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。     

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.     

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.     

沙蒿多糖组合制剂对滩羊羔羊瘤胃菌群多样性的影响

本试验旨在研究沙蒿多糖不同组合制剂对滩羊羔羊瘤胃菌群多样性的影响。选择健康、体重相近的断奶滩羊公羔60只,按体重随机分为4个组,每组15只羊。对照组饲喂基础饲粮,试验1组、试验2组和试验3组分别饲喂在基础饲粮中添加5.5 g/d沙蒿多糖+5.5 g/d包被丁酸钠、5.5 g/d沙蒿多糖+5.5 g/d丁酸甘油酯和5.5 g/d沙蒿多糖+1.1 g/d莫能菌素的试验饲粮。预试期为15 d,正试期为60 d。结果表明:1)与对照组相比,试验1组和试验2组滩羊瘤胃液乙酸、丁酸、总挥发性脂肪酸浓度和乙酸比例显著提高(P<0.05)。2)试验2组的Chao1指数显著高于对照组(P<0.05),试验2组的Shannon指数和Simpson指数有高于对照组的趋势(0.05≤P<0.10)。3)门水平上,与对照组相比,试验1组瘤胃液中软壁菌门的相对丰度显著升高(P<0.05),厚壁菌门和丝状杆菌门的相对丰度有升高的趋势(0.05≤P<0.10);试验2组瘤胃液中绿弯菌门的相对丰度显著升高(P<0.05),而疣微菌门的相对丰度有升高的趋势(0.05≤P<0.10);试验3组瘤胃液中蓝藻菌门和互养菌门的相对丰度显著升高(P<0.05)。4)属水平上,与对照组相比,试验1组瘤胃液中理研菌科RC9的相对丰度显著降低(P<0.05),解琥珀酸菌属和丹毒丝菌属的相对丰度显著升高(P<0.05);试验2组瘤胃液中Sharpea的相对丰度显著降低(P<0.05),假丁酸弧菌属的相对丰度显著升高(P<0.05);试验3组瘤胃液中理研菌科RC9和Sharpea的相对丰度显著降低(P<0.05),互营球菌属的相对丰度有增加的趋势(0.05≤P<0.10)。由此可见,饲粮添加沙蒿多糖组合制剂均可提高滩羊羔羊菌群多样性,改善瘤胃微生物区系,且以添加沙蒿多糖+包被丁酸钠的效果最好。     

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.     

火龙果茎多糖的超高压提取及抗氧化活性研究

为研究火龙果茎多糖的超高压提取工艺及抗氧化活性。在单因素试验基础上,采用Box-Behnken设计和响应面分析方法,确定超高压提取的最优工艺,并从清除ABTS自由基和DPPH自由基能力方面来评价火龙果茎多糖的体外抗氧化能力。结果表明,火龙果茎多糖的超高压提取的最佳工艺条件为:液固比10∶1(m L∶g)、超高压压力300 MPa、超高压时间4 min。在此工艺条件下多糖得率为(2.83±0.02)%。火龙果茎对ABTS自由基和DPPH自由基具有清除作用,对ABTS自由基和DPPH自由基的清除率IC_(50)分别为浓度为4.3 mg/m L和5.5 mg/m L时。     

高抗氧化活性lager型啤酒酵母选育

采用过氧化氢刺激lager型啤酒酵母,Tadpoling法筛选细胞活力恢复较快突变菌,根据各菌株线粒体膜电位、胞内氧自由基(ROS)水平筛选获得4株活性较高菌株。比较突变菌与原始菌传代发酵性能、胞内ROS、活力指标及每一代细胞死亡率水平,最终获得一株抗氧化能力提高且活力稳定性较高啤酒酵母菌株。分析验证突变菌线粒体DNA修复相关基因MHR1发现,该基因发生部分碱基突变。