Qi Liu

Analysis of PM2.5-induced cytotoxicity in human HaCaT cells based on a microfluidic system

Human exposure to PM2.5 causes several adverse health effects. Skin is the first barrier against harmful environmental substances and can directly contact with PM2.5, but there is no study about PM2.5-induced cytotoxicity in human skin cells on the molecular level partially due to the shortcomings of traditional research methods. In present study, we established a microfluidic system including a cell culture chip integrated with a high-throughput protein microarray chip to investigate the mechanism of PM2.5-mediated cytotoxicity in human HaCaT cells. We found that PM2.5 was lodged inside the cytoplasm, mitochondria and nucleus of HaCaT cells by TEM. Flow cytometry analysis indicated that the cell apoptosis rate increased from 0.49% to 53.4%. The results of protein microarray showed that NF-κB and NALP3 signal transductions were activated in HaCaT cells after PM2.5 stimulations, up-regulating the expression of IL-1β and IL-6, which resulted in inflammatory response in HaCaT cells. Our findings provide a molecular insight into PM2.5-induced skin injury.

Rapid Capture and Analysis of Airborne Staphylococcus aureus in the Hospital Using a Microfluidic Chip

In this study we developed a microfluidic chip for the rapid capture, enrichment and detection of airborne Staphylococcus (S.) aureus. The whole analysis took about 4 h and 40 min from airborne sample collection to loop-mediated isothermal amplification (LAMP), with a detection limit down to about 27 cells. The process did not require DNA purification. The chip was validated using standard bacteria bioaerosol and was directly used for clinical airborne pathogen sampling in hospital settings. This is the first report on the capture and analysis of airborne S. aureus using a novel microfluidic technique, a process that could have a very promising platform for hospital airborne infection prevention (HAIP).

An in vitro cytotoxicities comparison of 16 priority polycyclic aromatic hydrocarbons in human pulmonary alveolar epithelial cells HPAEpiC

In present study, we compared for the first time the cytotoxicities of the 16 priority polycyclic aromatic hydrocarbons (PAHs) in human pulmonary alveolar epithelial cells HPAEpiC. Moreover, we examined the effects of each PAH on oxidative stress (SOD, GSH, and ROS), cell viability, extracellular LDH, and apoptosis. The 16 priority PAHs were classified into four levels of cytotoxicity: (1) high cytotoxicity, BkF, BaP, and DBA; (2) moderate cytotoxicity, BbF, IND, BghiP, BaA, and CHR; (3) low cytotoxicity, PA, FL, and Pyr; and (4) mild cytotoxicity, Nap, AcPy, Acp, Flu, and Ant. Most of the PAHs showed benzene-ring-related cytotoxicity, except PA with 3-ring structure, cytotoxicity of which is similar to those of FL and Pyr with 4-ring structure. Results indicated the need for more studies on DBA, IND, and BghiP, among others, which are rarely investigated. PA, FL, and Pyr with little carcinogenicity should also be evaluated. This study will provide useful references for studies on the effects of PAHs on different cells or animal models.

Signal Transductions of BEAS-2B Cells in Response to Carcinogenic PM2.5 Exposure Based on a Microfluidic System

PM2.5 (particulate matter less than 2.5 μm in diameter) is considered as a harmful carcinogen. Determining the precise relationship between the chemical constituents of PM2.5 in the air and cancer progression could aid the treatment of environment related disease and establishing risk reduction strategies. Herein, we used transcriptomics (RNA-seq) and an integrated microfluidic system to identify the global gene expression and differential target proteins expression induced by ambient fine particles collected from the heavy haze in China. The results clearly indicated that cancer related pathways exhibited the strongest dysregulation. The ambient fine particles could be uptaken into the cells by pinocytosis, mainly promoting the PI3K-Akt pathway, FGF/FGFR/MAPK/VEGF signaling, and the JAK-STAT pathway, leading to evading apoptosis, sustained angiogenesis, and cell proliferation, which are the most important hallmarks of cancer. And fine particles also have been demonstrated to create intracellular reactive oxygen species (ROS) and mitochondrial ROS, change intracellular free Ca2+, and induce apoptosis, which are all key players in mediating cancer progression. It was observed by transmission electron microscopy (TEM) that the particles from the haze could enter the mitochondria, resulting in disturbance of the mitochondrial membrane and disruption of the mitochondria, and these particles can even enter inside the nucleus. It was also found in our study of organics (OC, PAHs) and metals (Zn, As, V) that compounds of fine particles were more closely associated with the exacerbation of cancer and secondary aerosols generated by traffic had the largest impact on cancer related signal transductions.

Expression of Acidothermus cellulolyticus thermostable cellulases in tobacco and rice plants

ABSTRACT The production of cellulases in plants is an economical method for the conversion of lignocellulosic biomass into fuels. Herein we report the expressions of two thermostable Acidothermus cellulolyticus cellulases, endo-1,4-β-D-glucanase (E1) and exoglucanase (Gux1), in tobacco and rice. To evaluate the expression of these recombinant cellulases, we expressed the full-length E1, the catalytic domains of E1 (E1cd) and Gux1 (Gux1cd), as well as an E1–Gux1cd fusion enzyme in various subcellular compartments. In the case of tobacco, transgenic plants that expressed apoplast-localized E1 showed the highest level of activity, about three times higher than those that expressed the cytosolic E1. In the case of rice, the level of cellulase-specific activity in the transgenic plants ranged from 11 to 20 nmol 4-methylumbelliferone min−1 mg−1 total soluble protein. The recombinant cellulases exhibited good thermostability below 70 °C. Furthermore, transgenic rice leaves that were stored at room temperature for a month lost about 20% of the initial cellulase activity. Taken together, the results suggested that heterologous expression of thermostable cellulases in plants may be a viable option for biomass conversion.

A microfluidic chip for studying the reproduction of Enteromorpha prolifera

In recent years, green tides caused by water eutrophication, has brought serious environmental problems. Enteromorpha prolifera (E. prolifera), an opportunistic macroalgae, is one of the main source contributing to the formation of green tides. It has been estimated that the excessive growth of E. prolifera is closely related to various reproductive ways of germ cells which are at the micrometer scale. Here we report a microfluidic device named Germ Cell Capture Chip (GCChip) to investigate the E. prolifera reproductive mechanism. GCChip integrates the functions of algal growing, and the release, capture and selective culture of germ cells. Automatic separation and capture of germ cells on the chip allows to study germ cells response to different stimuli. The novel device greatly facilitates long-term live-cell imaging at cellular resolution and implements the rapid and accurate exchange of growth medium without manual intervention. Results showed that the starting time of germ cell releases were earlier on the chip than that of traditional experiments with more concentrated breakout. Moreover, GCChip can be widely applied on the study of other algae. The study of algae growth process, including the elongation of somatic cell, the generation, and the release of reproductive cells, can all be improved by using this microfluidic platform.

Cytotoxicity analysis of indoor air pollution from biomass combustion in human keratinocytes on a multilayered dynamic cell culture platform.

Skin tissue is the first barrier against ambient harmful matter and has direct contact with indoor air pollutants. Nevertheless, a comprehensive understanding of cytotoxicity of indoor air pollution on skin cells is insufficiently clear. Herein, for the first time a multilayered dynamic cell culture platform was established to study the cytotoxicity of indoor air pollutant from biomass combustion in human skin keratinocytes. The platform consisted of seven repetitive polydimethylsiloxane modules carrying six pieces of polycarbonate membrane between them as substrate for cell growth to realize the simultaneous dynamic culture of 12 layers of keratinocytes. After exposure to biomass combustion soluble constituents (BCSCs), cell viability under microfluidic platform conditions declined more significantly, and apoptosis rates increased more obviously compared with well plate conditions. Transmission electron microscope showed that keratinocyte microstructures displayed obvious signs of cellular damage. Our study confirmed that the nuclear factor of kappa B (NF-κB) signaling pathway was activated, which significantly increased the Bax/Bcl-2 ratio and tumor necrosis factor-alpha and interleukin 6 expression, indicating that NF-κB signaling pathway was the major factor in BCSCs-induced cytotoxicity. These findings offer an insight into the mechanism of BCSCs-induced cytotoxicity in keratinocytes and provide a theoretical basis for future studies on skin cells.

A microfluidic system for rapid detection of airborne pathogenic fungal spores

Airborne fungi, including Aspergillus species, are the major causes of human asthma. Direct capture and analysis of pathogenic fungi in indoor air is important for disease prevention and control. In this paper, we demonstrated an integrated microfluidic system capable of enrichment and high-throughput detection for airborne fungal spores of Aspergillus niger, a well-known allergenic harmful species. The microfluidic system allowed semiquantitative detection of Aspergillus niger spores based on immunofluorescence analysis. To assess its contaminated level, the whole analysis time could be completed in 2-3 h including ∼1 h of enrichment and ∼1 h of target detection. The detection limit was ∼20 spores, equivalent to ∼300 spores·m-3 of the concerned targets in air. In addition, the microfluidic system has integrated sampling and sample analysis to avoid additional sample concentration step, showing the potential for point-of-care detection for other pathogenic fungal spores.