基于裂开型核酸适体序列短、 能有效降低因探针形成二级结构产生假阳性信号等优点, 选择裂开型核酸适体作为特异性识别探针, 核酸染料噻唑橙（TO）为信号探针, 用单壁碳纳米管（SWCNTs）降低背景信号, 利用“适配体-目标分子-适配体”的“三明治”夹心方式, 建立了一种检测ATP的新方法。 在pH 80的Tris-HCl缓冲溶液中, 裂开成两段的ATP适体特异性识别ATP分子, 生成稳定的“适配体-ATP-适配体”复合结构。 单壁碳纳米管对该复合结构的吸附力较弱, 因此该复合物游离在溶液中, TO与其结合而产生强荧光。 当不存在ATP时, 核酸适体探针以单链状态存在, 可通过π—π共轭作用结合到SWCNTs表面, 进而不能与TO结合, TO游离在溶液中荧光非常微弱。 反应体系中ATP浓度越高, 形成的“适配体-ATP-适配体”夹心识别结构复合物越多, 检测到的荧光强度越大, 据此实现对ATP的检测。 在优化实验条件下, 在最大荧光发射波长550 nm处, ATP的浓度在90×10-9～10×10-7 mol·L-1范围内与ΔF/F0值成线性关系, r=0996 4。 该方法加标回收率为952%～104%, 相对标准偏差（RSD）为102%～454%, 检出限达到267×10-9 mol·L-1。 该方法基于功能核酸对目标物亲合力强、 选择识别性高的特点, 对ATP的检测表现出很好的选择性, 实验结果表明, 当相对误差控制在±5%以内时, 200倍的UTP, GTP和CTP均不干扰ATP的测定。 另外, 该方法操作简单、 快速、 无需标记、 灵敏准确, 可用于血清样品中ATP的测定, 在快速检测小分子物质领域中有较好的应用前景。

荧光测定法在化学分析和生物检测中已广泛应用, 将该法引用到中药的生物效价检测中则是一种新的尝试。 而生物效价检测与药效直接相关联, 是研究中药质量控制与品质评价方法的新趋势。 为了建立与抗病毒药效相关的板蓝根质量控制和评价方法,采用化学荧光测定法考察了板蓝根提取物对流感病毒神经氨酸酶(NA)的体外抑制活性, 分析了其药理作用的量效曲线变化趋势和作用规律。 在此基础上并根据生物检定的统计学原理, 以“质反应平行线”法设计和优化检测条件, 建立了板蓝根抗病毒生物效价荧光检测方法。 实际测试结果能够灵敏、 定量表征样品间的品质差异, 方法的可靠性和可重复性较好。 以此作为板蓝根质量生物控制和评价方法, 能够体现中药药效特点。

Severe body stress induced by hypoxemia and hypotension may lead to total body energy state deterioration. The perfusion of the most vital organs is maintained at the expense of “less vital” organs. In the present study, we used a multi-site multiparametric (MSMP) monitoring system for real-time evaluation of tissue blood flow (TBF) and mitochondrial NADH fluorescence of the brain and the small intestine following hemorrhage. In Group 1, uncontrolled hemorrhage, mean arterial pressure (MAP) was decreased to 40mmHg within 2 minutes and shed blood was re-infused after 30minutes. In Group 2, controlled hemorrhage, during the 30minutes of hemorrhage, MAP was kept at 40mmHg. During hemorrhage, in both groups, the intestinal TBF and NADH deteriorated, while the brain remained relatively well protected. In Group 1, all parameters partly recovered within the hemorrhage phase, while in Group 2, complete recovery occurred only after resuscitation. At the end of the experiment, both models showed a decrease in intestinal viability (TBF decreased, NADH increased), while the brain metabolic state in Group 2 declined slightly. Our unique multi-parametric monitoring device demonstrated that, under hemorrhage, the small intestine responded entirely differently from the brain. This may suggest the potential usefulness of the monitoring of less vital organs, as proxy organs, in critical conditions such as massive hemorrhage. The present study also highlights the importance of mitochondrial function monitoring in similar conditions in the clinical environment.