1.Expression and significance of FasL and FAP-1 in acute myeloid leukemia
Journal of Leukemia & Lymphoma 2009;18(8):481-483
Objective To explore the expression of the mRNA level of Fas (CD95) ligand/FasL and Fas-associated phosphatase-1/FAP-1 in acute myeloid leukemia. Methods The expression of FasL and FAP-1 were detected in 54 patients with AML and 10 normal subjects by semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR). β-actin used as internal reference. The changes of FasL was observed after induction chemotherapy in 16 AML patients. The expression of Fas was detected in 54 patients with AML by flow cytometry. Results The mRNA levels of FasL and in 54 patients were remarkably higher(P <0.05) than the normal controls. The rates of first complete remission(CR) were significantly higher in FasL(+) (77.78 %) than in FasL(-) cases(16.67 %) (P<0.01). The FasL level declined in 10 patients with good response to therapy (P<0.01), and high level in 6 nonresponding patients with poor response (P>0.05). The mRNA levels of FAP-1 in 54 patients was remarkly higher than that of the normal control. 8/29 cases in Fas-positive group were positive for FAP-1 mRNA expression. 19/25 cases in the Fas-refractory group didn't express FAP-1 mRNA. Conclusion The expression of FasL was high in AML. The rates of complete remission were high in FasL positive cases. The FasL level declined in patients with good response to therapy. The expression of FAP-1was partly expressed in AML. The expression of FAP-1 was less in Fas positive group.
2.Costimulatory molecules and allograft rejection
Cancer Research and Clinic 2006;0(12):-
Allograft rejection after xero organ transplantation rejection, especially acute rejection is still the major reason of failure and death. Active T cell play key roles in allograft rejection. It has been showed that the expressions of costimulatory molecules are associated with xero organ transplantation rejection. The pathways of CD28/CTLA-4 and CD40/CD40L are important costimulatory pathways that cause T cell activation.The article emphasizes on the role of CD28/CTLA-4-B7 pathway in allograft rejection.
3.The effect of allo-human bone marrow mesenchymal stem cells on interleukin-1, tumor necrosis factor-α, transforming grouth factor-β1 of patients with rheumatoid arthritis
Lihui MA ; Zhenhua QIAO ; Zhen YAO
Chinese Journal of Rheumatology 2011;15(9):615-618
ObjectiveTo study the effect of allo-human bone marrow mesenchymal stemcells (bMSCs) on the secretion of interleukin(IL)-1, tumor necrosis factor(TNF)-α and transforming grouth factor (TGF)-β of patients with rheumatoid arthritis (RA) in vitro. MethodsBMSCs were isolated from bone marrow of healthy volunteers and purified by density gradient centrifugation and cultured in vitro. The mononuclear cells from the peripheral blood of patients with RA and healthy controls were isolated respectively.bMSCs and mononuclear cells were co-cultured in vitro and the density of IL-1, TNF-α and TGF-β3 in the co-culture system were detected by ELISA. ANOVA and Pearson correlation were used for statistical analysis.ResultsMononuclear cells from peripheral blood of patients with RA were co-cultured with bMSCs for seven days. There were an decreased density ofIL-1[(38.4±0.5) vs(6.2±1.0) ng/L], TNF-α[(29.4±1.3) vs (4.6±1.2) ng/L]and an increased density of TGF-β[(2.6±1.0) vs (22.5±2.2) ng/L]in the co-culture system (P<0.05). But on the other hand, for healthy volunteers there were no significant change in the density of IL1[(4.4±1.1) ng/L]and TNF-α[(5.0±1.7) ng/L]in the coculture group, as compared with the mononuclear cell group[(4.4±1.3) vs(5.3±1.7) ng/L](P>0.05). There was an increased density of TGF-β in the coculture system[(4.8±1.4) vs(10.5±1.2) ng/L](P<0.05). IL-1 was positively correlated to TNF-αt (r=0.896,P=0.000), TNF-β1 was nagative correlation with 1L-1 and TNF-α (r=-0.356,P=0.019; r=-0.380,P=0.000).ConclusionHuman bone marrow MSCs have modulatory effects on main cytokines of patients with RA in vitro. bMSCs could down-regulate the levels of IL-1 and TNF, but up-regulate the density of TGF-β. These immune-modulatory effects are not MHC restricted. The results of this study have provided evidence for the development of effective therapy for RA.
4.Study of proliferation profile in cord blood CIK cells
Qiujuan ZHU ; Zhenhua QIAO ; Bo JIANG
Cancer Research and Clinic 1999;0(05):-
Objective To investigate the proliferation and the change of antigen on the cord blood CIK cells. Methods Cord blood CIK cells were generated by culture cord blood mononuclear cells in the presence of ?-IFN on day 0 and rhIL-2, antiCD3 MCAb on day 1. CIK cultures were analysed on different time point by FACS(Fluorescence activated cell sorter). The killing efficacy in tumor cell lines and primary leukemia cells were detected by MTT assay. Results After 2~3 weeks of culturing cord blood CIK cells expanded about 64.14?16 folds and CD+3 CD+56 cells which were the major cytolytic effector in CIK cells proliferated about 900 folds. Cord blood CIK cells have great cytotoxity against tumor cell lines as well as primary leukemia cells. Conclusions Cord blood MNCs can be cultured to CIK cells and the times of expansion are very high. Cord Blood CIK cells are highly efficient cytolytic effector cells.
5.Analysis of the infectious complications following nonmyeloablative allogeneic peripheral blood stem cell transplantation
Hongfeng GUO ; Zhenhua QIAO ; Liping SU
Chinese Journal of Organ Transplantation 2003;0(05):-
Objective To investigate the infectious complications after nonmyeloablative allogeneic peripheral blood stem cell transplantation (NASCT). Methods Ten patients with leukemia received NASCT from HLA-identical or 1-3 antigen mismatched sibling donors with conditioning regimens of Flu/Cy+BU+ALG. Five patients were acute leukemia (AL) and all of them is in complete remission (CR1). Cyclosporine in combination with methotrexate was administered for GVHD prophylaxis. Results WBC recovered to more than 0.5 ?10 9/L during postoperative 10 day to 38 day and platelet recovered to more than 20?10 9/L during postoperative 8 day to 16 day. Bacterial infections occurred in 3 patients (30 %) and cytomegalovirus (CMV) infections in 2 patients (20 %). Varicella zoster virus (VZV) infections occurred in one patient (10 %). No fungal infections were documented. No patients died as a result of infection . Conclusion NASCT is a safe and effective therapeutic method for leukemia. It reduces acute GVHD,regimen-related toxicity and early neutropenia associated with traditional allo-HSCT.
6.The distribution and effect of allogenic mesenchymal stem cells on joints of collagen induced arthritis rats
Lihui MA ; Zhenhua QIAO ; Xiaofeng LI
Chinese Journal of Rheumatology 2011;15(6):376-380,后插1
Objective To study the distribution of allogenic bone marrow-derived mesenchymal stem cells (BM-MSCs) on joints of collagen-induced arthritis (CIA) rats and to investigate their repair effects on joint damages. Methods Five Wistar rats were used for extraction of mesenchymal stem cells and 30 adult female Wistar rats were divided into 3 groups: the CIA rats group A (n=10), CIA rats group B (n=10) and normal rats control group C (n=10). BM-MSCs of Wistar rats were isolated, cultured in vitro routinely and the fourth passages was taken for identification of specific surface antigens by flow cytometry, then the cells were labeled with 5-bromodeoxyuridine (5-BrdU) in vitro. The models of CIA rats were established. 5-BrdU labeled BM-MSCs (1.0×107 cells/kg) were imfused from through tail vein to CIA rats group A and control group C. During the first 4 weeks after BM-MSCs transplantation, changes of general condition and left hind paw swelling were examined. At the fourth week, immunohistochemical examination of 5 -BrdU and osteoprotegerin (OPG) were performed to investigate BM-MSCs aggregation around the knee joints. The contribution of BM -MSCs to repairing of joint damages was identified. Comparisons between groups were performed by t-test. Results After BM-MSCs transplantation, left hindpaw swelling of group A were relieved compared with group B (P<0.05) and the mobility of the joints was significantly improved. At the fourth week, much more implanted cells (5-BrdU positive cells.) were detected in the damaged knee joints than those in normal knee joints. The average grey scale values on synovium of knee joints in the CIA group A (85±9) was significantly lower than that of the normal group C (110±6, P<0.05). At the same time, OPG expression was increased in damaged knee joints. The average grey scale values on synovium of knee joints in CIA group A (54±4) was significantly lower than that of the CIA group B (77±6, P<0.05). Conclusion The transplanted allogeneic bone marrow mesenchy-mal stem cells can migrate to sites of damaged tissue in arthritis. They can prevent tissue damage and repair the damaged joints tissue by increasing OPG expression. This study has provided some evidence for developing effective therapy for rheumatoid arthritis.
7.Drug analysis of Li zhenhua for the treatment of chronic atrophic gastritis based on the data mining
Lijun BAI ; Zhenhua LI ; Hua QIAO
International Journal of Traditional Chinese Medicine 2015;(2):161-164
Objective The method of data mining and sorting analysis was used to analyze and summarize the drug experience of Professor Li Zhenhua in the treatment of chronic atrophic gastritis (CAG). Methods Professor Li’s medical records of effective diagnosis and treatment of 139 CAG patients were collected. Find-Replace method of Microsoft office word 2007 was used to count the major syndromes and main prescription of CAG. SPSS statistical software was adopted to perform entry-analysis-descriptive statistics and data analysis of and main syndrome, main formula and frequency of administration so as to obtain the commonly used drugs, commonly used prescription and drug laws of CAG. Results Professor Li Zhenhua believed that the clinical syndromes of CAG included the disharmony of liver-stomach-spleen syndrome, the damp heat of spleen-stomach syndrome, the deficiency and damp heat of spleen syndrome, the liver depression and spleen deficiency syndrome, the deficiency of spleen-stomach syndrome, the liver and stomach yin deficiency and qi stagnation syndrome, the stagnant heat of liver-stomach syndrome and the blood stasis of stomach meridian syndrome;the commonly used drugs were:bupleurum, white peony root, orange peel, licorice, poria, skullcap, ginger, fried atractylodes, golden thread, prepared pinellia, licorice, lily, stir-baking Sanxian, nutgrass galingale rhizome, heterophylly falsesatarwort root, combined spicebush root, Chinese date, tangshen, immature orange fruit, prepared rhizome pinellize without adjuvant, and oyster shell..The commonly used prescriptions were: Xiaochaihu decoction, Sini powder, Chaihu-Guizhi-Longgu-Muli decoction, Chaihu-Shugan powder, Huanglian-Wendan decoction, Banxia-Xiexin decoction, Xiaoyao powder, Xiangsha-Liujunzi decoction. Conclusion Professor Li pay attention to treat spleen and stomach disease from liver by clearing heat and removing dampness from spleen and stomach. He used the dialectical methods like invigorating qi and strengthening the spleen, regulating qi digestion, activating blood flow to eliminating blood stasis.
8.Construction of human eukaryotic vector with expression of Survivin in K562 cells
Fang YE ; Lan ZHANG ; Tao YANG ; Zhenhua QIAO ; Linhua YANG
Journal of Leukemia & Lymphoma 2009;18(11):647-649
Objective To construct an eukaryotic vector with expression of human survivin gene with green fluorescent protein which is named pIRES2-EGFP/survivin and transfected into K562 cell line.Methods Using pDNR/Survivin plasmid as a template, the full length of survivin cDNA was amplified by PCR and subsequently cloned into T-A vector and then subcloned into pIRES2-EGFP vector. After identified by digestion of restrictive endonucleases, pIRES2-EGFP/survivin was further confirmed by sequencing. Then it was transfected into K562 cells with superfect reagents. The mRNA was isolated and survivin gene was detected by Western blotting. Results The exact sequences of pIRES2-EGFP/survivin vector were confirmed by digestion of restrictive endonucleases and sequencing. After transfection, the expressions of green fluorescent protein were present. The mRNA expression of survivin has been detected in transfected cells by RT-PCR. Conclusion The vector pIRES2-EGFP/survivin has been constructed and could express survivin gone in K562 cells successfully.
9.Isolation and culture of bone marrow mesenchymal stem cells and the study of their biological char-acteristics in patients with rheumatoid arthritis
Lihui MA ; Zhenhua QIAO ; Xiaofeng LI ; Liyun ZHANG
Chinese Journal of Rheumatology 2008;12(10):680-683,插2
Objective To isolate and culture bone marrow mesenchymal stem cells (BMSCs) from patients with rheumatoid arthritis (RA) and examine their biological characteristics. Methods MSCs were isolated from bone marrow of RA patients and purified by density gradient centrifugation and cultured in vitro. The morphology, immunophenotype, and proliferative; property of BMSC and colony forming unit-fibroblast (CFU-F) were measured and analyzed. Results The culture expanded cells from RA patients presented a typical fibroblast-like morphology. Ceils were positive for SH2 (CD105), CD71, and CD44, but negative for CD45. Their proliferative capacity and CFU-F number were similar to those of BMSCs from healthy donors. Conclusion In respect to morphology, immuno-phenotype, proliferative property and colony forming unit-fibroblast (CFU-F), MSCs from bone marrow of RA patients are not different from those of MSCs isolated from bone marrow of normal donors. MSCs from the bone marrow of RA patients have the potential for clinical application.
10.Humanistic spirit cultivation of the medical students during the clinical internship
Yajun QIAO ; Heqing HUANG ; Kangning CHEN ; Yihong WU ; Zhenhua ZHOU
Chinese Journal of Medical Education Research 2012;11(1):93-95
The clinical internship is an important transitional period for humanistic spirit cultivation of the medical students.But in this period there are some problems exposed.For example,there is the lack of transitional link in management; the clinical instruction doctors lack educational consciousness or guidance ability,the medical students pay little attention to enhancing the individual humanistic spirit level and there is a gap between theory and practice of humanistic spirit for them.Strengthening training for students,attaching importance to training,selection and incentive of clinical instruction doctors and revising handbook of clinical internship will help to solve the problems.