Paclobutrazol Residue Determination in Potato and Soil Using Low Temperature Partition Extraction and Ultrahigh Performance Liquid Chromatography Tandem Mass Spectrometry

A simple, accurate, and highly sensitive analytical method was developed for determining the paclobutrazol residue in potato and soil, the dynamics dissipation in soil. Extraction was carried out by low temperature partitioning and analyzed by ultrahigh performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS). For a favor extraction yield, the parameters such as temperature and solvent were optimized. The result showed that sample would be easily frozen and separated using acetonitrile under −20°C for 10 min. The limit of detection (LOD) was 0.5 μg/kg, and the limit of quantification (LOQ) was 2 and 5 μg/kg for potato and soil, respectively. The influence of paclobutrazol residue in potato was evaluated. The possible contamination of paclobutrazol from surface can be rinsed by distilled water or peeled off, but the paclobutrazol in potato harvest comes mainly from absorption and transport, which could not be removed by peeling. The half-life of paclobutrazol in soil was 20.64 days, and the residue was below 0.22 mg/kg on 50th day after spraying. According to the risk assessment with Need Maximum Daily Intake (NEDI) and Acceptable Daily Intake (ADI), a Maximum Residue Limit (MRL) of paclobutrazol in potato was recommended as 1.0 mg/kg

Study on the Mechanism of miR-520d-5p and LMO4 in Regulating Thyroid Cancer Progression

The pathogenesis of thyroid cancer is closely related to environmental factors. Gene mutation and molecular biological changes of thyroid tissue caused by environmental changes are one of the important factors inducing thyroid cancer. Although the molecular mechanism of thyroid cancer is still not fully elucidated, with the development of molecular biology technology, more and more thyroid cancer-specific genetic changes and molecular markers have been excavated. This article systematically summarizes the current research progress of miR-520d-5p and LMO4 in thyroid cancer, and discusses how they participate in the regulation of the biological behavior of tumor cells and potential molecular signaling pathways, so as to provide new theoretical basis and ideas for the precise diagnosis and treatment of thyroid cancer

Astragaloside IV improves slow transit constipation by regulating gut microbiota and enterochromaffin cells

Purpose: Slow transit constipation (STC) is a common gastrointestinal disorder characterized by altered gut microbiota and reduced number of enterochromaffin cells (ECs). Astragaloside IV (AS-IV), a low drug permeability saponin, has showed beneficial effects on patients with STC. However, the specific mechanism by which AS-IV regulates STC remains unclear. In this study, we aimed to investigate the effect of AS-IV on STC and its associated mechanisms involving gut microbiota.Methods: The effect of AS-IV on STC was evaluated on STC mice induced with loperamide. We measured defecation frequency, intestinal mobility, ECs loss, and colonic lesions in STC mice treated with AS-IV. We also analyzed the changes in gut microbiota and metabolites after AS-IV treatment. Moreover, we investigated the relationship between specific gut microbes and altered fecal metabolites, such as 3-bromotyrosine (3-BrY). We also conducted in vitro experiments to investigate the effect of 3-BrY on caspase-dependent apoptosis of ECs and the activation of the p38 MAPK and ERK signaling pathways induced by loperamide.Results: AS-IV treatment promoted defecation, improved intestinal mobility, suppressed ECs loss, and alleviated colonic lesions in STC mice. AS-IV treatment also affected gut microbiota and metabolites, with a significant correlation between specific gut microbes and altered fecal metabolites such as 3-BrY. Furthermore, 3-BrY may potentially reduce caspase-dependent apoptosis of ECs and protect cell survival by inhibiting the activation of the p38 MAPK and ERK signaling pathways induced by loperamide.Conclusion: Our findings suggest that changes in gut microbiota and ECs mediated the therapeutic effect of STC by AS-IV. These results provide a basis for the use of AS-IV as a prebiotic agent for treating STC. The specific mechanism by which AS-IV regulates gut microbiota and ECs warrants further investigation

Single-cell RNA sequencing reveals cellular dynamics and therapeutic effects of astragaloside IV in slow transit constipation

The cellular characteristics of intestinal cells involved in the therapeutic effects of astragaloside IV (AS-IV) for treating slow transit constipation (STC) remain unclear. This study aimed to determine the dynamics of colon tissue cells in the STC model and investigate the effects of AS-IV treatment by single-cell RNA sequencing (scRNA-seq). STC mouse models were developed using loperamide, with subsequent treatment using AS-IV. Colon tissues and feces were collected for scRNA-seq and targeted short-chain fatty acid quantification. We integrated scRNA-seq data with network pharmacology to analyze the effect of AS-IV on constipation. AS-IV showed improvement in defecation for STC mice induced by loperamide. Notably, in STC mice, epithelial cells, T cells, B cells, and fibroblasts demonstrated alterations in cell proportions and aberrant functions, which AS-IV partially rectified. AS-IV has the potential to modulate the metabolic pathway of epithelial cells through its interaction with peroxisome proliferator-activated receptor gamma (PPARγ). AS-IV reinstated fecal butyrate levels and improved energy metabolism in epithelial cells. The proportion of naïve CD4+T cells is elevated in STC, and the differentiation of these cells into regulatory T cells (Treg) is regulated by B cells and fibroblasts through the interaction of ligand-receptor pairs. AS-IV treatment can partially alleviate this trend. The status of fibroblasts in STC undergoes alterations, and the FB_C4_Adamdec1 subset, associated with angiogenesis and the Wingless-related integration (Wnt) pathway, emerges. Our comprehensive analysis identifies perturbations of epithelial cells and tissue microenvironment cells in STC and elucidates mechanisms underlying the therapeutic efficacy of AS-IV

Expression of the phosphorylated MEK5 protein is associated with TNM staging of colorectal cancer

<p>Abstract</p> <p>Background</p> <p>Activation of MEK5 in many cancers is associated with carcinogenesis through aberrant cell proliferation. In this study, we determined the level of phosphorylated MEK5 (pMEK5) expression in human colorectal cancer (CRC) tissues and correlated it with clinicopathologic data.</p> <p>Methods</p> <p>pMEK5 expression was examined by immunohistochemistry in a tissue microarray (TMA) containing 335 clinicopathologic characterized CRC cases and 80 cases of nontumor colorectal tissues. pMEK5 expression of 19 cases of primary CRC lesions and paired with normal mucosa was examined by Western blotting. The relationship between pMEK5 expression in CRC and clinicopathologic parameters, and the association of pMEK5 expression with CRC survival were analyzed respectively.</p> <p>Results</p> <p>pMEK5 expression was significantly higher in CRC tissues (185 out of 335, 55.2%) than in normal tissues (6 out of 80, 7.5%; <it>P </it>< 0.001). Western blotting demonstrated that pMEK5 expression was upregulated in 12 of 19 CRC tissues (62.1%) compared to the corresponding adjacent nontumor colorectal tissues. Overexpression of pMEK5 in CRC tissues was significantly correlated to the depth of invasion (<it>P </it>= 0.001), lymph node metastasis (<it>P </it>< 0.001), distant metastasis (<it>P </it>< 0.001) and high preoperative CEA level (<it>P </it>< 0.001). Consistently, the pMEK5 level in CRC tissues was increased following stage progression of the disease (<it>P </it>< 0.001). Analysis of the survival curves showed a significantly worse 5-year disease-free (<it>P </it>= 0.002) and 5-year overall survival rate (<it>P </it>< 0.001) for patients whose tumors overexpressed pMEK5. However, in multivariate analysis, pMEK5 was not an independent prognostic factor for CRC (DFS: <it>P </it>= 0.139; OS: <it>P </it>= 0.071).</p> <p>Conclusions</p> <p>pMEK5 expression is correlated with the staging of CRC and its expression might be helpful to the TNM staging system of CRC.</p

Platelet Abnormalities after Splenectomy for Hypersplenism in Decompensated Cirrhosis: A Case Report

Patients in the decompensated stage of cirrhosis are usually associated with hepatic decompensation and portal hypertension, splenomegaly, hypersplenism, and abnormal or significantly reduced blood counts in at least one of the three blood cell lines, mainly platelets. At present, surgical removal of the spleen is still an essential treatment for hypersplenism. The majority of patients who undergo splenectomy can effectively improve the decrease in peripheral blood platelet count. In order to provide some reference value for the diagnosis and treatment of similar clinical cases in the future, we report a case of platelet reduction in hypersplenism in cirrhosis after splenectomy, in which platelets first rose to normal and then decreased abnormally.</jats:p

Development and Validation of Multiclass Chiral Pesticide Residues Analysis Method in Tea by QuEChERS Combined Liquid Chromatography Quadruple-Linear Ion Trap Mass Spectrometry

Abstract Background No single pure enantiomeric pesticide residues was investigated and set regulations for tea quality safety and risk assessment. Objective Due to lack of chiral pesticide analysis method and data, the Maximum residue limits (MRLs) about the chiral pesticides in tea was unknown. Method An analytical method for the determination of chiral pesticide residues by QuEChERS combined chiral liquid chromatography quadruple/linear ion trap mass spectrometry (LC-MS/MS-Qtrap) was developed and applied to the analysis of various teas. Results The mean recoveries for pesticides enantiomers ranged from 75.9% to 112.4%. Reproducibility represented by relative standard deviation percentage was 10% or less. Good linearity was achieved for all enantiomers with determination coefficients (r2) greater than 0.99. The detection of limit (CCα) and quantification of limit (CCβ) were 0.2 ∼1 µg/kg and 0.5∼5 µg/kg, respectively. Conclusions The method was suitable for monitoring the enantiomeric pesticide residues in various teas. Highlights Enantioselective multiclass pesticide residues were determined in various teas by LC-MS/MS-Qtrap, additional Qtrap scan functions greatly enhance the performance of screening, confirmation, and identification of chiral pesticides. </jats:sec