BACKGROUND & AIMS: Interleukin-1beta (IL-1beta) is closely involved in liver disorders. IL-1beta produces nitric oxide (NO) in vascular smooth muscle cells and relaxes vascular smooth muscle via cyclic guanosine 3',5'-monophosphate (cGMP). In this study, we evaluated the relaxing effect of IL-1beta on cultured Ito cells. Ito cells were isolated from the livers of male Wistar rats and cultured for 24 hours. Immunolocalization of inducible nitric oxide synthase (iNOS) and cGMP and intensity of fluorescence of cGMP were examined using a confocal laser microscope. Ito cells were treated with 0, 200, and 1,000 pmol/L IL-1beta, and the intracellular cGMP concentration was measured after 12 hours. Moreover, Ito cells treated with 200 and 1,000 pmol/L IL-1beta and not treated with IL-1beta were observed over 12 hours, and the area of the same Ito cell was compared before and after the addition of IL-1beta. Next, effects of N(G)-monomethyl-L-arginine (L-NMMA) and S-nitroso-N-acetyl-DL-penicillamine (SNAP) on Ito cell relaxation by IL-1beta treatment were examined. In Ito cells, immunofluorescence of iNOS was observed, and fluorescent intensity of cGMP increased after addition of IL-1beta. Intracellular cGMP concentration increased dose-dependently after addition of IL-1beta. Cell area significantly increased in the IL-1beta-treated group compared with the untreated group. Relaxation of Ito cells by IL-1beta treatment was inhibited by L-NMMA in a dose-dependent manner, but was enhanced by SNAP. These results indicate that IL-1beta produces NO in cultured Ito cells and relaxes the cells via cGMP.

背景与目标： 白介素-1β（IL-1beta）与肝脏疾病密切相关。 IL-1beta通过环状鸟苷3'，5'-单磷酸（cGMP）在血管平滑肌细胞中产生一氧化氮（NO），并使血管平滑肌松弛。在这项研究中，我们评估了IL-1β对培养的Ito细胞的松弛作用。从雄性Wistar大鼠的肝脏中分离出Ito细胞，并培养24小时。使用共聚焦激光显微镜检查诱导型一氧化氮合酶（iNOS）和cGMP的免疫定位和cGMP的荧光强度。用0、200和1,000 pmol / L IL-1beta处理Ito细胞，并在12小时后测量细胞内cGMP浓度。此外，在12小时内观察到用200和1,000 pmol / L IL-1beta处理且未用IL-1beta处理的Ito细胞，并且在添加IL-1beta之前和之后比较了同一个Ito细胞的面积。接下来，研究了N（G）-单甲基-L-精氨酸（L-NMMA）和S-亚硝基-N-乙酰基-DL-青霉胺（SNAP）对通过IL-1beta处理的Ito细胞松弛的影响。在Ito细胞中，观察到iNOS的免疫荧光，添加IL-1β后cGMP的荧光强度增加。加入IL-1beta后，细胞内cGMP浓度呈剂量依赖性增加。与未治疗组相比，IL-1β治疗组的细胞面积显着增加。 L-1NMMA以剂量依赖的方式抑制了通过IL-1beta处理的Ito细胞的松弛，但被SNAP增强了。这些结果表明IL-1β在培养的Ito细胞中产生NO，并通过cGMP使细胞松弛。

BACKGROUND & AIMS: :The signaling pathways mediating lysophosphatidic acid (LPA)-stimulated PKD(2) activation and the potential contribution of PKD(2) in regulating LPA-induced interleukin 8 (IL-8) secretion in nontransformed, human colonic epithelial NCM460 cells were examined. Treatment of serum-deprived NCM460 cells with LPA led to a rapid and striking activation of PKD(2), as measured by in vitro kinase assay and phosphorylation at the activation loop (Ser706/710) and autophosphorylation site (Ser876). PKD(2) activation induced by LPA was abrogated by preincubation with selective PKC inhibitors GF-I and Ro-31-8220 in a dose-dependent manner. These inhibitors did not have any direct inhibitory effect on PKD(2) activity. LPA induced a striking increase in IL-8 production and stimulated NF-kappaB activation, as measured by NF-kappaB-DNA binding, NF-kappaB-driven luciferase reporter activity, and IkappaBalpha phosphorylation. PKD(2) gene silencing utilizing small interfering RNAs targeting distinct PKD(2) sequences dramatically reduced LPA-stimulated NF-kappaB promoter activity and IL-8 production. PKD(2) activation is a novel early event in the biological action of LPA and mediates LPA-stimulated IL-8 secretion in NCM460 cells through a NF-kappaB-dependent pathway. Our results demonstrate, for the first time, the involvement of a member of the PKD family in the production of IL-8, a potent proinflammatory chemokine, by epithelial cells.

背景与目标： ：审查了介导溶血磷脂酸（LPA）刺激的PKD（2）激活和PKD（2）在调节LPA诱导的非转化型人结肠上皮NCM460细胞中LPA诱导的白介素8（IL-8）分泌中的潜在作用。用体外LPA处理血清缺乏的NCM460细胞会导致PKD（2）迅速而惊人的活化，这是通过体外激酶测定和活化环（Ser706 / 710）和自磷酸化位点（Ser876）的磷酸化来测量的。通过与选择性PKC抑制剂GF-1和Ro-31-8220呈剂量依赖性方式进行预孵育，可以消除LPA诱导的PKD（2）激活。这些抑制剂对PKD（2）活性没有任何直接的抑制作用。如通过NF-κB-DNA结合，NF-κB驱动的萤光素酶报道分子活性和IkappaBalpha磷酸化所测量，LPA诱导IL-8产量显着增加并刺激NF-κB活化。 PKD（2）基因沉默利用针对不同的PKD（2）序列的小干扰RNA，大大降低了LPA刺激的NF-κB启动子活性和IL-8的产生。 PKD（2）激活是LPA的生物学作用中的一个新的早期事件，并通过NF-κB依赖性途径介导NCM460细胞中LPA刺激的IL-8分泌。我们的结果首次证明，上皮细胞参与了PKD家族成员参与IL-8（一种有效的促炎趋化因子）的生产。

BACKGROUND & AIMS: :The gram-negative bacterium Bartonella henselae is capable of causing angiogenic lesions as a result of infection. Previously, it has been shown that B. henselae infection can result in production of the chemokine interleukin-8 (IL-8). In this study, we demonstrated that monocytes, endothelial cells, and hepatocytes produce IL-8 in response to B. henselae infection. We also investigated the role of IL-8 in B. henselae-induced endothelial cell proliferation and capillary tube formation. Both in vitro angiogenesis assays were IL-8 dependent. B. henselae-mediated inhibition of apoptosis, as indicated by gene expression of Bax and Bcl-2, was also shown to be IL-8 dependent in endothelial cells. Furthermore, infection of endothelial cells with B. henselae stimulated upregulation of the IL-8 chemokine receptor CXCR2. Infection of human endothelial cells by B. henselae resulting in IL-8 production likely plays a central role in the ability of this organism to cause angiogenesis during infection.

背景与目标： 革兰氏阴性细菌汉氏巴尔通体（Bartonella henselae）能够由于感染而引起血管生成性病变。以前，已经证明，亨氏芽孢杆菌感染可以导致趋化因子白介素8（IL-8）的产生。在这项研究中，我们证明了单核细胞，内皮细胞和肝细胞对B. henselae感染产生IL-8。我们还研究了IL-8在B. henselae诱导的内皮细胞增殖和毛细管形成中的作用。两种体外血管生成测定都是IL-8依赖性的。如Bax和Bcl-2的基因表达所表明的，B。henselae介导的凋亡抑制在内皮细胞中也显示为IL-8依赖性。此外，用汉逊芽孢杆菌感染内皮细胞刺激了IL-8趋化因子受体CXCR2的上调。亨氏芽孢杆菌对人内皮细胞的感染导致IL-8的产生可能在该生物体在感染过程中引起血管生成的能力中起着核心作用。

BACKGROUND & AIMS: :Epithelial cells play an important role in orchestrating mucosal immune responses. In allergic-type inflammation, epithelial cells control the recruitment of eosinophils into the mucosa. Th2-type cytokine-driven release of eosinophil-active chemokines from epithelial cells directs eosinophil migration into the mucosal epithelium. CCR3, the main eosinophil chemokine receptor, regulates this process; however, the respective contribution of individual CCR3 ligands in eosinophil transepithelial migration is less well understood. Using an in vitro transepithelial chemotaxis system, we found that eotaxin-3 produced by IL-4-stimulated airway epithelial cells and CCR3 on eosinophils exclusively mediate eosinophil transepithelial migration. Eotaxin-3 protein levels were also increased in the nasal mucosal epithelium recovered from allergic patients as compared to non-allergic patients. Surprisingly, eotaxin-3 in IL-4-stimulated airway epithelial cells was predominantly cell surface bound, and the cell surface form was critical for eosinophil transepithelial migration. Eotaxin-3 cell surface association was partially glycosaminoglycan (GAG) dependent, but was completely protein dependent, suggesting that eotaxin-3 associates with both GAG and cell surface proteins. We thus provide evidence that cell surface-associated eotaxin-3 is the critical IL-4-dependent chemotactic signal mediating eosinophil transepithelial migration in the setting of allergic inflammation.

背景与目标： ：上皮细胞在协调粘膜免疫反应中起重要作用。在过敏型炎症中，上皮细胞控制嗜酸性粒细胞向粘膜的募集。 Th2型细胞因子驱动的嗜酸性粒细胞活性趋化因子从上皮细胞的释放引导嗜酸性粒细胞迁移到粘膜上皮中。 CCR3是主要的嗜酸性粒细胞趋化因子受体，调节这一过程。然而，单个CCR3配体在嗜酸性粒细胞上皮细胞迁移中的各自作用尚不清楚。使用体外经上皮趋化系统，我们发现由IL-4刺激的气道上皮细胞和嗜酸性粒细胞上的CCR3产生的eotaxin-3专门介导嗜酸性粒细胞跨上皮迁移。与非变态反应患者相比，从变态反应患者回收的鼻粘膜上皮中的Eotaxin-3蛋白水平也升高。出人意料的是，IL-4刺激的气道上皮细胞中的eotaxin-3主要是细胞表面结合的，并且细胞表面形式对于嗜酸性粒细胞跨上皮迁移至关重要。嗜酸性粒细胞趋化因子3（Eotaxin-3）细胞表面缔合部分依赖糖胺聚糖（GAG），但完全依赖蛋白质，这表明嗜酸性粒细胞趋化因子3（Eotaxin-3）与GAG和细胞表面蛋白缔合。因此，我们提供证据，在过敏性炎症的情况下，细胞表面相关的eotaxin-3是介导嗜酸性粒细胞跨上皮迁移的关键IL-4依赖性趋化信号。

BACKGROUND & AIMS: A primer extension (toeprinting) assay was used to monitor selection by ribosomes of the first versus the second AUG codon as a function of introducing mutations on the 3' side (positions +4, +5 and +6) of the first AUG codon. Six different flanking codons starting with G (GCG, GCU, GCC, GCA, GAU and GGA) strongly augmented selection of AUG#1 when compared with matched mRNAs that had A or C instead of G in position +4. Augmentation by G in position +4 failed only when it was combined with U in position +5, as in the sequence augGUA. In contrast with the usual enhancing effect of introducing G in position +4, most mutations in position +5 had no discernible effect, as shown with the series augANA (where N = C, A, G or U) and the series augCNA. AUG codon recognition was also unaffected by mutations in position +6, as shown by testing four mRNAs that had augCCN as the start site. Thus the primary sequence context that augments the recognition of AUG start codons does not appear generally to extend beyond G in position +4. When the toeprinting assay was used with mRNAs that initiate translation at CUG instead of AUG, cugGAU was not recognized better than cugGGU, contradicting the hypothesis that initiation at non-AUG codons might be favored by A instead of G in position +5.

背景与目标： 使用引物延伸（印迹）测定法来监测第一和第二AUG密码子的核糖体的选择，其作为在第一AUG密码子的3'侧（位置4、5和6）上引入突变的函数。与匹配的在位置4有A或C而不是G的mRNA相比，以G开头的六个不同的侧翼密码子（GCG，GCU，GCC，GCA，GAU和GGA）大大增强了对AUG＃1的选择。仅当它与位置5的U组合时失败，如序列augGUA所示。与在位置4引入G的通常增强作用相反，在位置5的大多数突变没有明显的作用，如augANA系列（其中N = C，A，G或U）和augCNA系列所示。 AUG密码子识别也不受位置6突变的影响，如测试以augCCN为起始位点的四个mRNA所显示的。因此，增强对AUG起始密码子识别的一级序列通常不会延伸到位置4的G范围之外。当将脚印法与在CUG而非AUG处起始翻译的mRNA一起使用时，cugGAU的识别度不如cugGGU，与以下假设相矛盾：在非AUG密码子处的起始可能会被位置5的A而不是G所偏爱。

BACKGROUND & AIMS: Interleukin-1 (IL-1) has been implicated in chronic and acute cerebral neuropathologies. IL-1 receptor antagonist (IL-1ra), a naturally occurring protein that binds to IL-1 receptors without inducing signal transduction, blocks several actions of IL-1. IL-1ra acts at the local level and it also circulates in the bloodstream. We now report evidence for a biological function of IL-1ra in the brain as an endogenous neuroprotective molecule. Cerebral expression of IL-1ra mRNA is induced rapidly by focal cerebral ischemia in rats, and inhibition of the action of IL-1ra, by passive immuno-neutralization, markedly enhances ischemic damage. To our knowledge this is the first report of an action of endogenous IL-1ra in the brain. Control of IL-1ra expression or action may therefore provide a useful therapeutic strategy to limit acute neurodegeneration.

背景与目标： 白介素-1（IL-1）已牵涉到慢性和急性脑神经病理学。 IL-1受体拮抗剂（IL-1ra）是一种与IL-1受体结合而不诱导信号转导的天然蛋白质，可阻断IL-1的多种作用。 IL-1ra在局部起作用，并且也在血液中循环。我们现在报告IL-1ra在脑中作为内源性神经保护分子的生物学功能的证据。大鼠局灶性脑缺血可快速诱导IL-1ra mRNA的脑表达，并且被动免疫中和抑制IL-1ra的作用可显着增强缺血性损伤。据我们所知，这是大脑中内源性IL-1ra作用的首次报道。因此，控制IL-1ra的表达或作用可能为限制急性神经变性提供了有用的治疗策略。

BACKGROUND & AIMS: :A structure-activity relationship (SAR) study was performed principally at the N1 position of N1-arylsulfonyl-N2-[1-(1-naphthyl)ethyl]-trans-1,2-diaminocyclohexanes, a new family of calcilytics acting at the calcium sensing receptor (CaSR). The most active compound in this series was the 4-(trifluoromethoxy)benzenesulfonyl derivative 7e, which displayed an IC50 of 5.4 +/- 0.5 microM with respect to the inhibition of calcium-induced tritiated inositol phosphate ([3H]IP) accumulation in Chinese hamster ovarian (CHO) cells expressing the CaSR. Replacement of the sulfonamide linkage of this compound by a carboxamide led to a 6-fold increase in activity (7m, IC50 = 0.9 +/- 0.2 microM). Among the carboxamides synthesized, one of the most active compounds was the 4-chlorophenylcarboxamide (1S,2S,1'R)-7n (Calhex 231, IC50 = 0.33 +/- 0.02 microM). The absolute configuration of (1S,2S,1'R)-7n was deduced from an X-ray crystallographic study of one of the diastereomers of compound 7d. The stereochemical preference for the (1S,2S,1'R)-isomers can be rationalized on the basis of a three-dimensional model of the calcilytic binding pocket of the CaSR. Removal of the C-1' methyl group or replacement of the 1-naphthyl group by a 2-naphthyl or biphenyl moiety led to appreciable loss of calcilytic activity. Compounds 7e, 7m, and Calhex 231 did not stimulate [3H]IP accumulation in CHO cells expressing or not expressing the CaSR.

背景与目标： ：结构活性关系（SAR）研究主要在N1-芳基磺酰基-N2- [1-（1-（萘基）乙基]-反式1,2-二氨基环己烷的N1位置进行的钙敏感受体（CaSR）。该系列中活性最高的化合物是4-（三氟甲氧基）苯磺酰基衍生物7e，在抑制中国仓鼠中钙诱导的tri化肌醇磷酸酯（[3H] IP）积累方面，其IC50为5.4 /-0.5 microM。表达CaSR的卵巢（CHO）细胞。该化合物的磺酰胺键被羧酰胺取代导致活性增加了6倍（7m，IC50 = 0.9 /-0.2 microM）。在合成的羧酰胺中，活性最高的化合物之一是4-氯苯基羧酰胺（1S，2S，1'R）-7n（Calhex 231，IC50 = 0.33 /-0.02 microM）。 （1S，2S，1'R）-7n的绝对构型是通过对化合物7d的一种非对映异构体进行X射线晶体学研究得出的。 （1S，2S，1'R）-异构体的立体化学偏好可以基于CaSR钙解结合口袋的三维模型来合理化。除去C-1'甲基或用2-萘基或联苯部分取代1-萘基导致明显的钙解活性损失。化合物7e，7m和Calhex 231不会刺激表达或不表达CaSR的CHO细胞中的[3H] IP积累。

BACKGROUND & AIMS: :The TIM (T cell/transmembrane, immunoglobulin and mucin) proteins are crucial regulators of Th1/Th2 immune responses and have been implicated in several diseases including HIV-1/AIDS. The TIM1 exon 4 that codes for mucin domain is highly diverse, with sequence variants associated with varying phenotypes. In this study, TIM1 exon 4 was sequenced among 227 HIV-1 seroprevalent and 288 healthy non infected individuals from North Indian population and haplotypes established. A novel but rare haplotype D1(∗) was identified among the healthy and differed from D1 by a synonymous substitution G>T at Thr208Thr. The TIM1 haplotype diversity showed no association with susceptibility to HIV-1 infection. The seroprevalent individuals carrying D3A had relatively higher median CD4+T cell counts (368/μl) than those without (313/μl; p=0.02). A comparison of CD4+T counts between D3-A individuals on ART or ART naïve did not show any significant difference plausibly due to confounding nature of ART and other factors.

背景与目标： TIM（T细胞/跨膜，免疫球蛋白和粘蛋白）蛋白是Th1 / Th2免疫反应的关键调节剂，并与包括HIV-1 / AIDS在内的多种疾病有关。编码粘蛋白结构域的TIM1外显子4高度多样化，具有与不同表型相关的序列变体。在这项研究中，TIM1外显子4在来自北印度人口的227个HIV-1血清流行和288个健康的未感染个体中进行了测序，并建立了单倍型。在健康人群中鉴定出一种新颖但罕见的单倍型D1（∗），与D1的区别在于在Thr208Thr处的同义替代G> T。 TIM1单倍型多样性表明与HIV-1感染的易感性无关。携带D3A的血清流行个体的CD4 T细胞计数中位数相对较高（不含313 /μl； p = 0.02）。由于ART和其他因素的混杂，D3-A个体或未接受过ART的D3-A个体之间CD4 T计数的比较似乎没有显示任何显着差异。

BACKGROUND & AIMS: :Abstract Type III interferon (IFN-λ) is a novel member of the interferon family, which preferentially promotes antiviral responses from epithelial cells and cooperates with type I IFNs in the clearance of viral infections. However, the effect of mIFN-λ2 to the LA795 lung adenocarcinoma cell is largely unknown. In this study, we transfected Ad-mIFN-λ2 vector into LA795 tumor-bearing mice to explore the effect of mIFN-λ2 on the proliferation of LA795 lung adenocarcinoma cell and on the immune response of the mice. Transfected by Ad-mIFN-λ2 vector, a significant decrease in the tumor growth, the subcutaneous tumor necrosis, cystic degeneration, and tumor apoptosis were more evident; at the same time, mIFN-λ2 protein and gene were significantly more expressed. And, flow cytometry analysis suggested that CD3(+)CD4(+), CD3(+)CD8(+), and NK (CD3(-)CD49(+)) cells were all significantly increased after transfected by Ad-mIFN-λ2. The study demonstrated that recombinant Ad-mIFN-λ2 transfection effectively inhibited the growth of LA795 lung adenocarcinoma cell, which may work through inducing apoptosis of tumor cell and regulating cell immune response.

背景与目标： 摘要：III型干扰素（IFN-λ）是干扰素家族的一个新成员，它优先促进上皮细胞的抗病毒反应，并与I型干扰素协同作用以清除病毒感染。然而，很大程度上未知mIFN-λ2对LA795肺腺癌细胞的作用。在这项研究中，我们将Ad-mIFN-λ2载体转染到LA795荷瘤小鼠中，以研究mIFN-λ2对LA795肺腺癌细胞增殖和小鼠免疫反应的影响。 Ad-mIFN-λ2载体转染后，肿瘤生长明显减少，皮下肿瘤坏死，囊性变性和肿瘤细胞凋亡更为明显。同时，mIFN-λ2蛋白和基因的表达明显增加。并且，流式细胞仪分析表明，用Ad-mIFN-λ2转染后，CD3（）CD4（），CD3（）CD8（）和NK（CD3（-）CD49（））细胞均显着增加。研究表明重组Ad-mIFN-λ2转染有效抑制了LA795肺腺癌细胞的生长，其作用可能是通过诱导肿瘤细胞凋亡和调节细胞免疫应答来实现的。

BACKGROUND & AIMS: :Reported here is the synthesis and biological evaluation of the asialoglycoprotein receptor (ASGP-R) targeted fourth generation poliamidoamine dendrimer (G(4)-PAMAM) loaded with sorafenib. The ASGP-R targeted dendrimer was obtained by conjugation of Lactobionic acid (La) to the G(4)-PAMAM dendrimer, followed by acetylation (Ac) of the free amino groups in order to reduce the non-specific interactions with the cell membrane. Moreover, by additionally grafting fluorescein (FITC), it was easy to characterize the internalization pathway and the intracellular fate of the targeted dendrimer Ac-La-G(4)-PAMAM-FITC. In vitro experiments performed on HepG-2 and HLE cell lines, allowed to study the ability of the dendrimers to affect the cell vitality. Confocal microscopy and cytofluorimetric analysis confirmed higher binding and uptake ability of the Ac-La-G(4)-PAMAM-FITC dendrimer in well differentiated and ASGP-R expressing human liver cancer cell line HepG-2 compared non-expressing HLE cells. Ac-La-G(4)-PAMAM-FITC dendrimer loaded with sorafenib was stable and showed sustained sorafenib release. As evidenced by the cytotoxicity studies, sorafenib included in the dendrimer maintained its effectiveness, and was able to produce a longer lasting effect over the time compared to molar equivalent doses of free sorafenib. This new targeted dendrimer appears to be a suitable carrier for the delivery of sorafenib to liver cancer cells expressing ASGP-R.

背景与目标： ：此处报道的是载有索拉非尼的去唾液酸糖蛋白受体（ASGP-R）靶向的第四代多巴胺胺树状大分子（G（4）-PAMAM）的合成和生物学评估。 ASGP-R靶向树状聚合物是通过将乳酸基乳酸（La）与G（4）-PAMAM树状聚合物缀合，然后对游离氨基进行乙酰化（Ac），以减少与细胞膜的非特异性相互作用而获得的。此外，通过额外嫁接荧光素（FITC），很容易表征目标树状聚合物Ac-La-G（4）-PAMAM-FITC的内在途径和细胞内命运。在HepG-2和HLE细胞系上进行的体外实验可以研究树状聚合物影响细胞活力的能力。共聚焦显微镜和细胞荧光分析证实，在未分化的HLE细胞中，Ac-La-G（4）-PAMAM-FITC树状大分子在高分化和表达ASGP-R的人肝癌细胞系HepG-2中具有更高的结合和摄取能力。载有索拉非尼的Ac-La-G（4）-PAMAM-FITC树状聚合物稳定并显示索拉非尼持续释放。正如细胞毒性研究所证明的那样，与摩尔当量剂量的游离索拉非尼相比，树状大分子中包含的索拉非尼保持了其有效性，并能够在一段时间内产生更长的持久作用。这种新的靶向树状聚合物看来是将索拉非尼递送至表达ASGP-R的肝癌细胞的合适载体。

BACKGROUND & AIMS: :Type 2 diabetes is a strong risk factor for stroke. Linagliptin is a dipeptidyl peptidase-4 (DPP-4) inhibitor in clinical use against type 2 diabetes. The aim of this study was to determine the potential antistroke efficacy of linagliptin in type 2 diabetic mice. To understand whether efficacy was mediated by glycemia regulation, a comparison with the sulfonylurea glimepiride was done. To determine whether linagliptin-mediated efficacy was dependent on a diabetic background, experiments in nondiabetic mice were performed. Type 2 diabetes was induced by feeding the mice a high-fat diet for 32 weeks. Mice were treated with linagliptin/glimepiride for 7 weeks. Stroke was induced at 4 weeks into the treatment by transient middle cerebral artery occlusion. Blood DPP-4 activity, glucagon-like peptide-1 (GLP-1) levels, glucose, body weight, and food intake were assessed throughout the experiments. Ischemic brain damage was measured by determining stroke volume and by stereologic quantifications of surviving neurons in the striatum/cortex. We show pronounced antistroke efficacy of linagliptin in type 2 diabetic and normal mice, whereas glimepiride proved efficacious against stroke in normal mice only. These results indicate a linagliptin-mediated neuroprotection that is glucose-independent and likely involves GLP-1. The findings may provide an impetus for the development of DPP-4 inhibitors for the prevention and treatment of stroke in diabetic patients.

背景与目标： ：2型糖尿病是中风的重要危险因素。 Linagliptin是临床上用于治疗2型糖尿病的二肽基肽酶4（DPP-4）抑制剂。这项研究的目的是确定利格列汀在2型糖尿病小鼠中的潜在抗中风功效。为了了解功效是否由血糖调节介导，与磺酰脲格列美脲进行了比较。为了确定利格列汀介导的功效是否取决于糖尿病背景，在非糖尿病小鼠中进行了实验。通过给小鼠喂高脂饮食32周来诱发2型糖尿病。用利格列汀/格列美脲治疗小鼠7周。在治疗的第4周，通过短暂性中脑动脉闭塞诱发中风。在整个实验过程中，评估了血液DPP-4活性，胰高血糖素样肽1（GLP-1）水平，葡萄糖，体重和食物摄入量。通过测定中风量和纹状体/皮层中存活的神经元的立体定量来测量缺血性脑损伤。我们在2型糖尿病和正常小鼠中显示了利格列汀的显着抗中风功效，而格列美脲仅在正常小鼠中被证明对中风有效。这些结果表明，由利格列汀介导的神经保护作用与葡萄糖无关，并且可能涉及GLP-1。这些发现可能为DPP-4抑制剂的开发提供动力，以预防和治疗糖尿病患者的中风。

BACKGROUND & AIMS: :Agents to control melanogenesis are in demand for the development of cosmetics to improve pigmentation disorders of skin and hair. In this study, we examined and evaluated the effects of flavonoids on melanogenesis in the melanogenic cells model, murine B16F10 melanoma cells. In the course of this study, we found that incubation of the cells in a medium containing 10 μM of the 4'-O-methylated flavonoids, diosmetin (4'-O-methylluteolin), acacetin (4'-O-methylapigenin) or kaempferide (4'-O-methylkaempferol), increased the melanin contents of the cells 3- to 7-fold higher than the control cells. The concentration-dependence test revealed that 20 μM acacetin showed the highest effect, up to 33-fold higher than the vehicle. On the other hand, the corresponding 4'-OH-type flavonoids, luteolin, apigenin and kaempferol, had a significantly smaller effect. Furthermore, by evaluating the melanogenic proteins, we found that the cells treated with 4'-O-methylated flavonoids showed higher tyrosinase activity, as well as upregulation of tyrosinase expression, preceded by activation of cAMP response element binding protein (CREB) and extracellular signal-regulated kinases types 1 and 2 (ERK1/2). These results indicate that the 4'-O-methyl group of flavonoids plays an important role in the induction of melanogenesis by activating its major signal transduction pathway through the upregulation of phospho-CREB in murine B16F10 melanoma cells.

背景与目标： ：需要控制黑素生成的试剂来开发化妆品，以改善皮肤和头发的色素沉着障碍。在这项研究中，我们检查和评估了类黄酮对黑色素生成细胞模型鼠B16F10黑色素瘤细胞中黑色素生成的影响。在这项研究过程中，我们发现细胞在含有10μM4'-O-甲基化类黄酮，薯os皂素（4'-O-甲基木犀草素），阿沙西汀（4'-O-甲基芹菜素）或Kaempferide（4'-O-methylkaempferol）使细胞的黑色素含量比对照细胞高3至7倍。浓度依赖性试验表明，20μM醋氨蝶呤显示出最高的作用，比赋形剂高33倍。另一方面，相应的4'-OH型黄酮，木犀草素，芹菜素和山ka酚的作用明显较小。此外，通过评估黑色素生成蛋白，我们发现用4'-O-甲基化类黄酮处理的细胞显示出更高的酪氨酸酶活性，以及​​酪氨酸酶表达的上调，然后激活cAMP反应元件结合蛋白（CREB）和细胞外信号调节的1型和2型激酶（ERK1 / 2）。这些结果表明，类黄酮的4'-O-甲基通过激活鼠B16F10黑色素瘤细胞中的磷酸-CREB激活其主要信号转导途径，从而在诱导黑色素生成中起重要作用。

BACKGROUND & AIMS: :Utilizing X-ray crystal structure analysis, (3S,5R)-5-[4-(2-chlorophenyl)-2,2-dimethyl-5-oxopiperazin-1-yl]piperidine-3-carboxamides were designed and identified as renin inhibitors. The most potent compound 15 demonstrated favorable pharmacokinetic and pharmacodynamic profiles in rat.

背景与目标： ：利用X射线晶体结构分析，设计了（3S，5R）-5- [4-（2-氯苯基）-2,2-二甲基-5-氧哌嗪-1-基]哌啶-3-甲酰胺，并将其鉴定为肾素抑制剂。最有效的化合物15在大鼠中显示出良好的药代动力学和药效学特征。

BACKGROUND & AIMS: :Antitumor agents that bind to tubulin and disrupt microtubule dynamics have attracted considerable attention in the last few years. To extend our knowledge of the thiazole ring as a suitable mimic for the cis-olefin present in combretastatin A-4, we fixed the 3,4,5-trimethoxyphenyl at the C4-position of the thiazole core. We found that the substituents at the C2- and C5-positions had a profound effect on antiproliferative activity. Comparing compounds with the same substituents at the C5-position of the thiazole ring, the moiety at the C2-position influenced antiproliferative activities, with the order of potency being NHCH(3) > Me > N(CH(3))(2). The N-methylamino substituent significantly improved antiproliferative activity on MCF-7 cells with respect to C2-amino counterparts. Increasing steric bulk at the C2-position from N-methylamino to N,N-dimethylamino caused a 1-2 log decrease in activity. The 2-N-methylamino thiazole derivatives 3b, 3d and 3e were the most active compounds as antiproliferative agents, with IC(50) values from low micromolar to single digit nanomolar, and, in addition, they are also active on multidrug-resistant cell lines over-expressing P-glycoprotein. Antiproliferative activity was probably caused by the compounds binding to the colchicines site of tubulin polymerization and disrupting microtubule dynamics. Moreover, the most active compound 3e induced apoptosis through the activation of caspase-2, -3 and -8, but 3e did not cause mitochondrial depolarization.

背景与目标： 近年来，与微管蛋白结合并破坏微管动力学的抗肿瘤剂引起了相当大的关注。为了扩展我们对噻唑环作为康美他汀A-4中存在的顺式烯烃的合适模拟物的认识，我们将3,4,5-三甲氧基苯基固定在噻唑核心的C4位。我们发现，在C2和C5位置的取代基对抗增殖活性具有深远的影响。比较噻唑环C5位上具有相同取代基的化合物，C2位上的部分影响抗增殖活性，效力顺序为NHCH（3）> Me> N（CH（3））（2） 。相对于C 2-氨基对应物，N-甲基氨基取代基显着提高了对MCF-7细胞的抗增殖活性。从N-甲基氨基到N，N-二甲基氨基的C2位增加的空间体积导致活性降低1-2 log。 2-N-甲基氨基噻唑衍生物3b，3d和3e是最有效的化合物作为抗增殖剂，其IC（50）值从低微摩尔到一位数纳摩尔，此外，它们还对耐多药细胞具有活性系过度表达P-糖蛋白。抗增殖活性可能是由于化合物与微管蛋白聚合的秋水仙碱位点结合并破坏了微管动力学而引起的。此外，活性最高的化合物3e通过激活caspase-2，-3和-8诱导细胞凋亡，但3e不会引起线粒体去极化。