人乳头瘤病毒11型L1/CD80嵌合真核表达质粒的构建和鉴定

生殖道尖锐湿疣主要是由人乳头瘤病毒6型(HPV6)和11型(HPV11)感染引起。HPV病毒仅存在终末分化的复层鳞状上皮组织中，含量很少。HPV病毒的晚期表达基因L1所表达的外壳蛋白不仅是主要的外壳蛋白，还是主要的病毒抗原蛋白，在宿主细胞表面有其蛋白受体，可成为预防性疫苗最理想的靶抗原。我们构建了含有完整的HPV11型L1基因的真核表达质粒，为增强其作为治疗性疫苗的免疫效果，并进一步将HPV11L1基因和共刺激分子CD80进行连接重组，构建HPV11 L17／CD80嵌合基因的DNA疫苗，为下一步研究其免疫原性提供试验数据。     

巨噬细胞移动抑制因子在妇科恶性肿瘤中的表达

目的观察妇科恶性肿瘤患者血清、腹腔液中巨噬细胞移动抑制因子(MIF)的表达及其与恶性肿瘤的相关性。方法选择术前未行任何治疗的妇科恶性肿瘤患者46例作为实验组(卵巢癌21例,子宫恶性肿瘤25例),良性肿瘤患者24例为对照组。ELISA法检测术前肘静脉血、术中腹腔液的MIF浓度。同时测定血清血管内皮生长因子(VEGF)及CA125浓度。结果实验组腹腔液MIF的浓度(21.98±11.27 ng/ml)与对照组(13.85±8.11 ng/ml)相比差异显著(P<0.01);而血清MIF浓度(分别为5.13±3.53 ng/ml和3.42±2.06 ng/ml)无明显差异(P>0.05)。卵巢癌患者腹腔液和血MIF的浓度(26.26±11.46 ng/ml,5.52±3.04 ng/ml)分别与良性卵巢肿瘤(11.10±6.68 ng/ml,2.92±1.90 ng/ml)相比,均有显著差异(P<0.001,P<0.01);腹腔液与血清中MIF浓度有相关(r=0.5821,P<0.01);腹腔液MIF浓度与血CA125有关联(r=0.6649,P<0.05),与血清VEGF也相关(r=0.405,P<0.05)。子宫恶性肿瘤患者腹腔液和血清MIF的浓度(18.20±8.67 ng/ml和4.68±3.24 ng/ml)与子宫肌瘤(17.24±6.32 ng/ml和2.8±2.20 ng/ml)相比,均无差异(均P>0.05)。结论卵巢癌患者腹腔液与血清均有MIF表达,而腹腔液MIF呈高表达,可能与CA125类似,主要源于肿瘤组织。MIF与VEGF可能存在互相诱导分泌的关系。     

实时荧光PCR技术定量检测乳腺癌患者血浆循环DNA

目的建立一种实时荧光定量PCR方法,并用该方法检测乳腺癌患者血浆循环DNA水平。方法用QIAamp血液DNA抽提试剂盒提取61例乳腺癌患者和25例健康妇女血浆循环DNA,用实时荧光定量PCR技术(SYBR Green I染料法)进行定量,结果用设定临界域值时的循环数的值(Ct值)表示,并对定量结果进行ROC曲线分析。结果:乳腺癌患者Ct值(27.71±1.41)显著低于健康对照(30.37±1.32,P<0.001),说明乳腺癌患者血浆循环DNA水平显著高于健康对照组;ROC曲线下面积为0.921。结论应用实时PCR技术检测乳腺癌患者血浆循环DNA水平有可能成为一种无创性乳腺癌分子诊断方法。     

人乳头瘤病毒11型L1/E7嵌合DNA疫苗的构建及其诱导的免疫效应

Objective To construct chimerical DNA vaccine plasmid of human papiUomavirus type 11 (HPV11) L1-E7, and to evaluate its immunogenicity. Methods Molecular cloning techniques were used to construct recombinant plasmid PeDNA3 L1-E7 as a DNA vaccine. BALB/c mice were vaccinated with DNA recombinants through muscle injection. IL-2 and γ-INF secreted by immunized spleens lymphocyte and HPV 11 LI or E7 specific antibodies were assayed by ELISA method. Spleens lymphocyte proliferation was measured by MTT assay. Results The chimerical DNA plasmid of pcDNA3 LI-E7 was constructed correctly. Specific anti-HPV11 E7 and L1 antibodies, specific lymphocyte proliferation and secretions of IL-2 and γ-INF were detected in vaccinated mice. Conclusion Specific immune response, including cellular immunity and humoral immunity, could been detected in mice vaccinated with chimerical DNA vaccine of pcDNA3 L1-E7.     

人乳头瘤病毒11型E7与共刺激分子CD80嵌合DNA疫苗质粒的构建及鉴定

目的:构建人乳头瘤病毒11型(HPV ll)E7与共刺激分子CDS0嵌合DNA疫苗质粒。方法:从尖锐湿疣病变组织中提取DNA制备模板,采用聚合酶链反应(PCR)技术扩增HPV ll-E7基因,经双酶切后定向克隆到pcDNA3.1(+)中,获得pcDNA3.1(+)/HPV llE7,测序鉴定后,再用PCR方法扩增去除终止密码子的E7基因片段,按上述方法插入质粒pcD-NA3.1(+)/CD80中CD80上游,构建pcDNA3.1(+)/HPV llE7-CD80融合基因真核表达质粒。结果:去除终止子的E7基因片段成功正向插入CD80上游,双向测序分析显示序列无误,pcDNA3.1(+)/HPV llE7-CD80嵌合真核表达质粒构建成功。结论:HPV ll-E7/CD80嵌合DNA疫苗质粒构建成功,为研究该疫苗在尖锐湿疣防治中的作用奠定了基础。     

人乳头瘤病毒11型L1/E7嵌合DNA疫苗的构建及其诱导的免疫效应

Objective To construct chimerical DNA vaccine plasmid of human papiUomavirus type 11 (HPV11) L1-E7, and to evaluate its immunogenicity. Methods Molecular cloning techniques were used to construct recombinant plasmid PeDNA3 L1-E7 as a DNA vaccine. BALB/c mice were vaccinated with DNA recombinants through muscle injection. IL-2 and γ-INF secreted by immunized spleens lymphocyte and HPV 11 LI or E7 specific antibodies were assayed by ELISA method. Spleens lymphocyte proliferation was measured by MTT assay. Results The chimerical DNA plasmid of pcDNA3 LI-E7 was constructed correctly. Specific anti-HPV11 E7 and L1 antibodies, specific lymphocyte proliferation and secretions of IL-2 and γ-INF were detected in vaccinated mice. Conclusion Specific immune response, including cellular immunity and humoral immunity, could been detected in mice vaccinated with chimerical DNA vaccine of pcDNA3 L1-E7.     

胃癌组织中胸苷酸合成酶mRNA表达与预后的关系

目的探讨胃癌组织中胸苷酸合成酶（TS）mRNA表达水平与预后的关系。方法以G6PDH作内参照，采用实时定量RT-PCR技术检测41例胃癌组织TS mRNA表达水平。结果胃癌组织TS mRNA表达水平的中位数为0．93。TS高表达组（〉0．93）和低表达组（≤0．93）之间无瘤生存期和总生存期差异有显著性（P〈0．05）；而TS mRNA表达与年龄、性别、淋巴结转移、组织学分级及临床分期均无相关性（P〉0．05）。结论检测TS mRNA表达水平对判断胃癌病人预后有很好的预测价值。     

人乳头瘤病毒11型L1/E7嵌合DNA疫苗的构建及其诱导的免疫效应

Objective To construct chimerical DNA vaccine plasmid of human papiUomavirus type 11 (HPV11) L1-E7, and to evaluate its immunogenicity. Methods Molecular cloning techniques were used to construct recombinant plasmid PeDNA3 L1-E7 as a DNA vaccine. BALB/c mice were vaccinated with DNA recombinants through muscle injection. IL-2 and γ-INF secreted by immunized spleens lymphocyte and HPV 11 LI or E7 specific antibodies were assayed by ELISA method. Spleens lymphocyte proliferation was measured by MTT assay. Results The chimerical DNA plasmid of pcDNA3 LI-E7 was constructed correctly. Specific anti-HPV11 E7 and L1 antibodies, specific lymphocyte proliferation and secretions of IL-2 and γ-INF were detected in vaccinated mice. Conclusion Specific immune response, including cellular immunity and humoral immunity, could been detected in mice vaccinated with chimerical DNA vaccine of pcDNA3 L1-E7.     

粪便DNA甲基化分析在结直肠癌早期诊断中的价值

近年来,研究表明DNA甲基化改变与肿瘤的发生发展关系密切,有望用于恶性肿瘤的分子诊断。由于结直肠癌(CRC)组织持续脱落细胞到粪便中,因此检测粪便中CRC相关基因的甲基化状态有可能成为一条无创诊断CRC的新途径。增生性息肉蛋白基因(HPPI)是在结肠增生性息肉中     

乳腺癌患者血浆D-二聚体的检测及其临床意义

目的：检测乳腺癌患者手术前后血浆D-二聚体水平，并探讨其与其它乳腺癌临床参数的关系。方法：用ELISA方法检测51例乳腺癌妇女血浆D-二聚体水平，以10例良性乳腺肿瘤女性患者和42例健康人作为对照。结果：乳腺癌患者血浆D-二聚体水平显著高于对照组，且随着肿瘤发展转移而升高；雌、孕激素受体表达均达为阴性的患者DD水平明显较雌激素和/或孕激素表达职性者高。结论：检测乳腺癌患者血浆D-二聚体对于病情的诊断，分期及预后具有重要意义。