Objective:To investigate the possibility of using long-wave carbon dots to trace rat bone marrow mesenchymal stem cells (BMSCs) and its effects on cell activity and osteogenic differentiation. Methods:CDs were characterized by TEM and FT-IR. CCK8 assay and flow cytometry were applied to screen the optimal concentration of CDs for imaging. After the CDs were co-cultured with BMSCs, the pathways of cells up taking CDs were investigated through a variety of transmembrane inhibitors and culturing at 4 ℃ low temperature. laser scanning confocal microscopy was used to trace the labeled cells for 9 days. Lysosome fluorescent probe was used to observe the distribution of CDs. ALP activity was measured at 4 days and 7 days, mineralization ability was detected by alizarin red staining at 21 days, and the osteogenic markers Runx2, Ocn, Opn, Bmp2 and Osterix were detected by PCR at 7 days and 14 days. Results:TEM showed that CDs were spherical particles and the average particle size was 2.17 mm. Ft-IR analysis showed that the surface of CDs was rich in hydroxyl and amino groups. CCK-8 results showed that the cell activity increased significantly when the concentration of CDs was 50 and 100 μg/mL at the 1st day. When the concentration reached 300 μg/mL, the cell activity decreased, but was still higher than 80%. Flow cytometry results showed that the labeling efficiency increased with the increase of CDs concentration. The red fluorescence of CDs overlapped partially with the green fluorescence of lysosomes and partially located in the cytoplasm, but did not enter the nucleus. The fluorescence could still be seen after 9 days. When the energy-dependent endocytosis pathway was inhibited at 4 ℃, the uptake amounts of CDs in the cells were significantly decreased. Compared with the control group, the ALP activity, mineralization ability and the expressions of Runx2, Ocn, Opn, Bmp2, Osterix were all increased in the CDs group. Conclusions:CDs have the potential to trace BMSCs and regulate the osteogenic differentiation.

Objective:?To explore the distribution characteristics of primary cilia in tooth development and further reveale the correlation between primary cilia and tooth development. Methods:?Hematoxylin-eosin (HE) staining and immunofluorescence staining of histological sections were used to observe the distribution characteristics of primary cilia in the bud, cap, and bell stages of mouse tooth germ development, as well as the matrix secretion period after birth. Results:?The primary cilia were widely distributed in the bud stage, while rarely observed in the basement membrane of the epithelium; cells with primary cilia in inner enamel epithelium gradually increased at the cap and bell stages; In one-week mouse tooth germs, all ameloblasts in the secretory phase had primary cilia, and the location of primary cilia in these cells was switched to the matrix secretion side, while there are fewer primary cilia in the odontoblasts during the same period. Conclusions:?Primary cilia have different distribution characteristics at different stages of tooth germ development; the appearance and location of primary cilia in ameloblasts are closely related to the differentiation of ameloblasts; primary cilia are dynamically involved in the whole process of mouse tooth development.

Objective:To investigate the effect of Rho GEF Trio on neural crest cells during embryonic development. Methods:Trio function was inhibited by microinjection of morpholino (MO) into zebrafish embryos，the development of mandibular and neural crest cells derivative was observed at 96 h through stereo microscope, and the development of craniomaxillofacial cartilage was observed at 120 h by Alcian blue staining. The effect of trio on the neural crest cells migration path of zebrafish was observed by using the neural crest cell specific labeling technique of transgenic zebrafish (sox10：EGFP)by means of Timelapse. Trio knockdown neural crest cells were cultured in vitro to observe the effect of Trio on the migration of neural crest cells through measuring the wound area (%) after migration in wound healing assay. Results:When trio function was inhibited，zebrafish mandibular development was insufficient, the Meckel's cartilage became retrusive, and the distance of neural crest cells migration decreased. In vitro，the wound area after neural crest cell migration increased in the Trio knockdown group. Conclusions:Trio may promote craniofacial development of neural crest origin by regulating the neural crest cells migration.

Objective:To prepare the polycaprolactone(PCL)/type I collagen(COLI)/nano-calcium zirconate(nCZ) composite scaffold for bone tissue regeneration, and to evaluate its performance, biocompatibility and growth of human periodontal ligament cells (PDLCs) effects of bone differentiation. Methods:The PCL/COLI, PCL/COLI/nHA and PCL/COLI/nCZ scaffolds were prepared by the electrospinning technique. The structure and properties of the scaffolds were observed by scanning electron microscope (SEM), the element composition of the stent was analyzed by an energy dispersive X-ray spectrometer (EDS), and the mechanical properties of the stent were tested by a universal stretching machine. Co-cultivation of the scaffolds was done with PDLCs, CCK-8 method was used to detect cell proliferation, cell morphology was observed by SEM, alkaline phosphatase (ALP) staining and Alizarin red (ARS) staining were used to observe the mineralization ability of each group of scaffolds on PDLCs. Results:The PCL/COLI/nCZ composite scaffold had a porous network structure, and the surface of the composite scaffold with nanomaterials had a rough appearance. The tensile test showed that the PCL/COLI/nCZ composite scaffold had the best mechanical strength among the three composite scaffolds (P<0.05). CCK-8 and scanning electron microscopy showed that PDLCs proliferated stably on the composite scaffold (P>0.05). Co-cultured with PDLCs, the PCL/COLI/nCZ group had more ALP staining and Alizarin Red staining areas than that of the PCL/COLI group (P<0.05), and there was no significant difference from the PCL/COLI/nHA group (P>0.05). Conclusions:The PCL/COLI/nCZ composite scaffold has excellent mechanical properties and biocompatibility, and has the potential for osteoinduction, which can be used for bone tissue engineering.

Objective:?To investigate the physicochemical properties and osteoinductive potential of magnesium phosphate scaffold with two different scale pores prepared by 3D printing technology and porogen leaching. Methods: Magnesium phosphate scaffolds (MgP) were prepared by indirect printing method. We used sodium chloride (particle size 25-50 μm) as porogen to fabricate microporous magnesium phosphate scaffolds (MgP-Na). The physical and chemical properties of magnesium phosphate scaffolds were examined by scanning electron microscopy, mercury porosimetry, general material testing machine and X-ray diffraction. CCK8 and DAPI staining were used to detect the proliferation and adhesion of human bone marrow mesenchymal stem cells (hBMSCs) seeded on scaffolds. Alizarin red staining and alkaline phosphatase quantitative assay were used to evaluate the ability of scaffolds to promote the osteogenic differentiation of hBMSCs. Results:MgP-Na scaffolds showed higher porosity, and the micropores significantly reduced the compressive strength of MgP-Na scaffolds (P＜0.05). Both scaffolds possessed excellent biocompatibility, and hBMSCs can adhere and proliferate on the scaffolds (P＜0.05). Moreover, the microporous structure was more conducive to cell adhesion. MgP-Na scaffolds played a more active role in calcium deposition and osteogenic differentiation of hBMSCs seeded on scaffolds (P＜0.05). Conclusions: Magnesium phosphate scaffold is expected to be a new material for bone defects repairing. The microporous structure of MgP-Na scaffold plays a more effective role in osteoinduction.

Objective:The aim was to investigate correlation between expression level of miR-299-5p and miR-127-3p in peripheral blood and nonsyndromic orofacial clefts (NSOC). Methods:Ten cleft lip with or without cleft palate (CL/P) patients, 10 cleft palate only (CPO) and 10 healthy children were selected from the First Affiliated Hospital of Harbin Medical University, using real-time RT-PCR（qRT-PCR）to detect the relative expression of miR-299-5p miR-127-3p in the peripheral blood from three groups. The receiver operating characteristic (ROC) curve and area under the curve (AUC) were applied to analyze the diagnostic efficacy of miR-299-5p and miR-127-3p for NSOC. Results:Compared with the healthy control group, the expression of miR-299-5p and miR-127-3p in CL/P group and CPO group was down-regulated, and the difference was statistically significant (P<0.05). The AUC of miR-299-5p and miR-127-3p in the diagnosis of NSOC were 0.770 and 0.810, respectively. The AUC of miR-299-5p and miR-127-3p in the combined diagnosis of NSOC was 0.915. Conclusions:The down-regulation of miR-299-5p and miR-127-3p in peripheral blood of children with NSOC may be related to the progression of NSOC, which may become a potential biomarker for the diagnosis of NSOC, and the combination of the two miRNAs is more effective.

Objective:To collect and analyze the whole genome transcription information of gingival tissue of patients with impacted maxillary canine, and lay a foundation for the gene theory of the etiology. Methods:Using resequecing Illumina hiseq2000 sequencing platform, the transcriptome information of gingival tissue of 5 adult patients (25-35 years old) with impacted maxillary canines in Zunyi area was collected by high-throughput sequencing. The expression level and difference of gene in gingival tissue were analyzed and calculated by FPKM method. Results:223530890 high quality sequencing data were obtained, and 19352 genes were indentified to be expressed in the gingival tissue, among which 99.63% genes’ FPKM values were lower than 1000. 15412 genes had alternative splicing (AS), and the aplicing times were 16167. Mutually exclusive exons were found to be the rarest AS type (6.72%). While skipped exons found to be the major type (48.82%). Using these transcriptome data, 386414 SNPs were called, among which transitions were the main type (76.31%). According to GO database, transcriptome SNPs were involved in a wide range of biological functions, and are especially enriched in biological processes and cell components; KEGG pathway analysis shows that the genes of SNPs are enriched in metabolic pathways, endocytosis, apoptosis, cell aging and other related pathways, and more genes are enriched in cancer, signaling pathways and immune system related pathways. Conclusions:Genome transcription information of gingival tissue of patients with impacted maxillary canine showed gingiva were closely related to the development of teeth.

Objective：To evaluate the effects of the abutment color and adhesive types on the color of restorations using ultra-thin porcelain laminate veneers. Methods：Six dental adhesives of two types were selected: 3 of Etch&rinse systems：Ivoclar Syntac? heliobond(SH), 3M AdperTM Single Bond 2(SB2)，Bisco One-Step Plus(OSP)，3 of Universal system: Ivoclar Tetric? N Bond Universal(TN)、, 3M Single Bond Universal(SBU), and Bisco ALL-Bond Univeral(ABU) . The specimens of ultra-thin porcelain laminate veneers with the slices of veneer, cement, adhesive and resin replacement（thickness of 0.3mm，0.1mm，0.1mm，and 2mm respectively）were prepared. Resin replacements of IPS Natural die from ND1 to ND4 were used to imitate different color of the abutments. The color differences of different ultra-thin porcelain laminate veneers were measured using Vita EasyShade spectrophotometer and the values of color differences (ΔE) were calculated. Results：The type of adhesives, the abutment colors and their interaction all showed a significant effect on the ΔE values of the specimens of ultra-thin porcelain laminate veneers (P<0.001). When the abutment color was the same, the color differences between OSP and ABU was not significant and there was a significant difference within groups of TN，SH, SBU ,and SB2 : TN > SH, SBU > SB2(P < 0.05). When the type of adhesives was the same, the abutment colors of TN and SBU groups showed a significant effect on the color differences of ultra-thin porcelain laminate veneers: ND2 > ND3 > ND1 > ND4 in TN group (P < 0.05); and ND1 > ND2 > ND3 > ND4 in SBU group (P < 0.05). Conclusions： Both the type of adhesives and the abutment color showed an effect on the color of ultra-thin porcelain laminate veneers. Therefore, dentists should pay attention to the color of abutment and select an optimal dental adhesive when bonding a ultra-thin porcelain laminate veneer in clinic.

Objective：Study on changes of airway before and after mandibular advancement orthoganic treatment in skeletal Ⅱ adult female. Methods：Forty skeletal Ⅱ adult female patients were divided into 2 groups: orthognathic group (20 patients) and orthodontic group (20 patients). CBCT was taken before and after treatment. The airway volume and cross-sectional area were measured before and after orthodontic treatment. SPSS 25.0 was used for analyzing the changes by t test. Results：After treatment, all the measurements in the orthodontic group were not significantly changed, while the total airway volume, the minimal cross sectional area and lower cross sectional area were significantly increased in orthognathic group. Conclusions：Mandibular advancement by sagittal splint ramus osteotomy can significantly increase the airway volume, minimal cross-sectional area and lower cross sectional area in skeletal Ⅱ adult female, improve respiratory function, while orthodontic treatmenthas no significant effect on airway in these skeletal Ⅱ adult female.

Objective：To measure the anatomical data of root surface of three-rooted PMFMs in adolescents with CBCT, to provide theoretical support and clinical guidance for the prognosis and treatment of periodontitis. Methods：175 CBCT images of three-rooted PMFMs were collected. The occurrence of teeth, the length of root, the length of root trunk, the degree of concavities of root trunk at the level of 2mm below CEJ, the width of root bifurcation and the degree of root concavities at the level of 2mm below apex of root bifurcation vault and the depth of concavities at different levels of mesial root surface were recorded and analyzed. Results：The length of mesial root、distal buccal root and distal lingual root were 13.06±1.26mm,11.85±1.30mm and 10.26±1.61mm respectively. Except for distal lingual root,the root length of male was longer than that of female(P<0.05).The length of distal root trunk (4.87±1.00) mm was the longest, and the length of buccal root trunk(2.56±0.74) mm was the shortest.The degree of root trunk concavities at the distal side was the smallest at the level of 2mm below CEJ.The width of buccal root bifurcation was the broadest at the level of 2mm below apex of root bifurcation vault , with an average width of 2.77±0.67mm.The depth and degree of concavities of the mesial root-surface were greater than those of the distal buccal one. From the level of 2mm to 6mm below apex of root bifurcation vault, the depth of the concavities of mesial root surface became shallow gradually.The above differences were statistically significant(P<0.05).Conclusions:The three-rooted PMFMs had long distal root trunk.Once the lesion extends to the root bifurcation,the treatment will be difficult and the prognosis will be poor. The mesial root was obviously depressed.When the lesion spreads here, the treatment can be entered from the buccal bifurcation.

Peroxisome proliferator-activated receptors（PPARs） are ligand-stimulating receptors in the nuclear receptor family, including three isotypes：PPARα、PPARβ／δ and PPARγ. PPARα has been implicated as a major regulator of lipid metabolism and inflammation, PPARβ／δ appears to be associated with mammalian skin, liver and bone regeneration, PPARγ has emerged as a key regulator of fat formation and glucose homeostasis. This review summarizes our current knowledge of PPARs agonists in processes associated with osteoporosis.