组成图示
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传感器类型
综述或非传感器论文
检测对象
红细胞形态异常（leptocytes、knizocytes、codocytes等）；非蛋白结合铁（NPBI）；酯化F2-异前列腺素（F2-IsoPs）；4-HNE蛋白加合物（4-HNE PAs）；动脉血氧分压（PaO2）/脉搏血氧（SpO2）；样品基质：外周血（红细胞悬液、血浆、动脉血）
检测原理
在RTT中，慢性低氧与氧化应激使血红蛋白自氧化并释放非蛋白结合铁（NPBI），引发红细胞膜磷脂中花生四烯酸自由基过氧化，形成酯化F2-异前列腺素（F2-IsoPs）；4-羟基壬烯醛（4-HNE）与膜蛋白共价结合形成4-HNE蛋白加合物（4-HNE PAs）。膜骨架与脂质损伤导致红细胞由双凹盘状变为leptocytes、knizocytes等异常形态。检测时，SEM计数形态比例；DFO螯合NPBI后经HPLC定量；GC/NICI-MS/MS检测F2-IsoPs；Western blot/ECL检测4-HNE PAs；血气与脉搏血氧评估PaO2、SpO2。氧化损伤或低氧越重，异常形态比例和标志物水平越高；ω-3 PUFAs可降低NPBI、F2-IsoPs和4-HNE PAs并部分恢复形态。
检测灵敏度
LOD: 0.1 nmol/ml；R^2 = 0.994009
效应效果
NPBI校准曲线R^2=0.994009，LOD 0.1 nmol/ml，观察者内/间变异≤2.5%/5%。RTT组红细胞内NPBI 1.6±1.0、血浆0.9±0.3 nmol/ml，高于对照0.5±0.2、0.3±0.2；F2-IsoPs（P=0.0001）和4-HNE PAs（P<0.0001）升高。leptocytes占51.83±3.5%，与IE-NPBI（r=0.6699，P=0.0174）相关。ω-3 PUFAs 6/12个月后leptocytes下降（P=0.042/0.0046），stomatocytes、discocytes增加；12个月NPBI降至0.55±0.10/0.46±0.08 nmol/ml，F2-IsoPs降50%（P=0.0002），4-HNE PAs降23.2%，PaO2正常，临床严重度降25.5%/30.1%。
传感器的构成
- 非传感器论文：未报道传感器基底或换能器电极材料，本文仅使用临床检测体系
- 样品基质：肝素抗凝血、红细胞悬液与血浆，提供红细胞形态与氧化标志物检测对象
- 形态观察层：多聚赖氨酸载玻片、Karnovsky固定液、金溅射，用于扫描电镜观察红细胞形态
- 氧化标志物检测层：去铁胺(DFO)-铁复合物、酯化F2-异前列腺素(F2-IsoPs)、4-HNE蛋白加合物(4-HNE PAs)，反映氧化损伤
- 气体交换监测层：脉搏血氧仪、血气分析仪、气体分析仪，评估低氧与肺换气
中文摘要
背景：低氧血症和氧化应激（OS）见于雷特综合征（RTT），但RTT红细胞形态未知。本研究评估RTT患者与健康受试者红细胞形态，并关联OS标志物、血氧、肺气体交换和心肺参数。方法：扫描电镜观察红细胞形态；测定红细胞内和血浆非蛋白结合铁（NPBI）、酯化F2-异前列腺素（F2-IsoPs）和4-HNE蛋白加合物（4-HNE PAs）；气体分析仪评估肺氧梯度和PaO2，脉搏血氧仪评估心肺变量；在ω-3多不饱和脂肪酸（ω-3 PUFAs）给药前后比较。结果：RTT出现leptocytes等异常形态，NPBI、膜酯化F2-IsoPs和4-HNE PAs升高；异常形态与OS标志物、肺气体交换、PaO2及心肺变量相关。ω-3 PUFAs后leptocytes减少，可逆形态增加，并伴OS损伤标志物下降、肺氧交换和心肺生理改善。结论：RTT红细胞形态改变，OS-低氧二元关系导致形态和膜损伤，ω-3 PUFAs可部分挽救。
英文摘要
BACKGROUND: Hypoxemia and increased oxidative stress (OS) have been reported in Rett Syndrome (RTT), a genetical neurodevelopmental disorder. Although OS and hypoxemia can lead to red blood cells (RBCs) shape abnormalities, no information on RBCs morphology in RTT exists. Here, RBCs shape was evaluated in RTT patients and healthy subjects as a function of OS markers, blood oxygenation, pulmonary gas exchange, and cardio-respiratory parameters.
METHODS: RBCs morphology was evaluated by Scanning Electron Microscopy. Intraerythrocyte and plasma non protein-bound iron (NPBI), esterified F(2)-Isoprostanes (F(2)-IsoPs), 4-HNE protein adducts (4-HNE PAs) were measured. Pulmonary oxygen gradients and PaO(2) were evaluated by gas analyzers and cardiopulmonary variables by pulse oximetry. In RTT patients these parameters were assessed before and after ω-3 polyunsaturated fatty acids (ω-3 PUFAs) administration.
RESULTS: Altered RBCs shapes (leptocytes) and increased NPBI were present in RTT, together with increased erythrocyte membrane esterified F(2)-IsoPs and 4-HNE PAs. Abnormal erythrocyte shapes were related to OS markers levels, pulmonary gas exchange, PaO(2) and cardio-respiratory variables. After ω-3 PUFAs, a decrease of leptocytes was accompanied by a progressive increase in reversible forms of RBCs. This partial RBCs morphology rescue was related to decreased OS damage markers, improved pulmonary oxygen exchange, and cardiopulmonary physiology.
CONCLUSIONS: These findings indicate that in RTT 1) RBCs shape is altered; 2) the OS-hypoxia diad is critical in generating altered RBCs shape and membrane damage; 3) ω-3 PUFAs are able to partially rescue RBCs morphology and the OS-derived damage.
GENERAL SIGNIFICANCE: RBCs morphology is an important biosensor for OS imbalance and chronic hypoxemia.