Detection of Protein A Produced by Staphylococcus aureus with a Fiber-optic-based Biosensor

Staphylococcus aureus is a pathogen important in causing human infections and intoxication. A sensitive fiber-optic that produces evanescent waves was developed for the detection of protein A, a product secreted only by S. aureus. In the immunosensor, a 40-mV argon-ion laser that generated laser light at 488 nm was used together with plastic optical fiber and antibodies to protein A were physically adsorbed onto the fiber. The principle of the detection involved a sandwich immunoassay with fluorescein isothiocyanate conjugated with anti-(protein A) immunoglobulin G to produce signals of the antigen-antibody reaction. The detection limit was 1 ng of protein A per milliliter. The fiber-optic immunosensor could be used for rapid and specific detection of S. aureus in clinical specimens and foods.     

Exogenous neuritin treatment improves survivability and functions of Schwann cells with improved outgrowth of neurons in rat diabetic neuropathy

Pathogenesis and treatment for diabetic neuropathy are still complex. A deficit of neurotrophic factors affecting Schwann cells is a very important cause of diabetic neuropathy. Neuritin is a newly discovered potential neurotrophic factor. In this study, we explored the effect of exogenous neuritin on survivability and functions of diabetic Schwann cells of rats with experimental diabetic neuropathy. Diabetic neuropathy was induced in rats. 12‐week diabetic rats contrasted with non‐diabetic normal rats had decreased levels of serum neuritin and slowed nerve conduction velocities (NCVs). Schwann cells isolated from these diabetic rats and cultured in high glucose showed reduced cell neuritin mRNA and protein and supernatant neuritin protein, increased apoptosis rates, increased caspase‐3 activities and progressively reduced viability. In contrast, exogenous neuritin treatment reduced apoptosis and improved viability, with elevated Bcl‐2 levels (not Bax) and decreased caspase‐3 activities. Co‐cultured with diabetic Schwann cells pre‐treated with exogenous neuritin in high glucose media, and diabetic DRG neurons showed lessened decreased neurite outgrowth and supernatant NGF concentration occurring in co‐culture of diabetic cells. Exogenous neuritin treatment ameliorated survivability and functions of diabetic Schwann cells of rats with diabetic neuropathy. Our study may provide a new mechanism and potential treatment for diabetic neuropathy.     

社区2型糖尿病患者血糖自我管理水平调查及并发糖尿病周围神经病变的影响因素分析

摘要 目的：调查社区2型糖尿病（T2DM）患者血糖自我管理水平及分析T2DM并发糖尿病周围神经病变（DPN）的影响因素。方法：于2016年6月~2017年6月期间采用整群随机抽样法随机抽取苏州市6个社区符合纳排标准的539例T2DM患者进行问卷调查,了解社区T2DM患者血糖自我管理水平情况,对患者进行体格检查并检测血生化指标,统计社区T2DM患者的DPN发生情况,采用多因素logistic回归分析T2DM患者并发DPN的影响因素。结果：本次研究共发放调查问卷539份,实际回收531份,其中T2DM并发DPN者86例,根据是否并发DPN将所有入选患者分为DPN组（n=86）和无DPN组（n=445）。社区T2DM患者的糖尿病自我管理行为量表（SDSCA）平均得分率为（49.38±5.23）%,DPN组和无DPN组在病程、体质量指数（BMI）、腰围、糖化血红蛋白（HbAlc）、空腹胰岛素（FINS）、低 密 度 脂 蛋 白（LDL-C）、血 尿 素 氮（BUN）、血肌酐（Cr）、合并外周动脉疾病（PAD）、合并糖尿病视网膜病变（DR）中比较差异有统计学意义（P<0.05）。多因素logistic回归分析结果显示：病程≥7年、HbAlc≥8 mmol/L、合并PAD、合并DR、BMI≥25 kg/m²是社区 T2DM患者并发DPN的危险因素（P<0.05）。结论：苏州市6个社区的T2DM患者血糖自我管理水平较低,且T2DM并发DPN的概率较高,病程≥7年、HbAlc≥8 mmol/L、合并PAD、合并DR、BMI≥25 kg/m²均是社区 T2DM患者并发DPN的危险因素,临床可对上述危险因素采取积极有效的措施,以有效降低T2DM并发DPN的发生率。     

The investigation of therapeutic potential of oxytocin and liraglutide on vincristine‐induced neuropathy in rats

The aim of this study was to assess the therapeutic potential of oxytocin and liraglutide (LIR), a GLP‐1 analogue, in a rat model of vincristine‐induced neuropathy. Rats were injected with vincristine (VCR) at a dose of 4 mg/kg twice a week for 5 weeks. The VCR‐administered rats were divided into three groups and received saline, oxytocin, or liraglutide simultaneously with VCR. After the treatment period, electrophysiological, biochemical, histological, and immunohistochemical investigations were performed. Electromyography (EMG) recordings demonstrated significant alterations in the VCR + saline group (p < .001). Also, motor performance was decreased in the VCR + saline group (p < .05). Histologically, the axonal diameter was decreased in all groups. VCR + saline group showed significantly increased lipid peroxidation and decreased nerve growth factor (NGF) expression. However, the administration of oxytocin and liraglutide significantly prevented the EMG alterations, lipid peroxidation, and reduction in neuronal NGF expression. On the basis of these findings, oxytocin and liraglutide may be considered as potential agents for the prevention of VCR‐induced neuropathy.     

Identification of novel inhibitors for the tyrosyl-DNA-phosphodiesterase 1 (Tdp1) mutant SCAN1 using virtual screening

Spinocerebellar ataxia syndrome with axonal neuropathy (SCAN1) is a debilitating neurological disease that is caused by the mutation the Tyrosyl-DNA phosphodiesterase 1 (TDP1) DNA repair enzyme. The crucial His493 in TDP1′s binding site is replaced with an arginine amino acid residue rendering the enzyme dysfunctional. A virtual screen was performed against the homology model of SCAN1 and seventeen compounds were identified and tested in a novel SCAN1 specific biochemical assay. Six compounds showed activity with IC₅₀ values between 3.5 and 25.1 µM. The most active ligand 5 (3.5 µM) is a dicoumarin followed by a close structural analogue 6 at 6.0 µM. A less potent series of β-carbolines (14 and 15) was found with potency in the mid-teens. According to molecular modelling an excellent fit for the active ligands into the binding pocket is predicted. To the best of our knowledge, data on inhibitors of the mutant form of TDP1 has not been reported previously. The virtual hits were also tested for wild type TDP1 activity and all six SCAN1 inhibitors are potent for the former, e.g., ligand 5 has a measured IC₅₀ at 99 nM.In the last decade, TDP1 is considered as a promising target for adjuvant therapy against cancer in combination with Topoisomerase 1 poisons. The active ligands are mostly non-toxic to cancer cell lines A-549, T98G and MCF-7 as well as the immortalized WI-38 human fetal lung cells. Furthermore, ligands 5 and 7, show promising synergy in conjunction with topotecan, a clinically used topoisomerase 1 anticancer drug. The active ligands 5, 7, 14 and 15 have a good balance of the physicochemical properties required for oral bioavailability making the excellent candidates for further development.     

神经节苷脂钠对缺血性脑卒中小鼠空间学习记忆能力的影响

摘要 目的：探讨神经节苷脂钠对缺血性脑卒中小鼠空间学习记忆能力的影响。方法：缺血性脑卒中小鼠模型(n=42)随机分为三组-模型组、氟西汀组与神经节苷脂钠组,每组14只小鼠。氟西汀组、神经节苷脂钠组、对照组分别给予10 mg/kg氟西汀与10 mg/kg神经节苷脂钠、等剂量生理盐水腹腔注射,1次/d,持续28 d。结果：氟西汀组、神经节苷脂钠组给药第7 d、14 d、28 d的逃避潜伏期、改良神经损伤严重程度评分(Modified neurological severity score,mNSS)低于模型组(P<0.05),穿越平台次数高于模型组(P<0.05),神经节苷脂钠组与氟西汀组对比差异有统计学意义(P<0.05)。氟西汀组、神经节苷脂钠组给药第28 d的海马组织B淋巴细胞瘤-2基因(B-cell lymphoma-2,Bcl-2)蛋白相对表达水平高于模型组(P<0.05),BCL2-Associated X(Bax)蛋白相对表达水平、脑卒中相对面积低于模型组(P<0.05),神经节苷脂钠组与氟西汀组对比差异有统计学意义(P<0.05)。结论：神经节苷脂钠在缺血性脑卒中小鼠的应用能促进恢复空间学习记忆能力,缓解神经损伤,抑制海马组织神经元细胞的凋亡,降低脑卒中面积。     

丁苯酞对缺血性脑卒中大鼠记忆能力及海马5-HT1A受体和PKA信号通路活化的影响

目的:研究丁苯酞对缺血性脑卒中大鼠学习和记忆能力的影响和大鼠海马5-HT1A受体和PKA信号通路的调控作用。方法:将雄性SD大鼠随机分为假手术组、模型组和丁苯酞组(n=15)。丁苯酞组大鼠建立大脑中动脉闭塞模型,并按照每天60 mg/kg的剂量灌胃丁苯酞,假手术组和模型组灌胃等体积的玉米油,共给药2周。治疗完成后对各组大鼠进行神经功能缺损评估和Morris水迷宫测试(n=15)。通过磁共振成像(MRI)检测梗塞区域(n=15)。ELISA法检测海马组织PKA激酶活性(n=6)。使用钙检测试剂盒测定海马组织的细胞内[Ca²⁺]浓度(n=6)。Western blot检测海马组织中5-羟色胺(1A)受体(5-HT1A)、谷氨酸N-甲基-D-天冬氨酸受体1(NMDA1)和α-氨基-3-羟基-5-甲基-4-异恶唑丙酸受体1(AMPA1)的表达(n=6)。结果:与模型组相比:丁苯酞组大鼠的逃避潜伏期显著降低,而穿越平台次数显著升高(P<0.05);大鼠的神经功能缺损评分和脑梗死体积较显著降低(P<0.05);大鼠的PKA激酶活性和细胞内[Ca²⁺]浓度显著升高(P<0.05);丁苯酞组大鼠的5-HT1A蛋白相对表达量显著降低,而AMPA1和NMDA1的磷酸化水平显著升高(P<0.05)。结论:丁苯酞可改善缺血性脑卒中大鼠的学习和记忆能力,下调海马5-HT1A受体活性并激活PKA信号通路。     

Genistein-3′-sodium sulfonate Attenuates Neuroinflammation in Stroke Rats by Down-Regulating Microglial M1 Polarization through α7nAChR-NF-κB Signaling Pathway

Microglial M1 depolarization mediated prolonged inflammation contributing to brain injury in ischemic stroke. Our previous study revealed that Genistein-3′-sodium sulfonate (GSS) exerted neuroprotective effects in ischemic stroke. This study aimed to explore whether GSS protected against brain injury in ischemic stroke by regulating microglial M1 depolarization and its underlying mechanisms. We established transient middle cerebral artery occlusion and reperfusion (tMCAO) model in rats and used lipopolysaccharide (LPS)-stimulated BV2 microglial cells as in vitro model. Our results showed that GSS treatment significantly reduced the brain infarcted volume and improved the neurological function in tMCAO rats. Meanwhile, GSS treatment also dramatically reduced microglia M1 depolarization and IL-1β level, reversed α7nAChR expression, and inhibited the activation of NF-κB signaling in the ischemic penumbra brain regions. These effects of GSS were further verified in LPS-induced M1 depolarization of BV2 cells. Furthermore, pretreatment of α7nAChR inhibitor (α-BTX) significantly restrained the neuroprotective effect of GSS treatment in tMCAO rats. α-BTX also blunted the regulating effects of GSS on neuroinflammation, M1 depolarization and NF-κB signaling activation. This study demonstrates that GSS protects against brain injury in ischemic stroke by reducing microglia M1 depolarization to suppress neuroinflammation in peri-infarcted brain regions through upregulating α7nAChR and thereby inhibition of NF-κB signaling. Our findings uncover a potential molecular mechanism for GSS treatment in ischemic stroke.     

Phagocytosis of degenerating myelin in transected feline optic nerve: an immunohistochemical study

The phagocytic activity of neuroglial cells in adult feline degenerating optic nerve was investigated by immunocytochemistry at both light and electron microscopy levels. Degeneration was initiated by unilateral eye enucleation and the segment distal to the transection showing true Wallerian degeneration was examined. Following enucleation, twelve adult domestic cats were examined over a period of seven to 215 days. All cases showed slow clearance of myelin debris and absence of proliferating monocytes throughout the post-enucleation period. All phagocytic cells present were neuroglial cells, and many of these cells expressed oligodendroglial antigens. These findings demonstrate the persistence of an active population of oligodendrocytes that might play an additional functional role during Wallerian degeneration of feline optic nerve.     

Diurnal and circadian regulations by three melatonin receptors in the brain and retina of olive flounder Paralichthys olivaceus: profiles following exogenous melatonin

To establish the molecular basis of circadian rhythm control by melatonin receptors (MTs), we investigated the mitochondrial ribonucleic acid (mRNA) expressions of three types of MTs in different tissues of the olive flounder (Paralichthys olivaceus). All three types of MT mRNAs were expressed in the neural tissues, while MT1 mRNA was expressed in the peripheral tissues and MT2 and MT3 mRNAs were weakly expressed or undetected in these tissues. We observed increased MT mRNA expression in the neural tissues at night under both light–dark (LD) and constant dark (DD) conditions. Although the melatonin-treated cultured pineal gland samples showed similar diurnal variations with high-MT mRNA expression levels at night compared to those of untreated cultured pineal gland samples, the expression levels were considerably higher in the melatonin-treated samples. The plasma melatonin level also significantly increased at night. Under DD conditions, the expression patterns of MT mRNAs were similar to those under the LD photocycle, but the peak was lower and the circadian change patterns were less clear. These findings reinforce the hypothesis that MTs are active in processing light information, and that these genes are regulated by the circadian clock and light, thus suggesting that MTs play an important role in daily and circadian variations in the brain and retina of olive flounders.