双重实时荧光PCR快速鉴别人参和西洋参＊

目的 建立一种能够快速鉴定人参和西洋参的双重实时荧光PCR方法。方法 通过对人参属及其近似种基因序列的分析比对，设计特异性的引物探针，建立双重实时荧光PCR检测方法。PCR反应体系为Premix Ex Taq （Probe qPCR） 10 μL，人参上下游引物各0.5 μL，西洋参上下游引物各0.3 μL，人参与西洋参特异性探针各0.4 μL，DNA模板2 μL，灭菌去离子水补至20 μL。反应条件为95℃预变性30 s；然后以95℃变性5 s，60℃退火延伸30 s进行40个循环。应用该方法测试了27份DNA样品，包括7份人参、6份西洋参、4份人参与西洋参混合样、10份其他人参属样品及其他常见中药材样品的DNA，以确定该方法的特异性。将人参与西洋参样品DNA混合后10倍稀释成不同浓度后进行检测，用以确定该方法的灵敏度。结果 人参及西洋参样品均在特定的荧光通道下出现典型的阳性扩增曲线，人参与西洋参混合样品均同时出现两条阳性扩增曲线，其它对照样品及空白对照均没有出现阳性扩增曲线。灵敏度检测结果显示该方法检测人参及西洋参的最低检测限均为2 pg DNA/反应。结论 本实验建立的双重实时荧光PCR方法能够同时快速、准确、灵敏地鉴别出人参和西洋参。     

一种快速检测HBV基因的荧光定量PCR法的建立

目的：利用二聚体蝎型荧光探针技术,建立一种能用于临床的快捷、敏感、特异、价格低廉的HBV实时荧光定量PCR检测方法。方法：根据HBV基因组保守区域precore／core设计二聚体蝎型荧光探针和引物,构建质粒标准品,优化荧光定量PCR条件,并进行方法学评价。结果：所建方法的检测范围为10^1～10^8 copies／μl,灵敏度达到10copies／μl,低拷贝样品的批内和日间重复性（CV）分别为1．71％和2．57％,高拷贝样品的批内和日间GV,分别为3．00％和3．86％。与商品化TaqMan试剂盒相比,本法阳性检出率较高,且检测时间缩短了45min（约减少1／3）,成本降低50％。结论：成功建立了快速检测HBVDNA的二聚体蝎型荧光定量PCR方法,为研发适合临床应用的HBV基因定量检测试剂盒奠定了基础。     

Duplex sonographic criteria for measuring carotid stenoses

PURPOSE: The aim of this retrospective study was to determine optimal duplex sonographic criteria for use in our institution for diagnosing severe carotid stenoses and to correlate those findings with angiographic measurements obtained by the European Carotid Surgery Trial (ECST), North American Symptomatic Carotid Endarterectomy Trial (NASCET), and Common Carotid (CC) methods of grading carotid stenoses. METHODS: We analyzed the angiographic data using the ECST, NASCET, and CC methods and compared the results with the duplex sonographic findings. We then calculated the sensitivity, specificity, positive and negative predictive values, and accuracy of the duplex sonographic method. Taking these parameters into account, the optimal intrastenotic peak systolic velocity (PSV) and end diastolic velocity (EDV) were derived for diagnosing severe stenoses according to the 3 angiographic methods. RESULTS: Optimal PSV and EDV values for diagnosing a 70% or greater stenosis in our laboratory were as follows: with the NASCET method of angiographic grading of stenoses, PSV 220 cm/second or greater and EDV 80 cm/second or greater, and with the ECST and CC methods, PSV 190 cm/second or greater, and EDV 65 cm/second or greater. The optimal PSV and EDV for diagnosing a stenosis of 80% or greater with the ECST grading method were 215 cm/second or greater and 90 cm/second or greater, respectively. CONCLUSIONS: Duplex sonography is a sensitive and accurate tool for evaluating severe carotid stenoses. Optimal PSVs and EDVs vary according to the angiographic method used to grade the stenosis. They are similar for stenoses 70% or greater with the NASCET method and for stenoses 80% or greater with the ECST method.     

Hemodynamic evaluation of the female pelvic vessels using a high-frequency transvaginal image-directed Doppler system

Perfusion characteristics of the female pelvic vessels were studied by a high-frequency transvaginal image-directed Doppler system. A 5-MHz Doppler transducer was coupled to a 6.5-MHz probe especially designed for intravaginal use. Distinct patterns of flow velocity waveforms were obtained from the ascending branch of the uterine artery, before and during pregnancy. A gradual increase in diastolic flow throughout gestation was observed, consistent with a decrease in vessel resistance. Flow velocity profiles in the ovarian artery were sampled from the infundibulo-pelvic ligament. Only limited diastolic flow could be detected in either ovary during the follicular phase of the cycle. In the luteal phase there was a marked increase in this flow but only in the ovary containing the corpus luteum. In early pregnancy the high diastolic flow was maintained or even increased, consistent with a low arterial resistance. High image resolution and a higher frequency Doppler transducer increase the usefulness of this technique in the hemodynamic evaluation of the female pelvic vessels.     

尿激酶改善长时间热缺血肾脏功能的实验研究

目的 探讨尿激酶对长时间热缺血(warm ischemia,WI)肾脏的形态和功能恢复的影响.方法 30只家兔按随机数字表法分为3组:假手术、热缺血组(WI组)、热缺血+尿激酶治疗组(WI+UK组).夹闭左侧肾动、静脉90 min模拟热缺血过程,HCA液冷灌注及肾脏血流复灌,模拟自体原位肾移植.比较各组血肌酐(Cr)、肾脏组织病理改变(HE、Masson染色)和免疫组化检测血栓调节蛋白(thrombomodulin,TM).结果 与假手术组比较,WI组和WI+UK组术后Cr水平明显升高(P＜0.05),术后35 d Cr水平WI+UK组[(118.99±17.18) μmol/L]明显低于WI组[(165.34±8.38) μmol/L,P＜0.01].WI组肾小球内大量微血栓形成,肾小管水肿、坏死;WI+UK组肾小球未见血栓,肾小管未见异常.WI组TM表达明显高于假手术组(P＜0.01),WI+ UK组明显低于WI组(P＜0.05).结论 热缺血90 min后有明显的微血栓形成,导致肾脏结构破坏及肾功能下降.尿激酶治疗后微血栓溶解,有利于肾脏结构和功能恢复.     

Renal pelvis urothelial carcinoma of the upper moiety in complete right renal duplex: a case report

Urothelial carcinoma (UC) originated from renal pelvis is the common tumor of the urinary system, however, neoplasia of the renal pelvis in duplex kidneys is extremely rare, especially in the complete renal and ureteral duplex cases. We present the first case of renal pelvis UC of the upper moiety in a complete right renal duplex. This male patient has bilateral complete renal and ureteral duplex. To the best of our knowledge, this is the first reported case of renal pelvis UC in a complete renal duplex system. After this experience we feel that the diagnosis of renal pelvis UC in duplex kidneys is not so easy, and once the diagnosis is determined, the whole renal duplex units and bladder cuff or ectopic orifice should be excised radically.