Sensitivity to Cisplatin in Head and Neck Cancer Cells Is Significantly Affected by Patient-Derived Cancer-Associated Fibroblasts

Cancer-associated fibroblasts (CAFs) are one of the most abundant and critical components of the tumor stroma. CAFs can impact many important steps of cancerogenesis and may also influence treatment resistance. Some of these effects need the direct contact of CAFs and cancer cells, while some involve paracrine signals. In this study, we investigated the ability of head and neck squamous cell carcinomas (HNSCC) patient-derived CAFs to promote or inhibit the colony-forming ability of HNSCC cells. The effect of cisplatin on this promoting or inhibiting influence was also studied. The subsequent analysis focused on changes in the expression of genes associated with cancer progression. We found that cisplatin response in model HNSCC cancer cells was modified by coculture with CAFs, was CAF-specific, and different patient-derived CAFs had a different “sensitizing ratio”. Increased expression of VEGFA, PGE2S, COX2, EGFR, and NANOG in cancer cells was characteristic for the increase of resistance. On the other hand, CCL2 expression was associated with sensitizing effect. Significantly higher amounts of cisplatin were found in CAFs derived from patients who subsequently experienced a recurrence. In conclusion, our results showed that CAFs could promote and/or inhibit colony-forming capability and cisplatin resistance in HNSCC cells via paracrine effects and subsequent changes in gene expression of cancer-associated genes in cancer cells.     

调控pH促进污泥产酸及两相耦合系统定向产乙酸

应用产氢产酸/同型产乙酸两相耦合工艺对市政污泥进行厌氧发酵,以实现高效产乙酸的目的.首先通过摇瓶试验考察了pH(7、8、9、10、11)对污泥产酸产气效能的影响,结果显示:pH=9为最佳产酸条件,乙酸最高浓度达8.23 g/L,产乙酸速率为0.56 g/(L·d);pH=7为最佳产气条件,产气速率为35.5 mL/(L·d).而后调节产氢产酸相(A相)的pH值使两相耦合工艺先后在最佳产酸及最佳产气条件下运行,对比乙酸总产量的差异.结果表明:系统在pH=9和pH=7下运行时所产乙酸总量分别为47.9、35.1 g,前者较后者高36.5%;虽然pH=7时能显著提高同型产乙酸相(H相)的产酸效能,但不足以抵消A相产酸不利带来的损失.可见,调控产氢产酸相的pH值为9可促进污泥产酸并实现污泥发酵定向产乙酸.     

混菌发酵产共轭亚油酸条件的优化

从淘米水中筛选出一株能高产共轭亚油酸（CLA）的菌株,通过菌株的菌落形态特征及微生物生理生化实验,确定其为植物乳杆菌（Lactobacillus plantarum）。以该植物乳杆菌和实验室保藏的嗜酸乳杆菌（lactobacillus acidophilus）为出发菌株,优化其混合发酵的培养条件。结果表明：底物添加量4.0%、菌种比例（植物乳杆菌：嗜酸乳杆菌）4：1、接种量5%、培养基初始pH5.5、34℃下发酵72h,CLA的合成量最高,可达到160.35μg/mL。     

Anaerobic CO2 fixation by the acetogenic bacterium Moorella thermoacetica

Anaerobic bacteria such as Moorella thermoacetica have the capacity of fixing carbon dioxide with carbon monoxide and hydrogen for the production of ethanol, acetic acid, and other useful chemicals. In this study, we evaluated the fixation of CO₂ for the production of acetic acid, as a product in its own right but also as precursor for lipid synthesis by oleaginous organisms. We achieved maximum cell optical density of 11.3, acetic acid titer of 31 g/L, and productivity of 0.55 g/L‐h at CO mass‐transfer rate of 83 mM/h. We also showed electron availability by CO mass transfer limited the process at CO mass transfer rates lower than 30 mM/h. Further enhancement of mass‐transfer rate removed such limitations in favor of biological kinetics as main limitation. This work underlines the potential of microbial processes for converting syngas to fuel and chemical products in processes suitable for distributed feedstock utilization. © 2013 American Institute of Chemical Engineers AIChE J, 59: 3176–3183, 2013     

大肠杆菌偏利共培养系统合成大豆苷元

大豆苷元是一种植物雌激素,具有多种生物活性,但在大肠杆菌中的生物全合成还未见报道。基于大豆苷元合成途径的三个模块（对香豆酸、甘草素和大豆苷元模块）,构建大肠杆菌共培养系统从头合成大豆苷元。将对香豆酸和甘草素模块分配到两株大肠杆菌中构建双菌共培养系统,合成甘草素。在此基础上,探索了三种共培养模式合成大豆苷元,结果显示,三菌共培养系统比其他两种双菌共培养系统的产量更高,达到27.8 mg/L。共培养菌株间通过苯丙氨酸的单向流动形成了偏利共生的关系,有助于平衡代谢途径,提高大豆苷元产量。该共培养系统在大肠杆菌中实现大豆苷元的从头合成,为其他黄酮类化合物的生物合成提供了即插即用的平台。     

一个产氢产乙酸菌互营共培养体的培养基优化

营养生态位位于产酸发酵菌和产甲烷菌之间的产氢产乙酸菌,其分离培养困难,种子资源匮乏,限制了基于强化产氢产乙酸功能作用的高效厌氧生物处理技术的开发．在前期获得对丙酸具有较强降解能力的产氢产乙酸茵互营共培养体7-m-2a的基础上,探讨了丙酸质量浓度、氮源、ca^2＋、Fe^2＋、Mg^2＋和泛酸等对其生长代谢的影响．结果表...     

3D Printing of Cell-Container-Like Scaffolds for Multicell Tissue Engineering

The development of an engineered non-contact multicellular coculture model that can mimic the in vivo cell microenvironment of human tissues remains challenging. In this study, we successfully fabricated a cell-container-like scaffold composed of β-tricalcium phosphate/hydroxyapatite (β-TCP/HA) bioceramic that contains four different pore structures, including triangles, squares, parallelograms, and rectangles, by means of three-dimensional (3D) printing technology. These scaffolds can be used to simultaneously culture four types of cells in a non-contact way. An engineered 3D coculture model composed of human bone-marrow-derived mesenchymal stem cells (HBMSCs), human umbilical vein endothelial cells (HUVECs), human umbilical vein smooth muscle cells (HUVSMCs), and human dermal fibroblasts (HDFs) with a spatially controlled distribution was constructed to investigate the individual or synergistic effects of these cells in osteogenesis and angiogenesis. The results showed that three or four kinds of cells cocultured in 3D cell containers exhibited a higher cell proliferation rate in comparison with that of a single cell type. Detailed studies into the cell–cell interactions between HBMSCs and HUVECs revealed that the 3D cell containers with four separate spatial structures enhanced the angiogenesis and osteogenesis of cells by amplifying the paracrine effect of the cocultured cells. Furthermore, the establishment of multicellular non-contact systems including three types of cells and four types of cells, respectively, cocultured in 3D cell containers demonstrated obvious advantages in enhancing osteogenic and angiogenic differentiation in comparison with monoculture modes and two-cell coculture modes. This study offers a new direction for developing a scaffold-based multicellular non-contact coculture system for tissue regeneration.     

Cre mRNA Is Not Transferred by EVs from Endothelial and Adipose-Derived Stromal/Stem Cells during Vascular Network Formation

Coculture systems employing adipose tissue-derived mesenchymal stromal/stem cells (ASC) and endothelial cells (EC) represent a widely used technique to model vascularization. Within this system, cell–cell communication is crucial for the achievement of functional vascular network formation. Extracellular vesicles (EVs) have recently emerged as key players in cell communication by transferring bioactive molecules between cells. In this study we aimed to address the role of EVs in ASC/EC cocultures by discriminating between cells, which have received functional EV cargo from cells that have not. Therefore, we employed the Cre-loxP system, which is based on donor cells expressing the Cre recombinase, whose mRNA was previously shown to be packaged into EVs and reporter cells containing a construct of floxed dsRed upstream of the eGFP coding sequence. The evaluation of Cre induced color switch in the reporter system via EVs indicated that there is no EV-mediated RNA transmission either between EC themselves or EC and ASC. However, since Cre mRNA was not found present in EVs, it remains unclear if Cre mRNA is generally not packaged into EVs or if EVs are not taken up by the utilized cell types. Our data indicate that this technique may not be applicable to evaluate EV-mediated cell-to-cell communication in an in vitro setting using EC and ASC. Further investigations will require a functional system showing efficient and specific loading of Cre mRNA or protein into EVs.     

Direct ethanol production from rice straw by coculture with two high-performing fungi

To develop efficient and economical direct ethanol production from fine rice straw crashed mechanically, two high-performing fungi, which can secret hyperactive cellulases and/or ferment effectively various sugars, were selected from some strains belong to Mucor circinelloides preserved in our laboratory. The simultaneous saccharification and fermentation (SSF) by coculture with these fungi was investigated. The screening of high-performing fungi resulted in the selection of NBRC 4572 as an ethanol-producing fungus and NBRC 5398 as a cellulase-secreting fungus. The strain 4572 produced ethanol aerobically from glucose and xylose in high yields of 0.420 g/g at 36 h and 0.478 g/g at 60 h, respectively, but secreted fairly low cellulases. On the other hand, the strain 5398 also produced ethanol from glucose in yield of 0.340 g/g though it had a little growth in xylose culture. However, it secreted hyperactive cellulases that are essential for hydrolysis of rice straw in culture and the maximum activities of endo-β-glucanase and β-glucosidase were 2.11 U/L and 1.47 U/L, respectively. In SSF of rice straw by coculture with two fungi selected, the ethanol production reached 1.28 g/L after 96 h when the inoculation ratio of the strain 5398 to the strain 4572 was 9.     

混菌固定化对生物合成共轭亚油酸的影响

海藻酸钠为固定材料,研究了嗜酸乳杆菌和嗜热链球菌先混菌后固定化对生物合成共轭亚油酸的影响.试验结果表明,最佳固定化条件为:海藻酸钠3.5%,氯化钙2.5%,菌种(嗜酸乳杆菌∶嗜热链球菌=1∶1,La:St=1∶1)包埋量40%,胶粒直径3mm,固定化时间1h;最佳发酵条件:培养温度37℃,发酵时间24h.在此条件下CLA积累量可达118.91μg/mL.固定化后菌种活力得到保持持久且可重复利用.