number of irregular calcification, the calcification was obvious in the apical and adjacent the root canal wall. A small quantity of hard tissue was deposited in the apical of the control group.Revascularization may increase the recovery of immature anterior teeth with chronic periapical inflammation, the vital regenerative tissue within root canal is the granulation tissue contained calcification.

To evaluate the effects of retrograde preparation between ultrasonic retrotips and traditional slow-speed handpiece.27 maxillary second bicuspid teeth with two canals and one isthmus were divided randomly into 3 groups with 9 in each. The root-end preparations were made respectively using ultrasonic diamond tip Berutti and NiTi tip RE2, ultrasonic #40K file and slow-speed handpiece with NO.2 round bur. The postoperative epoxy resin replicas were examined under scanning electron microscope for further evaluating the number and type of the micro-cracks, the quality of cavity margin. SPSS13.0 software package was used for Kruskal-Wallis test.The number of micro-crack of ultrasonic instruments were significantly less than that of slow-speed handpiece(P=0.002).The type of micro-cracks has no significant difference between the two techniques(P=0.657). On the quality of cavity margin, the slow-speed handpiece group had significant inferior to the ultrasonic #40K file group(P=0.039); there was no significantly difference between the ultrasonic Berutti and RE2 group and the slow-speed handpiece group (P=0.136); there was no significant difference between the two ultrasonic groups (P=0.637).Ultrasonic instrument is feasible in the root-end preparation.