Pancreatic lipase immobilization on cellulose filter paper for inhibitors screening and network pharmacology study of anti-obesity mechanism.

The discovery of pancreatic lipase (PL) inhibitors is an essential route to develop new anti-obesity drugs. In this experiment, chitosan was used to add amino groups to cellulose filter paper (CFP) and then glutaraldehyde was used to covalently combine PL with amino-modified CFP through the Schiff base reaction. Under optimal immobilization conditions, CFP immobilized PL has a wide range of pH and temperature tolerance, as well as excellent reproducibility, reusability and storage stability. Subsequently, 26 natural products (NPs) were screened by immobilized PL with black tea extract having the highest inhibition rate. Three compounds with binding effects on PL (epigallocatechin gallate, theaflavin-3-gallate and theaflavin-3,3'-digallate) were captured. Molecular docking proved that these three compounds have a strong binding affinity for PL. Fluorescence spectra further revealed that theaflavin-3,3'-digallate could statically quench the intrinsic fluorescence of pancreatic lipase. The molecular docking and thermodynamic parameters indicated that electrostatic interaction was considered as the main interaction force between PL and theaflavin-3,3'-digallate. Finally, the potential anti-obesity targets and pathways of the three compounds were discussed through network pharmacology. This study not only proposes a simple and efficient method for screening PL inhibitors, but also sheds light on the anti-obesity mechanism of active compounds in black tea.

Raman spectroscopy: A promising analytical tool used in human reproductive medicine.

Over the past few years, there has been growing interest in developing new methods of embryo quality assessment to improve the outcomes of assisted reproductive technologies in the medical field. Raman microscopy as an increasingly promising analytical tool has been widely used in life sciences, biomedicine and "omics" to study molecular, biochemical components, living cells and tissues due to the label-free and non-destructive nature of the imaging technique. This paper reviews the analytical capability of Raman microscopy and applications of Raman spectroscopy technology mainly in reproductive medicine. The purpose of this review is to introduce the Raman spectroscopy technology, application and underlying principles of the method, to provide an intact picture of its uses in biomedical science and reproductive medicine, to offer ideas for its future application, verification and validation. The focus is on the application of Raman spectroscopy in the reproductive medicine field, including the application in gametes, embryos and spent embryo culture media.

How do autistic severity and family functioning influence parental stress in caregivers of children with autism spectrum disorder in China? The important role of parental self-efficacy.

Introduction: Parental stress among primary caregivers of children with autism spectrum disorder (ASD) is a significant concern. While previous research indicates that both family and child factors substantially influence parental stress, a few studies have comprehensively examined these factors from family, parent, and child perspectives. Moreover, the psychological mechanisms underlying parental stress remain underexplored. Method: This study obtained a valid sample of 478 primary caregivers of children diagnosed with ASD in China and employed mediation and moderated mediation analyses to investigate the relationships between family adaptability and cohesion (FAC), ASD severity, parental self-efficacy, and parental stress. Result: Results revealed that higher FAC was linked to reduced parental stress through increased parental self-efficacy. The indirect effect of parental self-efficacy was more substantial for caregivers of children with severe symptoms than those with mild symptoms. Discussion: These findings offer insights into how FAC influences parental stress and underscore the importance of parental self-efficacy as a coping resource for mitigating parental stress. This study provides valuable theoretical and practical implications for understanding and addressing parental stress, particularly in families raising children with ASD.

Efficacy and safety of montelukast adjuvant therapy in adults with cough variant asthma: A systematic review and meta-analysis.

BACKGROUND: Montelukast is a highly selective and specific cysteinyl leukotriene receptor antagonist used in the treatment of asthma. Whether montelukast as adjuvant therapy can significantly and safely treat adults with cough variant asthma (CVA) remains inconclusive. AIMS: This meta-analysis systematically evaluated the efficacy and safety of montelukast as an adjuvant treatment for adults with CVA. MATERIALS AND METHODS: Randomized controlled trials (RCTs) on montelukast combined with inhaled corticosteroids (ICS) and long-acting ß2 agonists (LABAs) to treat CVA in adults, from inception to March 6, 2023, were retrieved from the CNKI, Wanfang, VIP, CBM, PubMed, Embase, Cochrane Library, and Web of Science databases and Clinical Trials website. Review Manager (version 5.4) and Stata (version 15.0) were used to conduct the meta-analysis. RESULTS: A total of 15 RCTs were ultimately included in the meta-analysis. It was established that montelukast as adjuvant therapy raised the total effective rate (RR = 1.20, 95% confidence interval [CI] [1.13, 1.27], P < 0.01) and improved the FEV1% (SMD = 0.91, 95% CI [0.40, 1.41], P < 0.01), PEF% (SMD = 0.63, 95% CI [0.38, 0.88], P < 0.01), FEV1 (SMD = 1.15, 95% CI [0.53, 1.77], P < 0.01), PEF (SMD = 0.64, 95% CI [0.42, 0.86], P < 0.01), and FEV1/FVC% (SMD = 0.76, 95% CI [0.51, 1.01], P < 0.01) and reduced the recurrence rate (RR = 0.28, 95% CI [0.15, 0.53], P < 0.01). The incidence of adverse reactions was higher in the montelukast auxiliary group compared to the control group but with no statistical difference (RR = 1.32, 95% CI [0.89, 1.96], P = 0.17). CONCLUSION: Existing evidence indicated that the use of montelukast as an adjuvant therapy had therapeutic efficacy superior to ICS + LABA alone for the treatment of adult patients with CVA. However, further research is needed, especially a combination of high-quality long-term prospective studies and carefully designed RCTs.

Indoleamine 2,3 dioxygenase 1 immobilization on magnetic nanoparticles for screening inhibitors from coffee.

In this study, a ligand fishing method was developed to screen potential indoleamine 2,3-dioxygenase 1 (IDO1) inhibitors from coffee extracts by immobilization of IDO1 enzyme on amino-modified magnetic nanoparticles combined with UHPLC-Q-TOF-MS/MS analysis. Parameters including enzyme concentration, immobilization time, the pH of glutaraldehyde and the amount of magnetic nanoparticles were optimized. The results indicated that immobilized IDO1 could be reused 5 times and was stable during storage for 7 days. Several IDO1 ligands were captured by incubating immobilized IDO1 with coffee extract, of which 10 showed an obvious difference comparing to non-conjugated bare nanoparticles. In vitro inhibitory activity was further performed by CE analysis, in which ferulic acid and chlorogenic acid had better IDO1 inhibitory activity, with IC50 value of 113.7 µM and 307.5 µM. These results demonstrate that this method provides an effective platform for identifying and screening IDO1 inhibitors from natural products.

Off-line and on-line liquid chromatography-mass spectrometry methods with immobilized bio-macromolecules for drug screening from natural sources.

Natural products are an important source of major compounds in drug discovery. Currently, rapid screening and identification of bioactive compounds is a challenge due to the complicated chemical composition of natural products. Affinity screening methods based on liquid chromatography coupled with mass spectrometry have seen increasing interest in the past few years. In this review, the various strategies are classified into off-line and on-line modes. The principles and applications of these screening methods such as magnetic nanoparticles, affinity solid-phase extraction, immobilized liposome chromatography and cell membrane chromatography are introduced. This review covers the research advances of LC-MS based screening methods from 2019 to mid 2022, discussing their advantages and disadvantages, and providing an outlook for the future of this field.

SWL-1 Reverses Fluconazole Resistance in Candida albicans by Regulating the Glycolytic Pathway.

Candida albicans is a ubiquitous clinical fungal pathogen. Prolonged use of the first-line antifungal agent fluconazole (FLC) has intensified fungal resistance and limited its effectiveness for the treatment of fungal infections. The combined administration of drugs has been extensively studied and applied. SWL-1 is a lignin compound derived from the Traditional Chinese Medicine Schisandra chinensis. In this study, we show that SWL-1 reverses resistance to fluconazole in C. albicans when delivered in combination, with a sharp decrease in the IC50 of fluconazole from >200 to 3.74 ± 0.25 µg/ml, and also reverses the fluconazole resistance of C. albicans in vitro, with IC50 from >200 to 5.3 ± 0.3 µg/ml. Moreover, killing kinetics curves confirmed the synergistic effects of fluconazole and SWL-1. Intriguingly, when SWL-1 was administered in combination with fluconazole in a mouse model of systemic infection, the mortality of mice was markedly decreased and fungal colonization of the kidney and lung was reduced. Further mechanistic studies showed that SWL-1 significantly decreased intracellular adenosine 5'-triphosphate (ATP) levels and inhibited the function of the efflux pump responsible for fluconazole resistance of C. albicans. Proteomic analysis of the effects of SWL-1 on C. albicans showed that several enzymes were downregulated in the glycolytic pathway. We speculate that SWL-1 significantly decreased intracellular ATP levels by hindering the glycolysis, and the function of the efflux pump responsible for fluconazole resistance of C. albicans was inhibited, resulting in restoration of fluconazole sensitivity in FLC-resistant C. albicans. This study clarified the effects and mechanism of SWL-1 on C. albicans in vitro and in vivo, providing a novel approach to overcoming fungal resistance.

[Wenweishu Capsule alleviates gastric mucosal lesion in rats with chronic gastritis by inhibiting NF-κB pathway].

Objective To investigate the effect of Wenweishu Capsule on the expression of nuclear factor kappa B (NF-κB)-related proteins in chronic gastritis model rats. Methods Wistar rat models of chronic gastritis were constructed by alternant administrations of sodium deoxycholate, ammonia, alcohol solution and the hunger disorder method. The rats were randomly divided into control group, model group, vatacoenayme group, high-, middle- and low-dose Wenweishu Capsule groups. The control group and model group were treated with normal saline (2 mL/d). The other groups were separately treated with 0.3 g/kg vatacoenayme and 0.76, 0.38, 0.19 g/kg Wenweishu Capsule for 4 weeks. Naked eye observation and HE staining were used to evaluate the pathological changes of gastric tissue. ELISA was performed to measure the levels of tumor necrosis factor α (TNF-α) and interleukin 6 (IL-6) in the serum. Immunofluorescence staining was employed to observe the expression of NF-κBp65, inhibitor of NF-κBα (IκBα) in the gastric tissue. Simple Western was utilized to detect the protein levels of NF-κBp65, IκBα and COX2 in the gastric tissue. Results Compared with the control group, the model group was found with thinner gastric mucosa, disappeared or shallower folds, obviously infiltrated mucosa inflammatory cells, disordered glands, and significantly increased levels of serum inflammatory cytokines and NF-κBp65, IκBα and COX2 proteins in the gastric tissue. Compared with the model group, the vatacoenayme group, high- and middle-dose Wenweishu Capsule groups showed the alleviated gastric cavity signs, improved histopathological changes, reduced levels of TNF-α and IL-6 in the serum, and decreased expression of NF-κBp65, IκBα and COX2 proteins in the gastric tissue. Conclusion Wenweishu Capsule can reduce the levels of serum inflammatory factors by inhibiting NF-κB pathway in the gastric tissue, so as to alleviate the injury of gastric mucosa in rats with chronic gastritis.

What Is the Biological Function of Uric Acid? An Antioxidant for Neural Protection or a Biomarker for Cell Death.

The main aim of the present study was to investigate the biological function of uric acid. The level of uric acid in different organs in normal male rats was determined with uric acid assay kits, and the expression level of genes in the organs was determined by RNA quantitative sequencing. The correlation analysis between uric acid in the organs and gene expression (measured by FPKM value) was made. Serum uric acid (SUA) in patients with breast cancer or with breast benign tumor was assayed when the diagnosis was made, and SUA in patients with breast cancer was also assayed just after chemotherapy. There were 1937 mRNAs whose expression level significantly correlated with the level of uric acid, and most of which were associated with purine or nucleoside metabolism, cellular metabolism, cell cycles, and cell death pathways. Further analysis showed that the level of uric acid was highly correlated with cell death rather than cell viability. The level of SUA in patients with breast cancer was higher than that in patients with breast benign tumor, and the SUA increased after chemotherapy. All the results suggested that uric acid was mainly synthesized from local nucleosides degraded from dead cells, and uric acid could be an important biomarker for cell death rather than an antioxidant for neural protection.

Intestinal tract is an important organ for lowering serum uric acid in rats.

The kidney was recognized as a dominant organ for uric acid excretion. The main aim of the study demonstrated intestinal tract was an even more important organ for serum uric acid (SUA) lowering. Sprague-Dawley rats were treated normally or with antibiotics, uric acid, adenine, or inosine of the same molar dose orally or intraperitoneally for 5 days. Rat's intestinal tract was equally divided into 20 segments except the cecum. Uric acid in serum and intestinal segment juice was assayed. Total RNA in the initial intestinal tract and at the end ileum was extracted and sequenced. Protein expression of xanthine dehydrogenase (XDH) and urate oxidase (UOX) was tested by Western blot analysis. The effect of oral UOX in lowering SUA was investigated in model rats treated with adenine and an inhibitor of uric oxidase for 5 days. SUA in the normal rats was 20.93±6.98 µg/ml, and total uric acid in the intestinal juice was 308.27±16.37 µg, which is two times more than the total SUA. The uric acid was very low in stomach juice, and attained maximum in the juice of the first segment (duodenum) and then declined all the way till the intestinal end. The level of uric acid in the initial intestinal tissue was very high, where XDH and most of the proteins associated with bicarbonate secretion were up-regulated. In addition, SUA was decreased by oral UOX in model rats. The results suggested that intestinal juice was an important pool for uric acid, and intestinal tract was an important organ for SUA lowering. The uric acid distribution was associated with uric acid synthesis and secretion in the upper intestinal tract, and reclamation in the lower.