OBJECTIVETo study the expression of Runx2/Cbfa1 in the developing dentin and differentiating odontoblasts.

METHODSA postnatal mice teeth developing model was built histologically. Immunohistochemical technique was adopted to determine the expression of Runx2/Cbfa1 in the developing pulpo-dentinal complex in mice.

RESULTSRunx2/Cbfa1 was merely present in predentin in the exact and before the 11th day's postnatal stages. Meanwhile, it was positively located in odontoblasts and dental pulp cells in root region, but negatively in coral part after the 11th day's stages.

CONCLUSIONRunx2/Cbfa1 may play an important role in the deposing of tooth dentin and in the differentiating of odontoblasts and pulp cells.

Animals ; Cell Differentiation ; Core Binding Factor Alpha 1 Subunit ; Dental Pulp ; Dentin ; Mice ; Odontoblasts ; Tooth

OBJECTIVECleidocranial dysplasia (CCD) is a rare skeletal disease with autosomal dominant inheritance associated with mutation in RUNX 2. The authors report a Chinese girl with CCD in whom the mutation in RUNX 2 was identified.

METHODSClinical diagnosis was based on physical examination, radiological findings, and biochemical tests. For mutation detection, genomic DNA was extracted from peripheral blood using standard method. All 7 coding exons of RUNX 2 and their flanking intronic sequences were amplified by polymerase chain reaction (PCR), and the PCR products were then subjected to automatic DNA sequencing.

RESULTSThe affected girl showed typical clinical manifestations of CCD, including patent fontanelles, absent clavicles, short stature and dental anomalies. Direct sequencing of PCR-amplified fragments revealed a recurrent missense mutation, R190W (568 C > T), in RUNX 2. The mutation was further confirmed by Hae III restriction analysis.

CONCLUSIONA Chinese case of CCD was confirmed and the disease-causing mutation was linked to a recurrent point mutation in RUNX 2.

Cleidocranial Dysplasia ; genetics ; Core Binding Factor Alpha 1 Subunit ; genetics ; Female ; Humans ; Mutation

OBJECTIVETo evaluate the effects of mechanical stress on the formation and expression of core binding factor alpha1 (Cbfalpha1) in MG-63 cells cultured on titanium in vitro.

METHODSMG-63 cells cultured on the titanium were subjected to a centrifugal force (2.205 N) 15 min per 4 hours and collected after 4, 8 and 12 hours. The formation and expression of Cbfalpha1 were examined by immunofluorescence staining and reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSBoth the cells with or without centrifugal force created the fluorescence in the nucleus and the immunofluorescence intensity of Cbfalpha1 in MG-63 cells with centrifugal force were higher than those without centrifugal force (P<0.05). Meanwhile, both the cells with or without centrifugal force expressed the mRNA of Cbfalpha1 and the relative mRNA level of Cbfalpha1 in MG-63 cells with centrifugal force were higher than those without centrifugal force, and the differences were great significant (P<0.05).

CONCLUSIONMechanical stress are beneficial to the formation and expression of Cbfalpha1.

Core Binding Factor Alpha 1 Subunit ; In Vitro Techniques ; RNA, Messenger ; Stress, Mechanical

OBJECTIVE: To explore the genetic basis for a Chinese patient featuring cleidocranial dysplasia(CCD). METHODS: Genomic DNA was extracted from peripheral blood samples of the patient and his parents. Whole exome sequencing (WES) was carried out for the patient, and suspected variant was verified by Sanger sequencing. RESULTS: WES has identified a missense c.460G>T (p.Val154Phe) (GRCh37/hg19) variant of the RUNX2 gene. The variant was located in the Runt domain, a highly conserved region (PM1); it was not present in either the Genome Aggregation Database or the 1000 Genomes Project (PM2), and was predicted to have a deleterious effect on the gene product by multiple in silico prediction tools (PP3); the clinical phenotype of the patient was highly consistent with that of cleidocranial dysplasia (PP4). Furthermore, the variant was unreported in medical literature and was absent in both parents (PS2). Based on the American College of Medical Genetics and Genomics guidelines, the c.460 G>T variant of RUNX2 gene was predicted to be pathogenic (PS2+PM1+PM2+PP3+PP4). CONCLUSION The c.460G>T (p.Val154Phe) variant of the RUNX2 gene probably underlay the clinical phenotype in the patient. Above finding has enabled accurate diagnosis and expanded the spectrum of RUNX2 variants.
China ; Cleidocranial Dysplasia/genetics* ; Core Binding Factor Alpha 1 Subunit/genetics* ; Humans ; Mutation ; Whole Exome Sequencing

Cleidocranial dysplasia is a rare autosomal dominant hereditary disease characterized by abnormal skeletal and dental development. In this work, a case of cleidocranial dysplasia is reported, and a new frameshift mutation is confirmed by gene detection.
Cleidocranial Dysplasia ; Core Binding Factor Alpha 1 Subunit ; Diagnostic Tests, Routine ; Humans ; Mutation

OBJECTIVE: To detect the genetic variant of a child with cleidocranial dysplasia (CCD) and to find out the causation of the illness. METHODS: Gene variant was identified by the second generation targeted sequencing and Sanger sequencing. RESULTS: The gene sequencing revealed that the RUNX2 gene had c.196C>T(p.Glu66*) nonsense variant, which was predicted to be a pathogenic variant according to the ACMG guidelines(PVS1+PS2). CONCLUSION The variant of c.196C > T in the RUNX2 gene may be the cause of the child with CCD, and the novel variant enriches the RUNX2 gene variant spectrum.
Asians/genetics* ; Child ; China ; Cleidocranial Dysplasia/genetics* ; Core Binding Factor Alpha 1 Subunit/genetics* ; Humans ; Mutation

OBJECTIVE: To study the clinical features and prognosis of core binding factor acute myeloid leukemia (CBF-AML) in children. METHODS: A retrospective analysis was performed from the chart review data of children who were newly diagnosed with CBF-AML in the Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences, from August 2009 to November 2015. According to the type of fusion gene, the children were divided into CBFB-MYH11 and AML1-ETO groups. Clinical features and prognosis were analyzed and compared between the two groups. RESULTS: A total of 91 children with CBF-AML were enrolled in this study, among whom there were 74 (81%) in the AML1-ETO group and 17 (19%) in the CBFB-MYH11 group. Additional chromosomal abnormalities were observed in 38 children (42%), and deletion of sex chromosome was the most common abnormality and was observed in 28 children (31%). After the first course of induction treatment, the complete remission rate was 97% (88/91), the recurrence rate was 29% (26/91), the 5-year event-free survival (EFS) rate was 65%±6%, and the 5-year overall survival (OS) rate was 75%±5%. There were no significant differences between the AML1-ETO and CBFB-MYH11 groups in 5-year EFS rate (62%±7% vs 77%±11%, P>0.05) or 5-year OS rate (72%±6% vs 88%±9%, P>0.05). CONCLUSIONS AML1-ETO is the main type of fusion gene in children with CBF-AML, and deletion of sex chromosome is the most common type of additional chromosomal abnormalities. Children with CBF-AML often have a good prognosis, and the children with AML1-ETO have a similar prognosis to those with CBFB-MYH11.
Child ; Core Binding Factor Alpha 2 Subunit ; Core Binding Factors ; Humans ; Leukemia, Myeloid, Acute ; Oncogene Proteins, Fusion ; Prognosis ; RUNX1 Translocation Partner 1 Protein ; Retrospective Studies

OBJECTIVEThis study investigated the rapid response of osteoblasts, which were derived from low-magnitude high-frequency vibration (LMHFV). Refractory period-derived memory response was also observed.

METHODSMC3T3-E1 cells were incubated and received LMHFV stimulation (0.49 g, 40 Hz) for 30 min. After application of LMHFV, mRNA levels of earlier osteogenic differentiation markers Runt-related transcription factor 2 (Runx2), collagen typeⅠ(Col-Ⅰ), and alkaline phosphatase (ALP) were immediately detected by real-time fluorescence quantitative polymerase chain reaction in the absence or presence of antioxidant. Simultaneously, concentrations of mitochondrial reactive oxygen species (ROS) and average mitochondrial length were also measured.

RESULTSOsteoblasts in the vibration group showed decreased gene expressions of Runx2, Col-Ⅰ, and ALP (P<0.01) and increased levels of mitochondrial ROS (P<0.01) and shortened mitochondria (P<0.01), whereas antioxidant treatment resulted in recovery from changes in the above indicators (P<0.01).

CONCLUSIONSLMHFV can downregulate mRNA levels of early osteogenic differentiation markers, promote ROS generation, and mitochondrial fission. 
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Alkaline Phosphatase ; Animals ; Cell Line ; Core Binding Factor Alpha 1 Subunit ; Down-Regulation ; Mice ; Osteoblasts ; Osteogenesis ; Vibration

OBJECTIVETo evaluate the effect of concentrated growth factor extract (CGFe) on the proliferation and differentiation of MC3T3-E1 osteoblasts attached to sandblasted and acid etched titanium surfaces.

METHODSTrials were divided into experimental and control groups. The experimental group used a-MEM that contained CGFe (10% FBS), whereas the control group only used a-MEM (10% FBS). MTT assay was employed to detect the number of osteoblasts on the first, third, and fifth days. Alkaline phosphatase (ALP) activity and scanning electron microscope (SEM) were used to detect the osteoblast differentiations on the third and fifth days and to observe the osteoblast extensions on titanium surfaces for 12 h, respectively. Meanwhile, the levels of the osteogenetic biomarkers Runt-related transcription factor-2 (Runx2) and Osterix (Osx) on the third and seventh days were quantified via real-time polymerase chain reaction (PCR).

RESULTSMTT assay indicated that on the first, third, and fifth days, the absorbance in the experimental group significantly increased than that in the control group (P < 0.05). ALP activity: on the third and fifth days, the absorbance of the experimental group was significantly higher than that of the control group (P < 0.05). SEM: at 12 h, the extension of the experimental group cells was larger than that of the control group. Real-time PCR: given the standardization in the group, the gene expression level of the control group on the third day was 1, and the Runx2 and Osx gene expressions in the experimental group were larger than those of the con- trol group.

CONCLUSIONCGFe can efficiently stimulate the proliferation, differentiation and extension of MC3T3-E1 cells.

Cell Differentiation ; Cell Line ; Cell Proliferation ; Core Binding Factor Alpha 1 Subunit ; Intercellular Signaling Peptides and Proteins ; Osteoblasts ; Osteogenesis ; Titanium

OBJECTIVETo explicit whether the expression of the mineral-related proteins is regulated by cbfa1 in human dental papilla cells.

METHODSHuman dental papilla cells were cultured in vitro and transfected with pcDNA3-cbfa1 recombinant plasmids. After selected with G418 sulfate, a cell clone named PC-3, which could stably express the cbfa1 mRNA and protein, was proved by PCR and western blot. Then the amount of ALP and OC and the expression of OPN, BSP, ON, DMP1, DSP and DSPP were detected by immunohistochemistry, Western blot and PCR methods.

RESULTSWe established the human dental papilla cells model PC-3 which could stably express the cbfa1 mRNA and protein. Compared with normal cells, a lot of mineral-related proteins such as ALP, OC, OPN, BSP, ON, DMP1 were upregulated in PC-3 cells.

CONCLUSIONSIn human dental papilla cells, cbfa1 can induce the expression of most mineral-related genes and proteins. It may implicate that cbfa1 must play a key role during tooth development and mineralization.

Alkaline Phosphatase ; biosynthesis ; Cells, Cultured ; Core Binding Factor Alpha 1 Subunit ; physiology ; Dental Papilla ; cytology ; metabolism ; Humans ; Osteocalcin ; biosynthesis