Association of EGFR gene G719S and T790M mutations with cervical cancer / 中华医学遗传学杂志

OBJECTIVE@#To establish a rapid and accurate "on/off" switch technique consisted of 3'-phosphorothioate-modified allele-specific primers and exo+ polymerase to screen the G719S and T790M mutations of epidermal growth factor receptor (EGFR) gene. The switch was used to identify cervical cancer patients who are sensitive to tyrosine kinase inhibitor (TKI).@*METHODS@#Allele-specific primers targeting recombinant wild-type and mutation-type templates were designed with 3' terminal phosphorothioate modification. Two-directional primer extension was carried out using Pfu polymerase. The G719S and T790M mutations were detected by the technique among cervical cancer tissues. The results were verified by Sanger sequencing.@*RESULTS@#No mutation was detected among the 80 cervical cancer cases, and the results were consistent with that of Sanger sequencing. No significant difference was found between the frequencies of the G719S and T790M mutations between the patient and the control groups (P> 0.05).@*CONCLUSION@#A sensitive "on/off" switch technique for detecting the two EGFR mutations was established. The G719S and T790M mutations are not associated with cervical cancer.

Construction of fused recombinant vector of EGFR gene containing G719S and T790M by overlap PCR / 临床检验杂志

Objective To construct the recombinant pMD19-exon18-exon20 plasmid containing locus G719S and T790M of EGFR gene associated with cervical cancer,which may provide a template for preparing the mutant recombinant vector of EGFR gene.Methods Using the healthy human genome DNA as templates,the segments of exon 18 and exon 20 of EGFR gene were amplified by two pairs of specific primers which were designed based on the sequences of overlapping and complementary area.The amplified segments were linked by overlap PCR.The products of linked exon18-exon20 were further inserted into the vector pMD19-T.The constructed recombi nant pMD19-exon18-exon20 plasmid was finally transformed into competent cells E.coli DH5α and then identified by PCR with bacterial solution and genome sequencing.Results The amplified fragments of exon18 and exon20 were clearly appeared at 778 bp and 596 bp and the fused product of exon18-exon20 was showed at 1 374 bp on agarose gel electrophoresis.The recombinant plasmid of fusion EGFR gene was consistent with the expected results via bacterial PCR assay and DNA sequencing.Conclusion We successfully fused the segments of exon18 with exon20 and constructed the recombinant expression vector of EGFR gene by using overlap PCR method.

Clinical and genetic evaluation of a family with isolated oligodontia / 实用医学杂志

Objectives To investigate the mutational characteristics of MSX1 and PAX9 genes in a family affected by non-syndromic oligodonti so as to study the pathogenesis of oligodontia from a molecular prospective. Methods A family with oligodontia, but of different descent and unrelated healthy controls were enrolled in our study. Genomic DNA was isolated from the blood samples. Mutation analyses were performed by amplifying MSX1 and PAX9 exons and sequencing the products. Results DNA sequencing revealed a novel missense mutation c.348C>T in a highly conserved homeobox sequence of MSX1 and a known polymorphisms c. 469+35- c.469+45del in exon 1 and in intron in the two patients and in two unrelated healthy controls. But we did not detect any mutation in PAX9. Conclusion Our finding suggests the samesense mutation (c.348C>T) and the polymorphisms (c.469+35- c.469+45del) may be responsible for oligodontia phenotype in this Chinese family.

The effects of periodontal therapy on serum lipid level in patients with periodontitis and hyperlipidemia / 实用口腔医学杂志

Objective:To evaluate the effects of periodontal treatment with scaling and root planning supplemented with chlorhexidine on plasma lipids in patients with chronic periodontitis and hyperlipidemia(CPH).Methods:6 1 patients with CPH were randomly as-signed into control-treatment group(n=30)and intensive-treatment group(n=31).Patients in the control-treatment group underwent a standard cycle of supragingival mechanical scaling and polishing.Patients in the intensive-treatment group underwent the adjunctive full-mouth intensive removal of subgingival dental plaque biofilms with the use of scaling and root planning.Periodontal parameters,to-tal cholesterol(TC),triglycerides(TRG),high-density lipoprotein(HDL)and low-density lipoprotein(LDL)were measured before treatment and 2 and 6 months after treatment.Results:2 and 6 months after treatment,TRG was significantly lower in the intensive-treatment group than that in the control-treatment group(P<0.05),the levels of HDL were significantly higher(P<0.05);however, the levels of total cholesterol and LDL,in the intensive-treatment group were not significantly different from those in the control-treat-ment group 2 and 6 months after therapy.Conclusion:Intensive periodontal treatment may improve serum lipid levels in patients with hyperlipidemia and chronic periodontitis.