1.Clinical analysis of decitabine combine with different regimens in the treatment of elderly patients with acute myeloid leukemia
Kai YANG ; Huiren CHEN ; Xuepeng HE ; Jixing LOU ; Zhi GUO ; Yuan ZHANG ; Peng CHEN
Journal of Leukemia & Lymphoma 2014;23(8):484-487
Objective To investigate the therapeutic effects and adverse reactions of decitabine combined with IA or CAG regimen in the treatment of elderly patients with acute myeloid leukemia.Methods A retrospective analysis was made to observe the therapeutic effects and adverse reactions of 47 elderly patients with acute myeloid leukemia,who were divided into DAC+IA group (17 cases) and DAC+CAG group (28 cases) according to the different chemotherapy.Results In DAC+IA group,the rate of complete remission was 29.4 % (5/17),the rate of partial remission was 35.3 % (6/17),the effective rate was 64.7 % (11/17).In DAC+CAG group,the rate of complete remission was 26.7 % (8/30),the rate of partial remission was 30.0 % (9/30),the effective rate was 56.7 % (17/30),the difference between the two groups was not statistically significant (x2 =0.227,P =0.716).In DAC+IA group the median remission time and the median suvival time were 4.0 and 8.1 months,respectively.And they were 4.0 and 8.1 months,respectively,in DAC+CAG group.No significant difference was showed between the two groups (P value was 0.835,0.266,respectively).Conclusions Compared with decitabine combined with CAG regimen,decitabine combined with IA regimen has similar effect and can be well tolerated.Accordingly,decitabine combined with IA regimen can be used as first-line treatment for elderly patients with acute myeloid leukemia.
2.Prokaryotic Expression and Potential Application of the Truncated PCV-2 Capsid Protein
Zhongzi LOU ; Zhiyong LI ; Gang WANG ; Jianqiang LI ; Xi LAN ; Xuerui LI ; Xiangping YIN ; Jixing LIU ; Sidang LIU
Virologica Sinica 2010;25(2):86-97
Three pairs of specific primers were designed to amplify the F2-1, F2-2 and XF2-2 truncated sequences of ORF2 which encodes the capsid protein of porcine circovirus type 2 (PCV-2). The F2-1 sequence had most of the NLS region of ORF2, but the F2-2 and XF2-2 genes had the NLS region deleted. Truncated genes were subcloned into pET-32a(+) vectors to construct recombinant fusion expression vectors. The vectors were then transformed into Rosetta(DE3) E. coli and expressed by induction of IPTG. Expressed proteins were detected by western blotting and ELISA. The protein with best immunoreactivity was confirmed and selected, then utilized to inoculate SPF rabbits to prepare polyclonal antibodies. The protein and prepared polyclonal antibody were utilized to detect sera samples against PCV-2 from Shandong province and PCV-2 particles in PK-15 cells. In our study, three recombinant fusion proteins were successfully obtained, and the molecular weights of fusion proteins were 35.9 kDa, 33.6 kDa and 38.6 kDa respectively detected by SDS-PAGE. All of the proteins showed positive reaction with anti-PCV-2 antisera, and His-XF2-2 showed better immunoreactivity than the others. The protein of His-XF2-2 was coated as antigen in ELISA to detect the seroprevalence of PCV-2 in certain districts of Shandong province, the seropositivity rate was 27.7 % (73/264). Specific fluorescence and positive signals for PCV-2 could be detected in PK-15 cells inoculated with PCV-2 with the participation of prepared antibodies against His-XF2-2 in IFA and IPMA. Experimental results indicated that the truncated PCV-2 ORF2 gene containing most of the NLS region was successfully expressed in E. coli, and His-XF2-2 was demonstrated to have better immunoreactivity with anti-PCV-2 antisera than the other two fusion proteins. His-XF2-2 and prepared polyclonal antibodies against it had a satisfactory capability in detecting PCV-2 infection.
3.Cloning the Structure Genes and Expression the N Gene of Porcine Epidemic Diarrhea Virus DX
Jianqiang LI ; Jixing LIU ; Xi LAN ; Jie CHENG ; Run WU ; Zhongzi LOU ; Xiangping YIN ; Xuerui LI ; Baoyu LI ; Bin YANG ; Zhiyong LI
Virologica Sinica 2009;24(3):179-186
The structure genes spike (S), nucleocapsid (N), membrane (M), small membrane (sM) of a porcine epidemic diarrhea virus (PEDV) strain DX isolated in Gansu province, North-west of China, were cloned, sequenced and compared with published sequences of PEDV strains. The nucleotide sequences encoding the entire S, sM, M and N genes open reading frame (ORF) of DX were 4 152, 231, 681 and 1 326 bases long respectively. There were transcription regulatory sequences (TRSs) upstream of the initiator ATG of the S, N and M genes. The amino acids sequences of S, M and N contained 30, 3 and 7 potential asparagine (N)-linked glycosylation sites. Homologous analysis and phylogenetic trees showed that DX had the closest relationship with strains LJB/06, JS-2004-2Z and CH/HLJH/06 that were also isolated from China and indicated the prevalence of some PEDV isolates in China were widespread since the JS-2004-2Z strain originated from the south of the China, and LJB/06 and CH/HLJH/06 were isolated from northeast China. The N gene was cloned using two primers which contained Nco I and BamH I restriction enzyme sites and subcloned into expression vector pET30a. The recombinant plasmid was then transformed into E.coli Rossta. SDS-PAGE showed there was a protein of about 55kDa as expected and Western blot indicated the N protein had biological activity.