可卡因对小鼠离体脾细胞功能的影响

目的 观察盐酸可卡因对小鼠体外培养脾细胞功能的影响.方法 采用小鼠脾细胞体外培养的方法,在细胞培养液中加入终质量浓度分别为10、20和100μg/mL的盐酸可卡因,连续培养.检测脾细胞的ConA增殖反应、IL-2、IFN-γ的活性.结果 实验组ConA诱导的小鼠脾细胞增殖反应明显降低,IL-2和IFN-γ的活性显著下降,与对照组相比较有显著性差异(P＜0.05;P ＜0.01).结论 可卡因对体外培养脾细胞功能有显著的抑制作用.     

可卡因染毒致小鼠免疫功能损伤的影响

目的考察可卡因染毒对小鼠免疫功能的影响。方法 615纯系雄性小鼠20只,随机均分为实验组和对照组;实验组小鼠以30mg/kg盐酸可卡因每天1次进行腹腔连续注射,空白组给予相同量盐水同法注射盐水;第30d处死小鼠,测量体重与脏器系数,用四甲基偶氮唑蓝(MTT)法检测脾细胞、胸腺细胞与腹腔巨噬细胞(PMφs)的增殖活性;用生物活性检测法检测IL-1与IL-2的活性;用ELISA法检测IFN-γ与TNF-α含量。结果实验组小鼠脾细胞、胸腺细胞与PMφs的增殖反应明显降低;脾细胞培养上清液中IL-2与IFN-γ活性明显降低;PMφs培养上清中IL-1与TNF-α活性明显降低;体重、脾脏与胸腺系数也明显降低;上述指标较之对照组均有显著性差异(P0.01)。结论在活体条件下可卡因染毒对小鼠免疫系统功能有明显抑制作用。     

可卡因染毒致小鼠免疫功能损伤的影响

目的考察可卡因染毒对小鼠免疫功能的影响。方法615纯系雄性小鼠20只,随机均分为实验组和对照组;实验组小鼠以30mg／kg盐酸可卡因每天1次进行腹腔连续注射,空白组给予相同量盐水同法注射盐水;第30d处死小鼠,测量体重与脏器系数,用四甲基偶氮唑蓝（MTr）法检测脾细胞、胸腺细胞与腹腔巨噬细胞（PMφs）的增殖活性;用生物活性检测法检测IL-1与IL-2的活性;用ELISA法检测IFN-γ／与TNF—α含量。结果实验组小鼠脾细胞、胸腺细胞与PMφs的增殖反应明显降低;脾细胞培养上清液中IL-2与IFN-γ活性明显降低;PMφs培养上清中IL-1与TNF—α活性明显降低;体重、脾脏与胸腺系数也明显降低;上述指标较之对照组均有显著性差异（P〈0．01）。结论在活体条件下可卡因染毒对小鼠免疫系统功能有明显抑制作用。     

可卡因对小鼠离体腹腔巨噬细胞功能的影响

目的研究可卡因对体外培养小鼠腹腔巨噬细胞功能的影响。方法无菌分离小鼠腹腔巨噬细胞,制备单细胞悬液,分别用不同浓度的可卡因作用于正常的和LPS活化的腹腔巨噬细胞。MTT法检测其增殖反应;ELISA法检测巨噬细胞培养上清中TNF-α的分泌水平;生物活性检测法检测IL-1的活性。结果可卡因在终浓度10、20和100μtg／mL时能明显抑制LPS刺激的腹腔巨噬细胞增殖反应,减少TNF-α、IL．1的释放,与对照组相比较有显著性差异（P〈0．01）。结论可卡因对体外培养腹腔巨噬细胞功能有明显抑制作用。     

甲基苯丙胺对大鼠认知和脑电图的影响

目的研究甲基苯丙胺（methamphetamine,MA）对脑功能的影响机制,实验设计观察慢性反复使用MA是否同时对大鼠认知行为和脑电活动产生影响。方法 14只雄性SD大鼠（180～200g）随机分为生理盐水注射组（对照组）和MA注射组（实验组5mg/kg体重）每天一次腹腔注射。连续注射7d后进行被动避暗回避行为测试和脑电图检测。结果与对照组相比,实验组大鼠表现为被动避暗回避行为的潜伏期延长,脑电图频谱分析显示δ波相对功率值减小,β波相对功率值增大,差异有统计学意义（P〈0.05）。结论慢性反复使用MA对大鼠被动回避行为和脑电活动均有影响,说明MA滥用造成的认知行为改变与中枢神经系统电生理活性改变有关。     

人体中可卡因代谢物：尿液中四种未报道过的可卡因代谢物的鉴定

在体内,可卡因由于酶的作用和化学过程迅速转变为代谢物,因此通常尿中仅能提出很少可卡因。尿中两种主要代谢物为苯甲酰牙子碱及牙子碱甲基酯。Amber 最近报道可卡因在尿中14～17%以苯甲酰牙子碱形式被提出,12～21%以牙子碱甲基酯形式被提出。另外报道的体内尿中可卡因代谢物包括:降可卡因、牙子碱、苯甲酰降牙子碱、脱水芽子碱甲基酯、苯甲酰环上羟基化的可卡因、羟基、甲氧基取代的苯甲酰芽子碱。本文报道一种扩展的GC/MS 方法,用于分析药物试验中收集的1例尿样,该     

特异性可卡因核酸适体测试条的研制

目的筛选识别可卡因的特异性核酸适体,制备可卡因核酸适体测试条（盒）。方法通过SELEX技术筛选可卡因核酸适体,制备胶体金标记可卡因完全抗原（COC-BSA）,制作可卡因层析测试条,并对方法特异性、灵敏度、准确性等性能指标进行考察和验证。结果甲基苯丙胺等64种药品和毒品采用本文研制的可卡因测试条检测,结果均为阴性,无交叉反应;检测可卡因最低浓度在5ng/mL,比胶体金单克隆抗体方法检测高5倍;将测试条密封45℃烘烤1个月后,检测结果与未烘烤过的测试条检测结果一致;与GC/MS检测结果对照,准确性为100%。结论本文研制的可卡因核酸适体检测试条,性能稳定,特异性良好,灵敏性和准确性均可达到实际检测的要求,在毒品、毒物的检测中具有广阔的应用前景。     

急性乙醇中毒大鼠脑皮质α-syn表达与神经细胞凋亡的关系

目的观察急性乙醇中毒大鼠脑皮质α-突触核蛋白(α-synuclein,α-syn)表达及神经细胞凋亡的变化,探讨乙醇对神经细胞损害的机制。方法用50%乙醇溶液灌胃方法制作大鼠急性乙醇中毒模型。采用免疫组织化学染色技术和图像分析方法观察大鼠急性乙醇中毒后1 h、3 h、6 h、12 h脑皮质内α-syn和caspase-3的表达。测定阳性细胞数和平均光密度值,分析其变化趋势,并进行方差分析和t检验。结果急性乙醇中毒组大鼠脑皮质α-syn染色阳性细胞数和平均光密度值均呈上升趋势,6 h达到峰值,12 h略有下降,但仍高于1 h、3 h组。中毒后12 h以内,大鼠脑皮质的caspase-3阳性细胞数和平均光密度值均呈逐渐上升趋势。结论急性乙醇中毒后脑水肿、缺氧引起α-syn异常聚集可能参与乙醇中毒后早期神经细胞凋亡过程。     

LC-MS／MS测定尿液中可卡因及其代谢物苯甲酰爱康宁

目的建立尿液中可卡因(cocaine,COC)及其代谢物苯甲酰爱康宁(benzoylecgonine,BZE)的液相色谱-串联质谱分析方法。方法尿液经固相萃取后,用AllurePFP丙基柱分离,以V(甲醇):V(20mmol/L乙酸胺和0.1%甲酸的缓冲溶液)=80∶20为流动相,采用二级质谱多反应监测模式检测COC和BZE。按10mg/kg的剂量对豚鼠腹腔注射可卡因,给药后收集7d尿液。结果尿液中COC和BZE在2.0～100ng/mL质量浓度范围内线性关系良好(r=0.9995),最低检测限(LOD)为0.5ng/mL;回收率大于90%;日内和日间精密度均小于6%;豚鼠尿液中主要检测目标物是BZE,且BZE检测时限也较COC长。结论所建方法灵敏度高,选择性好,适用于尿液中可卡因和苯甲酰爱康宁的检测。     

可卡因及其代谢物苯甲酰爱康宁胶体金检测试剂盒的研制

目的根据胶体金免疫层析的原理,建立一种可以在现场快速检测可卡因及其代谢物苯甲酰爱康宁的方法。方法将胶体金标记的抗可卡因单克隆抗体均匀浸涂在玻璃纤维膜上,用点膜仪将可卡因的完全抗原以及羊抗鼠多抗在硝酸纤维素膜上划线,分别作为可卡因的检测区（T）和质控区（C）。并且对样品垫用0.2%Tween 20和0.5%PVA封闭处理。样本（尿样、唾液、血样等）中游离的可卡因及其代谢物同包被的完全抗原分别免疫竞争结合胶体金标记的抗可卡因单抗。肉眼观察试剂盒的质控区和检测区上有无紫红色带来判读结果。结果可卡因胶体金检测试剂盒对65种毒品、药物的特异性测试中仅识别可卡因及其代谢物。对人体尿样中的可卡因和苯甲酰爱康宁检测阈值均为300ng/mL。与GC/MS对照试验,其准确性分别为98.9%和98.6%。在常温下能稳定保存2年。结论该可卡因检测试剂盒在5min内可以检测出样本中的可卡因及其代谢物成分。     

Monitoring of Levamisole Concentration in Serum of Traffic Participants after Cocaine Consumption

This study highlights the problem of levamisole‐adulterated cocaine in context of active traffic participation. For the purposes of levamisole concentration monitoring in human serum, an analytical method based on LC‐MS/MS and solid‐phase extraction was applied. A Luna 5 μm C18 (2) 100 A, 150 mm × 2 mm column and a mobile phase consisting of A (H₂O/methanol = 95/5, v/v) and B (H₂O/methanol = 3/97, v/v), both with 10 mM ammonium acetate and with 0.1% acetic acid (pH = 3.2), were used. The validation experiments demonstrated that the method applied was appropriate for levamisole quantification in human serum. For 23% of levamisole‐positive samples, the concentrations exceeded 20 ng/mL. Therefore, the interaction of this drug with cocaine has to be considered as important for active traffic participation. As a consequence, monitoring of levamisole concentration in human serum is recommended, as long as it is used as cocaine adulterant.     

Extensive Necrotic Purpura in Levamisole‐Adulterated Cocaine Abuse – A Case Report

Levamisole, which is used as an adulterated compound of cocaine, is currently being seen year after year in cocaine intoxication. For a few cases in the last decade, necrotic purpura and neutropenia after levamisole/cocaine intoxication have been described in the medical community. Herein, we present an original case of levamisole intoxication of a 40‐year‐old woman who smoked heroin and cocaine few during a month. She rapidly presented an extensive necrotic purpura of the nose, cheeks and extremities (lower and upper), and immunologic reactions (positive anti‐MPO and anti‐HNE). Levamisole was detected on hairs with ultra‐high performance liquid chromatography and tandem mass spectrometry. The case reports also a probable cocaine supplier deceit, which bring pure drug for hospital investigation after the intoxication of his client. The intoxicated woman had survived with several skin and chronic pain complications. That case recalls the knowledge about levamisole with a short review of the forensic literature.     

Simultaneous Quantitative Determination of Amphetamines,Opiates, Ketamine,Cocaine and Metabolites in Human Hair: Application to Forensic Cases of Drug Abuse

A method using liquid chromatography–tandem mass spectrometry (LC–MS/MS) to simultaneously quantify amphetamines, opiates, ketamine, cocaine, and metabolites in human hair is described. Hair samples (50 mg) were extracted with methanol utilizing cryogenic grinding. Calibration curves for all the analytes were established in the concentration range 0.05–10 ng/mg. The recoveries were above 72%, except for AMP at the limit of quantification (LOQ), which was 48%. The accuracies were within ±20% at the LOQ (0.05 ng/mg) and between −11% and 13.3% at 0.3 and 9.5 ng/mg, respectively. The intraday and interday precisions were within 19.6% and 19.8%, respectively. A proficiency test was applied to the validated method with z-scores within ±2, demonstrating the accuracy of the method for the determination of drugs of abuse in the hair of individuals suspected of abusing drugs. The hair concentration ranges, means, and medians are summarized for abused drugs in 158 authentic cases.     

An Unusually High Cocaine Blood Concentration in an Impaired Driving Investigation

Cocaine is an illicit drug frequently encountered by forensic practitioners in driving under the influence of drugs (DUID) casework. Whole blood collected from a suspected drugged driver was found to contain 3.000 mg/L cocaine, 1.600 mg/L benzoylecgonine, and 0.260 mg/L methamphetamine. The high concentration of cocaine, while common in overdose death investigations, is unusual for an impaired driving case. Information from the officer revealed that the motorist swallowed cocaine during the traffic stop. Although a cocaine DUID charge could not be pursued, the blood methamphetamine concentration exceeded the State of Nevada “per se” limit for operating a motor vehicle. The motorist was successfully prosecuted for DUID based on his admission of using methamphetamine prior to driving and the blood methamphetamine result. This case highlights the importance of considering case history when interpreting laboratory results and the application of jurisdictional statutes as an approach to prosecuting suspected drug‐impaired drivers.     

Relationships between concentrations of cocaine and its hydrolysates in peripheral blood, heart blood, vitreous humor and urine

Cocaine is known to degrade in vivo and in vitro by several hydrolytic mechanisms. A previous study found that the initial amount of cocaine added to plasma could be accounted for by summing the molar concentrations of cocaine's hydrolysis products and the cocaine remaining after hydrolysis. The present study was undertaken to investigate whether or not relationships might exist between such molar concentration sums for different postmortem bodily fluids. Determinations of cocaine, benzoylecgonine, ecgonine methyl ester, and ecgonine were performed using liquid chromatography/mass spectrometry (LC/MS/MS) with heart blood, femoral blood, vitreous humor (VH), and urine (UR). The results demonstrate a strong correlation between blood and VH concentrations (correlation coefficients of 0.88-0.94), weak correlation between the UR and blood concentrations (correlation coefficients of 0.61-0.64), and weak correlation between UR and VH concentrations (correlation coefficient of 0.59). The results demonstrate that ecgonine is a significant hydrolysate with concentrations on the same order of magnitude as benzoylecgonine. The results are consistent with rapid distribution of the parent drug and its hydrolysates in the blood and VH. The strong correlation between the blood and VH demonstrates that VH is an important medium for toxicology testing when attempting to make a determination of cocaine intoxication.     

溴敌隆中毒研究进展

溴敌隆为二代香豆素类杀鼠药,由其导致的中毒案事件时有发生.本文就溴敌隆的中毒原因、毒理作用、药代动力学以及中毒临床表现、尸体解剖情况、理化检测进行综述.希望能为相关研究和实际检案提供参考.     

Identification of Different Forms of Cocaine and Substances Used in Adulteration Using Near‐infrared Raman Spectroscopy and Infrared Absorption Spectroscopy

Identification of cocaine and subsequent quantification immediately after seizure are problems for the police in developing countries such as Brazil. This work proposes a comparison between the Raman and FT‐IR techniques as methods to identify cocaine, the adulterants used to increase volume, and possible degradation products in samples seized by the police. Near‐infrared Raman spectra (785 nm excitation, 10 sec exposure time) and FT‐IR‐ATR spectra were obtained from different samples of street cocaine and some substances commonly used as adulterants. Freebase powder, hydrochloride powder, and crack rock can be distinguished by both Raman and FT‐IR spectroscopies, revealing differences in their chemical structure. Most of the samples showed characteristic peaks of degradation products such as benzoylecgonine and benzoic acid, and some presented evidence of adulteration with aluminum sulfate and sodium carbonate. Raman spectroscopy is better than FT‐IR for identifying benzoic acid and inorganic adulterants in cocaine.