Discovery and Optimization of Potent, Efficacious and Selective Inhibitors Targeting EGFR Exon20 Insertion Mutations.

Herein, we report the identification and optimization of a series of potent inhibitors of EGFR Exon20 insertions with significant selectivity over wild-type EGFR. A strategically designed HTS campaign, multiple iterations of structure-based drug design (SBDD), and tactical linker replacement led to a potent and wild-type selective series of molecules and ultimately the discovery of 36. Compound 36 is a potent and selective inhibitor of EGFR Exon20 insertions and has demonstrated encouraging efficacy in NSCLC EGFR CRISPR-engineered H2073 xenografts that carry an SVD Exon20 insertion and reduced efficacy in a H2073 wild-type EGFR xenograft model compared to CLN-081 (5), indicating that 36 may have lower EGFR wild-type associated toxicity.

Exploring Salivary Iodine Concentration as a Biomarker for Iodine Status and Thyroid Nodules in Females From Different Water Iodine Areas: a Cross-sectional Study.

BACKGROUND: It is unclear whether salivary iodine concentration (SIC) can assess iodine status in females from different water iodine regions. OBJECTIVES: Through a cross-sectional study, we explored the feasibility of SIC as a biomarker to assess iodine status in females and develop optimal cutoff values. METHODS: A total of 1991 females were analyzed in this cross-sectional study from the coastal iodine-deficient areas (CIDAs), inland iodine-deficient areas (IIDAs), iodine-adequate areas (IAAs), iodine-excess areas (IEAs), and iodine extra-high areas (IEHAs). SIC, spot urine iodine concentration (SUIC), and daily total iodine intake (TII) were assessed, and ultrasonography was performed in all subjects. RESULTS: There was a positive correlation between SIC and SUIC (r = 0.67; 95% CI: 0.64, 0.69; P < 0.001), and TII (r = 0.47; 95% CI: 0.43, 0.50; P < 0.001). The prevalence of thyroid nodules (TN) showed an upward trend with SIC increasing (Z = -2.83; P-trend = 0.005). The area under the receiver-operating characteristic (ROC) curve for SIC to assess iodine deficiency was 0.62 (95% CI: 0.60, 0.65; P < 0.001) and 0.75 (95% CI: 0.73, 0.77; P < 0.001) for iodine excess. The cutoff values were as follows: SIC < 93.32 µg/L, iodine deficiency; 93.32-224.60 µg/L, iodine adequacy; and >224.60 µg/L, iodine excess. When SIC > 224.60 µg/L, the odds ratio (OR) for UIC > 300 µg/L, excessive TII, and the prevalence of TN were 6.44, 3.68, and 1.27 (95% CI: 4.98, 8.31; 2.83, 4.79; and 1.02, 1.56, respectively; P < 0.05); when SIC < 93.32 µg/L, the OR for UIC < 100 µg/L and insufficient TII were 2.34 and 1.94 (95% CI: 1.73, 3.14 and 1.33, 2.83, respectively; P < 0.05). CONCLUSIONS: Using SIC as a biomarker, females in CIDA exhibited mild iodine deficiency, those in IIDA and IAA demonstrated moderate iodine deficiency, and those in IEA and IEHA exhibited an excess of iodine, consistent with SUIC to assess iodine status. SIC can be used as a good biomarker to evaluate the iodine status in population.

Identifying chemical markers in wine-processed Salvia miltiorrhiza using ultrahigh-performance liquid chromatography-quadrupole-time-of-flight-tandem mass spectrometry.

To find the chemical markers of wine-processed Salvia miltiorrhiza (WSM), 76 constituents, including diterpenoid quinones and phenolic acids in Salvia miltiorrhiza (SM) and WSM, were profiled using ultrahigh-performance liquid chromatography-quadrupole-time-of-flight-tandem mass spectrometry (UPLC-Q-TOF-MS/MS) in positive- and the negative-ion modes. Thirty compounds were screened out as candidate differential components using chemometrics analysis, and the concentration of most compounds increased after processing with wine. Seven compounds, namely tanshinone IIA, magnesium lithospermate B, salvianolic acid G, cryptotanshinone, isocryptotanshinone, salvianolic acid B, and rosmarinic acid, were selected as chemical markers of WSM using variable importance of the project. This study revealed the chemical markers of WSM and confirmed that WSM can improve the extraction and solubility effect of chemical constituents.

Corn silk polysaccharides attenuate diabetic nephropathy through restoration of the gut microbial ecosystem and metabolic homeostasis.

Introduction: The pathogenesis of diabetic nephropathy (DN) is complex, inflammation is the central link among the inducing factors in the existing research, and the gutkidney axis could scientifically explain the reasons for the accumulation of chronic low-grade inflammation. As both a medicine and food, corn silk contains abundant polysaccharides. Historical studies and modern research have both confirmed its intervention effect on diabetes and DN, but the mechanism of action is unclear. Methods: In this study, a DN rat model was generated, and the therapeutic effect of corn silk polysaccharides (CSPs) was evaluated based on behavioral, histopathological and biochemical indicators. We attempted to fully understand the interactions between CSPs, the gut microbiota and the host at the systemic level from a gut microbiota metabolomics perspective to fundamentally elucidate the mechanisms of action that can be used to intervene in DN. Results: Research has found that the metabolic pathways with a strong correlation with CSPs were initially identified as glycerophosphate, fatty acid, bile acid, tyrosine, tryptophan and phenylalanine metabolism and involved Firmicutes, Bacteroides, Lachnospiraceae-NK4A136- group and Dubosiella, suggesting that the effect of CSPs on improving DN is related to changes in metabolite profiles and gut microbiota characteristics. Discussion: CSPs could be harnessed to treat the abnormal metabolism of endogenous substances such as bile acids and uremic toxins caused by changes in gut microbiota, thus alleviating kidney damage caused by inflammation. In view of its natural abundance, corn silk is safe and nontoxic and can be used for the prevention and treatment of diabetes and DN.

A Cross-Sectional Survey of Iodized Salt Usage in Dining Establishments - 13 PLADs, China, 2021-2022.

What is already known about this topic?: The National Iodine Deficiency Disease Surveillance system is exclusively focused on monitoring cooking salt used within households. Currently, there is a lack of nationally representative data on the use of iodized salt in dining establishments. What is added by this report?: This study evaluated 7,889 salt samples obtained from dining establishments located in 13 provincial-level administrative divisions across China. The findings indicated that coverage rate of iodized salt (CRIS) and the consumption rate of adequately iodized salt (CRAIS) were found to be 95.2% and 90.2%, respectively. Further, 880 samples were classified as iodized salt and 804 as adequately iodized salt. In coastal areas, the CRIS and CRAIS showed a significant decrease to 77.1% and 70.5%, respectively, when compared to the inland regions (P<0.01). What are the implications for public health practices?: The data compiled could potentially fill the void in the national data concerning the use of iodized salt in dining establishments throughout China. It is of the utmost importance to increase the awareness of restaurant operators, particularly those located in coastal areas, about the benefits of iodine supplementation. Moreover, they should be encouraged to use adequately iodized salt.

Survey of iodized salt coverage in areas with different water iodine concentrations and the iodine status of local women.

PURPOSE: There were only two definitions of iodine-deficient (water iodine concentration < 10 µg/L) and iodine-excess areas (water iodine concentration > 100 µg/L) in China before 2020. Areas with water iodine concentration between 10 and 100 µg/L implement the same policy as iodine-deficient areas to provide iodized salt. The definition of iodine-adequate areas was formulated in 2020 for the first time. The paper aims to investigate the coverage rate of iodized salt (CR) in different areas defined according to the latest national standards, evaluate the iodine status of local women, and provide a basis for the revision and improvement of relevant policies. METHODS: A total of 1948 women aged 18-60 were recruited from the iodine extra-high areas (IEHA), iodine-excess areas (IEA), iodine-adequate areas (IAA), inland iodine-deficient areas (IIDA), and coastal iodine-deficient areas (CIDA). Information on daily diet was collected with the Food Frequency Questionnaire. Drinking water, salt, food, and urine samples were collected and tested in our laboratory. Based on the recommended daily iodine intake, we assessed whether the subjects' daily iodine intake levels were adequate. RESULTS: The CR and the median urinary iodine concentrations (UICs) were 4.02% and 98.03 µg/L in CIDA, 89.74% and 144.93 µg/L in IIDA, 26.55% and 178.60 µg/L in IAA, 8.78% and 446.5 µg/L in IEA, 3.95% and 605.4 µg/L in IEHA, respectively. The differences among these five areas were statistically significant (P < 0.0001). The daily dietary iodine intakes were mainly from drinking water in IAA (63.92%), IEA (92.29%), and IEHA (92.93%), and were mainly from iodized salt in IIDA (59.22%) and food in CIDA (86.6%). CONCLUSION: Women in IAA and IIDA were in an adequate iodine state. Women in IEA and IEHA were in an iodine-excess state, and it is necessary to carry out water improvements projects. Women in CIDA were in a slight iodine-deficient state, and health education on scientific iodine fortification should be strengthened to increase iodine intake.

Effects of Broflanilide on Oxidative Stress and Expression of Apoptotic Genes in Zebrafish (Danio rerio) Gill.

Broflanilide exerted negative impacts on the gill of zebrafish. Thus, in this study, zebrafish gill was used to assess the apoptosis toxicity of broflanilide by determining the levels of reactive oxygen species (ROS), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and malondialdehyde (MDA) and apoptosis-related genes. The results found that the minimum threshold for the content and time of broflanilide affecting enzyme content and gene expression was 0.26 mg/L after 24 h exposure. After 96 h exposure, broflanilide could cause apoptosis and exerted significantly increased contents of ROS and MDA, while inhibiting the activities of SOD, CAT, and GPx at 0.26 and 0.57 mg/L. Broflanilide also had adverse effects on apoptosis-related genes, such as tumor protein p53 (p53), associated × (Bax), B-cell lymphama-2 (Bcl-2), caspase-3, caspase-9, and apoptotic protease activating factor-1(apaf-1), at 0.26 mg/L and 0.57 mg/L after 96 h exposure, respectively. These results provide new insight into the potential toxicity mechanisms of broflanilide in zebrafish gills.

Re-exploration of dietary iodine intake in Chinese adult males using a modified iodine balance study.

There is still controversy about optimal dietary iodine intake as the Universal Salt Iodization policy enforcement in China. A modified iodine balance study was thus conducted to explore the suitable iodine intake in Chinese adult males using the iodine overflow hypothesis. In this study, thirty-eight apparently healthy males (19·1 (sd 0·6) years) were recruited and provided with designed diets. After the 14-d iodine depletion, daily iodine intake gradually increased in the 30-d iodine supplementation, consisting of six stages and each of 5 d. All foods and excreta (urine, faeces) were collected to examine daily iodine intake, iodine excretion and the changes of iodine increment in relation to those values at stage 1. The dose-response associations of iodine intake increment with excretion increment were fitted by the mixed effects models, as well as with retention increment. Daily iodine intake and excretion were 16·3 and 54·3 µg/d at stage 1, and iodine intake increment increased from 11·2 µg/d at stage 2 to 118·0 µg/d at stage 6, while excretion increment elevated from 21·5 to 95·0 µg/d. A zero iodine balance was dynamically achieved as 48·0 µg/d of iodine intake. The estimated average requirement and recommended nutrient intake were severally 48·0 and 67·2 µg/d, which could be corresponded to a daily iodine intake of 0·74 and 1·04 µg/kg per d. The results of our study indicate that roughly half of current iodine intakes recommendation could be enough in Chinese adult males, which would be beneficial for the revision of dietary reference intakes.

Comparison of clinical outcomes between aggressive and non-aggressive intravenous hydration for acute pancreatitis: a systematic review and meta-analysis.

BACKGROUND: Current practice guidelines for optimal infusion rates during early intravenous hydration in patients with acute pancreatitis (AP) remain inconsistent. This systematic review and meta-analysis aimed to compare treatment outcomes between aggressive and non-aggressive intravenous hydration in severe and non-severe AP. METHODS: This study followed the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. We systematically searched PubMed, Embase and Cochrane Library for randomized controlled trials (RCTs) on November 23, 2022, and hand-searched the reference lists of included RCTs, relevant review articles and clinical guidelines. We included RCTs that compared clinical outcomes from aggressive and non-aggressive intravenous hydration in AP. Meta-analysis was performed using a random-effects model for participants with severe AP and non-severe AP. Our primary outcome was all-cause mortality, and several secondary outcomes included fluid-related complications, clinical improvement and APACHE II scores within 48 h. RESULTS: We included a total of 9 RCTs with 953 participants. The meta-analysis indicated that, compared to non-aggressive intravenous hydration, aggressive intravenous hydration significantly increased mortality risk in severe AP (pooled RR: 2.45, 95% CI: 1.37, 4.40), while the result in non-severe AP was inconclusive (pooled RR: 2.26, 95% CI: 0.54, 9.44). However, aggressive intravenous hydration significantly increased fluid-related complication risk in both severe (pooled RR: 2.22, 95% CI 1.36, 3.63) and non-severe AP (pooled RR: 3.25, 95% CI: 1.53, 6.93). The meta-analysis indicated worse APACHE II scores (pooled mean difference: 3.31, 95% CI: 1.79, 4.84) in severe AP, and no increased likelihood of clinical improvement (pooled RR:1.20, 95% CI: 0.63, 2.29) in non-severe AP. Sensitivity analyses including only RCTs with goal-directed fluid therapy after initial fluid resuscitation therapy yielded consistent results. CONCLUSIONS: Aggressive intravenous hydration increased the mortality risk in severe AP, and fluid-related complication risk in both severe and non-severe AP. More conservative intravenous fluid resuscitation protocols for AP are suggested.

Re-exploring the requirement of dietary iodine intake in Chinese female adults based on 'iodine overflow theory'.

PURPOSE: We re-explored the basal iodine requirement based on healthy Chinese female and a new iodine overflow theory was proposed for iodine balance study. METHODS: Thirty-six Chinese healthy female adults (age 20.7 ± 1.1) were recruited for this study, which included 40 days low iodine depletion period and six stages of 30 days supplementation period. Uniform diets with low iodine were provided and the content of iodine in the diet was regulated by dairy products. The total iodine intake from food and the total iodine excretion through 24-h urine and staged feces were completely gathered and monitored. The incremental (Δ) intake and excretion over the range were calculated. RESULTS: The iodine intake and excretion were 13.6 µg/day and 48.6 µg/day at the first stage, respectively. The incremental iodine intakes and excretions were 21.1 µg/day to 120.3 µg/day and 25.8 µg/day to 105.4 µg/day for the supplementation stages, respectively. According to the 'iodine overflow theory', the zero iodine balance (Δ iodine intake = Δ iodine excretion) derived from a mixed effect model indicated a mean iodine intake of 52.2 µg/d (1.0 µg/d kg). The RNI for iodine to healthy Chinese female adult was 73.1 µg/d (1.4 µg/d kg). CONCLUSION: A daily iodine intake of 52.2 µg/d may meet the basal iodine requirement for healthy Chinese female adults, and Chinese female may need more than 20% iodine intake than male based on the 'iodine overflow theory'. The trial was registered at the Chinese Clinical Trial Registry in May 2018 (No: ChiCTR1800016184).

Co-winemaking with Vitis amurensis Rupr. "Beibinghong" enhances the quality of Vitis vinifera L. cv. Cabernet Gernischt wine.

In some wine regions of China, Cabernet Gernischt (CG; Vitis vinifera L.) grape berries usually exhibit low pigment content and titratable acidity, and low sensory quality of the resulting wine. The aim of this study was to evaluate co-winemaking of CG wines using the red grape cultivar Beibinghong (BBH; Vitis amurensis Rupr.) at different proportions in terms of alcohols, phenolic compounds, and sensory properties of the wines. The results showed that the co-winemaking wines contained a similar content of higher alcohols, whereas the methanol content increased with an increase in BBH proportion, although this was still corresponded with the national standard. Significantly higher levels of titratable acidity were observed in co-winemaking wines at the ratio of 6:4 and 5:5, compared with monocultivar CG wines. All co-winemaking wines, except CG:BBH (9:1) wine, showed significantly higher levels of total anthocyanins, total phenolics, total tannins, and total flavan-3-ols. Further, individual phenolics, primarily diglucoside anthocyanins and non-anthocyanins (trans-ferulic acid, myricetin, viniferin, trans-caffeic acid, 3,4-dihydroxybenzoic acid), as important contributors to wine color intensity, permitted the differentiation of the wines via principal component analysis. In most cases, co-winemaking wines exhibited higher scores of the 10 sensory attributes on color, aroma, mouthfeel, and overall quality compared with monocultivar wines. Co-winemaking CG wines with BBH at 7:3 ratio demonstrated the highest scores of color intensity, aroma intensity, aroma quality, and overall quality. The results indicate that co-winemaking with V. amurensis grape variety may be useful to enhance V. vinifera wine quality by modifying wine composition. PRACTICAL APPLICATION: Cabernet Gernischt (CG) is the predominant grape cultivar used to prepare premium-quality wine in China; however, in some wine regions, CG wines have low levels of pigment and titratable acidity, and low sensory quality. Co-winemaking with another native grape cultivar, Beibinghong (BBH), which is characterized by a higher content of anthocyanins and acidity, provided sufficient experimental evidence of adjustments in the Vitis vinifera wine composition leading to improved wine sensory quality.

Neuroprotective Alkamides from the Aerial Parts of Achillea alpina L.

Three new alkamides, achilleamide B-D (1-3) along with five known alkamides (4-8) were isolated from the aerial parts of Achillea alpina L. Structures were elucidated by spectroscopic analysis. Modified Mosher's method and electronic circular dichroism (ECD) calculations were introduced for the absolute configuration of 3. The neuroprotective effects of all the compounds were evaluated by 6-hydroxydopamine (6-OHDA)-induced cell death in human neuroblastoma SH-SY5Y cells, with concentration for 50 % of maximal effect (EC50 ) values of 3.16-24.75 µM, and the structure-activity relationship was conducted.

Acute toxicity of broflanilide on neurosecretory system and locomotory behavior of zebrafish (Danio rerio).

Broflanilide, a novel meta-diamide insecticide, possesses moderate acute toxicity to zebrafish, with a 96-h median lethal concentration (96-LC50) of 0.76 mg/L. However, its effect on fish behavior and the underlying mechanisms are still unclear. The present study evaluated the effects of broflanilide on the zebrafish brain over a 96-h exposure by comparing the histopathological changes and relative expression of targeted genes with the behavioral metrics. The results of the toxicity test showed that broflanilide could cause deformities, such as deformation of the operculum and spinal curvature, at 0.6, 0.82 and 1.15 mg/L. Results also showed tissue damage and apoptosis in the cerebellum under 0.27 and 0.6 mg/L exposure. Additionally, broflanilide affected the neurotransmitters, metabolites and transcripts of genes associated with dopamine, gamma-aminobutyric acid expression. and the signaling pathways in zebrafish brains at 0.60 mg/L after 1 h and 96 h of exposure, while the levels of glutamate, glutamate decarboxylase, GABA transaminase, nicotinamide adenine dinucleotide (NADH) and adenosine triphosphate (ATP) were also inhibited at 0.27 mg/L after 96 h of exposure. The accumulated swimming distance was significantly longer and the average speed was significantly faster than the control at 0.27 and 0.6 mg/L after 1-h of exposure, while these metrics were lowered at 0.6 mg/L after 96 h of exposure. The study results demonstrates that broflanilide affects the zebrafish brain, neurotransmitters and associated fish behaviors. This study also provides deeper insight into the mechanistic understanding of the effects of broflanilide on the zebrafish brain.

Novel Step Floating Islands VDMOS with Low Specific on-Resistance by TCAD Simulation.

A novel VDMOS with Step Floating Islands VDMOS (S-FLI VDMOS) is proposed for the first time in this letter, in order to optimize the breakdown voltage (BV) and the specific on-resistance (Ron,sp). The innovative terminal technology of Breakdown Point Transfer (BPT) is applied to S-FLI VDMOS, which transfers the breakdown point from the high electric field region to the low electric field region, and the S-FLI VDMOS structure uses multiple layers of charge compensation blocks to generate multiple electric field peaks in the drift region in order to optimize the electric field distribution. In the TCAD simulation, the BV of the proposed S-FLI VDMOS is improved to 326 V, which is higher than that of 281 V for the conventional Si VDMOS with the same drift region length of 15 µm, and the Ron,sp is reduced from 21.54 mΩ·cm2 for the conventional Si VDMOS to 7.77 mΩ·cm2 for the S-FLI VDMOS. Compared with the conventional Si VDMOS, the current density of the effective current conduction path is increased when the forward bias is applied to the proposed device.

Antioxidant, antifungal, and aphicidal activity of the triterpenoids spinasterol and 22,23-dihydrospinasterol from leaves of Citrullus colocynthis L.

Terpenoids from natural plant sources are valuable for their diverse biological activities that have important roles in the medical and agrochemical industries. In this study, we assessed the antioxidant, antifungal, and aphicidal activities of a mixture of spinasterol and 22,23-dihydrospinasterol from the leaves of Citrullus colocynthis. We used 1,1-diphenyl-2-picrylhydrazyl (DPPH) to assess antioxidant activity, and we measured antifungal activity using mycelium growth inhibition assays with three pathogenic fungi, Magnaporthe grisea, Rhizoctonia solani, and Phytophthora infestans. Aphicidal activity against adults of Myzus persicae was determined using in vitro and in vivo assays. Spinasterol and 22,23-dihydrospinasterol exhibited moderate antioxidant activity, even at lower concentrations: 19.98% at 0.78 µg mL-1, 31.52% at 3.0 µg mL-1, 36.61% at 12.5 µg mL-1, and 49.76% at 50 µg mL-1. Spinasterol and 22,23-dihydrospinasterol showed reasonable levels of fungicidal activity toward R. solani and M. grisea, with EC50 values of 129.5 and 206.1 µg mL-1, respectively. The positive controls boscalid and carbendazim were highly effective against all fungi except boscalid for M. grisea (EC50 = 868 µg mL-1) and carbendazim for P. infestans (EC50 = 8721 µg mL-1). Significant insecticidal activity was observed in both residual and greenhouse assays, with LC50 values of 42.46, 54.86, and 180.9 µg mL-1 and 32.71, 42.46, and 173.8 µg mL-1 at 72, 48, and 24 h, respectively. The antioxidant activity of spinasterol and 22,23-dihydrospinasterol was strongly positively correlated with their antifungal and insecticidal activity. Spinasterol and 22,23-dihydrospinasterol therefore show good antioxidant and aphicidal activity with moderate fungicidal activity, making them suitable candidates for an alternative to synthetic agents.

Causes of different goiter rates with the same iodine deficiency among the pastoral and agricultural populations of Tibet: a geographical comparison.

BACKGROUND: The residents of both the agricultural and pastoral areas of Tibet share the same iodine deficiency and iodine nutrition, but the rate of thyroid goiter was significantly higher in the agricultural areas than in the pastoral areas. This project sought to determine why the populations in the iodine-deficient pastoral areas show a lower rate of thyroid goiter. METHODS: Food frequency questionnaires (FFQs) and 24 h history recalls were adopted to investigate the dietary patterns of the residents of the agricultural and pastoral areas. Meat and milk samples were collected to measure their inorganic iodine, total iodine and thyroid hormone contents using ICP-MS, AsIII-Ce4+ catalytic spectrophotometry and the Siemens' chemiluminescence method, respectively. The intake of protein, and the microelements, selenium and iron, was calculated according to their content in the food. RESULTS: The per capita daily intake of meat, dairy, and cereal in the pastoral areas was 116.7, 216.7, and 433.3 g, respectively, which are significantly higher than those in the agricultural areas (50.0, 72.2, and 375.0 g, respectively) (p < 0.05). The content of thyroid hormone in dried beef and milk in the pastoral areas was 62.6 and 13.5 µg/kg, respectively, which was significantly higher than those in the agricultural areas (25.1 and 4.1 µg/kg, respectively) (p < 0.05). The daily intake of thyroid hormone, protein and microelements, selenium and iron from foods by the residents of the pastoral areas were 10.5 µg, 99.6 g, 30.0 µg and 15.8 mg respectively, which was significantly higher than those in the agricultural areas (1.79 µg, 56.5 g, 23.8 µg and 13.2 mg, respectively) (p < 0.05). CONCLUSIONS: The significantly high intake of the food-borne thyroid hormone by the residents of the pastoral area could be the main reason the residents in the pastoral areas show a lower rate of thyroid goiter than those in the agricultural area. Moreover, the relatively high intake of protein and trace elements, selenium and iron by residents in the pastoral area could be another important factor for reducing the goiter rates.

Pharmacological Mechanisms of Tinglizi against Chronic Heart Failure Determined by Network Pharmacology and Molecular Docking.

OBJECTIVE: Tinglizi has been extensively used to treat chronic heart failure (CHF) in modern times, but the material basis and pharmacological mechanisms are still unclear. To explore the material basis and corresponding potential targets and to elucidate the mechanism of Tinglizi, network pharmacology and molecular docking methods were utilized. METHODS: The main chemical compounds and potential targets of Tinglizi were collected from the pharmacological database analysis platform (TCMSP). The corresponding genes of related action targets were queried through gene cards and UniProt database. The corresponding genes of CHF-related targets were searched through Disgenet database, and the intersection targets were obtained by drawing Venn map with the target genes related to pharmacodynamic components. Then, drug targets and disease targets were intersected and put into STRING database to establish a protein interaction network. The "active ingredient-CHF target" network was constructed with Cytoscape 3.8.2. Finally, Gene Ontology (GO) Enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment of intersection targets were analyzed using metascape. With the aid of SYBYL software, the key active ingredients and core targets were docked at molecular level, and the results were visualized by PyMOL software. Molecular docking was carried out to investigate interactions between active compounds and potential targets. RESULTS: A total of 12 active components in Tinglizi were chosen from the TCMSP database, and 193 corresponding targets were predicted. Twenty-nine potential targets of Tinglizi on CHF were obtained, of which nine were the core targets of this study. Twenty GO items were obtained by GO function enrichment analysis (P < 0.05), and 10 signal pathways were screened by KEGG pathway enrichment analysis (P < 0.05), which is closely related to the treatment of CHF by Tinglizi. The constructed drug compound composition action target disease network shows that quercetin, kaempferol, and other active compounds play a key role in the whole network. The results of molecular docking showed that all the key active ingredients, such as quercetin and isorhamnetin, were able to successfully dock with ADRB2 and HMOX1 with a total score above 5.0, suggesting that these key components have a strong binding force with the targets. CONCLUSION: Through network pharmacology and molecular docking technology, we found that the main components of Tinglizi in the treatment of CHF are quercetin, kaempferol, ß-sitosterol, isorhamnetin, and so on. The action targets are beta 2-adrenergic receptor (ADRB2), heme oxygenase 1 (HMOX1), and so on. The main pathways are advanced glycation end products/receptor for advanced glycation end products (AGE-RAGE) signaling pathway in diabetic complications, hypoxia-inducible factor (HIF-1) signaling pathway, estrogen signaling pathway, and so on. They play an integrated role in the treatment of CHF.

Ilexsaponin A1 Ameliorates Diet-Induced Nonalcoholic Fatty Liver Disease by Regulating Bile Acid Metabolism in Mice.

Bile acid (BA) metabolism is an attractive therapeutic target in nonalcoholic fatty liver disease (NAFLD). We aimed to investigate the effect of ilexsaponin A1 (IsA), a major bioactive ingredient of Ilex, on high-fat diet (HFD)-induced NAFLD in mice with a focus on BA homeostasis. Male C57BL/6J mice were fed an HFD to induce NAFLD and were treated with IsA (120 mg/kg) for 8 weeks. The results showed that administration of IsA significantly decreased serum total cholesterol (TC), attenuated liver steatosis, and decreased total hepatic BA levels in HFD-induced NAFLD mice. IsA-treated mice showed increased BA synthesis in the alternative pathway by upregulating the gene expression levels of sterol 27-hydroxylase (CYP27A1) and cholesterol 7b-hydroxylase (CYP7B1). IsA treatment accelerated efflux and decreased uptake of BA in liver by increasing hepatic farnesoid X receptor (FXR) and bile salt export pump (BSEP) expression, and reducing Na+-taurocholic acid cotransporting polypeptide (NTCP) expression. Alterations in the gut microbiota and increased bile salt hydrolase (BSH) activity might be related to enhanced fecal BA excretion in IsA-treated mice. This study demonstrates that consumption of IsA may prevent HFD-induced NAFLD and exert cholesterol-lowering effects, possibly by regulating the gut microbiota and BA metabolism.

[Content determination of eight phenols in Ilex hainanensis harvested at different time by HPLC-DAD].

This study intends to develop a high performance liquid chromatography-diode array detection(HPLC-DAD) method for simultaneous determination of chlorogenic acid, 2-hydroxymethyl-3-hydroxyl-1-butene-4-O-ß-D-(6″-O-caffeoyl)-glucopyranoside, pubescenoside B, huazhongilexone-7-O-ß-D-glucopyranoside, rutin, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C in Ilex hainanensis. The HPLC conditions are as follows: Waters XBridge C_(18 )column(4.6 mm×250 mm, 5 µm), mobile phase of 0.5% formic acid in water(A)-acetonitrile(B), gradient elution(0-8 min, 5%-12% B; 8-18 min, 12%-18% B; 18-30 min, 18%-25% B; 30-40 min, 25%-30% B; 40-42 min, 30%-80% B; 42-45 min, 80% B) at the flow rate of 0.8 mL·min~(-1), detection wavelengths of 282, 324, and 360 nm, column temperature of 25 ℃, and injection volume of 5 µL. The content of the 8 phenols in 8 samples was 0.30-6.29, 0.29-3.27, 0.15-10.4, 0.51-5.85, 0.49-9.02, 0.51-4.68, 1.93-13.4, and 0.87-5.95 mg·g~(-1), respectively. Moreover, the content of phenols in the samples collected in October was higher than that of samples harvested in other months. The established method is accurate and sensitive for the determination of phenols in I. hainanensis, which is useful for the quality improvement of this herbal medicine.

Enantioseparation and determination of triazole fungicides in fruit juice by aqueous biphasic system coupled with HPLC-MS/MS.

The aqueous biphasic system based on choline ionic liquids and ethylene/propylene oxide copolymer coupled with high-performance liquid chromatography-tandem mass spectrometry was developed for the simultaneous determination of the enantiomers of two fungicides: myclobutanil and tebuconazole. The influence of mass fractions of ionic liquids and the copolymer on the extraction efficiency of the target fungicides was investigated. The analytes are mainly concentrated in the bottom, copolymer-rich phase. The extraction efficiencies of the selected fungicides were significantly affected by the concentrations of the copolymer, and their extraction efficiencies decreased with the higher mass fractions of the copolymer, while the mass fraction of ionic liquids had little effect on their extraction behavior. Excellent extraction efficiency was achieved using the aqueous biphasic system based on choline L-lysinate and the copolymer. At three spiked concentrations of 0.01, 0.05, and 0.1 mg/kg, the average recoveries of the selected fungicides ranged from 80 to 89%, with the relative standard deviations in the range of 2.1-5.3%. Limits of quantitation for the enantiomers of tebuconazole and myclobutanil were 0.5 and 5.0 µg/kg, respectively. The developed system could be successfully applied to the analysis of triazole fungicides residue in real samples.