目的 研究急性白血病KG1a细胞抗原体外诱导人外周血单个核细胞(peripheral blood mononuclear cells,PBMC)的杀伤活性和T细胞受体Vαβ谱系的表达和克隆情况.方法 应用制备的急性白血病细胞KG1a抗原和外周血单个核细胞混合培养,LDH释放法检测诱导前后PBMC杀伤KG1a细胞活性,RT-PCR扩增诱导前后T细胞的TCR Vαβ亚家族的CDR3,并用基因扫描分析T细胞受体Vαβ谱系的表达情况.结果 经KG1a细胞抗原诱导后PBMC细胞杀伤活性增强,在不同效靶比时诱导前后的杀伤率差异有统计学意义(P<0.05),且随效靶比增加而增加;CD8+T淋巴细胞比例显著提高(P<0.05).诱导后T细胞出现单克隆趋势、单克隆及寡克隆,某些亚家族甚至消失.结论 KG1a细胞抗原体外可诱导T细胞呈克隆性增殖,具有特异性细胞毒作用,能提高对KG1a细胞的杀伤活性.

目的 探讨CD34+ CD38-KG1a白血病干细胞抵抗同种异体自然杀伤(NK)细胞杀伤作用的机制.方法 采用免疫磁珠法(MACS)分选CD34+ CD38-KG1a细胞和4例健康个体外周血中NK细胞,并采用流式细胞术检测纯度;乳酸脱氢酶(LDH)释放法检测NK细胞在不同效靶比(5∶1,10∶1,20∶1)下对K562和CD34+ CD38-KG1a细胞的杀伤作用;聚合酶链式反应-序列特异性引物法分析NK细胞KIR和CD34+ CD38-KG1a细胞HLA-Ⅰ基因分型;流式细胞术检测K562和CD34+ CD38-KG1 a细胞表面主要组织相容性复合体(MHC)-Ⅰ类链相关分子A/B、人巨细胞病毒UL16结合蛋白(ULBP)1～3和人类白细胞抗原(HLA)-Ⅰ类分子的表达.结果 分选的CD34+ CD38--KG1a细胞和NK细胞纯度分别为(94.25±2.16)％、(91.70±2.05)％.NK细胞在各效靶比下对CD34+ CD38-KG1a细胞的杀伤率均低于K562细胞(P＜0.05).4例健康个体NK细胞KIR基因型为KIR2DL1、KIR2DL3、KIR3DL1和KIR3DL2,CD34+ CD38-KG1a细胞HLA-Ⅰ基因型为A30、30,B51、78,Cw4、16.K562细胞高表达NKG2D配体,CD34+CD38-KG1a细胞几乎不表达NKG2D配体,CD34+ CD38-KG1a细胞中NKG2D配体表达率显著低于K562细胞(P＜0.05);K562细胞中几乎不表达HLA-Ⅰ,CD34+ CD38-KG1a细胞高表达HLA-Ⅰ,CD34+ CD38-KG1a细胞中HLA-Ⅰ的表达率显著高于K562细胞(P＜0.05).结论 CD34+ CD38-KG1a细胞明显抵抗同种异体NK细胞的杀伤作用,其机制可能与高表达HLA-Ⅰ分子和低表达NK2D配体有关.

目的 研究中药砷复合制剂青黄散(由青黛和雄黄二药组成)组分靛玉红(Indirubin,青黛的有效成分)和二硫化二砷(Arsenic Disulfide,As2S2,雄黄的有效成分)对非M3型急性髓系白血病(AML)KG1a细胞株的单独和联合杀伤作用,并探讨相关杀伤作用机制.方法 采用瑞氏姬姆沙染色法观察靛玉红与As2S2作用于KG1a细胞的形态,采用流式细胞术检测靛玉红与As2S2作用于KG1a细胞凋亡率,采用CCK-8检测靛玉红与As2S2作用于KG1a细胞抑制率,进一步用Western blot检测KG1a细胞Bcl-2、Smac、XIAP-1、c-IAP、Caspase-3蛋白的表达.结果 青黄散组分靛玉红与As2S2单独对KG1a细胞均具有增殖抑制作用,联合用药的增殖抑制率、凋亡率作用更明显(P＜0.05),CompuSyn软件分析靛玉红和As2S2以1:1组合对KG1a细胞株的联合指数(CI)＜0.9.联合用药在形态学上能明显促进细胞的凋亡变化,增加细胞的Caspase-3和Smac蛋白表达,抑制BCL-2、XIAP-1、c-IAP蛋白的表达.结论 青黄散组分靛玉红与As2S2能联合抑制KG1a细胞增殖和诱导其凋亡,其机制与抑制BCL-2、XIAP-1、c-IAP蛋白表达,促进Caspase-3、Smac蛋白表达相关.本药可用于治疗常规药物抵抗的AML并为非M3型AML的治疗提供相关实验依据.

Background Radioresistance is a major challenge during the treatment of NK/T cell lymphoma. This study aimed to investigate the potential role of MicroRNA-150 (miR-150) in increase the sensitivities of NK/T cell lymphoma to ionizing radiation. Results In this study, we found that miR-150 was significantly decreased in NK/T cell lymphoma tissues and cell lines. Low expression of miR-150 was positively associated with therapeutic resistance in 36 NK/T cell lymphoma cases. Our further in vitro and in vivo studies illustrated that overexpression of miR-150 substantially enhanced the sensitivity of NK/T cell lymphoma cells to ionizing radiation treatment. Furthermore, luciferase reporter assays in NK/T cell lymphoma cells transfected with the AKT2 or AKT3 three prime untranslated region reporter constructs established AKT2 and AKT3 as direct targets of miR-150. The phosphatidylinositol 3-kinase inhibitor LY294002 was used to inhibit Akt to verify miR-150 increase NK/T cell lymphoma cell radiorsensitivity through suppress the PI3K/AKT/mTOR pathway. Conclusions Taken together, this study demonstrates that miR-150 might serve as a potential therapeutic sensitizer through inhibition of the AKT pathway in NK/T cell lymphoma treatment.

目的:比较每日1次和每日4次静脉滴注含白消安预处理方案行异基因造血干细胞移植(Allo-HSCT)的疗效和安全性.方法:回顾性应用白消安方案预处理行Allo-HSCT且生存期超过3个月的恶性血液病患者72例,根据给药方式分为每日1次静脉滴注组(43例)和每日4次静脉滴注组(29例),比较2组患者的3年累计生存率评估其有效性;比较2组患者的造血重建情况、移植相关并发症、移植物抗宿主病和复发情况评估其安全性.结果:2组患者的3年累计生存率差异无统计学意义;2组患者的造血重建、移植物抗宿主病发生率、复发率差异无统计学意义;2组患者的移植相关并发症比较,白消安每日1次静脉滴注组3～4级黏膜炎的发生率更高,差异有统计学意义(P＜0.05),2组患者其他并发症差异无统计学意义.结论:恶性血液病的Allo-HSCT,预处理方案采用白消安每日1次静脉滴注的给药方案同每日4次给药方案具有相似疗效,但前者3～4级黏膜炎的发生率可能更高.

OBJECTIVE:To explore the effects of blocking TCR-CD3 and B7-CD28 signals on immune function of mice with chronic GVHD by using TJU103 and CTLA4-Ig.METHODS:On the basis of foregoing murine model of chronic GVHD, according to interference modes after infusion 6×107 spleen cells of donor mice, the recipients were divided into 5 groups: blank control, cGVHD, TJU103 interference, CTLA4-Ig interference and TJU103+CTLA4-Ig interference groups. The score of clinical manifestation and tissue histopathology were used to evaluate the effects of all the interferences on chronic GVHD.RESULTS:TJU103 and CTLA4-Ig could not influence the formation of the mouse chimera. The analysis of Kaplan survival curve of mice with chronic GVHD showed that the CTLA4-Ig and TJU103+CTLA4-Ig reduced the incidence of chronic GVHD, the TJU103 could delay the occurrence of chronic GVHD, but all the interference factors could not change the severity of chronic GVHD.CONCLUSION:TJU103 can delay the onset time of chronic GVHD, and the CTLA4-Ig can reduce the incidences of cGVHD, the combining use of TJU103 and CTLA4-Ig can significantly reduce the incidence of chronic GVHD, but can not change the severity of chronic GVHD.

Objective:To investigate the expression of MicroRNA-150 (miR-150) in NK/T cell lymphoma tissues and its effect on radio-sensitivity of NK-92 cells.Methods:Thirty-six patients with NK/T cell lynphoma that treated in Zhujiang Hospital Affiliated to Southern Medical University were included as study subjects,and their tissue samples were collected.All the patients received similar radiotherapy,and the short-term efficacy was evaluated by the standard of International Workgroup (IWG) of Lymphoma.The patients were further divided into CR (complete remission) group and Non-CR (non-complete remission) group according the treatment efficacy.The miR-150 expression in lymphoma tissues and NK/T cell lymphoma cell lines were detected by qRT-PCR.The NK-92 cells were then transfected with miR-150 mimics.The effect of miR-150 mimics on NK-92 cell radiosensitivity was analyzed by MTT and colony formation assay;effect of miR-150 mimics on radiation induced apoptosis of NK-92 cells were analyzed by Flow Cytometry;and the effect of miR-150 mimics on the expression of apoptosis-related proteins (Caspase 3 and PARP) in NK-92 cells was determined by Western blotting.Results:According to IWG criteria,12 patients had CR while the other 24 patients had Non-CR.Compared with CR group,the microRNA-150 level in NonCR group was significantly decreased(P＜0.05).Compared with the normal sCD3-CD56+ NK cells,miR-150 was significantly lower in NK/T cell lymphoma tissues (9.10 ±0.19 vs 4.01± 0.22 P＜0.01) and five NK/T cell lymphoma cell lines (P＜0.05);over-expression of miR-150 significantly decreased the proliferation and colony formation of NK-92 cells (all P＜0.01),and increased the sensitivity enhancement ratio (SER) after radiation (the SER was 5.375 after 10 Gy exposure;in addition,over-expressions of miR-150 significantly promoted the radiation-induced apoptosis of NK-92 cells [(37.3± 1.24)％ vs (28.3±2.34)％,P＜0.05],and promoted the protein expression of caspase 3 and PARP in NK-92 cells.Conclusion:miR-150 expression significantly decreased in both NK/T cell lymphoms tissues and cell lines.The patients of low miR-150 expression had low CR after radiotherapy.miR-150 mimics transfection promoted radiation-induced apoptosis of NK-92 cells.miR-150 has an enhancement effect on radio-sensitivity of NK/T cell lynphoma.

Multitargeted tyrosine kinase inhibitors (MTKIs) have been shown to combine with natural killer (NK) cell adoptive transfer for the treatment in various cancers. MTKIs sensitize cancer cells to NK cell therapy through upregulation of nature killer group 2 member D ligands (NKG2DLs) on tumor cells. However, the molecular mechanism of MTKIs-mediated upregulation of NKG2DLs is still unknown. In this study, we confirmed sunitinib induced downregulation of its targets, such as vascular endothelial growth factor, platelet-derived growth factor, and c-kit in multiple-drug–resistant nasopharyngeal carcinoma cell line CNE2/DDP and hepatoma cell line HepG2. Then, we further showed sunitinib induced cell proliferation inhibition, apoptosis, and DNA damage in CNE2/DDP and HepG2 cells. Coculture experiments showed that sunitinib-treated CNE2/DDP and HepG2 cells were able to increase the activation and cytotoxicity of NK cells. Quantitative polymerase chain reaction results showed that sunitinib upregulated NKG2DLs, apoptotic genes, DNA damage repair genes, and nuclear factor (NF)-κβ family genes. Silencing of NF-κβ1, NF-κβ2, or RelB (NF-κβ pathway) inhibited sunitinib-induced upregulation of NKG2DLs. Taken together, we concluded that sunitinib upregulated NKG2DLs through NF-κβ signaling noncanonical pathway which might mediate higher cytotoxic sensitivity of CNE2/DDP and HepG2 cells to NK cells.

Objective:To observe the safety and efficacy through the application of autologous NK cells in MMASCT transplantation.Methods: 8 cases of MM patients, including 7 males and 1 female, aged 42-62 years, the median age of 51 years, before transplantation CR2 cases, PR6 cases.NK cell culture method: Membrane chimeric activity factor culture system.The pretreatment program is based on the standard melphalan or Baihuaan or fludarabine-based regimen.Two of the NK cells were transfused 6-7 days after stem cell infusion, and six of the NK cells were used as part of the transplant pretreatment program, 48 hours after chemotherapy and 24 to 48 hours before the stem cell was returned.Stem cell infusion: CD34 + cells 2.2-4.0x10 ^ 6 / kg, mononuclear cells 4.3-5.8x10 ^ 8 / kg.NK cell infusion: 16-160 * 10 ^ 6 / kg.Observe the adverse reactions, blood cell recovery time and efficacy.Results: 8 cases of fever in 6 patients, body temperature 38-39 degrees, the clinical consideration of lung infection, 1 case of oral mucositis (grade II), anti-infection symptomatic treatment after remission, no rash, diarrhea and other adverse reactions The recovery time of hematopoietic cells were as follows: granulocyte (more than 0.5 * 10 ^ 9 / L continuous 3d) 7-14d, median time 9 days, platelet (greater than 30 * 10 ^ 9 / L continuous 3d) 9-34 days Bit time 11 days.8 patients were successfully completed ASCT.3 months after transplantation, 3 cases of CR, 5 cases of PR.Eight patients were given thalidomide maintenance therapy.The follow-up period was 3 to 29 months, with a median follow-up of 11 months and no deaths during follow-up.Conclusion: Autologous NK cells in the MMASCT transplantation during the application, no inflammatory factor storm effect occurs, return safety, the treatment is effective.To further expand the number of cases of clinical studies to observe its long-term adverse reactions and efficacy.

OBJECTIVE:To explore the effect of arsenic trioxide combined with itraconazole on proliferation and apoptosis of KG1a cells and its potential mechanism.METHODS:The cell morphology was observed with Wrighe-Giemsa staining; cell survival rate was examined by CCK-8; and colony formation capacity was measured by methylcellulose colony formation test; the flow cytometry was used to analyse the cell apoptosis rate and cell cycle; the protein expressions of BCL-2,caspase-3,BAX,SMO,Gli1 and Gli2 were detected by Western-blot.RESULTS:The arsenic trioxide and itraconazole alone both could inhibit the KG1a cell proliferation in dose-and time-dependent manner. In comparison between single and combined drug-treatment group, both the cell survival rate and the colony number of the single drug-treatment group were significantly lower(P<0.05), and the apoptosis rate was higher in the combined drug-treatment group. In the combined-treatment group, the protein expression of Caspase-3 and BAX was upregulated, while the protein expression of BCL-2,SMO,Gli1 and Gli2 was downregulated.CONCLUSION:Arsenic trioxide combined with itraconazole can inhibit the KG1a cell proliferation and induce apoptosis, which may be related with the inhibition of Hh signaling pathway and upregulation of both Caspase-3 and BAX protein expression, and provided experimental data of arsenic trioxide combined with itraconazole for the treatment of refractory AML.

Objective For providing experimental platform of chronic graft-versus-host disease (cGVHD),to establish a mouse model by haplo-identical spleen cell infusion.Methods The donor male mice (Balb/cH-2d) and the recipient (Balb/c C57BL/6) F1 H2-d/b (CB6F1) female mice were randomly divided into four groups:3 experimental groups injected with 3 107,6 107 and 9 107 spleen cells,respectively,while the control group received RPMI 1640 solution.H-2d and H-2b were checked to analyze the chimerism in bone marrow cells.Body mass,figure,cutaneous manifestation and survival of recipient mice were observed and scored every 3 days.Pathologic changes of target organs were observed and scored.Results Injection of 6 107 and 6 107 splenocytes in the recipient mice resulted in a chronic disease with a low level of parental cell engraftment steadily.As compared with 3 107 group,the incidence of cGVHD in 6 107 and 9 107 groups were significantly increased (P ＜0.01).But there was no significant difference between 6 107 and 9 107 groups (P＞0.05).Conclusion A murine model of cGVHD after haplo-identical spleen cell infusion of donor is successfully established by injection of 6 107 and 9 107 spleen cells.

[基金项目] 国家自然科学基金青年项目资助(No.81400156)。Project supported by the National Natural Science Foundation for Youth Scholar of China(No.81400156) [作者简介] 吴少杰(1982-),博士,主治医师,讲师,主要从事淋巴瘤耐药机制的研究,E-mail:wshj510515@163.com [通信作者] 郭坤元(GUO Kunyuan,corresponding author),博士、主任医师、教授,主要从事恶性血液肿瘤的防治基础和临床研究,E-mail:wodeyoujian@foxmail.com [优先发表] http://kns.cnki.net/kcms/detail/31.1725.R.20170731.1054.018.html MicroRNA-150对NK/T细胞淋巴瘤组织和NK-92细胞辐射敏感性的影响

There are few reports with respect to the details of acute effects(during and following RT) of ionizing radiation in liver tissue in epidemiology, pathology, physiology and imageology. Therefore, this study was undertaken to assess the diagnostic value of 2-[ 18 F]-fluoro-2-deoxy-Dglucose positron emission tomography with computed tomography([ 18 F]-FDG-PET/CT) in the detection of acute radiation toxicity in normal liver using Tibet minipigs as a model. Thirty-six male Tibetan minipigs were randomly divided into six groups(n ? 6). The irradiation groups were treated with a single dose of 2; 5; 8; 11 and 14 Gy total body irradiation using an 8-MV X-ray linac, at dose rate of 255 c Gy/min. The control group was not irradiated. The pigs were evaluated with [18F]-FDG-PET/CT, and their alanine aminotransferase(AST) and aspartate aminotransferase(ALT) activities and the ALT/AST ratio, INRatio and bilirubin were measured on Day 7 post-TBI. All pigs were killed on Day 7 post-TBI to collect liver tissues for pathological examination. The results showed that liver standardized uptake value(SUV) increased with the dose from 2 to 11 Gy and decreased suddenly at 14 Gy. HAI score showed a radiation dose-dependent increase of 2–11 Gy and was positively correlated with SUV(p\0:05). However,HAI score showed no correlation with liver function.Therefore, we concluded that [ 18 F]-FDG-PET/CT has the potential to assess acute radiation-induced hepatic injury on early stage.

BACKGROUND:Bone marrow mesenchymal stem cels have low immunogenicity and can induce immune tolerance. At present, the mechanism of immune regulation of bone marrow mesenchymal stem cels is not completely understood. It has been rarely reported whether the bone marrow mesenchymal stem cels can migrate to the thymus after transplantation. <br> OBJECTIVE:To observe the distribution and survival of bone marrow mesenchymal stem cels in the thymus of aging rats after transplantation. <br> METHODS: Bone marrow mesenchymal stem cels cultured in vitrowere transfected by adenovirus vectors expressing green fluorescent protein. Transfected bone marrow mesenchymal stem cels were injected into the portal vein of aging rats. At days 3, 7, 14, 21 after transplantation, the survival of bone marrow mesenchymal stem cels homing to the thymus was observed under fluorescence microscope. At day 3 after transplantation, thymus tissues were taken and stained with hematoxylin-eosin for pathological observation. <br> RESULTS AND CONCLUSION:Green fluorescent protein-labeled bone marrow mesenchymal stem cels had a strong green fluorescence at days 3 and 7 after transplantation, and the cel contour was clear. There was no significant difference in the mean absorbance values at days 3 and 7 (P> 0.05). Expression of green fluorescent protein was weakened significantly at days 14 and 21 compared with that at day 3 (P < 0.05). At 3 days after transplantation, the transplanted bone marrow mesenchymal stem cels were clearly visible in the thymus, and acute rejection was not observed. The results show that bone marrow mesenchymal stem cels can migrate to the damaged thymus tissue through the blood circulation, and can survive at least 1 week.

There are few reports with respect to the details of acute effects (during and following RT) of ionizing radiation in liver tissue in epidemiology, pathology, physiology and imageology. Therefore, this study was undertaken to assess the diagnostic value of 2-[\({}^{18}\hbox {F}\)]-fluoro-2-deoxy-D-glucose positron emission tomography with computed tomography ([\({}^{18}\hbox {F}\)]-FDG-PET/CT) in the detection of acute radiation toxicity in normal liver using Tibet minipigs as a model. Thirty-six male Tibetan minipigs were randomly divided into six groups (\(n = 6\)). The irradiation groups were treated with a single dose of \(2, 5, 8, 11 \hbox { and } 14 \hbox { Gy}\) total body irradiation using an 8-MV X-ray linac, at dose rate of \(255 \hbox { cGy/min}\). The control group was not irradiated. The pigs were evaluated with [\({}^{18}\hbox {F}\)]-FDG-PET/CT, and their alanine aminotransferase (AST) and aspartate aminotransferase (ALT) activities and the ALT/AST ratio, INRatio and bilirubin were measured on Day 7 post-TBI. All pigs were killed on Day 7 post-TBI to collect liver tissues for pathological examination. The results showed that liver standardized uptake value (SUV) increased with the dose from 2 to 11 Gy and decreased suddenly at \(14 \hbox { Gy}\). HAI score showed a radiation dose-dependent increase of 2–11 Gy and was positively correlated with SUV (\(p<0.05\)). However, HAI score showed no correlation with liver function. Therefore, we concluded that [\({}^{18}\hbox {F}\)]-FDG-PET/CT has the potential to assess acute radiation-induced hepatic injury on early stage.

Low response, treatment-related complications and relapse due to the low sensitivity of myelodysplastic syndrome (MDS) and leukemia stem cells (LSCs) or pre-LSCs to arsenic trioxide (ATO), represent the main problems following treatment with ATO alone in patients with MDS. To solve these problems, a chemosensitization agent can be applied to increase the susceptibility of these cells to ATO. Curcumin (CUR), which possesses a wide range of anticancer activities, is a commonly used chemosensitization agent for various types of tumors, including hematopoietic malignancies. In the present study, we investigated the cytotoxic effects and potential mechanisms in MDS-SKM-1 and leukemia stem-like KG1a cells treated with CUR and ATO alone or in combination. CUR and ATO exhibited growth inhibition detected by MTT assays and apoptosis analyzed by Annexin V/PI analyses in both SKM-1 and KG1a cells. Apoptosis of SKM-1 and KG1a cells determined by Annexin V/PI was significantly enhanced in the combination groups compared with the groups treated with either agent alone. Further evaluation was performed by western blotting for two hallmark markers of apoptosis, caspase-3 and cleaved-PARP. Co-treatment of the cells with CUR and ATO resulted in significant synergistic effects. In SKM-1 and KG1a cells, 31 and 13 proteins analyzed by protein array assays were modulated, respectively. Notably, survivin protein expression levels were downregulated in both cell lines treated with CUR alone and in combination with ATO, particularly in the latter case. Susceptibility to apoptosis was significantly increased in SKM-1 and KG1a cells treated with siRNA-survivin and ATO. These results suggested that CUR increased the sensitivity of SKM-1 and KG1a cells to ATO by downregulating the expression of survivin.

Acute graft-versus-host disease (aGVHD) is a major complication of allogeneic hematopoietic stem cell transplantation (allo-HSCT) and a major cause of nonrelapse mortality after allo-HSCT. A conditioning regimen plays a pivotal role in the development of aGVHD. To provide a platform for studying aGVHD and evaluating the impact of different conditioning regimens, we established a murine aGVHD model that simulates the clinical situation and can be conditioned with Busulfan-Cyclophosphamide (Bu-Cy) and Fludarabine-Busulfan (Flu-Bu). In our study, BALB/c mice were conditioned with Bu-Cy or Flu-Bu and transplanted with 2 × 107 bone marrow cells and 2 × 107 splenocytes from either allogeneic (C57BL/6) or syngeneic (BALB/c) donors. The allogeneic recipients conditioned with Bu-Cy had shorter survivals (P < 0.05), more severe clinical manifestations, and higher hepatic and intestinal pathology scores, associated with increased INF-γ expression and diminished IL-4 expression in serum, compared to allogeneic recipients conditioned with Flu-Bu. Moreover, higher donor-derived T-cell infiltration and severely impaired B-cell development were seen in the bone marrow of mice, exhibiting aGVHD and conditioned with Flu-Bu. Our study showed that the conditioning regimen with Bu-Cy resulted in more severe aGVHD while the Flu-Bu regimen was associated with more extensive and long standing bone marrow damage.

目的 采用化疗药物预处理方案建立异基因造血干细胞移植的慢性移植物抗宿主病小鼠模型,并进行评价.方法 以BALB/cH-2kd小鼠作为供鼠,C57BL/6H-2kd小鼠为受鼠.对受鼠使用不同的化疗药物进行预处理[方案1:白消安20mg/(kg·d)×4d+环磷酰胺150mg/(kg·d)×2 d;方案2:白消安20mg/(kg·d)×4 d+环磷酰胺100mg/(kg·d)×2 d],然后经尾静脉注射不同剂量的供鼠脾细胞(6×107或4×107个)和(或)相同剂量的骨髓细胞(2×107个),建立慢性移植物抗宿主病小鼠模型;采用嵌合体分析、临床评分、组织病理学等进行评价.结果 采用白消安20mg/(kg·d)×4d-+环磷酰胺150mg/(kg·d)×2d的方案进行预处理、2×107个骨髓单个核细胞+6×107个脾单个核细胞进行移植可形成较高水平的供受者混合嵌合体.移植物抗宿主病发生时间多集中在供鼠脾细胞输注后30～90 d,化疗药物剂量大的预处理方案及移植细胞数高的移植组小鼠的临床评分和慢性移植物抗宿主病的发生率高于化疗药物剂量小的预处理方案及移植细胞数少的移植组(P＜0.05).模型小鼠肠、肝脏、皮肤、脾脏等器官出现细胞和结构异常、炎症细胞浸润等病理改变.结论 采用白消安和环磷酰胺预处理、给予2×107个骨髓单个核细胞+6×107或4×107个脾单个核细胞可形成较稳定的慢性移植物抗宿主病小鼠模型,为进一步指导临床治疗慢性移植物抗宿主病奠定了实验基础.

Objective To analyze the relationship between cytomegalovirus (CMV) reactivation and leukemia relapse after haploidentical hematopoietic stem cell transplantation (haplo-HSCT).Methods A total of 61 patients diagnosed as hematological malignancies undergoing haplo-HSCT were analyzed retrospectively in our center.Results In the cohort,36 patients had CMV reactivation after haplo-HSCT.The 100-day cumulative incidence of CMV reactivation was 59％.Compared with that in patients without CMV reactivation after transplantation,the incidence of leukemia relapse was lower in patients with CMV reactivation (16.9％ vs 40.0％,P =0.034).The correlation of CMV reactivation and decreased relapse rate was only found in patients with acute myeloid leukemia (P =0.019).In multivariate analysis,relapsed disease status before transplant was a significant negative predictor of overall survival (OS) and relapse after transplant (RR was 2.866 and 3.331 respectively).CMV reactivation after transplant had a protective effect on disease relapse (RR =0.300,P =0.047).Conclusions The rate of CMV reactivation after haplo-HSCT is high.CMV reactivation may reduce risk of relapse in patients diagnosed as acute myeloid leukemia undergoing haplo-HSCT.However,CMV reactivation is one of the important predictors of non-relapse death after transplant,active anti-viral treatment is still needed.

Acute myeloid leukemia (AML) is a common disorder in the elderly. Although remarkable progress has been made over recent decades, the outcome remains poor. Thus, the development of a more effective method to overcome this problem is necessary. In this study, we aimed to investigate the synergistic cytotoxic effect of low-dose arsenic trioxide (As2O3) combined with aclacinomycin A (ACM) on the human AML cell lines KG-1a and HL-60, and to clarify the underlying mechanism. Results showed that As2O3 combined with ACM exerted a synergistic cytotoxic effect by activation of the apoptosis pathway. Additionally, we found that the combination treatment decreased Bcl-2, c-IAP and XIAP expression but increased SMAC and caspase-3 expression more significantly than the single drug treatments. Furthermore, combination index (CI) values were < 1 in all matched combination groups. Additional evaluation of As2O3 combined with ACM as a potential therapeutic benefit for AML seems warranted.