微生物NAD合成酶及其抑制剂的研究进展

NAD(P)生物代谢在能量代谢,维持氧化还原稳态以及调节细胞寿命等许多细胞进程中有重要作用。因此,NAD生物合成途径的关键酶的抑制剂就成为备受关注的候选新药,如NAD合成酶抑制剂。本文对微生物中的NAD合成酶的催化活性特征,晶体结构,调控因子以及基于晶体结构的抑制剂设计方面进行了综述,以期为基于NAD的治疗领域打开新的思路。     

Transforming Activity of a Novel Mutant of HPV16 E6E7 Fusion Gene

An optimized recombinant HPV16 E6E7 fusion gene (HPV16 ofE6E7) was constructed according to codon usage for mammalian cell expression, and a mutant of HPV16 ofE6E7 fusion gene (HPV16 omfE6E7) was generated by site-directed mutagenesis at L57G, C113R for the E6 protein and C24G, E26G for the E7 protein for HPV16 ofE6E7 [patent pending (CN 101100672)]. The HPV16 omfE6E7 gene constructed in this work not only lost the transformation capability to NIH 3T3 cells and tumorigenicity in SCID mice, but also maintain...     

Interleukin-33/ST2 signaling promotes production of interleukin-6 and interleukin-8 in systemic inflammation in cigarette smoke-induced chronic obstructive pulmonary disease mice

Interleukin-33 is a newly described member of the interleukin-1 family. Recent research suggests that IL-33 is increased in lungs and plays a critical role in chronic airway inflammation in cigarette smoke-induced chronic obstructive pulmonary disease (COPD) mice. To determine the role of IL-33 in systemic inflammation, we induced COPD mice models by passive cigarette smoking and identified the IL-33 expression in bronchial endothelial cells and peripheral blood mononuclear cells (PBMCs) of them. After isolation, PBMCs were cultured and stimulated in vitro. We measured expressions of interleukin-6 and interleukin-8 in PBMCs in different groups. The expression of IL-33 in bronchial endothelial cells and PBMCs of COPD mice were highly expressed. Stimulated by cigarette smoke extract (CSE), the expression of IL-6 and IL-8 were induced and enhanced by IL-33. PBMCs of COPD mice produced more IL-6 and IL-8 stimulated by CSE and IL-33. Expression of IL-6 and IL-8 were decreased when stimulated by IL-33 together with soluble ST2. The mRNA production of ST2 in IL-33 stimulated PBMCs was increased. Being pretreated with several kinds of MAPK inhibitors, the secretions of IL-6 and IL-8 in PBMCs did not decrease except for the p38 MAPK inhibitor. We found that IL-33 could induce and enhance the expression of IL-6 and IL-8 in PBMCs of COPD mice via p38 MAPK pathway, and it is a promoter of the IL-6 and IL-8 production in systemic inflammation in COPD mice.     

Blockade effects of (Nphe1)Nociceptin(1-13)-NH(2) on anti-nociception induced by intrathecal administration of nociceptin in rats

The present study investigated the roles of the opioid-receptor-like (ORL1) receptor and its endogenous ligand nociceptin on nociception in the spinal cord of rats. Intrathecal administration of 10 nmol of nociceptin produced significant increases in hindpaw withdrawal latencies (HWLs) to thermal and mechanical stimulation. There were no significant changes of average maximum angles in inclined plane tests after intrathecal injection of 10 nmol of nociceptin in rats. The intrathecal nociceptin-induced increases in HWL were antagonized by intrathecal administration of (Nphe1)Nociceptin(1-13)-NH(2), a selective antagonist of ORL1 receptor, in a dose-dependent manner. The results demonstrated that ORL1 receptor is involved in the nociceptin-induced anti-nociceptive effect in the spinal cord of rats.     

O‐GlcNAcylation of protein kinase A catalytic subunits enhances its activity: a mechanism linked to learning and memory deficits in Alzheimer's disease

Alzheimer's disease (AD) is characterized clinically by memory loss and cognitive decline. Protein kinase A (PKA)‐CREB signaling plays a critical role in learning and memory. It is known that glucose uptake and O‐GlcNAcylation are reduced in AD brain. In this study, we found that PKA catalytic subunits (PKAcs) were posttranslationally modified by O‐linked N‐acetylglucosamine (O‐GlcNAc). O‐GlcNAcylation regulated the subcellular location of PKAcα and PKAcβ and enhanced their kinase activity. Upregulation of O‐GlcNAcylation in metabolically active rat brain slices by O‐(2‐acetamido‐2‐deoxy‐d ‐glucopyranosylidenamino) N‐phenylcarbamate (PUGNAc), an inhibitor of N‐acetylglucosaminidase, increased the phosphorylation of tau at the PKA site, Ser214, but not at the non‐PKA site, Thr205. In contrast, in rat and mouse brains, downregulation of O‐GlcNAcylation caused decreases in the phosphorylation of CREB at Ser133 and of tau at Ser214, but not at Thr205. Reduction in O‐GlcNAcylation through intracerebroventricular injection of 6‐diazo‐5‐oxo‐l ‐norleucine (DON), the inhibitor of glutamine fructose‐6‐phosphate amidotransferase, suppressed PKA‐CREB signaling and impaired learning and memory in mice. These results indicate that in addition to cAMP and phosphorylation, O‐GlcNAcylation is a novel mechanism that regulates PKA‐CREB signaling. Downregulation of O‐GlcNAcylation suppresses PKA‐CREB signaling and consequently causes learning and memory deficits in AD.     

磷脂酶A2亚型及其与男性生殖相关性研究

磷脂酶A2(phospholipase A2,PLA2)是参与细胞代谢活动、信号转导等过程的重要酶类之一,广泛分布于哺乳动物机体内。随着对磷脂酶A2研究的深入,目前已有30余种磷脂酶A2亚型被逐一鉴定,磷脂酶A2亚型与男性生殖相关性研究也有了新的发现。该文通过对磷脂酶A2的最新亚型分类及其特征、磷脂酶A2在男性生殖系统中的分布、磷脂酶A2参与精子顶体反应及其信号转导通路以及与男性生殖相关的磷脂酶A2亚型等内容进行分析,研究磷脂酶A2在男性生殖系统中发挥的作用及其机制,为男性不育的诊断和治疗提供理论依据和思路策略。     

绿色糖单孢菌-木素过氧化物酶的发酵工艺研究

本研究以一株中度耐热耐碱放线菌--绿色糖单孢菌(Saccharomonospora viridis)为研究对象,用16 L发酵罐对该菌进行了木素过氧化物酶(lignin peroxidases, LiP)的诱导发酵,确定了最适的产酶工艺条件:接种量为10%,C/N为1∶3,搅拌速度为250 r/min,通气量为5 L/min,通过控制通气量和调整搅拌转速,使溶氧维持在35%以上,此条件下绿色糖单孢菌较摇瓶实验提前将近24 h达到产酶高峰,酶活最高可达0.41 U/ml;同时在发酵罐中测定该菌株的生长曲线和代谢曲线以确定其发酵代谢规律.     

Quantitative determination of helicid in rat biosamples by liquid chromatography electrospray ionization mass spectrometry

A simple liquid chromatography electrospray ionization mass spectrometry (LC–ESI–MS) method with highly improved sensitivities for the determination of helicid in rat bile, urine, feces and most tissues was developed. The tissues and feces were firstly homogenized mechanically using deionized water as the media. Bile, urine, tissues and feces homogenates were extracted by liquid–liquid extraction with n-butyl alcohol for sample preparation. The subsequent analysis procedures were performed on a Shimadzu LCMS2010A system (electrospray ionization single quadrupole mass analyzer). A Luna C₁₈ column (150 mm × 2.00 mm, 5 μm) was used as the analytical column, while a mixture of acetonitrile and ammonium chloride water solution was used as the mobile phase. The proportions of mobile phase were changed timely according to gradient programs. Chlorinated adducts of molecular ions [M+Cl]^? at m/z 319.00 and 363.05 were used to quantify helicid and bergeninum (internal standard), respectively. The method was validated to be accurate, precise and rugged with good linearity. The proposed method was successfully applied to the preclinical tissue distribution and excretion studies of helicid in rats.     

Efficient Passivation of Hybrid Perovskite Solar Cells Using Organic Dyes with COOH Functional Group

Organic–inorganic halide perovskites are efficient absorbers for solar cells. Nevertheless, the trap states at the surfaces and grain boundaries are a detrimental factor compromising the device performance. Here, an organic dye (AQ310) is employed as passivator to reduce the trap states of the perovskites and promote better stability. The results demonstrate that the trap states of perovskite are minimized by the presence of AQ310's ?COOH group and the formation of coordination with under‐coordinated Pb²⁺ ions. The resulting carrier recombination time is prolonged and verified by the photoluminescence and open‐circuit voltage decay measurements. Consequently, the best average power conversion efficiency (PCE) of 19.43% is achieved for the perovskite solar cell (PSC) with AQ310 passivation, as compared with a low average PCE of 17.98% for the PSC without AQ310 passivation.