P Zou

Tongji Hospital, Wu-han-shih, Hubei, China

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Publications (17)0.9 Total impact

  • W Cao, Y Zhang, D Zhang, P Zou
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    ABSTRACT: To explore the effect of NF-kappa B on bcl-x gene transcription in extended drug resistance leukemia cell line HL-60/E6, drug-resistant subline HL-60/E6 was derived by intermittently exposing HL-60 cells to 6 ng/ml epirubicin. Indirect immunofluorescence was used to demonstrate the location of NF-kappa B-RelA in HL-60/E6 cells. FCM analysis and RT-PCR were used to detect the efficiency of liposome-mediated ODN transfection and the change of bcl-XL mRNA levels after 5 mumol/L phosphorothioate (PS)-derivatized antisense (AS) oligodeoxynucleotide (ODN) directed to RelA was transferred into HL-60/E6 cells. The results showed that RelA remained persistently active and located at the nuclei of HL-60/E6 cells, but in the cytoplasm of HL-60 cells, the efficiency of liposome-mediated ODN transfection was significantly higher than that of null ODN (P < 0.01 in 4 h, 6 h, 12 h, 24 h). Exposure of HL-60/E6 cells to 5 mumol/L AS-PS-ODN directed to RelA led to a maximal 40% decline of bcl-XL mRNA levels within 8 h. The inhibition rate of bcl-XL mRNA was (15 +/- 1.79)%, (28 +/- 2.34)%, (40 +/- 3.47)%, (20 +/- 1.54)%, in 4 h, 6 h, 8 h, 15 h, respectively, but it was less than 15% in control group. It was concluded that NF-kappa B was involved in regulating bcl-x transcription. It was suggested that NF-kappa B was an important factor for drug resistance in leukemia cells.
    Journal of Tongji Medical University = Tong ji yi ke da xue xue bao 02/2001; 21(1):32-4.
  • W Li, Z Chen, Z Liu, P Zou
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    ABSTRACT: In order to study the significance of flow cytometry immunophenotyping in the diagnosis of acute leukemia, CD45/SSC gating multiparameter flow cytometry (FCM) was utilized to analyze the immunophenotypes of 139 cases of acute leukemia. 139 cases of acute leukemia were enrolled in our hospital from April 1998 to April 2000. Morphological analysis and FCM immunophenotypic tests were conducted on all cases. Our results showed that CD45/SSC gating multiparameter flow cytometry immunophenotyping could reflect the origin of leukemic cells specifically. It is one of the important methods for the diagnosis of ALL, AML, and HAL. CD45/SSC gating multiparameter FCM analysis is a good technique for immunophenotyping. FCM immunophenotypic analysis can help improve the diagnosis and classification of acute leukemia, and extend the use of FCM in clinical practice.
    Journal of Tongji Medical University = Tong ji yi ke da xue xue bao 02/2001; 21(3):209-11.
  • L Liu, P Zou, R Guo, J Xiao, Z Xu
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    ABSTRACT: The bioactivities of culture supernatants from retroviral packaging cells carrying the mouse Fas ligand (mFasL) gene was investigated. FasLcDNA was cloned into PLXIN with an internal ribosome entry site to link two cistrons through gene recombination technology, PLXIN and the recombinant vector PLFIN were separately transfected into PA317 retrovirus packing cell line by lipofectamine 2000, and the resistant clones were selected with G418 selective medium. The integration of genome DNA was assayed by genomic DNA PCR. NIH3T3 cells were transduced by the culture supernatants from PA317 carrying the mFasLcDNA gene, and were selected with G418 selective medium, so as to select the PLFIN-PA317 clone capable of producing higher titer of supernatants. The levels of mFasL protein on NIH3T3 cells membrane were assayed by flow cytometry (FCM). The biological activity of mFasL on NIH3T3 cells membrane was investigated by the inducing apoptosis of Fas+ Yac-1 cells co-cultured with NIH3T3 cells expressing Fas ligand. To explore the direct mFasL cytotoxicity of culture supernatants from retroviral packaging cells carrying the mFasL gene, the culture supernatants from PLFIN-PA317 and PLXIN-PA317 were separately co-cultured with Yac-1 cells in parallel. The recombinant PLFIN was successfully constructed. The highest titer of supernatants from twelve resistant clones was 8.5 x 10(5) colony-forming-unit (CFU)/ml. The NIH3T3 cells transfected by above supernatants had a higher level of mFasL (53.81 +/- 6.9%), and significantly induced the apoptosis of Fas+ Yac-1 cells (56.78 +/- 4.5%), as both were cocultured for 5 h at 1:1 ratio, whereas it is 7.08 +/- 3.4% in control group (P < 0.01). Supernatant from PLFIN-PA317 could also directly induce the apoptosis of Yac-1 within 5 h of incubation. Thus, the culture supernatants from PLFIN-PA317 possessed both infectivity and cytotoxicity of mFasL.
    Journal of Tongji Medical University = Tong ji yi ke da xue xue bao 01/2001; 21(3):215-8.
  • P Zou, Z Liu, J Xiao
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    ABSTRACT: The mechanism of chemotherapeutic drug-induced apoptosis in leukaemic cells was studied to further investigate whether Fas/FasL system was involved in apoptosis induced by chemotherapeutic drugs and assess their effects when used in combination with soluble FasL (sFasL). The expression of Fas on human leukaemic cell lines K562, HL-60 and U937 treated with daunorubicin (DNR) or cytosine arabinoside (Ara-C) was detected by using flow cytometry. The activities of sFasL, DNR and Ara-C inducing apoptosis of leukaemic cells, in the absence or presence of neutralizing anti-Fas IgG antibody, were detected by using flow cytometry and TUNEL. The results showed that flow cytometric profiles of K562, HL-60 and U937 cells treated with DNR or Ara-C failed to show any significant increase in Fas expression over 18 h (P > 0.05). Anti-Fas monoclonal antibody (IgG) could not block the apoptosis in leukaemic cells induced by DNR or Ara-C, but could block the apoptosis induced by sFasL. A role of sFasL in a cytotoxic synergistic effect when used in combination with chemotherapeutic drugs was revealed. It was concluded that chemotherapeutic drug-induced apoptosis in human leukaemic cells (UG37, HL-60) is independent of the Fas/FasL system, but combination of sFasL and drug treatment produces a synergistic cytotoxic effect on human leukaemic cells.
    Journal of Tongji Medical University = Tong ji yi ke da xue xue bao 01/2001; 21(3):212-4.
  • L Wang, P Zou, Y You
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    ABSTRACT: To study the differences in homing potential between bone marrow cells and umbilical blood cells, CD34 positive cells were obtained from bone marrow (BM) and umbilical blood (UB) by the direct cell separation with domestic immunomagnetic beads. The expression of the two adhesion molecules CD11a/CD18 and CD44 were examined. After separation, CD34 positive cells accounted for 51%-82% of the harvested cells and dye-resistance rate was 82%-88%. The expression of CD11a/CD18 and CD44 on the surfaces of UB cells was 49.6% +/- 10.2% and 37.7% +/- 10.3% respectively. On BM cells they were 50.2% +/- 6.2% and 34% +/- 13.3% respectively. There were no significant differences in the expression of these two molecules. It was concluded that the cell separation method with domestic immunomagnetic beads was effective and the stem cells from UB could serve as an alternative source for transplantation.
    Journal of Tongji Medical University = Tong ji yi ke da xue xue bao 02/2000; 20(2):132-3.
  • X Liu, Z Tang, P Zou
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    ABSTRACT: The rearrangement of Bcl-1 gene (Bcl-1/IgH rearrangement) and expression of cyclin D1 in multiple myeloma (MM) precursor cells were studied and the role of cyclin D1 in the pathogenesis of MM was investigated. The BCL-1 rearrangement and cyclin D1 protein expression in 15 cases of MM were detected. By using hemi-nested polymerase chain reaction (PCR) the genomic DNA from fresh peripheral blood and bone marrow was amplified and the expression of cyclin D1 in the smears was detected by using immunohistochemical method. Ten volunteer with normal bone marrow served as control group. The results showed Bcl-1 rearrangement was detectable in 3/15 (20%) MM patients and cyclin D, expression in 4/15 (27%) MM patients. Bcl-1 rearrangement and cyclin D1 protein expression were also detected in MM precursor cells. No overexpression of cyclin D1 or the rearrangement of the Bcl-1 gene was found in the 10 volunteers. It was concluded that Bcl-1 rearrangement and cyclin D1 protein overexpression were detected in MM precursor cells, speculating that overexpression of cyclin D1 protein may play an initial (critical) role in the pathogenesis of MM.
    Journal of Tongji Medical University = Tong ji yi ke da xue xue bao 02/2000; 20(2):128-31, 136.
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    ABSTRACT: To study the possibility of dauricine (Dau) inhibiting redistribution of platelet membrane glycoprotein IV (GPIV) and release of intracellular alpha-granule thrombospondin (TSP) on platelet activation. Using the flow cytometric assay of washed platelet to record expression of GPIV and release of TSP induced by thrombin. Dau did not affect GPIV and TSP on resting platelet membrane but inhibited redistribution of GPIV to the platelet surface and TSP release on activated platelet. There was a marked positive correlation between changes of GPIV and TSP (r = 0.511, P < 0.01). The inhibitory effect of Dau appeared not to be Ca2+ concentration-dependent. Dau inhibited redistribution of GPIV and release of intracellular alpha-granule thrombospondin induced by thrombin.
    Zhongguo yao li xue bao = Acta pharmacologica Sinica 07/1999; 20(6):533-6.
  • Z Liu, Y You, Z Chen, P Zou
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    ABSTRACT: The changes of cell surface markers before and after activation by IL-2 were detected by flow cytometry (FCM) to establish a more convenient and precise criterion for the judgment of the activation of bone marrow. By using the measurement of the release of lactate dehydrogenase (LDH) the cytotoxicity of mononuclear cells (MNCs) from bone marrow, activated or inactivated, on tumor cell line K562 was evaluated, and at the same time the changes of surface markers on MNCs before and after activation were examined by using FCM. The results showed that the cytotoxicity of MNCs from bone marrow activated by IL-2 on tumor cell line K562 was increased obviously and the number of CD25+ and CD70+ positive cells in bone marrow MNCs was higher than before activation. The enhanced cytotoxicity of MNCs on tumor cell line K562 was synchronous with the increase of the number of CD25+ and CD70+ positive cells in 48 to 72 h. It is more direct, simple and precise to demonstrate the activation of IL-2 on bone marrow by detecting the changes of the amount of the CD25+ and CD70+ positive cells in bone marrow by FCM.
    Journal of Tongji Medical University = Tong ji yi ke da xue xue bao 02/1999; 19(4):264-6.
  • S Liu, X Liu, P Zou, S Song, B Wang
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    ABSTRACT: An immunohistochemical study of T lymphocyte subsets on frozen substituted plastic embedding bone marrow sections obtained from 10 patients with myelodysplastic syndrome (MDS) was presented. The results of qualitative and quantitative immunohistochemical analysis are as follows: (1) Labile antigens of T lymphocytes were well preserved, thus allowing analysis of distribution of T lymphocyte subsets in situ: (2) the average number of T3, T4 and T8 lymphocyte of the diffuse infiltrate was about 2%, 0.4%, 0.5%, respectively, of all nucleated cells in bone marrow, and T4/T8 of T cells were below 1.0 in patients with MDS; (3) there were cases of RAS showing T lymphocyte aggregation in bone marrow, but no patient exhibited progressive refractory anemia with excess of blasts (RAEB) and RAEB in transformation (RAEBT). These findings indicated that the immunological abnormalities are of importance in the evaluation of pathogenesis and prognosis of MDS.
    Journal of Tongji Medical University = Tong ji yi ke da xue xue bao 02/1999; 19(3):198-202.
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    ABSTRACT: The clonal growth of human acute leukemia cell line (K562) and acute myeloid leukemia cells in the serum-free culture (SFC) was studied in order to establish a SFC system which could replace the effects of serum by using semi-solid methylcellulose culture technique. Our results showed that the clonal growth of K562 cells in semi-solid culture was dependent on exogenous serum. The K562 could be grown in SFC supplemented with 4 major replacing substances. The multifactor and multilevel orthogonal experiment demonstrated that the colony formation was statistically influenced by the 4 replacing substances at various concentrations (P < 0.01). Among them, bovine serum albumin had greatest effect on clonal growth of K562 cells with the optimal concentration being 15 mg/L, followed by transferring, cholesterol and insulin with their optimal concentrations being of 150 mg/L, 7.8 mg/L and 7.0 mg/L respectively. SFC system was formed with the 4 substances at their optimal concentrations. Colony formation of the blast cells in 10 patients with acute myeloid leukemia was observed in this SFC system. There was a heterogeneity of acute myeloid leukemia cells among the 10 patients in response to the growth substances. In SFC system, there was a linear relationship between the number of the clonal formation and the count of the added cells, indicating the colony growth of the cells. Primary acute leukemia cells maintained in SFC system in 10 cases could completely form clones. The colony formation number in some cases in SFC system was more than that of the serum-containing culture. The SFC system could partially replace the serum for study of the clonal formation of human leukemia cells.
    Journal of Tongji Medical University = Tong ji yi ke da xue xue bao 02/1999; 19(3):190-3.
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    ABSTRACT: The multidrug resistance P-glycoprotein (P-gp) expression and function in hematopoietic stem/progenitor cells were studied to investigate whether the inhibition of hematopoietic cell P-gp function by multidrug resistance reversal agent increases the cytotoxicity of chemotherapy drugs on the hematopoietic cells. The expression of P-gp on the surface of CD34+ cells from healthy human marrow was examined by flow cytometry. The multidrug resistance reversal agent MS-209 was used to measure the effects of MS-209 on the Rhodamin-123 uptaking of CD34+ hematopoietic cells. By using methylcellulose semi-solid culture, normal human granulocyte-macrophage clonal formation unit (CFU-GM) was cultured. The changes in CFU-GM inhibitory rate caused by daunorubicin were determined in the presence or absence of MS-209. The results showed that the P-gp expression rate of bone marrow CD34+ cells was 13.3%. MS-209 obviously increased the Rhodamin-123 uptake of CD34+ positive cells. The mean inhibitory rate of daunorubicin for CFU-GM was 29.6%, but it was increased to 43.3% in the presence of MS-209 with the difference being significant (P < 0.05). It was concluded that hematopoietic cells expressed P-gp protein and possessed active function. MS-209 could inhibit the membrane efflux pump and increase the cytotoxicity of chemotherapy drugs to the clonal growth of hematopoetic stem cells, suggesting the side effects of these drugs on the hematopoietic system should be taken into consideration in the clinical use.
    Journal of Tongji Medical University = Tong ji yi ke da xue xue bao 01/1999; 19(4):260-3.
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    ABSTRACT: To investigate the activity of platelet in patients with high level of low density lipoproteins (LDL) and the effect of LDL on platelet glycoproteins (GP). Platelet glycoproteins, whose platelet were activated or unactivated, were measured by flow cytometry. The amount of the platelet membrane GP II b/III a in patients was not significantly different from that of the control when platelet was unactivated (P > 0.05); when platelet was activated by adenosine diphosphate (ADP), the amount of patients' platelet GP II b/III a was increased markedly in comparison with that in the control (P < 0.01). Having been preincubated with LDL, the platelets were activated by ADP and the amounts of GP II b/III a of patients group and the control were all increased obviously as compared with those in the platelets which were not incubated with LDL (P < 0.01; P < 0.01). There was no significant difference of the amount of granule membrane protein-140 (GMP-140) between patients and the control when platelet was activated by thrombin (P > 0.05). Having been preincubated with LDL, platelet was activated by thrombin, the amounts of GMP-140 on patients' and control platelet were all increased markedly (P < 0.01, P < 0.01). The activity of platelet in patients with high level of LDL is increased significantly. LDL can increase the expression of platelet glycoproteins.
    Chinese medical journal 10/1998; 111(10):910-2. · 0.90 Impact Factor
  • P Zou, H Lu, J Xiang
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    ABSTRACT: We have shown previously that high-efficient gene transfer can be attained in primary hematopoietic cells using liposome-mediated gene transfer strategy. In order to examine the stability of gene expression mediated by this gene transduction protocol, we observed the expression of marker gene in vivo by using bone marrow transplantation (BMT) to engraft lethally irradiated mouse with the genetically modified hematopoietic cells. The results showed that the mouse transplanted with appropriated number of transduced cells remained alive and healthy. The PCR analysis and G418 selection of the spleen colonies and bone marrow cells isolated from lethally irradiated animals 15 days and 30 days after injection of genetically modified bone marrow cells showed that the progeny cells of the transduced hematopoietic stem cells still contained and expressed the transduced genes, suggesting that the hematopoietic system is at least partially re-constructed by the stem cells with marker gene and that the stable expression of foreign genes in vivo can be attained by using this easy and harmless transduction protocol. These findings provide experimental basis for clinician to further investigate the biology of marrow reconstruction and the mechanism of leukemia relapse after BMT.
    Journal of Tongji Medical University = Tong ji yi ke da xue xue bao 02/1998; 18(1):46-8, 53.
  • H Lu, P Zou, C Li
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    ABSTRACT: To explore the efficiency of liposome-mediated gene transfer into bone marrow cells and the stability of its expression. Two marker genes (Neo and Lac Z) mediated by liposome were co-transferred into bone marrow cells. Following G418 screening,the positive cells were enriched and expanded in vitro. The liposome-mediated gene transfer rate in bone marrow cells was 13.33 +/- 2.68%, and after G418 screening, it was 46.06 +/- 3.47%. The blue colouring rates with X-gal staining of these cells were not significantly different before and after expansion. Liposome-mediated gene transfer in bone marrow cells is highly efficient and the expression of the transferred gene is stable.
    Zhonghua xue ye xue za zhi = Zhonghua xueyexue zazhi 09/1997; 18(8):422-4.
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    ABSTRACT: To explore the regulatory effects of granulocyte-macrophage colony stimulating factor (GM-CSF) on the apoptosis of leukemic cells induced by Vp16. High expression retrovirus vector, N2A/CMV/GM-CSF, was constructed and transferred into human leukemic cell line HL-60, and Vp16 (final concentration 10microg/ml) was administered to transferred or nontransferred HL-60 cells. After Vp16 treatment, characteristic changes for apoptosis emerged in HL-60 cells. However, these findings did not emerge in GM-CSF gene transferred HL-60 cells. GM-CSF can suppress the leukemic cell apoptosis induced by Vp16.
    Zhonghua xue ye xue za zhi = Zhonghua xueyexue zazhi 07/1997; 18(6):299-301.
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    ABSTRACT: To study the relationship between the expression of adhesion molecules CD49d (VLA-4) and CD11a (LFA-1) and the invasiveness of acute myeloid leukemia (AML) cells. Peripheral blood and/or bone marrow samples from 50 AML patients were investigated by APAAP and Western blotting method. Extramedullary invasion developed in 32 of 50 patients (64%). The expression of CD49d and CD11a in the invasive group was much higher than that in the non-invasive group (P<0.005), while the difference between the leukemic cells from bone marrow and peripheral blood for CD49d/CD11a expression was not significant. AML cells might adhere to and get through vascular endothelium by CD49d/VCAM-1 and CD11a/ICAM-1 adhesion mechanism, and the expressions of CD49d and CD11a were not critically responsible for the release of leukemic cells from bone marrow.
    Zhonghua xue ye xue za zhi = Zhonghua xueyexue zazhi 02/1997; 18(1):29-31.
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    ABSTRACT: The redistribution of platelet membrane glycoprotein IV (GPIV) and the release of intracellular alpha-granule thrombospondin (TSP) were examined and the inhibition of beta-thromboglobulin (beta-TG) and platelet factor 4 (PF4) in patients with chronic myelogenous leukemia (CML) was observed and quantitation of beta-TG and PF4 in sera was conducted. GPIV in inactive platelet from CML was 36080 +/- 17010 molecules/platelet as compared with 13190 +/- 4810 from the controls (P < 0.01). No abnormality was found in the distribution of platelet membrane GPIb and GPIIb/IIIa (P > 0.05). The GPIV redistribution on active platelet membrane induced thrombin (IU/ml) from CML and healthy donors was 44320 +/- 32310 and 22800 +/- 12700 molecules/platelet respectively (P < 0.01). The difference in the release of intracellular alpha-granule TSP between CML and the control group was not found (P > 0.05). There was no direct correlation between GPIV expression and TSP binding after platelet activation. The high levels of beta-TG and PF4 in sera inhibited release of intracellular alpha-granule TSP in vitro. These results indicate that the abnormality of platelet membrane GPIV is a common marker in CML, therefore the specific increase of platelet GPIV in patients with CML may be a useful tool for the diagnosis and monitoring of the platelet dysfunction. The release of internal TSP pools is hindered by either beta-TG or PF4 in sera.
    Journal of Tongji Medical University = Tong ji yi ke da xue xue bao 01/1997; 17(1):21-4.