Everolimus combined with cisplatin has a potential role in treatment of urothelial bladder cancer

Cisplatin (CDDP)-based chemotherapy is a commonly treatment for advanced urothelial carcinoma. However, episodes of cisplatin resistance have been referenced. Recently it has been reported that everolimus (RAD001) could have an important role to play in bladder-cancer treatment and that mTOR inhibitors may restore chemosensitivity in resistant tumours. The aim of this study was to assess RAD001 in vitro ability to enhance CDDP cytotoxicity in three human bladder-cancer cell lines. Over the course of 72 h, the cells were exposed to different concentrations of CDDP and RAD001, isolated or combined. Treatment with CDDP statistically (P < 0.05) decreased cell proliferation in cell lines in a dose-dependent manner. The anti-proliferative activity of CDDP used in combination with RAD001 was statistically significant (P < 0.05) in the cell lines at all concentrations tested. RAD001 had a therapeutic effect when used in combination with CDDP and could therefore be a useful anti-cancer drug combination for patients with bladder cancer.     

Detection of T-regulatory cells has a potential role in the diagnosis of classical Hodgkin lymphoma

Previous studies have demonstrated an increase in T-regulatory cells in the involved lymph nodes and peripheral blood of patients with Hodgkin lymphoma. Our study examined whether the detection of T-regulatory cells by flow cytometry could distinguish classical Hodgkin lymphoma (CHL) from benign cases and B-cell non-Hodgkin lymphomas (B-NHL). We measured CD4, CD25, and CD152 in 14 CHLs, 2 nodular lymphocyte-predominant Hodgkin lymphomas, 31 B-NHLs, and 54 benign cases. All T-regulatory cell parameters, including percent lymphocytes CD4+/CD152+ and CD4+/CD25+/CD152+, and mean and median CD152 expression in CD4+/CD25+ lymphocytes, were higher in CHL than in B-NHL and benign. Mean CD152 in CD4+/CD25+ lymphocytes distinguished CHL from benign with 79% sensitivity and 100% specificity, and from B-NHL with 71% sensitivity and 90% specificity. Overall, our results show that T-regulatory cells are increased in CHL and their detection may be a useful tool in differentiating CHL from other entities.     

Interaction of vasopressins and linear N-terminal ACTH fragments in the induction of natriuresis

Abstract. Because a) a mixture of alpha-melanotropin (MSH) and corticotropin (1–13)-tridecapeptide (1–13 ACTH) has apparently been isolated from natriuretic fractions of posterior pituitary which also contained arginine-vasopressin, and b) the natriuretic activity of the isolated fraction was greater than that of any of the above three peptides (synthetic) given separately, possible interaction was studied with model substances in the chloralosed cat preparation. It was found that with both alpha-MSH and lysine-vasopressin, extracted samples were more natriuretic than synthetic samples despite an equivalence of pressor activities. When combined with arginine-vasopressin in one injectate, synthetic alpha-MSH, 1–4 ACTH, 1–13 ACTH-amide and 1–18 ACTH all showed potentiation of natriuretic activities with no effect on pressor activity. This effect was not shown by synthetic 4–10 ACTH and 5–18 ACTH, or by extracted alpha-MSH. It was concluded that the 1–4 sequence Ser-Tyr-Ser-Met, regardless of whether the N-terminal is free or blocked, can potentiate the natriuretic action of vasopressin.     

The FGF system has a key role in regulating vascular integrity

The integrity of the endothelial monolayer is essential to blood vessel homeostasis and active regulation of endothelial permeability. The FGF system plays important roles in a wide variety of physiologic and pathologic conditions; however, its role in the adult vasculature has not been defined. To assess the role of the FGF system in the adult endothelial monolayer, we disrupted FGF signaling in bovine aortic endothelial cells and human saphenous vein endothelial cells in vitro and in adult mouse and rat endothelial cells in vivo using soluble FGF traps or a dominant inhibitor of all FGF receptors. The inhibition of FGF signaling using these approaches resulted in dissociation of the VE-cadherin/p120-catenin complex and disassembly of adherens and tight junctions, which progressed to loss of endothelial cells, severe impairment of the endothelial barrier function, and finally, disintegration of the vasculature. Thus, FGF signaling plays a key role in the maintenance of vascular integrity.     

阿托品对四氧嘧啶糖尿病动物模型的影响

目的：探讨四氧嘧啶配伍阿托品对制作糖尿病动物模型的影响。 方法：实验于2005-07／09在广西医科大学药学院药理实验室完成。选取小鼠40只禁食12h，随机分为对照组、四氧嘧啶组（50mg／kg腹腔注射&;#215;2）、四氧嘧啶配伍高剂量（2mg／kg腹腔注射&;#215;4）阿托品组、四氧嘧啶配伍中剂量（1mg／kg腹腔注射&;#215;4）阿托品组，各10只。配伍阿托品组分别于注射四氧嘧啶前0．5h，后0．5h，2，3d共4次给予腹腔注射阿托品，全部在造模后4，14d测定空腹血糖。选取大鼠58只禁食20h，随机分为高剂量（130mg／kg腹腔注射&;#215;2）四氧嘧啶组、中剂量（100mg／kg腹腔注射&;#215;2）四氧嘧啶组、高剂量四氧嘧啶配伍阿托品组、中剂量四氧嘧啶配伍阿托品组和对照组，各组分别为14，10，14，10，10只。除对照组注射等量生理盐水外，其余各组均注射四氧嘧啶，分别于注射四氧嘧啶前0．5h，后0．5h，2，3d共4次给予腹腔注射阿托品（2mg／kg），并于造模后4，14，30d测定空腹血糖，以空腹血糖≥11．1mmol／L判定为糖尿病模型。测定各组动物的空腹血糖及血糖缓解幅度【血糖缓解幅度（％）=（前高血糖数值-后高血糖数值）／前高血糖数值&;#215;100％】，计算糖尿病大鼠成模率、累计死亡率、平均存活时间。 结果：纳人小鼠40只和大鼠58只，38只小鼠和51只大鼠进人结果分析。其中造模第2天四氧嘧啶组小鼠死亡1只，采血时四氧嘧啶配伍中剂量阿托品组小鼠死亡1只；采血测血糖前高剂量四氧嘧啶组和高剂量四氧嘧啶配伍阿托品组大鼠分别累计死亡2只和3只。采血时此两组又各死亡1只。①小鼠四氧嘧啶配伍高剂量和中剂量阿托品组高血糖缓解幅度均较单用四氧嘧啶组明显（分别为10．3％，10．6％，47．5％，P〈0．01），高剂量阿托品血糖稳定效果更佳。②四氧嘧啶配伍高剂量阿托品，造模后4d大鼠血糖水平升幅较四氧嘧啶组小[四氧嘧啶配伍高剂量阿托品组、四氧嘧啶组分别为（11．6&;#177;3．6），（21．0&;#177;6．7）mmol／L]，造模后30d血糖稳步升高，表现出缓慢温和及滞后特点，且高水平血糖的稳定性较好[四氧嘧啶配伍高剂量阿托品组、四氧嘧啶组分别为（15．3&;#177;5．9），（13．3&;#177;5．5）mmol／L]。③与同量四氧嘧啶组比较，造模后30d大鼠中剂量四氧嘧啶配伍高剂量阿托品者成模率高（86％，80％）而死亡率低（30％，50％），平均存活时间延长（22A，15．8d）；造模后30d高剂量四氧嘧啶配伍高剂量阿托品者成模率高（75％，67％）、死亡率也高（71％，57％），平均存活时间较短（7．7，8．3d）。④胰腺病理显示，四氧嘧啶配伍阿托品者胰岛稀少，胰岛萎缩，β细胞破坏严重，而单用四氧嘧啶缓解者胰岛受损较轻，可见胰岛细胞修复增生。 结论：中剂量四氧嘧啶配伍高剂量阿托品制作糖尿病动物模型效果较好，能提高糖尿病成模率、降低死亡率并使高血糖水平较为稳定。     

阿托品对四氧嘧啶糖尿病动物模型的影响

目的:探讨四氧嘧啶配伍阿托品对制作糖尿病动物模型的影响。方法:实验于2005-07/09在广西医科大学药学院药理实验室完成。选取小鼠40只禁食12h,随机分为对照组、四氧嘧啶组(50mg/kg腹腔注射×2)、四氧嘧啶配伍高剂量(2mg/kg腹腔注射×4)阿托品组、四氧嘧啶配伍中剂量(1mg/kg腹腔注射×4)阿托品组,各10只。配伍阿托品组分别于注射四氧嘧啶前0.5h,后0.5h,2,3d共4次给予腹腔注射阿托品,全部在造模后4,14d测定空腹血糖。选取大鼠58只禁食20h,随机分为高剂量(130mg/kg腹腔注射×2)四氧嘧啶组、中剂量(100mg/kg腹腔注射×2)四氧嘧啶组、高剂量四氧嘧啶配伍阿托品组、中剂量四氧嘧啶配伍阿托品组和对照组,各组分别为14,10,14,10,10只。除对照组注射等量生理盐水外,其余各组均注射四氧嘧啶,分别于注射四氧嘧啶前0.5h,后0.5h,2,3d共4次给予腹腔注射阿托品(2mg/kg),并于造模后4,14,30d测定空腹血糖,以空腹血糖≥11.1mmol/L判定为糖尿病模型。测定各组动物的空腹血糖及血糖缓解幅度[血糖缓解幅度(%)=(前高血糖数值-后高血糖数值)/前高血糖数值×100%],计算糖尿病大鼠成模率、累计死亡率、平均存活时间。结果:纳入小鼠40只和大鼠58只,38只小鼠和51只大鼠进入结果分析。其中造模第2天四氧嘧啶组小鼠死亡1只,采血时四氧嘧啶配伍中剂量阿托品组小鼠死亡1只;采血测血糖前高剂量四氧嘧啶组和高剂量四氧嘧啶配伍阿托品组大鼠分别累计死亡2只和3只,采血时此两组又各死亡1只。①小鼠四氧嘧啶配伍高剂量和中剂量阿托品组高血糖缓解幅度均较单用四氧嘧啶组明显(分别为10.3%,10.6%,47.5%,P<0.01),高剂量阿托品血糖稳定效果更佳。②四氧嘧啶配伍高剂量阿托品,造模后4d大鼠血糖水平升幅较四氧嘧啶组小[四氧嘧啶配伍高剂量阿托品组、四氧嘧啶组分别为(11.6±3.6),(21.0±6.7)mmol/L],造模后30d血糖稳步升高,表现出缓慢温和及滞后特点,且高水平血糖的稳定性较好[四氧嘧啶配伍高剂量阿托品组、四氧嘧啶组分别为(15.3±5.9),(13.3±5.5)mmol/L]。③与同量四氧嘧啶组比较,造模后30d大鼠中剂量四氧嘧啶配伍高剂量阿托品者成模率高(86%,80%)而死亡率低(30%,50%),平均存活时间延长(22.4,15.8d);造模后30d高剂量四氧嘧啶配伍高剂量阿托品者成模率高(75%,67%)、死亡率也高(71%,57%),平均存活时间较短(7.7,8.3d)。④胰腺病理显示,四氧嘧啶配伍阿托品者胰岛稀少,胰岛萎缩,β细胞破坏严重,而单用四氧嘧啶缓解者胰岛受损较轻,可见胰岛细胞修复增生。结论:中剂量四氧嘧啶配伍高剂量阿托品制作糖尿病动物模型效果较好,能提高糖尿病成模率、降低死亡率并使高血糖水平较为稳定。     

PD-1/PD-L1在脓毒症免疫抑制中的潜在作用

脓毒症是宿主对炎症反应失调引起的危及生命的器官功能障碍[1].免疫抑制在脓毒症发展过程中发挥重要作用,它是导致脓毒症患者二次感染及病死率居高不下的重要原因[2],其主要表现为免疫细胞活性降低、数量减少.程序性细胞死亡受体-l(programmed cell deathprotein 1,PD-1)/程序性细胞死亡配体-1(programmed cell death-ligand 1,PD-L1)通过负向调节免疫细胞,从而引起免疫抑制,对脓毒症患者免疫功能及预后产生重要影响.本文就脓毒症免疫抑制机制及PD-1/PD-L1在其中的潜在作用做一简要综述.     

Evidence for a humoral mechanism in volume expansion natriuresis

The role of a humoral mechanism in the natriuresis induced by volume expansion was evaluated using an isolated dog kidney perfused by a second dog which had been pretreated with desoxycorticosterone acetate (DOCA). Expansion of the perfusion dog with an equilibrated volume of blood from a reservoir, resulted in an increase in UnaV (sodium excretion) from 153.6+/-27.9 (sem) to 345.5+/-57.8 muEq/min, P<0.001. FEna (fractional sodium excretion) increased from 3.4+/-0.6 to 8.1+/-1.2%, P<0.01. The natriuresis occurred in the face of a significant decrease in Cin, RBF, and renal arterial pressure, and in the absence of any change in plasma protein concentration or packed cell volume. In a control group of experiments, sodium excretion did not change when the perfusion dog was not volume expanded, although Cin (inulin clearance) and RBF (renal blood flow) decreased to the same degree as in the expanded group. These data support the conclusion that volume expansion of the perfusion dog either stimulated the release of a natriuretic factor or suppressed the release of an antinatriuretic factor which was manifested by an increase in sodium excretion in the isolated kidney.     

An internal polyadenylation signal substantially increases expression levels of lentivirus-delivered transgenes but has the potential to reduce viral titer in a promoter-dependent manner

In lentiviral gene delivery systems, transgene expression cassettes are commonly cloned without a polyadenylation signal to prevent disruption of full-length lentiviral genomes on mRNA maturation in producer cells. The lack of the polyadenylation signal, however, has the potential to reduce stability and translation efficiency of transgene mRNA. Therefore, we have assessed the effect of a strong internal polyadenylation [poly(A)] signal on both transgene expression levels in virus-infected cells and functional viral titer, in a series of eight self-inactivating lentiviruses expressing the mOrange transgene under the control of the constitutive cytomegalovirus (CMV), elongation factor 1alpha (EF1alpha), and beta-actin promoters or the highly tissue-specific prostate-specific antigen/probasin hybrid (PSA/Pb) promoter with or without a simian virus 40 (SV40) early polyadenylation signal downstream of the mOrange-coding sequence. We show that mOrange expression levels in virus-infected HEK-293, LNCaP, and primary prostate epithelial cells were increased 3- to 6.5-fold when an internal polyadenylation signal was present. When the CMV and EF1alpha promoters were used, functional viral titer decreased 8- to 9-fold in the presence of the polyadenylation signal, but titer was not affected when transgene expression was driven by the beta-actin promoter or tissue-specific PSA/Pb promoter. We therefore conclude that an internal polyadenylation signal in lentiviral vectors has a highly beneficial effect on transgene expression, but reduces viral titer in a promoter-dependent manner.     

Histamine-releasing factor has a proinflammatory role in mouse models of asthma and allergy

IgE-mediated activation of mast cells and basophils underlies allergic diseases such as asthma. Histamine-releasing factor (HRF; also known as translationally controlled tumor protein [TCTP] and fortilin) has been implicated in late-phase allergic reactions (LPRs) and chronic allergic inflammation, but its functions during asthma are not well understood. Here, we identified a subset of IgE and IgG antibodies as HRF-interacting molecules in vitro. HRF was able to dimerize and bind to Igs via interactions of its N-terminal and internal regions with the Fab region of Igs. Therefore, HRF together with HRF-reactive IgE was able to activate mast cells in vitro. In mouse models of asthma and allergy, Ig-interacting HRF peptides that were shown to block HRF/Ig interactions in vitro inhibited IgE/HRF-induced mast cell activation and in vivo cutaneous anaphylaxis and airway inflammation. Intranasally administered HRF recruited inflammatory immune cells to the lung in naive mice in a mast cell- and Fc receptor-dependent manner. These results indicate that HRF has a proinflammatory role in asthma and skin immediate hypersensitivity, leading us to suggest HRF as a potential therapeutic target.     

The role of lipid droplets in metabolic disease in rodents and humans

Lipid droplets (LDs) are intracellular organelles that store neutral lipids within cells. Over the last two decades there has been a dramatic growth in our understanding of LD biology and, in parallel, our understanding of the role of LDs in health and disease. In its simplest form, the LD regulates the storage and hydrolysis of neutral lipids, including triacylglycerol and/or cholesterol esters. It is becoming increasingly evident that alterations in the regulation of LD physiology and metabolism influence the risk of developing metabolic diseases such as diabetes. In this review we provide an update on the role of LD-associated proteins and LDs in metabolic disease.     

Dural fibroblasts play a potential role in headache pathophysiology

Nociceptive signaling from the meninges is proposed to contribute to many forms of headache. However, the events within the meninges that drive afferent activity are not clear. Meningeal fibroblasts are traditionally thought to produce extracellular proteins that constitute the meninges but not to contribute to headache. The purpose of these studies was to determine whether dural fibroblasts release factors that activate/sensitize dural afferents and produce headache-like behavior in rats. Dura mater was removed from male rats and dural fibroblasts were cultured. Fibroblast cultures were stimulated with vehicle or lipopolysaccharide (LPS), washed, and conditioned media was collected. Fibroblast media conditioned with vehicle or LPS was applied to retrogradely labeled rat dural trigeminal ganglion neurons in vitro. Patch-clamp electrophysiology was performed to determine whether conditioned media activated/sensitized dural afferents. A preclinical behavioral model was used where conditioned media was applied directly to the rat dura to determine the presence of cutaneous facial and hind-paw allodynia. Conditioned media was also tested for interleukin-6 (IL-6) content using an enzyme-linked immunosorbent assay. Application of LPS-conditioned fibroblast media to dural afferents produced a significant increase in action potential firing as well as cutaneous facial and hind-paw allodynia when this media was applied to the dura. Finally, stimulation of cultured fibroblasts with LPS increased IL-6 levels in the media. These findings demonstrate that fibroblasts stimulated with LPS release factors capable of activating/sensitizing dural afferents. Further, they suggest that fibroblasts play a potential role in the pathophysiology of headache.     

The role of the mitochondrial protein VDAC1 in inflammatory bowel disease: a potential therapeutic target

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