Microfluidic Engineering of Addressable Multicompartmental Microspheres for Multicellular Systems.

With the advantages of high-throughput manufacturing and customizability, on-microsphere construction of in vitro multicellular analytical systems has garnered significant attention. However, achieving a precise, biocompatible cell arrangement and spatial signal analysis in hydrogel microspheres remains challenging. In this work, a microfluidic method is reported for the biocompatible generation of addressable supersegmented multicompartmental microspheres. Additionally, these microspheres are developed as novel label-free multicellular systems. In the microfluidic approach, controllable microfluidics is used to finely tune the internal microstructure of the microspheres, and the gas ejector ensures the biocompatibility of the preparation process. As a proof of concept, six- and twenty-compartment microspheres were obtained without the addition of any biohazardous reagents. For microsphere decoding, the visualization of two basic compartments can provide clues for identifying label-free cells due to the structural regularity of the microspheres. Finally, by encapsulating cells of different types, these microspheres as multicellular systems were successfully used for cell coculture and drug testing. These biocompatible, scalable, and analyzable microspheres will open up new prospects for biomedical analysis.

Fabrication and Performance of Bubble-Containing Multicompartmental Particles: Novel Self-Orienting Carriers.

In this work, a class of bubble-containing multicompartmental particles with self-orienting capability is developed, where a single bubble is enclosed at the top of the super-segmented architecture. Such bubbles, driven by potential energy minimization, cause the particles to have a bubble-upward preferred orientation in liquid, enabling efficient decoding of their high-density signals in an interference-resistant manner. The particle preparation involves bubble encapsulation via the impact of a multicompartmental droplet on the liquid surface and overall stabilization via rational crosslinking. The conditions for obtaining these particles are systematically investigated. Methodological compatibility with materials is demonstrated by different hydrogel particles. Finally, by encapsulating cargoes of interest, these particles have found broad applications in actuators, multiplexed detection, barcodes, and multicellular systems.

Stability of the Arginine-Ornithine-Citrulline Cycle Maintained by Tumor-Stroma Interactions in Cell Coculture Hydrogel Microspheres.

Tumor-stroma interaction is the core process of tumor metastasis. Therefore, building a model of tumor-stromal cell communication is crucial for understanding the tumor metastasis process and curing cancer completely. In this research, a controllable three-dimensional (3D) tumor-stroma coculture microsphere model based on droplet microfluidic technology was developed to culture human lung cancer cells (A549 cell) and fibroblast cells (NIH-3T3 cell) using core-shell hydrogel microspheres to partition different kinds of cells. In our coculture model, tumor cells show a trend of epithelial-mesenchymal transition (EMT): a decrease in the number of surface E-cadherin and an increase in the number of N-cadherin. At the same time, fibroblasts are activated into cancer-associated fibroblasts (CAFs) as the level of interleukin-6 (IL-6) released is increased. In addition, an interesting phenomenon was discovered; in the absence of fibroblasts, the metabolism of the tumor cell culture alone leads to arginine depletion and citrulline accumulation, whereas a coculture can maintain the arginine-ornithine-citrulline cycle to reach equilibrium after 72 h, and the balance increases the stress resistance of tumor cells. This discovery may provide a new direction for understanding tumor resistance.

Multicompartmental Hydrogel Microspheres as a Tool for Multicomponent Analysis.

Developing advanced tools for multicomponent analysis is an open challenge in engineering and life science. Herein, multicompartmental hydrogel microspheres with multi-material compatibility and structural scalability are developed as a tool for multicomponent analysis at a single-particle level. Microfluidic technology endows particles with adjustable sizes and super-segmented layouts that can be used to load various analyte probes. In order to perform multicomponent analysis, these microspheres are structurally divided into identifier regions for indicating reading direction and analyte regions for detecting target molecules. The multiplex detection ability of these particles is demonstrated in microRNA bioassays with high specificity and sensitivity. The multi-target analysis is performed on a single-particle level, and the bioassay is free of conventional labeling interference. We expect these particles to reach their potential in clinical diagnostics.

The preventive effect of dexmedetomidine on anesthesia complications in strabismus surgery: a systematic review and meta-analysis.

OBJECTIVE: Dexmedetomidine is a medication that has analgesic, sedative, and anti-anxiety properties. In the clinical, it is often used to prevent common complications associated with strabismus surgery, including postoperative delirium, postoperative nausea and vomiting, postoperative pain, and oculocardiac reflex. However, its effectiveness and side effects of the present studies are different. The sample sizes of the present studies on the prevention of complications of dexmedetomidine are small. Therefore, this study evaluates the efficacy of dexmedetomidine in preventing anesthesia-related complications in strabismus surgery through a systematic review and meta-analysis. METHODS: Literature was retrieved from 10 commonly used databases and randomized controlled trials published up to May 2022 were sought. The included studies compared the intervention effects of dexmedetomidine versus placebo on anesthesia-related complications in surgery. The occurrence rates of postoperative delirium, postoperative nausea and vomiting, postoperative pain, and oculocardiac reflex in patients undergoing strabismus surgery were evaluated. Statistical analyses and forest plots were generated using Review Manager and STATA software. Binary outcomes were measured using relative risk (RR) with a 95% confidence interval for each outcome. The Cochrane risk of bias tool was used to assess the bias and risk in the studies that met the inclusion criteria. RESULTS: A total of 13 articles were ultimately included in the analysis, comprising 1,018 patients who underwent strabismus surgery. The dexmedetomidine group, compared to the placebo group, demonstrated significant reductions in the incidence of postoperative delirium (RR = 0.73, P = 0.001), severe postoperative delirium (RR = 0.45, P = 0.005), postoperative nausea and vomiting (RR = 0.48, P < 0.0001), and the need for supplemental analgesia postoperatively (RR = 0.60, P = 0.004). Additionally, subgroup analysis revealed that intravenous administration of dexmedetomidine significantly reduced the incidence of oculocardiac reflex (RR = 0.50, P = 0.001). In contrast, intranasal administration of dexmedetomidine did not have a significant effect on the incidence of oculocardiac reflex (RR = 1.22, P = 0.15). There was a significant difference between the subgroups (P = 0.0005, I2 = 91.7%). CONCLUSION: Among patients undergoing strabismus surgery, the use of dexmedetomidine can alleviate postoperative delirium and reduce the incidence of postoperative nausea and vomiting, as well as postoperative pain. Moreover, intravenous administration of dexmedetomidine can lower the occurrence rate of the oculocardiac reflex.

Irisin reduces the abnormal reproductive and metabolic phenotypes of PCOS by regulating the activity of brown adipose tissue in mice†.

Polycystic ovary syndrome (PCOS) is a common endocrine and metabolic disease in women, with clinical manifestations of anovulation and hyperandrogenaemia. The treatment of PCOS mainly focuses on improving clinical symptoms, such as insulin sensitivity or menstrual disorder, through drug treatment. However, due to the pathogenesis diversity of PCOS, there is still a lack of effective treatment in clinics. Metabolic disorder is the key factor in the occurrence of PCOS. Brown adipose tissue (BAT) is a special adipose tissue in the human body that can participate in metabolic balance by improving heat production. BAT has been demonstrated to be an important substance involved in the metabolic disorder of PCOS. Although increasing evidence indicates that BAT transplantation can improve the symptoms of PCOS, it is difficult to achieve BAT transplantation at present due to technical limitations. Stimulation of BAT activation by exogenous substances may be an effective alternative therapy for PCOS. In this study, we investigated the effects of Irisin on dehydroepiandrosterone (DHEA)-induced PCOS in mice and evaluated the effect of Irisin on serum hormone levels and changes in body temperature, body weight, and ovarian morphology. In our study, we found that Irisin can enhance the thermogenesis and insulin sensitivity of PCOS mice by activating the function of BAT. In addition, Irisin treatment can correct the menstrual cycle of PCOS mice, improve the serum steroid hormone disorder status, and reduce the formation of ovarian cystic follicles. In conclusion, our results showed that Irisin treatment significantly improved the metabolic disorder of PCOS and may provide a new and alternative therapy for the treatment of this pathology.

Global methane and nitrous oxide emissions from inland waters and estuaries.

Inland waters (rivers, reservoirs, lakes, ponds, streams) and estuaries are significant emitters of methane (CH4 ) and nitrous oxide (N2 O) to the atmosphere, while global estimates of these emissions have been hampered due to the lack of a worldwide comprehensive data set of CH4 and N2 O flux components. Here, we synthesize 2997 in-situ flux or concentration measurements of CH4 and N2 O from 277 peer-reviewed publications to estimate global CH4 and N2 O emissions from inland waters and estuaries. Inland waters including rivers, reservoirs, lakes, and streams together release 95.18 Tg CH4  year-1 (ebullition plus diffusion) and 1.48 Tg N2 O year-1 (diffusion) to the atmosphere, yielding an overall CO2 -equivalent emission total of 3.06 Pg CO2  year-1 . The estimate of CH4 and N2 O emissions represents roughly 60% of CO2 emissions (5.13 Pg CO2  year-1 ) from these four inland aquatic systems, among which lakes act as the largest emitter for both CH4 and N2 O. Ebullition showed as a dominant flux component of CH4 , contributing up to 62%-84% of total CH4 fluxes across all inland waters. Chamber-derived CH4 emission rates are significantly greater than those determined by diffusion model-based methods for commonly capturing of both diffusive and ebullitive fluxes. Water dissolved oxygen (DO) showed as a dominant factor among all variables to influence both CH4 (diffusive and ebullitive) and N2 O fluxes from inland waters. Our study reveals a major oversight in regional and global CH4 budgets from inland waters, caused by neglecting the dominant role of ebullition pathways in those emissions. The estimated indirect N2 O EF5 values suggest that a downward refinement is required in current IPCC default EF5 values for inland waters and estuaries. Our findings further indicate that a comprehensive understanding of the magnitude and patterns of CH4 and N2 O emissions from inland waters and estuaries is essential in defining the way of how these aquatic systems will shape our climate.

Development and validation of the Sources of Meaning in Life Scale for the Elderly in China.

OBJECTIVES: People who find meaning in life can endure 'any' pain. However, there were no tools to investigate elderly individuals' sources of meaning in life in China. This study aimed to develop the Sources of Meaning in Life Scale for the Elderly (SMSE), and examine the validation and reliability in Chinese elderly. METHODS: A 43-item pool of SMSE was formed by combining the preliminary interview and literature review. A cross-sectional survey of 627 elderly people was then conducted in two community health service centers, two hospitals, and two nursing homes in Guangzhou by the convenience sampling method. Test-retest reliability was assessed with 24 elderly persons. RESULTS: Six dimensions, containing family (four items), social support (four items), value (seven items), life security (four items), personal development (four items), and leisure activity (five items) explained 62.16% of the variance in total. Confirmatory factor analysis model fitting indices were χ2 = 694.652, df = 330, χ2/df = 2.105, SRMR = 0.0695, GFI = 0.853, IFI = 0.905, TLI = 0.889, CFI = 0.903, and RMSEA = 0.062. The Cronbach's alpha value of the scale was 0.924, while that of each dimension was between 0.727 and 0.870. The inter-class correlation (ICC) of the scale was 0.856. CONCLUSION: The SMSE has good reliability and validity that can be used to evaluate the sources of meaning and meaning in life for the elderly.

YY1-PVT1 affects trophoblast invasion and adhesion by regulating mTOR pathway-mediated autophagy.

Insufficient trophoblast invasion is the key factor for the occurrence of recurrent spontaneous abortions (RSA). Our previous studies identified Yin Yang 1 (YY1) as a transcription factor involved in the regulation of trophoblast invasiveness at the maternal-fetal interface. Long noncoding RNAs (lncRNAs) can regulate gene expression and autophagy in many ways. The purpose of this study was to explore the relationship between YY1 and lncRNAs and the mechanism by which lncRNAs affect the biological behavior of trophoblasts. Bioinformatic analysis predicted that YY1 had three binding sites in the plasmacytoma variant translocation 1 (PVT1) promoter region. Chromatin immunoprecipitation experiments and electrophoretic mobility shift assays verified that YY1 can directly bind to the PVT1 promoter. Compared with its expression levels in human placental villi tissue samples from the normal pregnancy group, the PVT1 expression levels were significantly lower in tissues from the RSA group. PVT1 knockdown significantly reduced adhesion, invasion, autophagy, and mTOR expression in HTR-8/SVneo cells and greatly increased apoptosis in vitro. This study revealed a novel regulatory pathway in which YY1 can act directly on PVT1 promoter to regulate its transcription, which further affects trophoblast invasion and adhesion by regulating autophagy via the mTOR pathway, and these effects might be involved in RSA pathogenesis.

Increased soil release of greenhouse gases shrinks terrestrial carbon uptake enhancement under warming.

Warming can accelerate the decomposition of soil organic matter and stimulate the release of soil greenhouse gases (GHGs), but to what extent soil release of methane (CH4 ) and nitrous oxide (N2 O) may contribute to soil C loss for driving climate change under warming remains unresolved. By synthesizing 1,845 measurements from 164 peer-reviewed publications, we show that around 1.5°C (1.16-2.01°C) of experimental warming significantly stimulates soil respiration by 12.9%, N2 O emissions by 35.2%, CH4 emissions by 23.4% from rice paddies, and by 37.5% from natural wetlands. Rising temperature increases CH4 uptake of upland soils by 13.8%. Warming-enhanced emission of soil CH4 and N2 O corresponds to an overall source strength of 1.19, 1.84, and 3.12 Pg CO2 -equivalent/year under 1°C, 1.5°C, and 2°C warming scenarios, respectively, interacting with soil C loss of 1.60 Pg CO2 /year in terms of contribution to climate change. The warming-induced rise in soil CH4 and N2 O emissions (1.84 Pg CO2 -equivalent/year) could reduce mitigation potential of terrestrial net ecosystem production by 8.3% (NEP, 22.25 Pg CO2 /year) under warming. Soil respiration and CH4 release are intensified following the mean warming threshold of 1.5°C scenario, as compared to soil CH4 uptake and N2 O release with a reduced and less positive response, respectively. Soil C loss increases to a larger extent under soil warming than under canopy air warming. Warming-raised emission of soil GHG increases with the intensity of temperature rise but decreases with the extension of experimental duration. This synthesis takes the lead to quantify the ecosystem C and N cycling in response to warming and advances our capacity to predict terrestrial feedback to climate change under projected warming scenarios.

CTLA4 has a profound impact on the landscape of tumor-infiltrating lymphocytes with a high prognosis value in clear cell renal cell carcinoma (ccRCC).

Background: Cytotoxic T-lymphocyte associated protein 4 (CTLA4) inhibitors have been shown to significantly prolong the overall survival (OS) in a wide range of cancers. However, its application in clear cell renal cell carcinoma (ccRCC) is limited due to the therapy response, and the prognostic value of CTLA4 in ccRCC has not been investigated in detail. Methods: By using immunohistochemistry, Kaplan-Meier (K-M) analysis, uni- and multi-variate Cox analysis, we comprehensively and systematically studied the prognostic value of CTLA4 in ccRCC. Then, we applied Gene Ontology (GO), the Kyoto Encyclopedia of Genes and Genomes (KEGG) and CIBERSORT, ESTIMATE algorithm, ssGSEA and somatic mutation analyses to reveal the impact of CTLA4 on the landscape of tumor-infiltrating lymphocytes (TILs) infiltration and genetic mutation. Besides, given current concerns caused by combined immunotherapy, we also investigated the relationship between CTLA4 and other immune checkpoints. Results: In vitro experiment and data mining showed that, CTLA4 was up-regulated in ccRCC tissues and closely related to the disease progression as well as a poor prognosis. Deeper researches demonstrated that CTLA4 regulates T cell activation and was significantly linked to TIL-abundant tumor microenvironment (TME), but was accompanied by an immunosuppressed phenotype. Mutation analysis showed that CTLA4 was associated with more frequent BRCA-associated protein 1 (BAP1) mutation. Moreover, we found that CTLA4 was markedly correlated with multiple immune checkpoints, which suggested that ccRCC patients with high expressed CTLA4 may benefit more from immune checkpoint blockades (ICBs) combined therapy. Conclusion: CTLA4 has a profound impact on the landscape of TILs and genetic mutation, and can be used as the biomarker with high prognosis value in ccRCC.

Controllable Synthesis of Multicompartmental Particles Using 3D Microfluidics.

A microfluidic assembly method based on a microfluidic chip and capillary device was developed to create multicompartmental particles. The microfluidic chip design endows the particles with regulable internal structure. By adjusting the microstructure of the chip, the diameter of the capillary, the gap length between the two microfluidic components, and the flow rates, the size of the particles and the number or the ratio of different regions within the particle could be widely varied. As a proof of concept, we have produced some complicated particles that even contain 20 compartments. Furthermore, the potential applications of the anisotropic particles are explored by encapsulating magnetic beads, fluorescent nanoparticles, and the cells into different compartments of the microparticles. We believe that this method will open new avenues for the design and application of multicompartmental particles.

Multifunctional Regulation of 3D Cell-Laden Microsphere Culture on an Integrated Microfluidic Device.

Three-dimensional (3D) hydrogel microspheres have aroused increasing attention as an in vitro cell culture model. Yet the preservation of cells' original biological properties has been overlooked during model construction. Here we present an integrated microfluidic device to accomplish the overall process including cell-laden microsphere generation, online extraction, and dynamic-culture. The method extends the noninvasive and nonsuppression capabilities of the droplet preparation system and provides a constant microenvironment, which reduces intracellular oxidative stress damage and the accumulation of mitochondria. Compared to the conventional preparation method, the coculture model of tumor-endothelial construction on an integrated platform displays high-level angiogenic protein expression. We believe that this versatile and biocompatible platform will provide a more reliable analysis tool for tissue engineering and cancer therapy.

2-Deoxy-D-Glucose Modified Magnetic Nanoparticles with Dual Functional Properties: Nanothermotherapy and Magnetic Resonance Imaging.

Superparamagnetic iron oxide nanoparticles (SPIONs) with appropriate surface chemistry have attracted wild attention in medical and biological application because of their current and potential usefulness such as magnetic resonance imaging (MRI) contrast enhancement, magnetic mediated hyperthermia (MMH), immunoassay, and in drug delivery, etc. In this study, we investigated the MRI contrast agents and MMH mediators properties of the novel 2-deoxy-D-glucose (2-DG) modified SPIONs. As a non-metabolizable glucose analogue, 2-DG can block glycolysis and inhibits protein glycosylation. Moreover, SPIONs coated with 2-DG molecules can be particularly attractive to resource-hungry cancer cells, therefore to realize the targeting strategy for the SPIONs. SPIONs with amino silane as the capping agent for amino-group surface modification were synthesized by the chemical co-precipitation method with modification. Glutaraldehyde was further applied as an activation agent through which 2-DG was conjugated to the amino-coated SPIONs. Physicochemical characterizations of the 2-DG-SPIONs, such as surface morphology, surface charge and magnetic properties were investigated by Transmission Electron Microscopy (TEM), ζ-Potential and Vibrating Sample Magnetometer (VSM), etc. Magnetic inductive heating characteristics of the 2-DG-SPIONs were analyzed by exposing the SPIONs suspension (magnetic fluid) under alternative magnetic field (AMF). U-251 human glioma cells with expression of glucose transport proteins type 1 and 3 (GLUT1 and GLUT 3), and L929 murine fibroblast cell as negative control, were employed to study the effect of 2-DG modification on the cell uptake for SPIONs. TEM images for ultra-thin sections as well as ICP-MS were applied to evaluate the SPIONs internalization within the cells. In vitro MRI was performed after cells were co-incubated with SPIONs and the T2 relaxation time was measured and compared. The results demonstrate that 2-DG-SPIONs were supermagnetic and in spherical shape with -10 nm diameter. Possessing ideal magnetic inductive heating characteristics, which can generate very rapid and efficient heating while upon AMF exposure, 2-DG-SPIONs can be applied as novel candidature of magnetic nanothermotherapy for cancer treatment. Modification of 2-DG can greatly promote the cell uptake of SPIONs and such cellular uptake of 2-DG-SPIONs was time dependent. Surface coating by 2-DG can remarkably enhance the MR imaging ability for the SPIONs on the cells of U251 cancer cells. In summary, our investigation provides a novel glucose analogue modified SPIONs with potential application in the targeting cancer nanothermotherapy and MR imaging.

An optofluidic system for the concentration gradient screening of oocyte protection drugs.

Oocytes protective drug screening is essential for the treatment of reproductive diseases. However, few studies construct the oocyte in vitro drug screening microfluidic systems because of their enormous size, scarcity, and sensitivity to the culture environment. Here, we present an optofluidic system for oocyte drug screening and state analysis. The system consists of two parts: an open-top drug screening microfluidic chip and an optical Fourier filter analysis part. The open-top microfluidic chip anchors single oocyte with hydrogel and allows nutrient and gas environment updating which is essential for oocyte culturing. The optical filter analysis part is used to accurately analyse the status of oocytes. Based on this system, we found that fluorene-9-bisphenol (BHPF) damaged the oocyte spindle in a dose-dependent manner, a high dose of melatonin (10-3 M) effectively reduces the percentage of abnormally arranged chromosomes of oocytes exposed to 40 µM BHPF. This optofluidic system shows great promise for the culture of oocytes and demonstrates the robust ability for convenient multi-concentration oocytes drug screening. This technology may benefit further biomedicine and reproductive toxicology applications in the lab on a chip community.

Capture the Moment: High-Speed Imaging With Spiking Cameras Through Short-Term Plasticity.

High-speed imaging can help us understand some phenomena that are too fast to be captured by our eyes. Although ultra-fast frame-based cameras (e.g., Phantom) can record millions of fps at reduced resolution, they are too expensive to be widely used. Recently, a retina-inspired vision sensor, spiking camera, has been developed to record external information at 40, 000 Hz. The spiking camera uses the asynchronous binary spike streams to represent visual information. Despite this, how to reconstruct dynamic scenes from asynchronous spikes remains challenging. In this paper, we introduce novel high-speed image reconstruction models based on the short-term plasticity (STP) mechanism of the brain, termed TFSTP and TFMDSTP. We first derive the relationship between states of STP and spike patterns. Then, in TFSTP, by setting up the STP model at each pixel, the scene radiance can be inferred by the states of the models. In TFMDSTP, we use the STP to distinguish the moving and stationary regions, and then use two sets of STP models to reconstruct them respectively. In addition, we present a strategy for correcting error spikes. Experimental results show that the STP-based reconstruction methods can effectively reduce noise with less computing time, and achieve the best performances on both real-world and simulated datasets.

Microfluidic Engineering of Crater-Terrain Hydrogel Microparticles: Toward Novel Cell Carriers.

Fabrication and application of novel anisotropic microparticles are of wide interest. Herein, a new method for producing novel crater-terrain hydrogel microparticles is presented using a concept of droplet-aerosol impact and regional polymerization. The surface pattern of microparticles is similar to the widespread "crater" texture on the lunar surface and can be regulated by the impact morphology of aerosols on the droplet surface. Methodological applicability was demonstrated by producing ionic-cross-linked (alginate) and photo-cross-linked (poly(ethylene glycol) diacrylate, PEGDA) microparticles. Additionally, the crater-terrain microparticles (CTMs) can induce nonspecific protein absorption on their surface to acquire cell affinity, and they were exploited as cell carriers to load living cells. Cells could adhere and proliferate, and a special cellular adhesion fingerprint was observed on the novel cell carrier. Therefore, the scalable manufacturing method and biological potential make the engineered microparticles promising to open a new avenue for exploring cell-biomaterial crosstalk.

In situ polymerized ionic liquids in polyester fiber composite membranes for detection of trace oil.

In situ trace detection on ultra-clean surfaces is an important technology. The polyester fiber (PF) was introduced to serve as the template, to which the ionic liquids were bonded by hydrogen bonding. Polymerized ionic liquids (PIL) in PF were formed by in situ polymerization with the azodiisobutyronitrile (AIBN) and IL. The trace oil on metal surfaces was enriched by the composite membrane based on similar compatibility principle. The absolute recovery of the trace oil ranged from 91%-99% using this composite membrane. In the extraction samples, desirable linear correlations were obtained for trace oil in the range of 1.25-20 mg/mL. It has been proven that a 1 cm2 PIL-PF composite membrane can effectively extract as little as 1 mg of lubricating oil on an ultra-clean metal surface of 0.1 m2 with the LOD of 0.9 mg/mL, making it a promising material for in situ detection of trace oil on metal surfaces.

Multidimensional controllable fabrication of tumor spheroids based on a microfluidic device.

Multicellular tumor spheroids (MCTSs) are in vitro solid tumor models with physiological relevance. To achieve robust process control, a MCTS fabrication method that combines cell membrane engineering and droplet microfluidic techniques is designed. The fluidic control and the chemical interactions between biotin and streptavidin enable artificial cell aggregation to be accomplished in seconds. Then, spheroids with a uniform size are fabricated within alginate microcapsules. Microfluidic mixing-based cell aggregation regulates the cell aggregate size and the spheroid composition, and the microcapsules regulate the size of spheroids from 120 to 180 µm. The method shows applicability for various cancer cell lines, including HCT116, HepG2, and A549. In addition, composite colon cancer spheroids consisting of HCT116 and NIH3T3 with predetermined cell ratios and uniform distributions are produced. The generated MCTSs are assessed using the ELISA and UPLC-MS/MS techniques. The release of vascular endothelial growth factor (VEGF) and the 5-fluorouracil (5-FU) resistance differ in the monotypic and cocultured colon cancer models. Our method provides a robust way to produce consistent and customized MCTSs in cancer research and drug screening.

Multivariate Porous Aromatic Frameworks with High Porosity and Hierarchical Structures for Enzyme Immobilization.

As materials with permanently porous structures and readily modifying availability, porous aromatic frameworks (PAFs) are considered as promising porous materials with versatile functionality. Currently the designable synthesis of PAFs with the desired surface area and pore size is still a challenge, and instead kinetically irreversible coupling reactions for PAFs synthesis has resulted in the unpredictable connection of building units. Herein, a series of PAFs with highly porous and hierarchical structures were successfully synthesized through a multivariate inspired strategy, where multiple building units with various topologies and sizes were selected for PAFs synthesis. All the PAFs synthesized through this strategy possessed hierarchical structures and high specific surface areas at the same time. Encouraged by their high surface area and hierarchical structures, we loaded lipase onto one of the multivariate PAFs. The enzyme loading content of the obtained lipase@PAF-147 was as high as 1456 mg g-1, which surpassed any other currently reported enzyme loading materials. The lipase@PAF-147 also exhibited favorable catalytic activity and stability to a model reaction of p-nitrophenyl caprylate (p-NPC) hydrolysis. This multivariate strategy inspired synthetic method broadens the selection of building units for PAFs design and opens a new avenue for the design of functional porous materials.