Diseased Erythrocyte Enrichment Based on I-Shaped Pillar DLD Arrays.

Enrichment of erythrocytes is a necessary step in the diagnosis of blood diseases. Due to the high deformability and viscosity of erythrocytes, they cannot be regarded as stable point-like solids, so the influence of their deformability on fluid dynamics must be considered. Therefore, by using the special effect of an I-shaped pillar (I-pillar) on erythrocytes, erythrocytes with different deformability can be made to produce different provisional distances in the chip, so as to achieve the separation of the two kinds of erythrocytes. In this study, a microfluidic chip was designed to conduct a control test between erythrocytes stored for a long time and fresh erythrocytes. At a specific flow rate, the different deformable erythrocytes in the chip move in different paths. Then, the influence of erythrocyte deformability on its movement trajectory was analyzed by two-dimensional finite element flow simulation. DLD sorting technology provides a new method for the sorting and enrichment of diseased erythrocytes.

Numerical Studies on the Motions of Magnetically Tagged Cells Driven by a Micromagnetic Matrix.

Precisely controlling magnetically tagged cells in a complex environment is crucial to constructing a magneto-microfluidic platform. We propose a two-dimensional model for capturing magnetic beads from non-magnetic fluids under a micromagnetic matrix. A qualitative description of the relationship between the capture trajectory and the micromagnetic matrix with an alternating polarity configuration was obtained by computing the force curve of the magnetic particles. Three stages comprise the capture process: the first, where motion is a parabolic fall in weak fields; the second, where the motion becomes unpredictable due to the competition between gravity and magnetic force; and the third, where the micromagnetic matrix finally captures cells. Since it is not always obvious how many particles are adhered to the surface, attachment density is utilized to illustrate how the quantity of particles influences the capture path. The longitudinal magnetic load is calculated to measure the acquisition efficiency. The optimal adhesion density is 13%, and the maximum adhesion density is 18%. It has been demonstrated that a magnetic ring model with 100% adhesion density can impede the capture process. The results offer a theoretical foundation for enhancing the effectiveness of rare cell capture in practical applications.

Rapid Production of Nanoscale Liposomes Using a 3D-Printed Reactor-In-A-Centrifuge: Formulation, Characterisation, and Super-Resolution Imaging.

Nanoscale liposomes have been extensively researched and employed clinically for the delivery of biologically active compounds, including chemotherapy drugs and vaccines, offering improved pharmacokinetic behaviour and therapeutic outcomes. Traditional laboratory-scale production methods often suffer from limited control over liposome properties (e.g., size and lamellarity) and rely on laborious multistep procedures, which may limit pre-clinical research developments and innovation in this area. The widespread adoption of alternative, more controllable microfluidic-based methods is often hindered by complexities and costs associated with device manufacturing and operation, as well as the short device lifetime and the relatively low liposome production rates in some cases. In this study, we demonstrated the production of liposomes comprising therapeutically relevant lipid formulations, using a cost-effective 3D-printed reactor-in-a-centrifuge (RIAC) device. By adjusting formulation- and production-related parameters, including the concentration of polyethylene glycol (PEG), temperature, centrifugation time and speed, and lipid concentration, the mean size of the produced liposomes could be tuned in the range of 140 to 200 nm. By combining selected experimental parameters, the method was capable of producing liposomes with a therapeutically relevant mean size of ~174 nm with narrow size distribution (polydispersity index, PDI ~0.1) at a production rate of >8 mg/min. The flow-through method proposed in this study has potential to become an effective and versatile laboratory-scale approach to simplify the synthesis of therapeutic liposomal formulations.

Evaporation issues of acoustically levitated fuel droplets.

Fuel droplet evaporation is essential to the generation of flammable mixtures in thermal engines. Generally, liquid fuel is injected directly into the hot, high-pressure atmosphere to form scattered droplets. Many investigations on droplet evaporation have been conducted with techniques involving the influence of boundaries, such as suspended wires. Ultrasonic levitation is a non-contact and non-destructive technology that can avoid the impact of hanging wire on droplet shape and heat transfer. Besides, it can simultaneously levitate multiple droplets and allow them to associate with each other or be used to study droplet instability behaviors. This paper reviews the influences of the acoustic field on levitated droplets, the evaporation characteristics of acoustically levitated droplets, and the prospects and limitations of ultrasonic suspension methods for droplet evaporation, which can serve as references for relevant studies.

Photodynamic therapy: a new approach to the treatment of Nontuberculous Mycobacterial skin and soft tissue infections.

Nontuberculous mycobacterial skin and soft tissue infections are rising and are causing social concern due to the growth of cosmetic dermatology and immune-compromised populations. For the treatment of nontuberculous mycobacteria, several novel strategies have been investigated. One of them, photodynamic therapy, is a recently developed therapeutic strategy that has shown promise in managing nontuberculous mycobacterial skin and soft tissue infections. In this review, we first present an overview of the current status of the therapy and then summarize and analyze the cases of photodynamic therapy used to treat nontuberculous mycobacterial skin and soft tissue infections. We also discussed the feasibility of photodynamic therapy for treating nontuberculous mycobacterial skin soft tissue infections and the related mechanisms, providing a potential new option for clinical treatment.

Mitophagy associated self-degradation of phosphorylated MAP4 guarantees the migration and proliferation responses of keratinocytes to hypoxia.

Our previous study has announced that phosphorylated microtubule-associated protein 4 (p-MAP4) accelerated keratinocytes migration and proliferation under hypoxia through depolymerizing microtubules. However, p-MAP4 should exhibit inhibitory effects on wound healing, for it also impaired mitochondria. Thus, figuring out the outcome of p-MAP4 after it impaired mitochondria and how the outcome influenced wound healing were far-reaching significance. Herein, the results revealed that p-MAP4 might undergo self-degradation through autophagy in hypoxic keratinocytes. Next, p-MAP4 activated mitophagy which was unobstructed and was also the principal pathway of its self-degradation triggered by hypoxia. Moreover, both Bcl-2 homology 3 (BH3) and LC3 interacting region (LIR) domains had been verified in MAP4, and they endowed MAP4 with the capability to synchronously function as a mitophagy initiator and a mitophagy substrate receptor. And, mutating any one of them ruined hypoxia-induced self-degradation of p-MAP4, resulting in destroyed proliferation and migration responses of keratinocytes to hypoxia. Our findings unviewed that p-MAP4 experienced mitophagy-associated self-degradation through utilizing its BH3 and LIR domains under hypoxia. As a result, the mitophagy-associated self-degradation of p-MAP4 guaranteed the migration and proliferation responses of keratinocytes to hypoxia. Together, this research provided a bran-new pattern of proteins in regulating wound healing, and offered a new direction for intervening wound healing.

Local CRF and oxytocin receptors correlate with female experience-driven avoidance change and hippocampal neuronal plasticity.

Understanding how experiences affect females' behaviors and neuronal plasticity is essential for uncovering the mechanism of neurodevelopmental disorders. The study explored how neonatal maternal deprivation (MD) and post-weaning environmental enrichment (EE) impacted the CA1 and DG's neuronal plasticity in the dorsal hippocampus, and its relationships with passive avoidance, local corticotrophin-releasing factor (CRF) levels, and oxytocin receptor (OTR) levels in female BALB/c mice. The results showed that MD damaged passive avoidance induced by foot shock and hotness, and EE restored it partially. In the CA1, MD raised CRF levels and OTR levels. Parallelly, MD increased synaptic connection levels but reduced the branches' numbers of pyramidal neurons. Meanwhile, in the DG, MD increased OTR levels but lowered CRF levels, DNA levels, and spine densities. EE did not change the CA1 and DG's CRF and OTR levels. However, EE added DG's dendrites of granular cells. The additive of MD and EE raised CA1's synaptophysin and DG's postsynaptic density protein-95 and OTR levels, and meanwhile, shaped avoidance behaviors primarily similar to the control. The results suggest that experience-driven avoidance change and hippocampal neuronal plasticity are associated with local CRF and OTR levels in female mice.

In vitro biocompatibility study of EDC/NHS cross-linked silk fibroin scaffold with olfactory ensheathing cells.

In this paper, we investigated silk fibroin (SF) cross-linked by 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide/N-hydroxysuccinimide (EDC/NHS) and its biocompatibility with olfactory ensheathing cells (OECs). After cross-linked with different concentrations of EDC/NHS solutions, SF scaffolds were analyzed by different techniques such as scanning electron microscopy, Fourier transform infrared spectra, x-ray diffraction, tensile machine and water contact angle assay. As to their structures, we found 4.5% EDC/NHS cross-linked SF possessed a more significant increase of ß-sheet and a decrease of α-helix than 1.5% group. These changes helped SF achieve excellent mechanical properties. While more remarkable improvement of hydrophilicity was seen in 1.5% EDC/NHS treated SF. Immunofluorescence, MTT, Annexin-V/PI and ELISA analysis were then conducted to determine the states and functions of OECs on the scaffolds. OECs on 4.5% EDC/NHS cross-linked SF seemed insufficient to spread, and the proliferation was limited on 4 and 6 days. Moreover, 4.5% EDC/NHS exerted adverse effects on cell survival and nerve growth factor (NGF) secretion at day 4, but not 1.5% EDC/NHS. Taken together, SF scaffolds showed improved physical and hydrophilic properties through cross-linking. 1.5% EDC/NHS cross-linked SF scaffolds showed significant advantages between mechanical property and the states and functions with OECs, which has the potential to be used for neural repairing.

Reciprocating Oscillation of a Floating Ferrofluid Marble Triggered by Magnetic Fields.

Liquid marbles have the potential for microfluidic transport, medical diagnostics, and chemical analysis due to their negligible stickiness, environmental independence, and excellent mobility. Here, we report a non-contact manipulation strategy to arouse a reciprocating oscillation of ferrofluid marbles floating on the water surface, which can be used as microreactors. We experimentally investigated the quantitative relationship between the oscillation behavior, the applied magnetic field parameters, and the field regulation mechanism. The variables, including the magnetic field strength, marble volume, and switching period, are vital in determining the final state. The oscillation can be separated into three stages: transitional movement, compressive deformation, and rebound, before entering the next cycle. Accordingly, we created a manipulation technique for improving the mixing of inner reactants inside this marble container by remote-controlled shaking after optimizing with an oscillation model.

Eu3+-Doped Electrospun Polyvinylidene Fluoride-Hexafluoropropylene/Graphene Oxide Multilayer Composite Nanofiber for the Fabrication of Flexible Pressure Sensors.

The development of flexible materials with higher piezoelectric properties and electrostrictive response is of great significance in many applications such as wearable functional devices, flexible sensors, and actuators. In this study, we report an efficient fabrication strategy to construct a highly sensitive (0.72 kPa-1), red light-emitting flexible pressure sensor using electrospun Eu3+-doped polyvinylidene fluoride-hexafluoropropylene/graphene oxide composite nanofibers using a layer-by-layer technology. The high ß-phase concentration (96.3%) was achieved from the Eu3+-doped P(VDF-HFP)/GO nanofibers, leading to a high piezoelectricity of the composite nanofibers. We observed that a pressure sensor is enabled to generate an output voltage of 4.5 V. Furthermore, Eu3+-doped P(VDF-HFP)/GO composite nanofiber-based pressure sensors can also be used as an actuator as it has a good electrostrictive effect. At the same time, the nanofiber membrane has excellent ferroelectric properties and good fluorescence properties. These results indicate that this material has great application potential in the fields of photoluminescent fabrics, flexible sensors, soft actuators, and energy storage devices.

Dermal Fibroblast Migration and Proliferation Upon Wounding or Lipopolysaccharide Exposure is Mediated by Stathmin.

The dermal fibroblast is a crucial executor involved in wound healing, and lipopolysaccharide is a key factor in initiating the migration and proliferation of the dermal fibroblasts, followed by wound healing. However, the underlying molecular mechanism is still unknown. In this study, we demonstrated that stathmin increased concomitantly with p38/MAPK pathway activation by lipopolysaccharide stimulation of the human dermal fibroblast (HDF), which induced microtubule (MT) depolymerization followed by increased HDF migration and proliferation. In contrast, the application of taxol, the small interfering RNA transfection of stathmin, or the application of the p38/MAPK inhibitor SB203580 suppressed MT depolymerization and HDF migration and proliferation. Additionally, the overexpression of a MKK6(Glu) mutant, which constitutively activated p38/MAPK, resulted in MT depolymerization and, subsequently, promoted HDF migration and proliferation. Our data reveal a crucial role of stathmin in HDF migration and proliferation. These findings will provide new targets and strategies for clinical interventions in wound healing.

Non-Contact Monitoring on the Flow Status inside a Pulsating Heat Pipe.

The paper presents a concept of thermal-to-electrical energy conversion by using the oscillatory motion of magnetic fluid slugs which has potential to be applied in the field of sensors. A pulsating heat pipe (PHP) is introduced to produce vapor-magnetic fluid plug-slug flow in a snake-shaped capillary tube. As the magnetic fluid is magnetized by the permanent magnet, the slugs of magnetic fluid passing through the copper coils make the magnetic flux vary and produce the electromotive force. The peak values of induced voltage observed in our tests are from 0.1 mV to 4.4 mV. The effects of the slug velocity, heat input and magnetic particle volume concentration on the electromotive force are discussed. Furthermore, a theoretical model considering the fluid velocity of the working fluid, the inner radius of the PHP and the contact angle between the working fluid and the pipe wall is established. At the same time, the theoretical and experimental results are compared, and the influences of tube inner radius, working fluid velocity and contact angle on the induced electromotive force are analyzed.

Penetrable Nanoplatform for "Cold" Tumor Immune Microenvironment Reeducation.

Lack of tumor-infiltration lymphocytes (TILs) and resistances by overexpressed immunosuppressive cells (principally, myeloid-derived suppressor cells (MDSCs)) in tumor milieu are two major challenges hindering the effectiveness of immunotherapy for "immune-cold" tumors. In addition, the natural physical barrier existing in solid cancer also limits deeper delivery of drugs. Here, a tumor-targeting and light-responsive-penetrable nanoplatform (Apt/PDGs/@pMOF) is developed to elicit intratumoral infiltration of cytotoxic T cells (CTLs) and reeducate immunosuppressive microenvironment simultaneously. In particular, porphyrinic metal-organic framework (pMOF)-based photodynamic therapy (PDT) induces tumor immunogenic cell death (ICD) to promote CTLs intratumoral infiltration and hot "immune-cold" tumor. Upon being triggered by PDT, the nearly 10 nm adsorbed drug-loaded dendrimer de-shields from the nanoplatform and spreads into the deeper tumor, eliminating MDSCs and reversing immunosuppression, eventually reinforcing immune response. Meanwhile, the designed nanoplatform also has a systemic MDSC inhibition effect and moderate improvement of overall antitumor immune responses, resulting in effective suppression of distal tumors within less significant immune-related adverse effects (irAEs) induced.

The RCAN1.4-calcineurin/NFAT signaling pathway is essential for hypoxic adaption of intervertebral discs.

Calcipressin-1, also known as regulator of calcineurin 1 (RCAN1), can specifically bind calcineurin at or near the calcineurin A catalytic domain and downregulate calcineurin activity. However, whether RCAN1 affects the hypoxic intervertebral disc (IVD) phenotype through the calcineurin/NFAT signaling pathway remains unclear. First, we confirmed the characteristics of the degenerative nucleus pulposus (NP) by H&E, safranin O/fast green and Alcian blue staining, and detected increased RCAN1 levels in the degenerative NP by immunohistochemistry. Then, we demonstrated that the protein level of RCAN1.4 was higher than that of RCAN1.1 and progressively elevated from the control group to the Pfirrmann grade V group. In vitro, both hypoxia (1% O2) and overexpression of HIF-1α reduced the protein level of RCAN1.4 in rat NP cells in a dose- and time-dependent manner. We further found that miRNA-124, through a nondegradative pathway (without the proteasome or lysosome), suppressed the expression of RCAN1.4. As expected, calcineurin in NP cells was activated and primarily promoted nuclear translocation of NFATc1 under hypoxia or RCAN1.4 siRNA transfection. Furthermore, SOX9, type II collagen and MMP13 were elevated under hypoxia, RCAN1.4 siRNA transfection or NFATc1 overexpression. Using chromatin immunoprecipitation (ChIP) and a luciferase reporter assay (with mutation), we clarified that NFATc1 increasingly bound the SOX9 promotor region (bp -367~-357). Interaction of HIF-1α and NFATc1 promoted MMP13 transcription. Finally, we found that FK506 reversed hypoxia-induced activation of the calcineurin/NFAT signaling pathway in NP cells and an ex vivo model. Together, these findings show that the RCAN1.4-calcineurin/NFAT signaling pathway has a vital role in the hypoxic phenotype of NP cells. RCAN1.4 might be a therapeutic target for degenerative disc diseases.

Magnetically driven microfluidics for isolation of circulating tumor cells.

Circulating tumor cells (CTCs) largely contribute to cancer metastasis and show potential prognostic significance in cancer isolation and detection. Miniaturization has progressed significantly in the last decade which in turn enabled the development of several microfluidic systems. The microfluidic systems offer a controlled microenvironment for studies of fundamental cell biology, resulting in the rapid development of microfluidic isolation of CTCs. Due to the inherent ability of magnets to provide forces at a distance, the technology of CTCs isolation based on the magnetophoresis mechanism has become a routine methodology. This historical review aims to introduce two principles of magnetic isolation and recent techniques, facilitating research in this field and providing alternatives for researchers in their study of magnetic isolation. Researchers intend to promote effective CTC isolation and analysis as well as active development of next-generation cancer treatment. The first part of this review summarizes the primary principles based on positive and negative magnetophoretic isolation and describes the metrics for isolation performance. The second part presents a detailed overview of the factors that affect the performance of CTC magnetic isolation, including the magnetic field sources, functionalized magnetic nanoparticles, magnetic fluids, and magnetically driven microfluidic systems.

Reoccurring discogenic low back pain (LBP) after discoblock treated by oblique lumbar interbody fusion (OLIF).

OBJECTIVE: To determine the efficacy of OLIF in the treatment of reoccurring discogenic low back pain (LBP) after discoblock METHODS: We included 108 patients with LBP that was suspected to be discogenic (such as high intensity zone, Schmorl's nodes, Modic changes Type I, etc.), from August 2015 to August 2017. All patients underwent discography, and patients whose LBP was confirmed to be discogenic received discoblock. Patients who had reoccurring pain after discoblock underwent OLIF. Perioperative parameters and complications were recorded. The VAS and Oswestry Disability Index (ODI) were assessed at preoperation, and 1 week and 1, 3, 6, and 12 months after the surgery. The fusion rate was evaluated. RESULTS: Of 108 patients, 89 were confirmed to have discogenic LBP, and 32/89 patients with reoccurring LBP pain after discoblock underwent OLIF. Twenty-eight patients were followed up for ≥ 1 year. The OLIF operation lasted for 92 ± 34 min. Blood loss during the operation was 48 ± 15 ml. The mean incision length was 3.0 ± 0.6 cm. The average length of stay was 4.8 ± 1.9 days. The VAS and ODI scores decreased from 8.1 ± 1.7 preoperatively to 0.9 ± 0.4, and from 71.2 ± 11.3 to 9.3 ± 3.1, 12 months postoperatively, respectively. The total incidence of complications was 15.6%, including 2 cases of cage subsidence, 2 cases of ipsilateral hip flexor weakness, and 1 case of ipsilateral anterior thigh pain. All symptoms relieved or disappeared during follow-up. The fusion rate was 96.9%. CONCLUSIONS: Reoccurring discogenic LBP after discoblock should be considered as a suitable group for treatment by OLIF.

Dandelion-Like Tailorable Nanoparticles for Tumor Microenvironment Modulation.

Tumor-associated macrophages (TAMs) constitute over 50% of the number of cells within the tumor, playing a major role in tumor progression and invasion. Remodeling the tumor immune microenvironment by modulating TAM polarization has been emerging as a new and promising therapeutic strategy. However, the high interstitial fluid pressure and dense extracellular matrix lead to insufficient penetration of nanosized therapies. To overcome this dilemma, an acid-triggered size-changeable nanoparticle (aptamer/acid sensitive linker crosslinked DGL/zoledronic acid, i.e., Apt@(DGL-ZA) n NPs) with effective tumor distribution, extravasation, and penetration is designed. Dendrigraft poly-L-lysines (DGLs) which can induce tumor autophagy as mimics of natural abnormal proteins are crosslinked via a mild-acid-responsive linker (1,6-bis(4-formylbenzoyloxy) hexane). Long circulation property and tumor penetration are achieved simultaneously by catching DGLs in neutral pH while releasing them in the tumor's pH, like dandelion seeds in midair. The macrophage conditioning agent zoledronic acid (ZA) is loaded on DGLs by the charge attraction. A Tenascin-C targeting aptamer (GBI-10) is modified onto (DGL-ZA) n NPs for a tumor-homing effect. Apt@(DGL-ZA) n NPs show both enhanced penetration in in vitro 3D triple negative breast cancer spheroids and in vivo tumor tissues. Effective macrophage regulation, enhanced tumor autophagy, and excellent in vivo antitumor efficacy are achieved, suggesting this tactic as a significant antitumor strategy.

Two-dimensional Simulation of Motion of Red Blood Cells with Deterministic Lateral Displacement Devices.

Deterministic lateral displacement (DLD) technology has great potential for the separation, enrichment, and sorting of red blood cells (RBCs). This paper presents a numerical simulation of the motion of RBCs using DLD devices with different pillar shapes and gap configurations. We studied the effect of the pillar shape, row shift, and pillar diameter on the performance of RBC separation. The numerical results show that the RBCs enter "displacement mode" under conditions of low row-shift (∆λ < 1.4 µm) and "zigzag mode" with large row shift (∆λ > 1.5 µm). RBCs can pass the pillar array when the size of the pillar (d > 6 µm) is larger than the cell size. We show that these conclusions can be helpful for the design of a reliable DLD microfluidic device for the separation of RBCs.