羊源巴贝斯虫未定种甘肃敦煌株的发现

采用动物接种法将采自甘肃敦煌的15只绵羊血液混合后感染1只除脾绵羊,感染后逐日做血涂片检查。结果显示,在感染后第10天的血片上出现了1种大型巴贝斯虫。虫体形态以双梨子形、单梨子形、圆形及卵圆形为主。在感染后第12天,感染羊体温升至41.1℃。染虫率达到0.125%,之后逐渐下降。无菌采集该羊血液,提取全血基因组,用梨形虫通用引物进行PCR扩增全长18SrRNA基因,得到长约1 700bp的片段,将其克隆并测序(登录号:HQ730762),与GenBank中的其他巴贝斯虫的序列进行比较,发现其与新疆巴贝斯虫未定种的同源性最高,达到99.8%。本研究发现甘肃敦煌又是一个新疆羊巴贝斯虫未定种的疫源地。     

牛瑟氏泰勒虫二温式PCR检测方法的建立及应用

为建立一种能快速、特异、敏感地检测牛瑟氏泰勒虫的方法,根据GenBank中的牛瑟氏泰勒虫MPSP基因(GQ180198.1)设计合成了1对特异性引物,建立了检测牛瑟氏泰勒虫的二温式PCR方法。该方法与犬新孢子虫、猪支原体和弓形虫RH株均无交叉反应,能检测到的最低DNA含量为1.7pg/μL。结果表明,该二温式PCR检测方法具有较高的特异性和敏感性。本研究为牛瑟氏泰勒虫的快速检测提供了一种方法。     

Identification of the source of ivory idol by DNA analysis

In this study, we describe a forensic case dealing with the identification of the source of the processed ivory object by DNA analysis. Two pieces of Lord Krishna's idols from a shop were confiscated by an investigating agency of the Indian government and forwarded to us to identify the source of its origin. We succeeded in isolating DNA from both processed ivory idols by using the phenol/chloroform DNA extraction method. The extracted DNA was subjected to PCR amplification using an elephant-specific mitochondrial DNA (mtDNA) D-loop marker. DNA sequence analysis of the amplified fragment of mtDNA D-loop region confirmed that the idols were consistent with Asian elephant with 99% similarity.     

Rapid direct PCR for ABO blood typing

Many different molecular typing methods have been reported to complement routine serological ABO blood typing in forensics. However, these ABO genotyping methods are often time-consuming and call for an initial DNA isolation step that requires the use of expensive kits or reagents. We report here a rapid direct ABO genotyping method that eliminates the need for DNA extraction from fresh blood, hair, and body fluid stains before PCR. Using a fast PCR instrument and an optimized polymerase, the genotyping method-which employs a multiplex allele-specific primer set for the simultaneous detection of three single-nucleotide polymorphism (SNP) sites (nucleotides 261, 526, and 803)-identifies A, B, O01/O02, O03, and cis-AB01 alleles in around 70 min from sample collection to electropherogram. Not only will this ABO genotyping method be efficiently used in forensic practice for rapid screening of samples before full-blown multilocus short tandem repeat profiling, but it will also demonstrate an example of rapid direct genotyping of SNPs that offers the advantages of time- and cost-efficiency, convenience, and reduced contamination during DNA analysis.     

妊娠绵羊子宫内膜组织内源性反转录病毒及其受体HYAL2基因表达水平的检测

为了探究内源性反转录病毒(enJSRV)及其受体HYAL2在妊娠蒙古绵羊子宫内膜形成和发育过程中的作用,运用TaqMan实时荧光定量PCR和组织原位杂交技术对其在不同妊娠时期(30、50、70、90、110和130d)蒙古绵羊子宫内膜中的表达规律和分布定位进行了研究。实时荧光定量PCR结果显示,enJS-RV及其受体HYAL2在蒙古绵羊子宫内膜的不同妊娠时期均有表达,通过SPSS统计学软件分析可以看出,妊娠蒙古绵羊子宫内膜组织中enJSRV mRNA的表达于妊娠第30和50天相对较高,70～130d均低于对照组(30d),且差异都极显著,enJSRV与其受体HYAL2mRNA之间亦无线性相关性。原位杂交结果显示,enJSRV及其受体HYAL2mRNA在妊娠第30、50和130天蒙古绵羊子宫内膜的腔上皮细胞、腺上皮细胞、血管内皮细胞、间质及滋养层巨型双核细胞中均有阳性信号表达。表明enJSRV及其受体HYAL2表达于上皮细胞及上皮细胞来源的滋养层巨型双核细胞中,揭示enJSRV及其受体HYAL2在子宫形成过程中和系统防御中可能发挥重要作用。     

Cathepsin-L在缺血心肌中的表达

目的检测cathepsin-L(CTSL)在大鼠早期缺血心肌中的表达变化,并探讨其法医学意义。方法建立大鼠早期心肌缺血模型,设置早期缺血组(early ischemic myocardium,EIM)、非缺血组(non-ischemic myocardium,NIM)、假手术组以及空白对照组,采用real-time PCR方法检测大鼠缺血后15min,30min,1h和2h CTSL mRNA的表达量,并进行组内和组间比较。收集心源性猝死者缺血心肌和机械性损伤致死者正常心肌样本,采用免疫组化染色方法观察心肌组织中CTSL蛋白表达。结果 EIM组CTSL mRNA表达量与NIM组、假手术组及空白组相比,在15min时无统计学意义(P>0.05),在30min、1h和2h时分别升高1.4、3.1和4.5倍(P<0.05);其余3组间差异无统计学意义(P>0.05)。观察10例冠脉狭窄程度Ⅲ～Ⅳ级的心源性猝死(SCD)者缺血心肌,CTSL表达增强。结论大鼠心肌缺血后早期即可检测到CTSL mRNA的升高,并在SCD冠脉狭窄死者缺血心肌中高表达,提示CTSL可作为心肌缺血与SCD的参考指标。     

检测浮游生物叶绿素相关基因诊断溺死的实验研究

目的评估浮游生物叶绿素相关基因检测用于溺死诊断的价值。方法将18只大白兔随机分为溺死组（n=10）、死后抛尸组（n=6）和对照组（n=2）,各组分别取心血及肺、肝、肾、脑组织,分离浮游生物并提取其DNA,用PCR技术检测叶绿素相关基因EG（EG1和EG2）及SK（SK1和SK2）。同时用硝酸消化法检验肺和肝组织中的硅藻。结果溺死组心血及肺、肝、肾、脑组织中EG1分别检出9、10、9、7和8例阳性,EG2分别检出8、10、7、5和7例阳性;死后抛尸组仅在心血和肺组织中各检出1例EG1阳性;对照组各组织均未检出EG1和EG2。SK1、SK2除在溺死组心血、肺和肾有少数检出外（≤2例）,在其他组未检出扩增产物。硝酸消化法从溺死组肺、肝组织中分别有9例及3例检出硅藻,死后抛尸组仅在1例肺组织中检出。结论浮游生物叶绿素相关基因EG用于溺死诊断的阳性检出率要高于硝酸消化法,在溺死诊断中具有较高的应用价值。     

Rural development through the construction of new,nested, markets: comparative perspectives from China,Brazil and the European Union

This article discusses and compares the rural development processes and practices currently occurring in China, Brazil and the European Union. Although these are strongly rooted in the specificities of time and space, they also share important commonalities. We argue that rural development can be viewed as an evolving set of responses to market failures. A key element of these responses is that they are unfolding through the construction of new markets: a seemingly contradictory phenomenon that has, as yet, hardly been scrutinized or theoretically elaborated. We describe these newly emerging markets as ‘nested markets’ and support our argument with a careful reconsideration of the dynamics of long-established nested markets. We then extend this analysis, firstly, by arguing that the construction of such new markets occurs through a process of social struggle and, secondly, by exploring the strength of these newly emerging constellations in relation to the hegemony exerted by food empires. Our analysis puts common-pool resources, which underlie these new, nested markets, centre stage.     

A validation study of mRNA markers for skin cell identification

The objectives of this study were to develop and optimize a multiplex of three skin specific gene markers; loricrin (LOR), corneodesmosin (CDSN) and keratin 9 (KRT9) and 1 house-keeping marker, β-actin (ACTB) using an endpoint PCR assay to analyze expression data from a range of relevant samples. Marker specificity and suitability were evaluated for their inclusion in future forensic casework. The presence of the three skin mRNA markers was successfully confirmed from swabs of human skin obtained from 20 individuals at each of 6 different body sites (forehead, neck, arm, palm, leg and sole). Significant variation was observed in the relative expression of the three genes across the body sites, with some individuals consistently failing to express one or more of the targets. Inter-individual variation was also evident. Accordingly, these markers must be used with caution in the identification of skin in forensic samples.     

p33.4位点扩增片段长度多态性及其法医学应用的研究

以聚合酶链反应（PCR）、聚丙烯酰胺凝胶垂直电泳和银染法对中国100名无关个体小卫星区域p33．4位点的扩增片段长度多态性（Amp－FLPs）进行了研究，检出了8个等位基因。通过BIOTRAC系统进行数据处理．各等位基因重复单位的数目分别为7、10到15，其中在13～14之间发现一差值不足一个重复单位长度的罕见等位基因。片段长度分布于603～1115bP之间，基因频率分布于0.5～33．5％间，杂合度为64％，DP值为84．5％。对5个家庭25名相关个体进行分析，符合孟德尔遗传定律；对同一个体不同组织的DNA进行P33.4位点的分型研究表明，该技术适宜于法医物证检验。此外，本研究以Chelex处理不同检材制备DNA模板用于扩增，率先建立了比常规方法更快、更简便、更为实用的检验方法。