Influence of traditional Chinese medicines on the in vivo metabolism of lopinavir/ritonavir based on UHPLC-MS/MS analysis.

A fast, reliable, and cost-effective liquid chromatography-tandem mass spectrometry method was established to determine the effects of the traditional Chinese medicine employed to treat coronavirus disease 2019, namely, Lianhua Qingwen granules, Huoxiang Zhengqi capsules, Jinhua Qinggan granules, Shufeng Jiedu capsules, and Angong Niuhuang pills, on the pharmacokinetics of lopinavir/ritonavir in rats. Blood samples were prepared using the protein precipitation method and atazanavir was selected as the internal standard (IS). Separation was performed on an Agilent ZORBAX eclipse plus C18 (2.1 mm × 50 mm, 1.8 µm) column using acetonitrile and water containing 0.1% formic acid as the mobile phase for gradient elution. The flow rate was 0.4 mL/min and the injection volume was 2 µL. Agilent Jet Stream electrospray ionization was used for mass spectrometry detection under positive ion multiple reaction monitoring mode at a transition of m/z 629.3→447.3 for lopinavir, m/z 721.3→296.1 for ritonavir, and m/z 705.4→168.1 for the IS. The method showed good linearity in the concentration range of 25-2500 ng/mL (r=0.9981) for lopinavir and 5-500 ng/mL (r=0.9984) for ritonavir. The intra-day and inter-day precision and accuracy were both within ±15%. Items, such as dilution reliability and residual effect, were also within the acceptable limits. The method was used to determine the effects of five types of traditional Chinese medicines on the pharmacokinetics of lopinavir/ritonavir in rats. The pharmacokinetic results showed that the half-life of ritonavir in the groups administered Lianhua Qingwen granules and Huoxiang Zhengqi capsules combined with lopinavir/ritonavir was prolonged by approximately 1.5- to 2-fold relative to that in the control group. Similarly, the pharmacokinetic parameters of lopinavir were altered. Overall, the results of this study offer important theoretical parameters for the effective clinical use of five types of traditional Chinese medicines combined with lopinavir/ritonavir to reduce the occurrence of clinical adverse reactions.

Network Pharmacology Analysis on the Mechanism of Huangqi Sijunzi Decoction in Treating Cancer-Related Fatigue.

OBJECTIVE: This study aimed to determine the active ingredients of Huangqi Sijunzi Decoction (HQSJZD) and the targets in treating cancer-related fatigue (CRF) so as to investigate the treatment mechanism of HQSJZD for CRF. METHODS: This study adopted the method of network pharmacology. The active ingredients and targets of HQSJZD were retrieved, and the targets of HQSJZD in treating CRF were obtained using a Venn diagram. Next, a protein-protein interaction (PPI) network was constructed using the String database. The core targets of HQSJZD in treating CRF were identified through topological analysis, and functional annotation analysis and pathway enrichment analysis were carried out. Subsequently, a compound-disease-target regulatory network was constructed using Cystoscape 3.8.0 software. RESULTS: A total of 250 targets of HQSJZD ingredients, 1447 CRF-related genes, and 144 common targets were obtained. Through topological analysis, 61 core targets were screened. Bioinformatics annotation of these genes identified 2366 gene ontology (GO) terms and 172 enriched Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. CONCLUSION: The active ingredients in HQSJZD, that is, quercetin, luteolin, kaempferol, and naringenin, may act on AKT1, IL-6, VEGFA, MAPK3, CASP3, JUN, and EGFR to regulate the PI3K-Akt, TNF, and IL-17 signaling pathways, thereby suppressing inflammatory response, tumor gene expression, and tumor angiogenesis to treat CRF. This study investigated the pharmacological basis and mechanism of HQSJZD in the treatment of CRF using systematic pharmacology, which provides an important reference for further elucidation of the anti-CRF mechanism and clinical applications of HQSJZD, and also provides a method protocol for similar studies in the future.

The synergistic antifungal effects of gypenosides combined with fluconazole against resistant Candida albicans via inhibiting the drug efflux and biofilm formation.

The increased resistance of Candida to conventional antifungals brings great challenges for the clinical treatment of Candida infections. Recently, more attention has been paid to the research on combination therapy, which is a potential therapeutic approach for overcoming Candida resistance. In the present study, we first investigated the interaction between gypenosides (Gyp) and fluconazole (FLC) against Candida albicans (C. albicans) in vitro and in vivo. The in vitro test revealed a synergistic antifungal activity between Gyp and FLC against FLC-resistant (FLCR) C. albicans and indifferent effects for FLC-susceptible (FLCS) C. albicans, with the fractional inhibitory concentration index of 0.2539-0.2578 and 1-1.5, respectively. Besides, Gyp displayed synergistic interaction with FLC against FLCRC. albicans performed biofilm over 4 h, with the fractional inhibitory concentration index <0.5. In vivo, the combined antifungal efficacy of Gyp with FLC was evaluated by Galleria mellonella (G. mellonella) larvae. Gyp plus FLC prolonged the survival rate and reduced tissue invasion of larvae infected with FLCRC. albicans. Further experiments to get a first hint at what antifungal mechanisms might be inhibition of early biofilm formation, suppression of drug efflux, and inhibition of yeast-hyphal conversion. These findings will provide a new approach for the treatment of C. albicans infection.

Efficacy of Eutectic Mixture of Local Anesthetics on Pain Control During Extracorporeal Shock Wave Lithotripsy: A Systematic Review and Meta-Analysis.

BACKGROUND The efficacy of a eutectic mixture of local anesthetics (EMLA) for pain control in extracorporeal shock wave lithotripsy is unclear. The aim of this study was to assess the effect of EMLA cream on pain control during extracorporeal shock wave lithotripsy. MATERIAL AND METHODS We searched Medline, EMBASE, and the Cochrane Central Register of Controlled Trials to identify relevant randomized controlled trials that compared the pain control efficacies of EMLA vs. placebo. Study eligibility criteria, participants, and interventions: Randomized controlled trials that compared the effect of EMLA with placebo cream for patients underwent extracorporeal shock wave lithotripsy. Study appraisal and synthesis methods: Two review authors extracted data independently using a designed data extraction form and risk of bias by Cochrane Collaboration's tool. RESULTS Nine studies, including 10 randomized controlled trials with 1167 patients, were eligible. The EMLA group experienced less pain (mean difference, -0.47; 95% confidence interval, -0.78 to -0.16; p=0.003) and shorter duration of lithotripsy (mean difference, -1.70, 95% confidence interval: -2.31 to -1.10, p<0.0001) than the placebo group. There were no significant differences in the number of patients who needed extra intravenous medication (p=0.610), number of patients with insufficient extracorporeal shock wave lithotripsy pain control (p=0.530), and number of patients with opioid adverse effects (p=0.320). Limitations: Long interval between the studies, different kinds of lithotripters. CONCLUSIONS EMLA can reduce pain during the ESWL procedure.

[Apoptosis and Ca~(2+) balance disorder of BV-2 cells induced by polychlorinated biphenyl].

OBJECTIVE: To study the neurotoxicity and mechanism of polychlorinated biphenyl( PCB) in BV-2 cells. METHODS: The experiment was divided into control group and PCB118, 138, 153, 180 treated group, the dosages were 0. 000, 0. 015, 0. 030, 0. 050, 0. 100, 0. 150 µmol/L. Through the statistical analysis of the survival rate of the cells, the final dose was divided into low, middle and high dose levels. Among them, the high dose group except PCB118 was 0. 15 µmol/L, the PCB138, 153, 180 concentration was 0. 015, 0. 05 and 0. 1 µmol/L for the low, medium and high dose groups. In vitro culture of mouse microglia cells with different doses of PCB after exposure, CCK-8 method to detect cell growth and viabilities; FITC Annexin V/PI method to detect cell apoptosis rate; detection of lactate dehydrogenase( LDH) release by ELISA kit; fluorescence probe method( Fluo-4 AM) to detect the intracellular calcium content changes. RESULTS: CCK-8 and FITC Annexin V/PI double staining experiments showed that PCB118, 138, 153, 180 could inhibit cell activity, and with the increase in dose, cell survival rate decreased, apoptosis rate increased. PCB118 dose in 0. 15 µmol/L toxicity increased, cytoactive was reduced to( 66. 56 ± 0. 10) %, apoptosis rate increased to( 27. 39 ± 1. 80) %; PCB138, 153 in 0. 1 µmol/L cell survival rate reached( 66. 66 ± 0. 10) % and( 67. 17 ± 0. 12) %, apoptosis rate reached( 48. 77 ± 2. 43) % and( 56. 42 ± 3. 59) %; PCB180 dose of 0. 015µmol/L cell survival rate reached( 92. 07 ± 0. 38) %, cell apoptosis rate was( 6. 82 ±0. 51) %, significantly higher than that of the control group, the difference was statistically significant( P < 0. 05). LDH assay showed that the release amount of LDH was proportional to the dose of PCB. The lactate dehydrogenase amounts of PCB118, 138, 153 and 180 were( 523. 78 ± 13. 58) U/L, ( 430. 37 ± 22. 03) U/L, ( 540. 58 ± 13. 08)U/L, ( 411. 88 ± 21. 92) U/L, which were significantly higher than those in the control group, the difference was statistically significant( P < 0. 05). The result of Fluo-4 and AM showed that the average fluorescence intensity of intracellular calcium( Ca~(2+)) in PCB exposed group increased obviously. The average fluorescence intensity of Ca~(2+) in the PCB118, 138, 153 and 180 groups was( 10. 14 ± 2. 36), ( 32. 47 ± 1. 56), ( 16. 54 ± 0. 97)and( 40. 46 ± 2. 19) when the dose was 0. 015 µmol/L, significantly higher than that of the control group, the difference was statistically significant( P < 0. 05). CONCLUSION: PCB could cause neurotoxicity damage, increase LDH, destroy cell membrane integrity, induce the balance disorder of cell calcium and lead to the cell apoptosis.

Clavuridins A and B, two new trinor-guaiane sesquiterpenes isolated from the Xisha soft coral Clavularia viridis.

In the present study, two new trinor-guaiane sesquiterpenes, named clavuridins B (1), and A (2), along with three known sesquiterpenes (3-5), were isolated from the Xisha soft coral Clavularia viridis. Their structures and absolute configurations were determined on the basis of spectroscopic analysis, X-ray diffraction analysis with Cu Kα radiation and by comparison with related model compounds. Compounds 1 and 3-5 were evaluated for their cytotoxic activity.

Protective Effects of Dietary Supplementation with a Combination of Nutrients in a Transgenic Mouse Model of Alzheimer's Disease.

OBJECTIVE: This study investigated the effects of intervention with a combination of nutrients in the amyloid precursor protein-presenilin (APP-PSN) C57BL/6J double transgenic mouse model of Alzheimer's disease (AD). METHODS: A total of 72 2-month-old APP-PSN mice were randomly assigned to three groups. The model group (MG) was fed regular, unsupplemented chow, while the low- and high-dose treatment groups (LG and HG, respectively) were given a combination of nutrients that included phosphatidylserine, blueberry extracts, docosahexaenoic acid, and eicosapentaenoic acid as part of their diet. An additional 24 wild-type littermates that were fed unsupplemented chow served as the negative control group (NG). After 3 and 7 months of treatment, the cognitive performance was assessed with the Morris water maze and the shuttle box escape/avoidance task, and the biochemical parameters and oxidative stress were evaluated in both the blood and brain. RESULTS: An improvement in antioxidant capacity was observed in the treatment groups relative to the MG at 3 months, while superior behavioral test results were observed in the mice of the HG and NG groups. In the MG, pycnosis was detected in neuronal nuclei, and a loss of neurons was observed in the cerebral cortex and the hippocampus. At 7 months, the ß-amyloid1-42 peptide accumulation was significantly elevated in the MG but was markedly lower in the mice fed the nutrient combination. The antioxidant capacity and behavioral test scores were also higher in these mice. CONCLUSIONS: Early intervention with a combination of nutrients should be considered as a strategy for preventing cognitive decline and other symptoms associated with AD.

Discovery and SAR studies of methionine-proline anilides as dengue virus NS2B-NS3 protease inhibitors.

A series of methionine-proline dipeptide derivatives and their analogues were designed, synthesized and assayed against the serotype 2 dengue virus NS2B-NS3 protease, and methionine-proline anilides 1 and 2 were found to be the most active DENV 2 NS2B-NS3 competitive inhibitors with Ki values of 4.9 and 10.5 µM. The structure and activity relationship and the molecular docking revealed that L-proline, L-methionine and p-nitroaniline in 1 and 2 are the important characters in blocking the active site of NS2B-NS3 protease. Our current results suggest that the title dipeptidic scaffold represents a promising structural core to discover a new class of active NS2B-NS3 competitive inhibitors.

Beneficial effects of Yerba Mate tea (Ilex paraguariensis) on hyperlipidemia in high-fat-fed hamsters.

Yerba Mate tea (Mate), an infusion made from the leaves of the tree Ilex paraguariensis, is a widely consumed beverage in South America. Mate has previously been shown to have hypolipidemic effects. However, its mechanism of action is not well understood. This study was conducted to determine the effect of Mate on hyperlipidemia induced in hamsters by a high-fat diet, as well as its mechanism of action. Fifty male hamsters were randomly assigned to normal control, high-fat control, and high-fat with Mate tea aqueous extract (1%, 2% or 4% w/v) groups. We evaluated the effects of Mate aqueous extract on body weight, serum lipids, antioxidant enzyme activity, lipoprotein metabolism enzyme activity, and gene expression involved in lipid metabolism in hyperlipidemic hamsters. Mate aqueous extract significantly decreased body-weight gain and lowered serum lipid levels in the hyperlipidemic hamster model. Meanwhile, Mate treatment increased antioxidant enzyme activity, improved lipoprotein lipase (LPL) and hepatic lipase (HL) activities in serum and liver, upregulated mRNA expression of peroxisome proliferator-activated receptor α and low density lipoprotein receptor, and downregulated mRNA expression of sterol regulatory element-binding protein 1c and acetyl CoA carboxylase in the liver. The results indicate that Mate tea ameliorates hyperlipidemia partly by reducing lipid peroxidation, improving endothelial function and LPL and HL activities, and modulating the expression levels of genes involved in lipid oxidation and lipogenesis.

The determination of persistent organic pollutants (POPs) in the colostrums of women in preterm labor.

BACKGROUND: Human milk not only contains nutrients and antibodies, but also can be used as an indicator for levels of organic pollutants in human bodies. We developed a method for determining persistent organic pollutants (POPs) in the colostrum of women in preterm labor, the POPs of 36 colostrum samples have been examined. METHODS: Thirty-six samples of colostrum from preterm women were extracted by acetone-acetonitrile, enriched and purified by solid-phase Florisil columns. The purified POPs were further separated by the capillary columns, and detected by the gas chromatography-electron capture detection (GC-ECD). RESULTS: The average recovery rates of 6 types of organochlorine-based pesticides were 80.2%-112.1%, which represent the first 3 categories of the 12 species of POPs. The precise quantities detected were 3.85%-9.32% (the limits of detection were 0.03 microg/l to 0.08 microg/l), and the linear correlation coefficients were >or=0.9969. Of the 36 women tested, 10 (27.8%) were found to have colostrums containing traces of dichloro-diphenyl-trichloroethane (DDT), and 2 (5.56%) were tested positive for dieldrin. CONCLUSIONS: The combination of using GC-ECD proved to be both accurate and reliable, and this process proved to be both simple and time-effective. This method is applicable for determining the levels of POPs in organisms.