Tyrosine kinase inhibitor (TKI) may significantly improve the treatment outcome in chronic myeloid leukemia (CML).It is the most frequent question about whether the patients with durable complete molecular response (CMR) can safely discontinue TKI treatment without relapse.This has focused attention on the strategies to eradicate residual CML cells,especially the CML stem cells,which should result in long term leukemia-free survival and permanent cure.Here,the progress on discontinuation of TKI therapy and alternative approaches to eradicating CML stem cells in the 55th ASH annual meeting is reviewed.

Objective: To investigate the expression of Cyclin G mRNA in leukemia patients and its clinical significance. Methods: RT-PCR was used to analyze the expression of Cyclin G mRNA in the mononuclear cells of 129 leukemia patients and 10 healthy controls. Results: (1) The expression of Cyclin G in acute leukemia (AL) and chronic leukemia (CL) patients was significantly higher than that in healthy controls (both P 50?109/L in AL and AML groups were significantly higher than those with WBC ≤ 50?109/L. (4) Fifty-three of the newly diagnosed cases were Cyclin G positive. The remission rate of patients with high Cyclin G expression(51.7%)was significantly lower than that with low Cyclin G expression(79.1%)(P

Objective To investigate the expression pattern of Nrf2 in the lung of septic rat and preliminary analysis of the role of Nrf2 in the development of sepsis. Methods Wistar rats were used in this study, it was divided into 4 groups, including normal control group, pure burn group, burn with staphylococcus sepsis group, burn with pseudomonas sepsis group. According to the different time intervals such as 2 hours, 8 hours, and 24 hours, it was divided into three sub-group after injection of bacteria. The expression of Nrf2 in the lung at different time intervals was determined. Results Nrf2 mRNA in the lungs of normal rats was high expression （74.0±7.0）, Nrf2 mRNA in the lungs of pure burns rats obviously down-regulated, respectively as 34.5±1.9,50.4±2.2,32.1±1.4, （t＝5.69～14.63,P＜0.01）. Nrf2 mRNA in burn sepsis caused by Staphylococcus aureus in lung tissue of rats down-regulated expression, respectively as 53.1±5.0,14.4±1.6,48.5±1.9,and reached peak at 8 h（t＝5.59～29.3,P＜0.01）. Pseudomonas aeruginosa burn sepsis didn't induced Nrf2 mRNA in the lung tissue, but it showed a downward trend at 2h（71.0±8.1,P＞0.05）and markedly reduced after 8, 24 h（24.8±2.1,4.1±2.0,t＝21.33,68.1,P＜0.01）.Conclusions The distribution of Nrf2 mainly localized in immune organ, and it directly took part in the post burn immune response.

AIM: To investigate the relationship between the expressions of DNA topoisomerase (Topo), glutathione S-transferases (GSTs) and chemotherapy response, prognosis in acute leukemia (AL). METHODS: Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) was used to detect the mRNA expression of TopoⅠ, Ⅱ?, Ⅱ? and GST?, ? from patients with AL. RESULTS: The results showed that the relative mRNA expression level of TopoⅡ?, TopoⅡ? and GST ? in AL group were significantly higher than that in normal subjects. GST?, however, was exactly reverse ( P

AIM:To study the effect of bcr - abl gene antisense phosphorothioate oligonucleotides(Aspo) on K562 cell line and explore its significance in chronic myelogencous leukemia (CML) gene therapy. METHODS: Cells were exposed to oligomers, observed by inverted microscope. Cells inhibitory rate were determined by 0.4% trypan blue exclusion. CFU - K562 were cultured in 0.8% methylcellulose. P210 was measured by flow cytometry. RESULTS:K562 cells were treated with Aspo, they still grew in clone state and show antisense sequence specific and dose dependent. When the concentration of Aspo was more than 5?mol/L, the growth of cells was inhibited and P210 was down regulated or completely suppressed, and the greatest growth inhibition was at 120h. There was significant inhibition of cell proliferation in a rang of cells number from 1 ? 10-4/mL to 5 ? 10-4/mL after treatment with 10?mol/L Aspo. b2a2 Aspo was also effect on K562 cells which expressing b3a2 mRNA. CONCLUSION: bcr - abl Aspo has a specific growth inhibition effect on K562 cells, and worths further study in CML gene therapy.

AIM:To investigate the effects of baicalin on proliferation inhibition and apoptosis induction in human Burkitt lymphoma cell line CA46 and to explore its underlying mechanisms.METHODS:CA46 cells were exposed to baicalin at different dosages and its proliferation inhibition was detected by MTT assay.The ability of baicalin to induce CA46 cell apoptosis was examined by Annexin V-FITC/PI double staining analysis,TUNEL labeling method and DNA fragmentation.The mRNA expressions of c-myc and bcl-2 were detected by RT-PCR,and the protein expressions of c-Myc,Bcl-2,caspase-3 precursor(procaspase-3) and poly ADP-ribose polymerase(PARP) were detected by Western blotting.RESULTS:Baicalin remarkably inhibited the CA46 cell proliferation,with an IC50 value of 10 ?mol/L.Apoptosis was remarkably induced by baicalin in a dose-dependent manner,and its earlier and later stages were detected by annexin V-FITC/PI double staining analysis,TUNEL labeling method and DNA fragmentation,respectively.Furthermore,RT-PCR showed that the mRNA expressions of c-myc and bcl-2 in treated CA46 cells decreased in a time-dependent manner.Western blotting showed that the protein expressions of c-Myc,Bcl-2,procaspase-3 and PARP(116 kD) in baicalin treated CA46 cells were down-regulated in a time-dependent manner,while the expression of PARP(85 kD) was up-regulated.CONCLUSION:Baicalin efficiently induces proliferation inhibition and apoptosis in CA46 cells,which may be related with the down-regulation of c-Myc and Bcl-2 expressions,as well as the up-regulation of caspase-3 activity.

AIM: To observe the effect of antisense bcl-2 oligodeoxynucleotides(AS-PS-ODN) on bcl-2 mRNA and protein expression, cell proliferation,viability and apoptosis in a small-cell lung cancer cell line NCI-H446. METHODS: Semi-quantitative RT-PCR was performed to detect the bcl-2 mRNA expression, the Bcl-2 protein was determined by immunocytochemistry and flow cytometry analysis, and the effect of bcl-2 AS-PS-ODN on cell proliferation, viability and apoptosis were investigated by colony assay , cell count, DNA content analysis and TUNEL. RESULTS: ① 1 ?mol/L bcl-2 AS-PS-ODN significantly down-regulated the expression of bcl-2 mRNA and protein. The inhibition rate of mRNA and protein were 69.5% and 62.7%, respectively. ② bcl-2 AS-PS-ODN decreased cell proliferation and viability , induced cell apoptosis.The apoptosis rate was 22.3%-32.7% in cells treated with 1?mol/L bcl-2 AS-PS-ODN. CONCLUSION: bcl-2 AS-PS-ODN down-regulated expression of bcl-2 mRNA and protein, inhibited cell proliferation and induced apoptosis in a small cell lung cancer cell line, NCI-H446.

Objective:To investigate the effect of SU11248 proliferation and apoptosis of multiple myeloma cell line U266 in vitro and analyze its mechanisms.Methods:Effect of SU11248 on proliferation of U266 cells was detected by MTT assay.The ability of SU11248 to induce apoptosis of U266 cells was examined by cell cycle analysis,TUNEL and DNA fragmentation.Expression of c-myc,hTERT,Bcl-2 and Bax mRNA in U266 cells was assessed by RT-PCR analysis.Results:The proliferation of U266 cells was inhibited by SU11248 in dose-and time-dependent manners (P

Objective To construct the eukaryotic expression plasmid containing human epidermal growth factor(hEGF) gene with signal peptide(SP).Methods After two pairs of primers were designed and synthesized,the cDNA fragment of hEGF and SP genes were amplified from total RNAs. The amplified cDNA fragments were cloned into pGEM-T vector.The expression plasmids were verified by double endonuclease digestion and DNA sequence analysis. Results With RT-PCR using two pairs of primers,two bands(about 90bp and 180bp) were obtained and confirmed as signal peptide and EGF cDNA fragment with electrophoresis analysis and DNA sequencing after cloned into pGEM-T vector.The SP and EGF cDNA fragments were inserted into plasmid pcDNA3.1(+).The bands of 240bp and 5.4kb were obtained and identified as the full length of SP-EGF cDNA fragment by DNA sequence analysis.Conclusion The eukaryotic expression plasmids containing hEGF gene is successfully constructed.

Objective To investigate DSA- estimated hepatic arterial hemodynamics of hepatocellular carcinoma (HCC) determined shortly after transcatheter arterial chemoembolization (TACE) plus sorafenib treatment. Methods The clinical data of thirty HCC patients treated with TACE were retrospectively analyzed. The patients were divided into study group (n = 13) and control group (n = 17). Patients in the study group received additional oral administration of 400mg sorafenib twice a day one week before or two weeks after TACE procedure, while patients in the control group received TACE only. The initial DSA images as well as the images obtained at three months after TACE were analyzed. With the help of Photoshop software, the grey gradient of the tumor staining was measured on the series dynamic DSA images, based on which the time- density curve of the tumor was drawn. The peak density value (PV), the time to reach the peak (TP) and the slope of the upslope (SU) were determined, and the results were compared between the two groups. Results Photoshop software was used to measure the grey density values of the tumor staining on DSA images. In the study group, the post- treatment PV was smaller than the pre- treatment one, which were (38.0 ± 14.6) and (46.7 ± 18.4) respectively, the difference between the two groups was statistically significant (P = 0.040). The post- treatment PV of the study group was also smaller than that of the post -treatment PV of the control group (54.4 ± 19.8), and the difference between the two was also statistically significant (P = 0.011). No significant differences in TP values and SU values existed between the two groups as well as between the pre - treatment and post - treatment ones in each group. Conclusion After TACE.