急性粒细胞白血病基因表达谱的研究

目的在急性粒细胞白血病M2a亚型的治疗中,首次完全缓解持续时间(CCR)是和预后相关的最重要的指标。运用基因芯片技术,研究拥有不同CCR病人的基因表达谱差异,从而寻找影响AML预后相关基因。方法收集3例CCR6个月内复发(A组)和3例CCR12个月以上(B组)的AML患者初诊时骨髓单个细胞,纯化mRNA,分别用Cy3和Cy5标记,和Agilent Human1B寡核苷酸基因芯片杂交,研究基因表达谱差异。结果在检测的20173个基因中,筛选出共同差异表达基因21个,其中在A组各例都表达上调的基因有10个,同时表达下调的基因有11个。结论 APP等21个基因与采用标准化疗的AML首次完全缓解持续时间有关,这些基因可能成为早期诊断难治性AML和判断预后的指标。     

三氧化二砷诱导NB4细胞凋亡相关基因表达改变的基因芯片研究

目的 利用基因芯片技术研究NB4细胞经三氧化二砷(As2O3)诱导凋亡后的凋亡相关基因表达变化.方法 检索NCBI的Entrez以及Human IPI数据库,通过染色体定位来排除冗余基因,共获得1 384个凋亡相关基因.探针使用软件OligoArray 2.0设计,并作Blast比对.设计基因特异寡核苷酸探针,合成后点样,制备寡核苷酸芯片.用2μmol/L的As2O3处理NB4细胞48h后,提取细胞总RNA,分别以Cy3,Cy5标记对照组及实验组,随后与含1 384个凋亡相关基因的寡核苷酸芯片杂交,使用基因芯片扫描仪对杂交信号扫描,软件分析Cy3和Cy5两种荧光信号的强度和比值,找出经As2O3处理后出现差异表达的基因,并挑选其中差异表达最明显的4条基因,设计引物后与CNDA产物进行PCR扩增,琼脂糖凝胶电泳进行验证.结果 NB4细胞经2μmol/L As2O3作用48h后共有4个基因表达上调,12个基因表达下调,且RT-PCR验证结果与基因芯片结果完全相符.结论 As2O3可以诱导NB4细胞一系列基因表达的改变,这些涉及信号转导、转录调节、细胞周期、氧化应激、蛋白质的翻译合成及细胞分化等方面基因的差异表达,可能在NB4细胞凋亡中起着重要作用.     

耐药性颞叶癫痫脑组织中隆纹菌素的表达及临床意义

目的 研究耐药性颞叶癫痫患者脑组织中隆纹菌素(STRN)的表达,探讨其在耐药性颞叶癫痫发病中的意义.方法 从重庆医科大学附属第一医院神经内科建立的耐药性癫痫患者脑组织库中随机抽取42例耐药性颞叶癫痫患者术后脑组织.采用基因芯片检测STRN eDNA的表达,以Cy3-dCTP标记对照组cDNA,以Cy5-dCTP标记实验组eDNA,收集荧光信号,Cy5/Cy3*(Cy3*为校正后的Cy3信号值)小于0.5或大于2.0提示基因mRNA有表达差异.采用免疫组织化学、免疫荧光、Western blotting检测STRN的蛋白表达,并与对照组(12例,均为行手术减压或清创患者的颞叶脑组织)进行比较.结果 耐药性颞叶癫痫患者颞叶脑组织中编码STRN的基因出现低表达(Cy5/Cy3*值为0.384).STRN蛋白在耐药性颞叶癫痫患者颞叶脑组织中的表达量与对照组相同部位比较明显降低:免疫组化显示实验组光密度值(0.310 8±0.108 7)明显低于对照组(0.421 04±0.120 2,P<0.05);免疫荧光检测亦显示实验组光密度值(0.026 04±0.023 2)明显低于对照组(0.050 6±0.001 1,P<0.05);Western blotting检测显示实验组STRN蛋白表达(光密度值0.591 4±0.219 7)明显低于对照组(0.980 4±0.140 1,P<0.05).结论 耐药性颞叶癫痫患者颞叶脑组织中STRN表达明显降低.STRN表达降低及其引发的雌激素信号转导、内源性一氧化氮合成减弱在耐药性颢叶癫痫发病中可能有重要作用.     

淋巴瘤患者外周血单个核细胞FOXp3与HLA-B基因表达研究

 目的 探讨淋巴瘤患者外周血单个核细胞(peripheral blood mononuclear cells, PBMC)FOXp3与人白细胞抗原B (human leukocyte antigen, HLA-B),的基因表达水平及对淋巴瘤的临床应用价值.方法 设计FOXp3和HLA-B基因引物和实时荧光PCR相对定量法反应体系,在实时荧光定量PCR仪上检测69例淋巴瘤患者和30例健康人外周血单个核细胞内基因表达水平并分析其差异.结果 淋巴瘤患者FOXp3和HLA-B的mRNA相对表达量分别为1.46±0.45和0.74±0.22,健康组分别为1.15±0.30和1.05±0.28,两组差异均有统计学意义.在淋巴瘤患者组中,Ⅰ期+Ⅱ期的HLA-B相对表达量高于Ⅲ期+Ⅳ期,差异有统计学意义,而FOXp3未见区别.结论 淋巴瘤患者外周血单个核细胞的FOXp3基因表达水平上调,HLA-B的基因表达水平下调且HLA-B不同分期间存在差异. 检测PBMC的FOXp3、HLA-B基因表达水平有助于淋巴瘤诊断和研究.     

NF-kB抑制与X射线诱导的人非霍奇金淋巴瘤细胞凋亡的研究

目的 探讨NF-kB p65对X射线诱导人非霍奇金淋巴瘤(non-Hodgkin lymphoma,NHL)细胞凋亡的作用及其调控机制.方法 以NF-kB p65抑制剂quinazoline (QNZ)处理人NHL细胞.将NHL细胞株Namalwa、Ramos和Raji细胞分为空白对照组、单纯照射组(IR)和X射线+QNZ实验组( IR+ QNZ),采用Annexin-V染色方法检测肿瘤细胞凋亡水平的变化,采用Western blot方法检测各细胞株中Survivin及凋亡相关蛋白Bax、Bcl-2和Cleaved Caspase-3的表达水平;应用实时定量PCR方法检测Survivin mRNA水平.结果 应用流式细胞仪检测凋亡细胞百分比,结果显示,QNZ处理后进行照射与单纯照射组相比凋亡细胞明显增加,且具有药物浓度依赖性(t=2.93～12.52,P＜0.05),Western blot法检测结果显示,电离辐射可显著增加人NHL细胞中Survivin蛋白的表达.而应用1、10和50 nmol/L QNZ处理人NHL细胞24 h后进行照射,可显著下调电离辐射所诱导的NHL细胞中的Survivin蛋白表达.随药物浓度增加其作用更为明显(t=3.29～ 16.72,P＜0.05).同时凋亡相关蛋白Bcl-2表达减低,而Bax和Cleaved caspase-3蛋白表达增加,Bcl-2/Bax比值明显降低,与单纯照射组相比,差异有统计学意义(t =6.20 ～9.91,P＜0.05).预处理QNZ后射线诱导的3种NHL细胞Survivin mRNA均不同程度下降.结论 抑制NF-kB能够增加X射线诱导的NHL细胞凋亡,其机制可能与下调Survivin蛋白表达水平及对凋亡相关蛋白Bcl-2家族的调节有关.     

应用基因芯片筛查妊娠期高血压疾病相关基因初探

 目的 比较人正常妊娠胎盘组织及妊娠期高血压疾病胎盘组织基因表达的差异性,寻找PIH相关基因,以进一步探讨妊娠期高血压疾病的发病机制.方法 从正常妊娠胎盘和PIH胎盘中抽提mRNA,反转录后,分别用Cy3、Cy5荧光标记,获得两组胎盘来源的cDNA探针.cDNA探针与含8 190条人cDNA点样的BioDoor基因芯片杂交,结果由激光共聚焦扫描仪扫描,并用软件进行统计分析.结果 与正常妊娠胎盘表达的基因相比,PIH胎盘组织中有393条(4.79%)的基因表达发生了变化,其中197条基因在PIH时表达量降低,而196条基因在妊娠期高血压疾病时表达量增高.结论 基因芯片技术是研究PIH发病机制的一种新方法.     

18 F-FDG PET显像对不同亚型淋巴瘤的诊断价值

目的：探讨^18F-脱氧葡萄糖(FDG)PET显像对霍奇金淋巴瘤(HL)和以世界卫生组织(WHO)分类标准分类的不同亚型非霍奇金淋巴瘤(NHL)的诊断价值。方法：对236例淋巴瘤(62例HL和174例NHL)患者的FDG PET全身显像结果进行回顾性分析，并与WHO病理分型的结果比较。结果：PET显像对淋巴瘤的检出阳性率为94％(221／236例)，对HL和NHL的阳性率分别为97％(60／62例)和93％(161／174例)。在不同NHL亚型中，8例套细胞淋巴瘤，99％(76，77例)的弥漫性大B细胞淋巴瘤(DLBCL)，95％(55／58例)的滤泡性淋巴瘤(FL)，73％(8／11例)的淋巴结边缘区淋巴瘤(MZL)，2／3例黏膜相关性(MALL型)结外边缘区B细胞淋巴瘤(MALT-MZL)，5／8例的无其他特征外周T细胞淋巴瘤(PTCL)，2／3例的伯基特淋巴瘤(BL)，2例间变性大细胞性淋巴瘤和覃样肉芽肿、小淋巴细胞性淋巴瘤和NK／T细胞型淋巴瘤各1例FDG摄取异常，而13例(3例MZL，3例PTCL，3例FL，MALT-MZL、DLBCL、BL和前体T淋巴母细胞淋巴瘤各1例)未见异常FDG分布。结论：^18F-FDG PET显像对常见的NHL亚型检出阳性率较高，对相对少见的NHL亚型检出阳性率较低。     

LON基因下调对人乳腺癌细胞系MCF-7的增殖以及药物敏感性的影响

目的：应用RNAi的方法抑制丝氨酸蛋白酶LON表达，研究该丝氨酸蛋白酶的表达与肿瘤增殖以及药物敏感性之间的关系。方法：以人乳腺癌细胞MCF-7为研究对象，通过RNAi下调LON基因表达，采用RT—PCR法、MTT增殖实验、流式细胞术等技术进行检测。结果：在瞬时转染与稳定转染细胞株中，RT—PCR结果证实LON基因的表达明显下调；MTT增殖实验结果显示，LON基因的下调可导致细胞的增殖能力降低，并且能够增加对化疗药顺铂的药物敏感性。流式细胞仪检测显示LON基因的下调对细胞周期没有显著影响。结论：通过RNAi下调LON基因能够抑制肿瘤细胞的增殖并且与顺铂有协同作用，其抑制作用未造成细胞周期的改变。     

口咽部非霍奇金淋巴瘤的CT诊断

目的 分析口咽部非霍奇金淋巴瘤(non-Hodgkin's lymphoma,NHL)的CT表现特征,探讨各种不同病理类型NHL的CT表现特点. 资料与方法 回顾性分析33例经病理证实的口咽部NHL患者的CT表现. 结果 33例中,B细胞来源18例(54.5%),T和NK细胞来源15例(45.5%).病变分布:扁桃体14例(弥漫性大B细胞淋巴瘤8例、NK/T细胞淋巴瘤5例、套细胞淋巴瘤1例);舌根10例(弥漫大B细胞淋巴瘤5例、滤泡性淋巴瘤3例、血管免疫母细胞性T细胞淋巴瘤1例、周围T细胞淋巴瘤1例);软腭2例(NK/T细胞淋巴瘤1例,结外边缘区B细胞淋巴瘤1例);累及两个以上口咽部位7例,均为NK/T细胞淋巴瘤.33例中,肿块型18例,其中13例为弥漫大B细胞淋巴瘤;弥漫型10例,其中8例为NK/T细胞淋巴瘤;无明显异常CT表现者5例. 结论 口咽NHL多发生于腭扁桃体及舌根,病理类型以弥漫大B细胞淋巴瘤和NK/T细胞淋巴瘤为主;CT对于NHL的鉴别诊断和病变范围的判断有重要价值.     

人LDLR基因的克隆及在Hepa1-6细胞中的表达

目的 :从能感染HCV的人肝癌细胞HepG2中克隆出人LDLR基因 ,构建其真核表达质粒 ,并在小鼠肝癌细胞Hepa1 6中进行表达。方法 :从HepG2中提取总RNA ,采取逆转录PCR(RT PCR)方法得到人LDLR的cDNA。将获得的人LDLR基因克隆到真核表达载体pcDNA3中 ,进行酶切和序列鉴定。将重组质粒pcDNA3 hLDLR和空载体转入Hepa1 6 ,G4 18筛选 ,并用RT PCR和FACS检测人LDLR基因转录和蛋白的表达。结果 :人LDLR的cDNA已插入载体pcDNA3构建成真核表达质粒pcDNA3 hLDLR ,双酶切和测序表明 ,克隆的人LDLR与已登录GenBank的序列存在核苷酸多态性 ,但编码的氨基酸序列完全一致 ,该序列的GenBank登录号为gi|2 16 2 96 4 7|gb|AY114 15 5 .1,并且真核表达质粒的构建正确。用脂质体介导的方法转入Hepa1 6后 ,用RT PCR和FACS检测表明 ,细胞可表达人LDLR。结论 :成功地构建了真核表达质粒pcDNA3 hLDLR ,为进一步研究HCV和人LDLR的相互作用 ,以及建立可能的HCV细胞感染模型奠定了基础。     

Knee injury and Osteoarthritis Outcome Score (KOOS) - validation of a Swedish version

The Knee injury and Osteoarthritis Outcome Score (KOOS) is a self-administered instrument measuring outcome after knee injury at impairment, disability, and handicap level in five subscales. Reliability, validity, and responsiveness of a Swedish version was assessed in 142 patients who underwent arthroscopy because of injury to the menisci, anterior cruciate ligament, or cartilage of the knee. The clinimetric properties were found to be good and comparable to the American version of the KOOS. Comparison to the Short Form-36 and the Lysholm knee scoring scale revealed expected correlations and construct validity. Item by item, symptoms and functional limitations were compared between diagnostic groups. High responsiveness was found three months after arthroscopic partial meniscectomy for all subscales but Activities of Daily Living.     

Endovascular management of carotid-cavernous fistulas

Objective To investigate endovascular treatment of traumatic direct carotid-cavernous fistulas （CCF） and their complications such as pseudoaneurysms. Methods： Over a five-year period, 22 patients with traumatic direct CCFs were treated endovascularly in our institution. Thirteen patients were treated once with the result of CCF occluded, 8 twice and 1 three times. Treatment modalities included balloon occlusion of the CCF, sacrifice of the ipsilateral internal carotid artery with detachable balloon, coll embolization of the cavernous sinus and secondary pseudoaneurysms, and covered-stem management of the pseudoaneurysms. Results All the direct CCFs were successfully managed endovascularly. Four patients developed a pseudoaneurysm after the occlusion of the CCF with an incidence of pseudoaneurysm formation of 18.2% （4/22）. A total number of 8 patients experienced permanent occlusion of the ICA with a rate of ICA occlusion reaching 36.4% （8/22）. Followed up through telephone consultation from 6 months to 5 years, all did well with no recurrence of CCF symptoms and signs. Conclusion Traumatic direct CCFs can be successfully managed with endovascular means. The pseudoaneurysms secondary to the occlusion of the CCFs can be occluded with stent-assisted coiling and implantation of covered stents.     

The acutely limping child

Acute limping may be the result of multiple pathologies in children. The differential diagnosis varies based on the age of the child. Irrespective of age, the initial imaging work-up includes AP and frog leg radiographs of the pelvis and ultrasound; MRI may sometimes be helpful. In children less than 3 years, infections and trauma are most frequent. MRI is the imaging modality of choice when osteomyelitis is clinically suspected. Between the ages of 3 and 10 years, transient synovitis of the hip and Legg-Calvé-Perthes disease are main considerations but infection, inflammation and focal bony lesions are also considered. In children over 10 years, slipped capital femoral epiphysis also is considered.     

Do Balance Board Training Programs Reduce the Risk of Ankle Sprains in Athletes?

Introduction Ankle sprains are the most common musculo-skeletal injury that occurs in athletes,particularly in sports that require jumping and landing on one foot such as soccer,and basketball(1-4).These injuries often result in significant time loss from participation,long-term disability,and have a major impact on health care costs and resources(5-8).     

High Intensity Interval Training:New Insights

KEY POINTS ·High-intensity interval training(HIT)is characterized by repeated sessions of relatively brief，intermittent exercise.often performed with an“a11 out”effort or at an intensity close to that which elicits peak oxygen uptake(i.e.，≥90%of VO2 peak).     

Developmental validation of the PowerPlex(®) ESI 16 and PowerPlex(®) ESI 17 Systems: STR multiplexes for the new European standard

In response to the ENFSI and EDNAP groups’ call for new STR multiplexes for Europe, Promega^® developed a suite of four new DNA profiling kits. This paper describes the developmental validation study performed on the PowerPlex^® ESI 16 (European Standard Investigator 16) and the PowerPlex^® ESI 17 Systems. The PowerPlex^® ESI 16 System combines the 11 loci compatible with the UK National DNA Database^®, contained within the AmpFlSTR^® SGM Plus^® PCR Amplification Kit, with five additional loci: D2S441, D10S1248, D22S1045, D1S1656 and D12S391. The multiplex was designed to reduce the amplicon size of the loci found in the AmpFlSTR^® SGM Plus^® kit. This design facilitates increased robustness and amplification success for the loci used in the national DNA databases created in many countries, when analyzing degraded DNA samples. The PowerPlex^® ESI 17 System amplifies the same loci as the PowerPlex^® ESI 16 System, but with the addition of a primer pair for the SE33 locus. Tests were designed to address the developmental validation guidelines issued by the Scientific Working Group on DNA Analysis Methods (SWGDAM), and those of the DNA Advisory Board (DAB). Samples processed include DNA mixtures, PCR reactions spiked with inhibitors, a sensitivity series, and 306 United Kingdom donor samples to determine concordance with data generated with the AmpFlSTR^® SGM Plus^® kit. Allele frequencies from 242 white Caucasian samples collected in the United Kingdom are also presented. The PowerPlex^® ESI 16 and ESI 17 Systems are robust and sensitive tools, suitable for the analysis of forensic DNA samples. Full profiles were routinely observed with 62.5 pg of a fully heterozygous single source DNA template. This high level of sensitivity was found to impact on mixture analyses, where 54–86% of unique minor contributor alleles were routinely observed in a 1:19 mixture ratio. Improved sensitivity combined with the robustness afforded by smaller amplicons has substantially improved the quantity of data obtained from degraded samples, and the improved chemistry confers exceptional tolerance to high levels of laboratory prepared inhibitors.