左旋多巴诱发异动症大鼠纹状体前强啡肽原基因与DARPP-32蛋白磷酸化的关系

目的研究左旋多巴诱发异动症(levodopa-induced dyskinesias,LID)大鼠纹状体内前强啡肽原(prodynorphin,PDyn)基因表达与DARPP-32蛋白磷酸化状态的变化,探讨LID时直接通路过度活化的机制。方法6-羟多巴胺(6-OHDA)大鼠模型应用左旋多巴治疗28 d诱发LID大鼠模型。采用原位杂交技术检测PDyn mRNA水平,逆转录-聚合酶链反应(RT-PCR)及免疫印迹技术检测LID大鼠纹状体内总DARPP-32的mRNA与蛋白表达及其Thr-34位点磷酸化水平。结果LID大鼠毁损侧PDyn mRNA表达(0.3662±0.0625)较对照组(0.2085±0.0573)及L-dopa治疗组(0.2235±0.0582)明显增高,差异有统计学意义(P<0.01)。LID大鼠毁损侧Thr-34位点磷酸化的DARPP-32蛋白水平(1075.2±103.3)较对照组(198.7±49.5)及L-dopa治疗组(213.9±58.9)明显增高,差异均有统计学意义(P<0.01)。结论DARPP-32蛋白的Thr-34位点的磷酸化水平的改变是LID时多巴胺D1受体介导的直接通路异常活化的关键因素之一。     

肺结核抗结核药物致肝损伤患者HMGB1基因多态性

目的探讨肺结核抗结核药物致肝损伤患者HMGB1基因多态性。方法选择2016年8月-2019年8月中南大学湘雅医院附属海口医院滨江分院收治的肺结核患者200例,发展成ATDH的患者纳入ATDH组(n=79),未发展成ATDH的患者纳入非ATDH组(n=121),检测HMGB1基因单核苷酸多态性(SNPs)位点多态性,采用Logistic回归分析HMGB1多态性位点与ATDH的关系。结果ATDH组HMGB1基因rs1045411位点GG、GA、AA基因型分布频率和G、A等位基因频率与非ATDH组比较,差异有统计学意义(P<0.05),rs1412125、rs1360485以及rs2249825位点基因型分布和等位基因频率与非ATDH组比较,均无统计学差异;在共显性、显性、隐性模式下,ATDH组HMGB1基因rs1045411位点基因型分布与非ATDH组比较,差异有统计学意义(P<0.05),与GG基因型比较,GA和AA基因型与肺结核易感性有关,在超显性模式下,ATDH组HMGB1基因rs1045411位点基因型分布与非ATDH组比较,无统计学差异,而在超显性、共显性、显性以及隐性模式下,rs1412125、rs1360485以及rs2249825位点基因型分布与非ATDH组比较,均无统计学差异;rs1045411位点基因多态性与ATDH的发生存在相关性,且突变型诱发ATDH可能性更大,而rs1412125、rs1360485、rs2249825位点基因多态性与ATDH发生风险无关。结论HMGB1基因为ATDH的易感基因,其rs1045411位点多态性与ATDH的发生密切相关,其中突变型发生ATDH风险升高。     

Critically ill,tubular injury,delayed early recovery: characteristics of acute kidney disease with renal oxalosis

ObjectsThis study aimed to analyze the clinicopathological features of acute kidney disease (AKD) with renal oxalosis.MethodsData for biopsy-proven AKD with oxalosis diagnosed from Jan 2011 to Oct 2018 was collected. The underlying diseases, dietary habits, clinical and pathological characteristics of newly emerging kidney disease were analyzed. The long-term renal prognosis was observed.ResultsA total of 23 patients were included, comprised of 18 men and 5 women with a mean age of 51.6 ± 15.9 years. The peak Scr was 669.9 ± 299.8 μmol/L, and 95.7% of patients had stage 3 acute kidney injury (AKI). Drug-induced tubulointerstitial nephritis (TIN) was the most common cause (65.2%) of AKD, followed by severe nephrotic syndrome (17.4%). All patients had pathological changes indicating TIN, and 11 patients were complicated with the newly emerging glomerular disease (GD). The risk of oxalosis caused by increased enterogenous oxalate absorption accounted for only 26.1%, and others came from new kidney diseases. The majority (75%) of abundant (medium to severe) oxalosis occurred in patients without GD. There were no significant differences in the score for tubular injury (T-IS) and interstitial inflammation with different severities of oxalosis. Rate of Scr decrease (ΔScr%) at 2 weeks was negatively correlated with the severity of oxalosis (R = −0.542, p = 0.037), score for T-IS (R = −0.553, p = 0.033), and age (R = −0.736, p = 0.002). The decrease in Scr at 4 weeks was correlated with T-IS (R = −0.433), but had no correlation with oxalosis.ConclusionsThe present findings revealed that 95.7% of AKD with secondary renal oxalosis occurred in critically ill patients. AKD from tubular injury was the prominent cause. Severe oxalosis contributed to delayed early recovery of AKD.     

中药抗药源性肝损伤评价模型及作用机制研究进展

肝脏是药物在机体内代谢的主要场所。药源性肝损伤（drug-induced liver injury,DILI）是药物过量或不当使用导致的机体不良反应之一。非甾体类抗炎药、抗菌药等均可能引起DILI。DILI发生机制主要与细胞色素P450酶、线粒体功能障碍、药物代谢反应、氧化应激、炎症反应等密切相关。中药可通过抑制氧化应激、炎症反应、减少细胞凋亡与影响细胞色素P450酶系统等发挥治疗肝损伤作用。通过对DILI发生机制、动物模型建立方法及中药干预机制等方面进行总结归纳,为DILI动物模型的合理选择及中药防治机制研究提供参考。     

Detection of drug-induced apoptosis by flow cytometry after alkaline extraction of ethanol fixed cells

A new flow cytometric method was developed to detect apoptotic cells with fragmented DNA and to determine cell cycle distribution of viable cells, in the same sample, by propidium iodide staining. Apoptosis, in HT58 human B lymphoma cells, was induced by etoposide and/or by staurosporine. Using appropriate alkaline solutions (between 1-10 mN NaOH in 150 mM saline) followed by neutralization with buffer solution, the fragmented DNA can be extracted quantitatively from ethanol fixed cells. Further, good resolution of the cell cycle distribution can be obtained in unimpaired cells without RNase treatment. Furthermore, unlike the widely used hypotonic-detergent extraction of unfixed cells, the suggested extraction method can prevent drug-induced disintegration of dead cells when karyorrhexis occurs. This work was supported by Hungarian National Research Foundation (OTKA I/352 and OTKA II/2622).     

利用医院集中监测药物不良反应管理系统适时监测药物性肝损害

目的 :研究利用医院集中监测药物不良反应管理系统监测药物性肝损害的方法。方法 :应用自制程序 ,从医院住院患者数据库中提取2001年12月～2002年2月丙氨酸氨基转移酶、门冬氨酸氨基转移酶和总胆红素异常的住院患者资料 ,并进行回顾性与适时性分析。结果 :可能涉及药物不良反应的有50例 ,相关药物有11类、30种 ;属不合理用药的有11例 ,相关药物有10种。结论 :通过这种方法可及时获得药物性肝损害发生的信息 ,同时这也为医院集中开展药物不良反应监测提供了一种新的思路。     

肠源性内毒素血症对慢性实验性肝损伤的影响及防治对策的研究

目的探讨肠源性内毒素血症对慢性实验性肝损伤的影响及防治对策。方法将Wistar大鼠随机分为3组:正常对照组、慢性肝损伤组及双利肝治疗组(双利肝组),每组10只。采用CCl4复合因素复制慢性肝损伤模型,4周末处死动物,取血检测血浆内毒素(ET),血清丙氨酸氨基转移酶(ALT)、γ-干扰素(IFN-γ)、白介素-2(IL-2)和白介素-4(IL-4),同时观察肝脏组织学变化。结果①CCl4所致的慢性肝损伤组与正常大鼠组相比,血浆ET、血清ALTI、L-4水平明显增高(P<0.05),血清IL-2、IFN-γ水平明显降低(P<0.05)。②与CCl4所致慢性肝损伤组相比,双利肝组血浆ET、血清ALTI、L-4水平明显下降(P<0.05),血清IL-2、IFN-γ水平明显增高(P<0.05)。③组织学观察:经双利肝治疗后肝脏组织学明显改善。结论CCl4所致慢性肝损伤大鼠存在肠源性内毒素血症,后者可能导致细胞免疫功能紊乱。经双利肝治疗后血浆内毒素水平明显下降,Th1/Th2比例得到调整,肝细胞损伤减轻。