骨髓活检对小儿再生障碍性贫血的诊断价值探讨

目前,虽然对再生障碍性贫血(简称再障)的诊断已有国内和国外的统一标准,但仍有相当一部份不典型病人难以作出明确诊断,我院儿科近5年对32例再障患儿进行了骨髓活检,现将结果分析如下。 资料和方法 一、对象 40例确诊为再障的患儿进行了骨髓活检,其中8例因取材不佳未能得出结果,取材成功率为80％。在取材成功的32例患儿中,男17例,女15例,年龄5岁～13岁,急性再障10例,慢性再障22例,诊断标准按1987年全国再障学术会议制定的诊断标准。对照组:选择9例非血液系     

再生障碍性贫血患儿骨髓祖细胞培养的临床意义

目的探讨再生障碍性贫血(AA)患儿红细胞集落生成单位(CFU-E)、爆式红细胞集落生成单位(BFU-E)、粒细胞-单核细胞集落生成单位(CFU-GM)及巨核细胞集落生成单位(CFU-Meg)骨髓祖细胞培养情况,了解骨髓祖细胞在AA发病中的作用及意义。方法抽取28例AA患儿骨髓3～4mL,以淋巴细胞分离液提取单个核细胞,用磷酸盐缓冲液(PBS)洗涤2次,调节单个核细胞水平为106/mL,取0.3mL分别接种于红系、粒-单核系、巨核系培养基分别作4类祖细胞培养,在7、14d测定其集落数。结果28例AA中,4类集落数均值与对照组比较,差异有显著意义(P<0.01或P<0.001)。集落数随病情加重而逐步下降。4类集落数低于对照组患儿下限所占百分比分别为BFU-E35.71%、CFU-E85.71%、CFU-GM75.00%、CFU-Meg89.29%。治愈及进步者4类集落数接近对照组水平,治疗无效及进展者与治疗前比较无变化。结论动态监测AA患儿骨髓4类祖细胞集落数变化对辅助诊断、观察疗效及预后判断均有指导意义。     

骨髓活检在小儿再生障碍性贫血临床应用的研究

为探讨骨髓活检在小儿再生障碍性贫血（简称再障）临床应用的价值，采用骨髓活检改良制片法对88例再障患儿进行骨髓组织学定量分析并与涂片对照；同时对其中30例进行治疗后观察；并对76例进行随访。结果表明：小儿再障诊断符合率和疗效判定骨髓活检高于涂片检查，疗效与骨髓造血组织容量，造血细胞绝对值呈正相关，而与非造血细胞绝对值呈负相关；初检造血组织容量大小与生存期长短有关。结果提示：骨髓活检改良制片法适用于儿童，其可作为小儿再障早期诊断、疗效判定、预后评估的一个客观指标。     

骨髓活检在小儿再生障碍性贫血临床应用的研究

为探讨骨髓活检在小儿再生障碍性贫血（简称再障）临床应用的价值，采用骨髓活检改良制片法对８８例再障患儿进行骨髓组织学定量分析并与涂片对照；同时对其中３０例进行治疗后观察；并对７６例进行随访。结果表明：小儿再障诊断符合率和疗效判定骨髓活检高于涂片检查，疗效与骨髓造血组织容量，造血细胞绝对值呈正相关，而与非造血细胞绝对值呈负相关；初检造血组织容量大小与生存期长短有关。结果提示：骨髓活检改良制片法适用于儿     

再生障碍性贫血患儿骨髓核素显像特点及其临床意义分析

目的 应用99mTc-硫胶体全身骨髓显像检测再生障碍性贫血（再障）患儿全身骨髓显影特点,探讨其在再障诊治中的价值。方法　对2009年1月至2012年1月青岛大学医学院附属医院小儿血液科收治的45例再障患儿（再障组）及15名对照组进行99mTc-硫胶体全身骨髓显像。99mTc-硫胶体注射放射剂量10～15 MBq/kg,分析骨髓核素显像的特异性、敏感性及不同类型再障患儿骨髓显像特点和临床转归的相关性。结果　再障组45例骨髓显像中活性减低33例（73.3%,包括0级11例、1级22例）,表现为全身骨髓显像有不同程度受抑,其特点是全身骨髓显影不良、显影骨髓总量减少;2级12例（26.7%）,骨髓活性正常。45例再障组临床诊断为急性再障（SAA）组19例、慢性再障（CAA）组26例,其中SAA组骨髓显像表现为中央＋外周骨髓均抑制者15例,仅中央骨髓抑制2例;CAA组中央＋外周骨髓均抑制6例,仅中央周骨髓抑制6例,仅外周抑制4例;两组骨髓显像抑制部位差异有统计学意义（χ2= 10.37,P＜0.05）;提示慢性再障骨髓显像受抑程度较轻,可仅有中央或外周骨髓的抑制并多存在灶状增生。对再障患儿骨髓显像与早期治疗反应进行统计分析,结果示不同骨髓显像分级间疗效差异有统计学意义（χ2=12.49,P＜0.05）,骨髓显像0级组有效率为36.3%,较1级组、2级组（77.3%、83.4%）疗效差。结论　全身骨髓显影不良、显影骨髓总量减少及显像特点有助于对儿童再障的诊断、分型及预后判断。骨髓显像与临床转归和预后关系仍需进一步扩大病例研究。     

再生障碍性贫血患儿骨髓间充质干细胞分离及培养

目的 分离再生障碍性贫血(再障)患儿骨髓间充质干细胞,进行体外培养,为进一步研究打下实验基础.方法 从再障患儿及正常志愿者骨髓中分离骨髓间充质干细胞,进行体外培养,诱导向成骨细胞、成脂细胞分化和流式细胞仪检测表面标志CD34、CD45、CD73和CD90.绘制生长曲线描述原代和传代细胞生长情况.结果 分离的贴壁细胞经向成骨细胞和脂肪细胞分化诱导培养,分别显示茜素红染色和油红染色阳性;CD73和CD90阳性.生长曲线显示,与正常志愿者相比再障患儿体外培养的骨髓间充质干细胞传代培养中生长能力减低.结论 再障患儿骨髓间充质干细胞存在异常,可能在再障发病机制中起着重要作用.     

再生障碍性贫血患儿骨髓造血干细胞端粒酶活性及相关基因表达的研究

目的：探讨再生障碍性贫血（简称再障）患儿骨髓造血干细胞端粒酶活性及端粒酶逆转录酶（hTERT）基因和端粒酶RNA组分（hTERC）基因的表达变化和关系。方法：用重复序列扩增（TRAP）银染法和实时荧光定量RT-PCR法分别检测52例慢性再障患儿（CAA组）、13例急性再障患儿（SAA组）和21例骨髓造血正常儿童（对照组）端粒酶活性及hTERT和hTERC mRNA的表达。结果：CAA组、SAA组端粒酶活性和hTERT mRNA水平均高于对照组（P<0.01）,且CAA组端粒酶活性和hTERT mRNA水平较SAA组升高（P<0.05）;hTERC mRNA在各组间表达差异无统计学意义（P=0.812）;hTERTmRNA的表达水平与端粒酶活性呈正相关（r＝0.660,P＜0.01）。结论：端粒酶活性的表达变化与儿童再障发病及发展过程有关,hTERT表达在端粒酶活性调节过程中起重要作用。     

小儿再生障碍性贫血死因分析

目的：了解小儿再障的死亡原因,从而努力降低死亡率。方法：采用回顾性调查方法对住院时死亡１８例,院外死亡７６例共９４例小儿再障进行死因分析。结果：１８例急重羡慕 障住院时死亡的原因中,死于感染１１例占６１．１１％,死于出血７例占３８．８９％;７６例院外死亡者,死于出血４３例占５６．５８％,死于感染１０例占１３．２９％,其它死因５例,死因未查明１８例。结论：感染与出血是小儿再障最常见最严重的并发症,而     

儿童再生障碍性贫血骨髓组织超微结构观察及其临床意义

目的　探讨儿童再生障碍性贫血(简称再障)骨髓组织超微结构临床意义。方法　采用透射电镜对35 例儿童再障的骨髓组织进行超微结构观察,并与3 例骨髓造血功能正常的其他血液病骨髓组织进行对照。结果　儿童再障骨髓造血细胞和基质细胞均有不同程度改变,造血细胞改变程度慢再障高于严重再障,基质细胞改变程度严重再障高于慢再障;临床缓解和治愈的再障骨髓组织仍与正常不同。结论　再障患儿骨髓组织本身存在的内在缺陷是再障发病、药物不能根治、停药易复发的基本原因,再障骨髓超微结构的变化特征对于指导再障诊断的分型、治疗有一定的临床意义     

Hemogram and bone marrow morphology in children with chronic aplastic anemia and myelodysplastic syndrome

Background  Aplastic anemia (AA) and myelodysplastic syndrome (MDS) are both acquired disorders in which bone marrow fails to produce or release sufficient blood cells. Anemia, infections and thrombocytopenia are common signs of such diseases. Clinically, it is difficult to distinguish chronic aplastic anemia (CAA) from MDS, especially from MDS without splenomegaly. As prognosis and treatment of AA and MDS are different, it is extremely important to make a differential diagnosis for the two diseases. Methods  The medical records of 31 patients with CAA and 17 patients with MDS were retrospectively reviewed. Hemogram, bone marrow smear and biopsy for those patients were analyzed. Results  The mean counts of monocytes and platelets in the peripheral blood of the CAA patients were significantly lower than those of the MDS patients. Bone marrow smear showed a reduction of cellularity in CAA patients. The mean counts of myeloblasts+promyelocytes, myeloblasts+proerythroblasts, and megakaryocytes in the bone marrow of CAA patients were markedly lower than those in MDS patients. But the mean lymphocyte count was reversed. Bone marrow cells showed morphological abnormalities in MDS. Hematopoietic tissue in the bone marrow biopsy decreased obviously in more than 96% of the patients with CAA. Adipose tissue in the bone marrow of CAA patients increased obviously. A reduction or deficiency (<2 cell/piece) of megakaryocytes was noted in 28 patients with CAA. Fibrous tissue in the bone marrow was detected in 5 patients with CAA. Bone marrow biopsy results showed hypercellular changes in 12 MDS patients. Ten patients showed aggregated erythroblasts which were in the same stage of development, and 15 patients had abnormal localization of immature precursors (ALIP). Conclusions  Blood cell counts can be decreased in addition to the reduction of cellularity in the bone marrow without dyshematopoiesis in CAA patients. Peripheral blood monocytes, fibrous tissue and cellularity in bone marrow are increased in MDS. Dyshematopoiesis and ALIP may appear characteristically in the children with MDS. Histology of bone marrow is important in the differential diagnosis of MDS and CAA.     

Successful bone marrow transplantation for children with aplastic anemia based on a best‐available evidence strategy

Okamoto Y, Kodama Y, Nishikawa T, Yamaki Y, Mougi H, Masamoto I, Tanabe T, Shinkoda Y, Kawano Y. Successful bone marrow transplantation for children with aplastic anemia based on a best‐available evidence strategy.
Pediatr Transplantation 2010: 14:980–985. © 2010 John Wiley & Sons A/S. Abstract: A best‐available evidence strategy, i.e., the best‐available donors, conditioning regimens and GVHD prophylaxis were chosen at the time of BMT for AA, was analyzed retrospectively. The outcomes for 18 children with AA who underwent allogeneic BMT were analyzed. The median age was 11 yr (range 4–16), and nine were men. As conditioning regimens, seven had low‐dose irradiation + CY, six had ATG + CY + Flu, and five had ATG + CY. Donors were HLA‐matched siblings in 10, HLA‐mismatched family in one, HLA‐matched unrelated in three, and HLA‐mismatched unrelated in four. As GVHD prophylaxis, three received CsA alone, nine received CsA + MTX, and six received tacrolimus + MTX. All 18 patients showed engraftment. The median number of days until the neutrophil count exceeded 500/μL was 16 (range 11–21) post‐transplant. Five developed more than grade 2 acute GVHD, and three developed extensive cGVHD. One patient died because of interstitial pneumonia complicated with cGVHD. Five‐yr OS was 94% (95% CI: 83–105). These results suggest that a strategy of treating patients based on the best‐available evidence is acceptable.     

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??Abstract?? Objective To investigate the characteristics of 99mTcmsulfur collid bone marrow scintigraphy in childhood aplastic anemia ??AA?? and its clinical significance.Methods A total of 45 children were newly diagnosed with AA??including 19 cases of serious aplastic anemia ??SAA?? and 26 cases of chronic aplastic anemia ??CAA??.Bone marrow scintigraphy using 99mTcmsulfur collid 10??15 MBq/kg was performed in 45 patients and 15 controls.The difference of bone marrow scintigraphy was analysed. The relationship between laboratory indices and clinical response was investigated. Results According to the bone marrow imaging results??grade 2 of marrow activities was defined as normal. Totally 33 patients??33/45??73.3%?? had abnormal marrow activities??and in 15 controls only 3??3/15??20%??.Most AA patients showed 99mTcmsulfur collid uptake decrease. CAA patients had higher marrow activities than SAA patients ??P??0.05??.In the 33 patients who had abnromal marrow activities??less then 2 grade????SAA and CAA patients showed different 99mTcmsulfur collid uptake in the peripheral and the center of bone marrow??and CAA had focals whose uptake was increasing.After treatment of 6 months??45 AA patients were investigated the relationship between the grade of marrow activities and the therapeutic effect??including improved??inefficacy??death??.There showed significantly difference??P??0.05?? between them. Conclusion The 99mTcmsulfur collid bone marrow imaging is helpful to the diagnosis and prognosis of AA??but further study is needed to judge its relationship with the curative effect and prognosis.     

Low serum lipids suggest severe bone marrow failure in children with aplastic anemia

BACKGROUND: Significantly low serum lipid levels are occasionally seen at the time of diagnosis in children with aplastic anemia (AA). The aim of the present study was to clarify the pathologic and clinical significance of pretreatment serum lipid levels in AA. METHODS: A questionnaire seeking precise data about AA in pediatric patients, including the initial laboratory data at the time of onset of AA and the clinical course of these patients, was sent to 18 institutes in Japan; 13 institutes responded to the questionnaire. In this retrospective study, data concerning hematologic examination and serum lipids were available for analysis in 127 children with AA. Serum lipoprotein patterns were analyzed using conventional agarose electrophoresis in eight patients. In order to elucidate the cause of hypolipidemia in AA, we assayed serum macrophage colony stimulating factor (M-CSF), which is well known to have apparent cholesterol-lowering activity, by using an enzyme-linked immunosorbent assay in seven patients with hypocholesterolemia and compared the results with those obtained in patients with iron-deficiency anemia (IDA). RESULTS: We found that pretreatment total cholesterol (TC) and triglyceride levels in the serum correlated well with counts of both nucleated cells and hemopoietic cells in the bone marrow (BM) and were inversely correlated with the lymphocyte ratio in both the BM and peripheral blood. Patients with serum TC lower than 150 mg/dL showed a poor response to any form of therapy except BM transplantation. There was no difference in the serum lipoprotein patterns between the controls and patients examined. The serum M-CSF level was significantly higher in patients with TC levels lower than 150 mg/dL compared with controls. CONCLUSIONS: These results indicate that the pretreatment serum lipid level may reflect hematopoietic activity within the BM and can help to predict the therapeutic response of each case of AA to treatment with immunosuppressive drugs, corticosteroids and anabolic steroids. These results also indicate that M-CSF may be one of the contributing causes of the hypocholesterolemia that occurs in both AA and IDA.     

Outcome of children with aplastic anemia treated with immunosuppressive therapy

Background

Immunosuppressive therapy (IST) is the alternative treatment in children with aplastic anemia (AA) who do not have an HLA‐matched sibling. The aim of this study is to evaluate the outcome of children with AA treated with IST.

Methods

We retrospectively reviewed the hospital records of children with AA from 1984 to 2004, treated at our institution with antithymocyte globulin (ATG), cyclosporine (CS), and short course of prednisone.

Result

Forty‐two patients were treated with IST (24 boys, 18 girls); of whom 26% received G‐CSF. The median age at diagnosis was 8.5 years. Sixty‐nine, 19, and 12% were diagnosed with severe, very severe, and moderate AA, respectively. Twenty‐one percent had hepatitis‐associated AA. Median follow‐up time was 53.3 months. Sixty‐two percent had complete response; 19% had partial response. Two patients relapsed and received a second course of ATG; both had a partial response. The actuarial 5 years survival rate was 67.5%. Two patients developed myelodysplastic syndrome (MDS); both received long‐term G‐CSF and had partial response after two courses of IST. Fifteen percent of survivors had significant hypertension which persisted after CS was discontinued.

Conclusions

This study shows promising response in children with AA treated with IST; however, the outcome was inferior to our institutional results with hematopoietic stem cell transplantation from a sibling donor. Hypertension and MDS are late complications. Longer follow‐up, larger cohorts, and prospective studies are warranted to evaluate late complications and risk factors. Pediatr Blood Cancer 2008;50:52–57. © 2007 Wiley‐Liss, Inc.     

Immunosuppressive treatment of aplastic anemia in Chinese children with antithymocyte globulin and cyclosporine

Fifty-one children with aplastic anemia (AA) from 1993 to 2004 in the authors' institution were treated by 3 therapies: 11 patients in group 1 received the SSL-6 protocol; 16 patients in group 2 had CsA alone, where the dose of CsA began from 9-12 mg/kg in the initial 2 weeks and tapered off to 5 mg/kg later; 24 patients in group 3 were treated combining rabbit ATG (Pasteur, Merieux) 2.5 mg/kg for 5 days with CsA, which was the same dose as in group 2. The response was 27, 50, and 79%, respectively. The statistical analysis showed that the protocol of intensive immunosuppressive treatment (IST) was the most effective one and CsA was better than that of SSL-6. None of our patients developed clone diseases although the follow-up was as long as more than to 9 years. The data suggest that children with AA should receive IST as first-line therapy in developing countries.

Hematopoietic stem cell transplantation (HSCT) is effective treatment for patients with aplastic anemia (AA). However, HSCT is not widely used in China for economic reasons and lack of donors. Immunosuppressive therapy (IST) is now the mainstay treatment for AA. To evaluate the effect of immunosuppressive therapy, combining antithymocyte globulin (ATG) with cyclosporine (CsA), a retrospective study on 51 children with AA from January 1993 to December 2004 treated in the authors' department was performed.     

儿童再生障碍性贫血的规范诊治现状及研究进展

再生障碍性贫血(再障)是儿童期严重的血液病之一,诊断和治疗较为困难,尤其是重型再障预后较差.我国属于再障高发地区,必须引起足够的重视.目前已经明确,T淋巴细胞"免疫介导"是再障的主要发病机制,由于T淋巴细胞功能异常,而导致骨髓造血干细胞受到抑制.成功的异基因造血干细胞移植虽能根治再障,但受到供体来源困难、医疗费用昂贵、治疗难度大和医疗风险高等限制,国内难以广泛开展.近年来,根据再障免疫介导致病机制所开展的免疫抑制治疗已经获得较为满意的疗效,是无合适供体进行造血干细胞移植的重型和难治型再障的最佳替代疗法.文章对有关再障发病机制的研究进展、诊断与分型标准、鉴别诊断要点、治疗原则与治疗方法以及疗效评价标准等进行简要介绍.     

Successful engraftment of haploidentical bone marrow with post‐transplantation cyclophosphamide in patients with aplastic anemia

Patients with severe aplastic anemia (SAA) may benefit from hematopoietic stem cell transplantation, but many of them lack a matched donor. Haploidentical transplantation is increasingly utilized for the treatment of nonmalignant disease where patients lack a matched donor. We report patients with aplastic anemia who experienced successful engraftments of haploidentical stem cells with post‐transplantation cyclophosphamide (PTCy). Case series and review of the literature. We present two cases of pediatric patients with severe aplastic anemia who experienced successful engraftment of haploidentical related bone marrow. Both patients received conditioning consisting of rabbit ATG, cyclophosphamide, fludarabine, and total body irradiation pretransplant, with PTCy. The conditioning regimen was well tolerated by both patients, and they achieved full donor engraftment and were weaned off all immunosuppressants. Haploidentical stem cell transplantation in patients with severe aplastic anemia may be an effective alternative when fully matched donors are not available. PTCy can facilitate successful engraftment and therefore expand the pool of eligible donors for patients with aplastic anemia.     

骨髓间充质干细胞缺陷与获得性再生障碍性贫血

获得性再生障碍性贫血(AA)患者骨髓间充质干细胞(BM-MSCs)缺陷已成为近年来研究的热点之一.该文对BM-MSCs 缺陷在AA 发病中的作用及其对AA 治疗的临床应用进行综述.越来越多的实验室证据证明了BM-MSCs 缺陷在AA 的发病中极有可能起着重要的作用,无论是其生物学特点、基因表达谱的缺陷,抑或是增殖分化能力、造血支持作用乃至免疫调节功能的耗竭,都成为在免疫失衡基础上促使AA 不断进展至难以恢复的重要节点.随着MSCs 研究的不断深化,将恢复骨髓造血微环境为主要目的的MSCs 输注可能成为AA 治疗的新模式.