Biomedical subjects

Bu-ling Wu

Publications and source records attributed to Bu-ling Wu.

8 recordsLinked to original sources

PubMed2006

[Generation of transgenic mouse of dentin sialoprotein and transgene expression analysis].

PURPOSE: To generate the transgenic mouse model of DSP and perform transgene expression analysis by RT-PCR. METHODS: Plasmid pcDNA3.1-CX was constructed by substituting promoter cbeta-actin for CMV promoter of pcDNA3.1, and the ultimate transgenic vector, pcDNA3.1-CX-dsp, was constructed by cloning DSP coding sequence into pcDNA3.1-CX. The pcDNA3.1-CX-dsp plasmid was linearized and microinjected into the male pronucleus of the zygotes. The tail DNA of pups was tested by PCR and Southern blot. A member of F1 generation of one positive mouse was used to perform transgene expression analysis by RT-PCR. RESULTS: 717 embryos were implanted to 29 recipient pseudopregnant mice, 4 of the 67 pups carrying the transgene. Expression of DSP was detected in a member of F1 generation of one positive mouse by RT-PCR. CONCLUSION: Founders of the DSP transgenic mouse were obtained successfully, and the expression of DSP was primarily confirmed.

Animals↗

PubMed2005

[Generation of dspp-LacZ transgenic mouse founders].

PURPOSE: To establish a transgenic mouse founders in which the expression of LacZ was directed by a dentin sialophosphoprotein-specific promoter. METHODS: The DSPP-specific promoter was obtained by PCR and confirmed by sequencing, and the transgenic plasmid, pTN-DPM-LacZ, was constructed by subcloning the DSPP-specific promoter and LacZ-encoding sequence into one vector. The linearized transgenic plasmid was microinjected into the male pronucleus of the zygotes, and the microinjected zygotes were implanted to recipient pseudopregnant mice. The tail DNA of 4 week-pups was tested by PCR. RESULTS: 503 embryos were implanted to 20 recipient pseudopregnant mice, 12 of the 89 pups carrying the transgene. The establishment of the dspp-LacZ transgenic mouse line is under progress. CONCLUSION: 12 founders of the dspp-LacZ transgenic mice, in which the expression profile of LacZ should be the same as that of DSPP, were obtained by microinjection successfully, and the mouse line which is being established could be used as a good tool to investigate the exact expression profile of DSPP in the future.

Animals↗

PubMed2005

[Immunization with the fusion protein of GBD of Streptococcus mutans glucan binding protein-A against dental caries in SD rats].

PURPOSE: To observe the effect of immunization with the fusion protein of GBD of Streptococcus mutans glucan binding protein-A against dental caries. METHODS: Purified fusion protein of GBD of Streptococcus mutans glucan binding protein-A was used to immune SD rats by subcutaneous injection route. The rats were fed with Keyes Diet 2000 and infected by S.mutans. The caries level was determined and the result was analyzed by t test. RESULTS: The caries score of SD rats decreased in the group of immunized with GBD fusion protein,P<0.01. CONCLUSION: Immunization with GBD fusion protein resulted in significantly reduced dental caries after infection with S.mutans Ingbritt. Supported by PLA Tenth Five-Year Key Project (01Z089).

Animals↗

PubMed2005

[Cloning and transcriptional regulation analysis of amelogenin promoter in different cells].

PURPOSE: To clone different promoter region of amelogenin gene and analyze their transcriptional activity. METHODS: The upstream regulation sequences of amelogenin gene were retrieved and analyzed. Amplified by PCR from the genomic DNA of mouse C57BL/6J and digested with restriction endonucleases enzyme, different transcriptional regulation sequences of 5'flanking of amelogenin gene including the basal promoter were cloned and ligated with luciferase gene in PGL3-Basic vector. These report vectors were transiently transfected into CHO, Hela and UMR-106 cells and luciferase assay was performed to analyse the transcription activation of these promoters. RESULTS: 6 promoters different in length were cloned. The activity of luciferase was very strong in Hela cells. On the contrary, the CHO and UMR-106 cells showed weak fluorescence. Luciferase activity fluctuated with the different promoter lengths in Hela cell as well as in CHO and UMR-106 cells. 975 bp and 532 bp of amelogenin 5'flanking DNA had a strong transcriptional activation, but 285 bp of amelogenin 5'flanking DNA had a weaker transcriptional activation. CONCLUSION: The sequence of amelogenin promoter can be activated in Hela cell. Hela cell can be used as a good model to study the transcriptional regulation of amelogenin promoter. According to the different activities of different lengths, it is suggested that there were some potential siliencer located between -975 and -532, and some potential activator in the region between -532 and -285.

Amelogenin↗

PubMed2005

[Reconstruction of dentin-pulp complex structure by tissue engineering technology].

OBJECTIVE: To investigate the possibility of reconstruction of dentin-pulp complex by tissue engineering technology. METHODS: Rat dental pulp stem cells were seeded into HA-TCP scaffold and incubated for 20 hours in vitro. Then the cell-scaffold complex was implanted subcutaneously into the dorsal side of nude mice. 8 weeks postimplantation, the samples were extracted for histological and immunohistochemical examinations. RESULTS: Three strata of tissue were observed in the hole of HA-TCP scaffold. They were dentin-like tissue, predentin-like tissue and pulp-like tissue respectively from the inner surface of the pore to the center. Dentin tubules were obvious in predentin-like and dentin-like tissue lining from the pulp-like tissue through predentin-like tissue and dentin-like tissue. Cells localized along the edge of pulp-like tissue were dense and polarized, resembling odontoblasts. Immunohistochemical study demonstrated DSP and DMP1 expression in these odontoblast-like cells and in the area of predentin-like tissue. CONCLUSIONS: Tissue-engineered rat dentin-pulp complex was reconstructed by seeding HA-TCP scaffold with rat dental pulp stem cells.

Animals↗

PubMed2004

[Expression of GBD gene of Streptococcus mutans glucan binding protein A in mammalian cells].

OBJECTIVE: To evaluate the expression of recombinant plasmid pcDNA3.1/GBD of glucan binding protein of Streptococcus mutans in mammalian cells COS-7. METHODS: Eukaryotic plasmid carrying encoding gene of GBD of Streptococcus mutans gbpA was constructed and the plasmid was introduced into COS-7 cells by Lipofectamine reagent. The transient expressed protein in COS-7 cells was detected by immunochemistry technique. RESULTS: The positive expression was detected in plasma of the cells which were transfected with recombinant plasmid pcDNA3.1/GBD. The cells which were transfected with pcDNA3.1 were negative. CONCLUSION: GBD can translate and express in COS-7 cells after transfected with recombinant plasmid pcDNA3.1/GBD. The expressed protein locates in the plasma and the protein is able to combine with anti-GbpA antibody. The expressed protein has the antigenicity and is a candidate gene vaccine.

Animals↗

PubMed2004

[Characterization of Notch gene involved in genetic regulatory networks in dental pulp stem cells].

OBJECTIVE: To investigate the characterization of Notch gene involved in genetic regulatory networks in dental pulp stem cells. METHODS: The pulp tissue was separated from mouse teeth and digested by collagenase type I. Single-cell suspensions of dental pulp were seeded into 6-well plates with alpha modification of Eagle's medium supplemented with ES cell qualified Fetal Bovine Serum. Colony-forming efficiency was assessed in 14ds culture. Transcripts for Notch were detected by reverse transcription-PCR by using total RNA isolated from cells. RESULTS: There were clonogenic cells in dental pulp cell and the incidence of colony-forming cells derived from mouse dental pulp cells was 1.6-2.5 colonies/10(4) plate. Mouse-specific Notch mRNA expressed in colony-forming cells. CONCLUSION: Notch mRNA expressing in colony-forming cells provided a more detailed understanding of mouse dental pulp stem cell biology.

Animals↗

PubMed2003

[Cbfa1 induces the expression of the mineral-related proteins in human dental papilla cells].

OBJECTIVE: To explicit whether the expression of the mineral-related proteins is regulated by cbfa1 in human dental papilla cells. METHODS: Human 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. RESULTS: We 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. CONCLUSIONS: In 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↗