1.Expression of human IL-24 gene in CHO cells and its anti-tumor effect in vitro
Jingcheng MIAO ; Xiongyan CHEN ; Weihua SHENG ; Yufeng XIE ; Jicheng YANG
Chinese Pharmacological Bulletin 1986;0(05):-
Aim To construct the eukaryotic expression vector of hIL-24 cDNA,and express it in CHO cells and detect its anti-tumor effect of recombinant hIL-24 protein.Methods Constructed pcDNA3-hIL-24 was identified by endonucleases digestion & PCR.The recombinant expression plasmids were transfected into CHO cells,human hIL-24 expressed in CHO cells was detected with RT-PCR.The apoptosis-inducing activities of recombinant protein hIL-24 was tested by MTT assay,Hoechst& FCM assay,and the expression of IL-6 and IFN-? from PBMC induced by rhIL-24 was tested by ELISA.Results The eukaryotic expression vector pcDNA3-hIL-24 was constructed correctly.Stable expression of human IL-24 in CHO cells was identified with RT-PCR.The apoptosis of A549 cells induced by hIL-24 was proved by Hoechst & FCM assay,and the expression of IL-6 and IFN-? from PBMC induced by rhIL-24 was identified with ELISA.Conclusion The successful stable expression & experimental study of apoptosis effect of human IL-24 gene lay the foundation for the further study of molecular mechanism of hIL-24 on anti-tumors and potential application.
2.Anti-thrombopoietin antibodies in systemic lupus erythematosus with thrombocytopenia
Xiaodan WU ; Xiongyan LUO ; Long CHEN ; Jing ZHANG ; Tong WU ; Minghui YANG ; Bin ZHOU
Chinese Journal of Rheumatology 2012;16(11):745-748
Objective To investigate the existence and significance of circulating autoantibodies to thrombopoietin (TPO) in sera from patients with systemic lupus erythematosus (SLE).Methods Fifty-six consecutive patients with SLE,twenty patients with idiopathic thrombocytopenic purpura (ITP),and twenty normal individuals were involved in this study.The characteristics of all patients with SLE were analyzed.Antibodies to TPO were detected using an enzyme-linked immunosorbent assay (ELISA).For normal distribution count data,x2 test or Fisher exact test was used,t test was used for measurement data,and Wilcoxon's rank test for non-normally distributed data which was represented by M(Q).Results A higher frequency of antibodies to TPO were observed in SLE patients than those in healthy controls (39.3% vs 0,x2=11.058,P=0.001).Moreover,anti-TPO antibodies were detected in 15 (57.7%) of 26 SLE patients with thrombocytopenia,compared with that in 7 (23.3%,x2=6.894,P=0.009) of 30 patients without thrombocytopenia.Furthermore,the patients with antibodies to TPO exhibited more severe thrombocytopenia (t=3.010,P=0.004).Finally,anti-TPO antibodies seemed more likely to occur in patients with arthritis (x2=5.959,P=0.015),anti-dsDNA antibodies (x2=5.959,P=0.015).Conclusion The high incidence of antibodies to TPO in SLE patients with thrombocytopenia suggests that anti-TPO antibodies might play a vital role in SLE patients developing thrombocytopenia.Thus,there might be a clinical value by detecting anti-TPO antibodies in SLE patients with thrombocytopenia.
3.The association of vitamin D receptor gene Apa Ⅰ and Bsm Ⅰ polymorphism with systemic lupus erythematosus
Xiongyan LUO ; Lijun WU ; Long CHEN ; Minghui YANG ; Tao LIAO ; Ningtao LIU ; Banjiang KUER ; Chuanmei XIE ; Rangeng SHI ; Zhong TANG ; Yan ZHAO ; Xiaofeng ZENG ; Guohua YUAN
Chinese Journal of Internal Medicine 2012;51(2):131-135
ObjectiveTo determine the distribution of vitamin D receptor (VDR) gene Apa Ⅰ and Bsm Ⅰ polymorphism in systemic lupus erythematosus (SLE) and the association with SLE in Chinese Han patients.MethodsGenomic DNA from 244 Chinese SLE patients and 162 sex and ethnically matched controls were typed for VDR Apa Ⅰand Bsm Ⅰpolymorphism combination by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP). Clinical characteristics were analyzed between different Apa ] and Bsm Ⅰ genotypes.ResultsThere was no significant difference between the distribution frequencies of allelic gene A and a in SLE patients and the controls,but the distribution frequency of genotypes heterozygote Aa in SLE patients was higher than that in the controls ( 38.9% vs 22.2%,x2 =12.442,P =0.000).There was no significant difference between the distribution frequency of allelic gene and genotypes of Bsm Ⅰin SLE patients and the controls ( P > 0.05 ).However,there was significant difference between the distribution frequencies of Apa Ⅰ and Bsm Ⅰ genotypes combination in SLE patients and the controls (x2 =18.226,P =0.006).The distribution frequency of genotypes Aa-bb in SLE patients was higher than that in the controls ( 32.4% vs 17.9%,x2 =10.449 P =0.001 ),while the distribution frequency of genotypes Aa-bb in SLE patients was lower than that in the controls (30.3% vs 42.0%,x2 =5.808,P =0.016). Furthermore,analyzing the effect of VDR Apa Ⅰand Bsm Ⅰ polymorphism combination to the symptoms of SLE,significant difference was observed in SLE patients carrying Aa-bb genotypes involved in serositis ( P =0.003 ),hematological system disorder ( P =0.021 ),and anti-Sm antibodies ( P =0.01 ) compared with other genotypes.ConclusionThere is significant association between Apa Ⅰ and Bsm Ⅰ gene polymorphism Aa-bb genotypes and the incidence of SLE in the Han population of China,and genotype Aa-bb is more involved in serositis,hematological system disorder and has a positive effect on production of antibodies.