目的 探讨以肿瘤性胸腔漏出液为主要表现的多发性骨髓瘤病例的临床特点.方法 报告解放军总医院血液科2015年收治的1例以肿瘤性胸腔漏出液为主要表现的多发性骨髓瘤病例,就其临床特点及诊疗经过结合相关文献进行回顾性分析.结果 患者男性,49岁,双侧胸腔积液导致活动后憋喘,无其他症状.入我院前曾就诊于多家大型医院,由于化验为胸腔漏出液且为唯一临床表现,均重点考虑良性疾病而未能找到病因,曾给予抗感染、抗结核治疗无效,对症给予胸腔置管引流缓解症状.每具自行引流胸腔积液1 500 ～3 000 ml,病程长达1年,多次复查血清白蛋白正常.入我院后查血清免疫固定电泳IgG-LAM阳性,提示可能为浆细胞疾病.骨髓涂片浆细胞占4.8％,免疫分型克隆性浆细胞占有核细胞的5.31％.但患者无骨髓瘤相关症状,不符合活动性多发性骨髓瘤诊断标准.且胸腔积液细胞学阴性.采用流式细胞术行胸腔积液免疫分型检查,发现克隆性浆细胞占有核细胞的49.99％,提示胸腔积液为克隆性浆细胞浸润所致,据此确诊为多发性骨髓瘤.结论 骨髓瘤性胸腔积液可以为漏出液,且可以为多发性骨髓瘤主要临床表现,但临床罕见.用流式细胞术对胸腔漏出液进行免疫分型检查可提高诊断准确率.

Objective To analyze the efficacy of a new iron chelator , deferasirox, in treatment of iron overload secondary to myelo-dysplastic syndrome ( MDS) and investigate its adverse reactions .Methods Clinical data of all the iron overload patients secondary to MDS who receiving deferasirox therapy in our hospital from January 2012 to April 2014 were collected and retrospectively analyzed . Their serum ferritin ( SF) level, amount of red blood cell transfusion , and hemoglobin level were measured and recorded before and after treatment, and adverse reactions were observed .Results There were totally 8 cases of MDS secondary iron overload enrolled in this study.They were 7 males and 1 female, and at a median age of 52 (ranging from 38 to 71) years.After 3 months’ treatment, complete response (CR) was obtained in 3 cases, minor response (MiR) in 3 cases, and stable iron load (SIL) in 2 cases.The overall response rate was 75.0%(6/8), and the median amount of red blood cell transfusion was 2 (1-3) u/month.In 1 year after treatment, SF level was significantly decreased [(871.0 ±584.2) vs (2164.9 ±1233.6) ng/ml], while that of hemoglobin was obviously increased[(101.5 ±34.59) vs (65.37 ±21.35) g/L], with statistical differences (P <0.05).At this time point, 5 patients were out of red blood cell transfusion , and the amount of red blood cell transfusion was 0.5, 1.5 and 2.0 u/month respectively, for the other 3 patients.After 1 year treatment, only 1 patient died, 3 patients experienced nausea and vomiting, and 1 patient diarrhea .Conclusion Deferasirox therapy is safe and effective for MDS secondary iron overload patients .

BACKGROUND:Post-transplant lymphoproliferative disease (PTLD) is a rare and serious complication after allogeneic hematopoietic stem cell transplantation (allo-HSCT) or solid organ transplantation.MATERIAL/METHODS:We conducted a retrospective analysis of the occurrence of post-transplant lymphoproliferative disease in allo-HSCT recipients over 12 years in a single center in China. A total of 343 patients received allo-HSCT. The conditioning therapy consisted of a busulfan/cyclophosphamide-based regimen, a fludarabine/cyclophosphamide-based regimen, or total-body irradiation and cyclophosphamide. In transplantations from unrelated donors and haplo-identical donors, patients also received antithymocyte globulin (ATG) or thymoglobulin as part of the conditioning.RESULTS:Five of the 343 patients (1.46%) were diagnosed with PTLD and all 5 were given ATG as part of conditioning. Among these 5 patients, 4 had lymphoid neoplasm before transplantation. EBV-positivity was confirmed in 4 patients. All 5 PTLD patients received reduction of immunosuppression (RI) as fundamental therapy. At follow-up on April 1, 2013, 1 patient had survived for 2 years and 1 had survived for 9 years. The correlation of PTLD with ATG and underlying diseases were examined by statistical analysis using the chi-squared test or Fisher's exact test (P=0.011 and 0.025, respectively).CONCLUSIONS:Although only 1.46% of patients progressed to PTLD associated with ATG and underlying diseases, the mortality was still high. Moreover, RI can be an effective therapy for PTLD patients, but other approaches should be further explored.

Spectral karyotyping (SKY) is a novel cytogenetic technique, has been developed to unambiguously display and identify all 24 humans chromosomes at one time without a priori knowledge of any abnormalities involved. SKY can discern the aberrations that can't be detected very well by conventional banding technique and Fluorescent in situ hybridization (FISH). So SKY is hyper accurate, hypersensitive, and hyper intuitionist. We will review the elements and application of SKY in leukemia.

Purpose of this study was to analyse the characteristics of clinical, iconographical, pathological and treatment methods of Langerhans cell histiocytosis (LCH), so as to improve the diagnosis and treatment level of this disease. The clinical data of 35 LCH patients were studied retrospectively. These patients were divided into 2 groups according to age <14 years old and ≥ 14 years old. The clinical symptoms were analysed and the signs, imageology and pathology manifestation and treatment results were evaluated. The results showed that LCH clinical manifestations were diverse and complex. Surgical treatment for patients with single system involvement of LCH was better than that of multi-system involvement of LCH (MS-LCH). For the latter, combined chemotherapy effects was better. After 3-year follow-up, 1-year OS was 94% ± 4%, 2-years OS was 91% ± 5%, 3-year OS was 86% ± 7%. 3 years OS of group <14 years old and ≥ 14 years old was 94% ± 6% and 81% ± 10% respectively. The OS of former was better than that of the later, but because a small number of cases, this difference was not statistically significant. It is concluded that LCH is easy to be misdiagnosed, the pathological biopsy is the gold standard of LCH diagnosis. The PET-CT can be of great help in identifying stages and finding lesion areas of the disease. Pulmonary Langerhans cell histiocytosis (PLCH) is more common in adult. Combined chemotherapy can improve the prognosis of the patients. The treatment methods should be choosed according to the stage and classification of disease.

Post-transplant lymphoproliferative disorder (PTLD) is a life-threatening complication for recipients of solid organ transplantation (SOT) and allogeneic hematopoietic stem cell transplantation (allo-HSCT). In SOT recipients, who previously have a normal immune system that has been immunosuppressed, reduction of immunosuppression (RI) shows favorable outcome. However, in HSCT recipients, who have been profoundly immunosuppressed and for whom the tempo of immune reconstitution cannot be fast enough to eliminate the lymphoproliferative process, RI is ineffective in most patients. Therefore, cases of tumor regression via RI alone are rare in the setting of HSCT. We present a case of 26-year-old female developing a polymorphic B-cell PTLD 4 months after receiving allo-HSCT for T cell lymphoblastic lymphoma. RI alone led to regression of the nasopharyngeal tumor, and no sign or evidence of graft-versus-host disease (GVHD) after RI was observed. The general condition of this patient was quite well just before we submitted our draft. To our knowledge, this is the first case that tumor of PTLD regressed upon RI alone with a favorable prognosis and without any evidence of GVHD and relapse of PTLD after RI therapy in the setting of HSCT, which justify the possible advantage of RI alone for low-risk patients.

This study was aimed to detect the expression of AML1 fusion genes in the patients with adult acute myeloid leukemia (AML) and further to investigate their association with the progression and prognosis of AML. Bone marrow samples were collected from 168 patients with de novo adult AML, and the expression of AML1 ETO, AML1-EVI1, AML1-MDS1, AML1-MTG16, AML1-PRDM16, AML1-LRP16, AML1-CLCA2 and AML1-PRDX4 was analyzed by a novel multiplex nested RT-PCR. Positive samples and minimal residual disease were further examined by real-time fluorescent quantitative PCR. The results showed that the AML1 fusion genes were found in 10.7% (18/168) patients. Among them, AML1-ETO in 12 (7.1%) cases were detected, AML1-EVI1 in 2 cases (1.2%), and AML1-MDS1, AML1-MTG16, AML1-PRDM16, and AML1-CLCA2 in 1 case (0.6%) each were detected. Among the patients with AML1-ETO, 10 patients (10/12, 83.33%) achieved complete remission (CR) after one cycle of chemotherapy, while 2 patients achieved CR after 2 cycles of chemotherapy. The 2 patients with AML1-EVI1 failed to achieve CR after one cycle of chemotherapy. Patients with AML1-MDS1, AML1-MTG16, AML1-PRDM16, or AML1-CACL2 did not achieve CR after one cycle of chemotherapy. It is concluded that AML1 fusion genes are more frequent and can provide the molecular markers for diagnostics and prognosis evaluation of AML and for monitoring MRD.

目的通过1例经克拉屈滨治疗的复发毛细胞白血病的病例分析，进一步了解毛细胞白血病的治疗策略和方法。方法观察2012年4月我院收治的1例经克拉屈滨治疗的毛细胞白血病复发患者的治疗过程，分析毛细胞白血病的临床特点、治疗方法、治疗效果及不良反应，并复习有关文献。结果此例患者经1个疗程克拉屈滨治疗，3个月后达到部分缓解，随访5个月持续部分缓解。结论克拉屈滨是治疗毛细胞白血病的有效方法，不良反应小，耐受性好，适合在中国毛细胞白血病患者中应用。

This study was purposed to investigate the clinical efficiencies and adverse reactions of treating the myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML) by using decitabine. The clinical data of 12 MDS and AML patients treated with decitabine were analyzed retrospectively. Among 12 patients there were 1 case of MDS-RA, 2 cases of MDS-RAEB-I, 3 cases of MDS-RAEB-II, 2 cases of AML-M4, 2 cases of AML-M5, 1 case of AML-M6 and 1 case of AML-M0. In decitabine chemotherapy program for 5 days (n = 8), decitabine 20 mg/(m(2)·d) × 5 days was applied, 4 weeks for 1 cycle; in program for 3 days (n = 2), decitabine 15 mg/m(2), once 8 h for 3 days, 6 weeks for 1 cycle; another program (n = 2), decitabine 20 mg/(m(2)·d) every other day for 5 times. For 1 patient achieved complete remission (CR) after treatment with decitabine, ID4 gene methylated level was detected by MS-PCR and ML-PCR before and after treatment. The results showed that 2 cases achieved CR, 1 case partial remission, 5 cases stable disease, 1 case progress of disease and 3 cases died. Disease control rate was 66.67% (8/12), the effective rate 25% (3/12). The average survival time was (11.5 ± 2.1) months. 1-year OS rate was 40%, 2-year OS rate was 16.7%. MS-PCR detection showed that the decitabine could significantly reduce the ID4 gene methylation level. It is concluded that decitabine can stabilize disease status of MDS patients, reduce blood transfusion dependence and improve the life quality of patients, and even some patients who transformed from MDS to leukemia achieved CR after treatment with decitabine. Decitabine can reduce the ID4 gene methylation level. The main adverse reaction of decitabine was myelosuppression, infection and so on. So the blood transfusions, antibiotics and other supportive treatments for these patients are needed. Most of patients well tolerate the adverse effects of decitabine after active symptomatic and supportive treatment. The efficacy and survival rate of patients in this study were similar to that of application of decitabine to treat MDS in other domestic studies.

Alteration in the balance between cell apoptosis and proliferation is one of the pathophysiological mechanisms of the myelodysplastic syndromes (MDS). The question of whether the excessive apoptosis and/or proliferation predominantly involve the subset of progenitor cells (CD34(+) cells) or mature cells (CD34(-) cells) remains a controversial issue. This study was purpose to analyze the apoptosis and proliferation status of CD34(+) and CD34(-) cells in bone marrow (BM) of patients with MDS, to investigate the pathogenesis of MDS and to determine the relation of apoptosis and proliferation status of CD34(+) and CD34(-) cells with prognosis of MDS. The proportion of CD34(+) cells, the apoptosis and proliferation ratio (A/P) of CD34(+) and CD34(-) cells in BM of 40 patients with MDS, including 20 cases of high-risk MDS and 20 cases of low-risk MDS, and 10 normal persons as control were detected by flow cytometry; the influence of CD34(+) and CD34(-) cell apoptosis and proliferation levels on prognosis of MDS was evaluated by univariate and multivariate analysis of survival. The results showed that the proportion of CD34(+) cells in BM of high-risk MDS patients was significantly higher than that in BM of low-risk MDS patients and in normal BM [(1.92 ± 0.10)%, (1.09 ± 0.04)%, (1.03 ± 0.05)% respectively]. The apoptotic rates (AR) of both CD34(+) and CD34(-) cells were significantly higher in low-risk MDS [(54.75 ± 2.18)%, (80.36 ± 1.68)%] than in high-risk MDS [(24.87 ± 2.69)%, (23.12 ± 1.23)%] and in normal BM [(18.51 ± 2.74)%, (20.98 ± 2.21)%]. When compared between CD34(+) cells and CD34(-) cells in low-risk MDS, a greater AR of CD34(-) cells was found. However, the higher proliferative rate of CD34(+) cells was observed in high-risk MDS. In low-risk MDS, a higher A/P ratio was found in CD34(-) cells than in CD34(+) cells; whereas this ratio was equalized or inverted in high-risk MDS. In addition, the survival and prognosis correlated significantly with AR of CD34(+) cells. It is concluded that the early MDS is predominantly associated with excessive apoptosis of the mature CD34(-) cells. The proliferation rate of cells increases with the disease progression in MDS subsets, especially, in the subset of CD34(+) cells. Surprisingly, the apoptosis of CD34(+) cells may be a useful prognostic factor, and the inhibition of apoptotic mechanisms may induce the transformation of MDS to leukemia.

OBJECTIVE:To explore the role of SHP-1 promoter methylation on the pathogenesis and progression in myelodysplastic syndromes (MDS) and its related mechanism.METHODS:63 MDS patients were divided into low-grade (LG) group and high-grade (HG) group according to IPSS score system. Bone marrow samples were collected. Methylation specific-PCR (MSP) were used to detect the status of SHP-1 promoter methylation in bone marrow (BM) samples from different risk MDS patients and MDS cell line, SKK-1. Western blot was used to detect signal transduction and activator of transcription (STAT3) activation in SKK-1 cell line and MDS patients.RESULTS:No SHP-1 promoter methylation could be detected in healthy controls BM. Partially methylation was found in SKK-1 cell line. Methylation rate of SHP-1 gene promoter was found in BM of 24.2% of low-grade MDS patients and 63.3% of high-grade MDS patients, the difference between these two groups was statistically significant (P < 0.05); Patients were divided into different groups according to WHO subtype, chromosomal karyotype and blast cells in bone marrow, methylation rates of SHP-1 were significantly higher in RAEB-II, poor karyotype group and samples with 0.11-0.19 blast cells (P < 0.05); The phosphorylation protein of STAT3 was detected in SKK-1 cell line. The expression of phosphorylation STAT3 was significantly higher in HG group than in LG group (66.7% vs 18.2%) (P < 0.05). There was a significant correlation between SHP-1 promoter methylation and STAT3 phosphorylation.CONCLUSION:Abnormal methylation of SHP-1 gene promoter might have tentative role in the pathogenesis and progression of MDS, which may be involved in STAT3 activation. Detection of SHP-1 promoter methylation may be helpful to evaluate the prognosis of MDS.

Introduction Decitabine to treatment of patients with MDS into AML patients achieved complete remission during the treatment,in order to learn about the efficacy and side effects of anti-tumor drugs DNA methylation inhibitor Decitabine.Decitabine for MDS patients can significantly improve the quality of life,reduce transfusion dependence,even able to make the transformation to leukemia patients achieved complete remission with the side effect of bone marrow suppression.And a comprehensive review of the literatures is made to Decitabine clinical application in recent years,to provide help for the clinical application of the medicine.

Objective To improve the understanding of Langerhans cell histiocytosis(LCH).Methods Clinical features of 1 LCH patient were reported,including high fever,bone pain,cough,pleural effusion,multiple liver and spleen occupying lesions.PETCT showed that bones,spleen,mediastinum,hepatic lymph nodes were extensively involved.Right iliac bone biopsy revealed that the tumor cells were histocytes with their cytoplasm red stained and the chromatin structure was loose with visible nucleoli.Immunohistochemistry showed S100 + + +,CD1a + +,and CD68 +,which were classified as a high-risk group with multiple systems involved.Results The clinical symptoms of the patient did not improve after one cycle of CHOP chemotherapy.Her bone pain disappeared and her body temperature became normal after one cycle of ICE chemotherapy.The clinical symptoms disappeared after 6 cycles of ICE chemotherapy.Reexamination showed that the bone marrow and positron emission tomography(PET-CT) returned to normal.The patient was followed up for 8 months during which her symptoms were completely alleviated.Conclusion Biopsy is the golden diagnosis standard for LCH and positive immunohistochemical S100 and CD1a are the pathological characteristic of LCH.PET-CT does contribute to its definite staging,measurement of its size and assessment of its clinical outcome.ICE combined chemotherapy can achieve good clinical outcomes when multiple systems are involved.

ETV6 is an important hematopoietic regulatory factor and ETV6 gene rearrangement is involved in a wide variety of hematological malignancies. In this study, we sought to investigate the incidence of ETV6-associated fusion genes in B- and T-lineage acute lymphoblastic leukemia (ALL) by multiplex-nested reverse transcription-polymerase chain reaction (RT-PCR) in 176 adult ALL patients. Total RNA was extracted from bone marrow samples of ALL patients including 136 B- and 40 T-lineage ALL, and ETV6 fusion genes were detected by multiplex-nested RT-PCR. Changes of ETV6 fusion gene mRNA transcript levels were examined by real-time RT-PCR. We detected a total of 15 ETV6 gene rearrangements with a positive rate of 8.5%, involving seven ETV6-associated fusion genes in 13 B-ALL (13/136, 9.6%) and 2 T-ALL patients (2/40, 5.0%). ETV6-RUNX1 were observed in six cases (3.4%), ETV6-JAK2 in three cases (1.7%), ETV6-ABL1 in two cases (1.1%), and ETV6-ABL2, ETV6-NCOA2, ETV6-SYK, and PAX5-ETV6 each in one case (0.6%). ETV6-JAK2 was found in both B-ALL and T-ALL patients. Furthermore, real-time quantitative RT-PCR assays showed that the ETV6-RUNX1 mRNA transcript levels decreased during conventional chemotherapy or hematopoietic stem cell transplantation. This study shows that multiplex-nested RT-PCR is an effective and accurate tool to identify ETV6 rearrangements in adult ALL, which provides some clues into the diagnosis and prognosis of ALL but also molecular markers for the detection of minimal residual disease in adult ALL.

Objective To study the clinical characteristics of multiple myeloma with double clones and analyze its related laboratory diagnostic and therapeutic methods.Methods A case of multiple myeloma with double clones of IgG-κ and IgG-λ was reported and its related literature was reviewed.Results The myeloma protein ingredients were different between multiple myeloma with double clones and typical multiple myeloma.Immune fixation electrophoresis was of diagnostic value for multiple myeloma with double clones.The treatment outcome was usually a decreased or disappeared myeloma protein ingredient.The prognosis of patients was rather good.Conclusion Multiple myeloma with double clones is a rare plasma cell disorder,which is different from typical multiple myeloma in clinical manifestations,diagnosis,treatment and prognosis,and should thus be differentially diagnosed from typical multiple myeloma.

Objective To study the application of dual color-dual fusion fluorescence in situ hybridization(DC-DF-FISH) in detection of gene rearrangement in adult acute leukemia(AL) patients.Methods AML1/ETO,MLL,PML/RARa,BCR/ABL gene rearrangement was detected by DC-DF-FISH in 26 adult AL patients and compared with that detected by routine R-band.Results Four marker chromosomes were detected in 26 adult AL patients with a positive rate of 15.4%.No abnormal chromosome containing 11q23 was found in 17 patients with normal or abnormal karyotypes.Positive specific marker genes were detected in 8 patients by DC-DF-FISH and R-band respectively,with a positive rate of 30.8%.Conclusion DC-DF-FISH can reliably detect the PML/RARa,MLL,and AML1/ETO gene rearrangement,and can thus be used in diagnosis of AL,in judgment of therapeutic effect,and in detection of small residual foci.

This study was aimed investigate the recombination event occurring between HLA-A and-A loci discovered from father's HLA haplotype chromosome in a family. Peripheral blood samples were collected from a family. HLA class I (-A, -B, and -Cw) and II (-DRB1 and -DQB1) alleles were amplified and typed by both low and high resolution PCR with sequence-specific primers (PCR-SSP) and sequence-based typing (SBT). The results showed that 2 haplotypes of the patient were A(*)3001-B(*)1302-DRB1(*)0701 and A(*)3001-B(*)5601-DRB1(*)1454 respectively, those of her father were A(*)3001-B(*)1302-DRB1*0701 and A(*)1101-B(*)5601-DRB1(*)1454. Family analysis demonstrated that the patient's A(*)3001-B(*)1302-DRB1(*)0701 came from her mother and A(*)1101-B(*)5601-DRB1(*)1454 came from her father, but the A of patient was A(*)3001 and B, DR were the same to her father. This showed that the chromosome exchange and recombination event of father's 2 haplotypes occurring between HLA-A and -A loci at meiosis. And recombinate haploid chromosome was completely inherited to his daughter 1. HLA typing and Paternity testing demonstrated that father was the natural father, and the recombination event occurring between HLA-A and -A loci of the daughter 1 with father's HLA haplotype chromosome. It is concluded that the HLA-A/A of father's HLA haplotype chromosome recombination event occurring between HLA-A an-A loci has been found in a family in China, which helps further study on the mechanisms of HLA recombination.

OBJECTIVE:To evaluate the value of spectral karyotyping (SKY) in cytogenetic analysis of acute myeloid leukemias (AML).METHODS:Nine AML patients were analyzed by R-banding and SKY. MLL, PML-RARalpha, AML1-ETO fusion genes were detected by dual fusion- fluorescence in situ hybridization (D-FISH).RESULTS:All 9 samples were successfully hybridized. SKY identified structural aberrations including 9q -, t(15;17) and ins(10;17) (q22;p11p12) ; and some numeral abnormalities. The results of SKY confirmed those of R-band karyotyping and D-FISH; with more accurate localization.CONCLUSION:SKY appears to be fairly stable, accurate and sensitive, for AML cytogenetic study.