耐热性碱性磷酸酯酶作为非放射性标记物的研究

通过生物素与亲和素－酶复合物系统或地高辛与抗地高辛－酶复合物系统可把酶间接标记到探针上．Ｒｅｎｚ等通过不同的化学方法直接把酶标记到探针上［１～３］．耐热性碱性磷酸酯酶ＦＤ－ＴＡＰ（ｔｈｅｒｍｏｓｔａｂｌｅａｌｋａｌｉｎｅｐｈｏｓｐｈａｔａｓｅ）具有耐...     

我国特有植物银杉的资源、分布及其环境

我国银杉自1955年发现以来,陆续发现了它的新分布,至今已知在四省、区7个分布区的30多个分布点上,自然生长着银杉3200余株。它们是:广西花坪林区(越城岭南部)6个点有银杉1040抹,其中高5米以上的68株,1～5米的49株,1米以下的923株,最高21.1米,最大胸径83厘米;广西大瑶山4个点有银杉143株,其中高10米以上的94株,1～3米的3株,1米以下的46株,最高30米,最大胸径79.2厘米;湖南罗汉洞(越城岭北部)2个点有银杉58株,5米以上的有45株,最高18米,最大胸径43厘米;湖南罗霄山脉八面山有银杉707株,高1米以上的334株,最高24米,最大胸径46厘米;四川南川金佛山(大娄山东段北部)6个点有银杉527株,其中成年树400余株,最高17米,最大胸径50厘米;百枝山有银杉52株,最高8米,最大胸径27厘米;贵州道真沙河林区(大娄山东段南部)3处12个点有银杉729株,其中高5米以上的105株,2～5米的209株,1～2米的111株,1米以下的304株,最高18.8米,最大胸径48厘米;贵州桐梓白菁有银杉10株,高4～5米的2株,1～3米的6株,1米以下的2株。 银杉分布区范围自北纬24°5′～29°13′14″,东经107°10′～113°40′。其垂直分布自海拔940～1840米,金佛山银杉分布最高,为1600～1840米;罗汉洞银杉分布最低,为940～1060米。分布区内,年均温8.2°～17℃,绝对最低温     

Both low and high concentrations of staurosporine induce G1 arrest through down-regulation of cyclin E and cdk2 expression

Staurosporine has been reported to cause arrest of cells in G1 phase at low concentration and in G2 phase at high concentration. This raises the question of why the effects of staurosporine on the cell cycle depend on the applied concentration. In order to verify these multiple functions of staurosporine in Meth-A cells, we used cyclin E as a landmark of G1/S transition, cyclin B as a landmark of G2/M transition and MPM2 as a hallmark of M phase. We found that staurosporine arrested cells in G1 phase at a low concentration (20 nM) and in G2/M phase at a high concentration (200 nM). However, 200 nM staurosporine increased the expression of cyclin B and cdc2 proteins, suggesting that the cells progressed through the G2/M transition, and increased the expression of MPM2 protein, indicating that the cells entered M phase. Moreover, 200 nM staurosporine increased the expression of p53 and p21 proteins and inhibited the expression of cyclin E and cdk2 proteins, suggesting that the cells were arrested in the G1 phase of the next cycle. Morphological observation showed similar results as well. These data suggest that the G2/M accumulation induced by 200 nM staurosporine does not reflect G2 arrest, but rather results from M phase arrest, followed by progression from M phase to the G1 phase of the next cycle without cytokinesis, and finally arrest of the cells in G1 phase.     

The Orphan Nuclear Receptor NR4A1 Protects Pancreatic β-Cells from Endoplasmic Reticulum (ER) Stress-mediated Apoptosis

The role of NR4A1 in apoptosis is controversial. Pancreatic β-cells often face endoplasmic reticulum (ER) stress under adverse conditions such as high free fatty acid (FFA) concentrations and sustained hyperglycemia. Severe ER stress results in β-cell apoptosis. The aim of this study was to analyze the role of NR4A1 in ER stress-mediated β-cell apoptosis and to characterize the related mechanisms. We confirmed that upon treatment with the ER stress inducers thapsigargin (TG) or palmitic acid (PA), the mRNA and protein levels of NR4A1 rapidly increased in both MIN6 cells and mouse islets. NR4A1 overexpression in MIN6 cells conferred resistance to cell loss induced by TG or PA, as assessed by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, and TUNEL assays indicated that NR4A1 overexpression also protected against ER stress-induced apoptosis. This conclusion was further confirmed by experiments exploiting siRNA to knockdown NR4A1 expression in MIN6 cells or exploiting NR4A1 knock-out mice. NR4A1 overexpression in MIN6 cells reduced C/EBP homologous protein (CHOP) expression and Caspase3 activation induced by TG or PA. NR4A1 overexpression in MIN6 cells or mouse islets resulted in Survivin up-regulation. A critical regulatory element was identified in Survivin promoter (−1872 bp to −1866 bp) with a putative NR4A1 binding site; ChIP assays demonstrated that NR4A1 physically associates with the Survivin promoter. In conclusion, NR4A1 protects pancreatic β-cells against ER stress-mediated apoptosis by up-regulating Survivin expression and down-regulating CHOP expression, which we termed as “positive and negative regulation.”     

βII spectrin (SPTBN1): biological function and clinical potential in cancer and other diseases

βII spectrin, the most common isoform of non-erythrocyte spectrin, is a cytoskeleton protein present in all nucleated cells. Interestingly, βII spectrin is essential for the development of various organs such as nerve, epithelium, inner ear, liver and heart. The functions of βII spectrin include not only establishing and maintaining the cell structure but also regulating a variety of cellular functions, such as cell apoptosis, cell adhesion, cell spreading and cell cycle regulation. Notably, βII spectrin dysfunction is associated with embryonic lethality and the DNA damage response. More recently, the detection of altered βII spectrin expression in tumors indicated that βII spectrin might be involved in the development and progression of cancer. Its mutations and disorders could result in developmental disabilities and various diseases. The versatile roles of βII spectrin in disease have been examined in an increasing number of studies; nonetheless, the exact mechanisms of βII spectrin are still poorly understood. Thus, we summarize the structural features and biological roles of βII spectrin and discuss its molecular mechanisms and functions in development, homeostasis, regeneration and differentiation. This review highlight the potential effects of βII spectrin dysfunction in cancer and other diseases, outstanding questions for the future investigation of therapeutic targets. The investigation of the regulatory mechanism of βII spectrin signal inactivation and recovery may bring hope for future therapy of related diseases.     

四川盐源盆地哺乳类化石及其意义

本文简述盐源盆地上新世和更新世晚期两个不同时代的哺乳动物化石十余种,据之修正了该盆地晚新生代沉积的时代和对比关系.     

Design, synthesis, and evaluation of small molecule Hsp90 probes

A number of compounds from different chemical classes are known to bind competitively to the ATP-pocket of Hsp90 and inhibit its chaperone function. The natural product geldanamycin was the first reported inhibitor of Hsp90 and since then synthetic inhibitors from purine, isoxazole and indazol-4-one chemical classes have been discovered and are currently or soon to be in clinical trials for the treatment of cancer. In spite of a similar binding mode to Hsp90, distinct biological profiles were demonstrated among these molecules, both in vitro and in vivo. To better understand the molecular basis for these dissimilarities, we report here the synthesis of chemical tools for three Hsp90 inhibitor classes. These agents will be useful for probing tumor-by-tumor the Hsp90 complexes isolated by specific inhibitors. Such information will lead to better understanding of tumor specific molecular markers to aid in their clinical development. It will also help to elucidate the molecular basis for the biological differences observed among Hsp90 inhibitors.     

逆转录套式PCR检测粪便样本中的SARS冠状病毒

为了观察SARS冠状病毒在SARS患者粪便中的存在规律,建立了检测SARS冠状病毒RNA的逆转录-聚合酶链反应(RT-PCR)方法,并应用该方法检测了241份SARS患者粪便样本。部分PCR产物应用测序技术进行验证。RT-PCR的灵敏度为10^-10稀释度的病毒原液(原液为10^8TCID50／ml)。241份粪便样本的总体检出率为24．1％(58／241),其中发病后的前10d和20d的检出率均为50．0％。随着发病时间的延长,阳性检出率呈下降趋势。应用RT-PCR从粪便中检测SARS冠状病毒是可行的,在发病50d以后仍有17．0％左右的阳性检出率,提示SARS恢复期患者具有排毒的可能性,给后续的卫生防疫措施提供了一定的参考数据。     

miR‐495 sensitizes MDR cancer cells to the combination of doxorubicin and taxol by inhibiting MDR1 expression

MDR1 is highly expressed in MDR A2780DX5 ovarian cancer cells, MDR SGC7901R gastric cancer cells and recurrent tumours. It pumps cytoplasmic agents out of cells, leading to decreased drug accumulation in cells and making cancer cells susceptible to multidrug resistance. Here, we identified that miR‐495 was predicted to target ABCB1, which encodes protein MDR1. To reduce the drug efflux and reverse MDR in cancer cells, we overexpressed a miR‐495 mimic in SGC7901R and A2780DX cells and in transplanted MDR ovarian tumours in vivo. The results indicated that the expression of MDR1 in the above cells or tumours was suppressed and that subsequently the drug accumulation in the MDR cells was decreased, cell death was increased, and tumour growth was inhibited after treatment with taxol‐doxorubicin, demonstrating increased drug sensitivity. This study suggests that pre‐treatment with miR‐495 before chemotherapy could improve the curative effect on MDR1‐based MDR cancer.