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Wang J.-H.,Chongqing Key Laboratory for Oral Diseases and Biomedical science | Wang J.-H.,Chongqing Medical University | Liu Y.-Z.,Chongqing Medical University | Liu Y.-Z.,Key Laboratory for Biochemistry and Molecular Pharmacology of Chongqing | And 16 more authors.
Bone | Year: 2013

Mesenchymal stem cells (MSCs) can self-renew and differentiate into osteogenic, chondrogenic, adipogenic and myogenic lineages. It's reported that bone morphogenetic protein 9 (BMP9) is one of the most potent osteogenic BMPs to initiate the commitment of MSCs to osteoblast lineage. Cyclooxygenase-2 (COX-2) is critical for bone fracture healing and osteogenic differentiation in MSCs. However, the relationship between COX-2 and BMP9 in osteogenesis remains unknown. Herein, we investigate the role of COX-2 in BMP9-induced osteogenesis in MSCs. We demonstrate that COX-2 is up-regulated as a target of BMP9 in MSCs. Both COX-2 inhibitor (NS-398) and COX-2 knockdown siRNAs can effectively decrease alkaline phosphatase (ALP) activities induced by BMP9 in MSCs. NS-398 also down-regulates BMP9-induced expression of osteopontin and osteocalcin, so does the matrix mineralization. The in vivo studies indicate that knockdown of COX-2 attenuates BMP9-induced ectopic bone formation. In perinatal limb culture assay, NS-398 is shown to reduce the hypertropic chondrocyte zone and ossification induced by BMP9. Mechanistically, knockdown of COX-2 significantly inhibits the BMP9 up-regulated expression of Runx2 and Dlx-5 in MSCs, which can be rescued by exogenous expression of COX-2. Furthermore, knockdown of COX-2 apparently reduces BMP9 induced BMPR-Smad reporter activity, the phosphorylation of Smad1/5/8, and the expression of Smad6 and Smad7 in MSCs. NS-398 blocks the expression of BMP9 mediated by BMP9 recombinant adenovirus. Taken together, our findings suggest that COX-2 plays an important role in BMP9 induced osteogenic differentiation in MSCs; BMP9 and COX-2 may form an important regulatory loop to orchestrate the osteogenic differentiation in MSCs. © 2013 Elsevier Inc. Source

He S.-L.,Chongqing Key Laboratory for Oral Diseases and Biomedical science | He S.-L.,Chongqing Medical University | Wang J.-H.,Chongqing Key Laboratory for Oral Diseases and Biomedical science | Wang J.-H.,Chongqing Medical University | And 2 more authors.
Quality of Life Research | Year: 2013

Objectives The aim of this study was to validate the Chinese version of the Summated Xerostomia Inventory (SXI). Methods The English SXI was translated into Chinese, cross-cultural adaptation and pilot tested. The final Chinese version of SXI was tested in a consecutive sample of 212 patients with xerostomia. The reliability of the Chinese version of SXI was determined through internal consistency and test-retest methods. The construct validity of SXI was analysed by content validity, construct validity, and convergent validity. Results Cronbach's alpha value for the SXI score was 0.798, and the test-retest intraclass correlation coefficient value for the SXI score was 0.837. Construct validity was proved by the presence of one-factor structure that accounted for 57.68 % of the variance and fitted well into the model. The correlation between the total score of the SXI and the global oral health question was 0.75, indicating very good correlation (P \ 0.01). Conclusion This study provided preliminary evidence concerning validity and reliability of the Chinese version of the SXI. The results provide initial evidence that the SXI may be a useful tool for the mainland Chinese xerostomia patients for both clinical and epidemiologic researches. © Springer Science+Business Media Dordrecht 2013. Source

Pang L.,Chongqing Medical University | Pang L.,Chongqing Key Laboratory for Oral Diseases and Biomedical science | Zhao X.,Chongqing University | Liu W.,Chongqing Medical University | And 5 more authors.
Nutrients | Year: 2015

Bear bile was used as a traditional medicine or tonic in East Asia, and ursodeoxycholic acid (UDCA) is the most important compound in bear bile. Further, synthetic UDCA is also used in modern medicine and nutrition; therefore, its further functional effects warrant research, in vitro methods could be used for the fundamental research of its anticancer effects. In this study, the apoptotic effects of UDCA in human oral squamous carcinoma HSC-3 cells through the activation of caspases were observed by the experimental methods of MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide) assay, DAPI (4’,6-diamidino-2-phenylindole) staining, flow cytometry analysis, RT-PCR (reverse transcription-polymerase chain reaction) assay and Western blot assay after HSC-3 cells were treated by different concentrations of UDCA. With 0 to 400 μg/mL UDCA treatment, UDCA had strong growth inhibitory effects in HSC-3 cells, but had almost no effect in HOK normal oral cells. At concentrations of 100, 200 and 400 μg/mL, UDCA could induce apoptosis compared to untreated control HSC-3 cells. Treatment of 400 μg/mL UDCA could induce more apoptotic cancer cells than 100 and 200 μg/mL treatment; the sub-G1 DNA content of 400 μg/mL UDCA treated cancer cells was 41.3% versus 10.6% (100 μg/mL) and 22.4% (200 μg/mL). After different concentrations of UDCA treatment, the mRNA and protein expressions of caspase-3, caspase-8, caspase-9, Bax, Fas/FasL (Fas ligand), TRAIL (TNF-related apoptosis-inducing ligand), DR4 (death receptor 4) and DR5 (death receptor 5) were increased in HSC-3 cells, and mRNA and protein expressions of Bcl-2 (B-cell lymphoma 2), Bcl-xL (B-cell lymphoma-extra large), XIAP (X-linked inhibitor of apoptosis protein), cIAP-1 (cellular inhibitor of apoptosis 1), cIAP-2 (cellular inhibitor of apoptosis 2) and survival were decreased. Meanwhile, at the highest concentration of 400 μg/mL, caspase-3, caspase-8, caspase-9, Bax, Fas/FasL, TRAIL, DR4, DR5, and IκB-α expression levels were the highest, and Bcl-2, Bcl-xL, XIAP, cIAP-1, cIAP-2, survival, and NF-κB expression levels were the lowest. These results proved that UDCA could induce apoptosis of HSC-3 cancer cells through caspase activation, and the higher concentration of UDCA had stronger effects in vitro. UDCA might be a good nutrient for oral cancer prevention. © 2015 by the authors; licensee MDPI, Basel, Switzerland. Source

Li Y.,Chongqing Medical University | Li Y.,Chongqing Key Laboratory for Oral Diseases and Biomedical science | Zhou Z.,Chongqing Medical University | Zhou Z.,Chongqing Key Laboratory for Oral Diseases and Biomedical science
Journal of Pure and Applied Microbiology | Year: 2014

To construct LuxS deletion mutant of Streptococcus mutans, and study the effect of luxS mutation on the biofilm formation of streptococcus mutans under various conditions, and find out the differences between luxS mutant strain and streptococcus mutans. Long flanking homology polymerase chain reaction(LFH-PCR) was introduced to generate a gene disruption construct consisting of Emr cassette with long flanking homology regions to the target gene. The electroporation competence of Streptococcus mutans was then transformed with this PCR product. Then positive transformants were counted on selective agar which containing erythromycin and identified by PCR. The streptococcus mutans-luxS mutant and the standard strain were grown in three different conditions(BHI, 2% glucose-BHI, 2% saccharobiose-BHI), and the ability of S.mutans and LuxS mutant biofilm formation was examined in 24 h by scanning electron microscopy (SEM). Identification by PCR and sequencing confirmed the validity of the LuxS deletion mutant of Streptococcus mutans. Compared with S.mutans ,the LuxS mutant maintained with 2% sucrose displayed an apparent defect in biofilm formation. Conclusions: The successful construction of the LuxS deletion mutant, and the ability of sucrose-dependent biofilm formation will be down-regulated in Streptococcus mutans after LuxS gene was knocked out. Source

Deng J.-S.,Chongqing Medical University | Deng J.-S.,Chongqing Key Laboratory for Oral Diseases and Biomedical science | Qin P.,Chongqing Medical University | Li X.-X.,Chongqing Medical University | And 2 more authors.
Human Immunology | Year: 2013

The aim of this study was to perform a meta-analysis to evaluated the association between interleukin-1β (IL-1β) C(3953/4)T polymorphism and chronic periodontitis (CP). Systematic searches of electronic databases and hand searching of references were performed, including PubMed, Embase and Web of Science. The pooled odds ratios (ORs) with 95% confidence intervals (95%CIs) were calculated. Publication bias was tested by Begg's funnel plot and Egger's regression test. Sensitivity analysis was conducted by limiting the meta-analysis studies conforming to Hardy-Weinberg equilibrium (HWE) or high quality (score. ≥7). Data analyses were carried out by Stata 11.0. There were significant associations between IL-1β C(3953/4)T polymorphism and CP (for T allele vs. C allele: OR. =1.30, 95%CI. =1.05-1.60, p=0.02; for T/T vs. C/C: OR. =1.66, 95%CI. =1.12-2.45, p=0.01; for C/T. +. T/T vs. C/C: OR. =1.28, 95%CI. =0.99-1.65; and for T/T vs. C/T+C/C: OR. =1.62, 95%CI. =1.15-2.29, p=0.006). When stratified by ethnicity, statistically significantly elevated risk was found for Caucasians, but not for Asians. When stratified by study design, evidences of significant association was observed between IL-1β C(3953/4)T polymorphism and CP in both population-based studies and hospital-based studies. This meta-analysis indicates that there is strong evidence for association between IL-1β C(3953/4)T polymorphism and CP. © 2012 American Society for Histocompatibility and Immunogenetics. Source

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