CALR mutations are discovered in classic myeloproliferative neoplasms (MPN) as a new biomarker very recently.CALR mutations occur in about 20 ％-35 ％ essential thrombocythemia and primary myelofibrosis and are associated with some clinical features and favorable prognostic impact.This article reviews biological characteristics and clinical implications of CALR mutations in classic MPN.

Objective To investigate the relationship between JAK2 V617F mutation and vascular embolism diseases,in order to provide important basis for clinical diagnosis and treatment and prevention of embolism.Methods Patients who were hemoglobin ＞ 160 g/L,platelets ＞ 300×109/L treated in department of neurology,heart and vascular surgery in Xuanwu Hospital of Capital Medical University were collected.Vessel embolism and JAK2 V617F mutation situation and correlation were retrospectively analyzed.Results Among the total 56 cases,JAK2 V617F gene mutation positive rate was 37.50 ％ (21/56),the incidence of embolism was 40.07 ％ (23/56),there was correlation between JAK2 V617F mutation and embolism (P =0.014).Conclusion JAK2 V617F mutation is helpful to early diagnosis and treatment of myeloproliferative neoplasm,reduce thrombosis complication,improve the quality of life.

This article focuses on the rapidly evolving understanding of the molecular pathogenetic mechanisms of the bcr-abl-negative myeloproliferative neoplasms (MPN) [myeloproliferative disorders (MPD)],such as polycythemia vera (PV),essential thrombocythemia (ET),and primary myelofibrosis (MF).The amplify therapies were reviewed and IFN-α is an effective agent for these MPN (MPD).Also,the article emphasize once again avoidance MPN and select MPD for such chinese patients.

The outcome of allogeneic hematopoietic stem cell transplantation (allo-HSCT) has been improved over past 50 years due to the advances in HLA matching and increasing sources of HSC donors,such as developments and progressions of HLA-matched sibling donor transplantation (MSDT),umbilical cord transplantation (UCBT/CBT),unrelative donor transplantation (URDT),and HLA haploidentical transplantation (haplo-SCT).To review and discuss progressions in field of allo-HSCT,we studied relative advances reports of Education Program Book,54th-ASH,2012.Howeover,the outcomes of allo-HSCT can be quite different according to different diseases,disease phase or patient age.Furthermore,according to our local experiences,we emphasize again significance of HSCT donor safety.

Acute myeloid leukemia (AML) originates from rare leukemia stem cells (LSC).Because these chemotherapy-resistant LSC are thought to underlie disease relapse,effective therapeutic strategies specifically targeting these cells may be beneficial.With the increasing knowledge regarding the LSC,several major directions in targeted therapies with regard to LSC are being investigated.These include targeting cell surface molecules,signal pathways,and microenvironment of LSC.Since eliminating LSC-should provide an efficient,potentially curative treatment option for leukemia patients,this article reviews the recent progress of LSC targeting in AML therapy.

1 病历摘要 患者男,35岁,因"发现皮肤出血点4 d、加重2 d"于2011年2月3日入住首都医科大学附属北京朝阳医院.同年1月10患者因"上呼吸道感染"自行服用酚麻美敏片(泰诺)(2次共3片)、甲硝唑(2次4片)及牛黄类中药(2次3丸)等,此后五六天出现厌食、乏力等症状,未予重视.1月30日患者无诱因右上臂及小腿出现多发大小不等出血点,因抓挠扩大为瘀斑,同时出现右侧颊黏膜皮下血肿,遂到某医院就诊.血常规检查示血小板计数17×109/L,予中药(具体不详)后自行回家.2011年2月3日再次到该医院就诊,血常规检查示血小板计数4×109/L,丙氨酸转氨酶(ALT)1800 U/L.

This critical review was summarized more systematically about the JAK2V617F mutation of related research progress in myeloproliferative disorders (MPD) research fields and the identification of JAK2V617F mutation represents an important advance in our understanding of MPD was agreed. The authors focused on several sensitive problems of post the JAK2 mutation era, and expressed their opinions. The Guideline of the MPD diagnostic criteria recommended by WHO in 2008 was accepted. The authors recommend the MPD, rather than myeloproliferative neoplasm (MPN). The treatment for the MPD (not including the CML) is recommended. Before the effective targeting of JAK2V617F specific inhibitors for the treatment of the MPD, short-term of use hydroxyurea (HU) was suggested to suppress excessive proliferation of bone marrow of MPD and a long course of treatment application of inteferon-α(IFN-α), and low-dose of aspirin in a timely manner were recommended to prevent thrombosis and other complications.

人类白细胞抗原(Human Leukocyte Antigen,HLA),是一组展示在细胞表面分子,承担淋巴细胞识别和"抗原递呈"("antigen presentation").HLA通过识别"自己"和"非己"调控免疫反应,并因而成为移植排斥中的靶子.

目的探讨急性心肌梗死患者经冠状动脉自体骨髓单个核细胞治疗的安全性和对心功能的保护作用。方法2003年3月以来,84例急性心肌梗死患者急诊静脉溶栓或急诊PCI治疗后2周内行择期冠状动脉造影或PCI治疗。其中50例作为细胞治疗组,抽取骨髓40mL,提取单个核细胞,经冠状动脉注入;另34例作为对照组,经冠状动脉注入等量生理盐水。81例患者术前和术后6个月、2年行多巴酚丁胺负荷试验。29例治疗组患者和22例对照组患者术前和术后6个月行静态及动态心肌核素显像检查。结果细胞治疗组患者临床随访无明显副作用,心功能明显改善,运动耐力增加。多巴酚丁胺超声心动图负荷试验示左室射血分数（术前27.00%±0.89%,术后6个月36.80%±0.58%,术后2年40.94%±0.58%,术后6个月、2年与术前比,P均＜0.01）和室壁运动记分指数（术前1.55±0.05,术后6个月1.32±0.03,术后2年1.24±0.02,术后6个月、2年与术前比,P均＜0.01）显著改善,峰值射血分数（EF）与基础状态EF的差值（术前0.88%±0.54%,术后6个月15.06%±0.43%,术后6个月与术前比,P＜0.01）及WMSI和基础状态WMSI的差值（术前0.07±0.02,术后6个月0.19±0.02,术后2年0.15±0.01,术后6个月、2年与术前比,P均＜0.01）治疗前后之差异有统计学意义。动态与静态心肌核素显像提示梗死和缺血面积减小,存活心肌增加。结论经冠状动脉自体骨髓单个核细胞治疗的急性心肌梗死患者,经2年的临床观察无明显副作用,具备安全性,并显示出自体骨髓单个核细胞治疗对梗死后心功能有保护作用。

Objective To evaluate the clinical outcome of autologous hemopoietic stem cell transplantation(AHSCT) for hematological malignancies. Methods Data of 30 patients with hematological malignant diseases who under went AHSCT in Xuanwu hospital from January 1996 to January 2007 were retrospectively analyzed. There were 14 males and 16 females, the median age was 40(19~52) years. Twenty-five of them had acute non-lymphoblastic leukemias(ANLL)(CR1 24, CR2 1), 4 had acute lymophblastic leukemias(ALL)(CR1 4) and 1 had granulocytic sarcoma. Pretreatment regimens mainly included Mel 160 mg/m2+Ara-C 2.0 g/m2+CY 120 mg/kg or TBI 8~10 Gy+CY 120 mg/kg. Results All patients had rapid hemopoietic reconstitution. There was no AHSCT related death. The median follow up duration was 82(12~144) months. Twenty-one of 30 patients were still alive during the analysis. The probabilities of disease-free survival(DFS) at 5 years were significantly different in these two groups: (69±8)% for AHSCT groups and (34±10)% for synchronous intensive chemotherapy groups. Conclusion AHSCT can be safely performed as an important treatment constituent for hematological malignancies.

The existence of leukemia aberrant immunophenotypes (LAIP) has been suggested to be a valuable tool for the detection of minimal residual disease (MRD), as they could distinguish leukemic cells from normal hematopoietic progenitors. This study was purposed to analyze the characteristics of LAIP in acute leukemia and further explore the proportion of different types of LAIP in acute leukemia patients. Flow cytometry (FCM) with four color and CD45/SSC gating were used to detect the antigen expression in samples of bone marrow from 126 patients with acute leukemia. The results showed that definite LAIP could be detected in about 76% patients. The LAIP could be divided into four groups as cross-lineage antigen expression, asynchronous antigen expression, antigen overexpression and antigen lack expression. The percentages of these LAIPs were 39%, 46%, 21% and 29% respectively. About 11% out of analyzed cases showed the existence of only one aberrant phenotype while two or more of aberrant phenotypes could be detected in majority cases. It is concluded that the LAIP with four subgroups can be detected in the majority of patients with acute leukemia and immunophenotyping based on LAIP is applicable for the detection of MRD.

Objective:This study is to assess clinical safety of intracoronary transplantation of autologous bone marrow cells in patients with acute myocardial infarction(AMI),and to observe beneficial effect during the process of postinfarction remodeling.Method:There were 50 AMI patients randomly selected into bone marrow derived mononuclear in cells treatment group and 34 AMI patients in control group.All patients received emergency thromboysis or percutaneous transcoronary angioplasty(PTCA),and had patency of TIMI Ⅲ.During the next 14 days,patients received PTCA+stenting for coronary stenosis.Dobutamine stress echocardiography were performed before and 6-month,2-year after treatment.There were 40 ml aspirates of born marrow obtained in patients of cells treatment group.Mononuclear cells including endothelial progenitor cells were purified and counted,then transferred into infracted artery via 6F coronary angiography catheter.Result:Clinical follow up shows no side effect in 50 patients who received autologous bone marrow cells.Further more less nitroglycerin use and increased excise activity have been found in this group.Stress echocardiography shows significant improvement of left ventricular WMSI in treatment group patients.There were significant differences between peak WMSI and basement WMSI before and 6-month,2-year after cell treatment.Conclusion:Intracoronary autologous bone marrow cell transplantation is feasible and safey in patients with AMI.Stress echocardiography shows that autologous bone marrow cells transplantation plays a protective role in maintaining heart function after AMI.

Objective To evaluate the advantages and disadvantages of methods of ProCOUNT and single-platform ISHAGE gating strategy for CD34 positive cells in peripheral blood and apheresis samples. Methods Flow cytometry was adopted to enumerate CD34 positive cells in peripheral blood and apheresis samples with ProCOUNT and single-platform ISHAGE gating strategy. And enumeration of CD34 positive cells in 23 peripheral blood and 50 apheresis samples were performed. The results of the two methods were compared using Spearman correlation and paired t-test. Results There was a strong correlation between the absolute counting and percentage of CD34 positive cells for the two methods(r all ＞0.9, P all＜0.001). The absolute counting in peripheral blood and apheresis samples with ProCOUNT and single-platform ISHAGE gating strategy were (42.7±7.5)μL^(-1) and (52.1±7.2) (P＜0.01) respectively, (3515.2±684.7)μL^(-1) and (3923.8±711.1) μL^(-1) (P＜0.01) respectively. The percentage of CD34 positive cells in peripheral blood and apheresis samples were (0.26±0.07)% and (0.13±0.05)% (P＜0.01) respectively, (0.79±0.18)% and (0.83±0.17)% (P=0.056) respectively. All of the results using ProCOUNT were lower than those using single-platform ISHAGE gating strategy. Conclusion There is a strong correlation between the absolute counting and percentage of CD34 positive cells for the ProCOUNT and single-platform ISHAGE gating strategy, but the result of the former is significantly lower than that of the latter.

小李今年28岁,平时喜欢运动,身体特别壮,从小就没进过医院.今年参加单位体检时,医生发现他左颈部有1个"枣核"大小肿物,考虑是淋巴结肿大,建议他尽快进一步检查.

目的探讨成人急性髓系白血病(AML)多色流式细胞术免疫分型的特点.方法采用CD45/SSC双参数散点图设门方法进行多色流式细胞术免疫表型分析.结果髓系抗原CD13和CD33在AML各亚型中均有很高的表达,阳性率分别为98.3%和85.0%.CD14和CD64在AML-M4及AML-Ms患者中的表达率较高.CD14和CD64在AML-M4的表达率分别为60.0%和100.0%,在AML-Ms的表达率分别为80.0%和100.0%,但CD14的特异性要高于CD64.AML患者CD34、CD117、HLA-DR的表达率分别为59.3%、84.3%和71.7%;AML-M3患者CD117表达率为76.9%,CD34及HLA-DR呈低表达.AML患者淋系抗原CD7和CD19阳性率分别为20.0%和3.3%.结论采用CD45/SSC双参数散点图设门方法,使成人AML免疫分型结果更可靠.CD13、CD33、CD14可作为AML免疫分型的髓系单抗.CD34及HLA-DR低表达为AML-M3的特征.成人AML淋系抗原表达以CD7较常见.

目的了解间充质干细胞(mesenchymal stem cell,MSC)的培养条件、生物特性及分化能力.方法抽取健康成人骨髓,利用密度梯度离心分离培养人MSC,并用流式细胞仪检测细胞表面标记,体外诱导MSC向成骨细胞分化.结果成功地进行了人MSC的原代和传代培养;MSC CD34、CD45、HLA-DR、CD62p等为阴性,CD29、CD44、CD166、CD90等为阳性;MSC能够分化为成骨细胞.结论人骨髓MSC在体外有较强的扩增能力,并能够向成骨细胞分化.

OBJECTIVE:To compare the therapeutic effects of low-dose and high-dose interferon alpha-2b (IFN) treatment on chronic myelocytic leukemia (CML).METHODS:A real-time quantitative reverse transcriptase PCR (RQ-PCR) method was established to detect the fusion gene bcr-abl expression, thereby studying the reduction of leukemic cells. Thirty newly diagnosed CML patients, 21 males and 9 females, aged 14 - 69, were treated with hydroxyurea to keep the white blood cell count less than 20 x 10(9)/L, and then randomized into 2 groups: high-dose IFN group receiving IFN alpha-2b 5MIU 6 times per week for 3 - 6 months and low-dose IFN group receiving IFN alpha-2b 3MIU every other day for 3 - 6 months. Bone marrow was collected every month to Real-time PCR was used to detect the expression of bcr-abl mRNA. Mononuclear cells were isolated and RNA was extracted to detect the expression of fusion gene bcr-abl and a control gene GAPDH. The results were reported as the number of bcr-abl copies/GAPDH copy.RESULTS:The established real-time quantitative PCR method could detect the bcr-abl molecules as low as 50 copies. The intra-assay coefficient of variation (CV) was less than 5% and the inter-assay CV was 5.13%. The median bcr-abl fusion gene expression level of 30 CML patients before IFN therapy was 0.098 (range: 0.010 - 5.799). The bcr-abl expression level decreased by 19.37% and 24.86% in the low-dose and high-dose IFN groups respectively after 3 months' therapy. No significant difference was observed between the two groups (P = 0.398). Relatively more side effects were observed in the high-dose IFN group than in low-dose group.CONCLUSION:RQ-PCR is a reliable method to monitor CML therapy by analyzing fusion gene bcr-abl expression. There is a difference in bcr-abl fusion gene expression levels among the newly diagnosed patients, and low-dose IFN is as effective as high-dose IFN in reducing bcr-abl expression but with less side effects.

非甾体抗炎药(NSAIDs)引起的粒细胞缺乏症并不少见而且多起病急,病程进展快,如果得到及时有效的诊断和治疗,患者可以迅速康复,但是延迟诊断或误诊可使该病的死亡率升高.

乙型肝炎(乙肝)疫苗在新品种的开发、接种后无应答现象的基础研究方面,以及对免疫异常人群的预防、治疗的临床应用等方面取得了长足进步.介绍了三抗原重组疫苗Hepacare和脱氧核糖核酸(DNA)疫苗;乙肝疫苗在免疫功能异常患者中的应用;以及乙肝疫苗的治疗作用.

The aim was to study the roles that the bone marrow mesenchymal stem cells (MSC) and cytokines play in cord blood CD34(+) cell expansion ex vivo and the influence of culture ex vivo on expression of the adhesive molecule of CD44. CD34(+) cells sorted from cord blood cells had been cultured in each well of 24 well culture plates containing culture medium supplemented with mesenchymal stem cells layer or/and cytokines for a week, and then all kinds of indexes of different groups were compared. The results showed that as for cord blood cell expansion, there was no significant difference between the groups with cytokines SDF-1alpha + SCF + TPO + FL and SCF + TPO + FL no matter if MSC layer existed or not. The groups with MSC layer and cytokines were superior to the corresponding groups without MSC layer. In addition, the expression of the adhesion molecule CD44 had no distinct change after culture. It is concluded that SDF-1alpha has no distinct influence on the effect of cytokines SCF + TPO + FL on cord blood cell expansion ex vivo. MSC enhance the effect of cytokines on cord blood cell expansion ex vivo. Such expansion ex vivo may not influence the expression of the adhesive molecule CD44 on cord blood cells.