Expression of Anti-Lipopolysaccharide Factor Isoform 3 in Chlamydomonas reinhardtii Showing High Antimicrobial Activity

Antimicrobial peptides are a class of proteins with antibacterial functions. In this study, the anti-lipopolysaccharide factor isoform 3 gene (ALFPm3), encoding an antimicrobial peptide from Penaeus monodon with a super activity was expressed in Chlamydomonas reinhardtii, which would develop a microalga strain that can be used for the antimicrobial peptide production. To construct the expression cluster, namely pH2A-Pm3, the codon optimized ALFPm3 gene was fused with the ble reporter by 2A peptide and inserted into pH124 vector. The glass-bead method was performed to transform pH2A-Pm3 into C. reinhardtii CC-849. In addition to 8 μg/mL zeocin resistance selection, the C. reinhardtii transformants were further confirmed by genomic PCR and RT-PCR. Western blot analysis showed that the C. reinhardtii-derived ALFPm3 (cALFPm3) was successfully expressed in C. reinhardtii transformants and accounted for 0.35% of the total soluble protein (TSP). Furthermore, the results of antibacterial assay revealed that the cALFPm3 could significantly inhibit the growth of a variety of bacteria, including both Gram-negative bacteria and Gram-positive bacteria at a concentration of 0.77 μM. Especially, the inhibition could last longer than 24 h, which performed better than ampicillin. Hence, this study successfully developed a transgenic C. reinhardtii strain, which can produce the active ALFPm3 driven from P. monodon, providing a potential strategy to use C. reinhardtii as the cell factory to produce antimicrobial peptides.     

杏鲍菇秋葵咀嚼片配方优化与质量标准研究

为了优选杏鲍菇秋葵咀嚼片配方,以杏鲍菇粗多糖、杏鲍菇超细粉为主要原料,采用全粉末直接压片技术,选取片重差异、崩解时限、硬度及脆碎度的综合评定值为评价指标,通过响应面法确定最佳配方,并制定产品的质量指标。结果表明,杏鲍菇秋葵咀嚼片的最佳配方为杏鲍菇粗多糖38%、杏鲍菇超细粉49%和黄秋葵超细粉13%,制成的咀嚼片表面光滑、色泽均匀。综上,采用多指标综合评分法优化杏鲍菇秋葵咀嚼片配方是可行的,所得工艺配方稳定可靠,这为我国咀嚼片剂工艺创新及其产业化应用研究提供了新思路。     

菠萝蜜种子淀粉制备香草兰微胶囊的工艺研究

以菠萝蜜种子淀粉为壁材,采用饱和水溶液法制备香草兰精油微胶囊,以包埋产率为指标,采用响应面分析法对微胶囊的包埋条件进行优化探讨。结果表明:5个单因素中,影响最显著的因素为壁芯材比例、包埋温度和包埋时间。响应面优化得到香草兰精油微胶囊的最佳工艺条件为壁芯材比例为7.5∶1;包埋时间72 min;包埋温度54℃,此条件下的包埋产率为(95.46±0.2)%,包埋率为(76.35±0.6)%,载油量为(27.73±0.3)%。试验证明,此条件结果与模型预测值相吻合,此工艺条件可为菠萝蜜种子淀粉包埋香草兰精油微胶囊工艺提供理论依据。     

甘蔗叶不同还田方式对土壤养分的影响

为减轻秸秆焚烧所带来的环境问题和充分利用甘蔗叶资源,在大田环境下动态监测甘蔗叶粉碎、焚烧和深埋3种还田方式对土壤养分的影响。结果表明:甘蔗叶粉碎还田和深埋还田有利于甘蔗叶中有机碳及氮、磷、钾养分的缓慢释放;甘蔗叶焚烧还田提高了土壤碱解氮含量5.6~20.9 mg/kg和速效磷含量0.8~4.0 mg/kg,促进土壤中氮磷向有效态转化,焚烧还田初期显著增加了土壤速效钾的含量12.3~18.3 mg/kg,但焚烧会损失部分甘蔗叶中碳氮元素,也造成环境污染。综合分析认为甘蔗叶粉碎还田是比较省工、方便的处理方式。     

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.     

不同处理方式对蒸谷米复水性的影响

以稻谷为原料,对蒸谷米的复水性进行了研究。通过单因素试验,探讨超高压浸泡工序、蒸煮工序、干燥工序中相关参数的影响规律,并以蒸煮时柠檬酸添加量、蒸煮次数、微波干燥时间为变量,成品复水率为指标,通过响应面分析建立回归方程,得出蒸煮次数对复水性影响最大,柠檬酸添加量次之,干燥时间影响最小。利用模型确定了较佳参数为柠檬酸添加量1.1%,微波干燥时间6.4 min,蒸煮次数3次。     

响应面法优化超声波辅助提取薏苡仁低聚糖工艺的研究

采用响应面法优化超声波辅助提取薏苡仁低聚糖的工艺条件。在单因素试验基础上,选取液料比、超声波时间以及超声波功率3个因素结合Box-Behnken试验建立数学模型,分析考察3个因素对薏苡仁低聚糖响应值的影响程度,优化工艺参数。各因素对薏苡仁低聚糖提取率影响程度从大到小顺序依次为:超声波功率超声波时间液料比。响应面设计法优化出其最佳超声波提取条件为:超声波温度70℃,液料比33∶1(m L/g),超声波时间27 min,超声波功率450 W。在该条件下,薏苡仁低聚糖提取率为0.94%,与模型预测值0.98%接近。说明使用响应面法优化超声波辅助提取薏苡仁低聚糖的工艺条件是可行的。     

耕作方式及秸秆还田对土壤性质、微生物碳源代谢及小麦产量的影响

以小麦品种济麦22为试验材料,采用裂区试验设计,耕作方式为主区,分别设常规翻耕(C)、深松(S)、旋耕(R)处理,副区为秸秆还田量,分别设秸秆全还田(P)和秸秆不还田(A)处理,采用Biolog Eco技术测定土壤微生物碳源代谢功能,并分析土壤基本理化性质和作物产量。结果显示:深松与秸秆还田均有利于土壤含水量和有机碳含量的提高,0～15 cm土层分别提高了9.78%和24.00%,15～30 cm土层分别提高了7.08%和15.81%;深松提高了15～30 cm土层的pH值6.67%,秸秆还田提高了0～15 cm土层的pH值4.32%。深松和秸秆还田均有利于代谢多样性(丰富度指数、香浓多样性指数)、碳源代谢强度的提高,0～15 cm土层分别提高了26.84%、3.84%和38.02%,15～30 cm土层分别提高了11.87%、 3.63%和14.74%。主成分分析表明常规翻耕秸秆不还田和旋耕耕作秸秆不还田碳源代谢功能相近,15～30 cm层次内常规翻耕秸秆全还田碳源代谢功能和深松耕作秸秆全还田处理相近。深松和秸秆还田平均提高了小麦产量5.82%,微生物碳源代谢功能与小麦产量具有极显著的相关性。