可降解新烟碱类杀虫剂微生物及其代谢途径的研究进展

新烟碱类化合物基于烟碱结构改造修饰制备,相较菊酯、含磷类等杀虫剂,因其选择性毒力被认为是一类对人类和生态无害的农药。然而,近年来由于新烟碱类杀虫剂(neonicotinoid insecticides)过度施用,其残余或转化的物质通过在土壤与水体中累积,影响昆虫甚至哺乳动物及其生理与行为,导致了一系列生态环境问题和继发危害。本文聚焦新烟碱类杀虫剂的产业现状,面向生物降解新烟碱类杀虫剂这一迫切需求,围绕新烟碱类杀虫剂的微生物菌株资源,重点阐述微生物降解新烟碱类杀虫剂的代谢机制及其多样性。通过梳理新烟碱类杀虫剂生物降解及其应用转化的关键问题和前沿进展,旨在为借助合成生物学和宏基因组学手段建立或筛选安全可控的新烟碱类杀虫剂的高效转化体系提供参考。     

基于转录组学的白扁豆总皂苷抑制前列腺癌细胞系PC-3细胞生长的机制研究

目的 白扁豆总皂苷是中药白扁豆经过提取分离纯化步骤制备得到,关于白扁豆的总皂苷成分如何影响前列腺癌细胞系PC-3细胞的生长情况缺少研究。因此,有必要探讨白扁豆总皂苷对前列腺癌细胞系PC-3细胞生长的机制研究。方法 本研究采用CCK8方法检测不同浓度的白扁豆总皂苷对前列腺癌细胞系PC-3细胞生长的影响。利用转录组学分析白扁豆总皂苷抑制前列腺癌细胞系PC-3细胞生长的分子机制,并且进一步通过实时定量PCR（qRT-PCR）和蛋白质免疫印迹（Western blot）实验对相关差异基因的表达进行验证。利用Western blot和CCK8检测白扁豆总皂苷处理过表达醛脱氢酶7家族成员A1（ALDH7A1）的PC-3细胞存活率。结果 随着白扁豆总皂苷浓度升高,前列腺癌细胞PC-3的存活率显著下降,白扁豆总皂苷的IC₅₀值为1 086 mg/L。转录组学测序结果显示,与对照组相比,白扁豆总皂苷处理的细胞中有2 360个差异表达基因,其中1 982个基因上调,378个基因下调。基因功能注释（GO）结果显示,差异表达基因显著富集到与有丝分裂纺锤体检查点（mitotic spindle checkpoint）、纺锤体组装检查点（spindle assembly checkpoint）等一系列跟癌症的发生发展密切相关的生物学过程。此外,基因组京都百科全书（KEGG）分析结果也显示,差异表达基因富集在肿瘤代谢等信号通路。进一步对其中的差异基因进行验证,结果显示,与对照组相比,白扁豆总皂苷处理的前列腺癌细胞中ALDH7A1、甘氨酸C-乙酰转移酶（GCAT）和磷酸甘油酸变位酶家族成员4（PGAM4）的蛋白质表达水平明显降低（P<0.05）,而二甲基甘氨酸脱氢酶（DMGDH）和胱硫醚β合成酶样（CBSL）的蛋白质表达水平显著升高（P<0.001）。体外细胞实验结果表明,白扁豆总皂苷通过下调前列腺癌细胞中ALDH7A1的表达抑制PC-3细胞生长。结论 白扁豆总皂苷可能通过下调ALDH7A1表达从而在体外抑制前列腺癌细胞的生长。     

植物苯丙烷代谢途径

众所周知,固着生长的植物经常受到环境中各种生物和非生物胁迫的威胁。所以在漫长的进化过程中,植物必须将多样的环境信号整合到其发育过程中,以实现适应性形态的发生和代谢途径的精确调控,最终使植物完成整个生长周期。研究显示,苯丙烷代谢作为植物重要的次级代谢途径之一,其代谢产物,例如木质素、孢粉素、花青素和有机酸等,在调控植物适应性生长的过程中发挥着重要功能。特别是在药用植物中,苯丙烷代谢还与众多药用活性成分的合成息息相关,几乎所有包含苯丙烷骨架的天然药效成分均由苯丙烷代谢途径直接或间接合成,例如黄酮类、萜类和酚类等。此外,经苯丙烷代谢途径产生的一些次级代谢产物还能由植物根系外泌到周际土壤中,通过改变根系微生物的菌群生态,而影响植物生长和抵抗生物或非生物胁迫的能力。同时,苯丙烷代谢介导的这种植物-微生物互作也与药用植物的道地品质密不可分。本文综述了近年来植物苯丙烷代谢途径的最新研究进展,重点对该代谢途径中代谢产物的生理功能及表达调控机制进行了介绍,以期更深入地理解药用植物苯丙烷代谢与药材性状之间的潜在关系,旨在指导优良中草药的遗传育种,以进一步促进我国中医药事业的蓬勃发展。     

土壤湿度对香樟幼苗根系生长和构型的影响

以1年生香樟(Cinnamomum camphora)幼苗为试材,设置对照组(CK)、中度干旱处理(M)、重度干旱处理(S)三个处理,比较不同土壤湿度下香樟幼苗不同时期地上部分生长和根系构型,探究香樟幼苗根系对不同土壤湿度的适应性及其耐旱机制。结果表明,中度和重度干旱处理组的香樟根系及地上部分干物质积累、根系长度、根系表面积、根系直径和根尖数均显著小于对照组(P<0.05)。同时干旱显著增加香樟幼苗的根系拓扑指数,降低香樟根的分形维数和平均分枝角度(P<0.05)。可见土壤湿度程度及处理时间显著影响香樟根系的生长及在土壤中的布局。较低土壤湿度可显著抑制根长的延长、根表面积扩大和根的增殖,且随着土壤湿度的继续降低以及处理时间延长,香樟幼苗根系的生长受到水分亏缺的抑制作用加重,根系建成成本增高的同时,根系分枝的复杂性降低,根系必须通过朝着更陡、更深的方向生长伸长来提高水分吸收效率。建议在园林绿化工程养护过程中制定科学的水分管理策略,以满足香樟生长过程中对土壤水分的需要。     

Neutralization of five SARS-CoV-2 variants of concern by convalescent and BBIBP-CorV vaccinee serum

The prevalence of SARS-CoV-2 variants of concern (VOCs) is still escalating throughout the world. However, the level of neutralization of the inactivated viral vaccine recipients’ sera and convalescent sera against all VOCs, including B.1.1.7 (Alpha), B.1.351 (Beta), P.1 (Gamma), B.1.617.2 (Delta), and B.1.1.529 (Omicron) remains to be lack of comparative analysis. Therefore, we constructed pseudoviruses of five VOCs using a lentiviral-based system and analyzed their viral infectivity and neutralization resistance to convalescent and BBIBP-CorV vaccinee serum at different times. Our results show that, compared with the wild-type strain (WT), five VOC pseudoviruses showed higher infection, of which B.1.617.2 and B.1.1.529 variant pseudoviruses exhibited higher infection rates than wild-type or other VOC strains, respectively. Sera from 10 vaccinated individuals at the 1, 3 and 5-month post second dose or from 10 convalescent at 14 and 200 days after discharge retained neutralizing activity against all strains but exhibited decreased neutralization activity significantly against the five VOC variant pseudoviruses over time compared to WT. Notably, 100% (30/30) of the vaccinee serum samples showed more than a 2.5-fold reduction in neutralizing activity against B.1.1.529, and 90% (18/20) of the convalescent serum samples showed more than 2.5-fold reduction in neutralization against B.1.1.529. These findings demonstrate the reduced protection against the VOCs in vaccinated and convalescent individuals over time, indicating that it is necessary to have a booster shot and develop new vaccines capable of eliciting broad neutralization antibodies.     

CSN5 promotes the invasion and metastasis of pancreatic cancer by stabilization of FOXM1

COP9 signalosome subunit 5 (CSN5) has been involved in the progression of diverse human cancers. MMP2 plays an important role in the metastasis of cancer cells. However, the roles and relationship of in pancreatic cancer (PC) is still unknown. Here, our data shown that both CSN5 and MMP2 were significantly upregulated in PC compared with the corresponding adjacent tissues, where a positive correlation in their expression and associated malignant characteristics were found. Further, silencing of CSN5 expression markedly inhibited PC invasion and metastasis in vitro and in vivo, accompanied by decreased MMP2 expression. Moreover, the anti-metastasis role of CSN5 silence was reversed by MMP2 overexpression, whereas knockdown of MMP2 decreased PC metastasis driven by upregulation of CSN5. Further investigation revealed that CSN5 regulated MMP2 expression via activation of FOXM1 in PC cells. Mechanistically, CSN5 directly bound FOXM1 and decreased its ubiquitination to enhance the protein stability of FOXM1. Taken together, the results indicate that CSN5 can contribute to PC invasion and metastasis through activation of FOXM1/MMP2 axis.     

Hypoxic conditioned medium derived from bone marrow mesenchymal stromal cells protects against ischemic stroke in rats

In recent years, studies have shown that the secretome of bone marrow mesenchymal stromal cells (BMSCs) contains many growth factors, cytokines, and antioxidants, which may provide novel approaches to treat ischemic diseases. Furthermore, the secretome may be modulated by hypoxic preconditioning. We hypothesized that conditioned medium (CM) derived from BMSCs plays a crucial role in reducing tissue damage and improving neurological recovery after ischemic stroke and that hypoxic preconditioning of BMSCs robustly improves these activities. Rats were subjected to ischemic stroke by middle cerebral artery occlusion and then intravenously administered hypoxic CM, normoxic CM, or Dulbecco modified Eagle medium (DMEM, control). Cytokine antibody arrays and label-free quantitative proteomics analysis were used to compare the differences between hypoxic CM and normoxic CM. Injection of normoxic CM significantly reduced the infarct area and improved neurological recovery after stroke compared with administering DMEM. These outcomes may be associated with the attenuation of apoptosis and promotion of angiogenesis. Hypoxic preconditioning significantly enhanced these therapeutic effects. Fourteen proteins were significantly increased in hypoxic CM compared with normoxic CM as measured by cytokine arrays. The label-free quantitative proteomics analysis revealed 163 proteins that were differentially expressed between the two groups, including 107 upregulated proteins and 56 downregulated proteins. Collectively, our results demonstrate that hypoxic CM protected brain tissue from ischemic injury and promoted functional recovery after stroke in rats and that hypoxic CM may be the basis of a potential therapy for stroke patients.