蛋鸡输卵管中与精子储存相关四种酶分布的组织化学染色观察

应用组织化学方法在组织和细胞水平上对碱性磷酸酶、酸性磷酸酶、过氧化氢酶以及三磷酸腺苷酶在蛋鸡输卵管内的分布特点及分布意义进行了研究分析。结果显示,4种酶在蛋鸡输卵管各段的分布部位以及反应强弱存在差异。碱性磷酸酶在子宫部分布最多,在有精子储存的黏膜固有层腺体和血管周围也有明显分布;酸性磷酸酶在阴道子宫交界部分布最多,酸性磷酸酶阳性反应呈颗粒状,主要分布于黏膜上皮和固有层腺体;过氧化氢酶在输卵管各段血管周围以及子宫阴道交界处精子腺和蛋白分泌部反应明显;三磷酸腺苷酶在各段尤其是子宫阴道交界处精子腺部位黏膜上皮纤毛和腺体分布较多,血管也有一定分布。表明这些酶在精子储存时对精子的活性和储存环境的维持、物质运输及分泌等活动具有一定的生物学作用。     

XPG基因表达用于法医学年龄推断

目的探讨着色性干皮病G组(xeroderma pigmentosum group G,XPG)基因在不同年龄段健康汉族人群中的表达情况,分析XPG mRNA和蛋白表达量与年龄之间的相关性,以期为法医学年龄推断提供新的分子生物学指标。方法收集150名不同年龄段健康汉族人的外周血样,采用TRIzol法提取外周血单个核细胞(peripheral blood mononuclear cell,PBMC)总RNA,通过实时荧光定量PCR检测XPG mRNA在PBMC的相对表达量,酶联免疫吸附试验检测XPG蛋白在血浆中的表达量。结果 XPG mRNA及其蛋白表达量在≤18岁组与19~45岁组之间、≤18岁组与≥46岁组之间的差异均有统计学意义(P0.05),但19~45岁组与≥46岁组之间的差异无统计学意义(P0.05)。XPG mRNA和蛋白表达量均无性别差异(P0.05)。结论 XPG mRNA在PBMC的相对表达量在低龄段内随年龄增加而下降,其血浆中蛋白随年龄增加而升高;XPG基因有望成为法医学年龄推断的新型指标之一。     

单管一步甲基化可变位点分析技术

目的建立单管一步甲基化可变位点（methylationvariableposition,MVP）分析技术一单管消化后PCR链融解曲线分析（post—digestionPCR—meltingcurveanalysis,PDP—MCA）。方法以文献报道的差异甲基化区（differentiallymethylatedregion,DMR）为模型,在MVP两侧设计一组解链温度各不相同的引物。应用FastDigest甲基化敏感性限制酶（methylation—sensitiverestrictionenzyme,MSRE）,在同一反应管内顺次进行DNA的酶切、复合扩增和MCA检测．生成MCA图谱。同时用该方法和传统的MSRE—PCRMCA技术检测相同样品（外周静脉血、精液、阴道液各5份）,比较两种方法检测结果,验证其可行性,并分析比较不同样品的MCA／HRM图谱。结果解链温度相差2℃以上的片段,MCA峰分离良好,复合扩增后可以用MCA技术检测。应用单管PDP—MCA技术,可以集酶切、扩增和检测三步于一管,在2h内得到与传统方法一致的特异性图谱和数据,并实现样品的快速分类鉴别。结论单管PDP-MCA技术可以实现多个MVP的单管、闭管检测,具有简便、快速、易于自动化等优点,可用于样品DNA甲基化差异的检测。     

Evaluation of Tamm‐Horsfall Protein and Uroplakin III for Forensic Identification of Urine

Abstract: In this study, Tamm‐Horsfall protein (THP), a major component of urinary protein, and uroplakin III (UPIII), a transmembrane protein widely regarded as a urothelium‐specific marker, were evaluated for forensic identification of urine by ELISA and/or immunohistochemistry. THP was detected in urine, but not in plasma, saliva, semen, vaginal fluid, or sweat by the simple ELISA method developed in this study. In addition, most aged urine stains showed positive results. The urine specificity of THP was confirmed by gene expression analysis. Therefore, as reported previously, ELISA detection of THP can be used as a presumptive test for urine identification. UPIII was specific for immunohistochemical staining of cells in centrifuged precipitate of urine. However, ELISA and RT‐PCR for UPIII were not specific for urine. UPIII may be applicable for forensic urine identification by immunohistochemistry.     

应用酶联免疫吸附试验检测实验感染绵羊进行性肺炎病毒的山羊血清抗体

用酶联免疫吸附试验（ＥＬＩＳＡ）在接种绵羊进行性肺炎病毒（ＯＰＰＶ）的山羊血清中可测到ＯＰＰＶ抗体。抗体的最早检出时间是接毒后的第１５ｄ，且４只接毒山羊都呈阳性反应。检测结果表明，ＯＰＰＶ可在山羊体内诱生较强的体液免疫应答反应。试验所用的ＥＬＩＳＡ除可用于ＯＰＰＶ通过山羊体的传代研究之外，还可应用于绵羊进行性肺炎（ＯＰＰ）的诊断     

异马钱子碱对心肌细胞作用的单钙通道及透射电镜分析

用膜片钳的方法，在培养ｗｉｓｔａｒ大鼠乳鼠心室肌细胞上分别记录Ｔ型，Ｌ型，Ｂ型钙通道的单通道活动。异马钱子碱显著激动３种钙通道的活动，使其开放时间延长，关闭时间缩短，开放概率地加，而对通过每一种离子通道的离子流幅值无明显影响。采用电镜观察的方法，结果表明异马钱子碱能明显地抵消黄嘌呤黄嘌呤氧化酶引起的破坏培养的心室肌细胞肌丝和线粒体等超微结构的作用。     

用甘肃棘豆草粉饲喂苦马豆素-BSA免疫山羊的血清相关酶分析

用甘肃棘豆草粉饲喂经苦马豆素(SW)-BSA免疫接种的山羊,检测分析了山羊血清中与中毒相关的酶的活性,以探讨SW-BSA免疫接种对动物机体组织器官的保护作用。将24只山羊随机分为免疫对照组、免疫攻毒组和未免疫攻毒对照组,免疫对照组和免疫攻毒组山羊接种SW-BSA,免疫攻毒组和未免疫攻毒对照组山羊按干重拌料饲喂10 g/(kg.d)甘肃棘豆草粉,检测血清谷草转氨酶(GOT)、谷丙转氨酶(GPT)、乳酸脱氢酶(LDH)、碱性磷酸酶(AKP)、尿素氮(BUN)和α-甘露糖苷酶(AMA)的变化。结果表明,免疫攻毒组山羊较未免疫攻毒对照组山羊LDH活性升高延缓28 d,AKP活性升高延缓14 d,AMA活性降低延缓21 d,BUN活性升高延缓14 d,GOT活性升高延缓28 d。这些酶活性延缓变化,说明SW-BSA免疫动物后,在甘肃棘豆攻毒后的30 d内,能够有效地延缓SW对山羊肝、心和肾等组织器官的损伤。     

Parent and Metabolite Opioid Drug Concentrations in Unintentional Deaths Involving Opioid and Benzodiazepine Combinations,,

Effects of benzodiazepines on postmortem opioid parent and parent/metabolite blood concentration ratios were determined for fentanyl‐, hydrocodone‐, methadone‐, or oxycodone‐related accidental deaths. These opioids are partially metabolized by the CYP3A4 enzyme system, which is also affected by diazepam and alprazolam. Opioid/metabolite combinations examined were as follows: fentanyl/norfentanyl, hydrocodone/dihydrocodeine, methadone/EDDP, and oxycodone/oxymorphone. Parent opioid concentrations were analyzed for 877 deaths. Parent/metabolite concentration ratios were analyzed for 349 deaths, excluding cases with co‐intoxicants present known to interfere with opioid elimination. Alprazolam in combination with diazepam significantly decreased median hydrocodone concentrations by 48% (p = 0.01) compared to hydrocodone alone. The methadone parent/metabolite concentration ratio was reduced by 35% in the presence of diazepam compared to methadone alone (p = 0.03). Benzodiazepines did not statistically significantly affect fentanyl or oxycodone concentrations. Possible factors affecting opioid concentrations and possible toxicity development, including any differential effects on specific opioids, should continue to be explored.     

禽分枝杆菌副结核亚种保护性抗原的研究进展

为了给副结核病新型疫苗研究中抗原的选择提供参考,综述了超氧化物歧化酶和巯基过氧化物酶等酶类、热休克蛋白、抗原85复合物及其他一些抗原在激发实验动物细胞和体液免疫中所发挥的作用,同时总结了在实际应用中可供借鉴的经验。     

Gastrointestinal Functionality of Aquatic Animal (Oreochromis niloticus) Carcass in Water Allows Estimating Time of Death

Postmortem changes have been previously studied in some terrestrial animal models, but no prior information is available on aquatic species. Gastrointestinal functionality was investigated in terms of indices, protein concentration, digestive enzyme activity, and scavenging activity, in an aquatic animal model, Nile tilapia, to assess the postmortem changes. Dead fish were floated indoors, and samples were collected within 48 h after death. Stomasomatic index decreased with postmortem time and correlated positively with protein, pepsin‐specific activity, and stomach scavenging activity. Also intestosomatic index decreased significantly and correlated positively with protein, specific activity of trypsin, chymotrypsin, amylase, lipase, and intestinal scavenging activity. In their postmortem changes, the digestive enzymes exhibited earlier lipid degradation than carbohydrate or protein. The intestine changed more rapidly than the stomach. The findings suggest that the postmortem changes of gastrointestinal functionality can serve as primary data for the estimation of time of death of an aquatic animal.