纳米材料用于放疗防护的研究进展

放射治疗是利用高能射线抑制癌细胞增殖的治疗方法,已广泛用于恶性肿瘤的治疗.但是,高能射线不可避免地会对机体的正常组织造成损害,产生放疗相关副作用.尽管目前有一些小分子放疗防护药物已应用于临床或处于临床前研究,但其较短的血液循环时间和较快的新陈代谢速度极大地削弱了其防护效果.近20年来,随着纳米技术在生物医学领域的飞速发展,纳米放疗防护剂的出现为提高防护效果提供了新的选择.通过合理地设计和开发纳米放疗防护剂,有望解决现有小分子放疗防护药物的缺陷.鉴于纳米放疗防护剂具有诸多优势,本综述概述了纳米放疗防护材料的常见设计策略,同时分析了放射诱导的常见疾病的致病机制和纳米放疗防护材料防治各种放射诱导疾病的研究现状.最后,还讨论了纳米材料用于放疗防护所面临的挑战和未来前景.     

Determination of micro‐RNA in cardiomyoblast cells using CE with LIF detection

Micro‐RNAs (miRNAs) are small, endogenous, singlestranded, and noncoding RNAs. The miRNAs have been found to perform important functions in many cellular processes, such as development, proliferation, differentiation, and apoptosis. Circulating miRNAs have been proposed as emerging biomarkers in diseases such as cancer, diabetes, and cardiovascular disease including acute myocardial infarction (AMI). In this study, we developed CE with LIF (CE‐LIF) using fluorescence‐labeled DNA probe for determination of low abundance miRNA in cell extracts. The target miRNA is miRNA‐499, a biomarker candidate of AMI with low abundance in biological samples. In order to measure the trace level of miRNA, we optimized the hybridization conditions such as hybridization time, temperature, and buffer solution. The highest fluorescence intensity of the hybridized miRNA‐499 was found when hybridization was conducted at 40°C in 50 mM Tris‐acetate (pH 8.0) buffer containing 50 mM NaCl, and 10 mM EDTA for 15 min. The hybridized miRNA‐499 was detected in cultured H9c2 cardiomyoblast cells and the analysis of miRNA‐499 was completed within 1 h using CE‐LIF. These results showed the potential of CE for fast, specific, and sensitive high‐throughput analysis of low‐abundance miRNAs in cell extracts, biofluids, and tissues.     

Urinary d‐lactate levels reflect renal function in aristolochic acid‐induced nephropathy in mice

Urinary d ‐lactate is highly correlated to diabetic nephropathy – a progressive kidney disease in renal glomeruli. In this study, we used a C3H/3e mouse model to investigate the relationship between urinary d ‐lactate and aristolochic acid nephropathy where the glomerular structure is not affected. The nephropathy was induced using intravenous injections of aristolochic acid at a dosage of 10 mg/kg per day for 5 days and was characterized biochemically and histologically. The urinary excretions of proteins, N‐acetyl‐β‐d ‐glucosaminidase and serum creatinine were determined and connected to histological conventional findings. Urinary d ‐lactate was analyzed using column‐switching high‐performance liquid chromatography with fluorescence detection. The results showed a remarkable increase of urinary markers, including of urinary proteins and N‐acetyl‐β‐d ‐glucosaminidase, and the histological examination confirmed a diagnosis of acute tubule necrosis. The ratio of d ‐lactate to creatinine in the urine of aristolochic acid‐treated mice was approximately 36 times greater than that of the mice in the control group (p < 0.05). The ratios for the two groups of mice were 311.00 ± 71.70 and 8.60 ± 1.80 µmol/mmol creatinine, respectively. These data confirm in vivo that urinary d ‐lactate reflects renal injury conditions in aristolochic acid‐treated mice and may be a marker for the assessment of nephropathy. Copyright © 2013 John Wiley & Sons, Ltd.     

Combined Delivery of a Lipopolysaccharide‐Binding Peptide and the Heme Oxygenase‐1 Gene Using Deoxycholic Acid‐Conjugated Polyethylenimine for the Treatment of Acute Lung Injury

A ternary complex comprising plasmid DNA, lipopolysaccharide‐binding peptide (LBP), and deoxycholic acid‐conjugated polyethylenimine (PEI‐DA) is prepared for combinational therapy of acute lung injury (ALI). The LBP is designed as an anti‐inflammatory peptide based on the lipopolysaccharide (LPS)‐binding domain of HMGB‐1. In vitro cytokine assays show that LBP reduces levels of proinflammatory cytokines by inhibiting LPS. PEI‐DA is synthesized as the gene carrier by conjugation of deoxycholic acid to low‐molecular weight polyethylenimine (2 kDa, PEI2k). PEI‐DA has higher transfection efficiency than high‐molecular weight polyethylenimine (25 kDa, PEI25k). The ternary complex of an HO‐1 plasmid (pHO‐1), PEI‐DA, and LBP is prepared as a combinational system to deliver the therapeutic gene and peptide. The transfection efficiency of the ternary complex is higher than that of the pHO‐1/PEI‐DA binary complex. The ternary complex also reduces TNF‐α secretion in LPS‐activated Raw264.7 macrophage cells. Administration of the ternary complex into the lungs of an animal ALI model by intratracheal injection induces HO‐1 expression and reduces levels of proinflammatory cytokines more efficiently than the pHO‐1/PEI‐DA binary complex or LBP alone. In addition, the ternary complex reduces inflammation in the lungs. Therefore, the pHO‐1/PEI‐DA/LBP ternary complex may be an effective treatment for ALI.

     

Primary study on the application of Serum Pharmacology in Chinese traditional medicine

In the paper, two main methods, which are Serum Pharmacology and Traditional Pharmacology, were adopted to study Chinese traditional medicine, such as Ginkgo biloba extract (GBE), ginsenosides (GS) and compound GG (GBE + GS), pharmacology in vitro. The results showed that there were evident difference between the results of Serum Pharmacology and that of Traditional Pharmacology. There was no significant difference between the drug effect of crude GS on nitric oxide (NO) production in ECV304 and that of crude GBE, and the drug effect of GG was superior to that of GS and GBE, respectively. But, compared with GBE serum, the GS serum up-regulation of NO production in ECV304 increased significantly, and the GG serum up-regulation of the NO production in ECV304 was inferior to that of GS serum and GBE serum significantly. The results suggested that Serum Pharmacological study should be adopted in the pharmacological investigation on the Chinese traditional medicine and the drug screening of the Chinese traditional medicine.     

Simultaneous determination of tryptophan,kynurenine, kynurenic acid,xanthurenic acid and 5‐hydroxytryptamine in human plasma by LC‐MS/MS and its application to acute myocardial infarction monitoring

Reliable methods for the determination of tryptophan and its metabolites are vital to the monitoring of biochemical states during the initiation and progression of cardiovascular disease. In the present study, a single‐run liquid chromatography–tandem mass spectrometry (LC‐MS/MS) method was developed for the simultaneous determination of tryptophan (Trp) and its metabolites, including kynurenine (Kyn), kynurenic acid (KA), xanthurenic acid (XA) and 5‐hydroxytryptamine (5‐HT), in human plasma. The plasma samples were prepared using a protein precipitation approach, and the chromatographic separation was performed by gradient elution on a C₁₈ column within a total analysis time of 3.5 min. The calibration ranges were 40–20,000 ng/mL for Trp, 4–2000 ng/mL for Kyn, 0.2–100 ng/mL for KA, 0.4–200 ng/mL for XA and 1–500 ng/mL for 5‐HT, and the precision and accuracy were acceptable. The evaluation of recovery and internal standard‐normalized matrix effect proved that the sample preparation approach was effective and the matrix effect could be negligible. The newly developed method was successfully applied to the analysis of plasma samples from healthy individuals and myocardial infarction patients. The findings suggested that the plasma concentrations of Trp, Kyn, 5‐HT as well as the concentration ratios of Kyn/Trp and Trp/5‐HT might serve as biomarkers for the monitoring of acute myocardial infarction.     

Cytochrome-c detection

Following a myocardial infarction (MI) cells die or are damaged and their contents leak into the blood circulation, resulting in elevated serum levels of various enzymes, proteins, and organic molecules. Over the past few decades, it has become standard practice to employ the detection of these elevated substances as markers for the confirmation of MIs and to monitor MI patients’ response to treatment. Although it has previously been shown that cytochrome-c, a small respiratory protein, is among those elevated, the lack of a suitable detection system has prevented its routine use in the diagnosis of MIs. We present a preliminary study in which chemiluminescence was employed to detect elevated levels of cytochrome-c in the serum of MI patients. The technique, which is specific for c-type proteins, is approx 30 times more sensitive than the traditional Coomassie blue stain and can detect as little as 0.03 μg of protein. It also has potential for diagnostic use in other diseases that are characterized by mitochondrial damage.     

丹酚酸A 对H2O2 所致大鼠脑微血管内皮细胞氧化损伤保护的实验研究

目的 观察丹酚酸A 对H2O2所致大鼠脑微血管内皮细胞（RCMECs）氧化损伤的保护作用,并探讨其可能的作用机制。方法 分离并培养大鼠脑微血管内皮细胞,用H2 O2 损伤的方法建立氧自由基损伤模型。采用丹酚酸A 进行干预后,分别测定细胞培养液中乳酸脱氢酶（LDH）活性、血栓素B2（TXB2）水平、6- 酮基前列腺素1α（6-keto-PGF1α）的含量,以及细胞内和培养液中脂质过氧化产物丙二醛（MDA）含量和超氧化物歧化酶（SOD）的活性。结果 H2O2致RCMECs 氧化损伤后,细胞LDH 释放水平、TXB2和MDA 的含量均明显增加,同时6-keto-PGF1α 含量和SOD 活性显著下降;而丹酚酸A 预处理后能呈浓度依赖性的降低RCMECs 氧化损伤后LDH 水平、TXB2含量和细胞内外的MDA 含量,提高受损细胞6-keto-PGF1α 的表达和细胞内外SOD 活性。结论 丹酚酸A 对H2O2所致RCMECs 氧化损伤具有保护作用,其机制可能与其抗氧化作用有关。     

血清CLDN5、OCLN和ZO1与颅脑损伤急性期患者损伤程度及预后的关系

目的探讨非开放性颅脑损伤急性期患者血清中封闭蛋白5（CLDN5）、密封蛋白（OCLN）和紧密连接蛋白1（ZO1）的含量与患者损伤严重程度及预后的关系。方法收集2014年2月至2015年2月非开放性颅脑损伤急性期患者92例。患者入院后立即留取外周血,检测血清中CLDN5、OCLN和ZO1的含量,分析三者与患者格拉斯哥昏迷评分（GCS）、颅内血肿体积的关系以及对患者预后的判断价值。结果不同GCS的患者间血清CLDN5和OCLN含量均有统计学差异（均P＜0.05）;不同血肿体积的患者间CLDN5和OCLN含量均有统计学差异（均P＜0.05）;死亡患者的血清CLDN5和OCLN含量明显高于存活组（均P＜0.05）,在不同GCS、血肿体积和预后的患者间比较,ZO1含量均无统计学差异（均P＞0.05）。GCS与血清CLDN5、OCLN含量均呈负相关（rCLDN5=-0.84,rOCLN=-0.85,均P＜0.01）,与ZO1无相关性（rZO1=0.14,P＞0.05）;血肿体积与血清CLDN5、OCLN和ZO1含量均呈正相关（rCLDN5=0.82,rOCLN=0.93,rZO1=0.82,均P＜0.01）。结论血清CLDN5、OCLN可以作为急性颅脑损伤患者病情严重程度和预后判断的有效生物标志物。     

Low sets without subsets of higher many‐one degree

Given a reducibility ?_r, we say that an infinite set A is r‐introimmune if A is not r‐reducible to any of its subsets B with |A\B| = ∞. We consider the many‐one reducibility ?_m and we prove the existence of a low₁ m‐introimmune set in Π⁰₁ and the existence of a low₁ bi‐m‐introimmune set.