THE HUMORAL AND CELLULAR IMMUNE RESPONSES INDUCED BY HPV18L1-E6/E7 DNA VACCINES IN MICE

Objective To construct eukaryutic expression vector of HPV18 L1- E6, E7 chimeric gene and examine the humorul and cellular immune responses induced by this DNA vaccines in mice. Methods The C-terminal of major rapsid protein L1 gene and mutant zinc finger domains of early E6/7 oncogenes in HPV18 were integrated and inserted into eukaryotic expression vector pVAX1 to generate vaccines pVAX1-L1E6Mxx, ETMxx. CHO cells were transiently transfected with the individual construct. Target protein expressions in the lysate of the transfected cells were measured by ELISA and immunocytochemistry After BALB/c mice were vaccinated with various recombinant plusmids（pVAX1- L1-E6M3 or pVAX1-L1-E7M3 ） and immunie adjuvants （pLXHDmB7-2 or LTB） through different administration routes （intramuscular or intranasal） , the great cellular immune responses were produced us revealed by delayed-type hypersensitivity （DTH） and lymphocyte proliferation, and the expression of IL-4 and IFN- 7 cells in CD4^＋ and CD8^＋ subpopulations. Results The highly efficient expression of pVAX1-L1E6Mxx, E7Mxx vector in host eukaryotic cells were demonstrated both by ELISA and immunocytochemistry. The level of specific serum IgG against HPV in experiment groups mice was much higher than that of control group, and intranuscular immunization group had the highest antibody level. Intramuscular immunization groups were superior to intranasal immunization groups in DTH response, splenocyte proliferation and CD8^＋ IFN-γ^＋ cells number, but CD4^＋ IL4^＋ cell number was higher in intranasul immunization groups. The immunization groups using pLXHDmB7.-2 as adjuvant were superior to other groups in immunorespouse. Conclusion These DNA vaccines produce remarkable cellular and humorul immune responses in the mouse and may provide us prophylatic and therapeutic candidates for HPV induced cancer treatment.     

行政法意义上的非政府组织

非政府组织在行政管理学上有较多的研究,在行政法学上却少有论述,随着政治体制改革的深化,许多管理职能将由政府部门交由非政府组织行使,在这种情况下,我们必须从行政法的角度来研究权力转移的方式和条件,为这种权力转移提供法学依据.     

兔心房室隔过渡细胞的传导性及其电生理基础

目的　探索过渡细胞电生理特性与形态特征间的内在联系。方法　选 10只家兔做房室隔水平面及矢状面连续切片 ,HE染色观察。另 2 0只家兔采用标准玻璃微电极技术测定房室隔区正向传导速度。结果　房室结后方有两束过渡纤维分别延续至冠状窦口及其下方 ,分别为右后房结束和右下房结束 ,传导速度分别为 (0 .0 5 9± 0 .0 18)m·s-1和(0 .0 5 8± 0 .0 15 )m·s-1(P >0 .0 5 )。界嵴下段普通房肌传导速度为 (0 .2 0 3± 0 .0 4 0 )m·s-1。与前两者间均有显著性差异 (P <0 .0 1)。另外 ,过渡细胞动作电位有 4期自动除极 ,最大舒张电位 (- 6 3± 6 )mV ,普通房肌细胞无 4期自动除极 ,静息电位 (- 72± 4 )mV ,两者有显著性差异 (P <0 .0 5 )。结论　过渡细胞属慢反应自律细胞 ,传导速度比普通房肌慢。     

大骨节病和腺病毒感染的相关性研究

目的研究腺病毒感染与大骨节病(KBD)发病的关系。方法用巢式聚合酶链式反应(Nest-PCR)检测KBD病区和非病区儿童全血及KBD患者血清接种软骨细胞的腺病毒DNA,酶联免疫吸附实验(ELISA)检测KBD病区和非病区儿童血清腺病毒IgG抗体。结果KBD病区KBD儿童、病区对照组儿童和非病区儿童全血腺病毒DNA的阳性率分别为33.33%、34.92%和28.57%,腺病毒IgG抗体的阳性率分别为68.29%、65%和68.97%,差异无统计学意义(P>0.05);KBD患者血清培养细胞腺病毒DNA的PCR结果均为阴性。结论KBD病区和非病区儿童腺病毒均有较高的检出率;KBD的发病和腺病毒感染可能无直接病因学关系。     

糖尿病视网膜病变与血清VEGF含量的相关性

目的　探讨糖尿病 (DM)视网膜病变 (DR)程度与血清血管内皮生长因子 (VEGF)含量之间的关系。方法　 86例糖尿病患者按是否伴有DR及DR的严重程度分 3组 :糖尿病不伴DR(NDR)组 ,糖尿病伴单纯型DR(NPDR)组 ,糖尿病伴增殖型DR(PDR)组。采用定量双抗体夹心ELISA法检测血清VEGF含量 ,记录糖尿病病程。结果　 3组糖尿病患者血清VEGF水平逐组升高 ,NDR组为 (5 13.71± 16 8.92 )ng·L- 1 ;NPDR组为 (738.4 2± 337.4 2 )ng·L- 1 ;PDR组为 (95 4 .81± 2 6 1.2 0 )ng·L- 1 ,具有显著性差异 (P <0 .0 5 ) ,且血清VEGF含量与糖尿病病程显著正相关 (P <0 .0 0 0 1)。结论　糖尿病视网膜病变程度与血清VEGF含量直接正相关 ,血清VEGF含量测定可作为判定DR病变程度的参考指标     

基于最后计划者系统的工程项目进度计划管理

为了使工程项目进度计划更具有可操作性以及对进度的可控性,在编制项目进度计划过程中引入基于精益思想TFV理论的最后计划者系统。通过该系统的项目主计划、前瞻计划以及周计划的编制与实施,将进度计划的编制从传统的“推式”系统转变为“拉式”系统,可以实现项目计划的动态跟踪与管理。基于案例比较分析,证实该方法可以提高项目进度计划的有效性和可操作性。     

校园侵权行为类型表现及其责任形态的确定

校园侵权损害赔偿案件是近年来法院受理的各类侵权案件中常见的、多发的一种案件类型.学校、学生、第三人等不同侵权行为主体,在不同内容的侵权案件中,将面临不同侵权责任的承担,校园侵权行为类型表现的多样,直接影响到校园侵权责任形态的确定.本文围绕这一点,从校园侵权行为的概念和内涵入手,分析了校园侵权行为的具体类型,并对校园侵权责任形态的具体确定进行了理论探讨.     

2型糖尿病肾病患者血栓调节蛋白的变化

目的　探讨血浆内血栓调节蛋白 (TM)含量变化与 2型糖尿病肾病的关系及其在 2型糖尿病肾病发病中的意义 ,为早期诊断和治疗提供依据。方法　采用ELISA法 ,同时测定 6 9例 2型糖尿病患者和 12例正常人 (NC组 )血浆TM ;并根据 2 4h尿白蛋白排泄率 (UAER)将 2型糖尿病患者分为正常白蛋白尿组 (SDM组 ) ,微量白蛋白尿组 (即早期糖尿病肾病组 ,EDN组 ) ,大量白蛋白尿组 (即临床期糖尿病肾病组 ,CDN组 )。结果　①与NC组 (1.4 6 11± 0 .392 6 ) μg·L-1比较 ,血浆TM水平在SDM组 (2 .0 2 81± 0 .5 813) μg·L-1、EDN组 (2 .6 6 32± 1.0 0 3) μg·L-1、CDN组 (4 .5 86 1±2 .5 2 91) μg·L-1,均显著升高 (P <0 .0 0 5 ,P <0 .0 0 5 ,P <0 .0 0 1) ;与SDM组比较 ,EDN组和CDN组血浆TM显著升高 (P<0 .0 5 ,P <0 .0 0 1) ,CDN组血浆TM水平显著高于EDN组 (P <0 .0 0 1)。②血浆TM与UAER呈显著正相关 (r值为 0 .5 74 ,P <0 .0 0 1)。③血浆TM分别与病程、FPG、GHbAc、BUN、Cr、TC、LDL、SBP、DBP呈显著正相关 (r值分别为 0 .4 33、0 .2 2 7、0 .315、0 .2 2 8、0 .2 2 6、0 .2 0 7、0 .2 96、0 .4 77、0 .372 ,P均小于 0 .0 5 )。结论　血浆TM水平的测定有助于 2型糖尿病肾病的早期诊断     

铁路板式轨道结构平面有限元分析

无碴轨道是一种新型的轨道结构型式,越来越多地应用于高速铁路。目前,无碴轨道结构计算理论还不完善,多采用叠合梁的方法。现采用平面有限元方法,把荷载和轨道结构看成一个系统,对板式无碴轨道在荷载作用下的竖向位移和内力进行了分析,并通过MATLAB编程实现,计算结果符合无碴轨道结构基本原理。     

REAL-TIME DETECTION OF SURVIVIN mRNA EXPRESSION IN CERVICAL CANCER CELL LINES USING MOLECULAR BEACON IMAGING

Objective To detect the expression of survivin mRNA in cervical cancer cell lines using molecular beacon imaging technology. Methods Human cervical cancer cells （HeLa and SiHa） and human fetal lung fibroblast HFL-I were cultured in vitro. After adding 100 nmol/L survivin mRNA molecular beacon, the fluorescent signals were observed under fluorescent microscope. The expressions of survivin in cervical cancer cells and HFL-I cell were examined by immunocytochemical streptravidin-biothin peroxidase （SP） assay at the same time. Results Two kinds of survivin mRNA molecular beacon, with different color fluorescence, had strong fluorescent signal in cervical cancer cell lines, and the signal in SiHa cell line was stronger, but these signals were not found in HFL-I ; Immunocytochemical staining of positive survivin was located in the cytoplasm of cervical cancer cell lines HeLa and SiHa, whereas, no expression of survivin was detected in HFL-I cell line. Conclusion The technology of molecular beacon imaging can be used to detect the expression of survivin mRNA in viable cells successfully, and may provide a new approach to the diagnosis of early stage cervical cancer and the following-up in the clinic.