吗啡染毒大鼠死后体内吗啡的分布研究

作者采用免疫组织化学（PAP）法观察慢性吗啡染毒大鼠死后，在不同时间尸体内吗啡公布的变化。发现随着死后时间的延长，吗啡在大鼠CNS的分布无明显改变；其它内脏器官组织中吗啡分布则有明显改变；内脏脂肪组织原未查见吗啡，随死后时间延长，吗啡含量增加．证实大鼠染毒死后尸体内吗啡存在重分布现象。初步讨论了死后体内吗啡分布的变化与死后时间的关系，为吸毒致死后尸检取材提供依据。     

吗啡类毒品中毒死者主要脏器内吗啡分布的研究

目的研究吗啡在人体脏器中的分布情况,筛选适于免疫组化方法检测吗啡的脏器。方法用免疫组化SP法对8例吗啡类毒品中毒死亡尸体脑、肾、心、肝等脏器中的吗啡进行定位和半定量观察。结果吗啡主要分布在某些实质细胞的胞浆内,肾脏和脑组织的吗啡含量高,不同器官、不同案例吗啡的分布情况差异较大。结论肾脏和脑是实际检案时用免疫组化方法检测吗啡的理想检材。     

吗啡在体内的分布变化研究进展

吗啡在体内的分布变化研究进展李利华，吴家文，江秉康（华西医科大学法医病理教研室；四川成都６１００４１）研究吗啡在体内的分布和变化是药理学及麻醉学中的重要课题。阐明吗啡毒副作用的机理，为临床合理用药提供理论依据。８０年代以来，海洛因及吗啡在世界范围内广...     

急性吗啡中毒大鼠主要器官内吗啡的免疫组化定位研究

一次静脉注射１２．５ｍｇ／ｋｇ。ｂｗ的盐酸吗啡染毒雄性Ｓ－Ｄ大鼠，２小时后处死，取其脑、肾、肝、肺、心组织以２％戊二醛和４％多聚甲醛混合液固定后，常规石蜡切片。运用抗吗啡抗血清及ＳＡＢＣ技术染色。结果显示上述组织切片有不同程度的阳性染色，阳性着色主要见于肾髓质部分肾小管上皮细胞，肝脏中央静脉周围的肝细胞、肺泡上皮细胞及肺内小支气管粘膜上皮细胞、中脑部分神经细胞、室管膜细胞、心肌细胞．以及各器官小血管及毛细血管内皮细胞胞浆、血浆及肾小管腔内尿液。     

急性吗啡中毒大鼠主要脏器内吗啡分布变化的研究

研究急性吗啡中毒大鼠随给药和死后时间延长 ,主要脏器内吗啡的分布变化规律 ,为吗啡类毒品中毒死亡者尸检取材提供依据。采用免疫组织化学SP技术 ,观察 44只尾静脉注射吗啡的大鼠。从给药后 15min到 5h ,从死后即刻至 48h ,脑、肾、心、肝等脏器内吗啡分布的变化规律。结果表明 ,注射吗啡后短时间内各脏器均有吗啡存在 ,主要分布在某些实质细胞的胞浆内 ,且随时间延长吗啡含量上升 ,达高峰 ( 1h)后逐渐减少或消失。不同组织器官的吗啡含量及其变化速率差异巨大。脑内吗啡出现早 ( 15min) ,消失晚 ( 5h) ,峰值高 ,死后衰减慢 ( 4 8h仍呈强阳性 )。肾脏次之 ,但明显优于心、肝。免疫组化SP技术可作为一种鉴定吗啡类毒品中毒的特异性方法 ,脑、肾是其较理想的检材     

大鼠死后心血吗啡浓度变化的HPLC检测

采用高效液相邑谱分析技术（HPLC）检测治疗量及中毒量吗啡肌注大鼠死后心血中吗啡浓度变化。结果表明，以治疗量吗啡肌往大鼠，在死后96h内，心血中吗啡浓度随死后时间增加而显著升高（P＜0．01），吗啡浓度水平与死后时间里显著正相关；以中毒量吗啡肌注大鼠，在死后12h内，心血吗啡浓度无明显变化；死后24h、48h及96h，随死后时间延长，心血中吗啡浓度逐渐升高（P＞0．01），其递增强度不如治疗量吗啡组大鼠的明显.本研究证实，死后尸体心血吗啡浓度明显受生前剂量的影响，且在死后96h内，随死后时间增加．心血中吗啡浓度少数不断增高。     

毛发中吗啡放射免疫分析

毛发作为药物分析检材，与体液检材所不同的是，具有容易获得，稳定和保留药物时间长等特点。吗啡是鸦片类物质在机体内的代谢产物，不仅存在于血液和尿液中，同时也蓄积在毛发中。由于体液中的吗啡停留时间短，只能提供短期的用药信息；而毛发中的吗啡是随用药时间延续，长期积累所致。因此，毛发分析避免了暂时药物切断和掺假的干扰，是判断药物滥用的行之有效的手段，是对体液分析的补充［’｝。本文采用本室建立的吗啡放射免疫分析测定方法（习，对鸦片依赖者和正常人（非依赖者）毛发进行比对测定。结果证明，用放射免疫分析毛发中吗啡…     

吗啡依赖和戒断大鼠中枢cAMP和cGMP含量的变化

目的　研究吗啡依赖和催促戒断大鼠中枢cAMP和cGMP含量变化。方法　以剂量递增法连续皮下注射吗啡建立吗啡依赖模型 ,采用放射免疫学方法检测脑内cAMP和cGMP含量。结果　与生理盐水对照组大鼠比较 ,吗啡依赖大鼠的纹状体、间脑、中脑、脑桥和海马内cGMP含量均显著降低 ,cAMP含量和cAMP/cGMP比值显著升高 ,而小脑则无类似变化 ;与吗啡依赖组大鼠比较 ,纳洛酮催促戒断大鼠 ,其海马和纹状体内的cGMP含量显著下降 ,cAMP含量和cAMP/cGMP比值显著升高 ;其它部位无明显变化。结论　中枢cAMP和cGMP含量变化 ,可能是形成和维持吗啡依赖和戒断的重要分子机制之一。     

大鼠死后心血中吗啡浓度变化的HPLC检测

本文采用高效液相色谱（HPLC）分析技术检测了治疗量及中毒量吗啡肌注大鼠死后心血中吗啡浓度变化．结果表明．以治疗量吗啡肌注大鼠死后96h内，心血中吗啡浓度随死后时间增加而显著升高（P<0.01），吗啡浓度水平与死后时间呈显著正相关．以中毒量吗啡肌注大鼠死后12h内，心血中吗啡浓度无明显变化．死后24h、48h及96h．随死后时间延长，心血中吗啡浓度逐渐升高（P＞0.01），其递增强度不如治疗量吗啡组大鼠的明显．本研究证实死后尸体心血吗啡浓度明显受生前剂量的影响，且死后96h内，随死后时间延长心血中吗啡浓度不断增高．本文初步探讨了死后心血吗啡浓度变化发生的可能机制．为海洛因或吗啡中毒死亡的血液检测结果评判及死因分析提供理论依据．     

氯胺酮、甲基苯丙胺和吗啡金标单抗试剂盒的研制

目的建立同步检测氯胺酮、甲基苯丙胺和吗啡的方法。方法将胶体金标记的抗氯胺酮、抗甲基苯丙胺和抗吗啡单克隆抗体浸涂在玻璃纤维膜上，将氯胺酮、甲基苯丙胺和吗啡的完全抗原以及羊抗鼠多克隆抗体喷涂在硝酸纤维素膜上，分别标定为检测区（T）和质控区（C）。样本中游离的氯胺酮、甲基苯丙胺和吗啡分别与包被的完全抗原免疫竞争结合胶体金标记抗氯胺酮、抗甲基苯丙胺和抗吗啡单克隆抗体。以质控区和检测区是否出现紫红色条带判读结果。结果对66种药品和毒品的特异性测试表明，该试剂盒仅识别氯胺酮及其代谢物、甲基苯丙胺及其衍生物和吗啡类；对人体尿样中的氯胺酮、甲基苯丙胺和吗啡检测阈值分别为1000ng／ml、1000ng／ml和300ng／ml；与GC／MS对照试验结果一致；试剂盒稳定性较好，在常温下可较长时间保存。结论本文研制的试剂盒可用于样本中氯胺酮、甲基苯丙胺和吗啡成分定性的同步检测。     

Postmortem disposition of morphine in rats

The antemortem and postmortem distribution of morphine was studied in rats for the purpose of establishing whether drug distribution is altered after death. Samples were examined for free and total morphine concentration, pH and water content at 0-96 h after death. Morphine was administered antemortem at various intervals. All groups of rats studied showed a significant (P less than 0.05) increase in postmortem cardiac blood morphine concentrations. These changes, which are detectable within 5 min after death are likely to be related to an observed, rapid decrease in cardiac blood pH from 7.34 +/- 0.02 to 6.74 +/- 0.05. Significant increases in free morphine levels were, also, observed 24 and 96 h after death in liver, heart and forebrain while urine morphine levels decreased. The liver showed the greatest increase (20-fold) in free morphine levels 96 h after death, while hindbrain levels did not significantly change. Bacterial hydrolysis of morphine glucuronides accounted only in part for the observed increase in free morphine concentration. Postmortem fluid movement and pH-dependent drug partitioning was detected. It would appear that several mechanisms are responsible for postmortem drug distribution. Understanding the mechanisms and patterns responsible may eventually lead to better choices of postmortem tissue which may better represent antemortem drug levels.     

酶水解后同时分析海洛因、吗啡、可待因

本文以家兔为模型，用酶水解、丙酸酐衍生化处理吸毒后的尿样，用ＧＣ／ＭＳ同时分析海洛因、吗啡、可待因。     

大鼠肥大细胞的法医学实验研究──过氧化物酶活性变化、脱颗粒超微结构改变及分布

采用植物血凝素（PHA）致敏S、D大鼠，2周后处死，灌洗腹腔，用阿拉伯胶分离液分离肥大细胞（MC），将PHA与MC在37C水治中孵育，攻击MC使之受损伤，于伤后5、10、15、20、30和60分钟，分别吸取孵有液作聚丙烯酰胺凝胶圆盘电泳，分离MC过氧化物酶；用抗坏血酸-联苯胺染色显带，并计算酶的相对活性。另取20分钟时孵育液，常规固定，包埋，切片及染色后用透射电镜观察MC脱颗粒叶的形态。用常规石蜡切片，甲苯胺蓝染色，对SD大鼠的35种组织中MC的分布进行光镜观察。结果发现；（1）大鼠腹腔MC显示-条过氧化物酶电泳区带；（2）MC经PHA攻击受损伤10分钟内，过氧化物酶相对活性升高不明显；15、ZO、3O分钟时实验组酶活性升高，与对照组的差异明显；到60分钟时两组酶活性无显著差异；（3）MC与PHA孵育20分钟时，脱颗粒率为76％，MC浆内可见有膜性和无膜性两种类型的空泡．对照组大鼠MC脱颗粒仅为16％；（4）大鼠皮肤、肺、呼吸道、消化道、骨髓及胸腺MC含量较多；泌尿生殖道次之；心、肝、脾、肾等组织较少；脑、脊髓、脑垂体、肾上腺及睾丸等组织中未见MC，从而为法医损伤时间的推测提供方法与依据。     

Morphine and 6-acetylmorphine concentrations in blood, brain, spinal cord, bone marrow and bone after lethal acute or chronic diacetylmorphine administration to mice

The aim of this study was to evaluate postmortem incorporation of opiates in bone and bone marrow after diacetylmorphine (heroin) administration to mice. Mice were given acute (lethal dose of 300 mg/kg) or chronic (10 and 20 mg/kg/24 h for 20 days) intraperitoneal administration of diacetylmorphine. The two metabolites of diacetylmorphine, 6-acetylmorphine (6-AM) and morphine, were extracted from whole blood, brain, spinal cord, bone marrow and bone (after hydrolysis) using a liquid/liquid method. Quantification was performed by gas chromatography-mass spectrometry (GC/MS). Results showed that after acute administration, opiates were present in all studied tissues. Morphine concentrations appeared to be higher than those of 6-AM in blood (52.4 microg/mL versus 27.7 microg/mL, n=12), bone marrow (87.8 ng/mg versus 8.9 ng/mg, n=6) and bone (0.85 ng/mg versus 0.43 ng/mg, n=6), but 6-AM concentrations were higher than those of morphine in brain (14.0 ng/mg versus 7.4 ng/mg, n=12) and spinal cord (27.8 ng/mg versus 20.8 ng/mg, n=12). No correlation was found for both compounds between blood concentrations and either brain, spinal cord, bone or bone marrow concentrations while a significant one was found between brain and spinal cord concentrations either for morphine (r=0.89, n=12, p<0.001) or 6-AM (r=0.93, n=12, p<0.001), the concentration being higher in spinal cord than in brain. When bones were stored for 2 months, only 6-AM remained in bone marrow but not in bone. After chronic administration, mice being sacrificed by cervical dislocation 24 h after the last injection, no opiate was detected in any studied tissues. Further studies are required, in particular in human bones, but these results seem to show that 6-AM could be detect in bone marrow several weeks after the death and could be an alternative tissue for forensic toxicologist to detect a fatal diacetylmorphine overdose, even if no correlation between blood and bone marrow was observed. On the other hand, neither bone tissue nor bone marrow will allow the confirmation of a chronic diacetylmorphine use.     

Distribution of drugs of abuse within specific regions of the human brain

Since concentration of drugs of abuse found in the brain better reflect drug concentration at their site of action, brain specimens are useful in the determination of the role of drugs of abuse in the cause of death. In order to allow for the routine use of brain specimens in this field, a comprehensive database with reliable reference values is needed and should include both post-mortem data for cases where drugs have been taken in therapeutic doses as well as for cases of overdose. In this study, a semi-automated extraction procedure, in combination with gas chromatography/mass spectrometry (GC–MS) using stable isotope internal standards was applied to yield reproducible, quantitative results which could be used to investigate the distribution patterns of drugs of abuse within specific regions of the brain, by analyzing several segments of both medulla oblongata and cerebellum. A homogenous distribution of unconjugated morphine, dihydrocodeine, and benzoylecgonine within the investigated segments of medulla oblongata or cerebellum could be found. However, when these two brain regions from the same case were compared to each other, significantly higher concentrations of unconjugated morphine, dihydrocodeine, and benzoylecgonine were found in the cerebellum than in the medulla oblongata.     

Distribution of morphine and morphine glucuronides in body tissue and fluids--postmortem findings in brief survival

An intoxication following administration of morphine, tramadol and atracurium in a suicide case is reported. The route of administration and the amount of the particular drug were known from the investigation of the death scene and the findings of the postmortem examination. Tramadol was present in the gastric contents as well as in blood, liver, kidney and brain samples, whereas the drug could not be detected in muscle. All body fluids and tissues investigated contained morphine as well as its 3- and 6-glucuronides with the exception of muscle tissue. The concentrations of morphine and its glucuronide metabolites were determined by LC/MS following solid phase extraction. Interestingly, the concentration of M6G in brain, liver and kidney were close to the concentration of M3G in the particular tissue. This phenomenon might be explained by a preferential hydrolysis of M3G or by a preferential formation of M6G postmortem. Measurement of morphine and M6G in femoral blood and cerebrospinal fluid may be a useful indicator in rapid deaths.