目的:观察F框/WD40域蛋白7(Fbxw7)在急性T淋巴细胞白血病中的表达,探讨其对细胞生长的影响及其机制.方法:采用逆转录聚合酶链反应(RT-PCR)检测62例急性T淋巴细胞白血病患者中的Fbxw7 mRNA表达情况,将携带Fbxw7的过表达慢病毒载体转至白血病Jurkat细胞后,采用RT-PCR和Western blot检测其转染效果,MTT法检测Jurkat细胞增殖情况,流式细胞仪检测细胞周期变化和细胞凋亡情况,Western blot检测c-Myc蛋白和周期蛋白E(Cyclin E)的表达.结果:急性T淋巴细胞白血病复发组患者和初发组患者中Fbxw7 mRNA表达水平较正常健康人明显降低(P＜0.05);病毒转染后,Fbxw7组(转染携带Fbxw7过表达慢病毒载体)中Fbxw7 mRNA和蛋白质的相对表达水平较对照组(未转染的细胞)、阴性组(转染对照慢病毒载体)明显升高(P＜0.05);转染24 h后,细胞增殖受影响不明显(P＞0.05),而转染48 h和72 h后,细胞增殖明显受到抑制(P＜0.05);转染48 h后,与对照组相比,Fbxw7组Jurka细胞中G0/G1期细胞所占比例和细胞凋亡率明显升高,S期和G2/M期细胞所占百分比、c-Myc和Cyclin E蛋白的相对表达量明显降低,差异均有统计学意义(P＜0.05).结论:Fbxw7在急性T淋巴细胞白血病中低表达,上调其表达可抑制白血病Jurkat细胞增殖,促进细胞凋亡,其作用机制可能与下调c-Myc和Cyclin E蛋白表达有关.

目的 探究人类白细胞抗原-DR(HLA-DR)和钙结合蛋白(S100A12)在血液肿瘤合并脓毒血症患者中的诊断和预后价值.方法 回顾性分析60例血液肿瘤患者的临床资料,根据是否并发脓毒血症分组,A组18例患者并发脓毒血症,其余42例患者未并发脓毒血症,设为B组,分析血液肿瘤并发脓毒血症危险因素.采用流式细胞仪对2组患者血清中的HLA-DR、S100A12进行检测,分析检测影响.结果 A组死亡率(44.44％)、急性生理学及慢性健康状况(APACHEⅡ)评分、S100A12平均水平高于B组,A组HLA-DR水平低于B组(P＜0.05);CRP、S100A12、APACHEⅡ、HLA-DR均与血液肿瘤并脓毒血症患者的预后存在相关性(P＜0.05),HLA-DR、S100A12高表达是提示血液肿瘤并发脓毒血症预后死亡的独立指标.结论 HLA-DR和S100A12是预后死亡的重要指标,对血液肿瘤合并脓毒血症患者预后判断具有重要诊断价值,值得临床推广和运用.

目的 探讨骨髓中缺氧诱导因子1 α、血小板衍生生长因子B在骨髓增生异常综合征的表达.方法 收集我院46名骨髓增生异常综合征患者作为观察组,以我院健康人群46例作为对照组.对比观察组和对照组骨髓中缺氧诱导因子1α和血小板衍生生长因子B的表达水平.结果 观察组骨髓中缺氧诱导因子1 α和血小板衍生生长因子B表达水平明显高于对照组(P＜0.05);5q-综合征、难治性血细胞减少伴单系发育异常、难治性贫血伴原始细胞增多-1 和难治性贫血伴原始细胞增多﹣2 患者中缺氧诱导因子1 α及血小板衍生生长因子B表达水平存在差异,以难治性贫血伴原始细胞增多﹣2表达最高(P＜0.05 );预后死亡者缺氧诱导因子1 α、血小板衍生生长因子B表达水平高于存活患者,差异有统计学意义(P＜0.05 ).结论 通过测定骨髓增生异常综合征患者骨髓中缺氧诱导因子1 α、血小板衍生生长因子B的表达水平,可以对疾病的分型及预后进行较好的判断.

目的 探讨血清糖类抗原153(CA153)检测在巨幼细胞贫血(MA)病程中的变化.方法 选取入院前未接受叶酸(FA)、维生素B12(VitB12)补充治疗的MA患者48例,作为研究组.选取体检健康者32名,作为对照组.比较研究组治疗前后与对照组的CA153水平,并分析CA153与其他检测指标的相关性.结果 研究组治疗前CA153水平为(51.10±5.77)U/mL,对照组CA153水平为(20.22±4.69)U/mL(P<0.001).经2～6周FA、VitB12补充治疗后,CA153水平显著下降[(21.92±4.52)U/mL](P<0.001),与对照组比较差异无统计学意义(P=0.108).CA153与血红蛋白(Hb)之间呈负相关(r=-0.882,P<0.001),CA153与乳酸脱氢酶(LDH)之间呈正相关(r=0.455,P=0.001).结论 MA患者血清CA153水平升高,血清CA153检测可为MA的早期诊断提供参考依据.

目的 探究艾拉莫德联合甲氨蝶呤治疗活动性类风湿关节炎的疗效及安全性.方法 选择我院2016年1月～2018年1月期间治疗的80例活动性类风湿关节炎患者为研究对象,回顾性分析其临床记录资料.按照随机原则将80例患者等分为观察组和对照组,各40例,对照组每次口服甲氨蝶呤15mg,每周1次,观察组在对照组基础之上联合口服艾拉莫德25mg,饭后服用,早晚2次,对两组患者的治疗效果进行对比分析,观察比较其安全性和不良反应.结果 治疗三个疗程后,观察组ACR20、ACR50、ACR70有效率分别达到85.00％、57.50％和52.50％,对照组ACR20、ACR50、ACR70有效率分别为50.00％、37.50％和25.00％,两组之间数据比较差异较大,表现出统计学意义(P<0.05).观察组肝功能异常、皮疹等不良反应发生率为12.50％明显低于对照组不良反应发生率37.50％,两组之间数据比较差异明显,存在统计学意义(P<0.05).结论 艾拉莫德联合甲氨蝶呤治疗活动性类风湿关节炎临床效果显著,其疗效由于单纯甲氨蝶呤药物治疗,安全性较好,降低了不良反应发生率,值得在临床治疗中推广应用.

Objective:To investigate the effect of miR-21 on the proliferation and apoptosis of leukemia cell line K562 and the regulation of PI3K/AKT signaling pathway.Methods:K562 cells were divided into control group,miR-21 NC group and miR-21 interference group,the first group without treatment,the two groups after using cationic liposome transfection of LipofectamineTM2000 miR-21 inhibitor and miR-21 negative control.After transfection 48 h,the expression of miR-21 mRNA in cells was detected by Real-time fluorescence quantitative (qRT-PCR),MTT assay was used to test the effect of miR-21 on cell proliferation,flow cytometry was used to detected the effect of miR-21 on cell cycle apoptosis rate,the protein expression of PI3K,AKT and p-AKT in PI3K/AKT signaling pathway after miR-21 transfection were detected by protein immunoblotting (Western blot).Results:The expression level of miR-21 mRNA and the survival rate of cells in the miR-21 interference group were significantly lower than those in the control group and the miR-21 NC group;compared with control group and miR-21 group NC,flow cytometry detected the cells accounted for proportion of G0/G1 phase in miR-21 interference group increased significantly,the proportion of cells at S phase significantly decreased,and apoptosis rate increased significantly.Western blot test results show that the expression levels of p-AKT in miR-21 inter-ference group were significantly lower than those in the control group and miR-21 NC group,but the expression levels of PI3K and AKT protein were not changed significantly.Conclusion:Down-regulation of miR-21 can inhibit the proliferation and promote apoptosis of leukemia K562 cells,and its mechanism may be related to the inhibition of PI3K/AKT signaling pathway.

近十余年来慢性粒细胞白血病(CML)的分子生物学研究不断有新的进展,其中之一就是陆续报道了二十余例BCR (breakpoint cluster region)基因上的断裂点不在M-bcr (major breakpoint cluster region)而位于m-bcr (minor breakpoint cluster region)、表达相对分子质量为190 000 BCR-ABL融合蛋白的CML (p190 CML).p190 CML是一个罕见的CML分子亚型,以下就p190 CML的发现、发病率、临床和血液学特征、治疗和临床转归及临床和生物学意义等方面的研究作一综述.

树突状细胞(DC)是免疫调节的关键细胞,为唯一能提呈抗原给初始型T细胞并诱导其活化和功能分化的细胞,可作为治疗恶性肿瘤或感染等疾病的细胞疫苗.随着基于DC的免疫治疗进入临床研究,稳定地产生足量有效的DC是技术关键.获得DC的方法除直接分离纯化外,还可用细胞因子(CK)体外培养.前一方法产量很低,后一方法也有其不尽如人意之处,如培养时间较长以及有的急性髓系白血病(AML)细胞不能被CK诱导分化为DC.虽然有报道乙酸豆蔻佛波酯(PMA)也能诱导CD34+细胞分化为DC,但由于PMA的促癌作用,不能用于临床.因此有必要寻找其他获得DC的方法.最近发现钙动员(calcium mobilization)可诱导髓系细胞变成DC,以下就钙动员和CK诱导正常和恶性髓系细胞分化为DC的比较研究以及钙动员诱导DC分化的信号转导机制的研究等方面的内容作一综述.

OBJECTIVES:To explore the feasibility of DC being in vitro induced from AML cells with cytokine cocktails and their biological properties.METHODS:AML cells were cultured in either presence or absence of cytokine cocktails. DC were studied for morphology, and cytochemical and immunofluorescent staining. Functions of DC were examined by MLC, FITC-conjugated dextran uptake test, and LDH release assay. RT-PCR and FISH were used to analyze the specific fusion genes of culture-derived DC.RESULTS:Classical DC morphological changes occurred in all 15 cultured AML cells. DC-associated surface molecules such as CD(1a), CD(80), CD(86), CD(106), CD(83) and HLA-DR were upregulated (P < 0.05). The allostimulatory abilities of culture-derived DC were significantly higher than those of AML cells uncultured or cultured in the absence of cytokines (P < 0.05). Culture-derived DC only in the presence of GM-CSF + IL-4 have phagocytotic activities. CTL assay was performed in 5 of the 15 samples. At effector/target ratio of 20:1, auto-T lymphocytes primed with the culture-derived DC exhibited no more killing activity to auto-AML cells than those stimulated by IL-2 or uncultured AML cells. Culture-derived DC presenced the native AML-specific aberrant karyotype and related fusion gene.CONCLUSIONS:Cytokine cocktails could in vitro induce AML cells into DC with classical morphology, immunophenotype and function. DC maturity induced by different cytokine cocktails could be variable. Culture-derived DC were originated from the native AML cells. AML cells could make the auto-T lymphocyte anergy.

微量残留病(MRD)是导致急性髓系白血病(AML)复发的根本原因，人们一直在不断找寻根除MRD的方法。大量研究表明细胞免疫治疗方法已经成熟，可根除MRD从而治愈AML。AML细胞在体内不能有效被T细胞识别，主要机制为70％的AML细胞表面缺乏共刺激分子CD(80)和CD(86)(分别为T细胞共刺激受体CD(28)和CTLA-4的配体)，不能提供T细胞活化必需的第二信号，从而导致肿瘤特异性T细胞克隆……