Optimization of 3D Printing Parameters for Alumina Ceramic Based on the Orthogonal Test.

In this paper, an alumina ink with good rheological properties was successfully prepared using pseudoboehmite as the main component, nitric acid as the colloidal solvent, and sesbania powder as the lubricant. The impact of nine different ink formulations, namely, Ink1 to Ink9, on the printability and rheological features of the ink was investigated. Consequently, it was found that Ink3 with 5 wt % nitric acid and 5 wt % sesbania powder exhibited the most favorable formability. This ink was utilized to fabricate alumina samples with direct ink writing (DIW) three-dimensional (3D) printing technology. The printed alumina samples were characterized using an automatic Brunauer-Emmett-Teller, X-ray diffractometer, Fourier transform infrared spectroscopy, and scanning electron microscope. To obtain the optimal printing parameters, a three-factor and three-level orthogonal test was designed to research the influences of different 3D printing parameters (filling ratio, nozzle diameter, and layer thickness) on the specific surface area, pore characteristics (size and volume), and radial crushing strength of the alumina specimens. The primary and secondary orders of the effects on the radial crushing strength and pore structure were determined through analysis of the experimental data. The experimental results showed that the alumina sample with a filling ratio of 80%, nozzle diameter of 0.6 mm, and layer thickness of 0.3 mm was found to have better strength of 48.07 ± 9.53 N/mm and specific surface area of 185.7315 m2/g. This study provides a theoretical base for the preparation of alumina carriers by DIW 3D printing.

In situ forming of PEG-NH2/dialdehyde starch Schiff-base hydrogels and their application in slow-release urea.

In this study, a biodegradable Schiff-base hydrogel urea, possessing substantial water retention and certain slow-release ability was designed and synthesized. Firstly, dialdehyde starch (DAS) and amine-terminated polyethylene glycol (PEG-(NH2)2) were synthesized using potato starch and polyethylene glycol. Then, a novel Schiff-base hydrogel (SH) was prepared through the in-situ reaction between the aldehyde group of DAS and the amino group of PEG-(NH2)2. Three SH based slow-release urea, designated as SHU1, SHU2, and SHU3 and distinguished by varying urea content, were obtained using SH as the substrate. Several characterizations and tests were conducted to determine the structure, thermal properties, morphology, swelling properties, sustainable use, water retention, and biodegradation properties of SH. Additionally, the slow-release behavior of SHU was studied. SEM results revealed that SH possessed a porous three-dimensional network structure, with a maximum water absorption capacity of 4440 % ± 6.23 %. Compared to pure urea, SHU exhibited better slow-release performance after 30 days of release in soil, with SHU1 having a residual nitrogen content of specifically 36.01 ± 0.57 % of the initial nitrogen content. A pot experiment with pakchoi substantiated the water retention and plant growth promotion properties of SHU. This study demonstrated a straightforward method for the preparation of starch-based Schiff-base hydrogels as fertilizer carriers.

Inhibition of Cell Apoptosis by Apicomplexan Protozoa-Host Interaction in the Early Stage of Infection.

Apicomplexan protozoa, which are a group of specialized intracellular parasitic protozoa, infect humans and other animals and cause a variety of diseases. The lack of research on the interaction mechanism between Apicomplexan protozoa and their hosts is a key factor restricting the development of new drugs and vaccines. In the early stages of infection, cell apoptosis is inhibited by Apicomplexan protozoa through their interaction with the host cells; thereby, the survival and reproduction of Apicomplexan protozoa in host cells is promoted. In this review, the key virulence proteins and pathways are introduced regarding the inhibition of cell apoptosis by the interaction between the protozoa and their host during the early stage of Apicomplexan protozoa infection. It provides a theoretical basis for the development of drugs or vaccines for protozoal diseases.

A highly selective ratio-metric fluorescent sensor for visualizing nitroreductase in hypoxic cells.

Herein, we have developed a novel single-molecular probe (NORP) for selective and accurate determination of NTR in living cells. It was discovered that up-regulation of endogenous NTR occurred in response to hypoxic stimulation, and there was a dependence between the NTR levels and the degree of hypoxia.

A study on rapid and stable catalytic reduction of 4-nitrophenol by 2-hydroxyethylamine stabilized Fe3O4@Pt and its kinetic factors.

The successful development of efficient and stable catalysts for 4-NP reduction reactions is beneficial to the environment and ecology. Fe3O4@Pt exhibits excellent catalytic performance for 4-NP reduction reaction due to the synergistic effect between Fe and Pt. But its structure and catalytic performance are extremely unstable. Here, we utilized the small-scale organic compound 2-hydroxyethylamine as surfactant to construct a stable composite nanomaterial. Then investigated the influence of monochromatic light (650 nm, 808 nm and 980 nm) and temperature on the kinetics of 4-NP reduction reaction by 2-hydroxyethylamine stabilized Fe3O4@Pt. The results indicate that both temperature and monochromatic light radiation can affect kinetic regulation. Increasing temperature can promote the catalytic rate, while monochromatic light radiation can induce agglomeration and inhibit the catalytic rate. This study opens up a new way for developing and regulating catalysts for heterogeneous catalysis reactions.

Carbon nanotube/Chitosan hydrogel for adsorption of acid red 73 in aqueous and soil environments.

Acid red 73 is an azo dye, and its residue can pollute the environment and seriously threaten human health and life. In this study, glutaraldehyde was used as the crosslinking agent, chitosan and polyvinyl alcohol were crosslinked under appropriate conditions to obtain a chitosan hydrogel film, and carbon nanotubes were dispersed in the chitosan hydrogel film. The FTIR, XRD, BET, SEM were applied to chatacterize the structure and the morphology of the absorbent and results showed that when the mass fraction of the carbon nanotubes was 1%, the structure was a three-dimensional network with microporous, and the water absorption reached to the maximum value of 266.07% and the elongation at break reached to a maximum of 98.87%. The ability to remove acid red 73 from aqueous and soil environments was evaluated by UV. In the aqueous samples, 70 mg of the adsorbent reached a saturated adsorption capacity of 101.07 mg/g and a removal rate of 92.23% at pH = 5. The thermodynamics conformed with the Langmuir adsorption isotherm and pseudo second-order adsorption kinetic models. In the soil samples, 100 mg of the adsorbent reached an adsorption capacity of 24.73 mg/g and removal rate of 49.45%. When the pH of the soil is between 4 and 7, the removal rate and adsorption capacity do not change much; hence, the pH should be maintained between 5.2 and 6.8, which is extremely suitable for the growth of general plants. Moreover, the experimental results demonstrated that the adsorbent maintained a good removal rate of acid red 73 over six adsorption cycles.

Repurposing the KCa3.1 Blocker Senicapoc for Ischemic Stroke.

Senicapoc, a small molecule inhibitor of the calcium-activated potassium channel KCa3.1, was safe and well-tolerated in clinical trials for sickle cell anemia. We previously reported proof-of-concept data suggesting that both pharmacological inhibition and genetic deletion of KCa3.1 reduces infarction and improves neurologic recovery in rodents by attenuating neuroinflammation. Here we evaluated the potential of repurposing senicapoc for ischemic stroke. In cultured microglia, senicapoc inhibited KCa3.1 currents with an IC50 of 7 nM, reduced Ca2+ signaling induced by the purinergic agonist ATP, suppressed expression of pro-inflammatory cytokines and enzymes (iNOS and COX-2), and prevented induction of the inflammasome component NLRP3. When transient middle cerebral artery occlusion (tMCAO, 60 min) was induced in male C57BL/6 J mice, twice daily administration of senicapoc at 10 and 40 mg/kg starting 12 h after reperfusion dose-dependently reduced infarct area determined by T2-weighted magnetic resonance imaging (MRI) and improved neurological deficit on day 8. Ultra-high-performance liquid chromatography/mass spectrometry analysis of total and free brain concentrations demonstrated sufficient KCa3.1 target engagement. Senicapoc treatment significantly reduced microglia/macrophage and T cell infiltration and activation and attenuated neuronal death. A different treatment paradigm with senicapoc started at 3 h and MRI on day 3 and day 8 revealed that senicapoc reduces secondary infarct growth and suppresses expression of inflammation markers, including T cell cytokines in the brain. Lastly, we demonstrated that senicapoc does not impair the proteolytic activity of tissue plasminogen activator (tPA) in vitro. We suggest that senicapoc could be repurposed as an adjunctive immunocytoprotective agent for combination with reperfusion therapy for ischemic stroke.

Ubiquitylome study reveals the regulatory effect of α-lipoic acid on ubiquitination of key proteins in tryptophan metabolism pathway of pig liver.

The decline in antioxidant defenses make it easily for human and animals to suffer from liver damage and diseases induced by oxidative stress, causing enormous losses to human health and livestock production. As one of the canonical protein post-translational modifications (PTMs), ubiquitination is widely involved in cell proliferation, apoptosis and damage/repair response, and is proven to be involved in the ability of mammals to resist oxidative stress. To explore whether α-lipoic acid (LA), a safe and efficient antioxidant, plays a role in regulating liver antioxidant status by PTMs, proteins in livers of pigs fed with LA were analyzed at the level of proteome and ubiquitylome. Based on proteome-wide enrichment of ubiquitination, a total of 7274 proteins were identified and 5326 were quantified, we also identified 1564 ubiquitination sites in 580 ubiquitinated proteins, among which there were 136 differentially ubiquitinated sites in 103 differentially ubiquitinated proteins upon LA. Further bioinformatics analysis showed that these differential proteins were mainly enriched in tryptophan metabolic pathway, and accompanied by significantly improvement of liver antioxidant capacity. We revealed the regulatory effect of LA on ubiquitination of kynurenine 3-monooxygenase (KMO) and other key proteins in tryptophan metabolism pathway of pig liver for the first time.

Hg(II) immobilization and detection using gel formation with tetra-(4-pyridylphenyl)ethylene and an aggregation-induced luminescence effect.

Tetra-(4-pyridylphenyl)ethylene (TPPE), featuring an aggregation-induced luminescence effect (AIE), has been synthesized and used for selective detection of Hg2+ in DMF/H2O (3:7, v/v) binary solutions. There was a color change from colorless to yellow in the detection of the Hg2+ ions, in addition to an increased fluorescence emission. This shows that TPPE will function as an excellent "turn-on" fluorescence probe in the detection Hg2+. Moreover, the interference of Al3+, Ba2+, Mn2+, Ca2+, Fe3+, Cu2+, Ag+, Cd2+, Co2+, Ni2+, Mg2+, Pb2+, Zn2+, and Cr3+ ions was found to be negligible under optimized solvent conditions. Cysteine and EDTA were also found to form TPPE-based fluorescent switches with the Hg2+ ions. The practical use of the TPPE sensor was also demonstrated by using a specific test kit. Characterization using FT-IR, NMR titration, UV titration, EDS, and HR-MS techniques showed that Hg2+ will form a 1:1 complex with TPPE. Also, the observation of a Tyndall effect, in addition to UV absorption and fluorescence spectra, did clearly demonstrate the presence of an AIE. More noteworthy, TPPE and Hg2+ were found to form a metal-organic gel (MOG) in the DMF solution. The SEM, TEM, ICP, and Zeta potential analyses confirmed that the fluorescent MOG could further adsorb an excess of Hg2+ ions. The BET analyses revealed that the MOG showed a type IV-H3 hysteresis loop according to the International Union of Pure and Applied Chemistry classification. The results of the XRD analysis and of the spectroscopic titrations show that a π-π stacking may be the auxiliary driving force for the gel formation, after that a coordination has taken place. These results indicate that further research on structurally simple metal ion fluorescent probes, which are based on the AIE, is promising for the achievement of a simultaneous fluorescent detection and adsorption of heavy metal pollutants.

Effects of voriconazole and fluconazole on the pharmacokinetics of almonertinib in rats by UPLC-MS/MS.

Almonertinib was included in the first-line treatment of non-small cell lung cancer with EGFR T790M mutations by the Chinese Society of Clinical Oncology in 2021. Considering that immunocompromised lung cancer patients are prone to opportunistic fungal infections, and most triazole antifungal drugs are moderate or strong inhibitors of CYP3A4, this study was conducted to develop and validate an accurate and rapid ultra-performance liquid chromatography tandem mass spectrometry method for quantifying almonertinib in plasma and for investigating the pharmacokinetic changes of almonertinib caused by voriconazole and fluconazole in rats. After liquid-liquid extraction with tert-butyl methyl ether, an XSelect HSS T3 column (2.1 × 100 mm, 2.5 µm, Waters) was used for the chromatographic separation of almonertinib and sorafenib-D3 (internal standard). The analytes were detected using an AB Sciex Triple Quad 5,500 mass spectrometer in the positive ionization mode. The method exhibited great linearity (0.5-200 ng/ml, r > 0.997) and stability under the established experimental conditions. All validation experiments were in accordance with the guidelines, and the results were all within the acceptable limits. This method was successfully applied to the researches of pharmacokinetics and drug interactions for almonertinib in rats. Voriconazole and fluconazole significantly altered the pharmacokinetic profiles of almonertinib and increased the systemic exposure of almonertinib in rats to different degrees, but further human trials should be conducted to validate the results.

Development of UPLC-MS/MS Method to Study the Pharmacokinetic Interaction between Sorafenib and Dapagliflozin in Rats.

Sorafenib (SOR), an inhibitor of multiple kinases, is a classic targeted drug for advanced hepatocellular carcinoma (HCC) which often coexists with type 2 diabetes mellitus (T2DM). Dapagliflozin (DAPA), a sodium-glucose cotransporter-2 inhibitor (SGLT2i), is widely used in patients with T2DM. Notably, co-administration of SOR with DAPA is common in clinical settings. Uridine diphosphate-glucuronosyltransferase family 1 member A9 (UGT1A9) is involved in the metabolism of SOR and dapagliflozin (DAPA), and SOR is the inhibitor of UGT1A1 and UGT1A9 (in vitro). Therefore, changes in UGT1A9 activity caused by SOR may lead to pharmacokinetic interactions between the two drugs. The objective of the current study was to develop an ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for the simultaneous determination of SOR and DAPA in plasma and to evaluate the effect of the co-administration of SOR and DAPA on their individual pharmacokinetic properties and the mechanism involved. The rats were divided into four groups: SOR (100 mg/kg) alone and co-administered with DAPA (1 mg/kg) for seven days, and DAPA (1 mg/kg) alone and co-administered with SOR (100 mg/kg) for seven days. Liquid-liquid extraction (LLE) was performed for plasma sample preparation, and the chromatographic separation was conducted on Waters XSelect HSS T3 column with a gradient elution of 0.1% formic acid and 5 mM ammonium acetate (Phase A) and acetonitrile (Phase B). The levels of Ugt1a7 messenger RNA (mRNA) were determined in rat liver and intestine using quantitative real-time polymerase chain reaction (qRT-PCR). The method was successfully applied to the study of pharmacokinetic interactions. DAPA caused a significant decrease in the maximum plasma concentrations (Cmax) and the area under the plasma concentration-time curves (AUC0-t) of SOR by 41.6% and 50.5%, respectively, while the apparent volume of distribution (Vz/F) and apparent clearance (CLz/F) significantly increased 2.85- and 1.98-fold, respectively. When co-administering DAPA with SOR, the AUC0-t and the elimination half-life (t1/2Z) of DAPA significantly increased 1.66- and 1.80-fold, respectively, whereas the CLz/F significantly decreased by 40%. Results from qRT-PCR showed that, compared with control, seven days of SOR pretreatment decreased Ugt1a7 expression in both liver and intestine tissue. In contrast, seven days of DAPA pretreatment decreased Ugt1a7 expression only in liver tissue. Therefore, pharmacokinetic interactions exist between long-term use of SOR with DAPA, and UGT1A9 may be the targets mediating the interaction. Active surveillance for the treatment outcomes and adverse reactions are required.

Effects of hydrogen peroxide and l-tryptophan on antioxidative potential, apoptosis, and mammalian target of rapamycin signaling in bovine intestinal epithelial cells.

Amino acids are primarily absorbed in the ruminant small intestine, and the small intestine is a target organ prone to oxidative stress, causing intestinal disfunction. Previous study suggested that l-Trp could benefit intestinal function and production performance. This study aimed to explore the effects of l-Trp on hydrogen peroxide (H2O2)-induced oxidative injury in bovine intestinal epithelial cells (BIEC) and the potential mechanism. The effects of l-Trp on cell apoptosis, antioxidative capacity, AA transporters, and the mammalian target of rapamycin (mTOR) signaling pathway were evaluated in BIEC treated with 0.8 mMl-Trp for 2 hours combined with or without H2O2 induction. In addition, to explore whether the effects of 0.8 mMl-Trp on oxidative stress were related to mTOR, an mTOR-specific inhibitor was used. The percentage of apoptosis was measured using flow cytometry. The relative gene abundance and protein expression in BIEC were determined using real-time PCR and Western blot assay, respectively. Results showed l-Trp at 0.4 and 0.8 mM enhanced the cell viability, and it was inhibited by l-Trp at 6.4 mM. l-Tryptophan at 0.4, 0.8, and 1.6 mM remarkably decreased the percentage of apoptosis and enhanced antioxidative capacity in H2O2-mediated BIEC. Moreover, l-Trp at 0.8 mM increased the relative gene abundance and protein expression of antioxidative enzymes and AA transporters, and the mTOR signaling pathway. The mTOR inhibitor lowered the protein expression of large neutral amino acid transporter 1, but the inhibition of mTOR did not alter the activities of catalase and superoxide dismutase or protein expression of alanine-serine-cysteine transporter 2 with or without H2O2 induction. l-Tryptophan increased catalase and superoxide dismutase activities in H2O2-mediated BIEC, although not with a present mTOR inhibitor. l-Tryptophan increased the protein expression of large neutral amino acid transporter 1 and alanine-serine-cysteine transporter 2 in H2O2-mediated BIEC with or without the presence of an mTOR inhibitor. The present work suggested that l-Trp supplementation could alleviate oxidative injury in BIEC by promoting antioxidative capacity and inhibiting apoptosis, and the mTOR signal played vital roles in the alleviation.

Pharmacokinetic Interactions between Canagliflozin and Sorafenib or Lenvatinib in Rats.

Hepatocellular carcinoma (HCC) and type 2 diabetes mellitus (T2DM) are common clinical conditions, and T2DM is an independent risk factor for HCC. Sorafenib and lenvatinib, two multi-targeted tyrosine kinase inhibitors, are first-line therapies for advanced HCC, while canagliflozin, a sodium-glucose co-transporter 2 inhibitor, is widely used in the treatment of T2DM. Here, we developed an ultra-performance liquid chromatography-tandem mass spectrometry method for the simultaneous determination of canagliflozin, sorafenib, and lenvatinib, and investigated the pharmacokinetic drug interactions between canagliflozin and sorafenib or lenvatinib in rats. The animals were randomly divided into five groups. Groups I-III were gavage administrated with sorafenib, lenvatinib, and canagliflozin, respectively. Group IV received sorafenib and canagliflozin; while Group V received lenvatinib and canagliflozin. The area under the plasma concentration-time curves (AUC) and maximum plasma concentrations (Cmax) of canagliflozin increased by 37.6% and 32.8%, respectively, while the apparent volume of distribution (Vz/F) and apparent clearance (CLz/F) of canagliflozin significantly decreased (30.6% and 28.6%, respectively) in the presence of sorafenib. Canagliflozin caused a significant increase in AUC and Cmax of lenvatinib by 28.9% and 36.2%, respectively, and a significant decrease in Vz/F and CLz/F of lenvatinib by 52.9% and 22.7%, respectively. In conclusion, drug interactions exist between canagliflozin and sorafenib or lenvatinib, and these findings provide a reference for the use of these drugs in patients with HCC and T2DM.

A self-ratiometric and selective electrochemical sensor for the detection of tyrosinase in mouse brain homogenate.

In this work, we developed a new organic electrochemical probe, 4-(prop-2-yn-1-yloxy)phenol, for the determination of tyrosinase. The designed probe contained a 4-hydroxybenzyloxy moiety for the specific determination of Tyr and a terminal alkyne group for chemical adsorption onto the electrode surface. The oxidation peak of the phenolic group from Pyyp decreased but increased for o-diphenol or o-quinone generated after the reaction between Tyr and Pyyp. The present sensor demonstrated a good linearity with Tyr activity in a dynamic range of 1.0-30 U mL-1. The limit of detection was as low as 0.28 ± 0.12 U mL-1. Moreover, this sensor demonstrated high selectivity for Tyr determination against metal ions, amino acids, ROS and neurotransmitters due to the specific recognition of Tyr. Finally, the developed sensor with high accuracy, high selectivity and long-term stability was successfully applied for evaluating Tyr activity in normal brain homogenate and brain homogenate with Parkinson's disease.

Trehalose Attenuates Oxidative Stress and Endoplasmic Reticulum Stress-Mediated Apoptosis in IPEC-J2 Cells Subjected to Heat Stress.

This study was carried out to investigate the effects of trehalose (Tre) on antioxidant capacity, endoplasmic reticulum stress (ERS) response and apoptosis of heat-stressed intestinal porcine epithelial cells (IPEC-J2). IPEC-J2 cells were cultured at 37 °C until the end of the experiment (control, CON); exposed to heat stress for 2 h (43 °C, HS); or pretreated with 0.1, 1, 5, 10, and 15 mM trehalose at 37 °C for 4 h prior to heat stress exposure for 2 h. The optimum level of trehalose for protecting against HS-induced cell injuries was determined to be 10 mM, as evidenced by the highest cellular viability and lowest malondialdehyde (MDA) content and lactate dehydrogenase (LDH) activity. Based on these, IPEC-J2 cells were divided into three groups: the first group was cultured at 37 °C until the end of the experiment (control, CON); the second group was exposed to heat stress for 2 h (43 °C, HS); the third group was pretreated with 10 mM trehalose for 4 h at 37 °C prior to heat stress exposure for 2 h (Tre + HS). The reactive oxygen species (ROS) content, superoxide dismutase (SOD) activity, mitochondrial membrane potential (MMP) changes, and expressions of the manganese superoxide dismutase (SOD2), ERS and apoptosis-related proteins were determined. Compared to the CON group, HS significantly increased ROS generation (p < 0.01), decreased SOD activity (p < 0.05), and downregulated protein expression of SOD2 (p < 0.01). Compared to the HS group, Tre supplementation reduced ROS levels and increased SOD activity and SOD2 expression to the levels that were comparable to the control (p < 0.05). The HS-induced ERS response was evidenced by the increased protein expressions of glucose-regulated protein 78 (GRP78) (p < 0.01), eukaryotic translation initiation factor 2α (p-eif2α) (p < 0.01), transcription activator 4 (ATF4) (p < 0.01), and the protein expression of C/EBP homologous protein (CHOP) (p < 0.01), which were the four hallmarks of ERS. The Tre + HS group showed lower expressions of GRP78 (p < 0.01), p-eif2α (p < 0.01), ATF4 (p < 0.01), and CHOP (p < 0.01) than that of the HS group. Tre pretreatment attenuated HS-induced mitochondrial apoptosis in IPEC-J2 cells, demonstrated by the increased MMP and decreased proapoptotic proteins active caspase 3, Bax, and cytochrome c (Cyt c). Taken together, trehalose can protect against HS-induced oxidative damage and endoplasmic reticulum stress-mediated apoptosis in IPEC-J2 cells. These data may provide a nutritional strategy for alleviating heat stress in pig production.

Determination of atypical antipsychotics in human plasma by UPLC-UV with polystyrene nanofibers as a solid-phase extraction sorbent.

A novel extraction procedure was developed using polystyrene (PS) nanofibers as a solid-phase extraction sorbent to collect atypical antipsychotics (AAPs) from human plasma. The extraction targets were then monitored by ultra high performance liquid chromatography with an ultraviolet detector system. Parameters affecting extraction efficiency such as fiber packing amount, wash solution, and eluted solvent were investigated. Under optimized conditions, the linear range of seven AAPs was 1-50 µgmL-1 (R 2 > 0.996). Inter-day and intra-day relative standard deviations were less than 15.1%, and relative error varied from -17.1% to 12.0%. Furthermore, 50.5-79.3% extraction recoveries were obtained. The lower limit of quantification was 1 µg mL-1, and detection limit was 0.5 µg mL-1. The method developed in this study may be applied to simultaneous quantification of seven AAPs in human plasma due to its simplicity, selectivity, and efficiency.

Study on the Mechanism of Bu-Shen-He-Mai Granules in Improving Renal Damage of Ageing Spontaneously Hypertensive Rats by Regulating Th17 Cell/Tregs Balance.

Methods: Blood pressure and urine biochemical indices were recorded. Renal blood flow was evaluated by renal ultrasonography. Transmission electron microscopy (TEM) and HE staining were used to assess kidney and spleen morphology. Renal fibrosis was assessed using Masson staining. Serum levels of IL-6, IL-10, and IL-17A were measured using ELISAs. The density of RORγ and Foxp3 in the spleen was observed by immunofluorescence staining. The levels of Th17 cells and Tregs in blood were detected via flow cytometry. Transcriptome sequencing was performed to screen the targets of BSHM granules in hypertensive kidneys. Results: BSHM granules decreased SBP by 21.2 mm·Hg and DBP by 8.8 mm·Hg in ageing SHRs (P < 0.05), decreased the levels of urine mALB, ß2-Mg, and NAG (P < 0.01), and improved renal blood flow and arteriosclerosis. BSHM granules increased IL-10 expression (P < 0.05) while decreasing IL-6 (P < 0.01) and IL-17A (P < 0.05) levels. BSHM granules improved Foxp3 density and the number of Tregs (P < 0.01) and reduced RORγt density and the number of Th17 cells (P < 0.01). Transcriptome sequencing identified 747 differentially expressed (DE) mRNAs in kidneys after BSHM treatment. GO analysis suggested that BSHM granules act through immunoregulation. Conclusions: BSHM granules attenuated hypertensive renal damage in ageing SHRs, by significantly increasing Tregs and decreasing Th17 cells.

Influence of schisantherin A on the pharmacokinetics of lenvatinib in rats and its potential mechanism.

Background: Lenvatinib (LEN) is approved as first-line therapy for advanced hepatocellular carcinoma (HCC). Schisantherin A (STA) can exert hepatoprotective and anti-tumor effects. The clinical combination of LEN and STA is very common, especially for patients with advanced HCC, but the effect of STA on the pharmacokinetics of LEN is unclear. This study aimed to investigate the effects of STA on the pharmacokinetics of LEN in rats and explore its potential mechanism. Methods: Male Sprague-Dawley (SD) rats were orally administered different doses of STA or vehicle control for 7 consecutive days, and 1.2 mg/kg of LEN was given on day 7. The messenger RNA (mRNA) and protein expression levels in the intestines and liver were investigated by reverse transcription quantitative polymerase chain reaction (RT-qPCR) and western blot. Results: It was revealed that STA increased the oral bioavailability of LEN. The area under the curve from time 0 to infinity (AUC0-∞) and maximum plasma concentration (Cmax) of LEN after co-administration with STA (20 mg/kg) increased by 54.3% (3,396.73±989.35 vs. 5,240.03±815.49 µg/L/h) and 54.8% (490.64±124.20 vs. 759.66±152.75 µg/L), respectively. The clearance decreased from 0.38±0.12 to 0.23±0.04 L/h/kg, and the apparent volume of distribution (Vz) decreased from 10.83±3.19 to 6.35±1.38 L/kg in the presence of 20 mg/kg STA. In addition, the expression of P-glycoprotein (P-gp) mRNA and protein in the intestines was markedly decreased. Conclusions: This study showed that STA increased the bioavailability of LEN, probably due to inhibition of P-gp in the intestine, thereby increasing systemic absorption of LEN. Thus, there is an interaction between the two drugs, and careful monitoring must be conducted when they are used in combination.

Leucine protects bovine intestinal epithelial cells from hydrogen peroxide-induced apoptosis by alleviating oxidative damage.

BACKGROUND: The present study aimed to investigate whether leucine (Leu) alleviates oxidative injury in bovine intestinal epithelial cells (BIECs) induced by hydrogen peroxide (H2 O2 ), as well as the underlying molecular mechanisms. RESULTS: BIECs were treated with H2 O2 (1 mmol L-1 ) and/or Leu (0, 0.9, 1.8 or 3.6 mmol L-1 ) for 2 h. Leu increased cell viability (P < 0.05) and decreased the release of lactate dehydrogenase (P < 0.05) in BIECs challenged by H2 O2 . Then, the cells were treated with H2 O2 (1 mmol L-1 ) and/or Leu (1.8 mmol L-1 ) for 2 h. Compared with the H2 O2 group, cells treated with Leu and Leu + H2 O2 exhibited increased (P < 0.05) mRNA and protein expression of superoxide dismutase 2 (SOD2), catalase (CAT), glutathione peroxidase 1 (GPx1), heme oxygenase 1 (HO-1) and nuclear factor erythroid 2-related factor 2 (Nrf2). BIECs treatment with Leu significantly reduced (P < 0.05) apoptosis induced by H2 O2 . BIECs were transfected with Nrf2 small interfering RNA (siRNA) for 48 h and/or treated with H2 O2 (1 mmol L-1 ) and/or Leu (1.8 mmol L-1 ) for another 2 h. Transfection with Nrf2 siRNA abrogated the protective effect of Leu against H2 O2 -induced apoptosis and the mRNA and protein expression of SOD2 (P < 0.05). CONCLUSION: These results indicate that Leu promotes the relative expression of antioxidant enzymes (SOD2, CAT and GPx1) and phase II detoxification enzymes (HO-1) by upregulating nuclear Nrf2 and activating the Nrf2 signaling pathway, thus enhancing the antioxidant capacity of cells. © 2022 Society of Chemical Industry.

A Simple UPLC/MS-MS Method for Simultaneous Determination of Lenvatinib and Telmisartan in Rat Plasma, and Its Application to Pharmacokinetic Drug-Drug Interaction Study.

Lenvatinib is a multi-targeted tyrosine kinase inhibitor that inhibits tumor angiogenesis, but hypertension is the most common adverse reaction. Telmisartan is an angiotensin receptor blocker used to treat hypertension. In this study, a simple ultra-performance liquid chromatography-tandem mass spectrometry method was developed for the simultaneous determination of lenvatinib and telmisartan, and it was applied to the pharmacokinetic drug interaction study. Plasma samples were treated with acetonitrile to precipitate protein. Water (containing 5 mM of ammonium acetate and 0.1% formic acid) and acetonitrile (0.1% formic acid) were used as the mobile phases to separate the analytes with gradient elution using a column XSelect HSS T3 (2.1 mm × 100 mm, 2.5 µm). Multiple reaction monitoring in the positive ion mode was used for quantification. The method was validated and the precision, accuracy, matrix effect, recovery, and stability of this method were reasonable. The determination of analytes was not interfered with by other substances in the blank plasma, and the calibration curves of lenvatinib and telmisartan were linear within the range of 0.2-1000 ng/mL and 0.1-500 ng/mL, respectively. The results indicate that lenvatinib decreased the systemic exposure of telmisartan. Potential drug interactions were observed between lenvatinib and telmisartan.