Utility of immunoassay in drug screening in skeletal tissues: sampling considerations in detection of ketamine exposure in femoral bone and bone marrow following acute administration using ELISA

Detection of ketamine exposure in skeletal tissues by automated enzyme-linked immunosorbent assay (ELISA) and gas chromatography with electron capture detection (GC-ECD) is described. Rats (n = 18) received 0, 15, 30, or 75 mg/kg ketamine hydrochloride acutely (i.p.), and were euthanized within 15 min or 1 h. Ketamine was extracted from ground femoral bone by methanolic incubation followed by liquid-liquid extraction (LLE), while marrow was homogenized in alkaline solution, and then underwent LLE. Extracts were analyzed by ELISA, and subsequently by GC-ECD following derivatization with trifluoroacetic acid anhydride. The effect of tissue type (i.e., diaphyseal bone vs. epiphyseal bone vs. bone marrow) on the immunoassay response was examined through determination of binary classification test sensitivity (S) and measurement of the relative decrease in absorbance (%DA, drug-positive tissues vs. drug-free controls) in each tissue type. The %DA varied significantly between different tissues examined under a given dose condition, and generally decreased in the order marrow > epiphyseal bone > diaphyseal bone, at all dose levels examined. Measured S values for marrow, epiphyseal bone, and diaphyseal bone were 100%, 77%, and 23%, respectively (75 mg/kg dose). These results suggest that the type of skeletal tissue sampled and position sampled within a given bone (diaphyses vs. epiphyses) are important parameters in drug screening of skeletal tissues.     

中国人四肢长骨骨性标志间各段长度与总长关系的分析

利用碎裂的长骨片段 ,计算长骨的总长。选择中国人长骨 5 0 0余例 ,取其骨性标志分段测量记录 ,将测得的数据输入计算机 ,用社会科学软件包 (SPSS)中全回归法进行统计学处理。在得出的 33个多元回归方程中 ,方程1、 9、 18、 2 6证明了骨性标志间各段长度与总长的正相关关系 ,其余 2 9个方程是在有选择的略去 1个变量和因长骨近端、远端或两端缺损使 1个或 1个以上变量缺失的情况下得出。根据碎裂长骨片段骨性标志间的长度测量值 ,应用多元回归方程 ,可计算出长骨的总长     

青少年骨关节X线片的骨龄研究

通过对11～16周岁女性及13～20周岁男性青少年左侧肩、肘、腕、髋、膝、踝关节拍摄X线片,进行了青少年的骨龄研究.根据骨骼生长发育的特点,提出了各大关节X线片判定骨龄的观察指标及分级标准,并使用SPSS软件包进行数据处理,建立了青少年利用6大关节骨骺愈合程度判定女性是否已满14周岁、男性是否已满16周岁或18周岁的判别分析方程.     

针刀骨减压合消痹颗粒治疗膝关节骨性关节炎46例

目的 观察针刀骨减压联合中药内服治疗膝关节骨性关节炎的临床疗效。方法 将92例膝关节骨性关节炎患者随机分为治疗组和对照组,每组各46例,治疗组予以骨减压联合消痹颗粒内服,对照组予以关节腔注射玻璃酸钠注射液。治疗5周后,观察并比较两组的临床疗效,并采用膝关节骨性关节炎症状分级量化评分、西安大略和麦克马斯特大学骨性关节炎指数（Western Ontario and McMaster Universities osteoarthritis index,WOMAC）和简式McGill疼痛问卷（short-form McGill pain questionnaire, SF-MPQ）评价两组的疗效。结果 治疗组总体临床疗效显著优于对照组(P＜0.05),治疗组在降低膝关节骨性关节炎症状评分、WOMAC评分和SF-MPQ各维度评分方面显著优于对照组(P＜0.05,或P＜0.01)。结论 针刀骨减压联合中药内服治疗膝关节骨性关节炎疗效显著,可改善症状、减轻疼痛、提高患者生活质量。     

DNA analysis of the last Brazilian unknown soldier’s remains buried in Pistoia (Italy)

During World War II, many nations took part in the war. Among the supporters of the Alliance there was also Brazil. In August 1944, under the leadership of President Getúlio Vargas, Brazil declared war on Nazi Germany and took part in the Italian campaign by sending many troops to support the Allies in the Central Italy. Once the conflict was over, the deceased Brazilian soldiers were buried in Pistoia, a few kilometers from Florence. But only in 1960 the Brazilian government authorized the transfer of the dead soldiers to their homeland. Five years later, during the building of the Brazilian Military Votive Monument, still in the Pistoia cemetery, a last body was found but could not be identified: so he was buried as an “unknown soldier”. In December 2012, the Brazilian Embassy in Italy asked for performing forensic genetics analysis for identification purposes on the remains of this last unknown Brazilian soldier. After almost 70 years a complete short tandem repeats (STR) profile was obtained, useful for any relatives searching.Key points:

• Identification of the last Brazilian Unknown Soldier buried in Italy.
• DNA analysis on 70 years old skeletal remains.
• Brazilian soldier’s history during World War II.

     

舌骨假阳性骨折12例分析

目的 探讨舌骨假阳性骨折形成的原因以及法医学鉴定要点. 方法 整理12例在实际检案中舌骨假阳性骨折案例资料并分析其形成原因. 结果 舌骨假阳性骨折主要是由于检验时人为操作（4例）以及先天性分离型舌骨（8例）误判所致. 结论 通过骨折周缘肌肉、软组织出血以及骨折断端检验,可以对人为操作因素致舌骨骨折、分离型舌骨等舌骨假阳性骨折与真性骨折进行区分.     

Colonization of diatoms in and on porcine bone substrate and considerations of diatom ecology for forensic science

The presence of diatom algae in bone marrow has been used as forensic evidence of drowning for several decades; however, these studies are based on known or suspected recent drowning events. This study addresses the potential for diatoms to enter the bone marrow of skeletal remains, that is, de-fleshed long bones post-mortem. In laboratory and field experiments, bones were either inflicted with two access points by a cut and acid pitting or left intact. The bones were submerged in water for at least 1 week and up to 3 months. Samples of the bone surface and marrow were inspected for diatoms. The analysis considered the time required for diatoms to enter marrow and whether genus characteristics like size or mobility affect entry. The presence of an access point influenced diatom entry in that bones without an introduced access point had zero to one diatom present in the marrow, whereas a bone with an access point had >150 diatoms present in the marrow. The results of both laboratory and field phases suggest that diatoms will reliably colonize bone in as quickly as 1 week, establishing and maintaining communities for at least 3 months. However, the bone surface assemblages differ from the source community. Bone marrow displayed even more restrictive access to diatom colonization, resulting in communities dominated by small raphid diatoms. Based on these findings, we suggest some caveats on the use of diatoms as trace evidence in forensic science with recommendations for future avenues of research.     

Inter-species variation in bone mineral behavior upon heating

The characterization of inter-species variation in bone mineral (b-HAP) is of relevance to forensic science and archaeology, but has not previously been widely explored. Results of an investigation into unheated bone mineral and behavior of bone upon heating for 12 animal species (including human) demonstrate that b-HAP characteristics, quantitatively measured using X-ray diffraction (XRD) analysis, exhibit significant inter-species variation. Human bone was found to be significantly different to all other species in terms of b-HAP lattice parameter values from unheated and heated bone and in terms of recrystallization behavior of b-HAP upon heating bone to 600°C. The amounts of b-HAP thermal decomposition products were also significantly different for human bone heated to 1400°C compared to those obtained for most other species. Therefore, there is potential for the development of an XRD-based method of species identification, particularly one that distinguishes human from non-human bone.     

Forensic scatology: preliminary experimental study of the preparation and potential for identification of captive carnivore scat

Carnivore scats recovered from animal attack and/or scavenging contexts frequently contain forensic evidence such as human bone fragments. Forensic cases with carnivore involvement are increasingly prevalent, necessitating a methodology for the recovery and analysis of scat evidence. This study proposes a method for the safe preparation of carnivore scat, recovery of bone inclusions, and quantification and comparison of scat variables. Fourteen scats (lion, jaguar, lynx, wolf, and coyote) were prepared with sodium-acetate-formalin fixative; analytical variables included carnivore individual, species, body size, and taxonomic family. Scat variables, particularly bone fragment inclusions, were found to vary among carnivore individuals, families, species, and sizes. The methods in this study facilitate safe scat processing, the complete recovery of digested evidence, and the preliminary identification of involved animals. This research demonstrates that scat collected from forensic contexts can yield valuable information concerning both the victim and the carnivore involved.