检索结果-南通市图书馆

Mitochondria-dependent apoptosis induced by a novel amphipathic photochemotherapeutic agent ZnPcS2P2 in HL60 cells

在线全文

学校读者我要写书评

暂无评论

Acta Pharmacologica Sinica 2005年第9期26卷 1138-1144页

作者： Hui-fang HUANG Yuan-zhong CHEN~2 Yong WU Fujian institute of hematology,Union Hospital,Fujian Medical University,Fuzhou 350001,China Fujian institute of hematology Union Hospital Fujian Medical University

Aim:To investigate the mechanism underlying the killing effects of a novelamphipathic photosensitizer,disulfonated diphthalimidomethyl phthalocyaninezinc(ZnPcS2P2),mediated photodynamic therapy(ZnPc-PDT)in human myelog-enous leukemia HL60 ***:After incubation for 5 h with 0.5 μmol/LZnPcS2P2,the HL60 cells were exposed to a light source of 670 nm ***,the cells were detected at different time intervals after *** char-acteristics of apoptosis were detected by observation of ultrastructure assay,DNA fragmentation assay and terminal deoxynucleotidyl transferase deoxyuridinenick-end labeling method(TUNEL).Mitochondria-dependent apoptosis was de-termined by the detection of mitochondrial membrane potential(Δψm),activitiesof caspase family protease and of caspase-3,cytosol cytochrome *** Bcl-2and Bax were detected by immunoblot ***:Evident characteristicsof apoptosis were observed post-ZnPc-PDT with ultrastructure assay,DNA frag-mentation assay and TUNEL *** assay showed that apoptotic ratesin the cells collected from 6 h,12 h and 24 h after PDT were 9.6%,24.4%,and 33.0%,***60 cells underwent mitochondria-dependent apoptosis as a re-sult of cytochrome c release from mitochondria into cytosol accompanied by areduction of Δψ*** activities of caspase family protease and of caspase-3were ***,ZnPc-PDT could remarkably down-regulate the Bcl-2pro-apoptotic protein and up-regulate the anti-apoptotic Bax ***:ZnPc-PDT could induce mitochondria-dependent apoptosis in HL60 cells.

关键词： photodynamic therapy zinc phthalocyanine apoptosis cytochrome c caspases HL60 cells

Recombinant fibronectin polypeptide antagonizes hepatic failure induced by endotoxin in mice

在线全文

同方期刊数据库

学校读者我要写书评

暂无评论

Acta Pharmacologica Sinica 2004年第6期25卷 81-86页

作者： Yong WU, Yuan-zhong CHEN2, Hui-fang HUANG, Ping CHEN Fujian institute of hematology, Union Hospital, Fujian Medical University, Fuzhou 350001,China Fujian institute of hematology Union Hospital Fujian Medical University Fuzhou 350001 China

AIM: To study the preventive effect of recombinant human fibronectin (rhFN) polypeptide on hepatic failure in- duced by endotoxin in mice. METHODS: A cDNA fragment coding for Ile1363-Tyr1725 of human FN was inserted into the PinPoint Xa-3 plasmid, and the constructed plasmid was transformed into E coli BL21 (DE3) cells, and then the expression of rhFN polypeptide in DE3 cells was identified through SDS-PAGE. The target protein from the supernatant of bacteria lysate was purified through biotin-affinity chromatography. The bioactivity of the purified rhFN polypeptide was determined with cell adhesiveactivity. The survival rate was observed in endotoxemia mice injected with rhFN polypeptide. The tissue damage in hepatocyte was detected using histology, ultrastructure, and DNA fragmentation assay. RESULTS: The expression of rhFN polypeptide reached approximately 20 % of the total cellular protein. The adhesion ability of rhFN polypeptide was concentration-dependent. The value of EC50 was 0.8 nmol/L. The survival rate of endotoxemia mice sensitized by D-galactosamine (D-GalN) was 60 % in rhFN polypeptide treated group, while that of endotoxemia mice sensitized by D-GalN was 20 % in the control group (P<0.01). Histopathology showed that less necrosis occurred on the hepatocyte of endotoxemia mice in- jected with rhFN polypeptide compared with saline control. Ultrastructure and DNA fragmentation assay showed that no apoptotic hepatocyte was observed in the liver of rhFN-treated endotoxemia mice. CONCLUSION: Re- combinant fibronectin polypeptide antagonizes hepatic failure induced by endotoxin in mice.

关键词： fibronectins recombinant fusion proteins lipopolysaccharides acetylgalactosamine

Immunohistochem ical Study of T Lym phocyte Subsets onFrozen Substituted Plastic Em bedding Section of the BoneMarrow from Patients with Myelodysplastic Syndrom e andIts ClinicalIm plication

在线全文

同方期刊数据库

学校读者我要写书评

暂无评论

Journal of Huazhong University of Science and Technology(Medical Sciences) 1999年第3期19卷 198-202页

作者： LIU Shangqing , LIU Xiufang , ZOU Ping , SONG Shanjun , WANG Bianming institute of hematology, Xiehe Hospital, Tongji Medical University, Wuhan 430022 institute of hematology Xiehe Hospital Tongji Medical University Wuhan 430022

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 T 3, T 4 and T 8 lymphocyte of the diffuse infiltrate was about 2 %, 0.4 %, 0.5 %, respectively, of all nucleated cells in bone marrow, and T 4/T 8 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.

关键词： immunohistochemistry plastic embedding of bone marrow distribution of T cell subsets in situ myelodysplastic syndrome

Clonal Growth of Hum an Acute Leukem ia Cells in Serum -free Methylcellulose Medium

在线全文

学校读者我要写书评

暂无评论

Journal of Huazhong University of Science and Technology(Medical Sciences) 1999年第3期19卷 190-193页

作者： CHEN Zhichao , ZOU Ping , YOU Yong , LIU Zhongping , XIANG Jianping , YU Dongjiao institute of hematology, Tongji Medical University, Wuhan 430030 institute of hematology Tongji Medical University Wuhan 430030

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.

关键词： leukemia cells serum free culture

Viewpoints on the proinflammation state of leukemia

在线全文

学校读者我要写书评

暂无评论

Chinese Science Bulletin 2003年第22期48卷 2519-2522页

作者： WU Kefu & MA Xiaotong National Laboratory of Experimental hematology, institute of hematology, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin 300020, China Correspondence should be addressed to Wu Kefu (e-mail: kfwu@ ***) National Laboratory of Experimental hematology Institute of Hematology Chinese Academy of Medical Sciences & Peking Union Medical College Tianjin China

Proinflammation represents a pathophysiological state on the early stage of a number of diseases, especially the infectious and immunological ones. In recent years, proinflammation has attracted much attention, and the term 損roinflammation factors?appears frequently in the literature. While investigating leukemia and leukemic cells from the angle of 損roinflammation state? we got some intriguing findings, e.g. we detected the significantly elevated expression of proinflammation factor IL-18 in patients with acute myeloid leukemia (AML), which could up-regulate matrix metalloproteinases (MMP) and specific tissue inhibitors (TIMPs). The increased MMP may play a role in the aggressiveness of myeloid leukemic cells, and be associated with a poor prognosis. This phenomenon reflects an ignored aspect of leukemia. Investigations from the angle of 損roinflammation state?have broaden the fields of tumor and leukemia study.

关键词：白血病生理学肿瘤细胞生长机制肿瘤治疗学

Inhibition of Fas-mediated Apoptosis in Yac-1 Cell via Anti-Fas Ribozyme

在线全文

学校读者我要写书评

暂无评论

Acta Biochimica et Biophysica Sinica 2004年第3期36卷 199-205页

作者： Min ZHANG, Fang LIU, Wei HE, Yong YOU, Ping ZOU*, and Ling-Bo LIU institute of hematology Disease, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China institute of hematology Disease Union Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430022 China

To detect a new and more effective way against apoptosis mouse lymphomatic cell line- Yac-1 in which fas gene was expressed highly was used as a model for studying the effects of anti-Fas ribozyme on Fas-mediated apoptosis. A hammerhead ribozyme gene targeting the fas mRNA was synthesized and its in vitro transcription vector was constructed, which was transfected into Yac-1 cells using electroporation. Rz596 expression was detected using RT-PCR, and Fas expression in Yac-1 cells was detected using RT-PCR, Western blot and flow cytometry. After treated with anti-Fas antibody (JO2), Yac-1 cell viability was measured with MTT assay, caspase-3 proteolytic activity was detected, and cell apoptosis was measured according to annexin V apoptosis detecting kit. Anti-Fas ribozyme could cleave fas mRNA efficiently in vivo and in vitro. Fas expression in Yac-1 cells transfected with anti-Fas ribozyme was decreased remarkably and correlated with resistance to Fas-mediated apoptosis as determined by flow cytometry and caspase-3 proteolytic activity. Anti-Fas ribozyme was detected in cells transfected with pU6-RZ596 and pU6-dRZ596 and could remarkably decrease the Fas expression in Yac-1 cells, which made Yac-1 cells get rid of Fas-mediated apoptosis. Because of wide expression of fas in organs and tissues, our research was very useful for studying the inhibition of apoptosis of many organs and tissues in the future.

关键词： transplantation immunology ribozyme Fas Yac-1 apoptosis

The Com prehensive Evaluation on Four Indices of Drug Re-sistance in Acute Myeloid Leukem ia

在线全文

同方期刊数据库

学校读者我要写书评

暂无评论

Journal of Huazhong University of Science and Technology(Medical Sciences) 1999年第3期19卷 194-197页

作者： CHEN Yan , HE Mingsheng , XIANG Zhifu , WU Yudan, YUE Beibei , YU Dongjiao, LI Huiyu institute of hematology, Xiehe Hospital, Tongji Medical University, Wuhan, 430022 institute of hematology Xiehe Hospital Tongji Medical University Wuhan 430022

To study sensitivity of drug resistance indexes and resistance manner in acute myeloid leukemia (AML), MTT drug sensitivity, growth types of CFU L in vitro , Bcl 2 antigen and Bcl 2/Bax ratio and intracellular fluorescence intensity of daunorubicin (DNR) were determined. In 62 cases of AML, the positive coincidence rate was 73 % with MTT test and the negative coincidence rate was 70 %. In 3 commonly used drugs, if one drug showed sensitivity, the coincidence remission rate reached 71 %. In 51 cases of AML, there were 31 patients in the group of complete remission (CR), in which CFU L of 29 patients showed independent growth. CFU L of 2 patients showed no growth. However, there were 20 patients in the group of non remission (NR), in which CFU L of 14 patients showed independent growth. CFU L of 6 patients showed non growth pattern. Statistical analysis showed significant difference ( P <0.05). In 32 cases of AML, the expression rate of Bcl 2 was 59.55 %±19.56 % in drug sensitive group, and one was 77.36 %±11.91 % in drug resistant group, respectively ( P <0.05). At the same time, the ratio of Bcl 2/Bax was 7.50±5.04 in drug sensitive group and one was 14.32±8.99 in drug resistant group, respectively ( P <0.05). In 15 case of clinically drug resistant AML, the fluorescence histogram of DNR showed left shift of main peak (LSMP) in 12 patients. They were diagnosed as classical drug resistance. Meanwhile, 1 patient showed right shift of main peak (RSMP) in 3 patients. They were diagnosed as re growth drug resistance. It is concluded that MTT and CFU L might be used for prediction of drug sensitivity or resistance when patients were on treatment. Bcl 2 and ratio of Bcl 2/Bax might be associated with the prognosis. DNR histogram could be employed for identify the pattern drug resistance. The strength and weakness of these techniques were discussed.

关键词： acute myeloid leukemia drug resistance curative effect

The Protein Expression of Bcl-2, Bax, Fas/Apo-1 in Acute Myeloid Leukemia

在线全文

学校读者我要写书评

暂无评论

Journal of Huazhong University of Science and Technology(Medical Sciences) 1998年第1期18卷 42-45页

作者：周剑锋陈燕李崇渔刘文励 Departmentofhematology TongjiHospitalTongjiMedicalUniversityWuhan.

The protein expression of bcl-2, bax, Fas/Apo-1 in 19 cases of acute myeloid leukemia (AML) were investigated by Western blot and APAAp techniques. High expression of bcl-2 protein was found in most of the AML cases,and some of the cases expressed Fas/Apo-1 and bax. High expression of bcl-2 protein was associated with a bad clinical prognosis and a poor response after intensive chemotherapy. Bax seemed to coexpress with bcl-2 and so appeared to be a bad prognostic factor instead of a good one. The expression of Fas/Apo-1 was inversely correlated with hcf-2 and seemed to be a good prognostic factor which may reflect the relative homeostasis of apoptotic pathway. It is concluded that apoptosis-induced pathways in AML often exhibit disturbance features. Coregulation of bcl-2, Bax and Fas/Apo-1 genes formed the apoptosis-induced pathway,which is the biological factor affecting response to chemotherapy.

关键词： apoptosis bcl-2 Fas/Apo-1 bax AML

Experimental studies of the effects of ZnPcS2P2-based-photodynamic therapy on bone marrow purging

在线全文

学校读者我要写书评

暂无评论

中华医学杂志(英文版) 2005年第2期 19-24页

作者： HUANG Hui fang, CHEN Yuan zhong and WU Yong Fujian institute of hematology, Union Hospital, Fujian Medical University, Fuzhou 350001, China

Background An effective purging technique plays an important role in autologous hematopoietic stem cells transplantation Photodynamic therapy (PDT) provides a novel approach for this purpose This study dealt with the purging effects of disulfodiphthalimidomethyl phthalolcyanine zinc (ZnPcS2P2)based photodynamic therapy (ZnPcPDT) Methods Fluorescence intensity of cell extracts was measured using a fluoresence spectrophotometry The proliferative potency of K562 cells and HL60 cells was detected using MTT colorimetric assay, Typan blue dye exclusion method, colony formation test The proliferative potency of normal hematopoietic cells was evaluated using mixture colonyforming unit (CFUMix), granulocytemacrophage colonyforming unit (CFUGM), and erythrocyte colonyforming unit (CFUE) assays K562 cells were mixed with normal mononuclear cells (MNCs) at ratios of 1∶100 and 1∶1000 for creating the model of simulated remission bone marrow Colony formation test and nestedRTPCR were carried out to detect the residual K562 cells in cell mixture Results After a 5hour incubation with ZnPcS2P2, the content of ZnPcS2P2 in normal MNCs was the lowest value At the same time, the content in K562 cells and HL60 cells was very high Therefore, the time point was selected as the optimal one for irradiating the cell suspensions ZnPcPDT could significantly kill proliferative K562 cells and HL60 cells in a dosedependent manner At the concentration of 10 μg/ml, the inhibitory rate of ZnPcPDT on the colony formation was 100% for K562 cells, 897% for HL60 cells 025 μg/ml ZnPcPDT could completely photoinactivate residual K562 cells in the simulated remission bone marrow Under an identical condition, the inhibitory rates of CFUMix, CFUGM, CFUE were 180%, 186%, and 178% respectively Conclusion ZnPcPDT appears to be a promising approach for bone marrow purging

关键词： photodynamic therapy · zinc phthalocyanine · bone marrow purging · leukemia