[Safety evaluation of Tibetan medicine Qishiwei Zhenzhu Pills based on serum pharmacochemistry and network pharmacology].

To explore the mechanism of the active ingredients of Qishiwei Zhenzhu Pills in inhibiting the hepatorenal toxicity of the zogta component based on serum pharmacochemistry and network pharmacology, thereby providing references for the clinical safety application of Qishiwei Zhenzhu Pills. The small molecular compounds in the serum containing Qishiwei Zhenzhu Pills of mice were identified by high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS). Then, by comprehensively using Traditional Chinese Medicines Systems Pharmacology(TCMSP), High-throughput Experiment-and Reference-guided Database(HERB), PubChem, GeneCards, SuperPred, and other databases, the active compounds in the serum containing Qishiwei Zhenzhu Pills were retrieved and their action targets were predicted. The predicted targets were compared with the targets of liver and kidney injury related to mercury toxicity retrieved from the database, and the action targets of Qishiwei Zhenzhu Pills to inhibit the potential mercury toxicity of zogta were screened out. Cytoscape was used to construct the active ingredient in Qishiwei Zhenzhu Pills-containing serum-action target network, and STRING database was used to construct the protein-protein interaction(PPI) network of intersection targets. The Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment analyses were carried out on the target genes by the DAVID database. The active ingredient-target-pathway network was constructed, and the key ingredients and targets were screened out for molecular docking verification. The results showed that 44 active compounds were identified from the serum containing Qishiwei Zhenzhu Pills, including 13 possible prototype drug ingredients, and 70 potential targets for mercury toxicity in liver and kidney were identified. Through PPI network topology analysis, 12 key target genes(HSP90AA1, MAPK3, STAT3, EGFR, MAPK1, APP, MMP9, NOS3, PRKCA, TLR4, PTGS2, and PARP1) and 6 subnetworks were obtained. Through GO and KEGG analysis of 4 subnetworks containing key target genes, the interaction network diagram of active ingredient-action target-key pathway was constructed and verified by molecular docking. It was found that taurodeoxycholic acid, N-acetyl-L-leucine, D-pantothenic acid hemicalcium, and other active ingredients may regulate biological functions and pathways related to metabolism, immunity, inflammation, and oxidative stress by acting on major targets such as MAPK1, STAT3, and TLR4, so as to inhibit the potential mercury toxicity of zogta in Qishiwei Zhenzhu Pills. In conclusion, the active ingredients of Qishiwei Zhenzhu Pills may have a certain detoxification effect, thus inhibiting the potential mercury toxicity of zogta and playing a role of reducing toxicity and enhancing effect.

KCNN4-mediated Ca2+/MET/AKT axis is promising for targeted therapy of pancreatic ductal adenocarcinoma.

As a member of the potassium calcium-activated channel subfamily, increasing evidence suggests that KCNN4 was associated with malignancies. However, the roles and regulatory mechanisms of KCNN4 in PDAC have been little explored. In this work, we demonstrated that the level of KCNN4 in PDAC was abnormally elevated, and the overexpression of KCNN4 was induced by transcription factor AP-1. KCNN4 was closely correlated with unfavorable clinicopathologic characteristics and poor survival. Functionally, we found that overexpression of KCNN4 promoted PDAC cell proliferation, migration and invasion. Conversely, the knockdown of KCNN4 attenuated the growth and motility of PDAC cells. In addition to these, knockdown of KCNN4 promoted PDAC cell apoptosis and led to cell cycle arrest in the S phase. In mechanistic investigations, RNA-sequence revealed that the MET-mediated AKT axis was essential for KCNN4, encouraging PDAC cell proliferation and migration. Collectively, these findings reveal a function of KCNN4 in PDAC and suggest it's an attractive therapeutic target and tumor marker. Our studies underscore a better understanding of the biological mechanism of KCNN4 in PDAC and suggest novel strategies for cancer therapy.

A systematic review: differentiation of stem cells into functional pericytes.

Pericytes are an integral cellular component of vascular structures. Numerous studies have investigated various stem cell types as potential sources of pericytes for application in cell-based therapy. The diverse stem cell types and variable experimental protocols of these studies make it imperative to evaluate the relevant scientific literature on the basis of a unified standard. The purpose of this systematic review is to rigorously evaluate the relevant scientific literature for conclusive evidence that stem cells can differentiate into functional pericytes. An online literature search was conducted up to July 2016. Eligible papers were evaluated on 4 pertinent criteria: 1) appropriate controls, 2) markers to confirm pericyte phenotype, 3) techniques for assessing pericyte functionality, and 4) differentiation efficiency of the protocol. Our search yielded 20 eligible studies (from 2006 to 2016), 12 of which were published in the past 5 yr. Of these 20 articles, only 1 had positive control, and 5 papers evaluated differentiation efficiency. The most commonly used pericyte markers were neuron-glial antigen 2, platelet-derived growth factor receptor-ß, and α-smooth muscle actin. Three articles were associated with adipose stem cells, 4 with mesenchymal stem cells, and 7 with pluripotent stem cells, whereas the remaining 6 articles were based on other miscellaneous stem cell types. Stem cells can serve as a potential source of pericytes, but there should be standardized guidelines in future studies for assessing pericyte differentiation.-Xu, J., Gong, T., Heng, B. C., Zhang, C. F. A systematic review: differentiation of stem cells into functional pericytes.

[Disinfection effects of MTAD on apical isthmus of the root canal system: a scanning electron microscopic evaluation].

OBJECTIVE: To evaluate the disinfection efficacy of MTAD on Enterococcus faecalis biofilm and smear layer colonization in apical isthums of the root canal system. METHODS: Fifteen extracted human maxillary first premolars with isthmus anatomic structure which confirmed by stereo-microscope were contaminated with E. faecalis in vitro and randomly divided into 5 groups: the first group was not treated serving as a baseline control, the second group was treated by normal saline (NS) serving as negative control, the third group was treated by MTAD , the forth group by 5.25% NaOCl, and the fifth group by 5.25% NaOCl + EDTA. All roots in the latter four groups were instrumented by Protaper rotary files and irrigated with respective irrigant, then the roots were split longitudinally and a scanning electron microscope was used to evaluate the antibacterial activity and smear layer cleaning ability of irrigants on isthmus. RESULTS: In the first group, E. faecalis colonized on the isthmus surface and aggregated together to form biofilm-like microorganism community, some bacteria also colonized in the dentinal tubules. When treated with NS, both smear layer and bacteria remained (median of smear layer score was 5). MTAD can remove partial smear layer, and have limited antibacterial activity, some bacteria embedded in smear layer (the median was 3) and were destroyed; In 5.25% NaOCl treatmentgroup, the smear layer was not removed (median of smear layer score was also 5), but all bacteria on the surface were extinguished. The combined use of 5.25% NaOCl and EDTA produced a cleaner isthmus surface and had marked antimicrobial effect, with the median of smear layer score being only 1. CONCLUSION: MTAD may permeate into the isthmus area of apical root canal system, but only performed a partial effect of disinfection and limited antibacterial activity. Sodium hypochlorite cooperated with EDTA can remove infection effectively in the isthmus area.

Meta-analysis Flavonoids from traditional Chinese herbs for diabetes in rats: a network Meta-analysi.

OBJECTIVE: In this Meta-analysis, we evaluated the hypoglycemic effect of 5 flavonoids found in traditional Chinese herbs (naringenin, kaempferol, puerarin, baicalein, and luteolin) on diabetic rats. METHODS: Four databases including PubMed, Web of Science, Embase, and Cochrane Library, were searched from inception to May 2020. Only studies using diabetes model rats were included in the analysis. Blood glucose data from the last measurement were collected and analyzed. Pair-wise Meta-analyses were conducted using STATA v14.0 software and a Meta-analysis was conducted using STATA v14.0, ADDIS v1.16.6, and R v3.6.1. The quality of included studies was assessed with the SYRCLE risk of bias tool for animal studies, and publication bias was evaluated with a comparisonadjusted funnel plot. RESULTS: A total of 33 studies were included in the analysis, in which all 5 flavonoids showed a beneficial effect on blood glucose level of diabetic rats were included in the final analysis. The standardized mean differences (95% confidence intervals) were -4.92 (-6.67, -3.17) fornaringenin, -12 (-18.74, -5.27) for kaempferol, -2.52 (-3.77, -1.26) for puerarin, -3.04 (-5.75, -0.34) for baicalein, and -1.94 (-2.95, -0.92) for luteolin. The network Meta-analysis showed no statistically significant differences between the effect sizes of the flavonoids. CONCLUSION: The results of the Meta-analysis showed that naringenin, kaempferol, puerarin, baicalein, and luteolin all have clear hypoglycemic effects in rat diabetes models, highlighting their therapeutic potential for preventing and treating diabetes mellitus in clinical practice.

Stem Cells from Human Exfoliated Deciduous Teeth (SHEDs) and Dental Pulp Stem Cells (DPSCs) Display a Similar Profile with Pericytes.

BACKGROUND: Pericytes play an important role in forming functional blood vessels and establishing stable and effective microcirculation, which is crucial for vascular tissue engineering. The slow ex vivo expansion rate, limited proliferative capacity, and variability of tissue-specific phenotypes would hinder experimental studies and clinical translation of primary pericytes. In this study, the angiogenic and pericyte functions of stem cells from human exfoliated deciduous teeth (SHEDs) and postnatal human dental pulp stem cells (DPSCs) were investigated. METHODS: Osteogenic and adipogenic induction assays were performed to evaluate the mesenchymal potential of SHEDs, DPSCs, and pericytes. An in vitro Matrigel angiogenesis assay was conducted to reveal the ability of SHEDs, DPSCs, and pericytes to stabilize vascular-like structures. Quantitative real-time polymerase chain reaction (RT-qPCR) was performed to evaluate mRNA expression. Flow cytometry, western blotting, and immunostaining were used to assess the protein expression. Wound healing and transwell assays were performed to evaluate the migration ability of SHEDs, DPSCs, and pericytes. RESULTS: The osteogenic and adipogenic induction assays showed that SHEDs, DPSCs, and pericytes exhibited similar stem cell characteristics. The mRNA expression levels of PDGFR-ß, α-SMA, NG2, and DEMSIN in SHEDs and DPSCs cultured in EC medium were significantly higher than those in the control groups on day 7 (P < 0.05), but significantly higher than those in the pericytes group on day 14 (P < 0.05). Flow cytometry showed that high proportions of SHEDs and DPSCs were positive for various pericyte markers on day 7. The DPSCs, SHEDs, and pericytes displayed strong migration ability; however, there was no significant difference among the groups (P > 0.05). CONCLUSION: The SHEDs and DPSCs display a profile similar to that of pericytes. Our study lays a solid theoretical foundation for the clinical use of dental pulp stem cells as a potential candidate to replace pericytes.

Mechanisms for Improving Hepatic Glucolipid Metabolism by Cinnamic Acid and Cinnamic Aldehyde: An Insight Provided by Multi-Omics.

Cinnamic acid (AC) and cinnamic aldehyde (AL) are two chemicals enriched in cinnamon and have been previously proved to improve glucolipid metabolism, thus ameliorating metabolic disorders. In this study, we employed transcriptomes and proteomes on AC and AL treated db/db mice in order to explore the underlying mechanisms for their effects. Db/db mice were divided into three groups: the control group, AC group and AL group. Gender- and age-matched wt/wt mice were used as a normal group. After 4 weeks of treatments, mice were sacrificed, and liver tissues were used for further analyses. Functional enrichment of differentially expressed genes (DEGs) and differentially expressed proteins (DEPs) were performed using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases. DEPs were further verified by parallel reaction monitoring (PRM). The results suggested that AC and AL share similar mechanisms, and they may improve glucolipid metabolism by improving mitochondrial functions, decreasing serotonin contents and upregulating autophagy mediated lipid clearance. This study provides an insight into the molecular mechanisms of AC and AL on hepatic transcriptomes and proteomes in disrupted metabolic situations and lays a foundation for future experiments.

Cyclic Adenosine Monophosphate Promotes Odonto/Osteogenic Differentiation of Stem Cells from the Apical Papilla via Suppression of Transforming Growth Factor Beta 1 Signaling.

INTRODUCTION: Stem cells from the apical papilla (SCAPs) possess strong odonto/osteogenic differentiation potential. This study investigated the effect of cyclic adenosine monophosphate (cAMP) on odonto/osteogenic differentiation of SCAPs and the underlining interplay between cAMP and transforming growth factor beta 1 (TGF-ß1). METHODS: SCAPs were stimulated with an activator of cAMP (forskolin) in the presence of either TGF-ß1 or a TGF-ß1 inhibitor. The amounts of calcium mineral deposition and alkaline phosphatase activity were determined. Quantitative real-time polymerase chain reaction was performed to elucidate cAMP on the TGF-ß1-mediated odonto/osteogenic differentiation of SCAPs. The effect of cAMP on the phosphorylation of Smad2/Smad3 and extracellular-regulated kinase (ERK)/P38 induced by TGF-ß1 was analyzed by Western blotting. RESULTS: Cotreatment with forskolin and a TGF-ß1 inhibitor enhanced alkaline phosphatase activity and deposition of calcium minerals in SCAPs. Moreover, the TGF-ß1 inhibitor synergized the effect of forskolin on the expression of type I collagen and runt-related transcription factor 2. The results of Western blotting revealed that forskolin attenuated the unregulated expression of the phosphorylation of Smad3 and ERK induced by TGF-ß1, and a cAMP inhibitor (H89) antagonized this effect. CONCLUSIONS: This study showed that cAMP signaling exerts its up-regulating effects on the odonto/osteogenic differentiation of SCAPs by interfering with TGF-ß1 signaling via inhibiting Smad3 and ERK phosphorylation.

[Expression of receptor activator of nuclear factor kappaB ligand and osteoprotegerin in chronic apical periodontitis].

OBJECTIVE: To investigate the expression of key mediators that regulate bone resorption, receptor activator of nuclear factor-kappaB ligand (RANKL) and osteoprotegerin (OPG) in human periapical granuloma (PG) and radicular cyst (RC) characterized by periapical bone destruction. METHODS: Immunohistochemical technique was performed using monoclonal antibody to detect RANKL and OPG expression in PG (n= 20 ) and RC (n= 20 ). As control group, healthy periodontal ligament tissues (n = 6 ) were obtained from teeth with orthodontic indication of extraction. RESULTS: The RANKL protein expression was significantly higher in PG (43.74 +/- 8.40) and RC (40.33 +/- 7.53) than in control group (15.47 +/- 2.59, P=0.000) , but no statistical significance could be found between PG and RC groups(P=0.161) .The expression of OPG in PG (27.81 +/- 5.17), RC (26.35 +/- 3.86) and control group (24.33 +/- 3.50) show no statistical significance (P >0.05). Moreover, RANKL/OPG ratio in PG (1.59 +/- 0.26) and RC (1.54 +/- 0.24) was much higher than that in control group (0.64 +/- 0.10, P=0.000), however the ratio of RANKL/OPG showed no significant difference between PG and RC groups (P= 0.504) . CONCLUSION: RANKL and OPG were observed in the periapical lesions. RANKL may be responsible for bone resorption in periapical lesions. RANKL and OPG may play an important role in the development of periapical lesions.

Evaluation of the bactericidal effect of Er,Cr:YSGG, and Nd:YAG lasers in experimentally infected root canals.

The aim of this study was to evaluate the bactericidal effect of the Er,Cr:YSGG laser and the Nd:YAG laser in experimentally infected root canals. Sixty single-rooted teeth with straight canals were selected. After preparation and sterilization, the specimens were inoculated with Enterococcus faecalis for 3 weeks. After irradiation by lasers, the number of bacteria in each root canal was examined. The Er,Cr:YSGG laser gave a reduction of 77% after irradiation at 1 W and 96% at 1.5 W, but there was no significant difference (p > 0.05). The Nd:YAG laser gave a reduction of 97% at 1 W and 98% at 1.5 W, and there was no significant difference (p > 0.05). Compared with the Er,Cr:YSGG laser, the Nd:YAG laser is more effective (p < 0.05). In conclusion, both lasers systems have a significant bactericidal effect in infected root canals, and the Nd:YAG laser is more effective than the Er,Cr:YSGG laser.

TGF-ß1-induced differentiation of SHED into functional smooth muscle cells.

BACKGROUND: Adequate vascularization is crucial for supplying nutrition and discharging metabolic waste in freshly transplanted tissue-engineered constructs. Obtaining the appropriate building blocks for vascular tissue engineering (i.e. endothelial and mural cells) is a challenging task for tissue neovascularization. Hence, we investigated whether stem cells from human exfoliated deciduous teeth (SHED) could be induced to differentiate into functional vascular smooth muscle cells (vSMCs). METHODS: We utilized two cytokines of the TGF-ß family, transforming growth factor beta 1 (TGF-ß1) and bone morphogenetic protein 4 (BMP4), to induce SHED differentiation into SMCs. Quantitative real-time polymerase chain reaction (RT-qPCR) was used to assess mRNA expression, and protein expression was analyzed using flow cytometry, western blot and immunostaining. Additionally, to examine whether these SHED-derived SMCs possess the same function as primary SMCs, in vitro Matrigel angiogenesis assay, fibrin gel bead assay, and functional contraction study were used here. RESULTS: By analyzing the expression of specific markers of SMCs (α-SMA, SM22α, Calponin, and SM-MHC), we confirmed that TGF-ß1, and not BMP4, could induce SHED differentiation into SMCs. The differentiation efficiency was relatively high (α-SMA+ 86.1%, SM22α+ 93.9%, Calponin+ 56.8%, and SM-MHC+ 88.2%) as assessed by flow cytometry. In vitro Matrigel angiogenesis assay showed that the vascular structures generated by SHED-derived SMCs and human umbilical vein endothelial cells (HUVECs) were comparable to primary SMCs and HUVECs in terms of vessel stability. Fibrin gel bead assay showed that SHED-derived SMCs had a stronger capacity for promoting vessel formation compared with primary SMCs. Further analyses of protein expression in fibrin gel showed that cultures containing SHED-derived SMCs exhibited higher expression levels of Fibronectin than the primary SMCs group. Additionally, it was also confirmed that SHED-derived SMCs exhibited functional contractility. When SB-431542, a specific inhibitor of ALK5 was administered, TGF-ß1 stimulation could not induce SHED into SMCs, indicating that the differentiation of SHED into SMCs is somehow related to the TGF-ß1-ALK5 signaling pathway. CONCLUSIONS: SHED could be successfully induced into functional SMCs for vascular tissue engineering, and this course could be regulated through the ALK5 signaling pathway. Hence, SHED appear to be a promising candidate cell type for vascular tissue engineering.

Partial Pulpotomy of Immature Teeth with Apical Periodontitis using Bioceramics and Mineral Trioxide Aggregate: A Report of Three Cases.

Pulpal necrosis of an immature permanent tooth with an open apex poses a challenge for the clinician. The conventional apexification technique using calcium hydroxide has yielded short-term success, but this technique has inevitable shortcomings. Hence, this case series aimed to evaluate the effectiveness of using bioceramics (iRoot BP) or mineral trioxide aggregate (MTA) for partial pulpotomies. Three boys aged 9 to 11 years old presented with partial pulp necrosis and symptomatic apical periodontitis of the mandibular right and left second premolar. The involved teeth were treated with a partial pulpotomy using either iRoot BP (case 1 and 2) or MTA (case 3). At the 8-month follow-up, no abnormal clinical signs or symptoms were observed. Periapical radiographs revealed a significant reduction in periapical radiolucency, a marked increase in the root canal wall thickness and ongoing closure of the apical opening. The bioceramic material (iRoot BP) and MTA both produced successful outcomes in the partial pulpotomy of immature teeth with partial pulp necrosis and apical periodontitis. However, iRoot BP was superior in terms of ease of clinical application, and would therefore be a better treatment alternative than MTA.

Perceptions of single-visit and multiple-visit endodontic treatment: a survey of endodontic specialists and general dentists in Hong Kong.

AIM: To study the preference of practice for single- and multiple-visit endodontic treatment by Hong Kong endodontists and general dental practitioners (GDPs), and to investigate their reasons for choosing single- or multiple-visit treatment in their practice. METHOD: An anonymous questionnaire was mailed to all 16 registered endodontists and 800 randomly selected GDPs in Hong Kong to explore their preference and reasons for selecting single- or multiple-visit endodontic treatment for their patients. Information on the use of magnifying loupes, microscopes and the number of years they have been in dental practice was also collected. RESULTS: Eight endodontists and 429 GDPs returned their questionnaires and the response rate was 50% and 53.6% respectively. Among the GDPs, 404 (94.2%) undertook endodontic treatment in their practices. For those performing endodontic treatment, the mean number of years of practice was 23.6 ± 4.8 for endodontists and 15.3 ± 9.1 for GDPs. Seven endodontists (87.5%) used a surgical microscope. For GDPs, only 25 (6.2%) used a surgical microscope and 123 (30.4%) used magnifying loupes during endodontic treatment. Seven endodontists (87.5%) and 375 GDPs (92.8%) predominantly performed multiple-visit treatment. The commonest reasons for choosing multiple-visit treatment for both endodontists and GDPs were the positive effects of interappointment medications (n = 3, 37.5%) and that the tooth to be treated had doubtful prognosis (n = 103, 25.5%). The commonest reason for choosing single-visit treatment for both endodontists and general dentists was that treatment could be completed in one visit (n = 4, 50%) and (n = 127, 31.4%). CONCLUSION: Most Hong Kong endodontists and GDPs preferred offering multiple-visit endodontic treatment.

The spherical nanoparticle-encapsulated chlorhexidine enhances anti-biofilm efficiency through an effective releasing mode and close microbial interactions.

We reported two forms (sphere and wire) of newly fabricated chlorhexidine (CHX)-loaded mesoporous silica nanoparticles (MSNs), and investigated their releasing capacities and anti-biofilm efficiencies. The interactions of the blank MSNs with planktonic oral microorganisms were assessed by field emission scanning electron microscopy. The anti-biofilm effects of the two forms of nanoparticle-encapsulated CHX were examined by 2,3-bis (2-methoxy- 4-nitro-5-sulfo-phenyl)-2H-tetrazolium-5-carboxanilide. The profiles of biofilm penetration were analyzed by fluorescent-labeled MSNs using confocal microscopy and ImageJ. The spherical MSNs with an average diameter of 265 nm exhibited a larger surface area and faster CHX-releasing rate than the MSN wires. The field emission scanning electron microscopy images showed that both shaped MSNs enabled to attach and further fuse with the surfaces of testing microbes. Meanwhile, the nanoparticle-encapsulated CHX could enhance the anti-biofilm efficiency with reference to its free form. Notably, the spherical nanoparticle-encapsulated CHX presented with a greater anti-biofilm capacity than the wire nanoparticle-encapsulated CHX, partly due to their difference in physical property. Furthermore, the relatively even distribution and homogeneous dispersion of spherical MSNs observed in confocal images may account for the enhanced penetration of spherical nanoparticle-encapsulated CHX into the microbial biofilms and resultant anti-biofilm effects. These findings reveal that the spherical nanoparticle-encapsulated CHX could preferably enhance its anti-biofilm efficiency through an effective releasing mode and close interactions with microbes.

Cellular Interactions and Formation of an Epithelial "Nanocoating-Like Barrier" with Mesoporous Silica Nanoparticles.

Oral mucosa as the front-line barrier in the mouth is constantly exposed to a complex microenvironment with multitudinous microbes. In this study, the interactions of mesoporous silica nanoparticles (MSNs) with primary human gingival epithelial cells were analyzed for up to 72 h, and their diffusion capacity in the reconstructed human gingival epithelia (RHGE) and porcine ear skin models was further assessed at 24 h. It was found that the synthesized fluorescent mesoporous silica nanoparticles (RITC-NPs) with low cytotoxicity could be uptaken, degraded, and/or excreted by the human gingival epithelial cells. Moreover, the RITC-NPs penetrated into the stratum corneum of RHGE in a time-dependent manner, while they were unable to get across the barrier of stratum corneum in the porcine ear skins. Consequently, the penetration and accumulation of RITC-NPs at the corneum layers of epithelia could form a "nanocoating-like barrier". This preliminary proof-of-concept study suggests the feasibility of developing nanoparticle-based antimicrobial and anti-inflammatory agents through topical application for oral healthcare.

[Coculture of stem cells from the apical papilla and human umbilical vein endothelial cells enhances the angiogenic potential of dental pulp tissues].

PURPOSE: To observe the effect of coculture of stem cells from the apical papilla (SCAPs) and human umbilical vein endothelial cells (HUVECs) on the angiogenic potential of dental pulp tissues. METHODS: SCAPs were incubated in osteo/odontogenic, adipogenic, neurogenic induction medium and α-MEM medium, whose multilineage differentiation capacities were confirmed using alizarin red staining, oil red O staining and ßIII-tubulin immunofluorescent staining. The tubular length, branching points number and junctional areas were detected after 3, 6, 9 h since cells were seeded onto matrigel, and the data was analyzed using SPSS 16.0 software package. RESULTS: SCAPs in the experimental groups were detected having more lipid droplets, mineralization nodules and neuron-like cells. Coculture of SCAPs and HUVECs formed more vessel-like structures in tubular formation assay. CONCLUSIONS: SCAPs are capable of differentiating into fat, bone, and nerve-like cells in vitro. Coculture of SCAPs and HUVECs can enhance the angiogenic potential of dental pulp tissues.

Morphological analysis of apical foramen over-instrumented by three rotary NiTi systems.

OBJECTIVE: To evaluate morphological changes of the apical surface after root canal preparation with 1 mm beyond the apical foramen using ProTaper Universal (PTU) files, K3 files and Twisted files (TF), respectively. METHODS: Seventy teeth with a centered apical foramen and 70 teeth with a deviated apical foramen were included as group A and group B respectively. In each group, 20 teeth were randomly assigned for root canal preparation with PTU, K3 and TF files, respectively; the remaining 10 teeth were used as the control group without any preparation. The apical foramens were examined with scanning electronic microscopy. The foramen integrity damage (FID) and dentin defects (DDs) were noted and compared between different groups. RESULTS: FID and DD were significantly less in Group A. DDs was not found in the control group. Preparation with PTU, K3, and TF files caused FID in 6.67%, 10%, and 3.33% of teeth in the group A, and in 20%, 26.67%, and 10% in Group B, respectively. Preparation with PTU, K3, and TF files caused DD in 6.67%, 6.67%, and 3.33% of teeth in Group A, and in 23.33%, 26.67%, and 6.67% in Group B, respectively. PTU and K3 files produced more DDS than TF files. However, no significant difference was found between groups using PTU and K3 files. CONCLUSION: Rotary instrumentation caused less damage on the apical surface in foramencentered root canals than foramen-deviated root canals when working beyond the canal length. TF files had a tendency to produce less DDS compared with PTU or K3 files during over-instrumented root canals.

Endodontic bacteria from primary and persistent endodontic lesions in Chinese patients as identified by cloning and 16S ribosomal DNA gene sequencing.

BACKGROUND: Few literatures pertain to the 16S ribosomal DNA (16S rDNA) analysis of bacteria contributing to primary and persistent endodontic lesions, with no information available for the Chinese population. As such, we investigated endodontic bacteria associated with primary and persistent endodontic lesions in adult Chinese patients living in Beijing, China using 16S rDNA gene sequencing techniques. METHODS: Endodontic microbial samples were obtained from fourteen adult Chinese patients and subjected to DNA extraction. Pllymerase chain reaction (PCR) products were cloned and 100 clones from each generated library were randomly selected. Purified plasmid DNA with 16S rDNA gene inserts was sequenced, and the sequences were searched against GenBank databases using the BLASTN algorithm. Only significant identification with the highest-scored BLAST result and 99% minimum similarity was considered for phylotyping. RESULTS: More than 150 taxa were obtained. Primary endodontic infection was mainly associated with Burkholderia cepacia, Actinomyces, Aranicola spp. and Streptococcus sanguinis, whilst Burkholderia cepacia was predominant in the persistent endodontic infections. CONCLUSION: There is a difference in the species profile associated with endodontic infections of Chinese patients living in Beijing in comparison to other geographical or ethnic reports.

Prevalence of Enterococcus faecalis in saliva and filled root canals of teeth associated with apical periodontitis.

To investigate the prevalence of Enterococcus faecalis in saliva and filled root canals of patients requiring endodontic retreatment for apical periodontitis. Patients with apical periodontitis who were referred for endodontic retreatment were examined. The type and quality of the restoration, symptoms, quality of obturation were recorded. During retreatment, an oral rinse sample and root canal sample were cultured using brain-heart infusion agar and bile esculinazide agar to select for E. faecalis. The 16S rRNA technique was used to identify E. faecalis. A total of 32 women and 22 men (mean age: 38 years; s.d.: 11 years) and 58 teeth were studied. The prevalence of E. faecalis was 19% in the saliva and 38% in the root canals. The odds that root canals harbored E. faecalis were increased if the saliva habored this bacterium (odds ratio=9.7; 95% confidence interval=1.8-51.6; P<0.05). Teeth with unsatisfactory root obturation had more cultivable bacterial species in root canals than teeth with satisfactory root obturation (P<0.05). E. faecalis is more common in root canals of teeth with apical periodontitis than in saliva. The prevalence of E. faecalis in root canals is associated with the presence of E. faecalis in saliva.

Malocclusions in Xia Dynasty in China.

BACKGROUND: The prevalence of malocclusion in modern population is higher than that in the excavated samples from the ancient times. Presently, the prevalence of juvenile malocclusion in the early stage of permanent teeth is as high as 72.92% in China. This study aimed to observe and evaluate the prevalence and severity of malocclusions in a sample of Xia Dynasty in China, and to compare these findings with the modern Chinese population. METHODS: The material consisted of 38 male and 18 female protohistoric skulls of Xia Dynasty 4000 years ago. Of 86 dental arches, 29 cases had the jaw relationships. Tooth crowding, diastema, individual tooth malposition and malocclusion were studied. RESULTS: Of the samples, 23.3% showed tooth alignment problems including crowding (8.1%), diastema (9.3%), and individual tooth malposition (5.8%). The prevalence of malocclusion was 27.6%, mainly presented as Angle Class I. CONCLUSIONS: It is indicated that over thousands of years from Neolithic Age (6000 - 7000 years ago) to Xia Dynasty (4000 years ago), the prevalence of malocclusion did not change significantly. The prevalence of malocclusion of Xia Dynasty samples was much lower than that of modern population.