Quantitative Analysis of Nanoplastics in Single Cells by Subcellular Chromatography.

The accumulation and spatial distribution of intracellular nanoplastic particles provide useful information about their spatiotemporal toxicological effects mediated by the physicochemical parameters of nanoplastics in living cells. In this study, a sample injection-transfer method was designed with an accuracy of up to femtoliters to attoliters to match the volume required for ultranarrow-bore open-tubular liquid chromatography. The separation and concentration quantification of mixed polystyrenes in different regions in living cells were achieved by directly transferring picoliter/femtoliter volumes of intracellular cytoplasm to an ultranarrow-bore open-tubular chromatographic column. The measurement of pollutant concentration in different areas of a small-volume target (single cell) was realized. This method is expected to be used in the qualitative and quantitative analyses of complex, mixed, and label-free nanoplastics (a few nm in size) in the subregions of living cells.

Exogenous GA3 application altered morphology, anatomic and transcriptional regulatory networks of hormones in Eucalyptus grandis.

Gibberellins (GAs) play a key role in plant growth and development including cell elongation, cell expansion, and xylem differentiation. Eucalyptus are the world's most widely planted hardwood trees providing fiber and energy. However, the roles of GAs in Eucalyptus remain unclear and their effects on xylem development remain to be determined. In this study, E. grandis plants were treated with 0.10 mg L-1 GA3 and/or paclobutrazol (PAC, a GA inhibitor). The growth of shoot and root were recorded, transverse sections of roots and stems were stained using toluidine blue, and expression levels of genes related to hormone response and secondary cell wall biosynthesis were analyzed by quantitative real-time PCR. The results showed that GA3 dramatically promoted the length of shoot and root, but decreased the diameter of root and stem. Exogenous GA3 application also significantly promoted xylem development in both stem and root. Expression analysis revealed that exogenous GA3 application altered the transcript levels of genes related to the GA biosynthetic pathway and GA signaling, as well as genes related to auxin, cytokinin, and secondary cell wall. These findings suggest that GAs may interact with other hormones (such as auxin and cytokinin) to regulate the expression of secondary cell wall biosynthesis genes and trigger xylogenesis in Eucalyptus plants.

Magnetic bead based immunoassay for enumeration of CD4+ T lymphocytes on a microfluidic device.

Human immunodeficiency virus (HIV) diagnostics are urgently needed in resource-scarce settings. Monitoring of HIV-infected patients requires accurate counting of CD4(+) T lymphocytes. However, the current methods for enumeration of CD4(+) T lymphocytes are of high cost, technically complex and time-consuming. In this paper, we developed a simple, rapid and inexpensive one-step immunomagnetic method for separating and counting CD4(+) T lymphocytes on microfluidic devices with enlarged reaction chambers. CD4(+) T lymphocytes were successfully separated and captured from the cell suspension obtained from mouse thymus. CD4 counts were determined under an optical microscope in a rapid and simple format. In order to acquire the maximum efficiency of cell capture, relative parameters were investigated, including section area of the reaction chamber and injection flow rate of the cell suspension. The enlarged reaction chamber with two symmetrical cone-shaped ends was helpful for cell capture, and the maximum capability of captured CD4(+) T lymphocytes was about 700 cells microL(-1). Our investigations avoided the complex sample pre-treatment, and the entire analysis time was significantly reduced to 15 min. This CD4 counting microdevice had the potential to reduce the cost for HIV diagnosis in resource-limited settings.

Microfluidic cell culture and metabolism detection with electrospray ionization quadrupole time-of-flight mass spectrometer.

A novel method for the characterization of drug metabolites was developed by integrating chip-based solid-phase extraction (SPE) with an online electrospray ionization quadrupole time-of-fight mass spectrometer (ESI-Q-TOF-MS). The integrated microfluidic device was composed of circular chambers for cell culture and straight microchannels with shrink ends to pack the solid-phase material for sample cleanup and concentration prior to mass analysis. By connecting the two separated microchannels with polyethylene tubes, drug metabolism studies related to functional units, including cell culture, metabolism generation, sample pretreatment, and detection, were all integrated into the microfluidic device. To verify the feasibility of a drug metabolism study on the microfluidic device, the metabolism of vitamin E in human lung epithelial A549 cells was studied. The metabolites were successfully detected by online ESI-Q-TOF-MS with high sensitivity and short analysis time (8 min). By integrating several parallel channels, the desalting and concentration process could be simultaneously achieved. The total sample pretreatment time only needed about 15 min, and solvent consumption could be reduced to less than 100 microL. All this demonstrated that the developed microfluidic device could be a potential useful tool for cellular drug metabolism research.

A microfluidic approach for anticancer drug analysis based on hydrogel encapsulated tumor cells.

A novel method based on fluorescence detection of hydrogel encapsulated cells in microchannels was developed for anticancer drug analysis. In this work, human hepatoma HepG2 cells and human lung epithelial A549 cells were simultaneously immobilized inside two different shapes of three-dimensional hydrogel microstructures using photolithography approach on a same array. Microarrays of living cells offer the potential for parallel detection of many cells and thereby enable high-throughput assays. Using a photolithographic setup, we investigated the prepolymer composition and crosslinking parameters that influenced cell viability inside photocrosslinked hydrogels. The viability of cells encapsulated inside hydrogel microstructures was higher than 90% under optimized photocrosslinking conditions. The cells were further cultured under stable conditions and remained viable for at least three days that were able to carry out cell-based assays. Furthermore, we studied the variation of two intracellular redox parameters (glutathione and reactive oxygen species) in anticancer drug-induced apoptosis in HepG2 and A549 cells. Two anticancer drugs exhibited distinct effects on the levels of intracellular glutathione and reactive oxygen species, indicating the selectivity of these drugs on the disturbance of redox balance within cells. The established platform provides a convenient and fast method for monitoring the effect of anticancer drugs on tumor cells, which is very useful for fundamental biomedical research.

[Study on X-ray powder diffraction of various structured zinc titanate prepared by the method of direct precipitation].

Zinc titanate powders were prepared from Ti(SO4)2, Zn(NO3)2 x (6)H2O and (NH4)2CO3 by the method of direct precipitation. The effects of reaction conditions on the structure of zinc titanate were studied. The sample was analyzed by means of XRD and TG-DTA. The structure of zinc titanate was affected by the reaction subsequence of the formation of titanic acid and zinc carbonate. In the reaction system where titanic acid was generated earlier, collision reaction occurred between the generated zinc carbonate molecule and the surrounding titanic acid molecule. When titanic acid was generated earlier and precipitant (NH4)2CO3 was sufficient, Zn2Ti3O8 was obtained because of the sufficient collision reaction and superfluous titanic acid. In the reaction system where zinc carbonate was generated earlier, collision reaction occurred between the generated titanic acid molecule and the surrounding zinc carbonate molecule. When zinc carbonate was generated earlier and precipitant (NH4)2CO3 was sufficient, Zn2TiO4 was obtained because of the sufficient collision reaction and superfluous zinc carbonate. In addition, the kinds and structure of the production were affected by the dosage of precipitant and the reaction temperature. Zn2Ti3O8 or Zn2TiO4 could be obtained easier when using more precipitant or higher reaction temperature which could cause more sufficient collision reaction. ZnTiO3 could be obtained under the conditions of less precipitant and lower reaction temperature.

Synthesis of titania and titanate nanomaterials and their application in environmental analytical chemistry.

TiO(2) nanoparticles and H(2)Ti(2)O(5).H(2)O, Na(2)Ti(2)O(4)(OH)(2) nanotubes were synthesized by solvothermal method and their applications in the degradation of active Brilliant-blue (KN-R) solution were investigated. The experimental results revealed that the synthesized TiO(2) nanoparticles had a good crystallinity and a narrow size distribution (about 4-5 nm); the obtained H(2)Ti(2)O(5).H(2)O, Na(2)Ti(2)O(4)(OH)(2) were tubelike products with an average diameter of approximately 20-30 and approximately 200-300 nm length. The three catalysts we synthesized had some hydroxyl groups and the maximum absorption boundaries of the samples were all red-shifted, which indicated the samples had a promising prospect in photocatalysis. The results of the photocatalytic experiments indicated that the photocatalytic activity of the samples was: TiO(2)>H(2)Ti(2)O(5).H(2)O>Na(2)Ti(2)O(4)(OH)(2), which was in good accordance with the fact of FTIR and UV-vis absorption spectra. The formation mechanism of these nanostructures was also discussed.

A new layered zinc phosphate templated by protonated isonicotinate, [Zn2(C6H5NO2)2(HPO4)2].

An organic-inorganic hybrid compound, poly[bis[(pyridine-4-carboxylato)zinc(II)]-di-mu3-phosphato], [Zn2(C6H5NO2)2(HPO4)2], has been hydrothermally synthesized and structurally characterized. The crystal structure consists of two types of two-dimensional layers of zinc hydrogenphosphate templated by protonated isonicotinate (ina) (or 4-pyridinecarboxylic acid), which contain two crystallographically independent centrosymmetric [Zn2(ina)2(HPO4)2] dimers as basic building units. The layers are interconnected via hydrogen-bonding and heterocyclic ring interactions.