液质联用法测定龙胆水煎剂中三种环烯醚萜的含量

目的利用液质联用（LC／MS／MS）方法测定龙胆水煎剂中环烯醚萜类化合物龙胆苦苷、獐牙菜苦苷和马钱苷的含量。方法色谱柱为J’Sphere ODS-H80（150mm×2．0mm，4μm）；流动相为甲醇-10mmol·L^-1醋酸铵溶液（30：70）；流速为200μl／min。样品经电喷雾离子源（ESI）正离子化后，通过三级四极杆串联质谱仪，采用多反应离子监测方式测定样品中龙胆苦苷（m／z 357．2-195．3）、獐牙菜苦苷（m／z 392．1→195．2）和马钱苷（m／z 408．4→229．1）浓度。结果在该分析条件下，3种化合物能够同时进行检测。龙胆苦苷浓度在200～5000ng·ml^-1范围内线性关系良好，回归方程为Y=1．26×10^3X＋2．27×10^3，r=0．9983；獐牙菜苦苷浓度在20-500ng·ml^-1范围内线性关系良好，回归方程为Y=360X＋144，r=0．9991；马钱苷浓度在20-500ng·ml^-1范围内线性关系良好，回归方程为Y=782X＋42．6，r=0．9994。对照品峰面积精密度RSD分别为龙胆苦苷2．52％，獐牙菜苦苷2．77％和马钱苷2．89％；平均加样回收率分别为龙胆苦苷98．2％，獐牙菜苦苷99．1％和马钱苷99．7％。结论该方法灵敏、准确、特异性强。     

舒尼替尼和雷米普利联合用药在大鼠体内的药代动力学相互作用

初步考察舒尼替尼和雷米普利联合用药在大鼠体内的药代动力学相互作用。18只雄性SD大鼠随机分成3组,分别灌胃给予舒尼替尼、雷米普利、舒尼替尼合用雷米普利,连续给药10 d,分别于给药第1天和第10天后不同时间点采集血样,用LC-MS/MS测定雷米普利拉和舒尼替尼的血药浓度,并计算二者药代动力学参数。单剂量联合用药时,与单用雷米普利相比,合用舒尼替尼后,雷米普利拉t_max显著减小,t_1/2显著增加,AUC_0-∞没有显著性差异,表明其吸收加快,消除减慢,但整体吸收程度不变。多剂量给药后,联合用药组雷米普利拉CL显著降低、AUC_0-∞显著增大,表明联合用药导致雷米普利拉在大鼠体内消除减慢,并有明显的蓄积。单剂量和多剂量联合用药后舒尼替尼的药代动力学行为都没有发生显著变化。结果显示,舒尼替尼和雷米普利联合用药后会导致雷米普利拉消除减慢,并且单剂量联合用药后其吸收变快,多剂量联合用药后其在体内有明显蓄积,提示二者存在一定的药代动力学相互作用。     

Cognition in the early stage of multiple sclerosis

Objective Cognitive dysfunctions may contribute to limitation of everyday activities of patients with multiple sclerosis (MS). Recent studies have demonstrated that 45 to 65% of MS-patients are cognitively impaired. The profile of MS-related cognitive dysfunctions varies greatly. It includes memory and learning deficits, attention deficits, executive dysfunctions and visuo-spatial deficits. Most studies of cognition in MS examined patients in later stages, often including MS-patients with marked physical disabilities. Studies of cognitive dysfunctions in the early stage of the disease are rare. This study specifically aimed at evaluating and characterizing cognitive impairments in the early stage of MS, and determining specific patterns of cognitive dysfunction. Methods 21 MS patients, experiencing their first neurological symptoms not more than two years previously, and 22 healthy controls were compared. A comprehensive neuropsychological test-battery was used to evaluate MS-related cognition. The battery consisted of memory and learning tests, executive functioning tests and a visuo spatial functioning test. A computerized attention test-battery was also included, which assess accuracy and speed of test responses. In addition depression and intellectual capabilities were assessed. Results Compared with healthy controls, MS-patients in the early stage of the disease performed significantly lower on each neuropsychological assessment, except for verbal short-term memory. In particular, MS-patients showed a lengthened reaction time for simple and focused attention (19–38%), impaired non-verbal memory function (RVDLT recognition: 33%) and a planning deficit (24%). Associations between information processing speed and disease course and the employment situation were additionally found. However, patients did not have clinically relevant depression rates on the ADS-L and visuo spatial abilities remain preserved. Conclusion Our findings revealed discrete cognitive dysfunction in MS-patients within the early stage of the disease. Received in revised form: 18 January 2006     

65例多发性硬化患者的临床分析

目的总结多发性硬化(MS)患者的临床特点(精神情感障碍)以及脑脊液、诱发电位、影像学改变进行分析。方法回顾分析65例MS患者的有关临床和实验室资料。结果65例中,男17例,女48例;年龄16~68岁,平均(36.5±14.1)岁,男女比为1∶2.8。MS首发及常见症状为感觉异常、肢体无力、视力减退及括约肌功能障碍,病变部位以脊髓受累最多见,部分患者存在精神情感障碍。结论出现感觉异常有助于MS早期诊断,诱发电位及MRI有助于发现亚临床病变;括约肌功能异常多见,可能与脊髓受累较多有关;MS患者的精神情感应受人们关注。     

1,25 Dihydroxyvitamin-D3 modulates JAK-STAT pathway in IL-12/IFNgamma axis leading to Th1 response in experimental allergic encephalomyelitis

Experimental allergic encephalomyelitis (EAE) is a Th1 cell-mediated autoimmune disease model of multiple sclerosis (MS). Vitamin D deficiency is commonly observed in MS patients and vitamin D supplements reduce the clinical symptoms of EAE and MS. Earlier studies have shown that in vivo treatment with vitamin D analogs ameliorates EAE in association with the inhibition of IL-12 production and Th1 differentiation. The mechanisms in the regulation of Th1 response by vitamin D in EAE/MS are, however, not known. We show that in vivo treatment of C57BL/6 and SJL/J mice (i.p.) with 100 ng of 1,25 dihydroxyvitamin D3, on every other day from Day 0-30, ameliorates EAE in association with the inhibition of IL-12 production and neural antigen-specific Th1 response. In vitro treatment with 1,25(OH)2D3 inhibited IFNgamma-induced tyrosine phosphorylation of STAT1, without affecting JAK2, in EOC-20 microglial cells. Treatment of activated T cells with 1,25(OH)2D3 also inhibited the IL-12-induced tyrosine phosphorylation of JAK2, TYK2, STAT3, and STAT4 in association with a decrease in T cell proliferation in vitro. These findings highlight the fact that vitamin D modulates JAK-STAT signaling pathway in IL-12/IFNgamma axis leading to Th1 differentiation and further suggest its use in the treatment of MS and other Th1 cell-mediated autoimmune diseases.     

Proton magnetic resonance spectroscopy of normal appearing white matter in asymptomatic relatives of multiple sclerosis patients

Proton magnetic resonance spectroscopy of normal appearing white matter (NAWM) was performed in 25 first-degree relatives of patients with multiple sclerosis (MS). In relatives of MS patients, insignificant trends in the NAA/Cho ratio (to be lower) and in the Cho/Cr ratio (to be higher) were found when compared with healthy controls. These results demonstrated that there are only minimal, insignificant changes in commonly identified metabolite concentrations in NAWM of relatives of MS patients when compared with healthy subjects.     

蒸煮鸡肉的挥发性香气成分

采用同时蒸馏萃取法(即SimultaneousDistillationExtractor,以下简称SDE)提取蒸煮鸡肉中的挥发性香气成分,并采用气相色谱质谱联用法(GC-MS)对香气成分进行了鉴定.结果得到化合物44种,共分成9类,其中羰基化合物最多、芳香族次之.羰基化合物对鸡肉特征香气的形成起重要作用,芳香族和呋喃是蒸煮鸡肉的主要香气成分.     

辛弗林对CUMS大鼠抑郁样行为和血清代谢组学的影响

目的 通过观察分析大鼠慢性应激引起的行为学和血清代谢组变化以及辛弗林抗抑郁的有效治疗,发现潜在的抗抑郁新药先导化合物和生物标志物。方法 将24只大鼠分为正常组、模型组和辛弗林组。模型组为6周慢性温和不可预知应激(CUMS)模型大鼠,用体质量变化、糖水偏好、开场实验作为抑郁评估指标,后2周辛弗林(Syn 20 mg/kg)干预治疗后采集大鼠血清进行气相色谱-质谱联用(GC-MS/MS)测定,研究CUMS大鼠血清内源性代谢物变化,以及辛弗林的干预作用,寻找其抗抑郁潜在的先导化合物和生物标志物,并找到相关靶向代谢通路。结果 辛弗林干预后,在增加CUMS大鼠体质量、提高糖水偏好率、增加开场运动总距离和理毛次数的同时,还能逆转CUMS大鼠血清中磷酸乙醇胺、苯甲酸、胆固醇、谷氨酸等14种代谢物的变化使其趋于正常组,并发现谷氨酰胺和谷氨酸的代谢、缬氨酸和异亮氨酸的生物合成等9条主要牵涉辛弗林的靶向代谢通路。结论 辛弗林可能通过调节谷氨酰胺和谷氨酸的代谢、缬氨酸和异亮氨酸的生物合成等生物学途径发挥对CUMS大鼠抑郁样行为的调正作用。      

生物质谱技术在腺相关病毒（AAV）载体制剂质量控制中的应用

腺相关病毒（adeno-associated virus,AAV）是一种基因治疗中常用的病毒载体。由于其安全性较高且能够靶向多种细胞,在临床前和临床研究中得到了较多的应用。不过在设计和生产的过程中,AAV载体有着诸多会影响其安全性和疗效的关键质量属性。生物质谱技术的发展和应用为生物大分子的研究提供了一个便捷的平台,尤其是在蛋白质序列、结构和相互作用方面。对于AAV载体而言,质谱技术可以实现衣壳蛋白比率、翻译后修饰、血清型、空衣壳比率的测定或表征,从而协助对AAV载体的质量控制。与现有方法相比,质谱技术具有样品需求量少、分析快速灵敏、适用于完整AAV载体的分析和质量分辨率高的优点,并且可以区分空衣壳、满衣壳和部分包封的衣壳。未来,通过将更加高效的蛋白质分离技术与质谱技术联用、开发新的信息处理软件平台和新的质谱检测方法,质谱技术有望在AAV载体的设计和生产中发挥更加重要的作用。