CBP/P300介导的组蛋白H3K9乙酰化修饰对GATA4表达的影响

目的 明确GATA4在小鼠胚胎心脏发育过程中的时序表达,并阐明其组蛋白修饰调控机制.方法 选取胎龄11.5 d(E11.5)至新生0.5d胎鼠心脏,RT-PCR检测胎鼠心肌细胞中GATA4mRNA表达水平,CHIP-QPCR检测GATA4启动子区组蛋白H3K9ac水平及与CBP/P300的结合水平.用CBP30干预心肌祖细胞,RT-PCR检测心肌祖细胞在不同浓度CBP30(0.5、1、2、4μmol/L)干预不同时间点(6、12、24、36 h)后GATA4 mRNA表达水平,CHIP-QPCR检测心肌祖细胞中GATA4启动子区组蛋白H3 K9 ac水平及与CBP/P300的结合水平.结果 ①小鼠胚胎心脏发育过程中GATA4表达量,E14.5明显高于E11.5(P ＜0.05);CHIP结果显示E14.5 GATA4启动子区组蛋白H3 K9 ac水平高于E11.5(P ＜0.05);E14.5与CBP/P300的结合水平亦高于E11.5(P ＜0.05).②CBP30干预心肌祖细胞后,GATA4表达量较对照组明显降低(P＜0.05).CHIP结果显示CBP30组GATA4启动子区组蛋白H3K9ac水平及与CBP/P300的结合水平较对照组显著降低(P＜0.05).结论 CBP/P300可通过介导组蛋白H3 K9乙酰化修饰参与调控GATA4在小鼠胚胎心脏发育过程中的时序表达.     

柱前衍生超高效液相色谱法分析太子参多糖中单糖的组成

目的  建立柱前衍生超高效液相色谱（UPLC）法,测定太子参多糖中单糖的组成。方法  采用水提醇沉法提取太子参多糖,2 mol/L硫酸水解后加入1-苯基-3-甲基-5-吡唑啉酮进行衍生化,采用UPLC法测定太子参多糖中单糖的衍生物。采用沃特世C18超高效液相色谱柱（100.0 mm×2.1 mm,1.7μm）色谱柱,以乙腈为流动相A,0.1 mol/L磷酸盐（pH 6.8）缓冲液为流动相B,梯度洗脱,检测波长250 nm。通过聚类分析和主成分分析,对不同产地太子参单糖进行质量评价。结果  太子参多糖由半乳糖、D-甘露糖、鼠李糖、阿拉伯糖、D-无水葡萄糖、D-葡萄糖醛酸、D-半乳糖醛酸7种单糖组成,不同产地太子参多糖中单糖组成稍有差别,通过主成分分析和聚类分析评价,10个不同产地太子参单糖聚成3类,并按照单糖含量对各产地进行排名。结论  柱前衍生UPLC法表明,太子参多糖主要由半乳糖醛酸、半乳糖和阿拉伯糖构成,并且贵州施秉县地区产太子参单糖含量较高。

     

清金化痰汤对慢性阻塞性肺疾病急性加重期模型大鼠肺组织 Foxp3和 RORγt 表达的影响

目的探讨清金化痰汤对慢性阻塞性肺疾病急性加重期(AECOPD)模型大鼠肺组织叉头翼状螺旋转录因子P3(Foxp3)、维甲酸相关孤核受体γt(RORγt)表达的影响。方法 40只SPF Wistar雄性大鼠随机分为正常组、模型组、清金化痰汤组、克拉霉素组。除正常组外,其余组采取气道滴注脂多糖(LPS)联合烟熏的方法建立AECOPD气道黏液高分泌模型,并同时连续30 d分别给予生理盐水、清金化痰汤(8.4 g/kg)、克拉霉素(50 mg/kg)灌胃。实验第31天,提取大鼠肺组织,制作病理切片,并采用免疫组化法检测肺组织中黏蛋白5AC(Muc5AC)、中性粒细胞弹性蛋白酶(NE)的表达,观察Foxp3和RORγt的表达,免疫印迹法检测肺组织中Foxp3和RORγt的蛋白表达。结果与正常组比较,模型组Foxp3表达量无明显差异(P0.05),RORγt、Muc5AC、NE表达显著升高,Foxp3/RORγt显著下降(P0.01);清金化痰汤组与模型组比较,Foxp3表达量无明显差异(P0.05),RORγt、Muc5AC、NE表达显著降低,Foxp3/RORγt显著升高(P0.01或P0.05)。清金化痰汤组与克拉霉素组比较,Muc5AC显著降低(P0.05)。结论清金化痰汤可显著下调模型大鼠肺组织中RORγt蛋白表达,上调Foxp3/RORγt,这可能是清金化痰汤调节AECOPD黏液高分泌的机制之一。     

急性心肌梗死恢复期V1导联P波终末电势与左室舒张功能的相关性研究

目的  探讨急性心肌梗死（AMI）患者恢复期心电图V1导联P波终末电势（Ptfv1）与左心室舒张功能的相关性。方法  选择78例AMI后6周左右[（41.76±4.28）d]心电图Ptfv1≤-40 mm/s的患者为观察组,以及同期86例心电图Ptfv1>-40 mm/s的AMI患者为对照组。比较两组间超声心动图左房内径（LAD）、左室舒张期直径（LVDd）、左室等容舒张时间（LVRT）、二尖瓣舒张早期流速峰值（E）、二尖瓣舒张晚期流速峰值（A）、左心室射血分数（LVEF）、组织多普勒舒张早期二尖瓣环水平心肌速率峰值（E''）、E/A及E/E''等。结果  观察组LVDd、LAD、LVRT增大（P <0.05）,观察组LVEF减低（P <0.01）,观察组中E/A值降低（P <0.025）,观察组E及E''峰值减小（P <0.05）,E/E''值増高（P <0.05）,观察组E''<9 cm/s、E/E''>8及15的例数增多（P <0.05）。结论  在AMI恢复期患者中,Ptfv1值异常反映左心室舒张功能减退。测量Ptfv1是简单易行的初步评估左室舒张功能方法。

     

胃癌组织中Ki-67、P53、EGFR的表达及其与TNM分期的相关性研究

【摘要】目的：探究分析胃癌组织中Ki-67、P53、EGFR的表达及其与TNM分期的相关性。方法：随机选取我院2014年1月～2016年1月收治的120例胃癌患者及其手术切除的病理标本,收集患者的临床治疗资料,同时电话随访相关病例。运用免疫组化MaxVisionTM2/HRP染色法对胃癌组织中Ki-67、P53、EGFR抗原相关基因蛋白产物进行检测,对其表达水平与临床TNM分期的相关性及其临床意义进行分析,最后统计分析所有结果。结果：胃癌中P53、Ki-67、EGFR蛋白阳性表达率分别为26.67%（32/120）、40.00%（48/120）、12.50%（15/120）;高分化腺癌、有局部淋巴结转移、TNMⅡ期+Ⅲ期+Ⅳ期患者的P53、Ki-67联合检测阳性表达率均显著高于中低分化腺癌、粘液腺癌及印戒细胞癌、无局部淋巴结转移、TNMⅠ期患者（P＜0.05）。结论：胃癌患者血清P53、Ki-67、EGFR水平表达与TNM分期具有密切关系,三者联合检测在患者预后判断及评估中具有显著的指导价值,为临床上分子靶向治疗的研究提供了借鉴依据。     

Increased Ca2+ channel currents in cerebellar Purkinje cells of the ataxic groggy rat

The ataxic groggy rat (strain name; GRY) is an autosomal recessive neurological mutant found in a closed colony of Slc:Wistar rats. Recent genetic analysis has identified the missense (M251K) mutation in the alpha(1) subunit of the Ca(V)2.1 (P/Q-type) voltage-dependent Ca(2+) channel gene (Cacna1a) of GRY rat. In this study, we found that high-voltage-activated (HVA) Ca(2+) channel currents in acutely dissociated Purkinje cells of GRY rats showed increased (not decreased) current density and depolarizing shift of the activation and inactivation curves compared with those of normal Wistar rats. In contrast low-voltage-activated (LVA) Ca(2+) channel currents of GRY rats showed no significant changes. These results suggest that functional alteration of Ca(2+) channel currents in cerebellar Purkinje cells of GRY rats is attributed to the change of HVA Ca(2+) channel currents, and that increased HVA Ca(2+) channel function underlies the cerebellar dysfunction and ataxic phenotype of GRY rats.     

Affective visual event-related potentials: arousal, repetition, and time-on-task

Affective stimulus pictures that differed in valence (unpleasant, neutral, and pleasant) were repeated as targets in an oddball task to elicit event-related potentials (ERPs) in young female adults. Each picture target was repeated consecutively four times, with picture order counterbalanced and time-on-task influences assessed across subjects. Response time decreased from the first to second stimulus presentation and remained stable. Stimulus repetition was associated with voltage increases for N1, P2, N2, and P3, from initial to subsequent presentations. Arousal effects did not interact with stimulus repetition at any latency range. Time-on-task was associated with decreased voltages for the N2 and P3 potentials but was unaffected by stimulus valence. The findings suggest affective arousal, stimulus repetition, and time-on-task independently modulate ERP outcomes at overlapping time ranges. Theoretical implications are discussed.     

Fructosamine-6-phosphates are deglycated by phosphorylation to fructosamine-3,6-bisphosphates catalyzed by fructosamine-3-kinase (FN3K) and/or fructosamine-3-kinase-related-protein (FN3KRP)

Nonenzymatic glycation of proteins and some phospholipids by glucose and other reducing sugars (a.k.a Maillard reaction) is an unavoidable result of the coexistence of these sugars and the affected macromolecules in living systems. The consequences of this process are deleterious both in the intracellular and extracellular environments as evidenced by the close association between increased nonenzymatic glycation and complications of diabetes. Because of these considerations, we have proposed that the intrinsic toxicity of glucose and other sugars is counteracted in vivo by active deglycation mechanisms including transglycation of Schiff's bases and FN3K-dependent breakdown of fructosamines. While this modified hypothesis is receiving increasing experimental support, several issues regarding glycation/deglycation remain unresolved. Two such important questions are In this paper we propose a resolution of both these quandaries by proposing that fructosamine-6-phosphates are deglycated by phosphorylation to fructosamine-3,6-bisphosphates catalyzed by FN3KRP and/or possibly FN3K. We provide some preliminary evidence in support of this hypothesis and outline experimental approaches for definitive tests of this hypothesis. The potential medical implications of this finding are not clear yet but, if correct, this observation is likely to have a major impact on our understanding of the very basic and hitherto unexplored aspect of glucose metabolism and chemistry in vivo. One can imagine that, at some point in the future, measurement of FN3K/FN3KRP activity may be of diagnostic value in assessing an individual's susceptibility to diabetic complications. Further down the road, one can also envision a gene therapeutic intervention to bolster FN3K/FN3KRP-based antiglycation defenses.     

Brain regions and genes affecting postural control

Postural control is integrated in all facets of motor commands. The role of cortico-subcortical pathways underlying postural control, including cerebellum and its afferents (climbing, mossy, and noradrenergic fibers), basal ganglia, motor thalamus, and parieto-frontal neocortex has been identified in animal models, notably through the brain lesion technique in rats and in mice with spontaneous and induced mutations. These studies are complemented by analyses of the factors underlying postural deficiencies in patients with cerebellar atrophy. With the gene deletion technique in mice, specific genes expressed in cerebellum encoding glutamate receptors (Grid2 and Grm1) and other molecules (Prkcc, Cntn6, Klf9, Syt4, and En2) have also been shown to affect postural control. In addition, transgenic mouse models of the synucleinopathies and of Huntington's disease cause deficiencies of motor coordination resembling those of patients with basal ganglia damage.