蝎毒耐热蛋白对大鼠急性分离海马神经元兴奋性的影响

应用全细胞膜片钳记录技术在电流钳模式下观察经持续高温等特殊处理后分离纯化的30～50 kDa蝎毒耐热蛋白(scorpion venom heat resistant protein,SVHRP)(国家发明专利,专利号ZL01 106166.92)对急性分离大鼠海马神经元兴奋性的影响.结果发现SVHRP可致海马神经元兴奋性降低.神经元经1×10-2 μg/mL SVHRP处理后动作电位发放模式改变,发放频率减少.在52个受检细胞中,有45个细胞产生位相放电(占86.54%);7个细胞产生重复放电(占13.46%).在产生位相放电的45个细胞中,有8个细胞在SVHRP处理后仍可以诱发出位相放电(占17.78%);37个细胞在SVHRP处理后无法诱导出位相放电(占82.22%),SVHRP处理后动作电位的产生与处理前相比,有显著差异(P＜0.01,n=45);在产生重复放电的7个细胞中,在1×10-2μg/mL SVHRP作用后均不能再次诱发出重复放电,而是产生一个动作电位或不再产生动作电位,药物处理前产生的动作电位个数为14.57±1.00,SVHRP处理后产生动作电位的个数为0.57±0.20,二者之间有显著性差异(P＜0.01,n=7).1×10-4 μg/mLSVHRP处理后,诱发动作电位产生的基强度由(75.10±8.99)pA增加到(119.85±12.73)pA(P＜0.01,n=8);阈电位由(-41.17±2.15)mV升至(-32.40±1.48)mV(P＜0.01,n=8);动作电位峰值由(68.49±2.33)mV下降至(54.71±0.81)mV(P＜0.01,n=8).由于神经元超兴奋性被认为是癫痫发作的基本机制之一,因此上述结果表明SVHRP有可能通过降低海马神经元兴奋性发挥其抗癫痫作用,这为蝎毒药物的进一步开发提供理论依据.     

Glutathione prevents high glucose-induced pancreatic fibrosis by suppressing pancreatic stellate cell activation via the ROS/TGFβ/SMAD pathway

The activation of pancreatic stellate cells (PSCs) is the key mechanism of pancreatic fibrosis, which can lead to β-cell failure. Oxidative stress is an important risk factor for PSC activation. There is no direct evidence proving if administration of glutathione can inhibit fibrosis and β-cell failure. To explore the role of glutathione in pancreatic fibrosis and β-cell failure induced by hyperglycaemia, we established a rat model of pancreatic fibrosis and β-cell failure. The model was founded through long-term oscillating glucose (LOsG) intake and the setup of a sham group and a glutathione intervention group. In vitro, rat PSCs were treated with low glucose, high glucose, or high glucose plus glutathione to explore the mechanism of high glucose-induced PSC activation and the downstream effects of glutathione. Compared with sham rats, LOsG-treated rats had higher reactive oxygen species (ROS) levels in peripheral leukocytes and pancreatic tissue while TGFβ signalling was upregulated. In addition, as the number of PSCs and pancreatic fibrosis increased, β-cell function was significantly impaired. Glutathione evidently inhibited the upregulation of TGFβ signalling and several unfavourable outcomes caused by LOsG. In vitro treatment of high glucose for 72 h resulted in higher ROS accumulation and potentiated TGFβ pathway activation in PSCs. PSCs showed myofibroblast phenotype transformation with upregulation of α-SMA expression and increased cell proliferation and migration. Treatment with either glutathione or TGFβ pathway inhibitors alleviated these changes. Together, our findings suggest that glutathione can inhibit PSC activation-induced pancreatic fibrosis via blocking ROS/TGFβ/SMAD signalling in vivo and in vitro.Subject terms: Pre-diabetes, Pre-diabetes     

北京东灵山森林植物多样性的网络结构特征

一个群落可以看作是由物种相互连接的复杂系统,刻画其网络结构有助于深入揭示系统性质以及结构与功能之间的关系。在生态学中,复杂网络理论已被成功应用于食物网与互利网络的结构研究,但尚未检验其刻画生物多样性格局的能力。采用复杂网络理论研究了北京东灵山森林乔木层、灌木层、草本层植物关联关系的网络结构特征及其差异。结果表明,植物物种的共同出现是非随机的,并表现出一定的小世界模式;乔木层、灌木层、草本层植物共同出现的网络在结构特征上存在明显差异,草本层网络比乔木层和灌木层网络更松散且平均路径更长,灌木层和草本层网络的聚类系数高于乔木层且存在度的幂律分布。皆表明复杂网络理论具备反映不同层植物多样性格局差异的能力。     

A Fluorescent Recombinase Aided Amplification Assay for Detection of Babesia microti

Babesia microti is one of the most common causative agents of babesiosis. A sensitive and rapid detection is necessary for screening potentially infected individuals. In this study, B. microti cytochrome c oxidase subunit I (cox1) was selected as the target gene, multiple primers were designed, and optimized by a recombinase-aided amplification (RAA) assay. The optimal primers and probe were labeled with fluorescein. The sensitivity of fluorescent RAA (fRAA) was evaluated using gradient diluents of the cox1 recombinant plasmid and genomic DNA extracted from whole blood of B. microti infected mice. The specificity of fRAA was assessed by other transfusion transmitted parasites. The analytical sensitivity of the fRAA assay was 10 copies of recombinant plasmid per reaction and 10 fg/μl B. microti genomic DNA. No cross-reaction with any other blood-transmitted parasites was observed. Our results demonstrated that the fRAA assay would be rapid, sensitive, and specific for the detection of B. microti.     

Direct adenylation from 5′-OH-terminated oligonucleotides by a fusion enzyme containing Pfu RNA ligase and T4 polynucleotide kinase

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.     

气温和根区温度对葡萄叶片光合荧光特性的影响

为探究气温和根区温度对葡萄(Vitis vinifera)叶片光合荧光特性的影响,以一年生巨峰葡萄为试材,设置对照、高气温、高根区温度和两者交叉作用共4组处理。结果表明,相较于对照和高气温,高根区温度以及交叉处理叶片最大光化学效率(Fv/Fm)降低更明显;与对照相比,高根区温度以及高气温与高根区温度交叉处理下光系统II (PSII)实际光化学效率Y(II)显著降低,非调节能量耗散的量子产量Y(NPQ)及QA氧化还原状态(1–qP)值显著上升。同时,高根区温度以及高气温与高根区温度交叉处理显著增加了J点的可变荧光(Vj),而用于电子传递的量子产额(φEo)及性能指数(PIABS)显著降低。此外,高根区温度以及高气温与高根区温度交叉处理下单位面积有活性的反应中心数目(RC/CSm)也显著下降,K点相对可变荧光(Wk)明显上升。综上所述,高根区温度是高气温与根区高温交叉胁迫的主导因子, PSII受体侧是主要的伤害位点,高气温加剧了高根区温度对PSII造成的伤害。     

Targeting ACLY efficiently inhibits SARS-CoV-2 replication

The Coronavirus Disease 2019 (COVID-19) pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the biggest public health challenge the world has witnessed in the past decades. SARS-CoV-2 undergoes constant mutations and new variants of concerns (VOCs) with altered transmissibility, virulence, and/or susceptibility to vaccines and therapeutics continue to emerge. Detailed analysis of host factors involved in virus replication may help to identify novel treatment targets. In this study, we dissected the metabolome derived from COVID-19 patients to identify key host factors that are required for efficient SARS-CoV-2 replication. Through a series of metabolomic analyses, in vitro, and in vivo investigations, we identified ATP citrate lyase (ACLY) as a novel host factor required for efficient replication of SARS-CoV-2 wild-type and variants, including Omicron. ACLY should be further explored as a novel intervention target for COVID-19.     

北京东灵山土壤动物-植物互作关系对海拔格局的响应

土壤动物可能会与植物形成复杂的互作网络,有关其对海拔梯度做出响应的研究还相对较少,了解不同生境下的生态网络为理解和预测系统的稳定性及其动态变化机制提供了新途径。于北京东灵山地区调查了沿海拔梯度凋落物层与土壤层中的土壤动物以及木本和草本植物的多样性,并通过零模型的方法对比了土壤动物-植物互作网络特征参数的标准化效应值在高-中-低三个海拔段的差异。结果显示：土壤动物的稀有类群在中海拔段多样性较高,木本植物多样性随海拔降低,而草本植物多样性则随海拔升高。各生物类群组成都随海拔距离而增加,存在明显的聚集性分布格局。植物与土壤动物之间的互作网络特征参数不仅随海拔改变,且在凋落物与土壤层之间、木本与草本植物之间存在差异。总体而言,土壤动物与木本植物互作网络的海拔变化主要与土壤动物多样性有关,而与草本的互作网络则与草本植物多样性有关。沿海拔梯度,存在土壤动物-木本植物向土壤动物-草本植物网络的适应性转变。本研究在同一海拔梯度上探讨了不同层土壤动物与植物的互作关系,对深入理解山地生态系统中各生物类群的共存机制具有重要意义。     

辛伐他汀对动脉粥样硬化大鼠凋亡相关基因Fas及FasL蛋白表达的影响

目的通过研究辛伐他汀对动脉粥样硬化大鼠血管壁中细胞凋亡相关基因Fas及FasL蛋白表达产物的影响,探讨其在预防动脉粥样硬化发生中的可能机制。方法复制动脉粥样硬化大鼠模型,以辛伐他汀干预,取胸主动脉,观察其斑块变化,采用免疫组化Elivision法测定动脉粥样硬化血管壁中Fas、FasL蛋白表达。结果Fas蛋白表达在实验组明显高于对照组及干预组(P<0.01,P<0.05),实验组FasL蛋白表达也明显高于对照组及干预组(P<0.05)。结论Fas及FasL基因通过促进细胞凋亡作用而诱发动脉粥样硬化过程,辛伐他汀可通过调节细胞凋亡过程发挥抗动脉粥样硬化作用。     

锯缘青蟹精子发生的超微结构

采用透射电镜观察锯缘青蟹精子发生过程中超微结构的变化,结果表明：精原细胞椭圆形,染色质分布于核膜周围,胞质中具嵴少的线粒体,内质网小泡等。初级精母细胞染色质呈非浓缩状,胞质中具众 内质网小泡,特殊的膜系及晶格状结构。次级精母细胞核质间出现由内质小泡聚集成的腔。