Exogenous heat shock protein 27 uniquely blocks differentiation of monocytes to dendritic cells

Circulating heat shock protein (HSP)-27 is associated with tumor progression and increased post-injury infection. Extracellular HSP-27 might alter monocyte (MO)-derived DC and/or MPhi function to mediate immunosuppression. HSP-27 treatment inhibited expression of CD1a and CD1b/c, antigen uptake, and allogeneic T cell induction (MLR) by IL-4 + GM-CSF-differentiated human DC while increasing some MPhi characteristics ( upward arrowCD14, upward arrowCD16, upward arrowCD163). MO cytokine receptor profiles elicited by 24-h exogenous HSP-27 treatment remained supportive of immature DC (iDC) emergence ( upward arrowIL-4R, downward arrowIL-6R, downward arrowM-CSFR). IL-10, IL-6, and M-CSF (which promote MPhi differentiation) were significantly increased in IL-4 + GM-CSF + HSP-27 MO-->iDC differentiation cultures. However, HSP-27 treatment during MO differentiation to DC increased programmed cell death ligand 1 coinhibitor and depressed CD86 costimulator expression in parallel to decreased iDC MLR activity. This suggested that increased MPhi differentiation was not solely responsible for HSP-27 reduction of differentiating DC activity. HSP-27 treatment actually depressed the phagocytic capacity of MO differentiated to MPhi by IL-10 or M-CSF culture. CD163 (hemoglobin receptor) expression was depressed on M-CSF + HSP-27 MO-derived MPhi. HSP-27-mediated inhibition of MO-->iDC differentiation was reversed by p38alpha & beta inhibitor (SB202190) addition or TLR4 receptor modulation. HSP-27 impaired appropriate MO-->iDC and MO-->MPhi differentiation modulating expression of receptors necessary for their proper functions. This suggests that endogenous HSP-27 has immunoregulatory activities which could contribute to immunopathology.     

EphrinB1-EphB signaling regulates thymocyte-epithelium interactions involved in functional T cell development

The Eph and ephrin families are involved in numerous developmental processes. Recently, an increasing body of evidence has related these families with some aspects of T cell development. In the present study, we show that the addition of either EphB2-Fc or ephrinB1-Fc fusion proteins to fetal thymus organ cultures established from 17-day-old fetal mice decreases the numbers of both double-positive (CD4(+)CD8(+)) and single-positive (both CD4(+)CD8(-) and CD4(-)CD8(+)) thymocytes, in correlation with increased apoptosis. By using reaggregate thymus organ cultures formed by fetal thymic epithelial cells (TEC) and CD4(+)CD8(+) thymocytes, we have also demonstrated that ephrinB1-Fc proteins are able to disorganize the three-dimensional epithelial network that in vivo supports the T cell maturation, and to alter the thymocyte interactions. In addition, in an in vitro model, Eph/ephrinB-Fc treatment also decreases the formation of cell conjugates by CD4(+)CD8(+) thymocytes and TEC as well as the TCR-dependent signaling between both cell types. Finally, immobilized EphB2-Fc and ephrinB1-Fc modulate the anti-CD3 antibody-induced apoptosis of CD4(+)CD8(+) thymocytes in a process dependent on concentration. These results therefore support a role for Eph/ephrinB in the processes of development and selection of thymocytes as well as in the establishment of the three-dimensional organization of TEC.     

Rutaecarpine attenuates osteoclastogenesis by impairing macrophage colony stimulating factor and receptor activator of nuclear factor κ‐B ligand‐stimulated signalling pathways

Rutaecarpine is a major alkaloid isolated from Evodia rutaecarpa. Here, we investigated the effects of rutaecarpine on osteoclast differentiation induced by macrophage colony stimulating factor (M‐CSF) and receptor activator of nuclear factor κ‐B ligand (RANKL) in bone marrow‐derived macrophages (BMMs). Treatment with rutaecarpine significantly inhibited osteoclastogenesis and prevented bone resorption of BMM‐derived osteoclasts. Mechanistically, rutaecarpine decreased the protein level of nuclear factor of activated T cells cytoplasmic‐1 (NFATc1) and the phosphorylation of other signalling pathways during the osteoclast differentiation. Thus, rutaecarpine may be useful as a therapeutic agent for the treatment of bone diseases.     

琥珀酸美托洛尔缓释片在健康中国人体空腹和高脂餐后单次给药的药代动力学研究

目的评价单次口服琥珀酸美托洛尔缓释片在中国健康受试者体内的药代动力学特征和安全性。方法 7例中国健康志愿者空腹单次口服琥珀酸美托洛尔缓释片47. 5 mg,8例中国健康志愿者在高脂餐后单次口服琥珀酸美托洛尔缓释片47. 5 mg,用液相色谱-串联质谱联用法测定给药后不同时间美托洛尔的血药浓度,并用Win Nonlin 6. 4软件计算主要药代动力学参数。结果本研究未观察到药物相关的不良事件和严重不良事件。空腹单次口服琥珀酸美托洛尔缓释片后的药代动力学参数如下:C_max为(14. 63±8. 93) ng·m L^-1,t_max为(9. 43±2. 76) h,t1/2为(9. 36±5. 76) h,AUC_0-t为(343. 63±262. 07) ng·m L^-1·h,AUC_inf为(359. 65±259. 29) ng·m L^-1·h,清除率为(188. 58±109. 10) L·h^-1,表观分布容积为(3031. 36±4029. 78) L。高脂餐后单次口服琥珀酸美托洛尔缓释片后的药代动力学参数如下:C_max为(19. 56±14. 89) ng·m L^-1,t_max(6. 00±1. 07) h,t1/2为(6. 27±1. 37) h,AUC_0-t为(391. 07±307. 06) ng·m L^-1·h,AUC_inf为(395. 84±311. 42) ng·m L^-1·h,清除率为(225. 97±201. 53) L·h^-1,表观分布容积为(2022. 20±1842. 51) L。结论高脂餐后给予琥珀酸美托洛尔缓释片,t_max较空腹给药提前,暴露量未见明显差异。     

靶向下调上皮细胞黏附分子对结直肠癌干细胞功能及药物敏感性的影响

目的研究靶向下调上皮细胞黏附分子(ECAM)对结直肠癌干细胞增殖、侵袭及药物敏感性的影响。方法用肿瘤微球法从人结直肠癌细胞系LoVo中获取结直肠癌干细胞,用结直肠癌干细胞表面特殊标志物[ECAM和人类细胞分化抗原44(CD44)]对其进行鉴定并进行后续研究。用lipofectamine 2000脂质体介导完成转染,根据处理方法不同将细胞分为3组:实验组(ECAM抑制组),对照组(转染公共抑制剂lipofectamine 2000-抑制剂),空白组(不作任何处理),并进行培养,取3组对数期的细胞分别给予几个浓度(1,5,10,15,20,25,30,35,40,45,50 mg·L^-1)伊立替康和几个浓度(50,100,150,200,250,300,350,400,450,500,550,600 mg·L^-1)卡培他滨,继续培养。用实时荧光定量检测用药前后3组细胞中ECAM mRNA的表达水平,用噻唑蓝(MTT)法检测用药前后3组细胞的增殖能力和药物敏感性,用Transwell小室检测3组细胞的侵袭能力。结果在富集前后的LoVo细胞系中,EpCAM^+CD44^+双阳结直肠癌干细胞的百分率分别是0.95%,85.78%,与富集前比较差异均有统计学意义(均P<0.05)。空白组、对照组和实验组中EpCAM mRNA的表达水平分别为8.17±0.64,7.94±0.83,2.16±0.12,对照组和实验组与空白组比较,差异均有统计学意义(P<0.05,P<0.01);实验组与对照组比较,差异有统计学意义(P<0.05),说明3组细胞构建成功。空白组、对照组和实验组中侵袭细胞分别为79.22±5.25,80.12±4.89,31.23±2.36。对照组和实验组与空白组比较,差异均有统计学意义(P<0.05,P<0.01);实验组与对照组比较,差异有统计学意义(P<0.05)。空白组、对照组与实验组的伊立替康对结直肠癌干细胞的IC₅₀分别为(20.25±4.35),(19.22±3.99),(10.24±2.04)mg·L^-1;这3组的卡培他滨对结直肠癌干细胞的IC₅₀分别为(320.13±23.65),(315.79±21.03),(250.22±15.45)mg·L^-1,实验组与对照组比较,差异有统计学意义(P<0.05)。结论靶向下调ECAM可以有效地抑制结肠癌干细胞增殖及侵袭能力,同时增强其对药物的敏感性。     

Nanomagnetic Modulation of Tumor Redox State

Modulation of reactive oxygen and nitrogen species in a tumor could be exploited for nanotherapeutic benefits. We investigate the antitumor effect in Walker-256 carcinosarcoma of magnetic nanodots composed of doxorubicin-loaded Fe₃O₄ nanoparticles combined with electromagnetic fields. Treatment using the magnetic nanodot with the largest hysteresis loop area (3402 erg/g) had the greatest antitumor effect with the minimum growth factor 0.49 ± 0.02 day^–1 (compared to 0.58 ± 0.02 day^–1 for conventional doxorubicin). Electron spin resonance spectra of Walker-256 carcinosarcoma treated with the nanodots, indicate an increase of 2.7 times of free iron (that promotes the formation of highly reactive oxygen species), using the nanodot with the largest hysteresis loop area, compared to conventional doxorubicin treatment as well as increases in ubisemiquinone, lactoferrin, NO-FeS-proteins. Hence, we provide evidence that the designed magnetic nanodots can modulate the tumor redox state. We discuss the implications of these results for cancer nanotherapy.     

Medical service provider networks

In many countries, health insurers or health plans choose to contract either with any willing providers or with preferred providers. We compare these mechanisms when two medical services are imperfect substitutes in demand and are supplied by two different firms. In both cases, the reimbursement is higher when patients select the in‐network provider(s). We show that these mechanisms yield lower prices, lower providers' and insurer's profits, and lower expense than in the uniform‐reimbursement case. Whatever the degree of product differentiation, a not‐for‐profit insurer should prefer selective contracting and select a reimbursement such that the out‐of‐pocket expense is null. Although all providers join the network under any‐willing‐provider contracting in the absence of third‐party payment, an asymmetric equilibrium may exist when this billing arrangement is implemented.     

Valuation of EuroQol Five-Dimensional Questionnaire,Youth Version (EQ-5D-Y) and EuroQol Five-Dimensional Questionnaire,Three-Level Version (EQ-5D-3L) Health States: The Impact of Wording and Perspective

Background

Valuations of health states were affected by the wording of the two instruments (EQ-5D-3L and EQ-5D-Y) and by the perspective taken (child or adult).

夏永良应用“玄府郁闭”理论治疗青春期后痤疮的经验浅析

[目的] 基于“玄府郁闭”理论,探讨夏永良老师治疗湿热郁闭型青春期后痤疮的临床经验。[方法] 通过查阅《黄帝内经》《素问玄机原病式》及后世医药典籍,分析“玄府郁闭”的理论依据,收集整理夏师门诊诊治的湿热郁闭型青春期后痤疮病例,并总结相关治疗经验及用药特色,最后分析典型案例进一步论述夏师临证经验。[结果] 夏师认为,湿热郁火之邪可致玄府闭塞、气津不通而发生痤疮,治疗中应以麻黄连轺赤小豆汤为基础,清里之湿热,开表之玄府,畅其气津,祛其郁阻,则痤疮可消。所举验案,夏师辨为湿热内蕴、经络瘀滞之证,治拟清热利湿、宣郁通络之法,方以麻黄连轺赤小豆汤合升降散、葛根芩连汤加减,辄收佳效。[结论] 夏师治疗湿热郁闭型青春期后痤疮以开通玄府为基础,标本兼顾,并注重食饮有节,起居有常,其临床经验值得学习和借鉴。     

THSD4基因在小鼠间充质干细胞及MC3T3-E1细胞成骨分化中的作用

目的 探讨THSD4基因对小鼠间充质干细胞和MC3T3-E1细胞成骨分化的影响。方法 提取绝经后骨质疏松症患者的骨髓间充质干细胞进行基因测序分析,与骨关节炎患者的骨髓间充质干细胞进行比较,分析基因表达差异。通过提取不同分化阶段的小鼠骨髓间充质干细胞（M-BMSC）及MC3T3-E1细胞的mRNA来检测THSD4 基因以及成骨分化的标志性基因（ALP、Runx2、Osx）的表达水平。通过构建慢病毒表达载体来实现对M-BMSC及MC3T3-E1细胞中THSD4的敲减及过表达,并观察其对M-BMSC及MC3T3-E1细胞成骨分化能力的影响。结果 THSD4基因在绝经后骨质疏松症患者骨髓间充质干细胞中明显下调,且通过KEGG以及GO富集分析发现THSD4基因可能与PI3K-AKT信号通路及Wnt信号通路相关。随着成骨诱导分化时间的延长,THSD4 mRNA和成骨分化标志性基因（ALP、Runx2、Osx）mRNA在MC3T3-E1以及M-BMSC中表达量均逐渐增加。过表达THSD4可以增强MC3T3-E1细胞和M-BMSC的成骨分化能力,而敲减THSD4则减弱了MC3T3-E1细胞和M-BMSC的成骨分化能力。结论 THSD4基因在绝经后骨质疏松症患者骨髓间充质干细胞中明显下调,且THSD4基因可以增强MC3T3-E1细胞以及M-BMSC的成骨分化能力。