Nicotinamide N-methyltransferase protein expression in renal cell cancer

Objective: To understand the function of nicotinamide N-methyltransferase (NNMT) protein as tumor biomarker in renal carcinoma. Methods: Recombinant NNMT protein was used to prepare monoclonal antibodies by hybridoma technique. The diagnostic and prognostic function of NNMT protein in renal carcinoma was evaluated by analyzing 74 renal cancer tissues through immunohistochemical staining for NNMT by using the prepared antibodies. Results: Two hybridomas named 2F8 and 1E7 stably secreting the monoclonal antibodies were isolated successfully, and characters such as isotypes and specificity were determined. NNMT protein was significantly up-regulated in renal cancer and significantly associated with tumor histology and ages. The univariate survival analysis demonstrated that the pT-status, high levels of NNMT, and distant metastasis were significant prognosticators. Conclusion: NNMT is over-expressed in a large proportion in renal cell cancers. High NNMT expression is significantly associated with unfavorable prognosis. However, the prognostic value of NNMT needs further verification in larger sample sizes.     

破伤风类毒素单克隆抗体的制备及初步鉴定

目的：制备并鉴定破伤风类毒素单克隆抗体杂交瘤细胞株,为破伤风的快速检测及致病机制的研究提供实验材料。方法：用破伤风类毒素做抗原免疫BALB/C小鼠,取其脾细胞与小鼠骨髓瘤细胞融合,获得分泌高滴度针对破伤风类毒素的杂交瘤细胞株,测定单抗免疫球蛋白亚类及单抗效价,用间接ELISA和Western-blot检测单克隆细胞株的特异性。结果：通过细胞融合和克隆化,筛选出3株持续分泌抗破伤风类毒素单克隆抗体的杂交瘤细胞株1E12、1E11、1G10。间接ELISA和Western-blot检测结果表明1E12、1E11、1G10可以和破伤风类毒素发生特异性反应。结论：成功制备了抗破伤风类毒素单克隆抗体,为制备免疫诊断试剂盒和抗体药物的开发奠定了基础。     

P53基因蛋白在消化道肿瘤中的表达同浸润深度与转移的相关性观察

目的探讨P53基因表达与胃癌、肠癌、食管癌的相互关系.方法采用SP免疫组化染色方法.检测了消化道癌中(胃癌22例、肠癌22例、食管癌60例)P53蛋白的表达率.结果P53蛋白在胃癌中表达率最高达63.6%,肠癌中表达率为55.0%,食管癌中的表达率为61.7%.结论P53蛋白在消化道肿瘤中普遍表达并且当癌浸润较深和有淋巴结转移者阳性率较高.     

Advances in monoclonal antibody application in myocarditis

Monoclonal antibodies have become a part of daily preparation technologies in many laboratories. Attempts have been made to apply monoclonal antibodies to open a new train of thought for clinical treatments of autoimmune diseases, inflammatory diseases, cancer, and other immune-associated diseases. This paper is a prospective review to anticipate that monoclonal antibody application in the treatment of myocarditis, an inflammatory disease of the heart, could be a novel approach in the future. In order to better understand the current state of the art in monoclonal antibody techniques and advance applications in myocarditis, we, through a significant amount of literature research both domestic and abroad, developed a systematic elaboration of monoclonal antibodies, pathogenesis of myocarditis, and application of monoclonal antibodies in myocarditis. This paper presents review of the literature of some therapeutic aspects of monoclonal antibodies in myocarditis and dilated cardiomyopathy to demonstrate the advance of monoclonal antibody application in myocarditis and a strong anticipation that monoclonal antibody application may supply an effective therapeutic approach to relieve the severity of myocarditis in the future. Under conventional therapy, myocarditis is typically associated with congestive heart failure as a progressive outcome, indicating the need for alternative therapeutic strategies to improve long-term results. Reviewing some therapeutic aspects of monoclonal antibodies in myocarditis, we recently found that monoclonal antibodies with high purity and strong specificity can accurately act on target and achieve definite progress in the treatment of viral myocarditis in rat model and may meet the need above. However, several issues remain. The technology on how to make a higher homologous and weak immunogenic humanized or human source antibody and the treatment mechanism of monoclonal antibodies may provide solutions for these open issues. If we are to further stimulate progress in the area of clinical decision support, we must continue to develop and refine our understanding and use of monoclonal antibodies in myocarditis.     

胃癌患者血清中抗核抗体的检测

[目的]了解胃癌患者血清中抗核抗体谱的特点.[方法]以鸡卵核为抗原,应用免疫印迹法对18份胃癌术前患者以及22份正常对照组(年龄和性别分布均无差异)的血清进行分析.[结果]胃癌术前患者血清中特异识别5条抗原条带的抗核抗体的阳性率显著高于正常对照组,靶抗原的蛋白分子质量及胃癌术前组血清中特异识别这5条抗原条带的抗核抗体的阳性率分别为:69KD(44.4％,8/18),62KD(50％,9/18),58KD(33.9％,7/18),44KD(22.2％,4/18),37KD(38.9％,7/18).[结论] 胃癌患者体内存在抗核抗体,其相对的靶抗原有多种.     

MMP-9和CD15s与胃癌血管生成的相关性研究及其临床意义

目的:探讨基质金属蛋白酶9(MMP-9)和唾液酸化Lewis X(CD15s)抗原与胃癌侵袭转移的相关性。方法:采用CD105,MMP-9,CD15s进行免疫组化S-P法染色,对47例胃癌根治性切除手术标本病理组织切片进行标记,检测CD105的微血管密度(MVD),并对MVD与MMP-9、CD15s的关系进行统计分析。结果:MMP-9、CD15s表达都呈阳性的胃癌组织微血管密度高于MMP-9和CD15s单阳性及二者均阴性的胃癌组织。结论:MMP-9高表达有利于癌组织局部浸润,CD15s高表达有利于癌细胞血行转移,MMP-9及CD15s检测对临床诊治胃癌及判断预后具有一定意义。     

肾癌肿瘤浸润淋巴细胞免疫组化分析及其与肾癌愈后关系研究

目的:了解肾细胞癌(RCC)患者抗瘤免疫状况及TIL与肾癌愈后关系。方法:应用免疫组织化学PAP法和单克隆抗体对肾细胞癌原位肿瘤浸润琳巴细胞的构成与分布及活化进行研究,检测的TIL主要表面表标志包括CD_3、CD_4、CD_8、CD_(25)和HLA-DR抗原。结果:(1)RCC患者肿瘤原位TIL以·OT细胞为主(CD_3 ),其中CD_8 细胞明显高于CD_4 细胞,其CD_4 /CD_8 比值与RCC临床分期成负相关。(2)在肿瘤原位,有部分表达活化标志CD_(25)或HLA-DR抗原T细胞存在。(3)RCC患者其TIL在瘤组织中呈弥散状分布。结论 肾癌TIL反映了肾癌患者局部抗瘤免疫状况,且与分期负相关,可做为肾癌愈后的客观检测指标。     

肺和食道鳞状细胞癌中P27基因蛋白的表达

目的探讨肺和食道鳞状细胞癌中P27基因蛋白的表达状况.方法收集手术切除鳞癌标本63例,10%中性甲醛固定,石蜡切片,厚度5μm.超敏型S-P免疫组织化学染色.结果P27在鳞状细胞癌中总阳性率39.68%,在肺和食道中阳性率分别为36.00%和42.15%.在高、中分化组阳性率为36.00%,低分化组阳性率为53.85%.结论鳞癌组织中P27阳性表达与分化程度及是否转移的相关性有深入研究的必要.     

Monoclonal antibody-based ELISA and colloidal gold-based immunochromatographic assay for streptomycin residue detection in milk and swine urine

A protein conjugate of streptomycin (streptomycin-bovine serum albumin (BSA) conjugate) was prepared and used as immunogen to produce monoclonal antibodies (MAb). One hybridoma secreting anti-streptomycin MAb was obtained and then used to produce MAb. The MAb named 13H5 showed the 50% maximal inhibitory concentration (IC50) value of 4.65 ng/ml and the IC20 value of 0.21 ng/ml in phosphate buffered saline (PBS). At optimum conditions, an indirect competitive enzyme-linked immunosorbent assay (ELISA) and a co...     

子宫内膜癌PTEN蛋白表达及其临床意义

目的：探讨抑癌基因第10染色体同源丢失性磷酸酶一张力蛋白基因（PTEN）蛋白表达与子宫内膜癌的发生、发展的关系。方法：采用免疫组织化学S-P法检测PTEN蛋白在56例子宫内膜癌及30例正常子宫内膜组织的表达。结果：56例子宫内膜癌中PTEN蛋白表达缺失率为57．1％，明显高于正常子宫内膜中的完全表达（P〈0．05）。PTEN蛋白在低分化组的表达缺失率明显高于中高分化组（P〈0．05）。结论：抑癌基因PTEN蛋白表达缺失在子宫内膜癌的发生、发展、浸润等过程中有一定作用。     

p53基因蛋白在食道正常上皮及增生、癌变后表达的观察

目的 :探讨p5 3蛋白在食管癌组织发生中表达的意义。方法 :应用免疫组织化学技术检测 72例食管鳞状上皮癌 ,19例增生鳞状上皮及 17例正常食道鳞状上皮中 p5 3蛋白的表达。结果 :p5 3在食管癌组织中的阳性率为 4 7.95 % (35 / 72 ) ,在增生鳞状上皮中的阳性率为 36 .84 % (7/ 19) ,在正常食道鳞状上皮中的阳性率为 5 .88% (1/ 17) ,正常组与增生组和癌变组比较均有显著性差异 (P <0 .0 5 )。p5 3蛋白与癌的分化程度、淋巴结有无转移和浸润深度等指标均无明显关系。结论 :p5 3蛋白与食管癌的发生、发展有关。     

运用高通量测序研究Alport综合征多能干细胞的microRNA及其靶基因(英文)

目的:寻找来源于Alport综合征患者与正常对照者的多能干细胞差异性表达的microR NA,并对差异性表达的microR NA靶基因进行预测。创新点:本研究不同于一般的试验标本,试验标本是来源于尿肾脏管细胞诱导而成的多能干细胞。基于Alport综合征是遗传疾病,我们对一遗传家系进行了系统的分析。寻找特异性的差异性表达microR NA及其靶基因,从基因水平对Alport综合征进行研究。方法:在前期工作中,成功地从实验者与对照者的尿肾脏管细胞诱导成多能干细胞。运用高通量测序技术分析并发现实验者与对照者之间差异性表达的microR NA。对差异性表达的microR NA靶基因进行预测,并进行靶基因聚集分析,研究靶基因主要参与的生物学过程。同时进行靶基因信号传导通路的分析,研究靶基因主要参与的细胞信号传导通路。结论:在实验组与对照组之间,发现了30个具有显著差异性表达的microR NAs,包括19个上调表达与11个下调表达。差异性表达的microR NA的靶基因主要聚集在细胞膜和细胞代谢过程;靶基因主要参与嘌呤代谢通路与丝裂原活化蛋白激酶(MAPK)通路。     

单克隆抗体研究进展——不同表达系统介绍与比较

单克隆抗体的高度特异性使其在包括细胞生物学、基础医学、临床诊断等领域得到广泛应用,而各种表达系统的开发进一步促进了单克隆抗体及其衍生物的发展.对目前用于生产单克隆抗体的主要表达系统进行综述,同时比较各系统的特点和优缺点.     

Demonstration of a new biosensing concept for immunodiagnostic applications based on change in surface conductance of antibodies after biomolecular interactions

We report an important observation that the surface conductivity of antibody layer immobilized on polylysine-coated glass substrate decreases upon the formation of complex with their specific antigens. This change in conductivity has been observed for both monoclonal and polyclonal antibodies. The conductance of monoclonal mouse IgG immobilized on polylysine-coated glass substrate changed from 1.02×10⁻⁸ Ω⁻¹ to 1.41×10⁻¹¹ Ω⁻¹ at 10 V when complex is formed due to the specific biomolecular interactions with rabbit anti-mouse IgG F(ab′)₂. Similar behavior was observed when the same set up was tested in two clinical assays: (1) anti-Leishmania antigen polyclonal antibodies taken from Kala Azar positive patient serum interacting with Leishmania promastigote antigen, and (2) anti-p21 polyclonal antibodies interacting with p21 antigen. The proposed concept can represent a new immunodiagnostic technique and may have wide ranging applications in biosensors and nanobiotechnology too.     

上皮性卵巢癌中Skp2蛋白的表达及临床意义

目的：探讨Skp2蛋白表达与上皮性卵巢癌发生、发展及预后评价的意义。方法：采用免疫组化法检测15例卵巢上皮性良性肿瘤、10例卵巢上皮性交界性肿瘤及58例上皮性卵巢癌中Skp2蛋白的表达情况。结果：Skp2在卵巢上皮性良性及交界性肿瘤中的表达均为阴性，而在上皮性卵巢癌中高表达，阳性率为48．28％（28／58），Skp2在中低分化组、Ⅲ／Ⅳ期组及有淋巴结转移组分别高于高分化组（P〈0．01）、Ⅰ／Ⅱ期组（P〈0．01）及无淋巴结转移组（P〈0．01）。在不同年龄及组织类型的卵巢癌中表达无显著性差异。结论：Skp2在上皮性卵巢癌中表达，表达水平越高提示预后越差。     

慢性肾功能衰竭患者血浆脂蛋白(a)与纤溶活性的相关性

目的:研究慢性肾功能衰竭患者血浆脂蛋白(a)与纤溶活性的关系.方法:应用酶联免疫吸附法分别测定58例慢性肾功能衰竭患者及51名健康对照者的血浆脂蛋白(a)浓度,并以底物显色法测定其组织型纤溶酶原激活剂(tPA)及其抑制物(PAI)活性.结果:慢性肾功能衰竭患者血浆脂蛋白(a)浓度和PAI活性明显升高,tPA活性明显降低;脂蛋白(a)浓度与血浆尿素氮水平、PAI活性均呈正相关;脂蛋白(a)浓度与tPA活性呈负相关.结论:肾功能对血浆脂蛋白(a)浓度起一定的调节作用,脂蛋白(a)水平的升高与纤溶活性低下有关.     

Skp2在膀胱上皮癌中的表达及意义

目的：探讨细胞周期蛋白Skp2表达与膀胱移形细胞癌生物学行为的关系。方法：对73例膀胱移形细胞癌进行组织学分级及分期，另取22例正常膀胱组织作为对照。采用免疫组织化学方法S-P法检测各例组织中Skp2蛋白表达水平。并分析其表达水平与临床病理特征的关系。结果：Skp2蛋白阳性表达率为50．7％（37／73），Skp2蛋白表达与肿瘤组织学分级、病理分期显著相关。结论：Skp2是评估膀胱移形细胞癌恶性程度,估计预后有价值的指标。     

抗人双特异性磷酸酶DUSP18单克隆抗体的制备及鉴定

双特异性磷酸酶（Dual Specificity Phosphatase）是酪氨酸磷酸酶家族中的一员,这个家族中的成员既能对磷酸化酪氨酸去磷酸化,也能对磷酸化丝／苏氨酸去磷酸化．此外,它们还在有丝分裂原激活蛋白磷酸酶信号途径（MAPK）中起重要生理功能．为了制备抗人双特异性磷酸酶的单克隆抗体（mAb）,以DUSP18重组蛋白为抗原免疫BALB／e小鼠,通过B淋巴细胞杂交瘤技术制备抗相应磷酸酶的mAb．用Western blot检测mAb对重组蛋白和细胞中天然磷酸酶的反应性,共获得6株可稳定分泌抗磷酸酶mAb的杂交瘤细胞株,为进一步研究双特异性磷酸酶的去磷酸化作用机理以及其在有丝分裂原激活蛋白磷酸酶信号途径（MAPK）中的信号转导提供强有力的工具．