1.Demethylation stduy of decitabine in acute leukemia patients with Apaf-1 gene methylated
Jun GUO ; Enqin YANG ; Hui YANG
Journal of Leukemia & Lymphoma 2013;22(9):538-541
Objective To investigate the effectiveness of decitabine demethylation in treatment of acute leukemia.Methods Methylation specific PCR (MSP) was used to detected the methylation status of Apaf-1 gene promoter.10 cases entering the group.MSP was used to detected the 10 cases methylation status of Apaf-1 promoter between pre-and post-treatment of dicitabine.RT-PCR method used was to detect the differential expression levels of Apaf-1 mRNA in acute leukemia bone marrow mononuclear cell between preand post-treatment of decitabine.Results In post-treatment of decitabine,6 cases Apaf-1 gene promoter was demethylated.The loss expression of Apaf-1 mRNA re-expressed in 4 cases.6 cases Apaf-1 mRNA still express deletion.6 cases patients have Apaf-1 mRNA exprssion deletion,However,4 cases Apaf-1 gene was demethylated,2 cases methylated in post-treatment,maybe related to allele deletion or allelic varriants.Conclusion Post treatment of decitabine.Apaf-1 gene promotor was demethylated and repress the expression of Apaf-1 mRNA,play a key role in apoptosis maybe a new method for treatment of acute leukemia.
2.Determination of Ephedrine Hydrochloride in Zhike Pingchuan Tangjiang by HPLC
Xin-Hui GUO ; Fen WANG ; Jun-An CAI ;
Chinese Journal of Information on Traditional Chinese Medicine 2006;0(11):-
Objective To establish the method for detemining the content of ephedrine hydrochloride in Zhike Pingchuan Tangjiang by HPLC. Methods Diamonsil ODS1 C_(18) Column was used with acetonitrile -0.1% phosphoric solution (with 0.1% triehylamine) (3 : 97) as the mobile phase, the detection wavelength as 205 nm, and flow rate was 1.0 mL/min. Results The calibration curve was linear at the range of 0.12~ 0.96 ?g for ephedrine hydrochloride and linear equation was Y= 109759X+3792.8, r=0.9998. The average recovery was 98.4% and RSD was 0.87% (n =5). Conclusion This method was simple, accurate and proper, with good reproducibility. It can be used for quantitative analysis of ephedrine hydrochloride in Zhike Pingchuan Tangjiang.
3.CD4+CD25+FoxP3+Treg in the immune mechanism of severe mycoplasma pneumoniae pneumonia in children
Yun GUO ; Jun QIAN ; Ling LI ; Yu HUI ; Hanmin JIANG
International Journal of Pediatrics 2016;43(8):647-650
Objective To investigate the role of CD4+CD25+FoxP3+ in severe Mycoplasma pneumonia among children.Methods One hundred and forty children with M.pneumoniae pneumonia (65 severe and 75 non-severe) who were hospitalized were enrolled along with forty other children as controls.X-ray was assessed.The proportions of peripheral blood CD4+CD25+FoxP3+cells were determined by flow cytometry.Results Both severe and non-severe children had decreased CD4+CD25+FoxP3+cells as compared with control subjects in acute phase (0.87 ± 0.66% vs.3.88 ± 2.00%,P < 0.01 and 1.17 ± 0.70% vs.3.88 ±2.00%,P <0.01,respectively).The levels of CD4+CD25+FoxP3+cells in severe children were lower than those in non-severe children in acute phase and recovery phase (0.87 ±0.66% vs.1.17 ±0.70%,P <0.05 and 1.66 ±0.85% vs.3.61 ± 1.45%,P<0.01,respectively).Both severe children and non-severe children expressed higher CD4+CD25+FoxP3+cells in recovery phase than in acute phase (1.66 ± 0.85 % vs.0.87 ± 0.66%,P <0.01 and 3.61 ± 1.45% vs.1.17 ±0.70%,P <0.01,respectively).Conclusion The expression of CD4+CD25+FoxP3+Tregs may play a role in the onset of severity of mycoplasma pneumonia and the low express of CD4+CD25+FoxP3+Tregs in children infected with M.pneumonia may increase the susceptibility to severe mycoplasma pneumonia.
9.Implication of expression of Nanog in prostate cancer cells and their stem cells.
Chen, GONG ; Hui, LIAO ; Fengjin GUO ; Liang, QIN ; Jun, QI
Journal of Huazhong University of Science and Technology (Medical Sciences) 2012;32(2):242-6
Recent studies suggested that the prostate cancer may arise from prostate cancer stem cells that share some same characteristics with normal stem cells. The purpose of this study was to detect the differences of Nanog expression between PC3 prostate cancer cell line and its tumor stem cells, and the relationship was preliminarily examined between Nanog and prostate cancer and its tumor stem cells. By using magnetic active cell sorting (MACS), we isolated a population of CD44(+)/CD133(+) prostate cancer cells that display stem cell characteristics from PC3 cell line. Immunohistochemistry revealed positive expressions of CD44, CD133 and α(2)β(1)-integin in the isolated cells. CCK-8 analysis showed that isolated cells had a strong proliferative ability. The formation of the cell spheres in serum-free medium and holoclones in serum-supplied medium showed that the cells were capable of self-renewing, indicating that the isolated cells were a population of cancer stem-like cells derived from PC3 cell line. Western blotting exhibited that the isolated cells had higher experession of Nanog, an embryonic stem marker, as compared with PC3 cells. Our study showed that Nanog might be helpful in sustaining the self-renewal and the undifferentiation of prostate cancer stem cells, and may serve as a marker for prostate cancer stem cells for isolation and identification.
10.Effect of Licofelone on Expression of Fractalkine Induced by Interleukin-18 in Mesangial Cells
hui, CHEN ; yuan-jun, WU ; guo-ping, ZHOU
Journal of Applied Clinical Pediatrics 2006;0(23):-
Objective To study the effect of Licofelone,a novel non-steroid anti-inflammatory drug,on the expression of Fractalkine induced by interleukin-18(IL-18) in mesangial cells.Methods Rat mesangial cells were cultured and divided into IL-18 stimulated group,Licofelone-treated group and normal control group.The cells in IL-18 stimulated group were stimulated by 10 ?g/L IL-18 for 24 h.In Licofelone-treated group,ahead of exposure of IL-18 for 24 h,cells were treated with Licofelone in the doses of 10,50 and 100 ?mol/L for 30 min.Additionally,the mesangial cells without treatment of IL-18 and Licofelone were used as normal control group.Reverse transcription-polymerase chain reaction(RT-PCR) was used to measure the level of Fractalkine mRNA.The expressions of Fractalkine protein in every group were detected with enzyme linked immunosorbent assay (ELISA).Results In normal control group,the expression level of Fractalkine mRNA was 179.0?21.0.After exposure of IL-18 for 24 h,the level of Fractalkine mRNA was 1 220.1?185.7,which was higher than that in normal control group (t=9.646 P