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51.
Bingqing Xia Xurui Shen Yang He Xiaoyan Pan Feng-Liang Liu Yi Wang Feipu Yang Sui Fang Yan Wu Zilei Duan Xiaoli Zuo Zhuqing Xie Xiangrui Jiang Ling Xu Hao Chi Shuangqu Li Qian Meng Hu Zhou Yubo Zhou Xi Cheng Xiaoming Xin Lin Jin Hai-Lin Zhang Dan-Dan Yu Ming-Hua Li Xiao-Li Feng Jiekai Chen Hualiang Jiang Gengfu Xiao Yong-Tang Zheng Lei-Ke Zhang Jingshan Shen Jia Li Zhaobing Gao 《Cell research》2021,31(8):847-860
Cytokine storm and multi-organ failure are the main causes of SARS-CoV-2-related death. However, the origin of excessive damages caused by SARS-CoV-2 remains largely unknown. Here we show that the SARS-CoV-2 envelope (2-E) protein alone is able to cause acute respiratory distress syndrome (ARDS)-like damages in vitro and in vivo. 2-E proteins were found to form a type of pH-sensitive cation channels in bilayer lipid membranes. As observed in SARS-CoV-2-infected cells, heterologous expression of 2-E channels induced rapid cell death in various susceptible cell types and robust secretion of cytokines and chemokines in macrophages. Intravenous administration of purified 2-E protein into mice caused ARDS-like pathological damages in lung and spleen. A dominant negative mutation lowering 2-E channel activity attenuated cell death and SARS-CoV-2 production. Newly identified channel inhibitors exhibited potent anti-SARS-CoV-2 activity and excellent cell protective activity in vitro and these activities were positively correlated with inhibition of 2-E channel. Importantly, prophylactic and therapeutic administration of the channel inhibitor effectively reduced both the viral load and secretion of inflammation cytokines in lungs of SARS-CoV-2-infected transgenic mice expressing human angiotensin-converting enzyme 2 (hACE-2). Our study supports that 2-E is a promising drug target against SARS-CoV-2.Subject terms: Cell death, Molecular biology 相似文献
52.
Leucine dehydrogenase (LDH, EC 1.4.1.9) catalyzes the reversible deamination of branched-chain L-amino acids to their corresponding keto acids using NAD+ as a cofactor. LDH generally adopts an octameric structure with D4 symmetry, generating a molecular mass of approximately 400 kDa. Here, the crystal structure of the LDH from Pseudomonas aeruginosa (Pa-LDH) was determined at 2.5 Å resolution. Interestingly, the crystal structure shows that the enzyme exists as a dimer with C2 symmetry in a crystal lattice. The dimeric structure was also observed in solution using multiangle light scattering coupled with size-exclusion chromatography. The enzyme assay revealed that the specific activity was maximal at 60°C and pH 8.5. The kinetic parameters for three different amino acid and the cofactor (NAD+) were determined. The crystal structure represents that the subunit has more compact structure than homologs’ structure. In addition, the crystal structure along with sequence alignments indicates a set of non-conserved arginine residues which are important in stability. Subsequent mutation analysis for those residues revealed that the enzyme activity reduced to one third of the wild type. These results provide structural and biochemical insights for its future studies on its application for industrial purposes. 相似文献
53.
在人工气候室内比较研究了灰飞虱和褐飞虱在汕优63和秀水11上的种内和种间密度效应对其主要生物学参数的影响,结果表明两种稻飞虱均存在显著的种内竞争,主要表现为随着密度增加,若虫发育历期延长、若虫羽化率下降、雌成虫寿命缩短、每雌产卵量下降。种内密度效应与稻飞虱种类和寄主品种有显著互作关系,灰飞虱种内竞争较褐飞虱明显,二者在适宜寄主上种内竞争更为明显。同时,两种稻飞虱存在着显著的种间密度效应,主要表现为异种存在时的促进作用,即异种共存时若虫历期缩短、若虫羽化率提高、雌虫寿命延长、每雌产卵量增加。种间密度效应也与稻飞虱种类和寄主品种有显著互作关系,两种共存对灰飞虱的有利作用显著大于对褐飞虱的有利作用,在欠适宜寄主上种间互利效应更为明显。最后,本文对稻飞虱种内和种间密度效应的机制和进化意义进行了讨论。 相似文献
54.
Zhengquan Yang Chengliang Zhang Guojun Lian Shijie Dong Menghui Song Hengrong Shao Jingmei Wang Tao Zhong Zhenni Luo Shengnan Jin Chunming Ding 《Nucleic acids research》2022,50(13):7560
5′-Adenylated oligonucleotides (AppOligos) are widely used for single-stranded DNA/RNA ligation in next-generation sequencing (NGS) applications such as microRNA (miRNA) profiling. The ligation between an AppOligo adapter and target molecules (such as miRNA) no longer requires ATP, thereby minimizing potential self-ligations and simplifying library preparation procedures. AppOligos can be produced by chemical synthesis or enzymatic modification. However, adenylation via chemical synthesis is inefficient and expensive, while enzymatic modification requires pre-phosphorylated substrate and additional purification. Here we cloned and characterized the Pfu RNA ligase encoded by the PF0353 gene in the hyperthermophilic archaea Pyrococcus furiosus. We further engineered fusion enzymes containing both Pfu RNA ligase and T4 polynucleotide kinase. One fusion enzyme, 8H-AP, was thermostable and can directly catalyze 5′-OH-terminated DNA substrates to adenylated products. The newly discovered Pfu RNA ligase and the engineered fusion enzyme may be useful tools for applications using AppOligos. 相似文献
55.
荧光假单胞菌抗噬菌体菌株的选育 总被引:6,自引:2,他引:4
本实验从荧光假单胞菌(Pseudomonasfluorescens)AS—3菌株的不正常发酵液中分离到一种噬菌体,将其命名为PFAS。AS—3菌株能利用葡萄糖发酵产生D-异维生素C的前体物质2-酮基-D-葡萄糖酸。电镜观察表明PFAS噬菌体呈蝌蚪形,具有直径为66nm的六角形头部及长117nm的尾部。通过紫外线诱变及自然选育两种途径,配合简便有效的初筛方法,经多次分离、纯化、复筛最终在摇并发酵试验中获得6株产量稳定地高于对照敏感菌的抗噬菌体菌株,可望用于生产。 相似文献
56.
鼎湖山森林地表水水质状况分析 总被引:24,自引:4,他引:20
对鼎湖山生物圈保护区内两溪流水源水质状况进行了调查,结果发现:①鼎湖山地表水总体水质较好,符合地面水环境质量标准的I类水源水质标准。区内水体受污染程度低,绝大多数指标均符合饮用水卫生标准。②水体pH值较低,Al含量较高,总有机碳测定表现出整个水体受到一定程度的有机污染。水体中有害金属Mn和Pb含量略高,但都远低于饮用水卫生标准。③对地表水来源过程环节的水化学分析比较表明:大气降水、穿透水、土壤溶液(30cm层和80cm层)和地表水水样的pH值呈现“M”形变化。酸雨和土壤表层酸化是该区地表水pH值偏低的主要原因。地表水和30cm土壤溶液中的Al浓度分别是大气降水的5倍和8倍,地表水中的Al主要来源于酸雨对土壤的淋溶。地表水中的Na主要来源于大气降水。大气降水Pb浓度是地表水的17倍,林冠吸收富集和土壤固定吸附使地表水中的Pb大幅度降低。穿透水和土壤溶液中的Mn、K、Ca、Mg、Sr比大气降水和地表水浓度高,反映了元素被酸雨淋溶、活化和被植物、土壤吸收吸附的过程。从长远看,尽管在森林保存完好的地区,区域环境的恶化及酸雨对地表水水质的影响仍不容忽视。 相似文献
57.
58.
59.
流域季节性径流变化反映了年内水资源的动态特征。在以森林为主的流域中,森林变化和气候变异被普遍认为是影响流域水文过程的两大驱动因素。因此在全球气候变化背景下,研究流域森林恢复和气候变异对流域季节性径流的影响,可为协调区域碳-水关系和制订可持续的森林经营管理策略提供参考。选择鄱阳湖流域上游的平江流域为研究对象,根据流域历史森林覆盖率变化情况,将研究期划分为参考期(1961-1985)和森林恢复期(1986-2006),采用Mann-Kendall趋势分析研究流域长时期水文气象数据是否存在显著变化趋势。同时引入月干旱指数(潜在蒸散发和有效降雨的比率),将一年定义为能量限制季(1-6月)和水分限制季(7-12月),结合扩展的Budyko模型定量分析平江流域森林恢复和气候变异对季节性径流的相对贡献。在本研究流域整个研究期内(1961-2006),通过Mann-Kendall趋势分析发现,研究流域水分限制季径流呈现显著增加趋势,而能量限制季水文和气候变量变化趋势均不显著。其次,相较于参考期,流域森林恢复使能量限制季径流降低了11.71 mm/a (24.40%),使水分限制季径流增加了12.27 mm/a (17.23%)。同时,气候变异导致能量限制季径流减少了36.28 mm/a (75.60%),而使水分限制季径流增加了58.94 mm/a (82.77%)。上述研究结果表明,森林恢复对径流影响具有累积效应。森林恢复对季节性径流具有积极的调节作用,同时季节性径流对森林恢复的响应存在时间差,而且森林恢复对径流的影响在能量限制季和水分限制季具有相互抵消的作用,气候变异与森林恢复的影响效应类似。此外,本研究也证实,平江流域季节性径流变化主要是受气候变化主导,但森林恢复对季节性径流的贡献也不容忽视。 相似文献
60.
Dai Sik Ko Junho Kang Hye Jin Heo Eun Kyoung Kim Kihun Kim Jin Mo Kang YunJae Jung Seung Eun Baek Yun Hak Kim 《International journal of biological sciences》2022,18(13):5154
Vascular smooth muscle cell (VSMC) proliferation is a hallmark of neointimal hyperplasia (NIH) in atherosclerosis and restenosis post-balloon angioplasty and stent insertion. Although numerous cytotoxic and cytostatic therapeutics have been developed to reduce NIH, it is improbable that a multifactorial disease can be successfully treated by focusing on a preconceived hypothesis. We, therefore, aimed to identify key molecules involved in NIH via a hypothesis-free approach. We analyzed four datasets (, GSE28829, GSE43292, and GSE100927), evaluated differentially expressed genes (DEGs) in wire-injured femoral arteries of mice, and determined their association with VSMC proliferation in vitro. Moreover, we performed RNA sequencing on platelet-derived growth factor (PDGF)-stimulated human VSMCs (hVSMCs) post-phosphoenolpyruvate carboxykinase 2 (PCK2) knockdown and investigated pathways associated with PCK2. Finally, we assessed NIH formation in Pck2 knockout (KO) mice by wire injury and identified PCK2 expression in human femoral artery atheroma. Among six DEGs, only PCK2 and RGS1 showed identical expression patterns between wire-injured femoral arteries of mice and gene expression datasets. PDGF-induced VSMC proliferation was attenuated when hVSMCs were transfected with PCK2 siRNA. RNA sequencing of PCK2 siRNA-treated hVSMCs revealed the involvement of the Akt-FoxO-PCK2 pathway in VSMC proliferation via Akt2, Akt3, FoxO1, and FoxO3. Additionally, NIH was attenuated in the wire-injured femoral artery of Pck2-KO mice and PCK2 was expressed in human femoral atheroma. PCK2 regulates VSMC proliferation in response to vascular injury via the Akt-FoxO-PCK2 pathway. Targeting PCK2, a downstream signaling mediator of VSMC proliferation, may be a novel therapeutic approach to modulate VSMC proliferation in atherosclerosis. GSE120521相似文献