基于MSP430动态心电存储系统的设计

目的：解决动态心电图存储容量有限、波形不能实时显示的不便。方法：设计一种基于TI公司超低功耗混合信号处理器MSP430F149的动态心电数据实时存储、传输系统。结果：此系统由MSP430F149单片机、心电数据采集、LCD显示、SD卡存储、USB数据传输等部分组成,它能完成心电数据的实时采集、显示、大容量存储,以及心电数据的USB传输。结论：经临床初步实验,该仪器具有较强的抗干扰能力、性能稳定、功耗低、携带方便,应用前景广阔。     

子宫内膜癌组织中FHIT基因启动子区域CpG岛甲基化状况的研究

目的 :检测子宫内膜癌组织中FHIT基因启动子区域CpG岛的甲基化状况 ,分析其与临床病理特征之间的关系 ,探讨FHIT基因启动子区域CpG岛的甲基化在子宫内膜癌发生发展中的作用。方法 :采用甲基化特异的PCR方法对亚硫酸氢盐修饰过的 35例子宫内膜癌组织、癌旁组织及患者自身外周血白细胞DNA和 2 0例非癌患者子宫内膜组织DNA的FHIT基因启动子区域CpG岛的甲基化状况进行分析。结果 :癌组织中FHIT基因启动子区域CpG岛的甲基化率为 2 5 .71 % ,癌旁组织和外周血中也存在相似的甲基化率。非癌患者的子宫内膜组织中FHIT基因的CpG岛无甲基化。癌组织与非癌患者子宫内膜组织的甲基化率之间的差异有统计学意义 (P <0 .0 5 )。DNA甲基化与临床病理特征之间无相关性。FHIT基因启动子区域CpG岛的甲基化为子宫内膜癌发生中的早期事件。结论 :子宫内膜癌组织中存在FHIT基因启动子区域CpG岛的甲基化 ,CpG岛的甲基化与临床病理特征之间无相关性。DNA甲基化为子宫内膜癌发生中的早期事件。癌旁组织和外周血可用于癌相关基因启动子区域CpG岛甲基化的检测     

A phase 2b randomized,controlled trial of the efficacy of the GMZ2 malaria vaccine in African children

BackgroundGMZ2 is a recombinant protein malaria vaccine, comprising two blood-stage antigens of Plasmodium falciparum, glutamate-rich protein and merozoite surface protein 3. We assessed efficacy of GMZ2 in children in Burkina Faso, Gabon, Ghana and Uganda.MethodsChildren 12–60 months old were randomized to receive three injections of either 100 μg GMZ2 adjuvanted with aluminum hydroxide or a control vaccine (rabies) four weeks apart and were followed up for six months to measure the incidence of malaria defined as fever or history of fever and a parasite density ⩾5000/μL.ResultsA cohort of 1849 children were randomized, 1735 received three doses of vaccine (868 GMZ2, 867 control-vaccine). There were 641 malaria episodes in the GMZ2/Alum group and 720 in the control group. In the ATP analysis, vaccine efficacy (VE), adjusted for age and site was 14% (95% confidence interval [CI]: 3.6%, 23%, p-value = 0.009). In the ITT analysis, age-adjusted VE was 11.3% (95% CI 2.5%, 19%, p-value = 0.013). VE was higher in older children. In GMZ2-vaccinated children, the incidence of malaria decreased with increasing vaccine-induced anti-GMZ2 IgG concentration. There were 32 cases of severe malaria (18 in the rabies vaccine group and 14 in the GMZ2 group), VE 27% (95% CI −44%, 63%).ConclusionsGMZ2 is the first blood-stage malaria vaccine to be evaluated in a large multicenter trial. GMZ2 was well tolerated and immunogenic, and reduced the incidence of malaria, but efficacy would need to be substantially improved, using a more immunogenic formulation, for the vaccine to have a public health role.     

非霍奇金淋巴瘤p73基因甲基化及去甲基化的研究

目的探讨p73基因异常甲基化在非霍奇金淋巴瘤（non—Hodgkin’s lymphomas,NHL）中的分布,去甲基化p73基因的再表达。方法采用甲基化特异性PCR（Methylation specific polymerase chain reaction,MSP）方法检测22例非霍奇金淋巴瘤p73基因CpG岛甲基化状态。使用反转录PCR（Reverse transoription polymerase chain reaction,RT-PCR）方法检测CdR药物作用前后P73基因mRNA的表达情况。结果非霍奇金淋巴瘤p73基因甲基化的发生率为27.3％（6／22）,高度恶性比低度恶性更易发生甲基化,其发生率分别为42．8％和0％,5-杂氮-2’．脱氧胞嘧啶（5-aza-2’-deoxycytidine,CdR）在4．0～8．0μmol/L时可诱导非霍奇金淋巴瘤细胞p73去甲基化。结论p73基因甲基化可能是非霍奇金淋巴瘤发病的原因之一,去甲基化治疗是可行的。     

肺癌患者血浆中APC基因启动子甲基化定量检测研究

背景与目的：APC基因编码的蛋白参与信号转导途径,大量研究证实APC启动子区的高甲基化在肿瘤的发生发展中起重要作用。本研究建立血浆APC基因实时荧光定量甲基化特异性基因扩增（methylation—specific PCR,MSP）检测方法,并对临床肺癌患者血浆进行检测,以确定该方法在肺癌诊断中的应用价值。方法：将APC基因启动子甲基化阳性肺癌细胞株NCI—H460细胞用有限稀释法获取单个集落形成的细胞,以经典的酚一氯仿法提取细胞DNA,并用MSP对APC基因甲基化进行验证,紫外分光光度计定量并以10倍稀释的浓度梯度依次投入到200μL健康人血浆中．得到模拟肺癌患者血浆,利用磁珠核酸提取方法从模拟肺癌及肺癌患者、肺部良性疾病患者和健康对照者血浆中提取DNA,对血浆DNA模板进行亚硫酸氢盐化学修饰;以模拟血浆样品作为标准品,采用外标准曲线法对各种血浆样品中APC甲基化进行定量分析。结果：所建立的实时荧光定量MSP检测方法的线性范围为1．5×10^2-1．5×10^5拷贝／mL。用该方法检测甲基化肿瘤细胞的DNA其最低检测限为1．5×10^2拷贝／mL。78例肺癌患者有40例组织中检测出APC基因甲基化阳性．其中的19例（47．5％）肺癌患者血浆中APC基因启动子甲基化阳性,APC甲基化浓度为1．67×10^2-6．78×10^3拷贝／mL,中位浓度为1．60×10^3拷贝／mL。38例组织阴性的肺癌患者、31例肺部良性疾病患者和23例健康者的血浆APC基因启动子甲基化均为阴性。结论：实时荧光定量MSP检测血浆APC基因甲基化在肺癌诊断方面具有潜在应用价值。     

Promoter hypermethylation of the ADAM23 gene in colorectal cancer cell lines and cancer tissues

Promoter hypermethylation of the ADAM23 gene, which is normally involved in cell‐to‐cell and cell‐to matrix adhesion, has been reported in pancreatic, breast and brain cancer, and recently the role of this gene was examined in gastric cancer. In this study, we analyzed ADAM23 expression in colorectal cancer cell lines and examined its methylation by methylation‐specific PCR (MSP) and bisulfate‐modified DNA sequencing analysis. Methylated cells were treated with 5‐aza‐2′‐deoxycytidine to restore the ADAM23 expression. We then examined ADAM23 methylation status in colorectal cancer tissues and their corresponding normal tissues. We found that ADAM23 was aberrantly silenced or expressed at very low levels in 28 of the 32 (88%) colorectal cancer cell lines. MSP analysis showed that ADAM23 was methylated in 29 of 32 (91%) colorectal cancer cell lines and attenuated expression of ADAM23 was found to be related to hypermethylation in its promoter region. Moreover, the CpG dinucleotide methylation threshold of 70–90% was found to be required for complete silencing. In addition, when some cell lines without ADAM23 expression were treated with 5‐aza‐2′‐deoxycytidine, ADAM23 was reexpressed. In colorectal cancer tissues, the promoter region of ADAM23 was hypermethylated in 36 of 76 (47%). These results demonstrated that ADAM23 may be down‐regulated by aberrant promoter hypermethylation during the progression of colorectal cancer. © 2008 Wiley‐Liss, Inc.     

恶性疟原虫裂殖子表面蛋白-2DNA疫苗诱导小鼠T淋巴细胞的增殖

目的　为了探讨恶性疟原虫FCC - 1/HN株裂殖子表面蛋白 - 2 (MSP - 2 )DNA疫苗在小鼠体内诱导的免疫应答的特性及抗感染的保护性免疫机制。方法　将重组真核表达质粒pBK/MSP - 2经骨骼肌注射BALB/c小鼠 ,小鼠经DNA疫苗免疫 8w后 ,用流式细胞仪分析脾脏T淋巴细胞的分化 ,并体外培养脾脏细胞 ,用夹心ELISA法测定IFN -γ和IL - 2的产生。结果　与对照组相比 ,疫苗组CD+ 4CD8+ T淋巴细胞有显著性的增高 ,体外培养的脾脏细胞IFN -γ有一个高浓度的分泌。结论　恶性疟原虫FCC - 1/HNMSP - 2DNA疫苗诱导了一个TH1的免疫应答类型     

乳腺癌雌激素受体α基因启动子甲基化与其蛋白表达的关系

目的：检测乳腺癌雌激素受体α（estrogen receptor α,ERα）基因启动子区CpG岛甲基化状态,探讨乳腺癌ERα启动子甲基化与其蛋白表达及临床病理的关系。方法：采用免疫组化EnVisionTM二步法检测20例正常乳腺组织、44例乳腺癌组织ERα基因的表达,同时运用甲基化特异性PCR（Methylation-Specific PCR,MSP）及测序法分别检测ERα基因启动子A区和B区CpG岛的甲基化状态。结果：32例ERα阳性乳腺癌组织中,ERα启动子A区、B区甲基化率分别为28.2%和18.8%。12例ERα阴性乳腺癌组织中,ERα启动子A区、B区甲基化率分别为66.7%和58.3%。ERα阳性乳腺癌组、ERα阴性乳腺癌组、乳腺癌组的ERα启动子A、B区甲基化率均较正常组显著增高（P〈0.05）;ERα阴性乳腺癌组的ERα启动子A、B区甲基化率较ERα阳性乳腺癌组均显著增高（P〈0.05）。乳腺癌组织ERα表达与启动子区甲基化之间呈显著负相关。结论：乳腺癌组织ERα基因表达沉默与其启动子A区、B区CpG岛甲基化相关,后者有可能成为乳腺癌预后不良的分子检测指标。     

乳腺癌细胞中PNRC基因启动子区CpG岛甲基化状态的研究

目的 研究乳腺癌细胞中富含脯氨酸核受体辅活化子(proline-rich nuclear receptor coactivator protein,PNRC)基因启动子CpG岛的甲基化状态及在PNRC转录调节中的作用.方法 应用PT-PCR检测PNRC基因在正常乳腺细胞与乳腺癌细胞中的表达情况,并运用"MethPrimer"软件对PNRC启动子区进行分析,预测CpG岛,通过甲基化特异性PCR(methylation-specific PCR,MSP)检测PNRC启动子区CpG岛的甲基化状况;PT-PCR及Northern杂交检测乳腺癌细胞经甲基转移酶抑制剂5-氮杂-2'-脱氧胞苷(5-aza-2'-deoxycytidine,5-Aza-dc)作用后,PNRC基因mRNA的表达情况;Westernblot检测5-Aza-dc干预对PNRC蛋白表达的影响;将含PNRC启动子序列的报告质粒转染乳腺癌细胞,再经5-Aza-dc处理后,检测PNRC基因启动子的活性.结果 PNRC基因在乳腺癌细胞中的表达较正常乳腺细胞明显降低;乳腺癌细胞PNRC基因启动子区存在CpG岛甲基化;乳腺癌细胞经5-Aza-dc处理后,PNRC基因mRNA及蛋白表达水平显著增加,其PNRC基因启动子的活性也明显增强.结论 PNRC基因启动子区的高甲基化导致PNRC在乳腺癌细胞中表达降低.     

Combined Effect of Metastasis-Related MicroRNA,miR-34 and miR-124 Family,Methylation on Prognosis of Non–Small-Cell Lung Cancer

Background

Many patients with non–small-cell lung cancer (NSCLC) still develop tumor metastasis and recurrence after pulmonary resection and are the primary causes of lung cancer treatment failure and death. MicroRNAs (miRs) have central roles during tumor metastasis and many miR genes are potentially subjected to control by DNA methylation in multiple tumor types. Recently, miR-34 and miR-124 have been demonstrated as potential regulators of the metastasis process in several cancer types.