Effects of miR-181 b on proliferation and apoptosis of thyroid papillary carcinoma cells by down-regulating CYLD protein / 局解手术学杂志

Objective To investigate the effect of miR-181b on the proliferation and apoptosis of thyroid papillary carcinoma cells by down-regulating CYLD protein and its mechanism. Methods qPCR was used to detect the expression and difference of miR-181b in papilla-ry thyroid carcinoma and normal thyroid tissue. Western blotting was used to detect the regulation between miR-181b and CYLD protein. Clone formation assay was used to detect the proliferation of thyroid cancer cells after inhibition of miR-181b. Flow cytometry was used to detect the apoptosis of thyroid carcinoma cells after inhibition of miR-181b. Results Compared with normal thyroid tissue,the expression of miR-181b in papillary thyroid carcinoma was up-regulated,the difference was statistically significant(P<0. 05). The expression level of miR-181b in FTC-133 cell line was relatively high. Western blotting confirmed that miR-181b could directly regulate the expression of CYLD protein. Inhi-bition of miR-181b expression can inhibit the proliferation of thyroid cancer cells,and to some extent promote its apoptosis behavior. Conclu-sion miR-181b can regulate the expression of CLYD and affect the proliferation and apoptosis of thyroid cancer cells.

Asiatic acid mitigates hyperglycemia and reduces islet fibrosis in Goto-Kakizaki rat, a spontaneous type 2 diabetic animal model / 中国天然药物

The Goto-Kakizaki (GK) rat is a spontaneous type 2 diabetic animal model, which is characterized by a progressive loss of beta islet cells with fibrosis. In the present study, the hypoglycemic effect of asiatic acid (AA) in GK rats was examined. GK rats receiving AA at a daily dose of 25 mg·kg(-1) for four weeks showed a significant reduction in blood glucose levels. Age-matched normal Wistar rats were given 0.5% sodium carboxymethyl cellulose (CMC-Na) solution for the same periods and used as control. Compared to the normal Wistar rats, GK rats treated with AA showed improvement in insulin resistance partially through decreasing glucose level (P < 0.01) and insulin level (P < 0.05). Furthermore, the results of immunohistochemistry indicate that AA treatment reduced islet fibrosis in GK rats. Fibronectin, a key protein related to islet fibrosis, was over-expressed in GK rats, which was reversed significantly by AA treatment (P < 0.05). These findings suggest that AA has a beneficial effect on lowering blood glucose levels in GK rats and improves fibrosis of islets in diabetes, which may play a role in the prevention of islets dysfunction.

Isolation, culture and identification of human odontoclasts / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To isolate, culture and identify odontoclasts in vitro and to establish a method of culturing human odontoclasts.</p><p><b>METHODS</b>Healthy and retentive deciduous teeth were extracted, and then placed in α-minimum essential medium containing 0.1% collagenase and 0.2% dispase for 1 h.Odontoclasts were obtained and incubated from the absorbing root surfaces of deciduous teeth.Isolated cells were viewed by inverted phase contrast microscope firstly. Then, the isolated odontoclasts were morphologically observed by hematoxylin and eosin staining (HE) and tartrate-resistant acid phosphatase (TRAP) staining. The prepared teeth slices were cocultured with the isolated odontoclasts and scanning electronic microscope(SEM) was used to demonstrate the presence of resorption lacunae.</p><p><b>RESULTS</b>The isolated odontoclasts appeared as multinucleated giant cell with many vacuolus in cytoplasm. TRAP staining demonstrated that the cytoplasm of the odontoclasts was full of claret-red positive particles.Resorption lacunae on teeth slices which cocultured with odontoclasts were seen under SEM.</p><p><b>CONCLUSIONS</b>Enzyme digestion is an effective method to isolate odontoclasts from absorbing root surface of deciduous teeth.</p>

Development of Chinese version of children's fear survey schedule-dental subscale / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To establish the Chinese version of the modified children's fear survey schedule-dental subscale (CFSS-DS).</p><p><b>METHODS</b>The original English version of CFSS-DS with facial image scale (FIS) was translated into Chinese, pre-tested and cross-culturally adapted. Subsequently the Chinese version schedule was randomly investigated among 367 children aged 5 - 12 years and their parents. Reliability and validity of the translated scale was evaluated later.</p><p><b>RESULTS</b>Totally 367 children were investigated and 311 valid questionnaire was received. Cronbach's alpha of the translated scale was 0.85 and test-rest reliability was 0.73. The 15 items were divided into four domains. There was a certain logical relationship between the items among the same domains. There was highly significant association between the self-report of the Modified CFSS-DS and Frankl Behavioral Scale (r(s) = -0.403, P < 0.01).</p><p><b>CONCLUSIONS</b>The Chinese version of modified CFSS-DS has been established successfully with good psychometric properties which provide the theoretical evidence for further application in Chinese population.</p>

Effects of guanine-quadruplexes formation induced by adriamycin on telomeric extension reaction mediated by telomerase of Tca8113 cells / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To study the effects of adramycin to disturb telomeric extention reaction mediated by telomerase of Tca8113 cells by inducing oligonucleotides that contain telomeric repeats to form guanine-quadruplex (G4) structures.</p><p><b>METHODS</b>In the presence of adriamycin, d(TTAGGG)4, d(TTAGAG)4, d(TTAGGG)5 and d(TTAGGGT) were analyzed by electrophoretic mobility shift assay. The mobility of d(TTAGGG)3, d(TTAGGG)4 and d(TrAGGG)5 in native polyacrylamide electrophoresis were observed. Methylation protection experiments were performed to investigate the effects of adriamycin on methylation of guanine in d(TTAGGG)4 and d(TTAGAG)4. The traditional telomeric repeats amplification protocol (TRAP) and modified TRAP-G4 assays were, respectively, used to analyze the different characteristcs of adriamycin's inhibiting telomeric extension mediated by telomerase of Tca8113 cells.</p><p><b>RESULTS</b>At 5.00 microg/mL of adriamycin, conversion of some of linear d(TrAGGG)4 and d(TrAGGG)5to the new, high-mobility bands formed by complex with special second structures were found in the mobility shift assay. Adriamycin at 1.25 microg/mL protected the G in d(TIAGGG)4 from methylating. Adriamycin at 2.50 microg/mL or 1.25 microg/mL partially inhibited the telomeric extension lengthened by telomerase of Tca8113 cells in TRAP assay, but completely did so in TRAP-G4 assay.</p><p><b>CONCLUSION</b>Adriamycin is able to disturb telomeric extention mediated by telomerase of Tca8113 cells by inducing oligonucleotides that contain telomeric repeats to form intra-molecular G4 structures.</p>

Radiation-inducible promoters-mediated cdglytk gene in the treatment of buccal carcinoma in golden hamster / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To observe the therapeutic effect of CDglyTK gene mediated by radiation-inducible promoters in the treatment of buccal carcinoma in Golden Hamster.</p><p><b>METHODS</b>Animal models of buccal carcinoma in golden hamster were established by painting 0.5% dimethyl-benzanthracene. The plasmids pcDNA (+) 3.1/E-CDglyTK were transfected into tumors by lipofectamine. 24 h later, the tumors were exposed to 3 Gy irradiation. Animals were monitored at regular intervals for volume of tumors. CDglyTK mRNA was assayed by RT-PCR. Apoptosis and proliferating cell nuclear antigen were detected respectively by in situ end-labeling and immunohistochemical methods.</p><p><b>RESULTS</b>Compared with control groups, the tumor was suppressed obviously by CDglyTK gene therapy combined with 3 Gy induction radiation. The expression of CDglyTK gene could be detected by RT-PCR in the transfected tumor, and up-regulation of CDglyTK expression was found in tumor exposed to radiation (P < 0.05). There was significant difference in apoptosis index or proliferation index between tumor without irradiation and tumor with irradiation (P < 0.05).</p><p><b>CONCLUSIONS</b>The radiation-inducible promoter can be served as a molecular switch to regulate the expression of CDglyTK gene in buccal carcinoma in golden hamster, and low dose induction radiation can significantly improve the therapeutic effects.</p>

Synthetic radiation-inducible promoter mediated CDglyTK gene in treatment of Tca8113 cells / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To observe the therapeutic effect of CDglyTK gene mediated by synthetic radiation-inducible promoter in the treatment of Tca8113 cells.</p><p><b>METHODS</b>CDglyTK gene in pCEA-CDglyTK was subcloned into pcDNA3.1 (+) to construct plasmid pcDNA3.1 (+)-CDglyTK, and then the synthetic radiation-inducible promoter in pMD18 -T -E was inserted into pcDNA3.1 (+) -CDglyTK to construct plasmid pcDNA3.1 (+ )/E -CDglyTK. The recombinant plasmid was transfected into Tca8113 cells by lipofectamine, and then exposed to 3 Gy irradiation. Cytotoxicity was evaluated by MTT. The expression of CDglyTK gene was detected by RT-PCR. The apoptosis and proliferation were examined by flow cytomtery.</p><p><b>RESULTS</b>The plasmid pcDNA3.1 (+)/E-CDglyTK was constructed successfully. The comparative survival rate of Tca8113 cells was markedly decreased by induction irradiation. Up-regulation of CDglyTK expression was found in Tca8113 cells exposed to irradiation. The apoptosis index (AI) of Tca8113 cells exposed to irradiation was higher than that of Tca8113 cells without irradiation, the other way round, the proliferation index (PI) of Tca8113 cells exposed to irradiation was lower than that of Tca8113 cells without irradiation.</p><p><b>CONCLUSION</b>The synthetic radiation-inducible promoter can be served as a molecular switch to improve the expression of CDglyTK gene in Tca8113 cells, and low dose induction radiation can significantly improve the therapeutic efficiency.</p>