BACKGROUND & AIMS: :Heat shock protein 90 (HSP90) is required for structural folding and maintenance of conformational integrity of various proteins, including several associated with cellular signaling. Recent studies utilizing 17-allylamino-17-demethoxygeldanamycin (17-AAG), an inhibitor of HSP90, demonstrated an antitumor effect in solid tumors. To test whether HSP90 could be targeted in multiple myeloma (MM) patients, we first investigated expression of HSP90 by immunofluorescence and flow cytometric analysis in a myeloma cell line (U266) and primary myeloma cells. Following demonstration of HSP90 expression in myeloma cells, archival samples of 32 MM patients were analysed by immunoperoxidase staining. Myeloma cells in all patients showed strong cytoplasmic expression of HSP90 in all samples and 55% also demonstrated concurrent nuclear immunopositivity. Treatment of U266 and primary MM cells with 17AAG resulted in significantly increased apoptosis compared to untreated control cells. Analysis of anti-apoptotic BCL2 family proteins and akt in MM cells incubated with 17-AAG revealed down-regulation of BCL-2, BCL-XL, MCL-1 and akt. Furthermore, although a low concentration of bortezomib resulted in no cell death, a combination of 17AAG and bortezomib treatment revealed a synergistic apoptotic effect on the U266 cell line. These data suggest that targeted inhibition of HSP90 may prove to be a valid and innovative strategy for the development of future therapeutic options for MM patients.

背景与目标： : 热休克蛋白90 (HSP90) 是各种蛋白质的结构折叠和构象完整性维持所必需的，包括与细胞信号传导相关的几种。利用HSP90抑制剂17-allylamino-17-demethoxygeldanamycin (17-AAG) 的最新研究表明，在实体瘤中具有抗肿瘤作用。为了测试HSP90是否可以靶向多发性骨髓瘤 (MM) 患者，我们首先通过免疫荧光和流式细胞仪分析研究了HSP90在骨髓瘤细胞系 (U266) 和原发性骨髓瘤细胞中的表达。在证明骨髓瘤细胞中HSP90表达后，通过免疫过氧化物酶染色分析了32 MM患者的档案样本。所有患者的骨髓瘤细胞在所有样品中均显示出HSP90的强细胞质表达，并且55% 还显示出同时的核免疫阳性。与未处理的对照细胞相比，用17AAG处理U266和原代MM细胞可显着增加细胞凋亡。对与17-aag孵育的MM细胞中抗凋亡BCL2家族蛋白和akt的分析表明，BCL-2，BCL-XL，MCL-1和akt下调。此外，尽管低浓度的硼替佐米不会导致细胞死亡，但17AAG和硼替佐米的组合治疗显示出对U266细胞系的协同凋亡作用。这些数据表明，靶向抑制HSP90可能被证明是开发MM患者未来治疗选择的有效且创新的策略。

BACKGROUND & AIMS: OBJECTIVE:Accumulating findings suggest that in acute myeloid leukemia (AML) patients, proinflammatory cytokines and growth factors play important roles in the proliferation and survival of AML cells in an autocrine and paracrine manner, leading to deterioration of AML. JTE-607 is a multiple cytokine inhibitor that potently suppresses production of proinflammatory cytokines. In the present study, we investigated the potency of JTE-607 as an antileukemic agent by exploiting a SCID mouse acute leukemia model. METHODS:SCID mice injected with anti-asialo-GM1 antibody were exposed to sublethal total-body irradiation at a dose of 3 Gy and then inoculated intravenously with AML cells. JTE-607 was administered using osmotic minipumps. The effects of JTE-607 on mouse survival time, human interleukin (IL)-8 levels in mouse plasma, and proportion of human CD45(+) cells in the bone marrow were studied. RESULTS:The survival time of the mice was strictly dependent on the number of U-937 cells proliferating in vivo. Administration of JTE-607 during the initial 7 days significantly prolonged survival of the mice, suggesting killing activity of JTE-607 against AML cells in vivo. Delayed administration of JTE-607 also prolonged the survival of mice bearing established leukemia with an effect comparable to the maximum tolerable dose of cytarabine. Flow cytometer analysis of bone marrow cells revealed decreased number of human CD45(+) cells. Human IL-8 level was also reduced by JTE-607. CONCLUSION:Our results indicate that JTE-607 has potential to be a new class of antileukemic drug that exerts inhibitory activities against both the proliferation and proinflammatory cytokine production of AML cells.

背景与目标：

BACKGROUND & AIMS: :Our previous studies have shown that aberrant arachidonic acid metabolism, especially the 5-lipoxygenase (5-Lox) pathway, is involved in oral carcinogenesis and can be targeted for cancer prevention. To develop potent topical agents for oral cancer chemoprevention, 5 known 5-Lox inhibitors from dietary and synthetic sources (Zileuton, ABT-761, licofelone, curcumin, and garcinol) were evaluated in silico for their potential efficacy. Garcinol, a polyisoprenylated benzophenone from the fruit rind of Garcinia spp., was found to be a promising agent based on the calculation of a theoretical activity index. Computer modeling showed that garcinol well fit the active site of 5-Lox, and potentially inhibited enzyme activity through interactions between the phenolic hydroxyl groups and the non-heme catalytic iron. In a short-term study on 7,12-dimethylbenz[a]anthracene (DMBA)-treated hamster cheek pouch, topical garcinol suppressed leukotriene B4 (LTB4) biosynthesis and inhibited inflammation and cell proliferation in the oral epithelium. In a long-term carcinogenesis study, topical garcinol significantly reduced the size of visible tumors, the number of cancer lesions, cell proliferation, and LTB4 biosynthesis. These results demonstrated that topical application of a 5-Lox inhibitor, garcinol, had chemopreventive effect on DMBA-induced hamster cheek pouch carcinogenesis.

背景与目标： : 我们以前的研究表明，异常的花生四烯酸代谢，尤其是5-脂氧合酶 (5-Lox) 途径，与口腔癌的发生有关，可以作为预防癌症的靶标。为了开发用于口腔癌化学预防的有效局部药物，在计算机上评估了饮食和合成来源的5种已知的5-Lox抑制剂 (Zileuton，ABT-761，licofelone，姜黄素和藤黄酚) 的潜在功效。根据理论活性指数的计算，发现藤黄酚是一种来自藤黄属果皮的聚异戊二烯化二苯甲酮，是一种有前途的试剂。计算机建模表明，藤酚非常适合5-Lox的活性位点，并通过酚羟基和非血红素催化铁之间的相互作用潜在地抑制了酶活性。在对7,12-二甲基苯并 [a] 蒽 (DMBA) 处理的仓鼠颊袋进行的短期研究中，外用藤黄酚抑制了白三烯B4 (LTB4) 的生物合成并抑制了口腔上皮的炎症和细胞增殖。在一项长期的致癌研究中，外用藤黄酚可显着减少可见肿瘤的大小，癌症病变的数量，细胞增殖和LTB4生物合成。这些结果表明，局部应用5-Lox抑制剂藤黄酚对DMBA诱导的仓鼠颊袋致癌具有化学预防作用。

BACKGROUND & AIMS: AIM:To evaluate photodynamic therapy (PDT) combined with the preferential the cyclooxygenase-2 (COX-2) inhibitor, nabumetone in the treatment of the neovascular age-related macular degeneration (ARMD). METHODS:A prospective, double-blind, randomized study on 60 patients with subfoveal CNV secondary to ARMD without any previous treatment. Patients were divided into a nabumetone or placebo group. The main endpoints were the change of best-corrected visual acuity (BCVA), central macular thickness (CRT) and number of required PDT treatments. RESULTS:In the nabumetone group, 27 patients (90%) and 28 (93%) in the placebo group completed the follow-up of 12 months. In the nabumetone group, the mean CRT decreased from 332 μm (SD 68 μm) to 220 μm (SD 46 μm). In the placebo group, CRT decreased from 331 μm (SD 72 μm) to 254 μm (SD 61 μm). The mean BCVA was 0.68 log MAR (SD 0.22 log MAR) in the nabumetone group and 0.62 log MAR (SD 0.23 log MAR) in the placebo group at baseline. This stabilised in the placebo group to 0.66 log MAR (SD 0.33) but deteriorated in the nabumetone group to 0.86 log MAR (SD 0.41 log MAR). There was a significant reduction in the number of required PDTs in the nabumetone group, but significant progression of the RPE atrophy area. CONCLUSION:Combined PDT with oral intake of the COX-2 inhibitor, nabumetone reduced the number of required PDT retreatments, but worsening BCVA caused by macular atrophy progression. Therefore the combination of the PDT with the nabumetone is not recommended.

背景与目标：

BACKGROUND & AIMS: :The ETV6-NTRK3 (EN) fusion gene which encodes a chimeric tyrosine kinase was first identified by cloning of the t(12;15)(p13;q25) translocation in congenital fibrosarcoma (CFS). Since then, EN has been also found in congenital mesoblastic nephroma (CMN), secretory breast carcinoma (SBC) and acute myelogenous leukemia (AML). Using IMS-M2 and M0-91 cell lines harboring the EN fusion gene, and Ba/F3 cells stably transfected with EN, we demonstrated that PKC412, also known as midostaurin, is an inhibitor of EN. Inhibition of EN activity by PKC412 suppressed the activity of it downstream molecules leading to inhibition of cell proliferation and induction of apoptosis. Our data for the first time suggested that PKC412 could serve as therapeutic drug for treatment of patients with this fusion.

背景与目标： : 首先通过克隆先天性纤维肉瘤 (CFS) 中的t(12;15)(p13;q25) 易位来鉴定编码嵌合酪氨酸激酶的ETV6-NTRK3 (EN) 融合基因。此后，在先天性中胚层肾瘤 (CMN)，分泌性乳腺癌 (SBC) 和急性骨髓性白血病 (AML) 中也发现了EN。使用带有EN融合基因的IMS-M2和M0-91细胞系以及用EN稳定转染的Ba/F3细胞，我们证明了PKC412 (也称为midostaurin) 是EN的抑制剂。PKC412对EN活性的抑制抑制了其下游分子的活性，从而抑制了细胞增殖并诱导了凋亡。我们的数据首次表明PKC412可以作为治疗这种融合患者的治疗药物。

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。利格列汀是临床抗2型糖尿病的二肽peptidase-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。为了了解疗效是否由血糖调节介导，与磺酰脲类格列美脲进行了比较。为了确定利格列汀介导的疗效是否取决于糖尿病背景，在非糖尿病小鼠中进行了实验。通过给小鼠喂食高脂饮食32周而诱发2型糖尿病。用利格列汀/格列美脲治疗小鼠7周。短暂性大脑中动脉闭塞在治疗4周时诱发中风。在整个实验中评估血液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。通过确定中风量和纹状体/皮质中存活神经元的立体定量来测量缺血性脑损伤。我们显示了利格列汀在2型糖尿病和正常小鼠中的明显抗中风功效，而格列美脲仅在正常小鼠中被证明对中风有效。这些结果表明利格列汀介导的神经保护作用是不依赖葡萄糖的并且可能涉及GLP-1。这些发现可能为糖尿病患者预防和治疗中风的DPP-4抑制剂的开发提供动力。

BACKGROUND & AIMS: :Fluoxetine (FLX) the active pharmaceutical ingredient (API) in Prozac(®) is a widely prescribed psychoactive drug which ubiquitous occurrence in the aquatic environment is associated to a poor removal rate in waste-water treatment plant (WWTP) systems. This API acts as a selective serotonin reuptake inhibitor (SSRI) frequently reported to cause disrupting effects in non-target species. The objective of this study includes a multibiomarker response evaluation on mussel Mytilus galloprovincialis during two weeks exposure to 75 ng L(-1) FLX assessing antioxidant enzymes activities--superoxide dismutase (SOD), catalase (CAT) and glutathione-S-transferase (GST); lipid peroxidation (LPO), acetylcholinesterase (AChE) neurotoxic response and endocrine disruption through alkali-labile phosphates (ALP) indirect measurement of vitellogenin-like proteins. Results show transient tissue-specific enzymatic responses and damage affecting mostly mussel gills. However, the clear ALP levels inhibition throughout time in both sex-differentiated gonads gives evidence to FLX reinforced action as an endocrine disruptor rather than an oxidative or neurotoxic inducer.

背景与目标： : 氟西汀 (FLX) 百忧解中的活性药物成分 (API) (®) 是一种广泛使用的精神活性药物，在水生环境中普遍存在，这与废水处理厂 (WWTP) 系统的去除率差有关。该API充当选择性5-羟色胺再摄取抑制剂 (SSRI)，经常报道会在非目标物种中引起破坏作用。这项研究的目的包括在暴露于75 ng L(-1) FLX的两周内对贻贝Mytilus galloprovincialis进行多生物标志物反应评估，评估抗氧化酶活性-超氧化物歧化酶 (SOD)，过氧化氢酶 (CAT) 和谷胱甘肽-S-转移酶 (GST); 脂质过氧化 (LPO)，乙酰胆碱酯酶 (AChE) 通过碱性不稳定磷酸盐 (ALP) 间接测量卵黄蛋白原样蛋白的神经毒性反应和内分泌破坏。结果显示，短暂的组织特异性酶反应和损伤主要影响贻贝g。然而，在两种性别分化的性腺中，整个过程中明显的ALP水平抑制提供了FLX作为内分泌干扰物而不是氧化或神经毒性诱导剂增强作用的证据。

BACKGROUND & AIMS: :The anti-inflammatory potential of p38 mitogen-activated protein kinase (MAPK) inhibitors was coincidentally expanded to a dual inhibition of p38α MAPK and phosphodiesterase 4 (PDE4), and the potential benefits arising from the blockage of both inflammation-related enzymes were thoroughly investigated. The most promising compound, CBS-3595 (1), was successively evaluated in in vitro experiments as well as in ex vivo and in vivo preclinical studies after administration of 1 to rodents, dogs, and monkeys. The resulting data clearly indicated a potent suppression of tumor necrosis factor alpha release. For reconfirming the findings of the animal studies when administering 1 to healthy human volunteers, a phase I clinical trial was conducted. Apart from further information regarding the pharmacokinetic and pharmacodynamic characteristics of 1, it was demonstrated that dual inhibition of p38α MAPK and PDE4 is able to synergistically attenuate the excessive anti-inflammatory response.

背景与目标： : p38丝裂原活化蛋白激酶 (MAPK) 抑制剂的抗炎潜力被巧合地扩展为对p38α MAPK和磷酸二酯酶4 (PDE4) 的双重抑制，并且两种炎症相关酶的阻断所产生的潜在益处被彻底研究。在对啮齿动物，狗和猴子施用1之后，在体外实验以及体外和体内临床前研究中相继评估了最有希望的化合物CBS-3595 (1)。所得数据清楚地表明有效抑制了肿瘤坏死因子 α 的释放。为了在向健康的人类志愿者施用1时再次确认动物研究的结果，进行了I期临床试验。除了有关1的药代动力学和药效学特征的进一步信息外，还证明了p38α MAPK和PDE4的双重抑制能够协同减弱过度的抗炎反应。

BACKGROUND & AIMS: PURPOSE:Many women with hormone receptor-positive breast cancer discontinue effective aromatase inhibitor (AI) treatment due to joint symptoms. METHODS:We conducted a single-arm, open-label, phase II study evaluating glucosamine-sulfate (1,500 mg/day) + chondroitin-sulfate (1,200 mg/day) for 24 weeks to treat joint pain/stiffness in postmenopausal women with early stage breast cancer who developed moderate-to-severe joint pain after initiating AIs. The primary endpoint was improvement in pain/stiffness at week 24 assessed by the Outcome Measure in Rheumatology Clinical Trials and Osteoarthritis Research Society International (OMERACT-OARSI) criteria. Secondary endpoints assessed changes in pain, stiffness, and function using the Western Ontario and McMaster Universities Osteoarthritis (WOMAC) Index for hips/knees and the Modified Score for the Assessment and Quantification of Chronic Rheumatoid Affections of the Hands (M-SACRAH) for hands/wrists. The Brief Pain Inventory (BPI) assessed pain interference, severity, and worst pain. RESULTS:Of 53 patients enrolled, 39 were evaluable at week 24. From baseline to week 24, 46 % of patients improved according to OMERACT-OARSI criteria. At week 24, there were improvements (all P < 0.05) in pain and function as assessed by WOMAC and M-SACRAH, and in pain interference, severity, and worst pain as assessed by BPI. Estradiol levels did not change from baseline. The most commonly reported side effects were headache (28 %), dyspepsia (15 %), and nausea (17 %). CONCLUSIONS:In this single-arm study, 24 weeks of glucosamine/chondroitin resulted in moderate improvements in AI-induced arthralgias, with minimal side effects, and no changes in estradiol levels. These results suggest a need to evaluate efficacy in a placebo-controlled trial.

背景与目标：

BACKGROUND & AIMS: :An extract from red ginseng [steamed and dried roots of Panax ginseng C.A. Meyer (RGE)] has been shown to have various actions on physiological functions. The mechanisms by which RGE promotes cholesterol metabolism in the liver are unclear, but RGE decreases the plasma levels of cholesterol. We investigated whether RGE affected the mRNA expression of cholesterol metabolism-related proteins such as cytochrome P450 (CYP)7A1 and bile salt export pump (BSEP) in the liver in hypercholesterolemic rats and rat primary hepatocytes. In-vivo studies showed the upregulation of CYP7A1 mRNA in hypercholesterolemic rats treated with RGE. Treatment with RGE exhibited decreased ratios of low-density lipoprotein-cholesterol to high-density lipoprotein-cholesterol compared with hypercholesterolemia without RGE. In-vitro studies also showed the upregulation of CYP7A1 mRNA and protein levels by the addition of RGE to rat primary hepatocytes. The mRNA levels of BSEP exhibited few changes. The sustained levels of the liver X receptor (LXR) in vivo and the increased levels of LXR in vitro on RGE treatment could be involved in the upregulation of CYP7A1. To clarify the effects of 11 ginsenosides including RGE on the mRNA levels of CYP7A1 and BSEP, we performed in-vitro experiments using rat primary hepatocytes. The ginsenosides Ro, Rg3, Re, Rg1, and Rg2 exhibited increased mRNA levels of CYP7A1. These results suggest that several ginsenosides including RGE promoted cholesterol metabolism due to upregulation of CYP7A1.

背景与目标： : 来自红参的提取物 [人参c.A.的蒸干根Meyer (RGE)] 已被证明对生理功能具有多种作用。RGE促进肝脏中胆固醇代谢的机制尚不清楚，但RGE会降低血浆胆固醇水平。我们研究了RGE是否影响高胆固醇血症大鼠和大鼠原代肝细胞肝脏中胆固醇代谢相关蛋白 (例如细胞色素P450 (CYP)7A1和胆盐出口泵 (BSEP)) 的mRNA表达。体内研究表明，用RGE治疗的高胆固醇血症大鼠CYP7A1 mRNA上调。与没有RGE的高胆固醇血症相比，RGE治疗显示低密度脂蛋白胆固醇与高密度脂蛋白胆固醇的比率降低。体外研究还显示，通过向大鼠原代肝细胞添加RGE，CYP7A1 mRNA和蛋白质水平上调。BSEP的mRNA水平几乎没有变化。RGE治疗后体内肝脏X受体 (LXR) 的持续水平和体外LXR水平的升高可能与CYP7A1的上调有关。为了阐明包括RGE在内的11种人参皂甙对CYP7A1和BSEP mRNA水平的影响，我们使用大鼠原代肝细胞进行了体外实验。人参皂苷Ro，Rg3，Re，Rg1和Rg2显示CYP7A1的mRNA水平升高。这些结果表明，由于CYP7A1的上调，包括RGE在内的几种人参皂甙可促进胆固醇代谢。

BACKGROUND & AIMS: :Plasminogen activator inhibitor-1 (PAI-1) is a member of the serine protease inhibitor (serpin) superfamily, which inactivates tissue plasminogen activator (tPA); therefore, increased level of PAI-1 antigen counteracts the anticoagulant effect of tPA and facilitates the fibrin clot formation. Plasma PAI-1 antigen and activity levels are associated with increased body mass index and with features of the insulin resistance syndrome like obesity and diabetes. Visceral adipose tissue produces more PAI-1 than subcutaneous adipose tissue: This increased production of PAI-1 from the visceral adipose tissue is one important link between visceral obesity and cardiovascular disease. Besides visceral adipose tissue, there is mounting evidence that epicardial adipose tissue may be an important source of PAI-1, especially in patients with type 2 diabetes.

背景与目标： : 纤溶酶原激活物抑制剂-1 (PAI-1) 是丝氨酸蛋白酶抑制剂 (serpin) 超家族的成员，其使组织纤溶酶原激活物 (tPA) 失活; 因此，PAI-1抗原水平的升高抵消了tPA的抗凝作用并促进了纤维蛋白凝块的形成。血浆PAI-1抗原和活性水平与体重指数增加以及胰岛素抵抗综合征 (如肥胖和糖尿病) 的特征相关。内脏脂肪组织比皮下脂肪组织产生更多的PAI-1: 内脏脂肪组织产生的PAI-1增加是内脏肥胖和心血管疾病之间的重要联系。除了内脏脂肪组织外，越来越多的证据表明心外膜脂肪组织可能是PAI-1的重要来源，尤其是在2型糖尿病患者中。

BACKGROUND & AIMS: Prostaglandins and thromboxanes are products of arachidonic acid metabolism via the cyclooxygenase (CO) enzyme and are responsible for the pain and swelling common to sites of inflammation. Non-steroidal anti-inflammatory drugs (NSAIDs) inhibit the production of these substances and are used in the treatment of inflammatory diseases such as arthritis. However, one of the major side-effects of NSAID therapy is gastric ulceration. It is possible that inhibition of prostaglandin production and a related increase in the formation of leukotrienes via the 5-lipoxygenase (5-LO) enzymatic pathway are responsible for attracting inflammatory cells, causing local sites of inflammation and producing ulceration. To determine the effects of 5-LO inhibition on this hypothesis, studies were performed in rats to evaluate the effects of tepoxalin, a dual CO/LO inhibitor on leukotriene B4 levels in gastric mucosa and neutrophil adhesion in mesenteric venules. In rats, chronic oral administration of an NSAID, indomethacin (2 mg/kg daily over 4 days), resulted in 40% mortality, accompanied by intestinal adhesions and perforations when evaluated 24 h after the fourth dose of drug. Additionally, neutrophil adhesion was increased in the mesenteric venules and cell infiltration was evident in the mesenteric interstitium. These gastrointestinal side-effects were inhibited in a separate group of rats administered tepoxalin (20 mg/kg, p.o) 30 min prior to each daily indomethacin treatment. Further studies were performed to determine tepoxalin's effects on early events associated with NSAID-induced gastrointestinal inflammation, including neutrophil adhesion, lipid peroxide generation and LTB4 production. Indomethacin (100 mg/kg, p.o.) produced elevated levels of LTB4 in rat gastric mucosa 90 min after administration. Additionally, neutrophil adhesion in mesenteric venules was increased at this dose and with the administration of another NSAID, naproxen. No generation of lipid peroxides was evident in the gastric mucosa at this timepoint. Tepoxalin (up to 400 mg/kg, p.o.) did not have an effects on gastric mucosal LTB4 generation and lipid peroxide levels. A decrease in neutrophil adhesion was observed at the highest dose. In another study, pretreatment with tepoxalin (ED50=7.5 mg/kg, p.o.) or the selective 5-LO inhibitor zileuton (100 mg/kg, p.o.) prevented the increases in gastric mucosal LTB4 levels and neutrophil adhesion induced by indomethacin (100 mg/kg, p.o.). These data suggest that LO inhibition may play a vital role in the prevention of NSAID-induced gastric inflammation, providing insight into the lack of ulcerogenicity with tepoxalin and new approaches to anti-inflammatory therapy which may prevent gastric side effects.

背景与目标： 前列腺素和血栓烷是花生四烯酸通过环氧合酶 (CO) 酶代谢的产物，并负责炎症部位常见的疼痛和肿胀。非甾体抗炎药 (NSAIDs) 抑制这些物质的产生，并用于治疗炎症性疾病，例如关节炎。然而，NSAID治疗的主要副作用之一是胃溃疡。通过5-脂氧合酶 (5-LO) 酶途径抑制前列腺素的产生和白三烯形成的相关增加可能导致吸引炎症细胞，引起局部炎症部位并产生溃疡。为了确定5-LO抑制对该假设的影响，在大鼠中进行了研究，以评估tepoxalin (一种双重CO/LO抑制剂) 对胃粘膜中白三烯B4水平和肠系膜小静脉中性粒细胞粘附的影响。在大鼠中，长期口服NSAID吲哚美辛 (在4天内每天2 mg/kg) 导致40% 死亡率，并在第四剂药物后24小时评估时伴有肠粘连和穿孔。此外，肠系膜小静脉中的中性粒细胞粘附增加，肠系膜间质中的细胞浸润明显。在每天吲哚美辛治疗前30分钟，在单独的大鼠组中抑制了这些胃肠道副作用。进行了进一步的研究以确定tepoxalin对与NSAID诱导的胃肠道炎症相关的早期事件的影响，包括中性粒细胞粘附，脂质过氧化物生成和LTB4生成。给药90分钟后，吲哚美辛 (100 mg/kg，p.o.) 在大鼠胃粘膜中产生升高的LTB4水平。此外，在该剂量下以及使用另一种NSAID萘普生时，肠系膜小静脉中的中性粒细胞粘附增加。此时，胃粘膜中没有明显的脂质过氧化物生成。Tepoxalin (最高400 mg/kg，p.o.) 对胃粘膜LTB4的生成和脂质过氧化物水平没有影响。在最高剂量下观察到中性粒细胞粘附减少。在另一项研究中，用tepoxalin (ED50 = 7.5 mg/kg，p.o.) 或选择性5-LO抑制剂zileuton (100 mg/kg，p.o.) 进行预处理可防止胃粘膜LTB4水平升高和中性粒细胞粘附由吲哚美辛 (100 mg/kg，p、o.)。这些数据表明，LO抑制作用可能在预防NSAID诱导的胃部炎症中起着至关重要的作用，从而深入了解tepoxalin缺乏溃疡性以及可能预防胃副作用的抗炎治疗的新方法。

BACKGROUND & AIMS: 1. Selective 5-hydroxytryptamine (5-HT; serotonin) reuptake inhibitors (SSRIs) cause a greater increase in extracellular 5-HT in the forebrain when the somatodendritic 5-HT1A autoreceptor is blocked. Here, we investigated whether blockade of the terminal 5-HT1B autoreceptor influences a selective 5-HT reuptake inhibitor in the same way, and whether there is an additional effect of blocking both the 5-HT1A and 5-HT1B autoreceptors. 2. Extracellular 5-HT was measured in frontal cortex of the anaesthetized rat by use of brain microdialysis. In vivo extracellular recordings of 5-HT neuronal activity in the dorsal raphe nucleus (DRN) were also carried out. 3. The selective 5-HT reuptake inhibitor, paroxetine (0.8 mg kg-1, i.v.), increased extracellular 5-HT about 2 fold in rats pretreated with the 5-HT1A receptor antagonist, WAY100635. When administered alone neither paroxetine (0.8 mg kg-1, i.v.) nor WAY100635 (0.1 mg kg-1, i.v.) altered extracellular 5-HT levels. 4. Paroxetine (0.8 mg kg-1, i.v.) did not increase 5-HT in rats pretreated with the 5-HT1B/D receptor antagonist, GR127935 (1 mg kg-1, i.v.). GR127935 (1 and 5 mg kg-1, i.v.) had no effect on extracellular 5-HT when administered alone. 5. Interestingly, paroxetine (0.8 mg kg-1, i.v.) caused the greatest increase in 5-HT (up to 5 fold) when GR127935 (1 or 5 mg kg-1, i.v.) was administered in combination with WAY100635 (0.1 mg kg-1, i.v.). Administration of GR127935 (5 mg kg-1, i.v.) plus WAY100635 (0.1 mg kg-1, i.v.) without paroxetine, had no effect on extracellular 5-HT in the frontal cortex. 6. Despite the lack of effect of GR127935 on 5-HT under basal conditions, when 5-HT output was elevated about 3 fold (by adding 1 microM paroxetine to the perfusion medium), the drug caused a dose-related (1 and 5 mg kg-1, i.v.) increase in 5-HT. 7. By itself, GR127935 slightly but significantly decreased 5-HT cell firing in the DRN at higher doses (2.0-5.0 mg kg-1, i.v.), but did not prevent the inhibition of 5-HT cell firing induced by paroxetine. 8. In summary, our results suggest that selective 5-HT reuptake inhibitors may cause a large increase in 5-HT in the frontal cortex when 5-HT autoreceptors on both the somatodendrites (5-HT1A) and nerve terminals (5-HT1B) are blocked. This increase is greater than when either set of autoreceptors are blocked separately. The failure of a 5-HT1B receptor antagonist alone to enhance the effect of the selective 5-HT reuptake inhibitor in our experiments may be related to a lack of tone on the terminal 5-HT1B autoreceptor due to a continued inhibition of 5-HT cell firing. These results are discussed in relation to the use of 5-HT autoreceptor antagonists to augment the antidepressant effect of selective 5-HT reuptake inhibitors.

背景与目标： 1.选择性5-羟色胺 (5-HT; 5-羟色胺) 再摄取抑制剂 (SSRIs) 在体细胞肌腱5-HT1A自身受体受阻时，会导致前脑细胞外5-HT的增加。在这里，我们研究了末端5-HT1B自身受体的阻断是否以相同的方式影响选择性5-HT再摄取抑制剂，以及是否存在阻断5-HT1A和5-HT1B自身受体的附加作用。2.使用脑微透析在麻醉的大鼠额叶皮层中测量细胞外5-HT。还进行了中缝背核 (DRN) 中5-HT神经元活性的体内细胞外记录。3.在用5-HT1A受体拮抗剂way100635预处理的大鼠中，选择性5-HT再摄取抑制剂帕罗西汀 (0.8 mg kg-1，i.v.) 使细胞外5-HT增加约2倍。当单独给药时，帕罗西汀 (0.8 mg kg-1，静脉注射) 和WAY100635 (0.1 mg kg-1，静脉注射) 都不会改变细胞外5-HT水平。4.在用5-HT1B/D受体拮抗剂GR127935 (1 mg kg-1，i.v.) 预处理的大鼠中，帕罗西汀 (0.8 mg kg-1，i.v.) 不增加5-HT。GR127935 (1和5 mg kg-1，静脉注射) 单独给药时对细胞外5-HT没有影响。5.有趣的是，当GR127935 (1或5 mg kg-1，静脉注射) 与WAY100635 (0.1 mg kg-1，静脉注射) 联合给药时，帕罗西汀 (0.8 mg kg-1，静脉注射) 引起5-HT的最大增加 (高达5倍)。i.v.)。施用GR127935 (5 mg kg-1，i.v.) 加WAY100635 (0.1 mg kg-1，i.v.) 而不使用帕罗西汀，对额叶皮层的细胞外5-HT没有影响。6.尽管在基础条件下GR127935对5-HT缺乏作用，但当5-HT输出量增加约3倍 (通过向灌注培养基中添加1 microM帕罗西汀) 时，药物导致5-HT剂量相关 (1和5 mg kg-1，静脉注射) 增加。7.就其本身而言，GR127935在较高剂量 (2.0-5.0 mg kg-1，i.v.) 下略微但显著降低DRN中的5-HT细胞放电，但没有阻止帕罗西汀诱导的5-HT细胞放电的抑制。8.总之，我们的结果表明，选择性5-HT再摄取抑制剂可能会导致额叶皮层5-HT大量增加，当体突肌 (5-HT1A) 和神经末梢 (5-HT1B) 上的5-HT自身受体被阻断。此增加大于单独阻塞任一组自身感受器时的增加。在我们的实验中，单独的5-HT1B受体拮抗剂未能增强选择性5-HT再摄取抑制剂的作用，这可能与由于持续抑制5-HT1B自身受体而导致末端5-HT1B自身受体缺乏音调有关。ht细胞放电。讨论了这些结果，这些结果与使用5-HT自身受体拮抗剂来增强选择性5-HT再摄取抑制剂的抗抑郁作用有关。

BACKGROUND & AIMS: :Direct thrombin inhibitors have proven efficacious in prevention of venous thromboembolism. Bleeding complications are rare, but in case of acute serious bleeding, an effective and instant haemostatic intervention may be required. In the present study it was demonstrated that the direct thrombin inhibitor melagatran induces dose-dependent abnormalities in whole blood (WB) clotting profiles as recorded by a recently described modified thrombelastographic model, and that rFVIIa or APCC are capable of improving the haemostatic capacity. Experiments were performed using WB from 30 healthy males. In-vitro titration experiments (n = 10) with addition of melagatran to WB corresponding to plasma concentrations ranging from 0 to 5.0 microM (12 steps) showed a dose-dependent prolongation of the clot initiation and characteristic decrease of the maximum rate of clot propagation. In-vitro intervention studies (n = 20) were completed with four different concentrations of melagatran as well as addition of four different levels of rFVIIa or APCC. At all tested concentrations of melagatran, rFVIIa significantly shortened the melagatran-induced prolonged clot initiation but induced only minor improvements of the reduced clot propagation. In contrast, APCC significantly and dose-dependently shortened the clot initiation and accelerated the clot propagation. In conclusion, our thrombelastographic model appears useful for evaluating the effect of direct thrombin inhibitors on dynamicWB clot formation and rFVIIa, but especially APCC significantly improved theWB clot formation. The pronounced stabilizing effect of APCC may be caused by its content of prothrombin and activated coagulation factors.

背景与目标： : 直接凝血酶抑制剂已被证明对预防静脉血栓栓塞有效。出血并发症很少见，但在急性严重出血的情况下，可能需要有效且即时的止血干预。在本研究中，证明了直接凝血酶抑制剂melagatran诱导全血 (WB) 凝血谱的剂量依赖性异常，如最近描述的改良血栓弹力图模型所记录，并且rFVIIa或APCC能够改善止血能力。使用来自30名健康男性的WB进行了实验。将melagatran添加到WB中的体外滴定实验 (n = 10)，对应于血浆浓度范围为0至5.0微米 (12步)，结果显示凝块起始的剂量依赖性延长和最大凝块传播速率的特征性降低。用四种不同浓度的美拉加群以及添加四种不同水平的rFVIIa或APCC完成了体外干预研究 (n = 20)。在所有测试的melagatran浓度下，rFVIIa均显着缩短了melagatran诱导的长时间凝块起始，但仅诱导了减少的凝块繁殖的微小改善。相反，APCC显着且剂量依赖性地缩短了凝块的起始并加速了凝块的传播。总之，我们的血栓弹力图模型似乎可用于评估直接凝血酶抑制剂对动态wb凝块形成和rFVIIa的影响，但尤其是APCC显着改善了wb凝块形成。APCC的明显稳定作用可能是由其凝血酶原和活化的凝血因子的含量引起的。

BACKGROUND & AIMS: :Plasmepsins are aspartic proteases involved in the degradation of the host cell hemoglobin that is used as a food source by the malaria parasite. Plasmepsins are highly promising as drug targets, especially when combined with the inhibition of falcipains that are also involved in hemoglobin catabolism. In this review, we discuss the mechanism of plasmepsins I-IV in view of the interest in transition state mimetics as potential compounds for lead development. Inhibitor development against plasmepsin II as well as relevant crystal structures are summarized in order to give an overview of the field. Application of computational techniques, especially binding affinity prediction by the linear interaction energy method, in the development of malarial plasmepsin inhibitors has been highly successful and is discussed in detail. Homology modeling and molecular docking have been useful in the current inhibitor design project, and the combination of such methods with binding free energy calculations is analyzed.

背景与目标： : 血浆蛋白酶是参与宿主细胞血红蛋白降解的天冬氨酸蛋白酶，被疟原虫用作食物来源。血浆蛋白酶作为药物靶标非常有希望，尤其是与抑制也参与血红蛋白分解代谢的falcipains结合使用时。在这篇综述中，鉴于过渡态模拟物作为潜在的铅开发化合物的兴趣，我们讨论了血浆蛋白酶i-iv的机理。总结了针对plasmepsin II的抑制剂开发以及相关的晶体结构，以概述该领域。计算技术的应用，尤其是通过线性相互作用能方法进行的结合亲和力预测，在疟疾血浆蛋白酶抑制剂的开发中非常成功，并进行了详细讨论。同源性建模和分子对接在当前的抑制剂设计项目中很有用，并且分析了此类方法与结合自由能计算的结合。