仔猪睾丸组织体外培养方法的建立

以30日龄仔猪睾丸组织块为培养对象,应用飞利普TECNAI10电子显微镜对培养前后的生精细胞类别进行了观察,对培养后的组织块进行生精细胞分化程度鉴定,建立了仔猪睾丸组织生精细胞爬片体外培养模型。培养前后电镜观察及HE染色结果显示,培养前细胞种类主要有2种:支持细胞和精原细胞,有少数几个初级精母细胞;培养第20 d的睾丸组织块HE染色观察到3个精子细胞,电镜观察到2个精子细胞。证实,在没有添加睾酮的情况下,此培养体系能够提供睾丸组织块生精细胞分化所需要的条件,使非精子细胞类生精细胞减数分裂为精子细胞。     

胚兔牙囊发育生物行为的组织形态学观察

研究胚兔的牙囊发育生物行为与人的牙囊发育是否相似 ,为人口腔疾病的研究提供模拟实验动物模型 ,采用光镜、电镜及激光扫描共聚焦显微镜对 4 3只胚兔的牙囊发育过程进行形态学观察。结果显示 ,牙囊来源于外胚间质 ,通过胚胎诱导形成牙支持组织 ,分化产生成牙骨质细胞、成骨细胞、成纤维细胞 ,形成牙骨质、固有牙槽骨、牙周韧带及其基质 ,对牙齿发育阶段具有保护和稳定作用 ,调控未来形成的牙周结构 ,在早期发育阶段首先出现 ,与人牙囊发育基本相似 ;最终牙囊在牙冠的顶部形成索引带 ,诱导继承牙萌出 ,在牙冠的根部形成牙支持组织 ,一部分牙囊细胞有程序性细胞死亡 ,以维持牙囊发育期内环境的生理平衡     

Determination of MDMA, MDA, MDEA and MBDB in oral fluid using high performance liquid chromatography with native fluorescence detection

This paper describes the analytical methodology for the determination of MDMA, MDA, MDEA and MBDB in oral fluid. After a liquid–liquid extraction, the analysis was carried out by high performance liquid chromatography (HPLC), with fluorescence detection. The detector wavelength was fixed at 285 nm for excitation and 320 nm for emission. The mobile phase, a mixture of phosphate buffer (pH = 5) and acetonitrile (75:25), and the column, Kromasil 100 C8 5 μm 250 mm × 4.6 mm, allowed good separation of the compounds in an isocratic mode in only 10 min. The method was validated and showed good limits of detection (2 ng/mL) and quantitation (10 ng/mL) for all the amphetamine derivatives. No interfering substances were detected. A stability study of these compounds in oral fluid stored at three different temperatures (−18, 4 and 20 °C) over 10 weeks was conducted, showing a time-dependent degradation of the four compounds.     

Combined Collection and Analysis of Inorganic and Organic Gunshot Residues

Gunshot residue (GSR) analysis and their interpretation provide crucial information on a criminal investigation involving the use of firearms. To date, several approaches have been proposed for the implementation of a combined sampling and analysis of inorganic (IGSR) and organic GSR (OGSR). However, it is not clear at this stage if concurrent analyses of both types of residue might be detrimental to the analysis of IGSR currently applied in forensic laboratories. Thus, this work aims to compare and evaluate three different protocols for the combined collection and analysis of IGSR and OGSR. These methods, respectively, involve the use of a modified stub (with two halves, one for the detection of IGSR and the other for the analysis of OGSR); the sequential recovery of GSR with two stubs mounted with different adhesives (double-sided carbon tape and Tesa® TACK) and the sequential analysis of IGSR and OGSR from a single carbon stub following carbon deposition. The detection of IGSR was carried out using SEM-EDX, while OGSR analysis was performed using ultra-high-performance liquid chromatography–tandem mass spectrometry (UHPLC-MS/MS). Obtained results for experiments performed using Geco Sinoxid^® ammunition indicated that sequential analysis was the most suitable protocol for the combined collection and analysis of both IGSR and OGSR. A higher number of inorganic (characteristic and consistent) particles and higher concentrations of ethylcentralite, N-nitrosodiphenylamine, diphenylamine, and nitroglycerin were recovered with this method.     

紫外荧光加分色照相拍摄彩色背景上汗潜指印的探讨

在拍摄彩色背景上汗潜指印时。传统的方法是在汗潜指印上刷显有色荧光粉．再利用紫外线作为激发光源激发有色荧光粉发出荧光图像的照相方法采加以解决．这种方法是在汗液指印比较清晰的情况下可以获得一定的效果。但随着反侦查意识的增强，案犯在现场遗留的清晰痕迹物证将会越来越少．因此，提取现场疑难痕迹物证特别是在复杂背景上的汗潜指印将成为刑事技术人员不得不面临的问题。本文探讨的就是在复杂背景上的汗潜指印用传统紫外荧光照相法效果不理想的情况下．再加用分色照相的方法将二者融合在一起来解决这个问题的实验探讨。     

尿液分析仪检验血尿在法医学中应用研究

尿液分析仪检验血尿在法医学中的应用价值。应用尿液分析仪干化学法及尿沉渣显微镜法对 418例患者尿样红细胞成份同时进行检测 ,结果表明 ,尿液分析仪检验结果 :阴性 2 76例、“ +” 5 8例、“ ++” 35例、≥“ ++” 84例 ,尿沉渣镜检法相应程度的检验结果分别为阴性 2 74例、“ +-”和“ +” 31例、“ ++” 1例、≥“ ++” 37例 ,相同的程度符合率分别为 99 2 6 %、 5 3 45 %、 2 86 %、 44 0 5 %。两法阳性与阴性结果及轻伤程度 (≥ ++)与轻微伤程度 (≤ +)结果比较均有高度显著性差异 (P <0 0 1)。尿液分析仪检验阳性率高于尿沉渣镜检法 ,且表现血尿程度略严重。尿液分析仪检验结果阴性和“ +”与尿沉渣镜检法 0～ 3/HP (阴性或少许 )程度一致 ,属正常范围。目前 ,法医学鉴定中用尿液分析仪检验结果 2 5 / μl ( ++)相当于尿沉渣镜检法红细胞 10～ 30 /HP ( ++) ,引用《人体轻伤鉴定标准 (试行 )》第 36条进行法医学鉴定是不准确的 ,不能将尿液分析仪完全替代尿沉渣镜检法用于法医学鉴定     

荧光标记复合扩增毛细管电泳法在SNP分型中的应用

目的采用荧光标记复合扩增毛细管电泳法，对辽南地区汉族人群13个SNP进行等位基因频率调查，并评价其法医学应用价值。方法选择13个双等位基因SNP，应用荧光标记片段长度差异等位基因特异性复合扩增SNP分型方法，对辽南地区汉族人群进行群体调查。结果每个SNP纯合子为单一产物峰，杂合子则为长度不同的两个产物峰。不同位点扩增产物长度不同，根据产物长度和产物峰数量进行SNP分型，其结果与直接测序完全一致。同时获得辽南地区汉族人群13个SNP等位基因频率。结论采用荧光标记复合扩增毛细管电泳法进行SNP分型，方法简单实用，在法医学个人识别领域具有较高的应用。     

Chemical trace XRF analysis to detect sharp force trauma in fresh and burned bone

Forensic anthropologists may not always be able to differentiate heat-induced fractures from fractures with other aetiologies, namely sharp force traumas, with clear nefarious impact on medico-legal conclusions. The objective of this study was to experimentally investigate if blade chemical traces are transferred to defleshed bone tissue and if they remain there after a burning event. This was accomplished by prompting sharp force traumas in 20 macerated fresh pig ribs with five different instruments, namely a stainless steel knife, an artisanal knife and a ceramic kitchen knife, a small axe and a large axe. Another pig rib was used as control, not being subjected to any trauma. All instruments were probed by X-ray fluorescence (XRF) to establish the composition of each blade. Bone samples, both pre-burned and post-burned (at 500 °C, 700 °C, 900 °C and 1100 °C), were then probed by XRF. All sharp force instruments left detectable chemical traces on pre-burned bone, although not in all samples. Furthermore, traces were still detected after experimental burning in most cases. Potentially, XRF can provide relevant information about the aetiology of fractures in burned and unburned bones, although the effect of soft tissues and diagenesis must still be investigated.     

6FAM-HEX-TMR-ROX四色荧光分析体系的构建

目的为在310基因分析仪上进行6FAM-HEX-TMR-ROX四色荧光STR复合扩增分型建立基础条件。方法以pGEM载体作为靶DNA,设计引物扩增获得500~80bp的13个长度不等DNA片段,以这些片段纯化物作为模板DNA,分别扩增获得6FAM、HEX、TMR和ROX标记的M atrix标准物,用4种M atrix标准物在310基因分析仪上构建可用于6FAM-HEX-TMR-ROX四色荧光分析的M atrix文件。结果扩增所得的13个非标记片段,长度分别为80、100、120、140、160、180、200、220、260、300、340、400、500bp,在非变性聚丙烯酰胺凝胶连续电泳-银染凝胶上均表现为单条带,6FAM、HEX、TMR、ROX分别标记的片段通过310基因分析仪检测均为单峰。混合ROX标记的80、120、180、220、260、300bp的6个片段获得的内标,显示出良好的线性关系。结论建立了有效的6FAM-HEX-TMR-ROX四色荧光分析M atrix,为进行多个STR基因座荧光标记复合扩增检测奠定了基础。     

Assessment of Maceration Techniques Used to Remove Soft Tissue from Bone in Cut Mark Analysis

Maceration techniques employed in forensics must be effective without compromising the bone's integrity and morphology, and prevent destruction of evidence. Techniques must also be fast, safe, easily obtainable and inexpensive; not all techniques currently employed are appropriate for forensic use. To evaluate the most suitable approach, seven techniques including current and new methodologies were applied to fresh, fleshed porcine ribs exhibiting cut marks. A sample size of 30 specimens per technique was examined under scanning electron microscopy at the cut mark and the surrounding uncompromised regions; a scoring system of effectiveness was applied. The previously unpublished microwave method fared best for bone and cut mark preservation. Sodium hypochlorite destroyed cut marks, and was deemed unsuitable for forensic analysis. No single technique fulfilled all criteria; however, this study provides a benchmark for forensic anthropologists to select the most appropriate method for their situation, while maintaining the high standards required by forensic science.