Loss of IGF2 imprinting: a potential marker of colorectal cancer risk

Loss of imprinting (LOI), an epigenetic alteration affecting the insulin-like growth factor II gene (IGF2), is found in normal colonic mucosa of about 30% of colorectal cancer (CRC) patients, but it is found in only 10% of healthy individuals. In a pilot study to investigate the utility of LOI as a marker of CRC risk, we evaluated 172 patients at a colonoscopy clinic. The adjusted odds ratio for LOI in lymphocytes was 5.15 for patients with a positive family history [95% confidence interval (95% CI), 1.70 to 16.96; probability P = 0.002], 3.46 for patients with adenomas (95% CI, 1.14 to 11.37; P = 0.026), and 21.7 for patients with CRC (95% CI, 3.48 to 153.6; P = 0.0005). LOI can be assayed with a DNA-based blood test, and it may be a valuable predictive marker of an individual's risk for CRC.     

Loss of sex discrimination and male-male aggression in mice deficient for TRP2

The mouse vomeronasal organ (VNO) is thought to mediate social behaviors and neuroendocrine changes elicited by pheromonal cues. The molecular mechanisms underlying the sensory response to pheromones and the behavioral repertoire induced through the VNO are not fully characterized. Using the tools of mouse genetics and multielectrode recording, we demonstrate that the sensory activation of VNO neurons requires TRP2, a putative ion channel of the transient receptor potential family that is expressed exclusively in these neurons. Moreover, we show that male mice deficient in TRP2 expression fail to display male-male aggression, and they initiate sexual and courtship behaviors toward both males and females. Our study suggests that, in the mouse, sensory activation of the VNO is essential for sex discrimination of conspecifics and thus ensures gender-specific behavior.     

Enhanced CpG mutability and tumorigenesis in MBD4-deficient mice

The mammalian protein MBD4 contains a methyl-CpG binding domain and can enzymatically remove thymine (T) or uracil (U) from a mismatched CpG site in vitro. These properties suggest that MBD4 might function in vivo to minimize the mutability of 5-methylcytosine by removing its deamination product from DNA. We tested this hypothesis by analyzing Mbd4-/- mice and found that the frequency of of C --> T transitions at CpG sites was increased by a factor of three. On a cancer-susceptible Apc(Min/+) background, Mbd4-/- mice showed accelerated tumor formation with CpG --> TpG mutations in the Apc gene. Thus MBD4 suppresses CpG mutability and tumorigenesis in vivo.     

Regulation of spontaneous intestinal tumorigenesis through the adaptor protein MyD88

Inflammation is increasingly recognized as an important component of tumorigenesis, although the mechanisms and pathways involved are not well understood. Tumor development is regulated by products of several modifier genes, but instructions for their tumor-specific expression are currently unknown. We show that the signaling through the adaptor protein MyD88 has a critical role in spontaneous tumor development in mice with heterozygous mutation in the adenomatous polyposis coli (APC) gene. We found that MyD88-dependent signaling controls the expression of several key modifier genes of intestinal tumorigenesis and has a critical role in both spontaneous and carcinogen-induced tumor development. This study thus reveals the important role of an innate immune signaling pathway in intestinal tumorigenesis.     

褪黑素在高龄小鼠卵母细胞体外成熟中的促进作用

为探究褪黑素对高龄小鼠卵母细胞体外成熟的影响,以10～12月龄的ICR雌鼠为试验材料,分别将不同浓度的褪黑素(0、10-4、10-6、10-8 mol·L-1)添加至体外成熟培养液中,通过判断卵母细胞的成熟率、受精率及囊胚率筛选出褪黑素最适宜的添加浓度,并通过线粒体分布检测、活性氧水平检测和细胞早期凋亡水平检测,分析褪...     

印迹基因Igf2r的甲基化进程与兔卵母细胞直径关系的研究

本研究以3～4月龄兔为研究对象,收集卵母细胞,按直径将其划分为75～95μm、95～105μm、105～115μm3个组,通过亚硫酸盐测序法检测了印迹基因Igf2r的甲基化程度,并运用RT-PCR对Dnmt1、Dnmt3a、Dnmt3b、Dnmt3l、Lsh的表达量进行检测。结果表明,在3组卵母细胞内,Igf2r的甲基化比率依次为15.7%、60%、91.5%,甲基化程度随卵母细胞直径的增加而不断提高。     

Vasoactive intestinal peptide alters membrane potential and cyclic nucleotide levels in retinal horizontal cells

Vasoactive intestinal peptide stimulated the synthesis of adenosine 3',5'-monophosphate in fractions of isolated carp horizontal cells. When applied extracellularly to isolated and cultured horizontal cells, the peptide also induced a slow depolarization (30 to 40 millivolts) accompanied by a decrease in membrane resistance. However, analogs of adenosine 3',5'-monophosphate applied extracellularly or intracellularly, and forscolin applied extracellularly, had no effect on the membrane potential of cultured horizontal cells, indicating that the induced depolarization was not related to the accumulation of adenosine 3',5'-monophosphate in these cells.     

Ornithine decarboxylase is important in intestinal mucosal maturation and recovery from injury in rats

A transient increase in ornithine decarboxylase activity and polyamine biosynthesis occurs in the intestinal mucosa of the newborn rat in the third week after birth. During this period, there is a rapid conversion of the mucosa from a fetal to a mature adult status. A similar increase in ornithine decarboxylase activity also accompanies the rapid recovery of the mucosa 1 week after an injury is induced by chemotherapy in adult rats. In vivo, alpha-difluoromethyl ornithine, a highly selective, enzyme-activated, irreversible inhibitor, suppresses these increases in mucosal ornithine decarboxylase and delays both intestinal mucosal maturation and recovery from injury. Thus increased ornithine decarboxylase activity, with the resultant increase in polyamine content, may play an essential role in intestinal mucosal maturation and regeneration in the rat.     

溶葡萄球菌素对小鼠肠道菌群结构的影响

【目的】研究溶葡萄球菌素对小鼠肠道菌群结构的影响,为溶葡萄球菌素的食用安全性评估提供参考。【方法】将24只7周龄ICR小鼠随机分为3个试验组和对照组,3个试验组分别提供溶葡萄球菌素含量为1 μg/mL（低剂量）、5 μg/mL（中剂量）和15 μg/mL（高剂量）的饮水10 mL/(只·d),于饮用第0(饮用前),1,3,7,14,21天以及停饮第3,7,14天采集小鼠新鲜粪便样品,采用PCRDGGE技术检测小鼠肠道菌群结构变化。【结果】DGGE图谱显示,各组小鼠试验前后不同时间肠道内的优势菌种相似;经UPGMA聚类分析,与对照组相比,低、中剂量溶葡萄球菌素对小鼠肠道菌群结构影响较小,高剂量溶葡萄球菌素对小鼠肠道菌群结构影响较大;差异性分析结果显示,试验组饮用各阶段带谱与试验前的相似性与对照组相应阶段的相似性差异均不显著（P＞0.05）,溶葡萄球菌素低、中和高剂量组在停饮后第7天带谱与饮用前的相似性分别达到了70.8%,66.9%和63.8%;对图谱中主要条带进行测序和比对分析,结果表明,高剂量溶葡萄球菌素对小鼠肠道中Lactobacillus taiwanensis和1株不可培养细菌有促进作用。【结论】溶葡萄球菌素对小鼠肠道菌群结构的改变具有浓度依赖性,且停饮后小鼠肠道菌群结构有一定程度的恢复。     

Prevention of Brca1-mediated mammary tumorigenesis in mice by a progesterone antagonist

Women with mutations in the breast cancer susceptibility gene BRCA1 are predisposed to breast and ovarian cancers. Why the BRCA1 protein suppresses tumor development specifically in ovarian hormone-sensitive tissues remains unclear. We demonstrate that mammary glands of nulliparous Brca1/p53-deficient mice accumulate lateral branches and undergo extensive alveologenesis, a phenotype that occurs only during pregnancy in wild-type mice. Progesterone receptors, but not estrogen receptors, are overexpressed in the mutant mammary epithelial cells because of a defect in their degradation by the proteasome pathway. Treatment of Brca1/p53-deficient mice with the progesterone antagonist mifepristone (RU 486) prevented mammary tumorigenesis. These findings reveal a tissue-specific function for the BRCA1 protein and raise the possibility that antiprogesterone treatment may be useful for breast cancer prevention in individuals with BRCA1 mutations.     

伴大豆球蛋白胃蛋白酶水解肽对小鼠免疫功能及肠道内环境的影响

90只清洁级昆明系小鼠,随机分为5组,预饲基础日粮一周后,各组分别灌胃：生理盐水（对照组,CK）、伴大豆球蛋白液（Ⅰ）、伴大豆球蛋白胃蛋白酶水解液（Ⅱ）、伴大豆球蛋白胃蛋白酶水解液提纯B组分（Ⅲ）、伴大豆球蛋白盐酸水解液（Ⅳ）,除对照组外,各处理组灌胃液体总含氮量相等,实验期21d。结果显示：与对照组相比,第3周Ⅱ组和Ⅲ组每只小鼠日均采食量较对照组显著下降,分别降低8．75％（P〈0．05）和9．28％（P〈0．05）;Ⅲ组小鼠胸腺相对质量极显著下降,降低了10．40％（P〈0．01）,肠黏膜sIgA水平提高45．49％（P〈0．01）,大肠食糜中6-磷酸果糖酮糖酶（F6PPK）活性极显著提高（P〈0．01）,pH值下降了1．80％（P〈0．05）,肠球菌数显著下降（P〈0．05）。实验结果表明水解肽作为腔内信号分子可能与肠道内相关因素有着多层次调控关系,共同促进宿主的健康。     

Partial inversion of the initiation-promotion sequence of multistage tumorigenesis in the skin of NMRI mice

Alterations in NMRI mouse skin induced by the phorbol ester 12-O-tetradecanoylphorbol-13-acetate in "stage I of tumor promotion" are slowly reversible, and this reversibility has a half-time of 10 to 12 weeks. The tumor response observed in the course of an initiation-promotion experiment in vivo is independent of whether stage I of promotion occurs before or after initiation. Since the time interval between treatment with the promoter, and subsequent initiation can be extended up to at least 6 weeks, an enhancement of initiation because of promoter-induced cellular DNA synthesis seems to be unlikely. This result may be inconsistent with the two-stage model of tumor promotion because it indicates that in skin the existence of initiated cells is not required for the induction of cellular alterations that are essential for the stage of skin tumorigenesis called stage I of promotion.     

Early-life blockade of the 5-HT transporter alters emotional behavior in adult mice

Reduced serotonin transporter (5-HTT) expression is associated with abnormal affective and anxiety-like symptoms in humans and rodents, but the mechanism of this effect is unknown. Transient inhibition of 5-HTT during early development with fluoxetine, a commonly used serotonin selective reuptake inhibitor, produced abnormal emotional behaviors in adult mice. This effect mimicked the behavioral phenotype of mice genetically deficient in 5-HTT expression. These findings indicate a critical role of serotonin in the maturation of brain systems that modulate emotional function in the adult and suggest a developmental mechanism to explain how low-expressing 5-HTT promoter alleles increase vulnerability to psychiatric disorders.     

两色金鸡菊乙酸乙酯提取物对2型糖尿病小鼠肠道菌群的影响

为研究两色金鸡菊乙酸乙酯提取物对糖尿病小鼠肠道菌群的影响,初步判断它们之间的关系,收集正常组、模型组、二甲双胍对照组以及两色金鸡菊乙酸乙酯提取物高、中、低剂量组粪便样本,采用实时荧光定量PCR技术检测粪便样本中球形梭菌与多形拟杆菌的水平;运用Pearson分析,对目标菌属水平与各组小鼠的空腹血糖(FBG)做相关性分析。结果表明:(1)与正常组比较,模型组中球形梭菌与多形拟杆菌水平均增高(P=0.017,P=0.002);(2)两色金鸡菊乙酸乙酯提取物低剂量组与模型组相比,球形梭菌与多形拟杆菌的水平下降,且有统计学差异(2周:P=0.010,P=0.002;4周:P=0.001,P=0.002);(3)球形梭菌水平、多形拟杆菌水平均与FBG水平呈正相关。可见2型糖尿病小鼠肠道菌群球形梭菌与多形拟杆菌水平较正常组增高;两色金鸡菊乙酸乙酯提取物可降低模型组群球形梭菌与多形拟杆菌水平。     

牛磺酸对断奶小鼠小肠发育的影响

为研究牛磺酸对断奶小鼠小肠结构和功能的影响,采用单因子设计,48只体重为(12±2)g的昆明小鼠随机分为6组,每组8只,雌雄各半,分为对照组和试验组,对照组饮用普通自来水,试验组在饮水中添加2.5、5.0、10.0、20.0和40.0g/L牛磺酸,试验期31d。观察牛磺酸对小肠和肝脏重量,空肠绒毛形态、细胞凋亡、小肠核酸和蛋白质含量,以及血清指标的影响。结果表明:1)饮水中添加牛磺酸可显著提高小鼠小肠绝对重量和相对重量(P0.05),且呈现二次曲线关系(PQ0.05);可线性提高小肠绝对长度、肝脏绝对重量和相对重量(PL0.05)。2)2.5、5.0和10.0g/L组与对照组相比,显著降低空肠肠腺(肠隐窝)深度(P0.05);回肠Prot/DNA随牛磺酸添加量提高而线性提高(PL0.05)。3)与对照组比较,5.0g/L添加组的细胞凋亡指数显著降低(P0.05),但40g/L添加组有升高的趋势。4)随着牛磺酸添加量的提高,回肠MDA含量有降低的趋势(P=0.07),空肠GSHPx呈现先升高再降低的二次曲线关系(PQ0.05),但不影响血清IGF-I水平。研究结果表明,饮水添加中低浓度的牛磺酸可提高小鼠小肠长度和重量,降低肠隐窝深度,减少细胞凋亡。     

特异性敲除肠道miR-146a对小鼠肠道菌群的影响

【目的】miR-146a作为抑炎因子,仍然不清楚其是否参与宿主与微生物间的互作,进而影响肠道稳态,因此本文旨在研究miR-146a对小鼠肠道菌群的影响。【方法】以肠道miR-146a特异性敲除小鼠(CKO鼠)及对照小鼠(Flox鼠)为研究对象,利用16S rRNA高通量测序法检测2组空肠段的微生物菌群分布。【结果】测序共获得1 134个用于物种分类的OTUs,包括37门、80纲、161目、198科、261属、117种的细菌;Flox组和CKO组小鼠的空肠微生物中共有46个相同的OTUs;各组肠道微生物中厚壁菌门Firmicutes、拟杆菌门Bacteroidota、疣微菌门Verrucomicrobiota、变形菌门Proteobacteria和脱硫杆菌门Desulfobacterota是优势菌门;2组肠道微生物群落组成整体相似,但CKO组梭状芽孢杆菌纲Clostridia的平均相对丰度高于Flox组(P=0.067),毛螺菌目Lachnospirales平均相对丰度显著高于Flox组(P<0.05),其他层级组成无显著差异。【结论】miR-146a敲除可改变宿主肠道梭状芽孢杆菌...     

大豆异黄酮转化菌株在ICR小鼠体内定植研究

健康ICR小鼠肠道菌群对大豆异黄酮有稳定转化作用。本研究选用合适抗生素对健康ICR小鼠进行连续灌服,使ICR小鼠在不影响其正常生理活动状况下完全丧失对大豆异黄酮的转化能力,并对该类完全丧失转化能力的ICR小鼠灌服大豆异黄酮还原菌株Niu-O16的微胶囊,用高效液相色谱检测菌株Niu-O16在ICR小鼠体内定植情况。结果表明:用420mg/mL浓度的阿奇霉素连续灌服1周,可使ICR小鼠完全丧失对大豆异黄酮的转化能力,停药后20d左右,ICR小鼠恢复其对大豆异黄酮的转化能力;经抗生素处理的ICR小鼠在停止灌菌后,2周内可检测到其粪便菌群具有与菌株Niu-O16完全相同的转化作用,而同样经抗生素处理未灌菌的对照则未检测到转化活性,表明菌株Niu-O16已在ICR小鼠体内进行定植。     

发酵乳对肥胖小鼠肝脂代谢和肠道菌群的影响

为分析发酵乳对肥胖小鼠肝脂代谢和肠道菌群的影响,本研究将60只C57BL/6小鼠随机分为5组：正常对照组（CON）、模型组（MD）、易善复组（YG）、低浓度发酵乳干预组（LFG）和高浓度发酵乳干预组（HFG）。肥胖模型造模成功后进行为期6周的灌胃,试验结束测量小鼠的空腹体重和身长,计算LEE’S指数,采血检测生化指标,采集肝脏组织观察病理变化;采集小鼠盲肠内容物进行肠道菌群测序。结果表明：1）与CON相比,MD小鼠体重、LEE’S指数、ALT和AST均极显著提升（P<0.01）,表明肥胖模型建立成功。2）与MD相比,发酵乳干预组小鼠体重、LEE’S指数、ALT和TC均极显著下降（P<0.01）,其中AST和TG显著下降（P<0.05）,同时发酵乳和易善复组小鼠肝组织脂肪变性有明显好转,基本没有明显的脂肪空泡。表明发酵乳可以抑制肥胖小鼠体重的增加,改善肥胖小鼠的肝脂代谢功能。3）与MD相比干预组小鼠肠道菌群的多样性水平有一定程度的恢复,尤其是厚壁菌门和拟杆菌门。表明发酵乳对于小鼠肠道菌群多样性的恢复有一定的作用。综上,本研究阐明了发酵乳对肥胖小鼠肝脂代谢和肠道菌群的影响,为功能型发酵乳的开发提供理论依据。