B6-Chr1~(LY)和B6-Chr1~(SJ)小鼠1号染色体遗传变异信息的解析

[目的]鉴定B6-Chr1LY和B6-Chr1SJ小鼠品系1号染色体上的遗传变异。[方法]采用Illumina测序平台获得B6-Chr1LY和B6-Chr1SJ品系的全基因组序列并比对到参考基因组。利用Samtools/Bcftools以及speedseq软件鉴定单核苷酸多态性(single nucleotide polymorphism,SNP)、插入缺失(indel)和结构变异,并对其进行功能注释。[结果]两个品系共鉴定380万个SNPs、55万个indels、10 123个大片段缺失、152个重复和87个倒位。二者共享1 642 629 SNPs和190 248个indels;近16.1%SNPs和20.7%indels未在小鼠基因组计划和db SNP142数据集中报道。大片段缺失长度在51~70 kb之间,2 kb的占总数的76%。功能注释发现上百个蛋白编码基因含有非同义突变、移码突变、编码序列缺失等功能性变异。[结论]B6-Chr1LY和B6-Chr1SJ小鼠1号染色体上含有丰富的遗传变异信息,可为后续利用这两个品系的遗传学研究奠定基础。     

Agonist-induced down-regulation of α_(1B)-adrenergic receptor in HEK293 cells transfected with α_(1B)cDNA

α1ａｄｒｅｎｅｒｇｉｃｒｅｃｅｐｔｏｒｓ(α1ＡＲ)ｂｅｌｏｎｇｔｏｔｈｅｓｕｐｅｒｆａｍｉｌｙｏｆＧｐｒｏｔｅｉｎｃｏｕｐｌｅｄｒｅｃｅｐｔｏｒｓ.Ｐｒｏｌｏｎｇｅｄａｃｔｉｖａｔｉｏｎｏｆｔｈｅｓｅｒｅｃｅｐｔｏｒｓｗｏｕｌｄｌｅａｄｔｏａｄｅｃｒｅａｓｅｉｎａｇｏｎｉｓｔｓｅｎｓｉｔｉｖｉｔｙ.Ｔｈｅｐｒｏｃｅｓｓｍａｙｂｅｒｅｌａｔｅｄｔｏｒｅｃｅｐｔｏｒｄｏｗｎｒｅｇｕｌａｔｉｏｎ,ｓｅｑｕｅｓｔｒａｔｉｏｎａｎｄｐｈｏｓｐｈｏｌａｔｉｏｎ,ｅｔｃ.Ｄｅｓｅｎｓｉｔｉｚａｔｉｏｎｏｆα1ＢＡＲ…     

第11届国际生物奥林匹克竞赛(2000)理论试题B(1)

细胞生物学1.删除2 .下列陈述是关于通过细胞膜的转运。在下列空格中从左向右以逐渐增大的顺序 1)～ 7)写出正确陈述的序号。1)大分子被胞吞作用摄入细胞内 ;2 )离子和糖被胞吞作用摄入细胞内 ;3)胶体溶液 (蛋白质、脂类、碳水化合物 )被胞饮作用摄入细胞内 ;4 )脂类可溶性化合物被通道蛋白摄入细胞内 ;5 )细菌被吞噬作用摄入细胞内 ;6)细胞废物被吞噬作用摄入细胞内 ;7)离子通过通道蛋白3.删除4 .在下面 ,你能看到一些蛋白质 ,并列举了一些蛋白质的功能 [(1)～ 8) ]。在蛋白质前方空白处写上适合的功能的序号 (一种蛋白质可能有多于一种的…     

阿扎霉素B(Azalomycin B)研究进展

介绍阿扎霉素B的生物学来源,研究历史,主要理化性质和结构确定,同时阐述了阿扎霉素B的生物合成途径,构效关系,生物学特性和应用前景。     

罗勒籽脂肪酸及其对蛋白酪氨酸磷酸酯酶1B(PTP1B)抑制作用的研究

利用索氏提取法提取罗勒籽油,向罗勒籽油加入氢氧化钾甲醇溶液后并用水浴加热,加入正已烷和蒸馏水萃取,上清液即为罗勒籽油中脂肪酸,用气相色谱质谱法(CC/MS)对脂肪酸进行鉴定.共鉴定出了4种脂肪酸,其中α-亚麻酸为62.88％、亚油酸为20.66％、棕榈酸为10.67％、硬脂酸为5.79％.对罗勒籽脂肪酸进行PTP1B的抑制作用研究,结果表明脂肪酸对PTP1B有较强的抑制作用,其IC50为11.12 μg/mL.该研究为深入研究罗勒籽的药理作用提供了科学依据.     

Liver Kinase B1 Suppresses Lipopolysaccharide-induced Nuclear Factor κB (NF-κB) Activation in Macrophages

Liver kinase B1 (LKB1), a serine/threonine kinase, is a tumor suppressor and metabolic regulator. Recent data suggest that LKB1 is essential in regulating homeostasis of hematopoietic cells and immune responses. However, its role in macrophages and innate immune system remains unclear. Here we report that macrophage LKB1 inhibits pro-inflammatory signaling in response to LPS. LPS-induced pro-inflammatory cytokines and pro-inflammatory enzymes were monitored in bone marrow-derived macrophages isolated from myeloid cell-specific LKB1 knock out mice and their wild type littermate control mice. LPS induced higher levels of pro-inflammatory cytokines and pro-inflammatory enzymes in bone marrow-derived macrophages from LKB1 KO than those from wild type mice. Consistently, LPS induced higher levels of NF-κB activation in LKB1-deficient macrophages than those in wild type. Further, LPS stimulation significantly increased LKB1 phosphorylation at serine 428, which promoted its binding to IκB kinaseβ (IKKβ), resulting in the inhibition of NF-κB. Finally, LPS injection caused higher levels of cytokine release and more severe tissue injury in the lung tissues of LKB1 KO mice than in those of control mice. We conclude that LKB1 inhibits LPS-induced NF-κB activation in macrophages.     

Protein-tyrosine phosphatase 1B (PTP1B) deficiency confers resistance to transforming growth factor-β (TGF-β)-induced suppressor effects in hepatocytes

Transforming growth factor-β (TGF-β) plays a dual role in hepatocytes, mediating both tumor suppressor and promoter effects. The suppressor effects of the cytokine can be negatively regulated by activation of survival signals, mostly dependent on tyrosine kinase activity. The aim of our work was to study the role of the protein-tyrosine phosphatase 1B (PTP1B) on the cellular responses to TGF-β, using for this purpose immortalized neonatal hepatocytes isolated from both PTP1B(+/+) and PTP1B(-/-) mice. We have found that PTP1B deficiency conferred resistance to TGF-β suppressor effects, such as apoptosis and growth inhibition, correlating with lower Smad2/Smad3 activation. Both responses were recovered in the presence of the general tyrosine kinase inhibitor genistein. PTP1B(-/-) cells showed elevated NF-κB activation in response to TGF-β. Knockdown of the NF-κB p65 subunit increased cell response in terms of Smads phosphorylation and apoptosis. Interestingly, these effects were accompanied by inhibition of Smad7 up-regulation. In addition, lack of PTP1B promoted an altered NADPH oxidase (NOX) expression pattern in response to TGF-β, strongly increasing the NOX1/NOX4 ratio, which was reverted by genistein and p65 knockdown. Importantly, NOX1 knockdown inhibited nuclear translocation of p65, promoted Smad phosphorylation, and decreased Smad7 levels. In summary, our results suggest that PTP1B deficiency confers resistance to TGF-β through Smad inhibition, an effect that is mediated by NOX1-dependent NF-κB activation, which in turn, increases the level of the Smad inhibitor Smad7 and participates in a positive feedback loop on NOX1 up-regulation.     

Lysosomal-associated Transmembrane Protein 4B (LAPTM4B) Decreases Transforming Growth Factor β1 (TGF-β1) Production in Human Regulatory T Cells

Production of active TGF-β1 is one mechanism by which human regulatory T cells (Tregs) suppress immune responses. This production is regulated by glycoprotein A repetitions predominant (GARP), a transmembrane protein present on stimulated Tregs but not on other T lymphocytes (Th and CTLs). GARP forms disulfide bonds with proTGF-β1, favors its cleavage into latent inactive TGF-β1, induces the secretion and surface presentation of GARP·latent TGF-β1 complexes, and is required for activation of the cytokine in Tregs. We explored whether additional Treg-specific protein(s) associated with GARP·TGF-β1 complexes regulate TGF-β1 production in Tregs. We searched for such proteins by yeast two-hybrid assay, using GARP as a bait to screen a human Treg cDNA library. We identified lysosomal-associated transmembrane protein 4B (LAPTM4B), which interacts with GARP in mammalian cells and is expressed at higher levels in Tregs than in Th cells. LAPTM4B decreases cleavage of proTGF-β1, secretion of soluble latent TGF-β1, and surface presentation of GARP·TGF-β1 complexes by Tregs but does not contribute to TGF-β1 activation. Therefore, LAPTM4B binds to GARP and is a negative regulator of TGF-β1 production in human Tregs. It may play a role in the control of immune responses by decreasing Treg immunosuppression.     

问题解答(1)

问:绿色植物在有阳光时行光合作用吸二氧化碳放出氧气,在晚上没阳光时只有呼吸作用吸氧气放二氧化碳,为什么总说早晨到树林中散步空气好呢? 答:因为空气流动得很厉害,所以在一般树林中白天夜晚O_2及CO_2含量的改变是极小的.说早晨空气好,并不是植物起了什么作用,而是因为夜间人们的活动停止了,车马不在路上奔驰了,工厂的烟囱不冒烟了.加上树林中的湿度较大,尘土都降落在地上,空气就更为清洁了.我们夜间睡在屋中,屋中温度较高,不好气味的有机物(器物中发出或人呼出)空气中也很多,所以一到树林中,凉爽而清洁的空气,眼界的开朗就使我们心神为之一振.(吴相钰、董愚得答)     

B.B.Brodie(1907～1989)

B.B.Brodie博土,一个先驱药理学家和美国国家科学院院士,1989年2月28日逝世。生前是美国国立卫生研究院心脏研究所化学药理实验室的创始人和第一任主任。     

Palmitate-induced PTP1B expression is mediated by ceramide-JNK and nuclear factor κB (NF-κB) activation

Palmitate induces PTP1B expression in skeletal muscle cells. The purpose of this study was to investigate the mechanisms responsible for palmitate-induced PTP1B expression in mouse skeletal muscle cells. Three truncated fragments of PTP1B promoter were cloned into PGL3-basic vector and the promoter activity of PTP1B was assessed in C2C12 cells exposed to palmitate either in the presence or in the absence of several inhibitors to study the biochemical pathways involved. EMSA was performed to examine binding of NF-κB to NF-κB consensus sequence and PTP1B oligonucelotides in the cells treated with palmitate. Lentiviral PTP1B-shRNA was used to knockdown PTP1B in myotubes. The phosphorylation and protein levels of IRS-1 and Akt were detected by western blot. 0.5mM palmitate induced PTP1B promoter activity in fragment -1715/+59 by 50% (p<0.01). Palmitate increased NF-κB binding to both NF-κB consensus sequence and one NF-κB sequence (-920 to -935) in PTP1B promoter. Incubation of C2C12 cells with different concentrations of C2-ceramide enhanced PTP1B promoter activity dose-dependently. Inhibitors of de novo ceramide synthesis prevented palmitate-induced PTP1B promoter activity in myotubes. In addition, inhibitor of JNK pathway prevented ceramide-induced PTP1B promoter activity in myotubes. Knockdown of PTP1B also prevented ceramide-reduced IRS-1 and Akt phosphorylations in the myotubes. Exposure of the cells to PMA and calphostin C, an inhibitor of PKC, did not affect the activity of PTP1B promoter. Our data provide the evidence that the mechanism by which palmitate increased the expression of PTP1B seems to be through a mechanism involving the activation of ceramide-JNK and NF-κB pathways.     

One-step partial synthesis of (±)-asperteretone B and related hPTP1B1–400 inhibitors from butyrolactone I

Protein tyrosine phosphatase 1B (PTP1B) is a validated target for developing antiobesity, antidiabetic and anticancer drugs. Over the past years, several inhibitors of PTP1B have been discovered; however, none has been approved by the drug regulatory agencies. Interestingly, the research programs focused on discovering PTP1B inhibitors typically use truncated structures of the protein (PTP1B_1-300, 1–300 amino acids), leading to the loss of valuable information about the inhibition and selectivity of ligands and repeatedly misleading the optimization of putative drug leads. Up to date, only six inhibitors of the full-length protein (hPTP1B_1-400), with affinity constants ranging from 1.3 × 10⁴ to 3.3 × 10⁶ M⁻¹, have been reported. Towards the discovery of new ligands of the full-length human PTP1B (hPTP1B_1-400) from natural sources, herein we describe the isolation of a γ-lactone (1, butyrolactone I) from the fungus Aspergillus terreus, as well as the semisynthesis, inhibitory properties (in vitro and in silico), and the structure-activity relationship of a set of butyrolactone derivatives (1 and 2, and 6–12) as hPTP1B_1-400 inhibitors, as well as the affinity constant (k_a = 2.2 × 10⁵ M⁻¹) of the 1-hPTP1B₁–₄₀₀ complex, which was determined by fluorescence quenching experiments, after the inner filter effect correction.     

B链N端去二肽(B1～2)C端去五肽(B26～30)胰岛素晶体结构的分子置换法研究

以B链羧端去五肽胰岛素(DPI)结构为模型,应用分子置换法对B链N端去二肽(B1～2)C端去五肽(B26～30)胰岛素(DesB1～2DPI)晶体结构进行了研究.DesB1～2DPI晶体单位晶胞中每个结晶学不对称单位包含1个DesB1～2DPI分子.交叉旋转函数和平移函数搜索均找到了明显突出的峰,确定了DesB1～2DPI分子在晶胞中的取向和位置.进一步的三维模型重建和结构精化结果巩固了DesB1～2DPI的分子置换法研究的正确性.     

Bücherbesprechung (Book Reviews)

Abstract

A field experiment was conducted during 2002 – 2003 and 2003 – 2004 to study the impact of controlling the insect vector, Whitefly, of yellow mosaic virus on the performance of green gram grown under rainfed lowland rice fallow situation. Insecticide, Imidachloprid 12.5 kg a.i./ha, was applied at 15- and 30-days growth to four green gram varieties, direct-sown under residual soil moisture conditions after the harvest of kharif rice. Results showed crops under YMV-management achieved 65.67% more seed yield producing 11.68 q/ha, which was significantly higher than the crops under no YMV management (7.05 q ha^?1). Plants which had been treated with insecticide were found to be less affected (8.50%) by YMV compared to those which had no treatment (20.10%). Varieties PDM 54 and PDM 11 emerged as superior, producing significantly higher seed yields, 12.73 and 11.69 q/ha respectively; while Pusa vishal (8.21 q ha^?1) and Pusa 105 (7.85 q ha^?1) produced moderately compared with the local variety (6.31 q ha^?1). In addition, PDM 54 and PDM 11 showed relatively more resistance against YMV infestation recording 5.23% and 6.01% infected plants.     

Regulation of α(2B)-adrenergic receptor-mediated extracellular signal-regulated kinase 1/2 (ERK1/2) activation by ADP-ribosylation factor 1

A number of signaling molecules are involved in the activation of the mitogen-activated protein kinase (MAPK) pathway by G protein-coupled receptors. In this study, we have demonstrated that α(2B)-adrenergic receptor (α(2B)-AR) interacts with ADP-ribosylation factor 1 (ARF1), a small GTPase involved in vesicle-mediated trafficking, in an agonist activation-dependent manner and that the interaction is mediated through a unique double Trp motif in the third intracellular loop of the receptor. Interestingly, mutation of the double Trp motif and siRNA-mediated depletion of ARF1 attenuate α(2B)-AR-mediated activation of extracellular signal-regulated kinases 1/2 (ERK1/2) without altering receptor intracellular trafficking, whereas expression of the constitutively active mutant ARF1Q71L and ARNO, a GDP-GTP exchange factor of ARF1, markedly enhances the activation of Raf1, MEK1, and ERK1/2. These data strongly demonstrate that the small GTPase ARF1 modulates ERK1/2 activation by α(2B)-AR and provide the first evidence indicating a novel function for ARF1 in regulating the MAPK signaling pathway.     

恐龙遗址旅游指南(3)欧洲篇(中B)

第三站:英国西德纳姆镇(Sydenhm)水晶宫公园(Crystal Palace Park) 离开刘易斯镇,我们原路返回伦敦市,继续这次朝圣之旅.我们先到著名的伦敦桥南端的伦敦桥站火车站(London Bridge),这个车站于1836年12月开始运营,以连结东南郊的通勤及短程列车为主,因此车站的早晚高峰时间极为拥挤混杂.     

Synergistic Suppression of Early Phase of Adipogenesis by Microsomal PGE Synthase-1 (PTGES1)-Produced PGE(2) and Aldo-Keto Reductase 1B3-Produced PGF(2α)

We recently reported that aldo-keto reductase 1B3-produced prostaglandin (PG) F(2α) suppressed the early phase of adipogenesis. PGE(2) is also known to suppress adipogenesis. In this study, we found that microsomal PGE(2) synthase (PGES)-1 (mPGES-1; PTGES1) acted as the PGES in adipocytes and that PGE(2) and PGF(2α) synergistically suppressed the early phase of adipogenesis. PGE(2) production was detected in preadipocytes and transiently enhanced at 3 h after the initiation of adipogenesis of mouse adipocytic 3T3-L1 cells, followed by a quick decrease; and its production profile was similar to the expression of the cyclooxygenase-2 (PTGS2) gene. When 3T3-L1 cells were transfected with siRNAs for any one of the three major PTGESs, i.e., PTGES1, PTGES2 (mPGES-2), and PTGES3 (cytosolic PGES), only PTGES1 siRNA suppressed PGE(2) production and enhanced the expression of adipogenic genes. AE1-329, a PTGER4 (EP4) receptor agonist, increased the expression of the Ptgs2 gene with a peak at 1 h after the initiation of adipogenesis. PGE(2)-mediated enhancement of the PTGS2 expression was suppressed by the co-treatment with L-161982, a PTGER4 receptor antagonist. Moreover, AE1-329 enhanced the expression of the Ptgs2 gene by binding of the cyclic AMP response element (CRE)-binding protein to the CRE of the Ptgs2 promoter; and its binding was suppressed by co-treatment with L-161982, which was demonstrated by promoter luciferase and chromatin immunoprecipitation assays. Furthermore, when 3T3-L1 cells were caused to differentiate into adipocytes in medium containing both PGE(2) and PGF(2α), the expression of the adipogenic genes and the intracellular triglyceride level were decreased to a greater extent than in medium containing either of them, revealing that PGE(2) and PGF(2α) independently suppressed adipogenesis. These results indicate that PGE(2) was synthesized by PTGES1 in adipocytes and synergistically suppressed the early phase of adipogenesis of 3T3-L1 cells in cooperation with PGF(2α) through receptor-mediated activation of PTGS2 expression.