Overexpression of FNTB and the activation of Ras induce hypertrophy and promote apoptosis and autophagic cell death in cardiomyocytes

Farnesyltransferase (FTase) is an important enzyme that catalyses the modification of protein isoprene downstream of the mevalonate pathway. Previous studies have shown that the tissue of the heart in the suprarenal abdominal aortic coarctation (AAC) group showed overexpression of FTaseβ (FNTB) and the activation of the downstream protein Ras was enhanced. FTase inhibitor (FTI) can alleviate myocardial fibrosis and partly improve cardiac remodelling in spontaneously hypertensive rats. However, the exact role and mechanism of FTase in myocardial hypertrophy and remodelling are not fully understood. Here, we used recombinant adenovirus to transfect neonatal rat ventricular cardiomyocytes to study the effect of FNTB overexpression on myocardial remodelling and explore potential mechanisms. The results showed that overexpression of FNTB induces neonatal rat ventricular myocyte hypertrophy and reduces the survival rate of cardiomyocytes. FNTB overexpression induced a decrease in mitochondrial membrane potential and increased apoptosis in cardiomyocytes. FNTB overexpression also promotes autophagosome formation and the accumulation of autophagy substrate protein, LC3II. Transmission electron microscopy (TEM) and mCherry‐GFP tandem fluorescent‐tagged LC3 (tfLC3) showed that FNTB overexpression can activate autophagy flux by enhancing autophagosome conversion to autophagolysosome. Overactivated autophagy flux can be blocked by bafilomycin A1. In addition, salirasib (a Ras farnesylcysteine mimetic) can alleviate the hypertrophic phenotype of cardiomyocytes and inhibit the up‐regulation of apoptosis and autophagy flux induced by FNTB overexpression. These results suggest that FTase may have a potential role in future treatment strategies to limit the adverse consequences of cardiac hypertrophy, cardiac dysfunction and heart failure.     

Down‐regulation of Suv39h1 attenuates neointima formation after carotid artery injury in diabetic rats

Patients with diabetes have an increased risk of vascular complications. Suv39h1, a histone methyltransferase, plays a protective role against myocardial injury in diabetes. Herein, we intend to explore whether Suv39h1 could affect neointimal formation after vascular injury in diabetic rats and reveal the underlying mechanism. In this study, we generated adenovirus expressing Suv39h1 as well as lentivirus expressing Suv39h1‐targeting shRNA and evaluated the significance of Suv39h1 in vascular smooth muscle cells (VSMCs) under diabetic conditions. In vitro, we examined proliferative and migratory behaviours as well as the underlying signalling mechanisms in VSMCs in response to high glucose treatment. In vivo, we induced diabetes in SD rats with streptozocin and established the common carotid artery balloon injury model. Suv39h1 was found to be both necessary and sufficient to promote VSMC proliferation and migration under high glucose conditions. We observed corresponding changes in intracellular signalling molecules including complement C3 and phosphor‐ERK1/2. However, either up‐regulating or down‐regulating Suv39h1, phosphor‐p38 level was not significantly affected. Consistently, Suv39h1 overexpression led to accelerated neointima formation, while knocking down Suv39h1 reduced it following carotid artery injury in diabetic rats. Using microarray analyses, we showed that altering the Suv39h1 level in vivo dramatically altered the expression of myriad genes mediating different biological processes and molecular function. This study reveals the novel role of Suv39h1 in VSMCs of diabetes and suggests its potential role as a therapeutic target in diabetic vascular injury.     

Novel mutations in ADAMTS13 CUB domains cause abnormal pre-mRNA splicing and defective secretion of ADAMTS13

Hereditary thrombotic thrombocytopenic purpura (TTP) is an autosomal recessive thrombosis disorder, caused by loss-of-function mutations in ADAMTS13. Mutations in the CUB domains of ADAMTS13 are rare, and the exact mechanisms through which these mutations result in the development of TTP have not yet been fully elucidated. In this study, we identified two novel mutations in the CUB domains in a TTP family with an acceptor splice-site mutation (c.3569−1, G>A, intron 25) and a point missense mutation (c.3923, G>A, exon 28), resulting in a glycine to aspartic acid substitution (p.G1308D). In vitro splicing analysis revealed that the intronic mutation resulted in abnormal pre-mRNA splicing, and an in vitro expression assay revealed that the missense mutation significantly impaired ADAMTS13 secretion. Although both the patient and her brother displayed significantly reduced ADAMTS13 activity and increased levels of ultra-large VWF (ULVWF) multimers in plasma, only the female developed acute episodes of TTP. Our findings indicate the importance of the CUB domains for the protein stability and extracellular secretion of ADAMTS13.     

Formononetin,J1 and J2 have different effects on endothelial cells via EWSAT1‐TRAF6 and its downstream pathway

Formononetin is a natural isoflavone compound found mainly in Chinese herbal medicines such as astragalus and red clover. It is considered to be a typical phytooestrogen. In our previous experiments, it was found that formononetin has a two‐way regulatory effect on endothelial cells (ECs): low concentrations promote the proliferation of ECs and high concentrations have an inhibitory effect. To find a specific mechanism of action and provide a better clinical effect, we performed a structural transformation of formononetin and selected better medicinal properties for formononetin modifier J1 and J2 from a variety of modified constructs. The MTT assay measured the effects of drugs on human umbilical vein endothelial cell (HUVEC) activity. Scratch and transwell experiments validated the effects of the drugs on HUVEC migration and invasion. An in vivo assessment effect of the drugs on ovariectomized rats. Long‐chain non‐coding RNA for EWSAT1, which is abnormally highly expressed in HUVEC, was screened by gene chip, and the effect of the drug on its expression was detected by PCR after the drug was applied. The downstream factors and their pathways were analysed, and the changes in the protein levels after drug treatment were evaluated by Western blot. In conclusion, the mechanism of action of formononetin, J1 and J2 on ECs may be through EWSAT1‐TRAF6 and its downstream pathways.     

Structure Elucidation of Two New Norlignans from Anemone vitifolia and Their Anti‐Inflammatory Activities

Two new norlignans together with two known phenylpropanoids were isolated from the whole herb of Anemone vitifolia. All compounds were reported from this plant for the first time. The structures of these compounds were identified by comprehensive HR‐ESI‐MS, 1D and 2D NMR spectroscopic data analysis and comparison with literature data. Additionally, bioactivity study results showed that two new compounds have potential anti‐inflammatory activity. The plausible biosynthetic pathway for these compounds were also speculated in this article.     

Covalent–Organic Frameworks: Advanced Organic Electrode Materials for Rechargeable Batteries

Covalent–organic frameworks (COFs), featuring structural diversity, framework tunability and functional versatility, have emerged as promising organic electrode materials for rechargeable batteries and garnered tremendous attention in recent years. The adjustable pore configuration, coupled with the functionalization of frameworks through pre‐ and post‐synthesis strategies, enables a precise customization of COFs, which provides a novel perspective to deepen the understanding of the fundamental problems of organic electrode materials. In this review, a summary of the recent research into COFs electrode materials for rechargeable batteries including lithium‐ion batteries, sodium‐ion batteries, potassium‐ion batteries, and aqueous zinc batteries is provided. In addition, this review will also cover the working principles, advantages and challenges, strategies to improve electrochemical performance, and applications of COFs in rechargeable batteries.     

Simultaneous nitrogen and carbon removal in a packed A/O reactor: effect of C/N ratio on microbial community structure

In this research, a novel packed anoxic/oxic moving bed biofilm reactor (MBBR) was established to achieve high-organic matter removal rates, despite the carbon/nitrogen (C/N) ratio of 2.7–5.1 in the influent. Simultaneous nitrification–denitrification (SND) was investigated under a long sludge retention time of 104 days. The system exhibited excellent performance in pollutant removal, with chemical oxygen demand and total nitrogen (TN) enhanced to 93.6–97.4% and 34.4–60%, respectively. Under low C/N conditions, the nitrogen removal process of A/O MBBR system was mainly achieved by anaerobic denitrification. The increase of C/N ratio enhanced SND rate of the aerobic section, where dissolved oxygen was maintained at the range of 4–6 mg/L, and resulted in higher TN removal efficiency. The microbial composition and structures were analyzed utilizing the MiSeq Illumina sequencing technique. High-throughput pyrosequencing results indicated that the dominant microorganisms were Proteobacteria and Bacteroidetes at the phylum level, which contributes to the removal of organics matters. In the aerobic section, abundances of Nitrospirae (1.12–29.33%), Burkholderiales (2.15–21.38%), and Sphingobacteriales (2.92–11.67%) rose with increasing C/N ratio in the influent, this proved that SND did occur in the aerobic zone. As the C/N ratio of influent increased, the SND phenomenon in the aerobic zone of the system is the main mechanism for greatly improving the removal rate of TN in the aerobic section. The C/N ratio in the aerobic zone is not required to be high to exhibit good TN removal performance. When C/NH₄⁺ and C/TN in the aerobic zone were higher than 2.29 and 1.77, respectively, TN removal efficiency was higher than 60%, which means that carbon sources added to the reactor could be saved. This study would be vital for a better understanding of microbial structures within a packed A/O MBBR and the development of cost-efficient strategies for the treatment of low C/N wastewater.

     

去N端信号肽的水痘-带状疱疹病毒糖蛋白E的原核可溶性表达及其免疫原性评价

水痘-带状疱疹病毒(varicella zoster virus,VZV)糖蛋白E(glycoprotein E,gE)是VZV亚单位疫苗的主要候选蛋白,但目前原核表达系统制备的gE蛋白以包涵体形式为主,可溶性差。本研究采用去除第1～30氨基酸序列的VZV gE胞外域基因,将其与原核表达载体pET32a连接,并转化至感受态细胞BL21(DE3)中。使用异丙基-β-D-硫代半乳糖苷(Isopropylβ-D-thiogalactoside,IPTG)诱导表达,His-tag柱纯化重组gE蛋白,蛋白质印迹法(Western blot,WB)检测其特异性。用该重组gE蛋白免疫BALB/c小鼠制备多克隆抗体,酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)和间接免疫荧光法检测多克隆抗体效价及特异性。结果显示,BL21/pET32a-VZV gE工程菌可以表达可溶性重组gE蛋白,纯化后纯度约为90%。WB鉴定该重组蛋白具有良好的免疫反应性。ELISA检测显示小鼠抗VZV gE多克隆抗体效价＞1∶10 000,间接免疫荧光实验结果显示该抗体特异性较高。结果表明,本研究在原核表达系统中成功表达可溶性重组VZV gE蛋白,同时该蛋白具有较强的免疫原性,这为VZV gE亚单位疫苗的研制和大规模生产奠定了基础。     

房颤射频消融术后复发风险预测评分系统的建立及评价

目的:探讨心房颤动(房颤)患者射频消融术后复发的风险因素,并依此构建个性化的风险评分系统。方法:选取2017年1~8月行射频消融术的房颤患者154例作为研究对象,依据术后3个月的随访结果将患者分为复发组及未复发组,采用单因素分析和Logistic回归分析对各风险因素进行分析,构建其评分系统,采用Hosmer-Lemeshow拟合优度检验和ROC曲线下面积评价评分系统的准确度及区分度。结果:术后随访3个月的结果显示共37例(24.03%)房颤患者出现复发,房颤类型、病程、体质量指数(BMI)、左房前后径(LAD)、左房容积(LAV)及超敏C反应蛋白(hs-CRP)水平均是房颤复发的独立风险因素(P<0.05)。构建的风险评分系统得分为0~26分,Hosmer-Lemeshow拟合优度检验:x^2=7.520,P=0.482;ROC曲线下面积为0.864(95%CI:0.837~0.891),预测评分值为15分时,约登指数最大(0.605),此时的敏感度和特异度分别为77.3%和83.2%。结论:房颤患者射频消融术后的复发率较高,依据风险因素构建的风险评分系统具有较高的预测效率和区分能力,可作为房颤患者射频消融术后复发风险评估的参考工具。     

伤椎置钉联合短节段内固定与单纯短节段固定治疗胸腰椎爆裂性骨折的比较研究

目的:比较伤椎置钉联合短节段内固定与单纯短节段固定治疗胸腰椎爆裂性骨折的临床疗效、固定效果及其对患者炎症反应和脊髓损伤的影响。方法:选取2014年3月到2016年12月期间我院收治的胸腰椎爆裂性骨折患者94例,根据手术方法的不同将患者分为伤椎置钉组(40例)和短节段内固定组(44例)。短节段内固定组患者采用单纯后路短节段椎弓根螺钉内固定进行治疗,伤椎置钉组采用伤椎置钉联合后路短节段椎弓根螺钉内固定进行治疗。比较两组患者的手术时间、术中出血量、住院时间、伤椎前沿高度比、Cobb’s角、伤椎椎体楔形变角、视觉模拟评分(VAS)和Oswestry功能障碍指数(ODI),炎性因子指标、脊髓损伤指标及术后并发症。结果:伤椎置钉组的手术时间长于短节段内固定组(P<0.05),术后6个月、术后12个月伤椎置钉组的伤椎前沿高度比明显高于短节段内固定组,Cobb’s角、伤椎椎体楔形变角明显低于短节段内固定组(P<0.05),术前、术后1周、术后6个月、术后12个月两组患者的VAS评分和ODI比较差异无统计学意义(P>0.05),术后3 d两组患者血清中IL-1β、IL-6、IL-8、TNF-α和pNF-H、NSE、S100β、GFAP水平比较差异均无统计学意义(P>0.05)。随访期间两组患者均未出现严重并发症。结论:伤椎置钉联合后路短节段椎弓根螺钉内固定可有效改善胸腰椎爆裂性骨折患者的椎体高度、Cobb’s角和伤椎椎体楔形变角,并且不会增加脊髓损伤和机体的炎症反应。