Search Results

1.Analysis of protein expression in retinoic acid-induced HL60 cells by modified two--dimensional electrophoresis

Journal of Chongqing Medical University 2007;0(12):-

Objective:To establish the granulocyte-differentiation model of the HL60 cells which are treated with All-trans retinoic acid(ATRA),and to use the modified two-dimensional electrophoresis conditions to analyze the differences of protein expression between treated and untreated HL60 cells.Methods:HL60 cells were induced through treatment with All-trans retinoic acid(ATRA).For selection of the appropriate drug concentration and induction time,MTT and flow cytometry are used to detect the HL60cell proliferation and the expression of differentiation antigens CD11b respectively.Cellular chemical staining was used for the verification of the differentiation of the treated HL60 cells.The protein of HL60 cell lines could be separated by modified two-dimensional electrophoresis(2-DE).PDQuest software was used to analyze the different protein expression between treated and untreated HL60 cells.The protein was analyzed by matrix-assisted laser desorption -time of flight mass spectrometry(MALDI-TOF).Results:ATRA could inhibit HL60 cell proliferation,and with the increase in drug concentration,the effect of inhibiting was more significant.Treated with 2? M ATRA for five days,there were more than 90% of HL60 cells expressing antigenCD11b.Cellular chemical staining also showed that ATRA could induce HL60 cells to granulocyte cells.By the analysis of modified 2-DE and PDQuest software,25 protein spots was detected in untreated cells,while 15 protein spots was promoted Some of them were oncogene protein and suppressor gene protein,while some of others are involved in apoptosis.Conclusion:ATRA could induce HL60 cells to granulocyte cells in selected drug concentration and induction time.Using the modified two-dimensional electrophoresis conditions,different protein expression can be found from the traditional two-dimensional electrophoresis.

Objective To determine and evaluate the method of localization of anal verge by mutislice spiral CT. To provide an imaging reference for operative guidance of low-rectal cancer. Methods Forty eight consecutive adult patients suspected of abnormalities other than rectal disease were evaluated with abdominal and pelvic CT scans since August, 2009. They were divided into two groups based on sex and age. There were 23 men and 25 women. The ages of young group were 28 to 50 years and the average age was 41 years. The ages of elderly group were 52 to 81 years and the average age was 64 years. A small cotton ball dipped with contrast media was put at the anal verge as a marker and CT scans were performed with 64-slice spiral CT scanner. The distances between the cotton balls and the lower margin of the pubis combination (La), the lower margin of the 5th sacral vertebra (Lb), the inferior aperture of minor pelvis(Lc) and the lower margin of the basement of external anal sphincter (Ld) were measured on the midsagittal images obtained by MPR. The averages, the standard deviations(s), the 95% and 80% confidence intervals of La, Lb, Lc and Ld were calculated. We took the intervals of ± 1.96 s or ± 1.28 s ＜ 5 mm as the standard of meeting the clinical requirement. The distances meeting the clinical requirement between different sex groups or age groups were compared with t test. Results The Ld was (9.8 ± 1.2) mm, ofwhich intervals of ± 1.96 s and ± 1.28 s were 4. 8 mm and 3.0 mm respectively, meeting the clinical need.This distance was also measured conveniently. There were no significant differences ( P ＞ 0.05 ) between two different sex groups [ male group, ( 10.0 ± 1.2) mm], female group, (9.6 ± 1.2) mm and between two age groups [ young group, (9.6 ± 1.2) mm, elderly group, ( 9.9 ± 1.3 ) mm ]. Conclusions The lower margin of the basement of external anal sphincter was a useful anatomic lanmark for localizing the anal verge,and could be definitely identified on the middle sagittal pelvic CT image. The distance between the structure and anal verge is constant enough and can be used in measuring distance from low rectal lesion to the anal verge.