In vitro screening of Fe2+‐chelating effect by a Fenton's reaction–luminol chemiluminescence system

In vitro screening of a Fe²⁺‐chelating effect using a Fenton's reaction–luminol chemiluminescence (CL) system is described. The luminescence between the reactive oxygen species generated by the Fenton's reaction and luminol was decreased on capturing Fe²⁺ using a chelator. The proposed method can prevent the consumption of expensive seed compounds (drug discovery candidates) owing to the high sensitivity of CL detection. Therefore, the assay could be performed using small volumes of sample solution (150 μL) at micromolar concentrations. After optimization of the screening conditions, the efficacies of conventional chelators such as ethylenediaminetetraacetic acid (EDTA), diethylentriaminepentaacetic acid (DETAPAC), deferoxamine, deferiprone and 1,10‐phenanthroline were examined. EC₅₀ values for these compounds (except 1,10‐phenanthroline) were in the range 3.20 ± 0.87 to 9.57 ± 0.64 μM (n = 3). Rapid measurement of the Fe²⁺‐chelating effect with an assay run time of a few minutes could be achieved using the proposed method. In addition, the specificity of the method was discussed. Copyright © 2014 John Wiley & Sons, Ltd.     

HgCl2对泥鳅精子运动的影响

为了检测不同浓度的HgCl2对泥鳅Misgurnus anguillicaudatus精子运动的影响,以泥鳅精子为实验材料,用含终浓度分别为0(对照)、1、5、10、15和20 μmoL/L的HgCl2待测液分别孵育0、2、4和6 h后激活,激活后立即在显微镜(Olympus IX81)下观察精子运动参数.为进一步探讨HgCl2对泥鳅精子运动影响的机制,用终浓度为20μmol/L的HgCl2保存液孵育泥鳅精子10 min,以含终浓度分别为0、0.1、1、10 mmol/L的2-巯基乙醇和20μmol/LHgCl2混合液为激活液激活.激活后立即在显微镜下观察精子运动,发现2-巯基乙醇町逆转HgCl2对泥鳅精子的抑制作用,为探讨HgCl2对泥鳅精子运动影响的可能机制提供参考.     

脐带华通胶间充质干细胞向Flkl阳性细胞分化的研究

目的：诱导脐带华通胶间充质干细胞向Flk1阳性细胞分化。方法：胶原酶法分离培养脐带华通胶间充质干细胞,第3代细胞以含2-巯基乙醇的分化培养基培养,应用RT-PCR和流式细胞仪从mRNA和蛋白水平检测Flk1阳性细胞分化水平。结果：脐带华通胶间充质干细胞Flk1mRNA及蛋白表达极低,分化培养基培养后表达上调,48h达高峰（P〈0.05）,之后表达降低。结论：2-巯基乙醇可诱导脐带华通胶间充质干细胞向Flk1阳性细胞分化,为从中分选Flk1阳性细胞进行进一步研究提供了依据     

Function and dysfunction of leucine-rich repeat kinase 2 (LRRK2): Parkinson’s disease and beyond

Mutations in leucine-rich repeat kinase 2 (LRRK2) are the most common cause of familial Parkinson’s disease (PD). As such, functions and dysfunctions of LRRK2 in PD have been the subject of extensive investigation. In addition to PD, increasing evidence is suggesting that LRRK2 is associated with a wide range of diseases. Genome-wide association studies have implicated LRRK2 in Crohn’s disease (CD) and leprosy, and the carriers with pathogenic mutations of LRRK2 show increased risk to develop particular types of cancer. LRRK2 mutations are rarely found in Alzheimer’s disease (AD), but LRRK2 might play a part in tauopathies. The association of LRRK2 with the pathogenesis of apparently unrelated diseases remains enigmatic, but it might be related to the yet unknown diverse functions of LRRK2. Here, we reviewed current knowledge on the link between LRRK2 and several diseases, including PD, AD, CD, leprosy, and cancer, and discussed the possibility of targeting LRRK2 in such diseases. [BMB Reports 2015; 48(5): 243-248]     

脐带华通胶间充质干细胞向Flk1 阳性细胞分化的研究*

目的:诱导脐带华通胶间充质干细胞向Flk1阳性细胞分化。方法:胶原酶法分离培养脐带华通胶间充质干细胞,第3代细胞以含2-巯基乙醇的分化培养基培养,应用RT-PCR和流式细胞仪从mRNA和蛋白水平检测Flk1阳性细胞分化水平。结果:脐带华通胶间充质干细胞Flk1mRNA及蛋白表达极低,分化培养基培养后表达上调,48h达高峰(P<0.05),之后表达降低。结论:2-巯基乙醇可诱导脐带华通胶间充质干细胞向Flk1阳性细胞分化,为从中分选Flk1阳性细胞进行进一步研究提供了依据。     

Type 2 diabetes mellitus: An unusual association with Down's syndrome

Down''s syndrome (DS) is known to be associated with autoimmune disease including type 1 diabetes. To the best of our knowledge, there are no reports of DS with type 2 diabetes mellitus in the literature. We hereby report two cases of DS with type 2 diabetes.     

敲除NDRG2 基因的AD模型小鼠的构建与鉴定

目的:构建与鉴定NDRG2基因全身敲除的阿尔茨海默症(AD)小鼠模型。方法:将NDRG2-/-、APP/PS1进行饲养杂交繁殖,将通过PCR技术鉴定基因型为NDRG2+/-APP/PS1的子一代小鼠再与NDRG2-/-小鼠回交获得子二代小鼠,提取子二代小鼠的基因组DNA再利用PCR方法:扩增NDRG2和APP/PS1基因片段并进行琼脂糖凝胶电泳检测获得4种基因型的小鼠。结果:选取基因型为NDRG2-/-APP/PS1的小鼠即为全身NDRG2敲除且淀粉样蛋白基因过表达的阿尔茨海默症模型小鼠。应用PCR方法:鉴定NDRG2基因全身敲除的AD小鼠模型。成功获得NDRG2基因全身敲除的AD小鼠,该基因型小鼠有繁殖能力,其繁殖符合孟德尔遗传规律。结论:成功构建NDRG2基因敲除的阿尔茨海默症模型小鼠,为进一步研究NDRG2基因在阿尔茨海默症病理发展过程中的作用机制及新的治疗方法的研究提供模型基础。     

The Parkinson's disease protein LRRK2 impairs proteasome substrate clearance without affecting proteasome catalytic activity

Leucine-rich repeat kinase 2 (LRRK2) mutations are the most common known cause of Parkinson''s disease (PD). The clinical features of LRRK2 PD are indistinguishable from idiopathic PD, with accumulation of α-synuclein and/or tau and/or ubiquitin in intraneuronal aggregates. This suggests that LRRK2 is a key to understanding the aetiology of the disorder. Although loss-of-function does not appear to be the mechanism causing PD in LRRK2 patients, it is not clear how this protein mediates toxicity. In this study, we report that LRRK2 overexpression in cells and in vivo impairs the activity of the ubiquitin-proteasome pathway, and that this accounts for the accumulation of diverse substrates with LRRK2 overexpression. We show that this is not mediated by large LRRK2 aggregates or sequestration of ubiquitin to the aggregates. Importantly, such abnormalities are not seen with overexpression of the related protein LRRK1. Our data suggest that LRRK2 inhibits the clearance of proteasome substrates upstream of proteasome catalytic activity, favouring the accumulation of proteins and aggregate formation. Thus, we provide a molecular link between LRRK2, the most common known cause of PD, and its previously described phenotype of protein accumulation.     

女贞子提取物对帕金森病大鼠神经炎性反应及内质网应激PERK/ATF4通路的影响

摘要 目的：探讨与分析女贞子提取物对帕金森病大鼠神经炎性反应及内质网应激蛋白激酶R样内质网激酶（PERK）/转录活化因子4（ATF4）通路的影响。方法：采用单侧注射6-羟基多巴胺（6-OHDA）毁损法建立帕金森病大鼠模型,将造模成功的大鼠36只随机平分为三组-模型组、左旋多巴组与女贞子组,每组各12只。左旋多巴组与女贞子组分别灌胃0.5 mL的左旋多巴、女贞子提取物,模型组给予无菌蒸馏水0.5 mL,2次/d,连续给药4周,检测大鼠神经炎性反应、内质网应激PERK/ATF4通路相关蛋白表达变化情况。结果：左旋多巴组与女贞子组治疗第2周、第4周的探究性反应次数显著高于模型组（P<0.05）,女贞子组与左旋多巴组对比也有显著提高（P<0.05）。左旋多巴组与女贞子组治疗第2周、第4周的脑组织伊文思蓝含量显著低于模型组（P<0.05）,女贞子组与左旋多巴组对比也有显著降低（P<0.05）。左旋多巴组与女贞子组治疗第2周、第4周的血清丙二醛、白介素-1β、肿瘤坏死因子-α、一氧化氮含量显著低于模型组（P<0.05）,女贞子组与左旋多巴组对比也有显著降低（P<0.05）。左旋多巴组与女贞子组治疗第2周、第4周的黑质-纹状体组织PERK蛋白、ATF4蛋白相对表达水平显著低于模型组（P<0.05）,女贞子组与左旋多巴组对比也有显著降低（P<0.05）。结论：女贞子提取物在帕金森病大鼠的应用能改善神经功能,降低脑组织伊文思蓝含量,还可抑制PERK/ATF4通路的激活,降低血清丙二醛、白介素-1β、肿瘤坏死因子-α、一氧化氮含量,从而持续发挥脑保护作用。     

地塞米松对HepG2细胞分泌载脂蛋白的影响

以培养的人肝癌细胞系HepG2为研究对象,采用“冻干浓缩培养液载脂蛋白测定法”,考察了地塞米松对HepG2细胞载脂蛋白(apolipoprotein,apo)AⅠ、AⅡ、CⅢ、B100及E分泌的影响.结果表明：地塞米松对HepG2细胞apoAⅠ和apoE的分泌有促进作用,对apoAⅡ、apoB100和apoCⅢ的分泌有抑制作用,且这种作用随地塞米松浓度的增加而增强.当培养液中地塞米松的浓度为5.5×10^－5mol/L时,apoAⅠ和apoE的分泌分别增加36.6%和49.4%（P＜0.01）,apoAⅡ、apoB100和apoCⅢ的分泌分别减少38.9%、31.9%和29.8%（P＜0.01）.     

The inactivation of thymidylate synthase by periodate

Thymidylate synthase from methotrexate-resistant Lactobacillus casei rapidly lost about 90% of its catalytic activity when incubated with an equimolar concentration of IO4- at 0 degree C. Nearly complete inhibition resulted when the IO4- concentration was twice the enzyme concentration or higher. The inhibition reaction appeared to be pseudo-first-order with respect to enzyme when IO4- was in excess. The substrate dUMP, the product dTMP, and inorganic phosphate all protected the enzyme from inactivation by IO4-, with the order of effectiveness: dUMP greater than dTMP greater than phosphate. Deoxyuridine, which is not a substrate, did not protect the enzyme. Titrations with dithiobis(2-nitrobenzoate) (DTNB) showed that approximately 1.5 titratable SH groups were lost when thymidylate synthase was completely inhibited by IO4-. Essentially no reactivation occurred when periodate-inhibited enzyme was dialyzed against buffered 2-mercaptoethanol (ME) or dithiothreitol (DTT). Enzyme that had been treated with p-hydroxymercuribenzoate, DTNB, or methylmethanethiosulfonate prior to treatment with periodate could be completely reactivated with ME or DTT.     

Morgan's Canon: Is it still a useful rule of thumb?

For over 100 years, Morgan's Canon has served as the criterion for distinguishing what appears to be complex cognitive processes shown by animals from simpler associative learning processes (Pavlovian and instrumental conditioning). Morgan's ( 1894 ) canon states “In no case is an animal activity to be interpreted in terms of higher psychological processes if it can be fairly interpreted in terms of processes which stand lower in the scale of psychological evolution and development.” In the present article, several examples are provided in which complex human‐like processes are proposed to have been demonstrated but the judicious use of Morgan's Canon suggests that simpler mechanisms may be sufficient to account for the behavior. The use of Morgan's Canon is not meant to reduce behavior to its lowest common denominator but rather to challenge investigators to develop procedures that can distinguish between simple behavioral principles and the more complex processes that cannot be explained in terms of genetics or simple conditioning. Whatever the results of these experiments, they should help identify the underlying processes and mechanisms involved.