Bakuchiol inhibits lung cancer by modulating tumor microenvironment and the expression of PD-L1

Immune checkpoint therapy is an emerging frontier in cancer therapy. With the aim to develop an efficient herb derived compound to facilitate immune checkpoint therapy, here we investigate if a herb-derived compound, Bakuchiol (BAK), can be used to treat lung cancer and elucidate if BAK could serve as a PD-L1 regulator. To this end, a murine lung cancer model was established by subcutaneously inoculating murine Lewis lung carcinoma (LLC) cells. BAK of 5 to 40 mg/kg was used for treatment in vivo for 15 days. On Day 15, the population of CD4+ and CD8+ T cells, Treg cells. BAK could effectively inhibit tumor growth by starting treatment either on Day 0 or 6 after tumor inoculation at doses of 5−40 mg/kg. BAK treatment increased the population of cytotoxic immune cells (i.e., CD8+ T cells, and M1 macrophages), meanwhile decreasing pro-tumor immune cells (i.e., CD3+ T cells, Treg cells, and M2 macrophages). Anti-inflammatory cytokines, including IL1β, IL2, IFNγ, TNF-α, IL4 and IL10 were upregulated by BAK. PD-L1 expression in the tumor was also lowered by BAK. AKT and STAT3 signaling were inhibited by BAK. BAK is an efficient agent in reducing LLC tumor growth. These data support the potential of BAK as a new drug for treating lung cancer by serving as a PD-L1 inhibitor that suppresses the activation of AKT and STAT3.     

URA3基因影响青蒿酸酿酒酵母工程菌中试发酵产量

CRISPR/Cas9基因编辑技术已经被广泛应用于工程酿酒酵母的基因插入、基因替换和基因敲除,通过使用选择标记进行基因编辑具有简单高效的特点。前期利用CRISPR/Cas9系统敲除青蒿酸生产菌株酿酒酵母(Saccharomyces cerevisiae) 1211半乳糖代谢负调控基因GAL80,获得菌株S. cerevisiae 1211-2,在不添加半乳糖诱导的情况下,青蒿酸摇瓶发酵产量达到了740 mg/L。但在50 L中试发酵实验中,S. cerevisiae 1211-2很难利用对青蒿酸积累起到决定性作用的碳源-乙醇,青蒿酸的产量仅为亲本菌株S.cerevisiae 1211的20%–25%。我们推测因遗传操作所需的筛选标记URA3突变,影响了其生长及青蒿酸产量。随后我们使用重组质粒pML104-KanMx4-u连同90 bp供体DNA成功恢复了URA3基因,获得了工程菌株S. cerevisiae 1211-3。S. cerevisiae 1211-3能够在葡萄糖和乙醇分批补料的发酵罐中正常生长,其青蒿酸产量超过20g/L,与亲本菌株产量相当。研究不但获得了不加半乳糖诱导的青...     

迷迭香挥发物不同组合对假眼小绿叶蝉行为的调控

为探讨茶园群落中非茶植物气味引诱或驱避重要茶树害虫假眼小绿叶蝉的效应,遂将茶园常见杂草迷迭香挥发物中樟脑、α-松油醇和石竹烯分别配成10-2g/m L剂量,再等量组成混合物,在每个橡皮头上载100#L制成诱芯,附于素馨黄粘板上,于茶园中诱捕假眼小绿叶蝉,发现这种三组分诱芯明显地驱避假眼小绿叶蝉。将行为反应中分别对假眼小绿叶蝉呈现显著引诱效应的10-2g/m L樟脑、10-4g/m L石竹烯、10-6g/m Lα-水芹烯、10-4g/m Lα-松油醇和10-8g/m L桉树脑等量配成五组分混合物,同法制成诱芯,附于素馨黄粘板,诱捕假眼小绿叶蝉。发现五组分诱芯:(1)显著引诱假眼小绿叶蝉,诱效稍强于现价段茶园中使用的源于茶梢挥发物的诱芯;(2)每日诱捕的假眼小绿叶蝉数与天数呈抛物线关系;(3)加入液体石蜡作为缓释剂,可将诱效的半衰期延长2.5—4.0d;(4)18:00—6:00诱得的假眼小绿叶蝉数量占全天的70%—75%。分析认为:非茶植物迷迭香的挥发物中含有显著引诱假眼小绿叶蝉信息物质,这类信息物质制成的五组分诱芯的诱效较强,可以作为一种防治假眼小绿叶蝉的手段,早晨和黄昏是最佳诱捕时分。     

2,5-Hexanedione induced apoptosis in rat spinal cord neurons and VSC4.1 cells via the proNGF/p75NTR and JNK pathways

Increasing evidence suggests that n-hexane induces nerve injury via neuronal apoptosis induced by its active metabolite 2,5-hexanedione (HD). However, the underlying mechanism remains unknown. Studies have confirmed that pro-nerve growth factor (proNGF), a precursor of mature nerve growth factor (mNGF), might activate apoptotic signaling by binding to p75 neurotrophin receptor (p75NTR) in neurons. Therefore, we studied the mechanism of the proNGF/p75NTR pathway in HD-induced neuronal apoptosis. Sprague–Dawley (SD) rats were injected with 400 mg/kg HD once a day for 5 weeks, and VSC4.1 cells were treated with 10, 20, and 40 mM HD in vitro. Results showed that HD effectively induced neuronal apoptosis. Moreover, it up-regulated proNGF and p75NTR levels, activated c-Jun N-terminal kinase (JNK) and c-Jun, and disrupted the balance between B-cell lymphoma-2 (Bcl-2) and Bcl-2-associated X protein (Bax). Our findings revealed that the proNGF/p75NTR signaling pathway was involved in HD-induced neuronal apoptosis; it can serve as a theoretical basis for further exploration of the neurotoxic mechanisms of HD.     

Production of uridine 5′-monophosphate by Corynebacterium ammoniagenes ATCC 6872 using a statistically improved biocatalytic process

Attempts were made with success to develop a two-step biocatalytic process for uridine 5′-monophosphate (UMP) production from orotic acid by Corynebacterium ammoniagenes ATCC 6872: the strain was first cultivated in a high salt mineral medium, and then cells were harvested and used as the catalyst in the UMP production reaction. Effects of cultivation and reaction conditions on UMP production were investigated. The cells exhibited the highest biocatalytic ability when cultivated in a medium containing corn steep liquor at pH 7.0 for 15 h in the exponential phase of growth. To optimize the reaction, both “one-factor-at-a-time” method and statistical method were performed. By “one-factor-at-a-time” optimization, orotic acid, glucose, phosphate ion (equimolar KH₂PO₄ and K₂HPO₄), MgCl₂, Triton X-100 were shown to be the optimum components for the biocatalytic reaction. Phosphate ion and C. ammoniagenes cell were furthermore demonstrated as the most important main effects on UMP production by Plackett–Burman design, indicating that 5-phosphoribosyl-1-pyrophosphate (PRPP) synthesis was the rate-limiting step for pyrimidine nucleotides production. Optimization by a central composition design (CCD) was then performed, and up to 32 mM (10.4 g l⁻¹) UMP was accumulated in 24 h from 38.5 mM (6 g l⁻¹) orotic acid. The yield was threefold higher than the original UMP yield before optimization.     

卡西霉素生物合成基因簇中调控基因calR1的功能

【背景】卡西霉素(calcimycin)是重要的离子载体抗生素,其生物合成基因簇已从教酒链霉菌NRRL3882的基因组DNA中成功克隆,但基因簇内的部分生物合成基因及调控基因的功能有待研究。【目的】研究卡西霉素产生菌教酒链霉菌NRRL3882中编码TylR家族同源转录调控蛋白的calR1基因的功能。【方法】通过PCR-targeting的方法,构建calR1基因敲除突变株及回补菌株,对突变菌株及回补菌株进行发酵,通过HPLC分析其代谢产物。利用荧光定量PCR检测ΔcalR1突变菌株和野生菌株的生物合成基因转录水平。【结果】calR1基因敲除突变株丧失产生卡西霉素的能力,但仍有中间产物噻唑霉素的积累,回补菌株中卡西霉素的产量有一定程度的恢复。RT-qPCR结果表明,卡西霉素合成相关的一些重要基因calC、calG、calU3等基因的表达量明显改变。【结论】TylR家族转录调控基因calR1是卡西霉素生物合成的调控基因。     

秋末苏南茶园昆虫的群落组成及其趋色性

苏南地区名茶荟萃,而虫害历来较为严重。秋末选丹阳市一片无公害茶园,使用纯白、桃红、墨绿、果绿、湖蓝、天蓝、素馨黄、芽绿、土黄、桔黄、大红和紫色12种粘性色板诱虫。结果表明:① 4日内捕获7目42科85种30455头昆虫,其中优势类群是同翅目、膜翅目和双翅目,三者个体数分别占总个体数的86.5%、8.8%和2.5%。②主要害虫是假眼小绿叶蝉和茶蚜,分别占总个体数的15%和71.5%。③ 捕获的中华蜜蜂占总个体数8.2%。④捕获天敌昆虫1034头,其中,瓢虫类32头,占3.1%,主要种类是异色瓢虫和黄斑盘瓢虫;草蛉类582头,占56.3%,包括中华草蛉、大草蛉和丽草蛉;伞裙追寄蝇和蚕饰腹寄蝇等5种寄生蝇类111头,占10.7%;门氏食蚜蝇和黑带食蚜蝇等7种食蚜蝇类110头,占10.6%;螟蛉瘤姬蜂和花胸姬蜂等5种姬蜂类、茶尺蠖绒茧蜂和单白绵绒茧蜂等7种茧蜂类个体数分别占3.9%和9.4%。⑤ 芽绿、素馨黄和桔黄色板显著地引诱假眼小绿叶蝉和茶蚜;芽绿、素馨黄色板显著地引诱姬蜂和茧蜂;芽绿、土黄和果绿色板明显地引诱草蛉类;果绿、天蓝和紫色色板引诱较多的蝇类和蚊类等双翅目昆虫;素馨黄引诱的各类昆虫种数最多;纯白板上各类昆虫的多样性指数最大,表明其对许多昆虫都有引诱效应。秋末时节查明即将越冬的害虫和天敌昆虫种类、数量和益害比、以及优势种害虫和优势种天敌数量,探明多种色彩引诱益、害虫的差异,对于有效实施无公害封园防治,以压低越冬基数有重要意义。     

Microbial mediation of biogeochemical cycles revealed by simulation of global changes with soil transplant and cropping

Despite microbes'' key roles in driving biogeochemical cycles, the mechanism of microbe-mediated feedbacks to global changes remains elusive. Recently, soil transplant has been successfully established as a proxy to simulate climate changes, as the current trend of global warming coherently causes range shifts toward higher latitudes. Four years after southward soil transplant over large transects in China, we found that microbial functional diversity was increased, in addition to concurrent changes in microbial biomass, soil nutrient content and functional processes involved in the nitrogen cycle. However, soil transplant effects could be overridden by maize cropping, which was attributed to a negative interaction. Strikingly, abundances of nitrogen and carbon cycle genes were increased by these field experiments simulating global change, coinciding with higher soil nitrification potential and carbon dioxide (CO₂) efflux. Further investigation revealed strong correlations between carbon cycle genes and CO₂ efflux in bare soil but not cropped soil, and between nitrogen cycle genes and nitrification. These findings suggest that changes of soil carbon and nitrogen cycles by soil transplant and cropping were predictable by measuring microbial functional potentials, contributing to a better mechanistic understanding of these soil functional processes and suggesting a potential to incorporate microbial communities in greenhouse gas emission modeling.