Effects of Hybrid Coat on shear bond strength of five cements: an in vitro study

PURPOSE: To evaluate the sealing performance of Hybrid Coat and its influence on the shear bond strength of five dentin surface cements. MATERIALS AND METHODS: Six premolars were pretreated to expose the dentin surface prior to the application of Hybrid Coat. The microscopic characteristics of the dentinal surfaces were examined with scanning electron microscopy (SEM). Then, 40 premolars were sectioned longitudinally, and 80 semi-sections were divided into a control group (untreated) and a study group (treated by Hybrid Coat). Alloy restoration was bonded to the teeth specimen using five different cements. Shear bond strength was measured by the universal testing machine. The fracture patterns and the adhesive interface were observed using astereomicroscope. RESULTS: SEM revealed that the lumens of dentinal tubules were completely occluded by Hybrid Coat. The Hybrid Coat significantly improved the shear bond strength of resin-modified glass ionomer cement (RMGIC) and resin cement (RC) but weakened the performance of zinc phosphate cement (ZPC), zinc polycarboxylate cement (ZPCC) and glass ionomer cement (GIC). CONCLUSION: Hybrid Coat is an effective dentinal tubule sealant, and therefore its combined use with resin or resin-modified glass ionomer cements can be applied for the prostheses attachment purpose.

Expression of NF-κB and osteopontin of synovial fluid of patients with knee osteoarthritis

OBJECTIVE@#To explore the significance of osteopontin and nuclear factor κB (NF-κB) expression in patients with knee osteoarthritis.@*METHODS@#RT-PCR and enzyme-linked immunosorbent assay were used to measure the Osteopontin (OPN) and NF-κB concentration of knee joint synovial fluid of patients with knee osteoarthritis and trauma fractures, and analyze the relationship between the expressiones of them.@*RESULTS@#OPN and NF-κB expression at the mRNA and protein levels of patients with knee osteoarthritis were significantly higher than the control group. the result showed statistical significance (P<0.05). There was a positive correlation between the OPN levels in synovial fluid of patients with knee osteoarthritis and NF-κB expression levels (P<0.05).@*CONCLUSIONS@#The high expression of OPN and NF-κB are closely related to occurrence and development of knee osteoarthritis.

Construction of WISP3 gene's mutants in SEDT-PA and their expression in COS-7 cells / 中南大学学报(医学版)

OBJECTIVE@#To construct two types of Wnt-inducible secreted protein 3(WISP3) gene's mutants(1000T/C,840delT) found in spondyloepiphyseal dysplasia tarda with progressive anthopathy (SEDT-PA) patients, and to observe their expression in COS-7 cells.@*METHODS@#Full-length cDNA of wild type WISP3 gene(WT-WISP3) was amplified from human chondrocytes by RT-PCR, and site-directed mutagenesis was used to obtain full-length cDNAs of the mutated WISP3 genes(MUT1000T/C and MUT840delT). The recombined plasmids WT-WISP3/pcDNA3.1(+), MUT1000T/C/pcDNA3.1(+) and MUT840delT/pcDNA3.1(+) were transfected transiently into COS-7 cells by liposome-mediated method, and pcDNA3.1(+) vector was used as a control. The total RNA and protein of the transfected COS-7 cells were extracted after 48 hours of transfection. The expression of WISP3 gene in the transfected COS-7 cells was detected by semi-quantitative RT-PCR and Western blot.@*RESULTS@#By restriction endonuclease analysis and sequencing, the sequence of MUT1000T/C and MUT840delT were consistent with that mutated in SEDT-PA, and the open reading frames matched with the vector sequence. Semi-quantitative RT-PCR and Western blot showed that the recombined plasmids were highly expressed in COS-7 cells.@*CONCLUSION@#WISP3 gene's mutants of SEDT-PA are successfully constructed by genetic recombination, and expressed in COS-7 cells, which lays the foundation for the further study on its molecular functions in SEDT-PA.

Apoptosis of mesenchymal cell line MBA-1 induced by core binding factor alpha 1 / 中南大学学报(医学版)

OBJECTIVE@#To investigate the effect of two core binding factors alpha 1 (Cbfa1) isfroms (Cbfa1/P56 and Cbfa1/P57) on the apoptosis of mesenchymal cell line MBA-1.@*METHODS@#The two Cbfal isfroms were transiently transfected into MBA-1 cells, then the changes of apoptosis rate were observed by flow cytometer. The protein expressions of Cbfa1, Bax, Bcl-2, caspase-3, and caspase-9, cytochrome-C and TNF-alpha were determined by Western immunoblot.@*RESULTS@#After the transient transfection with the two isforms of Cbfa1, MBA-1, the cells apoptotic rates increased, and the ratio of Bax/Bcl-2, the expressions of cytochrome-C, caspase-3, caspase-9, and TNF-alpha were significantly increased.@*CONCLUSION@#Cbfa1 can promote the apoptosis in mesenchymal cell line MBA-1. Bax/Bcl-2, cytochrome-C, caspase-9, caspase-3, and TNF-alpha are also involved in the apoptosis pathway.

Effect of growth inhibition of the secretory protein SPLUNC1 on Pseudomonas aeruginosa / 中南大学学报(医学版)

OBJECTIVE@#To express the recombinant SPLUNC1 protein in HNE1 cells and to study its function of bactericidal and binding to lipopolysaccharide (LPS).@*METHODS@#Full length of SPLUNC1 gene was cloned into pCMV-tag4A vector and stably transfected into HNE1 cell lines, the supernatant of cell cultures was collected. After being treated with the supernatant, the Pseudomonas aeruginosa was seeded to LB soft agar plate, and the bacteria clones were counted and analyzed. For in vitro LPS binding assay, LPS was coated to 96-well plates. We incubated in the plate with SPLUNC1 protein, and detected the binded SPLUNC1 protein by ELISA. Incubating the FITC-LPS with the SPLUNC1 stably transfected or control cells, the intracellular intensity of fluorescence was observed under the fluorescence microscope.@*RESULTS@#SPLUNC1 inhibited the bacteria clone formation obviously. Although the binding efficiency of LPS and SPLUNC1 in vitro was very low, more FITC-LPS entered into the SPLUNC1 stably transfected cells.@*CONCLUSION@#SPLUNC1 can inhibit the growth of Pseudomonas aeruginosa and bind LPS, and play an important defensive role in innate immunity of the upper airway.

Preliminary biomarker related to nasopharyngeal carcinoma filtered from the whole genome expression profiling involved in microdissection nasopharyngeal tissues / 中南大学学报(医学版)

OBJECTIVE@#To filter biomarkers of nasopharyngeal carcinoma (NPC) by constructing the homogenesis tissue gene expression profiling with the whole human genome GeneChip.@*METHODS@#The epithelium cells of the homogenesis NPC and the pure nasopharyngeal normal tissues microdissected from nasopharyngeal biopsy which was preserved in the RNAlater were used to isolate RNA and then to harvest the aRNA through in vitro transcription, and aRNA prober was labled to hybridize to HG-U133. plus 2.0, so the expression profiling of each homogenesis tissue could be constructed.@*RESULTS@#Some candidate biomarker genes related to the tumorigenesis of NPC had been filtered by comparing the expression profiling of NPC samples with the expression profiling of normal nasopharyngeal epithelia samples. Any genes regarding the metastasis of NPC might have been selected by comparing the expression profiling of no-metastasis samples with those of the metastasis samples.@*CONCLUSION@#Using the whole genome GeneChip to construct the expression profiling for the microdissected homogenesis tissue is effective to filter the candidate biomarker genes.

Studies on the relationship between D6S1581, a high frequency allele imbalance locus, and genetic susceptibility to nasopharyngeal carcinoma / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the relationship of nasopharyngeal carcinoma (NPC) with the high frequency allele imbalance locus D6S1581, and the NPC associated gene FBXO30 which is located near D6S1581.</p><p><b>METHODS</b>Genescan was used to genotype D6S1581 of 12 NPC pedigrees, 85 sporadic NPC patients and 181 normal volunteers. Then parametric/nonparametric linkage analysis and association analysis were performed.</p><p><b>RESULTS</b>D6S1581 was linked with NPC, a Lod score of 2.611436 (P=0.00245) was obtained, and a significant difference in allele frequency was observed between familial NPC and control (P<0.005).</p><p><b>CONCLUSION</b>These results suggest that D6S1581 is highly associated with NPC, and there may be one or more NPC associated genes near D6S1581, including FBXO30.</p>

Genetic polymorphism of 8 STR loci on short arm of chromosome 3 / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To get the genotype and allele frequency distributions of 8 short tandem repeat (STR) loci on chromosome 3p (D3S1297, D3S1489, D3S1266, D3S1568, D3S1289, D3S1300, D3S1285 and D3S3681) in Chinese Han population in Hunan area.</p><p><b>METHODS</b>Blood samples were collected from the random Han individuals in Hunan and the whole genomic DNA was extracted. STR loci were amplified by multiplex-PCR technique and genotyped by ABI 377 sequencer.</p><p><b>RESULTS</b>Ninety-one alleles were detected, with frequencies ranging from 0.002 to 0.431, and these alleles constituted 312 genotypes. All the 8 loci met Hardy-Weinberg equilibrium. The statistical analysis of 8 STR loci showed the heterozygosity (H) >or= 0.729, the discrimination power (DP) >or= 0.725, the probabilities of paternity exclusion (PPE) >or= 0.596, and the polymorphic information content (PIC >or= 0.682). The result indicated that there was a significant difference between Han ethnic group and the white and the black.</p><p><b>CONCLUSION</b>These results could serve as valuable data to enrich the Chinese genetic database and play an important role in Chinese population genetic and forensic medical application.</p>

Effect of apelin on human osteoblasts / 中华内分泌代谢杂志

Objective To observe apelin and its receptor (APJ) expressions in human osteoblasts and evaluate the effect of apelin on osteoblasts.Methods The expressions of apelin and APJ in human osteoblasts were tested by RT-PCR and Western blot.After human osteoblasts were treated with apelin,cell proliferation was measured by [~3H] thymidine incorporation and cell counting.Cell function was measured by alkaline phosphatase (ALP) activity,the secreted osteocalcin level and typeⅠcollagen production .The activation of signaling cascades was tested by Western blot.Small-interfering RNA (siRNA) to blockade APJ was applied to observe effects of apelin on cell proliferation and the activation of signaling cascades.Results Both apelin and APJ were expressed in human osteoblasts.Apelin increased the proliferation and did not show the influences on ALP activity, osteocalcin secretion and type I collagen production in human osteoblasts.Apelin induced activation of phosphatidylinositol-3 kinase (PI3K) downstream effector (Akt),but not mitogen-activated protein kinase (MAPK) such as c-jun N-terminal kinase (JNK),p38 and ERK1/2 in human osteoblasts.Suppression of APJ with siRNA or LY294002 (PI3K inhibitor) abolished the apelin-induced cell proliferation and the activation of Akt.Conclusion Human osteoblasts express apelin and APJ.Apelin stimulates the proliferation of human osteoblast via APJ/PI3K/Akt pathway,but has no effect on osteoblast differentiation.