Jiawei Kongsheng Zhenzhong Pill: marker compounds, absorption into the serum (rat), and Q-markers identified by UPLC-Q-TOF-MS/MS.

Background: The Jiawei Kongsheng Zhenzhong pill (JKZP), a Chinese herbal prescription comprised of eight Chinese crude drugs, has been historically employed to treat neurological and psychological disorders. Nevertheless, the ambiguous material basis severely hindered its progress and application. Purpose: The current study aimed to establish a rapid analytical method for identifying the chemical components of the JKZP aqueous extract and the components absorbed into the rat serum to investigate the quality markers (Q-markers) responsible for the neuroprotective effects of JKZP. Methods: The qualitative detection of the chemical components, prototype components, and metabolites of the aqueous extracts of JKZP, as well as the serum samples of rats that were administered the drug, was performed using the ultra-performance liquid chromatography- quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS) technology. This analysis combined information from literature reports and database comparisons. Moreover, the study was conducted to anticipate the potential Q-markers for the neuroprotective effects of JKZP based on the "five principles" of Q-marker determination. Results: A total of 67 compounds and 111 serum components (comprising 33 prototypes and 78 metabolites) were detected and identified. Combining the principles of quality transmission and traceability, compound compatibility environment, component specificity, effectiveness, and measurability, the study predicted that five key compounds, namely, senkyunolide H, danshensu, echinacoside, loganin, and 3,6'-disinapoyl sucrose, may serve as potential pharmacological bases for the neuroprotective effects of JKZP. Conclusion: To summarize, the UPLC-Q-TOF-MS/MS technique can be employed to rapidly and accurately identify compounds in JKZP. Five active compounds have been predicted to be the Q-markers for the neuroprotective effects of JKZP. This discovery serves as a reference for improving quality, advancing further research and development, and utilizing Chinese herbal prescriptions.

Exploration the mechanism of Shenling Baizhu San in the treatment of chronic obstructive pulmonary disease based on UPLC-Q-TOF-MS/MS, network pharmacology and in vitro experimental verification.

ETHNOPHARMACOLOGICAL RELEVANCE: Shenling Baizhu San (SLBZS) is a formula of traditional Chinese medicine (TCM) that enhances the functions of the qi, spleen, and lung. According to the theory of TCM, chronic obstructive pulmonary disease (COPD) is often caused by lung qi deficiency, and SLBZS is often used in the treatment of COPD and has achieved remarkable results. However, the active components of SLBZS absorbed in serum and the underlying mechanism of SLBZS in treating COPD remain unclear and require further studies. AIM OF THE STUDY: The objective of this study is to investigate the active components of SLBZS in rat serum, as well as the crucial targets and signaling pathways involved in the therapeutic effects of SLBZS for COPD. MATERIALS AND METHODS: First, the absorption components and metabolites of SLBZS in rat serum were identified using ultra-performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS). Second, potential targets of SLBZS for the treatment of COPD were acquired from publicly accessible online sources. Cytoscape (v3.7.0) software was used to construct a component-target-pathway network and a protein-protein interaction (PPI) network. The Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis of potential targets was performed using the Metascape database. The binding status of the active components in SLBZS to the potential targets was assessed with molecular docking technology. Finally, a cell model of COPD was successfully developed for experimental validation In vitro. RESULTS: A total of 108 active components were identified, including 30 prototype components and 78 metabolites. A total of 292 potential targets for the treatment of COPD were identified, including TNF, IL-6, TLR9, RELA, and others. The KEGG pathway included inflammatory mediator regulation of TRP channels, necroptosis, and the NF-κB signaling pathway, among others. The In vitro experiments showed that SLBZS-containing serum had the ability to decrease the levels of inflammatory factors and cell death. Additionally, it was observed that SLBZS-containing serum could control the expression levels of TLR9, MyD88, TRAF6, NF-κB, and IκBα at the mRNA and protein levels. These findings suggested that SLBZS-containing serum was likely to be involved in the regulation of the TLR9/NF-κB pathway. CONCLUSIONS: The mechanism of action of SLBZS on COPD was preliminarily elucidated using UPLC-Q-TOF-MS/MS, network pharmacology, and In vitro experiments. The primary active components and potential targets of SLBZS were identified, providing a scientific foundation for further research.

Melatonin attenuates chronic intermittent hypoxia-induced intestinal barrier dysfunction in mice.

BACKGROUND AND PURPOSE: Chronic intermittent hypoxia (CIH) triggers subclinical intestinal barrier disruption prior to systemic low-grade inflammation. Increasing evidence suggests therapeutic effects of melatonin on systemic inflammation and gut microbiota remodelling. However, whether and how melatonin alleviates CIH-induced intestinal barrier dysfunction remains unclear. EXPERIMENTAL APPROACH: C57BL/6 J mice and Caco-2 cell line were treated. We evaluated gut barrier function spectrophotometrically using fluorescein isothiocyanate (FITC)-labelled dextran. Immunohistochemical and immunofluorescent staining were used to detect morphological changes in the mechanical barrier. Western blotting (WB) and quantitative real-time polymerase chain reaction (qRT-PCR) revealed the expression of tight junctions, signal transducer and activator of transcription 3 (STAT3) levels. 16 S rRNA analysis of the colonic contents microflora. Flow cytometry was used to detect cytokines and Th17 cells with and without melatonin supplementation. KEY RESULTS: We found that CIH could induce colonic mucosal injury, including reduction in the number of goblet cells and decrease the expression of intestinal tight junction proteins. CIH could decrease the abundance of the beneficial genera Clostridium, Akkermansia, and Bacteroides, while increasing the abundance of the pathogenic genera Desulfovibrio and Bifidobacterium. Finally, CIH facilitated Th17 differentiation via the phosphorylation of signal transducer and activator of transcription 3 (STAT3) in vitro and elevated the circulating pro-inflammatory cytokine in vivo. Melatonin supplementation ameliorated CIH-induced intestinal mucosal injury, gut microbiota dysbiosis, enteric Th17 polarization, and systemic low-grade inflammation reactions mentioned-above. CONCLUSION AND IMPLICATIONS: Melatonin attenuated CIH-induced intestinal barrier dysfunction by regulating gut flora dysbiosis, mucosal epithelium integrity, and Th17 polarization via STAT3 signalling.

Effects of Ilicis Chinensis folium extract supplementation on growth performance, serum parameters, intestinal morphology, and antioxidant capacity of broiler chickens.

BACKGROUND: Ilicis chinensis folium extract (ICFE) is a powder extracted and processed with Ilex chinensis Sims (ICS) which has numerous bioactivities and is conventionally used in traditional Chinese medicine. Nonetheless, there has been no definitive study evaluating ICFE's application as a feed supplement for broilers. This research sought to determine the chemical composition and evaluate how dietary ICFE supplementation affects the growth performance, serum metrics, intestinal structure, and antioxidant capacity of broilers. METHODS: A total of 360 one-day-old broiler chicks were assigned to four treatments (with 9 replicates of 10 chicks, each) of dietary supplementation with ICFE at 0, 250, 500, and 1,000 mg /kg for 42 days. RESULTS: Ten polyphenolic compounds and two triterpenoid glycosides were detected by HPLC. In the grower stage and overall, broilers supplemented with 500 and 1,000 mg/kg ICFE exhibited a higher ADFI (P < 0.05) than the controls. Additionally, compared to the controls, broilers receiving low, medium, or high dosages of ICFE exhibited higher average daily gains (P < 0.05) throughout the starter stage and overall. Organ indices showed no significant variation, suggesting that ICFE was non-toxic. ICFE supplementation increased the height of villi in the duodenum and jejunum, reduced crypt depth, and increased the villus/crypt ratio in the duodenum (P < 0.05). Serum concentrations of IL-4 and IgA were increased in ICFE-supplemented broilers. The serum malondialdehyde concentration was reduced, whereas superoxide dismutase activity and total antioxidant capacity increased through supplementation with ICFE. CONCLUSION: ICFE supplementation can improve intestinal morphology, antioxidant capacity, and growth performance of broilers. Hence, ICFE is a promising and safe alternative to antibiotics in broilers, and 500 mg/kg appears to be the optimal dose.

Encapsulation of propionic acid within the aqueous phase of water-in-oil emulsions: reduced thermal volatilization and enhanced gastrointestinal stability.

Propionic acid (PA) is a water-soluble substance that has been shown to be beneficial for improving colon-related diseases. However, its appliance as a nutraceutical ingredient is hampered by its volatility, irritating odor, and easy absorption in the stomach and small intestine. A chitosan solution containing propionic acid was dispersed in a palm oil/corn oil mixture with polyglycerol polyricinoleate (PGPR) to form PA-loaded water-in-oil (W/O) emulsions. The stability of the emulsions was improved by the inclusion of both chitosan and palm oil, where the chitosan reduced the emulsion particle size and palm oil increased the viscosity. The thermal volatility and storage stability of the encapsulated propionic acid were significantly improved due to the stability of emulsion structure and hydrogen bonding between chitosan and propionic acid. Around 56% of propionic acid remained within the aqueous phase after the simulated gastrointestinal digestion. Our results indicate that W/O emulsions might be candidates as colon-targeted delivery systems for propionic acid, which could be beneficial for maintaining colon health.

Identification and validation of metabolism-related genes signature and immune infiltration landscape of rheumatoid arthritis based on machine learning.

Rheumatoid arthritis (RA) causes irreversible joint damage, but the pathogenesis is unknown. Therefore, it is crucial to identify diagnostic biomarkers of RA metabolism-related genes (MRGs). This study obtained transcriptome data from healthy individuals (HC) and RA patients from the GEO database. Weighted gene correlation network analysis (WGCNA), the least absolute shrinkage and selection operator (LASSO), and random forest (RF) algorithms were adopted to identify the diagnostic feature biomarker for RA. In addition, biomarkers were verified by qRT-PCR and Western blot analysis. We established a mouse model of collagen-induced arthritis (CIA), which was confirmed by HE staining and bone structure micro-CT analysis, and then further verified the biomarkers by immunofluorescence. In vitro NMR analysis was used to analyze and identify possible metabolites. The correlation of diagnostic feature biomarkers and immune cells was performed using the Spearman-rank correlation algorithm. In this study, a total of 434 DE-MRGs were identified. GO and KEGG enrichment analysis indicated that the DE-MRGs were significantly enriched in small molecules, catabolic process, purine metabolism, carbon metabolism, and inositol phosphate metabolism. AKR1C3, MCEE, POLE4, and PFKM were identified through WGCNA, LASSO, and RF algorithms. The nomogram result should have a significant diagnostic capacity of four biomarkers in RA. Immune infiltration landscape analysis revealed a significant difference in immune cells between HC and RA groups. Our findings suggest that AKR1C3, MCEE, POLE4, and PFKM were identified as potential diagnostic feature biomarkers associated with RA's immune cell infiltrations, providing a new perspective for future research and clinical management of RA.

Capsaicin encapsulated in W/O/W double emulsions fabricated via ethanol-induced pectin gelling: Improvement of bioaccessibility and reduction of irritation.

Capsaicin is a water-insoluble bioactive component with several beneficial physiological functions. However, the widespread application of this hydrophobic phytochemical is limited by its low water-solubility, intense irritation, and poor bioaccessibility. These challenges can be overcome by entrapping capsaicin within the internal water phase of water-in-oil-in-water (W/O/W) double emulsions via using ethanol to induce pectin gelling. In this study, ethanol was used both to dissolve capsaicin and to promote pectin gelation, thereby forming capsaicin-loaded pectin hydrogels that were used as the internal water phase of the double emulsions. Pectin addition improved the physical stability of the emulsions and led to a high encapsulation efficiency of capsaicin (>70 % after 7d storage). After simulated oral and gastric digestion, capsaicin-loaded double emulsions maintained their compartmentalized structure, avoiding capsaicin leakage in the month and stomach. The double emulsions were digested in the small intestine, thereby releasing the capsaicin. Capsaicin bioaccessibility was significantly enhanced after encapsulation, which was attributed to mixed micelle formation by the digested lipid phase. Furthermore, encapsulation of capsaicin within the double emulsions reduced the irritation in the gastrointestinal tissues of mice. This kind of double emulsion may have great potential for the development of more palatable capsaicin-loaded functional food products.

CT-determined sarcopenia is associated with neutropenia in patients undergoing hyperthermic intraperitoneal chemotherapy for gastrointestinal cancer.

BACKGROUND: With better patient selection and the increasing experience in patients undergoing hyperthermic intraperitoneal chemotherapy (HIPEC) combined surgery, the rate of severe postoperative complications and mortality decreased significantly. However, leukopenia and neutropenia were still a particular concern, and their relation to sarcopenia was not clarified. METHODS: Data of consecutive patients who underwent HIPEC for gastrointestinal cancer were collected and analyzed retrospectively between September 2020 and August 2022. Sarcopenia was assessed using psoas muscle index (PMI) at the L3 level on preoperative computed tomography (CT). RESULTS: Among 103 patients enrolled, 37 (35.9%) were classified as sarcopenic. Most leukopenia and neutropenia occurred during the hospital leaving period after HIPEC and surgery. Before the first time of postoperative chemotherapy, the blood tests revealed 11 (29.73%) and 6 (9.09%) patients were diagnosed with neutropenia in sarcopenia and no sarcopenia groups, respectively. Logistic regression analysis revealed sarcopenia was independently associated with the increased risk of neutropenia (OR 5.58, 95% CI 1.70-18.29, p = 0.005). An incremental albumin level was protective against the occurrence of leukopenia and neutropenia. CONCLUSIONS: Sarcopenia and low albumin level were significantly associated with an increased rate of delayed neutropenia after HIPEC in that disease setting and could be the preoperative risk predictors.

Neuroprotective effects of salidroside on ageing hippocampal neurons and naturally ageing mice via the PI3K/Akt/TERT pathway.

Studies have found that salidroside, isolated from Rhodiola rosea L, has various pharmacological activities, but there have been no studies on the effects of salidroside on brain hippocampal senescence. The purpose of this study was to investigate the mechanistic role of salidroside in hippocampal neuron senescence and injury. In this study, long-term cultured primary rat hippocampal neurons and naturally aged C57 mice were treated with salidroside. The results showed that salidroside increased the viability and MAP2 expression, reduced ß-galactosidase (ß-gal) levels of rat primary hippocampal neurons. Salidroside also improved cognition dysfunction in ageing mice and alleviated neuronal degeneration in the ageing mice CA1 region. Moreover, salidroside decreased the levels of oxidative stress and p21, p16 protein expressions of hippocampal neurons and ageing mice. Salidroside promoted telomerase reverse transcriptase (TERT) protein expression via the phosphatidylinositol-3-kinase (PI3K)/protein kinase B (Akt) pathway. In conclusion, our findings suggest that salidroside has the potential to be used as a therapeutic strategy for anti-ageing and ageing-related disease treatment.

Enhancing the oxidative stability of algal oil emulsions by adding sweet orange oil: Effect of essential oil concentration.

The effects of sweet orange essential oil (SOEO) concentration (0-12.5% of oil phase) on the physical stability, oxidative stability, and interfacial composition of algal oil-in-water emulsions containing sodium caseinate-coated oil droplets was examined. SOEO addition had no influence on the microstructure and physical stability of the algal oil emulsions. The addition of SOEO enhanced the oxidation stability of algal oil emulsion. As an example, the values of algal oil emulsions with 0 and 10% SOEO were 198 and 100 mmol/kg algal oil after 16 days of accelerated oxidation, respectively. The absorbed protein level was higher in the algal oil emulsion containing 10% SOEO (70%) than in 0% SOEO (57%). This result suggested that the presence of SOEO enhanced the interfacial thickness, possibly by interacting with the casein molecules. A thicker protein layer may have helped to retard the oxidation of the omega-3 oils inside lipid droplets.

A rapid and universal method for isolating starch granules in plant tissues.

Starch is the major form of carbohydrate storage in plants and exists as discrete starch granules (SGs). Isolation of high-quality SGs in different plant tissues is a prerequisite for studying the roles of SGs during plant growth, development, and responses to abiotic stress. However, it is difficult to isolate transitory SGs from leaves and storage SGs from pollen grains due to their small sizes and low quantities. Herein, we develop a novel method for isolating SGs by using the aqueous two-phase system (ATS) of ethanol/NaH2 PO4 . The ATS method efficiently separated SGs from contaminants based on their differences in density, solubility, and polarity. Using this method, we first isolated and purified three kinds of SGs from maize seeds, pollen, and leaves. The biochemical, microscopic, and proteomic analyses demonstrated the high purity of the isolated SGs. Proteomic analysis revealed distinct differences in SG-bound proteins between seed SGs and pollen SGs. As a simple, rapid, and low-cost method, the ATS-based method exhibits highly universal and reproducible results for starch-containing tissues in various plant species.

Clonal expansion of SIV-infected cells in macaques on antiretroviral therapy is similar to that of HIV-infected cells in humans.

Clonal expansion of HIV infected cells plays an important role in the formation and persistence of the reservoir that allows the virus to persist, in DNA form, despite effective antiretroviral therapy. We used integration site analysis to ask if there is a similar clonal expansion of SIV infected cells in macaques. We show that the distribution of HIV and SIV integration sites in vitro is similar and that both viruses preferentially integrate in many of the same genes. We obtained approximately 8000 integration sites from blood samples taken from SIV-infected macaques prior to the initiation of ART, and from blood, spleen, and lymph node samples taken at necropsy. Seven clones were identified in the pre-ART samples; one persisted for a year on ART. An additional 100 clones were found only in on-ART samples; a number of these clones were found in more than one tissue. The timing and extent of clonal expansion of SIV-infected cells in macaques and HIV-infected cells in humans is quite similar. This suggests that SIV-infected macaques represent a useful model of the clonal expansion of HIV infected cells in humans that can be used to evaluate strategies intended to control or eradicate the viral reservoir.

Efficacy and Safety of Vitamin D Supplementation in Patients With Systemic Lupus Erythematosus: A Meta-analysis of Randomized Controlled Trials.

BACKGROUND: The efficacy of vitamin D supplementation in patients with systemic lupus erythematosus (SLE) remains uncertain. This meta-analysis aimed to systematically evaluate the efficacy and safety of vitamin D supplementation in patients with SLE. MATERIALS AND METHODS: Randomized controlled trials (RCTs) were searched in PubMed, Embase, Cochrane CENTRAL and Web of Science databases. The retrieved studies were subjected to meta-analysis using the fixed-effect or random-effect model. RESULTS: Five eligible RCTs enrolling 490 participants were included. Compared to the placebo treatment, vitamin D supplementation significantly increased the level of serum 25-hydroxyvitamin D (25(OH)D) (5 trials, 490 participants: standard mean difference (SMD) = 2.072, 95% CI: 1.078-3.066, P < 0.001). The pooled result from 2 RCTs showed that vitamin D supplementation decreased the fatigue severity scale scores in patients with SLE (2 trials, 79 participants: SMD = -1.179, 95% CI: -1.897 to -0.460, P = 0.001). The SLE disease activity index scores and positivity of anti-double-stranded DNA antibodies (anti-dsDNA) did not differ significantly (4 trials, 223 participants: SMD = -0.507, 95% CI: -1.055-0.041, P = 0.070; 3 trials, 361 participants: Risk ratio = 0.880, 95% CI: 0.734-1.054, P = 0.165) between the vitamin D supplementation group and the placebo treatment group. None of the included studies reported severe adverse events associated with vitamin D supplementation. CONCLUSIONS: This meta-analysis suggested that vitamin D supplementation is effective in increasing the serum 25(OH)D levels, may improve fatigue, and is well-tolerated in patients with SLE, however, it does not seem to have significant effects in decreasing the positivity of anti-dsDNA and disease activity.

Protective effects of a Ganoderma atrum polysaccharide against acrylamide induced oxidative damage via a mitochondria mediated intrinsic apoptotic pathway in IEC-6 cells.

The preventive role of a purified Ganoderma atrum polysaccharide PSG-1-F2 as a new dietary antioxidant against the intestinal toxicity of acrylamide (ACR) was investigated in vitro. Our results showed that ACR could induce oxidative stress in IEC-6 cells by the overproduction of reactive oxygen species (ROS) and malondialdehyde (MDA), and as well as the reduction in the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px). In addition, the induction of a mitochondria-mediated intrinsic apoptotic pathway by ACR was evidenced by the events of loss of mitochondrial membrane potential, bax/bcl-2 dysregulation, cytochrome c release, and activation of caspase-3. Interestingly, PSG-1-F2 was able to suppress ACR toxicity by improving the redox status of IEC-6 cells and by attenuating mitochondria-mediated apoptosis. Its protective effect was even superior to the clinically used antioxidant N-acetylcysteine (NAC). This study uniquely introduces PSG-1-F2 as a potential inhibitor of ACR-induced stress and toxicities.

Combining near infrared spectroscopy with predictive model and expertise to monitor herb extraction processes.

Albeit extensively utilized, herb extraction process (HEP) is hard to be monitored because of its batch nature and the fluctuating quality of raw materials. Process analytical tools like near infrared spectroscopy (NIRS) can offer nondestructive examinations and collect abundant data of the process, which in principle contain the information about the quality of both the product and the process itself. However, extra effort is often required for the data mining of such process measurements, and extracting knowledge of the quality of process can be even harder. In this study, we take the extraction process of licorice as a typical HEP instance, and combine NIRS with classical partial least squared regression (PLSR) and expertise for its on-line monitoring. We show that our scheme effectively extracts information with clear physical meanings, through which we can even uncover the process fault that does not induce evident abnormalities in the product quality. Moreover, the constructed model can continuously evolve with more process data from daily operations, and the idea of the whole framework can be directly generalized to other HEP.

Near infrared spectroscopy based monitoring of extraction processes of raw material with the help of dynamic predictive modeling.

The control of batch-to-batch quality variations remains a challenging task for pharmaceutical industries, e.g., traditional Chinese medicine (TCM) manufacturing. One difficult problem is to produce pharmaceutical products with consistent quality from raw material of large quality variations. In this paper, an integrated methodology combining the near infrared spectroscopy (NIRS) and dynamic predictive modeling is developed for the monitoring and control of the batch extraction process of licorice. With the spectra data in hand, the initial state of the process is firstly estimated with a state-space model to construct a process monitoring strategy for the early detection of variations induced by the initial process inputs such as raw materials. Secondly, the quality property of the end product is predicted at the mid-course during the extraction process with a partial least squares (PLS) model. The batch-end-time (BET) is then adjusted accordingly to minimize the quality variations. In conclusion, our study shows that with the help of the dynamic predictive modeling, NIRS can offer the past and future information of the process, which enables more accurate monitoring and control of process performance and product quality.

1H NMR combined with chemometrics for the rapid detection of adulteration in camellia oils.

Proton nuclear magnetic resonance (1H NMR) and chemometrics were employed to detect the adulteration of camellia oil (CAO) with 3 different cheap vegetable oils. With the intensity of 15 selected 1H NMR signals as input variables, principal component analysis (PCA) showed good group clustering results for pure and nonpure CAO, but unsatisfied identification accuracy for the adulterated oil types, indicating relatively small difference among those oils. Whereas these difference could be revealed by orthogonal projection to latent structures discriminant analysis (OPLS-DA), with identification accuracy higher than 90%. Partial least squares (PLS) was further applied for the prediction of adulteration level in CAO. With less than 6 variables screened out by variable importance in the projection (VIP) scores as potential key markers, the developed PLS models showed better accuracy. The prediction results for 10 hold-out samples also confirmed that this method was accurate and fast for the detection of CAO adulteration.

Atractylodes lactone compounds inhibit platelet activation.

Platelets play a crucial role in the development and progression of atherosclerosis-thrombosis and, therefore, antiplatelet drugs are widely used in the treatment of coronary artery disease. Moreover, advances in understanding the biological functions of natural plant products can provide new pharmacological strategies aimed at promoting cardiovascular health. Atractylenolide I (ATL-1), ATL-2, and ATL-3 are the major bioactive components of a Qi tonifying medicinal herb Rhizoma Atractylodis Macrocephalae (Atractylodes macrocephala), which is commonly used in traditional Chinese medicine (TCM). These components possess well-documented anti-inflammatory and anticancer activities, but their effects on platelet activation are still unknown. In this study, the effects of ATL on platelet function in vitro and in vivo were investigated, and the underlying mechanism was explored. We found that ATL-2 and ATL-3 but not ATL-1 diminished agonist-induced platelet aggregation and diminished adenosine triphosphate (ATP) release from dense granules. The levels of phospho-Akt (Ser473) and phospho-p38 MAPK were downregulated in the presence of ATL-2 and ATL-3. We also found that ATL-2 and ATL-3 have a similar inhibitory effect on platelet activation as acetylsalicylic acid in response to agonists. Furthermore, ATL-2 and ATL-3 diminished the spreading of human platelets on immobilized fibrinogen (Fg), delayed clot retraction in platelet-depleted plasma containing human platelets, extended first occlusion time in a mouse model of ferric chloride (FeCl3)-induced carotid arterial thrombosis, and prolonged the bleeding time. These observations suggest that ATL-2 and ATL-3 are potential candidate therapeutic drugs for the prevention of thrombosis.

Proteomic Characterization of Differential Abundant Proteins Accumulated between Lower and Upper Epidermises of Fleshy Scales in Onion (Allium cepa L.) Bulbs.

The onion (Allium cepa L.) is widely planted worldwide as a valuable vegetable crop. The scales of an onion bulb are a modified type of leaf. The one-layer-cell epidermis of onion scales is commonly used as a model experimental material in botany and molecular biology. The lower epidermis (LE) and upper epidermis (UE) of onion scales display obvious differences in microscopic structure, cell differentiation and pigment synthesis; however, associated proteomic differences are unclear. LE and UE can be easily sampled as single-layer-cell tissues for comparative proteomic analysis. In this study, a proteomic approach based on 2-DE and mass spectrometry (MS) was applied to compare LE and UE of fleshy scales from yellow and red onions. We identified 47 differential abundant protein spots (representing 31 unique proteins) between LE and UE in red and yellow onions. These proteins are mainly involved in pigment synthesis, stress response, and cell division. Particularly, the differentially accumulated chalcone-flavanone isomerase and flavone O-methyltransferase 1-like in LE may result in the differences in the onion scale color between red and yellow onions. Moreover, stress-related proteins abundantly accumulated in both LE and UE. In addition, the differential accumulation of UDP-arabinopyranose mutase 1-like protein and ß-1,3-glucanase in the LE may be related to the different cell sizes between LE and UE of the two types of onion. The data derived from this study provides new insight into the differences in differentiation and developmental processes between onion epidermises. This study may also make a contribution to onion breeding, such as improving resistances and changing colors.

Antagonism of Ca2+ influx via L-type Ca2+ channels mediates the vasorelaxant effect of Catharanthus roseus-derived vindorosine in rat renal artery.

Catharanthus roseus is a traditional herbal medicine used in Asian and African countries for the treatment of various diseases including hypertension. The present study examined possible cellular mechanisms for the relaxation of rat renal arteries induced by vindorosine extracted from C. roseus. Intrarenal arteries were isolated from 200-300 g male Sprague-Dawley rats and treated with different pharmacological blockers and inhibitors for the measurement of vascular reactivity on a Multi Myograph System. Fluorescence imaging by laser scanning confocal microscopy was utilized to determine the intracellular Ca(2+) level in the vascular smooth muscles of the renal arteries. Vindorosine in micromolar concentrations relaxes renal arteries precontracted by KCl, phenylephrine, 11-dideoxy-9α,11α-epoxymethanoprostaglandin F2α, and serotonin. Vindorosine-induced relaxations were unaffected by endothelium denudation or by treatment with the nitric oxide synthase inhibitor N (G)-nitro-L-arginine methyl ester hydrochloride, the guanylyl cyclase inhibitor 1H-[1, 2, 4]oxadiazolo[4,3-a]quinoxalin-1-one, the cyclooxygenase inhibitor indomethacin, or K(+) channel blockers such as tetraethylammonium ions, glibenclamide, and BaCl2. Vindorosine-induced relaxations were attenuated in the presence of 0.1 µM nifedipine (an L-type Ca(2+) channel blocker). Vindorosine also concentration-dependently suppressed contractions induced by CaCl2 (0.01-5 mM) in Ca-free 60 mM KCl solution. Furthermore, fluorescence imaging using fluo-4 demonstrated that 30 min incubation with 100 µM vindorosine reduced the 60 mM KCl-stimulated Ca(2+) influx in the smooth muscles of rat renal arteries. The present study is probably the first report of blood vessel relaxation by vindorosine and the possible underlying mechanisms involving the inhibition of Ca(2+) entry via L-type Ca(2+) channels in vascular smooth muscles.