电泳迁移率法检测中药复方对白血病CD34~+CD38~-细胞核因子-κB的影响

目的 观察中药复方对急性髓系白血病(AML)CD34+CD38-干细胞群核因子(NF)-κB的影响。方法 将符合纳入标准的30例AML患者骨髓提取单个核细胞,并进行免疫磁珠分选分离人白血病CD34+CD38-干细胞群,每一例标本分为中药原方组、扶正组、祛邪组、对照组。采用电泳迁移率法(EMSA)检测中药复方对NF-κB活化的影响。结果 NF-κB活化比较分析:中药原方组抑制NF-κB活化最明显,中药原方组与扶正组、祛邪组比较差异有统计学意义(P<0.05),扶正组与祛邪组比较差异无统计学意义(P>0.05),对照组NF-κB活化最明显。结论 中药复方可以抑制NF-κB活化,在促进细胞凋亡的层次上,此实验为中医药靶向治疗AML提供了平台。     

氨磷汀对噻唑蓝实验结果的影响

［摘要］目的研究氨磷汀对噻唑蓝（MTT）实验结果的影响。方法实验共分4组：调零组（培养基）、对照组（培养基和细胞）、氨磷汀组（培养基和氨磷汀）、氨磷汀＋细胞组（培养基、细胞和氨磷汀）,如常规实验加入MTT,立即观察颜色改变及酶联免疫检测仪490 nm处测吸光度（A）值,孵育4 h后再次检测各组A值。结果①调零组、对照组加MTT后均无颜色改变,A值一直在很低水平（0.087±0.021和0.093±0.018）,孵育4 h仍无呈色反应,A值也无明显变化（0.076±0.015和0.103±0.022, P＞0.05）;②氨磷汀组、氨磷汀＋细胞组加MTT后均立即呈现蓝紫色,0.5 mmol &#8226;L 1 氨磷汀时A值分别为0.215±0.013和0.274±0.013,较调零组、对照组均明显升高（P＜0.01）,且随着氨磷汀浓度增高A值也明显升高（P＜0.01）;孵育4 h后氨磷汀组A值无继续升高（0.247±0.031, P＞0.05）,而氨磷汀＋细胞组A值仍有升高（0.381±0.022, P＜0.05）。结论氨磷汀本身及代谢产物可能与MTT直接反应并呈色,改变MTT实验中的A值,干扰MTT实验结果。     

优化噻唑蓝法检测抗肿瘤药物敏感性

［摘要］目的提高噻唑蓝（MTT）法检测抗肿瘤药物敏感性的成功率和可操作性,帮助临床合理选择抗肿瘤药。方法结合以往实践经验,在资料收集、实验前准备和实际操作等方面进行方法优化。结果优化的MTT法可操作性强,实验成本低、成功率高。结论经济有效的检测减少了不必要的医疗费用,提高了临床疗效。     

噻唑蓝比色法用于乳腺癌体外化疗药物敏感性的检测

建立肿瘤体外化疗药物敏感试验来选取有效的化疗药物,一直是临床化疗中倍受关注的问题。噻唑蓝(MTT)比色法快速、简捷,已被广泛应用于临床。本文应用MTT法检测10例乳腺癌体外化疗药物的敏感性,为临床化疗药物选择提供参考。1材料与方法1.1临床资料本组10例,年龄35~60岁。均经术中冰冻证实为乳腺恶性肿瘤,其中导管浸润癌5例,导管原位癌1例,未分类4例。1.2标本处理无菌取得手术标本,剪去脂肪、坏死组织,经含庆大霉素的生理盐水和含青霉素、链霉素的RPMI-1640培养液反复冲洗后,在200目网筛上反复剪碎组织,并用注射器针芯研磨,制成单细胞悬…     

反复呼吸道感染患儿外周血CD4~+CD25~+CD127~-调节性T细胞的表达

<正>反复呼吸道感染(RRTI)是小儿时期的常见病,其发生的原因与诸多因素有关。CD4+CD25+T调节细胞(regulatory T cell,Tr/Treg)是一个具有独特免疫调节功能的T细胞亚群。此亚群细胞可抑制性调节CD4+或CD8+T细胞活化与增殖,在免疫应答中发挥调节作用[1]。目前对CD4+CD25+CD127-调节性T细胞(Treg细胞)与RRTI的相关研究报道较少。本研     

Smoothened在急性白血病 骨髓增殖性肿瘤CD34~+细胞的表达及意义

目的探讨Hedgehog信号通路的受体Smoothened(Smo)在急性白血病、骨髓增殖性肿瘤CD34+细胞的表达及临床意义。方法收集2010年9月至2011年3月在华西医院就诊的急性白血病骨髓标本23份、骨髓增殖性肿瘤7份,并纳入10例非血液系统肿瘤作为对照,采用间接免疫荧光标记流式细胞术检测标本CD34+细胞Smo的表达量,分析比较各组Smo蛋白的表达水平。结果比较血液系统肿瘤(急性白血病、骨髓增殖性肿瘤)患者与对照组(非血液系统肿瘤)Smo表达水平,差异无统计学意义(P=0.719)。比较急性白血病、骨髓增殖性肿瘤及非血液系统肿瘤患者Smo表达水平,3组比较差异无统计学意义(P=0.934)。比较急性淋巴细胞白血病、急性髓系白血病与非血液系统肿瘤患者,3组比较差异无统计学意义(P=0.933)。结论流式细胞术检测血液系统肿瘤CD34+细胞Smo蛋白表达有临床应用价值;Smo蛋白在急性白血病、骨髓增殖性肿瘤均有表达,可作为新的治疗靶点。     

TF3-H4对CD4~+CD25~+调节性T细胞和CD4~+CD25~-效应性T细胞早期活化的影响

目的观察1',2',3',7'-四氢茶黄素-3,3'-双没食子酸酯(TF3-H4)对CD4+CD25+调节性T细胞和CD4+CD25-效应性T细胞早期活化的影响,分析TF3-H4对两群功能相反的CD4+T细胞的作用。方法免疫磁珠分离BALB/c小鼠脾脏CD4+CD25+调节性T细胞和CD4+CD25-效应性T细胞,加入ConA或PDB,和TF3-H4共同孵育12 h后收集细胞,流式细胞仪分析细胞表面早期活化标志CD69的表达。结果在ConA和PDB的作用下,两群细胞早期活化标志CD69的表达均升高。TF3-H4(20 mg.L-1)不能抑制PKC激动剂PDB激活的CD4+CD25-效应性T细胞CD69的表达,却能抑制ConA激活的CD4+CD25-效应性T细胞CD69表达;同时,TF3-H4也能明显抑制ConA激活的CD4+CD25+调节性T细胞的CD69表达。结论茶黄素衍生物TF3-H4可能经由TCR活化途径的上游抑制CD4+CD25-效应性T细胞活化;TF3-H4对CD4+CD25+调节性T细胞和CD4+CD25-效应性T细胞早期活化的抑制作用,可能是该类化合物发挥抗炎、抗肿瘤作用的机制之一。     

米托蒽醌为主的化疗方案对CD34+高表达白血病的疗效观察

目的进一步探讨米托蒽醌(MTZ)在急性白血病(AL)化疗中的作用特点,提高临床对难治性白血病的化疗疗效和AL的完全缓解(CR)率和无病生存率(FDS)。方法57例CD34 抗原高表达的AL患,随机选择3种常规化疗方案(MA／MAE、DA／DAE、HA／HAE),联合化疗1～2个疗程后分别比较缓解率、骨髓抑制及其它毒副作用;同时骨髓白血病细胞体外培养72h,进行药物杀伤效应实验,分别比较MTZ、柔红霉素(DNR)、高三尖杉酯碱(HHT)在体外对白血病细胞不同分化阶段的抑制作用。结果以MTZ为主的MA／MAE方案有效率最高,急性髓系白血病(AML)为80．0％(24／30),比DA／DAE和、HA／HAE方案高28．8％和56．9％;急性淋巴细胞白血病(ALL)的COMP方案有效率为92．6％(25／27),比CDOP方案高27．4％。药物抑制试验显示MTZ对分化较早期阶段的CD34^ 白血病细胞的抑制显高于DNR和HHT。结论MTZ具有较强的抗白血病活性,临床骨髓抑制明显和较少产生耐药,可能与其主要作用于白血病细胞的分化较早阶段有关。     

生物敷料治疗对中度烧伤患者CD4~+及CD8~+淋巴细胞影响的研究

目的:探讨中度烧伤患者早期生物敷料治疗对患者CD4+,CD8+淋巴细胞的影响。方法:将中度烧伤患者30例随机分为对照组和治疗组,治疗组烧伤后3 d内使用生物辅料治疗,对照组采用外用磺胺嘧啶银包扎治疗,在烧伤后1 d,3 d,7 d,14 d采集外周静脉血,检测各组患者淋巴细胞数目,流式细胞仪检测CD4+、CD8+T细胞的数量及其凋亡率,获得CD4+/CD8+值。观察两组临床愈合时间。结果:烧伤后1 d,3 d两组之间淋巴细胞数目、CD4+和CD8+的数量及其凋亡率、CD4+/CD8+值比较,差异无统计学意义(P>0.05),烧伤后7 d,14 d治疗组淋巴细胞数目、CD4+,CD4+/CD8+均高于对照组(P<0.05),CD4+/CD8+细胞凋亡率明显下降(P<0.05)。两组临床愈合时间实验组较对照组短,但差异无统计学意义(P>0.05)。结论:早期生物敷料治疗可以明显提高中度烧伤患者CD4+,CD8+的数量,提示生物敷料治疗可以通过免疫调理,减少创面感染机会,促进创面愈合。     

CD_7~+的急性髓细胞性白血病的临床意义

目的 :观察和探讨 CD7+ 的急性髓细胞性白血病 (AML )表达特征及它们对 AML患者预后的影响。方法 :采用间接免疫荧光法和流式细胞仪对 2 75例初诊的 AML患者进行了免疫表型检测 ,并对淋系分化抗原阳性表达率加髓系分化抗原阳性表达率大于 10 0 %患者的白血病细胞用流式细胞仪做双标记分析。结果 :2 75例 AML患者中 ,3 6例表达淋系分化抗原 CD7(占 13 .1% ) ;诱导化疗疗效方面 ,在同等化疗强度的基础上 ,CD34+ CD7+ AML患者的缓解率明显低于 CD34- CD7- AML 患者 (P<0 .0 5 )。结论 :CD7+ AML 的表达具有复杂性、异质性 ,属于一种预后不良的临床亚型 ,应进一步探索针对性的治疗策略     

Inorganic arsenic induces necrosis of human CD34-positive haematopoietic stem cells

Inorganic arsenic is a major environmental contaminant known to exert immunosuppressive effects. In this study, we report toxicity of As2O3, a trivalent inorganic form, toward isolated human hematopoietic CD34+ progenitor cells. Our results demonstrate that low concentrations of As2O3 (0.1-5 microM) inhibit in vitro proliferation of CD34+ cells and their differentiation into various hematological cell lineages. These effects were associated with the induction of a necrotic process independent of caspases and likely related to mitochondrial damage. We conclude that As2O3 can impair in vitro human hematopoiesis by decreasing survival of CD34+ progenitor cells.     

Selective toxicity of dihydroartemisinin on human CD34+ erythroid cell differentiation

Artemisinins are safely used in the combination therapy for uncomplicated malaria, but their employment during pregnancy is still controversial. In fact, animal studies reported that the active metabolite, dihydroartemisinin (DHA), causes embryonic erythrocytes depletion, when the treatment is performed during a critical period of time. The present study investigates the effect of DHA on human developmental erythropoiesis in order to characterize the target erythroid stage and to predict the window of susceptibility in human pregnancy. As a model for human developmental erythropoiesis, peripheral blood purified, CD34+ cells were committed towards erythrocytes and DHA (0.5 or 2 μM) was added to different erythroid stages during 14 days culture. Erythroid differentiation was investigated by cytofluorimetric analysis of Glycophorin A expression, by morphological analysis and erythroid globin gene expression analysis with real-time PCR. It was found that the effect of DHA was dependent on the maturation stage of erythroid cells. In fact when DHA was added to the pro- and basophilic erythroblasts caused a significant dose-dependent inhibition of cell proliferation and a significant delay of erythroid differentiation, as measured by morphological analysis, expression of Glycophorin A by immunofluorescence and of erythroid globin genes by real-time PCR. In contrast, the inhibition of stem cells and of early progenitors was transient and masked by the subsequent exponential cell growth. No effect was observed on mature erythroid stages. This is the first demonstration that DHA affects human erythropoiesis in vitro, in a dose- and time-dependent manner; the target population seems to be the pro-erythroblast and basophilic erythroblast stage, suggesting that DHA toxicity is limited to primitive human erythropoiesis. These findings outline the relevance of DHA dosage and timing to prevent embryotoxicity and support current WHO recommendations of avoiding malaria treatment with artemisinins during the first trimester of pregnancy.     

Evaluation of the antiretroviral effects of a PEG-conjugated peptide derived from human CD38

Objective: Cell infection by HIV-1 is inhibited by both the expression of CD38 and a soluble peptide (sCD38p) corresponding to its extracellular membrane-proximal amino acid sequence (amino acids 51 – 74). We show here the effects of PEG conjugation to sCD38p and provide new insights into the mechanisms behind the anti-HIV-1 effects of CD38 and derived peptides. Research design/methods: In-vitro and in-silico study. Results: PEGylation of sCD38p increased its ability to inhibit replication of HIV-1 in MT-4 cells and syncytia formation in cocultures of MT-2 and persistently HIV-1_IIIB-infected H9_IIIB cells. In silico modeling suggests that sCD38p and CD4 form stable heterodimers involving, among others, an interaction between lysine 57 (K57) of CD38 and a groove in the CD4 receptor, which, in CD4/gp120 complexes, is partially occupied by a lysine residue of the HIV-1 envelope glycoprotein. K57 substitution with a glycine in sCD38p impaired its ability to inhibit syncytia formation in MT-2/H9_IIIB cell cocultures and gp120 binding to CD4 in a mouse T cell line expressing human but not mouse CD4. Conclusions: PEGylation significantly improves the anti-HIV-1 activity of sCD38p, whose effect is probably due to competition with gp120 for a common binding site on CD4 although other mechanisms cannot be excluded so far. The inhibitory concentrations of the sCD38p-PEG as well as its poor toxicity, merit further consideration in anti-HIV-1 strategies.     

MTT法研究化疗药物对人胃癌细胞株的药效动力学特征

目的：研究化疗药物氟尿嘧啶（５－ＦＵ）、丝裂霉素ＭＭＣ（、阿霉素（ＤＯＸ）、顺铂（ＣＩＳ）对人胃癌细胞株（ＳＧＣ）的作用特征,探讨ＳＴＣ细胞株对单一及化疗药物的敏感性,化疗药物浓度及作用时间对细胞抑制率的影响。方法：ＸＧＣ朱在含有不同药物的不同浓度的培养基中孵育不同时间,用ＭＴＴ法测定细胞的抑制率,化疗药物浓度参照其在人体的最高峰值的浓度,选择其峰值浓度的１／２５－２倍范围,作用时间选择２～２４小     

神经干细胞分化调节机制及中药对其的影响

神经干细胞(NSC)是新世纪神经科学研究的热点之一,其分化调控分子研究是NSC研究的热点和难点.本文综述了NSCs增殖分化过程是细胞因子、神经递质和激素、化学物质等微环境及自身基因信号共同调节,相互作用的过程;并且对中药在NSCs分化调控的研究进展做一概述.     

中药抑抗汤含药血清对子宫内膜容受性的影响

目的探讨中药抑抗汤含药血清对体外培养的小鼠子宫内膜容受性的影响。方法40只实验小鼠随机分为两组,分别用中药抑抗汤0.8ml和等量生理盐水灌胃28d,制备含药血清和空白血清。提取妊娠第4天(D4)雌鼠子宫内膜细胞后分为含药血清(A组)、胎牛血清(B组)和空白血清(C组)三组进行细胞培养。黏附实验检测子宫内膜细胞的黏附能力,ELISA法检测细胞培养液白血病抑制因子(LIF)水平,Western blot检测整合素ανβ3蛋白水平。结果 A组细胞黏附能力、LIF和整合素ανβ3蛋白表达均较B、C组显著增加(P<0.05)。结论抑抗汤可能是通过增强细胞黏附功能,增加LIF和整合素ανβ3的表达来改善子宫内膜容受性。     

半相合移植移植物中CD34+细胞数量与临床结果相关性

目的 探讨半相合移植移植物中CD34 细胞数量对临床结果的影响.方法 对26例半相合移植患者移植物中CD34 细胞、T、B淋巴细胞数量与临床造血重建时间、急慢性移植物抗宿主病(GVHD)及存活率进行相关性分析.结果 所有患者输入CD34 细胞是(3.66～9.29)×106/kg,其中细胞数量高组(＞5.60×106/kg,A组)和低组(＜5.60×106/kg,B组)患者移植物中T、B淋巴细胞数量均无显著性差异;两组中性粒细胞造血重建时间均为11d,血小板重建时间均为18d;急性和慢性GVHD发生率A组分别为46.2%和38.5%,B组分别为46.2%和46.2%.患者总生存率A组为61.5%,B组为69.2%.结论 半相合干细胞移植输入(3.66～9.29)×106/kg的CD34 细胞是安全的,不增加GVHD的发生率,也不降低生存率.     

Comparative toxicity of known and putative metabolites of 1, 3-butadiene in human CD34(+) bone marrow cells

Species-specific susceptibility to the hematotoxic effects of 1, 3-butadiene (BD) is well known. Previous studies have revealed that murine bone marrow is uniquely susceptible to toxicity following exposure to the parent compound in vivo or exposure of bone marrow cells to the monoepoxide metabolite, 3,4-epoxybutane, in vitro. Studies described herein compare the relative ability of putative and known BD metabolites to produce concentration dependent suppression of colony formation and cytotoxicity in human CD34(+) bone marrow cells. Compounds evaluated included 3,4-epoxybutane, D, L-butane-bis-oxide, meso-butane-bis-oxide and (2S, 3R)-3-epoxybutane-1,2-diol. In contrast to results previously observed in mice, only the bis-oxides produced significant suppression of colony formation at potentially relevant concentrations (10(-8) to 10(-3) M). No enantiospecific differences were observed between the meso- and D,L-bis-oxides and no significant lineage-specific differences in susceptibility to inhibition of clonogenic response were observed among early multi-potential myeloid and erythroid hematopoietic progenitor cells. The relative potencies of the bis-oxides were found to be comparable to that of the prototype hematotoxic compound, hydroquinone. These results confirm previous studies that reveal marked species-specific differences in the susceptibility of bone marrow cells to 3,4-epoxybutane. Moreover, these results suggest that the bis-oxides of BD are capable of suppressing the clonogenic function of human hematopoietic progenitor cells, if, in fact, they are produced in human bone marrow in significant concentration. Further interpretation of these findings requires a better understanding of the metabolism of BD in humans.     

不同时期移植人脐血 CD34+细胞对大鼠脊髓损伤修复的对比研究

目的 研究不同时期移植人脐血 CD34⁺细胞修复大鼠脊髓损伤的效果和机制。方法 免疫磁珠法从人新鲜脐血中分离得到 CD34⁺细胞。雌性 Wistar 大鼠 96 只, 以 IMPACTOR MODEL-Ⅱ脊髓损伤打击器建立 T10 脊髓损伤模型, 随机均分为免疫抑制剂应用组、 损伤后急性期移植组和损伤后亚急性期移植组, 对各组后肢功能恢复情况进行 BBB 评分, 损伤中心行双重免疫荧光染色、 2,3,5-氯化三苯基四氮唑(TTC)染色和血管明胶墨汁灌注观察。结果 损伤后第 8～56 天, 细胞急性期移植组的 BBB 评分高于其余 2 组(P < 0.05); TTC 染色示组织活力降低区域比例小于其余 2 组(P < 0.01); 明胶墨汁灌注示脊髓损伤中心血管密度大于其余 2 组(P < 0.01); 亚急性期移植组的细胞存活密度大于急性期移植组（个/视野： 7.51±1.00 vs 5.51±0.89, t=6.051, P < 0.01）, 2 组均未观察到移植细胞的神经分化。结论 人脐血 CD34⁺细胞急性期移植可通过提高脊髓损伤中心血管密度促进微循环恢复, 增加组织活力,促进大鼠脊髓损伤后肢体功能恢复。