Proteomic, single-cell and bulk transcriptomic analysis of plasma and tumor tissues unveil core proteins in response to anti-PD-L1 immunotherapy in triple negative breast cancer.

BACKGROUND: Anti-PD-1/PD-L1 treatment has achieved durable responses in TNBC patients, whereas a fraction of them showed non-sensitivity to the treatment and the mechanism is still unclear. METHODS: Pre- and post-treatment plasma samples from triple negative breast cancer (TNBC) patients treated with immunotherapy were measured by tandem mass tag (TMT) mass spectrometry. Public proteome data of lung cancer and melanoma treated with immunotherapy were employed to validate the findings. Blood and tissue single-cell RNA sequencing (scRNA-seq) data of TNBC patients treated with or without immunotherapy were analyzed to identify the derivations of plasma proteins. RNA-seq data from IMvigor210 and other cancer types were used to validate plasma proteins in predicting response to immunotherapy. RESULTS: A random forest model constructed by FAP, LRG1, LBP and COMP could well predict the response to immunotherapy. The activation of complement cascade was observed in responders, whereas FAP and COMP showed a higher abundance in non-responders and negative correlated with the activation of complements. scRNA-seq and bulk RNA-seq analysis suggested that FAP, COMP and complements were derived from fibroblasts of tumor tissues. CONCLUSIONS: We constructe an effective plasma proteomic model in predicting response to immunotherapy, and find that FAP+ and COMP+ fibroblasts are potential targets for reversing immunotherapy resistance.

Adoptive cell immunotherapy for breast cancer: harnessing the power of immune cells.

Breast cancer is the most prevalent malignant neoplasm worldwide, necessitating the development of novel therapeutic strategies owing to the limitations posed by conventional treatment modalities. Immunotherapy is an innovative approach that has demonstrated significant efficacy in modulating a patient's innate immune system to combat tumor cells. In the era of precision medicine, adoptive immunotherapy for breast cancer has garnered widespread attention as an emerging treatment strategy, primarily encompassing cellular therapies such as tumor-infiltrating lymphocyte therapy, chimeric antigen receptor T/NK/M cell therapy, T-cell receptor gene-engineered T-cell therapy, lymphokine-activated killer cell therapy, cytokine-induced killer cell therapy, natural killer cell therapy, and γδ T cell therapy, among others. This treatment paradigm is based on the principles of immune memory and antigen specificity, involving the collection, processing, and expansion of the patient's immune cells, followed by their reintroduction into the patient's body to activate the immune system and prevent tumor recurrence and metastasis. Currently, multiple clinical trials are assessing the feasibility, effectiveness, and safety of adoptive immunotherapy in breast cancer. However, this therapeutic approach faces challenges associated with tumor heterogeneity, immune evasion, and treatment safety. This review comprehensively summarizes the latest advancements in adoptive immunotherapy for breast cancer and discusses future research directions and prospects, offering valuable guidance and insights into breast cancer immunotherapy.

Quercetin protects against Aspergillus fumigatus keratitis by reducing fungal load and inhibiting TLR-4 induced inflammatory response.

PURPOSE: To investigate the antifungal and anti-inflammatory effects of quercetin in Aspergillus fumigatus (A. fumigatus) keratitis. METHODS: Draize eye test was performed in mice to evaluate the toxicity of quercetin, and the antifungal effects on A. fumigatus were assessed via scanning electron microscopy (SEM), propidium iodide uptake, and adherence assay. In fungal keratitis (FK) mouse models, immunostaining was performed for investigating toll-like receptor 4 (TLR-4) expression and macrophage infiltration. Real-time PCR, ELISA, and Western blot were used to evaluate the expression of pro-inflammatory factors IL-1ß, TNF-α, and IL-6 in infected RAW264.7 cells. Cells were also treated with TLR-4 siRNA or agonist CRX-527 to investigate mechanisms underlying the anti-inflammatory activity of quercetin. RESULTS: Quercetin at 32 µM was non-toxic to corneal epithelial and significantly inhibited A. fumigatus growth and adhesion, and also altered the structure and reduced the number of mycelia. Quercetin significantly reduced macrophage infiltration in the mouse cornea, and attenuated the expression of TLR-4 in the corneal epithelium and stroma of mice with keratitis caused by A. fumigatus. In RAW264.7 cells infected by A. fumigatus, quercetin downregulated TLR-4 along with pro-inflammatory factors IL-1ß, TNF-α, and IL-6. RAW cells with TLR-4 knockdown had reduced expression of factors after A. fumigatus infection, which was decreased even further with quercetin treatment. In contrast, cells with CRX-527 had elevated inflammatory factors compared to control, which was significantly attenuated in the presence of quercetin. CONCLUSION: Quercetin plays a protective role in mouse A. fumigatus keratitis by inhibiting fungal load, disrupting hyphae structure, macrophage infiltration, and suppressing inflammation response in macrophages via TLR-4 mediated signaling pathway.

AMPK-FOXO-IP3R signaling pathway mediates neurological and developmental defects caused by mitochondrial DNA mutations.

Pathological mutations in human mitochondrial genomes (mtDNA) can cause a series of neurological, behavioral, and developmental defects, but the underlying molecular mechanisms are poorly understood. We show here that the energy-sensing adenosine monophosphate (AMP)-activated protein kinase (AMPK) signaling pathway plays a key role in mediating similar defects caused by different mtDNA mutations in Caenorhabditis elegans, including loss or reduction of osmotic, chemical and olfactory sensing, locomotion, and associative learning and memory, as well as increased embryonic lethality. mtDNA mutations cause reduced ATP (adenosine triphosphate) levels, activation of C. elegans AMPK AAK-2, and nuclear translocation of the FOXO transcription factor DAF-16. Activated DAF-16 up-regulates the expression of inositol triphosphate receptor ITR-1, an endoplasmic reticulum calcium channel, leading to increased basal cytosolic Ca2+ levels, decreased neuronal responsiveness, compromised synapses, and increased embryonic death. Treatment of mtDNA mutants with vitamin MK-4 restores cellular ATP and cytosolic Ca2+ levels, improves synaptic development, and suppresses sensory and behavioral defects and embryonic death. Our study provides crucial mechanistic insights into neuronal and developmental defects caused by mtDNA mutations and will improve understanding and treatment of related mitochondrial diseases.

Hamming Distance Optimized Underwater Acoustic OTFS-IM Systems.

The orthogonal time frequency space (OTFS) modulation technique can provide reliable communication in time-varying channels. Due to the dispersive characteristics of underwater acoustic channels, this paper proposes an OTFS-IM underwater acoustic communication system based on Hamming distance optimization to reduce the impact of dispersion in underwater acoustic communication. Firstly, the OTFS-IM underwater acoustic communication system is introduced, which introduces index modulation into the Delay-Doppler (DD) domain to make the OTFS system have stronger anti-Delay-Doppler capability. In contrast, since there is index sequence redundancy in a specific index combination, a Hamming distance optimization model is used to eliminate the redundant combination in the specific index combination sequence and further improve the bit error rate performance of the system. In addition, the Hamming distance optimized OTFS-IM underwater acoustic communication system is verified by simulation analysis. The results show that the proposed Hamming distance optimized OTFS-IM can achieve more reliable bit error rate performance.

Construction and hydrophilic modification of dual-network structured nonwoven/UHMWPE composite membranes for water processing.

Water pollution caused by the continuous development of industrialization has always been a common concern of mankind. Herein, a novel strategy to fabricate a high-performance composite membrane based on dual-network structured nonwoven net/UHMWPE nanopores via a thermal phase separation and composite technique is reported. By thermal phase separation of ultra-high-molecular weight polyethylene (UHMWPE)/liquid paraffin (LP), this approach enables 3D nanopores to tightly bond with a nonwoven net to form a dual-network structure. The dual-network composite membrane possesses the integrated features of pore structure and high porosity (89.9%). After modification with hyperbranched polymers (HBPs), the composite membrane with the desirable surface chemistry achieves high-efficiency filtration (water flux = 1054 L m-2 h-1, rejection rate = 50 nm PS nanospheres almost close to 100%, and antibacterial properties). The fabrication of such composites may provide new insights into the design and development of high-performance filtration and separation materials for various applications.

Low-carbon and high-ammonia nitrogen dispersed wastewater treatment: From "normal-sludge" to "low-sludge" to "no-sludge" modes.

Several biological enhancements were implemented in the aerobic tank to address the challenges of treating expressway service sewage (ESS) with low-carbon and high-ammonia nitrogen using A/O-MBR technology, aiming to improve TN removal efficiency and reduce excessive sludge production. A novel moving bed biofilm reactor (MBBR) inoculated with heterotrophic nitrification-aerobic denitrification (HN-AD) bacteria was developed for ESS, and the results showed that HN-AD bacteria significantly improved TN removal efficiency, with an increase of 65% compared to the traditional activated sludge system. High-throughput sequencing revealed that Bacteroidotas contributed significantly to MBBR denitrification, and the genes nirK and nosZ played a significant role in denitrification. The HN-AD biofilm-forming MBBR achieved the transition of ESS treatment from "normal-sludge" mode to the more environmentally-friendly "low-sludge" and "no-sludge" modes by reducing the sludge concentration.

Neratinib inhibits proliferation and promotes apoptosis of acute myeloid leukemia cells by activating autophagy-dependent ferroptosis.

Acute myeloid leukemia (AML) is a hematologic malignancy with increased lethality. We focused on elucidating the role of Neratinib, a tyrosine kinase inhibitor, in the progression of AML and identify the potential mechanisms. Upon the treatment of Neratinib, autophagy suppressor 3-methyladenine (3-MA) and ferroptosis stimulator Erastin, the viability and proliferation of HL-60 cells were evaluated by cell counting kit-8 and 5-Ethynyl-20-Deoxyuridine staining assays. A flow cytometer was to observe cell cycle and apoptosis. Production of reactive oxygen species (ROS) was tested via 2,7-dichlorodihydrofluorescein diacetate  assay. Additionally, malondialdehyde (MDA) content and Fe2+ activity were examined with commercial kits. LC3-II expression was examined by using immunofluoresence staining. Western blot analysis ascertained the expression of proliferation, apoptosis, ferroptosis and autophagy-associated proteins. It was noted that Neratinib notably mitigated cell viability and proliferation, cut down Ki67 and proliferating cell nuclear antigen expression. Moreover, Neratinib hindered cell cycle at G0/G1 phase whereas exacerbated apoptosis. ROS, MDA and Fe2+ activities were elevated by Neratinib, coupled with the reduced glutathione peroxidase 4, ferritin heavy chain 1 expression and enhanced acyl-CoA synthetase long-chain family member 4 expression. Furthermore, Neratinib promoted autophagy of HL-60 cells, evidenced by raised LC3-II, ATG5, Beclin1 expression and lessened p62 expression. Importantly, 3-MA eased the impacts of Neratinib on cell ferroptosis, proliferation and apoptosis, which were offset by further administration of Erastin. To conclude, Neratinib could suppress proliferation and promote apoptosis of HL-60 cells through autophagy-dependent ferroptosis.

Nitrogen removal and mechanism of an extremely high-ammonia tolerant heterotrophic nitrification-aerobic denitrification bacterium Alcaligenes faecalis TF-1.

A novel heterotrophic nitrifying bacterium with high salt and high ammonia nitrogen tolerance, Alcaligenes faecalis TF-1, was isolated from the leachate of a landfill. The verification of nitrogen removal efficiency of different nitrogen sources and PCR amplification electrophoresis results showed that the HN-AD pathway of the strain TF-1 was NH4+ â†’ NH2OH â†’ NO â†’ N2O â†’ N2. The results of parameter optimization showed that the optimal nitrogen removal conditions were as follows: sodium citrate as carbon source, C/N = 16, pH = 7, and NH4+-N loading of 808.21 mg/L. The strain TF-1 could remove about 94.60% of ammonia nitrogen (1963.94 mg/L). The salinity tolerance range of the strain TF-1 was 0-70 g/L, and the removal efficiency was 52.87% at salinity 70 g/L and NH4+-N concentration 919.20 mg/L and 55.67% at pH = 10 and NH4+-N concentration 994.82 mg/L. The extreme environmental adaptability and remarkable nitrogen removal performance make this strain a promising candidate in leachate treatment.

Bioaugmentation with Acinetobacter sp. TAC-1 to enhance nitrogen removal in swine wastewater by moving bed biofilm reactor inoculated with bacteria.

To enhance the performance of moving bed biofilm reactor (MBBR) inoculated with heterotrophic nitrification-aerobic denitrification (HN-AD) bacteria, bioaugmentation with Acinetobacter sp. TAC-1 was firstly employed and then the treatment performance for real swine wastewater was presented in this study. Results indicated that NH4+-N and TN removal rates of bioaugmented reactor were significantly improved from 16.53 mg/L/h and 16.15 mg/L/h to 24.58 mg/L/h and 24.45 mg/L/h, respectively. The efficient removal performance (NH4+-N 95.01%, TN 86.40%) for real swine wastewater was achieved within 24 h. Microbial analysis indicated that the composition of functional bacteria varied with the introduction of Acinetobacter sp. TAC-1, especially the abundance of Acinetobacter, Paracoccus and Rhodococcus related to the nitrogen removal. Furthermore, bioaugmentation with Acinetobacter sp. TAC-1 increased abundance of enzymes and functional genes (nirS, nirK and norZ) corresponding to denitrification that may be responsible for the enhanced nitrogen removal performance.

The influence mechanism of DO on the microbial community and carbon source metabolism in two solid carbon source systems.

The regulation mechanism of parameters on microorganisms and carbon source metabolism of solid carbon source simultaneous nitrification and denitrification (SND) process is not clear. In this paper, the effects of dissolved oxygen (DO) and biodegradable polymer (BDPs) types ((Polycaprolactone, PCL) and (Polybutylene succinate, PBS)) on treatment performance and microbial characteristics were investigated. The results show that the total nitrogen (TN) removal efficiency of SND process using PBS and PCL as fillers reached 93.02% and 97.28% under optimal parameter of DO 5 mg/L, respectively. The dominant genus with nitrogen removal performance in the PCL carbon source system are Hydrogenophaga and Acidovorax, and the main genus in the PBS system are Acidovorax and unclassified_Comamonadaceae. The co-metabolic network in PCL is more complex and easier to be regulated by DO. The BDPs types mainly affect the co-metabolic network with nodes of Thiothrix and Chryseomicrobium, ultimately leading to changes in the community structure. By comparison, BDPs types have a more significant impact on community structure than DO under low DO conditions (1 and 2 mg/L), but not under high DO condition(5 mg/L). Further, the distribution of functional enzymes may conflict between nitrification and carbon source degradation under high DO condition. Controlling the DO within the range of 2 mg-5 mg can further improve carbon source utilization efficiency.

Response of rotating biological contactor started up by heterotrophic nitrification-aerobic denitrification bacteria to various C/N ratios.

To improve the low pollutant removal efficiency of traditional biological methods for treating livestock and poultry breeding wastewater under a relatively low temperature, a rotating biological contactor (RBC) inoculated with heterotrophic nitrification-aerobic denitrification (HN-AD) bacteria was designed. A quick start-up process and a well removal performance had been achieved in the novel RBC. To elucidate the anti-load shock ability of the novel RBC, the effects of C/N ratio on nitrogen removal and microbial assemblage were focused under a low temperature (12 ± 2 °C). Results showed that the highest NH4+-N and TN removal efficiency were 99.57 ± 0.31% and 68.41 ± 0.52%. Microbial diversity analysis based on high throughput sequencing technique showed that Arcobacter and Flavobacterium with an increasing relative abundance were the key to ensure high nitrogen removal efficiently at a low C/N ratio and temperature. Moreover, nitrogen transferring pathways of the novel RBC was revealed and dissimilatory nitrate reduction and denitrification were the main pathways. The excellent pollutant removal performance demonstrates that the novel RBC is a promising process to effectively treat wastewater with low C/N ratio and low temperature.

Ship Radiated Noise Recognition Technology Based on ML-DS Decision Fusion.

Ship radiated noise is an important information source of underwater acoustic targets, and it is of great significance to the identification and classification of ship targets. However, there are a lot of interference noises in the water, which leads to the reduction of the model recognition rate. Therefore, the recognition results of radiated noise targets are severely affected. This paper proposes a machine learning Dempster-Shafer (ML-DS) decision fusion method. The algorithm combines the recognition results of machine learning and deep learning. It uses evidence-based decision-making theory to realize feature fusion under different neural network classifiers and improve the accuracy of judgment. First, deep learning algorithms are used to classify two-dimensional spectrogram features and one-dimensional amplitude features extracted from CNN and LSTM networks. The machine learning algorithm SVM is used to classify the chromaticity characteristics of radiated noise. Then, according to the classification results of different classifiers, a basic probability assignment model (BPA) was designed to fuse the recognition results of the classifiers. Finally, according to the classification characteristics of machine learning and deep learning, combined with the decision-making of D-S evidence theory of different times, the decision-making fusion of radiated noise is realized. The results of the experiment show that the two fusions of deep learning combined with one fusion of machine learning can significantly improve the recognition results of low signal-to-noise ratio (SNR) datasets. The lowest fusion recognition result can reach 76.01%, and the average fusion recognition rate can reach 94.92%. Compared with the traditional single feature recognition algorithm, the recognition accuracy is greatly improved. Compared with the traditional one-step fusion algorithm, it can effectively integrate the recognition results of heterogeneous data and heterogeneous networks. The identification method based on ML-DS proposed in this paper can be applied in the field of ship radiated noise identification.

Effect of bacteria-to-algae volume ratio on treatment performance and microbial community of a novel heterotrophic nitrification-aerobic denitrification bacteria-chlorella symbiotic system.

A novel symbiotic system combined by heterotrophic nitrification-aerobic denitrification (HN-AD) mixed bacteria and Chlorella pyrenoidosa was firstly proposed to resolve the poor tolerance and nitrogen removal performance of traditional symbiotic system for treating high ammonia biogas slurry. Results showed that the volume ratio of bacteria to algae had significant effects on nitrogen removal efficiency, microbial community structure, functional bacteria and genes. The optimal ratio was 1/3, and the average removal efficiency of TN and TP increased by 28.9% and 67.6% respectively, compared to those of HN-AD bacteria. High-throughput sequencing indicated nitrogen removal was jointly completed by HN-AD and heterotrophic denitrification. HN-AD bacteria Halomonas and Pseudomonas played a key role in nitrogen removal, and Rhodocyclaceae and Paracoccus took an important part in phosphorus removal. According to the functional gene prediction, the total relative abundance of nitrogen removal genes (0.0127%) and narG, narH and narL genes (0.0054%) were highest in 1/3 system.

Interaction of interregional O3 pollution using complex network analysis.

In order to improve the accuracy of air pollution management and promote the efficiency of coordinated inter-regional prevention and control, this study analyzes the interaction of O3 in Qilihe District, Lanzhou City, China. Data used for analysis was obtained from 63 air quality monitoring stations between November 2017 and October 2018. This paper uses complex network theory to describe the network structure characteristics of O3 pollution spatial correlation. On this basis, the node importance method is used to mine the sub-network with the highest spatial correlation in the O3 network, and use transfer entropy theory to analyse the interaction of pollutants between regions. The results show that the O3 area of Qilihe District, Lanzhou City can be divided into three parts: the urban street community type areas in urban areas, the township and village type areas in mountain areas and the scattered areas represented by isolated nodes. An analysis of the mutual influence of O3 between each area revealed that the impact of O3 on each monitoring station in adjacent areas will vary considerably. Therefore these areas cannot be governed as a whole, and the traditional extensive management measures based on administrative divisions cannot be used to replace all other regional governance measures. There is the need to develop a joint prevention and control mechanism tailored to local conditions in order to improve the accuracy and efficiency of O3 governance.

Efficiency and microbial diversity of aeration solid-phase denitrification process bioaugmented with HN-AD bacteria for the treatment of low C/N wastewater.

To evaluate the simultaneous nitrification and denitrification (SND) performance of the aeration solid-phase denitrification (SPD) process and improve the operating efficiency, aeration SPD process using polybutanediol succinate as carbon source was optimized and the process was bioaugmented with heterotrophic nitrification-aerobic denitrification bacteria for the treatment of real wastewater. The results showed that after bioaugmentation, the total nitrogen removal efficiency of the aeration SPD process increased by 50.46 % under condition of dissolved oxygen (DO) 3 mg/L. According to Illumina MiSeq sequencing and correlation analyses, the microbial community can perform SND under the conditions of DO 5 mg and HRT 6 h, but is susceptible to DO. Bioaugmentation mainly affected the carbon source metabolic network with heterotrophic bacteria Methyloversatilis, Thiothrix, and norank_Lentimicrobiaceae as nodes to change the community structure, thereby improving the performance of the functional microbial community. Kyoto Encyclopedia of Genes and Genomes analysis suggested that narB, narG, narH, nirK and narI were the key genes involved in the response to bioaugmentation. This work provides new insights for the application of the SPD process in wastewater treatment.

Response of wastewater treatment performance, microbial composition and functional genes to different C/N ratios and carrier types in MBBR inoculated with heterotrophic nitrification-aerobic denitrification bacteria.

To operate the moving bed biofilm reactor inoculated with HN-AD bacteria (B-MBBR) instead of activated sludge for livestock and poultry breeding wastewater (LPBW) disposal in most efficient manner, nitrogen removal (NR) efficiency and microbial composition of two MBBRs with different carrier types under various C/N ratios were explored. Results indicated that the performance on NR greatly various in different carrier types under various C/N ratios. Attributing to the bacterial protection provided by the porous structure of polyvinyl alcohol (PVA) gel, MBBR using PVA gel as the carrier exhibited a more stable NR performance (range from 78.05% to 83.76%) versus that using Kaldnes (K1) as the carrier (range from 78.05% to 83.76%). Besides, microbial analysis indicated that MBBR with PVA gel as the carrier is conducive to the growth of oligotrophic and HN-AD bacteria (Paracoccus and Acinetobacter), and the highest relative abundance was 16.37% at C/N ratio of 6.

Baicalein Protects Against Aspergillus fumigatus Keratitis by Reducing Fungal Load and Inhibiting TSLP-Induced Inflammatory Response.

Purpose: To investigate the antifungal and anti-inflammatory effects of baicalein on Aspergillus fumigatus (A. fumigatus) keratitis and the underlying mechanisms. Methods: The noncytotoxic antifungal concentration of baicalein was determined using CCK8, cell scratch assay, minimum inhibitory concentration, biofilm formation, scanning electron microscopy, propidium iodide uptake test and adherence assay in vitro and Draize test in vivo. In fungal keratitis (FK) mouse models, clinical score and plate count were used to evaluate FK severity, and myeloperoxidase assay and immunofluorescence staining were performed to examine neutrophil infiltration and activity. Real-time PCR, ELISA, and Western blot were performed to explore the anti-inflammatory activity of baicalein and the underlying mechanisms in vivo and in vitro. Results: Baicalein at 0.25 mM (noncytotoxic) significantly inhibited A. fumigatus growth, biofilm formation, and adhesion in vitro. In A. fumigatus keratitis mice, baicalein mitigated FK severity, reduced fungal load, and inhibited neutrophil infiltration and activity. Baicalein not only suppressed mRNA and protein levels of proinflammatory factors IL-1ß, IL-6, and TNF-α, but also inhibited the expression of thymic stromal lymphopoietin (TSLP) and TSLP receptor (TSLPR) in vivo and in vitro. In HCECs, mRNA and protein levels of IL-1ß, IL-6, and TNF-α were significantly lower in the TSLP siRNA-treated group, while higher in the rTSLP-treated group than in the corresponding control. Baicalein treatment significantly inhibited rTSLP induced the expression of IL-1ß, IL-6, and TNF-α. Conclusions: Baicalein plays a protective role in mouse A. fumigatus keratitis by inhibiting fungal growth, biofilm formation, and adhesion, and suppressing inflammatory response via downregulation of the TSLP/TSLPR pathway.

Multifunctional Polydopamine Particles as a Thermal Stability Modifier to Prepare Antifouling Melt Blend Composite Membranes.

This study reports a novel, multifunctional, and easily obtained modifier to support the rapid advancements in the field of filtration. Polydopamine (PDA) particles (PDAPs) have been reported as a filler for constructing polymer composites, but because of their poor thermal stability, the use of PDAPs in high-temperature blend melt systems to construct antifouling membranes was rare. In this paper, high-thermal-stability methoxy polyethylene glycol amine (mPEG-NH2)-functionalized PDA nanoparticles (mPDAPs) were first used as a modifier in high-temperature blend melt polymer composites to construct antifouling composite membranes. First, high-thermal-stability mPDAPs with an average diameter of about 390 nm were prepared by immobilized mPEG-NH2 on the PDAP surface, then melt blend mPDAPs with ultrahigh-molecular-weight polyethylene/liquid paraffin (LP) solution and thermally reduced phase separation (TIPS) to construct antifouling membranes. A combination of properties including mechanical properties, filtration efficiency, and antifouling properties of hybrid composite membranes was investigated and demonstrated that mPDAPs were an efficient modifier for high-temperature melt blending systems. The aim of this study was to provide an effective approach to improve the membrane filtration performance by bulk hybrid modification of multifunctional nanoparticles.

Honokiol Reduces Fungal Load, Toll-Like Receptor-2, and Inflammatory Cytokines in Aspergillus fumigatus Keratitis.

Purpose: We characterized the effects of Honokiol (HNK) on Aspergillus fumigatus-caused keratomycosis and the underlying mechanisms. HNK is known to have anti-inflammatory and antifungal properties, but the influence on fungal keratitis (FK) remains unknown. Methods: In ex vivo, minimum inhibitory concentration and Cell Count Kit-8 assay were carried out spectrophotometrically to provide preferred concentration applied in vivo. Time kill assay pointed that HNK was fungicidal and fungistatic chronologically. Adherence assay, crystal violet staining, and membrane permeability assay tested HNK effects on different fungal stages. In vivo, clinical scores reflected the improvement degree of keratitis outcome. Myeloperoxidase (MPO) assay, flow cytometry (FCM), and immunohistofluorescence staining (IFS) were done to evaluate neutrophil infiltration. Plate count detected HNK fungicidal potentiality. RT-PCR, Western blot, and enzyme-linked immunosorbent assay (ELISA) verified the anti-inflammatory activity of HNK collaboratively. Results: In vitro, MIC90 HNK was 8 µg/mL (no cytotoxicity), and Minimal Fungicidal Concentration (MFC) was 12 µg/mL for A. fumigatus. HNK played the fungistatic and fungicidal roles at 6 and 24 hours, respectively, inhibiting adherence at the beginning, diminishing biofilms formation, and increasing membrane permeability all the time. In vivo, HNK improved C57BL/6 mice outcome by reducing disease severity (clinical scores), neutrophil infiltration (MPO, FCM, and IFS), and fungal loading (plate count). RT-PCR, Western blot, and ELISA revealed that HNK downregulated mRNA and protein expression levels of Toll-like receptor-2 (TLR-2), high mobility group box 1 (HMGB1), IL-1ß, and TNF-α. Conclusions: Our study suggested HNK played antifungal and anti-inflammatory roles on keratomycosis by reducing survival of fungi, infiltration of leucocytes, and expression of HMGB1, TLR-2, and proinflammatory cytokines, providing a potential treatment for FK.