Hepatocyte growth factor induces redistribution of p21(CIP1) and p27(KIP1) through ERK-dependent p16(INK4a) up-regulation, leading to cell cycle arrest at G1 in HepG2 hepatoma cells

Hepatocyte growth factor (HGF) has an anti-proliferative effect on many types of tumor cell lines and tumors in vivo. We found previously that inhibition of HGF-induced proliferation in HepG2 hepatoma cells is caused by cell cycle arrest at G1 through a high intensity ERK signal, which represses Cdk2 activity. To examine further the mechanisms of G1 arrest by HGF, we analyzed the Cdk inhibitor p16(INK4a), which has an anti-proliferative function through cell cycle arrest at G1. We found that HGF treatment drastically increased endogenous p16 levels. Knockdown of p16 with small interfering RNA reversed the arrest, indicating that the induction of p16 is required for G1 arrest by HGF. Analysis of the promoter of the human p16 gene identified the proximal Ets-binding site as a responsive element for HGF, and this responded to the high intensity ERK signal. HGF treatment of the cells led to a redistribution of p21(CIP1) and p27(KIP1) from Cdk4 to Cdk2. The redistribution was blocked by the knockdown of p16 with small interfering RNA, which restored the Cdk2 activity repressed by HGF, demonstrating the requirement of p16 induction for the redistribution and eventual repression of Cdk2 activity. Our results reveal a signaling pathway for G1 arrest induced by HGF.     

Signals from intra-abdominal fat modulate insulin and leptin sensitivity through different mechanisms: neuronal involvement in food-intake regulation

Intra-abdominal fat accumulation is involved in development of the metabolic syndrome, which is associated with insulin and leptin resistance. We show here that ectopic expression of very low levels of uncoupling protein 1 (UCP1) in epididymal fat (Epi) reverses both insulin and leptin resistance. UCP1 expression in Epi improved glucose tolerance and decreased food intake in both diet-induced and genetically obese mouse models. In contrast, UCP1 expression in Epi of leptin-receptor mutant mice did not alter food intake, though it significantly decreased blood glucose and insulin levels. Thus, hypophagia induction requires a leptin signal, while the improved insulin sensitivity appears to be leptin independent. In wild-type mice, local-nerve dissection in the epididymis or pharmacological afferent blockade blunted the decrease in food intake, suggesting that afferent-nerve signals from intra-abdominal fat tissue regulate food intake by modulating hypothalamic leptin sensitivity. These novel signals are potential therapeutic targets for the metabolic syndrome.     

Cold exposure suppresses serum adiponectin levels through sympathetic nerve activation in mice

Objective: Several lines of evidence suggest important roles for adiponectin in glucose and lipid metabolism and atherosclerosis. However, the mechanisms regulating serum adiponectin levels and adiponectin production are still not completely understood. Our aim was to determine whether adiponectin synthesis is physiologically regulated by the sympathetic nervous system (SNS). Research Methods and Procedures: Mice were exposed to cold (4 °C) for 12 hours and for 24 hours with or without inhibition of noradrenaline synthesis or pan‐β adrenergic function, followed by measurement of serum adiponectin concentrations and levels of adiponectin and uncoupling protein (UCP) 1 expressions in various white adipose tissues (WATs). Results: Cold exposure significantly reduced serum adiponectin concentrations without changing body weights or WAT sizes in either subcutaneous or intra‐abdominal fat tissues. The serum adiponectin reduction was associated with a decrease in adiponectin mRNA expression in subcutaneous, epididymal, and mesenteric fat tissues. In these adipose tissues, UCP1 expression was markedly enhanced, suggesting SNS activation in these tissues. Administration of α‐methyl‐p‐tyrosine or a combination of SR59230A and propranolol reversed the cold‐exposure‐induced decreases in serum adiponectin concentrations and adiponectin mRNA expression in these tissues. In contrast, in retroperitoneal fat, the effects of cold exposure on adiponectin and UCP1 expressions were strikingly weak but were not reversed by SNS inhibitors. Discussion: SNS physiologically regulates serum adiponectin levels and adiponectin synthesis in WATs in vivo, although responsiveness to SNS stimulation differs markedly among WATs. Sympathetic activation might be involved in development of the metabolic syndrome by modulation of serum adiponectin concentrations.     

Therapeutic potential of in utero mesenchymal stem cell (MSCs) transplantation in rat foetuses with spina bifida aperta

Neural tube defects (NTDs) are complex congenital malformations resulting from incomplete neurulation in embryo. Despite surgical repair of the defect, most of the patients who survive with NTDs have a multiple system handicap due to neuron deficiency of the defective spinal cord. In this study, we successfully devised a prenatal surgical approach and transplanted mesenchymal stem cells (MSCs) to foetal rat spinal column to treat retinoic acid induced NTDs in rat. Transplanted MSCs survived, grew and expressed markers of neurons, glia and myoblasts in the defective spinal cord. MSCs expressed and perhaps induced the surrounding spinal tissue to express neurotrophic factors. In addition, MSC reduced spinal tissue apoptosis in NTD. Our results suggested that prenatal MSC transplantation could treat spinal neuron deficiency in NTDs by the regeneration of neurons and reduced spinal neuron death in the defective spinal cord.     

金华猪遗传结构及其与太湖猪遗传分化的研究

本研究利用65个微卫星标记结合荧光标记检测技术, 对金华猪I系、II系、III系共271个个体以及嘉兴黑猪、中梅山猪、小梅山猪和二花脸猪等4个太湖猪品种和嵊县花猪各30头的基因型进行了检测, 统计分析了金华猪各品系的遗传结构及各猪种群间的遗传分化。结果显示: 金华猪品系间具有丰富的遗传变异, 平均有效等位基因数以金华猪I系最高, 为3.5; 其次是II系和III系, 分别是2.8和2.5, 金华猪3个品系的平均多态信息含量均高于0.5; I、II、III系的平均观察杂合度分别是0.381、0.399和0.442。金华猪3个品系偏离Hardy-Weinberg平衡的程度不一:I系偏离较大, III系次之, II系相对较小。分析认为金华猪各品系存在一定程度的近交, 品系间存在不同的等位基因。遗传分化结果显示: 金华猪II系和III系间遗传分化相对较小(F_ST＝0.1883), 但它们与I系间的遗传分化较大, F_ST值分别是0.3663和0.3619。同时, 金华猪各品系与太湖猪的遗传关系较近, 其中与中梅山猪群体遗传分化相对较小, F_ST值分别为0.3581、0.3560和0.3572。而金华猪各品系与嵊县花猪的遗传分化最大, F_ST值分别为0.4499, 0.4654和0.4801, 由此可见, 金华猪不同于其他浙江省地方品种, 有着独立的起源和驯化进程。     

Changes in landscape pattern of alpine wetlands on the Zoige Plateau in the past four decades

Based on RS, GIS and Apack software, the indices of landscape pattern such as landscape area index, landscape diversity index and landscape fragmentation index were chosen in order to describe changes in the spatial pattern of alpine wetland landscape on the Zoige Plateau during 1966–2000. Results showed that alpine wetland landscape was characteristic of marsh wetlands, which had the biggest patch number and the largest area. The alpine wetland landscape had higher spatial heterogeneity. The largest area appeared in Zoige County with the highest wetland ratio; comparatively, Aba County and Luqu County had much lower wetland ratio. The total area of alpine wetland landscape decreased rapidly during 1966–1986, but it began to increase after 1986. The wetland landscape area shrank by 59857.83 hm² during 1966–2000. The alpine wetland landscape showed the characteristics of concentrated distribution in the past four decades, with higher convergence and dominance indices. The centroid of wetland landscape moved 12.54 km in the northwest direction firstly, 11.33 km in the southeast direction, and then 1.1 km in the north direction.     

METTL3-mediated mRNA N6-methyladenosine is required for oocyte and follicle development in mice

Proper follicle development is very important for the production of mature oocytes, which is essential for the maintenance of female fertility. This complex biological process requires precise gene regulation. The most abundant modification of mRNA, N⁶-methyladenosine (m⁶A), is involved in many RNA metabolism processes, including RNA splicing, translation, stability, and degradation. Here, we report that m⁶A plays essential roles during oocyte and follicle development. Oocyte-specific inactivation of the key m⁶A methyltransferase Mettl3 with Gdf9-Cre caused DNA damage accumulation in oocytes, defective follicle development, and abnormal ovulation. Mechanistically, combined RNA-seq and m⁶A methylated RNA immunoprecipitation sequencing (MeRIP-seq) data from oocytes revealed, that we found METTL3 targets Itsn2 for m⁶A modification and then enhances its stability to influence the oocytes meiosis. Taken together, our findings highlight the crucial roles of mRNA m⁶A modification in follicle development and coordination of RNA stabilization during oocyte growth.Subject terms: Oogenesis, Infertility     

A new PCR-based marker on chromosome 4AL for resistance to pre-harvest sprouting in wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.)

Pre-harvest sprouting (PHS) is a complex trait controlled by multiple genes with strong interaction between environment and genotype that makes it difficult to select breeding materials by phenotypic assessment. One of the most important genes for pre-harvest sprouting resistance is consistently identified on the long arm of chromosome 4A. The 4AL PHS tolerance gene has therefore been targeted by Australian white-grained wheat breeders. A new robust PCR marker for the PHS QTL on wheat chromosome 4AL based on candidate genes search was developed in this study. The new marker was mapped on 4AL deletion bin 13-0.59-0.66 using 4AL deletion lines derived from Chinese Spring. This marker is located on 4AL between molecular markers Xbarc170 and Xwg622 in the doubled-haploid wheat population Cranbrook × Halberd. It was mapped between molecular markers Xbarc170 and Xgwm269 that have been previously shown to be closely linked to grain dormancy in the doubled haploid wheat population SW95-50213 × Cunningham and was co-located with Xgwm269 in population Janz × AUS1408. This marker offers an additional efficient tool for marker-assisted selection of dormancy for white-grained wheat breeding. Comparative analysis indicated that the wheat chromosome 4AL QTL for seed dormancy and PHS resistance is homologous with the barley QTL on chromosome 5HL controlling seed dormancy and PHS resistance. This marker will facilitate identification of the gene associated with the 4A QTL that controls a major component of grain dormancy and PHS resistance.     

稻田杂草稻发生情况调查

2007年杂草稻在江苏省海安县各类稻田中的发生率达44．35％,平均每公顷穴数和穗数分别为1023穴和10796穗,最高田块分别达12万穴和180万穗,已成为水稻生产上的潜在威胁。杂草稻在不同稻田的发生程度表现以下趋势：轻-中壤〉砂-轻壤〉重壤土稻田、直播〉抛栽〉机插〉手栽稻田、非统一供种〉统一供种稻田。