Objective: To construct mcpr1prokaryoti c expression vector and to express MCPR1 protein.Methods:PCR was used to obtain coding region of mcpr1. Construction of a high-level fusion protein expression vector pGEX-4T-mcpr1 was conducted by inserting the fra gment of coding region of mcpr1into a fusion protein expression vector pGEX -4T-1. Then the recombinant plasmid was transferred into E. colito prepar e the MCPR1/GST fusion protein. DNA sequencing and endonucleases digesting were used to check the coding region. Results:pGEX-4T- mcpr1 wa s constructed successfully and the coding region was inserted into the vector co rrectly. A new protein band of 36 000 was observed by SDS-PAGE analysis after i nduction by IPTG. The 36 000 protein amounted to 39 percent of the total prote in and existed mostly in precipitation of broken bacteria. Conclusion: MCPR1 protein can be expressed in E. coliexpression system and purif ied initially.

Objective: To analyze mutations of RUNX2 gene in a Chinese family with CCD. Methods: The proband and her parents were investigated in the present study. Radiological examination regarding osseous malformations was carried out over the entire body. Genomic DNA was extracted from whole blood, and the RUNX2 gene was amplified by PCR from genomic DNA. 100 healthy people were also included. DNA sequences were analyzed by using BLASTN (BLAST nucleotide) program. Results: Both the proband and her mother have typical CCD clinical characteristics, different from her healthy father. After BLASTN analysis, one novel mutation was identified in the proband and her mother, a heterozygous A to G transition mutation at nucleotide 478 in exon 2, which converted asparagines to aspartic acid at codon 160 (478 A>G,N160D). Conclusion: The N160D mutation is identified as a novel heterozygous mutation, which supplements the data of RUNX2 gene mutation research.

Objective To explore the splenic macrophages phenotype and secretory function of mice with periodontitis, so as to explore effects of periodontitis on macrophages. Methods 22 mice were randomly divided into periodental ligation group (group P10d ) and pseudo periodental ligation control group (group C), with 11 mice in each group. The experimental periodental ligation on mice lasted for 10 days before they were sacrificed. Flow cytometry was applied to detect the expression of M1 and M2 in mononuclear macrophages. Real-time PCR was applied to detect the relative expression of pro-inflammatory cytokines IL-1β and anti inflammatory cytokines IL-10. Results Compared with the control group C, the proportion of M1 macrophages in the periodontitis group decreased, and the ratio of M1/M2 was also decreased significantly, and IL-1β mRNA also down-regulated. Conclusions Chronic periodontal infection could down regulate the proportion of M1 macrophages , decrease ratio of M1/M2 and the expression of inflammatory cytokines IL-1βmRNA.

Objective:To study the expression of B-1a cells in mice with obesity and periodontal infection.Methods:Mouse models of diet induced obesity combined with experimental periodontitis were established,the expression of CD5 protein,anti-collagenⅠ antibody(anti-Col-Ⅰantibody) and IL-10 protein was examined in mouse jaw bone and spleen by immunohistochemistry and Western blot;The mRNA expression of CD5,anti-Col-Ⅰantibody and IL-10 in mouse jaw bone was detected by real time quantitative PCR.Results:The mRNA and protein expressions of CD5 and IL-10 and anti-Col-Ⅰantibody in jaw bone in periodontitis group were significantly higher than those in control group(P<0.001).The protein expressions of CD5 and IL-10 and anti-Col-Ⅰantibody in spleen in obesity group were significantly higher than those in standard group(P<0.05).The protein expression of anti-Col-Ⅰantibody in spleen in standard accompanying periodontal ligature group was significantly higher than that in standard without periodontal ligature group(P<0.05).Conclusion:B-1a cells are activated in the early stage of obesity and periodontal inflammation with a certain pathological significance and without interation between the two inflammatory states in the pathological mechanism.

BACKGROUND:Hard palate mucosa serves as a main donor material in periodontal plastic surgery and its thickness is crucial for the surgical outcomes. OBJECTIVE: To analyze the thickness of hard palate mucosa in Han population, and analyze the consistency between cone-beam CT image analysis and trans-gingival probing method. METHODS: A total of 30 Han volunteers (300 teeth) were recruited, and the thickness of hard palate mucosa was measured using cone-beam CT image analysis or trans-gingival probing method, to analyze their consistency. RESULTS AND CONCLUSION: The two methods showed a higher consistency in the thickness of hard palate mucosa at the cuspid, first and second premolars as well as first and second molars. The thickness of the hard palate mucosa related to the distance from the gingival margin and tooth position, the thickness from the canine region to the second premolar region thickening gradually, and became the thickest at the second molar, and the thinnest at the cuspid. This study for the first time analyzed the thickness of hard palate mucosa in Chinese Han population, and confirmed there is a high consistency between cone-beam CT image analysis and trans-gingival probing method.

Objective:To observe the temporal and spatial expression and function of mcpr1 gene during murine tooth germ development.Methods:The expression of MCPR1 at different stages of mouse tooth germ were detected by immunohistochemical staining.Results:MCPR1 expression was detected at all stages of tooth germ, but the distribution patterns at various stages were different. It indicated that the temporal and spatial expression pattern of MCPR1 during murine tooth germ development was specific.Conclusion:mcpr1 might play an important role in modulating the differentiation and mature of enamel organ.

Objective: To investigate the relationship between TIMP-1 gene polymorphisms and susceptibility of chronic periodontitis(CP) in Han Cantonese. Methods: Buccal swabs from 70 CP patients and 74 periodontal healthy controls were collected. DNA was extracted from these buccal swabs by using Chelex-100. TIMP-1 +372T/C (rs4898)、TIMP-1+533C/T (rs1062849) polymorphisms were tested by polymerase chain reaction - restriction fragment length polymorphism (PCR-RFLP). Allele distribution and genotypes frequencies in the patients and controls were analyzed. Results: Frequency variance of allele T and C at site of TIMP-1 +372 in patients and controls showed no statistical difference. TIMP-1+533C/T polymorphism of TIMP-1 hadn't been found in the present study. Conclusion: There is no relationship between TIMP-1 +372T/C polymorphism and susceptibility of chronic periodontitis, and TIMP-1 +533C/T polymorphism doesn' t exist among Han Cantonese.

Objective:To evaluate the effects of periodontitis on the expression of apoptosis-related proteins in pancreas of rats with Type 2 Diabetes Mellitus.Methods:Spontaneously type 2 diabetic OLETF rats were randomly divided into 2 groups:diabetes with or without periodontitis(diabetes group and combination group).LETO rats with the same germline and the same age but having normal glucose tolerance were randomly divided into control group and periodontitis group.20 weeks after periodontitis were established,all the rats were sacrificed and the pancreas were pathologically examined by HE staining.The expression of Bax,Bcl-2 and Caspase-3 in the pancreas islet were detected by immunohistochemistry staining and semi-quantitative analysis.Results:The expression of Bax, Bcl-2 and Caspase-3 in the pancreas islet was no significant difference between control and periodontitis groups(P=0.324,P=0.091,P=0.852).Compared with diabetes group,the expression of Bax and Caspase-3 in combination group showed a significant increase(P=0.000,P=0.000),and the expression of Bcl-2 was significantly decreased(P=0.022).Conclusion:Under healthy conditions,periodontitis has no effect on the expression of apoptosis-related proteins in rat pancreas islet.However,in rats with diabe-tes,periodontitis may affect the expression of apoptosis-related proteins in pancreas islet.

OBJECTIVETo explore the role of platelet-activating factor receptor (PAFR) in adhesion and invasion of phospho- rylcholine (PC)-positive Aggregatibacter actinomycetemcomitans in cultured human umbilical vein endothelial cells (HUVEC).

METHDOSCultured HUVECs were pretreated with the PAFR antagonist CV3988 or anti-human PAFR monoclonal antibody for 30 min before infection with PC-positive or -negative A. actinomycetemcomitans strains. The bacterial adhesion and invasion and cytotoxicity in the cells were examined using MTT assay.

RESULTSPretreatment with PAFR antagonists at 100, 200 and 500 nmol/L significantly reduced the adhesion rate (36.29∓3.52)%, (19.04∓3.35)% and (7.69∓3.19%), respectively] and invasion rate [(12.12∓1.58)%, (7.08∓0.29)% and (2.60∓2.26)%, respectively] of PC-positive A.actinomycetemcomitans in HUVECs. Similarly, pretreatment with anti-PAFR antibody also significantly reduced A.actinomycetemcomitans adhesion and invasion in HUVECs [(50.05∓5.28)% and (39.09∓6.50)%, respectively]. Pretreatment with PAFR antagonist (200 and 500 nmol/L) and anti-PAFR antibody (25 µg/mL) significantly increased the viability of HUVECs incubated with PC-positive A.actinomycetemcomitans from (25.39∓9.33)% to (91.12∓3.14)%, (94.12∓2.15)% and (65.5∓1.87)%, respectively, but such pretreatments did not increase the viability of cells incubated with PC-negative A.actinomycetemcomitans.

CONCLUSIONSPAFR plays an important role in the adhesion, invasion, and cytotoxicity of PC-positive A.actinomycetemcomitans in cultured HUVECs.

Aggregatibacter actinomycetemcomitans ; pathogenicity ; Bacterial Adhesion ; Cells, Cultured ; Human Umbilical Vein Endothelial Cells ; microbiology ; Humans ; Platelet Membrane Glycoproteins ; metabolism ; Receptors, G-Protein-Coupled ; metabolism