Thermococcus sp.B1001环糊精酶的重组表达及在麦芽七糖制备中的应用

成功构建pET-24a-tscd质粒,实现Thermococcus sp.Strain B1001来源的环糊精酶(TsCDase)在Es-cherichia coli BL21(DE3)中表达.通过热处理和镍柱分离对重组TsCDase进行纯化.酶学性质研究表明,重组TsCDase的比活为1 208.04 U/mg,最适温度为90℃、最适pH值为5.5.重组酶TsCDase在85℃、90℃、95℃条件下的半衰期分别为180、120、30min.酶转化研究表明,以80 g/L β-环糊精为底物,当酶转化温度为90℃、反应pH值为5.5～6.0,加酶量为25 U/g,反应时间为4 h时,麦芽七糖产率为81.19％和85.95％,七糖占产物麦芽寡糖的比例为95.24％和92.92％.本研究结果为工业化制备麦芽七糖奠定良好基础.     

基于mtDNA D-loop分析高原鳅群体遗传多样性和系统发育

本研究以高原鳅属6个群体共82尾个体的线粒体DNA D-loop高变区为标记,采用分子生物信息学方法分析其遗传多样性及系统发育关系.结果表明,在570 bp的mtDNA D-loop中共检测到149个多态位点,由此界定了 39个单倍型.在6个群体中,单倍型多样度(Hd)由高到低依次为细尾高原鳅(T.stenura)(0.899)＞东方高原鳅(T.orientalis)＞似鲶高原鳅(T.siluroides)＞达里湖高原鳅(T.dalaica)/黄河高原鳅(T.pap-penheimi)＞湘西盲高原鳅(T.xiangxiensis)(0.286);核苷酸多样度(Pi)由高到低依次为似鲶高原鳅(0.042 59)＞黄河高原鳅＞细尾高原鳅＞东方高原鳅＞达里湖高原鳅＞湘西盲高原鳅(0.000 5);细尾高原鳅与湘西盲高原鳅群体间遗传距离(0.186)最大,达里湖高原鳅与东方高原鳅群体间遗传距离(0.027)最小;细尾高原鳅与达里湖高原鳅群体间遗传分化程度(0.989)最大,似鲶高原鳅与东方高原鳅群体间遗传分化程度(0.222)最小.其中仅有一个单倍型(Hap29)为达里湖高原鳅与似鲶高原鳅群体所共有,其余均为特有单倍型.进化树显示6个高原鳅群体分为三大支,其中细尾高原鳅自成一支;湘西盲高原鳅与黄河高原鳅聚成一支;其余东方高原鳅、达里湖高原鳅和似鲶高原鳅聚成一支.Network与系统发育树结果基本一致.就单倍型丰富度和核苷酸多样性而言,似鲶高原鳅遗传多样性较高,湘西盲高原鳅遗传多样性较低.就系统发育而言,群体间的分化程度支持了其分类地位;似鲶高原鳅、东方高原鳅和达里湖高原鳅的亲缘关系相对较近;不同的聚类推测高原鳅可能至少经历了 3次驯化事件.本研究结果为高原鳅鱼类生物多样性保护和评价提供遗传学理论依据.     

Evaluation of the efficacy and safety of hydroxychloroquine in comparison with chloroquine in moderate and severe patients with COVID-19

Science China Life Sciences -     

Experimental warming shifts coupling of carbon and nitrogen cycles in an alpine meadow

增温对高寒草甸生态系统碳氮循环耦合关系的影响 陆地生态系统碳吸收受土壤氮素可用性的调节。然而,全球变化背景下的不同生态系统组分的碳氮比及其所反映的碳氮循环耦合关系尚不十分清楚。本文运用数据同化的方法,将一个高寒草甸增温试验的14组数据同化到草地生态系统模型中,从而评估了增温如何影响陆地生态系统的碳氮循环耦合关系。研究结果表明,增温提高了土壤氮素的有效性,降低了土壤活性碳库的碳氮比,导致植物对土壤氮的吸收增加。但是由于植物叶片吸收的碳比吸收的氮增加更多,使得叶片中碳氮比增加,而根部的碳输入增加则低于氮的增加,导致根部的碳氮比减少。同时,增温降低了凋落物碳氮比,可能是在土壤高氮有效性的条件下,凋落物氮的固定得到增强;而且增温加速了凋落物的分解。同时增温还增加了慢速土壤有机质的碳氮比,使得该土壤碳库的碳固存潜力增大。由于大多数模型在不同的环境中通常使用相对固定的碳氮比,本研究所发现的气候变暖条件下碳氮比的差异变化可为模型参数化提供一个有效的参考,有利于模型对未来气候变化背景下生态系统碳氮耦合关系响应的预测。     

LncRNA-HOTAIR promotes endothelial cell pyroptosis by regulating the miR-22/NLRP3 axis in hyperuricaemia

Long non-coding RNA (lncRNA) plays an important role in the renal inflammatory response caused by hyperuricaemia. However, the underlying molecular mechanisms through which lncRNA is involved in endothelial injury induced by hyperuricaemia remain unclear. In this study, we investigated the regulatory role of lncRNA-HOTAIR in high concentration of uric acid (HUA)–induced renal injury. We established hyperuricaemia mouse model and an in vitro uric acid (UA)–induced human umbilical vein endothelial cell (HUVEC) injury model. In HUA-treated HUVECs and hyperuricaemia mice, we observed increased HOTAIR and decreased miR-22 expression. The expression of pyroptosis-associated protein (NLRP3, Caspase-1, GSDMD-N, GSDMD-FL) was increased. The release of LDH, IL-1β and IL-18 in cell supernatants and the sera of model mice was also increased. The proliferation of HUVECs stimulated by HUA was significantly inhibited, and the number of TUNEL-positive cells in hyperuricaemia mouse kidney was increased. Bioinformatics analysis and luciferase reporter and RIP assays confirmed that HOTAIR promoted NLRP3 inflammasome activation by competitively binding miR-22. In gain- or loss-of-function experiments, we found that HOTAIR and NLRP3 overexpression or miR-22 knock down activated the NLRP3 inflammasome and promoted pyroptosis in HUA-treated HUVECs, while NLRP3 and HOTAIR knockdown or a miR-22 mimic exerted the opposite effects. Furthermore, in vivo experiments validated that HOTAIR knockdown alleviated renal inflammation in hyperuricaemia mice. In conclusion, we demonstrated that in hyperuricaemia, lncRNA-HOTAIR promotes endothelial cell pyroptosis by competitively binding miR-22 to regulate NLRP3 expression.     

中国脊椎动物2020年新增物种

为了及时掌握中国脊椎动物新增物种情况, 本文系统检索和整理了2020年发表的分类学文献, 汇总结果表明: 2020年中国脊椎动物共新增109种, 其中新物种100种, 国家级新记录种9种。包括鱼类新种24种、两栖类新种41种和新记录4种、爬行类新种30种和新记录4种、鸟类新种1种、哺乳类新种4种和新记录1种。上述新增物种中有92种描述报道时应用了分子遗传学证据, 占新增物种数量的84.4%。在新增脊椎动物物种中, 两栖类集中于无尾目、爬行类全部属于有鳞目, 分别有43种和34种, 其累计占比超新增物种总数的70%。在云南、西藏、湖南、贵州、四川等省区发现新物种数量较多, 均有10种及以上, 累计占比超新增物种总数的60%。绝大部分物种为中国学者发表, 绝大多数论文发表于英文期刊。总结数据提示今后需持续加强我国低等脊椎动物多样性的调查研究, 重视运用分子系统学技术进行物种识别。     

2010-2020年中华穿山甲在中国的发现记录及保护现状

中国是中华穿山甲(Manis pentadactyla)历史分布区面积和野生种群数量最大的国家。中华穿山甲曾广泛见于我国长江以南各省, 但20世纪中期以来, 由于其甲片被作为贵重的中药材原料, 加之地下野味市场的需求, 大量非法捕猎使得我国野生穿山甲经历了剧烈的分布区缩减和种群下降。目前, 中华穿山甲已被列为我国一级重点保护野生动物, 在IUCN红色名录中被评估为极危(CR)等级。中华穿山甲分布范围广、种群密度低、活动隐秘、调查难度大, 摸清其野外分布现状是当前穿山甲研究与保护中的首要任务。为此, 本研究通过检索2010-2020年间全国范围内中华穿山甲的发现记录, 统计中华穿山甲的目击数量、空间分布以及后续状态, 制作物种分布地图, 并与历史分布情况进行对比, 以评估其野生种群的分布现状与分布区变化。2010-2020年共在11个省级行政区收集到中华穿山甲确认记录142条, 主要集中于大陆华东地区及台湾岛, 台湾、浙江、广东三省记录位点数占全部位点数的67.6%; 相较其历史分布区, 西南、华南地区近年来野外确认较为匮乏。近10年来我国的中华穿山甲记录呈逐渐增多趋势, 86%被发现的实体穿山甲得到救助、放归或未被干扰。本研究结果表明, 目前中华穿山甲在我国, 尤其是华东及台湾地区, 仍具有一定数量的野生种群分布, 且近年来民众对于穿山甲的认知和保护意识有了较大提高。 然而, 现有调查和资料仍不足以对该物种进行全面的现状评估, 亟需加强对我国野生穿山甲种群的调查、监测和保护。     

Procyanidin B1 promotes in vitro maturation of pig oocytes by reducing oxidative stress

Oxidative stress negatively affects the in vitro maturation (IVM) of oocytes. Procyanidin B1 (PB1) is a natural polyphenolic compound that has antioxidant properties. In this study, we investigated the effect of PB1 supplementation during IVM of porcine oocytes. Treatment with 100 μM PB1 significantly increased the MII oocytes rate (p <0.05), the parthenogenetic (PA) blastocyst rate (p <0.01) and the total cell number in the PA blastocyst (p < 0.01) which were cultured in regular in vitro culture (IVC) medium. The PA blastocyst rate of regular MII oocytes activated and cultured in IVC medium supplemented with 100 and 150 μM PB1 significantly increased compared with control (p < 0.01 and p < 0.05). We also evaluated the reactive oxygen species (ROS) levels, mitochondrial membrane potential (Δψm) levels, glutathione (GSH) levels, and apoptotic levels in MII oocytes and cumulus cells following 100 μM PB1 treatment. The results showed that the PB1 supplementation decreased ROS production and apoptotic levels. In addition, PB1 was found to increase Δψm levels and GSH levels. In conclusion, PB1 inhibited apoptosis of oocytes and cumulus cells by reducing oxidative stress. Moreover, PB1 improved the quality of oocytes and promoted PA embryo development. Taken together, our results suggest that PB1 is a promising antioxidant additive for IVM of oocytes.