基于直接和间接PPI信息预测人类核蛋白亚核定位

构建了含有五个亚核区域共426条蛋白质的单定位人类核蛋白数据库.选取蛋白质相互作用信息分数为信息参数,提出通过引入蛋白质一级间接相互作用预测蛋白质亚核定位的新算法,对人类核蛋白亚核定位进行了预测.对染色质、核仁两区域351条蛋白质的Jackknife检验总预测成功率为91.04%.对染色质、核仁、核基质、核斑四区域413条蛋白质的Jackknife检验总预测成功率为79.75%.

人类精子碱性核蛋白的毛细管电泳分析

目的 :建立毛细管电泳分离分析人类精子碱性核蛋白的新方法。方法 :Beckman聚丙烯酰胺涂层柱 ,总长 47cm ,有效长度 40cm。缓冲液 :2 0mmol·L-1柠檬酸 ,pH用 0 0 1mol·L-1氢氧化钠溶液调至 3 0 ;分离电压 :14kV ;柱温 :18℃ ;分离时间 2 0min ;检测波长 :2 14nm ;每次运行之间用缓冲液洗柱 4min ;压力进样 6～ 9s。结果 :所建立HPCE分析方法能够将HP1、HP2及HP3有效分离 ,迁移时间及峰高重现性分别小于 2 0 %及 10 % ,能够对大鼠及人精子碱性核蛋白进行快速分析。结论 :所建立方法比传统的凝胶电泳方便。

EBLN2基因对肝癌细胞增殖的抑制作用及其机制

目的探讨人类内源性非逆转录博纳病毒样核蛋白序列2(EBLN2)基因对肝癌细胞增殖的抑制作用及其机制。方法采用实时定量RT-PCR技术和蛋白印迹技术检测EBLN2基因在肝癌细胞系(Hep G2、SK-hep-1、Huh7、Hep3B、MHCC-97H、PLC/PRF/5、Focus、SMMC7721、YY-8103、LO2)和人肝癌组织样本中的表达;选取EBLN2表达水平较低的肝癌细胞株Focus和YY-8103进行外转质粒过表达,Pcmv-EBLN2重组质粒转入肝癌细胞株Focus和YY-8103,24 h观察EBLN2基因过表达对肝癌细胞株细胞增殖、平板克隆形成能力及软琼脂克隆形成能力、细胞周期的影响。结果定量和半定量PCR实验结果表明EBLN2基因在59%的肝癌样本中表达下调,且在10株肝癌细胞系(Hep G2、SK-hep-1、Huh7、Hep3B、MHCC-97H、PLC/PRF/5、Focus、SMMC7721、YY-8103、LO2)中表达水平与正常肝组织相比明显下调,依次下调48%、73%、72%、79%、84%、72%、87%、74%、82%、80%,P均<0.05;PcmvEBLN2质粒转染肝癌细胞株YY-8103和Focus 24 h后,肝癌细胞株YY-8103和Focus的细胞增殖率分别为72.2%、85.0%,平板克隆形成率分别为52.6%、25.6%,软琼脂克隆形成率分别为40.9%、37.5%,与空白对照相比,P均<0.05;肝癌细胞株YY-8103和Focus从G1期进入S期受到抑制。结论 EBLN2基因在肝癌组织中表达下调,其可能通过调节细胞周期抑制肝癌细胞增殖。

Lipid-associated membrane proteins of Mycoplasma penetrans induce production of proinflammatory cytokines in human monocytic cells

The aim of this study is to explore potential pathogenicity of Mycoplasma penetrans, and to investigate whether M. penetrans lipid-associated membrane proteins （LAMPs） could induce human monocytic cell line （THP- 1 ） to produce some proinflammatory cytokines in vitro, including interleukin- 1β （ IL- 1β）, tumor necrosis factor alpha （TNF-α）, and IL-8. THP-1 was stimulated with different concentrations of M.penetrans LAMPs and at different time to analyze the production of human IL-1β, TNF-α and IL-8. The protein levels of human IL-1β, TNF-α and IL-8 were measured by enzyme-linked immunoadsorbent assay （ELISA） and the mRNA levels of these proinflammatory cytokines were detected by reverse transcriptase-PCR （RT-PCR）. It was demonstrated in the present study that the production of IL-1β, TNF-α and IL-8 increased in dose- and time-dependent manner after stimulation with M. penetrans LAMPs in THP-1 cells. M. penetrans LAMPs also induced the expression of IL-1β, TNF-α and IL-8 mRNA. The production of IL-1β, TNF-α and IL-8 and the expression of mRNA were down-regulated by pyrrolidine dithiocarbamate （PDTC）. This study demonstrated that M. penetrans LAMPs can induce the production of proinflammatory cytokines in human monocytic cells in vitro, thus suggesting that it may be an important etiological factor.

Ribosome Inactivating Proteins from Plants Inhibiting Viruses

Many plants contain ribosome inactivating proteins (RIPs) with N-glycosidase activity, which depurinate large ribosomal RNA and arrest protein synthesis. RIPs so far tested inhibit replication of mRNA as well as DNA viruses and these proteins, isolated from plants, are found to be effective against a broad range of viruses such as human immunodeficiency virus (HIV), hepatitis B virus (HBV) and herpes simplex virus (HSV). Most of the research work related to RIPs has been focused on antiviral activity against HIV; however, the exact mechanism of antiviral activity is still not clear. The mechanism of antiviral activity was thought to follow inactivation of the host cell ribosome, leading to inhibition of viral protein translation and host cell death. Enzymatic activity of RIPs is not limited to depurination of the large rRNA, in addition they can depurinate viral DNA as well as RNA. Recently, Phase I/II clinical trials have demonstrated the potential use of RIPs for treating patients with HIV disease. The aim of this review is to focus on various RIPs from plants associated with anti-HIV activity.

STUDY ON NUCLEAR MATRIX PROTEINS FROM HUMAN BREAST CARCINOMA

Objective To investigate the marker protein of human breast carcinoma from nuclear matrix proteins (NMPs). Methods NMPs were injected subcutaneously into rabbit to get antiserum, which was used to detect the NMPs specificity for breast carcinoma. Results There was an apparent positive band (100 kD) in the NMPs of breast carcinoma, which did not exist in normal breast and other tumors that were detected.Conclusion One or one group of 100 kD NMPs were found to be related to human breast carcinoma, which may be involved in the carcinogenesis and development of human breast carcinoma and valuable for breast carcinoma diagnosis.