Targeting efficiency of RGD-modified nanocarriers with different ligand intervals in response to integrin αvβ3 clustering

Receptor change induced by ligand binding is a new issue to face in the field of targeted delivery. Receptor clustering, the main pattern of receptor changes, decreases the affinity between ligand and receptor due to the redistribution of receptor position. In an attempt to respond to such challenge, we designed and constructed three RGD-modified nanocarriers with different ligand intervals: stealth liposomes modified with the monomeric RGD (moRGD-LP), dimeric RGD (diRGD-LP) and a special dimeric RGD with a linker between two cyclic RGD motifs (P-diRGD-LP). The αvβ3-positive and -negative tumor cells (Melanoma B16 and MCF-7) were used as the cell models. As a result, P-diRGD-LP demonstrated strongest interaction with B16 cells in surface plasmon resonance study and highest cellular uptake in B16 cells in real-time confocal analysis. The enhanced endocytosis of P-diRGD-LP was found to be αvβ3-mediated and P-diRGD-LP increased the involvement of the clathrin-dependent pathway. Importantly, P-diRGD-LP demonstrated the best targeting effect in B16-tumor bearing mice in both in vivo and ex vivo near-infrared fluorescent images, about 2.4-fold that of moRGD-LP and 2.8-fold that of diRGD-LP at 3 h. Further, we validated integrin αvβ3 clustering on B16 cells via a single-molecule imaging by a total internal reflection fluorescence microscopy. Finally, the 3D models of αvβ3 clustering suggested a receptor interval within 41.916–65.779 Å, while the molecular computation revealed an RGD ligand interval of 20.944 Å, 42.753 Å and 78.196 Å for diRGD-LP, P-diRGD-LP and moRGD-LP, respectively, confirming the best matching between clustered αvβ3 and P-diRGD-LP. In conclusion, P-diRGD-LP could achieve higher targeting to αvβ3-positive tumor via the enhanced interaction based on the better ligand-receptor compatibility. The design of targeted nanocarriers against receptor clustering might provide new insight into the nanotechnology-based anticancer therapy.     

CD44v6和E-cadherin表达与结直肠癌浸润转移关系

目的　探讨CD44v6和E cadherin(E cad)蛋白表达与结直肠癌浸润转移的相关性。方法　应用免疫组织化学技术 ,检测 90例结直肠癌组织中CD44v6和E cad蛋白表达。结果　 90例结直肠癌中CD44v6和E cad蛋白阳性表达率分别为75 6 %和 46 7%。CD44v6高表达及E cad低表达与结直肠癌Dukes分期、浆膜浸润、淋巴结转移、肝脏转移均呈正相关 (P <0 0 5 )。结直肠癌中CD44v6表达与E cad表达呈负相关 (r =- 0 4 3,P <0 0 0 5 )。结论　CD44v6和E cad表达与结直肠癌浸润转移密切相关。检测CD44v6和E cad蛋白表达可作为判断结直肠癌预后的客观指标。     

鼠源轮状病毒vp7基因的表达载体构建及转基因植物的研究

目的：构建表达鼠源轮状病毒抗原基因vp7的植物表达载体及植物转化研究。方法：抗原基因vp7经PCR扩增后直接克隆到PUC－T载体，双酶切目的片断和植物表达载体，以T4连接酶将目的片断与载体相连接，电转化方式将重组质粒转入农杆菌。结果：以重组农杆菌为介导将靶基因转入到马铃薯外植体。成功地将目的基因定向克隆到表达载体，并转入到农杆菌中；以农杆菌为介导获得抗性转基因马铃薯植株。结论：通过PCR检测，初步确定轮状病毒外壳蛋白基因vp7已成功地整合到马铃薯植株中。     

Characterization of the hog cholera virus 5′ terminus

Hog cholera virus (HoCV) 5 terminus of the ALD and GPE(–) strains were analyzed by using rapid amplification of cDNA end method (5RACE). An additional nine nucleotides were found at the 5 termini of genomic RNA in the ALD and GPE(–) strains of HoCV. These nine nucleotides were also conserved in BVDV and were suggested to form a hairpin structure at the 5 terminus by computer-assisted analysis. It seems possible that the secondary structure and/or the 5 terminus sequence has a significant role in the HoCV virus genome.     

北京东灵山与雾灵山地区不同生境下柴胡中柴胡皂苷的差异

目的:探讨北京东灵山与雾灵山地区不同生境柴胡化学成分的差异,为认识和发展道地柴胡提供理论依据.方法:对东灵山和雾灵山不同生境类型的柴胡居群进行取样,采用HPLC分析不同生境下柴胡皂苷a和柴胡皂苷d含量的差异.结果:不同生境下柴胡皂苷含量差异显著,草甸生境下的柴胡皂苷含量较高,而林下生境及灌丛生境柴胡皂苷含量相对较低,经分析柴胡皂苷含量与坡向呈正相关,通风透光的生境条件有利于柴胡皂苷的累积.结论:生境对柴胡皂苷产生与积累起着非常重要的作用,建议考察道地药材,应突破传统区域认识,结合生境层级开展.     

宫颈癌组织中survivin和CD44v6的表达及其与HPV16/18感染的相关性

目的研究survivin、CD44v6在宫颈癌组织中的表达及其与HPV16/18感染的相关性,以探讨宫颈癌的发生机制。方法用免疫组化方法进行survivin和CD44v6的检测,用PCR检测HPV16/18感染情况。结果survivin和CD44v6的阳性率在宫颈癌组织中远高于CIN和正常宫颈组织,差异显著(均P<0.01),其表达率与淋巴结转移有关(均P<0.05)。CD44v6随着临床分期、病例分级的升高阳性率升高(P<0.05),survivin的阳性率则随着病例分级的升高而增加(P<0.05)。HPV16/18在正常宫颈组织、CIN和宫颈癌中的阳性率逐渐升高(P<0.01),但与临床分期、病理分级、淋巴转移无关。survivin与HPV16/18感染有相关性(P<0.05)。结论宫颈癌组织中survivin的异常表达与HPV16/18感染有关。survivin和CD44v6与宫颈癌的恶变程度有关,可作为宫颈癌筛查预后判断的有利指标。     

2017-01目录

目的 研究99Tcm-联肼尼克酰胺-3聚乙二醇-精氨酸-甘氨酸-天冬氨酸环肽二聚体（99Tcm-3PRGD2）整合素受体显像在乳腺癌定性诊断中的价值及与钼靶检查的对比。方法 选取初诊乳腺占位患者84例,均为女性。所有患者行2 h、4 h 99Tcm-3PRGD2 整合素受体显像及钼靶检查,以术后病理结果作为“金标准”,比较不同显像方法（全身显像及胸部SPECT/CT显像）对乳腺癌定性诊断的价值,并与钼靶结果进行对比分析。采用SPSS22.0软件,对数据进行t检验和χ2检验。结果 SPECT/CT显像的诊断效能高于全身显像;乳腺癌99Tcm-3PRGD2摄取率[病灶容积最大计数（T_max）、病灶容积平均计数（T_mean）、T_max/本底平均计数（B）值及T_mean/B值]均明显高于良性占位（t=2.09~3.19,均P＜0.05）,且T_max/B值差异更为明显（t=3.19,P＜0.01）;2 h、4 h显像对乳腺癌诊断效能差异无统计学意义（t=0.63~1.25,均P>0.05）。SPECT/CT显像诊断乳腺癌的灵敏度大于95.0%,联用钼靶后灵敏度可以提高到98.4%;对致密型乳腺患者,SPECT/CT显像与钼靶诊断乳腺癌的灵敏度、特异度、准确率分别为95.2%、83.3%、91.7%和78.6%、76.2%、77.8%,前者的准确率高于后者,且差异有统计学意义（χ2=4.341,P＜0.05）。结论 99Tcm-3PRGD2 SPECT/CT显像定性诊断乳腺癌具有高灵敏度,是对钼靶的有益补充,尤其对致密型乳腺占位患者具有独特优势。     

Animal models to test drugs with potential antidiabetic activity

Although medicinal plants have been historically used for diabetes treatment throughout the world, few of them have been validated by scientific criteria. Recently, a large diversity of animal models has been developed to better understand the pathogenesis of diabetes mellitus and new drugs have been introduced in the market to treat this disease. The aim of this work was to review the available animal models of diabetes and some in vitro models which have been used as tools to investigate the mechanism of action of drugs with potential antidiabetic properties. In addition, a MEDLINE/PUBMED search for articles on natural products, pancreatectomy and diabetes mellitus treatment published between 1996 and 2006 was done. In the majority of the studies, natural products mainly derived from plants have been tested in diabetes models induced by chemical agents. This review contributes to the researcher in the ethnopharmacology field to designs new strategies for the development of novel drugs to treat this serious condition that constitutes a global public health.     

联合检测胃癌患者外周血sCD44v5和CD44v5含量的意义

目的联合检测胃癌患者血清中可溶性CD44变异体5(sCD44v5)含量及外周血淋巴细胞(PBL)CD44变异体5(CD44v5)含量,探讨其意义。方法采用酶联免疫吸附测定(ELISA)法检测40例胃癌患者术前血清中sCD44v5含量。同时应用流式细胞仪(FCM)检测36例胃癌患者术前PBLCD44v5含量并与健康组相对照。结果胃癌血清中sCD44v5含量(平均36.308ng/ml)明显高于正常对照组(平均30.946ng/ml)(P<0.05)。无转移组sCD44v5含量(平均30.981ng/ml)与正常对照组相比,无显著差异(P>0.05)。但是,胃周转移组sCD44v5含量(平均39.811ng/ml)明显高于无转移组(P<0.01)以及远处转移组sCD44v5含量(平均27.733ng/ml,P<0.05)。胃癌患者术前PBLCD44v5含量[(7.138±5.461)%]较正常对照组[(2.741±1.842)%]明显增高(P<0.01)。有转移患者PBLCD44v5含量[(8.130±5.834)%]明显高于无转移者[(4.167±2.159)%](P<0.01)。结论胃癌患者血清中sCD44v5含量及术前PBLCD44v5含量似乎与转移程度关系密切。联合检测其含量可望为判断胃癌患者有无转移、转移程度、制定个体化治疗方案提供参考指标。     

重组质粒pBud-CD44v6的构建及其在胃癌细胞内的表达

目的 构建含人的肿瘤侵袭和转移的特异性抗原CD44v6编码基因的真核表达的重组载体,进行核苷酸序列分析,并在肿瘤细胞中表达.方法 ①以pBud作为真核表达载体,选择CD44v6编码基因进行RT-PCR扩增,构建重组表达质粒,进行酶切鉴定和测序分析.②在Lipofectamine 2000的介导下,将构建成功的pBud-CD44v6重组表达质粒转染到肿瘤细胞中,通过免疫组织化学方法检测CD44v6的表达.结果 经酶切鉴定和测序分析表明,插入的目的 基因片段为CD44v6的编码基因,长为129 bp,与GenBank中登录的cDNA相比较,同源性高达100%,且方向正确;重组质粒pBud-CD44v6转染对数生长期胃癌细胞株SGC-7901,进行免疫组织化学检测结果显示,与空质粒转染组对比,重组质粒组肿瘤细胞的细胞质中可见大量CD44v6基因编码的表达,其蛋白呈棕黄色,而对照组呈阴性.结论 成功构建了pBud-CD44v6真核表达的重组载体,并在肿瘤细胞中表达.