Quantitative assessment of the percent fat in domestic animal bone marrow

Measurement of the amount of fat in femoral bone marrow can provide a quantitative assessment of the nutritional status of an individual animal. An analytical method is presented for quantitating the percent fat in bone marrow from three domestic species: bovine, canine, and equine. In this procedure, fat is extracted from bone marrow using pentane, and the percent fat recovered is determined gravimetrically. Based on analyses from adult animals (normal body condition scores), the average percentage of fat in the bone marrow was >80%. In cases in which animals have been diagnosed as emaciated or exhibit serous atrophy of fat (body scores of 1 or 2), the femoral bone marrow fat was less than 20%. In domestic animals, bone marrow fat analysis can be a useful, quantitative measure that, when used in conjunction with all other data available, can support a diagnosis of starvation or malnutrition.     

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.     

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.     

硝基苯致小鼠骨髓细胞的突变作用

为探讨硝基苯对哺乳动物的致突变性,将12周龄雄性昆明小鼠随机分为染毒组(26、52、104mg/kg)、空白对照组(生理盐水)、溶剂对照组(花生油)和阳性对照组(环磷酰胺),连续灌胃染毒7 d,检测骨髓细胞微核率、骨髓细胞染色体畸变率以及外周血网织红细胞数。结果显示,随着染毒剂量的增加,骨髓细胞微核率、染色体畸变率均有增加,外周血网织红细胞数减少,各组与对照组差异均极显著(P<0.01),并呈剂量-效应关系。表明,硝基苯可导致小鼠骨髓细胞发生突变。     

Differentiation of Antemortem and Postmortem Appendicular Fractures Using Magnetic Resonance Imaging Signal Intensity Changes in Bone and Soft Tissues,

Intraosseous T1-weighted (T1W) and short-tau inversion recovery (STIR) magnetic resonance imaging (MRI) signal intensity changes—so-called bone marrow edema—may be able to differentiate antemortem and postmortem fractures in human forensic imaging. The primary objective of this study was to investigate this hypothesis using an animal model. Three juvenile Landrace pigs were anesthetized and underwent MRI of both tibiae and both radii using a 1.5 T magnet. T1W, T2-weighted (T2W), STIR, and T2* sequences were included. Antemortem fractures were induced in both tibiae and postmortem fractures in both radii, and MRI was repeated. Two board-certified radiologists blinded to fracture group jointly evaluated the images for intraosseous and soft tissue signal intensity changes. Sensitivity (Se) and specificity (Sp) in identifying antemortem fractures were calculated based on intraosseous, soft tissue, and combined intraosseous and soft tissue signal intensity changes. Intraosseous and soft tissue signal intensity changes, when present, were hyperintense in all sequences. Intraosseous hyperintensity in T1W and T2W sequences yielded Sp of 100% for antemortem fractures. Regardless of sequence, soft tissue hyperintensity was comparatively more sensitive than intraosseous hyperintensity. Sensitivity for each sequence could be maximized by assessment of soft tissue and intraosseous hyperintense signals together; for the T1W sequence, such assessment optimized diagnostic utility yielding a Se of 100% and Sp of 83%. In summary, MRI—particularly the T1W sequence—can differentiate antemortem and postmortem fractures and may be a useful adjunct to the forensic analyses of fractures.     

补肾填精法对兔骨折后不同时相骨髓间充质干细胞成骨分化的影响

目的 观察补肾填精中药对兔骨折后不同时点骨髓间充质干细胞(bone marrow mesenchymal stem cells，BMSCs)分化为成骨细胞的影响。方法 将20只白兔随机分成4组，即正常对照组、骨折模型组、中药治疗组、西药治疗组。分离培养骨折端BMSCs，取P3代BMSCs并向成骨细胞诱导分化，分别采用碱性磷酸酶（alkaline phosphatase, ALP）试剂盒、骨钙素（osteocalcin, OCN）试剂盒检测ALP活性、OCN水平。结果 随着用药时间的延长，各骨折组ALP活性和OCN水平均逐渐增强。在骨折后第14、21天，中药治疗组ALP活性、OCN水平均高于其他3组。结论 补肾填精法可促进兔骨折后不同时点BMSCs向成骨细胞分化。     

六味地黄汤通过树突状细胞Notch信号通路调节T细胞分化干预自身免疫性脑脊髓炎

目的 通过观察树突状细胞(dendritic cells,DCs)Notch信号通路对CD4+T细胞分化的影响,探究六味地黄汤干预实验性自身免疫性脑脊髓炎(experimental autoimmune encephalomyeli-tis,EAE)的机制.方法 采用主动免疫法诱导2D2小鼠EAE模型,采用磁珠分选收集...     

生血方拮抗奥沙利铂联合替吉奥方案致晚期胃癌骨髓抑制的临床观察

目的 观察自拟生血方治疗脾肾两虚型晚期胃癌行奥沙利铂联合替吉奥方案化学治疗致骨髓抑制的疗效和安全性。方法 将60例中医辨证为脾肾两虚型晚期胃癌患者随机分成对照组和观察组,每组30例。对照组予以奥沙利铂联合替吉奥方案化学治疗,观察组在对照组基础上加用自拟生血方汤剂口服及粉剂加工后外敷穴位联合治疗。观察两组患者首次骨髓抑制发生时间、骨髓抑制持续时间、0~3周骨髓抑制等级变化,采用卡氏功能状态量表（Karnofskys performance scale,KPS）、中医证候积分评价患者生活质量,观察两组患者的无进展生存期（progression- free survival, PFS）。结果 与对照组比较,观察组患者首次出现白细胞、血红蛋白、血小板下降的时间明显延长（P＜0.05）,白细胞、血红蛋白、血小板抑制持续时间明显缩短（P＜0.05）。两组患者不同时点白细胞、血红蛋白、血小板抑制等级比较,差异有统计学意义（P＜0.05）,两组患者化学治疗第21天与化学治疗第1天白细胞抑制等级变化比较,差异有统计学意义（P＜0.05）。与治疗前比较,对照组患者治疗后KPS评分显著降低（P＜0.05）。两组患者治疗前后KPS评分差值比较,差异有统计学意义（P＜0.05）。与治疗前比较,观察组患者中医证候积分显著下降（P＜0.05）,观察组中医证候积分下降程度明显大于对照组（P＜0.05）。观察组患者PFS为10个月,较对照组PFS（6个月）明显延长。结论 自拟生血方治疗晚期脾肾两虚型胃癌奥沙利铂联合替吉奥方案化学治疗所致骨髓抑制,能延迟首次骨髓抑制发生时间并缩短持续时间,降低白细胞骨髓抑制等级,在一定程度上改善晚期胃癌患者临床症状及生活质量。