The effect of sulconazole on the antigenic composition of two dermatophyte species

Trichophyton rubrum and Trichophyton interdigitale have been grown in liquid culture in the presence of sulconazole. The antigenic activity of detergent extracts of intact organisms was analysed following SDS-PAGE and the probing of Western blots with homologous antisera raised in rabbits and with sera from patients with dermatophyte infections. Differences in protein-band patterns were noted; some bands present in control samples were absent in azole-treated samples and vice versa. These differences were reflected in antigenic band patterns, especially among components of approximate molecular weight of 30-40, 50-60 and 92-100 kDa.     

Serologische AIDS-Diagnostik mit gentechnisch gewonnenen Polypeptiden des menschlichen Immundefizienz-Virus (HIV-1)

Summary Serologic testing for human immunodeficiency virus type 1 (HIV-1) is currently based on enzyme linked immunosorbent assay (ELISA) as screening method. Positive ELISA-results have to be confirmed by at least one second procedure such as Western blotting or immunofluorescence. To obtain new diagnostic reagents for confirmatory testing, we expressed viral antigens in procaryotic systems. Peptides representing epitopes of structural core (gag)- and envelope (env)-proteins of HIV were produced inE. coli as stable immunogenic -galactosidase fusion proteins. Recombinant proteins were taken for immunoblot-assays. The results of Western blotting with those fusion proteins were in general comparable with conventional ELISA, immunofluorescence, immunoblot with cell-culture derived virus and commercially available ELISA tests based on recombinant proteins. Immunoblots using recombinant transmembrane protein (gp41) derived polypeptide were more sensitive than the conventional procedure with purified virion proteins. Western blotting with recombinant fusionproteins provide reliable and inexpensive serodiagnostics without handling of infectious cell cultures.

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Abkürzungsverzeichnis core- (gag-) Proteine Innere Virus-Strukturproteine - HIV-I Humanes Immundefizienz-Virus Typ 1 - ELISA Enzyme linked immunosorbent assay - enve- lope- (env-) Proteine Äußere Virus-Hüllproteine - gp 41 Glykoprotein 41 - p 24 Protein 24     

Das erregerspezifische Antikörperprofil in den verschiedenen Stadien der HIV-1-Infektion

Summary The westernblot analysis of 170 patients with HIV-1-infection demonstrated that 47% of the patients in latent stage, 58% of the patients with lymphadenopathy-syndrome and only 25% of the patients with the full-blown picture of AIDS showed the complete pattern of HIV-specific antibody response. This antibody response is mainly directed against the env-encoded envelope proteins gp160, gp120 and gp41, against the gag-encoded core proteins p55, p24 and p17 as well as against the pol-encoded enzymatic proteins p66, p51 and p31.Antibodies against gp160 and gp120 were present in nearly all patients, whereas the prevalence of the other antibodies decreased with the stage of the disease. Statistical significant differences were found particularly between patients with LAS or AIDS respectively. Antibodies against p17 were detected in 74% of the patients with LAS but only in 25% of the patients with AIDS. The lack of antibodies against p17, p24 or p51 was significantly associated with lower mean CD4/CD8-ratios (p<0,007) and higher mean serum levels of IgA (p<0,001) and beta-2-microglobulin (p<0.001). One third of the patients with LAS and this reduced pattern of antibody response developed AIDS within six months. These results demonstrate that the detection of antibodies against p17, p24 or p51 is of prognostic importance. A serological profile which lacks the antibody response against at least two of those three viral antigens indicates a progression of the disease activity.

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Abkürzungsverzeichnis LAS Lymphadenopathie-Syndrom - ELISA Enzyme-linked-Immunosorbent-Assay Herrn Prof. Dr. N. Zöllner zum 65. Geburtstag gewidmet     

HSP90β反义核酸载体转染细胞HSP90β蛋白的表达

目的了解HSP90β 反义核酸转染细胞中HSP90β蛋白的表达。方法通过lipofectamine介导将HSP90β反义核酸重组子pcDNA HSP90转染人胃癌细胞系SGC7901、人胃癌多药耐药细胞系SGC7901/VCR、人肝癌细胞系HCC7402及人食管癌细胞系Ec109。经用G418进行筛选 ,对筛选出的阳性克隆用Westernblot检测HSP90β蛋白的表达。结果pcDNA HSP90转染人胃癌细胞系SGC7901 ,人胃癌耐药细胞系SGC7901/VCR ,人肝癌细胞系HCC7402及人食管癌细胞系Ec109后 ,用G418筛选出的阳性克隆 ,分别被命名为:AH SGC7901 ,AH SGC7901/VCR ,AH HCC7402及AH Ec109。Westernblot检测结果表明 ,AH SGC7901,AH SGC7901/VCR ,AH HCC7402及AH Ec109表达的HSP90β蛋白低于其亲本细胞。结论HSP90β反义核酸可封闭HSP90βmRNA ,使pcDNA HSP90转染的细胞AH SGC7901 ,AH SGC7901/VCR ,AH HCC7402及AH Ec109表达的HSP90β蛋白减少 ,为研究HSP90下调对肿瘤细胞生物学活性的影响提供了实验材料。     

全反式维甲酸对白血病NB4细胞PRAM蛋白的影响

目的:探讨在NB4细胞增殖过程中PRAM蛋白表达情况,并且用全反式维甲酸（ATRA）进行干扰,观察其对PRAM蛋白的影响。方法:①细胞增殖抑制实验:体外培养NB4细胞共5天,用台盼蓝染色法筛选ATRA有效干预浓度后,选取ATRA(1×10^-6mol/L)为目的干预浓度。②Western blot:ATRA(1×10^-6mol/L)持续干扰NB4细胞1、2、3天后检测PRAM蛋白的相对表达量。结果:①与对照组比较,4种浓度ATRA组细胞与NB4细胞的增殖率呈量效与时效关系;浓度越大,抑制率越高;于培养第4天出现最强抑制。②选取1×10^-6mol/L ATRA干扰NB4细胞共3天,对照组及ATRA组PRAM蛋白的表达在时间上有规律性:第2天表达最强烈。③1×10^-6mol/L ATRA使PRAM蛋白在第2天的相对表达量较对照组低（P<0.05）。④1×10^-6mol/l ATRA使NB4细胞PRAM蛋白在第2天出现最大表达量,同时该组细胞出现生长抑制。结论:ATRA能通过PRAM位点来达到抑制APL增殖分化的目的。     

苦参总碱体外抗柯萨奇B病毒3型作用测定及其机理的初步研究

先用酸水浸泡、氯仿萃取等处理提取苦参总碱，然后通过微量细胞培养法，病毒蛋白质Ｗｅｓｔｅｒｎｂｌｏｔ，分析了不同浓度苦参总碱对病毒增殖和衣壳蛋白含量的影响，表明苦参总碱在体外试验中能抑制柯萨奇Ｂ病毒３型（ＣＶＢ３）的增殖，并呈一定的剂量依赖性。通过病毒吸附和穿入细胞前后苦参总碱作用的比较，显示苦参总碱能进入细胞内发挥抗病毒作用，它可能不影响ＣＶＢ３吸附、穿入等环节，而是影响ＣＶＢ３侵入细胞后某环节，特别是病毒的生物合成。     

大蒜辣素致肝癌细胞HepG2凋亡研究

目的 研究大蒜辣素(Allicin)导致肝癌细胞HepG2凋亡的现象及其影响规律.方法 运用CCK-8试剂盒研究Al-licin对HepG2细胞增殖的影响,运用流式细胞仪检测其对细胞凋亡、DNA代谢的影响,运用免疫印迹(Western blot)研究肝癌细胞凋亡基因表达的影响.结果 Allicin能抑制HepG2细胞的增殖,细胞倍增时间延长为4 d,同时Allicin也能导致细胞出现大量凋亡,实验结果表明其凋亡率最高为4.13%,死亡率达到70.38%.细胞周期实验结果表明Allicin能使大部分细胞处于DNA合成期的S期及G0/G1期,延缓细胞增殖.Western blot实验结果表明Allicin能激活细胞凋亡基因提高药物的治疗效果.结论 Allicin能抑制肝癌细胞的增殖可导致细胞大量死亡并引起细胞凋亡,并能引起细胞DNA代谢发生紊乱,细胞大部分被阻滞在DNA合成的G0/G1期,细胞分裂被抑制,Western blot实验结果表明细胞凋亡基因大量表达,细胞发生凋亡.     

焦炉工血清p53蛋白的检测分析

目的 探讨职业接触焦炉逸散物的焦炉工人血清p53蛋白的表达水平及其在肺癌高危人群筛检中的意义.方法 运用蛋白质印迹技术(Westernblot),对接触组118例和对照组50例人群的血清p53蛋白进行检测,比较两组血清p53蛋白的表达水平.结果 接触组不同工龄组、不同作业地点间p53蛋白的表达水平具有统计学意义(P﹤0.05).结论 经常大量接触焦炉逸散物可使p53蛋白的表达水平升高,增加p53基因异常的危险性,从而使焦炉工患肺癌的危险性增加.     

Notch信号通路在培养神经元细胞中的表达

目的 观察在体外培养的神经元细胞内Notch基因的分布和表达.方法 原代神经元培养,免疫组织化学的方法定位.对按不同时间点进行划伤处理的神经元细胞进行Westernblot分析,测定其Notch表达的变化趋势.结果 在培养的神经元细胞内有大量细胞(＞99％)均为Notch基因表达信号阳性;按时间点损伤的培养细胞其Notch 1受体胞内段(N1CD)表达呈动态变化.结论 成熟神经元中存在Notch基因的表达,Notch基因可能参与神经元损伤的病理过程.