CD34、p53在脑多形性黄色星形细胞瘤及巨细胞胶质母细胞瘤中的表达及意义

目的：探讨CD34、p53在多形性黄色星形细胞瘤（PXA）及巨细胞胶质母细胞瘤（GCG）中的表达情况,以及两种肿瘤的临床病理特点、诊断及鉴别诊断。方法：对13例PXA及11例GCG进行临床病理学观察,CD34、p53等免疫组化分析。结果：PXA患者组织学特征： 肿瘤由巨怪瘤细胞、梭形细胞和泡沫样瘤细胞组成,有丰富的网状纤维及淋巴细胞浸润,坏死和核分裂象少见。GCG患者组织学特征： 瘤细胞多形性,以巨怪形瘤巨细胞为主,核分裂象和出血坏死多见,网状纤维沿血管周围分布,有淋巴细胞浸润。CD34在11例PXA中呈弥漫阳性表达,而在GCG中无一例阳性表达;p53在11例GCG中的阳性表达率达到60%-90%以上,而在PXA中的阳性率小于5%。结论：PXA及GCG临床、病理组织形态相似而预后完全不同,两者鉴别诊断的要点在于瘤细胞的异型性、核分裂象、坏死等。CD34、p53的免疫组化染色在两种肿瘤鉴别诊断中有重要意义。     

bcl-6、p53、c-myc基因异常在弥漫大B细胞淋巴瘤中的临床意义

目的 探讨bcl-6 、p53、c-myc基因异常的检测在弥漫大B细胞淋巴瘤（DLBCL）中的临床意义.方法 间期荧光原位杂交（I-FISH）方法检测59例DLBCL患者活体石蜡组织bcl-6、p53蛋白、c-myc基因异常的情况,同时以CHOP及R-CHOP方案化疗,评价疗效.观察bcl-6、p53蛋白、c-myc基因与化疗疗效及生存期的关系.结果 59例DLBCL中,p53丢失18例（30.5％）,bcl-6重排11例（18.6％）,c-myc重排5例（8.5％）.p53丢失阳性组化疗有效率（33.3％）明显低于阴性组（76.5％）（x2=9.560,P=0.002）. bcl-6基因重排阳性组的预后差于基因重排阴性组,但差异无统计学意义[总生存（OS）,P=0.107;无进展生存时间（PFS）,P=0.094]; p53基因丢失阳性组预后明显差于阴性组（OS,P=0.031;IPFS,P=0.028）;c-myc重排阳性组的预后差于基因重排阴性组,但差异无统计学意义（OS,P=0.163;PFS,P=0.167）.其中CHOP化疗组患者,p53基因丢失、c-myc重排阳性组的预后明显差于阴性组,差异有统计学意义（P值均＜ 0.05）;R-CHOP化疗组,bcl-6基因重排阳性组具有较差的预后意义（OS,P=0.003;PFS,P=0.007）.结论 bcl-6 、p53、c-myc基因异常与 DLBCL预后密切相关,可作为预测DLBCL的预后因素并指导治疗.     

PNH转MDS后急性粒细胞白血病变1例并文献复习

目的：观察阵发性睡眠性血红蛋白尿(PNH)向急性粒细胞白血病(AML)转化的克隆演变过程及其预后因素。方法：24岁男性患者因全血细胞减少而入院,经骨髓象、酸热糖溶血试验、免疫表型等辅助检查确诊为PNH。结果：诊断后2个月血片中出现原始细胞、骨髓病态造血;19个月后骨髓原始 早幼粒细胞升至0．53,确诊为AML。结论：PNH患者原始细胞表达CD34、CD117以及CD59阴性细胞进行性增高是不良预后因素;血栓栓塞是PNH的主要并发症及死亡原因。     

人不同来源CD+34造血细胞bcl-2、bax、Fas、Fas-L及p53的表达及意义

目的：探讨人不同来源正常CD34^ 造血干／祖细胞bcl-2,bax,Fas,Fas-L及p53的表达及意义。方法：分别收集了正常人骨髓、动员后外周血及脐带血,制备成单个核细胞后采用Mini MACS免疫磁珠分离法纯化CD34^ 造血干／祖细胞,再应用免疫组化SP法观察分离纯化的CD34^ 造血干／祖细胞bcl-2,bax,Fax,Fas-L及p53的表达。结果：所分离纯化的CD34^ 细胞经PACS分析,其纯度为≥98%,p53及Fas呈弱阳性或阴性,Fas-L及bcl-2呈强阴性或阳性染色,bax在脐血及骨髓为弱阳性,在外周血为阳性表达。结论：在不同来源的正常CD34^ 细胞bcl-2,bax,Fas,Fas-L及p53的表达相似。     

系统性肥大细胞增生症伴慢性粒-单核细胞白血病一例并文献复习

目的 应用细胞形态学、免疫学、细胞遗传学及分子生物学(MICM)诊断模式进一步认识系统性肥大细胞增生症伴相关血液肿瘤(SM-AHN).方法 对1例SM伴慢性粒-单核细胞白血病患者进行骨髓涂片观察计数原始细胞、异常肥大细胞比例.骨髓活组织检查观察阳性细胞分布及阳性程度;流式细胞术荧光标记设门分析;c-kit/D816突变检测;bcr-abl1融合基因检测;PDGFRA、PDGFRB基因重排检测、骨髓染色体核型分析.通过以上MICM检测结果分析并结合临床表现进行综合诊断,并进行文献复习.结果 该例患者骨髓涂片原始细胞0.06,肥大细胞0.26,POX原始细胞阴性,甲苯胺蓝肥大细胞阳性;可见淋巴样小巨核、单圆核巨核、双圆核巨核;骨髓活组织检查异形偏幼稚细胞致密灶状分布,呈梭形,比例为0.4～0.5,CD117(+)、CD25(+),CD2(-);骨髓流式细胞术提示原始、幼稚髓细胞0.02,免疫表型为CD34(+),CD117(部分+);CD117+CD25+CD2-的细胞约为10.8％;单核系列细胞约0.095,部分伴CD56异常;c-kit/D816突变阳性,bcr-abl1融合基因、PDGFRA、PDGFRB基因重排均阴性.结论 结合患者病史及MICM检查结果明确诊断SM-AHN,提示综合多学科检查结果诊断血液肿瘤具有重要意义.     

膀胱移行细胞癌中MDR、p53表达及细胞凋亡的意义

目的:探讨膀胱移行细胞癌(BTCC)中P-gP、p53基因及细胞凋亡表达的意义.方法:利用免疫组化S-P法和TUNEL法检测83例BTCC的P-gP、p53的表达及细胞凋亡指数(AI).结果:在83例膀胱移行细胞癌中P-gp阳性表达率为71.08%,p53阳性表达率为50.60%,AI为1.4335±0.3863;P-gp阳性表达在不同病理分级、临床分期及预后中无显著性差异(P＞0.05),p53及AI随着病理分级和临床分期而增高,均有显著性(P＜0.05),p53阳性表达复发组高于无复发组(P＜0.01),AJ复发组高于无复发组(P＜0.05).结论:检测P-gp可指导BTCC的化疗药物选择,p53高表达、细胞凋亡指数增高与BTCC的进展有关,P-gp、p53及AI高表达者预后及疗效差,p53可作为BTCC独立的预后因素.     

肠道非霍奇金淋巴瘤中的EB病毒感染和p21ras、p53蛋白表达及意义探讨

目的：研究肠道非霍奇金恶性淋巴瘤的EB病毒感染、p53、p21ras蛋白表达及其相关性。方法：应用单克隆抗体CD45RO)、CD20、CD78a、CD45RA、CD56、LCA、CD30、CD15、CD5、CD8、TIA-1、GramB、p53、p21；多克隆抗体CD3、κ、λ对瘤细胞进行免疫组化标记，用SABC免疫组化的方法检测瘤细胞p53、p21ras基因的表达及EB病毒寡核苷酸探针(EBER)原位杂交，观察了40例NHL的免疫表型、EBV感染情况及p53、p21ras蛋白的表达。结果：40例申，经免疫组化证实6例为T细胞淋巴瘤(15％)，34例为B细胞淋巴瘤(85％)。T细胞淋巴瘤为外周T细胞性和肠病型T细胞性淋巴瘤。B细胞淋巴瘤9例为弥漫性大细胞性淋巴瘤23.5％，25例为边缘区B细胞性淋巴瘤(76.5％)。EBV-ERBR原位杂交6／40例有阳性表达，均为T细胞淋巴瘤，阳性细胞占肿瘤细胞的30％～80％。P53的表达共有25例，21例有p21ras的表达。有16例同时检出p53和p21ras的表达。结论：肠道淋巴瘤以边缘区B细胞淋巴瘤多发临床为惰性。如为T细胞性淋巴瘤，更多见的是侵袭性，且T细胞淋巴瘤与EBV相关性较高B细胞淋巴瘤則无相关性。P53的表达与EBV感染无明显相关性，而p21ras的表达与EBV感染似有关系。     

乳腺叶状肿瘤临床病理特征分析

目的:探讨免疫表型在各级乳腺叶状肿瘤(PT)及纤维腺瘤(FA)鉴别诊断中的作用。方法:对85例PT和20例FA的临床病理资料进行回顾性分析,同时进行Bcl-2、CD117、CD34、p53和Ki-67免疫表型进行检测,比较观察两者组织学形态和免疫表型差异。结果:部分PT与FA形态相似,两者的免疫标志Bcl-2、CD117、CD34和p53表达有差异,χ2值分别为20.629、13.080、22.579和27.319,P值分别为0.001、0.004、0.001和0.001。Ki-67在FA、良性PT、交界性PT和恶性PT中平均增殖指数分别为1.50±1.100、3.02±5.629、8.00±8.582和21.92±17.916,F=19.776,P=0.001。Bcl-2、p53和Ki-67随着恶性程度增加表达增加,CD34则相反。结论:除了依据形态学以外,免疫组化Bcl-2、CD117、CD34、p53和Ki-67对鉴别诊断PT和FA有一定辅助诊断价值,临床可推广应用。     

伴T315I突变的Ph＋急性淋巴细胞白血病一例

患者 女,64岁.2013年2月19日因反复发热半月余,尿频、尿急10d入院.行骨髓穿刺检查分类：增生明显活跃,原始及幼稚淋巴细胞占0.815;免疫组织化学提示原始及幼稚淋巴细胞比例增高;流式细胞术分析异常细胞表达CD34、HLA-DR、CD10、CD19、CD33、CD38、CD123、TdT、cCD79a、CD22,弱表达CD20,不表达MPO、cIgM、sTgM;染色体核型为46,XX,t（9;22）（q34;q11）[10];MLL基因重排阴性;荧光原位杂交（FISH）检测bcr-abl融合基因阳性率90.4％,定量检测bcr-abl p 190121.52％;FLT-ITD及TKD突变阴性.     

胶质瘤中肿瘤干细胞的分布及与p53、MGMT的关系

目的：探讨胶质瘤中肿瘤干细胞标志物CD133的表达及其与p53、MGMT的关系。方法：采用免疫组化法检测45例不同病理级别胶质瘤标本中CD133和p53、MGMT蛋白的表达。结果：CD133在各病理级别的胶质瘤中均有表达,高恶性度组中CD133表达高于低恶性度组（P〈0.05）。在CD133阳性组中p53、MG-MT的表达明显高于CD133阴性组（P均〈0.05）。结论：不同恶性程度的胶质瘤标本中均存在肿瘤干细胞,肿瘤干细胞可作为判断胶质瘤恶性程度和预后的重要指标。     

Expression of hematopoietic progenitor cell associated antigen CD34 in chronic myeloid leukemia.

The expression of progenitor cell associated antigen CD34 was investigated in cells from 28 patients with chronic myeloid leukemia (CML). The CD34 positivity varied from 0-26% in patients with chronic phases CML (n = 17); from 6-64% in patients with accelerated phase CML (n = 4); and from 27-97% in the patients with blastic crisis of CML (n = 8). The difference in CD34 positivity between chronic (mean 10.1 +/- 2.3%), accelerated (37.7 +/- 13.3%) and blastic (58.0 +/- 7.3%) phases of CML is statistically significant (p less than 0.05), however, the number of patients studied, especially in accelerated and blastic phases is very small. There was no difference in the CD34 positivity of the cells in the peripheral blood and in the bone marrow. CD34 positivity was higher in patients with chronic phase CML at diagnosis (untreated patients) than in those who were studied during treatment. The possible importance of serially studying CD34 positivity in patients with CML is discussed in the paper.     

慢性粒细胞性白血病FLT3基因及FLT3/ITD基因突变的检测及意义

背景与目的:大部分急性髓细胞性白血病(acutemyeloidleukemia,AML)患者出现FLT3基因异常表达,20%～30%AML患者会出现FLT3/ITD基因突变并与临床预后相关。本研究旨在了解慢性粒细胞白血病(chronicmyeloidleukemia,CML)患者FLT3基因及FLT3/ITD基因突变情况。方法:采用聚合酶链反应(polymerasechainreaction,PCR)检测53例CML慢性期和34例CML加速期或急变期患者DNA水平FLT3基因及FLT3/ITD基因突变。结果:53例CML慢性期患者3例(5.7%)出现FLT3基因阳性,34例加速期或急变期患者有19例(55.9%)出现FLT3基因阳性,CML加速期和急变期患者FLT3基因阳性率显著高于慢性期患者(P<0.001),87例CML患者,仅2例(2.3%)出现FLT3/ITD基因突变。结论:在CML患者中,FLT3基因表达主要见于加速期或急变期患者;CML很少发生FLT3/ITD基因突变;FLT3基因及FLT3/ITD基因突变阳性CML患者可能提示预后不佳,此方面研究尚需深入。     

慢性粒细胞白血病p53基因功能检测

目的：通过以ｐ２１作为标志物的功能检测方法了解慢性粒细胞白血病中ｐ５３基因功能状况。方法：采用ｗｅｓｔｅｒｎ ｂｌｏｔ方法检测未经阿霉素诱导及阿霉素诱导后慢粒患者白血病细胞中ｐ５３及ｐ２１蛋白表达。结果;对２５例慢性患者白血病细胞进行了检测,显示５例ｐ５３基因功能异常,其中１例为慢性期,显示ｐ５３基因为野生型,但无功能;１例加速期;３例急变期,分别为急粒变,急单变和多克隆急变,在急淋变中未见。     

P53 Gene Mutations in Chronic Myelogenous Leukemia Medullary and Extramedullary Blast Crisis

Molecular alterations of the P53 gene were investigated in 27 unselected patients with chronic myelogenous leukemia (CML) blast crisis. A rearrangement of the P53 gene was evident by Southern blotting in 3 cases, one of which also showed the same alteration in the chronic phase. Single strand conformation polymorphism and sequencing analysis showed point mutations in 4 blast crisis cases. Of interest, P53 point mutations were evident in all the 3 cases of extramedullary blast crisis examined and the same point mutation was found in the myeloblastoma tissues and in the subsequent peripheral blast cells. These data indicate that: a) P53 gene mutations occur in a significant but not a large number of CML acute phase cases; b) P53 gene point mutations seem to correlate strongly with the infrequent extramedullary presentation of the blast crisis; c) the presence of the same P53 gene point mutation in extramedullary and bone marrow blast cells confirms the common clonal origin of the two cellular populations.     

Synergistic effects of p53 activation via MDM2 inhibition in combination with inhibition of Bcl-2 or Bcr-Abl in CD34+ proliferating and quiescent chronic myeloid leukemia blast crisis cells

The Bcr-Abl tyrosine kinase regulates several Bcl-2 family proteins that confer resistance to apoptosis in chronic myeloid leukemia (CML) cells. Given p53''s ability to modulate the expression and activity of Bcl-2 family members, we hypothesized that targeting Bcr-Abl, Bcl-2, and p53 concomitantly could have therapeutic benefits in blast crisis (BC) CML and in quiescent CML CD34⁺ cells that are insensitive to tyrosine kinase inhibitors (TKI). We examined the effects of the MDM2 inhibitor nutlin3a and its combination with the dual Bcl-2 and Bcl-xL inhibitor ABT-737, and the Bcr-Abl inhibitor nilotinib on BC CML patient samples. We found that in quiescent CD34⁺ progenitors, p53 expression is significantly lower, and MDM2 is higher, compared to their proliferating counterparts. Treatment with nutlin3a induced apoptosis in bulk and CD34⁺CD38⁻ cells, and in both proliferating and quiescent CD34⁺ progenitor CML cells. Nutlin3a synergized with ABT-737 and nilotinib, in part by inducing pro-apoptotic, and suppressing anti-apoptotic, Bcl-2 proteins. Nilotinib inhibited the expression of Bcl-xL and Mcl-1 in BC CML cells. These results demonstrate that p53 activation by MDM2 blockade can sensitize BC CML cells, including quiescent CD34⁺ cells, to Bcl-2 inhibitor- and TKI-induced apoptosis. This novel strategy could be useful in the therapy of BC CML.     

Frequency of expression of RHAMM/CD168 in Egyptian patients with CML

RHAMM/CD168 is a cell surface receptor for hyaluronan, a glycoaminoglycan that plays a fundamental role in cell growth, differentiation and motility. It is one of the leukemia-associated antigens (LAA) identified in patients with myeloid leukemias. WE AIMED: at studying the frequency of expression of RHAMM/CD168 in Egyptian patients with CML, both in chronic phase and accelerated/blastic phase, as a potential target structure for cellular immunotherapies, and to compare it with available western records. PATIENTS AND METHODS: RHAMM expression was tested by RT-PCR in peripheral blood mononuclear cells of 60 CML patients divided into 2 groups, group A: 44 chronic phase CML patients, group B: 16 accelerated/ blastic phase patients as well as 15 healthy volunteers. OUR RESULTS: Demonstrated that 14/44 (31.8%) of chronic CML patients showed positive RHAMM expression in contrast to 15/16 ( 93.7%) in the accelerated/blastic phase patients. Moreover within the chronic phase patients the RHAMM positive patients had a significantly higher level of bcr-abl/abl ratio. This highlighted the contribution of RHAMM expression with CML disease progression. CONCLUSION: Our work demonstrated a similar proportion of RHAMM expression in both Egyptian and western CML patients. This may pave the way for subsequent studies suggesting the concomitant use of RHAMM R3 peptide vaccination with conventional CML therapy especially in accelerated phase, in order to achieve complete molecular remission for our patient.     

Selective expression and constitutive phosphorylation of SHC proteins [corrected] in the CD34+ fraction of chronic myelogenous leukemias

The BCR/ABL fusion protein is a constitutively active tyrosine kinase that is responsible for the pathogenesis of chronic myelogenous leukemia (CML). Clinically, CML is characterized by a chronic phase (CP) that eventually terminates into a blast crisis (BC). BC transformation is associated with accumulation of CD34+ blasts. We investigated the expression and phosphorylation of Src-homology-2 and collagen-homology domains (SHC) [corrected] proteins in subpopulations of CML primary cells. Shc polypeptides are tyrosine kinase substrates that are constitutively tyrosine-phosphorylated in continuous cell lines of CML origin. High levels of Shc expression were found in the CD34+ cells from CML-BC, CML-CP and normal bone marrow. In contrast, CD34- fractions from CML-CP and normal bone marrow expressed low levels of p46Shc. Shc proteins were constitutively phosphorylated in the CD34+ fractions from CML cells (both CP and BC), but not in normal CD34+ cells. These data bear implications for the role of Shc in normal hemopoiesis and CML leukemogenesis: (a) dramatic changes of Shc expression during terminal differentiation of hemopoietic cells adds a further level of regulation to the signal transduction function of Shc; and (b) constitutive Shc tyrosine-phosphorylation in the rare CD34+ cells of CML-CP might contribute to the selection of this subpopulation during the blast crisis transformation of CMLs.     

STI571治疗慢性髓系白血病的临床疗效与毒副作用观察

背景与目的:bcr-abl融合基因翻译的蛋白产物P210bcr-abl的酪氨酸激酶(proteintyrosinekinase,PTK)活性异常增高被认为是导致慢性髓系白血病(chronicmyeloidleukmeia,CML)发病的根本原因。STI571能高效特异性抑制P210bcr-abl的PTK活性,在临床应用中获得了显著的疗效,但对急变期患者的治疗效果维持时间短。本研究观察和比较了STI571治疗慢性期与加速/急变期CML患者的临床疗效和所发生的不良反应,并从细胞遗传学的角度对急变期患者STI571耐药机制进行初步的分析。方法:选择接受STI571治疗的CML患者22例,其中慢性期6例,加速/急变期16例。按照血液学缓解和细胞遗传学缓解的标准,结合骨髓细胞形态学分析、骨髓细胞G显带技术分析和间期荧光原位杂交检测结果,对患者STI571治疗前和治疗3个月后的血液学和细胞遗传学缓解情况进行分析,并对3个月内出现耐药复发的患者进行核型演化分析。同时密切观察各系统发生的不良反应及严重程度。结果:6例(100%)慢性期CML患者获血液学完全缓解和细胞遗传学缓解,4例(25%)加速/急变期CML患者获血液学完全缓解,8例(50%)获不同程度的细胞遗传学反应。获血液学完全缓解和细胞遗传学反应的百分率两组比较均有统计学差异(P<0.05)。3例急变期CML患者出现耐药复发,其中2例可见2Ph和其它新     

Platelet Derived Growth Factor Expression, Myelofibrosis and Chronic Myelogenous Leukemia

CML is often associated with myelofibrosis, and fibrosis in the accelerated phase is one of the diagnostic criteria for this accelerated phase. In this review, the mechanism of myelofibrosis associated with CML is discussed with emphasis on the cell origin of the production and release of platelet derived growth factor (PDCF) and its interaction with marrow fibroblasts. In the initial stage of myelofibrosis in chronic phase CML, atypical small megakaryocytes might leak PDGF, possibly PDGF-AB, together with other growth factors. As the clinical phase of the disease progreses to accelerated or blastic phase, a larger quantity of PDGF-AB or PDGF-BB might be secreted from blastic cells with myeloid phenotype. In addition some fibroblasts may be attracted by the PDGF and proliferate, and deposit collagen as well as fibronectin in the bone marrow stroma.