Highly Fluorescent Magnetic Nanobeads with a Remarkable Stokes Shift as Labels for Enhanced Detection in Immunoassays

Fluorescence immunoassays are popular for achieving high sensitivity, but they display limitations in biological samples due to strong absorption of light, background fluorescence from matrix components, or light scattering by the biomacromolecules. A powerful strategy to overcome these problems is introduced here by using fluorescent magnetic nanobeads doped with two boron‐dipyrromethane dyes displaying intense emission in the visible and near‐infrared regions, respectively. Careful matching of the emission and absorption features of the dopants leads to a virtual Stokes shift larger than 150 nm achieved by an intraparticle Förster resonance energy transfer (FRET) process between the donor and the acceptor dyes. Additionally, the magnetic properties of the fluorescent beads allow preconcentration of the sample. To illustrate the usefulness of this approach to increase the sensitivity of fluorescence immunoassays, the novel nanoparticles are employed as labels for quantification of the widely used Tacrolimus (FK506) immunosuppressive drug. The FRET‐based competitive inhibition immunoassay yields a limit of detection (LOD) of 0.08 ng mL^?1, with a dynamic range (DR) of 0.15–2.0 ng mL^?1, compared to a LOD of 2.7 ng mL^?1 and a DR between 4.1 and 130 ng mL^?1 for the immunoassay carried out with direct excitation of the acceptor dye.     

他克莫司与环磷酰胺联合激素治疗特发性膜性肾病的疗效比较

目的: 比较他克莫司( Tacrolimus, TL)与环磷酰胺( Cyclophosphamide, CTX)联合激素治疗特发性膜性肾病(idiopathic membranous nephropathy, IMN)的疗效及安全性。 方法: 选择本院原发性肾病综合征经肾活检确诊为IMN的30例患者, 排除继发性膜性肾病,随机分为两组,分别给予TL联合激素治疗[TL组,0.07～0.1 mg·kg^-1·d^-1,n=15]或CTX联合激素治疗[CTX组,0.75~1.0 g/m²,1次/月,n=15],疗程为6个月。主要观察治疗前后24小时尿蛋白、血清白蛋白、血脂、血糖、肝肾功的变化和不良反应发生率,以及治疗后的完全缓解率、部分缓解率。 结果: TL或CTX治疗6个月后疗效指标均出现明显的改善。TL组1月后24小时蛋白尿、血清白蛋白和血脂出现明显缓解,而CTX组2月后才出现明显缓解。治疗6个月后TL组完全缓解率高于CTX组(40.0% vs 13.3%),总缓解率高于CTX组(93.3% vs 60.0%)。TL组副作用主要有:胃肠不适2例(13.3%),血糖升高1例 (6.7%)、细菌性肺炎1例 (6.7%) 、带状疱疹1例 ( 6.7% ) ,血清肌酐升高1例 ( 6.7% )、 轻度肝转氨酶升高1例 (6.7% )、脱发2例(13.3%)。CTX组的副作用:胃肠道不适2例(13.3%),肺部感染2例(13.3%),带状疱疹1例(6.7%),骨髓抑制2例(13.3%),肝功能损害3例(20.0%),化学性膀胱炎2例(13.3%),脱发6例(40.0%)。 结论: TL联合激素治疗膜性肾病是有效和安全的。与CTX相比,TL能迅速缓解膜性肾病患者蛋白尿,并有更高的IMN缓解率。     

强的松联合普乐可复治疗V型狼疮性肾炎临床观察

目的对V型狼疮性肾炎的强的松联合普乐可复综合治疗进行临床分析。方法选取132例V型狼疮性肾炎患者,随机分配到对照组和实验组。每组各66例。实验组采用强的松与普乐可复的综合治疗,对照组只采用普乐可复的治疗方式。利用SPSS13.0对2组的疗效进行统计学处理。结果实验组治疗效果要明显好于对照组患者,而并发症却少于对照组患者。结论对V型狼疮性肾炎患者采用强的松联合普乐可复的综合治疗要比只单独采用普乐可复治疗更加有效,更加安全。所以研究V型狼疮性肾炎的有效治疗方式是极具临床价值的。     

Effect of aminoalkyl methacrylate copolymer E/ HCl on in vivo absorption of poorly water-soluble drug

This study aimed to investigate in vivo absorption of tacrolimus formulated as a solid dispersion using Eudragit E^®/HCl (E-SD). E-SD is an aminoalkyl methacrylate copolymer that can be dissolved under neutral pH conditions. E-SD was used alone as a solid dispersion carrier and/or was mixed with tacrolimus primarily dispersed with hydroxypropylmethylcellulose (HPMC). Tacrolimus was formulated with E-SD at several different ratios. Formulations with tacrolimus/E-SD ratio of 1/3 showed higher in vivo absorption, compared to tacrolimus dispersed in the excipients (primarily HPMC) found in commercially available tacrolimus capsules, using a rat in situ closed loop method. Good correlation was observed between in vitro drug solubility and in vivo drug absorption. In vitro solubility tests and rat oral absorption studies of tacrolimus/HPMC solid dispersion formulations were also conducted after mixing the HPMC dispersion with several ratios of E-SD. E-SD/tacrolimus/HPMC formulations yielded high in vitro drug solubility but comparatively low in vivo absorption. Dog oral absorption studies were conducted using capsules containing a formulation of tacrolimus/E-SD at a ratio of 1/5. The E-SD formulation-containing capsule showed higher in vivo drug absorption than tacrolimus dispersed in the standard HPMC capsule. These studies report enhancement of the in vivo absorption of a poorly water-soluble drug following dispersion with E-SD when compared to formulation in HPMC.     

移植后高血压与他克莫司剂量及血药浓度的关系

目的: 调查肾移植患者服用他克莫司后高血压的发生率,并探讨移植后高血压与他克莫司的服用剂量、血药浓度及血药浓度/剂量的相关性。方法: 选取以他克莫司进行治疗的肾移植患者200例,测量患者的血压。然后从中随机抽取53例高血压患者和53例正常血压患者,待患者服用他克莫司至少3 d后,运用微粒子酶免疫分析(MEIA) 法测定他克莫司谷浓度,分析他克莫司剂量、血药浓度与血压水平的相关性,并比较两组患者的他克莫司剂量、血药浓度及血药浓度/剂量的差异。结果: 在200例肾移植患者中,104例患者(52.0%)患有移植后高血压;他克莫司日剂量与患者的SBP呈正相关(r=0.216,P＜0.05),而剂量与DBP,血药浓度与血压水平均无相关性;高血压组他克莫司的服用剂量明显高于正常血压组[(3.11±1.49) mg/d∶(2.42±1.07) mg/d,P＜0.05];谷浓度两组间比较差异无统计学意义(P＞0.05);高血压组患者的谷浓度/剂量明显低于正常血压组[(2.94±1.57) ng·d/mg·mL∶(3.95±3.02) ng·d/mg·mL,P＜0.05]。结论: 肾移植患者的SBP与他克莫司日剂量具有明显相关性,服用更高剂量他克莫司的患者更易发生高血压。     

支架连接代替血管吻合建立大鼠肾移植模型

目的探讨抗淋巴细胞血清(ALS)联合他克莫司(FK506)在诱导大鼠肾脏移植免疫耐受中的作用。方法以SD大鼠作为供体,Wistar大鼠作为受体,建立大鼠肾移植模型。对照组(A组)仅行肾移植,术前术后未予免疫干预;ALS组(B组)术后当天腹腔注射ALS,连续应用至术后第10天;FK506组(C组)术后当天开始应用FK506,连续应用至术后第10天;联合组(D组)联合应用ALS与FK506。术后观测各组受者存活时间、移植肾功能、移植肾血供、免疫耐受状态等。结果D组平均存活时间为(37.0±5.3)d,与A组(7.4±1.6)d、B组(16.3±4.7)d及C组(17.5±5.3)d比较,差异均有统计学意义(P<0.05);手术后1周检查A组移植肾血供差,平均肾血管阻力指数0.80±0.06;B组和C组移植肾血供良好,平均阻力指数分别为0.62±0.07、0.63±0.08;D组移植肾血供良好,术后1周与术后20d平均阻力指数分别为0.61±0.04、0.62±0.03;D组与A组相比较差异有统计学意义(P<0.05)。结论ALS联合FK506能够延长受者存活时间和促进免疫耐受的诱导。     

窄谱中波紫外线联合他克莫司软膏治疗白癜风的临床疗效观察

目的:探讨窄谱中波紫外线联合他克莫司软膏对白癜风的治疗作用。方法:收集我院2011年至2013年门诊共90例白癜风患者,随机分为两组,每组各45例患者,分为窄谱中波紫外线联合他克莫司软膏治疗组及他克莫司软膏治疗组。随访6个月,分析两组患者治愈率,并对不同部位、不同期、不同类型白癜风的治疗情况进行比较分析。结果:窄谱中波紫外线联合他克莫司软膏对白癜风的治愈率明显高于单用他克莫司软膏组,头面部白癜风治疗有效率优于肢端部、躯干四肢部,肢端部白癜风治疗效果最差。寻常型白癜风的疗效较节段型白癜风疗效好。进展期白癜风较稳定期疗效好。结论:窄谱中波紫外线联合他克莫司软膏对白癜风具有较好的治疗作用。     

他克莫司微生物合成的研究进展

介绍了他克莫司的理化特征和免疫抑制活性,重点阐述了他克莫司发酵法生产的研究现状,着重介绍了他克莫司的合成基因簇及代谢途径研究进展,对他克莫司代谢工程改造进行综述并对今后的研究趋势进行了展望--结合系统生物学为已有的他克莫司菌株进一步进行代谢工程改造提供指导;通过合成生物学构建他克莫司合成途径全新高效的前体关键酶。     

肝移植患者他克莫司个体化给药研究

目的: 建立人工神经网络用于估算他克莫司血药浓度。 方法: 收集26例肝移植患者口服他克莫司的94份全血浓度数据,采用遗传算法配合动量法优化网络参数,建立人工神经网络。 结果: 人工神经网络平均预测误差(MPE)与平均绝对预测误差(MAE)分别为(-0.11±2.81) ng/mL 和(2.14±1.72) ng/mL,78.6%血药浓度数据绝对预测误差≤3.0 ng/mL。多元线性回归MPE与MAE分别为(0.56±2.70) ng/mL 和(2.15±1.63) ng/mL,9例次(9/14,64.3%)绝对预测误差≤3.0 ng/mL。人工神经网络准确性及精密度优于多元线性回归。 结论: 人工神经网络预测可用于预测他克莫司血药浓度,指导个体化给药。