白血病患者血清IL-8的测定

目的：探讨血清白细胞介素８（ＩＬ－８）变化在白血病中的意义。方法：采用平似酶联免疫吸附法测定了１０例急性淋巴细胞白血病（ＡＬＬ）,２５例急性非淋巴细胞白血病（ＡＮＬＬ）,１１例慢性粒细胞白血病（ＣＭＬ）慢性期血清ＩＬ－８浓度。结果：ＡＬＬ、ＡＮＬＬ和ＣＭＬ患者血清ＩＬ－８水平显著高于正常对照组,ＡＬＬ与ＡＮＬＬ患者无显著性差异,急性单细胞白血病组水平显著高于急粒白细胞病（Ｍ１￣Ｍ３）组,治疗有铲的     

用流式细胞仪测定小儿急性白血病细胞DNA及其临床意义研究

应用流式细胞仪（ＦＣＭ）对９４例急性白血病（ＡＬ）骨髓单个核细胞（ＭＮＣ）进行了ＤＮＡ含量测定。在４７例初诊／复发ＡＬ中，５０％（１７／３４）的急性淋巴细胞白血病（ＡＬＬ）及３８．５％（５／１３）的急性非淋巴细胞白血病（ＡＶＬＬ）检出ＤＮＡ异倍体；４７例ＡＬ完全缓解（ＣＲ）期患儿异倍体检出率为１０．６％。１４例非白血病患儿无异倍体检出。ＡＬ初诊病例的Ｃ＿０／Ｇ＿１％高于ＣＲ及对照组，Ｓ％及（Ｓ＋Ｇ＿２）／Ｍ％低于ＣＲ及对照组；复发病例（Ｓ＋Ｇ＿２）／Ｍ％高于初诊组；在ＡＬ－ＣＲ组，Ｓ％和（Ｓ＋Ｇ＿２）／Ｍ％均与对照组无显著性差异。２０例ＡＬ的自身对照发现，１１例初诊、复发时的异倍体在ＣＲ期消失。     

205例儿童急性淋巴细胞白血病疗效分析

报告２０５例小儿急性淋巴细胞白血病（ＡＬＬ），诱导缓解期采用强烈ＣＯＤＰ＋Ｌ－ＡＳＰ和ＣＯＡＰ＋Ｌ－ＡＳＰ方案，联合大剂量氨甲喋呤（ＨＤ－ＭＴＸ）及头颅放疗（１８Ｇｙ）进行庇护所预防。使５年无病生存率（ＥＦＳ）达到７４．４％，中枢神经系统白血病复发率降至１．５％，睾丸白血病复发率降至３．４％。治疗效果表明：早期强烈化疗及有效的庇护所预防是减少骨髓及髓外复发，提高ＥＦＳ的关键。     

白血病患者纤溶活性的测定

目的：为了解白血病患者血浆纤维蛋白原（ＦＩＢ）、纤溶酶原（ＰＬＧ）和Ｄ－二聚体（Ｄ－ｄｉｍｅｒ）的活性变化。方法以ＳＴＡ Ｃｏｍｐａｃｔ全自动血凝仪及ＡＣＬ－２００型自动血凝仪测定２０例正常健康人、１２例急性原始粒细胞白血病（Ｍ１、Ｍ２）、１０例急性早幼粒细胞白血病（Ｍ３）、８例急性淋巴细胞白血病（ＡＬＬ）、６例慢性粒细胞白血病（ＣＭＬ）患者血浆的纤维蛋白原含量（ＦＩＢ）、纤溶酶原活性（ＰＬＧ）及     

急性髓系白血病细胞外基质纤连蛋白、基膜连接蛋白的分析及临床意义

目的探讨细胞外基质纤连蛋白（Ｆｎ）、基膜连接蛋白（Ｌｍ）与白血病细胞生长、分化之间的关系。方法应用免疫荧光法观察１８例急性髓细胞白血病（ＡＭＬ）骨髓细胞长期培养（ＬＴＢＭＣ）细胞外基质Ｆｎ、Ｌｍ含量的变化。结果①９例获独立生长的ＡＭＬ（ＡＭＬｓｍ）骨髓细胞在培养的１～３周,Ｆｎ、Ｌｍ含量均显著低于９例未获独立生长的ＡＭＬ（ＡＭＬｎｓｍ）;②在培养的第３周,８例ＡＭＬｎｓｍ幼稚细胞比例可降至０．０５以下,而７例ＡＭＬｓｍ未能降至０．０５;③各周幼稚细胞比例分别与Ｆｎ、Ｌｍ含量相关分析示：第０～８周正常对照均呈正相关,０～４周ＡＭＬｎｓｍ均呈负相关而ＡＭＬｓｍ仅Ｆｎ组呈负相关。结论Ｆｎ和（或）Ｌｍ的改变在ＡＭＬ发生、发展的某一环节起重要作用。此外,Ｆｎ、Ｌｍ含量在ＡＭＬｓｍ与ＡＭＬｎｓｍ之间不同,这对ＬＴＢＭＣ应用于ＡＭＬ自身骨髓体外净化病例的选择可能有参考价值。     

白细胞介素8及其受体在急性白血病的表达及其意义

目的探讨白细胞介素８（ＩＬ８）及其Ａ、Ｂ受体（ＩＬ８ＲＡ、ＩＬ８ＲＢ）的表达与急性白血病（ＡＬ）的分型、疗效及并发症的关系。方法用酶联免疫夹心法检测７７例初诊ＡＬ患者外周血ＩＬ８含量,用间接免疫荧光及流式细胞术检测骨髓单个核细胞（ＭＮＣｓ）表达ＩＬ８Ｒ,并检测了１５例ＡＬ完全缓解（ＣＲ）期脑脊液（ＣＳＦ）中ＩＬ８水平。结果ＡＬ患者外周血ＩＬ８水平高于正常人,其中急性髓系白血病（ＡＭＬ）高于急性淋巴细胞白血病（ＡＬＬ）,Ｍ４、Ｍ５高于Ｍ１～Ｍ３,ＢＡＬＬ高于ＴＡＬＬ（Ｐ＜０．０５）。ＡＬＬ中ＩＬ８＞１００ｎｇ／Ｌ组比≤１００ｎｇ／Ｌ组疗效差（Ｐ＜０．０５）;约３６．３６％的ＡＬ患者ＩＬ８Ｒ（＋）,ＩＬ８Ｒ（＋）者的外周血白细胞计数及幼稚细胞比例显著高于ＩＬ８Ｒ（－）者（Ｐ＜０．０５）;ＣＲ期患者ＣＳＦ中ＩＬ８水平与治疗前比较,差异无显著性（Ｐ＞０．０５）,合并中枢神经系统白血病（ＣＮＳＬ）时明显升高。结论外周血和ＣＳＦ中ＩＬ８水平的检测及检测骨髓ＭＮＣｓ膜ＩＬ８Ｒ的表达有助于ＡＬ各亚型的鉴别及预后判断;ＣＮＳＬ发生时ＣＳＦ中ＩＬ８增高。     

用PCR技术评价微量残留白血病细胞体外净化的效果

应用系列稀释法ＰＣＲ技术检测含５％携带ＰＣＲγ基因重排的ＣＣＲＦ－ＣＥＭ株细胞的模拟缓解期白血病骨髓的微量残留白血病细胞，敏感性为１０＾－＾５－１０＾－＾６水平。经ＶＰＬ加ＡＤＭ，Ｖｐ１６，ＶＣＲ联合体外净化１小时，结果显示有３个对数级的杀伤作用。用体外克隆形成培养法观察该联合方案对急性期白血病患者白病祖细胞集落，（ＣＦＵ－Ｌ）及正常骨髓造血干细胞集落（ＣＦＵ－ＧＭ）的抑制作用，ＣＦＵ－ＧＭ存活率     

中期荧光原位杂交在急性早幼粒细胞白血病诊断和微小残？…

为研究中期荧光原位杂交（Ｍ－ＦＩＳＨ）在急性早幼粒细胞白血病（ＡＰＬ）诊断及其微小残留病（ＭＲＤ）检测中的价值，应用１７号全染色体彩涂探针对１０例初诊时按细胞形态学诊断为急性髓细胞白血病（ＡＭＬ）的未治疗患（初治ＡＰＬ５例，复发ＡＰＬ３例，ＡＭＬ－Ｍ１和ＡＭＬ－Ｍ５各１例）和１０例治疗后获完全缓解（ＣＲ）的ＡＰＬ患作耻Ｍ－ＦＩＳＨ检测，并与常规细胞遗传学和逆转录－聚合酶链反应（ＲＴ－ＰＣＲ）结     

不同类型急性白血病用MAC方案预处理自体骨髓移植疗效的比较研究

为探讨马法兰、环磷酰胺和阿糖胞苷（ＭＡＣ）预处理自体骨髓移植（ＡＢＭＴ）治疗不同类型白血病的疗效，比较了１５例急性髓细胞白血病（ＡＭＬ）和９例急性淋巴细胞白血病（ＡＬＬ）用ＭＡＣ方案预处理ＡＢＭＴ的疗效。结果：ＭＡＣ方案对两类急性白血病患者均有明显的造血抑制及抗白血病作用。ＭＡＣ方案作预处理ＡＢＭＴ后ＡＭＬ患者比ＡＬＬ患者复发率低，生存期长［２４（３～６２）个月ｖｓ１４（１～２６）个月］。提示：ＭＡＣ预处理方案适用于ＡＭＬ患者，而对ＡＬＬ患者作ＡＢＭＴ需调整或改换其它方案。     

执业医师、医师晋升、研究生入学考试辅导试题撷英(六)——小儿血液病

题目Ａ型题（第１题至第２０题）请从Ａ、Ｂ、Ｃ、Ｄ、Ｅ１个最佳答案。１．关于小儿白血病，下列哪项是错误的？Ａ．占儿童恶Ｂ．急性淋巴细胞白血病（ＡＬＬ）是小儿白Ｃ．小儿ＡＬＬ的预后较成人好；Ｄ．骨髓Ｅ．为预防中枢神经病（ＣＮＳＬ），所有小儿白血病均应进行头颅放射疗（放疗）。２．关于小儿急性粒细胞白血病（ＡＭＬ），下列哪项是Ａ．Ａｕｅｒ小体阳性是预后不良的因素；Ｂ．ＤＡ方案Ｃ．Ｄ．继发于骨髓增生异常征（ＭＤＳ）的ＡＭＬ预后不良；Ｅ．头颅放疗不是预防的常规手段。３．关于ＭＤＳ，下列哪项是正确的？Ａ．干扰…     

白血病患者血管新生及相关因素研究

目的　观察白血病患者骨髓血管新生情况,分析血管新生相关因素内皮抑素(ES)、血管内皮生长因子(VEGF)及其受体(VEGFR)在急、慢性白血病患者中的表达及意义。方法　应用血管性血友病因子(vWF)抗体标记免疫组化法观察骨髓血管新生情况,ELISA法测定血浆ES和VEGF水平,流式细胞术(FCM)分析新鲜分离的白血病细胞表面VEGFR表达。结果　26例初诊的急性白血病(AL)和 5例慢性粒细胞白血病(CML)患者骨髓微血管密度 (MVD)分别为 (20. 78±7. 75) /高倍镜视野(HP)和(28. 67±7. 32) /HP,较对照组 [ (9. 29±3. 53) /HP]明显增高 (P<0. 01),治疗后获缓解者骨髓MVD[AL缓解患者为(11. 33±5. 66) /HP,CML缓解患者为(17. 00±8. 04) /HP]较治疗前明显降低(P<0. 05和<0. 01),AL缓解组与对照组比较差异无统计学意义(P>0. 05)。AL和CML患者血浆ES、VEGF明显高于对照组(P值均<0. 05),化疗后获完全缓解(CR)的AL患者血浆ES和VEGF含量和CML CR患者的ES水平均降低至正常水平 (与对照组相比,P值均 >0. 05),但CML患者的VEGF水平仍高于正常对照(P<0. 01)。AL患者骨髓单个核细胞表面VEGFR均有不同程度的高表达,而 3例CML和 5例正常对照骨髓均为VEGFR低表达。结论　初诊白血病患者存在骨髓血管新生及血浆ES、VEGF水平增高,AL细胞不同程     

Bone marrow extracellular matrix molecules improve gene transfer into human hematopoietic cells via retroviral vectors.

Direct contact between hematopoietic cells and viral packaging cell lines or other sources of stroma has been shown to increase the efficiency of retroviral-mediated gene transfer into these target cells compared with infection with viral supernatant. We have investigated the role of defined bone marrow extracellular matrix molecules (ECM) in this phenomenon. Here we report that infection of cells adhering to the carboxy-terminal 30/35-kD fragment of the fibronectin molecule (30/35 FN), which contains the alternatively spliced CS-1 cell adhesion domain, significantly increases gene transfer into hematopoietic cells. Two retroviral vectors differing in recombinant viral titer were used. Gene transfer into committed progenitor cells and long-term culture-initiating cells, an in vitro assay for human stem cells, was significantly increased when the cells were infected while adherent to 30/35 FN-coated plates compared with cells infected on BSA-coated control plates or plates coated with other bone marrow ECM molecules. Although gene transfer into committed progenitor cells and to a lesser degree into long-term culture-initiating cells was increased on intact fibronectin as well, increased gene transfer efficiency into hematopoietic cells on 30/35 FN was dependent on CS-1 sequence since infection on a similar FN fragment lacking CS-1 (42 FN) was suboptimal. 30/35 FN has previously been shown by our laboratory and other investigators to mediate adhesion of primitive murine and human hematopoietic stem cells to the hematopoietic microenvironment. Additional studies showed that neither soluble 30/35 FN nor nonspecific binding of hematopoietic cells to poly-L-lysine-coated plates had any appreciable effect on the infection efficiency of these cells. Our findings indicate that hematopoietic stem cell adhesion to specific ECM molecules alters retroviral infection efficiency. These findings should aid in the design of gene transfer protocols using hematopoietic progenitor and stem cells for somatic gene therapy.     

EPOR在急性白血病细胞的表达及其临床意义

本研究旨在探讨红细胞生成素受体(EPOR)在急性白血病(AL)细胞上的表达及其临床意义。收集我院门诊及住院40例急性白血病患者骨髓,并选择24例骨髓正常患者的骨髓作为对照。应用逆转录聚合酶链反应(RT-PCR)方法检测白血病细胞EPOR mRNA的表达。结果显示,急性白血病细胞有EPOR的表达,表达率为57.5%,平均表达量(灰度值)为0.3549±0.2800,均低于正常对照(p<0.05)。急性髓系白血病(AML)与急性淋巴系白血病(ALL)表达率差别无统计学意义(p>0.05),表达量灰度值AML高于ALL(p<0.05)。结论:急性白血病细胞表达EPOR,表达率和平均表达量均低于正常对照(p<0.05)。AML与ALL表达率无显著差异(p>0.05),表达量AML高于ALL(p<0.05)。     

急性白血病患者骨髓VEGF高表达及其与预后的相关性分析

目的:检测非M3型急性白血病(AL)患者的骨髓血管内皮生长因子(VEGF)水平,探讨VEGF的表达水平与预后的关系.方法:选取2016年1月至2019年12月在武威市人民医院血液内科住院的AL患者共114例,以本课题组25名健康志愿者为对照组,用ELISA方法分别检测骨髓VEGF的浓度,依据VEGF表达水平进行分组,比...     

Apert's syndrome: differential in vitro production of matrix macromolecules and its regulation by interleukins

During embryonic development, variations in the composition of the extracellular matrix (ECM) macromolecules influence bone tissue differentiation. We present novel findings on the in vitro phenotypic expression of periosteal fibroblasts obtained from patients affected by Apert's syndrome, a rare craniofacial malformation, and the effects that interleukins (ILs) induce on the phenotype. Apert fibroblasts synthesized greater quantities of glycosaminoglycans (GAGs) and intracellular type I collagen, and also produced more type III collagen and fibronectin. The amount of hyaluronic acid (HA) secreted by Apert fibroblasts was much higher than that secreted by normal fibroblasts, but, as the absolute values of heparan sulphate (HS), chondroitin sulphate (CS) and dermatan sulphate (DS) also rose in Apert media, the HA–sulphated GAG ratio was similar in the media obtained from both populations. Both ILs triggered elevations of HA in normal cells, although relative percentage secretion remained unaltered, but significantly reduced HA secretion by Apert cells. IL-1 significantly increased CS in normal and Apert media, whereas IL-6 enhanced HS and DS in media of both populations. HA–sulphated GAG ratio decreased in Apert media after IL treatment. Both ILs boosted fibronectin production by Apert fibroblasts, whereas IL-1 increased type III but not type I collagen. Taken together, these data demonstrate that the synthesis and secretion of ECM macromolecules are markedly altered in Apert fibroblasts. The fact that treatment with ILs further modifies the Apert phenotype suggests that ILs may be implicated in the pathophysiology of the malformations during skull morphogenesis.     

Chfr基因在急性白血病骨髓细胞中表达的研究

为了探讨急性白血病（AL）患者骨髓细胞中有丝分裂早期检查点Chfr基因的表达及其意义，对46例初诊未治疗的AL患者骨髓单个核细胞，用RT—PCR和免疫组织化学的方法检测Chfr基因mRNA及蛋白的表达，并与正常人骨髓单个核细胞进行对照。结果表明：免疫组织化学和RT—PCR分析显示。28例急性髓细胞白血病（AML）中的15例，18例急性淋巴细胞白血病（ALL）中的13例骨髓单个核细胞Chfr基因mRNA和蛋白的表达与正常对照相比显著下降，同时Chfr基因表达异常病例染色体检查存在核型异常。结论：急性白血病骨髓细胞有丝分裂检查点Chfr基因表达受损，可能在白血病发病中具有重要作用。     

细胞周期相关因子cdtl和geminin在急性白血病患者中的表达

本研究旨在检测geminin和cdtl在初诊急性白血病(AL)患者外周血或骨髓细胞中的表达水平,并进一步探讨其在AL发病机制中的作用.采用SYBR Green实时定量逆转录聚合酶链反应(SYBR-RT-PCR)技术,检测初诊AL患者外周血或骨髓细胞geminin和cdtl mRNA的表达.结果表明:13例ALL初诊患者外周血中有10例孪蛋白(geminin)和cdtl mRNA表达阳性,阳性率为76.92%;14例AML初诊患者外周血中有9例表达阳性,阳性率为64.29%;而正常对照组中无阳性表达(0/10).ALL和AML初诊患者骨髓细胞中geminin和cdtl mRNA表达(ALL:108.06±67.34,52.37±35.16;AML;62.66±58.69,26.68±22.29)均高于对照组(11.81±2.83,7.32±5.77),差异有统计学意义(p＜0.01,p＜0.05);34例初诊AL患者骨髓细胞中geminin和cdtl mRNA的表达呈明显的正相关(r=0.55,p＜0.01).结论:geminin与cdtl在初诊AL患者中表达增高,且两者在AL骨髓细胞呈正相关,其过度表达可能在AL的发病机制中起重要作用.     

CD^＋34干／祖细胞与纤维连接蛋白的粘附在慢性粒细胞白血？…

探讨ＣＤ＾＋３４干／祖细胞与纤维连接蛋白（ＦＮ）的粘附在慢性粒细胞白血病（ＣＭＬ）发病中的作用。方法①采用细胞术双标记法检测初治ＣＭＬ慢性期３０例正常骨髓１０份ＣＤ＾＋３４细胞上整合素β１链（ＣＤ２９）和α４链（ＣＤ４９ｄ）的表达;②结晶紫染色法观察免疫磁珠分选的ＣＭＬ慢性期５例和５名正常人骨髓ＣＤ＾＋３４细胞ＦＮ的粘附功能;③极限稀释液体微培养观察ＦＮ对正常和ＣＭＬ慢性期骨髓粒细胞－巨噬细胞祖细     

Functionalization of porous BCP scaffold by generating cell‐derived extracellular matrix from rat bone marrow stem cells culture for bone tissue engineering

The potential of decellularized cell‐derived extracellular matrix (ECM) deposited on biphasic calcium phosphate (BCP) scaffold for bone tissue engineering was investigated. Rat derived bone marrow mesenchymal stem cells were cultured on porous BCP scaffolds for 3 weeks and decellularized with two different methods (freeze–thaw [F/T] or sodium dodecyl sulfate [SDS]). The decellularized ECM deposited scaffolds (dECM‐BCP) were characterized through scanning electron microscopy, energy dispersive X‐ray spectrometer, and confocal microscopy. The efficiency of decellularization was evaluated by quantifying remaining DNA, sulfated glycosaminoglycans, and collagens. Results revealed that F/T method was more effective procedure for removing cellular components of cultured cells (95.21% DNA reduction) than SDS treatment (92.49%). Although significant loss of collagen was observed after decellularization with both F/T (56.68%) and SDS (70.87%) methods, F/T treated sample showed higher retaining amount of sulfated glycosaminoglycans content (75.64%) than SDS (33.28%). In addition, we investigated the cell biocompatibility and osteogenic effect of dECM‐BCP scaffolds using preosteoblasts (MC3T3‐E1). Compared to bare BCP scaffolds, dECM‐BCP_F/T scaffolds showed improved cell attachment and proliferation based on immunofluorescence staining and water soluble tetrazolium salts assay (p < .001). Moreover, dECM‐BCP scaffolds showed increased osteoblastic differentiation of newly seeded preosteoblasts by up‐regulating three types of osteoblastic genes (osteopontin, alkaline phosphatase, and bone morphogenic protein‐2). This study demonstrated that functionalization of BCP scaffold using cell‐derived ECM could be useful for improving the bioactivity of materials and providing suitable microenvironment, especially for osteogenesis. Further study is needed to determine the potential of dECM‐BCP scaffold for bone formation and regeneration in vivo.     

靶向骨髓微环境治疗急性白血病的研究进展

急性白血病(AL)是一类起源于白血病干细胞(LSC)的恶性克隆性疾病,且死亡率高.尽管联合化疗和造血干细胞移植明显改善了AL患者的完全缓解率和总体生存期,但耐药和复发仍是治疗的难题.越来越多的研究表明,白血病耐药和复发的原因之一是骨髓微环境对LSC的保护作用,LSC隐蔽在骨髓微环境中从而逃避化疗药物的杀伤,并在骨髓微环境内自我更新,还可通过改变骨髓微环境而促进AL的发展.因此,通过靶向调控骨髓微环境与LSC间的相互作用,有望找到治疗AL的新方法.