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91.
Differential etoposide sensitivity of cells deficient in the Ku and DNA- PKcs components of the DNA-dependent protein kinase 总被引:3,自引:0,他引:3
Etoposides block cell division by interfering with the action of
topoisomerase II, leaving enzyme-DNA double-strand breaks. We found that
certain components of the trimeric DNA-dependent protein kinase influence
cell survival following etoposide damage. Interestingly, either Ku70- or
Ku80-deficient cell lines, but not mutant cell lines of the DNA-PK
catalytic sub-unit (DNA-PKcs), were found to be hypersensitive to the
effects of etoposide VP16. Ku70- and Ku80- deficient cells can be
complemented to an etoposide resistant phenotype by introducing wildtype
Ku70 or Ku80 cDNAs. Mutational analysis of introduced Ku70 cDNAs into
murine embryonic stem cells deleted for Ku70 (-/-) showed that mutants
where heterodimerization and DNA binding functions of Ku were disrupted,
also blocked the restoration of etoposide resistance. In contrast with the
differential etoposide sensitivity of DNA-PK mutants, both Ku- and
DNA-PKcs-deficient cell lines showed G2 ionizing radiation-induced delays,
a cell cycle phase where topoisomerase II function is critical. Thus, the
topoisomerase II cleaved complexes may be an example of DNA lesions
requiring the Ku heterodimer, but not DNA-PK for DNA repair.
相似文献
92.
High frequency of HLA-A*0207 subtype in Chinese population 总被引:5,自引:0,他引:5
BACKGROUND: HLA-A2 is the most polymorphic and most common HLA phenotype found in various ethnic populations. Seventeen HLA-A2 subtypes have been reported and characterized by molecular techniques. Differences among these subtypes are limited to a few amino acids. Among them, HLA-A369096420761s the predominant subtype among whites. The results of a recent study, however, suggest that the HLA-A369096420761ubtype may be present at a high frequency in the Chinese population. STUDY DESIGN AND METHODS: To determine the exact frequency of the HLA-A1996 Sep s1996 Septype in the Chinese population, genomic DNA samples obtained from 54 HLA-A2-positive Chinese in Taiwan were studied by using sequence- specific primers and polymerase chain reaction. RESULTS: HLA-A360207 was present in 56 percent of the studied subjects. The estimated gene frequency for HLA-A90207 is 17.8 percent in the Chinese population. CONCLUSION: HLA-A818-21207 is the most common HLA-A2 subtype among Chinese. The high frequency of the HLA-A*0207 allele in this population offers a unique opportunity to study the ways in which different HLA-A2 subtypes may influence the clinical outcome of allograft transplantation and the disease susceptibility of recipients. 相似文献
93.
The original activated partial thromboplastin time-based assay for activated protein C (APC)-resistant factor Va (FVa) requires carefully prepared fresh plasma and cannot be used in patients receiving warfarin or in patients with antiphospholipid antibodies. A new test is described here that circumvents these limitations and distinguishes without overlap heterozygotes for APC-resistant FVa from persons with normal FV. A diluted test plasma is incubated with an FV-deficient substrate plasma and tissue factor and then clotted with Ca2+ or Ca2+ plus APC. Test results are independent of the FV level or the dilution of the test plasma used. Of 39 controls, 37 gave normal results. Two controls (5%) gave results indicative of APC resistant FVa and on DNA analysis were found to be heterozygous for FV R506Q. Twenty of 21 randomly selected patients receiving warfarin gave normal results. In the single patient with abnormal results, heterozygous FV R506Q was confirmed by DNA analysis. Two of 15 patients with protein S deficiency and 5 of 29 patients with a lupus anticoagulant had abnormal results. APC resistance caused by FV R506Q was confirmed in the five of these seven patients available for DNA analysis. APC-resistant FVa was also detected in 10 of 21 (46%) stored plasma from unrelated patients with venous thrombosis and negative earlier evaluation for a lupus anticoagulant or a deficiency of protein C, protein S, or antithrombin, which confirms a high incidence of this defect among patients with venous thrombosis. 相似文献
94.
95.
96.
Targeted gene transfer to human hematopoietic progenitor cell lines through the c-kit receptor 总被引:1,自引:0,他引:1
Schwarzenberger P; Spence SE; Gooya JM; Michiel D; Curiel DT; Ruscetti FW; Keller JR 《Blood》1996,87(2):472-478
In this report, we describe a novel gene therapy approach for hematopoietic stem/progenitor cells using a specific receptor-mediated gene transfection procedure to target c-kit+ cell lines. The vector consists of plasmid DNA containing a luciferase reporter gene that is condensed by electrostatic forces with polylysine (PL) covalently linked to streptavidin (binds biotinylated ligand) and PL covalently linked to adenovirus (AD; to achieve endosomal lysis) with the final addition of biotinylated steel factor (SLF-biotin). Targeted transfection of growth factor-dependent hematopoietic progenitor cell lines that express c-kit showed specific luciferase gene expression over cell lines that did not express c-kit. This effect was dependent on the dose of SLF-biotin and was competed by excess SLF or with monoclonal antibodies that recognize c-kit and block the binding of SLF to its receptor. Maximum transfection efficiency (> 90%) requires a 2- hour incubation period of the vector with the cells, and maximum gene expression occurred 30 hours later. Removal of the endosomalytic agent, AD, from the vector resulted in the loss of gene expression. Vector targeting was versatile and could be changed by the addition of other biotinylated ligands. In principle, this vector should be broadly applicable to deliver genes to hematopoietic stem/progenitor cells in vitro and in vivo. 相似文献
97.
Irradiation of blood components has been used to prevent transfusion-related graft-versus-host disease (GVHD) in immunocompromised patients. This study was designed to determine the effect of irradiation on neutrophil aggregation, chemotaxis, and superoxide generation. Purified neutrophils were irradiated with a Cesium source at four doses ranging from 0 to 17,500 rads. Formyl-methionyl-leucyl-phenylalanine (FMLP) and zymosan-treated serum (ZTS) cytotaxin-induced chemotaxis and migration were determined in the agarose assay. Neutrophil aggregation to FMLP was determined by aggregometry. Superoxide generation and random migration were not affected by irradiation at doses up to 17,500 rads. When compared to nonirradiated controls, the chemotactic response to ZTS remained normal, with an insignificant decline from 174 +/- 31.0 to 150 +/- 42.3 (mean +/- SD) units. The chemotactic response to FMLP declined insignificantly, from 228 +/- 31.3 at 0 rad to 207 +/- 26.4 at 17,500 rads. The aggregation response to FMLP remained within the normal range but declined from 0.78 +/- 0.11 to 0.61 +/- 0.18. At the radiation doses currently used to reduce the risk of transfusion-related GVHD, neutrophil superoxide generation and chemotactic response remain essentially normal. 相似文献
98.
Activation of the alternative complement pathway with rabbit erythrocytes by circumvention of the regulatory action of endogenous control proteins 总被引:20,自引:10,他引:20
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Cleavage of C3 by the alternative complement pathway occurs in at least two distinct phases: continuous low grade generation of C3b by the interaction of native C3, B, D, and P, and subsequent amplified cleavage of C3 by the interaction of C3b, B, D, and P which forms the amplification convertase, P,C3b,Bb. Transition to C3b-dependent amplification is necessary to achieve substantial C3 cleavage and is normally limited by the combined action of C3b inactivator (C3bINA) and βlH. An activator of the alternative pathway, such as rabbit erythrocytes (E(r)), provides sites that protect bound C3b and P,C3b,Bb from the action of these regulatory proteins and permits C3b deposited by the low grade fluid phase reaction to assemble a membrane-associated amplification convertase which can deposit additional protected C3b. Under conditions in which the control proteins, C3bINA and β1H, almost completely inactivated C3b bound to sheep erythrocytes (E(s)), which does not activate the alternative pathway, the function of C3b bound to E(r) was diminished by less than one-fifth. Further, the P- stabilized amplification convertase on E(r) was 10-fold less sensitive to β1H-mediated decay-dissociation than the convertase on E(s). The addition of E(r) to a regulated mixture of purified C3, B, D, P, C3bINA, and β1H resulted in amplified inactivation of C3 and B by formation of the amplification convertase on E(r) as indicated by its lysis with subsequent exposure to C3-C9. In contrast, E(s) did not advance the low grade fluid phase inactivation of C3 and B to amplified inactivation and the cell was not converted to an intermediate susceptible to lysis by C3- C9. Since E(r) and E(s) did not differ in their inefficient fixation of C3b generated during an unregulated fluid phase reaction, the activating capacity of E(r) must reside in its protection of bound C3b and P, C3b,Bb from the regulatory proteins rather than in enhanced capacity to bind C3b from the fluid phase. When the reaction is limited to low grade fluid phase turnover, introduction of E(r) but not E(s) results in a 100-fold increase in the deposition of C3b, indicating that surface-dependent activation of the alternative pathway is characterized by efficient deposition of C3b on the initiating surface. Thus, the activating surfaces advance the interaction of the alternative pathway proteins to the amplification phase because of the selective inability of the regulatory proteins to deal with their substrates when deposited on these surfaces and results in a specificity that is not necessarily dependent on adaptive immunity. 相似文献
99.
目的:关节软骨的修复一直是矫形外科临床治疗中的重要课题本组拟验证应用关节镜进行自体骨软骨移植治疗股骨髁关节软骨缺损的可行性。
方法:选择2001—04/2006—04淄博市中心医院骨科收治的关节镜下自体骨软骨移植治疗股骨髁关节软骨缺损患者16例,对治疗方案均知情同意。关节镜下在其非负重区的软骨面上用专用器械凿取圆柱状骨软骨,移植至软骨缺损部位以修复缺损。术后进行系统功能锻炼,随访行MRI检查及Brittberg—Peterson评分.评分标准:0分为无症状,130分表明治疗效果最差。
结果:①随访6~56个月,所有患者关节症状消失,关节活动度正常,MRI显示原关节软骨缺损区表面平整,移植骨软骨位置良好。②
术后Brittberg—Peterson评分:13例为0分,2例为2分,1例为1分。
结论:关节镜下自体镶嵌式骨软骨移植术创伤小,操作简单,能保持关节面曲度,可用于修复膝关节软骨缺损。 相似文献
100.
The underlying DNA changes associated with glucose-6-phosphate dehydrogenase (G6PD)-deficient Asians have not been extensively investigated. To fill this gap, we sequenced the G6PD gene of 43 G6PD- deficient Chinese whose G6PD was well characterized biochemically. DNA samples were obtained from peripheral blood of these individuals for sequencing using a direct polymerase chain reaction (PCR) sequencing procedure. From these 43 samples, we have identified five different types of nucleotide substitutions in the G6PD gene: at cDNA 1388 from G to A (Arg to His); at cDNA 1376 from G to T (Arg to Leu); at cDNA 1024 from C to T (Leu to Phe); at cDNA 392 from G to T (Gly to Val); at cDNA 95 from A to G (His to Arg). These five nucleotide substitutions account for over 83% of our 43 G6PD-deficient samples and these substitutions have not been reported in non-Asians. The substitutions found at cDNA 392 and cDNA 1024 are new findings. The substitutions at cDNA 1376 and 1388 account for over 50% of the 43 samples examined indicating a high prevalence of these two alleles among G6PD-deficient Chinese. Our findings add support to the notion that diverse point mutations may account largely for much of the phenotypic heterogeneity of G6PD deficiency. 相似文献