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Abstract Introduction Breast cancer ranks second as the most common malignancy globally, after lung cancer. Among the various subtypes of breast cancer, HER2 positive breast cancer (HER2 BC)poses a particularly challenging prognosis due to its heightened invasiveness and metastatic potential. The objective of this study was to construct a composite piezoelectric nanoparticle based on poly(vinylidene fluoride-trifluoroethylene) (P(VDF-TrFE)) for imaging and treatment of HER2 BC. Method By reshaping the crystal structure of P(VDF-TrFE) piezoelectric nanoparticles, improving hydrophilicity, and incorporating imaging capabilities, we developed piezoelectric composite nanoparticles (PGd@tNBs) that integrate imaging and therapeutic functions. The in vitro characterization encompassed the assessment of piezoelectric properties, hydrophilicity, imaging performance, and therapeutic efficacy of these particles. The targeting and therapeutic effectiveness of PGd@tNBs particles were further validated in the SK-BR3 cell line and subsequently confirmed in HER2-positive tumor-bearing mice. Results The nanoparticle demonstrated excellent biocompatibility and impressive multimodal imaging performance. Magnetic resonance imaging (MRI) observations revealed significant accumulation of PGd@tNBs particles in the HER2 positive tumor, exhibiting superior contrast-enhanced ultrasound performance compared to traditional ultrasound contrast agents, and small animal in vivo imaging showed that PGd@tNBs particles were primarily excreted through respiration and urinary metabolism. Piezoforce Microscopy characterization highlighted the outstanding piezoelectric properties of PGd@tNBs particles. Upon targeted binding to HER2-BC, ultrasound stimulation influenced the cell membrane potential, leading to reversible electroporation. This, in turn, affected the balance of calcium ions inside and outside the cells and the mitochondrial membrane potential. Following ingestion by cells, PGd@tNBs, when exposed to ultrasound, triggered the generation of reactive oxygen species (ROS), resulting in the consumption of glutathione and superoxide dismutase and achieving sonodynamic therapy. Notably, repeated ultrasound stimulation, post PGd@tNBs particles binding and entry into cells, increased ROS production and elevated the apoptosis rate by approximately 45%. Conclusion In conclusion, the PGd@tNBs particles developed exhibit outstanding imaging and therapeutic efficacy, holding potential for precise diagnosis and personalized treatment of HER2 BC.

Introduction: Patients with advanced primary liver cancer often lose the opportunity for surgery when they are found, and the treatment options are limited. Lenvatinib, as a multi-target tyrosine kinase inhibitor, has been used as the first-line treatment for advanced liver cancer. Immune checkpoint inhibitors, such as programmed cell death protein 1 inhibitors, have been successfully used in advanced or metastatic liver cancer. Case Presentation: We report a case of combined lenvatinib and the programmed cell death protein 1 inhibitor camrelizumab in the treatment of primary liver cancer, in which the rare complication of full-thickness gastric mucosa exfoliation occurred. To the best of our knowledge, this is the first report of the side effect of hemorrhagic exfoliative gastritis with the combination of lenvatinib and camrelizumab. Conclusion: Hemorrhagic exfoliative gastritis is an extremely rare clinical complication. Lenvatinib inhibits vascular proliferation and could cause gastrointestinal perforation, which is considered to be the main factor, but whether camrelizumab plays a role in it or only causes gastrointestinal reactions leading to nausea and vomiting, resulting in gastric mucosal exfoliation bleeding, remains to be further explored.

Abstract Purpose To construct a gadoxetic acid-enhanced MRI (EOB-MRI) -based multivariable model to predict Ki-67 expression levels in hepatocellular carcinoma (HCC) using LI-RADS v2018 imaging features. Methods A total of 121 patients with HCC who underwent EOB-MRI were enrolled in this study. The patients were divided into three groups according to Ki-67 cut-offs: Ki-67 ≥ 20% (n = 86) vs. Ki-67 0.05). Four models were constructed including model A (mosaic architecture and infiltrated appearance), model B (mosaic architecture and targetoid HBP), model C (infiltrated appearance and targetoid HBP), and model D (mosaic architecture, infiltrated appearance and targetoid HBP). The model D yielded better diagnostic performance than the model C (0.776 vs. 0.669, p = 0.002), but a comparable AUC than model A (0.776 vs. 0.781, p = 0.855) and model B (0.776 vs. 0.746, p = 0.076). Conclusions Mosaic architecture, infiltrated appearance and targetoid HBP were sensitive imaging features for predicting Ki-67 index ≥ 50% and EOB-MRI model based on LI-RADS v2018 features may be an effective imaging approach for the risk stratification of patients with HCC before surgery.

Circulating tumor cells (CTCs) have significant clinical value in early tumor detection, dynamic monitoring and immunotherapy. CTC detection stands out as a leading non-invasive approach for tumor diagnostics and therapeutics. However, the high heterogeneity of CTCs and the occurrence of epithelial-mesenchymal transition (EMT) during metastasis pose challenges to methods relying on EpCAM-positive enrichment. To address these limitations, a method based on negative enrichment of CTCs using specific leukocyte targets has been developed. In this study, aiming to overcome the low purity associated with immunomagnetic beads targeting solely the leukocyte common antigen CD45, we introduced CD66b-modified immunomagnetic beads. CD66b, a specific target for neutrophils with abundant residues, was chosen as a complementary approach. The process involved initial collection of nucleated cells from whole blood samples using density gradient centrifugation. Subsequently, magnetically labeled leukocytes were removed by magnetic field, enabling the capture of CTCs with higher sensitivity and purity while retaining their activity. Finally, we selected 20 clinical blood samples from patients with various cancers to validate the effectiveness of this strategy, providing a new generalized tool for the clinical detection of CTCs.

ABSTRACT Heat stress is one of the major stressors affecting dairy cow production, which causes great losses in animal husbandry production and economic development. This study combined 16S rDNA sequencing, metagenomic sequencing, and metabolomic analysis to investigate the effects of heat stress on the rumen microbiome and metabolism of cows. To achieve this, 10 cows each of growing heifers, heifers, and lactating cows were selected for sample collection in April and August (a total of 60 cows). Ruminal fluid was collected, filtered through gauze, and immediately transferred to liquid nitrogen prior to metagenomic, 16S rDNA sequencing, and metabolomic analyses. Heat stress increased the abundance of pathogenic bacteria, such as Treponema_2, Sphingobacterium, and Streptococcus, in cattle at all growth stages, with the structure and composition of rumen microbial communities being the most affected by heat stress in growing heifers and the least affected by heat stress in lactating cows. Heat stress led to a significant decrease in carbohydrate metabolic function in growing heifers and heifers, whereas microbial function was the most stable in lactating cows, suggesting that changes in microbial function are not only more conserved than microbial composition but are also affected by age. Heat stress led to significant enrichment of fatty acyl metabolites in growing heifers, heifers, and lactating cows, and the heat stress indicator species Bacillus, Lactococcus, Streptococcus, and Enterococcus were significantly associated with fatty acyl compounds in growing heifers and heifers. Therefore, heat stress alters the metabolic status of cows by modulating fatty acid biosynthesis, the PPAR signaling pathway, and arachidonic acid metabolism through the regulation of long-chain fatty acids and other biomarkers in cows at three growth stages. IMPORTANCE Heat stress is one of the main causes of economic losses in the dairy industry worldwide; however, the mechanisms associated with the metabolic and microbial changes in heat stress remain unclear. Here, we characterized both the changes in metabolites, rumen microbial communities, and their functional potential indices derived from rumen fluid and serum samples from cows at different growth stages and under different climates. This study highlights that the rumen microbe may be involved in the regulation of lipid metabolism by modulating the fatty acyl metabolites. Under heat stress, the changes in the metabolic status of growing heifers, heifers, and lactating cows were closely related to arachidonic acid metabolism, fatty acid biosynthesis, and energy metabolism. Moreover, this study provides new markers for further research to understand the effects of heat stress on the physiological metabolism of Holstein cows and the time-dependent changes associated with growth stages.

A new sol-gel method that employs cation exchange from an aqueous metal ion solution with H+ ions of granulated alginic acid was developed for synthesizing high-purity Y2O3 nanoparticles. In this study, the cation exchange kinetics of H+~Y3+ in aqueous solution were analyzed using on-line pH technology and off-line inductively coupled plasma-atomic emission spectrometry (ICP-AES) analysis. Pseudo 2nd-order models were utilized to evaluate the parameters of the kinetics, suggesting that the concentration of H+~Y3+ involved in the cation exchange reaction was 1:1.733. Further, a comprehensive understanding of the Y-ALG calcination process was developed using thermo-gravimetric analysis, along with results obtained from differential scanning calorimetry (TGA/DSC). A detailed analysis of the XRD Rietveld refinement plots revealed that the crystallite sizes of Y2O3 nanoparticles were about 4 nm (500 °C) and 15 nm (800 °C), respectively. Differential pulse voltammetry (DPV) was employed to investigate the electrochemical oxidation of catechol. The oxidation peak currents of catechol at Y2O3 (500 °C)/GCE and Y2O3 (800 °C)/GCE showed two stages linear function of concentration (2.0~20.0 × 10−6 mol/L, 20.0~60.0 × 10−6 mol/L). The results indicated that the detection limits were equal to 2.4 × 10−7 mol/L (Y2O3 (500 °C)/GCE) and 7.8 × 10−7 mol/L (Y2O3 (800 °C)/GCE). The study not only provided a method to synthesize metal oxide, but also proposed a promising on-line pH model to study cation exchange kinetics.

Measurement(s) Metabolite abundances in serum • Metabolite abundances in rumen fluid • taxonomic diversity assessment by targeted gene surveyin rumen fluid • metagenome in rumen fluid Technology Type(s) Liquid Chromatography Mass Spectrometry • next generation DNA sequencing • Next Generation Sequencing Factor Type(s) heat stress and growth stages Sample Characteristic - Organism Bos taurus Sample Characteristic - Environment growing heifers, heifers, and lactating cows were selected in April and August of Shandong Province

The synthesis of multielement composite oxide nanomaterials containing Ce, Zr, Y, and Yb was investigated using a micro confined jet mixer reactor operated in continuous mode under supercritical water conditions. The obtained nanoparticles were characterized using ICP-AES, SEM-EDS, FTIR, Raman spectroscopy, XRD, and TEM. All samples exhibited a uniform particle shape and a narrow particle size distribution. An analysis of the d-spacing results using selected electron area diffraction (SAED) patterns confirmed the production of cubic-phase crystals. A BET test was employed to determine the specific surface area of the prepared nanoparticles. OSC and TPR techniques were utilized to characterize the oxygen storage capacity and reduction performance of the obtained samples, with an analysis conducted to determine how the different proportions of elements affected the performance of multielement mixed oxides. The ionic conductivity of multielement composite oxide was measured using alternating current impedance spectroscopy (EIS), and the impact of Y, Ce, and Yb on the electrolyte material’s ionic conductivity was analyzed.

Abstract Objective To compare a new classification with the Garden classification by exploring their relationships with vascular injury. Methods This retrospective study enrolled 73 patients with subcapital femoral neck fracture from July 2015 to November 2018, including 32 males and 41 females with an average age of 47.2 years. All patients were classified by the Garden classification using anteroposterior X‐ray imaging and by a new classification system based on three‐dimensional CT imaging. The blood supply of the affected femoral head in these patients was evaluated based on DSA images. Correlations between the two classifications and the degree of vascular injury were assessed. Results The results of the DSA examination indicated that eight patients had no retinacular vessel injury, 20 patients had one retinacular vessel injury, 35 patients had two retinacular vessel injuries, and 10 patients had three retinacular vessel injuries. The degree of vascular injury was used to match the two fracture classifications. Forty‐nine Garden classifications (Type I‐IV: 8, 12, 23, 6, respectively, 67.12%) and 66 new classifications (Type I‐IV: 8, 20, 32, 6, respectively, 90.41%) corresponded to the degree of vascular injury (p

Antibody therapies have huge potential in treating cancer but currently suffer from problems with delivery. Here the authors report on a study into then conjugation of Ramucirumab to gold nanoparticles to improve delivery and add theranostic capabilities, demonstrating preferential toxicity in gastric cancer cells.

Yoon, Munoz, et al. investigate the rate of de novo coding and non-coding variants in families with high- and low-risk for autism using whole-genome sequence data from collections of families with autism. They demonstrate that de novo intronic variants increase the risk of autism, that the contribution of de novo variants is significantly larger in low-risk families, and that de novo variants contribute to 30-39% of cases of all autism.

Computational fluid dynamics (CFD) and population balance models (PBM) were coupled together for the first time to simulate the synthesis of zirconia nanoparticles in a continuous hydrothermal flow synthesis (CHFS) system with a self-designed confined impinging jet mixing (CJM) reactor. The hydrodynamic and thermodynamic behaviors within the CJM reactor strongly influenced the formation of the ZrO2 nanoparticles. Crucial parameters, such as velocities, temperatures, mixing conditions, and reaction rates, were analyzed under various supercritical conditions. Temperature and velocity measurements as functions of distance were also investigated. Normal particle size distribution (PSD) patterns were observed in all cases. The mean particle sizes in this study were calculated and compared using PBM aggregation analysis.

PurposeThe detection and characterization of focal liver lesions (FLLs) in patients with cirrhosis is challenging. Accurate information about FLLs is key to their management, which can range from conservative methods to surgical excision. We sought to develop a nomogram that incorporates clinical risk factors, blood indicators, and enhanced computed tomography (CT) imaging findings to predict the nature of FLLs in cirrhotic livers.MethodA total of 348 surgically confirmed FLLs were included. CT findings and clinical data were assessed. All factors with P < 0.05 in univariate analysis were included in multivariate analysis. ROC analysis was performed, and a nomogram was constructed based on the multivariate logistic regression analysis results.ResultsThe FLLs were either benign (n = 79) or malignant (n = 269). Logistic regression evaluated independent factors that positively affected malignancy. AFP (OR = 10.547), arterial phase hyperenhancement (APHE) (OR = 740.876), washout (OR = 0.028), satellite lesions (OR = 15.164), ascites (OR = 156.241), and nodule-in-nodule architecture (OR =27.401) were independent predictors of malignancy. The combined predictors had excellent performance in differentiating benign and malignant lesions, with an AUC of 0.959, a sensitivity of 95.24%, and a specificity of 87.5% in the training cohort and AUC of 0.981, sensitivity of 94.74%, and specificity of 93.33% in the test cohort. The C-index was 96.80%, and calibration curves showed good agreement between the nomogram predictions and the actual data.ConclusionsThe nomogram showed excellent discrimination and calibration for malignancy risk prediction, and it may aid in making FLLs treatment decisions.

In this paper, in order to improve the calculation accuracy and efficiency of α-order Caputo fractional derivative (0 < α ≤ 1), we developed a compact scheme combining the fast time stepping method for solving 2D fractional nonlinear subdiffusion equations. In the temporal direction, a time stepping method was applied. It can reach second-order accuracy. In the spatial direction, we utilized the compact difference scheme, which can reach fourth-order accuracy. Some properties of coefficients are given, which are essential for the theoretical analysis. Meanwhile, we rigorously proved the unconditional stability of the proposed scheme and gave the sharp error estimate. To overcome the intensive computation caused by the fractional operators, we combined a fast algorithm, which can reduce the computational complexity from O(N2) to O(Nlog(N)), where N represents the number of time steps. Considering that the solution of the subdiffusion equation is weakly regular in most cases, we added correction terms to ensure that the solution can achieve the optimal convergence accuracy.

Alzheimer’s disease (AD) is one of the most common neurodegenerative diseases with irreversible damage of the brain and a continuous pathophysiological process. Early detection and accurate diagnosis are essential for the early intervention of AD. Precise detection of blood biomarkers related to AD could provide a shortcut to identifying early-stage patients before symptoms. In recent years, targeting peptides or peptoids have been chosen as recognition elements in nano-sensors or fluorescence detection to increase the targeting specificity, while peptide-based probes were also developed considering their specific advantages. Peptide-based sensors and probes have been developed according to different strategies, such as natural receptors, high-throughput screening, or artificial design for AD detection. This review will briefly summarize the recent developments and trends of AD diagnosis platforms based on peptide and peptoid as recognition elements and provide insights into the application of peptide and peptoid with different sources and characteristics in the diagnosis of AD biomarkers.

Abstract Background Zinc finger proteins (ZFPs) containing only a single zinc finger domain play important roles in the regulation of plant growth and development, as well as in biotic and abiotic stress responses. To date, the evolutionary history and functions of the ZFP gene family have not been identified in cotton. Results In this paper, we identified 29 ZFP genes in Gossypium hirsutum. This gene family was divided into seven subfamilies, 22 of which were distributed over 17 chromosomes. Bioinformatic analysis revealed that 20 GhZFP genes originated from whole genome duplications and two originated from dispersed duplication events, indicating that whole genome duplication is the main force in the expansion of the GhZFP gene family. Most GhZFP8 subfamily genes, except for GhZFP8–3, were highly expressed during fiber cell growth, and were induced by brassinosteroids in vitro. Furthermore, we found that a large number of GhZFP genes contained gibberellic acid responsive elements, auxin responsive elements, and E-box elements in their promoter regions. Exogenous application of these hormones significantly stimulated the expression of these genes. Conclusions Our findings reveal that GhZFP8 genes are involved in cotton fiber development and widely induced by auxin, gibberellin and BR, which provides a foundation for the identification of more downstream genes with potential roles in phytohormone stimuli, and a basis for breeding better cotton varieties in the future.

Abstract Background The body is unable to repair and regenerate large area bone defects. Moreover, the repair capacity of articular cartilage is very limited. There has long been a lack of effective treatments for osteochondral lesions. The engineered tissue with biphase synthetic for osteochondral repair has become one of the hot research fields over the past few years. In this study, an integrated biomanufacturing platform was constructed with bone marrow mesenchymal stem cells (BMSCs)/porous tantalum (pTa) associated with chondrocytes/collagen membranes (CM) to repair large osteochondral defects in load-bearing areas of goats. Methods Twenty-four goats with a large osteochondral defect in the femoral heads of the left hind legs were randomly divided into three groups: eight were treated with chondrocytes/CM-BMSCs/pTa, eight were treated with pure CM-pTa composite, and the other eight goats were untreated. The repair effect was assessed by X-ray, gross observation, and histomorphology for 16 weeks after the operation. In addition, the biocompatibility of chondrocytes/CM-BMSCs/pTa was observed by flow cytometry, CCK8, immunocytochemistry, and Q-PCR. The characteristics of the chondrocytes/CM-BMSCs/pTa were evaluated using both scanning electron microscopy and mechanical testing machine. Results The integrated repair material consists of pTa, injectable fibrin sealant, and CM promoted adhesion and growth of BMSCs and chondrocytes. pTa played an important role in promoting the differentiation of BMSCs into osteoblasts. Three-dimensional CM maintained the phenotype of chondrocytes successfully and expressed chondrogenic genes highly. The in vivo study showed that after 16 weeks from implantation, osteochondral defects in almost half of the femoral heads had been successfully repaired by BMSC-loaded pTa associated with biomimetic 3D collagen-based scaffold. Conclusions The chondrocytes/CM-BMSCs/pTa demonstrated significant therapeutic efficacy in goat models of large osteochondral defect. This provides a novel therapeutic strategy for large osteochondral lesions in load-bearing areas caused by severe injury, necrosis, infection, degeneration, and tumor resection with a high profile of safety, effectiveness, and simplicity.

Protein–polysaccharide complexes, which involve Maillard-type protein–polysaccharide conjugates and electrostatic protein–polysaccharide complexes, have the potential to stabilize oleogel-based nanoemulsions for nutraceutical delivery. Here, ovalbumin (OVA) and gum arabic (GA) were used to prepare OVA–GA conjugate (OGC) and OVA–GA mixture (OGM), followed by the fabrication of astaxanthin-loaded oleogel-based nanoemulsions. Carnauba wax (5% w/w) and rice bran oil were mixed to prepare food-grade oleogel. The successful preparation of OGC was verified by means of SDS-PAGE analysis and free amino groups determination. OGC endowed oleogel-based nanoemulsions with smaller emulsion droplets and higher stability during 30-day storage, implying more outstanding emulsifying capability than OGM. Both OGC-stabilized nanoemulsions and OGM-stabilized nanoemulsions could enhance the extent of lipolysis and the bioaccessibility of astaxanthin compared with oleogel. Meanwhile, OGC exhibited significantly better than OGM, which indicated that OGC-stabilized oleogel-based nanoemulsions possessed more desirable nutraceutical delivery performance than OGM-stabilized oleogel-based nanoemulsions. This study may fill a gap in the influence of different protein–polysaccharide complexes on oleogel-based nanoemulsions and contribute to deeper insights about novel oleogel-based nanoemulsions for their applications in the food industry.

Exosomes are naturally occurring nanoscale vesicles that are released and received by almost all cells in the body. Exosomes can be transferred between cells and contain various molecular constitutes closely related to their origin and function, including proteins, lipids, and RNAs. The importance of exosomes in cellular communication makes them important vectors for delivering a variety of drugs throughout the body. Exosomes are ubiquitous in the circulatory system and can reach the site of injury or disease through a variety of biological barriers. Due to its unique structure and rich inclusions, it can be used for the diagnosis and treatment of diseases. Mesenchymal stem-cell-derived exosomes (MSCs-Exo) inherit the physiological functions of MSCs, including repairing and regenerating tissues, suppressing inflammatory responses, and regulating the body’s immunity; therefore, MSCs-Exo can be used as a natural drug delivery carrier with therapeutic effects, and has been increasingly used in the treatment of cardiovascular diseases and neurodegenerative diseases. Here, we summarize the research progress of MSCs-Exo as drug delivery vectors and their application for various drug deliveries, providing ideas and references for the study of MSCs-Exo in recent years.

Cancer stem cells (CSCs) are a small subset of cells that sit atop the hierarchical ladder in many cancer types. Liver CSCs have been associated with high chemoresistance and recurrence rates in hepatocellular carcinoma (HCC). However, as of yet, no satisfactorily effective liver CSC-targeted treatment is available, which drove us to design and investigate the efficacy of a liposome-based delivery system. Here, we introduce a redox-triggered dual-targeted liposome, CEP-LP@S/D, capable of co-delivering doxorubicin (Dox) and salinomycin (Sal) for the synergistic treatment of liver cancer. This system is based on the association of CD133- and EpCAM-targeted peptides to form Y-shaped CEP ligands that were anchored to the surface of the liposome and allowed the selective targeting of CD133+ EpCAM+ liver CSCs. After arriving to the CSCs, the CEP-LP@S/D liposome undergoes endocytosis to the cytoplasm, where a high concentration of glutathione (GSH) breaks its disulfide bonds, thereby degrading the liposome. This then induces a rapid release of Dox and Sal to synergistically inhibit tumor growth. Notably, this effect occurs through Dox-induced apoptosis and concurrent lysosomal iron sequestration by Sal. Interestingly, both in vitro and in vivo studies indicated that our GSH-responsive co-delivery system not only effectively enhanced CSC targeting but also eliminated the non-CSC faction, thereby exhibiting high antitumor efficacy. We believe that the smart liposome nanocarrier-based co-delivery system is a promising strategy to combat liver cancer, which may also lay the groundwork for more enhanced approaches to target other cancer types as well.

A confined jet mixing reactor operated in continuous hydrothermal flow synthesis was investigated for the synthesis of CeO2-ZrO2 (CZ) nanoparticles. The obtained ultrafine powders were characterized using scanning electron microscopy–energy dispersive spectrometry (SEM-EDS), inductively coupled plasma–atomic emission spectroscopy (ICP-AES), Fourier transform infrared spectroscopy (FTIR), Raman spectroscopy, X-ray diffraction analysis (XRD), transmission electron microscopy (TEM) coupled with selected area electron diffraction (SAED), a BET (Brunauer-Emmett-Teller)-specific surface area test and pore analysis, oxygen storage capacity (OSC) test, and a H2 temperature programmed reduction (H2-TPR) test. The XRD results show that all samples were composed of high-purity cubic CZ nanoparticles. High resolution transmission electron microscope (HR-TEM) analysis showed that CZ nanoparticles with uniform size and shape distributions were obtained in this investigation. The d-spacing values, determined based on the TEM-selected area electron diffraction (SAED) patterns, were in good agreements with the reference data. BET results showed that the prepared CZ samples had large specific surface areas. Pore volume and size distribution were obtained by pore analysis. Oxygen pulse adsorption technology was used to test the oxygen storage capacity of the sample. The redox capacity of the CZ material was determined by a H2 temperature-programmed reduction test.

Abstract EGFR mutations companion diagnostics have been proved to be crucial for the efficacy of tyrosine kinase inhibitor targeted cancer therapies. To uncover multiple mutations occurred in minority of EGFR-mutated cells, which may be covered by the noises from majority of un-mutated cells, is currently becoming an urgent clinical requirement. Here we present the validation of a microfluidic-chip-based method for detecting EGFR multi-mutations at single-cell level. By trapping and immunofluorescently imaging single cells in specifically designed silicon microwells, the EGFR-expressed cells were easily identified. By in situ lysing single cells, the cell lysates of EGFR-expressed cells were retrieved without cross-contamination. Benefited from excluding the noise from cells without EGFR expression, the simple and cost-effective Sanger’s sequencing, but not the expensive deep sequencing of the whole cell population, was used to discover multi-mutations. We verified the new method with precisely discovering three most important EGFR drug-related mutations from a sample in which EGFR-mutated cells only account for a small percentage of whole cell population. The microfluidic chip is capable of discovering not only the existence of specific EGFR multi-mutations, but also other valuable single-cell-level information: on which specific cells the mutations occurred, or whether different mutations coexist on the same cells. This microfluidic chip constitutes a promising method to promote simple and cost-effective Sanger’s sequencing to be a routine test before performing targeted cancer therapy.

Vesicoureteral reflux (VUR) is a complex, heritable disorder. Genome-wide linkage analyses of families affected by VUR have revealed multiple genomic loci linked to VUR. These loci normally harbor a number of genes whose biologically functional variant is yet to be identified. DNA copy number variations (CNVs) have not been extensively studied at high resolution in VUR patients. In this study, we performed array comparative genomic hybridization (aCGH) on a cohort of patients with a history of both VUR and urinary tract infection (UTI) with the objective of identifying genetic variations responsible for VUR and/or UTI susceptibility. UTI/VUR-associated CNVs were identified by aCGH results from the 192 Randomized Intervention for Children With Vesicoureteral Reflux (RIVUR) patients compared to 683 controls. Rare, large CNVs that are likely pathogenic and lead to VUR development were identified using stringent analysis criteria. Because UTI is a common affliction with multiple risk factors, we utilized standard analysis to identify potential disease-modifying CNVs that can contribute to UTI risk. Gene ontology analysis identified that CNVs in innate immunity and development genes were enriched in RIVUR patients. CNVs affecting innate immune genes may contribute to UTI susceptibility in VUR patients and may provide the first step in assisting clinical medicine in determining adverse outcome risk in children with VUR.

In this paper, a novel combined heat and power (CHP) system is proposed in which the waste heat from a supercritical CO2 recompression Brayton cycle (sCO2) is recovered by a LiBr-H2O absorption heat pump (AHP). Thermodynamic and exergoeconomic models are established on the basis of the mass, energy, and cost balance equations. The proposed sCO2/LiBr-H2O AHP system is examined and compared with a stand-alone sCO2 system, a sCO2/DH system (sCO2/direct heating system), and a sCO2/ammonia-water AHP system from the viewpoints of energy, exergy, and exergoeconomics. Parametric studies are performed to reveal the influences of decision variables on the performances of these systems, and the particle swarm optimization (PSO) algorithm is utilized to optimize the system performances. Results show that the sCO2/LiBr-H2O AHP system can obtain an improvement of 13.39% in exergy efficiency and a reduction of 8.66% in total product unit cost compared with the stand-alone sCO2 system. In addition, the sCO2/LiBr-H2O AHP system performs better than sCO2/DH system and sCO2/ammonia-water AHP system do, indicating that the LiBr-H2O AHP is a preferable bottoming cycle for heat production. The detailed parametric analysis, optimization, and comparison results may provide some references in the design and operation of sCO2/AHP system to save energy consumption and provide considerable economic benefits.

Here, we provide direct evidence that using recombinant proteins expressed in eukaryotic cells as antigen is a practical way to generate monoclonal antibodies (mAbs) against heavily glycosylated proteins. Heavily glycosylated proteins are typically difficult targets for mAb generation, being limited by unsatisfactory affinity and low specificity. Using the heavily glycosylated CD45 protein as an example, we demonstrate the entire process of expressing the protein in eukaryotic cells and using it as an antigen to generate CD45-targeting mAbs in mice. The mAbs generated showed robust affinity and specificity, which are crucial factors for differentiate circulating tumor cells from white blood cells in human breast cancer patient samples. Only 1 cell fusion and 2 cyclic sub-cloning steps were necessary before mAbs with satisfactory performance were obtained.

A high level of HER2 expression in breast cancer correlates with a higher tumor growth rate, high metastatic potential, and a poor long-term patient survival rate. Pertuzumab, a human monoclonal antibody, can reduce the effect of HER2 overexpression by preventing HER2 dimerization. In this study, a combination protocol of molecular dynamics modeling and MM/GBSA binding free energy calculations was applied to design peptides that interact with HER2 based on the HER2/pertuzumab crystal structure. Based on a β hairpin in pertuzumab from Glu46 to Lys65-which plays a key role in interacting with HER2-mutations were carried out in silico to improve the binding free energy of the hairpin that interacts with the Phe256-Lys314 of the HER2 protein. Combined the use of one-bead-one-compound library screening, among all the mutations, a peptide (58F63Y) with the lowest binding free energy was confirmed experimentally to have the highest affinity, and it may be used as a new probe in diagnosing and treating HER2-positive breast cancer.

Determining therapeutic efficacy is critical for tumor precision theranostics. In order to monitor the efficacy of anti-cancer drugs (e.g., Paclitaxel), a pH-sensitive ratiometric fluorescent imaging probe was constructed. The pH-sensitive ratiometric fluorescent dye ANNA was covalently coupled to the N-terminal of the cell-penetrating TAT peptide through an amidation reaction (TAT-ANNA). The in vitro cellular experiments determined that the TAT-ANNA probe could penetrate the cell membrane and image the intracellular pH in real time. The in vivo experiments were then carried out, and the ratiometric pH response to the state of the tumor was recorded immediately after medication. The TAT-ANNA probe was successfully used to monitor the pharmacodynamics of anti-cancer drugs in vivo.

The use of graphene has leapt forward the materials field and the functional modification of graphene has not stopped. In this work, β-lactoglobulin (BLG) was used to functionalize reduced graphene oxide (RGO) based on its amphiphilic properties. Also, trimetallic PtAuPd nanoparticles were reduced to the surface of BLG-functionalized RGO and formed BLG-PtAuPd-RGO nanocomposite using facile synthesis. Transmission electron microscopy, energy-dispersive X-ray spectroscopy and Fourier transform infrared spectra were used to characterize the nanocomposite. Electrocatalytic analysis was evaluated through cyclic voltammetry and chronoamperometry methods. We developed a glucose sensor by fabricating GOD-BLG-PtAuPd-RGO/glassy carbon (GC) electrode. It presented a remarkable sensitivity of 63.29 μA mM−1 cm−2 (4.43 μA mM−1), a wider linear range from 0.005 to 9 mM and a lower detection limit of 0.13 μM (S/N = 3). Additionally, the glucose sensor exhibited excellent testing capability in human serum samples.

Liver cirrhosis is a potentially life-threatening disease caused by progressive displacement of functional hepatocytes by fibrous tissue. The underlying fibrosis is often driven by chronic infection with hepatitis B virus (HBV). Matrix metalloproteinases including MMP-8 are crucial for excess collagen degradation. In a rat model of liver cirrhosis, MMP-8 delivery by an adenovirus (Ad) vector achieved significant amelioration of fibrosis but application of Ad vectors in humans is subject to various issues, including a lack of intrinsic liver specificity.HBV is highly liver-specific and its principal suitability as liver-specific gene transfer vector is established. HBV vectors have a limited insertion capacity and are replication-defective. Conversely, in an HBV infected cell vector replication may be rescued in trans by the resident virus, allowing conditional vector amplification and spreading. Capitalizing on a resident pathogen to help in its elimination and/or in treating its pathogenic consequences would provide a novel strategy. However, resident HBV may also reduce susceptibility to HBV vector superinfection. Thus a size-compatible truncated MMP-8 (tMMP8) gene was cloned into an HBV vector which was then used to generate a chimeric Ad-HBV shuttle vector that is not subject to superinfection exclusion. Rats with thioacetamide-induced liver cirrhosis were injected with the chimera to evaluate therapeutic efficacy.Our data demonstrate that infectious HBV vector particles can be obtained via trans-complementation by wild-type virus, and that the tMMP8 HBV vector can efficiently be shuttled by an Ad vector into cirrhotic rat livers. There it exerted a comparable beneficial effect on fibrosis and hepatocyte proliferation markers as a conventional full-length MMP-8Ad vector.Though the rat cirrhosis model does not allow assessing in vivo HBV vector amplification these results advocate the further development of Ad-HBV vectors for liver-specific gene therapy, including and perhaps particularly for HBV-related disease.

Viral vectors are engineered virus variants able to deliver nonviral genetic information into cells, usually by the same routes as the parental viruses. For several virus families, replication-competent vectors carrying reporter genes have become invaluable tools for easy and quantitative monitoring of replication and infection, and thus also for identifying antivirals and virus susceptible cells. For hepatitis B virus (HBV), a small enveloped DNA virus causing B-type hepatitis, such vectors are not available because insertions into its tiny 3.2 kb genome almost inevitably affect essential replication elements. HBV replicates by reverse transcription of the pregenomic (pg) RNA which is also required as bicistronic mRNA for the capsid (core) protein and the reverse transcriptase (Pol); their open reading frames (ORFs) overlap by some 150 basepairs. Translation of the downstream Pol ORF does not involve a conventional internal ribosome entry site (IRES). We reasoned that duplicating the overlap region and providing artificial IRES control for translation of both Pol and an in-between inserted transgene might yield a functional tricistronic pgRNA, without interfering with envelope protein expression. As IRESs we used a 22 nucleotide element termed Rbm3 IRES to minimize genome size increase. Model plasmids confirmed its activity even in tricistronic arrangements. Analogous plasmids for complete HBV genomes carrying 399 bp and 720 bp transgenes for blasticidin resistance (BsdR) and humanized Renilla green fluorescent protein (hrGFP) produced core and envelope proteins like wild-type HBV; while the hrGFP vector replicated poorly, the BsdR vector generated around 40% as much replicative DNA as wild-type HBV. Both vectors, however, formed enveloped virions which were infectious for HBV-susceptible HepaRG cells. Because numerous reporter and effector genes with sizes of around 500 bp or less are available, the new HBV vectors should become highly useful tools to better understand, and combat, this important pathogen.

In the title molecule, C24H28N6O2S2, the dihedral angle between the aromatic ring planes is 42.2 (1)°. In the crystal structure, the hydroxy groups are involved in O—H...S hydrogen bonding, which links the molecules into corrugated layers propagating parallel to the bc plane.