Ultra-performance Liquid Chromatography Quadrupole Time-of-flight Mass Spectrometry Research Articles

Diterpenoids from Scutellaria barbata D. Don Exert the Antiproliferative Effect on Colorectal Cancer Cells Via Hedgehog Signaling Pathway

Background: Scutellaria barbata D. Don, also known as “Banzhilian” in China, the diterpenes of Banzhilian (Bzl-D) is a commonly used herbal medicine for treating various tumors including colorectal cancer (CRC). Objectives: This study aimed to examine the function of Bzl-D in inhibiting CRC development and explore its potential targets and mechanisms involving the Hedgehog (HH) pathway. Materials and methods: To determine the major components in Bzl-D, we performed ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-qTOF-MS) analysis. The proliferation, migration, invasion, and apoptosis abilities of the CRC HT-29 cell line were examined, and the related protein expression was tested. Tumorigenesis after treatment was detected by in vivo experiments. Results: Nineteen diterpenoids were detected, Bzl-D significantly inhibited the cell proliferation and metastasis and induced the apoptosis of HT-29 cells through binding to Sonic HH (SHH) and down-regulating the key proteins (SHH, patched, smoothened, and glioma-associated oncogene) of the HH pathway, thus. Of note, Bzl-D remarkably suppressed the growth of HT-29 xenografts in nude mice. Conclusion: This research demonstrates that Bzl-D is a promising anti-rectal cancer drug, providing a new entry point for targeted therapy of CRC.

Non-targeted metabolomics of spleen and liver metabolism in mice treated with Pruni Semen processed with different methods

Ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS) was employed to investigate the impacts of Pruni Semen processed with different methods(raw and fried) on the liver and spleen metabolism in mice. A total of 24 male mice were randomly assigned to three groups: raw Pruni Semen group, fried Pruni Semen group, and control(deionized water) group. Mice in the three groups were orally administrated with 0.01 g·mL~(-1) Pruni Semen decoction or deionized water for one week. After that, the liver and spleen tissues were collected, and liquid chromatography-mass spectrometry(LC-MS)-based metabolomic analysis was carried out to investigate the impact of Pruni Semen on the liver and spleen metabolism in mice. Compared with thte control group, the raw Pruni Semen group showed up-regulation of 11 metabolites and down-regulation of 57 metabolites in the spleen(P<0.05), as well as up-regulation of 15 metabolites and down-regulation of 58 metabolites in the liver(P<0.05). The fried Pruni Semen group showed up-regulation of 31 metabolites and down-regulation of 10 metabolites in the spleen(P<0.05), along with up-regulation of 26 metabolites and down-regulation of 61 metabolites in the liver(P<0.05). The differential metabolites identified in the raw Pruni Semen group were primarily associated with alanine, aspartate, and glutamate metabolism, purine metabolism, amino sugar and nucleotide sugar metabolism, and D-glutamine and D-glutamate metabolism. The differential metabolites identified in the fried Pruni Semen group predominantly involved riboflavin metabolism, amino sugar and nucleotide sugar metabolism, purine metabolism, alanine, aspartate, and glutamate metabolism, D-glutamine and D-glutamate metabolism, and glutathione metabolism. The findings suggest that both raw and fried Pruni Semen have the potential to modulate the metabolism of the liver and spleen in mice by influencing the glutamine and glutamate metabolism.

Effects of the combination of Epimedii Folium and Ligustri Lucidi Fructus on apoptosis and autophagy in SOP rats and osteoblasts via PI3K/AKT/mTOR pathway

BackgroundThis study aimed to investigate the effects of the combination of Epimedii Folium (EF) and Ligustri Lucidi Fructus (LLF) on regulating apoptosis and autophagy in senile osteoporosis (SOP) rats. MethodsFirstly, we identified the components in the decoction and drug-containing serum of EL (EF&LLF) by Ultra performance liquid chromatography-quadrupole-time of flight-mass spectrometry (UPLC-Q-TOF-MS). Secondly, SOP rats were treated with EF, LLF, EL and caltrate to evaluate the advantages of EL. Finally, H2O2-, chloroquine-, and MHY1485-induced osteoblasts were treated with different doses of EL to reveal the molecular mechanism of EL. We detected bone microstructure, oxidative stress levels, ALP activity and the expressions of Bax, Bcl-2, caspase3, P53, Beclin-1, p-PI3K, PI3K, p-Akt, Akt, p-mTOR, mTOR, and LC3 in vivo and in vitro. Results36 compounds in EL decoction and 23 in EL-containing serum were identified, including flavonoids, iridoid terpenoids, phenylethanoid glycosides, polyols and triterpenoids. EL could inhibit apoptosis activity and increase ALP activity. In SOP rats and chloroquine-inhibited osteoblasts, EL could improve bone tissue microstructure and osteoblasts functions by upregulating Bcl-2, Beclin1, and LC3-II/LC3-I, while downregulating p53 in all treatment groups. In H2O2-induced osteoblasts, EL could upregulate the protein and mRNA expressions of Bcl-2 while downregulate LC3-II/LC3-I, p53 and Beclin1. Besides, EL was able to down-regulate PI3K/AKT/mTOR pathway which activated in SOP rats and MHY1485-induced osteoblasts. ConclusionsThese findings demonstrate that EL with bone protective effects on SOP rats by regulating autophagy and apoptosis via PI3K/Akt/mTOR signaling pathway, which might be an alternative medicine for the treatment of SOP.

Plasma metabolomics for diagnostic biomarkers on ectopic pregnancy

Metabolomics is a relatively novel omics tool to provide potential biomarkers for early diagnosis of the diseases and to insight the pathophysiology not having discussed ever before. In the present study, an ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) was employed to the plasma samples of Group T1: Patients with ectopic pregnancy diagnosed using ultrasound, and followed-up with beta-hCG level (n = 40), Group T2: Patients with ectopic pregnancy diagnosed using ultrasound, underwent surgical treatment and confirmed using histopathology (n = 40), Group P: Healthy pregnant women (n = 40) in the first prenatal visit of pregnancy, Group C: Healthy volunteers (n = 40) scheduling a routine gynecological examination. Metabolite extraction was performed using 3 kDa pores - Amicon® Ultra 0.5 mL Centrifugal Filters. A gradient elution program (mobile phase composition was water and acetonitrile consisting of 0.1% formic acid) was applied using a C18 column (Agilent Zorbax 1.8 μM, 100 x 2.1 mm). Total analysis time was 25 min when the flow rate was 0.2 mL/min. The raw data was processed through XCMS – R program language edition where the optimum parameters detected using Isotopologue Parameter Optimization (IPO). The potential metabolites were identified using MetaboAnalyst 5.0 and finally 27 metabolites were evaluated to be proposed as potential biomarkers to be used for the diagnosis of ectopic pregnancy.

Mechanism of "Trichosanthis Fructus-Allii Macrostemonis Bulbus" in treating cardiovascular diseases with syndrome of combined phlegm and stasis based on serum metabolomics

This study aimed at investigating the mechanism of Trichosanthis Fructus-Allii Macrostemonis Bulbus(GX) in treating cardiovascular diseases in rats with the syndrome of combined phlegm and stasis. The rat model was established by a high-fat diet, ice-water bath combined with subcutaneous injection of adrenalin hydrochloride, and the syndrome score was determined. The serum samples of rats in the control, model, and GX groups were collected. Ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS) was employed to analyze the metabolic profiles of the serum samples. The differential metabolites were screened and identified by partial least squares-discriminant analysis(PLS-DA) and orthogonal partial least squares-discriminant analysis(OPLS-DA). The intervention targets of GX-regulated metabolites and their metabolic pathways were searched against MetaboAnalyst. Gene Ontology enrichment was carried out to predict the biological pathways associated with the intervention targets of metabolic pathways. A total of 129 potential biomarkers were detected in the rat model with the syndrome of combined phlegm and stasis via metabolomics, and GX regulated 54 metabolites in several metabolic pathways such as linoleic acid metabolism, sphingolipid metabolism, and tricarboxylic acid cycle. The further screening against MetaboAnalyst showed that GX recovered the levels of nine metabolites associated with cardiovascular diseases with the syndrome of combined phlegm and stasis, which involved 69 targets in the pathways regarding cholesterol metabolism, fatty acid metabolism, inflammatory response, and glucose homeostasis and metabolism. The above-mentioned results suggested that GX can alleviate the symptoms of the rat model of cardiovascular diseases with the syndrome of combined phlegm and stasis by regulating the metabolism of linoleic acid, sphingosine, docosahexaenoic acid, rosemary acid, succinic acid, adenine, L-phenylalanine, L-valine and modulating the biological pathways such as cholesterol metabolism, fatty acid metabolism, inflammatory response, and glucose homeostasis and metabolism.

Phytochemical components analysis and hypolipidemic effect on hyperlipidemia mice of the aerial parts from Allium sativum.

The bulbs of Allium sativum are widely used as food or seasoning (garlic), while they have also been utilized as a famous traditional medicine since ancient eras for the treatment of scabies, tuberculosis, pertussis, diarrhea and dysentery, etc. However, very few studies focus on their abundant aerial parts, which are normally discarded during the harvest season. The hyperlipidemic mice model has been used to study the lipid-lowering effect of the aerial parts in this article. 180 mice were randomly divided into 18 groups, including blank control (BC), model (Mod), positive control (PC), and low-, medium-, and high-dose groups of the crude extract, petroleum ether, ethyl acetate, n-butanol, and residual water extracts (corresponding to CE, PEE, EAE, NBE, WE), with 10 mice in each group. The preventive effects of the extracts on hyperlipidemic mice lasted for four weeks. Ultra performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q/TOF-MS) and gas chromatography tandem mass spectrometry (GC-MS/MS) were used to analyze the chemical components of NBE and PEE respectively. The results of the mice experiment showed that n-butanol extract (NBE) and petroleum ether extract (PEE) from the aerial parts could significantly reduce the contents of total cholesterol (TC), triglycerides (TG), low density lipoprotein cholesterol (LDL-C), alanine transaminase (ALT) and aspartate transaminase (AST) in serum of hyperlipidemic mice, and increase the contents of high density lipoprotein cholesterol (HDL-C). They could enhance the activity of superoxide dismutase (SOD) in liver and reduce the level of malondialdehyde (MDA). At the same time, they could improve steatosis and inflammation of liver cells. The results of phytochemical components analysis showed that NBE was rich in organic acids, flavonoids and nitrogen-containing constituents, while PEE contained organic sulfur compounds, aliphatic acids and derivatives, alkaloids, phytosterols, etc. These results support that the aerial parts of A. sativum are an interesting source of bioactive ingredients that may be useful in the prevention and treatment of hyperlipidemia.

Plant-microbial interactions inspired the discovery of novel sesquiterpenoid dimeric skeletons of hidden natural products from Hibiscus tiliaceus

Hidden natural products are representative of defensive strategies produced in vivo in diseased plants, a process that is induced by the plant immune system. The first transcriptome library of uninfected and pathogen infected Hibiscus tiliaceus stems was constructed by transcriptome sequencing technology, genes related to cadinene-type sesquiterpenoid biosynthesis were screened and combined with ultra-performance liquid chromatography-quadrupole-time of flight mass spectrometry (UPLC-QTOF-MS) analysis data, which indicated pathological tissue had potential to produce novel carbon skeletons of cadinane sesquiterpenoid dimers. Successfully, two cadinane-derived sesquiterpenoid dimers with unprecedented carbon skeletons, hibisceusanols A (1) and B (2) were isolated for the first time from the stems of H. tiliaceus induced by plant-microbial interactions. Their structures and absolute configurations were unambiguously established by spectroscopy, advanced chemistry development (ACD) and electronic circular dichroism (ECD) methods. Compounds 1 and 2 exhibited significant antitumor activity in vitro with half maximal inhibitory concentration (IC50) values of 2.3–7.2 µmol/L. The anticancer effect was generated via the induction of HepG2 cell apoptosis by inhibiting the phosphatidylinositol 3-kinase (PI3K) pathway.

Integrated non-targeted metabolomics and network pharmacology to reveal the mechanisms of berberine in the long-term treatment of PTZ-induced epilepsy

AimsThe increasing resistance to anti-seizure medications (ASMs) and the ambiguous mechanisms of epilepsy highlight the pressing demand for the discovery of pioneering lead compounds. Berberine (BBR) has received significant attention in recent years within the field of chronic metabolic disorders. However, the reports on the treatment of epilepsy with BBR are not systematic and the mechanism remains unclear. Main methodsIn this study, the seizure behaviors of mice were recorded following subcutaneous injection of pentetrazol (PTZ). Non-targeted metabolomics was used to analyze the serum metabolites based on ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS). Meanwhile, multivariate statistical methods were used for metabolite identification and pathway analysis. Furthermore, network pharmacology, molecular docking, and quantitative real-time PCR assay were used for the target identification. Key findingsBBR had anti-seizure effects on PTZ-induced seizure mice after long-term treatment. Tryptophan metabolism and phenylalanine metabolism were involved in regulating the therapeutic effects of BBR. SignificanceThis study reveals the potential mechanism of BBR for epilepsy treatment based on non-targeted metabolomics and network pharmacology, which provides evidence for uncovering the pathogenesis of epilepsy, suggesting that BBR is a potential lead compound for anti-epileptic treatment.

Metabolomics study of the hepatoprotective effects and mechanism of aqueous extract of Dendrobium nobile Lindl. on alcohol liver injury in rats.

Dendrobium nobile Lindl. (DNL) is effective for the treatment of alcohol liver disease (ALD), but the underly mechanism is still unclear. This research aimed to investigate the effects and mechanism of aqueous extract of Dendrobium nobile Lindl (AEDNL) in ALD rats based on a metabolomics approach. In this study, 18 Sprague-Dawley male rats were randomly divided into control, model, and AEDNL groups (n=six). Rats in the AEDNL group were given AEDNL (152 mg/kg) intragastric administration from the first day for 30 consecutive days. From day 15 to day 30, model and AEDNL groups were given 30% ethanol (10 ml/kg) after 4 h of daily administration. Then, serum and liver samples were collected for biochemical analysis, histopathological examination, and Ultra Performance Liquid Chromatography-Quadrupole Time-of-Flight Mass Spectrometry (UPLC-Q-TOF/MS) determination for metabolomic analysis. Compared with the model group, the liver/body weight index and serum levels of TC, LDL-C, and TBIL in the AEDNL group were significantly decreased. Hepatocyte cord arrangement, hepatocyte balloon, and fat vacuolization were significantly improved in the AEDNL group. Metabolism profiles were changed in the model and AEDNL groups. Seven and two common differential metabolites (Guanosine3',5'-cyclic monophosphate, and Glutaric acid) were found in serum and liver, respectively. In addition, the hepatoprotective effect of AEDNL on ALD was related to steroid hormone biosynthesis, riboflavin metabolism, and glycerophospholipid metabolism. The research could provide novel evidence of the protective effects of AEDNL on ALD.

Determination of seven mycotoxins in cereals by ultra performance liquid chromatography-quadrupole-time of flight mass spectrometry based on the self-built database

A method for the determination of seven mycotoxins in rice and wheat by ultra performance liquid chromatography-quadrupole-time of flight mass spectrometry (UPLC-Q-TOF/MS) based on self-built database was established. Samples were extracted with 0.2% formic acid aqueous solution-acetonitrile (50∶50, v/v), dehydrated and salted according to the QuEChERS method (4 g of magnesium sulfate, 1 g of sodium chloride, 1 g of sodium citrate, 0.5 g of citrate disodium salt), and separated on an HSS T3 column (100 mm×2.1 mm, 1.8 μm). UPLC-Q-TOF/MS with MSE screening was performed, and the positive and negative ions of the screened mycotoxins were calibrated and quantified using matrix-matched standard curves with time of flight multiple reaction monitoring (TOF-MRM). The results showed that aflatoxin G1 (AFG1), aflatoxin G2 (AFG2), aflatoxin B1 (AFB1), aflatoxin B2 (AFB2), and ochratoxin A (OTA) exhibited moderate matrix effects in rice, while OTA and zearalenone (ZEN) exhibited moderate matrix effects in wheat. The seven mycotoxins showed good linearities in their respective concentration ranges, with correlation coefficients (r) of 0.9900-0.9998. The limits of detection (LODs) for rice and wheat were 0.50-400 and 0.50-200 μg/kg, respectively, and the limits of quantification (LOQs) for both cereals were 1.00-800 μg/kg. In rice, the average recoveries at three spiked levels of low, medium, and high were 88.1%-123.9%, with relative standard deviations (RSDs) of 0.2%-13.6%. In wheat, the average recoveries at three spiked levels of low, medium, and high were 102.0%-123.4%, with RSDs of 0.8%-14.8%. As a result, one batch of 46 batches of rice was screened out for AFB1 and AFB2, with a screening rate of 2.2%, of which the measured values were 10.8 μg/kg and 1.2 μg/kg, respectively. According to GB 2761-2017, the maximum allowable level of AFB1 in rice is 10 μg/kg; thus, the exceeding rate for AFB1 is 2.2%. Deoxynivalenol (DON) was screened out in 9 out of 24 batches of wheat (screening rate, 37.5%), while ZEN was screened out in 19 batches (screening rate, 79.2%). According to GB 2761-2017, the maximum allowable levels of DON and ZEN in wheat are 1000 and 60 μg/kg, respectively. The levels of DON and ZEN detected in the wheat samples did not exceed these limits. The proposed method uses MSE for qualitative screening to avoid the occurrence of false positives caused by interfering compounds with mass numbers and retention times similar to those of the analytes. TOF-MRM mode is then used to quantify the positively screened mycotoxins. The method is fast, accurate, sensitive, and suitable for the isolation and quantitative detection of mycotoxin residues in rice and wheat samples. The findings provide powerful technical support for mycotoxin contamination monitoring in rice and wheat and early risk-warning efforts.

Metabolic Rewiring in Tea Plants in Response to Gray Blight Disease Unveiled by Multi-Omics Analysis.

Gray blight disease, which is caused by Pestalotiopsis-like species, poses significant challenges to global tea production. However, the comprehensive metabolic responses of tea plants during gray blight infection remain understudied. Here, we employed a multi-omics strategy to characterize the temporal transcriptomic and metabolomic changes in tea plants during infection by Pseudopestalotiopsis theae, the causal agent of gray blight. Untargeted metabolomic profiling with ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOFMS) revealed extensive metabolic rewiring over the course of infection, particularly within 24 h post-inoculation. A total of 64 differentially accumulated metabolites were identified, including elevated levels of antimicrobial compounds such as caffeine and (-)-epigallocatechin 3-gallate, as well as oxidative catechin polymers like theaflavins, theasinensins and theacitrins. Conversely, the synthesis of (+)-catechin, (-)-epicatechin, oligomeric proanthocyanidins and flavonol glycosides decreased. Integrated omics analyses uncovered up-regulation of phenylpropanoid, flavonoid, lignin biosynthesis and down-regulation of photosynthesis in response to the pathogen stress. This study provides novel insights into the defense strategies of tea plants against gray blight disease, offering potential targets for disease control and crop improvement.

Fecal metabolomics combined with metagenomics sequencing to analyze the antidepressant mechanism of Yueju Wan

BackgroundYueju Wan (YJW), defined in Danxi's Mastery of Medicine, has Qi-regulating and Qi-promoting effects. YJW has frequently been applied in the clinic for the treatment of depression. Substantial evidence has shown that depression is related to metabolic abnormalities of the gut microbiota, and traditional Chinese medicine (TCM) can treat depression by adjusting gut microbiota metabolism. The antidepressant effect of YJW is well established, but thus far, whether its mechanism of action is achieved by regulating the intestinal flora has not been elucidated. MethodsIn this study, chronic unpredictable mild stress (CUMS) along with isolated feeding created a rat depression model, and YJW was administered for intervention. Rats were put through behavioral tests to determine their level of depression, and ELISA was utilized for measuring the level of monoamine neurotransmitters (MNTs) in the hippocampus. Metagenomic gene sequencing analysis was used to study the effect of depression on the intestinal flora in rats and the regulatory mechanism of YJW on the intestinal flora. Furthermore, ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q/TOF-MS) was utilized for fecal metabolomics studies to further reveal the antidepressant mechanism of YJW. The antidepressant mechanism of YJW was explored and further verified by Western blot analysis. ResultsDifferent doses of YJW improved the depressive state of rats and raised the levels of MNTs in the hippocampus. The results of metagenomic sequencing indicated that the YJW recovered the structure and diversity of the intestinal flora in depressed rats. Metabolomics revealed sustained changes in 21 metabolites after the treatment of YJW, suggesting that YJW can play an antidepressant role by improving abnormal metabolic pathways. The results of correlation analysis suggested that YJW might mediate Eubacterium, Oscillibacter, Roseburia, Romboutsia and Bacterium to regulate purine metabolism, tryptophan metabolism, primary bile acid biosynthesis, and glutamate metabolism and exert antidepressant effects. Western blot analysis showed that YJW reduced the content of IL-1β in the hippocampus, inhibited the activation of the NLRP3 inflammasome in the hippocampus of rats, and increased the content of ZO-1 in the colon of rats. ConclusionYJW can alleviate depressive symptoms in depressed rats, and its mechanism is connected to improving intestinal flora and regulating body metabolism.

Systematic Characterization of the Chemical Components and Absorbed Prototypes in Sanqi Xueshangning Capsules Using UPLC-Q-TOF-MS with SCIEX OS Software

Background Sanqi Xueshangning Capsules (SXC) is a famous Chinese patent medicine with a good curative effect on various gynecological pain diseases in China. However, its chemical composition and potential active ingredients were clearly unrevealed, which hindered the elaboration of its material basis functional mechanism and clinical application. Materials and Methods In the study, we developed an integrative ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) method coupled with SCIEX OS software for characterizing the chemical composition in SXC. The data collection was conducted using a Waters UPLCTM HSS T3 (2.1 × 100 mm, 1.8 µm) in a binary elution system. The SCIEX OS software coupled with multivariable processing techniques was used to screen absorbed prototypes in serum after oral administration of SXC. The structural identification of the components was performed by MS/MS fragmentation mechanism. Results A total of 113 components from SXC were tentatively characterized, including 38 saponins, 13 flavonoids, 10 phenylpropanoids, 9 alkaloids, 20 organic acids, and 23 others. Based on the in vitro composition results, 21 absorbed prototypes in rat serum were identified, mainly including alkaloids, flavonoids, phenylpropanoids, and saponins. Conclusion It was concluded that an integrative UPLC-Q-TOF-MS method coupled with SCIEX OS software could be successfully applied for the components’ characterization of SXC in vitro and in vivo, which laid a reliable scientific basis for promoting the discovery of material basis and the improvement of quality control of SXC.

Spatial metabolomics method to reveal differential metabolomes in microregions of Panax quinquefolius roots by using ultra-performance liquid chromatography quadrupole/time of flight-mass spectrometry and desorption electrospray ionization mass spectrometry imaging

Panax quinquefolius is a natural homology medicine and food that is rich in bioactive ingredients, such as ginsenosides and polysaccharides. The combination of ultra-performance liquid chromatography quadrupole/time of flight-mass spectrometry (UPLC-Q-TOF/MS) and desorption electrospray ionization mass spectrometry imaging (DESI-MSI) was used for the first time in a spatial metabolomics analysis to comprehensively evaluate the differential components in different microregions of P. quinquefolius. UPLC-Q-TOF/MS and DESI-MSI combined with principal component analysis and orthogonal partial least squares-discriminant analysis were used to screen differential metabolites. UPLC-Q-TOF/MS and DESI-MSI screened 27 and 23 differential metabolites, respectively, among which 15 differential metabolites were identified by both methods. It was found that some components, such as ginsenoside Rg1 and malonyl-ginsenoside Rc, were mainly distributed in P of the transverse slice of P. quinquefolius roots, while ginsenoside Ro and malonyl-ginsenoside Rd were mainly distributed in C. The methods and results of this study could be used to understand the precise localization, biosynthesis, and biological functions of special metabolites in P. quinquefolius.

Experimental study on changes in metabolic mechanism of papillary thyroid carcinoma complicated with Hashimoto’s thyroiditis

BackgroundWhether the mechanism of thyroid papillary carcinoma (PTC) is the same in patients with a Hashimoto's thyroiditis (HT) background as compared with patients with a normal background remains a highly debated and controversial issue. In this study, we aimed to analyze the differences and similarities of the metabolic mechanism of PTC in normal and HT background, and to explore the relationship between HT and PTC. MethodsThe ultra performance liquid chromatography-quadrupole-time of flight-mass spectrometry (UPLC-Q-TOF/MS) technology was used to analyze 61 PTC patient tissues (31 HT background and 30 normal tissue (NC) background). Potential biomarkers were screened from principal component analysis (PCA) to orthogonal partial least square (OPLS) discriminant analysis. HMDB was searched to identify potential differential metabolites and final metabolic pathway analysis was performed by MetaboAnalyst 5.0. We analyzed the differential metabolites diagnostic accuracy through receiver operating characteristic (ROC) curves analysis. ResultsSeven different metabolites were screened from HT group and NC group, including arginine, glutamic acid, cysteine, citric acid, malic acid, uracil and taurine. Logistic regression model combined with ROC analysis of these 7 biomarkers had good discriminability for PTC (area under operating characteristic curve of HT group and NC group were 0.867 and 0.973, respectively). The HT group had specific metabolic pathways, including aminoacyl-tRNA biosynthesis, glycine, serine and threonine metabolism. ConclusionsThe metabolic profiles of the NC and HT groups had important similarities and differences in PTC. The correlation of PTC with HT may be related to aminoacyl-tRNA biosynthesis, serine and threonine metabolism.

Chemical Composition Analysis of Lilium brownii var. Viridulum Baker and the Effect of Postharvest Primary Processing on Its Quality

To elucidate ths chemical composition of Lilium brownii var. Viridulum Baker and the impact of primary processing on its quality, a comparison was made on the polysaccharide and extract contents of ten batches of Lilium from different regions, including Lilium lancifolium Thunb., Lilium brownii var. Viridulum Baker, and Lilium davidi var. unicdor cotton. The chemical composition differences of the three Lilium species mentioned above were compared using high-performance liquid chromatography (HPLC) fingerprinting. The chemical components of Lilium brownii var. Viridulum Baker were analyzed by ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MSE). The boiling time and drying temperature of fresh Lilium brownii var. Viridulum Baker were investigated using the comprehensive scoring method based on the polysaccharide and extract contents. The results showed that the polysaccharide content of ten batches of Lilium from different origins ranged from 13.34% to 34.00%, and the extract content ranged from 27.10% to 47.10%. The HPLC fingerprinting results showed that the similarity of the three Lilium species ranged from 0.796 to 0.999, and the chemical components of Lilium davidi var. unicdor cotton differed significantly from those of Lilium lancifolium Thunb. and Lilium brownii var. Viridulum Baker. UPLC-Q-TOF-MSE identified 22 phenolic compounds, 35 steroidal saponins, and 6 alkaloids in Lilium brownii var. Viridulum Baker. Boiling for 5 min and drying at 85 °C has the least effect on the quality of fresh Lilium.

UPLC-Q-TOF-MS, network analysis, and molecular docking to investigate the effect and active ingredients of tea-seed oil against bacterial pathogens.

Object: This research intended to probe the antibacterial effect and pharmacodynamic substances of Tea-Seed Oil (TSO) through the use of ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) analysis, network analysis, and molecular docking. Methods: The major chemical components in the methanol-extracted fractions of TSO were subjected to UPLC-Q-TOF-MS. Network pharmacology and molecular docking techniques were integrated to investigate the core components, targets, and potential mechanisms of action through which the TSO exert their antibacterial properties. To evaluate the inhibitory effects, the minimum inhibitory concentration and diameter of the bacteriostatic circle were calculated for the potential active ingredients and their equal ratios of combinatorial components (ERCC) against Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans. Moreover, the quantification of the active constituents within TSO was achieved through the utilization of high-performance liquid chromatography (HPLC). Results: The methanol-extracted fractions contained a total of 47 chemical components, predominantly consisting of unsaturated fatty acids and phenolic compounds. The network pharmacology analysis and molecular docking analysis revealed that various components, including gallocatechin, gallic acid, epigallocatechin, theophylline, chlorogenic acid, puerarin, and phlorizin, have the ability to interact with critical core targets such as serine/threonine protein kinase 1 (AKT1), epidermal growth factor receptor (EGFR), a monoclonal antibody to mitogen-activated protein kinase 14 (MAPK14), HSP90AA1, and estrogen receptor 1 (ESR1). Furthermore, these components can modulate the phosphatidylinositol-3-kinase protein kinase B (PI3K-AKT), estrogen, MAPK and interleukin 17 (IL-17) signaling pathways, hereby exerting antibacterial effects. In vitro validation trials have found that seven components, namely gallocatechin, gallic acid, epigallocatechin, theophylline, chlorogenic acid, puerarin, and phloretin, displayed substantial inhibitory effects on E. coli, S. aureus, P. aeruginosa, and C. albicans, and are typically present in tea oil, with a total content ranging from 15.87∼24.91 μg·g-1. Conclusion: The outcomes of this investigation possess the possibility to expand our knowledge base concerning the utilization of TSO, furnish a theoretical framework for the exploration of antibacterial drugs and cosmetics derived from inherently occurring TSO, and establish a robust groundwork for the advancement and implementations of TOS products within clinical settings.

Pharmacokinetics, anti-rheumatoid arthritis activity, and active ingredient contents of Eucommia ulmoides Oliv

Eucommia ulmoides Oliv. is a deciduous tree which contains various chemical ingredients. The main objective was to document the active chemical ingredients of Eucommia ulmoides Oliv. and their metabolic profiles in vivo, with a view to providing an experimental and theoretical basis for clarifying the mechanisms underlying the pharmacological activity of Eucommia ulmoides Oliv. against rheumatoid arthritis. Eight main active constituents of Eucommia ulmoides Oliv. bark (pinoresinol glucopyranoside, aucubin, geniposidic acid, geniposide, genipin, chlorogenic acid, quercetin and betulinic acid) were quantified using high-performance liquid chromatography (HPLC). This paper additionally identified and characterized prototype metabolites via ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) combined with the Human Metabolome Database (HMDB) and literature comparisons. Ultra pressure liquid chromatography-mass spectrometry/ mass spectrometry (UPLC-MS/MS) was subsequently employed to quantify these components in blood over time and evaluate their pharmacokinetic characteristics. The anti-rheumatoid arthritis effects of genipin, pinoresinol glucopyranoside and their combinations were assessed using in vitro cellular assays. We identified and characterized a total of 53 ingredients from Eucommia ulmoides Oliv. bark and plasma samples, among which 20 were confirmed as prototype metabolites. Meanwhile, this paper derived and analyzed the metabolic cleavage pathway of 8 index ingredients. Six of these compounds displayed rapid entry into blood, with high plasma exposure and fast elimination rates. Data from the in vitro cellular assay showed that aucubin, pinoresinol glucopyranoside, genipin, and combinations of these compounds effectively inhibit MH7A cell proliferation, reduce NO release, and decrease inflammatory factor levels.

Protective effect and mechanism of styrax on ischemic stroke rats: metabonomic insights by UPLC-Q/TOF-MS analysis

Context Styrax is used for prevention and treatment of cerebrovascular diseases. However, the underlying mechanism remains unclear. Objective To elucidate styrax’s anti-ischemic stroke protective effects and underlying mechanisms. Materials and methods An ischemic-stroke rat model was established based on middle cerebral artery occlusion (MCAO). Sprague-Dawley rats were randomly assigned to the following groups (n = 10) and administered intragastrically once a day for 7 consecutive days: sham, model, nimodipine (24 mg/kg), styrax-L (0.1 g/kg), styrax-M (0.2 g/kg) and styrax-H (0.4 g/kg). Neurological function, biochemical assessment, and ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UPLC-Q/TOF-MS)-based serum metabonomics were used to elucidate styrax’s cerebral protective effects and mechanisms. Pearson correlation and western blot analyses were performed to verify. Results The addition of 0.4 g/kg styrax significantly reduced cerebral infarct volume and neurobehavioral abnormality score. Different doses of styrax also decrease MDA, TNF-α, IL-6, and IL-1β, and increase SOD and GSH-Px in ischemic-stroke rats (p < 0.05; MDA, p < 0.05 only at 0.4 g/kg dose). Biochemical indicators and metabolic-profile analyses (PCA, PLS-DA, and OPLS-DA) also supported styrax’s protective effects. Endogenous metabolites (22) were identified in ischemic-stroke rats, and these perturbations were reversible via styrax intervention, which is predominantly involved in energy metabolism, glutathione and glutamine metabolism, and other metabolic processes. Additionally, styrax significantly upregulated phosphorylated AMP-activated protein kinase and glutaminase brain-tissue expression. Conclusion Styrax treatment could ameliorate ischemic-stroke rats by intervening with energy metabolism and glutamine metabolism. This can help us understand the mechanism of styrax, inspiring more clinical application and promotion.

Mechanisms of Xiaozheng decoction for anti-bladder cancer effects via affecting the GSK3β/β-catenin signaling pathways: a network pharmacology-directed experimental investigation

PurposeThe combination of Xiaozheng decoction with postoperative intravesical instillation has been shown to improve the prognosis of bladder cancer patients and prevent recurrence. However, the mechanisms underlying the efficacy of this herbal formula remain largely unclear. This research aims to identify the important components of Xiaozheng decoction and explore their anti-bladder cancer effect and mechanism using network pharmacology-based experiments.MethodsThe chemical ingredients of each herb in the Xiaozheng decoction were collected from the Traditional Chinese Medicine (TCM) database. Network pharmacology was employed to predict the target proteins and pathways of action. Disease databases were utilized to identify target genes associated with bladder cancer. A Protein–Protein Interaction (PPI) network was constructed to illustrate the interaction with intersected target proteins. Key targets were identified using Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) functional enrichment analysis. A compound-target-pathway network was established after molecular docking predictions. In vitro experiments with bladder cancer cell lines were conducted using core chemical components confirmed by ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-qTOF-MS) to verify the conclusions of network pharmacology.Results45 active compounds were extracted, and their relationships with Traditional Chinese Medicines (TCMs) and protein targets were presented, comprising 7 herbs, 45 active compounds, and 557 protein targets. The intersection between potential TCM target genes and bladder cancer-related genes yielded 322 genes. GO and KEGG analyses indicated that these targets may be involved in numerous cancer-related pathways. Molecular docking results showed that candidate compounds except mandenol could form stable conformations with the receptor. In vitro experiments on three bladder cancer cell lines demonstrated that quercetin and two other impressive new compounds, bisdemethoxycurcumin (BDMC) and kumatakenin, significantly promoted cancer cell apoptosis through the B-cell lymphoma 2/Bcl-2-associated X (Bcl-2/BAX) pathway and inhibited proliferation and migration through the glycogen synthase kinase 3 beta (GSK3β)/β-catenin pathway.ConclusionBy employing network pharmacology and conducting in vitro experiments, the mechanism of Xiaozheng decoction’s effect against bladder cancer was tentatively elucidated, and its main active ingredients and targets were identified, providing a scientific basis for future research.