Clonidine-induced locomotor hyperactivity in rats

Summary The -adrenergic agonist, clonidine, causes sedation in normal rats. The present study demonstrates that clonidine evokes strong locomotor stimulation in rats pretreated with 6-hydroxydopamine plus reserpine. Similar, but less intensive hyperactivity is observed in rats given clonidine after combined pretreatment with 6-hydroxydopamine plus p-chlorophenylalanine plus -methyl-p-tyrosine, or with reserpine plus low doses of yohimbine. The -adrenolytic drugs, phenoxybenzamine, phentolamine and aceperone, as well as high doses of yohimbine, antagonise the clonidine-induced locomotor stimulation; in contrast, the dopamine receptor blocking agents, pimozide and spiroperidol, exert no antagonistic effect. The results indicate that in the brain of normal animals, clonidine predominantly activates presynaptic -adrenoceptors on noradrenergic neurones and thereby induces sedation. After destruction of the noradrenergic fibres by 6-hydroxydopamine plus reserpine, activation of postsynaptic -adrenoceptors prevails so that hyperactivity results.This study was supported by Polish Academy of Sciences (10.4). Preliminary accounts were presented at the Pharmacology Meeting, Hannover, September 14–17, 1976 and at the 1 st Joint Symposium of Hungarian and Polish Pharmacological Societies, Zakopane, October, 13–15, 1976     

长链非编码RNA DLX6-AS1 与微小RNA-346在糖尿病足72 例血清中的表达及其临床意义

目的探讨长链非编码RNA(lncRNA)DLX6-AS1与微小RNA(miR)-346在糖尿病足病人血清中的表达及其临床意义。方法收集2017年5月至2018年12月南阳市中心医院收治的72例糖尿病足病人为糖尿病足组,选取同期该院收治的60例2型糖尿病无糖尿病足病人为糖尿病组,取同期于该院进行健康体检的健康志愿者55例作为对照组。实时荧光定量逆转录聚合酶链反应(qRT-PCR)检测各组受检者血清中DLX6-AS1、miR-346的表达水平;采用酶联免疫吸附测定(ELISA)检测血管细胞黏附分子-1(VCAM-1)、成纤维细胞生长因子2(FGF2)、肿瘤坏死因子-α(TNF-α)水平;采用电化学发光法检测血清中白细胞介素-6(IL-6)水平;采用Pearson相关性分析检验糖尿病足病人血清DLX6-AS1、miR-346的表达水平与炎性因子的相关性;采用logistic回归分析研究影响糖尿病足发生的相关因素。结果糖尿病组、糖尿病足组病人VCAM-1、FGF2、TNF-α、IL-6水平高于对照组(P<0.05),糖尿病组与糖尿病足组比较差异有统计学意义(P<0.05);对照组DLX6-AS1的表达量是1.01±0.13,低于糖尿病组、糖尿病足组DLX6-AS1的表达量2.16±0.25、3.20±1.03(P<0.05);对照组miR-346的表达水平是1.00±0.16,高于糖尿病组、糖尿病足组miR-346的表达水平0.73±0.11、0.41±0.08(P<0.05),糖尿病组与糖尿病足组相比差异有统计学意义(P<0.05);随着糖尿病足病人Wagner分级增加DLX6-AS1的表达水平显著升高(P<0.05),miR-346的表达水平显著降低(P<0.05);DLX6-AS1的表达与FGF2、TNF-α、IL-6呈正相关(P<0.05),而miR-346与之呈负相关(P<0.05);DLX6-AS1与miR-346表达量是影响糖尿病足发生的危险因素(P<0.05)。结论糖尿病足病人血清中DLX6-AS1表达量升高,而miR-346表达量降低,二者可能参与糖尿病足发生及发展过程,并可能作为临床诊断的分子标志物。     

Alterations of epithelial permeability by Helicobacter and IL-1beta in vitro: protective effect of rebamipide

Infection with Helicobacter increases the transcellular passage of macromolecules across the epithelium, and this effect can be prevented by a gastroprotective agent rebamipide. The aim was to gain insight into the mechanisms involved. The HT29-19A intestinal epithelial cells grown on microporous filters as monolayers were incubated in the presence or absence of rebamipide (1 or 2 mM) with: (1) suspension of a wild H. pylori strain, (2) IL-1 (0.5 ng/ml) + IFN- (2 units/ml). After incubation, the monolayers were submitted to evaluation of apoptosis by using the apoptotic cell death detection ELISA kit and to assessment of epithelial permeability in Ussing chamber where the ionic conductance (G), fluxes of mannitol (J_Man) and of horseradish peroxidase in both intact (J_HRPi)- and degraded (J_D) form, were measured. H. pylori increased the intact HRP fluxes across the barrier (J_HRPi = 17 ± 20 vs 97 ± 70 ng/hr/cm², P < 0.007), an effect prevented by rebamipide (J_HRPi = 33 ± 34 ng/hr/cm², P < 0.006). IL-1 increased the ionic conductance (G = 5.5 ± 1.0 and 21.0 ± 7.0 mS/cm², P < 0.006), the intact HRP fluxes (J_HRPi = 18 ± 15 and 476 ± 344 ng/hr/cm², P < 0.006), and the apoptotic index of the cells (AI = 1 ± 0 vs 3.7 ± 0.8), all effects prevented by rebamipide (G = 12 ± 4.9 mS/cm², J_HRPi = 79 ± 38, AI = 1.6 ± 0.6, P < 0.03 as compared to IL--treated cells). In basal conditions, rebamipide increased the integrity of the barrier (G = 7.5 ± 2.3 vs 6.0 ± 1.8 mS/cm² for controls, P < 0.007). In conclusion, H. pylori as well as IL-1, may alter epithelial permeability and rebamipide may exert its protective effect on gastric mucosa by reinforcing the epithelial barrier in normal conditions and by counteracting the deleterious effect of Helicobacter pylori and IL-1 on macromolecular permeability.     

毛细管电泳测定micro RNA346基因多态性的方法研究

目的建立micro RNA346基因多态性的毛细管电泳（CE）测定方法。方法采用血液/细胞/组织基因组DNA提取试剂盒提取血清样品基因组DNA，PCR扩增micro RNA346目的基因，BciT130Ⅰ限制性内切酶酶切，产物用CE测定。对CE筛分介质质量浓度、分离电压等参数进行了优化。在优化的条件下（筛分介质质量浓度为10 g/L分离电压为12 kV)，分别测定类风湿性关节炎患者和正常人血清样品micro RNA346酶切产物，并对其进行基因分型。结果在优化的CE实验条件下（筛分介质质量浓度为10 g/L，分离电压为12 kV，25 min内可完成micro RNA346基因酶切产物检测。方法日内相对标准偏差（RSD）为0.43%～0.63%，日间RSD为1.49%～1.56%。用该法测定了96份类风湿性关节炎患者样品和43份正常人样品，结果均为micro RNA346 Ⅰ型，未发现micro RNA346 Ⅱ型。结论本研究建立的方法操作简单，具有高效、快速、样品用量少、自动化程度高等优点，适用于micro RNA类小分子RNA基因多态性的测定。     

Characterization of a Lineage C.36 SARS-CoV-2 Isolate with Reduced Susceptibility to Neutralization Circulating in Lombardy,Italy

SARS-CoV-2 spike is evolving to maximize transmissibility and evade the humoral response. The massive genomic sequencing of SARS-CoV-2 isolates has led to the identification of single-point mutations and deletions, often having the recurrence of hotspots, associated with advantageous phenotypes. We report the isolation and molecular characterization of a SARS-CoV-2 strain, belonging to a lineage (C.36) not previously associated with concerning traits, which shows decreased susceptibility to vaccine sera neutralization.