全反式维甲酸对急性早幼粒细胞白血病患者血清学的影响

目的：探讨全反式维甲酸（All trans retinoic acid,ATRA）作为辅助剂在急性早幼粒细胞白血病（Acute promyelocytic leukemia,APL）治疗中,对患者血清促红细胞生成素（Erythropoietin,EPO）、血清铁蛋白、叶酸和维生素B12水平的影响。方法：回顾性分析我院2011 年6 月-2015 年6 月接诊的50 例急性早幼粒细胞白血病患者的临床资料。根据患者的治疗方法不同将患者分为两组,A 组（亚砷酸钠治疗组）和B 组（亚砷酸钠联合ATRA）。对比两组患者治疗后血清铁蛋白、EPO、叶酸和维生素B12 恢复情况。结果：两组患者入院时所有血清EPO、铁蛋白等比较,差异无统计学意义（P>0.05）;治疗后两组患者血清各指标均有所改善（P<0.05）;治疗后B 组患者中EPO、血清B12、血清铁蛋白和叶酸恢复正常水平者明显多于A 组,差异具有统计学意义（P<0.05）。结论：全反式维甲酸辅助治疗急性早幼粒细胞白血病患者能够更好的改善患者血清EPO、铁蛋白、叶酸和维生素B12 的异常,对于疾病的治疗有一定的效果。     

急性早幼粒细胞白血病巩固治疗现状及进展

急性早幼粒细胞白血病(APL)曾被认为是最迅速的致命白血病,特点为临床表现凶险,早期死亡率高,治愈率低。药物全反式维甲酸及亚砷酸的应用,使APL的治疗取得了很大成功,其完全缓解率可达90%。然而APL的复发率仍然较高,约15%-30%。降低复发率和提高长期生存已成为研究重点,如何选择合理的缓解后治疗策略至关重要。缓解后治疗一般包括巩固治疗和维持治疗,而最佳治疗方案的确定仍然有待商榷。因此,本文就APL缓解后巩固治疗回顾相关文献进行整合分析,综述APL巩固治疗的研究进展。     

成人和儿童急性早幼粒细胞白血病患者应用三氧化二砷的对比研究

目的:探讨持续慢点三氧化二砷(As2O3)治疗儿童和成人急性早幼粒细胞白血病(APL)患者的疗效。方法:选取2007-2011年来我院就诊并采取持续慢点As2O3方式治疗的APL患者60例,其中儿童28例,成人32例。成人As2O3剂量为0.16 mg/kg,儿童为0.08 mg/kg,每分钟8滴,18-21 h完成,28天为一疗程。测量两组不同时间点的血总砷及血三价砷浓度,并记录治疗过程中高白细胞血症的发生率及高白细胞血症持续时间。结果:儿童中高白细胞血症的发生率为60.71%(17/28),明显高于成人的34.38%(11/32),差异有统计学意义(P0.05)。成人高白细胞血症平均持续时间为7.4天(6-10天),明显高于儿童的5.53天(4-8天),差异有统计学意义(P0.01)。成人不同时间点血总砷浓度高于儿童(P0.01),同一时间点血三价砷浓度高于儿童;随着用药时间延长,成人和儿童血总砷浓度和三价砷浓度增加,差异有统计学意义(均P0.01),于用药第14天达稳态。结论:儿童高白细胞血症发生率高于成人,持续时间短,血总砷及三价砷浓度水平低于成人。     

自噬与急性早幼粒细胞白血病

急性早幼粒细胞白血病(APL)是急性髓性白血病的一种亚型,其分子特征是具有t(15;17)(q22;q21)染色体易位,并形成融合肿瘤蛋白,进而阻止早幼粒细胞分化成熟。全反式维甲酸(ATRA)和三氧化二砷(ATO)作为经典的治疗APL的药物,能够通过转录调节并激活泛素-蛋白酶体通路,促进融合肿瘤蛋白降解,发挥其临床抗白血病的功效。最近的研究发现,ATRA与ATO均能够诱导APL细胞自噬,且自噬在融合肿瘤蛋白降解及诱导早幼粒细胞分化中发挥至关重要的作用。我们简要综述近年来APL的研究进展及自噬在APL治疗中的作用。     

早幼粒细胞白血病分子遗传学诊疗技术在临床中的应用

目的:综合多种急性早幼粒细胞白血病(APL)分子遗传学检测方法,比较染色体核型分析(CC)检测方法在临床诊疗中的应用优势.方法:采用急性早幼粒细胞白血病(APL)临床诊断中常用的分子生物学(反转录筑巢式聚合酶链反应--Rf-nest-PCR)、细胞形态学和荧光原位杂交(FISH)方法分别对83例来本院初、复诊的APL患者的骨髓标本进行分析,将结果分别与染色体核型分析(CC)的结果进行比较.结果:83例APL患者中染色体诊断出现典型的t(15;17)异位为79例,占总人数的95.2％;2例出现复杂染色体变化即t(15;17)+7和t(15;17)+9,占总人数的2.4％;1例出现t(11;17)异-位,占总人数的1.2％;1例为正常.PCR对融合基因检测阳性率为92.8％;细胞形态学检测结果阳性率为92.8％;荧光原位杂交阳性率为97.6％.结论:染色体核型分析是对APL疾病诊断的可靠法,特别是在对一些复杂核型的判断上相对于PCR和细胞形态学以及FISH的检测方法上均有很大优势,是其他诊断方法无法取代的.     

全反式维甲酸及依托泊苷对急性早幼粒细胞白血病细胞磷脂酰丝氨酸暴露及促凝血活性的影响

目的：探讨磷脂酰丝氨酸（PS）暴露在急性早幼粒细胞白血病（APL）细胞促凝血活性中的作用及不同药物对其产生的影响。方法：实验共分为4组：新采集APL细胞组、APL细胞单纯培养组、APL细胞全反式维甲酸（ATRA）处理组及APL细胞依托泊苷（VP16）处理组。提取10名初发APL患者的骨髓APL细胞进行实验,提取10名健康成人外周血单个核细胞作为凝血实验的正常对照。分别用1μmol·L-1ATRA和1μmol·L-1VP16处理APL细胞24 h,利用共聚焦显微镜及流式细胞术检测各组细胞PS暴露情况。利用凝血实验检测各组细胞总的促凝活性及细胞表面磷脂的促凝血活性。利用PS特异结合蛋白乳粘素对各组细胞进行凝血抑制实验。结果：新采集的APL细胞存在一定量的PS外翻,并且与外周血单个核细胞相比,存在更高的促凝血活性（P〈0.05）,ATRA对APL细胞的PS外翻及促凝活性有抑制作用（P〈0.05）,VP16则对其有显著的促进作用（P〈0.001）。乳粘素可以拮抗APL细胞至少70%的促FXa和FIIa生成活性。结论：PS暴露在APL细胞促凝血过程中发挥着重要作用。分化治疗药物ATRA和化疗药物VP16分别通过减少和增加APL细胞表面PS的暴露来减轻和加重凝血紊乱。乳粘素通过与PS特异结合可以有效地阻断暴露的PS的促凝活性,是一种潜在的治疗APL凝血紊乱的抗凝剂。     

适用于水稻叶片蛋白质组分析的双向电泳技术

针对水稻叶片中含有大量色素和酚等干扰物质的现象,通过对水稻叶片蛋白提取方法、上样量和聚丙烯酰胺凝胶浓度等方面做了必要改进,建立了一套适用于水稻叶片蛋白质组分析的双向电泳（2-DE）方法。     

持续缓慢静脉输注三氧化二砷治疗急性早幼粒细胞白血病的临床护理效应

目的:观察持续缓慢输注三氧化二砷(As_2O_3)治疗急性早幼粒细胞白血病(APL)病人的临床护理效应。方法:对196例急性早幼粒细胞白血病患者(APL)给予持续缓慢静脉输注As_2O_3治疗,每例患者As_2O_3日治疗总量按0.16mg/kg计算,As_2O_3注射液10毫克(10 mg/支),加入5%葡萄糖500毫升(或生理盐水500毫升)静脉滴注,8-10滴/分钟,每次滴注约18-21小时,连续用药28-50天,至完全缓解(CR)。同时,注重心理护理,加强输液管理,在用药过程中严密观察药物的毒副作用和对不良反应的及时对症护理等措施。结果:177例病人完全缓解(CR),完全缓解率达90.3%。结论:应用持续缓慢静脉输注As_2O_3治疗APL,实施积极有效的护理措施是取得显著疗效的重要环节。     

急性早幼粒细胞白血病患者小孢根霉变种感染1例

报道1例由小孢根霉变种导致的皮肤毛霉病.患者女,30岁.因患急性早幼粒细胞白血病,化疗后继发口腔颌面部小孢根霉变种感染,表现为发热、右侧颊黏膜水肿,皮损中央溃疡、焦痂,周边组织炎性水肿,以面颊部、颌下区为中心明显肿胀,逐渐累及右侧颈部及右下颌,进行性加重.坏死组织涂片镜检显示有粗大、无分隔直角菌丝,真菌学检查鉴定为小孢根霉,分子测序证实为小孢根霉变种.给予两性霉素B、伊曲康唑静脉滴注和手术清创,坏死组织连续3次真菌培养均未培养出小孢根霉变种,患者体温逐渐恢复正常,治疗2周后颔面部肿胀明显减退,浅表淋巴结未触及肿大,4周后额部及右眼睑肿胀已完全消退,伤口结痂愈合.随访2 a,右侧颌下可见长约2 cm手术疤痕,未见皮肤毛霉病复发.     

三氧化二砷对急性早幼粒细胞白血病细胞微粒数量及促凝活性的影响

目的:观察急性早幼粒细胞白血病(APL)细胞来源微粒(APL-MP)的促凝活性、表面组织因子(TF)表达情况、TF在其促凝活性中发挥的作用及分化治疗药物三氧化二砷(ATO)对上述指标有何影响。方法:选取3例初发APL患者,提取骨髓APL细胞,3名缺铁性贫血患者提取骨髓单个核细胞作为对照。分别用不同浓度ATO处理APL细胞24 h、48 h、72 h,收集细胞培养液提取微粒。采用流式细胞术对微粒进行定量分析并进行微粒表面TF表达情况检测;利用凝血实验比较不同组细胞释放微粒的促凝血活性;应用抗TF抗体抑制微粒促凝血活性实验检测TF在APL-MP的促凝血活性中发挥多大作用。结果:1.0μM及2.0μM ATO能显著促进APL细胞释放微粒。与正常骨髓来源单个核细胞释放的微粒相比,骨髓APL-MP的TF表达及促凝活性均显著增高,0.5μM及1.0μM ATO处理可以有效降低APL-MP的TF表达及促凝活性,且这一作用呈时间依赖性。各组APL-MP经抗TF抗体孵育后凝血时间显著延长。结论:APL-MP的TF表达和促凝学活性均显著增高,并且TF在APL-MP的促凝血活性中发挥着重要作用。ATO能显著促进APL细胞释放微粒,低浓度ATO可以有效降低APL-MP的TF表达及促凝血活性。     

抗生素AGPM发酵过程中蛋白质表达差异的研究

由土壤中筛选到一能产生新型抗肿瘤抗生素AGPM的藤黄灰链霉菌株,应用蛋白质双向电泳方法,比较了藤黄灰链霉菌在发酵24h与72h蛋白质表达的差异,发现在发酵72h有17个蛋白质差异点出现,此外还发现一些蛋白质的含量明显高于24h的同种蛋白含量,这表明这些蛋白可能与抗生素AGPM的合成以及和菌体的生长等有关。     

Proteomic signature of human embryonic stem cells

Human embryonic stem cells (hESC) represent a population of undifferentiated pluripotent cells with both self-renewal and multilineage differentiation characteristics. Proteomics provides a powerful approach for studying the characteristics of hESC and discovering molecular markers. We have analyzed proteome profiles of three hESC lines using 2-DE and MALDI TOF-TOF. Out of 844 spots analyzed with MALDI TOF-TOF, 685 proteins were identified of which 60 proteins were classified as the most abundant proteins on 2-D gels. A large number of proteins particularly high abundant ones were identified as chaperones, heat shock proteins, ubiquitin/proteasome, and oxidative stress responsive proteins underscoring the ability of these cells to resist oxidative stress and increase the life span. Several proteins involved in cell proliferation and differentiation were also among the highly expressed proteins. Although overall expression pattern of three hESC were similar, 54 spots changed quantitatively and 14 spots changed qualitatively among the hESC cell lines. Most of these proteins were identified as proteins involved in cell growth, metabolism and signal transduction, which may affect the self-renewal and pluripotency. To our knowledge, this study represents the first proteomic dataset for hESC and provides a better insight into the biology of hESC. Proteome maps of hESC are accessible at http://www.RoyanProteomics.ir.     

Improvement of two-dimensional gel electrophoresis proteome maps of the haloarchaeon Haloferax volcanii

Proteins of haloarchaea are remarkably unstable in low-ionic-strength solvents and tend to aggregate under standard two-dimensional (2-D) gel electrophoresis conditions, causing strong horizontal streaking. We have developed a new approach to generate 2-D maps of halophilic proteins which included washing cells with 1.5 M Tris-HCl buffer. In addition, proteins were precipitated with acetone, solubilized with urea and thiourea in the presence of the sulfobetaine detergent 3-[(3-cholamidopropyl)dimethylamino]-1-propanesulfonate (CHAPS), reduced with tributylphosphine (TBP), and separated with microrange strips of immobilized pH gradients (pH 3.9-5.1). This combination enabled the construction of highly reproducible 2-D maps of Haloferax volcanii proteins.     

人肺巨细胞癌蛋白质组的二维电泳和计算机图象分析

为优化用于蛋白质组研究的二维电泳技术和计算机图象分析技术 ,以及初步分析比较与肿瘤细胞转移相关的蛋白质 ,以人肺巨细胞癌 (PLA- 80 1 - D、C)高、低转移株作为研究对象 ,应用 IPG-phor进行第一向等电聚焦 ,随后 ,在 Protein IPG conversion Kit上进行垂直 SDS- PAGE的分离 .利用光密度仪对银染的凝胶扫描 ,通过 PDQuest软件进行蛋白斑点检测和配比 .结果表明 :(1 )应用 IPGphor,采用样品直接加入重泡胀溶液的形式 ,增大了溶解性 ,缩短聚焦时间、增大样品负荷量 (分析型 ) ,提高了分辨率 .(2 )比较宽 (p H=3～ 1 0 L)、窄 (p H=4～ 7L)范围 IPG胶条 ,窄 p H范围的 IPG胶条具有较高的分辨率 .(3)比较 PLA- 80 1 - C、D细胞蛋白图谱之间的差异 ,其相关系数为 0 .7339± 0 .0 2 91 ;仅在 PLA- 80 1 - C株出现的蛋白为 1 79个 .     

Proteomic study of human hepatocellular carcinoma using two-dimensional difference gel electrophoresis with saturation cysteine dye

To identify the proteomic alterations associated with carcinogenesis of hepatocellular carcinoma (HCC), we compared the protein expression profiles of nine HCC cell lines with those of primary cultured hepatocytes established from five individuals. A differential proteomic study was performed by two-dimensional difference gel electrophoresis, in which protein samples are labeled with different fluorescent dyes and separated according to their isoelectric point and molecular weight. To label the protein samples, we used a newly developed and highly sensitive fluorescent dye, which reacts with all reduced cysteine residues of proteins. Principal component analysis based on the intensity of 1238 protein spots indicated that the HCC cells and the normal hepatocytes had distinct proteomic profiles. The Wilcoxon test was used to determine the protein spots whose intensity was differentially regulated in the HCC cells compared with the normal hepatocytes, and mass spectrometric analysis was used to identify the proteins corresponding to the spots. The proteins identified are involved in cell cycle regulation, binding to a tumor-suppressor gene product, fatty acid binding, and regulation of translation. Western blotting with specific antibodies revealed the overexpression of PCNA, EB1 and E-FABP in HCC tissues compared with noncancerous tissues. Aberrant regulation of EB1 and E-FABP has not previously been implicated in the development of HCC.     

Susceptibility to oxidative stress: proteomic analysis of bronchoalveolar lavage from ozone-sensitive and ozone-resistant strains of mice

Previous studies have shown that the pulmonary response to ozone (O(3)) varies greatly among strains of mice, but the factor(s) and the mechanism(s) that are responsible for this differential susceptibility have not yet been clearly identified. The present study explores the molecular bases for this differential O(3) susceptibility by studying the expression of proteins associated to the epithelial lining fluid (ELF) from two strains of mice, C57BL/6J and the C3H/HeJ, respectively described as O(3)-sensitive and O(3)-resistant. The ELF proteins of these two strains were displayed by two-dimensional gel electrophoresis (2-DE) of bronchoalveolar lavage fluids (BALFs) and the protein patterns obtained with BALF samples of both strains were compared. Two major differences were observed between the BALF 2-DE protein maps obtained from C57BL/6J and C3H/HeJ strains. First, two isoforms of the antioxidant protein 2 (AOP2) were detected in a strain-dependent manner: C3J/HeJ possesses only AOP2a (isoelectric point 5.7) and C57BL/6J exhibits only AOP2b (isoelectric point 6.0). Second, the levels of anti-inflammatory and immunosuppressive Clara cell protein-16 (CC16) were 1.3 times higher in the BALF from resistant C3H/HeJ than from sensitive C57BL/6 mice. Moreover, two 6 kDa isoforms of CC16 with isoelectric points of 4.9 (CC16a) and 5.2 (CC16b) are detected in both strains. Interestingly, the C57BL/6J strain had a twice decreased level of the acidic isoform of CC16 compared to C3H/HeJ. Our results suggest that AOP2 and CC16 might participate in the protection of the pulmonary tract to O(3)-induced lung injury. The possible differential contribution of specific protein isoforms in the differential susceptibility to oxidative stress is discussed.     

A reference map of a human pituitary adenoma proteome

In order to compare the proteomes from different cell types of pituitary adenomas for our long-term goal to clarify the molecular mechanisms that participate in the formation of pituitary adenoma, and to detect any tumor-related marker for an "early-stage" diagnosis, the two-dimensional gel electrophoresis (2-DE) reference map of a pituitary adenoma tissue proteome is described here. A vertical, two-dimensional (2-D) polyacrylamide gel electrophoresis system and PDQuest image analysis software have been used to provide a high level of between-gel reproducibility and to accurately array each protein expressed in a pituitary adenoma tissue. Mass spectrometry (matrix-assisted laser desorption/ionization-time of flight MALDI-TOF and liquid chromatography-electrospray ionization-quadrupole-ion trap LC-ESI-Q-IT) and protein databases were used to characterize each protein in the 2-D gel. The results demonstrate that a good reproducibility of the 2-D gel pattern was attained. The position deviation of matched spots among four 2-D gels was 1.95 +/- 0.45 mm in the isoelectric focusing direction, and 1.70 +/- 0.53 mm in the sodium dodecyl sulfate-polyacrylamide gel electrophoresis direction. A total of ca. 1000 protein spots were separated by 2-DE, and 135 protein spots that represent 111 proteins were characterized with mass spectrometry (96 spots for MALDI-TOF, 39 spots for LC-ESI-Q-IT). The characterized proteins include pituitary hormones, cellular signals, enzymes, cellular-defense proteins, cell-structure proteins, transport proteins, etc. Those proteins were located in the cytoplasmic, cellular membrane, mitochondrial, endoplasmic reticulum, nuclear, ribonucleosome, extracellular fractions, or were secreted in plasma, etc. Those identified proteins contribute to a functional profile of the pituitary adenoma proteome. These data will be used to expand the proteome database of the human pituitary, which can be accessed in the website http://www.utmem.edu /proteomics.     

Proteomic analysis of protein expression profiles during Caenorhabditis elegans development using two-dimensional difference gel electrophoresis

Coordinated protein expression is critical for the normal execution of animal development. To obtain overall proteome profiles during animal development, a small free-living soil nematode, Caenorhabditis elegans, was used as a model and the developmental changes of protein expressions were analyzed using two-dimensional difference gel electrophoresis. Protein samples from six developmental stages were prelabeled with fluorescent cyanine dyes and separated on two-dimensional electrophoresis gels. Image-to-image analysis of protein abundances together with protein identification by peptide mass fingerprinting yielded the developmental expression profiles of 231 spots representing 165 proteins. About a quarter of the identified proteins were expressed in multiple spots with different isoelectric points, suggesting a certain proportion of proteins were variously modified. This notion was supported by the observation that about a third of the multispot proteins were stained positive for a phosphoprotein specific dye. While a fairly large number of the proteins showed little alteration in their expression profiles during development, about 40 proteins were found to be significantly either up- or down-regulated between the embryos and newly hatched L1 larvae. Down-regulated proteins included those related to the cell cycle such as MCM-7, PCN-1, and the mitotic checkpoint protein, while up-regulated proteins included structural proteins such as actins, LEV-11, DIM-1, VAB-21, metabolic enzymes such as ATP synthase, ALH-12, fluctose-1,6-bisphosphate aldolase and GPD-3, and galectins. A standard proteome map was obtained where the defects in the mutations of developmental genes and the effects of reagents on the development in C. elegans were analyzed.