Morphine or its withdrawal affects plasma malondialdehyde,vitamin E levels and absence or presence of abstinence signs in rats

Objectives Various experimental observations show that morphine treatment generates reactive oxygen species, and that its discontinuation leads to signs of withdrawal. We therefore investigated plasma malondialdehyde and vitamin E levels under both conditions to verify the occurrence of any alterations in oxidative metabolism, and whether these are associated with behavioural changes. Methods We investigated the effects of morphine or morphine plus naloxone on plasma malondialdehyde, vitamin E levels and withdrawal signs such as jumping, wet dog shakes and faecal excretion in rats. Furthermore, isopropylnoradrenaline was injected in rabbits to verify its effects on plasma malondialdehyde levels. Key findings Morphine treatment increased free malondialdehyde and decreased vitamin E levels. The elevation in malondialdehyde levels were exacerbated by the abrupt removal of morphine by naloxone, which also led to the appearance of withdrawal signs. The increased malondialdehyde values can be attributed to the interactions of reactive oxygen species with unsaturated fatty acids, and the lowered levels of vitamin E to its interactions with reactive oxygen species. Conclusions A connection seems to exist between altered peroxide status and withdrawal signs in abstinent animals.     

Cytotoxicity,oxidative stress and inflammation induced by ZnO nanoparticles in endothelial cells: interaction with palmitate or lipopolysaccharide

Recent studies showed that ZnO nanoparticles (NPs) might induce the toxicity to human endothelial cells. However, little is known about the interaction between ZnO NPs and circulatory components, which is likely to occur when NPs enter the blood. In this study, we evaluated ZnO NP‐induced cytotoxicity, oxidative stress and inflammation in human umbilical vein endothelial cells (HUVECs), with the emphasis on the interaction with palmitate (PA) or lipopolysaccharide (LPS), because PA and LPS are normal components in human blood that increase in metabolic diseases. Overall, ZnO NPs induced cytotoxicity and intracellular reactive oxygen species (ROS) at a concentration of 32 μg ml⁻¹, but did not significantly affect the release of inflammatory cytokines or adhesion of THP‐1 monocytes to HUVECs. In addition, exposure to ZnO NPs dose‐dependently promoted intracellular Zn ions in HUVECs. PA and LPS have different effects. Two hundred μm PA significantly induced cytotoxicity and THP‐1 monocyte adhesion, but did not affect ROS or release of inflammatory cytokines. In contrast, 1 μg ml⁻¹ LPS significantly induced ROS, release of inflammatory cytokines and THP‐1 monocyte adhesion, but not cytotoxicity. The presence of ZnO NPs did not significantly affect the toxicity induced by PA or LPS. In addition, the accumulation of Zn ions after ZnO NP exposure was not significantly affected by the presence of PA or LPS. We concluded that there was no interaction between ZnO NPs and PA or LPS on toxicity to HUVECs in vitro . Copyright © 2016 John Wiley & Sons, Ltd.     

Gender-dependent dissociation between oxytocin but not ACTH, cortisol or TSH responses to m-chlorophenylpiperazine in healthy subjects

m-chlorophenylpiperazine (m-CPP), a serotonin (5-HT) agonist with some selectivity for the 5-HT_2C receptor subtype, which is widely used to examine 5-HT receptor function in human subjects, has been found to induce oxytocin and thyrotropin (TSH) responses in rodents. This study examined whether m-CPP had any effect on plasma oxytocin, TSH and aldosterone concentration in healthy volunteers using a double-blind, placebo-controlled crossover design. Plasma adrenocorticorticotropic hormone (ACTH) and cortisol responses, two generally accepted markers of m-CPP-induced 5-HT receptor activation, were measured in parallel. Male subjects (n = 7) received placebo, 0.25 and 0.5 mg/kg oral m-CPP. In female subjects (n = 5), the effects of placebo and 0.25 mg/kg m-CPP were studied. After placebo, given in the morning, ACTH, cortisol, TSH and aldosterone concentrations decreased over time. m-CPP 0.25 mg/kg avoided decreases in ACTH, cortisol and TSH concentrations; these responses were significant. At the dose of 0.5 mg/kg, m-CPP caused increase in ACTH, cortisol, TSH and aldosterone concentrations. Significant plasma oxytocin responses were found in female subjects only; thus this effect of m-CPP was statistically significantly gender dependent. Other responses to m-CPP were similar in male and female subjects. The present results suggest that there are clear differences, including dose and gender-dependent dissociations, among the 5-HT receptor agonist m-CPP-induced neuroendocrine responses. Received: 6 June 1997/Final version: 27 October 1997     

Toxicity of ZnO nanoparticles (NPs) with or without hydrophobic surface coating to THP-1 macrophages: interactions with BSA or oleate-BSA

It is recently shown that biological macromolecules in food could interact with nanoparticles (NPs) and consequently change the biological effects of NPs. In this study, the interactions between ZnO NPs with or without hydrophobic surface coating and bovine serum albumin (BSA) or oleate (OA) complexed to BSA (OA-BSA) were assessed. Atomic force microscope (AFM) showed topographic changes of both types of NPs by BSA or OA-BSA, which could indicate the formation of protein corona. ZnO NPs showed negative Zeta potential, which was slightly decreased by BSA or OA-BSA, with OA-BSA being more effective. The UV–Vis was increased, whereas the fluorescence and synchronous fluorescence was decreased by the presence of ZnO NPs. Exposure to both types of ZnO NPs was associated with cytotoxicity to THP-1 macrophages, which was equally mitigated by BSA or OA-BSA associated with decreased cellular Zn elements. Exposure to ZnO NPs was associated with decreased release of cytokines, which was not affected by BSA or OA-BSA. In combination, the results from this study suggested that both BSA and OA-BSA could be adsorbed to ZnO NPs regardless of hydrophobic surface coating, which reduced the cytotoxicity of NPs to macrophages probably due to reduced association between NPs and cells. BSA and OA-BSA equally protected THP-1 macrophages from ZnO NP exposure, which might indicate that complexation to OA did not compromise the cytoprotective effects of BSA. These data might also indicate the complex interaction between NPs and biological macromolecules as food components, which should be considered for future nanotoxicological studies.     

Genes Expressed in Cell Types after Their Exposure to UV Stresses or Oxidative Stresses,e.g., Keratinocytes,Fibroblasts in the Skin,or Endothelial Cells,Neurones or Astrocytes in Age-Related Pathologies Where Oxidative Stresses have been Shown to Occur,such as Atherosclerosis,Parkinson's Disease,Alzheimer's Disease

Introduction: The IGF system controls growth, differentiation, and development at the cellular, organ and organismal levels. IGF1 receptor (IGF1R) signaling is dysregulated in many cancers. Numerous clinical trials are currently assessing therapies that inhibit either growth factor binding or IGF1R itself. Therapeutic benefit, often in the form of stable disease, has been reported for many different cancer types.

Areas covered: Canonical IGF signaling and non-canonical pathways involved in carcinogenesis. Three recent insights into IGF1R signaling, namely hybrid receptor formation with insulin receptor (INSR), insulin receptor substrate 1 nuclear translocation, and evidence for IGF1R/INSR as dependence receptors. Different approaches to targeting IGF1R and mechanisms of acquired resistance. Possible mechanisms by which IGF1R signaling supports carcinogenesis and specific examples in different human tumors.

Expert opinion: Pre-clinical data justifies IGF1R as a target and early clinical trials have shown modest efficacy in selected tumor types. Future work will focus upon assessing the usefulness or disadvantages of simultaneously targeting the IGF1R and INSR, biomarker development to identify potentially responsive patients, and the use of IGF1R inhibitors in combination therapies or as an adjunct to conventional chemotherapy.     

Role of AHR2 in the expression of novel cytochrome P450 1 family genes, cell cycle genes, and morphological defects in developing zebra fish exposed to 3,3',4,4',5-pentachlorobiphenyl or 2,3,7,8-tetrachlorodibenzo-p-dioxin.

Halogenated agonists for the aryl hydrocarbon receptor (AHR), such as 3,3',4,4',5-pentachlorobiphenyl (PCB126) and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), cause developmental toxicity in fish. AHR dependence of these effects is known for TCDD but only presumed for PCB126, and the AHR-regulated genes involved are known only in part. We defined the role of AHR in regulation of four cytochrome P450 1 (CYP1) genes and the effect of PCB126 on cell cycle genes (i.e., PCNA and cyclin E) in zebra fish (Danio rerio) embryos. Basal and PCB126-induced expression of CYP1A, CYP1B1, CYP1C1, and CYP1C2 was examined over time as well as in relation to cell cycle gene expression and morphological effects of PCB126 in developing zebra fish. The four CYP1 genes differed in the time for maximal basal and induced expression, i.e., CYP1B1 peaked within 2 days postfertilization (dpf), the CYP1Cs around hatching (3 dpf), and CYP1A after hatching (14-21 dpf). These results indicate developmental periods when the CYP1s may play physiological roles. PCB126 (0.3-100nM) caused concentration-dependent CYP1 gene induction (EC50: 1.4-2.7nM, Lowest observed effect concentration [LOEC]: 0.3-1nM) and pericardial edema (EC50: 4.4nM, LOEC: 3nM) in 3-dpf embryos. Blockage of AHR2 translation significantly inhibited these effects of PCB126 and TCDD. PCNA gene expression was reduced by PCB126 in a concentration-dependent manner, suggesting that PCB126 could suppress cell proliferation. Our results indicate that the four CYP1 genes examined are regulated by AHR2 and that the effect of PCB126 on morphology in zebra fish embryos is AHR2 dependent. Moreover, the developmental patterns of expression and induction suggest that CYP1 enzymes could function in normal development and in developmental toxicity of PCB126 in fish embryos.     

Estimation of AUC or Partial AUC Under Test-Result-Dependent Sampling

The area under the receiver operating characteristic (ROC) curve (AUC) and partial area under the ROC curve (pAUC) are summary measures used to assess the accuracy of a biomarker in discriminating true disease status. The standard sampling approach used in biomarker validation studies is often inefficient and costly, especially when ascertaining the true disease status is costly and invasive. To improve efficiency and reduce the cost of biomarker validation studies, we consider a test-result-dependent sampling (TDS) scheme, in which subject selection for determining the disease state is dependent on the result of a biomarker assay. We first estimate the test-result distribution using data arising from the TDS design. With the estimated empirical test-result distribution, we propose consistent nonparametric estimators for AUC and pAUC and establish the asymptotic properties of the proposed estimators. Simulation studies show that the proposed estimators have good finite sample properties and that the TDS design yields more efficient AUC and pAUC estimates than a simple random sampling design. A data example based on an ongoing cancer clinical trial is provided to illustrate the TDS design and the proposed estimators. This work can find broad applications in the design and analysis of biomarker validation studies.     

Opposed arsenite-mediated regulation of p53-survivin is involved in neoplastic transformation, DNA damage, or apoptosis in human keratinocytes

Biphasic dose-response relationship induced by environmental agents is often characterized with the effect of low-dose stimulation and high dose inhibition. Some studies showed that arsenite may induce cell proliferation and apoptosis via biphasic dose-response relationship in human cells; however, mechanisms underlying this phenomenon are not well understood. Our present study shows that, for human keratinocytes (HaCaT) cells, a low concentration of arsenite activates extracellular signal-regulated kinases (ERKs), which leads to up-regulation of nuclear factor κB (NF-κB) binding to DNA and to elevated, NF-κB-dependent expression of mot-2 (a p53 inhibitor) and survivin (an inhibitor of apoptosis). Activation of p53 is blocked, and neoplastic transformation is enhanced. Inhibition of ERKs reduces cell proliferation and neoplastic transformation. In contrast, a high concentration of arsenite activates c-Jun N-terminal kinases (JNKs), positive regulators of p53, by binding to p53 and preventing its murine double minute 2 (mdm2)-mediated degradation. The elevated levels of p53 lead to repair of DNA damage and apoptosis. Inhibition of JNKs increases DNA damage but decreases apoptosis. By identifying a mechanism whereby ERKs and JNKs-mediated regulation of the p53-survivin signal pathway is involved in the biphasic effects of arsenite on human keratinocytes, our data expand understanding of arsenite-induced cell proliferation, neoplastic transformation, DNA damage, and apoptosis.     

类风湿性关节炎患者服用双氯芬酸、萘丁美酮、美洛昔康、塞来昔布6mon生存质量的队列研究

目的　对双氯芬酸、萘丁美酮、美洛昔康、塞来昔布治疗类风湿性关节炎 6mon前后患者生命质量进行评估。方法　采用回顾性和前瞻性队列研究相结合的方式 ,随机选择类风湿性关节炎新发病例 4 6 1名 ,其中双氯芬酸组 ,萘丁美酮组各 131名 ,美洛昔康组 14 4名 ,塞来昔布组 5 5例。观察时间 6mon。采用SF 36量表 (中文版 )对患者服药前后的健康相关生存质量进行评估 ,该评估包括躯体功能、躯体角色、肌体疼痛、总体健康状况、活力、社会功能、情绪角色和心理卫生 8个分量表。结果　按分量表进行分析 ,结果显示四组服药前后除“社会功能”的改变、以及塞来昔布组服药前后“躯体角色”的改变上无显著差异外 ,其他分量表各组服药前后均有显著差异。各分量表服药前后改变 4组间差异无显著性。按每个条目进行分析结果显示 ,双氯芬酸组在总体健康状况的改善 (条目 2 )、自我感觉身体疼痛改善程度 (条目7)以及SF 36量表总评分改善上优于塞来昔布组 (P均 <0 0 5 )。临床控制组生存质量改变优于其他 3组 ,显效组和好转组之间差异无显著性而且均优于无效组。结论　生存质量评分的改变与药物疗效呈正相关 ,SF 36量表 (中文版 )能用来评估非甾体抗炎药治疗前后生存质量的改变。按分量表进行分析四种NSAIDs前后患者生存质     

The pathogenesis of,and pharmacological treatment for,Canavan disease

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Effects of MPEP on locomotion, sensitization and conditioned reward induced by cocaine or morphine

Exposure to environmental cues is considered a major cause of relapse in detoxified addicts. Recent findings showed an involvement of glutamate in cue-induced relapse and suggest that subtype 5 of metabotropic glutamate receptors (mGluR5) is involved in conditioned drug-reward. The present study applied the conditioned place preference (CPP) paradigm to examine the involvement of mGluR5 in cocaine- and morphine-induced behaviours. Results of previous mice-studies were extended into rats by using the selective mGluR5 antagonist 2-methyl-6-(phenylethynyl)pyridine (MPEP). As a result, the evaluated behavioural parameters were dose-relatedly affected by MPEP. Low-dosed MPEP (10 mg/kg, i.p.) did not affect spontaneous locomotion, reduced cocaine-induced hyperlocomotion and produced sensitized locomotion, while showing no effect on sensitized locomotion induced by repeated cocaine or morphine. Low-dosed MPEP did not genuinely block development of cocaine- and morphine-CPP, but rendered CPP expression state-dependent. The medium MPEP-dose (30 mg/kg) was most effective in reducing spontaneous locomotion. The high MPEP-dose (50 mg/kg) was most effective in reducing both body-weight and morphine-CPP expression. Cocaine-CPP expression was not affected by any MPEP-dose. In conclusion, mGluR5 are involved in modulation of spontaneous and cocaine-induced locomotion, in state-dependent learning and in expression of morphine-CPP. Thus, MPEP may be beneficial for relapse prevention in morphine-addicts.     

不同药物用于悬雍垂腭咽成形术后静脉自控镇痛的比较

目的 观察曲马多和和芬太尼用于悬雍垂腭咽成形术（UPPP）后患者自控镇痛（PCA）的有效性及安全性.方法 择期拟行UPPP的OSAS患者40例,ASAⅠ～Ⅱ级,随机分为两组,每组20例.曲马多组：术后使用静脉曲马多自控镇痛 芬太尼组：术后使用静脉芬太尼自控镇痛.曲马多组PCA以20 mg/h背景剂量泵入曲马多,负荷剂量为50 mg,单次PCA剂量为20mg,锁定时间30min 芬太尼组PCA以20 μg/h背景剂量泵入芬太尼,负荷剂量为50μg,单次PCIA剂量为20 μg,锁定时间30min.比较患者术后2、4、8、16、24h的动脉血气、血糖水平、镇痛效果、恶心呕吐情况以及MBP、HR、RR、SpO2的变化.结果 两组患者术后无一例呼吸暂停,SpO2均在93%以上,血气均在正常范围内.与术前基础值比较,两组患者在术后各时间点MBP、HR、SpO2、血糖无显著性变化,镇痛效果良好,术后各时间点VAS评分显著性降低 但与曲马多组比较,芬太尼组有呼吸抑制.结论 曲马多较芬太尼用于悬雍垂腭咽成形术后静脉自控镇痛时,安全有效、切实可行.     

No changes in event-related potentials with estrogen or estrogen plus progesterone treatment in healthy older hysterectomized women: results from a double-blind,placebo-controlled study

Rationale The potential to improve cognition in older women with estrogen or estrogen/progesterone therapy is currently a matter of intense debate. Only a few studies conducted so far have used electrophysiological indicators of cognitive information processing as outcome measures in randomised placebo controlled studies.Objectives This study was undertaken to measure changes in event-related potentials (ERPs) after short (4 weeks) or prolonged (24 weeks) hormone treatment in older women.Methods A randomised, double-blind, placebo-controlled study in hysterectomized older women (aged 58–75 years) was performed (n=51). The participants received orally estradiol (2 mg estradiol valerate), estradiol plus progesterone (100 mg micronized progesterone) or placebo for 24 weeks. Using four different paradigms, early and late ERPs were assessed at baseline and after 4 and 24 weeks of treatment.Results Strong hormone increases were observed in the two active treatment groups. However, no significant effects on any of the assessed ERPs were observed in either of the two treatment groups. Similar non-significant findings were obtained for reaction time and error rate.Conclusions Estradiol or estradiol/progesterone treatment appears to have no strong effects on several ERP markers of information processing in older hysterectomized women. The current negative findings might suggest a reduced sensitivity of the aged brain to gonadal steroids.     

Fluoxetine,but not sertraline or citalopram,potentiates the locomotor stimulant effect of cocaine: possible pharmacokinetic effects

Rationale The selective serotonin reuptake inhibitor (SSRI) fluoxetine enhances some of the behavioural effects of cocaine, including locomotor stimulation. While this effect has often been interpreted as evidence for a serotonergic component to the behavioural effects of cocaine, direct evidence for this hypothesis is lacking. One alternative explanation is that fluoxetine, by inhibiting cytochrome P450 (CYP) enzymes, interferes with the metabolism of cocaine.Objectives These experiments were undertaken to: 1) compare the effects of fluoxetine with those of two other SSRIs, sertraline and citalopram, on cocaine-induced locomotor activity, 2) examine the effects of fluoxetine on cocaine-stimulated locomotion in rats depleted of serotonin (5-hydroxytrptamine; 5-HT), and 3) determine the effect of fluoxetine on cocaine levels in the brain.Methods Locomotor activity was measured, using photocell based activity monitors, in rats habituated to those monitors. Depletion of 5-HT was achieved by injecting 5,7-dihydroxytryptamine (5,7-DHT) into the dorsal and median raphe nuclei. Cocaine levels in whole brain were measured using high-performance liquid chromatography with ultraviolet detection.Results In experiment 1, 5 mg/kg fluoxetine enhanced the ability of 10 and 15 mg/kg cocaine to increase locomotor activity. Neither citalopram nor sertraline (5 and 10 mg/kg) altered the stimulant effect of 10 mg/kg cocaine. Experiment 2 showed that this effect of fluoxetine was also apparent in rats with large and widespread depletion of brain 5-HT levels. The 5-HT depletion also failed to alter the response to cocaine itself. In experiment 3, brain levels of cocaine were elevated in rats pretreated with fluoxetine compared with rats that received cocaine alone.Conclusion Fluoxetine enhanced the ability of cocaine to increase locomotor activity. This effect appears not to depend upon increasing 5-HT function since fluoxetine was also effective in rats with substantial 5-HT depletions, and two other SSRIs did not alter the effects of cocaine. Fluoxetine-treated rats had higher brain levels of cocaine than rats treated with cocaine alone. This effect suggests that fluoxetine slows the metabolism of cocaine, perhaps by inhibition of CYP enzymes involved in metabolizing cocaine. The results also indicate that 5-HT reuptake inhibition may not play a prominent role in mediating the stimulant effects of cocaine.     

Role of TLR4 and TCR or BCR against baicalin-induced responses in T and B cells

Baicalin (BA), a flavonoid compound isolated from Scutellaria baicalensis, has been shown to possess a number of pharmacological effects including antiviral, anti-inflammatory, antioxidant and immune regulation. Here, we examined its effects on human T and B cells proliferation by MTT assay and found that BA stimulated T and B cells proliferation, independently and cooperatively with Con-A (T cells) or LPS (B cells). Then, we analyzed the effects of BA treatment on the mRNA expression of Toll-like receptors (TLRs), IL-2, IFN-γ and IL-12 in T and B cells by real-time RT-PCR and attempted to observe whether blocking TLR4 had influence on mRNA expression. We found that BA treatment significantly up-regulated TLR3, 7, 8 and 9 mRNA expressions in T and B cells, IL-2 and IFN-γ in T cells and IL-12 in B cells. The increased mRNA expressions were suppressed after blocking TLR4. We further analyzed the effects of BA treatment on TCR vβ and CD79 mRNA expression levels in T and B cells and explored whether blocking TCR (αβ) or BCR mIgM F(ab′)₂ had an influence on mRNA expression. We found that BA treatment significantly improved TCR vβ and CD79 mRNA expression in T and B cells, respectively, and the improvements were all inhibited after blocking TCR (αβ) or BCR mIgM F(ab′)₂. Our results suggested that BA participates in innate and adaptive immune regulation by up-regulating the mRNA expression of TLRs (3, 7, 8 and 9), IL-2, IFN-γ and IL-12 in T and B cells, which is mediated by TLR4, and by improving the mRNA expression of TCR vβ and CD79, which is mediated by TCR (αβ) and BCR mIgM, respectively. Therefore, TLR4, TCR (αβ) and BCR mIgM are all the immune receptors for BA on T and B cells.     

硒和/或维生素E对实验性高脂血症大鼠心、肝、肾及血清中一氧化氮及一氧化氮合酶的影响

目的 :观察硒和/或维生素E(VE)对实验性高脂血症大鼠心、肝、肾、血清一氧化氮 (NO)及一氧化氮合酶 (NOS)的影响。方法 :对大鼠喂以高脂饲料致实验性高脂血症 ,然后分别分组给予硒和/或VE ,4wk后取血及心、肝、肾组织匀浆 ,采用硝酸还原酶等方法测定上述组织的NO和NOS含量。结果 :高脂饲料可致血清、心、肝、肾组织中NO含量及NOS活性降低 ;硒和/或VE能不同程度地增加这些组织中NO含量及心、肝、肾中的NOS活性 (P<0 05或P<0 01) ,且两者合用比单用作用更明显。结论 :硒和/或VE可致实验性高脂血症大鼠心、肝、肾及血清中的NO和NOS发生改变。     

Dinoflagellate polyether within the yessotoxin, pectenotoxin and okadaic acid toxin groups: Characterization, analysis and human health implications

Diarrhetic Shellfish Poisoning (DSP) is a specific type of food poisoning, characterized by severe gastrointestinal illness due to the ingestion of filter feeding bivalves contaminated with a specific suite of toxins. It is known that the problem is worldwide and three chemically different groups of toxins have been historically associated with DSP syndrome: okadaic acid (OA) and dinophysistoxins (DTXs), pectenotoxins (PTXs) and yessotoxins (YTXs). PTXs and YTXs have been considered as DSP toxins because they can be detected with the bioassays used for the toxins of the okadaic acid group, but diarrhegenic effects have only been proven for OA and DTXs. Whereas, some PTXs causes liver necrosis and YTXs damages cardiac muscle after intraperitoneal injection into mice. On the other hand, azaspiracids (AZAs) have never been included in the DSP group, but they cause diarrhoea in humans. This review summarizes the origin, characterization, structure, activity, mechanism of action, clinical symptoms, method for analysis, potential risk, regulation and perspectives of DSP and associated toxins produced by marine dinoflagellates.     

急性心肌梗死患者血糖、糖化血清蛋白和N末端B型钠尿肽原的临床检测价值

目的 通过观察急性心肌梗死(AMI)患者血糖(Glu)、糖化血清蛋白(GSP)、B型钠尿肽原(NT-proBNP)水平变化及并发症发生状况,探讨联合检测的临床价值.方法 据AMI患者入院时有无合并糖尿病分为已知糖尿病组、未知糖尿病组及无糖尿病组,于入院时采集患者外周血,检测血清Glu(葡萄糖氧化酶法)、GSP(果糖胺法)及NT-proBNP(电化学发光免疫分析法)水平,并观察患者住院期间并发症的发生情况.结果 AMI各组患者入院时,已知糖尿病组血清Glu、GSP和NT-proBNP水平[(9.16±0.97) mmol/L、(2.14±0.47) mmol/L和(531.1±99.0) pg/ml]显著高于正常对照组[(5.08±0.91) mmol/L、(1.98±0.41) mmol/L和(47.7±11.6) pg/ml] (P＜0.05);未知糖尿病组血清Glu、GSP和NT-proBNP水平[(13.73±1.39) mmol/L、(3.49±0.62) mmol/L和(966.3±120.7 pg/ml)]显著高于正常对照组(P＜0.05);无糖尿病组血清Glu和NT-proBNP水平[(8.62±0.85) mmol/L和(319.2±87.4) pg/ml]显著高于正常对照组(P＜0.05),但血清GSP水平[(2.06±0.43) mmol/L]差异无统计学意义(P＞0.05);未知糖尿病组与已知糖尿病组及无糖尿病组比较,血清Glu、GSP和NT-proBNP水平差异均具有统计学意义(P＜0.05).住院期间,已知糖尿病组及未知糖尿病组心血管事件发生率(46.7％、64.1％)均显著高于无糖尿病组(28.0％)(x2=3.847、11.600,P＜0.05),未知糖尿病组与已知糖尿病组差异无统计学意义(x2=2.564,P＞ 0.05).结论 血清GSP可作为区分AMI患者Glu增高属于应激性还是糖尿病性的重要依据,血清NT-proBNP可判断AMI患者心功能损害程度.联合检测血清Glu、GSP及NT-proBNP水平,可作为指导AMI患者的临床治疗及预测预后的指标.     

Submicron Size Particles of a Murine Monoclonal Antibody Are More Immunogenic Than Soluble Oligomers or Micron Size Particles Upon Subcutaneous Administration in Mice

Protein aggregates are one of the several risk factors for undesired immunogenicity of biopharmaceuticals. However, it remains unclear which features determine whether aggregates will trigger an unwanted immune response. The aim of this study was to determine the effect of aggregates' size on their relative immunogenicity. A monoclonal murine IgG1 was stressed by exposure to low pH and elevated temperature followed by stirring to obtain aggregates widely differing in size. Aggregate fractions enriched in soluble oligomers, submicron size particles and micron size particles were isolated via centrifugation or size-exclusion chromatography and characterized physicochemically. The secondary and tertiary structures of aggregates were altered in a similar way for all the fractions, while no substantial chemical degradation was observed. Development of anti-drug antibodies was measured after subcutaneous administration of each enriched fraction to BALB/c mice. Among all tested fractions, the most immunogenic was the one highly enriched in submicron size particles (～100-1000 nm). Fractions composed of micron size (>1-100 μm) particles or soluble oligomers (<100 nm) were not immunogenic under the dosing regimen studied in this work. These results show that aggregate size is an important factor for protein immunogenicity.     

Autoimmune response in MRL+/+ mice following treatment with dichloroacetyl chloride or dichloroacetic anhydride

Dichloroacetyl chloride (DCAC) is formed from trichloroethene (TCE), which is implicated in inducing/accelerating autoimmune response. Due to its potent acylating activity, DCAC may convert proteins to neo-antigens and thus could induce autoimmune responses. Dichloroacetic anhydride (DCAA), which is a similar acylating agent, might also induce autoimmune responses. To evaluate if chloroacylation plays a role in the induction of autoimmunity, we have measured the autoimmune responses following treatment with DCAC or DCAA in autoimmune-prone MRL+/+ mice. Five-week-old female mice were injected intraperitoneally (twice weekly) with 0.2 mmol/kg of DCAC or DCAA in corn oil for 6 weeks. Total serum IgG, IgG1, and IgE levels were significantly increased in DCAC-treated mice as compared to controls. These increases corresponded with increases in DCAC-specific IgG and IgG1 levels. Total serum IgM was decreased in both DCAC- and DCAA-treated mice. Antinuclear antibodies, measured as an indication of systemic autoimmune responses, were increased in both DCAC- and DCAA-treated mice. Of eight Th1/Th2 cytokines measured in the serum, only IL-5 was significantly decreased in both treatment groups. The cytokine secretion patterns of splenic lymphocytes after stimulation with antibodies against CD3 (T cell receptor-mediated signal) and CD28 (costimulatory signal) differed between treatment and control groups. Levels of IL-1, IL-3, IL-6, IFN-gamma, G-CSF, and KC were higher in cultures of stimulated splenocytes from either DCAC- or DCAA-treated mice than from controls. The level of IL-17 was only increased in cultures from DCAC-treated mice. Increased lymphocytic populations were found in the red pulp of spleens following treatment with either DCAC or DCAA. In addition, thickening of the alveolar septa in the lungs of DCAC- or DCAA-treated mice was observed. The lung histopathology in exposed mice was consistent with the symptomology observed in welders exposed to DCAC/phosgene. Thickening was more pronounced in DCAC-treated mice. Our data suggest that DCAC and DCAA elicit autoimmune responses in MRL+/+ mice that might be reflective of their chloroacylation potential in vivo.