Ig/Ig-TCIC的免疫生物学

实验以抗人m、g、a链的羊IgG作为捕捉抗体，分别以HRP-抗人g链和a链的羊IgG、HRP-抗人m链和a链的羊IgG、HRP-抗人m链和g链的羊IgG为检测抗体，建立了检测6种不同类型Ig/Ig-TCIC（IgM/IgG、IgG/IgA、IgA/IgM、IgG/IgM、IgA/IgG、IgM/IgA-TCIC）的捕捉法ELISA。经阻断实验、替代实验、以及板内、板间和批间变异考察，证实该方法特异性高、重复性好。在此基础上，较系统地研究了各类Ig/Ig-TCIC，从而对Ig/Ig-TCIC的免疫生物学意义有较为全面的认识。 在研究中，着重于健康人和患者、传染病和非传染性疾病、不同疾病以及...Using goat IgG anti-human-μ,γ,α chain as a capture antibody and goat IgG HRP-anti- human-γ,αchain, goat IgG HRP-anti- human-μ,αchain, and goat IgG HRP-anti- human-μ,γ chain as detecting antibodies, respectively, six types of capture ELISA methods had been established to detect Ig/Ig-TCIC (IgM/IgG、IgG/IgA、IgA/IgM、IgG/IgM、IgA/IgG、and IgM/IgA). They have been proven to be high specific and good repr...学位：理学硕士院系专业：生命科学学院生物学系_细胞生物学学号：19972602

Basic Study on Application of Gene Carrier in Gene Therapeutic Drugs

根据自主知识产权的专利工艺路线，合成具有分支结构、分子量30kD的阳离子聚合物，以此为基础构建sofast基因载体，用于大分子核酸的体外和体内的基因传递。通过sofast/DNA复合物形成、粒径、zeta电位、抗核酸酶能力、酸碱缓冲能力、结合核酸能力，以及基因融合肽diINF-7和工作介质盐浓度的影响等分析，对sofast基因载体的转染机制做了初步的探讨。在体外试验中，采用sofast基因载体将质粒pEGFPN1、pCMV-lacZ和pGL3分别转染到HEK297细胞，通过所表达的绿色荧光蛋白、b-半乳糖苷酶和荧光素酶的检测，证实其具有高转染效率；通过HUV-EC等众多细胞株的转染，结果表明其...According to the independent intellectual property patent process route，a branched and 30KD cationic polymer was synthesized, which the sofast gene carrier was constructed based on. The sofast gene carrier can be applied as big molecular nucleic acids carrier both in vitro and in vivo. In this paper we investigated its transfection mechanism as a gene carrier through formation of sofast/DNA comple...学位：理学博士院系专业：生命科学学院生物化学与生物技术系_生物化学与分子生物学学号：B20032602

Study on Quantitative Method and Immune Regulation of Ig/Ig two-component-Determined Circulating Immune Complexes

目的　建立Ig/Ig -TCIC的定量方法 ,研究Ig/Ig -TCIC是否具有免疫调节作用。 方法　以Ig的免疫生物学性质和数学计算相结合进行定量。通过研究Ig/Ig-TCIC与C3/Ig -TCIC、全血清和复合Ig与补体等体液免疫指标的关系来探讨其免疫调节作用。结果　成功地建立了Ig/Ig -TCIC的定量方法 ,并发现Ig/Ig -TCIC与有关体液免疫指标的相互关系具有明显规律性。结论　本定量法简便实用。Ig/Ig -TCIC作为免疫网络的中间产物在免疫调节中起重要作用Objective To establish a quantitative method to Ig/Ig two component determined circulating imune complexes(Ig/Ig-TCIC) and study whether Ig/Ig-TCIC was related to the function of immune regulation.Methods Quantity of Ig/Ig-TCIC was carried out by the combination of Ig immune biology with mathematics.The immune regulation of Ig/Ig-TCIC was characterized by the correlation of Ig/Ig-TCIC with C3/Ig-TCIC,serum Ig and C3,and complexed Ig and C3.Results The quantitative method was established and significant regularity was found from the association of Ig/Ig-TCIC with these humoral immune markers.Conclusion This quantitative method was simple and useful.Ig/Ig-TCIC plays an important role in immune regulation as a middle product of immune network.教育部优秀骨干教师基金;; 福建省自然科学基金资助项目(C970 16

In┐depthStudyofSerumImmunoglobulininCancerPatientsHuangXueFangWangSanyingPengXuanxianetalTheStateLaboratoryForTumorCelEnginering,XiamenUniversity,XiamenAbstractInordertostudythechangesinimmunoglobulinIgpaternsincancerpa-tients,serumIg(totalIg)wascategori

目的：评价恶性肿瘤患者的免疫球蛋白反应类型。方法：将血清免疫球蛋白（Ig）剖解为游离和复合两部分进行研究。以食管癌、喉癌、胃癌、乳腺癌、子宫癌、鼻咽癌、卵巢癌和非霍奇金淋巴瘤（nHl）与健康者分别为实验组和对照组。结果：患者游离Ig的变化与总Ig一致，而复合Ig则与之不同，对其的检测更有利于对患者免疫状况的评价。从而说明，既往有关血清Ig含量的测定，主要反映游离Ig部分的免疫反应状况。结论：免疫反应产物的剖解分析可以更进一步认识免疫反应本质。在对肿瘤患者Ig比例的研究中，发现肿瘤患者的复合Igg普遍严重下降。从而认为特异性Igg水平的明显下降是患者免疫功能降低的重要原因，并提出深入研究其机理，将可能为探索肿瘤发病机理和治疗找到一条新路In order to study the changes in immunoglobulin Ig patterns in cancer patients, serum Ig (total Ig) was categoried into Free Ig and complex Ig For analysis in patient with cancers of esophagus,larynx,stomach, breast, nasopharynx, ovary and non Hodgkin's lymphomas as well as in healthy individuals as control.It was Found that the variations of Free Ig were in good mimicry with those of the total Ig,whereas complex Ig had its characteristic Features paralleling the immune status of the patients.It was thought, thereFore, previous reports on serum Ig mainly concentrated in the immune responses of Free Ig,and that in depth analysis of immune elements could only enable unveiling the truth .Furthermore, we Found that complex IgG was markedly decreased in almost all of these malignancies in terms of Ig ratios.It is suggested that the marked decrease of speciFic IgG levels might be the key reason For declining immune ability of cancer patients and that Further study of its mechanism would be possible to Find a new approach For exploring tumor pathogenesis and therapy.福建省自然科学基

潜在血源传播患者梅毒血清学TRUST/TPPA与IgM抗体联合检测

目的为潜在血源传播患者筛选梅毒血清学传染病指标。方法采用回顾性研究,对2006年6月1日～2009年12月31日本院23720例潜在血源传播患者与门诊患者的TRUST/TPPA联合检测结果进行分析比较;分析不同年度潜在血源传播患者TRUST-/TPPA+的百分率变化;对其中118份TRUST-/TPPA+的潜在血源传播患者血清标本同时采用TPPA法和FTA-ABS法进行Tp-IgM抗体检测。结果潜在血源传播患者和门诊患者的梅毒抗体阳性率分别为8.62%和26.69%,二者差异有显著性;在梅毒抗体阳性标本中,TRUST-/TPPA+占有相当的比例,二者分别为3.12%和7.50%;潜在血源传播患者中TRUST-/TPPA+的百分率呈逐年上升趋势;118份TRUST-/TPPA+的潜在血源传播患者血清,经TPPA法和FTA-ABS法检出TP-IgM抗体阳性分别为13例和10例。结论潜在血源传播患者的梅毒传染性指标的监测应至少包含特异性抗体试验,如TPPA;TP-IgM检测有助于判断其传染性

HBVDNA/补体双特异性免疫复合物的检测

以羊抗人C1Q和b因子Igg为捕捉抗体，分别用以捕捉通过经典和旁路途径激活补体的dAnE颗粒抗原抗体免疫复合物中的C1Q和b因子，再利用引物的特异性和PCr的强大扩增能力对捕捉物中的HbVdnA进行检测，成功地建立了检测HbVdnA/补体双特异性循环...国家自然科学基

潜在血源传播梅毒患者血清抵抗特性分析

目的研究潜在血源传播梅毒患者血清抵抗特性,探讨其传染性和复发的血清学指标。方法在回顾性分析2005年6月-2009年5月厦门大学附属中山医院收治的714例潜在血源传播梅毒患者的血清抵抗特性基础上,采用荧光密螺旋体抗体吸收试验(FTA-ABS)进行梅毒特异性IgM抗体(TP-IgM)检测。结果 714例潜在血源传播梅毒患者中,有242例患者经1年梅毒规范治疗后,其反应素仍保持阳性,血清抵抗率33.89%,其中男性患者血清抵抗率28.08%,显著低于女性患者(40.54%),差异有统计学意义(χ2=12.306,P<0.05);不同检测年度、不同年龄患者血清抵抗率差异无统计学意义;采用FTA-ABS法,242例血清抵抗患者中检出TP-IgM的阳性率为32.23%,显著高于梅毒血清治愈组(10.70%)和献血员组(0)。结论潜在血源传播梅毒患者有着较高的血清抵抗率,是梅毒治疗失败的重要表现,并容易复发和具有传染性,TP-IgM可作为其复发和具有传染性的血清学指标

Immuno capture PCR for HBVDNA/Ig-two-component-determined circulating immune complexes

目的:建立一种免疫捕捉法PCR技术,研究血清免疫复合物中不同Ig类型抗体结合HBV的情况,并初步探讨其意义。方法:以羊抗人μ、γ、α链的IgG为捕捉抗体,再利用PCR扩增捕捉物中的HBVDNA。结果:成功地建立了检测HBVDNA/IgM、IgG和IgATCIC的免疫捕捉法PCR技术。该技术特异性强、敏感性高、重复性好,可以显著提高HBVDNA的检出率。同时发现,结合HBV的抗体的Ig类型组合有明显差异,三者均阳性最高。结论:说明乙肝患者血清免疫复合物中有较多的病毒颗粒。研究HBVDNA/IgTCIC对乙肝发病机理的深入阐明可能具有重要意义。Objective: To establish an immuno capture PCR for the study of classes of immunoglobulins complexed with HBV and of their significance.Methods:Using goat IgG to human μ、γ、α chain as capture antibodies and prime specific PCR as a detecting method.Results:An immuno capture PCR had been established to detect HBVDNA/Ig TCIC (HBVDNA/IgM TCTC,HBVDNA/IgG TCIC, HBVDNA/IgA TCIC). This method was specific,sensitive and reproducible,by which more patients with positive HBVDNA could be found. It was also characted that there were significantly different positive rates among the types of combinations of IgM,IgG and IgA, in which the group with the three of them was hightest.Conclusion:Conclusion:These findings suggested that more HBV may be hide within circulating immune complexes and the study of HBV DNA /Ig TCIC may play an important role in more understanding of pathogenesis in patients with hepatitis B.国家自然科学基金!3 96 6 0 0 7

Determination of the potency of antibiotic G-418 by microbial turbidimetry

采用微生物比浊法检定抗生素G-418的效价。在本实验的浓度范围内(3.76μg/ml～8.77μg/ml),标准曲线线性方程为Y=-0.9184X+0.9917,回归方程为Y=-0.9215X+0.9927,r=-1.0002,其浓度与吸收度的线性关系良好,标准曲线成立。样品检测可靠性测验,回归显著(P<0.05),S、T为平行直线,实验结果可靠。采用微生物比浊法测定G-418的效价,具有耗时短,灵敏度高,操作简便,不需要特殊设备,且人为影响因素少等诸多特点,值得推广应用。A microbial turbidimetry was used to determine the potency of antibiotic G-418.The calibration curve was inear in the range of 3.76U/ml～8.77U/ml, while the calibration curve linear equation is Y= -0.9184X+0.9917 and the regression equation is Y=-0.9215X+0.9927,r=-1.0002. The reliability of the method was discussed as well. The result is reliable with significant regression (P<0.05) and S and T parallel lines. The result indicates that the method of microbial turbidimetry can be used more widely because of its convenience, accuracy and quickness.科技部科技创新基金资助(04C26213510157

Detection and Their Significance of Ig/Ig Two component determined Circulating Immune Complexes United by Different Classes of Ig

为研究独特型免疫网络的中间产物，以抗人IgM、Igg、IgA 分别为包被抗体，以异类酶标抗人Ig 为检测抗体，建立了检测由异类Ig 形成的Ig/Ig 双特异性循环免疫复合物（TWO - COMPOnEnT -dETErMInEd CIrCulATIng IM MunE COMPlEXES，Ig/Ig - TCIC） 的ElISA 技术。采用该技术对健康人群进行检测，并按年龄组和性别组进行分析。结果发现，同一TCIC 的年龄组含量变化趋势依包被和酶标抗体不同而有明显差异，而不同TCIC 的年龄组含量变化趋势因酶标抗体相同而基本相同；TCIC 含量女性明显高于男性。提示，在检测免疫复合物的ElISA 中，所得结果与所用何种酶标抗体以及复合物的分子比有关。深入研究Ig/Ig - TCIC 对阐明免疫网络中间产物的免疫调节作用具有重要意义。In order to study middle products of immune network, ELISA methods had been established to detect Ig/Ig two component determined circulating immune complexes (Ig/Ig TCIC) united by different classes of Ig, in which anti human μ、γ or α chain specific IgG was used as a coating antibody and HRP labeling antibody different to the coating Ig was used as a detecting one.The distributions of Ig/Ig TCIC were characterized in age and sex groups of healthy individuals.It was found that there were significant differences between the changes in level of a same Ig/Ig TCIC detected by cross coating and label ing antibodies, but similarity between different Ig/Ig TCIC detected by a equally labeling antibody.Foreover, it was found that the amounts of Ig/Ig TCIC were significant higher in woman group than in man one.These finding suggest that in ELISA tests to immune complexes, the results are associated with a selected labeling antibody and ratios of detected materials.And it is important to further study the Ig/Ig TCIC middle products in immune network for understanding of their functions of immune regulation.教育部重点资助;福建省自然科学基