注射用疏血通的质量标准研究

目的制订注射用疏血通的质量标准。方法采用薄层色谱法鉴别水蛭;高效液相色谱法测定样品中次黄嘌呤的含量。结果次黄嘌呤在0.25~2.5μg呈良好的线性关系,r=0.999 7,加样回收率为98.4%,RSD=1.26%。结论该方法简便、准确、可以作为该制剂的质量控制标准。     

Uridine release during aminopyridine-induced epilepsy

Uridine, like adenosine, is released under sustained depolarization and it can inhibit hippocampal neuronal activity, suggesting that uridine may be released during seizures and can be involved in epileptic mechanisms. In an in vivo microdialysis study, we measured the extracellular changes of nucleoside and amino acid levels and recorded cortical EEG during 3-aminopyridine-induced epilepsy. Applying silver impregnation and immunohistochemistry, we examined the degree of hippocampal cell loss. We found that extracellular concentration of uridine, adenosine, inosine, and glutamate increased significantly, while glutamine level decreased during seizures. The release of uridine correlated with seizure activity. Systemic and local uridine application was ineffective. The number of parvalbumin- and calretinin-containing interneurons of dorsal hippocampi decreased. We conclude that uridine is released during epileptic activity, and suggest that as a neuromodulator, uridine may contribute to epilepsy-related neuronal activity changes, but uridine analogues having slower turnover would be needed for further investigation of physiological role of uridine.     

微生物来源的次黄嘌呤一磷酸盐脱氢酶抑制剂2264A和B的研究

利用自建的快速高效的次黄嘌呤一磷酸盐脱氢酶(Inosine monophosphate dehydrogenase,IMPDH)抑制剂的高通量筛选模型,筛选分离得到两个活性化合物2264 A和B,通过对其紫外、质谱、核磁等理化数据的分析确定2264 A为cyclopenol,2264 B为viridicatol。2264 A和B对IMPDH的IC50分别为69.68、11.86μmol/L;体外脾淋巴细胞增殖实验显示2264 A和B分别在322.6和65.8μmol/L浓度下可完全抑制由ConA活化的脾淋巴细胞增殖,表明2264 B在分子水平和细胞水平均表现出较好的免疫抑制活性,2264 A的活性相对较弱。     

头孢尼西钠与肌苷的配伍稳定性考察

目的考察注射用头孢尼西钠与肌苷注射液在0.9%氯化钠注射液中的配伍稳定性。方法在室温（[20±2）℃]条件下,观察8 h内配伍液的外观性状、pH值及紫外吸收光谱的变化,并用系数倍率法、紫外双波长等吸收法分别测定头孢尼西钠与肌苷的含量。结果两药在配伍后8 h内肌苷含量无明显变化、8 h时头孢尼西钠含量〈90%,其余外观性状、pH值及紫外吸收光谱等考察指标均无明显变化。结论注射用头孢尼西钠与肌苷注射液可在0.9%氯化钠注射液中配伍使用,但应在6 h内滴注完毕。     

复方肌苷口服液的制备及质量考察

目的:制备复方肌苷口服液并对其进行质量考察。方法:以肌苷为主药加入硒酸脂多糖制备口服液,采用高效液相色谱法测定含量,通过6个月加速试验和12个月长期留样试验考察其稳定性。结果:所制溶液为微黄色澄明溶液;肌苷检测浓度线性范围为10.24～35.84μg·mL-(1r=0.9999),平均回收率为100.2%,RSD=1.11%;稳定性试验中样品在考察期内各项指标均未见明显改变,pH值略有下降。结论:该制剂处方工艺合理可行,质量可控,稳定性良好。     

HPLC法同时测定康复新液中尿嘧啶、次黄嘌呤及肌苷含量

目的:建立HPLC法同时测定康复新液中的尿嘧啶、次黄嘌呤及肌苷含量.方法:色谱柱:Wates SunfireTM C18柱(250 mm×4.6 mm,5 μm );甲醇-3%甲醇溶液(含0.07%醋酸)梯度洗脱;检测波长254 nm;进样量:10 μl.结果:尿嘧啶在4.92～34.44 μg·ml-1 (r=0.999 9),次黄嘌呤在12.43～87.06 μg·ml-1(r=0.999 9),肌苷在24.77～173.40μg·ml-1(r=0.999 9)的浓度范围内呈良好的线性关系.尿嘧啶的平均回收率为102.63%(RSD=1.78%);次黄嘌呤的平均回收率为99.13%(RSD=1.89%);肌苷的平均回收率为98.98%(RSD=1.33%).结论:该法操作简便、准确,重复性、稳定性好,可用于同时检测康复新液中尿嘧啶、次黄嘌呤及肌苷的含量.     

Mycophenolic acid inhibits oleic acid-induced mesangial cell activation through both cellular reactive oxygen species and inosine monophosphate dehydrogenase 2 pathways

The synthesis of extracellular matrix (ECM) in mesangial cells (MCs) plays important roles in the development and progression of renal diseases, including chronic allograft nephropathy. Mycophenolic acid (MPA), an inhibitor of inosine monophosphate dehydrogenase 2 (IMPDH2), suppresses MC proliferation and ECM synthesis. However, the exact inhibitory mechanism of MPA on MCs has not been clearly elucidated. In this study we compared the inhibitory effects of MPA and IMPDH2 reduction [by using small interfering RNA (siRNA)] on oleic acid (OA)-induced fibronectin secretion and cellular reactive oxygen species (ROS) in mouse MCs. Growth-arrested MCs were stimulated with OA in the presence or absence of MPA, IMPDH2 siRNA, N-acetylcysteine (NAC), transforming growth factor beta (TGF-β) antibody or exogenous guanosine. Fibronectin secretion into the medium was examined by Western blot, dichlorodihydrofluorescein (DCF)-sensitive cellular ROS by fluorescence-activated cell scanning (FACS), TGF-β levels in the media by enzyme-linked immunosorbent assay (ELISA). OA increased fibronectin secretion, TGF-β and cellular ROS levels. A TGF-β neutralizing antibody effectively suppressed OA-induced fibronectin secretion. NAC and MPA completely suppressed OA-induced fibronectin secretion and decreased the levels of TGF-β and cellular ROS. However, IMPDH2 siRNA partly inhibited OA-induced MC activation. Exogenous guanosine successfully reversed the inhibitory effects of IMPDH2 siRNA on OA-induced MC activation. Pleiotropic inhibitory effect of MPA on OA-induced mouse MC activation was mediated via its antioxidant effect on cellular ROS production and partly via inhibition of IMPDH2 itself. Our results implicate ROS as an alternative therapeutic target for the prevention of hyperlipidemia-related glomerulopathy, chronic allograft nephropathy, and subsequent graft loss. Kyu Ha Huh and Hyung Joon Ahn contributed equally to this work and should be considered to be the co-first authors.     

肾移植受者TPMT和ITPA基因多态性与硫唑嘌呤所致不良反应关系的研究

目的探讨硫嘌呤甲基转移酶（TPMT）和三磷酸肌苷焦磷酸酶（ITPA）基因多态性与硫唑嘌呤（AZA）所致各种不良反应的关系。方法应用高效液相色谱法（HPLC）Ng定155例肾移植受者红细胞TPMT和ITPA活性,采用等位基因特异性的PCR（ASPCR）和限制性片断长度多态性（PCR-RFLP）的方法检测TPMT＊2、＊3A、＊3B和＊3C四种基因型,采用实时荧光定量PCR和PCR-RFLP的方法分别检测ITPA 94C〉A和IVS2＋21A〉C两种基因型,分析TPMT和ITPA活性和基因多态性与AZA所致不良反应的关系。结果155例肾移植受者中,120例患者未发生不良反应,将其作为对照组,平均TPMT活性为（37．26±11．60）U,平均ITPA活性为（97．9±34．7）U。另外35例患者由于发生了不良反应而停用AZA或减少了AZA的剂量。其中12例患者产生了血液毒性,平均TPMT活性为（22．92±10．67）U,明显低于对照组（P〈0．05）,其中仅2例患者为TPMT＊3C突变基因杂合子,AZA所致的血液毒性主要与患者红细胞TPMT活性低下有关。18例患者产生了肝脏毒性,其TPMT和ITPA活性离散度较大,其均值分别与对照组比较均无统计学差异（P〉0．05）。其中仅发现2例TPMT＊3C和4例ITPA94C〉A突变基因杂合子,TPMT和ITPA活性和基因多态性与AZA所致的肝脏毒性无明显相关性。5例患者产生了胃肠紊乱症状（包括1例患者同时产生了肝脏毒性）,其中有2例患者ITPA活性缺乏,为ITPA94C〉A突变基因纯合子,另外3例患者ITPA活性较低,为ITPA94C〉A突变基因杂合子。剩余1例患者出现了流感样症状,ITPA活性缺乏,为ITPA94C〉A突变基因纯合子。ITPA活性缺乏或ITPA94C〉A基因突变纯合子患者极易发生AZA所致胃肠紊乱症状和流感样症状的不良反应。结论一在服用AZA之前,对患者TPMT活性进行测定,有利于避免AZA所致血液毒性的发生。而对患者进行ITPA活性和基因多态性检测,有利于避免AZA所致的胃肠紊乱和流感样症状的产生,使AZA的使用更加安全、合理。     

手术修复严重肾损伤27例

目的报告２７例严重肾损伤的手术治疗体会，旨在降低伤肾切除率。方法术中采用静脉注射肌苷后，常温下阻断肾血流，根据肾破裂情况，分别采用肾部分切除、分层对位缝合、血管修补等修复手术。伤肾热缺血最长１０２分钟，最短２８分钟，平均５３分钟。结果２５例修复成功，２例肾切除，手术失肾率为７．４％；２５例在术后１４天～９个月行ＩＶＵ检查，伤（残）肾功能及形态良好；２１例随访４个月～１１年，血压、尿常规均正常。结论本法可明显降低严重肾损伤的肾切除率