Hyaluronic acid-decorated curcumin-based coordination nanomedicine for enhancing the infected diabetic wound healing.

Persistent over-oxidation, inflammation and bacterial infection are the primary reasons for impaired wound repairing in diabetic patients. Therefore, crucial strategies to promote diabetic wound repairing involve suppressing the inflammatory response, inhibiting bacterial growth and decreasing reactive oxygen species (ROS) within the wound. In this work, we develop a multifunctional nanomedicine (HA@Cur/Cu) designed to facilitate the repairing process of diabetic wound. The findings demonstrated that the synthesized infinite coordination polymers (ICPs) was effective in enhancing the bioavailability of curcumin and improving the controlled drug release at the site of inflammation. Furthermore, in vitro and in vivo evaluation validate the capacity of HA@Cur/Cu to inhibit bacterial growth and remove excess ROS and inflammatory mediators, thereby significantly promoting the healing of diabetic wound in mice. These compelling findings strongly demonstrate the enormous promise of this multifunctional nanomedicine for the treatment of diabetic wound.

High-Performance Gel-Free and Label-Free Size Fractionation of Extracellular Vesicles with Two-Dimensional Electrophoresis in a Microfluidic Artificial Sieve.

Extracellular vesicles (EVs) are cell-derived particles that exhibit diverse sizes, molecular contents, and clinical implications for various diseases depending on their specific subpopulations. However, fractionation of EV subpopulations with high resolution, efficiency, purity, and yield remains an elusive goal due to their diminutive sizes. In this study, we introduce a novel strategy that effectively separates EV subpopulations in a gel-free and label-free manner, using two-dimensional (2D) electrophoresis in a microfluidic artificial sieve. The microfabricated artificial sieve consists of periodically arranged micro-slit-well structures in a 2D array and generates an anisotropic electric field pattern to size fractionate EVs into discrete streams and steer the subpopulations into designated outlets for collection within a minute. Along with fractionating EV subpopulations, contaminants such as free proteins and short nucleic acids can be simultaneously directed to waste outlets, thus accomplishing both size fractionation and purification of EVs with high performance. Our platform offers a simple, rapid, and versatile solution for EV subpopulation isolation, which can potentially facilitate the discovery of biomarkers for specific EV subtypes and the development of EV-based therapeutics.

Correction: Sub-nL thin-film differential scanning calorimetry chip for rapid thermal analysis of liquid samples.

Correction for 'Sub-nL thin-film differential scanning calorimetry chip for rapid thermal analysis of liquid samples' by Sheng Ni et al., Lab Chip, 2023, 23, 1926-1934, https://doi.org/10.1039/D2LC01094A.

Metronidazole-loaded polydopamine nanomedicine with antioxidant and antibacterial bioactivity for periodontitis.

Aim: This study focused on treating periodontitis with bacterial infection and local over accumulation of reactive oxygen species. Materials & methods: Polydopamine nanoparticles (PDA NPs) were exploited as efficient carriers for encapsulated metronidazole (MNZ). The therapeutic efficacy and biocompatibility of PDA@MNZ NPs were investigated through both in vitro and in vivo studies. Results: The nanodrug PDA@MNZ NPs were successfully fabricated, with well-defined physicochemical characteristics. In vitro, the PDA@MNZ NPs effectively eliminated intracellular reactive oxygen species and inhibited the growth of Porphyromonas gingivalis. Moreover, the PDA@MNZ NPs exhibited synergistic therapy for periodontitisin in vivo. Conclusion: PDA@MNZ NPs were confirmed with exceptional antimicrobial and antioxidant functions, offering a promising avenue for synergistic therapy in periodontitis.

Accelerated Electrophoretic Focusing and Purification of DNA Based on Synchronous Coefficient of Drag Alteration.

Synchronous coefficient of drag alteration refers to a multidimensional transport mechanism where a net drift of molecules is achieved under a zero-time-average alternating motive force by perturbing their drag coefficient synchronously with the applied force. An electrophoretic form of the method is often applied to focus and purify nucleic acids in a gel under rotating electric fields. However, this method requires lengthy operation due to the use of limited field strengths. Here, using DNA as target molecules, we demonstrate that the operation time can be reduced from hours to minutes by replacing polymer gel with a microfabricated artificial sieve. We also describe an electrophoretic protocol that facilitates the collection of purified DNA from the sieve, which is shown to yield amplifiable DNA from crude samples including the lysates of cultured cells and whole blood. The sieve can be further equipped with nucleic acid amplification and detection functions for a point-of-care diagnostic application.

Biomimetic nanoparticles to enhance the reverse cholesterol transport for selectively inhibiting development into foam cell in atherosclerosis.

A disorder of cholesterol homeostasis is one of the main initiating factors in the progression of atherosclerosis (AS). Metabolism and removal of excess cholesterol facilitates the prevention of foam cell formation. However, the failure of treatment with drugs (e.g. methotrexate, MTX) to effectively regulate progression of disease may be related to the limited drug bioavailability and rapid clearance by immune system. Thus, based on the inflammatory lesion "recruitment" properties of macrophages, MTX nanoparticles (MTX NPs) camouflaged with macrophage membranes (MM@MTX NPs) were constructed for the target to AS plaques. MM@MTX NPs exhibited a uniform hydrodynamic size around ~ 360 nm and controlled drug release properties (~ 72% at 12 h). After the macrophage membranes (MM) functionalized "homing" target delivery to AS plaques, MM@MTX NPs improved the solubility of cholesterol by the functionalized ß-cyclodextrin (ß-CD) component and significantly elevate cholesterol efflux by the loaded MTX mediated the increased expression levels of ABCA1, SR-B1, CYP27A1, resulting in efficiently inhibiting the formation of foam cells. Furthermore, MM@MTX NPs could significantly reduce the area of plaque, aortic plaque and cholesterol crystals deposition in ApoE-/- mice and exhibited biocompatibility. It is suggested that MM@MTX NPs were a safe and efficient therapeutic platform for AS.

Continuous-flow label-free size fractionation of extracellular vesicles through electrothermal fluid rolls and dielectrophoresis synergistically integrated in a microfluidic device.

Extracellular vesicles (EVs) are cell-derived bioparticles that play significant roles in various biological processes including cell-to-cell communication and intercellular delivery. Additionally, they hold great potential as liquid biopsy biomarkers for pre-diagnostic applications. However, the isolation of EV subpopulations, especially exosomes from a biological fluid remains a challenge due to their submicron range. Here, we demonstrate continuous-flow label-free size fractionation of EVs for the first time through a synergistic combination of electrothermal fluid rolls and dielectrophoresis in a microfluidic device. The device features three dimensional microelectrodes with unique sidewall contours that give rise to effective electrothermal fluid rolls in cooperation with dielectrophoretic forces for the electrokinetic manipulation and size separation of submicron particles. We first validate the device functionality by separating submicron polystyrene particles from binary mixtures with a cut-off size of ∼200 nm and then isolate intact exosomes from cell culture medium or blood serum with a high recovery rate and purity (∼80%). The device operation in a high-conductivity medium renders the method ideal for the purification of target bioparticles directly from physiological fluids, and may offer a robust and versatile platform for EV related diagnostic applications.

Sub-nL thin-film differential scanning calorimetry chip for rapid thermal analysis of liquid samples.

Differential scanning calorimetry (DSC) is a popular thermal analysis technique. The miniaturization of DSC on chip as thin-film DSC (tfDSC) has been pioneered for the analysis of ultrathin polymer films at temperature scan rates and sensitivities far superior to those attainable with DSC instruments. The adoption of tfDSC chips for the analysis of liquid samples is, however, confronted with various issues including sample evaporation due to the lack of sealed enclosures. Although the subsequent integration of enclosures has been demonstrated in various designs, rarely did those designs exceed the scan rates of DSC instruments mainly because of their bulky features and requirement for exterior heating. Here, we present a tfDSC chip featuring sub-nL thin-film enclosures integrated with resistance temperature detectors (RTDs) and heaters. The chip attains an unprecedented sensitivity of 11 V W-1 and a rapid time constant of 600 ms owing to its low-addenda design and residual heat conduction (∼6 µW K-1). We present results on the phase transition of common liquid crystals which we leverage to calibrate the RTDs and characterize the thermal lag with scan rates up to 900 °C min-1. We then present results on the heat denaturation of lysozyme at various pH values, concentrations, and scan rates. The chip can provide excess heat capacity peaks and enthalpy change steps without much alteration induced by the thermal lag at elevated scan rates up to 100 °C min-1, which is an order of magnitude faster than those of many chip counterparts.

Comparative Transcriptomics and Metabolites Analysis of Two Closely Related Euphorbia Species Reveal Environmental Adaptation Mechanism and Active Ingredients Difference.

Roots of Euphorbia fischeriana and Euphorbia ebracteolata are recorded as the source plant of traditional Chinese medicine "Langdu," containing active ingredients with anticancer and anti-AIDS activity. However, the two species have specific patterns in the graphic distribution. Compared with E. ehracteolata, E. fischeriana distributes in higher latitude and lower temperature areas and might have experienced cold stress adaptation. To reveal the molecular mechanism of environmental adaptation, RNA-seq was performed toward the roots, stems, and leaves of E. fischeriana and E. ehracteolata. A total of 6,830 pairs of putative orthologs between the two species were identified. Estimations of non-synonymous or synonymous substitution rate ratios for these orthologs indicated that 533 of the pairs may be under positive selection (Ka/Ks > 0.5). Functional enrichment analysis revealed that significant proportions of the orthologs were in the TCA cycle, fructose and mannose metabolism, starch and sucrose metabolism, fatty acid biosynthesis, and terpenoid biosynthesis providing insights into how the two closely related Euphorbia species adapted differentially to extreme environments. Consistent with the transcriptome, a higher content of soluble sugars and proline was obtained in E. fischeriana, reflecting the adaptation of plants to different environments. Additionally, 5 primary or secondary metabolites were screened as the biomarkers to distinguish the two species. Determination of 4 diterpenoids was established and performed, showing jolkinolide B as a representative component in E. fischeriana, whereas ingenol endemic to E. ebracteolate. To better study population genetics, EST-SSR markers were generated and tested in 9 species of Euphorbia. A total of 33 of the 68 pairs were screened out for producing clear fragments in at least four species, which will furthermore facilitate the studies on the genetic improvement and phylogenetics of this rapidly adapting taxon. In this study, transcriptome and metabolome analyses revealed the evolution of genes related to cold stress tolerance, biosynthesis of TCA cycle, soluble sugars, fatty acids, and amino acids, consistent with the molecular strategy that genotypes adapting to environment. The key active ingredients of the two species were quantitatively analyzed to reveal the difference in pharmacodynamic substance basis and molecular mechanism, providing insights into rational crude drug use.

Ordered surface crack patterns in situ formed under confinement on fluidic microchannel boundaries in polydimethylsiloxane.

We present ordered surface crack patterns discovered in microfluidic channels/chambers in polydimethylsiloxane (PDMS). The cracks are formed in situ under confinement due to compression applied following an oxygen plasma step in a soft lithography process. The crack patterns are noticeable only after fluorescent labeling and vary with fluidic layout as well as material compliance.

Mercury wet depositions study at suburban, agriculture and traffic sampling sites.

This study aimed to measure and discuss the relationship of ambient air precipitations with respect to mercury wet depositions at suburban, agriculture and traffic three characteristic sampling sites during the year of 2019. In addition, the mercury volume weighted mean concentrations (VWM) at three characteristic sampling sites were also calculated. Finally, the ambient mercury wet depositions data obtained in this study to various world sampling sites were also compared and discussed in this study. The results indicated that the average mercury wet depositions for suburban, agriculture and traffic areas were 0.62, 0.55 and 2.32 ng/m2 min, respectively. And the average mercury VWM values were 0.9, 0.72 and 1.85 ng/m2 min for suburban, agriculture and traffic sites, respectively. In addition, the highest VWM and wet depositions for mercury both occurred in March at traffic and suburban areas. And the mercury wet depositions displayed a declined trend when the month was moved from March to July at both traffic and suburban sampling sites. In addition, the relationship between wet depositions and precipitations was low to moderate correlated in traffic area, while the relationship between wet depositions and precipitations was insignificant at both suburban and agriculture areas. Moreover, the average highest mercury wet deposition occurred in Nepal when compared to the other world sites. In addition, the average value of mercury wet depositions in Nepal was about 17.23 times to that of data obtained in this study during the period of 2007-2019. Finally, the average highest VWM (ng/L) occurred in the China. In addition, the average value mercury VWM in China was about 14.82 times to that of data obtained in this study during the period of 2007-2019.

ADORA1 Promotes Hepatocellular Carcinoma Progression via PI3K/AKT Pathway.

OBJECTIVE: Hepatocellular carcinoma (HCC) is a common malignancy worldwide. Although the contradictory role of ADORA1 has been explored in certain types of cancers, its clinical significance and function in hepatocellular carcinoma cells are largely unknown. MATERIALS AND METHODS: The level of ADORA1 in HCC tissues and cells was evaluated by RT-PCR. The function of ADORA1 overexpression on HCC cell proliferation and invasion was assessed by MTS, transwell analysis, and colony formation assay. In addition, a mouse subcutaneous xenograft model was used to study in vivo effects. The efficacy of knockdown of ADORA1 sensitizes to chemotherapy was assessed by staining with Annexin V/propidium iodide followed with flow cytometry and nuclei fragmentation. RESULTS: In this study, ADORA1 was identified to be up-regulated in HCC tissues compared with adjacent normal tissue. High ADORA1 mRNA expression predicted poor survival in hepatocellular carcinoma patients. Ectopic expression of ADORA1 increased hepatocellular carcinoma cell proliferation and invasion. ADORA1 knockdown inhibited HCC cell growth and sensitized to chemotherapy. Furthermore, ADORA1 activated PI3K/AKT oncogenic signaling pathways. Treatment with PI3K inhibitor LY294002 blocked the effects of ADORA1 on tumor growth in either ADORA1-overexpressing or -deficiency cells. Finally, overexpression of ADORA1 stimulates HCC tumor growth in vivo. Treatment of ADORA1 antagonist oppositely suppressed HCC xenograft tumor growth. CONCLUSION: ADORA1 serves as an important oncoprotein and a promoter of cell proliferation through PI3K/AKT signaling pathway in hepatocellular carcinoma.

The vision of point-of-care PCR tests for the COVID-19 pandemic and beyond.

Infectious diseases, such as the most recent case of coronavirus disease 2019, have brought the prospect of point-of-care (POC) diagnostic tests into the spotlight. A rapid, accurate, low-cost, and easy-to-use test in the field could stop epidemics before they develop into full-blown pandemics. Unfortunately, despite all the advances, it still does not exist. Here, we critically review the limited number of prototypes demonstrated to date that is based on a polymerase chain reaction (PCR) and has come close to fulfill this vision. We summarize the requirements for the POC-PCR tests and then go on to discuss the PCR product-detection methods, the integration of their functional components, the potential applications, and other practical issues related to the implementation of lab-on-a-chip technologies. We conclude our review with a discussion of the latest findings on nucleic acid-based diagnosis.

Transcriptome and metabolome analysis reveals regulatory networks and key genes controlling barley malting quality in responses to drought stress.

Malting quality will be greatly deteriorated when barley plants suffer from post-anthesis drought stress, however there is a marked difference among barley genotypes in the responses of malting quality to drought stress, and the molecular mechanisms underlying the genotypic difference remain unclear. We made transcriptome and metabolome analysis on the developing grains of two barley genotypes differing in the responses to drought stress. Post-anthesis drought treatments led to decreased grain weight and ß-glucan content, increased grain protein content and ß-amylase activity. Drought stress enhanced H2O2 and heat-shock protein accumulation in the two barley genotypes, with the drought-tolerant genotype showing higher capacity of scavenging H2O2 and reducing misfolded protein accumulation than the drought-susceptible genotype. Moreover, the drought-tolerant genotype was more efficient in redistributing assimilates stored in the vegetative tissues into the developing grains. After re-watering to relieve drought stress, the drought-tolerant genotype can further modify auxin transport and ethylene signaling, enhancing redistribution of assimilates into grains. Transcriptome comparisons and weighted correlation network analysis (WGCNA) identified some key genes regulating the responses of malting quality traits to drought stress, such as RLK-LRR, ß-glucosidase and HSP . In conclusion, less change of main malting quality traits in the drought-tolerant genotype under post-anthesis drought stress is attributed to its higher capacity of alleviating the stress injury through scavenging ROS and redistributing the metabolites stored in the vegetative organs into the developing grains.

Rapid Characterization of Biomolecules' Thermal Stability in a Segmented Flow-Through Optofluidic Microsystem.

Optofluidic devices combining optics and microfluidics have recently attracted attention for biomolecular analysis due to their high detection sensitivity. Here, we show a silicon chip with tubular microchannels buried inside the substrate featuring temperature gradient (∇T) along the microchannel. We set up an optical fluorescence system consisting of a power-modulated laser light source of 470 nm coupled to the microchannel serving as a light guide via optical fiber. Fluorescence was detected on the other side of the microchannel using a photomultiplier tube connected to an optical fiber via a fluorescein isothiocyanate filter. The PMT output was connected to a lock-in amplifier for signal processing. We performed a melting curve analysis of a short dsDNA - SYBR Green I complex with a known melting temperature (TM) in a flow-through configuration without gradient to verify the functionality of the proposed detection system. We then used the segmented flow configuration and measured the fluorescence amplitude of a droplet exposed to ∇T of ≈ 2.31 °C mm-1, determining the heat transfer time as ≈ 554 ms. The proposed platform can be used as a fast and cost-effective system for performing either MCA of dsDNAs or for measuring protein unfolding for drug-screening applications.

Human Neutrophil Elastase Activated Fluorescent Probe for Pulmonary Diseases Based on Fluorescence Resonance Energy Transfer Using CdSe/ZnS Quantum Dots.

There is an increasing demand for effective noninvasive diagnosis against common pulmonary diseases, which are rising sharply due to the serious air pollution. Human neutrophil elastase (HNE), a typical protease highly involved in pulmonary inflammatory diseases and lung cancer, is a potential predictor for disease progression. Currently, few of the HNE-targeting probes are applicable in vivo due to the limitation in sensitivity and biocompatibility. Herein, we reported the achievement of in vitro detection and in vivo imaging of HNE by incorporating the HNE-specific peptide substrate, quantum dots (QDs), and organic dyes into the fluorescence resonance energy transfer (FRET) system. The refined nanoprobe, termed QDP, could specifically measure the HNE with excellent sensitivity of 7.15 pM in aqueous solution and successfully image the endogenous and exogenous HNE in living cells. In addition, this nanoprobe enabled HNE imaging in mouse models of lung cancer and acute lung injury, and the HNE activity at high temporal and spatial resolution was continuously monitored. Most importantly, QDP successfully discriminated the serums of patients with lung diseases from those of the healthy controls based on the HNE activity determination. Overall, this study demonstrates the advantages of a FRET-system-based nanoprobe in imaging performance and provides an applicable tool for in vivo HNE detection and pulmonary disease diagnosis.

Detection of Plasma EGFR Mutations in NSCLC Patients with a Validated ddPCR Lung cfDNA Assay.

Purpose: The clinical utility of cell-free DNA (cfDNA) to assess EGFR mutations is increasing. However, there are limited studies determining their clinical validity and utility. The value of cfDNA assays in cancer management remains controversial. Methods: In this study, we first evaluated the analytical performance of the ddPCR Lung cfDNA Assay. We next analyzed the concordance of the results with tissue amplification refractory mutation system PCR (ARMS-PCR) and plasma next-generation sequencing (NGS) genotyping. Finally, we assessed its clinical utility by exploring the association of cfDNA EGFR mutations with metastatic sites and the efficacy of EGFR-TKIs treatment. Results: The ddPCR Lung cfDNA Assay demonstrated a limit of blank of 1 droplet per reaction, an analytical specificity of 100%, and detection limit of 0.05%, 0.05%, and 0.1% for E746_A750del, L858R, and T790M, respectively. With tissue ARMS-PCR as a standard for comparison, the clinical sensitivity and specificity of ddPCR were 62.5% (15/24) and 100% (82/82) for E746_A750del, and 75.0% (15/20) and 94.2% (81/86) for L858R, respectively. The ddPCR showed high concordance with NGS in determining cfDNA EGFR mutations. Patients with bone and/or brain metastasis showed a higher detection rate and mutant abundance of cfDNA EGFR mutations compared to those with other sites of metastasis. Moreover, EGFR-TKIs treatment was effective in patients with sensitive EGFR mutations in either plasma cfDNA or tumor tissue-derived DNA. Conclusions: We validated in this study that the ddPCR Lung cfDNA Assay is reliable for detection of EGFR mutations in lung cancers, in terms of analytical performance, clinical validity and utility.

Primary Ewing's sarcoma of the vertebral body: A case report.

RATIONAL: The occurrence of Ewing's sarcoma in the vertebral body of elderly women is extremely rare, and the case of Ewing's sarcoma in the spine with secondary surgical repair after wrong diagnosis and treatment has not been reported. We report a case involving primary Ewing's sarcoma of the vertebral body in an elderly female. Owing to its rarity and controversial issues, we report a case report to discuss its clinical features, treatments, radiological, and histological characteristics. PATIENT CONCERNS: The elderly female patient came to see us with the manifestation of total paralysis of both lower limbs. The patient with a vertebral compression fracture as the primary manifestation was misdiagnosed in another hospital. The patient underwent inappropriate surgical treatment and was transferred to our hospital for diagnosis and second-stage surgery. DIAGNOSES: The postoperative pathological examination and immunohistochemical examination in our hospital confirmed: Ewing's sarcoma; Surgical history at other hospitals suggests: after Bone cement injection. INTERVENTIONS: The patient underwent a T6 and T8 laminectomy and T5/6-T9 pedicle screw fixation. OUTCOMES: Reexamination 1 month after the surgery showed that the tumor had been partially resected, the spinal cord compression was relieved, the tumor did not grow further, and the patient's lower limb physical ability, tactile sense, algesia and temperature sense recovered slightly. LESSONS: For patients with ewing's tumor in the spinal canal with symptoms of spinal cord compression, even if the patients with poor results after a unadvisable operation, it is still necessary to be actively in spinal cord compression by surgery. The differential diagnosis of Ewing's sarcoma and compression fractures is very important. For patients with vertebral tumors, special attention should be taken during vertebroplasty for bone cement leakage caused by excessive bone cement injection and increased local pressure. And some experience with imaging and laboratory findings.

Learning to Predict Drug Target Interaction From Missing Not at Random Labels.

The prediction of Drug-Target Interaction (DTI) is an important research direction in bioinformatics as it greatly shortens the development cycle of new drugs. State-of-the-art computational methods for DTI prediction adopt a binary classification framework. The supervision is incomplete, i.e. only a small amount of DTIs are known and treated as positive instances, while the rest are unknown and treated as negative. Two severe problems occur in such a framework: (1) the number of negative samples is overwhelming and (2) a negative label cannot rule out the possibility of a positive drug-target interaction. In this paper, we address the problem of learning from incomplete labels in DTI prediction. The key assumption here is that labels are missing not at random. For example, negative DTI labels are more likely to be missing because biomedical researchers prioritize to study DTIs that are more likely to be positive. We introduce a novel probabilistic model, factorization with non-random missing labels (FNML). It models the generative process for the DTI labels (i.e. the labels are positive or negative) and responses (i.e. the labels are observed or missing). In particular, the probability of observing or missing a label is associated with the sign of the label. In order to further reduce prediction variance and improve prediction accuracy on highly imbalanced DTI datasets, we present FNML-EN, an ensemble scheme which is designed specifically for FNML model. We conduct comprehensive experiments on the latest DTI database, demonstrating the superior and robust performance of the proposed models.

Jones pupil metrology of lithographic projection lens and its optimal configuration in the presence of error sources.

Mueller matrix imaging polarimeter (MMIP) can be used to measure the polarization aberration (PA) of a lithographic projector in the form of the Mueller pupil, while the Jones pupil is required for lithographic imaging simulations, projection lens design and PA evaluation. In this paper, a Jones pupil measurement method of lithographic projection lens is proposed. The measurement device of the method is the same as an MMIP, but a new polarimetric measurement equation is derived to solve the Jones pupil directly from the Kronecker product of the Jones matrix and the measured intensities. Two new polarimeter configurations with the minimum condition number are designed to further improve the accuracy in the presence of error sources. The performance of the method is evaluated by measurement errors of a typical Jones pupil in the presence of error sources. Comparisons between the proposed method and the conventional method, in which the Jones pupil is converted from the Mueller pupil measured by MMIP, are given. The results validate that the measurement accuracy of the Jones pupil is significantly improved without increasing the complexity of existing measurement systems.