整合素受体β3亚单位在非酒精性脂肪性肝病肝组织中的表达

目的观察在非酒精性脂肪性肝病(NAFLD)不同疾病状态中整合素受体β3亚单位的表达情况。方法清洁级雄性新西兰白兔30只,随机分成两组,一组饲以正常饮食(n=6),另一组饲以高脂饮食(n=24)。高脂饮食组分别在饲养1、2、3个月时随机处死8只白兔,取肝组织行HE染色和天狼星红染色,根据病理结果计算NAFLD活动度积分(NAS)。Western印迹和实时PCR检测肝组织内整合素受体β3亚单位蛋白和mRNA表达,免疫组织化学检测β3亚单位在肝脏细胞内的表达情况。结果根据病理结果,高脂饮食白兔分成3组:单纯脂肪肝组(n=10)、无纤维化的NASt组(n=6)和伴有纤维化的NASH组(n=8),NAS积分分别为2.43±0.79,5.38±1.30和5.13±1.05。与正常对照组相比,单纯脂肪肝组整合素受体β3亚单位的蛋白和mRNA表达差异无统计学意义,而NASH-组的表达明显增加,并且随着肝纤维化的加重,表达量增加更显著(均P<0.05)。此外,与正常对照组相比,单纯脂肪肝组肝组织内表达整合素受体β3亚单位的细胞无明显增多;而NASH组阳性细胞显著增多(均P<0.05),其中伴有纤维化的NASH组阳性细胞最多;这些阳性细胞主要分布于肝细胞间隙内。结论与正常对照组和单纯脂肪肝组相比,NASH组肝组织内有着更显著的整合素受体β3亚单位的表达,并且表达量随着肝纤维化的出现而进一步增加。     

胃癌患者血清及癌组织中CD44v6表达及意义

[目的]检测胃癌患者血清及胃癌组织中CD44v6的表达及其与胃癌临床病理特征间的关系,进一步阐述CD44v6在胃癌发生发展中的作用。[方法]收集40例经病理证实的胃癌患者,20例正常健康体检者为对照组,20例取自距癌组织边缘5cm以上癌旁胃黏膜标本。采用双抗体夹心-酶联免疫吸附法检测40例胃癌患者术前和20例正常健康者血清中CD44v6水平,免疫组化SP法测定40例经手术切除的胃癌组织标本和20例癌旁胃黏膜标本中CD44v6的表达。[结果]①胃癌患者血清中CD44v6[(177.21±46.48)μg/L]高于正常健康者(P<0.01)。②癌组织中CD44v6的阳性表达率为77.5%,显著高于癌旁胃黏膜中的表达(P<0.01),CD44v6在血清和组织中的表达随组织学分级、临床分期的增高及有无淋巴结转移而升高,呈正相关性。[结论]CD44v6参与了胃癌的发生,并且可反映机体肿瘤的负荷状态及预测肿瘤的进展;检测CD44v6的血清含量可能为胃癌的诊断提供依据。     

Randomized phase II study of sunitinib versus standard of care for patients with previously treated advanced triple-negative breast cancer

PurposeThis randomized, open-label phase II study compared the efficacy of sunitinib monotherapy with that of single-agent standard-of-care (SOC) chemotherapy in patients with previously treated advanced triple-negative breast cancer (TNBC).MethodsPatients with advanced TNBC, relapsed after anthracycline- and taxane-based chemotherapy, were randomized to receive either sunitinib (37.5 mg/day) or the investigator's choice of SOC therapy. Progression-free survival was the primary endpoint.ResultsMedian progression-free survival was 2.0 months with sunitinib and 2.7 months with SOC chemotherapy (one-sided P = 0.888). Median overall survival was not prolonged with sunitinib (9.4 months) compared with SOC chemotherapy (10.5 months; one-sided P = 0.839). The objective response rate was 3% with sunitinib and 7% with SOC chemotherapy (one-sided P = 0.962).ConclusionsSunitinib monotherapy did not improve efficacy compared with SOC chemotherapy in patients with previously treated advanced TNBC, for which identification of effective treatments and therapeutic targets remains an urgent need.Trial registrationNCT00246571.     

Evaluation of the Early Access STR Kit v1 on the Ion Torrent PGM™ platform

The Early Access STR Kit v1 is designed to detect 25-plex loci with next generation sequencing (NGS) technology on the Ion Torrent PGM™ platform, including 16 of 20 expanded Combined DNA Index System (CODIS) core loci (CSF1PO, D1S1656, D2S1338, D2S441, D3S1358, D5S818, D7S820, D8S1179, D10S1248, D13S317, D16S539, D19S433, D21S11, TH01, TPOX and vWA), 8 non-CODIS core loci (D1S1677, D2S1776, D4S2408, D5S2500.AC008791, D6S1043, D6S474, D9S2157 and D14S1434) and Amelogenin. In this study, we compared the Early Access STR Kit v1 with the Ion Torrent™ HID STR 10-plex to find out its improvements and explored an appropriate analytical threshold to enhance the performance. In addition, seven experiments were conducted to evaluate the Early Access STR Kit v1 such as studies of repeatability, concordance, sensitivity, mixtures, degraded samples, case-type samples and pedigrees. Other than a little discordance (0.95%) with CE-STR results observed at D21S11, NGS-STR results correctly reflected the sample being tested. Repeatable results were obtained from both initial PCRs and emPCRs aside from a few variations of allele coverage. Full profiles could be obtained from 100 pg input DNA and >48.84% profiles from 10 pg input DNA. Mixtures were easily detected at 9:1 and 1:9 ratios. This system could be adapted to case-type samples and degraded samples. As a whole, the Early Access STR Kit v1 is a robust, reliable and reproducible assay for NGS-STR typing and a potential tool for human identification.     

High throughput ranking of recombinant avian scFv antibody fragments from crude lysates using the Biacore A100

Advances in molecular evolution strategies have made it possible to identify antibodies with exquisite specificities and also to fine-tune their biophysical properties for practically any specified application. Depending on the desired function, antibody/antigen interactions can be long-lived or short-lived and, therefore, particular attention is needed when seeking to identify antibodies with specific reaction-rate and affinity properties. Surface plasmon resonance (SPR) biosensors routinely generate sensitive and reliable kinetic data from antibody/antigen interactions for both therapeutic and diagnostic applications. However, many kinetic-based screening assays require rigorous sample preparation and purification prior to analysis. To ameliorate this problem, we developed a rapid and reliable assay for characterising recombinant scFv antibody fragments, directly from crude bacterial lysates. Ninety-six scFv antibodies derived from chickens immunised with C-reactive protein (CRP) were selected by phage display and evaluated using the Biacore A100 protein interaction array system. Antibodies were captured from crude bacterial extracts on the sensor chip surface and ranked based on the percentage of the complex left (% left) after dissociation in buffer. Kinetic rate constants (k(a) and k(d)) and affinity (K(D)) data were obtained for six clones that bound monomeric CRP across a broad affinity range (2.54 x 10(-8) to 3.53 x 10(-10) M). Using this assay format the A100 biosensor yielded high quality kinetic data, permitting the screening of nearly 400 antibody clones per day.     

Effects of Various Pharmaceutical Excipients on the Intestinal Transport and Absorption of Sulfasalazine,a Typical Substrate of Breast Cancer Resistance Protein Transporter

Breast cancer resistance protein (BCRP) transporter is an efflux transporter that utilizes energy from adenosine triphosphate hydrolysis to push its substrates, regardless of the concentration gradient. Its presence on the apical membrane of the intestinal mucosa is a major obstacle for the intestinal absorption of its substrates. In this study, we examined the effects of various pharmaceutical excipients on the intestinal transport and absorption of sulfasalazine, a BCRP substrate. Four excipients, including 0.05% and 0.075% BL-9EX, 0.01% and 0.05% Brij 97, 0.075% Labrasol, and 0.05% and 0.1% Tween 20 decreased the secretory transport of sulfasalazine in an in vitro diffusion chamber. Further investigation in an in situ closed loop experiment in rats showed that 0.05% and 0.1% BL-9EX and 0.1% Brij 97 effectively enhanced the intestinal absorption of sulfasalazine while maintaining minimal toxicity to the intestinal mucosa. However, 0.1% Brij 97 also increased the intestinal absorption of 5(6)-carboxyfluorescein, a paracellular marker compound. These findings suggest that BL-9EX might effectively inhibit the BCRP-mediated efflux of sulfasalazine in vivo, indicating that BL-9EX could improve the intestinal absorption of sulfasalazine and other BCRP substrates.     

The Burden of Peristomal Skin Complications on an Ostomy Population as Assessed by Health Utility and the Physical Component Summary of the SF-36v2®

Background

Body-altering surgery may affect perceptions of one’s self. For those with abdominal stoma surgeries, altered perceptions amplified by peristomal skin condition can increase health burdens.

毛重楼叶绿体基因组序列特征及其系统发育分析

目的获取毛重楼Parismairei叶绿体基因组信息特征并对其系统位置进行研究。方法采用Illumination测序技术对毛重楼叶绿体基因组进行测序,对其进行组装、注释和特征分析,并构建最大似然(maximumlikelihood,ML)系统发育树。结果毛重楼叶绿体基因组大小为163 918 bp,总GC含量为37.05%,大单拷贝区(large single-copy,LSC)、小单拷贝区(smallsingle-copyregion,SSC)分别为84173、13054bp,反向互补重复区(invertedrepeats,IR)大小为33296bp,共编码113个基因,包括79个蛋白质编码基因,30个t RNA基因和4个r RNA基因。在系统进化树中,重楼属Paris L.与藜芦科(Melanthiaceae Batsch ex Borkh.)关系较近,与百合科(Liliaceae Juss.)关系较远。结论 LSC区和SSC区序列变异高于IR区;相较于重楼属,毛重楼与蚤休组的亲缘关系最近,应将毛重楼归属为蚤休组;相较于百合科,毛重楼与藜芦科藜芦属亲缘关系更近,应将蚤休组从百合科中独立,归属为藜芦科。