1.Cytokine expressions in peripheral blood mononuclear cells during graft-versus-host disease after allogeneic hematopoietic stem cell transplantation
Fang YE ; Zhenhua QIAO ; Tao YANG ; Kunping GUAN ; Cunjiu GUO ; Linhua YANG
Journal of Leukemia & Lymphoma 2008;17(3):195-199
Objective To investigate the relationship between cytokines and human graft-versus-host disease (GVHD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT). Methods In 21 patients undergoing allo-HSCT,the plasma concentrations of cytokines[soluble interleukin 2 receptor(sIL-2R), interferon-gama (IFN-γ), transforming growth factor-betal (TGF-β1)] were measured by using sandwich enzyme-linked immunological assay (ELISA) and the gene expressions of three cytokines were analysed by using semi-quantitate reverse transcriptase-polymerase chain reaction(RT-PCR). Results The concentrations and gene expressions of sIL-2R and IFN-γin the patients with GVHD were significantly higher than those without GVHD (P <0.01), and they were higher in the patients with aGVHD than with cGVHD and without GVHD(P <0.05); the levels of TGF-β1 in the patients with GVHD were significantly declined(P <0.01), but in those without aGVHD were obviously increased(P <0.05). After effective treatment, unnormal sIL-2R, IFN-γand TGF-β1 expressions recovered to the levels before transplantation. A multivariate COX analysis showed sIL-2R and TGF-β1 are independent prognostic factors for GVHD (P<0.001). Conclusion Monitoring the changes of sIL-2R, IFN-γand TGF-β1 expression levels (especially sIL-2R and TGF-β1) might provide predictive markers for GVHD after allo-HSCT. The sensitivity between RT-PCR and ELISA for detecting cytokines expressions had no difference.
2.Effect of rhDCN on proliferation of leukemic cell line K562
Gang JING ; Yu ZHANG ; Cunjiu GUO ; Aiping ZHU ; Guiqin WANG ; Yanhong WANG
Journal of Leukemia & Lymphoma 2012;21(2):83-86
ObjectiveTo investigate the anti-tumorigenesis function of rhDCN on the leukemia K562 cells in vitro and analyze the possible mechanism.Methods Exponential phase of K562 cells were transfected with pcDNA3.1(+)-DCN,and PBS,liposome alone,and pcDNA3.1(+) vector were as control groups.Morphology change of K562 cells was detected by Wright stain,and cell proliferation activity was detected by MTT. Cell cycle and apoptosis of K562 cells were assessed by FCM. The expression of apoptosis-related protein,including bcl-xl,Mcl-1 and Bax were detected by Western blot.ResultsWright stain showed that typical apoptotic morphology of K562 cells were observed in DCN transfected group.There were no morphological changes of apoptosis in other groups. MTT method results showed that proliferation inhibition rate of the transfected cells [24 h (16.14±1.08) %,48 h (14.07±1.01) %,72 h (20.29±1.19) %]was higher than that of the other control groups (P < 0.05).FCM results showed that the apoptosis index (20.15±1.31) %of the DCN transfected group was higher than that of the other groups (P < 0.05),and most of cells arrested in the G0/G1 phase (51.15±0.57) % (P < 0.05).Western blot results showed the expression levels of Bax were increased while bcl-xl and Mcl-1 were decreased in pcDNA3.1(+)-DCN/K562 group.ConclusionrhDCN can inhibit the growth of K562 cells and induce the apoptosis.The effect of DCN on bcl-xl,Mcl-1,Bax may play a role in its mechanism.
3.Significance of hematology analyzer scatter chart and alarm information in the diagnosis of hematological malignancies
Na ZHANG ; Fanggang REN ; Cunjiu GUO
Journal of Leukemia & Lymphoma 2018;27(9):517-520
Objective To study the role of hematology analyzer in the preliminary diagnosis of hematological malignancies. Methods The blood routine results of 121 patients with initial hematological malignancies in the Second Hospital of Shanxi Medical University from October 2016 to February 2017 were analyzed by differential blood count (DIFF) scatter chart and alarm information of hematology analyzer SYSMEX XE-2100. Results Among 121 patients with hematological malignancies, 96 cases (79.3 %) had abnormality of DIFF scatter chart. The DIFF scatter chart of acute lymphoblastic leukemia, chronic lymphocytic leukemia, acute promyelocytic leukemia, and chronic myeloid leukemia had their own distinct characteristics, but the features of other types were not obvious. According to blood smear, blood cells analysis alarmed that the coincidence rates of primitive cells, immature granulocytes, nucleated erythroblasts and atypical lymphocytes were 87.7 % (50/57), 76.3 % (42/55), 33.3 % (19/57), 43.1 % (31/72) respectively. Conclusion The DIFF scatter chart of hematology analyzer and alarm information play very important roles in the diagnosis of hematological malignancies.
4.Prokaryotic expression of coxsackievirus group A type 16 VP1 protein
Zhihong WANG ; Yingying DING ; Jiaojiao FENG ; Lianqing LI ; Wei PAN ; Cunjiu GUO
China Modern Doctor 2014;(14):69-72,75
Objective To construct the recombinant plasmid of coxsackievirus group A type 16 (CA16)VP1,express in E.coli and identify expressed product. Methods The CA16 VP1 gene was amplified by PCR,The constructed recombi-nant plasmid pET21b-CA16 VP1 was transformed into E.coli BL21(DE3) for expession and purification,expressed and purified product was identified by SDS-PAGE,Indirect ELISA tested Antigenicity and validity of recombinant proteins CA16 VP1. Results The coxsackievirus group A type 16 VP1 protein was expressed in peokaryotic cells and purified, which showed specific reaction with mice antiserum against CA16 virus,high reactivity with part serums of the elderly population in Taiyuan city. Conclusion CA16 VP1 protein was successfully expressed, had good Antigenicity and va-lidity,which will lay a foundation for study on the CA16 diagnostic kits.