The Stability of 4-Chloromethcathinone in Blood and Vitreous Humor

We present results of our study on the stability of 4-chloromethcathinone (4-CMC) in authentic postmortem peripheral blood and vitreous humor samples. The stability of 4-CMC was determined in postmortem blood samples (for a period of 90 days) and vitreous humor (30 days) at three different temperatures: −15°C, +4°C, and + 23°C. The analyses were carried out using ultra-high-performance liquid chromatography coupled with triple-quadrupole tandem mass spectrometry (UHPLC-QqQ-MS/MS). In both materials, the lowest 4-CMC stability was demonstrated at room temperature. The blood samples stored in a freezer (−15°C) showed stability for the entire study period (90 days), while in the case of the vitreous humor sample stored at the same temperature the concentration of the substance decreased by 53% after 30 days. The study carried out in authentic postmortem blood and vitreous humor samples confirms the previous reports of 4-CMC instability in biological material. Authors suggest that the biological material should be stored frozen until analyses are carried out as soon as possible after collection of the material.     

A novel method for cyanide quantification in human whole blood using ion chromatography with amperometric detection and its application to cyanide intoxication cases

Cyanide is a highly toxic agent that has been frequently used for suicide in South Korea. It is also used in various industrial fields, such as metal plating, in which many accidental cyanide intoxications have occurred. To overcome the disadvantages of conventional cyanide analysis methods, a simple and fast method for the analysis of cyanide in whole blood using ion chromatography (IC) with amperometric detection was developed in this study. Whole blood samples were deproteinized, diluted, and analyzed using an IC–amperometric detection system. The limits of detection and quantitation were 0.1 and 0.2 mg/L, respectively. The method showed good linearity in the range of 0.2 to 50 mg/L with R² > 0.99. The intra- and inter-assay precision and accuracy values were <10%. The established method was successfully applied to analyze whole blood samples from three cyanide intoxication cases.     

Internal validation of reduced PCR reaction volume of the Qiagen Investigator® Argus X-12 QS Kit from blood samples on FTA cards

The onus of proof in criminal cases is beyond any reasonable doubt, and the issue on the lack of complete internal validation data can be manipulated when it comes to justifying the validity and reliability of the X-chromosomal short tandem repeats analysis for court representation. Therefore, this research evaluated the efficiency of the optimized 60% reduced volumes for polymerase chain reaction (PCR) amplification using the Qiagen Investigator® Argus X-12 QS Kit, as well as the capillary electrophoresis (CE) sample preparation for blood samples on Flinder's Technology Associates (FTA) cards. Good-quality DNA profile (3000–12,000 RFU) from the purified blood sample on FTA card (1.2 mm) were obtained using the optimized PCR (10.0 μL of PCR reaction volume and 21 cycles) and CE (9.0 μL Hi-Di™ Formamide and 0.3 μL DNA Size Standard 550 [BTO] and 27 s injection time) conditions. The analytical and stochastic thresholds were 100 and 200 RFU, respectively. Hence, the internal validation data supported the use of the optimized 60% reduced PCR amplification reaction volume of the Qiagen Investigator® Argus X-12 QS Kit as well as the CE sample preparation for producing reliable DNA profiles that comply with the quality assurance standards for forensic DNA testing laboratories, while optimizing the analytical cost.     

The impact of fentanyl on DUIDs and traffic fatalities: Blood and oral fluid data

Fentanyl has emerged as the most prolific drug in the ongoing opioid epidemic and has greatly impacted traffic safety in recent years. This study aimed to evaluate fentanyl prevalence and concentrations in blood and oral fluid in driving under the influence of drugs (DUID) cases in three different regions (i.e., Alabama, Orange County, CA, and Houston, TX) from 2017 to 2022. Furthermore, traffic fatalities were evaluated for Alabama and Orange County, CA. Fentanyl positivity in DUID and traffic fatalities increased for most years in this study. In Alabama, the prevalence of fentanyl DUID cases increased 4-fold in 2022 compared to 2017. Orange County's increase from 2017 to 2022 was 14-fold. In Houston, the increase was approximately 2-fold from 2019 to 2022. The greatest increase for all laboratories coincided with the start of the COVID-19 pandemic. In 2022, the median fentanyl DUID blood concentrations were 4.7, 11, and 4.7 ng/mL in Alabama, Orange County, and Houston, respectively. Most fentanyl cases were polydrug cases (≥90%). Methamphetamine, THC, and alprazolam were the most frequently detected drugs in combination with fentanyl. Alabama has collected oral fluid and blood in DUID cases since 2018. The detection of fentanyl in oral fluid was comparable to blood. However, 59% and 8.7% of fentanyl-positive cases had concentrations of >20 ng/mL in oral fluid and blood, respectively. Therefore, oral fluid as an alternative or supplemental specimen to blood is an attractive approach for fentanyl in DUID cases. This study contributes to understanding recent fentanyl trends and their impact on traffic safety.     

基于miRNA相对表达量自动判别月经血和外周血

目的探索有效区分月经血和外周血的miRNA最优标记组合及最佳分类模型,并构建简便快速的自动化判别软件。方法对10种miRNA(miR-451a、miR-205-5p、miR-203a-3p、miR-214-3p、miR-144-3p、miR144-5p、miR-654-5p、miR-888-5p、miR-891a-5p、miR-124-3p)在200余份月经血和外周血样本中的相对表达量以实时荧光定量PCR检测,并以7种算法模型(核密度估计、K-最近邻、逻辑回归、线性判别分析、支持向量机、神经网络、随机森林)进行数据分析,选出鉴别效果最好的标记组合及算法模型,进而构建自动判别软件。结果月经血和外周血中差别最大的三种miRNA为miR-205-5p、miR-203a-3p和miR-214-3p,使用miR144-5p与上述miRNA中的一种或两种组合可达较好区分效果,其中基于miR-144-5p、miR-203a-3p和miR205-5p所形成的“最优特征项组合一”稳健性最强。7种算法模型中最佳分类模型为核密度估计模型,其次为逻辑回归模型。结论本研究建立的自动判别软件界面友好、使用简单,适合辅助法医检验关于月经血和/或外周血判别分析的计算,便利于法医物证工作,有较大的推广应用价值。     

死后血浆和溶血样本中IgE的稳定性

目的探究死后血浆和溶血样本中免疫球蛋白E(immunoglobulin E,IgE)经过不同保存条件及冻融处理后的稳定性。方法选取39例死后48h内非冷冻尸体的心血样本,其中20例取血浆样本,19例取全血制成溶血样本。将样本置于-20℃、4℃、25℃条件下保存28d及-80℃条件下保存1年以探究IgE在不同保存条件下的稳定性。利用5次反复冻融处理样本探究死后血浆及溶血样本IgE的冻融稳定性。应用电化学发光法检测处理前后血浆和溶血样本中IgE的浓度。结果血浆样本中IgE在-20℃、4℃、25℃3种保存条件下的降解率接近,28d后降解率均值在15%左右,相同条件下溶血样本中IgE降解速度快于血浆(P<0.05),并且在25℃条件下降解率高于另外两种(P<0.05)。血浆样本于-80℃冷冻保存1年后的浓度与冷冻前相比差异无统计学意义(P>0.05),而溶血样本在-80℃冷冻保存1年后浓度有所降低(P<0.05)。死后血浆和溶血样本在反复冻融5次后检测结果与冻融前相比差异无统计学意义(P>0.05)。结论死后血浆及溶血样本中IgE具有良好的冻融稳定性。IgE在死后血浆样本中的稳定性优于溶血样本,因此在实际案例中建议及早分离血浆保存待测。     

UPLC-MS/MS法同时检测全血中16种除草剂

目的建立全血中16种除草剂的超高效液相色谱-串联质谱(UPLC-MS/MS)同时检测方法,为除草剂中毒案事件及其他刑事案件血液中该16种除草剂的检验鉴定提供依据。方法取200μL的血液,加入800μL乙腈-水(体积比80/20),进行蛋白沉淀后,采用Acquity BEH C18(2.1mm×100mm,1.7μm)色谱柱,以水(5mmol/L的甲酸铵,0.1%的甲酸)-乙腈为流动相进行梯度洗脱,采用电喷雾离子源(ESI)、多反应监测(MRM)正离子模式对16种化合物进行检测。结果在1~200ng/mL范围内线性关系良好,R2均大于0.996;基质效应(ME%)为85.2%~104.4%,相对标准偏差(RSD%)为0.72%~4.84%;仪器检出限(IDLs)为0.2~2 ng/mL(信噪比S/N≥3),方法检出限(MDLs)为0.5~3ng/mL(信噪比S/N≥3),最低定量限(LOQs)为1~7ng/mL(信噪比S/N≥10)。结论本实验建立的全血中16种除草剂同时检验方法,前处理简便快捷、回收率高、精密度好、方法检出限低,可作为该16种除草剂中(投)毒案件的检验方法。     

灸刺督脉治疗气虚血瘀型缺血性脑卒中临床观察

目的 观察评价灸刺督脉疗法治疗气虚血瘀型缺血性脑卒中的临床疗效。方法 将气虚血瘀型缺血性脑卒中患者41例，随机分为对照组（20例）和治疗组（21例）。两组均予以基础病管理，对照组采用脑卒中气虚血瘀型常规针刺治疗，取穴为顶颞前后斜线、极泉、尺泽、内关、委中、足三里、三阴交、气海、血海；治疗组在常规针刺治疗基础上加用灸刺督脉疗法，取通督七穴（水沟、神庭、百会、风府、大椎、命门、腰阳关），其中百会艾灸，大椎刺络，其余腧穴予以针刺治疗。治疗周期为14 d。疗程结束后以美国国立卫生研究院卒中量表（National Institute of Health stoke scale，NIHSS）及改良Rankin量表（modified Rankin scale，MRS）评分结果评价两组的临床疗效。结果 两组患者治疗后NIHSS及MRS评分均较治疗前显著降低（P<0.05），治疗组NIHSS及MRS评分差值与对照组比较，差异均有统计学意义（P<0.05）。两组临床疗效分布比较，差异有统计学意义（P<0.05）。结论 灸刺督脉疗法治疗气虚血瘀型缺血性脑卒中临床疗效确切，优于常规针刺治疗。     

基于紫外可见积分球反射光谱法鉴定血痕经历时间

目的建立一种快速鉴定血痕陈旧度的方法。方法在实验室条件(20℃、25℃、30℃)下,使用UV-2450型紫外可见分光光度计,利用反射附件ISR-240A积分球,以BaSO_4标准白板做参比,间隔一定时间测定健康人耳静脉血血痕纱布的反射光谱,采集特定波长反射率R_(541)、R_(577),计算R_(541)/R_(577)值,使用SPSS 17.0软件进行直线拟合与回归分析。结果一定条件下8 h内,血痕经历时间与其特定波长紫外可见反射率比值回归分析的R~2均大于0.8,回归方程成立,血痕经历时间与R_(541)/R_(577)值具有高度相关性。结论所建立的方法简便、快速、无损,可靠性好,可以对实验室条件下8 h内血痕经历时间进行鉴别。     

金龙蛤蚧平喘胶囊治疗稳定期慢性阻塞性肺疾病临床观察

目的 观察金龙蛤蚧平喘胶囊治疗稳定期慢性阻塞性肺疾病（chronic obstructive pulmonary disease，COPD）的疗效。方法 将58例COPD患者随机分为治疗组和对照组，每组28例。在常规治疗基础上，分别给予金龙蛤蚧平喘胶囊与安慰剂。治疗前、治疗6个月后和治疗12个月后，分别观察两组患者的肺功能、骨密度（bone mineral density，BMD）、BODE指数、COPD评估测试（COPD assessment test，CAT）问卷评分及1年内急性加重次数。结果 治疗6个月后及治疗12个月后，两组肺功能各项参数比较，差异均无统计学意义（P＞0.05）。 治疗12个月后，治疗组股骨颈BMD较治疗前显著升高（P＜0.05），而对照组股骨颈BMD较治疗前显著下降（P＜0.05）。治疗6个月后和治疗12个月后，两组BODE指数及其因子6 min步行距离积分比较，差异均有统计学意义（P＜0.05）。两组1年内急性加重次数相比，差异具有统计学意义（P＜0.05）。结论 金龙蛤蚧平喘胶囊可有效改善稳定期COPD患者的肺功能、BODE指数及生活质量，减少患者1年内急性加重次数。