Expression and Purification of Membrane Proteins in Different Hosts

International Journal of Peptide Research and Therapeutics - Membrane proteins play important functions, such as cellular communication and transferring materials in the cell. Many membrane...     

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.

     

关节镜联合富血小板血浆对膝关节半月板损伤患者膝关节功能和生活质量的影响

目的:探讨关节镜联合富血小板血浆对膝关节半月板损伤患者膝关节功能和生活质量的影响。方法:选取2017年12月-2019年9月期间我院收治的膝关节半月板损伤患者80例,根据随机数字表法分为对照组(n=40)和研究组(n=40),对照组予以关节镜下修整手术治疗,研究组在对照组基础上联合富血小板血浆治疗,比较两组患者优良率、生活质量及视觉模拟评分量表(VAS)、Lysholm评分量表、美国西安大略和麦克马斯特大学关节炎指数(WOMAC)评分。记录两组治疗期间不良反应情况。结果:研究组治疗后3个月的膝关节功能优良率高于对照组(P0.05)。两组治疗前、治疗后1个月、治疗后3个月VAS、WOMAC评分均逐步降低,Lysholm评分逐步升高(P0.05);研究组治疗后1个月、治疗后3个月VAS、WOMAC评分低于对照组,Lysholm评分高于对照组(P0.05)。两组治疗后3个月SF-36各维度评分均较治疗前升高,且研究组高于对照组(P0.05)。两组不良反应发生率对比未见统计学差异(P0.05)。结论:关节镜联合富血小板血浆治疗膝关节半月板损伤患者,可促进膝关节功能的恢复,可有效缓解手术治疗后的疼痛症状,改善患者生活质量,具有较好的临床应用价值。     

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。     

穴位埋线联合益肾化痰方对多囊卵巢综合征患者T、E2、LH水平影响

摘要 目的：探讨穴位埋线联合益肾化痰方对多囊卵巢综合征患者睾酮(testosterone,T)、雌二醇(estradiol,E₂)、黄体生成素(luteinizing hormone,LH)水平影响。方法：选取2018年1月至2019年6月我院收治的120例多囊卵巢综合征患者作为研究对象,随机将其分为两组,对照组60例,给予益肾化痰方治疗,研究组60例,在对照组的基础上联合穴位埋线治疗。观察两组治疗后的疗效及不良反应率,检测两组治疗后子宫内膜厚度、成熟卵泡数以及治疗前后T、E₂、LH水平变化情况。结果：研究组治疗的总有效率为 98.33 %,显著高于对照组治疗的总有效率（85.0 %,P＜0.05）。两组治疗后子宫内膜厚度及成熟卵泡数比较,研究组明显高于对照组(P＜0.05)。两组治疗前T、E₂、LH水平指标比较差异无统计学意义（P>0.05）;两组治疗后T、E₂、LH水平指标比较差异有统计学意义（P<0.05）。对照组头痛3例,恶心4例,不良反应率为11.67 %;研究组头痛1例,不良反应率为1.67 %,两组患者不良反应率比较有统计学意义（P<0.05）。结论：埋线联合益肾化痰方治疗多囊卵巢综合征患者,能有效的提高治疗疗效,改善患者T、E2、LH水平,促进排卵,安全可靠,值得临床推广和应用。