1.Comparison of telomerase, p53 and Ki-67 expression on colorectal serrated adenoma, traditional adenoma and colorectal cancer
Youming WU ; Yanqing DING ; Na LI ; Jinsheng WANG ; Zhouqing GUO ; Lei LENG ; Dong HUANG
China Oncology 2013;(7):519-523
Background and purpose:Colorectal serrated adenoma (SA) was ofifcially named in 2000 by the WHO as a separate disease, with unique properties compared with traditional adenoma (TA), and its relationship with colorectal cancer (CRC) is very concerned. This study was to analyze and compare the telomerase, p53 and Ki-67 immunohistochemical expression on the tissues of SA, TA and CRC. Methods:Immunohistochemistry was adopted to analyze the expression of telomerase, p53, and Ki-67 in 37 cases of SA, 36 cases of TA and 34 cases of CRC. Results:The p53-positive percentage of SA was signiifcantly lower than that of TA (P<0.01), and the p53-positive percentage of TA was signiifcantly lower than that of CRC (P<0.01). No signiifcant difference of Ki-67 expression was found between SA and TA, and the Ki-67-positive percentage of SA and TA was lower than that of CRC (P<0.01). The telomerase-positive percentage of TA was signiifcantly lower than that of SA (P<0.01), and the telomerase-positive percentage of SA was signiifcantly lower than that of CRC (P<0.05). Conclusion:Telomerase, P53, and Ki-67 immunohisto chemical analysis indicated that SA is a kind of proliferative adenoma, and telomerase activation may play a role in the cancer process.
2.Pyrosequencing of pre-C/BCP mutation in hepatitis B
Lan YAO ; Jiawu SONG ; Zhi XIN ; Xiaofeng LI ; Zhouqing WU ; Jianxi TANG ; Bo GUO ; Bo WU ; Renxu LAI
Chinese Journal of Laboratory Medicine 2008;31(8):860-863
Objective To develop a clinically useful assay for detecting the mutations of HBV pre-C/BCP based on the pyrosequencing and accuracy, reproducibility and reliability of this assay was evaluated. Methods The pyrosequencing primers for HBV pre-C/BCP mutation were designed through the cluster analysis among one hundred HBV gene sequences. After the amplification of the fragment of pre-C/BCP with the template of pre-C/BCP mutation plasmids, the pyrosequencing method for pre-C/BCP detection was initially set up with this standard sample. The accuracy, reliability and reproducibility of the pre-C/BCP pyrosequencing were confirmed through the pre-C/BCP plasmids as a standard sample when compared with Sanger/genechip sequencing method pre-C/BCP pyrosequencing assay was applied for detecting pre-C/BCP mutation types of 60 chnical serum samples in HBV patients. Results The pre-C/BCP mutation detection assay based on pyrosequencing has been established in our study. The coincidence rate between pyrosequencing and Sanger squencing was 100%. The coincidence rate between the result of pyrosequencing and of genechip method was 91.7%. The reproducibility of this assay was 97. 8%. It indicates the pre-C/BCP pyrosequencing is a high-accurate method with, good-reproducibility and high-reliability. And multi-site detection can be achieved by pyrosequencing one time. A rare mutation T1758C was also detected. Conclusion Pyrosequencing for pre-C/BCP mutations assay is high-throughout method for simultaneous detection of multi-site mutation.