Development of an effective method for purifying trypsin using a recombinant inhibitor.

A trypsin affinity material was prepared by covalently immobilizing buckwheat trypsin inhibitor (BTI) on epichlorohydrin-activated cross-linked agarose gel (Selfinose CL 6 B). The optimal conditions for activating Selfinose CL 6 B were 15 % epichlorohydrin and 0.8 M NaOH at 40 °C for 2 h. The optimal pH for immobilizing BTI was 9.5. BTI-Sefinose CL 6 B showed a maximum adsorption capacity of 2.25 mg trypsin/(g support). The material also displayed good reusability, retaining over 90 % of its initial adsorption capacity after 30 cycles. High-purity trypsin was obtained from locust homogenate using BTI-Selfinose CL 6 B through one-step affinity chromatography. The molecular mass and Km value of locust trypsin were determined as 27 kDa and 0.241 mM using N-benzoyl-DL-arginine-nitroanilide as substrate. The optimal temperature and pH of trypsin activity were 55 °C and 9.0, respectively. The enzyme exhibited good stability in the temperature range of 30-50 °C and pH range of 4.0-10.0. BTI-Selfinose CL 6 B demonstrates potential application in the preparation of high-purity trypsin and the discovery of more novel trypsin from various species.

Improved Alkaline Hydrogen Evolution Performance of Dealloying Fe75-xCoxSi12.5B12.5 Electrocatalyst.

The electrocatalytic performance of a Fe65Co10Si12.5B12.5 Fe-based compounds toward alkaline hydrogen evolution reaction (HER) is enhanced by dealloying. The dealloying process produced a large number of nanosheets on the surface of NS-Fe65Co10Si12.5B12.5, which greatly increased the specific surface area of the electrode. When the dealloying time is 3 h, the overpotential of NS-Fe65Co10Si12.5B12.5 is only 175.1 mV at 1.0 M KOH and 10 mA cm-2, while under the same conditions, the overpotential of Fe65Co10Si12.5B12.5 is 215 mV, which is reduced. In addition, dealloying treated electrodes also show better HER performance than un-dealloying treated electrodes. With the increase in Co doping amount, the overpotential of the hydrogen evolution reaction decreases, and the hydrogen evolution activity is the best when the addition amount of Co is 10%. This work not only provides a basic understanding of the relationship between surface activity and the dealloying of HER catalysts, but also paves a new way for doping transition metal elements in Fe-based electrocatalysts working in alkaline media.

Interaction of diesel exhaust particulate matter with mucins in simulated saliva fluids: Bioaccessibility of heavy metals and potential health risks.

Air pollution is one of the major environmental risks threatening human health, diesel exhaust particulate matter (DEPM) is an important source of urban air pollution, and oral ingestion is the primary route of exposure to atmospheric particulate matter. This study examined the bioaccessibility of Cr, Fe, and Zn in DEPM within simulated saliva fluids through in vitro experiments, interactions between the particles and mucins, and the mechanisms underlying the oxidative damage they cause. The results indicated that the interaction between DEPM and mucins altered the dispersibility, surface charge, and wettability of the particles, leading to increased release of heavy metals. Protein adsorption experiments and characterizations revealed that the adsorption of mucin by the particles resulted in a complexation reaction between the metals in the DEPM and the mucins, accompanied by fluorescence quenching of the protein. In addition, free radical assays and correlation analyses revealed that environmentally persistent free radicals generated by DEPM induce the production of reactive oxygen species (O2·-, HOOH, and·OH), which damage the secondary structure of mucins and increase the risk of oral diseases. Our study is the first to reveal the interaction between DEPM and mucins in saliva, elucidating the mechanisms of DEPM-induced oxidative damage. This is significant for understanding the oral health risks posed by the ingestion of atmospheric particulate matter.

Huaier-induced suppression of cancer-associated fibroblasts confers immunotherapeutic sensitivity in triple-negative breast cancer.

BACKGROUND: Triple-negative breast cancer (TNBC) is the most intractable subgroup of breast neoplasms due to its aggressive nature. In recent years, immune checkpoint inhibitors (ICIs) have exhibited potential efficacy in TNBC treatment. However, only a limited fraction of patients benefit from ICI therapy, primarily because of the suppressive tumor immune microenvironment (TIME). Trametes robiniophila Murr (Huaier) is a traditional Chinese medicine (TCM) with potential immunoregulatory functions. However, the underlying mechanism remains unclear. PURPOSE: The present study aimed to investigate the therapeutic role of Huaier in the TIME of TNBC patients. METHODS: Single-cell RNA sequencing (scRNA-seq) was used to systematically analyze the influence of Huaier on the TNBC microenvironment for the first time. The mechanisms of the Huaier-induced suppression of cancer-associated fibroblasts (CAFs) were assessed via real-time quantitative polymerase chain reaction (qRT‒PCR) and western blotting. A tumor-bearing mouse model was established to verify the effects of the oral administration of Huaier on immune infiltration. RESULTS: Unsupervised clustering of the transcriptional profiles suggested an increase in the number of apoptotic cancer cells in the Huaier group. Treatment with Huaier induced immunological alterations from a "cold" to a "hot" state, which was accompanied by phenotypic changes in CAFs. Mechanistic analysis revealed that Huaier considerably attenuated the formation of myofibroblastic CAFs (myoCAFs) by impairing transforming growth factor-beta (TGF-ß)/SMAD signaling. In mouse xenograft models, Huaier dramatically modulated CAF differentiation, thus synergizing with the programmed cell death 1 (PD1) blockade to impede tumor progression. CONCLUSIONS: Our findings demonstrate that Huaier regulates cancer immunity in TNBC by suppressing the transition of CAFs to myoCAFs and emphasize the crucial role of Huaier as an effective adjuvant agent in immunotherapy.

Role of serotonin neurons in the dorsal raphe nucleus in heroin self-administration and punishment.

One hallmark of substance use disorder is continued drug use despite negative consequences. When drug-taking behavior is punished with aversive stimuli, i.e. footshock, rats can also be categorized into punishment-resistant or compulsive vs. punishment-sensitive or non-compulsive phenotypes. The serotonin (5-hydroxytryptamine, 5-HT) system modulates responses to both reward and punishment. The goal of the current study was to examine punishment phenotypes in heroin self-administration and to determine the role of dorsal raphe nucleus (DRN) 5-HT neurons in both basal and punished heroin self-administration. First, rats were exposed to punished heroin self-administration and neuronal excitability of DRN 5-HT neurons was compared between punishment-resistant and punishment-sensitive phenotypes using ex vivo electrophysiology. Second, DRN 5-HT neuronal activity was manipulated in vivo during basal and punished heroin self-administration using chemogenetic tools in a Tph2-iCre rat line. While rats separated into punishment-resistant and punishment-sensitive phenotypes for punished heroin self-administration, DRN 5-HT neuronal excitability did not differ between the phenotypes. While chemogenetic inhibition of DRN 5-HT neurons was without effect, chemogenetic activation of DRN 5-HT neurons increased both basal and punished heroin self-administration selectively in punishment-resistant animals. Additionally, the responsiveness to chemogenetic activation of DRN 5-HT neurons in basal self-administration and motivation for heroin in progressive ratio each predicted resistance to punishment. Therefore, our data support the role for the DRN 5-HT system in compulsive heroin self-administration.

The impact of economic agglomeration on China's urban public health.

Introduction: This study aims to explore the impact of economic agglomeration on the urban prosperity through economies of scale and agglomeration, it may also affect the public health of the agglomeration area. Methods: This paper takes 280 cities in China as the research object, and explores the impact of economic agglomeration on public health through a two-way fixed effects model, instrumental variable method, and generalized moment estimation. Results: The results indicate that: (1) the improvement of China's economic agglomeration can significantly promote urban public health, and economic agglomeration is a prerequisite for the improvement of urban public health, but there is no reverse causal relationship. (2) The enhancement of economic agglomeration in Northeast China has the greatest promotion effect on public health, followed by the eastern, western, and central regions; The economic agglomeration enhancement of the pilot medical group in closely connected cities has a greater promoting effect on public health than the pilot medical group in non-closely connected cities. (3) Empirical results based on micro sample data show that the improvement of economic agglomeration will also promote the increase of the number of public hospitals in cities. Discussion: This study emphasizes the important role of economic accumulation in the improvement of urban public health and provides empirical support for future economic development policies and practices.

Long-lived lung megakaryocytes contribute to platelet recovery in thrombocytopenia models.

Lung megakaryocytes (Mks) are largely extravascular with an immune phenotype (1). Because bone marrow (BM) Mks are short-lived it has been assumed that extravascular lung Mks are constantly 'seeded' from the BM. To investigate lung Mk origins and how that impacts their functions, we developed methods to specifically label lung Mks using CFSE dye and biotin delivered oropharyngeal. Labeled lung Mks were present for up to four months, while BM Mks had a <1 week lifespan. In a parabiosis model, lung Mks were partially replaced over 1-month from a circulating source. Unlike tissue-resident macrophages, using MDS1-Cre-ERT2 TdTomato mice, we found that lung Mks arise from hematopoietic stem cells. However, studies with FlkSwitch mTmG mice showed that lung Mks are derived from a Flt3-independent lineage that does not go through a multipotent progenitor. CFSE labeling to track lung Mk-derived platelets showed that about 10% of circulating platelets are lung resident Mk-derived at steady state, but in sterile thrombocytopenia this was doubled (about 20%). Lung-derived platelets were similarly increased in a malaria infection model (Plasmodium yoelii) typified by thrombocytopenia. These studies indicate that lung Mks arise from a Flt3-negative BM source, are long-lived, and contribute more platelets during thrombocytopenia.

Submicroscopic magnetite may be ubiquitous in the lunar regolith of the high-Ti region.

Magnetite is rare on the Moon. The ubiquitous presence of magnetite in lunar soil has been hypothesized in previous Apollo Mössbauer spectroscopy and electron spin resonance studies, but there is currently no mineralogical evidence to prove it. Here, we report a large number of submicroscopic magnetite particles embedded within iron-sulfide on the surface of Chang'e-5 glass, with a close positive correlation between magnetite content and the TiO2 content of the surrounding glass. The morphology and mineralogy of the iron-sulfide grains suggest that these magnetite particles formed via an impact process between iron-sulfide droplets and silicate glass melt, and ilmenite is necessary for magnetite formation. Magnetite in lunar glass is a potential candidate for the "magnetite-like" phase detected in the Apollo era and suggests that impact-induced submicroscopic magnetite may be ubiquitous in high-Ti regions of the Moon. Moreover, these impact-induced magnetite particles may be crucial for understanding the lunar magnetic anomalies and mineral components of the deep Moon.

Enzymatic oxidation increases the antibacterial activity of myricetin against Staphylococcus aureus.

Myricetin (MYR) is a flavonoid with favorable biological activities. In this study, MYR oxidation products (MYRox) were generated through enzymatic oxidation of MYR using horseradish peroxidase. The results showed enzymatic oxidation enhanced the water solubility and antibacterial activity against Staphylococcus aureus (S. aureus) of MYR. Further experiments showed the antibacterial effects of MYRox were conferred by MYR organic phase oxidation products (MYRoo). Both MYR and MYRoo could disrupt the cell membrane integrity, bind to the genomic DNA, affect protein synthesis and degradation, and alter the ROS levels in S. aureus. However, they exerted these effects with different strengths and ways. Finally, MYR or MYRoo can be used as an inhibitor against S. aureus in the cabbage food system, with MYRoo having better effect. This study demonstrated that enzymatic oxidation is an effective approach to improve the water solubility and antibacterial activity of MYR, enhancing its potential application in food preservation.

Production of protein-epigallocatechin gallate conjugates using free radicals induced by ultrasound and their gelation behavior.

In this study, free radicals generated by ultrasound were used to prepare conjugates of food proteins (soybean protein isolates, sodium caseinate and gelatin) with epigallocatechin gallate (EGCG). The changes in free amino and sulfhydryl group contents were used to confirm the occurrence of conjugation. The formation of covalent interactions on surface hydrophobicity, functional groups, structures, thermal stability, and gelation behavior of three proteins were investigated. The results showed that conjugation led to decrease in free amino and sulfhydryl group contents, reduction in the intensity of amide A and fluorescence intensity, and increase in ß-fold content. The conjugation also resulted in a decrease in surface hydrophobicity and thermal stability of soybean protein isolates and sodium caseinate, but an increase in the surface hydrophobicity and thermal stability of gelatin. Furthermore, the covalent bonding between proteins and EGCG improved gel strength, water holding capacity, and resulted in a denser and more compact microstructure.

The JNK-associated leucine zipper protein exerts a protective effect on renal parenchymal injury by limiting the inflammatory secretome in tubular cells.

JNK-associated leucine zipper protein (JLP) is a newly identified renal endogenous anti-fibrotic factor that is selectively enriched in renal tubular epithelial cells (TECs). The loss of JLP by TECs is a landmark event that heralds the progression of kidney fibrosis. JLP deficiency ensues a series of pathogenetic cellular processes that are conducive to fibrotic injury. Inflammatory injury is functionally relevant in fibrotic kidneys, and TECs play an essential role in fueling inflammation through aberrant secretions. It is speculated that the loss of JLP in TECs is associated with the relentless inflammation during the development of kidney fibrosis. This study examined the alteration of a panel of inflammatory signatures in TECs under kidney fibrotic circumstances using a Jlp gene-modified unilateral ureteral obstruction (UUO) mouse model or cultured HK-2 cells. It was found that a deficiency of JLP in TECs led to a significant increase in the secretion of inflammatory cytokines including interleukin-1ß (IL-1ß), tumor necrosis factor (TNF-α), and monocyte chemotactic protein-1 (MCP-1), overactivation of the nuclear factor (NF)-κB/c-Jun N-terminal kinase (JNK) pathway, as well as nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) inflammasome-mediated pyroptosis in response to pro-fibrotic damage. Additionally, the absence of JLP resulted in enhanced macrophage migration and fibroblast activation as paracrine effects elicited by injured TECs. In conclusion, the loss of JLP in TECs catalyses inflammatory injuries in the development of kidney fibrosis.

G protein-coupled receptor kinase 5 regulates thrombin signaling in platelets.

Background: Our prior genome-wide association study of thrombin-induced platelet aggregation identified a G protein-coupled receptor kinase 5 (GRK5) noncoding variant (rs10886430-G) that is strongly associated with increased platelet reactivity to thrombin. This variant predisposes to increased risk of stroke, pulmonary embolism, and venous thromboembolism. Objectives: To determine role of platelet specific GRK5 in platelet responses to agonists and injury. Methods: Platelets from GRK5 mutant mice have been shown to have increased thrombin sensitivity, indicating that GRK5 may be a negative regulator of platelet activation. However, this has not been studied in a platelet-specific manner. We therefore used platelet-specific GRK5 mutant mice and models of thrombosis and pulmonary embolism. Results: We now demonstrate that mice lacking GRK5 specifically in platelets had a mild increase in thrombin responses in vitro and a shortened time to arterial thrombosis in vivo. In addition, platelet GRK5 mutant mice had increased thrombin but not collagen-induced thrombus burden in a mouse model of pulmonary embolism. Conclusion: These data indicate that platelet GRK5 has a significant role in limiting platelet responses to thrombin.

Herpes Zoster Risk After Total Knee Replacement: a multicenter, propensity-score-matched cohort study in the United States.

Background: Total knee replacement (TKR) is a common surgical procedure for osteoarthritis (OA) patients. TKR may increase susceptibility to herpes zoster (HZ) by inducing immunosuppression, surgical stress, and nerve injury. However, limited data exist on the relationship between TKR and HZ. This study examined the risk of HZ over time among OA patients who underwent TKR and those who did not, using a large population-based cohort. Method: Utilizing the TriNetX research network, people with OA and underwent TKR were recruited as case group. After 1:1 propensity score matching, OA patients who never experienced TKR were included as control group. Covariates, including demographics, comorbidities, and laboratory data, were balanced using propensity score matching. A 5-year follow-up assessed the hazard ratio of incident HZ and related complications. Results: Compared to the control group, a significantly elevated risk of HZ was observed in the TKR cohort across 5-year follow-up period, with the hazard ratio of 1.223 (95% CI: 1.089-1.373). Zoster without complications presented 1.173-fold risk in TKR patients while comparing with non-TKR controls. However, most other secondary outcomes related to HZ complications-such as encephalitis, neurological involvement, ocular disease, and disseminated zoster-did not show a significant increase in risk. The risk of HZ was statistically significant for females and older adults in the TKR cohort than in the control cohort. Conclusions: OA patients who underwent TKR had an increased risk of HZ compared to those who did not receive the procedure, especially females and older adults. These findings highlight the need for HZ monitoring/prevention protocols and further research on mitigating viral reactivation after major joint surgery.

A colorimetric-aptamer-based assay for the determination of enrofloxacin through triggering the aggregation of gold nanoparticles.

Although enrofloxacin (ENR) is a widely used broad-spectrum antibiotic in veterinary medicine, its residues in animals can pose a risk to human health. Thus, we developed a new method for detecting ENR based on aptamers and AuNPs. In the absence of ENR, the aptamers attached to the surface of the AuNPs via electrostatic interactions to protect the AuNPs from NaCl, and the solution remained red. Conversely, the aptamer bonded with ENR, leading the aptamer to detach from the AuNP surface, and the color of the solution changed from red to blue. Based on this principle, ENR can be qualitatively detected by the naked eye and quantitatively detected by measuring the absorbance ratio at 650 nm and 530 nm. The experimental results showed a good linear relationship within the ENR concentration range of 0-400 nM, with a limit of detection (LOD) of 1.72 nM, which is satisfactory for detection in food safety. Additionally, this method has also been successfully applied to the detection of ENR in tap water, river water, milk, serum and urine, with good recovery rates and RSD values of less than 7%, indicating its great potential for ENR detection in environmental water samples. More importantly, the combination of this method with a smartphone platform provided great convenience for on-site and visual detection of ENR, offering promising applicability prospects.

Patients with hidradenitis suppurativa are associated with risk of new-onset glaucoma: a propensity-score-matched cohort study.

Background: Ocular comorbidities of hidradenitis suppurativa (HS) has been widely evaluated; however real-world evidence was scarce. Moreover, risk of glaucoma in HS patients remained unclear. This study aimed to evaluate the 5-year glaucoma risk in HS patients. Methods: This retrospective cohort study used the TriNetX database covering 2005-2017. In total, 53,281 HS patients were propensity score matched 1:1 to controls based on demographics, including comorbidities, medications, healthcare utilization, etc. Patients were followed for 5 years post-index date. Glaucoma risks were calculated based on hazard ratios and 95% confidence intervals (95% CI). Stratified analyses by sex and age were performed. Results: After matching, baseline characteristics were similar between groups. HS was associated with a 1.25 times higher 5-year glaucoma risk (95% CI, 1.10-1.42). The risk was significant within 1 year (HR=1.37; 95% CI, 1.03-1.82), 3 years (HR=1.31; 95% CI, 1.12-1.54), and 5 years post-index. In subgroup analysis, women had a 1.28 times higher risk (95% CI, 1.10-1.49). Patients aged 18-64 years (HR=1.33; 95% CI, 1.14-1.55) and ≥65 years (HR=1.33; 95% CI, 1.05-1.67) also presented elevated glaucoma risks. Conclusion: This real-world data analysis demonstrated a significantly increased 5-year glaucoma risk in HS patients versus matched controls. Ocular complications should be concerned while managing HS patients.

Boosting circularly polarized luminescence by optimizing off-centering octahedral distortion in zero-dimensional hybrid indium-antimony halides.

Chiral zero-dimensional hybrid metal halides (0D HMHs) are being extensively studied as they can directly generate circularly polarized luminescence (CPL) with high photoluminescence quantum yields (PLQYs), yet improving their luminescence dissymmetry factor (g lum) remains a challenge. This study proposes a general strategy to boost the g lum value of chiral 0D HMHs by optimizing the off-centering distortion of inorganic octahedra. Accordingly, (R/S-MBA)2(2MA)In0.95Sb0.05Cl6 (MBA = α-methylbenzylammonium, 2MA = dimethylamine) and (R/S-MBA)2(3MA)In0.95Sb0.05Cl6 (3MA = trimethylamine) with near-unity PLQYs are accordingly synthesized. With increasing the from 0.012 to 0.020, the |g lum| is accordingly increased from 7.8 × 10-3 to 2.0 × 10-2. Notably, the |g lum| can be further boosted to an impressive value of 3.8 × 10-2 while maintaining near-unity PLQYs by continuously increasing the . Experimental results reveal that the choice of achiral ligands and varied Sb3+ dopant concentrations can modulate the distribution and strength of hydrogen bonds around indium-antimony halogen octahedra, respectively, thus regulating the parameter of octahedra in 0D hybrid metal halides. Additionally, light-emitting diodes with a polarization of 1.6% are fabricated. This work sheds light on the relationship between the distortion of inorganic octahedra and the g lum value.

Clinical Efficacy of Transcatheter Arterial Chemoembolization Combined With Percutaneous Microwave Coagulation Therapy for Advanced Hepatocellular Carcinoma.

Background: The aim of the study was to explore the clinical efficacy of transcatheter arterial chemoembolization (TACE) combined with percutaneous microwave coagulation therapy (PMCT) for advanced hepatocellular carcinoma (HCC). Methods: Eighty-three advanced HCC patients were divided into the experimental group (TACE + PMCT, 57 cases) and the control group (TACE alone, 26 cases). They received TACE treatment first, and computed tomography (CT) or hepatic artery angiography was performed 3 - 4 weeks after each treatment. Based on the comprehensive evaluation of iodine oil deficiency, fistula recanalization, residual lesions, and lesion progression, TACE or PMCT treatment was selectively performed, and three consecutive treatments were considered as one treatment cycle. Results: The experimental group had a response rate (RR) of 49.1%, and the control group had a RR of 38.4%. The reduction rate of alpha-fetoprotein (AFP) in the experimental group was significantly higher than the control group (P < 0.05). The cumulative survival rates in the experimental at 1-, 1.5-, and 2-year post-treatment were higher than the control group. The cumulative recurrence and metastasis rates in the experimental at 1.5-, and 2-year post-treatment were significantly lower than those in the control group (P < 0.05). In addition, there were no significant differences in treatment-related complications in the two groups. Conclusions: The combined treatment of TACE and PMCT for advanced HCC is a safe, feasible, and effective treatment method, prolonging the survival time, and reducing the recurrence and metastasis rate, without increased toxic and side effects.

Feasibility analysis of continuous extraction of biomaterials from flocculant sludge and potential applications in the fire protection field.

The sludge contains many high-value biological materials. However, current extraction methods focus only on individual materials, neglecting the further extraction potential of the residual after extraction. This study used continuous extraction to extract extracellular polymeric substances (EPS) and proteins (PN) from sludge, verified the flame retardancy of EPS and the foaming properties of PN and finally analyzed the economic feasibility of continuous extraction. The results showed that continuous extraction increased the protein extraction from 857.11 mg/L to 1089.41 mg/L. EPS reduced the heat release rate of linen fabric from 379.2 (J/g·K) to 38.3 (J/g·K), and PN achieved foaming capacity and stability reaching 770% and 71%, meeting the standards of foam extinguishing agents. The binding form of EPS with linen fabric and the peptide content in PN are crucial factors affecting their application effectiveness. Economic analysis showed that continuous extraction reduced processing costs by 37.64% compared to traditional sludge disposal methods.