BACKGROUND & AIMS: :As the malaria parasite, Plasmodium falciparum, grows within its host erythrocyte it induces an increase in the permeability of the erythrocyte membrane to a range of low-molecular-mass solutes, including Na+ and K+ (ref. 1). This results in a progressive increase in the concentration of Na+ in the erythrocyte cytosol. The parasite cytosol has a relatively low Na+ concentration and there is therefore a large inward Na+ gradient across the parasite plasma membrane. Here we show that the parasite exploits the Na+ electrochemical gradient to energize the uptake of inorganic phosphate (P(i)), an essential nutrient. P(i) was taken up into the intracellular parasite by a Na+-dependent transporter, with a stoichiometry of 2Na+:1P(i) and with an apparent preference for the monovalent over the divalent form of P(i). A P(i) transporter (PfPiT) belonging to the PiT family was cloned from the parasite and localized to the parasite surface. Expression of PfPiT in Xenopus oocytes resulted in Na+-dependent P(i) uptake with characteristics similar to those observed for P(i) uptake in the parasite. This study provides new insight into the significance of the malaria-parasite-induced alteration of the ionic composition of its host cell.

背景与目标： : 由于疟原虫恶性疟原虫在其宿主红细胞内生长，因此引起红细胞膜对一系列低分子质量溶质 (包括Na和K) 的渗透性增加 (参考文献1)。这会导致红细胞胞质溶胶中Na的浓度逐渐增加。寄生虫胞质溶胶的Na浓度相对较低，因此在整个寄生虫质膜上存在较大的向内Na梯度。在这里，我们表明寄生虫利用Na电化学梯度来激发无机磷酸盐 (P(i)) 的吸收，这是一种必需的营养素。P(i) 被Na依赖性转运蛋白吸收到细胞内寄生虫中，化学计量为2Na: 1P(i)，并且明显偏爱单价而不是P(i) 的二价形式。从寄生虫中克隆了属于PiT家族的P(i) 转运蛋白 (PfPiT)，并定位在寄生虫表面。爪蟾卵母细胞中PfPiT的表达导致Na依赖性P(i) 摄取，其特征与在寄生虫中观察到的P(i) 摄取相似。这项研究为疟疾寄生虫诱导的宿主细胞离子组成改变的意义提供了新的见解。

BACKGROUND & AIMS: OBJECTIVE:Evaluate in patients with celiac disease the tolerance of prolonged consumption of small amounts of gliadin contained in products containing wheat starch.

DESIGN:Open 1-year trial of the addition of wheat starch to a gluten-free diet in a cohort of adult patients with biopsy-proven celiac disease who had never consumed wheat starch. The control group consisted of patients with celiac disease who tolerated wheat starch.

SUBJECTS:Seventeen patients with celiac disease and 14 control patients, all diagnosed according to criteria of the European Society of Pediatric Gastroenterology and Nutrition, were recruited from the Canadian Celiac Association and the Quebec Celiac Foundation.

SETTING:The study was conducted in the outpatient clinic of the Gastroenterology and Nutrition Service of Ste Justine Hospital, Montreal, Quebec, Canada.

INTERVENTIONS:Patients were asked to consume four to six portions daily of a wheat starch-containing product, mainly bread, for up to 1 year.

MAIN OUTCOME MEASURES:The gliadin content of the wheat starch product used in this trial was quantified by enzyme-linked immunosorbent assay. Patient outcome measures included symptoms, nutritional parameters (anthropometric data, complete blood count, serum folate and iron levels), and immunologic parameters (antigliadin antibody and antiendomysium antibody titers).

RESULTS:A quantifiable amount of immunoreactive gliadin (0.75 mg/100 g) was found in the wheat starch. The majority of the patients with celiac disease (11 of 17) who had never consumed wheat starch previously developed symptoms, which resolved within weeks of discontinuing the product. Relapse of skin lesions was seen in two of three patients with coexisting dermatitis herpetiformis. No weight loss or biochemical changes were observed. Despite the presence of symptoms, antigliadin antibody and antiendomysium antibody determinations were not useful to detect the clinical intolerance.

APPLICATIONS:The innocuousness of the long-term ingestion of "gluten-free" products containing wheat starch is still unproven, and prolonged use of such products by patients with celiac disease cannot be recommended.

背景与目标： 目标 : 评估乳糜泻患者对含有小麦淀粉的产品中含有的少量麦醇溶蛋白的长期食用的耐受性。
设计 : 在从未食用过小麦淀粉的活检证实的乳糜泻成年患者队列中，将小麦淀粉添加到无麸质饮食中进行了为期1年的开放试验。对照组由耐受小麦淀粉的乳糜泻患者组成。
受试者 : 17例乳糜泻患者和14例对照患者，均根据欧洲儿科胃肠病与营养学会的标准进行诊断，是从加拿大乳糜泻协会和魁北克乳糜泻基金会招募的。
设置 : 该研究是在魁北克蒙特利尔Ste Justine医院胃肠病学和营养服务门诊进行的，加拿大。
干预措施 : 要求患者每天食用四到六份含小麦淀粉的产品，主要是面包，长达1年。
主要结果指标 : 通过酶联免疫吸附测定法对该试验中使用的小麦淀粉产品的麦醇溶蛋白含量进行了定量。患者结局指标包括症状，营养参数 (人体测量数据，全血细胞计数，血清叶酸和铁水平) 和免疫学参数 (抗麦醇溶蛋白抗体和抗肌内膜抗体滴度)。
结果 : 在小麦淀粉中发现可定量的免疫反应性麦醇溶蛋白 (0.75 mg/100g)。以前从未食用过小麦淀粉的大多数乳糜泻患者 (17人中有11人) 出现症状，这些症状在停用该产品后的几周内得到缓解。在三名并存的疱疹样皮炎患者中，有两名发现了皮肤病变的复发。未观察到体重减轻或生化变化。尽管存在症状，但抗麦醇溶蛋白抗体和抗肌内膜抗体的测定对检测临床不耐受没有用。
应用 : 长期摄入含有小麦淀粉的 “无麸质” 产品的无伤害性仍未得到证实，并且不建议乳糜泻患者长期使用此类产品。

BACKGROUND & AIMS: The stability and activity of indole-3-glycerol phosphate synthase from Sulfolobus solfataricus were studied as a function of pH and temperature. In this paper we focus on three points(1) the long-term stability of the protein to irreversible denaturation at high temperature; (2) the short-term stability of the protein to reversible temperature-driven unfolding; and (3) the dependence of its activity on temperature.

Results can be summarized as follows(a) the same first-order kinetic constant (0.020+/-0.003 min-1) was determined at different pH values (6.5, 8.0 and 9.5) from long-term stability experiments at 80 degrees C; (b) short-term stability experiments revealed different behaviour in two different pH ranges (6.5-8.0, 8.5-9.5), suggesting that the melting temperature is higher at alkaline than at neutral pH; (c) the dependence of activity on temperature was investigated at pH 7.0 and 9.0, and a discontinuity was observed in the Arrhenius plot of kcat values at pH 9.0. We also investigated the stability in the presence of guanidinium chloride at 20 degrees C either at pH 7.0 or at pH 9.0, and we present data that indicate that the unfolding mechanism closely approaches a two-state model at pH 7.0 and a more complex mechanism at pH 9.0. Satisfactory fitting of the equilibrium unfolding transition obtained by fluorescence measurements at pH 9.0 required a model that involves a stable intermediate in addition to the native and unfolded forms. At 20 degrees C the folded conformation is more stable than the unfolded conformation by (14. 7+/-1.2) kJ/mol at pH 7.0 and by (25.5+/-1.8) kJ/mol at pH 9.0.

背景与目标： 研究了硫酸亚砜中吲哚-3-甘油磷酸合酶的稳定性和活性随pH和温度的变化。在本文中，我们重点关注三点 (1) 蛋白质在高温下不可逆变性的长期稳定性; (2) 蛋白质在可逆温度驱动的去折叠的短期稳定性; (3) 其活性对温度的依赖性。
结果可以总结如下 (a) 在不同的ph值 (6.5，8.0和9.5) 来自80 ℃ 下的长期稳定性实验; (b) 短期稳定性实验揭示了在两个不同pH范围 (6.5-8.0，8.5-9.5) 下的不同行为，表明在碱性下的熔融温度高于在中性pH下; (c) 在pH 7.0和9.0下研究了活性对温度的依赖性，在pH 9.0下的kcat值的Arrhenius图中观察到不连续性。我们还研究了氯化胍在20 ℃ 下在pH 7.0或pH 9.0下的稳定性，并且我们提供的数据表明，展开机制在pH 7.0下接近两态模型，在pH 9.0下接近更复杂的机制。在pH 9.0下通过荧光测量获得的平衡展开转变的令人满意的拟合需要一个模型，该模型除了天然和展开形式之外还涉及稳定的中间体。在20 ℃ 下，折叠构象比未折叠构象在pH 7.0下稳定 (14. 7 +/-1.2) kJ/mol，在pH 9.0下稳定 (25.5 +/-1.8) kJ/mol。

BACKGROUND & AIMS: :The anaerobic metabolism of phenol proceeds via carboxylation to 4-hydroxybenzoate by a two-step process involving seven proteins and two enzymes ("biological Kolbe-Schmitt carboxylation"). MgATP-dependent phosphorylation of phenol catalyzed by phenylphosphate synthase is followed by phenylphosphate carboxylation. Phenylphosphate synthase shows similarities to phosphoenolpyruvate (PEP) synthase and was studied for the bacterium Thauera aromatica. It consists of three proteins and transfers the beta-phosphoryl from ATP to phenol; the products are phenylphosphate, AMP, and phosphate. We showed that protein 1 becomes phosphorylated in the course of the reaction cycle by [beta-(32)P]ATP. This reaction requires protein 2 and is severalfold stimulated by protein 3. Stimulation of the reaction by 1 M sucrose is probably due to stabilization of the protein(s). Phosphorylated protein 1 transfers the phosphoryl group to phenolic substrates. The primary structure of protein 1 was analyzed by nanoelectrospray mass spectrometry after CNBr cleavage, trypsin digestion, and online high-pressure liquid chromatography at alkaline pH. His-569 was identified as the phosphorylated amino acid. We propose a catalytic ping-pong mechanism similar to that of PEP synthase. First, a diphosphoryl group is transferred to His-569 in protein 1, from which phosphate is cleaved to render the reaction unidirectional. Histidine phosphate subsequently serves as the actual phosphorylation agent.

背景与目标： : 苯酚的厌氧代谢通过涉及七种蛋白质和两种酶的两步过程 (“生物Kolbe-Schmitt羧化”) 通过羧化为4-羟基苯甲酸酯。由苯基磷酸合酶催化的苯酚的MgATP依赖性磷酸化，然后是苯基磷酸羧化。苯基磷酸合酶与磷酸烯醇丙酮酸 (PEP) 合酶相似，并对细菌Thauera aromatica进行了研究。它由三种蛋白质组成，并将 β-磷酰基从ATP转移到苯酚; 产品是磷酸苯基，AMP和磷酸盐。我们证明了蛋白质1在反应循环过程中被 [β-(32)P]ATP磷酸化。该反应需要蛋白质2，并且被蛋白质3刺激了几倍。1 m蔗糖对反应的刺激可能是由于蛋白质的稳定。磷酸化蛋白质1将磷酰基转移到酚类底物上，CNBr裂解后通过纳米电喷雾质谱分析蛋白质1的一级结构，胰蛋白酶消化和在线高压液相色谱在碱性pH下。His-569被鉴定为磷酸化氨基酸。我们提出了类似于PEP合酶的催化乒乓机制。首先，在蛋白质1中，二磷酰基转移至His-569，从中裂解磷酸盐以使反应单向。磷酸组氨酸随后用作实际的磷酸化剂。

BACKGROUND & AIMS: :Methyl-β-cyclodextrin (MβCD) reduces lysosomal cholesterol accumulation in Niemann-Pick disease type C1 (NPC1) patient fibroblasts. However, the pharmacological activity of MβCD reported by different laboratories varies. To determine the potential causes of this variation, we analyzed the mass spectrum characteristics, pharmacological activity of three preparations of MβCDs, and the protein expression profiles of NPC1 patient fibroblasts after treatment with different sources of MβCDs. Our data revealed varied mass spectrum profiles and pharmacological activities on the reduction of lysosomal cholesterol accumulation in NPC1 fibroblasts for these three preparations of MβCDs obtained from different batches and different sources. Furthermore, a proteomic analysis showed the differences of these three MβCD preparations on amelioration of dysregulated protein expression levels in NPC1 cells. The results demonstrate the importance of prescreening of different cyclodextrin preparations before use as a therapeutic agent. A combination of mass spectrum analysis, measurement of pharmacological activity, and proteomic profiling provides an effective analytical procedure for characterization of cyclodextrins for therapeutic applications.

背景与目标： : 甲基-β-环糊精 (m β cd) 减少Niemann-Pick病C1 (NPC1) 患者成纤维细胞中的溶酶体胆固醇积累。但是，不同实验室报道的m β cd的药理活性有所不同。为了确定这种变异的潜在原因，我们分析了三种m β cds制剂的质谱特征，药理活性以及用不同来源的m β cds处理后NPC1患者成纤维细胞的蛋白表达谱。我们的数据揭示了从不同批次和不同来源获得的这三种m β cds制剂在NPC1成纤维细胞中减少溶酶体胆固醇积累的不同质谱谱和药理活性。此外，蛋白质组学分析表明，这三种m β cd制剂在改善NPC1细胞中蛋白表达水平失调方面存在差异。结果表明，在用作治疗剂之前，对不同的环糊精制剂进行预检查的重要性。质谱分析，药理活性测量和蛋白质组学分析相结合，为表征用于治疗应用的环糊精提供了有效的分析程序。

BACKGROUND & AIMS: BACKGROUND AND PURPOSE:Acyl derivatives of CoA have been shown to act as antagonists at human platelet and recombinant P2Y1 receptors, but little is known about their effects in the cardiovascular system. This study evaluated the effect of these endogenous nucleotide derivatives at P2Y1 receptors natively expressed in rat and porcine blood vessels. EXPERIMENTAL APPROACH:Isometric tension recordings were used to evaluate the effects of CoA, acetyl CoA, palmitoyl CoA (PaCoA) and 3'-dephospho-palmitoyl-CoA on concentration relaxation-response curves to ADP and uridine triphosphate (UTP). A FlexStation monitored ADP- and UTP-evoked calcium responses in HEK293 cells. KEY RESULTS:Acetyl CoA and PaCoA, but not CoA, inhibited endothelium-dependent relaxations to ADP with apparent selectivity for P2Y1 receptors (over P2Y(2/4) receptors) in rat thoracic aorta; PaCoA was more potent than acetyl CoA (331-fold vs. fivefold shift of ADP response curve evoked by 10 μM PaCoA and acetyl CoA, respectively); the apparent pA2 value for PaCoA was 6.44. 3'-dephospho-palmitoyl-CoA (10 μM) was significantly less potent than PaCoA (20-fold shift). In porcine mesenteric arteries, PaCoA and the P2Y1 receptor antagonist MRS2500 blocked ADP-mediated endothelium-dependent relaxations; in contrast, they were ineffective against ADP-mediated endothelium-independent relaxation in porcine coronary arteries (which does not involve P2Y1 receptors). Calcium responses evoked by ADP activation of endogenous P2Y1 receptors in HEK293 cells were inhibited in the presence of PaCoA, which failed to alter responses to UTP (acting at endogenous P2Y(2/4) receptors). CONCLUSIONS AND IMPLICATIONS:Acyl derivatives of CoA can act as endogenous selective antagonists of P2Y1 receptors in blood vessels, and this inhibitory effect critically depends on the palmitate and 3'-ribose phosphate substituents on CoA.

背景与目标：

BACKGROUND & AIMS: BACKGROUND:Interleukin-17 (IL-17), which is secreted by Th17 cells, is a proinflammatory cytokine that is implicated in the pathogenesis of multiple sclerosis (MS) and plays a role in nonresponse of MS patients to interferon-β (IFN-β) therapy. OBJECTIVES:The purpose of this study was to establish a correlation between nonresponders (NR) and IL-17A serum titers and binding antibodies (BAbs) to IFN-β, as well as to find a correlation between IL-17A serum levels and other features of MS patients. METHODS:Our prospective study included 72 inactive relapsing-remitting multiple sclerosis (RRMS) patients that had been treated for at least 18 months with IFN-β and 15 healthy subjects. We determined the serum levels of IL-17A and of BAbs. IL-17A levels were considered elevated (IL-17A+) if the recorded value was greater than 1.6 pg/ml. RESULTS:Twenty-seven patients (37.5%) were NR and had a significantly higher serum IL-17A level compared to the responders group. Nineteen patients (26.4%) were IL-17A+ and had had a significantly higher number of relapses in the previous year and a higher Expanded Disability Status Score. The majority of IL-17A+ patients were NR and had a shorter MS duration. CONCLUSIONS:RRMS patients with high serum IL-17A levels do not respond well to IFN-β therapy and have shorter MS duration compared to patients with low IL-17A levels. This response is not influenced by the presence of BAbs.

背景与目标：

BACKGROUND & AIMS: :Molecular dynamics simulations in explicit water were carried out for two stacks, each composed of six 10-strand antiparallel β-sheets for two peptides corresponding to the diverging turn of two homologous Abl-SH3 domains. The first system, referred to as 10×6×MK contained the DLSFMKGE sequence from the Drosophila, while the second one, referred to as 10×6×KK, contained the human DLSFKKGE sequence. It was found that the 10×6×MK β-sheet stack is stable, but the 10×6×KK β-sheet stack is not. The stability of the 10×6×MK β-sheet stack results from the hydrophobic interactions of the methionine and phenylalanine residues and the leucine residues of the neighboring sheets. The Met, Phe, and Leu hydrophobic units make a hydrophobic core for the stack of β-sheets. During the MD run, the Met, Phe, and Leu residues of the neighboring β-sheets acted as a conformational switch moving the β-sheets so that the Phe residue interacted with the Met residue from the neighboring β-sheet. Replacement of Met by Lys destroys the hydrophobic core, which is the stability factor of the β-sheet stack. For the 10×6×KK system, individual β-sheets were preserved during simulations, but the interactions between the β-sheets were lost. The calculations of a six β-sheet stack confirm the conclusion drawn from our earlier studies of single β-sheet systems that the β-sheets must form stacks to be stabilized. These results suggest that the two conserved basic residues at the diverging turn of SH3 domains could act as gatekeepers to avoid aggregation.

背景与目标： : 在显式水中对两个堆栈进行了分子动力学模拟，每个堆栈由两个肽的六个10链反平行 β-折叠组成，对应于两个同源Abl-SH3域的发散转向。第一个系统 (称为10 × 6 × mk) 包含果蝇的DLSFMKGE序列，而第二个系统 (称为10 × 6 × kk) 包含人类DLSFKKGE序列。发现10 × 6 × mk β-折叠堆叠是稳定的，而10 × 6 × kk β-折叠堆叠则不稳定。10 × 6 × mk β-薄片堆叠的稳定性是由蛋氨酸和苯丙氨酸残基与相邻薄片的亮氨酸残基的疏水相互作用引起的。Met，Phe和Leu疏水单元为 β-折叠堆叠形成疏水核心。在MD运行期间，相邻 β-sheet的Met，Phe和Leu残基充当构象开关，移动 β-sheet，以使Phe残基与相邻 β-sheet的Met残基相互作用。用Lys代替Met会破坏疏水核，这是 β-折叠堆叠的稳定性因子。对于10 × 6 × kk系统，在模拟过程中保留了单个 β-折叠，但是 β-折叠之间的相互作用丢失了。六个 β-sheet堆栈的计算证实了我们先前对单个 β-sheet系统的研究得出的结论，即 β-sheet必须形成堆栈才能稳定。这些结果表明，在SH3结构域发散的转弯处的两个保守的碱性残基可以充当网守以避免聚集。

BACKGROUND & AIMS: :As colorectal cancer remains the second highest cause of cancer-related deaths in much of the industrialised world, identifying novel strategies to prevent colorectal tumour development remains an important challenge. BAG-1 is a multi-functional protein, the expression of which is up-regulated at relatively early stages in colorectal tumorigenesis. Importantly, BAG-1 is thought to enhance colorectal tumour progression through promoting tumour cell survival. Here, we report for the first time a novel role for BAG-1, establishing it as a suppressor of transforming growth factor β (TGF-β1) expression in colorectal tumour cells. Microarray analysis first highlighted the possibility that BAG-1 may regulate TGF-β1 expression, a key cytokine in normal colonic tissue homoeostasis. Q-RT-PCR and ELISA demonstrated TGFB1 mRNA and protein expression to be significantly increased when BAG1 levels were reduced by small interfering RNA; additionally, induction of BAG-1L caused suppression of TGFB1 mRNA in colorectal tumour cells. Using reporter and chromatin immunoprecipitation assays, a direct association of BAG-1 with the TGFB1 gene regulatory region was identified. Immunohistochemistry and Weiser fraction data indicated that the levels of BAG-1 and TGF-β1 are inversely correlated in the normal colonic epithelium in vivo, consistent with a role for BAG-1-mediated repression of TGF-β1 production. In vitro studies showed that the change in TGF-β1 production following manipulation of BAG-1 is functionally relevant; through induction of anchorage-independent growth in TGF-β1-dependent normal rat kidney fibroblasts and regulation of SMAD2 phosphorylation in TGF-β1-sensitive adenoma cells. Taken together, this study identifies the anti-apoptotic protein BAG-1 as a suppressor of the inhibitory growth factor TGF-β1, suggesting that high expression of BAG-1 can impact on a number of the hallmarks of cancer, of potential importance in promoting the early stages of colorectal tumorigenesis. Establishing BAG-1 as a repressor of TGF-β1 has important biological implications, and highlights a new role for BAG-1 in colorectal tumorigenesis.

背景与目标： : 由于大肠癌仍然是许多工业化国家中癌症相关死亡的第二大原因，因此确定预防大肠癌发展的新策略仍然是一项重要挑战。BAG-1是一种多功能蛋白，其表达在结直肠肿瘤发生的相对早期阶段上调。重要的是，BAG-1被认为可以通过促进肿瘤细胞存活来促进结直肠肿瘤的进展。在这里，我们首次报道了BAG-1的新作用，将其确立为大肠肿瘤细胞中转化生长因子 β (TGF-β1) 表达的抑制因子。微阵列分析首先强调了BAG-1可能调节TGF-β1表达的可能性，TGF-β1是正常结肠组织平衡中的关键细胞因子。Q-rt-pcr和ELISA证明，当小干扰RNA降低BAG1水平时，TGFB1 mRNA和蛋白表达显着增加; 此外，BAG-1L的诱导引起大肠肿瘤细胞中TGFB1 mRNA的抑制。使用报告基因和染色质免疫沉淀测定法，鉴定了BAG-1与TGFB1基因调控区的直接关联。免疫组织化学和Weiser分数数据表明，体内正常结肠上皮中BAG-1和TGF-β1的水平呈负相关，这与BAG-1介导的TGF-β1产生抑制作用一致。体外研究表明，操纵BAG-1后TGF-β1产生的变化在功能上是相关的; 通过诱导TGF-β1依赖性正常大鼠肾脏成纤维细胞中的锚定依赖性生长以及调节TGF-β1敏感性腺瘤细胞中的SMAD2磷酸化。总之，这项研究确定了抗凋亡蛋白BAG-1作为抑制性生长因子TGF-β1的抑制因子，表明BAG-1的高表达可以影响许多癌症的特征，在促进结直肠肿瘤发生的早期阶段具有潜在的重要性。建立BAG-1作为TGF-β1的阻遏物具有重要的生物学意义，并突出了BAG-1在结直肠肿瘤发生中的新作用。

BACKGROUND & AIMS: :Human coronary artery endothelial cells (HCAECs) have the potential to undergo fibrogenic endothelial-mesenchymal transition (EndMT), which results in matrix-producing fibroblasts and thereby contributes to the pathogenesis of cardiac fibrosis. Recently, the profibrotic cytokine transforming growth factor-β (TGF-β) is shown to be the crucial pathogenic driver which has been verified to induce EndMT. C-Ski is an important regulator of TGF-β signaling. However, the detailed role of c-Ski and the molecular mechanisms by which c-Ski affects TGF-β-induced EndMT in HCAECs are not largely elucidated. In the present study, we treated HCAECs with TGF-β of different concentrations to induce EndMT. We found that overexpression of c-Ski in HCAECs either blocked EndMT via hindering Vimentin, Snail, Slug, and Twist expression while enhancing CD31 expression, with or without TGF-β treatment. In contrast, suppression of c-Ski further enhanced EndMT. Currently, miRNA expression disorder has been frequently reported associating with cardiac fibrosis. By using online tools, we regarded miR-155 as a candidate miRNA that could target c-Ski, which was verified using luciferase assays. C-Ski expression was negatively regulated by miR-155. TGF-β-induced EndMT was inhibited by miR-155 silence; the effect of TGF-β on Vimentin, CD31, Snail, Slug, and Twist could be partially restored by miR-155. Altogether, these findings will shed light on the role and mechanism by which miR-155 regulates TGF-β-induced HCAECs EndMT via c-Ski to affect cardiac fibrosis, and miR-155/c-Ski may represent novel biomarkers and therapeutic targets in the treatment of cardiac fibrosis.

背景与目标： : 人冠状动脉内皮细胞 (HCAECs) 具有发生纤维化的内皮-间质转化 (EndMT) 的潜力，这导致产生基质的成纤维细胞，从而有助于心脏纤维化的发病机理。最近，纤维化细胞因子转化生长因子-β (TGF-β) 被证明是关键的致病驱动因素，已被证实可诱导EndMT。C-ski是TGF-β 信号的重要调节因子。然而，c-Ski的详细作用以及c-Ski影响HCAECs中TGF-β 诱导的EndMT的分子机制尚未得到很大的阐明。在本研究中，我们用不同浓度的TGF-β 处理HCAECs以诱导EndMT。我们发现，有或没有TGF-β 处理，HCAECs中c-Ski的过表达通过阻碍波形蛋白，蜗牛，Slug和Twist的表达来阻断EndMT，同时增强CD31的表达。相比之下，抑制c-ski进一步增强了EndMT。目前，miRNA表达紊乱经常被报道与心脏纤维化有关。通过使用在线工具，我们将miR-155视为可以靶向c-Ski的候选miRNA，使用荧光素酶分析验证了这一点。C-Ski表达受miR-155负调节。TGF-β 诱导的EndMT被miR-155沉默抑制; TGF-β 对波形蛋白，CD31，蜗牛，Slug和Twist的作用可以通过miR-155部分恢复。总之，这些发现将阐明miR-155通过c-Ski调节TGF-β 诱导的HCAECs EndMT以影响心脏纤维化的作用和机制，并且miR-155/c-Ski可能代表治疗心脏纤维化的新型生物标志物和治疗靶标。

BACKGROUND & AIMS: OBJECTIVE:Marrow-stimulation techniques are used by surgeons to repair cartilage lesions although consistent regeneration of hyaline cartilage is rare. We have shown previously that autologous blood can be mixed with a polymer solution containing chitosan in a glycerol phosphate (GP) buffer (chitosan-GP), and that implantation of this polymer/blood composite onto marrow-stimulated chondral defects in rabbit and sheep leads to the synthesis of more chondral repair tissue with greater hyaline character compared to marrow-stimulation alone. In the current study, we examined the modulation of cell recruitment and repair tissue characteristics at early post-surgical time points (from day 1 to 56) in a rabbit model to elucidate potential mechanisms behind this improved repair outcome. DESIGN:Thirty-three skeletally mature New Zealand White rabbits underwent bilateral arthrotomies, with each trochlea receiving a cartilage defect (3.5 mm x 4.5mm) bearing four microdrill holes (0.9 mm diameter, approximately 4 mm deep) into the subchondral bone. One defect per rabbit was treated with a chitosan-GP/blood implant, while the other defect was left as a microdrilled control. Repair tissues were stained by histochemistry, for collagen types I, II, and X by immunohistochemistry and analyzed using quantitative stereological tools. RESULTS:Histological analyses demonstrated that control defects followed a typical healing sequence observed previously in marrow-stimulation animal models while chitosan-GP/blood implants led to three significant modifications in the healing sequence at early stages: (1) increased inflammatory and marrow-derived stromal cell recruitment to the microdrill holes, (2) increased vascularization of the provisional repair tissue in the microdrill holes, and (3) increased intramembranous bone formation and subchondral bone remodeling (BR). CONCLUSIONS:These results suggest that the greater levels of provisional tissue vascularization and BR activity are main factors supporting improved cartilage repair when chitosan-GP/blood implants are applied to marrow-stimulated cartilage lesions.

背景与目标：

BACKGROUND & AIMS: :To investigate the neuronal losses of hens' spinal cords in the model of organophosphate-induced delayed neuropathy (OPIDN) and to analyze the impact of apoptosis on the pathogenesis of OPIDN. Adult hens were challenged with triorthocresyl phosphate (TOCP) at a single dose (750 mg/kg). Neuronal losses in the 3rd lumbar spinal cord (L3) were assessed by light-microscopy and electron-microscopy methods at different days post exposure, respectively. The typical OPIDN signs were seen in the TOCP-exposed hens at about 9th day. The number of large nerve cells declined gradually. And these cells were verified as neurons by immunostained with neuronal marker NeuN. The expression of FasL reached proximal at about 9th day, decreased from 14th day. Neurons in TOCP exposed groups displayed degenerative morphologies in electronic microscopy. Some neurons showed apoptotic-like ultrastructure profiles at 5th day. The nuclear membrane was complete with chromatin condensed to the margins of nuclear membrane like a crescent-shaped body. Mitochondria morphologic changes appeared early (5 d) following exposure to TOCP, and developed in a time-dependent fashion. Apoptosis might be involved in the development of OPIDN, and play a role in the pathogenesis of OPIDN.

背景与目标： : 研究有机磷酸盐诱导的迟发性神经病 (OPIDN) 模型中母鸡脊髓的神经元丢失，并分析凋亡对OPIDN发病机理的影响。以单剂量 (750 mg/kg) 对成年母鸡进行磷酸三邻甲苯酯 (TOCP) 攻击。在暴露后的不同天，分别通过光学显微镜和电子显微镜方法评估了第三腰椎脊髓 (L3) 的神经元损失。在大约第9天，在暴露于TOCP的母鸡中可以看到典型的OPIDN迹象。大神经细胞的数量逐渐减少。通过用神经元标记NeuN免疫染色将这些细胞验证为神经元。FasL的表达在第9天左右达到近端，从第14天开始下降。TOCP暴露组的神经元在电子显微镜下显示出退化形态。某些神经元在第5天显示凋亡样超微结构。核膜是完整的，染色质浓缩到核膜的边缘，就像一个新月形的身体。暴露于TOCP后早期 (5 d) 出现线粒体形态变化，并以时间依赖性方式发展。凋亡可能参与OPIDN的发生发展，并在OPIDN的发病机制中起作用。

BACKGROUND & AIMS: BACKGROUND:The purpose of this study was to determine the distribution of radioiodinated estramustine (RI-EMP) and a radioiodinated antibody against estramustine binding protein (RI-EMBP-AB) in mice. METHODS:RI-EMP and RI-EMBP-AB were injected in male mice intravenously, and the activities of tissue samples were measured 1-31 hr from the injection. Pure iodine-125 (RI) was used as a control. RESULTS:RI-EMP accumulated in the prostate, which contained 2.6% of injected activity (ID) per gram tissue at 7 hr. The liver had an activity of 21.4% ID/g at 1 hr, which decreased as RI-EMP was secreted in bile. The lung contained 2.3% ID/g at 7 hr, and it retained the activity longer than the prostate. RI-EMBP-AB accumulated in the prostate: The activity was 2.9% ID/g at 7 hr. The gallbladder contained 6.5% ID/g at 7 hr. CONCLUSIONS:Due to its cytotoxic and radiosensitizing properties, RI-EMP can possibly be used for treating prostate cancer and other tumors.

背景与目标：

BACKGROUND & AIMS: :The ATP synthase, isolated from Wolinella (formerly Vibrio) succinogenes could be fully incorporated into liposomes without significant cleavage of the enzyme or loss of activity. These proteoliposomes, but not the isolated enzyme, catalyzed phosphate-ATP exchange and the phosphorylation of ADP which was driven by an artificially imposed delta mu H across the liposomal membrane. Phosphorylation driven by light was catalyzed by proteoliposomes containing also bacteriorhodopsin. The three activities were similarly sensitive to protonophores or dicyclohexylcarbodiimide. This sensitivity was similar to that of the electron-transport-driven phosphorylation catalyzed by bacterial membrane vesicles. With a delta mu H value 280 mV to drive phosphorylation the turnover number of the enzyme was in the same order of magnitude as that measured in the electron-transport-driven phosphorylation catalyzed by the bacterial membrane. When the delta mu H was below 150 mV, the phosphorylation activity of the incorporated enzyme was two orders of magnitude slower, and was about as fast as light-driven phosphorylation or as the exchange reaction.

背景与目标： : 从Wolinella (以前称为弧菌) 琥珀生成素中分离出的ATP合酶可以完全掺入脂质体中，而不会显着裂解酶或丧失活性。这些蛋白脂质体 (而不是分离的酶) 催化了磷酸盐-ATP交换和ADP的磷酸化，而ADP的磷酸化是由在脂质体膜上人工施加的 δ mu H驱动的。光驱动的磷酸化是由也含有细菌视紫红质的蛋白脂质体催化的。这三种活性对原蛋白或二环己基碳二亚胺敏感。这种敏感性与细菌膜囊泡催化的电子传输驱动的磷酸化相似。在280 mV驱动磷酸化的 δ μ H值的情况下，酶的周转数与在由细菌膜催化的电子传输驱动的磷酸化中测量的周转数处于相同的数量级。当 δ μ H低于150 mV时，掺入的酶的磷酸化活性慢两个数量级，并且大约与光驱动磷酸化或交换反应一样快。

BACKGROUND & AIMS: :Inorder to brought S-naproxen into small intestine, an optically pure (S)-naproxen starch ester was produced by lipase through enantio-selective trans-esterification of racemic naproxen methyl ester with pretreatment starch in solvent system. With carefully selection of the reaction medium (isooctane), lipase (Carica Papaya Lipase, CPL) and the reaction mode (intermittent opening), a high conversion rate (48.6%) and enantiomeric excess of product (99.6%) was obtained. The slow release macromolecular (S)-Naproxen had been synthesized to improve the efficacy of racemic naproxen and overcome its side effects. The enanitomeric ratio of CPL (E=52.5) was higher than CRL (E=22) and greatly influenced by the byproduct methyl alcohol. The intermittent opening reaction mode was the effective way to remove the inhibition of methyl alcohol and to improve the enantio-selectivity of CPL. S-naproxen starch was confirmed by HPLC and 1H NMR. This method may also apply to preparation the other optically pure 2-phenylpropionic acid derivatives. S-naproxen starch was a new optically pure derivatives possessing emulsifying and slow release properties would be widely applied to the food, pharmaceutical and biomedical industries.

背景与目标： : 为了将S-萘普生带入小肠，通过脂肪酶通过外消旋萘普生甲酯与预处理淀粉在溶剂体系中的对映选择性酯化反应，制得光学纯的 (S)-萘普生淀粉酯。通过仔细选择反应介质 (异辛烷) 、脂肪酶 (番木瓜脂肪酶，CPL) 和反应模式 (间歇开放)，获得高转化率 (48.6%) 和对映体过量的产物 (99.6%)。合成了缓释大分子萘普生，以提高外消旋萘普生的功效并克服其副作用。CPL的烯醇比 (E = 52.5) 高于CRL (E = 22)，并且受副产物甲醇的影响很大。间歇开放反应模式是去除甲醇抑制和提高CPL对映体选择性的有效途径。通过HPLC和1H NMR证实了S-萘普生淀粉。该方法也可用于制备其他光学纯的2-苯基丙酸衍生物。S-萘普生淀粉是一种新型的光学纯衍生物，具有乳化和缓释特性，被广泛应用于食品、制药和生物医药行业。