程序性细胞死亡因子10抑制Ⅰ型干扰素表达并促进FMDV复制

旨在探究宿主蛋白程序性细胞死亡因子10（programmed cell death factor 10，PDCD10）通过抑制Ⅰ型干扰素表达进而促进口蹄疫病毒（foot-and-mouth disease virus，FMDV）的复制。首先，本研究验证了过表达和沉默PDCD10对FMDV复制的影响，接着利用双荧光素酶报告系统探究PDCD10对Ⅰ型干扰素信号通路活化的影响，最后，利用实时荧光定量PCR探究PDCD10对Ⅰ型干扰素通路下游刺激基因（IFN-stimulated genes，ISGs）转录的影响。结果表明，过表达PDCD10显著促进FMDV的复制，沉默PDCD10显著抑制FMDV的复制。与对照相比，过表达PDCD10后感染仙台病毒（Sendai virus，SeV）的细胞培养液上清液显著促进FMDV复制，进一步，PDCD10显著抑制SeV诱导的IFN-β启动子以及NF-κB的激活且呈剂量依赖性，并且PDCD10负调控Ⅰ型干扰素通路信号分子转录，最后还发现PDCD10负调控Ⅰ型干扰素下游ISGs转录。本研究结果为深入探究PDCD10在抗病毒天然免疫中的作用积累了资料。     

凤阳山不同林分土壤腐殖质特征及影响因素

以浙江省凤阳山海拔1 300~1 400 m处不同林分类型(阔叶混交林、针阔混交林、杉木林、竹林)为对象,测定不同土层土壤基本理化性质、酶活性及腐殖质质量分数,分析土壤腐殖质特征及影响因素,为凤阳山自然保护区的土壤肥力和可持续经营发展提供理论依据。结果表明:土壤腐殖质质量分数针阔混交林最高,阔叶混交林、竹林、杉木林次之;胡敏酸质量分数针阔混交林最高,杉木林最低;富里酸质量分数针阔混交林最高,竹林最低。除磷酸酶活性随土层加深无统一规律外,4种林分脲酶活性、过氧化氢酶活性及蔗糖酶活性土壤随土层加深皆呈现降低趋势。土壤pH值、土壤密度与土壤腐殖质质量分数、胡敏酸质量分数和富里酸质量分数均呈现显著的负相关;除磷酸酶活性相关性不显著外,土壤脲酶活性、蔗糖酶活性、过氧化氢酶活性、总孔隙度、毛管孔隙、非毛管孔隙与土壤腐殖质质量分数、胡敏酸质量分数和富里酸质量分数均呈现显著的正相关。凤阳山不同林分类型对土壤腐殖质特征的影响较土层深度更显著,土壤理化性质及酶活性与土壤腐殖质质量分数有着密切关系。     

不同施肥处理对马铃薯农田土壤理化性状及产量的影响

为了探究不同施肥处理对马铃薯农田土壤理化性状及产量的影响,试验地采用马铃薯连作模式,设置3个施肥处理,即：单施化肥(T)、有机肥配施化肥(YTF 1/2)、全量有机肥(YTF)。结果表明：在不同施肥处理下马铃薯农田土壤的理化性质和马铃薯产量发生了变化,其中变化最为明显的土壤指标有土壤容重、孔隙度、饱和导水率、有效磷。YTF处理较T处理可分别显著(P<0.05)降低土壤容重16.8%,增加土壤孔隙度12.7%,提升饱和导水率25.3%。YTF处理可显著提升土壤有效磷含量43.0%,但各处理间土壤pH、有机碳、全氮、全磷、碱解氮、速效钾之间差异并未达到显著水平。同时,较之T处理,YTF处理亦可显著提升土壤团聚体含量。YTF和YTF 1/2处理可分别较T处理提升马铃薯产量24.6%和12.8%。因此,施用有机肥不仅可以改善土壤结构,改良土壤物理性状,亦能促产增收。     

miR-140-3p inhibits viability,invasion and migration of non-small-cell lung cancer A549 cells by targeting PD-L1

AIM: To explore the target relationship between microRNA-140-3p (miR-140-3p) and programmed cell death ligand 1 (PD-L1) and their effect on the viability, migration and invasion of non-small-cell lung cancer A549 cells.METHODS: RT-qPCR was used to detect the miR-140-3p expression in HLF-1, A549 and H1299 cells, and then the A549 cells with the most significant difference were selected as the subsequent research object. TargetScan software and dual-luciferase reporter assay were performed to predict and confirm the target relationship between miR-140-3p and PD-L1. RT-qPCR and Western blot were used to determine the effects of miR-140-3p mimic and inhibitor on PD-L1 expression level. MTT assay was used to detect the viability of A549 cells. Transwell assay was performed to detect the migration and invasion abilities of the A549 cells.RESULTS: miR-140-3p was significantly down-regulated in the A549 cells and H1299 cells (P<0.05). Transfection with miR-140-3p mimic decreased the expression of PD-L1 and inhibited the viability, migration and invasion of the A549 cells. Transfection with pcDNA3.0-PD-L1 reversed the inhibitory effect of miR-140-3p on the viability, migration and invasion of the A549 cells.CONCLUSION: miR-140-3p inhibits the viability, migration and invasion of A549 cells by targeting PD-L1.     

种植年限对设施蔬菜土壤养分和环境的影响

为了给设施蔬菜地合理施肥提供依据,研究了不同种植年限设施蔬菜地土壤理化性质、微生物数量及酶活性变化,以露地土壤为对照(CK),采集种植1 a(年)、3 a、5 a、7 a、9 a、11 a、13 a、15 a、17 a、20 a以及20 a以上的0~20 cm设施蔬菜土壤样品,采用常规土壤理化性质检测方法测定了土壤理化性质和酶活性,采用平板计数法,分析土壤微生物数量。结果表明,设施菜地种植年限与土壤容重呈正相关,与土壤孔隙度呈负相关;设施菜地土壤pH值随种植年限的增加逐年降低,两者呈显著的负相关性;土壤EC值、土壤有机质和速效磷含量都随种植年限的增加而逐年增高,并与种植年限呈显著的正相关性;与CK相比,种植13 a的蔬菜样地土壤全氮和速效钾含量均显著地增加;连续种植20 a以上,设施蔬菜土壤含盐量比对照土壤显著地增加了486.49%。土壤细菌和放线菌数量、过氧化氢酶和碱性磷酸酶活性呈现出先增加后减少的趋势,且都在种植年限为11~13 a时达到最大;真菌数量随着种植年限的增加而增加,而土壤脲酶活性则随种植年限的增加呈下降趋势;转化酶活性随着种植年限的增加保持不变。土壤微生物数量与酶活性之间存在一定程度的相关关系,其中土壤真菌数量与土壤酶活性之间的相关性最为显著。综上所述,种植年限不同的设施菜地,土壤养分失衡,呈现酸化趋势,盐分含量逐年增加,土传真菌病害潜在发生,对设施蔬菜生产不利。     

内生菌与雷公藤细胞悬浮共培养的生理生化特性

将2株雷公藤内生真菌Fusarium oxysporum NS33与Penicillium steckii NS6、2株内生细菌Enterobacter cloacae sub sp LG3与Serratia marcescens LY1及其组合分别与雷公藤细胞悬浮共培养,对不同培养体系内雷公藤细胞的生长及其生理生化特征进行研究。结果显示,在共培养前期,与对照相比,接种单一内生菌株提高了细胞的干重,其中菌株NS6的促生效果最明显;而在共培养后期,无论是单一内生菌还是混合内生菌均对细胞生长具有抑制作用。内生真菌和菌株组合处理的培养液p H值有明显的升高,而内生细菌LY1则明显降低了培养液的p H值,其具有产酸性。另外,当雷公藤细胞同混合菌株共培养时,培养基中总糖消耗量是最大的,而接种单独菌株时则对细胞可溶性蛋白含量具有一定的提高作用。接种内生菌会影响雷公藤细胞的POD、CAT及SOD活性,与对照相比,接种单独菌株会更加提升POD与CAT的活性,而细胞MDA含量则明显下降。     

The M43 domain-containing metalloprotease RcMEP1 in Rhizoctonia cerealis is a pathogenicity factor during the fungus infection to wheat

Wheat(Triticum aestivum L.) is an important staple crop for global human. The necrotrophic fungus Rhizoctonia cerealis is the causal pathogen of sharp eyespot, a devastating disease of wheat. Herein, we identified RcMEP1, a zinc metalloproteaseencoding gene from R. cerealis genomic sequences, and characterized its pathogenesis function. RcMEP1 expressed at markedly-high levels during R. cerealis infection process to wheat. The predicted protein RcMEP1 comprises of 287 amino acid residues and contains a signal peptide and a M43 metalloprotease domain harboring the active site motif(HEVGHWLGLYH). The assays of Agrobacterium tumefaciens-mediated transient expression in Nicotiana benthamiana leaves indicated that RcMEP1 is an apoplastic elicitor of cell death, and that the predicted signal peptide functions and is required for secretion and cell death-induction. The purified RcMEP1 protein and its M43 domain peptide were individually able to induce plant cell death and H2 O2 accumulation, and to inhibit expression of host chitinases when infiltrated into wheat and N. benthamiana leaves, while the M43 domain-deleting peptide and negative control lacked the capacity. Moreover, compared with the control pretreatment, the purified RcMEP1 protein or its M43-domain peptide resulted in enhanced pathogenesis in the inoculated wheat, whereas the M43 domain-deleting peptide failed. These results suggest that RcMEP1 acted as an important pathogenicity factor during R. cerealis infection to wheat and that its signal peptide and M43 domain are required for the secretion and pathogenesis of RcMEP1. This study provides insights into pathogenesis role of M43 domain-containing metalloproteases during R. cerealis infection to wheat.     

高山油菜花粉黄酮提取工艺研究

采用超声波结合复合酶液对高山油菜花粉进行破壁预处理后，以95%的乙醇提取花粉中黄酮，研究提取温度、时间和固液比对破壁高山油菜花粉黄酮得率的影响，筛选适宜的提取工艺。结果表明，超声波酶解处理时间60 min即可使油菜花粉破壁率达到90%以上；通过正交试验确定最佳提取工艺条件为：提取温度95 ℃，时间2 h，料液比1∶35 (g/mL)，此时高山油菜花粉黄酮得率高达7.65%。     

EPS364, a Novel Deep-Sea Bacterial Exopolysaccharide,Inhibits Liver Cancer Cell Growth and Adhesion

The prognosis of liver cancer was inferior among tumors. New medicine treatments are urgently needed. In this study, a novel exopolysaccharide EPS364 was purified from Vibrio alginolyticus 364, which was isolated from a deep-sea cold seep of the South China Sea. Further research showed that EPS364 consisted of mannose, glucosamine, gluconic acid, galactosamine and arabinose with a molar ratio of 5:9:3.4:0.5:0.8. The relative molecular weight of EPS364 was 14.8 kDa. Our results further revealed that EPS364 was a β-linked and phosphorylated polysaccharide. Notably, EPS364 exhibited a significant antitumor activity, with inducing apoptosis, dissipation of the mitochondrial membrane potential (MMP) and generation of reactive oxygen species (ROS) in Huh7.5 liver cancer cells. Proteomic and quantitative real-time PCR analyses indicated that EPS364 inhibited cancer cell growth and adhesion via targeting the FGF19-FGFR4 signaling pathway. These findings suggest that EPS364 is a promising antitumor agent for pharmacotherapy.