Analysis of cyclosporine A and its metabolites in rat urine and feces by liquid chromatography–tandem mass spectrometry

A sensitive and specific liquid chromatography–tandem mass spectrometry (LC–MS/MS) method was developed and validated for the quantification of cyclosporine A (CyA) and the identification of its metabolites in rat urine and feces. The analytes were extracted from waste samples via liquid–liquid extraction. A Turboionspray source was used as a detector. It was operated in a positive ion mode with transitions of m/z 1225 → m/z 1112 for CyA and in a selected multiple reactions monitoring (MRM) mode with transitions of m/z 1239 → m/z 1099 for the internal standard (cyclosporine D, CyD). Linear calibration curves were obtained for CyA concentration ranges of 12.5–250 ng mL^?1 in urine and 2.5–375 ng mg^?1 in feces. The intra- and inter-day precision values (relative standard deviation) obtained were less than 8%, and the accuracy was within ±15% for each of the analytes. Extraction recoveries of CyA and CyD were both over 80%. The identification of the metabolites and elucidation of their structure were performed on the basis of their retention times and mass spectrometry fragmentation behaviors. A total of seven metabolites in rat feces were identified as dimethyl CyA, hydroxy CyA, and dihydroxy CyA after the oral administration of cyclosporine A-Eudragit^® S100 nanoparticles (CyA-NP). Six of these metabolites were also detected in rat urine. A possible metabolic pathway was also proposed. The newly developed method was proven to be sensitive, simple, reproducible, and suitable for the rapid determination of CyA. It was successfully employed to study the excretion of CyA in rats and could be used to better understand the in vivo metabolism of CyA-NP, a potentially effective nanoparticle system.     

基于相对密码子频率的芜菁花叶病毒基因组序列的进化分析

现有92株芜菁花叶病毒(TuMV)的全基因组序列已在GenBank报道,据分析报道其中58株不含重组序列。利用系统聚类法对92株TuMV的全基因组序列和58株TuMV全基因组序列的相对密码子频率RSCU值进行聚类分析。同时利用系统发育分析方法分析了这92株和58株TuMV全基因组序列。结果发现,92株芜菁花叶病毒株的密码子偏性聚类树与其系统进化树的一致度很低;而不含重组序列的58株芜菁花叶病毒株的密码子偏性聚类树与其系统进化树的一致度却非常高,且与寄生宿主类型基本对应。这表明在不存在重组的情况下,TuMV密码子频率的偏性可能是宿主内的一种选择压力,影响TuMV基因组的点突变进化方向,促使TuMV适应宿主内环境。     

越香竹——香竹属一新种

报导了从云南省金平县用种子(种子来源于越南莱州省的燕汤)繁殖的幼苗引入四川长宁县栽培的香竹属一新种越香竹(Chimonoca lamus peregrinus Yiet L.S.Ma)。新种与马关香竹(Ch.makuanensis Hsuehet Yi)相近似[1,3～5],但节间幼时微被灰粉,密被紫色小点,在分枝一侧中下部或基部扁平,并具纵脊和沟槽,无毛,刺状气生根较短,长约1～3mm,箨鞘短于节间长度,无不同颜色的纵条纹,箨舌三角形、"山"字形或稀近于截平形,高1～2mm,小枝具叶(4)6～8枚,叶鞘口繸毛2～4枚,长3～4mm,叶片下面淡绿色;也近似角香竹(Ch.bicorniculatusS.F.LietZ.P.Wang)[2,4,5],不同在于节间无毛,在分枝一侧中下部或基部扁平,并具纵脊与沟槽,节上气生根刺离生,箨鞘顶端两侧低于或远低于中部,箨舌高1～2mm,小枝具叶(4)6～8枚,叶片长8.5～13cm,宽6～10mm,次脉3(4)对,笋期在春季和秋季,易于区别。     

循环肿瘤细胞在乳腺癌预后评价中的应用和意义

循环肿瘤细胞(CTCs)是导致肿瘤转移的主要原因,它在外周血中的含量与乳腺癌的转移、治疗和预后有密切的关系.本文对于CTCs的概念、检测方法、以及作为肿瘤转移预测指标的优势进行了综述.并对其在临床上的运用前景进行了展望.     

Ⅱ期经皮输尿管镜治疗输尿管上段结石梗阻致孤立肾急性肾功能衰竭

目的:探讨经皮肾微造瘘术后Ⅱ期经皮输尿管镜治疗输尿管上段结石致孤立肾急性肾功能衰竭的安全性与有效性。方法:从2004年7月～2009年5月,利用经皮肾微造瘘建立经皮肾通道,引流1周后肾功能明显好转,再行经皮肾输尿管镜碎石治疗孤立肾输尿管上段结石。结果:16例患者中,所有患者均为单通道取石,结石清除率例(81.2%),未出现高热、出血等并发症。术后1月复查13例无结石残留。结论:微创经皮输尿管镜分期治疗输尿管上段结石致孤立肾急性肾功能衰竭是安全、有效的,同传统经皮肾镜相比,具有对病人创伤小,易恢复等优点。     

隐形针灸治疗慢性疲劳综合征的临床疗效观察

目的:观察隐形针灸对慢性疲劳综合征的疗效及作用机制初步探讨.方法:56例慢性疲劳综合征患者采用隐形针灸进行治疗,通过采用慢性疲劳综合征的疗效标准和疲劳量袁(FS)来观察患者治疗前、后病症状变化,进行临床疗效评价.结果:隐形针灸治疗慢性疲劳综合征治疗3个疗程后,患者的症状明显减轻或消失,总有效率达91.1%.结论:隐形针灸治疗慢性疲劳综合征是一种有效的治疗方法.     

Hypomania: a transcultural perspective

This study examined the transcultural robustness of a screening instrument for hypomania, the Hypomania Checklist-32, first revised version (HCL-32 R1). It was carried out in 2606 patients from twelve countries in five geographic regions (Northern, Southern and Eastern Europe, South America and East Asia). In addition, GAMIAN Europe contributed data from its members. Exploratory and confirmatory factor analyses were used to examine the transregional stability of the measurement properties of the HCL-32 R1, including the influence of sex and age as covariates. Across cultures, a two-factor structure was confirmed: the first factor (F1) reflected the more positive aspects of hypomania (being more active, elated, self-confident, and cogni-tively enhanced); the second factor (F2) reflected the more negative aspects (being irritable, impulsive, careless, more substance use). The measurement properties of the HCL-32 R1 were largely invariant across cultures. Only few items showed transcultural differences in their relation to hypomania as measured by the test. F2 was higher among men and in more severe manic syndromes; F1 was highest in North and East Europe and lowest in South America. The scores decreased slightly with age. The frequency of the 32 items showed remarkable similarities across geographic areas, with two excep-tions: South Europeans had lower symptom frequencies in general and East Europeans higher rates of substance use. These findings support the interna-tional applicability of the HCL-32 R1 as a screening instrument for hypomania.     

獐牙菜小孢子发生及雄配子体发育(英文)

用石蜡切片法对獐牙菜小孢子发生及雄配子体发育过程进行首次观察研究。主要结果如下:花药四室,药壁发育为基本型;绒毡层异型起源,属于腺质型绒毡层,药室内具有的退化绒毡层核是早期该层细胞有丝分裂凸入药室中央并原位退化形成的;中层细胞2层;药室内壁同表皮同时宿存,细胞柱状伸长,纤维状加厚。小孢子母细胞减数分裂为同时型,四分体排列方式主要为四面体形,少数为左右对称形和十字交叉形;成熟花粉多为2-细胞类型,偶见3-细胞型,具三萌发孔。     

表达血管内皮生长因子-siRNA及双自杀基因yCDglyTK的联合基因载体构建及其应用研究

构建了新型联合基因载体pcDNA3.1(-)VEGF-siRNA/yCDglyTK,研究其在人胃癌细胞系SGC7901细胞中的表达和杀伤作用.构建靶向血管内皮生长因子(VEGF)的干扰质粒pGenesil-VEGF-siRNA,采用PCR法从中扩增siRNA表达框(含U6启动子),亚克隆至双自杀基因载体pcDNA3.1(-)CV-yCDglyTK,构建联合基因质粒pcDNA3.1(-)VEGF-siRNA/yCDglyTK;通过酶切、测序等鉴定重组质粒;以磷酸钙纳米颗粒为载体,将干扰质粒、双自杀基因质粒及联合基因质粒转染SGC7901细胞,RT-PCR、Western-blot验证目的基因表达;MTT法检测转染细胞对5-氟胞嘧啶(5-FC)的敏感性.结果表明:酶切及测序证实联合基因载体pcDNA3.1(-)VEGF-siRNA/yCDglyTK构建成功;SGC7901细胞转染联合基因质粒后,RT-PCR、Western-blot证实融合自杀基因表达,而VEGF基因表达下调;在前体药物5-FC作用下,转染联合基因组细胞存活率最低,与其他组比较有统计学差异.成功构建联合基因载体pcDNA3.1(-)VEGF-si...     

‘红巴梨’果皮UFGT基因的克隆及表达分析

以‘巴梨’红色芽变品种‘红巴梨’果皮为材料,采用同源克隆技术和RACE结合的方法,克隆了UFGT(UDP-葡萄糖:类黄酮3-O-葡萄糖基转移酶)蛋白的部分cDNA,命名为Pc UFGT。结果表明:该cDNA片段长为1 089 bp,与苹果UFGT基因序列一致性达89%,氨基酸序列同源性为85%,含有糖基转移酶的UDPGT、COG1819和MGT等保守域。荧光实时定量PCR分析表明,该基因在‘红巴梨’幼果期表达强度约为‘巴梨’的2倍,而果实成熟期表达强度略低于‘巴梨’;该基因在‘红巴梨’果肉中不表达。