Poly(ortho ester) nanoparticles targeted for chronic intraocular diseases: ocular safety and localization after intravitreal injection

Treatment of posterior eye diseases is more challenging than the anterior segment ailments due to a series of anatomical barriers and physiological constraints confronted by drug delivery to the back of the eye. In recent years, concerted efforts in drug delivery have been made to prolong the residence time of drugs injected in the vitreous humor of the eye. Our previous studies demonstrated that poly(ortho ester) (POE) nanoparticles were biodegradable/biocompatible and were capable of long-term sustained release. The objective of the present study was to investigate the safety and localization of POE nanoparticles in New Zealand white rabbits and C57BL/6 mice after intravitreal administration for the treatment of chronic posterior ocular diseases. Two concentration levels of POE nanoparticles solution were chosen for intravitreal injection: 1.5?mg/ml and 10?mg/ml. Our results demonstrate that POE nanoparticles were distributed throughout the vitreous cavity by optical coherence tomography (OCT) examination 14 days post-intravitreal injection. Intraocular pressure was not changed from baseline. Inflammatory or adverse effects were undetectable by slit lamp biomicroscopy. Furthermore, we demonstrate that POE nanoparticles have negligible toxicity assessed at the cellular level evidenced by a lack of glia activation or apoptosis estimation after intravitreal injection. Collectively, POE nanoparticles are a novel and nontoxic as an ocular drug delivery system for the treatment of posterior ocular diseases.     

Factors associated with clinically significant hypoglycemia in patients with type 1 diabetes using sensor-augmented pump therapy with predictive low-glucose management: A multicentric study on iberoamerica

Background and aimsDespite using sensor-augmented pump therapy (SAPT) with predictive low-glucose management (PLGM), hypoglycemia is still an issue in patients with type 1 Diabetes (T1D). Our aim was to determine factors associated with clinically significant hypoglycemia (<54 mg/dl) in persons with T1D treated with PLGM-SAPT.Methodology: This is a multicentric prospective real-life study performed in Colombia, Chile and Spain. Patients with T1D treated with PLGM-SAPT, using sensor ≥70% of time, were included. Data regarding pump and sensor use patterns and carbohydrate intake from 28 consecutive days were collected. A bivariate and multivariate Poisson regression analysis was carried out, to evaluate the association between the number of events of <54 mg/dl with the clinical variables and patterns of sensor and pump use.Results188 subjects were included (41 ± 13.8 years-old, 23 ± 12 years disease duration, A1c 7.2% ± 0.9). The median of events <54 mg/dl was four events/patient/month (IQR 1–10), 77% of these events occurred during day time. Multivariate analysis showed that the number of events of hypoglycemia were higher in patients with previous severe hypoglycemia (IRR1.38; 95% CI 1.19–1.61; p < 0.001), high glycemic variability defined as Coefficient of Variation (CV%) > 36% (IRR 2.09; 95%CI 1.79–2.45; p < 0.001) and hypoglycemia unawareness. A protector effect was identified for adequate sensor calibration (IRR 0.77; 95%CI 0.66–0.90; p:0.001), and the use of bolus wizard >60% (IRR 0.74; 95%CI 0.58–0.95; p:0.017).ConclusionIn spite of using advanced SAPT, clinically significant hypoglycemia is still a non-negligible risk. Only the identification and intervention of modifiable factors could help to prevent and reduce hypoglycemia in clinical practice.     

69份苦玄参种质SSR遗传多样性及品质性状相关标记分析

目的:采用简单重复序列(SSR)分子标记技术对广西苦玄参主产区69份苦玄参种质样本进行遗传多样性及亲缘关系分析,并筛选与苦玄参苷含量相关联的优良种质基因。为苦玄参种质资源评价、遗传进化分析及分子标记辅助育种等提供依据。方法:基于转录组测序技术,开发20对引物进行批量扩增。利用各标记位点的遗传多态信息,分析69份苦玄参种质的遗传多样性及亲缘关系,并通过一元线性和多元逐步回归分析,筛选与苦玄参苷含量相关联的分子标记。结果:20对SSR引物共扩增出76个等位基因,平均每个位点观测等位基因3.8个,高于有效等位基因数(1.9692个),稀有等位基因率为38.2%,等位基因分布不均匀。等位基因多态率范围为0~59%,平均38.24%,各位点多态率差异较大。各位点多态信息含量(PIC)变化范围为0~0.6211,平均0.3780;Shannon多态性信息指数变化范围为0~1.2401,平均0.7590;Nei’s基因多样性指数(Nei)变化范围为0~0.6823,平均0.4409;以上3个指标最高的为P21,最低为P7,各位点遗传多样性存在较大差异。各位点平均观测杂合度为0.3824,低于平均期望杂合度(0.4425),表现为杂合子缺失;平均遗传分化系数Fst为0.3659;基因流Nm平均值为0.4332,种质遗传分化较大,基因流较小。一元线性回归分析和多元逐步回归分析结果表明,与苦玄参苷IA和IB相关的位点各有5个,其中仅有1个位点与2个成分的含量均相关。结论:20个SSR标记位点遗传多样性存在较大的差异,供试69份种质遗传分化大,基因流较小;从供试20个SSR标记中筛选出9个与苦玄参苷含量相关联的标记位点,试验结果可为苦玄参遗传进化分析及良种选育和繁育等提供依据。     

基于1H-NMR植物代谢组学技术分析青海产区枸杞子的化学特征

目的:采用核磁共振氢谱(~1H-NMR)植物代谢组学技术比较青海产区枸杞子与其他产区(宁夏、甘肃、新疆、内蒙古)枸杞子的化学成分差异。方法:收集5个产区共97份枸杞子样本,其中青海61个样本,采用50%甲醇提取,检测~1H-NMR图谱,结合多元统计分析,对比青海产区枸杞子与其他产区枸杞子的化学差异性,并对各产区样本的枸杞多糖进行含量测定(以无水葡萄糖计),检测波长490 nm。结果:枸杞子的~1H-NMR图谱共检测到32个化学成分,多元统计分析表明青海产区枸杞子与其他产区样本相比,无明显分离趋势;青海产区枸杞子与宁夏产区相比,以及青海省6个不同地区的枸杞子相比,重叠样品较多,均不能显著分开。相似度结果表明,大多数样品的相似度0.85;化合物的单变量分析结果显示,除了蔗糖、葡萄糖、脯氨酸等个别代谢物在各产区样本中存在显著差异外,其余代谢物在各产区样品中的含量分布基本一致。青海与其他产区样本中枸杞多糖含量无显著性差异,且枸杞多糖含量与~1H-NMR指认的小分子化合物的相关系数处于-0.2～0.4。结论:采用~1HNMR植物代谢组学技术从整体化学组成上分析了青海产区枸杞子的化学特征,并结合枸杞多糖含量测定,显示青海产区枸杞子与其他产区枸杞子的化学差异较小。建立的基于~1H-NMR的枸杞子质量评价方法可为其质控水平提升及种植产区选择提供科学依据。     

基于化学模式识别技术提升参威骨痹片的质量标准并监测其生产中质控指标的转移率变化

目的:提升参威骨痹片的质量标准,初步探索其质量控制指标成分在批间含量差异较大的原因。方法:采用HPLC建立参威骨痹片的指纹图谱,以Diamonsil C18(4. 6 mm×250 mm,5μm)为色谱柱,流动相乙腈(A)-0. 1%磷酸水溶液(B)梯度洗脱(0～5 min,10%A;5～15 min,10%～12%A;15～30 min,12%～26%A;30～43 min,26%～31%A,43～50 min,31%～40%A,50～70 min,40%～55%A;70～84 min,55%～72. 5%A),检测波长230 nm。以共有峰为自变量绘制正交偏最小二乘法-判别分析-变量重要性投影(OPLS-DA-VIP)图,将共有峰对该制剂各批次间指纹图谱差异的贡献度量化,寻找差异较大的色谱峰,结合相关文献,筛选出与参威骨痹片临床适应症相关的成分并进行其含量测定的专属性试验,最终选定质控指标。通过HPLC-二极管阵列检测器(DAD)同时对本品及其生产过程中间体中质控指标进行测定,检测波长236,276,230,322 nm,其他条件同HPLC指纹图谱检测方法。结果:HPLC指纹图谱共标定了26个共有峰,各批次样品指纹图谱与对照指纹图谱的相似度均≥0. 950。优选出马钱苷酸、龙胆苦苷、芍药苷、蛇床子素为参威骨痹片的质控指标,四者的平均质量分数分别为161. 02,401. 80,255. 54,80. 68μg·g-1。结论:所建立的指纹图谱及多指标定量分析方法稳定、可靠,可用于参威骨痹片的质量控制。原料药批间质控指标成分含量差异和生产过程中间体的质控方法不够完善是引起该制剂批间质控指标成分含量差异较大的主要原因。     

针刺京骨穴联合推拿治疗背肌筋膜炎临床研究

目的探讨针刺京骨穴联合推拿治疗背肌筋膜炎的临床疗效。方法将2016年10月—2018年10月收治的背肌筋膜炎患者86例纳入研究,采用随机数字表法分组。对照组43例予以推拿治疗,观察组43例予以针刺京骨穴联合推拿治疗。比较2组患者的治疗总有效率、VAS评分及Oswestry功能障碍指数、痊愈所用时间。结果观察组治疗总有效率为95.3%,而对照组仅为81.4%,差异有统计学意义(P<0.05);2组治疗后VAS评分、Oswestry功能障碍指数均有下降,观察组上述指标低于对照组,差异有统计学意义(P<0.05);观察组痊愈所用时间短于对照组,差异有统计学意义(P<0.05)。结论针刺京骨穴联合推拿治疗背肌筋膜炎的临床疗效突出,可缓解背部疼痛、恢复功能活动,并缩短了愈合时间,提高了生活质量,值得一定的临床推广。