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To explore the correlation between the changes of T lymphocyte subsets and cytokines in patients with MM and immune function status, biochemical indicators, and their relationships with clinical stage and prognosis, which is expected to provide a scientific basis for the prognosis analysis and condition monitoring of MM patients.The clinical data of 89 MM patients in two hospitals were collected, and 36 healthy people without tumor or infectious diseases were selected as the control group. Flow cytometry and enzyme-linked immunosorbent assay (ELISA) were used to detect the changes of core members of peripheral blood T lymphocyte subsets and cytokine levels, respectively. At the same time, automatic biochemical analyzer and automatic blood cell analyzer were used to detect serum βThe proportions of NK cells and CD8The abnormal regulation of the core members of T lymphocyte subsets and the levels of various cytokines are closely related to the disease progression and poor prognosis of MM patients, which is an effective indicator for the disease monitoring of MM patients.T细胞亚群与细胞因子水平变化在多发性骨髓瘤患者临床诊疗及预后评估中的价值.探讨多发性骨髓瘤（MM）患者T淋巴细胞亚群及细胞因子的变化与免疫功能状态、生化指标的相关性以及与临床病情阶段及预后的关系，为MM患者预后分析及病情监测提供科学依据.收集两家医院共89例MM患者的临床资料，同时选取36例健康者为对照组，采用流式细胞术、酶联免疫吸附试验（ELISA）分别检测入组患者外周血T淋巴细胞亚群核心成员及细胞因子的水平，同时应用全自动生化分析仪与全自动血细胞分析仪检测血清βMM患者外周血中NK细胞、CD8T淋巴细胞亚群核心成员及各细胞因子水平异常与MM患者病情进展、预后不良密切相关，是MM患者病情监测的有效指标.

To observe the safety of donor NK cell infusions in the settings of hematopoietic stem cell transplantation and after consolidation chemotherapy in elderly patients with acute myeloid leukemia (AML).Forty patients with AML were included, in which 21 patients aged over 60 years were at the stage of complete remission (CR) and 19 patients that received allogeneic hematopoietic stem cell transplantation (allo-HSCT). Mononucleated cells were isolated from peripheral blood from the donors (for allo-HSCT) or healthy immediate family members (elderly AML). The cells were seeded into the flasks pre-coated with NK cell specific activators, and expanded in media containing recombinant human IL-15 and IL-2 for 14 days. The cells were transfused intravenously after the identification of quality control. Trypan blue exclusion test was used for the determination of cell viability and counting. Flow cytometry analysis was performed to assess the surface antigenic profile. Seventy-eight infusions of the cell products were received by the elderly patients with AML after consolidation chemotherapy, 11 infusions were received by the patients during allo-HSCT and 32 infusions 3 moths after transplantation. The safety of cell therapy, body temperature, blood pressure and other indexes were observe during and 48 hours after cell transfusion. Meanwhile, the occurrence and severity of acute graft-versus-host disease (GVHD) were documented.Flow cytometry analysis showed that the proportion of NK cells (CD3NK cell products with a high-purity could be obtained by ex vivo expansion with this protocol. The transfusion of these expanded cells is generally safe in the elderly patients with AML that have received chemotherapy or patients that received hematopoietic stem cell transplantation.NK细胞预防移植复发及治疗老年白血病临床安全性观察.观察急性髓性白血病（AML）患者异基因造血干细胞移植（allo-HSCT）时、移植后及老年AML患者巩固化疗后，供者NK细胞输注的安全性。.入组40例AML患者，其中年龄60岁AML完全缓解（CR）患者21 例，allo-HSCT患者19例。采集健康直系亲属或供者外周血，密度梯度离心法分离单个核细胞，将细胞接种于NK细胞激活剂包被的培养瓶中，以重组人IL-15及IL-2细胞因子组合，优势扩增人NK细胞14 d。质检合格后进行静脉输注。台盼蓝拒染法计数细胞总数和活力，流式细胞术分析免疫细胞表型。AML患者巩固化疗后接受NK细胞输注，共78次；allo-HSCT患者移植时输注NK细胞11次，移植后3个月内输注NK细胞共32次。输注期间及之后48 h，记录患者体温、血压等，以观察细胞治疗的急性毒性，并记录急性移植物抗宿主病（GVHD）发生情况。.免疫表型分析显示，细胞扩增前外周血中NK细胞（CD3-CD56+）占单个核细胞的比例为（14.10±4.22）%（n=121），培养14 d后CD3-CD56+ 细胞比例为（87.29±8.75）%（n=121），NK细胞增殖倍数为 753.47±140.13倍（n=121）。NK细胞输注量为每次（7.58± 2.50）×10采用该体系可获得高纯度的NK细胞，用于白血病造血干细胞移植及老年AML患者，细胞输注是安全的。.

To investigate the clinical characteristics, etiology, therapy and outcome of hyperthyroidism after allogeneic hematopoietic stem cell transplantation (HSCT).The clinical data of 7 patients who experienced hyperthyroidism were retrospectively analyzed in our hospital.These 7 patients (5 males, 2 females) suffered hyperthyroidism after HSCT. All patients did not apply the pretreatment regimen containing total body irradiation (TBI). The median age was 25 years old, only one child. Six patients underwent haploidentical HSCT except one patient after unrelated HSCT. The median time of hyperthyroidism occurrence was 20 months. Two patients experienced chronic graft versus host disease (GVHD) when hyperthyroidism occurred and were treated successfully with glucocorticoid, however one patient suffered hypothyroidism 3 months later and needed long-term oral levothyroxine maintenance. One patient developed hypothyroidism post treatment ofHyperthyroidism is a rare complication after HSCT but may affect healthy and lead to lower quality of life. Routine thyroid function monitoring should be recommended after HSCT. Treatment of hyperthyroidism should be given according to the pathogeny.异基因造血干细胞移植术后并发甲状腺功能亢进的临床分析.探讨异基因造血干细胞移植术后并发甲状腺功能亢进的临床特点、发病机制、治疗及预后情况.回顾性分析中国人民解放军空军特色医学中心7例异基因造血干细胞移植术后并发甲状腺功能亢进患者的临床资料.7例患者中，男5例，女2例，中位年龄为25（7-32）岁，儿童1例，其余6例为成人，急性白血病3例，慢性粒细胞白血病1例，骨髓增生异常综合征1例，重型再生障碍性贫血2例。6例患者接受单倍体相合造血干细胞移植，1例接受无关供者全相合造血干细胞移植。所有患者均未应用含全身照射（TBI）的预处理方案，发生甲状腺功能亢进中位时间为20（9-25）个月。2例患者在诊断甲亢时合并慢性移植物抗宿主病，经激素及抗排异治疗后，甲亢得到控制，但其中1例在3个月后出现甲状腺功能减退症状，需要长期口服左甲状腺素维持。1例应用异基因造血干细胞移植术后甲状腺功能亢进是少见并发症，但严重影响患者生活质量，移植后长期存活患者需定期监测甲状腺功能，治疗上需根据引起甲状腺功能亢进的原因进行.

To analyze the clinical characteristics of SAA patients with post-transplantation lymphoproliferative disease (PTLD) after allogeneic hematopoietic stem cell transplantation, and to improve diagnosis and treatment of PTLD.The clinical data of 192 patients with SAA patients who underwent HSCT in a single center from September 2010 to September 2017 were analyzed retrospectively. All patients were received antithymocyte globulin（ATG） conditioning regimen and mesenchymal stem cell(MSC) infusion.Among 192 cases, PTLD occurred in 14 cases, the incidence was 7.29％， 9 of them were diagnosed by pathology， and 5 were diagnosed clinically. EBV infection occurred with a median time of 72（35-168） days, Viral load higher than 1×10The incidence of PTLD after HSCT with SAA who used ATG and MSC in conditioning regimen closely relates to EBV infection, age of patients≤12 years, HLA-mismatch in URD-HSCT, grade II to IV GVHD. Rituximab combined with RIS may reduce the incidence of PTLD, combined EBV-CTL and chemotherapy may be the useful and most important treatment for PTLD.重型再生障碍性贫血患者造血干细胞移植治疗后淋巴组织增殖性疾病的临床分析.分析重型再生障碍性贫血(SAA)患者造血干细胞移植（HSCT）后淋巴组织增殖性疾病(PTLD)的临床特征,以提高对PTLD的诊疗水平.回顾分析2010年9月至2017年9月在单中心行HSCT的192例SAA患者的临床资料，分析PTLD的有效预警和治疗方法。本组患者预处理方案中均含有ATG并接受间充质干细胞（MSC）输注.192例SAA患者HSCT移植后有14例并发了EBV+ PTLD，总体发生率为7.29%，其中临床诊断5例，病理诊断9例，移植后PTLD发生中位时间72(35-168)d,所有PTLD均发生了EBV感染，且EBV高载量大于1×10SAA患者接受预处理含有ATG和MSC的HSCT后PTLD发生与EBV高拷贝密切相关，年龄≤12岁儿童、非血缘HSCT中供者为HLA不全相合的移植及合并有II度以上aGVHD者移植后PTLD高发； RIS/RTX抢先治疗有效，疗效不佳患者及时应用CTL及联合化疗是控制病情有效及重要方法.

To analyze the prognostic factors of haplo-HSCT combined with MSC in the treatment of SAA.127 SAA patients who had undergone haplo-HSCT with co-infusion of MSC in our center from January 2014 to August 2019 were analyzed retrospectively. Median age was 11 (1-37) years, and median follow-up time was 39.8 (1-74) months.The median time for neutrophil and platelet engraftment was 14 d and 18 d respectively. The cumulative incidences of grade III-IV aGVHD was 4.4%±1.9% at day +100. The 2-year cumulative incidence of extensive cGVHD was 8.3%± 2.7%. The estimated 3-year OS was 86.1%±3.1%. Univariate analysis showed that high-dose CD34In the haplo-HSCT of SAA, the prognosis of children patients is better than that of adults patients. More CD34单倍体造血干细胞移植联合间充质干细胞治疗重型再生障碍性贫血.分析单倍体造血干细胞移植（haplo-HSCT）联合间充质干细胞治疗重型再生障碍性贫血（SAA）预后的影响因素.回顾性分析2014年1月至2019年8月在空军特色医学中心接受haplo-HSCT联合间充质干细胞输注治疗的SAA患者共127例，中位年龄11（1-37）岁，中位随访时间为39.8（1-74）个月.粒系和血小板的平均植入时间分别为14和18 d。+100 d III-IV度急性移植物抗宿主病（aGVHD）的累积发生率为4.4%±1.9%，2年广泛性慢性移植物抗宿主病（cGVHD）的累积发生率为8.3%±2.7%，3年总体生存（OS）为86.1%±3.1%。单因素分析结果显示：高剂量的CD34在SAA的haplo-HSCT中，儿童患者的预后明显优于成人，输注较多的CD34

To analyze the prognostic factors of AML children with CBFβ/MYH11 positive.Twenty-eight children with CBFβ/MYH11 positive treated in our hospital from May 2012 to June 2018 were selected, the clinical data and curative were analyzed and evaluated.Five-year OS and 5-year EFS rate of CBFβ/MYH11 positive AML children was 76.8% and 64.0% efficacy, respectively. Univariate analysis results showed that the OS rate of CBFβ/MYH11 positive AML children with WBC60.0×10WBC level and XRCC-Thr241Met genotype at initial diagnosis are the major affecting factors for prognosis of AML children with CBFβ/MYH11 positive.伴CBFβ/MYH11阳性初诊AML患儿的预后影响因素分析.分析伴CBFβ/MYH11阳性初诊急性髓系白血病（AML）患儿的预后影响因素.选取2012年5月至2018年6月本院收治的原发性初治伴inv（16）/ CBFβ-MYH11阳性AML患儿28例，对其临床资料及治疗效果等进行分析和评估.所有患儿5年总生存（OS）率76.8%，5年无事件生存（EFS）率64.0%。单因素分析结果显示，WBC60.0×10初诊WBC水平及XRCC-Thr241Met基因型为影响伴CBFβ/MYH11阳性初诊AML患儿预后的主要因素.

To analyze the therapeutic efficacy of haploidentical-hematopoietic stem cell transplantation (hi-HSCT) for patients with juvenile myelomonocytic leukemia (JMML).The engraftment of hematopoietic stem cells, incidence of graft versus host disease (GVHD), infection, relapse, and survival of 6 JMML patients received hi-HSCT were retrospectively analyzed.Six (6 males) JMML patients received hi-HSCT from haplo-HLA-matched related donors. The results showed that the hematopoictic stem cells in all 6 patients were grafted successfully. Two cases of JMML died of pulmenary infections, other 4 cases survive without disease. Acute GVHD occurred in 3 patients and chronic GVHD occurred in 1 patients. The overall survival, disease free survival and relapse rates were 66.7%, 66.7%, 0%, respectively.The hi-HSCT is an effective method for treatment of patients with JMML, but there also is a serial problems to be resolved.

To analyse the efficacy and safety of co-transplantation of umbilical cord mesenchymal stem cell(UC-MSC) with haploidentical hematopoietic stem cell transplantation(hi-HSCT) in children with hematologic malignancy.The clinical data of 47 children undergoing hi-HSCT were retrospectively analyzed from November 2003 to November 2014, among them 34 patients received UC-MSC from October 2011 to November 2014, and another 13 patients without UC-MSC from November 2003 to September 2011. The median follow-up time was 20(0.5-67) months.No adverse events were observed after the UC-MSC transplantation. The engraftment rate, the median neutrophils engraftment time and platelet engraftment time all were not significantly different between hi-HSCT and hi-HSCT+UC-MSCT(P0.05). The three-years cumulative overall survival (70.6% vs 23.1%),(P=0.004), three-years cumulative disease-free survival(52.9% vs 0) (P=0), and early cytomegalovirus (CMV) viremia (91.2% vs 38.5%) (P=0) in UC-MSC+hi-HSCT group were statistically significantly higher than that in the conventional hi-HSCT group. The morbidity of aGVHD (44.1% vs 92.3%) (P=0.003), I-II aGVHD (26.5% vs 61.5%) (P=0.041) and transplantation-related mortality (11.8% vs 46.2%) (P=0.017) in UC-MSC+hi-HSCT group was statistically significantly lower than that in hi-HSCT group, however, the morbidity of III-IV aGVHD (17.6% vs 30.8%), cGVHD (26.5% vs 30.8%), HC (35.3% vs 7.7%), pulmonary infection (52.9% vs 46.2%) and relapse rate (32.4% vs 53.8%) were not statistically significantly different (P0.05) between the 2 groups.The application of umbilical cord mesenchymal stem cell in children undergoing hi-HSCT is safe, the UC-MSC can improve the overall survival, disease-free survival and reduce transplantation-related mortality. UC-MSC can reduce the morbidity of aGVHD, but increase the early infection of CMV, however it is nothing for the pulmonary infection and relapse in the children after hi-HSCT.

To analyze the risk factors of patients with relapsed leukemia after allogeneic hematopoietic stem cell transplantation, and to explore the therapeutic strategies for recurrence.The Cox proportional hazard regression model was used for univariate and multivariate analysis of transplantation-related index, a single center retrospective study of clinical data of 202 cases of leukemia received allo-HSCT from March 2004 to October 2014 had been conducted to screen the risk factors for recurrence after transplantation.In the leukemia patients received allo-HSCT, 68 cases relapsed. The relapse rate was 33.6%. The median time of relapse was 4(1.5-26 ) months. Univariate analysis indicated that there were 5 risk factors related with the disease relapse(P0.05), including the type of disease, extramedullary disease prior to transplant, the course of induced remission, the status of disease at HSCT and chronic graft versus host disease(cGVHD). Multivariate analysis showed that extramedullary disease prior to transplant(RR=2.622, 95%CI 1.139-6.037), the course of induced remission(RR=1.156, 95%CI 0.682-1.957), cGVHD (RR=1.728,95%CI 0.999-2.991) were independent risk factors for relapse of the patients received transplantation. Treatment strategies for the relapsed patients included withdraw immunosuppressant, donor lymphocyte infusion, systemic chemotherapy and local radiotherapy, targeted therapy, and second transplantation. Individualized choice was needed according to the relapsed site. The relapse-related mortality was 25.2%.The relapsed patients with leukemia after allo-HSCT have poor prognosis, early interference has good effect. The evaluation and prevention of risk factors before transplantation is even more important.

To investigate the clinical difference of cytomegalovirus (CMV) infection between HLA-matched allogeneic hematopoietic stem cell transplantation (allo-HSCT) and haploidentical hematopoietic stem cell transplantation (hi-HSCT).The clinical data of 83 patients who had undergone allo-HSCT were retrospectively analyzed. Out of them 50 patients underwent hi-HSCT and 33 patients received grafts from HLA-matched donors. The sera of all recipients and donors were CMV-negative before transplantation. All patients accepted myeloablative regimen without total body irradiation. PCR was performed to detect CMV in the peripheral blood twice a week after neutrophil recovery. CMV-DNA500 copies/ml was defined as CMV viremia.68 patients (81.9%) were diagnosed as CMV viremia before 100 days after transplantation. The incidence of CMV infection in hi-HSCT group was 90% and significantly higher than that in HLA-matched HSCT group (69.7%) (P0.05). All the patients responded well to anti-CMV therapy; however, 63 cases experienced CMV reactivation. The occurrence rate of CMV reactivation in hi-HSCT group (95.6%) was comparable to that in HLA-matched HSCT group (87.0%) (P0.05). Univariate analysis showed that the transplantation pattern, the recovery time of peripheral neutrophils and the occurrence of acute graft-versus-host disease (aGVHD) significantly related to the episode of CMV viremia, while the sex and age of the recipients, and the recovery time of platelets did not associate with the incidence. Further analysis found that the recovery time of neutropils and platelets in HLA-matched HSCT group were greatly shorter than those in hi-HSCT group (P0.05). The incidence of aGVHD was comparable between two groups however, incidence of severe aGVHD was significantly higher in hi-HSCT (P0.05).The hi-HSCT is more susceptible to CMV infection, which may be related to the higher incidence of severe aGVHD and the relative delay of hematopoietic reconstruction as compared with HLA-matched HSCT.

To explore the safety and efficiency of unrelated donor peripheral blood stem cells (URD-PBSC) transplantation combined with umbilical cord mesenchymal stem cells (UC-MSC).The clinical data of 49 patients received unrelated donor peripheral blood stem cells transplantation (URD-PBSCT) for treating hematologic malignancies were retrospectively evaluated, including 12 ANLL, 17 ALL, 18 CML and 2 MDS. Out of them, 22 patients received the URD-PBSCT combined with UC-MSC and 27 patients received only URD-PBSCT. The average number of infusing UC-MSC was 1.0 × 10⁶/kg in the UC-MSC+URD-PBSCT group.As compared with URD-PBSCT group, in UC-MSC+URD-PBSCT group the median recovery time of neutrophilc granulocytes was shorter (12 d vs 15 d) (P = 0.041), the incidence and severity of chronic graft versus host disease (cGVHD) were lower (20.0% vs 51.9%) (P = 0.026) (5.0% vs 33.3%) (P = 0.040), the incidence of CMV infection after transplantation was higher (81.8% vs 51.9%) (P = 0.028). In addition to these, the differences were not statistically significant in term of implantation level, PLT reconstitution, aGVHD, lung infection, hemorrhagic cystitis, 1-year relapse and survival between the 2 groups (P0.05).The transplantation of URD-PBSC combined with UC-MSC is effective and safe. The speed of neutrophils reconstitution is faster. The incidence and severity of cGVHD are lower, but the attention should be paid to prevent the CMV infection.

This study was aimed to explore the new mechanism of α-Galactosyleramide (α-GalCer), a synthetic glycolipid, and a well-known activator of natural killer T cells (NKT) for improving acute graft-versus-host disease(aGVHD).Murine allogeneic bone marrow transplantation (allo-BMT) model was established. Recipient mice were injected intraperitoneally with α-GalCer immediately after allo-BMT, whereas mice from the vehicle groups received the diluent (DMSO) only. The severity degree of aGVHD was estimated by survival, aGVHD clinical score and pathology. The mechanism of aGVHD reduced by α-GalCer was explored by detecting T cells migration in vivo and in vitro.Mice in α-GalCer group survived longer than in control group, and their clinical and pathological status of aGVHD were lighter. α-GalCer reduced aGVHD by altering donors T cell migration.After allo-BMT α-GalCer reduces aGVHD by altering donor T cells migration.

This study was purposed to investigate the effect of umbilical cord mesenchymal cells (UC-MSC) infusion on the pulmonary infection in haploidentical hematopoietic stem cell transplantation (hi-HSCT). The infection of 83 patients underwent hi-HSCT was detected and analysed, among them 42 patients received haploidentical hi-HSCT, 41 received hi-HSCT combined with UC-MSC infusion. The results showed that 31 cases (73.81% ± 6.78%) were infected by cytomegalovirus and 21 cases in patients received hi-HSCT experienced pulmonary infections, including infections of fungal, virus, bacteria, tubercle bacillus, PCP and so on, the incidence rate was (50 ± 7.72)%; the infection of cytomegalovirus (CMV) was found in 31 cases, the incidence rate was (78.05 ± 6.46)%. In patients received hi-HSCT combined with UC-MSC, only 15 patients experienced pulmonary infection, the incidence rate was (36.59 ± 7.52)%, and the infection of cytomegalovirus (CMV) was observed in 32 patients, the incidence rate was (78.05 ± 6.46)%. There was no obvious statistical difference between two groups(P0.05). It is concluded that the UC-MSC infusion not increases the infection rate in hi-HSCT.

This study was aimed to investigate the effect of dendritic cell-derived exosome (DCex) on in vitro osteoblast differentiation of human bone marrow mesenchymal stem cells (hBM-MSC). DCex were harvested from the DC culture supernatants by ultracentrifugation. The morphology of DCex was observed by using transmission electron microscopy and the surface marker expression was detected by flow cytometry. MSCs at passage 3 were used in this study. DCex incorporation into MSCs was observed under a confocal microscope. MSCs were either exposed to DCex (10 µg/ml) or the standard osteogenic induction condition. The cells cultured in complete medium were served as the control. The expression levels of Runt related transcription factor 2 (Runx2) were detected by real-time and standard PCR. The cellular alkaline phosphatase (ALP) activity was also detected. The results showed that the DCex were spherical or oval membrane vesicles with diameters of about 40-100 nm under transmission electron microscope. The DCex expressed surface molecules specific for DCs, including CD83, CD86, CD80, and HLA-DR. After cultured for 7 days, the MSCs treated with DCex highly expressed Runx2 as compared with the control group (P0.05). After cultured for 14 days, ALP activity of the DCex-treated MSCs was markedly higher than the control group (P0.01), though it was lower than that of MSCs treated with standard inductive agents. It is concluded that DCex can induce MSCs to differentiate into osteoblasts. The detailed investigations are needed to clarify the underlying mechanisms.

This study was purposed to investigate the immune state of the patients suffered from pulmonary infection within 6 months after haploidentical hematopoietic stem cell transplantation (hi-HSCT). Adenosine triphosphate (ATP) value in CD4(+) T cells was measured by ImmuKnow method to assess the function of the lymphocytes in peripheral blood of 25 patients at 6 months after hi-HSCT. The results showed that the ATP level in CD4(+) T cells of the patients suffered from pulmonary infection was (179.88 ± 65.41) ng/ml before transplantation, (172.69 ± 118.81) ng/ml at 1 month, (218.15 ± 124.26) ng/ml at 3 months, (313.42 ± 116.29) ng/ml at 6 months after transplantation. The ATP level in CD4(+) T cells of the patients without pulmonary infection was (210.44 ± 94.71) ng/ml before transplantation, and decreased to (193.66 ± 133.69) ng/ml at 1 month and increased gradually to (355.02 ± 43.38) ng/ml at 3 months, (355.73 ± 93.85) ng/ml at 6 months after transplantation. It is concluded that the low ATP value in CD4(+) T cells in patients prior and post hi-HSCT may suggest probability of occurrence for infections, ATP value in CD4(+) T cells may be used as a reference indicator for clinical empirical use of antibiotics.

The aim of this study was to investigate the reconstitution of CD4(+)CD25(+) T cells after haplo-identical bone marrow transplantation (hiBMT) and its correlation with graft versus host disease (GVHD) and relapse. Peripheral blood samples from 27 patients after hiBMT were harvested and the percentage and absolute counts of CD4(+)CD25(+) T cells were detected by flow cytometry. The correlations of GVHD occurrence and disease relapse with the reconstitution of CD4(+)CD25(+) T cells were analyzed. The results showed that the percentage of CD4(+)CD25(+) T cells of peripheral blood samples increased significantly after G-CSF priming. At day 30 after hiBMT, CD4(+)CD25(+) T cells were recovered to the 20% of normal level, followed by a slowly process in 3 months, and up to one half of the normal level at 180 days. There was no evidence to prove relationship between CD4(+)CD25(+) T cells and acute GVHD, while CD4(+)CD25(+) T cells were increased significantly in the chronic GVHD group. The absolute count of CD4(+)CD25(+) T cells showed no relations with relapse of leukemia during the first year after hiBMT. In conclusions, chronic but not acute GVHD was in relation to the reconstitution of CD4(+)CD25(+) T cells based on the anti-CD25 antibody therapy model for the prevention of GVHD after hiBMT. Further investigation is needed to clarify whether the relapse of leukemia after hiBMT is related to the reconstitution of CD4(+)CD25(+) T cells.

This study was aimed to explore the effect of donor characteristics (age, sex and so on.) on CD34(+) cell yields in apheresis from healthy donors mobilized by recombinant granulocyte colony-stimulating factor(rhG-CSF). In 61 healthy donors, the characteristics associated with CD34(+) cell yield were analysed. The relationship between the CD34(+) cell yields and donor characteristics was statistically assessed with multivariate forward, backward and stepwise regression methods. A variety of parameters were analyzed which included donor age, sex, weight, height, body mass index (BMI) and time for collection of peripheral blood apheresis, while the mean number of peripheral blood mononuclear cells (MNCs), CD34(+) cell count, CD34(+) cell proportion based on MNC and CD34(+) cell count per kg of donor weight were used as the variables. The results showed that age of donors was the main factor impacting CD34(+) cell yields (-0.60r-0.45, p0.005). In a partial correlation analysis the body weight, height and BMI were served as control factors, the negative correlation of age with CD34(+) cell yields was still found (-0.50r-0.35, p0.02). BMI was only weakly correlated with the yields of CD34(+) cells per kg(r = -0.297, p0.05). As a whole, sex showed no relation with the CD34(+) cell yields. Compared with the female group less than 35 years old, height, weight and BMI in male group of low age exerted a positive impact on CD34(+) cell yield. The optimal time for collection of PB was day 4 after treatment with rhG-CSF, when 70% of the donors could reach the peak CD34(+) cell yields. It is concluded that the age of the donors is the first factor determining the choice of donors for allogeneic hematopoietic stem cell transplantation, the sex, height, weight and BMI are secondary factors impacting yield of CD34(+) cells from donors mobilized with rhG-CSF.

In order to explore the diagnosis and therapeutic effectiveness of nocardiosis after allogenic hematopoietic stem cell transplantation (allo-HSCT), the features of clinical manifestation, laboratory examination and response to TMP-SMZ treatment in two cases of nocardiosis after allo-HSCT were analyzed retrospectively. The result showed that the attack happened to 2 patients at day 15 and 170 after allo-HSCT respectively, displaying fever and chest pain. Chest CT scan indicated bilateral pulmonary tuberculous shadow. Nocardiosis was diagnosed by the culture of sputum, bronchoalveolar lavage (BAL) fluid and pus samples as well. Both of these cases exhibited good response to combined therapy containing TMP-SMZ for half a year. It is concluded that nocardiosis is a rare complication after allo-HSCT, in which pulmonary involvement is commonly observed. The culture of BAL fluid is helpful for its diagnosis and this disease might be sensitive to the treatment of TMP-SMZ-containing regimens.

The purpose of this study was to investigate the feasibility and clinical outcome of granulocyte colony stimulating factor (G-CSF)-mobilized haploidentical bone marrow transplantation combined with peripheral blood stem cells (hiBM+PBSCT) for therapy of leukemia. 125 leukemia patients underwent G-CSF primed haploidentical stem cell transplantation without ex-vivo T cell depletion. All haploidentical donors were injected with G-CSF at dose of 5 microg/(kg.d) for 7 days. The patients were divided into groups A and B. 29 patients in group A underwent hiBM+PBSCT at 7th and 8th days of mobilization in donors with G-CSF respectively; 96 patients in group B underwent hiBMT. All patients received the same GVHD prophylaxis regimen, the clinical outcomes were investigated. The results showed that all patients except one CML-myelofibrosis patient achieved trilineage engraftment. Engraftment median times were 15 and 19 days for neutrophil and platelet in group A respectively, while engraftment median times were 18 and 23 days for neutrophil and platelet in group B respectively. The incidences of grade II-IV aGVHD were 31.03% in group A and 12.5% in group B respectively (p0.05). The incidences of grade III-IV aGVHD was 13.79% and 10.41% in group A and group B (p0.05). The aGVHD-related death incidence was 3.45% and 5.21% in group A and group B (p0.05). The incidence of grade II-IV cGVHD was 48.2% and 35.4% in group A and group B respectively (p0.05). The incidence of extensive cGVHD was 23.3% and 15.6% in group A and group B respectively (p0.05). The disease relapse rate was 6.8% (2/29) and 18.75% (18/96) in group A and group B respectively (p0.05). It is concluded that the G-CSF-mobilized allogeneic haploidentical BM plus peripheral blood HSCT without T cell depletion provides a rapid and sustained engraftment without increase of severe GVHD, furthermore, the relapse rate of disease is reduced remarkably, thus this method can be used in clinic.

This study was purposed to explore the efficacy of hematopoietic reconstitution and survival of patients with myelodysplastic syndrome (MDS) after allogeneic hematopoietic stem cell transplantation (allo-HSCT). Allo-HSCT without T lymphocyte depletion was used in 6 patients with MDS from November 1999 to June 2007. 4 cases out of them received allo-PBSCT from HLA matched sibling donors with conditioning regimen of cyclophosphamide (CTX) and Bu. Graft versus host disease (GVHD) was prevented by the administration of immunosuppressive drugs of cyclosporine A (CsA) and short-course MTX. 2 patients received haploidentical allogeneic bone marrow transplantation (hi-alloBMT) after preconditioning with cytosine arabinoside (Ara-C), CTX and total body irradiation (TBI) with a linear accelerator. GVHD was prevented by the administration of immunosuppressive drugs including CSA, short-course MTX, MMF, anti-CD25 monoclonal antibody and ATG. The results showed that all of the patients were engrafted successfully. The median time of granulocyte recovery exceeding 0.5 x 10(9)/L and platelets exceeding 20 x 10(9)/L were days 15 and 20.3 respectively, and 100% donor hematological cells were detected by cytogenetic analysis. All patients did not experience serious acute graft-versus-host disease (aGVHD). During 18 - 108 months of following-up, 2 cases died of pulmonary complication and of relapse; the other 4 cases survive in a disease-free situation. In conclusion, allo-HSCT was an effective approach for the treatment of MDS.

This study was aimed to detect the changes of bcr/able gene level in ph+ CML patients at different stages after allo-HSCT by real-time quantitative PCR and to evaluate the significance of this detection. The serial detection of bcr/abl fusion gene levels in 21 cases of CML treated with allo-HSCT was performed by RQ-PCR. The results showed that the bcr/able fusion gene could not be detected in 7 out 21 CML cases with positive fusion gene after allo-HSCT, while the bcr/abl fusion gene of different levels could be detected in 14 cases within 1-6 months. Dynamic detection indicated that the bcr/abl fusion gene levels in 9 cases were lower with relative value 0.0074%-0.088% and then could not be detected within 3-7 months after allo-HSCT. The bcr/abl fusion gene levels in 5 cases diagnosed as molecular relapse were between 0.077%-75%. The bcr/abl fusion gene levels in 1 out of 5 cases were 0.95%, 1.5%, and 0.16% in month 1, 2 and 3, respectively, and turned to negative in the month 4 without any treatment after allo-HSCT. 2 cases received the donor peripheral blood stem cell infusion, and then their bcr/abl mRNA levels could not be detected in bone marrow. Another 2 cases developed to the hematologic relapse, 1 out of 2 cases reached CR again after infusion of donor peripheral blood stem cells and chemotherapy, the other one died. It is concluded that serial quantifications of bcr/abl mRNA levels by RQ-PCR are reliable and can be used to detect the MRD, to monitor the outcome and to predict the relapse.

This study was purposed to investigate the correlation between the dose infused megakaryocytic precursors (CD34+, CD34+CD61+) and recovery time of platelet count following an allogeneic PBSCT and/or BMT through quantitative detection of CD34+ and its subpopulation in peripheral blood and BM mobilized by G-CSF. 24 patients with various hematologic malignancies received PBSCT/BMT from their HLA matched or unrelated donors and haploidentical siblings in April-December 2007. 20 evaluated patients were divided into 2 groups according to different transplant schemes. HLA matched group received PBSCT regime and haploidentical group received PBSCT combined with BMT. CD34+CD61+ subpopulations in sample from patients receiving PBSCT/BMT were measured by flow cytometry immediately or storage over night. The results showed that the median number of infused CD34+, CD34+CD61+ and CD34-CD61+ cells in haploidentical group were 6.24x10(6)/kg (1.53-20.48), 66.19x10(4)/kg (8.16-493.83), and 34.38x10(6)/kg (14.71-109.16) respectively, in HLA matched group those were 4.88x10(6)/kg (1.00-8.24), 14.16x10(4)/kg (11.63-96.87), and 13.50x10(6)/kg (1.74-35.61), respectively. Median days of ANCs0.5x10(9)/L and platelets20x10(9)/L were 18.5 (11.0-29.0) days and 16.5 (9.0-35.0) days in haploidentical group respectively; in HLA matched group those were 14.5 (9.0-24.0) and 10.5 (6.0-37.0) respectively. A significance difference of median days for ANC engraftment presented between two groups (p=0.048). There was no significant difference of time for platelet engraftment between 2 groups. For patients with CD34+ cell dose2x10(6)/kg there was significant difference of time of platelet engraftment between HLA matched and haploidentical groups (p=0.006). The number of CD34+CD61+ cells infused in 12 haploidentical patients or in 8 HLA matched patients were much better correlated with the time of platelet recovery up to 20x10(9)/L than that of number of CD34+ cells infused in total 20 patients (r=-0.768 and p=0.004 for haploidentical CD34+CD61+ cells, r=-0.747 and p=0.033 for HLA matched CD34+ CD61+ cells, r=-0.449 and p=0.047 for CD34+ cells). There was an inverse correlation between the number of infused CD34+ CD61+ cells and time of platelet engraftment. Therefore, as the number of CD34+ CD61+ cells increased, duration of platelet engraftment (time to reach platelet count of 20x10(9)/L) shortened significantly. It is concluded that the determining the number of megakaryocytic precursor by flow cytometry may predict the platelet reconstitutive capacity of the allogeneic hematopoietic stem cell transplantation, which is in haploidentical PBSCT and in BMT.

In order to evaluate the diagnostic value of fibrotic bronchoscopy (FB) in the pulmonary infiltration following bone marrow transplantation (BMT), 18 patients with pulmonary complications after BMT from November 2003 to March 2006 were performed with FB. Bronchoalveolar lavage (BAL) and brushing were performed in patients who had received short-term empirical therapy without good response, and transbronchial lung biopsy (TBLB) was carried out in 3 cases. The results showed that 9 out of 10 cases with pulmonary infection, including bacterial pneumonia (n = 3), aspergillosis (n = 2), pneumocystis carinii pneumonia (n = 3) and viral infection (n = 1) were diagnosed by using FB. One case was diagnosed as tuberculosis after open lung biopsy following negative results from twice BAL. 2 out of 8 cases were diagnosed by TBLB as noninfectious pulmonary complications. In conclusion, FB, especially with BAL, is a safe and useful procedure for the evaluation of pulmonary complications, which is particularly suitable for diagnosis of pulmonary infection after BMT. Furthermore, TBLB should be recommended in order to avoid open lung biopsy, if the patients tolerate the operation.

The purpose of study was to investigate the impact of killer immunoglobulin-like receptor (KIR) and its ligand on haploidentical bone marrow transplantation. 74 cases were analyzed for the distribution frequencies and characteristics of KIR and its ligand as well as the impact of KIR ligand for the haploidentical bone marrow transplantation in terms of the overall survival, disease-free survival (DFS), GVHD and relapse. The results showed that among the 19 KIR genotypes currently nominated KIR2DL1, KIR2DL4 and KIR3DL2-3 could be detected in all the cases. Other high frequency genotypes included KIR3DP1 (98.6%), KIR2DP1 (98.6%), KIR3DL1 (97.3%) and KIR2DL3 (97.3%). Inhibitory receptor genotypes were 1.37-fold of activating receptor genotypes. KIR2DL1, KIR3DL2, KIR3DL3 and KIR2DL4 were found in all haplotypes and at least one genotype of KIR2DL2 and/or KIR2DL3 existed in all haplotypes. Among the 14 genotypes found in the test, the HLA-Cw7 was the most popular (37.8%) and the group 2 (HLA-Cw1, 3, 7, 8, 13, 14) recognized by KIR2DL2/2DL3 counted for 43.2%. The incompatibility of KIR for 32 cases of haploidentical BMT was 43.8%, of which 9/14 were KIR2DL incompatible, 5/14 were KIR2DL2 or KIR3DL1 incompatible. Among the 46 cases of haploidentical BMT, 29 cases were HLA-Cw matched and 14 cases were mismatched. The completed mismatch ratio of HLA-Cw was 30.4% and the match ratio was 63.4%. The survival rate was higher for the 14 cases of KIR genotype compatible group than the 13 cases of KIR genotype incompatible group (p = 0.032). The disease-free survival was significantly higher for the 17 cases of mismatched KIR ligands (HLA-Cw) group than the matched group (p = 0.024). The survival rate was higher in GVHD group than that in non-GVHD group when the KIR ligand was missing. The acute and severe GVHD was related to the existence of activating receptor of KIR2DS1/2DS2. The incompatibility group was accompanied with frequent acute and severe GVHD and less relapse and vice versa for the compatibility group. One patient died after BMT among the 14 mismatched KIR ligand group suffering from myelogenous leukemia while 4 patients out of 12 patients died in the matched group. It is concluded that the haploidentical BMT is characterized by mismatch between donor and recipient and its immunological reactions also features by the incompatibility of KIR genotype and missing ligand. The missing ligand for the donor KIR has strong effect on the outcome of BMT and it means a lot to analyze the KIR genotype and its ligand for the selection of best donor and prognostic evaluation in haploidentical BMT.

To investigate the leukemia relapse of AL patients after HLA haploidentical bone marrow transplantation (HLA HBMT), 2 relapsed leukemia patients received HLA HBMT were studied, peripheral blood simples and bone marrow smear were examined, morphologic change of bone marrow cells was observed, while the HLA genotype and chromosome karyotye were analyzed by PCR and routine G-banding methods, respectively. The results indicated that the two cases were diagnosed primarily as acute lymphocytic leukemia (common cell subtype) and acute megakaryocytic leukemia, in which chromosome abnormalities or activation of protooncogene in leukemic cells were observed. The complete hematopuietie reconstitution of donor origin was obtained in these 2 cases after HLA HBMT, but the leukemic cells in these 2 leukemia patients were confirmed to be donor origin after relapse, their blood groups and HLA genotype were found to be originated from donor. These 2 relapsed leukemia patients were diagnosed as acute lymphocytic leukemia (B cell subtype) and acute megakaryocytic leukemia. It is suggested that high-dose of immunosuppressive agents used in transplantation may contribute to leukemia relapse of donor origin in these patients. Abnormalities in hematopoietic microenvironment may be also involved in the leukemia development. Donor-cell leukemia after allogeneic hematopoietic stem cell transplantation can be an ideal model to investigate the related events in human leukemogenesis.

To explore the occurrence patterns of neurological complications following haploidentical bone marrow transplantation, a series of clinical data as the onset time, etiology, choices of therapies and prognosis in 10 patients with neurological disorders were summarized. The results showed that complications occurred in 10 out of 74 patients after bone marrow transplantation, which include 3 with encephalorrhagia, 3 infection, 1 epilepsy, 1 Guillian-Barr's syndrome, 1 encephalopathy and 1 schizophrenia. 4 patients died of neurological complications.neurological complications commonly occurred in haploidentical bone marrow transplantation, and encephalorrhagia might be the main indication that needs intensive care. Moreover, central nerve system infection and peripheral nerve diseases associated with slow immune reconstitution should have clinical interests.

To explore the occurrence patterns of pulmonary complications at different stages in haploidentical bone marrow transplantation, a series of clinical data as the onset time, etiology, management choices and prognosis in 18 patients with pulmonary disorders were summarized. The results showed that in 18 out of 70 patients after bone marrow transplantation occurred pulmonary complications which included pneumonia affected by bacteria (7 cases), fungus (5 cases) and cytomegalovirus (CMV, 4 cases), bronchiolitis obliterans organizing pneumonia (BOOP, 1 case), and idiopathic pneumonia syndrome (1 case), out of which 8 cases died. Fungal and CMV pneumonia occurred predominantly 2 to 3 months after transplantation, whereas bacterial pneumonia was observed in the duration of 3 to 12 months and 4 cases of them suffered from secondary fungal infections during treatment. BOOP and idiopathic pneumonia syndrome were diagnosed 12 months and 50 days after transplantation respectively.pulmonary complications were commonly seen in haploidentcal bone marrow transplantation, and fungal pneumonia might be the main cause that needs intensive management.

To investigate the properties of haploidentical donor-derived bone marrow engraftment and hematopoietic reconstitution in patients received bone marrow transplantation, 15 patients with leukemia received bone marrow grafts without T cell depletion from their family donors of those with 2-3 mismatched loci of HLA antigens. The donors were given G-CSF 250 micro g/day for 7 days prior to marrow harvest. All patients were treated with conditioning regimens consisting of high-dose of Ara-C, cyclophosphamide, and total body irradiation. A four-agent based GVHD prophylaxis was used as cyclosporine A, MTX, ATG and mycophenolate mofetile (MMF). Donor engraftment was evaluated as identification of HLA locus, chromosome karyotype, DNA fingerprinting, blood type and other parameters such as occurrence of GVHD, recovery of peripheral blood cell counts as well as normal myelogram. The results showed that successful and stable engraftment was established in all patients. The median time of granulocyte counts0.5 x 10(9)/L and platelet20 x 10(9)/L was 18 (13-23) and 22 (16-32) days, respectively. One of the patients relapsed despite the bone marrow chimerism appearing after transplantation. The grade I acute GVHD occurred in 8 and grade II-IV in 5 of the 15 patients. Of the patients, 7 received marrow grafts from donors of opposite sex were identified for donor engraftment by chromosome analysis, 4 by blood typing, 7 with HLA locus analysis and 1 with DNA fingerprinting. In conclusion, HLA haploidentical bone marrow transplantation is feasible with a series of management including mobilization with G-CSF in donors, intensive conditioning and proper immunosuppressants, which enable the allo-transplants to stride across the immunological barrier.

To explore the antitumor activities of KRN7000 in a NS-1 myeloma-bearing mice and the underlying mechanisms, cytotoxic activities of the spleen cells treated with KRN7000 in vivo and in vitro were examined with Yac-1 (a NK-sensitive cell line) and NS-1 (a NK-insensitive cell line) as the targeted cells. The life span of NS-1 myeloma-bearing mice was observed after treating with KRN7000 and combined with cyclophosphamide (CTX). Furthermore, toxicities of KRN7000 administration on liver and kidney were evaluated. The results showed that KRN7000 enhanced NK cytotoxic activities of spleen cells. Around 20% of NS-1-bearing mice survived after KRN7000 administration and the survival rate reached up to 80% of NS-1-bearing mice when KRN7000 was used in combination with CTX at a dose of 100 mg/kg (P0.005). KRN7000, at a dose of 100 micro g/kg, had no toxic effects on liver and kidney. These findings suggest that KRN7000 might be a promosing agent in tumor immunotherapy.

Although chemotherapy can achieve a high rate of disease remission in patients with newly diagnosed acute leukemia, patients with recurrent or refractory leukemia generally have a poorer rate of response. This study was designed to assess the utility of high-dose mitoxantrone and intermediate-dose cytarabine in the treatment of patients with relapsed or refractory acute leukemia. Thirty patients with relapsed or refractory acute leukemia were treated with mitoxantrone at a total dose of 40 mg/m(2) intravenously and cytarabine 1 - 1.5 g/m(2) over 3 hours once daily for five doses. Twenty-six of thirty patients achieved complete response (CR 87%) and one achieved partial response (PR 3.3%). No patient died during the induction course. The main toxic effect was hematopoietic suppression that was clinically acceptable. The median time needed for CR was around 30 days. The median disease-free and overall survival times were 3.5 and 6 months, respectively. The results demonstrate that short course high-dose mitoxantrone with cytarabine is associated with a trend toward a higher CR rate and therefore, it should be an effective antileukemic regimen for the treatment of refractory and relapsed acute leukemia.

To explore the feasibility of using CD25 monoclonal antibody (Basiliximab) in T-cell undepleted allo-BMT with graft from haplotype-matched related donor for acute GVHD prophylaxis. Twenty-eight patients with leukemia received allo-BMT from HLA two or three loci mismatched related donors. The donors were given G-CSF (Lenograstim) 250 micro g/d for 7 doses prior to marrow harvest. CSA, MTX, ATG and mycophenolate mofetil (MMF) were combined for GVHD prophylaxis. ATG 5 mg/(kg.d) was infused from day 4 to day 1 before transplantation and MMF was administered from day 7. In the study group, the patients received additional CD25 monoclonal antibody for aGVHD prophylaxis. CD25 20 mg each by 30 min intravenous infusion on 2 hours before transplantation and day 4 after transplantation was administered while no application of CD25 in the controls. The outcomes of transplantation were compared between the stud y and control groups. The results showed that the median number of CD34(+) cell in graft was 5.9 x 10(6)/kg in the control group and 7.9 x 10(6)/kg in the study group. The median number of CD3(+) cell was 48 x 10(6)/kg and 52 x 10(6)/kg respectively (P0.05). All patients showed 100% donor-typed hematopoietic cells after transplantation by cytogenetic evidence. Five out of fifteen patients in the control group experienced II - IV acute GVHD. While none of thirteen in the study group developed the II - IV acute GVHD. However, none in both groups developed extensive cGVHD. The median follow-up duration was 8 (3-15) months in the study group and 26 (15-36) months in control. In the study group, one patient died from transplant related mortality (CMV infection); no one relapsed; and 12/13 patients survived in disease-free situation within the period of follow-up. In the control group, six patients died from transplant related mortality (3 GVHD, 2 infection and 1 relapsed) and 9/15 patients survived in disease-free situation. The one-year probabilities of disease-free survival (DFS) in two groups were significantly different (P0.05). It is concluded that the transplant from haploidentical donors used CD25 antibody is effective and feasible in preventing acute severe GVHD and improving DFS. The major histocompatibility barrier in the haploidentical related allo-BMT could be crossed by donors primed with G-CSF and GVHD prophylaxis with CD25 antibody in the hosts.

To observe normal bone marrow stromal function for supporting hematopoiesis and realize the correlation of bone marrow stromal function in patients with aplastic anemia (AA) and anisodamine therapy, normal human marrow CFU-GM was assayed on bone marrow stromal layer (SL) formed on week 3 culture, and the CFU-GM without SL as a control (100%). Results showed that the production of CFU-GM on the normal SL was significantly higher than that of the control. The production of CFU-GM on the SL of 4 AA patients, who responded to anisodamine, was80% of the control, and when 2 of them were reexamined for stromal function after treatment, the number of CFU-GM on SL rose up to 168.8% and 249.2% from 38.7% and 39.7% of the control respectively. While in the rest 6 patients, the number of CFU-GM was80% of the control, only one patient was improved by anisodamine therapy. So, normal bone marrow stromal layer can obviously support the growth of granulocyte/macrophage progenitor cells. Anisodamine therapy could be an effective agents for aplastic anemia with stromal dysfunction