Circulating tumor DNA (ctDNA) was demonstrated as a biomarker for predicting clinical outcomes in B-cell lymphoma (BCL) patients. However, ctDNA and its minimum number of mutant genes that stratify relapsed or refractory BCL (R/R BCL) patients receiving chimeric antigen receptor (CAR)-T cell therapy remains unknown. The plasma ctDNA of 10 patients with R/R BCL receiving CAR-T cell therapy from our clinical center was adopted for targeted exome sequencing. Regarding R/R BCL patients who received CAR-T cell therapy, their 6-, 12- and 18-month overall survival (OS) rates were 90%, 80% and 64%, respectively, while their complete remission (CR) rate was 70%. There was a clear trend suggesting that ctDNA negative was related to favorable OS, even though it was not of significance at that point (P = 0.09). To be specific, BCL2, NRAS, and TBL1XR1 in ctDNA were the optimal combination for predicting OS of R/R BCL patients (P = 0.009). Importantly, ctDNA and BCL2/ NRAS/ TBL1XR1 were all positively correlated with 18 fluorodeoxyglucose positron emission tomography/computed tomography (18 FDG-PET/CT) in evaluating the effectiveness of CAT-T cell therapy (P < 0.05). We identified ctDNA and BCL2/NRAS/TBL1XR1 mutations that can predict the prognosis for R/R BCL patients, which might assist to better select R/R BCL patients for CAR T-cell therapy.

Background: Ferroptosis is a new form of iron-dependent cell death associated with the pathophysiology of vascular dementia (VD). However, the specific mechanism of action is still unclear. This study aims to explore the specific mechanism of action between ferroptosis and VD to find effective therapeutic targets.Methods: Mice were treated with bilateral common carotid artery stenosis (BCAS) to mimic VD models. Neurological function was evaluated using a Morris water maze and mouse hippocampal pathology. Transcriptomic sequencing was performed on hippocampal tissue to search for target genes related to ferroptosis, which was validated in oxygen-glucose deprivation (OGD) treated PC12 cells. Iron content, reactive oxygen species (ROS), and lipid hydroperoxide (LPO) levels were measured. Glu-tathione peroxidase 4 (GPX4) and long-chain acyl-CoA synthetase 4 (ACSL4) levels were measured using WB (Western Blot) and qPCR (Quantitative Polymerase Chain Reaction), respectively.Results: Plasma prekallikrein (KLKB1, PK) levels were elevated in the hippocampal of BCAS-induced mice and OGD-exposed PC12 cells. Compared with the blank group, KLKB1 knockdown protected PC12 cells from OGD-induced apoptosis, and KLKB1 knockdown reduced ROS production and lipid peroxidation in PC12 cells. Besides, KLKB1 knockdown decreased iron content and elevated GPX4 expression.Conclusions: KLKB1 knockdown alleviates cognitive disorder via inhibiting ferroptosis in VD. Inhibition of oxidative stress, lipid peroxidation and reduction of iron content are involved in the underlying mechanisms.

目的 探讨急性脑梗死(ACI)患者血清miR-184的表达水平与病情严重程度、梗死灶体积大小与炎性因子的相关性.方法 纳入蚌埠医学院第一附属医院神经内科2021年1-12月住院的ACI患者74例,健康体检者41例,采用qPCR、ELISA法检测2组血清miR-184、IL-6、TNF-α、CRP表达水平并进行比较,应用Spearman相关分析研究ACI患者miR-184水平与病情严重程度及与炎性因子的相关性.结果 与对照组比较,ACI组miR-184水平降低(P＜0.001),IL-6、TNF-α和CRP水平升高(均P＜0.05),且梗死灶体积越大,NHISS评分越高,miR-184水平越低.Spearman相关性分析结果显示:ACI患者血清miR-184水平与病情严重程度、梗死灶体积呈负相关关系(rs=-0.641、-0.620,均P＜0.05),ACI患者血清miR-184水平与炎性因子IL-6、TNF-α、CRP水平也具有相关性(rs=-0.560、-0.389、-0.565,均P＜0.05).结论 miR-184与脑梗死病情严重程度、梗死灶体积及炎性因子具有相关性,miR-184可以作为诊断ACI及评估病情严重程度的血清学标志物.

目的 探索血浆纤维蛋白原(FIB)及中性粒细胞/淋巴细胞比值(NLR)对Guillain-Barrés综合征(GBS)疾病严重程度的预测价值.方法 选取蚌埠医学院第一附属医院2016年1月至2020年12月收治的109例GBS患者,根据住院免疫治疗前休斯功能分级量表(HFGS)分为轻度组66例及中重度组43例,比较两组年龄、性别、前驱感染史类型、发病至免疫治疗时间、免疫治疗前HFGS评分、血浆白蛋白、三酰甘油、C反应蛋白(CRP)、FIB、NLR等指标的差异.结果 轻度组与中重度组之间年龄、白蛋白、三酰甘油、CRP、FIB、D-二聚体、中性粒细胞计数、淋巴细胞计数、NLR及血小板/淋巴细胞比值差异均有统计学意义(均P<0.05).通过多因素Logstic回归分析血浆FIB和NLR水平是影响GBS患者疾病严重程度的独立危险因素(OR=10.78,P<0.05;OR=2.23,P<0.05).ROC曲线显示,血浆FIB预测中重度GBS的曲线下面积是0.912,最佳截断点是3.61,灵敏度及特异性分别是90.7％、77.3％;NLR预测中重度GBS的曲线下面积是0.856,最佳截断点是4.11,灵敏度及特异性分别是72.1％、90.9％.Spearman相关分析结果显示,血浆FIB、NLR与免疫治疗前HFGS评分呈正相关(r=0.610,P<0.05;r=0.549,P<0.05).结论 血浆FIB、NLR对评估GBS疾病严重程度有良好的预测价值.

【Abstract】 Objective To investigate the efficacy and safety of CD19-targeted chimeric antigen receptor T cell (CAR-T cell) for refractory/relapsed B-cell lymphoma. Methods The efficacy and safety of CD19-CAR-T cells(4-1BB costimulatory domain) in treatment of 12 patients with relapsed/refractory B-cell lymphoma from March 2018 to December 2019 in the Department of Hematology of Guangdong Second Province Hospital were collected analyzed retrospectively. There were 9 patients (75%) with diffuse large B cell lymphoma, 1 patient with blastic variant of mantle cell lymphoma, 1 patient(8.3%) with Burkitt lymphoma, 1 patient with B cell non-Hodgkin lymphoma that cannot be classified. 3 patients (25%) with large mass (≥7.5cm) and 9 patients (75%) with ECOG score ≥2. The number of chemotherapy courses received before transfusion was 4-9, the median number of chemotherapy courses was 7. All 12 patients were autogenous mouse CAR-T cells. Fludarabine + Cyclophosphamide (FC) regimen was used for pretreatment before transfusion, and the number of CAR-T cells was 1 ~ 3.69×10 6/kg. Results All 12 patients received CD19-targeted CAR-T cell therapy. There were 9 patients had treatment response, and the total effective rate was 75%. Among them, there were 3 patients with complete response (CR), with CR rate of 25%, and 6 patients with partial response (PR), with PR rate of 50%. Among the 3 patients with CR remained CR at the follow-up date. Among the 6 patients with PR, 4 showed disease progression in the second month after transfusion, and 2 showed disease progression in the third month after transfusion. All the 9 patients with effective treatment had different degrees of cytokine release syndrome (CRS), including 3 level-1 CRS, 4 level-2 CRS, and 2 level-3 CRS. Two of them had grade 2 CRES, and all CRS and CRES were controlled after treatment with IL-6 receptor antagonists and glucocorticoids. None of the 3 patients failed to respond to treatment had CRS. Conclusion CD19-targeted CAR-T cell immunotherapy has been shown to be effective in CD19-antigen positive B-cell lymphoma, and adverse CRS reactions during treatment can be controlled after treatment. Patients who obtained CR seemed to be able to maintain long-term CR status, while patients who failed to obtain CR showed disease progression within a short period of 3 months, suggesting that patients who obtained CR at an early stage could achieve better efficacy. Therefore, how to identify patients who receive CR at an early stage may be a research direction for the clinical application of CAR-T cell immunotherapy in B-cell lymphoma.

Objective To evaluate the efficacy and safety of the CD19-targeted chimeric antigen receptor T-cell(CD19-CAR-T) therapy for relapsed/refractory B-cell acute lymphoblast leukemia(B-ALL). Methods The efficacy and safety of CD19-CAR-T cells(4-1BB costimulatory domain) in treatment of 34 patients with relapsed/refractory B-ALL from March 2015 to December 2019 in the Department of Hematology of Guangdong Second Province Hospital were collected analyzed retrospectively. There were 18 cases (52.9%) with high-risk cytogenetic or molecular markers, 14 cases (41.2%) with tumor load was ≥50% before transfusion, 24 cases (70.6%) with ECOG score ≥2. The number of chemotherapy courses received before transfusion was 2-15, the median number of chemotherapy courses was 5. There were 32 autogenous CAR-T cells and 2 donor-derived CAR-T cells, 11 of them received allogeneic hematopoietic stem cell transplantation (allo-HSCT) before transfusion. All were mouse CAR-T cells. Fludarabine + Cyclophosphamide (FC) regimen was used for pretreatment before transfusion, and the number of CAR-T cells was 1 ~ 13.4×10 6/kg. Results All 34 patients received CD19-targeted CAR-T cell therapy. 22 patients obtained MRD- after 1 month, CR rate was 64.7%. 20 patients maintained MRD- after 2 months, and the CR rate was 58.8%. 13 patients still maintained MRD- after 3 months, with a CR rate of 38.2%. 4 patients with recurrence presented CD19 negative recurrence. 10 patients underwent Allo-HSCT after CR acquisition, 6 of them maintained a continuous CR state, and 4 patients died after recurrence. Cytokines release syndrome (CRS) was observed in 31 patients (91.2%). Among them, there were 20 patients (64.5%) with grade 1 ~ 2, 8 patients (25.8%) with grade 3 ~ 4, and 3 patients (9.7%) with grade 5. The cytokines levels of IL-6 and IFN-γ were mainly increased in 20 (64.5%) and 18 (58.1%) patients, respectively. Common clinical adverse reactions are: fever with 32 cases (94.1%), pancytopenia with 28 cases (82.4%), chills with 17 cases (50.0%), fatigue with 26 cases (76.5%), hypotension with 27 cases (79.4%), tachycardia with 24 cases (70.6%), hypofibrinogenemia with 20 cases (58.8%), hypoproglobinemia with 27 cases (79.4%), neurotoxicity with 15 cases (44.1%), nausea with 16 cases (47.1%), vomiting with 14 cases (41.2%), hypoalbuminemia with 25 cases (73.5%), transaminase eleations with 16 cases (47.1%), electrolyte metabolic disorders with 27 cases (79.4%) , hypoxemiawith 15 cases (44.1%). Conclusion CAR-T cells therapy is a novel method for the treatment of refractory/recurrent B-ALL with CD19 antigen positive, which can make patients achieved complete remission in a short time, even achieved MRD negative, and most of the CRS appeared in the process of treatment can be controlled by treatment, but the recurrence rate is higher after 3 months later, and can appear CD19 negative relapse. Allo-HSCT as soon as possible after obtaining CR can enable some patients to obtain sustained CR.Therefore, more clinical studies are needed to explore the clinical application of CAR-T cell therapy. Currently, it is believed that bridging with allo-HSCT may be a solution to achieve sustained CR.

Background: We identified the hub genes and pathways dysregulated in acute myeloid leukemia and the potential molecular mechanisms involved. Methods: We downloaded the GSE15061 gene expression dataset from the Gene Expression Omnibus database and used weighted gene co-expression network analysis to identify hub genes. Differential expression of the genes was evaluated using the limma package in R software. Subsequently, we built a protein-protein interaction network followed by functional enrichment analysis. Then, the prognostic significance of gene expression was explored in terms of overall survival. Finally, transcription factor-mRNA (ribonucleic acid) and microRNA-mRNA interaction analysis was also explored. Results: We identified 100 differentially expressed hub genes. Functional enrichment analysis indicated that the genes were principally involved in immune system regulation, host defense, and negative regulation of apoptosis and myeloid cell differentiation. We identified 4 hub genes, the expression of which was significantly correlated with overall survival. Finally, 26 key regulators for hub genes and 38 microRNA-mRNA interactions were identified. Conclusion: We performed a comprehensive bioinformatics analysis of hub genes potentially involved in acute myeloid leukemia development. Further molecular biological experiments are required to confirm the roles played by these genes.

目的:探讨CLAG方案对初治诱导失败和难治复发急性髓系白血病(AML)患者疗效和预后评估.方法:回顾性分析笔者所在医院于2015年1月-2016年1月收治的4例初治诱导失败和6例难治复发AML患者的临床资料,采用克拉屈滨+阿糖胞苷+GCSF进行治疗,对效果和预后进行评估.结果:6例形态学完全缓解,2例部分缓解,2例未缓解,治疗有效率为80％.10例均出现4级白细胞减少、血小板减少和不同部位的感染.治疗后总生存时间为(19.87±3.21)个月,无复发生存时间为(13.27±3.02)个月,随访2年期间4例死亡.结论:CLAG方案对初治诱导失败和难治复发AML患者短期缓解疗效好,但仍应桥接异基因造血干细胞移植以获得长期疗效.

目的:研究苍艾香薰油熏蒸法在无菌层流病房感染控制中的应用效果.方法:选择2015年12月—2017年12月间广东省第二人民医院血液科入住无菌层流病房的72例患者,按照床号先后顺序划分为观察组与对照组各36例,其中1、2、3、4、5号为观察组,6、7、8、9、10号为对照组,对照组采用空气消毒机进行早晚消毒处理,观察组每日采用苍艾香薰熏蒸空气消毒,每周对病房内各个区域采用细菌培养法或ATP荧光检测技术进行检测,统计患者搬离病房保护前院内感染发生率、各物品细菌菌落总数、住院治疗及花费.结果:采用空气消毒机进行早晚消毒处理后,每周同一时间检测,测量病房内空气、物体表面、医护人员手的细菌菌落总数,结果提示观察组各指标值均优于对照组,(t=9.748、12.746、17.775,P<0.05).观察组36例住院患者中,院内感染发生率低、患者层流病房停留时间、治疗费用、抗菌药物使用时间、感染开始及治疗时间均优于对照组,P<0.05.结论:相较于传统空气消毒机处理,在无菌层流病房采用苍艾香薰熏蒸空气消毒法,取得效果较好,可有效控制病房内各物品菌落总数,降低感染率,缩短住院时间.

Objective:To investigate the effects of Artesunate on suppressing the proliferation of Bortezomib-resistant multiple myeloma (MM) cell line and to explore its molecular mechanism on reversal of drug resistance.Method:Human MM cell line NCI-H929 was treated with Bortezomib in a dose-dependent manner to establish Bortezomib-resistant cell line NCI-H929BR.The inhibitory role of Artesunate on NCI-H929BR and its reversal effect against Bortezomib-resistance were determined by methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay.Cell apoptosis was determined by flow cytometry,and the protein expression levels of nuclear factor-κB p65 (NF-κB p65),phospho-nuclear factor-κB p65 (NF-κB p-p65),P-glycoprotein (P-gp),B-cell lymphoma/leukemia-2 (Bcl-2) protein and Bcl-2 associated X protein (Bax) were detected by Western blot assay.Result:Bortezomib-resistance index of NCI-H929BR was 20.2 times.Artesunate treatment had significant inhibitory effect on the proliferation of NCI-H929BR and the inhibitory effect was in a concentration-dependent manner.Bortezomib (50 nmol·L-1) alone had less effect on NCI-H929BR proliferation,while the inhibition rate of artesunate (12.5 mg·L-1) alone was (23.53 ± 2.21)％ (P ＜0.05);Bortezomib combined with artesunate treatment had greater inhibitory effect (60.71 ± 3.43) ％ (P ＜ 0.01).Artesunate treatment increased NCI-H929BR apoptosis,down-regulated NF-κB p65,NF-κB p-p65,P-gp and Bcl-2 expression levels,and upregulated Bax expression level in a concentration-dependent manner.Conclusion:Artesunate could inhibit NCI-H929BR proliferation,promote apoptosis and reverse Bortezomib-resistance,and its mechanism may be associated with down-regulating expression levels of NF-κB p65,NF-κB p-p65,P-gp,Bcl-2 and up-regulating expression level of Bax.

Objective To investigate the effect of imatinib on coagulation function ,platelet parameters and fibrinolytic indexes in patients with chronic myelocytic leukemia (CML) in chronic phase.Methods A total of 112 patients with CML admitted to our hospital between February 2006 and August 2016 were selected as the study group ,and another 112 healthy people who were admitted to our hospital for physical examination were selected as the normal control group at the same time .The study group were treated with imatinib .The platelet parameters ,coagulation function and fibrinolytic indexes were detected before treatment and 3 months after treatment.The adverse reactions were observed and recorded .Results Before treatment,the PLT level was significantly higher in the study group than the control group (P<0.05),and after treatment,PLT levels were significantly decreased (P<0.05).There was no significant difference in PCT ,MPV and PDW between the two groups before and after treatment ( P>0.05 ) .The levels of Fbg and D-D were significantly higher in the study group than the control group before treatment ( P<0.05 ) .After treatment ,the levels of Fbg and D-D decreased significantly ( P<0.05 ) .There was no significant difference in PT , TT and aPTT between the two groups before and after treatment ( P>0.05 ) .The non-hematological adverse reactions were mild , and there were hematological adverse reactions ,such as neutropenia in 57 cases ( 50.89%) and thrombocytopenia in 26 cases(23.21%).After timely decreasing the dosage of imatinib and treatment with G-CSF,the symptoms were relieved .Conclusion The treatment of CML with imatinib can significantly reduce the platelet count and has little effect on platelet morphology .It can restore the levels of Fbg and D-D to normal levels and regulate between coagulation and fibrinolytic system .

目的 探讨青蒿琥酯对耐地塞米松人多发性骨髓瘤(MM)细胞株MM.1R增殖的影响,并探讨其可能作用机制.方法 取对数生长期MM.1S、对塞米松敏感的MM细胞株MM.1R,各分为空白对照组、地塞米松组、青蒿琥酯组、地塞米松+青蒿琥酯组,分别加入等量培养液、20 ng/mL的地塞米松、25μg/mL的青蒿琥酯、20 ng/mL的地塞米松+ 25 μg/mL的青蒿琥酯,观测各组细胞增殖情况,计算细胞增殖抑制率.取MM.1R细胞加入终浓度为25 μg/mL的青蒿琥酯,分别于给药前、给药24 h、给药48 h检测细胞核转录因子κB (NF-κB) p65蛋白、P糖蛋白(P-gp)蛋白的表达.结果 给药24h时,MM.1S细胞地塞米松组、青蒿琥酯组、地塞米松+青蒿琥酯组的细胞增殖抑制率分别为20.41％±2.75％、29.07％ ±2.71％、68.92％ ±4.12％,组间两两比较,P均＜0.05;给药24 h,MM.1R细胞地塞米松组、青蒿琥酯组、地塞米松+青蒿琥酯组的细胞增殖抑制率分别对为4.13％±3.10％、27.43％ ±2.67％、60.14 ±3.24％,组间两两比较,P均＜0.05;青蒿琥酯给药前、给药24 h、给药48 h,MM.1R细胞NF-κB p65蛋白的相对表达量分别为0.89±0.21、0.42 ±0.13、0.23±0.08;P-gp蛋白的相对表达量分别为0.86±0.19、0.43 ±0.13、0.25 ±0.09,随干预时间增加,MM.1R细胞NF-κB p65、P-gp蛋白的相对表达量均降低(P均＜0.05).结论 青蒿琥酯可抑制MM.1S以及MM.1R细胞的增殖,与地塞米松联用有协同效应.青蒿琥酯抑制MM.1R细胞增殖的机制可能为下调NF-κB p65、P-gp蛋白的表达.

Objective To investigate the combination effect of artesunate and lenalidomide on the proliferation of bortezomib-resistant multiple myeloma cell line MM .1R, and its molecular mechanism .Methods MM.1R cells were cultured in RPMI 1640 and treated with DMSO (control group), lenalidomide (20 μmol/L), artesunate (25 μg/mL), or lenalidomide (20μmol/L) plus artesunate (25μg/mL), separately.After 24 or 48 h, the cell proliferation rates were assessed by MTT assay, and the NF-κB (P65), Bcl-xl, Mcl-1, p-Bad, and P-gp protein levels were measured by Western blot .Results The inhibitory rate of cell proliferation was 15.16%by lenalidomide for 24 h, 28.28%by lena-lidomide for 48 h, 45.98%by artesunate for 24 h, 17.19%by artesunate for 24 h, 34.53%by lenalidomide plus artesu-nate for 24 h, or 65.16%by lenalidomide plus artesunate for 48 h (P<0.05).NF-κB (P65), Bcl-xl, Mcl-1, p-Bad, and P-gp proteins were down -regulated by lenalidomide plus artesunate treatment in a time -dependent manner . Conclusion Both artesunate and lenalidomide inhibit MM .1R cell proliferation .Combination of artesunate with lenalido-mide can enlarge this effect through down -regulating NF-κB (P65), Bcl-xl, Mcl-1, p-Bad, and P-gp.

ObjectivesAdministration of anti-CD19 chimeric antigen receptor (CAR)-modified T cells for B-cell malignancies has been remarkably effective in recent clinical trials. To investigate the critical parameters affecting efficacy and evaluated the safety of using CAR T cells targeting CD19 in B-lineage malignancies. We performed a systematic review of reported phase I clinical trials using CAR T cells targeting CD19 in B-lineage malignancies.MethodsWe searched Medline and Embase for studies on anti-CD19 CAR-modified T cells in patients with B-cell malignancies in October 2014. Univariate analyses were performed using the Kaplan-Meier method, and a Cox regression model was used to determine the independent prognostic factors of progression-free survival (PFS).ResultsSix trials involving 50 patients were included in this review. After CAR T-cell infusion, the overall response rate was 48% (complete responses in 24%). The 6-month PFS and 1-year PFS were 43% and 27%, respectively. Statistically significant factors favorably influencing PFS were conditioning chemotherapy (P<0.001), B-cell aplasia (P=0.040), and durable persistence of CAR T cells (P=0.013) in univariate analyses. After multivariate analysis, conditioning chemotherapy remained as an independent prognostic factor for PFS. The most common adverse events were fever, hypotension, rigor, fatigue, bacteremia, chill, dyspnea, and headache, but all were temporary and resolved.ConclusionAnti-CD19 CAR-modified T cells have shown therapeutic efficacy in patients with B-lineage malignancies and were well tolerated in most patients. Conditioning chemotherapy is a prerequisite to improve the clinical outcome.

Rationale:The presence of the Philadelphia chromosome (Ph) in acute lymphoblastic leukemia (ALL) has been associated with a high risk of disease relapse and a poor prognosis. Allogeneic hematopoietic stem cell transplantation (HSCT) is an established treatment for adults with Ph-positive ALL, but relapse remains the primary cause of treatment failure, and is associated with an extremely poor prognosis. The emergence of resistance to tyrosine kinase inhibitors (TKIs) poses a challenge for patients with disease relapses after initial treatment with TKI-containing regimens. Patient concerns:Two patients with TKI-resistant recurrent Ph-positive ALL. Diagnoses:Ph-positive ALL. Interventions:Anti-CD19 CAR T-cell infusion. Outcomes:One patient's bone marrow blasts decreased significantly, and the other reached negative minimal residual disease (MRD). However, we first recorded the development of new-onset acute graft-versus-host disease (aGVHD) after anti-CD19 CAR T-cell infusion in a patient who received allogeneic HSCT. Our 2 case reports also demonstrate the efficacy of anti-CD19 CAR T-cell therapy in the treatment of TKI-resistant Ph-positive ALL. Lessons:Our report suggests that anti-CD19 CAR T-cell therapy may be a promising option for the treatment of relapsed Ph-positive ALL after conventional chemotherapy or allogeneic HSCT. However, caution is due given the possibility of the adverse effects of cytokine release syndrome (CRS)-induced aGVHD for patients receiving allogeneic HSCT.

Objective To investigate the expression level of regulatory T cells(Tregs) in aplastic anemia patients and its changing in the evolution of the disease,and to explore the rule of Tregs on the pathogenesis of apalastic anemia and the possibility of Tregs as the indicator of prognosis and the effect of treatment. Methods Tregs of peripheral blood mononuclear cells were analyzed before and after immunocompressive therapy in 21 patients with severe aplastic anemia by FACS,and,compared with normal control. Results The quantities of Tregs of severe aplastic anemia reduced dramatically compared with normal healthy,and recovered after effective therapy;non response patients remained the level of Tregs unchanged. Conclusion Tregs may play an important role in maintaining normal hematopoiesis,and the reduction of Tregs may be related to the pathogenesis of aplastic anemia.Meanwhile,Tregs could be looked as indicator to monitor the effectiveness of treatment.

Objective To investigate the role of dihydroartemisinin in the apoptosis of human acute myeloblastic leukemia HL-60 cells. Methods The growth inhibition of HL-60 cells treated with dihydroartemisinin was assessed by XTT cell proliferation and cytotoxicity assay kit.The apoptosis and mitochondria membrane potential of HL-60 cells were assessed by flow cytometry with AnnexinV-FITC/PI and JC-1 staining,respectively.The activity of Caspase-3 was measured by ELISA. Results Dihydroartemisinin inhibited HL-60 cells proliferation in a concentration-dependent manner,and induced the apoptosis of HL-60 cells via depolarizing mitochondria membrane and activating Caspase-3. Conclusion Dihydroartemisinin inhibits HL-60 cells proliferation and induces HL-60 cells apoptosis,which may be correlated with mitochondria apoptosis pathway.

Objective: To study the effects of ALLO-NK cell cytotoxicity against leukemic cell lines KG1a treated with Curcumine and to preliminarily explore their molecular mechanisms.Methods: Taking ALLO-NK highly sensitive K562 cells as controls,cytotoxicity of NK cells isolated from 5 healthy volunteers against two leukemia cells was analyzed by LDH releasing assay at different effector-to-target cell ratios(E∶ T).The expressions of NKG2D ligands and HLA-classⅠmolecule were assayed by flow cytometery;the mRNA expression of NKG2D ligands in two kind of cells was measured by RT-PCR;CCK-8 test was conducted to find out the 50 % inhibition concentration of Curcumine and this level of Curcumine was chosen to treat KG1a cells;cytotoxicity of ALLO-NK cells was analyzed by LDH releasing assay and the expressions of NKG2D ligands and HLA-classⅠmolecule were assayed by flow cytometery after treating with Curcumine once again.Results: KG1a cells is not sensitive to ALLO-NK cells.The expressions of NKG2D ligands were positively detected in the two cells at mRNA level,but they were significantly higher in K562 cells than KG1a cells at the protein level(P<0.05).However,the expression of NKG2D ligands in KG1a cells was up-regulated(P<0.05)and no significant difference of expression of HLA-classⅠmolecules was observed after treateing with Curcumine(P>0.05).Conclusion: Curcumine could obviously up-regulate NKG2D ligands expression in KG1a cells and thus enhance the cytotoxicity of NK cells against KG1a cells.

目的观察30例健康供者在应用rhG-CSF动员前后外周血中性粒细胞形态有无改变。方法注射rhG-CSF前后采集静脉血制备血涂片,pH6.4～6.8条件下瑞特-姬姆萨混合染色后在油镜下观察中性粒细胞形态,同时进行中性粒细胞碱性磷酸酶(NAP)染色、pH5.4条件下姬氏染色观察中毒颗粒。结果应用rhG-CSF前1 d中性粒细胞形态无明显异常,中毒颗粒、NAP阳性率及积分值均在正常范围内;应用rhG-CSF后中毒颗粒、NAP阳性率及积分明显增高,96%的中性粒细胞胞体偏大,胞浆内见大量"中毒"性颗粒、D hle小体、空泡数量增加。结论应用rhG-CSF可引起外周血中性粒细胞出现明显的形态变化。

目的探讨减低预处理剂量的异基因造血干细胞移植在难治性急性白血病治疗中的安全性和疗效。方法6例难治性急性白血病,其中5例耐药复发,1例继发性白血病,经常规化疗2~11个疗程未能达到完全缓解。预处理方案为减低预处理剂量的BuCy方案。结果除1例死于肠道Ⅳ度急性GVHD外,其余5例均获得造血重建,各脏器预处理相关毒性(RRT)大多数为Ⅰ~Ⅱ级,未发生严重预处理相关并发症。移植后11~24 d均有经DNA检测证实的完全供者嵌合。发生急性GVHD 4例,2例为Ⅳ度肠道GVHD,分别于+18 d和+64 d死亡。1例为Ⅰ度皮肤GVHD治愈,1例先后出现Ⅳ度皮肤aGVHD和肝脏cGVHD经治疗得到良好控制,无白血病生存。3例移植后发生髓外软组织肿瘤(EMD),采用联合化疗+放疗、供者淋巴细胞输注(DLI),2例进行了二次移植,2例无病生存,1例带瘤生存。结论减低预处理剂量的BuCy方案治疗难治复发性急性白血病,有较好的耐受性,可取得完全型供者细胞植入及血液学缓解,但有可能增加髓外复发的风险。