1.NS-398, a selective cyclooxygenase-2 inhibitor, can down-regulate the expression of P-glycoprotein in the gastric cancer cell
Gen LIN ; Yuli CHEN ; Zhizhe CHEN
Chinese Journal of Digestion 2001;0(09):-
Objective To investigate the effect of NS-398, a selective cyclooxygenase-2 (COX-2) inhibitor, on the expression of P-glycoprotein(P-gp) in a gastric cancer cell line SGC-7901. Methods SGC-7901 cells were treated with NS-398 in different concentrations (0, 10 ?mol/L and 100 ?mol/L) respectively. Prostaglandin E2(PGE2) was detected by ELISA. Twenty four hs and 48 hs later, the expression of mdr1 mRNA were detected by RT-PCR. P-gp protein expression in SGC-7901 cells was detected by immunocytochemical technique after NS-398 treatment.Results NS-398 can inhibit the expression of PGE2 in a dose-dependent manner(P
2.Application of HLA-DRB、DPB1 genotyping by PCR-SSP、RFLP for allogeneic bone marrow transplantation
Xianghua UN ; Zhizhe CHEN ; Minhui LIN
Chinese Journal of Immunology 2000;0(08):-
Objective:To investigate the feasibility of clinical application of HLA-DRB,DPB1 genotyping by PCR-SSP,RFLP for alloge-neic bone marrow transplantation(Allo-BMT) .Methods: Corresponding to each of the DRB alleles, nineteen different pairs of sequence specific primers were used to PCR amplify them from twenty samples in both recipients and donors and their amplified products were directly analyzed on agarose gel. At the same time, a pair of primer was used to PCR amplify the second exon of DPB1 and the restriction fragment length polymorphisms were analyzed on PAGE after their PCR products were separately digested by ten endonucleases. Results: Each DRB was typed in according to its clear and visualized electrophoresis band; the DPB1 genotype was also determined by analysis of all codes representing polymorphism fragment band with a computer. Four couples of recipient and donor were matched. Conclusion:PCR-SSP and RFLP are rapid, accurate and reliable genotyping approaches for Allo-BMT.
3.Study of differentiation from CML into dendritic cell by calcium ionorphore
Hairong ZHOU ; Junmin CHEN ; Zhizhe CHEN ; Chenqing ZHANG
Chinese Journal of Immunology 2001;0(10):-
Objective:To explore the methods and condition in which calcium ionorphore(CI) induces the CML cells to differentiate into dendritic cells(DCs).Methods:Mononuclear cells were separated from peripheral blood or bone marrow of CML patients whose WBC counts were more than 30?30~9 L~ -1 when samples were collected,then lymphocytes and monocytes were discarded by pouring out supernatant twice at different culture time point. Slightly adherent cells were cultured in RPMI1640 containing 10%FCS(Fetal calf serum),with or without CI(375 ng/ml) and GM-CSF(200 ng/ml) at 37℃,5%CO_2 humidified atmosphere for 96 h. To evaluate the effect of CI on inducing CML cells to differentiate into DCs,the phenotype of these cells were analyzed by flow cytometry and the morphology change was observed under inverted microscope and electron microscope. Better condition was also explored for DCs differentiation from CML cells under the effect of CI.Results:After 96 h of culture with CI and GM-CSF,the CML cells acquired morphology of mature DCs and significantly up-regulation of CD80,CD86,CD40,CD86 and HLA-DR.Conclusion:CML cells might acquire typical morphology and immunological phenotype of mature DCs when being cultured with CI and GM-CSF.
4.Cyclooxygenase-2 inhibitors enhance the antileukemia activity of STI571
Xianguo WU ; Zhizhe CHEN ; Yaping JIN ; Ting YANG ; Yueling YANG
Chinese Journal of Pathophysiology 2000;0(08):-
AIM: To investigate whether celecoxib,a cyclooxygenase-2(COX-2) inhibitor,potentiates the anti-leukemia activity of STI571 in K562 cells.METHODS: K562 cells were treated with STI571,celecoxib or combination of both at different concentrations in suspension culture.Cell proliferation was documented by MTT assay,and cell apoptosis was determined by flow cytometry and morphology.Meanwhile,RT-PCR was applied to analyze the probable mechanism underlying the effects of the drugs.RESULTS: The combination of STI571 and celecoxib dramatically suppressed the proliferation of K562 cells,in which 0.25 ?mol/L STI571 and 40.0 ?mol/L celecoxib enhanced the inhibiting rate to 76.1%?1.6%.Furthermore,the combining administration of drugs significantly promoted the apoptosis induced by STI571,which showed characteristic changes of morphologic features and increase in sub-G_1 cells.By using RT-PCR technique,the expression of COX-2 had no decline by single administration of celecoxib or STI571.However,a progressive down-regulation was caused by coadministration of two drugs.In contrast with COX-2,the expression of VEGF had no changes at any time.CONCLUSION: The administration of celecoxib alone only inhibits the proliferation of K562 cells.Combination treatment with STI571 and celecoxib promotes the apoptosis induced by STI571.
5.Dynamic change of the reticulin fibres and collagen fibres in vitro long-term bone marrow culture of acute myeloid leukemia
Yanjuan LIN ; Lianhuang LU ; Zhizhe CHEN ; Juemin LI ; Pingrong ZHANG
Chinese Journal of Pathophysiology 1986;0(01):-
AIM and METHOD: The relationship between the evolution of the reticulin fibres(RF) or the collagen fibres(CF) and the growing of hematopoietic cells in long-term bone marrow culture (LTBMC) from 15 paticnts with acute myeloid leukemia (AML) and form 6 normal subjects was observed by inverted microscope, Gomoris staining and Massons staining were used. RESULTS: (1)The amount of RF contents of 8 AMLs,with self- maintained(AMLsm) in the 1~8 weeks-old-culture was significant less than that of normal control and 7 AMLs,without self-maintained(AMLnsm) ( P
6.Cytokine production and hematopoiesis-supportive function of human umbilical cord mesenchymal stem cells
Lulu Lü ; Yongping SONG ; Baijun FANG ; Yanli ZHANG ; Yufu LI ; Langhui ZHANG ; Zhizhe CHEN
Journal of Leukemia & Lymphoma 2008;17(6):404-407
Objective To investigate the cytokine spectrum and henlatopoiesis-supportive function of umbilical cord derived mesdnchymal stem cells(UC-MSC),and compare with those of normal adult bone marrow derived mesenchymal stem cells(BM-MSC).Methods The mRNA of cytokine production of UC-MSC and BM-MSC were determined by reverse transcriptasc polymerase chain reaction(RT-PCR)analysis.To evaluate hematopoiesis supporting activity,cord blood(CB)CD+34 cells were co-cultured with UC-MSC or BM-MSC.Colony-forming cells(CFC)were determined after 5 weeks of culture.Results RT-PCR assay showed that UC-MSC had a cytokine spectrum very similar to that of BM-MSC.including expression of the mRNA ofstem cell factor,leukemia inhibitor factor,macrophage colony stimulating factor,Flt3-ligand,interleukin-6,vascular endothelial growth factor and stromal derived factor-1.but UC-MCS additionally expressed mRNA of granulocyte macrophage and granulocyte colony-stimulating factors.After co-culture with CD+34 cord blood cells for 5 weeks,no significant difference in CFC was observed between the CD+34 cells/UC-MSC and CD+34 cells/BM-MSC co-cultures (P>0.05). Conclusion The cytokine spectrum and hematopoiesis-supponive function of UC-MSC ale similar with that of BM-MSC.
7.Up-expression of IL-6 and down-expression of TNFalpha may be involved in the regulation of apoptosis induced by antisense bcl-2 oligodeoxynucleotides.
Xinji CHEN ; Jianda HU ; Zhizhe CHEN ; Lianhuang LU
Chinese Journal of Medical Genetics 2002;19(6):495-498
OBJECTIVETo investigate the regulation mechanism of apoptosis induced by the antisense bcl-2 treatment.
METHODSDNA content analysis and terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL) were adopted to detect apoptosis. Semi-quantitative reverse transcription-PCR was performed to detect the mRNA expression of bcl-2 c-myc survivin bax s100A(2) TNFalpha TGFbeta(1) and IL-6 in the small-cell lung cancer cell line NCI-H446 treated with antisense bcl-2 oligodeoxynucliotide.
RESULTSbcl-2 AS-PS-ODN treatment could induce apoptosis, accompanied with 72.71% up-regulation of IL-6 and 65.90% down-regulation of TNFalpha, whereas little or no effect was seen on c-myc survivin bax s100A(2) and TGFbeta(1).
CONCLUSIONIL-6 and TNFalpha may be involved in the regulation of apoptosis induced by antisense bcl-2 treatment.
Apoptosis ; drug effects ; genetics ; Chemotactic Factors ; genetics ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; In Situ Nick-End Labeling ; Inhibitor of Apoptosis Proteins ; Interleukin-6 ; genetics ; Microtubule-Associated Proteins ; genetics ; Neoplasm Proteins ; Oligonucleotides, Antisense ; pharmacology ; Proto-Oncogene Proteins ; genetics ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; Proto-Oncogene Proteins c-myc ; genetics ; RNA, Messenger ; drug effects ; genetics ; metabolism ; S100 Proteins ; genetics ; Transforming Growth Factor beta ; genetics ; Transforming Growth Factor beta1 ; Tumor Cells, Cultured ; drug effects ; Tumor Necrosis Factor-alpha ; genetics ; bcl-2-Associated X Protein
8. Distribution and drug resistance of pathogens of blood stream infection in patients with hematological malignancies after chemotherapy
Shaozhen CHEN ; Kangni LIN ; Min XIAO ; Xiaofeng LUO ; Qian LI ; Jinhua REN ; Ruoyao HUANG ; Minmin CHEN ; Issa Hajji ALLY ; Zhizhe CHEN ; Ting YANG ; Jianda HU
Chinese Journal of Hematology 2017;38(11):951-955
Objective:
To investigate the distribution and resistance of pathogens isolated from blood cultures in patients with hematological malignancies after chemotherapy in Union Hospital of Fujian Medical University so as to understand the real situation of blood stream infection (BSI) and provide the basis for rational use of antibiotics in clinic.
Methods:
The data of 657 strains isolated from blood culture specimens of patients with hematological malignancies from January 2013 to December 2016 were collected analyzed.
Results:
A total of 657 cases of blood culture positive bacterial strains were included in the study, involving 410 cases (62.4%) with single Gram-negative bacteria (G- bacteria) , 163 cases (24.8%) with single Gram-positive bacteria (G+ bacteria) , 50 cases (7.6%) with single fungi. The most common 5 isolates in blood culture were