Characterization of newly established colorectal cancer cell lines: correlation between cytogenetic abnormalities and allelic deletions associated with multistep tumorigenesis

We have established a series of 20 colorectal cancer cell lines and performed cytogenetic and RFLP analyses to show that the recurrent genetic abnormalities of chromosomes 1, 5, 17 and 18 associated with multistep tumorigenesis in colorectal cancer, and frequently detected as recurrent abnormalities in primary tumours, are also retained in long-term established cell lines. Earlier studies by us and other investigators showed that allelic losses of chromosomes 1 and 17 in primary colorectal cancers predicted poorer survival for the patients (P = 0.03). We utilized the cell lines to identify specific chromosomal sites or gene(s) on chromosomes 1 and 17 which confer more aggressive phenotype. Cytogenetic deletions of chromosome 1p were detected in 14 out of the 20 (70%) cell lines, whereas allelic deletions for 1p using polymorphic markers were detected in 13 out of 18 (72%) informative cell lines for at least one polymorphic marker. We have performed Northern blotting, immunohistochemical staining (p53 mRNA, protein) and RFLP analysis using several probes including p53 and nm23. RFLP analysis using a total of seven polymorphic markers located on 17p and 17q arms showed allelic losses aroundthe p53 locus in 16 out of the 20 cell lines (80%), four of which were losses of thep53 locus itself. In addition, seven cell lines (out of nine informative cases) also showed losses of thenm23 gene, four with concurrent losses of thep53 locus, while the remaining three were homozygous. In addition, five out of seven cell lines withnm23 deletions were derived from hepatic metastatic tumours, and one cell line was obtained from recurrent tumour. A comparison between allelic deletions of 1p and functional loss ofnm23 gene revealed a close association between these two events in cell lines derived from hepatic metastasis. Following immunohistochemical staining, nine out of the twenty cell lines showed high levels (25–80%) of mutant p53, four showed intermediate levels (>20%), and seven had undetectable levels of the protein. Of these seven, four showed complete absence of mRNA. Of the remaining three cell lines one showed aberrant mRNA due to germline rearrangement of thep53 gene, whereas in two cell lines normal levels of mRNA were present. Nineteen of the 20 cell lines had normal germline configurations for thep53 gene, while one showed a rearrangement. These data suggest that functional loss ofp53 andnm23 genes accomplished by a variety of mechanisms may be associated with poor prognosis and survival. In addition, concurrent deletions of chromosome regions 17p, 17q and 1p were closely associated with high-stage hepatic metastatic disease. These cell lines with well-characterized genetic alterations and known clinical history provide an invaluable source of material for various biological and clinical studies relating to multistep colorectal tumorigenesis.     

Étude électrophorétique de l'hybride expérimental entre Papio anubis et P. cynocephalus

Hybrids between Papio anubis and P. cynocephalus occur in nature. We have studied laboratory bred hybrids, together with their backcrossed offspring. Electrophoretic experiments on the proteins, serum enzymes and erythrocytes obtained from both hybrids and parents lead to three conclusions. First, both parents and hybrids possess proteins having identical mobilities; this case is the most frequent in the 23 loci studied. Second, individual polymorphism caused by, for example, carbonic anhydrase I or transferrin in one or both parents results in the appearance of a codominant phenotype in the hybrid, when the parental components are distinguishable. Third, when other enzymes such as albumin, alkaline phosphatase, haemoglobin and the serum esterases have particular electrophoretic mobilities in the parents, then the hybrid is again codominant; a particularly interesting case among the esterases is that of pseudocholinesterase, where the paternal component is partially repressed in the hybrid. The hybrid than exhibits the phenomenon of allelic repression of at least one gene, which confirms the taxonomic position of species of the two forms of Papio studied.     

Context-Dependent Sensitivity to Mutations Disrupting the Structural Integrity of Individual EGF Repeats in the Mouse Notch Ligand DLL1

The highly conserved Notch-signaling pathway mediates cell-to-cell communication and is pivotal for multiple developmental processes and tissue homeostasis in adult organisms. Notch receptors and their ligands are transmembrane proteins with multiple epidermal-growth-factor-like (EGF) repeats in their extracellular domains. In vitro the EGF repeats of mammalian ligands that are essential for Notch activation have been defined. However, in vivo the significance of the structural integrity of each EGF repeat in the ligand ectodomain for ligand function is still unclear. Here, we analyzed the mouse Notch ligand DLL1. We expressed DLL1 proteins with mutations disrupting disulfide bridges in each individual EGF repeat from single-copy transgenes in the HPRT locus of embryonic stem cells. In Notch transactivation assays all mutations impinged on DLL1 function and affected both NOTCH1 and NOTCH2 receptors similarly. An allelic series in mice that carried the same point mutations in endogenous Dll1, generated using a mini-gene strategy, showed that early developmental processes depending on DLL1-mediated NOTCH activation were differently sensitive to mutation of individual EGF repeats in DLL1. Notably, some mutations affected only somite patterning and resulted in vertebral column defects resembling spondylocostal dysostosis. In conclusion, the structural integrity of each individual EGF repeat in the extracellular domain of DLL1 is necessary for full DLL1 activity, and certain mutations in Dll1 might contribute to spondylocostal dysostosis in humans.     

The NAD-Booster Nicotinamide Riboside Potently Stimulates Hematopoiesis through Increased Mitochondrial Clearance

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Greater soil microbial biomass loss at low frequency of N addition in an Inner Mongolia grassland

低频率的氮添加使内蒙古草原土壤微生物生物量碳出现更大幅度下降 土壤微生物生物量在生物地球化学循环过程中至关重要,是土壤碳固持的前体物质。人为氮输入深刻地改变了草地土壤微生物生物量。然而,传统氮沉降模拟实验仅通过低频率的氮添加进行,与持续高频率的自然氮沉降相比,对土壤微生物生物量的影响可能存在差异。不同频率的氮添加对土壤微生物生物量的影响尚缺乏可靠的数据支撑。本研究通过在不同的氮添加速率(0–50 g N m−2 yr−1)下,控制氮添加频率(每年2次和12次),研究了土壤微生物生物量碳对不同氮添加频率的响应。研究结果表明,在两种氮添加频率下,随着施氮水平的提高,土壤微生物生物量碳逐渐降低。然而,在低施氮频率下,土壤微生物生物量的下降幅度更大,这说明传统的氮添加实验可能高估了氮沉降对土壤微生物生物量的影响。在低施氮频率下,土壤酸化、无机氮积累、碳氮失衡、地下净初级生产力分配减少和真菌细菌比例降低等情况加剧,导致微生物生物量出现较大幅度下降。在未来研究中,为可靠预测氮沉降对草地生态系统土壤微生物功能和碳循环的影响,不仅要考虑氮添加的剂量,还需要考虑氮添加的频率。     

LenkeⅡ型青少年特发性脊柱侧凸患者内固定冠状面失衡的相关因素分析

目的:探讨Lenke II型青少年特发性脊柱侧凸患者内固定冠状面失衡相关因素分析。方法:选取我院骨科已确诊为Lenke II青少年特发性脊柱侧凸患者60例,根据术前脊柱柔韧度、risser分级水平、支具支持治疗、椎体融合数、内固定系统选择等多方面影响因素对内固定后冠状面失平衡情况发生率进行分析评估,其数据结果应用统计学软件SPSS 17.0处理。结果:对术后患者评估比较,多种因素均可影响内固定后冠状面失平衡情况发生,表现为:脊柱柔韧度低、risser分级高、长时间支具佩戴、延长融合椎体范围、先进钉棒系统方案选择均可明显提高术后疗效,降低内固定后冠状面失平衡情况发生率(P0.05),其结果均有统计学意义。结论:临床上通过对患者脊柱柔韧度、risser分级情况评估来判断脊柱成熟度,并在术后积极支具辅助治疗,选用钉棒固定系统并延长融合椎体节段,可明显降低内固定后冠状面失平衡情况发生,提高远期随访疗效。     

双歧杆菌预防化疗后肠道菌群失调症的临床研究

探讨双歧杆菌对乳腺癌患者由于化疗导致肠道菌群失调的预防作用。2010年在我科住院的98例乳腺癌患者,在第一周期化疗后有48例出现肠道菌群失调,发生率为48.9%;在3周后的第二周期化疗前1d予患者口服双歧杆菌（双歧杆菌乳杆菌三联活菌片,一次4片,一日2次）,直至化疗后2周,肠道菌群失调例数减少到32例,发生率为32.7%。在前后两周期化疗期间监测血常规,粪便菌群失调检查。治疗周期出现菌群失调比例32.7%明显低于对照周期48.9%（P〈0.05）,双歧杆菌对乳腺癌患者化疗导致胃肠菌群失调有预防作用。