口蹄疫病毒A/WH/09株结构蛋白P1基因的克隆及其B细胞抗原表位的预测

对口蹄疫病毒(FMDV)A/WH/09株P1结构蛋白基因进行了扩增、克隆及序列测定。结果显示,FMDV A/WH/09株P1基因长2 208bp,包含完整的开放阅读框,编码736个氨基酸,其中VP1长636bp,编码212个氨基酸;VP2长654bp,编码218个氨基酸;VP3长663bp,编码221个氨基酸;VP4长255bp,编码85个氨基酸。采用DNAStar Protean软件对P1蛋白的二级结构、可塑性、亲水性、表面可及性及抗原指数等参数进行了综合分析,并预测其B细胞的抗原表位分布。结果表明,VP1、VP2和VP3上均有多个区域为B细胞抗原优势表位,VP4上也有少量潜在的B细胞抗原表位。与已鉴定的B细胞抗原表位相比,本试验所用方法预测的结果有较高的准确度,是一种较为先进的表位研究方法。     

金黄色葡萄球菌黏附素FnBPB-ClFA共表达质粒的构建与原核表达

采用PCR方法对编码金黄色葡萄球菌纤黏蛋白B的D区和纤黏蛋白原凝集素A的A区基因片段进行了特异性扩增,并通过重叠延伸PCR扩增了FnBPB-ClFA串联基因,构建了克隆质粒pMD19-FnB-PB-ClFA。将该基因片段定向插入到原核表达载体pET-32a(+)中,构建了表达质粒pET-FnBPB-ClFA,并将其转入宿主菌BL21(DE3)。SDS-PAGE分析表明,在1mmol/L IPTG诱导下,在51ku处出现了与目的蛋白一致的外源蛋白带。Western-blot分析表明,所表达的蛋白与目的蛋白一致。上述结果表明,该融合基因在原核细胞中成功表达。     

Sudden Death Secondary to an Undiagnosed B‐Cell Lymphoma of the Hypopharynx and Infiltration of the Inferior Constrictor Muscle

The aim of this presentation was to share an uncommon form of sudden death, suffered by a 64‐year‐old woman, due to a mechanical obstruction of hypopharynx by an undiagnosed B‐cell lymphoma, infiltrating the inferior pharyngeal constrictor muscle. A forensic approach by means of scene investigation, circumstantial data collection, autopsy, and histological and toxicological investigations led to conclude that the cause of death was asphyxia, correlated with B‐cell lymphoma of the hypopharynx. The autopsy examination highlighted the presence of a wall thickening, infiltrating, and projecting into the hypopharynx lumen. The histological analysis showed the essential finding of a B‐cell lymphoma of the hypopharynx, diffusely infiltrating the inferior pharyngeal constrictor muscle. To conclude, this case demonstrates once more that in the absence of specific data, a thorough forensic investigation including autopsy, histological examination, and circumstantial data collection is mandatory to reach a correct cause of death.     

The Upper Silesia (Poland) population and forensic usefulness of 11 autosomal STR loci

Allele frequencies, forensic parameters for the 11 STR loci in GenePrint STR Systems—Silver Stain Detection (Promega): CSF1PO, TPOX, TH01, D16S539, D7S820, D13S317, F13A01, FESFPS, VWA, F13B, LPL were determined in a sample of 529–1436 unrelated, dead and alive adults from the Upper Silesia region (Poland). Furthermore, gender identification was carried out. The values of heterozygosity (Ht), polymorphic information content (PIC), power of discrimination (PD), matching probability (PM), mean exclusion chance (MEC) and mean exclusion probability (MEP) were calculated using Dudek's computer programme (FatRec, High Technical School of Cz?stochowa, Poland). A possible divergence from HWE was determined by χ² and exact tests using Miller's computer programme (TFPGA, Northern Arizona University, Flagstaff, USA). Comparison of allele frequencies between the Upper Silesia population and other Polish populations was performed with Carmody's test.     

Developmental validation of a Cannabis sativa STR multiplex system for forensic analysis

A developmental validation study based on recommendations of the Scientific Working Group on DNA Analysis Methods (SWGDAM) was conducted on a multiplex system of 10 Cannabis sativa short tandem repeat loci. Amplification of the loci in four multiplex reactions was tested across DNA from dried root, stem, and leaf sources, and DNA from fresh, frozen, and dried leaf tissue with a template DNA range of 10.0-0.01 ng. The loci were amplified and scored consistently for all DNA sources when DNA template was in the range of 10.0-1.0 ng. Some allelic dropout and PCR failure occurred in reactions with lower template DNA amounts. Overall, amplification was best using 10.0 ng of template DNA from dried leaf tissue indicating that this is the optimal source material. Cross species amplification was observed in Humulus lupulus for three loci but there was no allelic overlap. This is the first study following SWGDAM validation guidelines to validate short tandem repeat markers for forensic use in plants.     

C/S、B/S结构相结合的公安院校远程招生系统设计

本文提出了建立基于C/S、B/S体系结构相结合的公安院校远程招生系统,给出了总体设计方案,包括系统结构、系统模块功能和数据库表结构,提出了系统设计的原则,介绍了系统所采用的软件平台和所使用的基本技术。     

提单与票据之比较研究

有关提单的法律规定尚不完整 ,因提单和票据实质上有许多相似之处 ,人们可以利用票据法的一些规定来解决提单的一些问题。本文比较研究了提单和票据的法律性质以及二者在转让中的前后手权利优劣等事项。     

凉血解毒汤联合窄谱中波紫外线照射治疗玫瑰糠疹临床观察

目的 观察凉血解毒汤联合窄谱中波紫外线照射治疗玫瑰糠疹的临床疗效。 方法 将82例玫瑰糠疹患者随机分为治疗组43例和对照组39例,治疗组口服自拟凉血解毒汤和照射窄谱中波紫外线,对照组仅口服自拟凉血解毒汤,治疗1个月后观察两组玫瑰糠疹严重程度评分（pityriasis rosea severity score,PRSS）,皮疹止痒、停发、消退时间和临床疗效。结果 两组患者治疗后PRSS均显著减少（P＜0.05）;两组PRSS差值比较,差异无统计学意义（P＞0.05）。两组患者皮疹止痒时间、皮疹停发时间、皮疹消退时间比较,差异均有统计学意义（P＜0.05）。治疗组临床疗效优于对照组（P＜0.05）。结论 口服中药联合窄谱中波紫外线照射治疗玫瑰糠疹,疗效优于单用口服中药且不良反应小。     

新藤黄酸对黑色素瘤B16细胞凋亡的作用

目的 探讨新藤黄酸（gambogenic acid,GNA）对小鼠黑色素瘤B16细胞凋亡的影响。方法 用不同浓度的GNA培养B16细胞24 h,倒置显微镜下观察细胞形态;采用四甲基偶氮唑蓝(methyl thiazolyl tetrazolium,MTT)法测定GNA对B16细胞增殖的抑制作用;荧光显微镜观察吖啶橙/溴化乙锭（acridine orange/ethidium bromide,AO/EB）染色后细胞凋亡;采用膜联蛋白Ⅴ-异硫氰酸荧光素/碘化丙啶双染,流式细胞仪检测B16细胞凋亡。结果 GNA作用B16细胞后,细胞的抑制率随着药物浓度的增加而升高;AO/EB染色荧光显微镜观察显示GNA处理的B16细胞出现了典型的凋亡特征;流式细胞仪检测显示GNA处理的B16细胞随着药物浓度的增大凋亡率随之上升。结论 GNA在一定的范围内,能够浓度和时间依赖性地通过诱导细胞凋亡抑制小鼠黑色素瘤B16细胞的增殖。     

叶下珠水提物对裸鼠人肝癌移植瘤HBx和VEGFR3表达的影响

目的 探讨乙型肝炎病毒x基因（hepatitis B virux x gene, HBx）和血管内皮生长因子受体3（vascular endothelial growth factor receptor 3, VEGFR3）基因与乙型肝炎相关肝癌的相关性,以及叶下株水提物（aqueous extract of phyllanthus urinaria L, AEP）对HBx和VEGFR3蛋白表达的干预作用。方法 将60只Balb/c-nu裸鼠随机分为10组。分别复制转染HBx、氯霉素乙酞转移酶（chloramphenicol acetyltransferase,CAT）和VEGFR3基因的HepG2肝癌细胞的裸鼠皮下移植瘤模型,用AEP进行干预,以生理盐水、环磷酰胺（cyclophosphamide,CTX）作为对照,共治疗6周。观察模型复制后不同时间各组裸鼠的移植瘤生长状况,采用酶联免疫吸附试验检测各组裸鼠血清甲胎蛋白（alpha fetal protein, AFP）水平,采用Western blot方法检测移植瘤组织中HBx和VEGFR3蛋白表达水平。结果 肝癌移植瘤模型复制成功。各组裸鼠体质量均随着时间显著增长。实验结束时,接种HepG2-HBx细胞的各组中,AEP处理组肿瘤质量显著低于CTX处理组（P＜0.05）,其AFP水平显著高于NS处理组（P<0.05）,其移植瘤组织中HBx表达水平显著低于NS组（P<0.05）。接种相同肝癌细胞的各组中,AEP处理组VEGFR3表达水平最低,但与CTX组VEGFR3表达水平比较,差异无统计学意义（P＞0.05）;AEP组和CTX组VEGFR3表达水平显著低于NS组（P＜0.05）。对于相同的处理因素,接种HepG-HBx细胞和HepG-CAT细胞的裸鼠移植瘤组织中VEGFR3表达水平均显著低于接种HepG-VEGFR3细胞的裸鼠（P＜0.01）。结论 AEP通过直接抑制HBx蛋白和VEGFR3蛋白表达,及通过抑制HBx蛋白表达而间接抑制VEGFR3蛋白表达,达到对肝癌移植瘤生长的抑制作用。