癫痫是神经系统常见的慢性疾病,通常于儿童早期或青少年期发病,世界范围内的患病率约为0.76%[1] .截至2010年,全球癫痫患者总人数超过6 500万[2] ,男女比例无显著差异,但女性癫痫患者(WWE)中约有一半处于育龄期.癫痫及抗癫痫药物(AEDs)对胎儿有潜在的致畸和认知功能障碍等风险[3] ,并可能对孕妇造成创伤、早产、胎盘早剥、流产等不良影响[4] ,因此大多数育龄期WWE会选择各种避孕方式来防止意外妊娠.然而不幸的是,意外妊娠并不少见.一项研究显示,50%的育龄期WWE发生过意外妊娠,而口服避孕药避孕的失败是导致意外妊娠主要原因之一[5] .激素避孕药的避孕效果是高效的,但AEDs和激素避孕药之间存在着复杂的相互作用,这可能是避孕失败和／或癫痫控制不佳的原因.然而,很多临床医生缺乏对育龄期WWE避孕的充分认识,且育龄期WWE接受避孕咨询和管理的比例也很少.最近一项研究[6]发现,美国癫痫生育控制登记处调研了社区1 144例育龄期WWE的避孕实践,其中87.2%的患者曾就诊于神经科医生,但只有25.4%的患者接受了避孕方面的专业咨询.因此,本文将对激素避孕药与AEDs的相互作用、育龄期WWE避孕方式的选择进行概述,指出育龄期WWE避孕管理的方法和意义.

HECT, UBA and WWE domain-containing 1 (Huwe1), an E3 ubiquitin ligase involved in the ubiquitin-proteasome system, is widely ex-pressed in brain tissue. Huwe1 is involved in the turnover of numerous substrates, including p53, Mcl-1, Cdc6 and N-myc, thereby playing a critical role in apoptosis and neurogenesis. However, the role of Huwe1 in brain ischemia and reperfusion injury remains unclear. There-fore, in this study, we investigated the role of Huwe1 in an in vitro model of ischemia and reperfusion injury. At 3 days in vitro, primary cortical neurons were transduced with a control or shRNA-Huwe1 lentiviral vector to silence expression of Huwe1. At 7 days in vitro, the cells were exposed to oxygen-glucose deprivation for 3 hours and reperfusion for 24 hours. To examine the role of the c-Jun N-terminal kinase (JNK)/p38 pathway, cortical neurons were pretreated with a JNK inhibitor (SP600125) or a p38MAPK inhibitor (SB203508) for 30 minutes at 7 days in vitro, followed by ischemia and reperfusion. Neuronal apoptosis was assessed by TUNEL assay. Protein expression levels of JNK and p38MAPK and of apoptosis-related proteins (p53, Gadd45a, cleaved caspase-3, Bax and Bcl-2) were measured by western blot assay. Immunofluorescence labeling for cleaved caspase-3 was performed. We observed a significant increase in neuronal apoptosis and Huwe1 expression after ischemia and reperfusion. Treatment with the shRNA-Huwe1 lentiviral vector markedly decreased Huwe1 levels, and significantly decreased the number of TUNEL-positive cells after ischemia and reperfusion. The silencing vector also downreg-ulated the pro-apoptotic proteins Bax and cleaved caspase-3, and upregulated the anti-apoptotic proteins Gadd45a and Bcl-2. Silencing Huwe1 also significantly reduced p-JNK levels and increased p-p38 levels. Our findings show that downregulating Huwe1 affects the JNK and p38MAPK signaling pathways as well as the expression of apoptosis-related genes to provide neuroprotection during ischemia and reperfusion. All animal experiments and procedures were approved by the Animal Ethics Committee of Sichuan University, China in Janu-ary 2018 (approval No. 2018013).

目的 探讨汉族女性癫痫患者( women with epilepsy,WWE)性功能障碍与雌激素受体(estrogens receptor,ER)基因多态性的相关性.方法 连续收集2015年1月至2017年12月在扬州大学附属医院确诊的112例汉族已婚WWE,120例相匹配的健康汉族已婚女性作为对照组. WWE均接受抗癫痫药物(antiepileptic drugs,AEDs)治疗1年或以上.采用中文版女性性功能指数问卷( fe-male sexual function index,FSFI)调查研究对象的性功能;化学发光法测定血清泌乳素( prolactin, PRL)、卵泡刺激素、黄体生成素、雌二醇、孕酮和睾酮水平;聚合酶链反应-限制性片段长度多态性(polymerase chain reaction-restriction fragment length polymorphisms,PCR-RFLP)方法测定ER基因多态性.采用χ2 检验、独立样本t检验及二分类Logistic回归统计方法,比较两组研究对象性功能、性激素及ER基因型和等位基因频率的差异;分析两组研究对象性功能障碍与性激素及ER基因多态性的相关性.结果 (1)汉族 WWE发生性功能障碍的比例高达70. 5% ( 79/112),而对照组的比例是24. 2%(29/120). WWE 组血清 PRL 水平显著高于对照组[( 315. 40 ± 94. 66) mIU/L, ( 292. 09 ± 76. 65)mIU/L],差异有统计学意义(t=2. 072,P=0. 039). (2) WWE组ER基因PvuⅡ C、AluⅠA等位基因频率显著高于对照组,差异有统计学意义(P=0. 001;P=0. 001). (3)二元Logistic回归分析发现PvuⅡ CC基因型和血清睾酮水平与 WWE 性功能障碍独立相关(OR=6. 074,95% CI:1. 257~29. 352,P=0. 025;OR=0. 412,95%CI:0. 201~0. 842,P=0. 015).结论 ERα基因PvuⅡ多态性可能与汉族WWE性功能障碍的易感性有关,PvuⅡ CC基因型可能是WWE性功能障碍的风险基因型.

酒糟是良好的沼气发酵原料,但其发酵中间产物极易累积导致体系酸化.采用批次试验研究中温条件下酱酒丢糟沼气发酵特性.以总固体(TS)含率分别为4.3％和7.8％的酱香白酒丢糟为发酵原料,在发酵30 d和50 d时,甲烷产量分别为287 mL/g TS和360 mL/g TS;TS去除率分别为20.8％和27.4％.酱酒糟发酵过程中主要的中间代谢产物有乙酸、丙酸和丁酸等挥发性有机酸;此外,还检测出芳香族化合物有苯酚、对甲酚、苯甲酸和苯丙酸;其中丙酸在酒糟降解过程中明显累积(2894-4495 mg/L),推测丙酸累积为影响反应器稳定运行的重要因素.利用高通量测序技术对两组中温厌氧酒糟驯化培养系JO和JN进行解析.细菌结果表明,拟杆菌门(Bacteroidetes)、厚壁菌门(Firmicutes)、互养菌门(Synergistetes)和阴沟单胞菌门"Cloacimonetes"(WWE1)为主要的优势菌群.WWE1中未培养W5属和W27属在酒糟厌氧消化系统中占比为13.8％-30.67％.古菌解析检测到乙酸营养型产甲烷菌Methanosarcina mazei及氢营养型产甲烷菌Methanofollis ethanolicus近缘微生物为优势古菌.本研究表明丙酸累积是影响酱酒丢糟沼气发酵稳定性的重要因素,WWE1中未培养微生物在酒糟厌氧发酵体系中发挥重要作用.

泛素-蛋白酶体系统在蛋白质降解时发挥着重要的作用.泛素化过程需要E1泛素激活酶、E2泛素结合酶、E3泛素连接酶协同完成.本研究组前期研究证明E3泛素连接酶HUWE1(HECT,UBA and WWE domain containing 1)可降解表皮生长因子受体(epidermal growth factor receptor,EGFR),抑制肾小管间质纤维化.为了进一步明确HUWE1抑制肾小管间质纤维化的机制,本研究鉴定了参与HUWE1降解EGFR过程的E2泛素结合酶.通过real-time PCR观察可能与HUWE1发生相互作用的候选E2泛素结合酶在肾脏损伤模型中的表达变化.用小鼠单侧输尿管结扎(unilateral ureteral obstruction,UUO)模型的肾组织和转化生长因子-β(transforming growth factor-β,TGF-β)刺激的人肾脏近端小管上皮细胞(HK-2细胞)来检测候选E2泛素结合酶的表达量变化.结果显示,与对照组相比,E2泛素结合酶UBE2Q2在UUO术后小鼠肾脏组织中mRNA与蛋白水平均显著下调,在TGF-β刺激的HK-2细胞中UBE2Q2 mRNA和蛋白表达水平也显著下调,与HUWE1表达变化趋势一致,表明在肾脏损伤时E2泛素结合酶UBE2Q2表达水平与HUWE1具有协同性.免疫共沉淀(co-immunoprecipitation,Co-IP)和细胞免疫荧光染色结果验证了HUWE1与UBE2Q2的相互作用.在HK-2细胞中敲低UBE2Q2后,HUWE1、EGFR与泛素的结合均显著降低.上述结果提示,UBE2Q2可能是与HUWE1相互作用的E2泛素结合酶,参与HUWE1对肾小管间质纤维化的抑制作用.