耐力运动对关节炎大鼠氧化应激与软骨炎性表达及CHRNA7信号的影响

目的 探讨耐力运动对关节炎大鼠氧化应激、软骨炎性表达及CHRNA7信号表达的影响。方法 将50只SD大鼠随机分为健康组、关节炎组、低耐力组、中耐力组、高耐力组，每组10只。检测各组大鼠行为学及氧化应激水平差异性；采用热板致痛阈值（PWTL）及机械性撤足阈值（PWT）；酶联免疫法检测各组大鼠炎性因子水平；采用Western Blot法检测滑膜组织中a7nAChR、STAT3表达情况；HE染色观察大鼠膝关节软骨组织的形态。结果 HE染色结果显示，健康组大鼠膝关节软骨表面平整光滑，无裂缝或缺损；关节炎组大鼠膝关节软骨的表面不平整，有缺损；低耐力组大鼠膝关节软骨的表面比较光滑、平整；中、高耐力组大鼠膝关节软骨的表面平整，软骨细胞排列较为整齐。 与健康组相比，关节炎组TNF-α、IL-1水平升高（P＜0.05）；与关节炎组相比，低耐力组、中耐力组和高耐力组TNF-α、IL-1水平降低明显（P＜0.05）；与低耐力相比，中耐力组TNF-α、IL-1降低明显（P＜0.05）；与中耐力组相比，高耐力组TNF-α、IL-1水平升高（P＜0.05）。与健康组相比，关节炎组大鼠PWTL、PWT、SOD、GSH-Px、α7nAChR、STAT3表达水平明显降低，MDA水平升高(P＜0.05)；与关节炎组大鼠相比，低、中和高耐力组大鼠PWTL、PWT、SOD、GSH-Px、α7nAChR、STAT3水平升高，MDA水平降低(P＜0.05)；与低耐力组相比，中耐力组PWTL、PWT、SOD、GSH-Px、α7nAChR、STAT3水平升高，MDA水平降低(P＜0.05)；与中耐力组相比，高耐力组PWTL、PWT、SOD、GSH-Px、α7nAChR、STAT3水平下降，MDA水平升高(P＜0.05)。结论 中强度耐力运动可抑制TNF-α、IL-1炎性水平，改善疼痛水平及氧化应激反应，对关节起到保护作用，其机制可能与调控α7nAChR/STAT3蛋白表达有关。     

菊花脑芳樟醇合酶基因CnTPS1的克隆与原核表达分析＊

目的 克隆菊花脑芳樟醇合酶基因CnTPS1的全长编码序列，利用原核系统表达融合蛋白，为进一步研究该基因在菊花脑萜类合成中的功能提供理论依据。方法 以菊花脑基因组数据为基础，设计特异性引物，PCR扩增CnTPS1的全长编码序列，利用生物信息学分析软件分析序列特征。利用qRT-PCR技术分析CnTPS1基因在不同组织中的基因表达量。构建原核表达载体，体外诱导目的蛋白表达。结果 CnTPS1编码序列全长1749bp，编码582个氨基酸；基因表达模式分析表明该基因在茎和管状花中表达量较高；原核表达系统能诱导出67.58kDa大小的蛋白。结论 首次从菊花脑中克隆得到一个芳樟醇合酶基因CnTPS1，运用生物信息学方法对其编码蛋白的理化性质、结构特征等进行了分析预测，分析了该基因的组织表达模式，并在原核表达系统中成功诱导表达出目标蛋白，这些结果将为菊花脑萜类合酶基因的功能以及萜类物质生物合成途径的解析提供理论依据。     

晚期EGFR突变肺腺癌Ki-67表达与EGFR-TKIs治疗疗效的相关性北大核心

目的:分析晚期EGFR突变型肺腺癌(LADC)组织中Ki-67表达和其他临床病理特征与EGFR-TKIs客观疗效和生存预后的相关性。方法:回顾性分析2017年05月至2020年05月我院102例晚期EGFR突变型LADC中Ki-67表达情况与其他临床特征,所有患者均接受第一代EGFR-TKIs 6周后进行客观疗效评价,无进展生存期(progression-free survival,PFS)为主要研究终点。结果:Ki-67表达可较为准确地预测EGFR-TKIs的客观缓解率(object response rate,ORR)[AUC=0.7690(0.6705~0.8675),P<0.0001],Ki-67表达与EGFR-TKIs客观疗效呈负性相关(P=0.000),分期(P=0.170)、EGFR-TKIs(P=0.112)、ECOG PS(P=0.551)、EGFR突变(P=0.163)与ORR无明显相关。Ki-67高表达相对低表达,可显著缩短PFS(P<0.0001);单因素分析发现ECOG PS(HR 1.740,P=0.001)、EGFR突变(HR 1.656,P=0.023)、分期(HR 1.487,P=0.005)、Ki-67表达(HR 1.027,P=0.000)为影响PFS的相关因素;多因素分析发现,Ki-67表达(HR 1.040,P=0.000)为影响PFS的独立预后因素。结论:晚期EGFR突变型LADC组织中Ki-67高表达可降低EGFR-TKIs的ORR,缩短PFS,Ki-67表达在晚期EGFR突变LADC中的意义值得进一步探索。     

右归饮对去卵巢血管钙化大鼠NMDAR1蛋白表达的影响

目的:观察右归饮对去卵巢血管钙化中谷氨酸受体变化的影响。方法:将40只远交群雌性大鼠(sprague dawley,SD)随机分为假手术组、模型组、低剂量组、高剂量组和辛伐他汀组,采用皮下注射华法令加维生素D3制备血管钙化模型,造模后,低剂量组、高剂量组分别给予右归饮浓缩液5.35 g/(kg·d)和16.05 g/(kg·d)灌胃,辛伐他汀组给予辛伐他汀混悬水溶液5.25 mg/(kg·d)灌胃,假手术组和模型组给予等量生理盐水,共12周;实验结束时采集血浆标本测定雌二醇及血脂四项,采用苏木精-伊红染色法(HE染色)观察动脉钙化的情况,并采用免疫印迹试验检测N-甲基-D-天门冬氨酸受体(NMDAR,NR)1蛋白的表达。结果 :模型组大鼠雌二醇、甘油三酯含量和NR1蛋白表达降低,P<0.05,低密度脂蛋白升高,P<0.05;高剂量组和低剂量组雌二醇、甘油三酯含量升高,NR1蛋白表达增加,P<0.05;高剂量组、低剂量组和辛伐他汀组低密度脂蛋白的含量降低,P<0.05;辛伐他汀组雌二醇无变化,P>0.05。结论:右归饮可抑制大鼠去卵巢血管钙化的形成,可能与提高雌激素水平和NR1蛋白的表达有关。     

微小RNA在人类压疮组织中的异常表达

目的建立人类压疮组织微小RNA表达谱。方法收集本院24例压疮组织临床样本,选取其中4例用于压疮微小RNA芯片制备。采用生物信息学算法比较压疮组织与正常组织中差异表达的微小RNA,获得压疮微小RNA表达谱。进一步利用20例测试样本,采用实时荧光定量-逆转录-聚合酶链反应对所鉴定的表达谱进行验证。结果获得了12个差异表达的微小RNA,其中包括5个表达水平上调和7个表达水平下调的微小RNA,其中表达水平下调的miR-17差异表达水平最显著。结论作者建立了一组由12个微小RNA组成的压疮组织微小RNA表达谱,为压疮发生及发展分子调控机制研究提供了丰富的素材。     

A RGG motif protein is involved in Toxoplasma gondii stress-mediated response

The molecular mechanisms controlling gene expression are still poorly understood in apicomplexan parasites. Here, we report the characterization of a homolog of the single strand binding proteins (named TgSsossB) in Toxoplasma gondii. We previously showed that TgSsossB interacts with the TgAlba proteins that are involved in translation regulation. We examined the role of TgSsossB in stress-mediated response, and particularly the role of its arginine–glycine–glycine (RGG) repeats domain. TgSsossB recombinant protein is able to bind to single strand DNA and RNA in a sequence-independent manner, but not to double stranded DNA. We showed that the RGG motif is not involved in this ability to bind to nucleic acid. We produced a mutant tagged strain lacking the RGG motif of TgSsossB using the knock-in strategy. We observed that this strain exhibited a fitness defect compared with the parental parasites. Moreover, the mutant strain produced fewer plaques in stress conditions, a defect that is due to a slow growth phenotype when extracellular parasites are exposed to stress. At the molecular level, we showed that the TgSsossB protein lacking a RGG motif lost its ability to interact with TgAlba2 and an isoform of TgAlba1, indicating that the TgAlba complex is likely non-functional in those parasites. Thus, our findings define the RGG domain of TgSsossB as a protein–protein interaction platform and underline the role of the TgAlba–TgSsossB complex in stress-mediated response.     

The possible antianginal effect of allopurinol in vasopressin-induced ischemic model in rats

The anti-anginal effects of allopurinol were assessed in experimental model rats of angina and their effects were evaluated with amlodipine. In the vasopressin-induced angina model, oral administration of allopurinol in dose of 10 mg/kg revealed remarkably analogous effects in comparison with amlodipine such as dose-dependent suppression of vasopressin-triggered time, duration and severity of ST depression. In addition, allopurinol produced dose dependent suppression of plasma Malondialdehyde (MDA) level, systolic blood pressure, cardiac contractility and cardiac oxygen consumption; while in contrast, amlodipine minimally suppressed the elevation of plasma MDA level. Endothelial NO synthase (eNOS) expression, serum nitrate were strikingly increased, however lipid profile was significantly reduced. Seemingly, allopurinol was found to be more potent than amlodipine – a calcium channel antagonist. To conclude, it was explicitly observed and verified that on the ischemic electrocardiography (ECG) changes in angina pectoris model in rats, allopurinol exerts a significant protective effects, reminiscent of enhancement of vascular oxidative stress, function of endothelial cells, improved coronary blood flow in addition to the potential enhancement in myocardial stress. Moreover, our findings were in conformity with several human studies.