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To explore the combined pro-apoptosis effect of HSP90 inhibitor BIIB021 and chloroquine (CQ) in chronic myeloid leukemia (CML) cells bearing T315I mutation and its mechanism.The p210-T315I cells were divided into 4 groups by different treatment: control, BIIB021, CQ, and BIIB021 + CQ. After treated with BIIB021 or/and CQ for 24 hours, Annexin V/PI binding assay was used to detect apoptosis rates of CML cells. DAPI staining was used to observe nuclear fragmentation, and Western blot was used to detect the expression of caspase 3, PARP (apoptosis related proteins) and p62, LC3-I/II (autophagy related proteins). P210-T315I cells were inoculated subcutaneously into mice and CML mouse models were established. The mice in treatment groups were injected with BIIB021 and/or CQ while mice in control group were treated with PBS and normal saline. The tumor volume of mice was measured every 4 days, and protein level of cleaved-caspase 3 and LC3-II in tumor tissue were detected by immunohistochemistry.The results showed that BIIB021 induced apoptosis of CML cells in a dose-dependent manner ( r=0.91). CQ could enhance the apoptosis-inducing effect of BIIB021. Flow cytometry analysis results showed that the apoptosis rate of p210-T315I cells in combination group was higher than that in BIIB021 or CQ only group (P0.05). DAPI staining showed nuclear fragmentation in combination group could be observed more obviously. Western blot analysis showed that BIIB021 could induce LC3-I to convert to LC3-II and decrease p62 protein levels (P0.05). Moreover, the combination group had higher expression of LC3-II, p62 (P0.05), activated PARP and activated caspase 3 than BIIB021 only group (P0.05). Besides, experiment in vivo showed the mean tumor volume in co-treatment group was lower than that in single drug group (P0.01). Immunohistochemistry of tumor tissue also showed the protein level of cleaved-caspase 3 and LC3-II in combined group was higher than that in BIIB021 only group.HSP90 inhibitor BIIB021 induced significant apoptosis of CML cells bearing T315I both in vivo and in vitro. CQ can enhance this effect probably by autophagy inhibition.氯喹增强BIIB021对T315I突变的慢性粒细胞白血病细胞促凋亡的研究.探讨氯喹（CQ）联合热休克蛋白90抑制剂BIIB021对T315I突变的慢性粒细胞白血病（CML）p210-T315I细胞凋亡的作用及相关机制。.将p210-T315I 细胞株按不同处理分为对照、BIIB021、氯喹、BIIB021+氯喹共4组。经过BIIB02和/或CQ处理24 h后，流式细胞术检测细胞凋亡，DAPI染色观察细胞核形态及凋亡情况；Western blot检测凋亡相关蛋白caspase 3、PARP，自噬相关蛋白p62、LC3-I/II的表达情况。皮下注射p210-T315I细胞，构建CML小鼠荷瘤模型，模型建立后治疗组分别给药（BIIB021和/或氯喹），对照组给PBS和生理盐水，每4 d观察记录荷瘤小鼠局部肿瘤体积，免疫组织化学法检测瘤体组织cleaved-caspase 3和LC3-II表达水平。.BIIB021有促p210-T315I细胞凋亡的作用，随着药物浓度升高，细胞凋亡率越高（r=0.91）。氯喹能增强BIIB021相关的促凋亡作 用，流式细胞术检测结果显示，联合用药组p210-T315I细胞凋亡率明显高于BIIB021、氯喹单用药组（P0.05）。DAPI染色显示，与药物单用组比较，药物联合应用组细胞中点状浓染更多，细胞核碎裂更明显。凋亡和自噬相关蛋白检测显示，与对照组比较，BIIB021单药组细胞中LC3-I向LC3-II转变增加，p62减少（P0.05）。当氯喹联合BIIB021时，与BIIB021单用相比，LC3-II和p62的表达量增多（P0.05），此时PARP和caspase 3的激活带增多（P0.05）。动物实验显示两种药物联合使用与药物单用相比，小鼠肿瘤体积明显变小（P0.01）。免疫组织化学显示氯喹+BIIB021联用组肿瘤组织中cleaved-caspase 3、LC3-II表达量高于BIIB021单用组。.热休克蛋白90抑制剂BIIB021对T315I突变的CML细胞、小鼠模型有促进凋亡的作用，氯喹能增强这一效应，其机制可能与抑制自噬有关。.

To investigate the effects of dasatinib on the maturation of monocyte-derived dendritic cells (moDCs) derived from healthy donors (HDs) and chronic myelogenous leukemia (CML) patients.Peripheral blood mononuclear cells (PBMCs) were isolated from HDs (n=10) and CML patients (n=10) who had got the remission of MR4.5 with imatinib treatment. The generation of moDCs from PBMCs was completed after 7 days of incubation in DC I culture medium, and another 3 days of incubation in DC II culture medium with or without 25 nmol/L dasatinib. On the 10th day, cells were harvested and expression of molecules of maturation related marker were assessed by flow cytometry. The CD80The proportion of CD80For CML patients, the same high proportion of moDCs as HDs can be induced in vitro, which provides a basis for the application of DC-based immunotherapy strategy. Dasatinib at the concentration of 25 nmol/L can efficiently promote the maturation of moDCs derived from HDs and CML patients in vitro. Dasatinib shows potential as a DC adjuvant to be applied in DC-based immunotherapy strategies, such as DC vaccine and DC cell-therapy.达沙替尼对健康供者和慢性髓细胞白血病患者单核细胞衍生的树突状细胞成熟的影响.探讨达沙替尼对健康供者和慢性髓细胞白血病（CML）患者单核细胞衍生的树突状细胞（moDC）成熟的影响.从健康供者（n=10）和通过伊马替尼治疗获得MR4.5缓解状态的CML患者（n=10）外周血中分离出单个核细胞，并体外培养、诱导形成moDCs。此过程由DC I 培养基培养7 d，以及随后DC II培养基培养3 d完成，其中后3 d给予达沙替尼处理或设对照组。 D 10收获细胞并用流式细胞术分析DCs成熟相关标志物分子的表达。设门总细胞中CD80CD80对于CML患者，体外可诱导出与健康供者同样高比例的moDCs，这为CML患者应用以DC为基础的免疫治疗策略提供了依据。本研究中，在体外25 nmol/L达沙替尼可有效促进健康供者和CML患者来源的moDCs成熟。达沙替尼具有作为一种DC佐剂应用于以DC为基础的免疫治疗策略中的潜力，如DC疫苗和DC细胞治疗.

To evaluate the efficacy of the second-line nilotinib and third-line dasatinib on chronic myelogenous leukemia (CML) with failed first- and second-line treatments, and analyze the influencing factors of the efficacy.Selected 83 patients in The Third People's Hospital of Kunshan City, Jiangsu Province with CML who were treated with nilotinib as the second-line treatment after the failure of the first-line treatment with imatinib as the second-line treatment group (referred to as the second-line group) from January 2014 to December 2018, and 61 CML patients who were treated by dasatinib as the third-line treatment group (referred to as the third-line group) after the failure of the second-line treatment with nilotinib; the first-line treatment with imatinib failed, but due to various reasons, the patients were fully after being informed of the possible serious consequences of not changing the drug treatment, 37 CML patients who were still required to continue imatinib treatment served as the control group. The hematological, genetic and molecular responses of each group were compared for 3, 6, and 24 months of treatment. LogistiC regression was used to analyze the factors affecting the second and third line curative effects.The three groups had statistically significant differences in the rates of achieving CHR, MCyR, and MMR at 3, 6, and 12 months of treatment (P0.05). Compared the two groups, the CHR rates of the second-line group at 3, 6, and 12 months of treatment were 100.00%, 97.59%, and 95.18%, respectively; higher than the third-line group's 90.16%, 86.89%, 83.61% and the control group's 83.78%, 75.68% and 72.97%; the CHR rate of the third-line group was higher than that of the control group at 6 and 12 months of treatment. The rates of reaching MCyR at 3, 6, and 12 months after treatment in the second-line group were 87.95%, 93.98% and 93.98%, respectively, while those in the third-line group were 80.33%, 88.52% and 86.89%, which were higher than those of the control group of 67.57%, 64.86% and 48.65%. The rates of achieving MMR at 3, 6, and 12 months of treatment in the second-line group were 19.28%, 33.72% and 60.24%, respectively, and those in the third-line group were 11.48%, 26.23% and 49.18%, which were higher than those of the control group of 0.00%, 2.70% and 0.00%; The rate of reaching MMR within 12 months of treatment in the second-line group was higher than that of the third-line group, and the differences was statistically significant (P0.05). There was no significant difference in the rate of reaching MCyR between the second-line group and the third-line group at 3, 6, and 12 months, and the rate of reaching MMR at 3 and 6 months (P0.05). The incidence of nausea and vomiting among the three main non-hematological adverse reactions, and the incidence of grade 1~2 anemia among the hematological adverse reactions were statistically significant (P0.05). There was no significant difference in the incidence of rash, eyelid edema, diarrhea, thrombocytopenia, leukopenia and neutropenia in the three groups (P0.05). The incidence of nausea and vomiting and grade 1~2 anemia in the second-line group and the third-line group were higher than that of the control group, and the difference was statistically significant (P0.05). There were statistically significant differences in Sokal score, medication compliance, and hematological adverse reactions between the MMR group and the non-MMR group (P0.05). Logistic regression analysis showed that dose reduction or withdrawal during the treatment period, and grade 3~4 hematological adverse reactions were the main factors affecting the second and third line curative effects (OR=22.160, 2.715, 95% CI=2.795-93.027, 1.882-48.834).The second-line nilotinib and the third-line dasatinib have a better effect on CML patients who have failed the first and second-line treatments. Grade 3~4 hematological adverse reactions, dose reduction or withdrawal are risk factors that affect the efficacy of second and third-line treatments.尼洛替尼二线与达沙替尼三线对一二线治疗失败的 慢性髓性白血病的疗效及影响因素分析.评价尼洛替尼二线与达沙替尼三线对一二线治疗失败的慢性髓性白血病（CML）的疗效，并分析疗效的影响因素.将江苏省昆山市第三人民医院2014年1月-2018年12月伊马替尼一线治疗失败后采用尼洛替尼二线治疗的83例CML患者设为二线治疗组（简称二线组），尼洛替尼二线治疗失败后采用达沙替尼三线治疗的61例CML患者作为三线治疗组（简称三线组）；另选伊马替尼一线治疗失败，但由于多种原因，患者在充分告知不更换药物治疗可能发生的严重后果后，仍要求继续伊马替尼治疗的37例CML患者作为对照组。比较各组患者治疗3、6、24个月的血液学、细胞遗传学和分子学反应。应用LogistiC回归分析影响二三线疗效的因素.三组患者治疗3、6、12个月达完全血液学反应（CHR）、主要细胞遗传学反应（MCyR）、主要分子学反应（MMR）率比较差异均有统计学意义（P0.05）。组间两两比较，二线组治疗3、6、12个月达CHR率分别为100.00%、97.59%、95.18%，高于三线组的90.16%、86.89%、83.61%和对照组的83.78%、75.68%、72.97%；三线组患者治疗6、12个月达CHR率高于对照组。二线组患者治疗3、6、12个月后达MCyR率分别为87.95%、93.98%、93.98%，三线组分别为80.33%、88.52%、86.89%，均高于对照组的67.57%、64.86%、48.65%。二线组患者治疗3、6、12个月达MMR率分别为19.28%、33.72%、60.24%，三线组分别为11.48%、26.23%、49.18%，均高于对照组的0.00%、2.70%、0.00%；二线组治疗12个月达MMR率高于三线组，差异均有统计学意义（P0.05）。二线组与三线组患者治疗3、6、12个月达MCyR率，3、6个月达MMR率差异无统计学意义（P0.05）。三组患者主要非血液不良反应中恶心呕吐发生率、血液学不良反应中1~2级贫血发生率差异有统计学意义（P0.05）。三组的皮疹、眼睑水肿、腹泻、血小板减少、白细胞减少和中性粒细胞减少发生率差异无统计学意义（P0.05）。组间两两比较，二线组、三线组恶心呕吐、1~2级贫血的发生率高于对照组，差异有统计学意义（P0.05）。达MMR组与非MMR组Sokal积分、服药依从性、血液学不良反应比较差异有统计学意义（P0.05）。Logistic回归分析显示，治疗期间减量或停药、3~4级血液学不良反应是影响二、三线疗效的主要因素（OR=22.160，2.715，95%CI=2.795-93.027，1.882-48.834）.尼洛替尼二线与达沙替尼三线对一二线治疗失败的CML患者有较好的疗效。3~4级血液学不良反应、减量或停药是影响二、三线治疗疗效的危险因素.

To investigate the effect of silencing DNA methyltransferase 1(DNMT1) to the methylation of the promoter of the tumor suppressor gene wnt-1 (WIF-1) in human chronic myeloid leukemia (CML) cells.DNMT1 siRNAi plasmid was constructed and DNMT1 siRNAi was transfected into CML K562 cells. RT-PCR and Western blot were used to detect the expression of DNMT1 gene and related protein, and methylation PCR was used to detect WIF-1 gene promoter methylation level. The trypan blue exclusion and MTT assay were used to detect the cell proliferation, flow cytometry were used to detect the cell apoptosis rate, colony formation assay was used to detect cell colony formation ability. Expression of Wnt/β- catenin and its downstream signaling pathway proteins were detected by Western blot after DNMT1 gene was silenced.The expression level of DNMT1 mRNA and its related protein in the experimental group were significantly lower than those in the control group and negative control group (P0.05). After 72 hours of successful transfection, the WIF-1 gene in the control group and negative control group were completely methylated, while in the experimental group, the methylation level significantly decreased. The results of MSP showed that the PCR product amplified by the unmethylated WIF-1 primer in the experimental group increased significantly,while by the methylated WIF-1 primer decreased significantly. After 48 h of transfection, the OD value, viable cell number and colony formation of the cells in experimental group were significantly lower than those in the negative control group and the control group (P0.05). The apoptosis rate of the cells in experimental group was significantly higher than those in the negative control group and control group (P0.05). The expression levels of β- actin, myc, cyclin D1 and TCF-1 in K562 cells in the experimental group were significantly lower than those in the negative control group and control group (P0.05).Silencing DNMT1 gene can inhibit the proliferation and promote the apoptosis of K562 cells. The mechanism may be related to reverse the hypermethylation level of the WIF-1 gene promoter, thereby inhibit the activity of the Wnt/β- catenin signaling pathway.沉默DNMT1减弱WIF-1基因启动子甲基化对慢性髓系白血病K562细胞生物学行为的影响.探讨沉默DNA甲基转移酶（DNMT1）对人慢性髓系白血病细胞内抑癌基因wnt抑制因子-1（WIF-1）基因启动子甲基化的影响。.构建DNMT1 siRNAi质粒，将DNMT1 siRNAi转染至慢性髓系白血病K562细胞中，采用RT-PCR和Western blot法检测DNMT1基因和相关蛋白的表达，并利用甲基化PCR检测WIF-1基因启动子甲基化水平，台盼蓝拒染法和MTT 法检测细胞的增殖能力，流式细胞术检测细胞的凋亡率，集落形成实验检测细胞的集落形成能力，Western blot法检测沉默DNMT1基因后 Wnt/β-catenin及其下游信号通路蛋白的表达情况。.实验组DNMT1 mRNA和相关蛋白表达水平显著低于对照组和阴性对照组（P0.05）；成功转染72 h后，对照组和阴性对照组WIF-1基因处于完全甲基化状态，而实验组WIF-1基因启动子区域甲基化水平显著降低。MSP检测结果显示，实验组未甲基化的WIF-1引物扩增出的PCR产物含量显著增加，而甲基化的WIF-1引物扩增出的PCR产物含量显著降低。转染成功48 h后，实验组的OD值、活细胞数及集落形成数均显著低于阴性对照组和对照组（P0.05）。实验组的细胞凋亡率显著高于阴性对照组和对照组（P0.05）。实验组K562细胞中β-actin、myc、cyclin D1和TCF-1蛋白表达水平均显著低于阴性对照组和对照组（P0.05。.沉默DNMT1基因能够抑制 K562 细胞增殖，促进其凋亡，其作用机制可能与逆转WIF-1基因启动子高甲基化水平，从而抑制Wnt/β-catenin信号通路的活性有关。.

To observe the curative efficacy of tyrosine kinase inhibitors (TKIs) in the treatment of e19a2 transcript (P230) CML chronic phase (CML-CP) patients.The clinical data of 11 P230 CML-CP patients were collected from July 2008 to December 2019. Blood routine examination, bone marrow cytology, chromosome, and BCR-ABL qualitative and quantitative tests were performed at initial diagnosis. After TKIs treatment, BCR-ABL (P230)/ABL in peripheral blood was regularly detected to evaluate molecular response by real-time quantitative PCR.There were 11 patients (7 males and 4 females) in chronic phase from 6 domestic hospitals enrolled, their median age was 46 years old (range from 19 to 56 years old). Among 4 patients treated with imatinib (400 mg, qd) firstly, 3 cases switched to nilotinib (400 mg, bid) and 1 case switched to dasatinib (100 mg, qd) due to failure to achieve best molecular response at the landmark time or mutation of ABL kinase. Then major molecular response (MMR) was obtained within 1 year. In addition, 5 patients were treated with nilotinib (300 mg, bid) and 2 patients with dasatinib (100 mg, qd) as first-line treatment, all of them got MMR within 6 months.For intolerance or resistance to imatinib, second-generation TKIs can enable P230 CML patients to achieve deeper molecular response, and MMR in a short time.酪氨酸激酶抑制剂治疗P230慢性髓系白血病临床疗效观察.观察酪氨酸激酶抑制剂（TKI）治疗e19a2转录本（P230）慢性髓系白血病（CML）慢性期患者的疗效。.收集2008年7月至2019年12月11例P230 CML慢性期患者临床资料，初诊时进行血常规、骨髓细胞学、染色体和BCR-ABL定性和定量检测，接受TKI治疗后，定期通过实时定量PCR方法检测外周血BCR-ABL（P230）/ABL评估分子学反应。.来自国内6家医院11例患者中男性7例，女性4例，中位年龄为46（19-56）岁。11例患者均处于慢性期，4例先接受伊马替尼（400 mg，qd）治疗的患者，由于在规定的时间界点未达到最佳分子学反应或出现ABL激酶区突变，其中3例改为尼洛替尼（400 mg，bid）治疗，1例改为达沙替尼（100 mg，qd）治疗，之后均在1年内获得主要分子学反应。另外5例使用尼洛替尼（300 mg，bid）、2例使用达沙替尼（100 mg，qd）作为一线治疗的患者在6个月内获得了主要分子学反应。.对伊马替尼不耐受或疗效不佳的P230 CML患者，二代TKI可使其达到更深层次分子学反应，且能在更短时间达到主要分子学反应。.

To investigate the regulation of chronic myelogenous leukemia （CML） imatinib resistant genes, in order to improve the therapeutic effect of CML imatinib resistant patients.The human CML cell line K562 and imatinib-resistant K562 cells (K562/G01) were collected, and transcriptome of the cells were achieved by RNA-seq. The sequencing data were analyzed by using standard procedures.Compared with K562 cells, 464 genes were significantly changed in K562/G01 cells, including 163 up-regulated and 301 down-regulated genes. The GO function annotation analysis and KEGG pathway analysis results showed that the differentially expressed genes were mainly involved in biological processes such as oxidative phosphorylation, localization to protein organelle, ribonucleoprotein complex biogenesis and so on. Gene Set Enrichment Analysis (GSEA) plots showed that 5 gene-sets were up-regulated in K562/G01 significantly, including the pathway of TGF-beta, mTOR and CML.CML imatinib resistance is associated with oxidative phosphorylation, during which the pathway of TGF-beta and mTOR are significantly up-regulated.慢性粒细胞白血病耐伊马替尼细胞系的转录组分析.探讨调控慢性粒细胞白血病（CML）耐伊马替尼的基因，以提高CML耐伊马替尼患者的治疗效果.选取伊马替尼敏感细胞系K562和耐药细胞系K562/G01，采用RNA-seq方法获得对应细胞的转录组，并进行标准生物信息学分析.相较于K562细胞，耐伊马替尼细胞K562/G01转录组中共有464个表达显著变化的基因，其中163个基因表达上调，301个基因表达下调。GO功能注释分析和KEGG通路的富集分析结果显示，差异基因主要集中在氧化磷酸化、蛋白细胞器定位、核糖核蛋白复合体的生物发生等生物学过程。GSEA基因富集分析表明，有5个基因集在K562/G01中被显著的上调，如TGF-beta信号通路、mTOR信号通路、慢性粒细胞白血病相关信号通路等.CML细胞伊马替尼耐药与细胞中氧化磷酸化过程有关，在耐药过程中TGF-beta信号通路、mTOR信号通路发生显著上调.

To investigate the clinical characteristics of the patients with chronic myeloid leukemia (CML) discontinued tyrosine kinase inhibitors (TKI) therapy and the outcome of the patients.35 cases of CML patients experienced initiative discontinuation of TKI therapy in our hospital from June 1st 2015 to December 31th 2019 were retrospectively analyzed. The TFR of the patients and the factors affecting it were analyzed.The median duration of TKI administration was 72 (range 35-173) months in the 35 patients. Among these patients, 8 had experienced TKI dose reduction or suspension. All the enrolled patients have achieved at least MMR. The median time for these patients achieving MMR was 15 (range 3-75) months after administration of TKI, and for MMR maintenance before TKI suspension was 55 (range 13-164) months. After TKI withdrawal the median follow up time was 20.3 (range 3-57.9) months, 22 out of 35 patients kept TFR, among them, 2 （5.71%） patients restarted TKI after 12 month suspension, and maintained MMR during suspension. 13 （37.1%）patients lost MMR, among them, 9 patients restarted TKI treatment, and 5 of them achieved MR4.0 after the median duration of 3(2-5) month. No patients were found to have disease progression. The estimated TFR rate was 57.8% and 51.8% at 12 and 24 months after discontinuation, respectively. Other clinical characteristic related to relapse were also analyzed, including the cumulative TKI administration duration, cumulative MMR duration, time to achieve MMR, median age at diagnosis, risk stratification by Sokal score, TKI dose reduction and discontinuation history, and second-generation TKI administration before stopping TKI, however, no statistical difference was found.TKI discontinuation is practical for CML patients in our center.慢性髓细胞白血病患者酪氨酸激酶抑制剂的主动停药临床结局的回顾性分析.研究单中心慢性髓细胞白血病（CML）酪氨酸激酶抑制剂（TKI）治疗后自动停药患者的临床特征及转归情况.回顾性分析2015年6月1日至2019年12月31日在本院长期随访并主动停用TKI的35例CML患者的临床资料，分析患者主动停药后无治疗缓解（TFR）的获得情况及其影响因素.35例患者累积服用TKI中位时间为72（35-173）个月，其中8例患者有减停药史。全部患者在停药前获得主要分子学缓解（MMR）及以上疗效。患者达到MMR中位时间为开始服用TKI后15（3-75）个月，停药前MMR维持时间为55（13-164）个月。停药后中位随访20.3（3-57.9）个月，22例（62.8%）患者获得TFR，其中2例（5.71%）患者因生育停用TKI 12个月以上后重启治疗，停药期间维持MMR。13例（37.1%）患者失去MMR，9例停药后失去MMR的患者再启动药物治疗，其中5例重启后已获得MR 4.0以上疗效，中位时间为 3（2-5）个月。随访全部患者，未发现疾病进展。本组患者经统计学分析显示，12个月TFR率为57.8%，24个月TFR率为51.8%。本研究比较了中位累积TKI治疗时间、中位累积MMR时间、达到MMR时间、中位诊断年龄、Sokal积分、是否具有TKI减停史、停药前使用二代TKI，未发现与分子学复发相关具有统计学差异的临床参数.本中心达停药标准的CML患者能够安全停用 TKI.

To explore the expression level of ETV6-ABL fusion gene in different cell populations in patients with myeloproliferative neoplasm (MPN) and therapeutic effect of tyrosine kinase inhibitor (TKI).A 42-year-old man who presented with fever, marked leukocytosis and chronic myelogenous leukemia (CML) like MPN was reported. ETV6-ABL fusion gene was detected by real-time PCR and confirmed by direct sequencing. ETV6-ABL mRNA expression in each cell population sorted from peripheral blood by flow cytometry was detected by real-time PCR.ETV6-ABL fusion gene was found out in bone marrow cells and confirmed as type A by direct sequencing. ETV6-ABL fusion gene transcript level in polymorphonuclear cells was nearly 3.6-fold relative to that in total cells, which was significantly higher than that in T cell, B cell and monocyte subsets. The complete blood count (CBC) returned to normal level after treatment with imatinib (400 mg) daily for three months. After TKI treatment for 6 months, the ratio of ETV6-ABL/ABL decreased from 174.1% to 1.9%.ETV6-ABL fusion gene positive MPN may have a CML clinical presentation and is sensitive to TKI. ETV6-ABL fusion gene is specifically expressed in polymorphonuclear cells.ETV6-ABL融合基因在骨髓增殖性肿瘤造血细胞群中分布的研究.探讨ETV6-ABL融合基因在不同细胞群体中的表达情况，以及酪氨酸激酶抑制剂对该融合基因的治疗效果.报道了1例42岁的患者，表现为粒细胞增多和慢性粒细胞白血病样骨髓象。使用real-time PCR方法检测融合基因，并测序确认。通过流式分选和real-time PCR方法，对分选后的外周血细胞各个细胞群中的ETV6-ABL mRNA表达水平进行检测.在骨髓细胞中检测到ETV6-ABL融合基因，测序结果显示为ETV6-ABL A型。与总体细胞相比，分叶核细胞中的ETV6-ABL转录水平接近总体的3.6倍，明显高于T细胞、B细胞、单核细胞群。给予伊马替尼每天400 mg治疗3个月后，该患者的血常规恢复到正常水平。伊马替尼治疗6个月后，ETV6-ABL/ABL的比例从最开始的174.1%下降到1.9.ETV6-ABL融合基因阳性的骨髓增殖性肿瘤临床可以表现为慢性粒细胞样，对酪氨酸激酶抑制剂治疗敏感。ETV6-ABL融合基因主要表达在分叶核细胞群.

To investigate the effect of etoposide (ETO) on elimination of chronic myeloid leukemia (CML) stem cells by imatinib mesylate(IM) in vivo.SCL-tTA/BCR-ABL mice were used as CML animal model. Flow cytometry was used to assess the effect of ETO alone or in combination with IM on the number of leukemia stem cell （LSC） in bone marrow and spleen, and peripheral blood neutrophils in CML mice and normal control FVB mice.The results showed that in CML mice, the number and proportion of LSC in bone marrow and the proportion of neutrophils in peripheral blood decreased significantly after ETO and IM combined treatment, and the degree of decrease was more significant than that of both alone. While in wild type FVB mice, the combination of ETO and IM showed no significant effect on the number and proportion of LSK cells in bone marrow and the proportion of neutrophils in spleen.ETO can selectively enhance elimination of CML LSC by IM in vivo.依托泊苷对伊马替尼体内清除慢性髓性白血病干细胞的作用研究.探讨依托泊苷（ETO）对伊马替尼体内清除慢性髓性白血病干细胞的作用.以SCL-tTA/BCR-ABL小鼠为慢性髓性白血病动物模型，采用流式细胞术检测ETO单独或联合伊马替尼治疗慢性髓性白血病小鼠和正常FVB小鼠后，对小鼠骨髓和脾脏中白血病干细胞和外周血中性粒细胞数目的影响.ETO和伊马替尼联合治疗后，SCL-tTA/BCR-ABL小鼠骨髓白血病干细胞数及比例、外周血中性粒细胞比例均明显下降，且下降的程度比两者单用更显著。在野生型FVB小鼠中，ETO和伊马替尼联合治疗对骨髓LinETO在体内具有选择性增强伊马替尼靶向清除慢性髓性白血病干细胞的作用.

To evaluate the clinical efficacy and safety of domestic imatinib (made in China) in patients with newly diagnosed chronic myeloid leukemia chronic phase(CML-CP).Fifty-seven newly diagnosed CML-CP patients who did not receive any other anti-CML treatment were treated by domestic imatinib 400 mg once a day. The hematological, cytogenetic and molecular reactions and safety were observed and evaluated after 3, 6 and 12 months of treatment.Fifty-six patients were treated for ≥3 and 6 months, among which 50 patients were treated for ≥12 months. After 3 months of treatment, 49 patients underwent hematological examination, 47 patients (95.9%) achieved complete hematological response (CHR), 49 patients underwent cytogenetic examination, 39 patients (79.6%) achieved major cytogenetic response (MCyR), and 12 patients (24.5%) achieved complete cytogenetic response (CCyR). 49 patients underwent the level of BCR-ABL test, including 41 patients (83.7%) with BCR-ABLIn the real world, Domestics imatinib mesylate is effective and safe in the treatment of newly diagnosed CML-CP patients, but long-term follow-up data are still necessary to verify its long-term efficacy.真实世界中国产伊马替尼治疗初诊慢性粒细胞白血病的疗效观察.评价国产伊马替尼对初诊慢性粒细胞白血病（CML）慢性期（CP）患者的临床疗效及安全性.57例初诊且未接受其他任何抗CML治疗的CML-CP患者，给予口服国产伊马替尼治疗，400 mg/d，每日1次，观察评估治疗3、6、12个月的血液学、细胞遗传学和分子学反应及安全性.治疗≥3个月和6个月均有56例患者，其中50例患者治疗≥12个月。治疗满3个月时，49例患者进行了血液学检测，47例(95.9％)获得完全血液学反应（CHR）；49例患者进行了细胞遗传学检查，39例(79.6％)获得主要细胞遗传学反应（MCyR）,12例(24.5％)获得完全细胞遗传学反应（CCyR）；49例患者进行了BCR-ABL水平检测，其中BCR-ABL在真实世界中，国产甲磺酸伊马替尼治疗初诊CML-CP患者的近期疗效肯定，安全性可靠，但仍需大宗数据进行长期追踪随访来验证其远期疗效.

To study the effect of 5-aminoimidazole-4-formamide ribonucleotide (AICAR) combined with interferon (IFN-α-2b) on the proliferation and apoptosis of chronic myeloid leukemia K562 cells, and explore its possible mechanism.CCK-8 method was used to detect the inhibition of cell proliferation. Wright Giemsa method was used to stain and cell morphology was observed by light microscopy. FITC Annexin V/PI double staining method was used to analyze the change of apoptosis rate. Immunocytochemistry method was used to detect the expression of wild-type P53 protein.Different concentration of AICAR was inhibitory effect on K562 cells at different time point of action for 24 h, 48 h, and 72 h, and the inhibition was time and dose-dependent (r=0.71, r=0.84). The combination of AICAR and IFN-α-2b could effectively inhibit the proliferation and promote apoptosis of K562 cells. The inhibition rate of K562 cells was (45.26±2.54)%, and the early apoptosis rate was (33.72±0.23)%, which was statistically significantly different from the control group, AICAR or IFN-ɑ-2b alone (P0.05). The combination of two drugs promoted the expression of wild-type p53 protein.AICAR and/or IFN-ɑ-2b can inhibit the cell proliferation and promote the apoptosis of K562 cells. The combination of two drugs shows synergistic antitumor effect, and its mechanism may be related to the promotion of high expression of wild-type p53 protein.5-氨基咪唑-4-甲酰胺核糖核苷酸联合干扰素对K562细胞的影响.研究5-氨基咪唑-4-甲酰胺核糖核苷酸（AICAR）联合干扰素（IFN-ɑ-2b）对慢性粒细胞白血病K562细胞的增殖及凋亡的影响，并探讨其可能的作用机制.采用CCK-8法检测细胞增殖抑制情况；Wright-Giemsa方法染色，光学显微镜下观察细胞形态；FITC AnnexinV/PI双染法分析细胞凋亡率的变化；免疫细胞化学法检测野生型P53蛋白的阳性表达率.不同浓度AICAR于不同作用时间24、48和72 h对K562细胞均有抑制作用，抑制程度呈时间、剂量依赖性（r=0.71，r=0.84）。AICAR与IFN-ɑ-2b联合应用后可以有效抑制K562细胞增殖，促进细胞凋亡，联合作用72 h时细胞的抑制率可达（45.26±2.54）％，联合作用48 h早期凋亡率达（33.72±0.23）％，与对照组及单用AICAR和单用IFN-ɑ-2b相比，差异均有统计学意义（P0.05）。AICAR与IFN-ɑ-2b联合作用促进野生型P53蛋白的表达.AICAR和（或）IFN-ɑ-2b能抑制细胞增殖，促进K562细胞凋亡，2药联合具有协同抗瘤效应，其作用机制可能与促进野生型P53蛋白高表达有关.

To investigated the anti-tumor in vivo effect and mechanism of the acid RNA protein complex (FA-2-b-β) of Agaricus blazei Murrill extract.CCK-8 method was used to detected the inhibitory effect of FA-2-b-β on proliferation of primary CML cells from newly diagnosed CML patients, the CML mouse model was established by trail-venous injection of primary CML cells, and the survival time, blood cell count and body weight were observed, the immunoflouresence and immunehistochemistry analysis, RT-qPCR, Western bolt were used to detemine the expression of caspase-3 signal pathway-related apoptosis genes and proteins.The experiments in vitro showed that the proliferative inhibitory rate in drug-treated group increased with concentration- and time-dependent manner (rThe FA-2-b-β can induce apoptosis of primary CML cells and prolong the survival time of CML model mouse, which may be related with the caspase-3 signal pathway related genes and proteins.FA-2-b-β对慢性髓系白血病原代细胞的增殖抑制作用及其相关机制.探讨秦巴硒菇提取物酸性RNA蛋白复合物（FA-2-b-β) 抗慢性髓系白血病的效果及机制。.应用CCK-8法检测不同浓度FA-2-b-β对初诊慢性髓系白血病患者原代细胞增殖抑制作用；通过尾静脉注射原代白血病细胞建立慢性髓系白血病小鼠模型, 观察模型小鼠生存期、血细胞数、体重变化；采用免疫荧光、免疫组织化学法、RT-qPCR及Western blot 法检测caspase3信号通路相关凋亡基因和蛋白的表达。.体外实验显示, 各药物浓度组中原代细胞增殖抑制率升高, 且呈浓度时间依赖性（rFA-2-b-β诱导慢性髓系白血病原代细胞凋亡, 可延长慢性髓系白血病模型小鼠的生存期, 其机制可能与caspase-3信号通路相关凋亡基因和蛋白有关。.

Tyrosine kinase inhibitor (TKI) has significantly improved the treatment of chronic myeloid leukemia (CML), however the resistance often resulted in treatment failure. Currently, it is known that the survival of CML cells can be affected by regulating autophagy, oxidative stress and mitochondrial metabolism, among which autophagy is an evolution-conserved catabolism process, and closely related to the pathogenesis of CML, thus playing a dual role in regulating the biological characteristics of cells. On the one hand, autophagy can promote the apoptosis of CML cells, and also can induce the drug resistance of CML cells on the other hand. In this review, the effect of autophagy on CML cells was summarzed briefly, so as to provide a useful idea to explore the combination of TKI with the autophagy inhibitor or inducer to overcome the resistance of CML to TKI.自噬影响慢性粒细胞白血病细胞生存的研究进展.酪氨酸激酶抑制剂（tyrosine kinase inhibitor, TKI）的应用, 给慢性粒细胞白血病（chronic myeloid leukemia, CML）的治疗带来了巨大的进展, 但患者耐药性的产生常导致治疗失败。目前已知可通过调控自噬、氧化应激、线粒体代谢等方式影响CML细胞的存活。其中, 自噬作为一种进化保守的分解代谢过程, 被认为与CML的发生、发展有密切关系, 同时在调节细胞生物学特性方面具有双重作用, 一方面可以促进CML细胞凋亡, 另一方面又可以诱导CML细胞产生耐药。本文将从自噬角度出发, 围绕自噬影响CML细胞发挥作用的最新研究进展作一简要综述, 以期为研究TKI联合自噬抑制剂或诱导剂克服CML对TKI耐药提供新思路.

To investigate the effect of GÖ6976 on the proliferation of chronic myeloid leukemia cells and its toxic effect on normal cells and mice, so as to provide experimental basis for the effectiveness and safety of its clinical application.Different concentrations of GÖ6976 were applied to the K562 cells, human peripheral blood mononuclear cells (PBMNC) and normal BaF3 cells, MTT assay was used to detect the effect on cell proliferation. BALB/C mice were used to investigate the toxicity in vivo. The general situation, body weight and the number of white blood cells in peripheral blood were monitored during administration, the blood collected from eyeballs before and after administration was used for biochemical examination, at the same time, the liver, kidney and femurs were examined pathologically.GÖ6976 could significantly inhibit the proliferation of K562 cells, inhibition effect increased with increasing dose (r=0.9623). However, there was no significant change in the inhibitory effect on PBMNC and BaF3 cells. The pathological examination of organs in each group showed no abnormal manifestations such as inflammatory infiltration, while the change rate of leukocyte count in peripheral blood of high dose group fluctuated greatly (P＜0.05), which might be related to the inhibition of intracellular protein kinase C, and no abnormality was observed in blood biochemical indexes. In the low dose group, there was no significant difference in peripheral blood leukocyte count, blood biochemical index and histopathology during administration drug as compared with the control group.GÖ6976 possesses a significant inhibitory effect on the proliferation of K562 cells, and the inhibitory effect increases with increasing dose. Long-term application of 5.0 μmol/L and below concentrations of GÖ6976 shows no obvious inhibitory effect on PBMNC, BaF3 cells. Long-term application of 10 mg/kg and below concentrations of GÖ6976 shows no obvious toxic effect on BALB/c mice.GÖ6976对慢性髓系白血病细胞增殖的影响及其毒性研究.研究GÖ6976对慢性髓系白血病（CML）细胞增殖的影响, 及其对正常细胞及小鼠的毒性, 为以后临床应用的有效性及安全性提供实验依据。.采用不同浓度的GÖ6976作用于CML急变期细胞株K562、人外周血单个核细胞（PBMNC）及小鼠正常BaF3细胞, MTT法检测其对细胞增殖的影响；选用BALB/C小鼠进行体内毒性研究, 给药期间监测一般情况、体重及外周血白细胞数；给药前后摘眼球取血进行生化检查, 同时对肝、肾及股骨进行病理学检查。.GÖ6976能明显抑制K562细胞的增殖, 抑制效果随着剂量的增加而增加（r=0.9623）, 但对PBMNC及BaF3细胞无明显抑制作用。各组脏器的病理学检查均显示无炎症浸润等异常表现；高剂量组在外周血白细胞数变化率波动较大（P＜0.05）, 这可能与细胞内蛋白激酶C被抑制有关。血液生化指标检测未见异常。低剂量组在外周血白细胞数、血液生化指标及组织病理学上与对照组比较无统计学差异。.GÖ6976对K562细胞增殖有明显抑制作用, 且抑制效果随着剂量增加而增加。长期应用GÖ6976 5.0 µmol/L及以下浓度对PBMNC、BaF3细胞没有明显的抑制作用。长期应用GÖ6976 10 mg/kg及以下浓度对BALB/c小鼠也没有明显的毒性作用。.

To report the clinical characteristics in a case of extramedullary T-lymphoblastic blast crisis of de novo chronic myelogenous leukemia (CML) so as to improve the understanding of this disease.The clinical characteristics, diagnostic approach and treatment of the patient were retrospectively analyzed, and some related literatures were reviewed.According to resuts of blood routine, bone marrow, chromosome and fusion gene tests, this patient was considered to be CML patient. The cervical lymph node biopsy indicated a T-cell lymphoblastic lymphoma (TLBL), and fluorescence in situ hybridization (FISH) analysis showed the BCR-ABL fusion gene within tumor cells of the patient's lymphnodes, thus was confirmedly diagnosed as extramedullary T-lymphoblastic blast crisis of chronic myelogenous leukemia. Treatment with dasatinib 140 mg/d combined with chemotherapy was then initiated, while the patient never achieved a complete remission.De novo chronic myelogenous leukemia in blast crisis is infrequent presence, the cases of extramedullary T-lymphoblastic blast crisis of newly diagnosed CML with additional chromosome 11q23 are extremely rare. And prognosis of these patients are poor, allogeneic hematopoietic stem cell transplantation maybe the only curable treatment.新诊断慢性髓系白血病髓外T淋巴细胞急变患者的临床特点分析.报道1例新诊断慢性髓性白血病（chronic myelogenous leukemia，CML）髓外T淋巴细胞急变的患者的临床特点，以提高对该疾病的认识.回顾性分析该患者的临床特征、诊疗方法，并对相关文献进行复习.结合血常规、骨髓学、染色体和融合基因检查结果，考虑该患者为CML，患者淋巴结病理提示为T淋巴母细胞淋巴瘤；荧光原位杂交检测发现淋巴结病变中存在BCR-ABL融合信号，最终诊断为CML髓外T淋巴细胞急变。给予达沙替尼（140 mg/d）联合化疗，患者一直未达完全缓解.新诊断的CML急变期患者相对少见，以CML髓外T淋巴细胞急变为特点并伴有附加染色体11q23的患者更为罕见。CML髓外T淋巴细胞急变的患者预后差，异基因造血干细胞移植或许是唯一有希望治愈的治疗方法.

To investigate the diagnosis and treatment of esophageal granulocytic sarcoma derived from chronic myelocytic leukemia (CML).The clinical manifestations, diagnosis and treatment of 1 case of esophageal granulocytic sarcoma secondary from chronic myelocytic leukemia were retrospectively analyzed and the related literature was reviewed.The patient was a 72-year-old woman with poststernal pain accompanied by general weakness. Gastroscopy was performed in a local hospital. At the same time, the increase of peripheral blood leucocytes was obvious. Under gastroscopy, 1.0 cm×0.5 cm irregular protuberance was found at 28 cm from the esophagus to the incisor teeth, and the surface was covered with erosion and a small amount of blood. Pathological results showed that heterotypic lymphoid cell infiltration, cytoplasmic red staining and more neutrophils were seen. Immunohistochemical staining results showed that AE1/AE3, CK5/6 and p63 displayed squamous epithelium (+); atypical lymphoid cells CD20Esophageal granulocytic sarcoma is rare in clinic, its clinical symptoms are not specific. Gastroscopy should be routinely screened for esophageal discomfort, and the esophageal granulocytic sarcoma derived from CML is treated according to the therapeutic regimen of the acute transformation of chronic myelocytic leukemia.继发于慢性髓系白血病的食道粒细胞肉瘤患者的临床分析.探讨食道粒细胞肉瘤的临床特点和诊治方法.对1例继发于慢性髓系白血病 (CML) 的食道粒细胞肉瘤的临床表现、诊断及治疗进行回顾性分析.老年女性72岁, 因胸骨后疼痛伴乏力, 在当地医院行胃镜发现食管距门齿28 cm处可见1.0 cm×0.5 cm欠规则隆起, 表面覆糜烂及少量鲜血；病理结果提示, 见异型的淋巴样细胞浸润, 胞质红染, 另见较多的中性粒细胞浸润; 同时发现外周血白细胞增高, 转诊我院。免疫组化标记结果显示, AE1/AE3、CK5/6与p63 显示鳞状上皮 (+) ；异型的淋巴样细胞CD20食道粒细胞肉瘤临床罕见, 临床症状无特异性, 对于白血病患者出现食道症状需要用胃镜进行筛查, 对于继发性慢性髓系白血病的食道粒细胞肉瘤需要按照慢性髓系白血病急性变进行治疗.

To investigate the effect of sphingosine-1-phosphate receptor 2 (S1PR2) specific antagonist JTE-013 on the proliferation of human chronic myeloid leukemia (CML) cell line K562.K562 cells were treated with JTE-013 (0, 0.5, 1, 5, 10, 20 μmol/L) for 24 and 48 hours respectively, CCK8 assay was used to detect the cell viability. K562 cells were treated with JTE-013 (0, 5, 10, 20 μmol/L) for 24 hours, propidium iodide (PI) DNA staining was used to analyze the cell cycle, Western blot was used to determine the levels of P21 and Cyclin D1 protein expression.JTE-013 inhibited the proliferation of CML cell line K562 in a dose dependent manner (r=-0.971). The proliferation rate of CML cells showed that the activity of CML cells decreased gradually with the increase of JTE-013 concentration (r=-0.971). The detection demonstrated that JTE-013 suppressed tumor cell proliferation through cell cycle arrest in GJTE-013, a S1PR2 antagonist, can inhibit the proliferation of human CML K562 cells, which may be achieved by arresting the cells in GS1PR2拮抗剂JTE-013抑制白血病细胞增殖.探讨鞘氨醇-1-磷酸受体2（sphingosine-1-phosphate receptor 2，S1PR2）特异性拮抗剂JTE-013对人慢性髓系白血病（CML）K562细胞增殖的影响.将JTE-013（0，0.5，1，5，10，20μmol/L）分别作用于人慢性髓系白血病细胞系K562细胞24、48 h，采用CCK-8法检测细胞活力；用JTE-013（0，5，10，20 μmol/L）处理K562细胞24 h，采用碘化丙啶（PI）DNA染色法检测细胞周期变化；采用Western blot检测细胞周期蛋白P21和Cyclin D1表达水平.JTE-013呈浓度依赖性抑制CML细胞系K562细胞增殖（r=-0.971）。实验结果显示，随着JTE-013作用浓度的升高，K562细胞活力逐渐降低（r=-0.971），且这种增殖抑制作用是通过将K562细胞阻滞在GS1PR2拮抗剂JTE-013可抑制人CML细胞系K562细胞增殖, 可能是通过将细胞阻滞在G

To investigate the curative efficacy of first generation TKI in the treatment of CML-CP combined with vPh and genetic characteristics.60 patients with CML-CP combined with vPh from January 2010 to May 2017 in the First Affiliated Hospital of Kunming Medical University were chosen as CML-CP-vPh group, and 107 patients with CML-CP combined with typical Ph chromosome at the same time were chosen as control group. The patients in two groups were treated with imatinib; The curative efficacy, karyotype and FISH signal type were compared between 2 groups, and the factors influencing long-term survival of patients were analyzed by Cox risk model.There was no significant difference in the demographic and hematological baseline data between 2 groups (P＞0.05), and there was no significant difference in the incidence of drug-resistance between 2 groups (P＞0.05). The incidence of primary drug-resistance and primary hematological drug-resistance in the CML-CP-vPh group were significantly higher than that in control group (P＜0.05). There was no significant difference in the accumulative CCyR rate, accumulative MMR rate, OS and EFS between 2 groups (P＞0.05). Multivariate Cox model analysis showed that vPh not correlated with OS and EFS of patients with CML-CP (P＞0.05). The factors influencing OS in CML-vPh patients included high risk of Sokal scores, peripheral blood basophils proportion ≥10%, BCR-ABLPatients with CML-CP combined with vPh possess higher primary drug-resistance rate and primary hematological drug-resistance rate for the first generation TKI, while second-generation TKI can efficiently improve long-term survival, its efficacy is similar to efficacy for patients with typical Ph chromosomes. CML-CP combined with vPh does not display special demographic and hematological characteristics. The main involved regions of heterotopic variants include 12q1, 12q2 and 1p3, while 2G2R1Y type is the most common type of FISH signal.第一代TKI治疗初治CML-CP合并vPh患者的疗效及遗传学特征.探讨第一代酪氨酸激酶抑制剂（TKI）治疗初治慢性髓系白血病慢性期（CML-CP）合并Ph染色体变异易位（vPh）的疗效及遗传学特征.选取昆明医科大学第一附属医院2010年1月到2017年5月收治的初治CML-CP合并vPh患者共60例，设为CML-CP-vPh组，以同期初治CML-CP合并典型Ph染色体患者107例设为对照组，2组均给予伊马替尼治疗。比较2组临床疗效、染色体核型特征及FISH信号类型特征，并采用Cox风险模型进行患者远期生存影响因素分析.2组人口学和血液学基线资料比较差异无显著性（P＞0.05）；2组耐药发生率比较差异无显著性（P＞0.05）；但CML-CP-vPh组原发耐药和原发血液学耐药发生率均显著高于对照组（P＜0.05）。2组累积CCyR率、累积MMR率、OS及EFS比较差异无显著性（P＞0.05）。多因素Cox模型分析提示，vPh与CML-CP患者的OS和EFS并无相关性（P＞0.05）； CML-vPh患者OS影响因素为Sokal评分高危、外周血BBP比例≥10%、BCR-ABLCML-CP合并vPh患者具有更高的对第一代TKI原发耐药率及原发血液学耐药率，而改用第二代TKI可改善患者长期生存，与合并典型Ph染色体者疗效接近。CML-CP合并vPh患者并无特殊人口学和血液学特征，变异异位多累及12q1、12q2及1p3区域，而FISH信号类型则以2G2R1Y型最为常见.

To investigate the effects of CPEB4 on the migration and cycle of K562 cells and the changes of protein molecules that may be involved in the regulatory mechanism.Western blot was used to detect the expression of CPEB4 in normal leukocytes and K562 cells. The overexpression plasmid pcDNA3.1(+)-His-CPEB4, silencing plasmid pPLK+Puro-CPEB4 shRNA were transfected into K562 cells by electroporation so as to change CPEB4. The transfection efficiency was detected by Western blot. Finally, the migration and cycle of different cells were detected by Transwell chamber and flow cytometry.Western blot was used to detect the expression changes of MMP2, MMP9, CDK4, CyclinD1 and P21 proteins.Compared with normal white blood cells, the expression of CPEB4 protein in K562 cells was significantly enhanced (P＜0.01); Compared with the control group, CPEB4-silenced K562 cells showed that the cell migration ability was significantly enhanced (P＜0.01); GCPEB4 can inhibit the migration of K562 cells and arrest cell cycle progression at GCPEB4对慢性髓系白血病细胞迁移与周期的影响.探讨CPEB4对慢性髓系白血病细胞K562迁移与周期的影响及可能参与机制调控的蛋白分子的变化.采用Western blot检测正常白细胞和K562细胞中CPEB4的表达水平；再将过表达质粒pcDNA3.1(+)-His-CPEB4、沉默质粒pPLK+Puro-CPEB4 shRNA电穿孔转染K562细胞，改变CPEB4在K562细胞中的表达量，利用Western blot检测转染效率；最后利用Transwell小室、流式细胞术检测不同处理细胞的迁移、周期情况，并用Western blot检测MMP2、MMP9、CDK4、CyclinD1、P21蛋白的表达变化.与正常白细胞相比，K562细胞中的CPEB4蛋白表达量明显升高（P＜0.01）；CPEB4沉默的K562细胞与对照组相比，细胞的迁移能力显著提高（P＜0.01）；GCPEB4可抑制K562细胞迁移，将细胞周期进程阻滞于G

To analyze the significance of various abnormal signal patterns appreared in CML and B-ALL patients by using BCR/ABL/ASS1 tricolor dual-fusion probe, and to explore its application value in detecting BCR/ABL fusion gene and ASS1 gene deletion.50 newly diagnosed CML patients and 50 newly diagnosed B-ALL patients were detected by fluorescence in situ hybridization (FISH) with BCR/ABL/ASS1 tricolor dual-fusion probe. Meanwhile, karyotype analysis was performed on all the patients using the 24 hours short-term culture and R-banding.Among the 50 CML patients, PhTricolor dual-fusion FISH probe for detecting BCR/ABL fusion gene and ASS1 gene deletion is simple, rapid, sensitive and stable. It can detect various forms of molecular fusion and avoid the false positive results due to coincidental overlap of signals generated by D-FISH probe and ES-FISH probe. In addition, this detection method not only can directly observe the presence or absence of ASS1 gene deletion, but also improve the reliability of the positive results of newly diagnosed BCR/ABL fusion gene and accuracy of monitoring results of minimal residual disease for the subsequent visit.应用三色双融合探针检测BCR-ABL融合基因及ASS1基因缺失.分析BCR/ABL/ASS1三色双融合探针在CML和B-ALL所出现的各种异常信号模式的意义，并探讨其在检测BCR/ABL融合基因及ASS1基因缺失中的应用价值.分别对50例初诊CML患者和50例初诊B-ALL患者采用BCR/ABL/ASS1三色双融合探针进行荧光原位杂交（FISH）检测，同时对所有病例应用24 h短期培养法R显带后进行染色体核型分析.50例CML患者中，49例Ph应用三色双融合FISH探针检测BCR/ABL融合基因及ASS1基因缺失，具有简单、快速、灵敏、稳定的特点，能够检测多种形式的分子融合，可很好地避免D-FISH探针及ES-FISH探针因信号随机重叠导致的假阳性结果。该检测方法不但可以直接观察有无ASS1基因的缺失，而且还能提高初诊BCR/ABL融合基因阳性结果的可靠性及复诊监测微小残留病结果的准确性.

To investigate the effect and mechanism of miR-124-3p-targeing regulating ABCA2 on chronic myelogenous leukemia cell K562-R.CML cells with miR-124-3p-overexpression and ABCA2-over-expression as well as subcutaneoustrans planted tumor nude mice were used as study objects. And the CML cells were divided into four groups: K562-R blank control, miR-124-3p mimic control, ABCA2-overexpression and mimic+PC ABCA2. The effects of miR-124-3p and ABCA2 on CML cells were analyzed. The levels of proliferation-, apoptosis- and autophagy- related protein were determined by Western blot. qRT-PCR was employed to detect the levels of miR-124-3p and ABCA2 in K562-R cells. The relationship between miR-124-3p and ABCA2 was validated by luciferase reporter system assays and bioinformatics. Hoechst/immunohistochemical staining and CCK-8 assay were performed to investigate the function involved.miR-124-3p highly expressed in K562-S cells and lowly expressed in K562-R cells, however, ABCA2 lowly expressed in K562-S cells and highly expressed in K562-R cells. Over-expression of miR-124-3p significantly decreased ABCA2 level and cell growth, but increased autophagy and apoptosis in K562-R cells (P＜0.01). When ABCA2 was over-expressed, the K562-R cell growth was promoted and autophagy and apoptosis were inhibited (P＜0.01). The miR-124-3p promoted cell autophagy and apoptosis but inhibited cell growth in nude mice transplant tumor model (P＜0.01).miR-124-3p can target ABCA2 to inhibit the growth of CML cells and promote the cell autophagy and apoptosis of CML cells.miR-124-3p靶向ABCA2增强慢性白血病细胞K562-R对伊马替尼的敏感性.探讨miR-124-3p靶向ABCA2对CML细胞株耐药性及细胞活性的影响.过表达miR-124-3p和ABCA2的CML细胞株K562-R及裸鼠皮下移植瘤模型用于本研究。设置K562-R空白对照组、miR-124-3p mimic对照组、ABCA2过表达组和mimic+pc ABCA2组，研究miR-124-3p和ABCA2对K562-R细胞的影响。Western Blot检测体系中增殖、凋亡和自噬相关分子表达；qRT-PCR检测体系中miR-124-3p和ABCA2的表达；荧光素酶系统及生物信息学软件预测miR-124-3p和ABCA2之间靶向关系。CCK-8染色检测细胞增殖，Hoechst染色检测细胞凋亡，免疫组织化学检测细胞凋亡和增殖相关分子.miR-124-3p在K562耐药细胞（K562-R）中低表达，在K562敏感细胞（K562-S）中高表达；ABCA2在K562-R中高表达，在K562-S中低表达（P＜0.01）；生物信息学和荧光素酶报告系统检测显示，miR-124-3p靶向调节ABCA2表达。miR-124-3p过表达能有效抑制K562-R细胞增殖，促进细胞凋亡和自噬发生（P＜0.01）；而ABCA2过表达则促进K562-R细胞增殖，明显抑制凋亡和自噬发生（P＜0.01）；进一步通过裸鼠体内皮下移植瘤模型表明，miR-124-3p可以有效抑制K562-R细胞增殖，促进细胞凋亡和自噬（P＜0.01）.miR-124-3p可以通过靶向ABCA2来有效抑制白血病耐药细胞株K562-R的增殖，促进细胞凋亡和自噬.

Chronic myeloid leukemia (CML) is a malignant myeloproliferative tumor which is originating from hematopoietic stem cells. Chemotherapy is the preferred made of treatment for the disease. However, in recent years, more and more patients have multidrug resistance (MDR) during treatment, which is the main cause of failure treatment, therefore, the search for effective reversal agents has important clinical significance. Traditional Chinese medicine derived reversal agents have the characteristics of multiple targets, low toxicity and high efficiency, which can reverse the drug resistance through different mechanisms, and the research potential is unlimited. In recent years, it has been widely concerned by scholars at home and abroad, especially the research on physcion, emodin, curcumin, matrine, gambogic acid, oridonin, ligustrazine, solanine and other monomers, which has made great advance. In this reviwe, the recent research advance on the reversal of MDR in chronic myeloid leukemia by Chinese medicine monomer is summarized briefly.中药单体逆转慢性髓系白血病多药耐药的研究进展.摘要 慢性髓系白血病（CML）是一种起源于造血干细胞的恶性骨髓增生性肿瘤。化疗是该病首选的治疗方式，但近年来越来越多的患者在治疗中出现多药耐药（multidrug resistance，MDR），这是导致治疗失败的主要原因，因此寻找有效逆转剂对临床具有重要的意义。中药逆转剂具有多靶点、低毒、高效等特点，其可以通过不同的机制逆转耐药，具有无限的研究潜能，近几年受到了国内外学者的广泛关注，特别是对大黄素甲醚、大黄素、姜黄素、苦参碱、藤黄酸、冬凌草甲素、川芎嗪、龙葵碱等单体的研究有不少进展。本文就近年来中药单体逆转慢性髓系白血病MDR的最新研究进展作一综述.

To investigate the function and mechanism of transcription factor of MEIS1 and miR-425 to the proliferation of chronic myeloid leukemia cell K562.Bioinformatic prediction was used to analyze the binding of MEIS1 in miR-425 promoter region. ChIP-qPCR coupled with dual luciferase assay was used to detect the combination of MEIS1 and the transcription activity of miR-425, and its regulative role in the transcription activity miR-425. CCK-8 was used to detect the effect of MEIS1 and miR-425 on cell proliferation. Flow cytometry with PI staining was used to detected the effect of MEIS1 and miR-425 on K562 cell cycle progression. Western blot was used to examine the effect of miR-452 on the expression level of MEIS1.MEIS1 could bind the promoter of miR-425 and repressed its transcription. After K562 was transfected by shRNA, the K562 cell proliferation and cell cycle progression was significantly inhibitied. Moreover, after K562 cells were transfected by miR-425 mimic, cell proliferation and cell cycle was inhibited. The expression level of MEIS1 could be inhibited by the combination of miR-425 and MEIS1 3'UTR.MEIS1 can inhibit the activity of miR-425 in transcriptional level, while the miR-425 can suppress the expression of MEIS1 protein in post-transnational level. Therefore, a regulatory circuit comprising from MEIS1 and miR-425 regulates K562 cell proliferation.MEIS1/miR-425反馈调节通路对慢性髓系白血病细胞株K562增殖的作用.研究转录因子MEIS1和miR-425对人慢性髓系白血病细胞株K562增殖的调控作用及其分子机制.通过生物信息学预测分析转录因子MEIS1在miR-425启动子区的结合位点情况，ChIP-qPCR结合双荧光报告基因实验检测MEIS1与miR-425启动子的结合情况及其对miR-425启动子的调节作用，CCK-8法检测MEIS1和miR-425对K562细胞增殖的影响，PI染色结合流式细胞术检测MEIS1和miR-425对K562细胞周期的影响，Western blot检测miR-425对MEIS1表达的影响.MEIS1可以与miR-425启动子区结合，并可抑制其活性。使用针对MEIS1的shRNA转染K562细胞后，可显著抑制K562细胞增殖及细胞周期的运行。类似地，使用miR-425的模拟物转染K562细胞后，也可抑制K562细胞的增殖和细胞周期的运行。miR-425可以通过与MEIS1 3′UTR的结合抑制MEIS1的表达.转录因子MEIS1可在转录水平抑制miR-425的转录活性，miR-425则在转录后水平抑制MEIS1蛋白的表达，因此，两者共同构成一个反馈调节通路作用于K562细胞的增殖.

To investigate the relationship between long non-coding RNA (LncRNA) PANTR1 and imatinib resistance in chronic myeloid leukemia cell line K562 and its mechanism.K562 control cells (Control) and K562 imatinib resistant cells (ImR) were cultured. Two siRNA vectors targeting PANTR1 and control vectors were transfected into K562-ImR cells by lentivirus as ImR-siPA#1, ImR-siPA#2 and ImR-siControl cells, respectively. Imatinib semi-inhibitory concentration (ICImatinib ICLncRNA PANTR1 can promote the expression of MDR and stem cell marker in chronic myeloid leukemia cell line K562, and mediate imatinib resistance.PANTR1与慢性髓系白血病细胞K562对伊马替尼耐药的关系及其相关机制.探讨长链非编码RNA（LncRNA）PANTR1与慢性髓性白血病细胞K562对伊马替尼耐药的关系及机制.培养K562对照细胞（Control）及K562伊马替尼耐药细胞（ImR），采用慢病毒转染靶向PANTR1的2种siRNA载体及对照载体于K562-ImR细胞中，分别为ImR-siPA#1、ImR-siPA#2及ImR-siControl细胞；采用CCK-8实验检测各组细胞伊马替尼半抑制浓度（ICImR细胞伊马替尼ICLncRNA PANTR1可促进慢性髓系白血病细胞K562中MDR的表达及其干细胞标志物的表达，介导细胞对伊马替尼耐药.

To investigate the biological effects and mechanism of WNK1 on K562 cells through regulating MAPK7.Cells were routinely cultured in vitro, the expression of WNK1 and MAPK7 in different blood tumor cell lines was analyzed by RT-qPCR and Western blot analysis.K562 cells were transfected with siRNA-WNK1 lentivirus.The effect of WNK1 on K562 cell proliferation was analyzed by using CCK-8 reagent.And K562 cell apoptosis was analyzed by using flow cytometry. The expression level of phosphorylated MAPK7 protein and total MAPK7 protein in K562 cells was analyzed by Western blot.The mRNA and protein of WNK1 were highly expressed in HL60, THP-1, U266 and K562 cells, however, the expressions were the highest in K562 cells (P0.05), while the changes of mRNA and protein expressions of MAPK7 were not significantly in HL60, THP-1, U266 and K562 cells (P0.05), but the phosphorylated MAPK7 expression was the highest in K562 cells (P0.05). Proliferation of K562 cells transfected by WNK siRNA was significantly suppressed, and apoptosis was obviously increased (P0.05). And the pMAPK7 protein expression in K562 cells transfected by WNK1 siRNA significantly decreased (P0.05), however, the total MAPK7 protein expression in K562 cells showed no obvious change (P0.05).WNK1 is highly expressed in K562 cells, which can promote the proliferation of K562 cells and reduce apoptosis probably by promoting phosphorylation of its downstream MAPK7.WNK1通过磷酸化激活MAPK7对人慢性髓系白血病K562细胞的影响及相关机制.探讨WNK1通过调控MAPK7对人慢性髓系白血病K562细胞的影响及相关机制.通过体外培养不同肿瘤细胞，采用RT-qPCR和Western blot分析WNK1、MAPK7在不同血液肿瘤细胞系中的表达。通过慢病毒转染siRNA-WNK1至K562细胞，采用流式细胞术分析细胞凋亡情况，以及K562细胞中总MAPK7、磷酸化MAPK7表达的变化，通过CCK-8方法检测细胞增殖，并进行统计学分析.WNK1的mRNA及蛋白在HL-60、THP-1、U266和K562细胞中均高表达，但是在K562细胞中表达水平最高（P＜0.05），MAPK7的mRNA及总蛋白表达在HL60、THP-1、U266和K562细胞中均无明显变化（P0.05），但是在K562细胞中磷酸化MAPK7的表达最高（P＜0.05）。转染WNK1 siRNA的K562细胞增殖受到明显抑制、细胞凋亡增加（P＜0.05）。转染WNK1 siRNA的K562细胞磷酸化MAPK7蛋白表达显著下降（P＜0.05），但是总MAPK7蛋白无明显变化（P0.05）.WNK1在K562细胞中高表达，能够促进K562细胞增殖、抑制凋亡等恶性细胞生物学行为，其机制可能是促进了下游MAPK7的磷酸化.

Chronic myelogenous leukemia (CML) is a hematological malignancy that seriously threatens the lives of patients. It was found that there are abnormal classic Wnt pathway, that is, Wnt/β-catenin signaling pathways in CML cells, moreover, Wnt/β-catenin signaling pathway is involved in the growth and proliferation of CML cells, and closely relates with the self-renewal ability of CML leukemic stem cells. This review summarizes the recent studies on the relationship between Wnt/β-catenin signaling pathway and CML, and the researches on the targeting inhibition of Wnt/β-catenin signaling pathway in CML treatment, thus to provide new ideas for the treatment of CML.经典Wnt途径在慢性髓系白血病中的研究进展.慢性髓系白血病（chronic myelogenous leukemia，CML）是一种严重威胁患者生命的血液系统恶性疾病。在CML细胞中存在着经典Wnt途径即Wnt/β-catenin信号通路的异常，且该信号通路参与了CML细胞生长、增殖等生命活动，与CML白血病干细胞的自我更新能力密切相关。本文基于近几年的文献对Wnt/β-catenin信号通路与CML之间的关系，以及靶向抑制Wnt/β-catenin信号通路治疗CML的最新研究进行综述，为CML的治疗提供新思路.

To analyze the relationship between cytogenetic changes and the progression in the patients with chronic myeloid leukemia (CML) during the treatment with tyrosine kinase inhibitor (TKI).The chromosome G banding of 150 patients with CML treated in our hospital, was carried out to analyze the karyotype by the 24 h short-term culture or direct method of bone marrow cells, and the point mutation of the ABL kinase area was detected, the relationship between cytogenetic changes and the evolution of the disease course was analyzed.The indirect fluorescence in situ hybridization showed that the BCR-ABL fusion gene of 150 patients was positive, out of which 142 cases showed positive Philadelphia (Ph) (94.67%), 8 cases with Ph negative (5.33%). Among 142 cases with Ph positive on the first diagnosis, and 14 cases (9.86%) with additional chromosome abnormality (9.86%), 4 cases (2.82%) with mutation translocation with 124 cases (87.32%), standard translocation t (9; 22) (q34; Q11) were found. Out of the 14 patients with additional chromosomal abnormalities, 8 cases with "main pathmay" abnormalities, 2 case with -Y abnormalities, and 4 cases with "secondary pathway" abnormalities were observed. During TKI treatment, additional chromosomal abnormalities were found in 46 patients with standard translocation and abnormal number of chromosomes, and the incidence of disease progression and point mutation were higher (P＜0.05). Compared with patients with the standard translocation, the disease-free survival rate of the patients diagnosed as CML at 1st visit and with additional chromosome abnormality was significantly decreased (P＜0.05), but the overall survival rate showed no significantly different (P＞0.05). Compared with patients without additional cvtogenetic aberrations, the disease free and overall survival rate of the patients with additional cytogenetic aberrations during the TKI treatment of CML in chronic phase were significantly decreased (P＜0.05).Some CML patients may have additional chromosomal abnormalities during the onset and development of the disease, and these patients are at higher risk of disease progression.CML患者在TKI治疗过程中的细胞遗传学变化与病程演进的关系分析.分析慢性髓系白血病（CML）患者在酪氨酸激酶抑制剂（TKI）治疗过程中的细胞遗传学变化与病程演进的关系.对本院收治的150例初诊CML患者，通过骨髓细胞24 h短期培养法或直接法，行染色体G显带技术核型分析；经ABL激酶区点突变检测，分析细胞遗传学变化与其病程演进的相关性.间接荧光原位杂交技术检测结果发现，150例患者BCR-ABL融合基因均为阳性，其中费城（Ph）染色体阳性142例（94.67%），阴性8例（5.33%）；142例Ph染色体阳性患者中，初诊Ph阳性且伴有额外染色体异常14例（9.86%），变异易位4例（2.82%），标准易位t（9；22）（q34；q11）124例（87.32%）。14例伴有额外染色体异常的患者中，“主要路径”异常8例，-Y异常2例，“次要路径”异常4例。在TKI治疗期间，46例标准易位患者出现额外染色体异常，以染色体数目异常为主，此类患者疾病进展和出现点突变的比例较高（P＜0.05）。与标准易位患者比较，初诊CML慢性期伴有额外染色体异常患者无病生存率明显降低（P＜0.05），而与总生存率的比较，并无明显差异（P＞0.05）。与无额外染色体异常患者比较，CML慢性期TKI治疗期间出现额外染色体异常者无病生存率及总生存率明显下降（P＜0.05）.部分CML患者在疾病发生和发展过程中可出现额外染色体异常，此类患者发生疾病进展的风险性较高.

To explore whether BAX plays a role in the development of Philadelphia chromosome-positive leukemia and related mechanisms.Target-gene knockout mice were used as bone marrow cell donors. Retrovirus over-expressing BCR-ABL were packaged. BCR-ABL-induced B-ALL mouse model was established through donor's B cells transfected by the retrovirus and the B cells over-expressing BCR-ABL were given to the receptor mice by tail vein injection. Western blot was used to detect the protein express and flow cytometry was used to analyze the B cell subpopulations in BAXBAX deletion caused faster development of BCR-ABL-induced leukemia in vitro and in vivo. BCR-ABL increased BCL-2 expression and enhanced BCL-2/BAX heterodimer formation.The BAX deletion can accelerate the disease progression of BCR-ABL induced B-ALL.BAX基因缺失加速BCR-ABL诱导的小鼠B-ALL疾病进程.探讨BAX对BCR-ABL诱导的小鼠B-ALL的发生、发展的影响及相关机制.以目的基因敲除小鼠作为骨髓细胞供体；包装过表达BCR-ABL的逆转录病毒；通过逆转录病毒感染供体B-细胞和将过表达BCR-ABL的B-细胞尾静脉注射给受体小鼠，构建BCR-ABL诱导的B-ALL小鼠模型。采用Western blot检测蛋白的表达，采用流式细胞术分析细胞，用Kaplan-Meier曲线分析发病小鼠生存期.BCR-ABL转染的BAX缺失B-细胞比野生型B-细胞在体外和受体小鼠体内增长更快；BAX缺失供体B-细胞构建的疾病小鼠的生存期更短。BCR-ABL增加BCL-2表达和BCL-2/BAX异二聚体形成.BAX缺失会加速BCR-ABL诱导的小鼠B-ALL疾病进程.

To explore the expression of CPEB4 in K562 cells, the biological activity and its possible molecular mechanisms.Western blot was used to detect the expression of CPEB4 in normal leukocytes and K562 cells. The overexpression plasmid pcDNA3.1(+)-His-CPEB4 and the silencing plasmid pPLK+Puro-CPEB4 shRNA were transfected into K562 cells to change the expression of CPEB4 in K562 cells, and the transfection efficiency was detected by Western blot. Finally, CCK-8 and flow cytometry were used to detect the proliferation and apoptosis of differently treated cells, and the expression changes of proliferation and apoptosis marker proteins (AKT, p-AKT, caspase-3, BCL-2) were detected by Western blot.Compared with normal leukocytes, the expression of CPEB4 protein in K562 cells was higher (P＜0.01). Compared with the control group, the proliferation of CPEB4-silenced K562 cells significantly increased (P＜0.01), the number of apoptotic cell significantly decreased, and expression of AKT, p-AKT and BCL-2 was significantly increased, the protein expression of caspase-3 was significantly reduced. The proliferation of K562 cells after CPEB4 overexpression was slowed down (P＜0.05), the number of apoptotic cells significantly increased,the expressions of AKT, p-AKT and BCL-2 were significantly down-regulated, and the expression of caspase-3 was up-regulated.CPEB4 can inhibit the proliferation and promote the apoptosis of K562 cells, the AKT, p-AKT, BCL-2 and caspase-3 are involved in the regulation mechanism.CPEB4对慢性髓系白血病细胞增殖与凋亡的影响.探索CPEB4在K562细胞中的表达、生物学活性及其可能的分子机制.采用Western blot检测正常白细胞和K562细胞中CPEB4的表达情况；再将过表达质粒pcDNA3.1（+）-His-CPEB4、沉默质粒pPLK+Puro-CPEB4 shRNA电穿孔转染K562细胞，改变CPEB4在K562细胞中的表达量，应用Western blot检测转染效率；最后应用CCK-8和流式细胞术检测不同处理的细胞增殖和凋亡情况，并用Western blot法检测增殖与凋亡标志蛋白（AKT、p-AKT、caspase-3、BCL-2）的表达变化。.与正常白细胞相比，K562细胞中CPEB4蛋白表达量较高（P＜0.01）；CPEB4沉默的K562细胞与对照组相比，细胞增殖水平显著增高（P＜0.001），细胞凋亡数显著减少，AKT、p-AKT、BCL-2蛋白表达水平明显增高，而caspase-3蛋白表达显著降低；CPEB4过表达后的K562细胞增殖减慢（P＜0.05），细胞凋亡数明显增加，AKT、p-AKT和BCL-2蛋白表达显著降低，caspase-3蛋白表达增高。.CPEB4可抑制K562细胞增殖，促进K562细胞凋亡，而AKT、p-AKT、BCL-2和Caspase-3等分子参与调控机制。.

To investigate the effect of Quercetin (Qu) on cell proliferation, apoptosis and cell cycle, as well as the expression changes of Wnt/β-catenin signaling pathway, apoptosis and cell cycle regulators and BCR-ABL in CML susceptible cells K562 and imatinib-resistant cells (IM) K562R.The trypan blue staining was used to detect the all proliferation. The cell cycle and apoptosis were detected by flow cytometry. The fluorescence quantitative PCR and Western blot were used to detect the expression of mRNA and protein respectively.After administration with 5, 10, 20, 40, 80, 160, 320 μmol/L Qu, the inhibition ratio in K562 cells was 5.07%, 5.98%, 11.09%, 31.88%, 56.89%, 70.44%, 86.63%; and that in K562R cells were 4.99%, 9.75%, 10.54%, 8.93%, 25.13%, 46.89%, 68.60%; ICQu can inhibit the proliferation K562 and K562R cells, and decrease the drug resistance and increase the sensitivity, that relate with inhibiting Wnt/β-catenin signaling pathway, activating apoptosis pathway and cyclins.槲皮素对K562和K562R细胞Wnt/β-catenin信号通路的影响.观察槲皮素（Qu）对伊马替尼（IM）敏感细胞K562及耐药细胞K562R的增殖、凋亡、细胞周期的影响，探讨凋亡及细胞周期调控因子、Wnt/β-catenin信号通路及BCR-ABL的表达变化，为Qu的临床应用提供实验室依据.应用台盼蓝染色法检测K562和K562R细胞的增殖水平；流式细胞术检测细胞凋亡及细胞周期的分布；荧光定量PCR及Western blot法分别检测Wnt/β-catenin信号通路成员、细胞凋亡及细胞周期相关因子的mRNA及蛋白的表达.经Qu 5、10、20、40、80、160、320 μmol/L作用于K562细胞后，K562细胞抑制率分别为5.07%、5.98%、11.09%、31.88%、56.89%、70.44%、86.63%；K562R细胞抑制率分别为4.99%、9.75%、10.54%、8.93%、25.13%、46.89%、68.60%。K562、K562R的ICQu能够抑制K562及K562R细胞的增殖，降低其耐药性，增加敏感性，这与Qu抑制Wnt/β-catenin信号通路、激活凋亡途径及细胞周期因子相关.

To construct a K562 and adriamycin-resistant K562 (KAR) cell line with stably down-regulation of NCL (nucleolin) expression, and to investigate the effect of NCL down-regulation on the drug resistance in K562 and KAR cells.K562 and KAR cells were infected with lentivirus, and stably transfected cell clones were obtained by puromycin screening. The cell proliferation was detected by MTS assay, the cell apoptosis was detected by flow cytometry, and the expression level of drug resistance related genes was detected by real-time PCR.The K562 and KAR cells with stable down-regulation of NCL were successfully constructed. Compared with the control group, the proliferation of K562 and KAR cells with down-regulating NCL expression decreased significantly (P ＜0.05), the apoptosis of cells increased significantly (P ＜0.05), and cell resistance to adriamycin was down-regulated.Inhibition of NCL expression may increase the sensitivity of cells to adriamycin, which may be related with the promotion of apoptosis of K562 and KAR cells.抑制NCL表达对慢性髓系白血病细胞K562及其耐药细胞的影响.建立稳定下调核仁素（NCL）表达的K562细胞及耐阿霉素的K562细胞（KAR），研究下调NCL对K562及KAR细胞耐药作用的影响.用慢病毒感染K562及KAR细胞，经嘌呤霉素筛选获得稳定转染的细胞克隆。采用MTS法检测各组细胞的增殖能力，流式细胞术检测细胞凋亡情况，Real-time PCR检测耐药相关基因的表达水平.成功的构建了NCL表达下调的K562及KAR稳定转染细胞株。与对照组相比较，下调NCL的表达可以使细胞的增殖明显受抑（P＜0.05），细胞的凋亡显著增加（P＜0.05），细胞对阿霉素的耐药性降低.抑制NCL的表达可以提高细胞对阿霉素的敏感性，这一作用可能与其促进细胞凋亡有关.

At present, allogeneic hematopoietic stem cell transplantation is still the only way to cure chronic myelogenous leukemia. With the advances of HLA matching technology, application of tyrosine kinase inhibitors before and after transplantation, improvement of postoperative immune status and fusion gene monitoring, and the control of postoperative complications, especially graft-versus-host disease etc. allogeneic hematopoietic stem cell transplantation is displaying better efficacy in the treatment of chronic myelogenous leukemia, and the quality of life of patients has also been significantly improved. This article reviews the recent research advances on the allogeneic hematopoietic stem cell transplantation and related support technologies for treatment of chronic myeloid leukemia.慢性髓系白血病异基因造血干细胞移植的研究进展.目前，异基因造血干细胞移植（allogeneic hematopoietic stem-cell transplantation，allo-HSCT）依旧是根治慢性髓系白血病（chronic myelogenous leukemia，CML）的唯一方法。随着HLA基因配型技术的改进、移植前后酪氨酸激酶抑制剂（tyrosine kinase inhibitor，TKI）的应用、术后免疫状态及融合基因监测的进步、术后并发症尤其是移植物抗宿主病（graft-versus-host disease，GVHD）的控制得到加强等，异基因造血干细胞移植在慢性粒细胞白血病治疗中正展现出更好的疗效，患者的生存质量也有了显著提高。本文将就近年来异基因造血干细胞移植及其相关支持技术在慢性髓系白血病治疗中最新研究进展作一综述。.

To retrospectively analyze the clinical manifestation, laboratorial test features and prognosis of patients with CML in myeloid blast crisis.The clinical data of 10 patients with CML in myeloid blast crisis admitted in Chinese PLA General Hospital from June 2011 to May 2018 were collected, and their clinical features, laboratorial data and long-term survival were analyzed.The median age of these 10 cases was 32.5 (23-73) years old. Nine cases had chronic phase history. The median chronic phase was 17(4-84) months. All the 10 cases had splenomegaly; B-ultrasonography showed that the median spleen size was 5.2 (4-7.8) cm in thickness, and 14.6 (11.4-19.8) in length. When chronic myeloid leukemia was in blast crisis, the median WBC count was 41.705（11.9-344.41）×10CML patients in myeloid blast crisis treated by chemotheraphy combined with TKI gain CP2, the survival time of patients treated by sequential allo-HSCT is prolonged.慢性髓系白血病急髓变临床回顾性分析.单中心回顾性分析慢性髓系白血病急髓变患者的临床表现、实验室检查特征及预后.收集解放军总医院2011年6月至2018年5月间10例慢性髓系白血病急髓变患者临床资料，分析临床特征，实验室检查及长期生存情况.10例慢性髓系白血病急髓变患者中位年龄32.5（23-73）岁；9例有慢性病史，中位慢性期17（4-84）个月。10例患者均有脾脏肿大，B超结果显示中位脾厚5.2（4-7.8）cm，中位脾长14.6（11.4-19.8）cm；慢性髓系白血病急髓变时中位外周血白细胞数41.705（11.9-344.41）×10慢性髓系白血病急髓变患者行联合TKI化疗诱导达CP2状态，序贯异基因造血干细胞移植可延长患者生存期.

To investigate the role of nucleophosmin (NPM) in the proliferation of chronic myeloid leukemia cells (K562 cells) and its mechanism by RNAi technology.shRNA was used to inhibit the expression of NPM. The expression of NPM gene was detected by real-time quantitative PCR. The effect of inhibiting NPM gene on cell proliferation was detected by MTS assay. Change of cell cycle was detected by flow cytometry. Western blot was used to detect the expression of cell cycle-related proteins.The shRNA lentiviral vector targeting at NPM gene was successfully constructed and used to transfect the K562 cells. The results showed that compared with the control groups, suppression of NPM gene expression in K562 cells could inhibit the cell proliferation and decrease the cell colony formation. Moreover, interference of NPM gene could prolong GDown-regulation of NPM gene expression in K562 cells can induce cell cycle arrest and inhibit cell proliferation.下调NPM基因表达对慢性髓系白血病细胞株K562增殖的抑制作用研究.运用RNA干扰技术抑制核仁磷酸蛋白（NPM）的表达，研究NPM基因在慢性髓系白血病细胞株（K562细胞）增殖中的作用及其机制.采用shRNA抑制NPM的表达，应用实时荧光定量PCR技术检测NPM基因的表达，MTS法检测抑制NPM基因对细胞增殖能力的影响，流式细胞术检测细胞周期的改变，Western blot检测细胞周期相关蛋白的表达.成功构建了针对NPM基因的shRNA慢病毒载体，并感染K562细胞。检测结果显示，与对照组相比较，抑制K562细胞NPM基因的表达可以抑制细胞的增殖，减少细胞集落形成。干扰NPM基因可使G下调K562细胞NPM基因的表达，可以通过诱导细胞周期阻滞抑制K562细胞的增殖.

To explore the relationship between the expression levels of JARID1B,Hes1 and MMP-9 genes and the stages of chronic myelogenous leukemia(CML) and the curative effect of imatinib mesylate (IM).Peripheral blood samples of 15 cases of CML in chronic phase and 10 cases of CML in progressive phase were collected from the Hematology Department of Taihe Hospital affiliated to Hubei University of Medicine and 15 cases of healthy people in the Physical Examination Center. CML patients were divided into effective group and ineffective group based on the efficacy after treatment with IM, then real-time PCR was used to detect the expression levels of JARID1B, Hes1 and MMP-9 mRNA, finally, the differences in the level of gene expression and their correlations with CML stages and IM curative efficacy were analysed.The expression levels of Hes1 and MMP-9 in initially diagnosed patients in chronic and progressive phase without IM treatment were significantly higher than those of health people（P＜0.05）. There was no significant difference in the expression level of JARID1B between chronic phase patients and health people（P＞0.05）, but the expression level of JARID1B in the progressive phase patients was higher than that of health people （P＜0.05）. The expression levels of JARID1B and Hes1 in the IM-effective group were not significantly different from those in the IM-ineffective group (P=0.85,P=0.82）, while the expression level of MMP-9 in the IM-effective group [JP2]was significantly lower than that in the IM-ineffective group（P＜0.05）.The expression levels of JARID1B Hes1 and MMP-9 relate with the different phase of CML; The expression levels of JARID1B and Hes1 have not significant relationship with IM curative efficacy, the MMP-9 gene expression level relates with IM curative efficacy.伊马替尼治疗的慢性髓系白血病患者JARID1B、Hes1和MMP-9基因表达水平差异的研究.探讨JARID1B、Hes1和MMP-9基因表达水平与慢性髓系白血病（chronic myelogenous leukemia，CML）疾病的病程阶段及伊马替尼（imatinib mesylate, IM）疗效的关系.收集湖北医药学院附属太和医院血液科CML患者25例 (慢性期15例、进展期10例)和体检中心正常人15例的外周血标本。按初次确诊为CML患者的病期和服用IM后的疗效分组（有效组和无效组），采用实时定量RT-PCR方法检测JARID1B、Hes1和MMP-9基因mRNA表达水平，分析JARID1B、Hes1和MMP-9基因表达水平差异与疾病阶段及IM疗效的相关性。.初次确诊未服用IM慢性期和进展期CML患者Hes1和MMP-9基因的表达水平明显高于正常人（P＜0.05）；CML患者慢性期JARID1B基因的表达水平与正常人相比无明显差异（P=0.20），进展期患者JARID1B的表达水平明显高于正常人（P＜0.05）；IM治疗有效组JARID1B和Hes1基因的表达水平与无效组相比无明显差异（P=0.85、P=0.82），MMP-9的表达水平有效组明显低于无效组（P＜0.05）.JARID1B、Hes1和MMP-9基因的表达水平与CML的不同病程分期有关；JARID1B、Hes1基因表达水平与IM疗效无关，MMP-9基因表达水平与IM疗效有关.

To explore the values of 4 prognostic score systems in evaluation of clinical effecacy for patients with newly diagnosed chronic myeloid leukemia in chronic phase (CML-CP) treated with imatnib mesylate (IM) and the relationship between 4 prognostic score systems and deep molecular response (MR4.5).The clinical data of 240 CML-CP patients treated with imatinib mesylate in our hospital between Janunay 2008 and December 2017 were analyzed retrospecively. The risk was stratified according to 4 prognostic score systems, the relationship between the 4 prognostic score systems and 3-month early molecular response (3M-EMR), 6 month complete cytogenetic response (6M-CCyR), 12-month major molecular response (12M-MMR) as well as the correlation of the 4 prognostic score systems with deep molecalar response were analyzed.At the end of treatment for 3 months, the EMR was evaluated for 219 patients, among them 164 (74.9%) patients achieved 3M-EMR; at the end of treatment for 6 months, CCyR was evaluated for 180 pathsents, among them 130 (72.2%) patients achicved 6M-CCyR; at the end of treatment for 12 months, the MMR was evaluated for 111 patients, among them 60 (54.1%) patients achieved 12M-MMR. Compared with the high-risk group, the treatment response to IM in the low-risk group (including the low-risk group and the intermediate-risk group) was better. There was significant difference in 3M-EMR according to Sokal score and ELTS score (P＜0.05), and there was significant difference in 12M-MMR according to EUTOS score and ELTS score (P＜0.05). Logistic regression analysis revealed Sokal score (HR=0.69, 95%CI：0.22-1.37, P＜0.05) and 3M-EMR (HR=0.47, 95%CI：0.28-0.84, P＜0.01) independently related with MR4.5, The combination of Sokal score, especially the low risk with 3M-EMR much more can predict MR4.5 (HR=0.42, 95%CI=0.21-0.82, P＜0.01).There is a remarkable clinical efficacy of imatinib mesylate on CML-CP patients, moreover, low risk group has a better therapeutic response. Both Sokal score and ELTS score evaluate 3M-EMR better, both EUTOS score and ELTS score evaluate 12M-MMR better. The combination of low risk in Sokal score with 3M-EMR much more can predict MR4.5. The results of this study provide the reference basis for evaluating the clinical therapentic efficacy and timely modifying the therapeutic regimens for CML patients, also possess the reference value for predicting the MR4.5.不同评分系统对初诊慢性髓系白血病患者临床疗效的预估价值.探讨4种评分系统对甲磺酸伊马替尼（IM）治疗初诊慢性髓系白血病慢性期(CML-CP)患者临床疗效的预估价值，以及与深层分子学反应(MR4.5)之间的相互关系.回顾性分析我院2008年1月至2017年12月接受IM治疗的240例初诊CML-CP患者的临床资料，根据4种评分系统分别进行危险度分层，并分析4种评分系统与3个月早期分子学反应（3M-EMR）、6个月完全细胞遗传学反应（6M-CCyR）和12个月主要分子学反应（12M-MMR）之间的关系，以及4种评分系统与MR4.5之间的相关性.240例患者治疗满3个月时，有219例患者评估EMR，其中有164例（74.9%）患者获得3M-EMR；治疗满6个月时，有180例患者评估CCyR，其中有130例（72.2%）患者获得6M-CCyR；治疗满12个月时，有111例患者评估MMR，其中有60例（54.1%）患者获得12M-MMR。4种评分系统的低危组患者（低危+中危）相比高危组患者对IM的治疗反应性更好，其中Sokal及ELTS评分与3M-EMR差异有统计学意义（P＜0.05），EUTOS及ELTS评分与12M-MMR差异有统计学意义（P＜0.05）；多因素回归分析发现，Sokal评分（HR=0.69，95%CI：0.22-1.37，P=0.018）和3M-EMR（HR=0.47，95%CI：0.28-0.84，P＜0.01）与MR4.5独立相关，Sokal评分与3M-EMR结合能更好地预测MR4.5（HR=0.42，95%CI：0.21-0.82，P＜0.01）.IM治疗初诊CML-CP患者的临床疗效显著，低危组患者比高危组治疗反应更佳。Sokal及ELTS评分能较好地评估3M-EMR，而EUTOS及ELTS评分能较好地评估12M-MMR；Sokal评分的低危组与3M-EMR结合能更好地预测MR4.5。本研究结果为CML患者临床疗效的预估和及时更改治疗方案提供了参考依据，对预测MR4.5也具有参考价值.

To investigate the effect of stably down-regulating the FMI expression of K562 cells on the sensitivity of K562 cells to Imatinib (IM) and its possible mechanism.Western-blot was used to detect the expression of FMI protein in K562 cells and peripheral blood mononuclear cells from the patients with chronic myelogenous leukemia, chronic myeloid blast crisis and healthy volunteers. The specific interference sequences targeting at the human FMI gene were designed and ligated into the lentiviral vector LV3; the three plasmid system-packaged lentivirus particles were used to transfect K562 cells to screen K562 cells that stably down-regulated FMI. CCK-8 assay and flow cytometry were used to determine effect of IM on cell proliferation and apoptosis. The transcription level of FMI and Fz8 in leukemia cells was detected by fluorescent quantitative PCR. The protein expression levels of FMI, Fz8, NFAT1, BCR-ABL and β-catenin in leukemia cells were detected by Western-blot.The expression of FMI protein could be detected in peripheral blood mononuclear cells of the patients with CML-BC and K562 cells, the FMI expression could not be detected in all the patients with CML-CP and healthy volunteers. The recombinant lentiviral vector LV3/FMI had been successfully constructed the lentivirus was packaged, and the K562 cells stably down-regulating the FMI protein were screened. After stable down-regulation of FMI expression in K562 cells, the proliferation rate of leukemia cells decreased and the apoptosis rate was increased under the same drug concentration. Both the transcription and protein expression levels of Fz8 decreased. The NFAT1 total protein level increased, as well as the nuclear translocation of protein was enhanced. There was no significant change in the expression level of BCR-ABL fusion protein. The expression level of β-catenin protein decreased.After the stable down-regulation of FMI expression, the sensitivity of K562 cells to IM and apoptosis of cells increase, which are performed possibly by inhibiting the FMI-Fz8 signaling pathway and activating the Ca稳定下调K562细胞株FMI表达对其甲磺酸伊马替尼药物敏感性的影响.探讨稳定下调K562细胞株FMI表达对其甲磺酸伊马替尼药物敏感性的影响及其可能的作用机制.应用Western-blot方法分别检测K562细胞、慢性髓系白血病（CML）病人、慢性髓系急变病人及健康志愿者外周血单个核细胞FMI蛋白表达；设计靶向人FMI的特异性干扰序列，连接入慢病毒载体LV3；用三质粒系统包装慢病毒颗粒，感染K562细胞，筛选稳定下调FMI的K562细胞；应用CCK8法鉴定K562细胞对伊马替尼药物敏感性的变化，AnnexinⅤ/7-AAD双标记的流式细胞术检测细胞凋亡，荧光定量PCR检测白血病细胞FMI、Fz8的转录水平，Western-Blot方法检测白血病细胞FMI、Fz8、NFAT1、BCR-ABL、β-catenin的表达水平.在CML急变病人外周血单个核细胞与K562细胞可检测到FMI蛋白表达，在CML缓解期病人及健康志愿者外周血单个核细胞未检测到FMI表达。成功构建重组慢病毒载体LV3/FMI，并包装慢病毒，筛选出稳定下调FMI蛋白的K562细胞株；稳定下调K562细胞FMI表达后，在相同药物浓度作用下白血病细胞增殖率降低、凋亡率升高，Fz8基因转录水平降低，Fz8蛋白表达水平下降，NFAT1总蛋白水平增高，核转位增加，BCR-ABL融合蛋白表达水平无明显变化，总β-catenin蛋白表达水平下降.稳定下调FMI表达后，K562细胞对伊马替尼药物敏感性增高，凋亡增加，此作用可能是通过抑制FMI-Fz8信号通路而激活Ca

To investigate the clinical significance of SCIN gene expression and promoter methylation in patients with chronic myeloid leukemia (CML).Real-time quantitative PCR was used to detect the expression level of SCIN in mononucleatr cells of bone marrow samples from 64 CML patients and 37 controls. The methylation levels of SCIN promoter in 65 patients with CML and 29 controls were detected by real-time quantitative methylation-specific PCR and bisulfite sequencing PCR.The expression level of SCIN in CML patients was significantly down-regulated (P＜0.05), compared with the control group. The down-regulation rate of SCIN expression in CML patients at chronic phase, accelerated phase and blast crisis was 61%, 67% and 75%, respectively. Spearman correlation analysis showed that the expression level of SCIN negatively correlated with the transcript level of BCR-ABL1 (R=-0.315, P＜0.05). However, there was no significant difference in clinical parameters such as sex, age, white blood cell count, hemoglobin level, platelet count, chromosome, CML staging and BCL-ABL1 transcript level between low and high SCIN expression groups of CML patients (P＞0.05). No significant difference in methylation of SCIN promoter between CML patients and controls, and no correlation between SCIN expression and promoter methylation were observed (P＞0.05).The SCIN expression is down-regulated in CML patients, which may relate with the pathogenesis that is, BCR-ABL1 fusion gene induces CML tumorigenesis. The down-regulation of SCIN expression may relate with the progression of CML.慢性髓系白血病患者中基因SCIN表达及启动子甲基化的临床研究.探讨基因SCIN表达及启动子甲基化在慢性髓系白血病（CML）患者中的临床意义.应用实时定量PCR检测64例CML患者和37例对照骨髓单个核细胞SCIN的表达水平。应用实时定量甲基化特异性PCR结合亚硫酸盐测序PCR检测65例CML患者和29例对照SCIN启动子甲基化水平.CML患者与对照组相比细胞SCIN表达水平明显下调（P=0.010）；慢性期、加速期与急变期患者中SCIN表达下调率分别为61%、67%和75%，有增加趋势。Spearman相关性分析显示，基因SCIN表达与BCR-ABL1转录水平呈负相关（r=-0.315，P＜0.05）。然而，SCIN低表达组与高表达组相比较，在性别、年龄、外周血白细胞数、血红蛋白水平、血小板数量、染色体特征、CML分期及BCR-ABL1转录水平等临床参数中未见差异（P＞0.05）。CML患者SCIN启动子甲基化水平与对照组比较无统计学差异，且SCIN表达与启动子甲基化水平无相关性（P＞0.05）.基因SCIN在CML患者中非甲基化依赖性表达下调，可能与BCR-ABL1融合基因致病CML有关；SCIN表达下调可能与CML进展有关.

To investigate the pro-apoptotic effect and mechanism of miR-30a overexpression on chronic myeloid leukemia K562 cells.The k562 cells were transfected with the recombinant plasmid pEGFP-pre-miR-30a, the real-time quantitative PCR was used to detect the level of miR-30a and BCR/ABL, and then the cell apoptosis was assessed by flow cytometry with AnnexinV-FITC/PI double staining. Western blot was used to detect the expression of BCR/ABL protein,apoptosis-related protein BCL-2 and BAX, PTEN, AKT and p-AKT.Sequencing and digestion map indicated that the recombinant plasmid was constructed successfully. Compared with 2 control groups, the miR-30a expression in k562 cells transfected with recombinant plasmid pEGFP-pre-miR-30a was obviously up-regulated. The expression of BCR/ABL mRNA and BCR/ABL protein was both significantly down-regulated. Apoptotic rate was significantly enhanced (both P0.05),and the expression of anti-apoptotic protein BCL-2 was down-regulated while the expression of pro-apoptotic protein BAX was up-regulated. The level of PTEN was significantly up-regulated in omparison with control groups,no variation was found in total AKT, but the expression of p-AKT was down-regulated.The overexpression of miR-30a is abled to down-regulate the level of BCR/ABL mRNA and BCR/ABL protein, and increase apoptotic rate, its mechanism may be related with inhibition of the activity of BCR/ABL-PTEN/AKT signaling pathway.miR-30a的高表达促进K562细胞的凋亡.探讨miR-30a的高表达对慢性髓系白血病K562细胞株的促凋亡作用及其机制.构建pEGFP-pre-miR-30a重组质粒并转染K562细胞，采用实时定量PCR法检测miR-30a和 BCR/ABL mRNA的表达水平， 应用annexinV-FITC/PI双集流式细胞术（flow cytometry，FCM）分析K562细胞凋亡百分率，应用蛋白印迹法（Western blot）分析BCR/ABL融合蛋白、凋亡相关蛋白BCL-2、BAX及PTEN、AKT、p-AKT的变化.测序与酶切图谱证实成功构建重组质粒。与pEGFP-C1-K562阴性对照组和K562空白对照组比，重组质粒pEGFP-pre-miR-30a转染组miR-30a表达水平显著升高，BCR/ABL mRNA与蛋白表达明显下调，凋亡细胞所占比例明显增加（P0.05)，抗凋亡蛋白BCL-2水平降低，促凋亡蛋白BAX表达增加，抑癌蛋白PTEN表达明显上升，AKT无明显变化，但活性形式的p-AKT显著降低，差异均有统计学意义.高表达的 miR-30a能抑制K562细胞癌基因BCR/ABL mRNA及蛋白的表达并促进细胞凋亡，其机制可能与抑制 BCR/ABL-PTEN/AKT信号通路的活性有关.

As a potential target for cancer treatment, the C-Myc high expresses abnormally in a variety of solid tumors and hematological malignancies in humans. In hematologic malignancies, the increasing expression of C-Myc is associated with poor prognosis of patients diffuse large B-cell lymphoma. However, some studies have shown that high expression of C-Myc might be one of the mechanisms of disease progression and abrupt change, therefore, it can promote the transformation of chronic myeloid leukemia. Besides, the high expression of C-Myc may lead to the decrease of the sensitivity of chronic myelogenous leukemia cells to imatinib. Therefore, C-Myc has been concerned in the study of imatinib drug resistance. This review will focuses on the latest research progress on the mechanism of C-Myc in chronic myeloid leukemia in recent years.

To compare the immunomodulatory effects of the 2nd generation of tyrosine kinase inhibitors (TKIs)-dasatinib and nilotinib as well as the 1st generation of TKI-imatinib on chronic myeloid leukemia (CML) patients.To evaluate the T cell subtypes by flow cytometry on the CML patients of our center who received the treatment with dasatinib (n=10), nilotinib (n=26) or imatinib (n=44) for more than 3 months, and to analyze and correlate these data with the clinical remission situations and prognosis.80.0% of the patients in dasatinib group, 16.6% of the patients in nilotinib group and 27.5% of the patients in imatinib group respectively had a Th1 proportion in the peripheral blood (Th1/CD4This study shows that during the CML treatment with TKIs, the increased or normal Th1 proportion indicates a bigger chance for CCyR, MMR, and MR4.5. Dasatinib may significantly increase the level of Th1 while decrease the level of Treg in the patients, as compared with nilotinib and imatinib.

To explore the clinical significance of clonal evolution of additional chromosomal 8 in CML progression.An unusual case with the clonal evolution from trisomy 8 to tetrasomy 8 accompanied by 2 time of CML blast crisis (BC) was reported.This patient suffered from 2 time of CML blast crisis and the additional chromosome 8 aberrations were accompanied. Trisomy 8 and tetrasomy 8 were detected at first CML blast crisis and second CML blast crisis, respectively. After tetrasomy 8 was developed, the c-Myc was over-expressed and the central nervous system leukemia happened in this case. Only high dose Ara-C and MTX regimen could induce remission for a short period.These findings suggested that additional chromosome 8 aberrations are important marker for poor prognosis of CML patients and contribute to a poor prognosis.

To analyze the relation between the signle nucleotide polymorphisms (SNP) of CYP3A5 gene and MDR1 gene loci and the risk of cytogenetic relapse in chronic myeloid leukemia (CML).The clinical data of 90 patients with CML treated with imatinib in our hospital were collected.The patients were divided into 2 groups: non-relapse and relapse according to relapse and non-relapse, then the relation between the SNP of CYP3A5 gene and MRD1 gene loci and the risk of cytogenetic relapse in CML patients.The grouping result showed that the patients with non cytogenetic relapse accounted for 41 cases those were enrolled in non-relapse group, and patient-with cytogenetic relapse accounted for 49 cases those were enrolled in relapse group. The follow-up time was 36 months. The detection showed that the incidence of cytogenetic relapse in the patients with CC genotype was significantly higher than that in the patients with TT+CT genotype of C3435T and C1236T at MDR1 gene loci (P0.05).Compared with the patients with CT+CC genotype in C3435T locus of MDR1 gene, the rate of cytogenetic relapse in the patients with TT genotype decreased significantly (P0.05). Compared with patients with CT+CC phemotype of C3435T in MDR1 gene locus, the non-relapse survival time of TT genotypes was significantly prolonged (P0.05). Compared with non-relapse group, the incidence of neutropenia (29.27% vs 71.43%) and blood toxicity (39.02% vs 61.22%) in the relapse group increased significantly (P0.05). The imatinib dose (OR=2 95, 95% CI:1.37~7.76) and the C3435T genotype in MDR1 genes (OR=0.09, 95% CI:0.05~0.72) were the factors affecting the cytogenetic relapse of the patients with CML (both P0.05).The therapeutic dose of imatinib and the C3435T and C1236T genotypes in MDR1 gene have a certain effect on the cytogenetic relapse of CML patients. C3435T genotypes in the.MDR1 gene showed a certain predictive value for evaluating the risk of cytogenetic relapse, which can be used as a clinical biomarker.

To explore the effect of additional chromosomal abnormalities on the prognosis and outcome of CML-CP patients receiving imatinib therapy.The clinical and genetic data of 589 CML-CP patients receiving imatinib treatment between May 2009 and October 2014 in the 1st Affiliated Hospital of Soochow University were analyzed, the 589 patients were divided into 5 groups according to the karyotypes at the initial diagnosis. The OS(overall survival), PFS (progression-free survival), EFS (event-free survival), Cumulative MMR (major molecular remission) and Cumulative CCyR (complete cytogenetic remission) were calculated by using the Kaplan-Meier method and compared by using the log-rank text by Graphpad 6.0. The χThere was significant difference about the frequency of optimal molecular response at 3 and 6 months between CML-CP patients with additional chromosomal abnormalities and those with classic t(9;22) ［50%（12/24） vs. 73.94%(261 /353), P0.05; 50%(10 /20) vs. 72.05%(232 /322) (P0.05)］, and the same significant difference was found at 6 months between the group with variant translocations and that with classic t(9;22) ［53.3% (16 /30) vs. 72.05%(232 /322) (P0.05)］. The P values of cumulative CCyR (P0.05) and EFS (P0.01) for 4 years were statistically significant between CML-CP patients with additional chromosomal abnormalities and the other 4 groups. Compared one to another, there was the significant difference in cumulative CCyR and EFS for 4 years between CML-CP patients with additional chromosomal.abnormalities and those with classic t(9;22) (47.25% vs. 84.01%)(P0.05); (75.03% vs. 90.01%)(P0.01).The additional chromosomal abnormalities influence the outcome of CML-CP patients receiving imatinib treatment, which make poor prognosis.

To evaluate the long-term prognosis of CML patients whose BCR-ABL transcript level was warning and best response at 12 months of treatment with tyrosine kinase inhititor (TKI), and to investigate the factors affecting therapeutic efficacy and prognosis.The clinical data of patients with newly diagnosed CML were analyzed retrospectively. According to BCR-ABL transcript level, the 80 patients were divided into group A and group B, the patients with BCR-ABLThe MMR and MRPatients at 12 months warning effect are slower and less easier to get DMR, which has a poor long-term prognosis. The size of the spleen in the patient at warning for 12 months of treatment effect can predict the relatively poor long-term prognosis. For a patient with a 12 months response to the warning, an early replacement therapy is available on the basis of combining other factors..

To explore the individualized treatment for patient with chronic phase chronic myeloid leukemia(CML-CP).The clinical data and treatment process of one CML-CP patient which intolerated to nilotinib were analyzed.Nilotinib was given to the patient once the diagnosis of CML-CP was set. Although major molecular remission (MMR) and complete cytogenetic remission (CCyR) were obtained during treatment for 3 months, a grade 3-4 hepatotoxicity appeared in the course of treatment.With drug reduction and symptomatic treatment, nilotinib was discontinued after 3 withdrawals and replaced with imatinib in January 11, 2015. The patients achieved MMR and CCyR at 7 months after imatinib replacement. At present, the patient tolerated well without any adverse events.Imatinib can be used as a second-line treatment drug for CML patients who was intolerant to nilotinib, and with less adverts, good effect and so on.

The past studies found that the treatment of chronic myeloid leukemia (CML) with imatinib can induce the macrocytic anemia, moreover the incidence of anemia increases along with enhancement of imatinib concentration. This study was aimed to evaluate the potential relation of erythrocyte mean corpuscular volume (MCV) increase after the treatment with tyrosine kinase inhibitors (TKI) with the therapeutic response in patients with CML-chronic phase (CML-CP).The clinical and hematologic data including MCV, molecular and cytogenetic response of 119 patients with CML-CP were collected after treatment with TKIs, and the relation of MCV changes after treatment with the clinical characteristics and therapeutic efficacy for patients with CML-CP was analyzed.The MCV in patients treated with TKIs for 12 months significantly increased as compared with that at initial diagnosis (P0.05). The proportion of patients with increased MCV in group of complete cytogenetic response (CCyR) was significantly higher than that in group of non-CCyR (P0.05). As compared with decreased MCV group, the patients in increased MCV group much more easily achieved CCyR after treatment for 6, 12 months (P0.05, P0.05) respectively, furthermore, much more easily maintained MMR (P0.05).The MCV as a parameter which is easily acquired may be a new marker for prodecting the therapeutic response of patients treated with TKIs.

β-catenin is an important member of Wnt signaling pathway. With more and more research on the relationship between β-catenin and cancers, it is noted that the concentration of β-catenin in cells is not only the landmark of activating the Wnt signaling pathway, but also a common phenomenon in various tumor cells. Moreover, abnormal expression of β-catenin is reported in blastic phase of CML and IM-resistant CML. Besides, β-catenin inhibitor is proved to be effective for the improvement of the IM susceptibility and the postponment of blastic phase. Here is to make a brief review about the mechanisms of β-catenin in the treatment of CML, such as classical signaling pathways as well as new genes and pathways.