基于TCMSP抗肿瘤中药小分子EGFR-TKI的研究

目的研究分析中药小分子表皮生长因子受体酪氨酸激酶抑制剂(EGFR-TKI)的抗肿瘤情况。方法挖掘中药系统药理学数据库与分析平台(TCMSP)中药小分子表皮生长因子受体抑制剂,利用反向分子对接服务器(DRAR-CPI)进行靶点验证。结果木犀草素、槲皮素、金雀异黄酮、表没食子儿茶素没食子酸酯、漆黄素5种中药小分子与表皮生长因子受体(EGFR)结合良好。结论中药小分子EGFR-TKI能从多个靶点、多种途径抑制肿瘤细胞的生长和扩散,利用中药小分子开发新一代多靶点EGFR-TKI具有广阔的前景。     

EGFR基因C端结构域真核表达载体的构建

构建EGFR基因C端结构域的真核表达载体.应用PCR技术,从含EGFR基因C端结构域的大肠杆菌DH 5α中扩增其序列,亚克隆到真核表达载体pcDNA3.1( )中,经酶切及测序进行验证.PCR扩增片段与预期结果相符,真核表达载体构建成功,测序结果与GenBank公布的基因一致.成功地构建了EGFR基因C端两个结构域的真核表达载体.     

Isolation of human epidermal stem cells by adherence and the reconstruction of skin equivalents

The isolation of human epidermal stem cells is critical for their clinical applications. In the present study, we isolated three populations of epidermal keratinocytes according to their ability to adhere to collagen type IV: i.e., rapidly adhering (RA), slowly adhering (SA), and non-adhering (NA) cells. The aim of this study was to characterize RA cells and to investigate the possibility of using these cells for epidermis reconstruction. To identify RA cells, flow cytometric analysis was performed using anti-₆ integrin and anti-CD71 antibodies. RA cells express high levels of ₆ integrin and low levels of CD71, which are considered as markers of an epidermal stem cell nature. Furthermore, electron microscopy showed that RA cells are small and have a high nuclear to cytoplasmic ratio, whereas SA and NA cells have well-developed cellular organelles and abundant tonofilaments. Western blot analysis showed that RA cells are slow cycling and express p63, a putative epidermal stem cell marker, whereas SA and NA cells express c-Myc, which is known to regulate stem cell fate. To compare epidermal regenerative abilities, skin equivalents (SEs) were made using RA, SA, and NA cells. The epidermis constructed from RA cells was well formed compared to those formed from SA or NA cells. In addition, only SEs with RA cells expressed ₆ integrin and ₁ integrin at the basal layer. These results indicate that RA cells represent epidermal stem cells and are predominately comprised of stem cells. Therefore, the isolation of RA cells using a simple technique offers a potential route to their clinical application, because they are easily isolated and provide a high yield of epidermal stem cells.Received 2 July 2004; received after revision 20 August 2004; accepted 10 September 2004     

人脐静脉内皮细胞原代培养方法的建立

目的：探索并建立人脐静脉内皮细胞（删Ⅶc）体外稳定培养的方法。方法：以1％Ⅱ型胶原酶消化收集人脐静脉内皮细胞，用嘲加10％的胎牛血清进行培养，免疫细胞化学方法鉴定内皮细胞。结果：原代培养的内皮细胞镜下呈典型的铺路石或者鹅卵石样排列，3～4d时生长最快。5d左右融合；免疫细胞化学染色显示内皮细胞Ⅷ因子相关抗原表达阳性。结论：酶消化法获得的原代血管内皮细胞有较高的存活率，可以作为基础和临床研究的良好模型。     

大袋蛾雌虫性信息素腺细胞的超微结构

<正>用透射电镜研究了大袋蛾性信息素腺细胞的超微结构。每个腺细胞具有一个表皮孔和一个由微绒毛围成的端器,但是没有导管细胞,也没有真正的表皮导管。腺细胞上方的表皮显然是由间细胞分泌形成的。腺细胞的分泌物从端器中经表皮孔而排到体外,所以这进一步证实了大袋蛾的性信息素腺细胞是一类新型的昆虫表皮腺细胞。基于各种昆虫表皮腺细胞的形态和功能,还讨论了昆虫表皮腺细胞的可能的进化途径。     

Effect of EGF1 peptides in directing nanoparticles to thrombi

In order to examine the effect of EGF1 peptides in directing nanoparticles to thrombi, the synthesized EGF1 peptide was conjugated to poly (ethyleneglycol)-poly (lactide) (PEG-PLA) nanoparticles (NP) to form EGF1-NP. A lipophilic fluorescent dye, coumarin-6, was incorporated into EGF1-NP so as to detect its loading and release capacity. The binding ability of EGF1-NP with TF-expressing cells was shown to be significantly higher than that of the non-conjugated NP. Following an intravenous administration, fluorescence was distributed along the vessel wall of the thrombosis regions in the model rats injected with coumarin-6-loaded EGF1-NP. The in vitro and in vivo results suggest that EGF1-NP is a promising drug delivery system for targeting cerebral thrombi.     

PIK3CA基因突变及PTEN表达与西妥昔单抗疗效的关系

 对西妥昔单抗治疗的KRAS野生型结直肠癌患者进行疗效评估及随访,收集每例患者的癌组织标本及临床病理资料,检测每例患者BRAF、PIK3CA基因的突变情况,采用免疫组织化学染色检测PTEN在癌及癌旁组织中的表达情况.结果显示,KRAS、BRAF及PIK3CA基因均为野生型的患者对单抗治疗的反应率为62.5%(CR+PR),而存在BRAF及PIK3CA突变患者的反应率分别为0,28.6%,证实BRAF及PIK3CA的突变可显著影响西妥昔单抗(C225)的疗效.PTEN主要表达在细胞核内,在癌组织中表达降低或缺失,在正常组织中表达正常,并且PTEN的表达与C225的疗效密切相关,PTEN表达阳性及阴性患者的反应率(CR+PR)分别为42.9%,36.4%(P＜0.05).BRAF、PIK3CA的突变和PTEN的表达缺失可以显著影响C225的疗效.     

根癌农杆菌介导的GFP在洋葱表皮细胞定位研究

采用根癌农杆菌介导的方法,以受控于CaMV35S启动子的携带有GFP报告基因的双元植物表达载体pCAMBIA1300-35S-GFP转化洋葱表皮细胞.荧光显微镜下观察结果显示,GFP基因在经浸染和共培养后的洋葱表皮细胞中得到了表达,绿色荧光分布在细胞核和细胞质中,为进一步研究新基因的亚细胞定位和瞬时表达奠定了基础.     

Localization of orphan receptor TR3 mRNA in early developmental follicles in rat

Byin situ hybridization, the localization of orphan receptor TR3 mRNA has been observed in early developmental follicles. TR3 mRNA is first expressed in the ovarian interstitial cells on day 2 after birth, and then in granulosa cells (GC) in primary follicles on day 4. The expression level of TR3 mRNA in GC increases following the follicular development. Its higher expression can be observed in the outer layer of GC and inner layer of theca cells (TC) on day 6, where the cells present active proliferation and differentiation. The expression of TR3 is in an increasing manner until the large antral follicles on day 30. The mRNA is only expressed in the healthy, but not atretic follicles in adult rat ovaries. Injection of epidermal growth factor (EGF) has dramatically enhanced its expression in the early stage of developmental follicles. It is therefore suggested that TR3 may play a role in regulating growth and differentiation of ovarian somatic cells in the early stage, and its expression is regulated by EGF.     

安徽堇菜属花粉与叶表皮微形态特征的研究

对安徽的堇菜属（Ｖｉｏｌａ）１６种花粉和２１种叶下表皮作了扫描电镜观察，花粉观察显示，花粉粒可分为两极均不沟通和具单极合流孔沟的三孔沟型两大类，根据它们的外壁纹饰，又可细分为皱块状纹型、致密穿孔型、链珠纹型、皱波纹型、网纹型、具孔穴的大皱块纹型及不规则小穿孔纹型７个类型。叶的表皮观察表明，下表皮角质层主要为条纹状加厚。作认为毛堇菜（Ｖ．ｃｏｎｆｕｓａ）是光瓣堇菜的（Ｖ．ｙｅｄｏｅｎｓｉｓ）异名，