Expression and clinical significance of small fragments of P28 and P43 in papillary thyroid carcinoma after TRa1 gene transcription / 实用肿瘤学杂志

Objective The objective of this study was to investigate the expression of P28 and P43 in papillary thyroid carcinoma(PTC)and its relationship with clinicopathological features after TRa1 gene transcription.Methods Real-time fluorescence quantitative PCR(RT-PCR)and gel electrophoresis were used to determine the target fragments and to isolate the bands of different fragments.The optical density of each band was scanned by UV transmittance analyzer to detect 31 cases of PTC tissue and expression levels of P28 and P43 in para-cancerous tissues.Results The average gray value of P28 in thyroid carcinoma group was 0.77±0.34,which was significantly higher than that in para-cancer group(0.31±0.18).The average gray value of P43(0.85+0.21)in thyroid carcinoma group was significantly higher than that in para-cancer group(0.34±0.15)(P0.05),The expression levels of P43 were not correlated with gender,age,tumor size and lymph node metastasis.(P>0.05)but they were related to clinical pathologic stage(P<0.05).There was no correlation between expression levels of P28 and P43(r=0.266,P=0.071).Conclusion The increased expression of P28 and P43 may have a high degree of malignancy and a certain clinical value in predicting the adverse prognosis of PTC.Both factors are helpful for the prevention and treatment of PTC.

The expression and significance of β-catenin and p28GANK in residual hepatocellular carcinoma cells after transcatheter arterial chemoembolization / 实用肿瘤学杂志

Objective To investigate the expression and significance of β-catenin and p28GANK in residual hepatocellular carcinoma (HCC)cells after transcatheter arterial chemoembolization (TACE).Methods We collected forty-five cases of surgical specimens of hepatocellular carcinoma after TACE ( TACE group ) and thirty cases of surgery without any treatment (pure surgery group).The expression of β-catenin and p28GANK were detected by using immunohistochemical SP method and compared between the two groups .Results The positive expression of β-catenin and p28GANK in TACE group were 77.78%and 75.56%respectively,which were significantly higher than those in pure surgery group (46.67%and 53.33%respectively,P<0.05).In the residual hepatocellular carcinoma cells of TACE group ,the positive expression of β-catenin showed correlation with the positive expression of p28GANK(Φ=0.318,P =0.033).The high expression of β-catenin and p28GANK were closely related to portal vein thrombosis and distant metastasis (P<0.05).Conclusion The ex-pression of β-catenin and p28GANK in the residual hepatocellular carcinoma cells were increased significantly after TACE.The high expression of β-catenin and p28GANK were closely related to portal vein thrombosis and distant metastasis.The high expression of β-catenin and p28GANK may be one of the reasons of hepatocellular carcinoma invasiveness and metastasis .

Sensitivity evaluation of a single-step PCR assay using Ehrlichia canis p28 gene as a target and its application in diagnosis of canine ehrlichiosis / Avaliação da sensibilidade da PCR em uma etapa com base no gene p28 de Ehrlichia canis e sua aplicação no diagnóstico da erliquiose canina

The aim of this study was to optimize a PCR assay that amplifies an 843 pb fragment from the p28 gene of Ehrlichia canis and compare it with two other PCR methods used to amplify portions of the 16S rRNA and dsb genes of Ehrlichia. Blood samples were collected from dogs suspected of having a positive diagnosis for canine ehrlichiosis. Amplification of the p28 gene by PCR produced an 843-bp fragment and this assay could detect DNA from one gene copy among 1 billion cells. All positive samples detected by the p28-based PCR were also positive by the 16S rRNA nested-PCR and also by the dsb-based PCR. Among the p28-based PCR negative samples, 55.3 percent were co-negatives, but 27.6 percent were positive in 16S rRNA and dsb based PCR assays. The p28-based PCR seems to be a useful test for the molecular detection of E. canis, however improvements in this PCR sensitivity are desired, so that it can become an important alternative in the diagnosis of canine ehrlichiosis.

O objetivo deste estudo foi aperfeiçoar um ensaio de PCR que amplificasse um fragmento de 843 pares de bases do gene p28 da Ehrlichia canis e compará-lo com outros dois métodos de PCR utilizados para amplificar partes do gene 16S rRNA e dsb do gênero Ehrlichia. Amostras sanguíneas foram colhidas de cães com diagnóstico clínico de erliquiose. A amplificação do gene p28 pela PCR produziu um fragmento de 843pb e esse ensaio permitiu a detecção do DNA de um parasita dentre 1 bilhão de células. Todas as amostras positivas detectadas pela PCR baseada no gene p28 foram também positivas pela nested PCR para detecção do gene 16S rRNA e também pela PCR dsb. Dentre as amostras negativas para a PCR p28, 55,3 por cento foram co-negativas, mas 27,6 por cento foram positivas pela PCR baseada nos genes 16S rRNA e dsb. A PCR p28 parece ser um teste útil para detecção molecular de E. canis, entretanto otimizações na sensibilidade nesta PCR são necessárias, para que esta técnica se torne uma importante alternativa no diagnóstico da erliquiose canina.

Construction of oncogene p28GANK mutants and their regulating effects on p53 / 第二军医大学学报

Objective: To construct the mutants of oncogene p28GANK with different ankyrin repeats deleted, so as to further study the potential role of p28GANK in hepatocellular carcinoma (HCC). Methods: QuikChang Site-Directed Mutagenesis Kit was used to construct five p28GANK mutants with different ankyrin repeats deleted and their influences on p53 were also investigated. Results: Five deletion mutants of p28GANK were as follows: 39-71aa, 72-104aa, 105-137aa, 138-170aa, and 171-203aa. Agarose gel electrophoresis and Western blotting analysis confirmed the correct construction of these mutants. Wild type p28GANK promoted degradation of p53, but the 5 mutants had no noticeable effects on expression of p53. Conclusion: We have successfully constructed five p28GANK ankyrin repeat-deleted mutants. It is confirmed that the regulation of p53 needs the complete ankyrin repeats of p28GANK, which paves a way for further research on ankyrin and the role of p28GANK in HCC.