Validation of a HPLC/MS method for simultaneous quantification of clonidine,morphine and its metabolites in human plasma

A high‐performance liquid chromatography–tandem mass spectrometry method was developed and validated for the simultaneous quantification of morphine, morphine's major metabolites morphine‐3‐glucuronide and morphine‐6‐glucuronide, and clonidine, to support the pharmacokinetic analysis of an ongoing double‐blinded randomized clinical trial that compares the use of morphine and clonidine in infants diagnosed with neonatal abstinence syndrome. Plasma samples were processed by solid‐phase extraction and separated on an Inertsil ODS‐3 (4 μm) column using an 0.1% formic acid in water–0.1% formic acid in methanol gradient. Detection of the analytes was conducted in the positive multiple reaction monitoring mode. The range of quantitation was 1–1000 ng/mL for morphine, morphine‐3‐glucuronide and morphine‐6‐glucuronide, and 0.25–100 ng/mL for clonidine. Intra‐day and inter‐day accuracy and precision were ≤15% for all analytes across the quantitation range. Extraction recovery rates were ≥94% for morphine, ≥90% for M3G, ≥87% for M6G and ≥ 79% for clonidine. Matrix effect ranged from 85–94% for clonidine to 101–106% for M3G. The method fulfilled all predetermined acceptance criteria and required only 100 μL of starting plasma volume. Furthermore, it was successfully applied to 30 clinical trial plasma samples.     

Neuroprotection of N-benzyl Eicosapentaenamide in Neonatal Mice Following Hypoxic–Ischemic Brain Injury

Maca (Lepidium meyenii) has emerged as a popular functional plant food because of its medicinal properties and nutritional value. Macamides, as the exclusively active ingredients found in maca, are a unique series of non-polar, long-chain fatty acid N-benzylamides with multiple bioactivities such as antifatigue characteristics and improving reproductive health. In this study, a new kind of macamide, N-benzyl eicosapentaenamide (NB-EPA), was identified from maca. We further explore its potential neuroprotective role in hypoxic–ischemic brain injury. Our findings indicated that treatment with biosynthesized NB-EPA significantly alleviates the size of cerebral infarction and improves neurobehavioral disorders after hypoxic–ischemic brain damage in neonatal mice. NB-EPA inhibited the apoptosis of neuronal cells after ischemic challenge. NB-EPA improved neuronal cell survival and proliferation through the activation of phosphorylated AKT signaling. Of note, the protective property of NB-EPA against ischemic neuronal damage was dependent on suppression of the p53–PUMA pathway. Taken together, these findings suggest that NB-EPA may represent a new neuroprotectant for newborns with hypoxic–ischemic encephalopathy.     

重组人硫氧还蛋白对大鼠内毒素肺损伤保护作用

目的:探讨重组人硫氧还蛋白1(rh Trx-1)对新生SD大鼠急性内毒素肺损伤的保护作用.方法:制备、鉴定rh Trx-1;将新生SD大鼠分为对照组、脂多糖(LPS)实验组和LPS+rh Trx-1干预组.腹腔注射LPS(质量分数为5 mg/kg),制备急性内毒素肺损伤模型,干预组于LPS注射前30 min腹腔注射rh Trx-1(质量分数为10 mg/kg),观察注射后24 h各组新生SD大鼠的一般情况以及死亡率;重复实验,每组分别于实验后的2、8 h随机处死大鼠,提取肺组织用于病理切片观察.结果:(1)成功制备rh Trx-1;(2)LPS组大鼠表现出活动少,反应差,精神萎靡,毛发无光泽,口周发绀等全身炎症反应症状,濒死状态时表现为毛色青灰,呼吸浅表;LPS+h Trx-1干预组动物症状明显减轻;阴性对照组、LPS实验组和LPS+rh Trx-1干预组大鼠24 h死亡率分别为0%,67%、17%(2=14.400,P=0.001);(3)LPS组肺组织损伤严重,可见肺组织结构不完整,肺泡间隔明显增宽,肺泡腔内有红细胞、炎症细胞及浆液渗出,血管周围组织可见白细胞浸润,毛细血管有明显充血及扩张表现,随时间延长有加重趋势,LPS+rh Trx-1干预组病理改变较内毒素组明显减轻,肺泡结构尚正常,轻度肺水肿,肺泡间隔稍增宽,红细胞渗出明显减少,肺泡及肺间质中白细胞浸润减少.结论:重组人硫氧还蛋白1对急性内毒素肺损伤的新生大鼠具有保护作用.     

小麦-混合蛋白日粮中色氨酸含量对仔猪生长性能的影响

在小麦—混合蛋白日粮中使用0.16%,0.18%,0.20%,0.22%的色氨酸水平饲喂仔猪45 d,结果显示,在0.18%和0.20%色氨酸水平下仔猪平均日增重和料重比极显著提高(P<0.01),当色氨酸水平达到0.22%后,生产性能有所降低;其中0.20%色氨酸含量的饲喂效果最好,仔猪日增重达547.78g±47.99g,比对照组高16.10%,料重比3.21±0.79,比对照组低25.98%,血清尿素氮随色氨酸含量减少。     

如何提高病毒外源因子检测中小鼠和乳鼠检查法的成功率

随着生物制品在临床上的应用日益广泛,其安全性也备受关注。根据国家规定,拟上市的生物制品需依照2015版药典要求进行病毒外源因子检测以控制其质量,但其中较为关键的小鼠、乳鼠检查法在实际操作中存在一定的困难,导致检测结果易出现假阴性或假阳性,有鉴于此,本文以广东省医学实验动物中心丰富的检测经验为依据,对小鼠及乳鼠检查法的关键点进行归纳总结,拟提供高效实用的操作方法并提高该检查法的成功率。     

基于不相关局部敏感鉴别分析的新生儿疼痛表情识别

针对新生儿疼痛表情识别中的特征提取问题,提出一种不相关局部敏感鉴别分析（Uncorrelated Locality Sensitive Discriminant Analysis,ULSDA）算法.首先,在局部敏感鉴别分析（LSDA）算法的基础上,通过附加投影向量正交性的约束条件,定义了ULSDA的目标函数;然后,推导出求解特征子空间正交投影向量的迭代公式;最后,将输入的高维图像数据投影到这个特征子空间,求出特征向量.ULSDA算法不仅继承了LSDA算法所具有的有监督、局部流形结构保持等特性,而且消除了LSDA算法所提取出的鉴别特征的相关性,从而增强了特征的鉴别能力,比LSDA算法具有更好的分类识别能力.在新生儿表情图像库上的疼痛表情识别实验结果表明,提出的ULSDA方法是有效可行的.当每类表情的训练样本图像为150幅时,采用ULSDA算法获得的平均识别率达到了82.07％,优于主成分分析（PCA）、线性鉴别分析（LDA）、局部敏感鉴别分析（LSDA）等特征提取方法.     

复合微生态制剂对断奶仔猪生长性能和粪便微生物的影响

旨在探讨复合微生态制剂对断奶仔猪生长性能、粪便微生物的影响．选用90头30-32日龄、体重相近的（长×大）断奶仔猪,将其分为3个处理组,每组3个重复,每个重复10头猪（5公5母）．3个处理组分别是对照组（只饲喂基础日粮）、抗生素组（基础日粮＋抗生素）和复合微生态制剂组（基础日粮＋复合微生态制剂）：试验期30d．结果表明：试验各个阶段和整个试验期,饲料中添加复合微生态制剂对断奶仔猪生长性能无显著影响（P〉0．05）．试验结束时,饲喂复合微生态制剂的断奶仔猪的平均日增重分别比基础日粮组和抗生素组提高5．63％和7．73％、料重比分别下降7．24％和2．82％;试验第15d和第30d,饲料中添加复合微生态制剂对断奶仔猪粪便中乳酸杆菌数无显著影响（P〉0．05）;试验第30d,饲喂复合微生态制剂的仔猪粪便大肠杆菌的数量显著低于对照组和抗生素组（P〈0．05）．综合分析,复合微生态制剂可以抑制粪便中大肠杆菌的生长,促进断奶仔猪健康,改善其生长性能．     

鱼腥草与TMP对仔猪黄痢病原菌的联合抑菌作用

利用鱼腥草与TMP联合用药进行仔猪黄痢病原菌的体外抑菌试验,探讨中西药结合的抗菌效果。结果表明,鱼腥草和TMP对致病性大肠杆菌的最小抑菌浓度分别为1．25g／mL和1．25mg／mL;二者联合用药后,鱼腥草和TMP的最小抑菌浓度分别下降至0．625g／mL和0．625mg／mL,抗菌作用表现为相加作用,抑菌效果显著。     

Low n-6/n-3 Gestation and Lactation Diets Influence Early Performance,Muscle and Adipose Polyunsaturated Fatty Acid Content and Deposition,and Relative Abundance of Proteins in Suckling Piglets

Elevated omega-6 (n-6) and omega-3 (n-3) polyunsaturated fatty acids (PUFAs) ratios in swine diets can potentially impose a higher risk of inflammatory and metabolic diseases in swine. A low ratio between the two omega PUFAs has beneficial effects on sows’ and piglets’ production performance and immunity status. At present, there are few studies on how sow nutrition directly affects the protein and fat deposition in suckling piglets. Two groups of sows were fed diets with high or low n-6/n-3 polyunsaturated ratios of 13:1 (SOY) and 4:1 (LIN), respectively, during gestation and lactation. Longissimus dorsi muscle and adipose tissue from newborn piglets, nourished only with sow’s milk, were subjected to fatty acid profiling by gas chromatography–mass spectrometry (GC-MS) and to proteomics assays based on nano-liquid chromatography coupled to high-resolution tandem mass spectrometry (nLC-HRMS). Fatty acid profiles on both muscle and adipose tissues resembled the magnitude of the differences between fatty acid across diets. Proteomic analysis revealed overabundance of 4 muscle and 11 adipose tissue proteins in SOY compared to LIN in both piglet tissues. The detected overabundance of haptoglobin, an acute-phase protein, and the stimulation of protein-coding genes and proteins related to the innate immune response and acute inflammatory response could be associated with the pro-inflammatory role of n-6 PUFAs.     

机械通气治疗新生儿肺出血35例临床分析

目的:总结机械通气在新生儿肺出血治疗中的临床应用,为新生儿肺出血救治提供指导。方法:对2003年6月至2008年1月我院收治的35例新生儿肺出血,应用机械通气的治疗方法进行总结分析。结果:35例中治愈30例,治愈率为85.71%,死亡5例,病死率为14.29%。结论:机械通气是一种目前治疗新生儿肺出血不可替代的治疗方法。