人表皮生长因子受体2抑制剂联合白细胞介素-2对人肾癌细胞株的疗效观察

目的：了解曲妥珠单抗（Trastuzumab，商品名为赫赛汀）联合白细胞介素-2（IL-2）对肾癌细胞在体外的杀伤抑制作用，同时检测肾癌细胞在药物作用前后人表皮生长因子受体2（HER2）、多药耐药相关蛋白1（MRP1）的表达情况。方法：肾癌细胞系通过细胞培养技术进行培养，用单四唑（MTT）法观察曲妥珠单抗联合IL-2对该细胞系的抑制杀伤效果，进而运用链霉抗生物素-过氧化物酶免疫组化（S-P）法对HER2、MRP1表达进行测定。结果：曲妥珠单抗联合IL-2对肾癌细胞的生长抑制及耐药性呈时间和剂量依赖性。经药物处理后，HER2、MRP1表达明显下降（P〈0．05）。结论：曲妥珠单抗能有效地抑制肾癌细胞的生长，对瘤细胞的耐药性也有一定的逆转作用。     

幽门螺杆菌cagA基因片段原核表达载体的构建及其cagA基因和CagA蛋白及感染者血清抗体的检测

目的:构建幽门螺杆菌(Helicobacter pylori,Hp)cagA基因片段的原核表达系统,建立ELISA检测CagA及其抗体,了解表达CagA的Hp菌株(CagA Hp)感染与所致疾病种类的关系.方法:从快速尿素酶试验阳性的156例患者胃黏膜活检标本中分离Hp.采用PCR检测109株Hp cagA基因,并从临床菌株Y06中扩增2 148 bp的cagA基因片段(cagA1).构建cagA1原核表达系统,用SDS-PAGE检查目的重组蛋白(rCagA1)的表达情况,用Western blot和免疫双扩散试验鉴定rCagA1的免疫反应性和抗原性.建立ELISA,检测109株Hp CagA表达和相应患者血清中CagA抗体.分析CagA Hp感染与胃炎和消化性溃疡的关系.结果:80.8%胃黏膜活检标本中分离出Hp(126/156),97.2%的Hp菌株(106/109)cagA基因阳性.与文献报道比较,所克隆的cagA1片段核苷酸和氨基酸序列同源性分别为94.83%和93.30%.所构建的原核表达系统表达的rCagA1量约为细菌总蛋白的30.0%.rCagA1能与Hp全菌抗体发生结合反应,免疫家兔产生双扩散效价为1∶4的抗体.92.6%菌株(101/109)可表达CagA,88.1% 患者(96/109)血清CagA抗体阳性.消化性溃疡标本中CagA Hp菌株分离阳性率(97.9%)高于胃炎标本(88.5%),但无统计学差异(χ2=3.48,P>0.05).结论:本实验构建的原核表达系统表达的rCagA1有良好的免疫反应性和抗原性,建立的ELISA可用于Hp CagA及其抗体的检测,分离的Hp菌株cagA基因携带率和CagA表达率均很高.消化性溃疡和胃炎与其感染的Hp是否表达CagA无明显关系.     

放疗诱导鼻咽癌细胞株CNE1热休克蛋白高表达

目的鼻咽癌是一种以放射治疗为主的头颈肿瘤,但部分鼻咽癌对放疗耐受。为了解鼻咽癌放疗耐受机制,本实验拟初步探讨人高分化鼻咽癌细胞株CNE1放疗后的蛋白质改变。方法利用固相pH梯度二维凝胶电泳,建立CNE1放疗前后总蛋白质2DE图谱,利用MALDI TOF MS及数据库搜索初步分析和鉴定部分放疗前后差异蛋白质点。结果高分子量酸性蛋白质在CNE1放疗后20min高表达,其中大多为热休克蛋白质HSPs(heatshockproteins,HSPs)。结论放射能诱导CNE1的热休克蛋白高表达,它们可能与鼻咽癌放疗耐受有关。     

Effects of histamine on growth and apoptosis of human melanoma cells A375

Histamine,as a biogenic amine,exists in bod-ies diffusely.Like other neural transmitters,his-tamine can play various physiological roles whencombining with special receptors on plasma mem-brane of target cells.Previous studies suggestedinconsistent results that histamine promoted or in-hibited growth of primary or metastatic melanomacells.The mechanism has not been elucidated ful-ly.The present research is aimed at investigatingwhether histamine has effects on growth and apop-tosis of human me…     

Anti-sense RNA Inhibits the Expression of Synaptotagmin Ⅱ in RBL-2H3 and Enhances the Exocytosis of Lysosomes in RBL-2H3

The expression of synaptotagmin Ⅱ (Syt2) in RBL-2H3 (RBL) and its role during exocytosis of RBL was investigated. The expression of Syt2 in RBL was detected by western blot and Syt2 gene was amplified by PCR. The anti-sense full length Syt2 cDNA expression vector was constructed with pEGFP-N1 and transfected into RBL by electroporation, and stable transfectants were selected by using G418. To analyze the role of Syt2 during exocytosis of RBL, the release of cathepsin D was assayed by immunoblotting. The results showed that Syt2 was expressed in RBL.The anti-sense expression vector pEGFP-N1-Syt2-AS was constructed and the sequence of insertion was completely consistent with rat Syt2 (accession number in GeneBank: NM012665). The stable transfectants (RBL-Syt2-AS) were obtained. Western blot showed that RBL-Syt2-AS expressed a lower level of Syt2 (8 % and 10 % of control cells), indicating that the expression of Syt2 in RBLSyt2-AS was markedly down-regulated by anti-RNA. Compared with control, the release of cathepsin D by RBL-Syt2-AS was increased. It was concluded that Syt2 expressed in RBL and could inhibit exocytosis of lysosomes in RBL.     

IL-2治疗对肝癌化疗敏感性影响的研究

目的　研究IL -2治疗与肝细胞癌化疗敏感性的关系。方法　采用S -P免疫组织化学方法对 14例术前有IL -2治疗史、12例术前没有IL -2治疗史的肝细胞癌存档石蜡包埋切片组织中MRP1、IL -2受体α链的表达进行检测。结果　IL -2治疗组中IL -2受体α链表达强度为 :0 3 0 19± 0 0 40 2 3 ,明显高于非治疗 (0 2 63 1± 0 0 2 2 2 4,P <0 0 1) ,IL -2治疗组IL -2治疗有效。IL-2治疗组中MRP1表达的强度为 :0 3 3 64± 0 0 44 67,非治疗组为 :0 3 0 0 8± 0 0 3 764 ,两者的差异有显著性意义 (P <0 0 5 )。结论　用IL -2治疗HCC患者 ,在增加机体抗肿瘤免疫的同时增加了肝癌细胞对化疗药物的耐受性 ,因此 ,在以化疗为主要治疗手段时 ,联合应用IL -2会降低治疗效果。     

亚砷酸对肝癌细胞增殖及PCNA,c-Myc蛋白表达的影响

目的:探讨亚砷酸对人肝癌BEL-7402细胞增殖及增殖细胞核抗原(PCNA),c-Myc蛋白表达的影响.方法:应用四氮唑盐(MTT)比色法、细胞群体倍增时间(TD)、细胞集落形成试验观察亚砷酸对BEL-7402细胞增殖的影响,免疫组织化学法观察亚砷酸对BEL-7402细胞PCNA,c-Myc蛋白表达的影响.结果:亚砷酸处理后BEL-7402细胞生长抑制率上升(2.0,4.0,8.0 μmol/L的亚砷酸作用24,48,72 h后抑制率分别为27.13%,42.65%,47.74%;35.52%,53.24%,60.41%;50.61%,65.88%,70.81%;与对照组比较P＜0.05),TD逐渐延长[2.0,4.0,8.0 μmol/L的亚砷酸作用48h后TD分别为(24.68±3.99);(32.42±4.46);(36.95±8.91);与对照组比较P＜0.05及P＜0.01],集落形成率下降(0,8.0 μmol/L的亚砷酸作用7d后集落形成率分别为46.00%,18.80%;P＜0.05),免疫组化结果显示亚砷酸处理后的BEL-7402细胞PCNA,c-Myc蛋白的表达明显减弱.结论:亚砷酸能抑制BEL-7402细胞增殖,其机制可能与PCNA,c-Myc蛋白的表达下调有关.     

大豆蛋白对胆固醇作用的研究进展

大量文献表明，大豆蛋白可以降低实验动物和人血液中胆固醇的含量，但其机理尚不完全清楚。为此，本文从胆固醇吸收、代谢、排泄及其对内分泌的影响等几方面详细地综述了膳食中大豆蛋白对胆固醇代谢的影响，并对其可能的作用机理加以阐述。     

补肾方对大鼠肝癌PCNA、p53蛋白的影响

目的:研究补肾方对二乙基亚硝胺(DEN)诱导大鼠肝癌过程中增殖细胞核抗原(PCNA)、(P53)蛋白表达的影响.方法:建立DEN诱导大鼠肝癌模型,分别于诱癌第12周末和第18周末采用免疫组化染色法检测补肾方治疗前后大鼠肝细胞PCNA、P53蛋白.结果:P53阳性细胞可见于癌前肝细胞的增生结节中;补肾方在诱癌第12周末和第18周末均能明显抑制P53的阳性表达(P<0.05),显著降低PCNA-LI(P<0.01).结论:在二乙基亚硝胺(DEN)诱导的大鼠肝癌模型中P53基因突变可发生于肝癌癌前病变,参与肝癌的启动过程.补肾方可抑制大鼠诱癌过程中肝细胞的增殖活性和P53的突变,从而延缓肝癌的形成.     

Immunohistochemical Study of Serum Proteins in Normal and Cadmium-Treated Mouse Testis by in vivo CryotechniqueImmunohistochemical Study of Serum Proteins in Normal and Cadmium-Treated Mouse Testis by in vivo Cryotechnique

Objective To investigate time-dependent changes of serum proteins permeability in the normal and cadmium（Cd）-treated mouse testis, reflecting tight junctional （TJ） barriers of Sertoli cells Methods The serum proteins, albumin and immunoglobulin-G（IgG）, in the seminiferous tubules were firstly immobilized by the “in vivo cryotechnique”, in which the dynamic blood circulation was always kept. The cryofixed testicular tissues were then processed for the freeze-substitution method, and embedded in the paraffin wax. Serial sections of 5μm thickness were immunostained by anti-mouse albumin or IgG antibody with peroxidase immunostaining, and also stained with hematoxylin-eosine （HE）for morphological observation. Results In normal seminiferous tubules, albumin immunoreaction products were localized around peritubular myoid cells and among Leydig cells, as well as in blood vessels. They were also localized as arch-like patterns around some spermatogonia in basal compartments. The number of the immunopositive arch structures was different according to developmental stages of the seminiferous cycle, judging from the arrangement of germ cells by HE-staining. The patterns of localization of IgG immunostaining in normal mouse testis were similar to that of albumin. In 24 h after Cd-treatment, some enlarged spaces and vesicular formation in the seminiferous epithelium were observed on the paraffin sections by HE-staining. The albumin or IgGimmunolocalization was seen not only in the basal compartments, but also in the adluminal compartments between Sertoli.cells and germ cells. Conclusion The structural changes of inter-Sertoli TJ barriers in vivo, such as different immunostaining patterns of serum proteins between the normal and Cd-treated mouse seminiferous tubules, could be clearly detected by the “in vivo cryotechnique” with albumin or IgG immunohistochemistry.