Intrinsic caspase-8 activation mediates sensitization of erlotinib-resistant tumor cells to erlotinib/cell-cycle inhibitors combination treatment

Inhibitors of the tyrosine kinase activity of epidermal growth factor receptor, as erlotinib, have an established role in treating several cancer types. However, resistance to erlotinib, particularly in breast cancer cell lines, and erlotinib treatment-associated disorders have also been described. Also, methods and combination therapies that could reverse resistance and ameliorate non-desirable effects represent a clinical challenge. Here, we show that the ATP non-competitive CDK2/cyclin A inhibitor NBI1 sensitizes erlotinib-resistant tumor cells to the combination treatment (co-treatment) for apoptosis-mediated cell death. Furthermore, in erlotinib-sensitive cells, the effective dose of erlotinib was lower in the presence of NBI1. The analysis in the breast cancer MDA-MB-468 erlotinib-resistant and in lung cancer A549 cell lines of the molecular mechanism underlying the apoptosis induced by co-treatment highlighted that the accumulation of DNA defects and depletion of cIAP and XIAP activates the ripoptosome that ultimately activates caspases-8 and -10 and apoptosis. This finding could have significant implications for future treatment strategies in clinical settings.     

Simple and rapid preparation of gapped plasmid DNA for incorporation of oligomers containing specific DNA lesions

The cytotoxicity, mutagenicity, and carcinogenicity of DNA base lesions are largely determined by the responses of cellular DNA repair proteins, DNA polymerases, and signaling pathways. Elucidation of these processes is thus of high biochemical interest. Such studies increasingly rely on DNA substrates containing specific lesions at defined locations. Although short synthetic DNA oligomers have frequently proved useful, circular plasmid substrates are preferable for much biochemical work, and essential for in vivo studies. However, the complexity of current approaches for preparing such substrates and limitations inherent in the procedures have posed problems. We present here a simple, highly versatile procedure for preparing gapped duplex plasmids, into which oligomers incorporating specific lesions can easily be inserted. Endonuclease N.BstNBI was used to nick twice the same strand of a pUC19-derived plasmid (pUC19HBDa), at two GAGTCNNNN sequences separated by 22 bases. Removal of the 22-nt oligomer and further purification produced a highly pure gapped plasmid. To illustrate application of this procedure, 22-nt oligonucleotides containing a single uracil residue were ligated into the gapped molecules. The pUC19HB(Da) plasmid can be modified to accept almost any DNA-lesion-containing oligomer. Using this new approach to incorporate specific DNA lesions into popular reporter genes will facilitate in vivo study of cellular responses to DNA damage.     

胃镜对咽喉部恶性病变诊断的临床应用价值

目的:探讨胃镜在对于咽喉部恶性疾病诊断的临床应用价值。方法:对2013年11月至2014年9月因声音嘶哑或咽部异物感或吞咽困难为主诉就诊的患者,给予电子喉镜或间接喉镜检查,以在喉镜下发现咽喉部肿物或喉镜下发现黏膜异常增生不能明确判定病灶性质的12例患者为研究对象。继而给予窄带成像(Narrow-band imaging,NBI)及放大胃镜检查,对病灶进行性质及病变范围的判定。病理结果作为金标准,比较喉镜及胃镜两种方式对咽喉恶性病灶的正确诊断率。结果:12例患者中共发现15处病灶,其中炎性反应5处,单纯性鳞状上皮增生3处,鳞状细胞癌6处,神经纤维瘤1处。喉镜对病灶的正确诊断率是40.0%(6/15),NBI放大胃镜对病灶的正确诊断率是93.3%(14/15),两者差异有统计学意义(x~2=9.60,P=0.005)。结论:NBI放大胃镜对咽喉部肿瘤诊断正确率高于普通喉镜,充分证明了胃镜在咽喉部恶性疾病的诊断方面的临床价值。在行胃镜检查时,同时对咽喉部进行仔细观察检查是十分必要的。     

Corticotropin-releasing factor, vasopressin and receptor systems in depression and anxiety

Summary. Affective disorders tend to be chronic and life-threatening diseases: suicide is estimated to be the cause of death in 10–15% of individuals with major depressive disorders. Major depression is one of the most prevalent and costly brain diseases with up to 20% of the worldwide population suffering from moderate to severe forms of the disease. Only 50% of individuals with depression show full remission in response to currently available antidepressant drug therapies which are based on serendipitous discoveries made in the 1950s. Previously underestimated, other severe depression-associated deleterious health-related effects have increasingly been recognized. Epidemiological studies have provided substantial evidence that patients with depression have a 2–4-fold increased risk both of developing cardiovascular disease and of mortality after experiencing a myocardial infarction. The majority of patients suffering from affective disorders have measurable shifts in their stress hormone regulation as reflected by elevated secretion of central and peripheral stress hormones or by altered hormonal responses to neuroendocrine challenge tests. In recent years, these alterations have increasingly been translated into testable hypotheses addressing the pathogenesis of illness. Refined molecular technologies and the creation of genetically engineered mice have allowed to specifically target individual genes involved in regulation of corticotropin releasing factor (CRF) and vasopressin (AVP) system elements. The cumulative evidence makes a strong case implicating dysfunction of these systems in the etiology and pathogenesis of depression and pathological anxiety. Translation of these advances into novel therapeutic strategies has already been started.     

色素内镜、窄带成像内镜对大肠平坦型病变检出的对比性研究

目的:探讨色素内镜(CE)、窄带成像内镜(NBI)对大肠平坦型病变的检出及相关因素。方法:对2010年4月至6月来我院国际医学中心的276例患者行结肠镜检查,262例患者入选。先行常规白光模式插镜至回盲部,应用NBI模式退镜观察,检出平坦型病变后记录病变特征(包括数量、部位、形态、大小),然后再在常规白光模式下插镜至回盲部,应用0.25%靛胭脂染色后行色素内镜退镜观察,检出平坦型病变后记录病变特征,最后于病变活检。比较CE和NBI对平坦型病变的检出并分析相关因素。结果:262例患者中,共计检出病变198个,其中平坦型病变69个,检出率为34.8%。NBI检出平坦型病变51个(73.9%);CE检出平坦型病变69个(100%)(P<0.05)。NBI漏诊了18个平坦型病变,漏诊率为26.1%。NBI检出的平坦型病变平均大小为8.7±2.6mm,CE检出的平坦型病变平均大小为4.5±2.1mm(P<0.05)。NBI检出<5mm的平坦型病变5个(7.2%),CE检出<5mm的平坦型病变19个(27.5%)(P<0.05)。NBI检出的IIc型平坦型病变13个(18.8%),CE检出的IIc型平坦型病变25个(36...     

CRF R1受体拮抗剂对幼年大鼠快眼动睡眠的影响

目的：观察腹腔注射CRFR1受体拮抗剂NBI27914对幼年大鼠睡眠/觉醒周期的影响。方法：生后13d的大鼠行电极安装术,术后12h开始多导睡眠描记,描记的第6h分别给予腹腔注射不同剂量的NBI27914、阿托品和等量生理盐水。结果：与基础水平相比,腹腔注射任何剂量NBI27914使幼年大鼠快眼动（REM）睡眠均显著减少,同时伴随着非快眼动（NREM）睡眠的增加,而生理盐水组无显著变化;阿托品组的大鼠REM睡眠也显著减少,伴随的是觉醒的增加。结论：阻断CRFR1受体可以剥夺幼鼠的REM睡眠。     

NBI及AFE在喉癌早期诊断中的应用

研究表明,喉癌的早期诊断、及时治疗不仅可以提高治愈率,而且也减少了患者的手术创伤和经济负担。积极开展喉癌的早期诊断研究具有重要的临床和社会意义。发现早期喉癌常规方法主要有电子喉镜、纤维喉镜、颈部CT及MRI检查,但并不能明显有效提高早期诊断率。而窄带成像(narrow band imaging,NBI)及自体荧光内镜(autofluorescence endoscopy AFE)是近几年用于喉癌早期诊断的两种新颖的内镜技术。NBI是一种通过变窄光波的波长,使粘膜上皮内乳头样毛细血管袢及粘膜下静脉的结构形成鲜明的对比,从而提高组织表面细微结构的对比度,便于发现病灶。而AFE技术是一种利用自发荧光聚集于病变组织的某个区域产生的差异强度,来区别正常组织与肿瘤性病变,从而用于肿瘤的早期诊断及识别癌前病变。因此,对NBI及AFE的进一步研究及认识对喉癌早期诊断提供非常重要的临床应用价值。     

NICE分型在结直肠癌诊断中的意义

结直肠癌发病率高,早期诊断困难,威胁着全世界人民的健康。近几年窄带成像技术(Narrow Band Imaging,NBI)发展迅速,大大改善了结直肠镜下图像的清晰程度,NBI辅助下的结直肠镜检查在早期诊治肠癌方面具有重要意义。为了进一步优化结直肠镜的诊断效能,多种NBI辅助的结直肠镜下的结直肠病变分型被提出。针对病变的表面形态、颜色分布以及微血管的形态与走形,Sano分型、Jikei分型、Showa分型、Hiroshima分型和NICE(Narrow-Band Imaging International Colorectal Endoscopic)分型相继被提出。其中NICE分型以良好的学习曲线和诊断效能被广泛认可和应用。     

Preparation of DNA substrates for in vitro mismatch repair

Analyses in vitro of correction of DNA mismatches have been pivotal in biochemical dissection of mismatch repair pathways. However, the complex procedures needed to prepare DNA substrates for mismatch repair have posed substantial barriers to investigators who wish to pursue such analyses. Here we describe a simple, efficient way to prepare a variety of mismatched DNA substrates. We use in our procedure high-copy-number pUC19-derived plasmids, and a newly commercially available endonuclease N.BstNBI that makes site-specific single-strand nicks. The ability to prepare large substrate quantities in a relatively short time and to construct wider ranges of different mismatches in various sequence contexts will facilitate future research. Supported in part by NIH grant ESO94848 to J.B.H. This is Technical Report No. 11680 from the Oregon Agricultural Experiment Station