1.Construction and expression of human anti-HBs-IFN fusion gene
Le JIANG ; Jinqi YAN ; Bingran GUO ; Jie REN ; Jiyun YU
Medical Journal of Chinese People's Liberation Army 1981;0(04):-
Objective To construct eukaryotic expression plasmid pEE14.1-dsFv?pr+,and detect the expression of the recombined gene in eukaryotic CHO-K1 cells.Methods The cationic DNA fragment was cloned into the 3' of VH gene by overlapping extension PCR,and the 6?His tab was inserted to the 3' of VL and human IFN-? gene by the same way.The above mentioned recombinant VH and VL genes were inserted into a pCI-GPI vector first,and then cloned into the pEE14.1 vector to construct the recombinant plasmid pEE14.1-dsFv?pr+.Finally,the recombinant plasmid was transfected into the CHO-K1 cells by LipofectamineTM 2000,and the expression was detected by RT-PCR,ELISA and Western blotting.Results The enzyme digestion and sequencing analysis showed that the recombinant plasmid was successfully constructed.RT-PCR showed that only the cells with transfected plasmid can generate the specific 1700bp fragment.ELISA analysis showed that the production of IFN-?expressed in the supernatant of transfected cells was about 1.1ng/ml.Also,Western blotting could reveal the characteristic band of HBsAg dsFv?pr+ protein.Conclusion The antibody targeting to human IFN-?genes has been successfully expressed in a single open reading frame.Changing the electricity of the antibody may provide the necessary condition for the study of the a new type of anti-HBV drug in nanoscale in the future.
2.Clinical analysis of 20 patients with angioimmunoblastic T-cell lymphoma
Zhi GUO ; Xuepeng HE ; Kai YANG ; Xiaodong LIU ; Peng CHEN ; Bing LIU ; Dan LIU ; Bingran WANG ; Huiren CHEN
Journal of Leukemia & Lymphoma 2012;21(1):42-46
Objective To improve the diagnosis efficiency of patients with angioimmunoblastic T-cell lymphoma (AILT)by analyzing the clinical characteristics and curative effect of AILT. Methods Retrospective studies were used on clinicopathological features,immunophenotypes,treatment and survival of 20 angioimmunoblastic T-cell lymphoma patients,who were collected between January 2005 and January 2010 of Beijing Military General Hospital. Results In the 20 patients receiving chemotherapy,the median age was 55.9 years old.All of the 20 had lymph nodes and 11 of whom were accompanied with B group of symptoms,which were confirmed by lymph node biopsy and T cell antigen CD3, CD45Ro positive expression in all the patients. ALL of the patients received the CHOP regimen and combining with other treatment such as DICE,ESHAP, Hyper-CVAD or autologous hematopoietic stem cell transplantation. A follow-up by 5 to 69 months showed that median survival was 20.2 (5-69) months and 1,2,and 3-year disease free survival (DFS) were 67 %,33 %,and 11% respectively.8 patients of the group (40 %) with a sustained remission (CCS) respectively survived 63,47,27,24,24,12,5,1 months so far.Conclusion Most patients were older with many complications during chemotherapy, which resulted in easy relapse, and even the symptoms were alleviated at the early stage of the chemotherapy. IPI prognostic index is more important with long-term survival in AITL.Therefore further studies are required to improve the outcome.