Particle separation technology plays an important role in a wide range of applications as a critical sample preprocessing step for analysis. In this work, we proposed and fabricated a multilayer ...lateral-flow particle filtration and separation device based on polydimethylsiloxane molding and transfer bonding techniques. Particle separation capability was demonstrated by 4.5-um polystyrene bead filtration and cancer cell (SK-BR-3) retrieving. This device exhibits higher throughput compared with most active particle separation methods and is less vulnerable to membrane clogging problem. This novel multilayer particle filtration and separation device is expected to find applications in biomedical, environmental and microanalysis fields.
For ultraviolet nanoimprint lithography (UV-NIL), the resist volume shrinkage during curing not only influences the pattern fidelity but also induces defects in the demolding process due to strong ...adhesion. To address this issue, a novel nanoimprint resist was formulated and characterized in this work. The new resist formulation contains 3,9-diethyl- 3,9-bis(allyloxymethyl)-1,5,7,11- tetraoxastetraoxaspiro 5 undecane (DB-TOSU), which is a liquid spiroorthocarbonate monomer that undergoes volume expansion upon acid-catalyzed polymerization. By mixing DB-TOSU with conventional volume-shrinking epoxy monomers at various weight ratios, the formulated resists had much reduced or even zero volume shrinkage. The resist volume shrinkage, elastic modulus, and demolding force decreased with increasing DB-TOSU weight ratio in resist formulation. When DB-TOSU reached 50 wt%, the demolding force was reduced by 69% with an adequate elastic modulus (75 MPa) and low shrinkage (1.86%). This novel resist formulation has the potential to allow high-fidelity pattern replication and reduce demolding defects in UV-NIL.
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•The liquid volume-expanding monomer DB-TOSU was successfully synthesized.•New UV nanoimprint resists were formulated with low volume shrinkage.•The new resists significantly reduce demolding forces in nanoimprint.
•A risk prediction for IDH in HD-patients can be an important tool for clinical work.•LightGBM model plays as an interpretable and best-performing model for the task.•IDH-A and IDH-B model can ...usefully complement each other for risk prediction.
Intradialytic hypotension (IDH) is closely associated with adverse clinical outcomes in HD-patients. An IDH predictor model is important for IDH risk screening and clinical decision-making. In this study, we used Machine learning (ML) to develop IDH model for risk prediction in HD patients.
62,227 dialysis sessions were randomly partitioned into training data (70%), test data (20%), and validation data (10%). IDH-A model based on twenty-seven variables was constructed for risk prediction for the next HD treatment. IDH-B model based on ten variables from 64,870 dialysis sessions was developed for risk assessment before each HD treatment. Light Gradient Boosting Machine (LightGBM), Linear Discriminant Analysis, support vector machines, XGBoost, TabNet, and multilayer perceptron were used to develop the predictor model.
In IDH-A model, we identified the LightGBM method as the best-performing and interpretable model with C- statistics of 0.82 in Fall30Nadir90 definitions, which was higher than those obtained using the other models (P<0.01). In other IDH standards of Nadir90, Nadir100, Fall20, Fall30, and Fall20Nadir90, the LightGBM method had a performance with C- statistics ranged 0.77 to 0.89. As a complementary application, the LightGBM model in IDH-B model achieved C- statistics of 0.68 in Fall30Nadir90 definitions and 0.69 to 0.78 in the other five IDH standards, which were also higher than the other methods, respectively.
Use ML, we identified the LightGBM method as the good-performing and interpretable model. We identified the top variables as the high-risk factors for IDH incident in HD-patient. IDH-A and IDH-B model can usefully complement each other for risk prediction and further facilitate timely intervention through applied into different clinical setting.
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Gemcitabine is the first-line chemotherapy drug for cholangiocarcinoma (CCA), but acquired resistance has been frequently observed in CCA patients. To search for potential long noncoding RNAs ...(lncRNAs) involved in gemcitabine resistance, two gemcitabine resistant CCA cell lines were established and dysregulated lncRNAs were identified by lncRNA microarray. Long intergenic non-protein coding RNA 665 (LINC00665) were found to rank the top 10 upregulated lncRNAs in our study, and high LINC00665 expression was closely associated with poor prognosis and chemoresistance of CCA patients. Silencing LINC00665 in gemcitabine resistant CCA cells impaired gemcitabine tolerance, while enforced LINC00665 expression increased gemcitabine resistance of sensitive CCA cells. The gemcitabine resistant CCA cells showed increased EMT and stemness properties, and silencing LINC00665 suppressed sphere formation, migration, invasion and expression of EMT and stemness markers. In addition, Wnt/β-Catenin signaling was activated in gemcitabine resistant CCA cells, but LINC00665 knockdown suppressed Wnt/β-Catenin activation. B-cell CLL/lymphoma 9-like (BCL9L), the nucleus transcriptional regulators of Wnt/β-Catenin signaling, plays a key role in the nucleus translocation of β-Catenin and promotes β-Catenin-dependent transcription. In our study, we found that LINC00665 regulated BCL9L expression by acting as a molecular sponge for miR-424-5p. Moreover, silencing BCL9L or miR-424-5p overexpression suppressed gemcitabine resistance, EMT, stemness and Wnt/β-Catenin activation in resistant CCA cells. In conclusion, our results disclosed the important role of LINC00665 in gemcitabine resistance of CCA cells, and provided a new biomarker or therapeutic target for CCA treament.
The therapeutic efficacy of stem cells transplanted into an ischaemic brain depends primarily on the responses of the neurovascular unit. Here, we report the development and applicability of a ...functional neurovascular unit on a microfluidic chip as a microphysiological model of ischaemic stroke that recapitulates the function of the blood-brain barrier as well as interactions between therapeutic stem cells and host cells (human brain microvascular endothelial cells, pericytes, astrocytes, microglia and neurons). We used the model to track the infiltration of a number of candidate stem cells and to characterize the expression levels of genes associated with post-stroke pathologies. We observed that each type of stem cell showed unique neurorestorative effects, primarily by supporting endogenous recovery rather than through direct cell replacement, and that the recovery of synaptic activities is correlated with the recovery of the structural and functional integrity of the neurovascular unit rather than with the regeneration of neurons.
•HOTAIR expression was increased in cholangiocarcinoma.•HOTAIR deletion inhibited cholangiocarcinoma progression.•HOTAIR upregulated HMGB1 expression by sponging miR-204-5p.•HOTAIR promoted ...cholangiocarcinoma progression via miR-204-5p/HMGB1 axis.
Cholangiocarcinoma (CCA) is a malignant tumor in the world. LncRNA HOX transcript antisense intergenic RNA (HOTAIR) was identified as a crucial regulator in various cancers including CCA. This study aimed to unravel the functions of HOTAIR and its biological mechanism in CCA, hinting for the new therapeutic targets in CCA.
The levels of HOTAIR, miR-204-5p and HMGB1 in CCA tissues and cell lines (HuB28 and HuCCT1) were measured by quantitative real-time polymerase chain reaction (qRT-PCR). Western blot was conducted to detect the protein levels of LC3-I, LC3-II, Beclin-1 and HMGB1. The relationships among HOTAIR, miR-204-5p and HMGB1 were examined by dual-luciferase reporter assay, RNA immunoprecipitation (RIP) assay and RNA pull down assay. Cell proliferation ability and apoptosis rate were assessed by CCK8 assay and flow cytometry, respectively. in vivo experiment was conducted to examine the bio-functions of HOTAIR in nude mice.
HOTAIR and HMGB1 were over-expressed, while miR-204-5p was lowly expressed in CCA tissues and cells. The dual-luciferase reporter assay indicated that miR-204-5p was a target of HOTAIR, and HMGB1 was a target of miR-204-5p. The restoration experiments showed that HOTAIR repressed cell apoptosis, autophagy and promoted cell proliferation via miR-204-5p/HMGB1 axis. Additionally, HOTAIR silencing retarded the xenograft tumor growth by up-regulation of miR-204-5p and down-regulation of HMGB1.
These data unraveled that lncRNA HOTAIR regulated HMGB1 to suppress cell apoptosis, autophagy and induce cell proliferation by sponging miR-204-5p in CCA. Thus, this new regulatory pathway may provide new therapeutic targets for CCA.
Erythromycin fermentation residue (EFR) is a solid waste generated from the fermentation process of erythromycin A production. Some byproducts are produced during the fermentation process of ...erythromycin A production, and erythromycin A can also undergo hydrolysis and biodegradation reactions in the environment with the formation of transformation products. Herein, an accurate analytical method was established and validated to quantify erythromycin A, two byproducts and five hydrolysis or biodegradation products, in solid or semi-solid media of waste EFR and the amended soil. The method mainly included ultrasonic solvent extraction, solid phase extraction, and ultra-performance liquid chromatography-tandem mass spectrometry quantification. All analytes could be effectively extracted in a single process, and the recoveries ranged from 76% to 122% for different matrices. Low matrix effects and excellent precision were achieved by optimizing the mass spectrometry parameters, extraction solution, number of extractions and eluent. This method was applied to evaluate the residual analytes in EFR, treated EFR after industrial-scale hydrothermal treatment, and the subsequent soil application. Seven analytes were detected in the EFR, while six were found in the treated EFR and amended soils. The concentration of erythromycin A in EFR was 1,629 ± 100 mg/kg·TS, and the removal efficiency of hydrothermal treatment (180 °C, 60 min) was about 99.6%. Three hydrolysis products were the main residuals in treated EFR, with anhydroerythromycin A showing the highest concentration. The concentrations of the analytes in soil ranged from 2.17 ± 1.04 to 92.33 ± 20.70 μg/kg·TS, and anhydroerythromycin A contributed 65%–77% of the total concentration. Erythromycin B, a byproduct, was still detected in soil. This work provides an accurate analytical method which would be useful to evaluate the potential risk of byproducts and transformation products of erythromycin A in environment.
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•Erythromycin A and seven transformation products were quantified by UPLC-MS/MS.•Solid phase extraction was optimized to extract analytes from complex matrices.•Seven analytes were detected in erythromycin fermentation residue (EFR), while six were in treated EFR and amended soil.•Three hydrolysis products of ERY-A were the main transformation products in EFR and amended soil.
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•Oxytetracycline hydrolysis products (TPs) after enhanced hydrolysis were investigated.•Three known and 15 unknown TPs were identified by target and non-target analysis.•Eight yet ...unknown TPs were detected even after biological treatment in field studies.•Two dominant TPs (M390s) were isolated, purified, identified and quantified.•M390s could contribute 4.5% − 7.8% of COD in biological effluent.Graphical abstract.
With the increasing demand for stringent discharge regulations, the efficient removal of non-biodegradable organic (hard COD) from the biological effluent of pharmaceutical wastewater is becoming increasingly important. Enhanced hydrolysis pretreatment has been used to remove antibiotic potency in antibiotic production wastewater before biological treatment, while the resulting transformation products (TPs) and their contribution to hard COD are largely unknown. Herein, we combine target and non-target analysis aiming to find the major oxytetracycline (OTC) TPs after enhanced hydrolysis pretreatment and to quantify their contribution to the hard COD in biological effluents. Three literature-known and 15 yet unknown TPs, most of which could not be biodegraded, were identified during lab-scale hydrolysis of OTC under neutral conditions. Nuclear magnetic resonance and combined density functional theory were used to determine and infer the molecular structures and possible hydrolysis pathways of them. Furthermore, two stable and dominant TPs, M390s (M390-1 and M390-2), were separated, purified, and structurally identified, which could account for 21.0% and 14.9% − 20.4% (molar ratio) of the initial OTC in the lab-scale and field studies, respectively. More importantly, M390s account for 4.5–––7.8% of the hard COD in biological effluents. The results indicate that the contribution of antibiotic TPs to hard COD in wastewater could not be ignored, and provide more knowledge regarding the efficient treatment of antibiotic production wastewater.
•Five Ery-A-derived compounds are proved to contribute to the antibacterial activity.•Ery-A-related compounds lead to near collapse of the first-stage anaerobic tank.•High abundances of MLS ...resistance genes are generated in the treatment system.•The discharge of Ery-A-related compounds and resistance genes is nonnegligible.
Antibiotic production wastewater from pharmaceutical manufacturing is a significant source of antibiotic and resistance gene pollution in the environment. Given that Erythromycin A (Ery-A) is a widely used antibiotic in both human clinical and livestock breeding, it is imperative to ascertain its presence, along with related compounds, in the biological treatment processes of production wastewater. In this study, the occurrence and behavior of Ery-A, its production byproducts, transformation products, and resistance genes were first systematically investigated in a full-scale anaerobic-aerobic system for treating Ery-A production wastewater. Simultaneously, residual antibacterial activity in wastewater and sludge was evaluated throughout the wastewater treatment process. Ery-A contributes only 24.2 – 36.0% to the antibacterial activities. Ery-A-derived compounds including production byproducts (erythromycin B and erythromycin C) and transformation products (anhydro erythromycin A, N-demethyl-erythromycin A, and erythromycin A enol ether), are determined to contribute to the antibacterial activities of the wastewater treatment system. High concentrations of antibiotics with antibacterial activity (up to 1,258.9 mg/kg·TS for erythromycin A enol ether) adsorbed in the sludge result in near collapse of the first-stage anaerobic sludge system. Sludge biodegradation in second-stage anaerobic and anoxic-aerobic tanks is essential in removing Ery-A-related compounds from wastewater. The Ery-A-related compounds in the secondary effluent and excess sludge are determined to be 44.5 g/h and 1.5 g/h through the mass balance analysis, respectively. The discharge of MLS resistance genes from the secondary effluent and excess sludge is 1.0 × 1016 copies/h and 7.1 × 1015 copies/h, respectively. These findings highlight the significant concern over the release of Ery-A-related compounds and MLS resistance genes from the Ery-A production wastewater treatment system. As a result, it is crucial to implement strategies for the removal of Ery-A-related compounds from production wastewater before biological processes. This study is the first to report the occurrence and behavior of Ery-A-related compounds and resistance genes along the full-scale wastewater treatment processes. Additionally, it sheds light on the importance of byproducts and transformation products with antibacterial activity from Ery-A in the Ery-A production wastewater treatment system.
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