Hair analysis to document non-fatal pesticide intoxication cases

We reported two non-fatal cases of intoxication with pesticides namely alachlor and carbofuran. Hair stand samples were collected from two men approximately 1 year after alachlor intoxication for case 1, and 14 days after the last exposure for case 2. Hair analysis was performed using a liquid chromatography-tandem mass spectrometry method. In case 1, alachlor was detected in the 5 analysed hair segments (concentrations between 12 and 136 pg/mg) and its metabolites were not detected. In case 2, carbofuran and its main metabolite (3-hydroxycarbofuran) were detected in the hair strand (global analysis) at the concentrations of 207 and 164 pg/mg, respectively. However, additional data are required in order to interpret such results.     

Application of discriminant analysis to differentiate between incorporation of cocaine and its congeners into hair and contamination

The requirement to differentiate between incorporation and external contamination of drugs into hair is undisputed, in particular when dealing with compounds which are administered by sniffing or inhalation (e.g. cocaine). With the aim of making this discrimination, hair samples from cocaine (COC) users (group IN) and seized cocaine samples (group OUT) were compared regarding the parameters benzoylecgonine (BZE), ecgonine methyl ester (EME), ecgonine (ECG), anhydroecgonine methyl ester (AEME), cocaethylene (CE) and norcocaine (NCOC). Since most of these compounds may be minor by-products of COC or be formed by biotransformation or chemical degradation, the stability of each substance was carefully examined. COC was found to be converted into significant amounts of BZE, EME and ECG even under mild extraction conditions, while traces of NCOC proved to be a ubiquitous by-product of COC. Cocaine positive hairs and seized cocaine samples (diluted to relevant concentrations) were equally preprocessed and analyzed by LC-MS-MS. Out of the metabolites listed above, NCOC, CE and AEME (each normalised to COC) were significantly increased in the incorporation group (i.e. hair samples from cocaine users). Based on this approach, a statistical discriminant analysis enabled us to make a prediction (and estimation of uncertainty) for each cocaine positive hair sample as to its likelihood of belonging to the group of cocaine users or of being contaminated.     

左归饮水提物对大鼠Cyp2b1、Cyp2c11、 Cyp2c7亚酶活性的影响

目的 采用Cocktail探针药物法评价左归饮对大鼠体内细胞色素氧化酶P450亚型酶Cyp2b1、Cyp2c11和Cyp2c7代谢活性的影响,为左归饮的开发应用提供实验依据。方法 选用阿莫地喹、安非他酮、奥美拉唑分别作为Cyp2b1、Cyp2c11和Cyp2c7的探针底物,制备成Cocktail混合溶液。将SD雄性大鼠分为实验组和空白组,实验组每日清晨灌服高、中、低剂量左归饮溶液,空白组每日清晨灌服空白溶剂,均连续给药14 d。第14天清晨尾静脉注射混合探针药物后于各时间点测定大鼠体内探针药物的血药浓度。经DAS 2.0软件计算探针药物的药物代谢动力学参数,评价左归饮对大鼠体内代谢酶活性的影响。采用LC-MS/MS法同时测定3种探针药物的血药浓度。结果 与空白组比较,高、中、低剂量左归饮连续给药对Cyp2b1、Cyp2c7和Cyp2c11酶活性的影响均无统计学意义（P＞0.05）。结论 左归饮配伍其他药物使用对代谢酶无明显影响,左归饮可在临床上广泛地配伍应用。     

钩藤碱在Caco-2细胞模型中吸收规律研究

目的 建立体外模拟体内肠道细胞——人结肠癌上皮细胞（the human colon adenocarcinoma cell line，Caco-2）的Transwell模型，以此研究钩藤碱在Caco-2细胞模型上的跨膜转运特征。方法 采用聚酯碳酸酯膜连续培养Caco-2细胞21 d，形成致密的单层细胞模型。然后对影响钩藤碱在Caco-2细胞模型上转运特征的因素(包括浓度、时间及跨膜转运蛋白P-糖蛋白)进行考察；同时采用液相色谱-质谱法对溶液中钩藤碱的含量进行测定。结果 钩藤碱在Millicell系统上的转运量具有一定的浓度-时间依赖性，主要以主动转运的方式进行吸收，且随着时间和药物浓度的增加，转运量明显增加。不同浓度钩藤碱从细胞基底侧（basolateral, BL）到细胞刷状缘侧（apical，AP）的表观渗透系数（apparent permeability coefficient, Papp）与AP侧到BL侧的Papp比值均在1.5以上，且钩藤碱从BL侧到AP侧的外排速率明显高于从AP侧到BL侧的吸收速率。当加入P-糖蛋白抑制剂后，药物由BL侧向AP侧的转运量显著减少，而由AP侧到BL侧的转运量显著增加。结论 钩藤碱在Caco-2细胞上转运存在一定的浓度及时间依赖性，且P-糖蛋白介导钩藤碱在Caco-2细胞上转运。     

健胎液对模型小鼠着床期子宫内膜容受性的影响

目的:研究健胎液对胚泡着床障碍小鼠着床期子宫内膜容受性的影响。方法:于妊娠第4天9∶00利用米非司酮建立胚泡着床障碍小鼠模型,予健胎液进行干预,于妊娠第4天21∶00处死小鼠,观察小鼠胚泡着床率及平均着床胚泡数,采用放免法检测血清及子宫组织雌二醇、孕酮的含量,采用逆转录聚合酶链式反应(RT-PCR)检测子宫ERmRNA、PRmRNA表达。结果:与模型组相比,中药组着床率和平均着床胚胎数均显著升高,与对照组无显著差异;模型组子宫内膜发育不良,中药组子宫内膜的发育与正常组相似;3组血清及子宫组织的雌二醇和孕酮的含量无显著改变;中药组子宫ER mRNA、PR mRNA表达均较模型组显著提高,与正常组比较差异无显著性。结论:健胎液通过促进子宫内膜发育,调节胚泡着床障碍小鼠子宫组织ER、PR基因的表达,改善子宫内膜容受性,利于胚泡着床。     

Adaptation and evaluation of the PrepFiler™ DNA extraction technology in an automated forensic DNA analysis process with emphasis on DNA yield, inhibitor removal and contamination security

Within the initial step of the forensic DNA analysis process, the DNA extraction efficiency and especially the removal of potential PCR inhibitors is crucial for subsequent steps, e.g. quantification by real-time PCR and amplification of short tandem repeats (STRs). The protocol of the PrepFiler™ Forensic DNA Extraction Kit was optimized for the application on a Tecan liquid handling workstation Freedom EVO^® 150. This modified application of the PrepFiler™ technology was compared with respect to DNA yield, sensitivity and the ability to remove potential PCR inhibitors to an established routine method working on the same liquid handling workstation based on ChargeSwitch^® Technology (CST) from Invitrogen.     

孕康灵对动物子宫收缩活动的影响

The authors of. this article have made an observation on the effects of a compound liquid -Compound Liquid for Fetus Protection - on contraction of the nonsevered and severed animal wombs.The results show that it can block nonidiosya-cratically the contraction of the smooth muscles of the womb caused by oxytocin, acetylcholine and histamine so as to relax the excited womb muscles. Thus it can be regarded as being effective to some extent in fetus protection.     

复方槐花口服液治疗轻度混合痔临床观察

目的 观察复方槐花口服液治疗轻度混合痔的临床疗效。方法 将符合纳入标准的轻度混合痔患者150例按随机数字表法分为复方槐花口服液治疗组、痔疮片及地奥司明片对照组，每组50例，观察3组患者出血、疼痛、肛门肿物脱出评分及临床疗效。结果 与治疗前比较，3组患者治疗后出血、疼痛评分均显著降低，差异有统计学意义（P＜0.05）；复方槐花口服液组治疗前后出血、疼痛评分差值显著大于痔疮片组和地奥司明片组（P＜0.05）；3组患者治疗前后肛门肿物脱出评分比较，差异无统计学意义（P＞0.05）。3组患者临床疗效分布比较，差异具有统计学意义（P＜0.05）。结论 复方槐花口服液对轻度混合痔具有良好的临床疗效。     

Successful validation of a fully automated sample lysis workstation

The Netherlands Forensic Institute recently validated a fully automated system for sample lysis, the AutoLys workstation and AutoLys tubes, developed in collaboration with Hamilton Robotics. We have carried out the developmental validation of the AutoLys system and concluded it offers a solution to drastically reduce user-errors, increase DNA yields, gain in productivity, while maintaining quality. This article reports on the study setup and discusses the results of comparing the AutoLys workstation to the conventional (manual) method. The AutoLys system proved to successfully process all commonly found forensic sample types, producing high quality extracts, containing on average 12.1% more DNA than manually extracted samples (as determined by qPCR and STR-profiling). The optimized robotic protocol minimizes the risk on cross-contamination (no cross-contamination detected in over 500 samples tested) and retains the flexibility to adjust chemically relevant variables specific to a laboratory's individual requirements. The fully automated process enables complete sample-tracking, drastically reducing the risk on user-errors. The NFI is currently validating an on-deck magnetic bead-based purification of the lysates.     

反相高效液相色谱法测定茵栀黄口服液中栀子苷的含量

目的：测定茵栀黄口服液中栀子苷的含量。方法：采用反相高效液相色谱法，C18-ODS柱为色谱柱，甲醇-水(20：80)为流动相，流速为1．0ml／min，检测波长：230nm。结果：线性范围为0．6228～1．557μg／ml(r=0．9998)，平均回收率为100．15％，RSD为1．63％(n=5)。结论：本法操作简便，准确可靠。