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。     

猪ERK6基因编码区的克隆及组织表达谱分析

利用RT-PCR方法扩增了猪ERK6基因编码区的序列,将扩增产物与pMD18-T载体连接,构建了重组质粒;重组质粒经PCR、酶切鉴定后进行测序;采用Northern杂交和半定量RT-PCR方法在猪不同部位骨骼肌中分析了ERK6基因转录本的个数、大小及组织表达谱。结果显示,所克隆的猪ERK6基因片段长1 113 bp,含有1个1 104 bp的开放阅读框(ORF),该ORF编码367个氨基酸;该基因与已报道的人ERK6基因核苷酸序列相似性为90%;猪ERK6基因在肌肉组织中只有一个转录本,大小约1.5 kb。此基因在骨骼肌中的表达量最高,心、子宫次之,卵巢最低;而在脂肪、胃、肝、脾、肺、肾、十二指肠和胰腺中未见表达。结果表明,猪ERK6基因与已报道的小鼠和人ERK6基因一样,主要在骨骼中特异表达。     

兔脂肪间质干细胞的分离培养与鉴定

取兔腹股沟皮下脂肪组织,用Ⅰ型胶原酶消化,分离培养脂肪间质干细胞(MSCs),并用免疫组化和体外诱导分化方法对其表面分子标志和多向分化潜能进行鉴定。结果显示,兔脂肪组织中能够分离培养出脂肪MSCs,该类细胞表达CD29、CD44和CD105,不表达CD34、CD45及HLA-DR表面分子标志,并具有可分化为脂肪细胞、神经细胞和成骨细胞的多向分化潜能,证实兔脂肪组织中存在具有多向分化潜能的MSCs。     

Volatile Organic Compounds,Spectral Characterization and Morphology of Ammonium Nitrate Fuel Oil (ANFO) Samples

Ammonium nitrate fuel oil is an explosive mixture found in most antipersonnel landmines (APL) buried throughout the Colombian territory. During more than 50 years of internal conflict, the Colombian government has found that trained dogs are the most effective method to detect APL. However, the olfactive signature in ANFO is unknown and also if there are differences in detection related to the explosive manufacturing origin. Therefore, this work begins with the analytical validation of the method used to determine ammonia, in its derivatized form as carbamate, released by home-made ANFO using HS-SPME-GC-FID. Once validated, the method was used to identify ammonia and other organic volatile compounds present in ANFO, under laboratory and simulated field conditions. The validation process includes the evaluation of the optimum conditions for the derivation and extraction of butylcarbamate, the determination of the working ranges with linear response in FID, the limits of detection and quantification, the sensitivity, and the precision. The results of the validation established linearity and sensitivity in a concentration between 20 and 120 mg/L, as well as low limits of detection and quantification of 6.4 and 21.4 mg/L, respectively. Also, an intermediate precision of 11% for butylcarbamate with a repeatability of 8%. The validated method showed in real samples of home-made ANFO besides ammonia, the presence of low molecular methylamines, and also exhibited differences in volatile compositions according to the origin. The objective of this work is to offer a reliable analytical methodology for the extraction and analysis of volatile compounds from ANFO.     

鸡白细胞介素-2受体基因γ链的转录分析与表达

以脾单个核细胞总RNA为模板,对鸡白细胞介素-2受体基因γ链(chIL-2Rγ)进行了RT-PCR,获得了一1 047 bp的开放阅读框,编码由348个氨基酸残基组成的分子质量为37.8 ku的蛋白多肽。预测的鸡IL-2Rγ多肽链中包含4个保守半胱氨酸残基、1个WSXWS基序和7个N连接的糖基化位点。鸡IL-2Rγ与其他动物IL-2Rγ在氨基酸水平上的同源性仅为21.4%~38.2%。RT-PCR检测发现,鸡IL-2RγmRNA分布于大脑、小脑、脊髓、腔上囊、脾、胸腺、骨髓、盲肠扁桃体、腺胃、肌胃、空肠、卵黄囊憩室、回肠、盲肠、直肠、心脏、肾、肺、肝、骨骼肌和皮肤,而在十二指肠和睾丸中没有检测到其转录。构建了鸡IL-2Rγ胞外区的原核重组表达载体,进行了表达和鉴定。     

中国土家族人群头面部软组织厚度的测量

目的测量中国土家族人群头面部软组织厚度。方法156名中国湖南西部土家族成年男女(男80,女76),经拍摄头面部侧位X线片,选取正中矢状面5个部位上的10个点,测量其软组织厚度,并对所得数据进行统计学处理。结果中国土家族成年人群头面部10个点的软组织厚度测量值,男性在(3.46±0.09)mm~(9.11±0.14)mm范围,女性在(3.29±0.16)mm~(9.05±0.18)mm;除鼻下点外(男性9.06mm±0.15mm,女性9.02mm±0.19mm),男女性之间各点均有显著性差异,且下面部软组织厚度较上面部大。结论土家族成年男女性头面部软组织厚度存在着性别差异和年龄差异。     

猪和兔附红细胞体的形态学及生殖方式观察

应用光镜和电镜对猪和兔附红细胞体的显微和超微结构进行了观察。结果表明,猪和兔附红细胞体在光镜和电镜下的形态结构、大小以及在红细胞上的附着方式等均无显著差别,其形态多为球形、卵圆形,直径0.2～2.5μm,在红细胞表面以一根或数根纤丝连接成串珠状或单个寄生。在透射电镜下,首次观察到附红细胞体边缘有一段20～30nm厚的光亮区域,并且在扫描和透射电镜下同时观察到附红细胞体的出芽生殖现象。由此推测,附红细胞体的发育方式为成熟的附红细胞体以出芽的方式产生1个未成熟形态的附红细胞体,随着未成熟形态附红细胞体的发育,逐渐从成熟附红细胞体体内脱离出来,最后以纤丝与成熟附红细胞体相连接;脱离出来的未成熟形态的附红细胞体黏附到同一红细胞或临近红细胞的膜上,最终发育成1个成熟的附红细胞体。     

东方蝾螈的形态观察和解剖学研究

采用解剖学方法对8尾东方蝾螈的外形和内脏进行了系统研究。结果表明,东方蝾螈的口裂很宽,口咽腔底壁的前方有不发达的舌。食管短,黏膜上有纵行皱襞;胃呈纺锤形的长囊,肌层发达,贲门口较大;直肠粗大;肝较为发达,有胆囊;胰独立,不与肝混合,但胰管与胆管合并。肺为2个锥形的薄壁长囊,构造简单,囊壁形成蜂窝状。输尿管不直接通膀胱,尿液先到泄殖腔,然后再到膀胱。雄性大鲵无专门的输精管,而是借助于输尿管输精。雌性大鲵有卵巢1对,呈带形的囊状卵巢。     

Comparison of the Columnar‐Thin‐Film and Vacuum‐Metal‐Deposition Techniques to Develop Sebaceous Fingermarks on Nonporous Substrates,

Both the columnar‐thin‐film (CTF) and the vacuum‐metal‐deposition (VMD) techniques for visualizing sebaceous fingermarks require the deposition of a material thereon in a vacuum chamber. Despite that similarity, there are many differences between the two techniques. The film deposited with the CTF technique has a columnar morphology, but the film deposited with the VMD technique comprises discrete islands. A split‐print methodology on a variety of fingermarked substrates was used to determine that the CTF technique is superior for developing fingermarks on clear sandwich bags and partial bloody fingermarks on stainless steel. Both techniques are similar in their ability to develop fingermarks on glass but the CTF technique yields higher contrast. The VMD technique is superior for developing fingermarks on white grocery bags and the smooth side of Gloss Finish Scotch Multitask^? tape. Neither technique worked well for fingermarks on black garbage bags.     

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.