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BACKGROUND: Fusarium graminearum is a devastating fungal pathogen that poses a significant threat to global wheat production and quality. Control of this toxin-producing pathogen remains a major challenge. This study aimed to isolate strains with antagonistic activity against F. graminearum and at the same time to analyze the synthesis of deoxynivalenol (DON), in order to provide a new basis for the biological control of FHB. RESULTS: Total of 69 microorganisms were isolated from the soil of a wheat-corn crop rotation field, and an antagonistic bacterial strain F12 was identified as Burkholderia pyrrocinia by molecular biology and carbon source utilization. F. graminearum control by strain F12 showed excellent biological activities under laboratory conditions (95.8%) and field testing (63.09%). Meanwhile, the DON content of field-treated wheat grains was detected the results showed that F12 have significantly inhibited of DON, which was further verified by qPCR that F12 produces secondary metabolites that inhibit the expression of DON and pigment-related genes. In addition, the sterile fermentation broth of F12 not only inhibited mycelial growth and spore germination, but also prevented mycelia from producing spores. CONCLUSION: In this study B. pyrrocinia was reported to have good control of FHB and inhibition of DON synthesis. This novel B. pyrrocinia F12 is a promising biological inoculant, providing possibilities for controlling FHB, and a theoretical basis for the development of potential biocontrol agents and biofertilizers for agricultural use. © 2024 Society of Chemical Industry.
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The potential anti-stroke active components in Taohong Siwu Decoction(THSWD) were identified by target cell trapping coupled with ultra-high performance liquid chromatography-quadrupole-time of flight mass spectrometry(UPLC-Q-TOF-MS). The underlying mechanism of active components in THSWD in the treatment of ischemic stroke(IS) was explored by network pharmacology, molecular docking, and experimental validation. The UPLC-Q-TOF-MS technology combined with the UNIFI data analysis platform was used to analyze the composition of the cellular fragmentation fluid after co-incubation of THSWD with target cells. The targets of potential active components and IS were collected by network pharmacology, and the common targets underwent protein-protein interaction(PPI), Gene Ontology(GO), and Kyoto Encyclopedia of Genes and Genomes(KEGG) signaling pathway enrichment analyses. The target cell trapping component-core target-signaling pathway network was constructed, and the active components were molecularly docked to the top targets in the PPI network, followed by pharmacodynamic validation in vitro. Fifteen active components were identified in the target cellular fragmentation fluid, including bicyclic monoterpenes, cyanoglycosides, flavonols, quinoid chalcones, phenylpropanoids, and tannins. As revealed by the analysis of network pharmacology, THSWD presumably regulated PI3K-AKT, FoxO, MAPK, Jak-STAT, VEGF, HIF-1, and other signaling pathways to affect inflammatory cascade reaction, angiogenesis, oxidative stress, pyroptosis, apoptosis, and other pathological processes via paeoniflorin, butylphthalide, dehydrated safflower yellow B, 3,4-dicaffeoylquinic acid, amygdalin, paeoniflorin, and ligusticolactone. Molecular docking and in vitro pharmacodynamic validation revealed that the target cell trapping active components could promote neovascularization in rat brain microvascular endothelial cells(rBMECs) in the oxygen-glucose deprivation/reoxygenation(OGD/R) model. The application of target cell trapping coupled with UPLC-Q-TOF-MS technology can rapidly screen out the potential active components in THSWD. The active components of THSWD can be predicted to intervene in the pathogenesis of IS through network pharmacology, and molecular docking combined with experimental validation can further clarify the efficacy, thus providing a theoretical basis for research ideas on the pharmacodynamic substance basis of traditional Chinese medicine compounds.
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Animales , Ratas , Accidente Cerebrovascular Isquémico/tratamiento farmacológico , Simulación del Acoplamiento Molecular , Farmacología en Red , Células Endoteliales , Fosfatidilinositol 3-Quinasas , Medicamentos Herbarios Chinos/farmacologíaRESUMEN
Objective: To investigate the type, length, and CG loci of HBV DNA CpG islands in HBsAg positive maternal C genotype and its relationship with intrauterine HBV transmission, so as to provide a new perspective for the study of intrauterine transmission of HBV. Methods: From June 2011 to July 2013, HBsAg-positive mothers and their newborns who delivered in the obstetrics and gynecology department of the Third People's Hospital of Taiyuan were collected. Epidemiological data were collected through face-to-face questionnaires and electronic medical records. Serum HBV markers and serum HBV DNA were detected by electrochemiluminescence and quantitative fluorescence PCR, respectively. Intrauterine transmission of HBV was determined by positive HBsAg and/or HBV DNA in femoral venous blood before injection of HBV vaccine/Hepatitis B immunoglobulin within 24 h of birth. A total of 22 mothers and their newborns with HBV DNA load ≥106 IU/ml in intrauterine transmission were selected as the intrauterine transmission group, and 22 mothers with HBV DNA load ≥106 IU/ml without intrauterine transmission were chosen as the control group by random seed method. The distribution prediction of CpG islands of HBV DNA in 39 mothers with genotype C by HBV DNA sequencing was analyzed. Results: Among 39 mothers with HBV C genotype, 19 were in the intrauterine transmission group, and 20 were in the control group. The HBV DNA of 39 patients with genotype C traditional CpG island Ⅱ and Ⅲ, while the control group had traditional CpG island Ⅰ and novel CpG island Ⅳ and Ⅴ. The length of CpG island Ⅱ and Ⅲ and the number of CG loci of CpG island Ⅱ in the intrauterine transmission group differed from those in the control group (P<0.05). The CpG island Ⅱ length ≥518 bp and the number of CG loci ≥40 in the intrauterine transmission group (11/19) were significantly higher than those in the control group (2/20) (P<0.05). The length of CpG island Ⅱ and the number of CG loci in the X gene promoter region (Xp region) were higher than those in the control group (P<0.05). In the HBV intrauterine transmission group, most of maternal (12/19) HBV DNA CpG island Ⅱ completely covered the Xp region, which was significantly higher than that in the control group (5/20), and the number of HBV DNA Xp region CG loci was higher than that in the control group (P<0.05). Conclusions: The distribution of maternal C genotype HBV DNA CpG islands is related to intrauterine transmission. The length of CpG island Ⅱ and the number of CG sites may increase the risk of intrauterine transmission of HBV.
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Femenino , Humanos , Recién Nacido , Embarazo , Biomarcadores , Islas de CpG , ADN Viral/genética , Hepatitis B , Antígenos de Superficie de la Hepatitis B , Virus de la Hepatitis B/genética , Transmisión Vertical de Enfermedad Infecciosa , Madres , Complicaciones Infecciosas del EmbarazoRESUMEN
The present study investigated the therapeutic effect and mechanism of Di'ao Xinxuekang(DXXK) on non-alcoholic steatohepatitis(NASH) in mice. Sixty-five C57 BL/6 J mice were randomly divided into a normal group and an experimental group for model induction with the high-fat diet for 16 weeks. Then the mice in the experimental group were randomly divided into a model group, an atorvastatin group(4 mg·kg~(-1)·d~(-1)), and high-(200 mg·kg~(-1)·d~(-1)), medium-(60 mg·kg~(-1)·d~(-1)), and low-dose(20 mg·kg~(-1)·d~(-1)) DXXK groups, with 10 mice in each group. Drugs were administered by gavage for eight weeks. Serum lipid, liver lipid, serum alanine aminotransferase(ALT), aspartate aminotransferase(AST), malondialdehyde(MDA), superoxide dismutase(SOD), and glutathione reductase(GSH-Px) were determined. Interleukin-1β(IL-1β) and tumor necrosis factor-α(TNF-α) were measured by enzyme-linked immunosorbent assay(ELISA). The liver index was calculated. The liver pathological change and lipid accumulation were observed by HE and oil red O staining. The liver ultrastructure was observed by the transmission electron microscope. The mRNA and protein expression of nuclear factor-erythroid 2 related factor 2(Nrf2) and heme oxygenase-1(HO-1) was detected by real-time fluorescence-based quantitative PCR and Western blot, respectively. The results showed that compared with the normal group, the model group displayed serum lipid and liver lipid metabolism disorders, elevated transaminase, lipid deposition, steatosis, and inflammation, suggesting that the NASH model in mice was properly induced. Compared with the model group, the DXXK groups showed decreased serum lipid, liver lipid, ALT, AST, MDA, IL-1β, and TNF-α, increased SOD and GSH-Px, alleviated hepatic steatosis, ballooning, and inflammation, and up-regulated Nrf2 and HO-1 gene and protein expression. In conclusion, DXXK can significantly alleviate NASH in mice, which is related to the inhibition of oxidative stress and inflammatory damage by up-regulating the Nrf2/HO-1 signaling pathway.
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Animales , Ratones , Medicamentos Herbarios Chinos , Inflamación/metabolismo , Lípidos , Hígado , Factor 2 Relacionado con NF-E2/metabolismo , Enfermedad del Hígado Graso no Alcohólico/metabolismo , Estrés Oxidativo , Transducción de Señal , Superóxido Dismutasa/metabolismo , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
OBJECTIVES@#To investigate the effects of circ_0005379 on the proliferation, apoptosis, migration, and invasion of oral squamous cell carcinoma (OSCC) cells and its mechanism.@*METHODS@#Real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect the expression levels of circ_0005379 and miR-17-5p in OSCC tissues and SCC15 cell lines. Western blot was used to detect the expression levels of acyl-CoA oxidase 1 (ACOX1). The circ_0005379 overexpression vector was transfected into SCC15 cells. Methyl thiazolyl tetrazolium blue staining, flow cytometry, Transwell, and Western blot were used to detect the effects of circ_0005379 overexpression on the proliferation, apoptosis, migration, and invasion of SCC15 cells and the expression of E-cadherin, β-catenin, and Snail proteins. Dual luciferase reporter assay and RNA immunoprecipitation were used to examine the regulation of circ_0005379, miR-17-5p, miR-17-5p, and ACOX1 in SCC15 cells. A nude mouse xenograft model of SCC15 cells stably overexpressing circ_0005379 was established, and the effect of circ_0005379 overexpression on the growth of xenografts in nude mice was observed.@*RESULTS@#Compared with adjacent cancer tissues, the expression levels of circ_0005379 and ACOX1 proteins in OSCC tissues were decreased (@*CONCLUSIONS@#circ_0005379 may inhibit the proliferation, migration, and invasion of OSCC cells by downregulating the expression of miR-17-5p and upregulating ACOX1, which promote apoptosis and inhibit tumor growth
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Animales , Humanos , Ratones , Acil-CoA Oxidasa , Carcinoma de Células Escamosas/genética , Proliferación Celular , Neoplasias de Cabeza y Cuello , Ratones Desnudos , MicroARNs , Neoplasias de la Boca/genética , ARN Circular , Carcinoma de Células Escamosas de Cabeza y CuelloRESUMEN
In this study, a high performance thin-layer chromatography/single quadrupole mass spectrometry QDa (HPTLC-QDa) method for robust authentication of Ganoderma lucidum, a popular and valuable herbal medicine, has been developed. This method is simple and practical, which allows direct generation of characteristic mass spectra from the HPTLC plates automatically with the application of in situ solvent desorption interface. The HPTLC silica gel plates were developed with toluene-ethyl formate-formic acid (5 : 5 : 0.2, V/V) and all bands were transferred to QDa system directly in situ using 80% methanol with 0.1% formic acid as desorption solvent. The acquired HPTLC-QDa spectra showed that luminous yellow band b3, containing ganoderic acid B/G/H and ganodeneric acid B, the major active components of Ganoderma, could be found only in G. lucidum and G. lucidum (Antler-shaped), but not in G. sinense and G. applanatum. Moreover, bands b13 and b14 with m/z 475/477 and m/z 475/491/495, respectively, could be detected in G. lucidum (Antler-shaped), but not in G. lucidum, thus allowing simple and robust authentication of G. lucidum with confused species. This method is proved to be simple, practical and reproducible, which can be extended to analyze other herbal medicines.
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A comprehensive analytical method based on ultra-fast liquid chromatography coupled with triple quadrupole/linear ion trap tandem mass spectrometry(UFLC-QTRAP-MS/MS) was established for simultaneous determination of the content of 38 active components in Abelmoschi Corolla, including flavonoids, organic acids, nucleosides and amino acids, so as to investigate the effects of different harvesting and processing methods on multi-active components in Abelmoschi Corolla. The chromatographic separation was performed on a XBridg®C_(18) column(4.6 mm×100 mm, 3.5 μm) with(0.1% formic acid water) methanol-acetonitrile(1∶1) as the mobile phase for gradient elution at 30 ℃. The flow rate was 0.5 mL·min~(-1). The components were detected in a multiple-reaction monitoring(MRM) mode. The gray relational analysis(GRA) was used to comprehensively evaluate the multiple active components of Abelmoschi Corolla at different harvesting times and drying temperatures. The results showed that 38 components had a good linearity with correlation coefficients all above 0.999 0. The method featured a good precision, repeatability and stability with the relative stan-dard deviations(RSDs) of less than 5.0%. Recoveries ranged from 98.06% to 104.4% with RSD between 0.22% and 4.9%. The results of GRA indicated that a better quality in the samples collected on September 9 th. Samples dried at 90 ℃ had a better quality. The established method is accurate and reliable, and can be used to assess the internal quality of Abelmoschi Corolla. This study can provide basic materials for determining appropriate harvesting time and processing method of Abelmoschi Corolla.
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Aminoácidos , Cromatografía Líquida de Alta Presión , Cromatografía Liquida , Nucleósidos , Espectrometría de Masas en TándemRESUMEN
Nonalcoholic fatty liver disease (NAFLD) is a metabolic stress-induced liver injury characterized by excessive lipid accumulation in hepatocytes, which is closely related to insulin resistance and genetic susceptibility. It falls into the category of "liver lump" in traditional Chinese medicine (TCM). NAFLD affects about 25% of the population worldwide and has become a major burden of the world health care system. However, its exact pathogenesis remains unclear. Conducting the basic research on NAFLD is of great clinical significance and social value. As an important tool for NAFLD research, animal model plays a particularly important role in clarifying the pathophysiological mechanism of NAFLD. In recent years, the modeling methods for NAFLD in China and abroad have been constantly updated, and in particular, certain progress has been made in the duplication of TCM syndrome models. By consulting and sorting out the relevant literature published in recent years in China and abroad, the author summarized the replication methods of NAFLD animal models. This paper reviewed the advantages and disadvantages of models established via dietary induction (high-fat feed, high-fat and high-fructose feed, high-fat and high-cholesterol feed, and methionine choline-deficient feed), models with genetic defects [leptin-deficiency (Lepob/Lepob), autosomal recessive diabetes gene homozygous deficiency (ob/ob), Alms1 gene (foz/foz) mutation, and FATZO mice] and exposure to special diets, and models for TCM syndromes (liver depression and spleen deficiency syndrome, phlegm-dampness syndrome, blood stasis syndrome, combined phlegm and stasis syndrome, and qi stagnation and blood stasis syndrome), in order to provide reference for the preparation of more scientific, reasonable, economical, and convenient animal models of NAFLD, thus laying a foundation for in-depth study of the pathogenesis, prevention, and treatment of NAFLD.
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Objective:To study the effect of Wenjing Huayu Zhitong therapy (Xiangyan Zhitong prescription, XZP) on the mitogen activated protein kinase (MAPK)/extracelluar regulated protein kinase (ERK) signaling pathway of primary dysmenorrhea (PD) rats, and explore the pathogenesis of PD and the mechanism of action of Wenjing Huayu Zhitong therapy. Method:Forty-eight female SPF-grade Wistar rats were randomly divided into blank group,model group, western medicine group, low-dose XZP group, medium-dose XZP group, and high-dose XZP group, with 8 rats in each group. In addition to the blank group, dysmenorrhea rat model with cold coagulation and blood stasis syndrome was established by cold stimulation combined with estradiol benzoate and oxytocin. The rats in the blank group,model group,western medicine group, low-dose XZP group, medium-dose XZP group, and high-dose XZP group were given distilled water, distilled water,0.06 g·kg<sup>-1</sup> ibuprofen, 6.55 g·kg<sup>-1</sup> XZP, 13.09 g·kg<sup>-1</sup> XZP, and 26.18 g·kg<sup>-1</sup> XZP, respectively, by gavage for 6 days. The writhing latency and writhing frequency of rats were recorded within 30 min after oxytocin injection.Western blot was used to detect the protein expression of B-Raf, mitogen activates extracellular regulated kinases1/2 (MEK1/2), extracellular regulated kinases1/2 (ERK1/2), p-MEK1/2, p-ERK1/2, c-Jun, and cyclooxygenase-2 (COX-2) in rat uterus. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was employed to detect the mRNA expression of B-Raf, MEK1, MEK2, ERK1, ERK2, c-Jun, and cyclooxygenase-2 (COX-2) in rat uterus. Result:Compared with the model group,the treatment groups showed insignificantly prolonged writhing latency and significantly reduced writhing frequency (<italic>P</italic><0.01). On the 6<sup>th</sup> day of modeling, there was no significant difference in the quantitative scores of symptoms and signs among the treatment groups. On the 12<sup>th</sup> day of modeling, the scores changed little in the western medicine group and the low-dose XZP group and decreased significantly in the medium- and high-dose XZP groups (<italic>P</italic><0.01) compared with those in the model group. Compared with those in the blank group, the protein and mRNA levels of p-MEK1/2, p-ERK, B-Raf, c-Jun, and COX-2 in the model group were significantly up-regulated (<italic>P</italic><0.01). Compared with those in the model group, the protein and mRNA levels of p-MEK1/2, p-ERK1/2, B-Raf, c-Jun, and COX-2 in the western medicine group, medium-dose XZP group, and high-dose XZP group were significantly down-regulated (<italic>P</italic><0.05,<italic>P</italic><0.01). Conclusion:The mechanism of Wenjing Huayu Zhitong therapy in treating PD with cold coagulation and blood stasis syndrome may be related to the down-regulation of MAPK/ERK signaling pathway.
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Objective:To investigate the effects of Di'ao Xinxuekang (DXXK) on NLRP3 inflammasome in mouse RAW264.7 macrophages and thoracic aorta of rats with atherosclerosis (AS), so as to explore its anti-AS mechanism. Method:RAW264.7 cells were stimulated with oxidized low density lipoprotein (ox-LDL) and then intervened with MCC950 and DXXK. The contents of tumor necrosis factor-<italic>α</italic> (TNF-<italic>α</italic>) and interleukin-1<italic>β</italic> (IL-1<italic>β</italic>) were determined by enzyme linked immunosorbent assay (ELISA). The mRNA and protein expression levels of Nod-like receptor protein 3 (NLRP3), inflammasome adaptor protein apoptosis-associated speck-like protein containing CARD (ASC), and cysteine-dependent aspartate-directed protease-1 (Caspase-1) were detected by real-time polymerase chain reaction (Real-time PCR) and Western blotting. Sixty male SD rats were randomly divided into the normal group, model group, atorvastatin group (2.0 mg·kg<sup>-1</sup>), as well as high-, medium-, and low-dose (100, 30, and 10 mg·kg<sup>-1</sup>) DXKK groups, with 10 rats in each group. The rats were exposed to the high-fat diet and vitamin D<sub>2</sub> for inducing AS. The blood lipid level was measured using an automatic biochemical analyzer, followed by the calculation of AS index (AI). The contents of serum TNF-<italic>α</italic> and IL-1<italic>β</italic> were determined by ELISA, and the mRNA and protein expression levels of NLRP3, ASC, and Caspase-1 in thoracic aorta were assayed by Real-time PCR and Western blotting. HE staining and Sirius red staining were conducted to observe the pathomorphological changes in the abdominal aorta and aortic sinus. Result:Compared with the normal group, the model group exhibited significantly increased TNF-<italic>α</italic> and IL-1<italic>β</italic> contents and up-regulated NLRP3, ASC, and Caspase-1 mRNA and protein expression in RAW264.7 cells (<italic>P</italic><0.01). The above indexes in each drug administration group were significantly reduced in contrast to those in the model group (<italic>P</italic><0.05, <italic>P</italic><0.01). The comparison with the model group showed that cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), and AI in each DXXK group significantly declined, while the high-density lipoprotein cholesterol (HDL-C) was significantly elevated (<italic>P</italic><0.05, <italic>P</italic><0.01). The levels of serum TNF-<italic>α</italic> and IL-1<italic>β</italic> and the mRNA and protein expression levels of NLRP3, ASC, and Caspase-1 in the thoracic aorta were decreased (<italic>P</italic><0.05, <italic>P</italic><0.01). Abdominal aortic lesions and fibrous hyperplasia of aortic sinus were significantly improved. Conclusion:DXXK has a significant anti-AS effect, which is possibly related to the inhibition of NLRP3 inflammasome.
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Aim To investigate the mechanism of therapeutic effect of ginsenoside metabolite compound K (GCK) on imiquimod-induced psoriasis mice. Methods Model mice induced by imiquimod (IMQ) were randomly divided into the following groups; model group, GCK (14, 28, 56, 112 mg • kg-
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Aim To investigate the effect of astragalin (AG) on airway inflammation in asthmatic mice and its mechanism. Methods Fifty SPF male mice were randomly divided into normal group, asthma model group, and astragalin low (AG25), medium (AG50), and high (AG100) dose groups. A mouse model of asthma was prepared by egg albumin inhalation, the number of cells was counted by Diff-Quik staining after collecting bronchoalveolar lavage fluid (BALF), and IL4, 5, and 13 levels in BALF were measured by ELISA. The inflammatory changes in mouse lung tissues were observed using HE staining. Reactive oxygen species (ROS) levels were measured using a DCFH-DA probe, and superoxide dismutase (SOD) activity and malondialdehyde (MDA) content were measured by colorimetry. IL-4, IL-5, IL-13, NOX2, p47phox, p-NF-κBp65, NF-κBp65, IκBα, p-IκBα and β-actin expressions in lung tissues were detected by Western blot. Results AG significantly reduced the number of inflammatory cells and total cells in BALF, decreased IL-4, IL-5, and IL-13 contents in BALF and lung tissues, and reduced inflammatory cell infiltration in lung tissues. AG inhibited nicotinamide adenine dinucleotide phosphate oxidase 2 (NOX2) and p47phox expression, decreased ROS levels and MDA levels, increased SOD activity, and inhibited IκBα and NF-κBp65 phosphorylation. Conclusion AG attenuates airway inflammation in asthma by modulating the oxidative stress response through the NOX2/ROS/NF-κB signaling pathway.
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In this study, the chemical profiling of Jingyin Granules and the tissue distribution of nine major constituents in this Chinese medicine were performed after oral administration of Jingyin Granules to rats, by using UHPLC-Q-Exactive Orbitrap HR-MS. An Acquity UPLC BEH C_(18) chromatographic column(2.1 mm×100 mm, 1.7 µm) was used as solid phase, while the mobile phase was methanol and 0.1% formic acid water for gradient elution. The major constituents in this Chinese medicine were quickly and accurately identified, via comparison with the retention times and MS/MS spectra of the standards. A total of 106 chemicals were identified from Jingyin Granules, including 24 kinds of organic acids, 47 kinds of flavonoids, 10 kinds of iridoids, and 21 kinds of saponins and 4 kinds of other compounds. After oral administered Jingyin Granules to rats, 48, 30, 25, 23, 45, 34, 39, 26, 19 prototype compounds were identified in serum, heart, liver, spleen, lung, kidney, brain, fat, and testicles, respectively. Meanwhile, an LC-MS based analytical method was established for simultaneous determination of chlorogenic acid, swertiamarin, caffeic acid, sweroside, liquiritin, prim-O-glucosylcimifugin, arctiin, 5-O-methylvisammioside and arctigenin in biological samples. The tissue distribution(serum, liver and lung) of these nine aim constituents in rats after oral administration of Jingyin Granules were investigated. It was found that these nine constituents could be quickly absorbed into circulation system and then distributed to liver and lung tissues. Except arctigenin, the exposure of other eight aim constituents to serum and lung was peaked at 1 h. At 1 h, the exposure of these components to lung tissue were ranked as follows: swertiamarin [(75 191.0±3 483.21) ng·g~(-1)]>arctiin [(2 716.5±36.06) ng·g~(-1)]>5-O-methylvisammioside [(585.1±0.71) ng·g~(-1)]>arctigenin [(437.45±3.18) ng·g~(-1)]>chlorogenic acid [(308.1±5.66) ng·g~(-1)]>prim-O-glucosylcimifugin [(211.35±2.19) ng·g~(-1)]>sweroside [(184.3±9.05) ng·g~(-1)]>caffeic acid [(175.95±2.05) ng·g~(-1)]>liquiritin [(174.78±153.34) ng·g~(-1)]. In summary, an UHPLC-Q-Exactive Orbitrap HR-MS method has been established for rapid and accurate identification of the constituents in Jingyin Granules, while the tissue distribution of nine major absorpted constituents were investigated in rats following oral administration of Jingyin Granules. These findings provided key information and guidance for further studies on pharmacodynamic substances and clinical applications of Jingyin Granules.
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Medicamentos Herbarios Chinos , Espectrometría de Masas en Tándem , Animales , Cromatografía Líquida de Alta Presión , Cromatografía Liquida , Ratas , Distribución TisularRESUMEN
The human UDP-glucuronosyltransferase 1A1 (UGT1A1), one of the most essential conjugative enzymes, is responsible for the metabolism and detoxification of bilirubin and other endogenous substances, as well as many different xenobiotic compounds. Deciphering UGT1A1 relevance to human diseases and characterizing the effects of small molecules on the activities of UGT1A1 requires reliable tools for probing the function of this key enzyme in complex biological matrices. Herein, an easy-to-use assay for highly-selective and sensitive monitoring of UGT1A1 activities in various biological matrices, using liquid chromatography with fluorescence detection (LC-FD), has been developed and validated. The newly developed LC-FD based assay has been confirmed in terms of sensitivity, specificity, precision, quanti-tative linear range and stability. One of its main advantages is lowering the limits of detection and quantification by about 100-fold in comparison to the previous assay that used the same probe substrate, enabling reliable quantification of lower amounts of active enzyme than any other method. The precision test demonstrated that both intra- and inter-day variations for this assay were less than 5.5%. Further-more, the newly developed assay has also been successfully used to screen and characterize the regu-latory effects of small molecules on the expression level of UGT1A1 in living cells. Overall, an easy-to-use LC-FD based assay has been developed for ultra-sensitive UGT1A1 activities measurements in various biological systems, providing an inexpensive and practical approach for exploring the role of UGT1A1 in human diseases, interactions with xenobiotics, and characterization modulatory effects of small mole-cules on this conjugative enzyme.
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Public health crises, such as the outbreak of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) since Dec. 2019, are widely acknowledged as severe traumatic events that impose threats not only because of physical concerns but also because of the psychological distress of infected patients. We designed an internet-based integrated intervention and evaluated its efficacy on depression and anxiety symptoms in patients infected by SARS-CoV-2.
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Adulto , Femenino , Humanos , Masculino , Persona de Mediana Edad , Ansiedad , Terapéutica , Betacoronavirus , Teléfono Celular , China , Infecciones por Coronavirus , Psicología , Depresión , Terapéutica , Internet , Atención Plena , Pandemias , Neumonía Viral , Psicología , Estudios Prospectivos , Distrés Psicológico , Terapia por Relajación , Autocuidado , MétodosRESUMEN
OBJECTIVE@#To explore the biomechanical characteristics of "three-dimensional balanced manipulation" for the treatment of cervical spondylotic radiculopathy(CSR).@*METHODS@#A CSR patient was treated with "three-dimensional balanced manipulation", and the mechanical changes during the manipulation were monitored by mechanical testing system. Using spiral CT to scan the neck of the patient to obtain DICOM data. The three-dimensional finite element model of cervical spondylotic radiculopathy was established by using Mimics software, Geographic Studio software. The "three-dimensional balance manipulation" was simulated and loaded, and the mechanical parameters of each part were replaced into the finite element model, and the finite element analysis was carried out by using ANSYS software to study the internal stress changes and displacement deformation of vertebral body and intervertebral disc under the action of "three-dimensional balance manipulation".@*RESULTS@#The established C-C finite element model of the CSR patient consisted of 5 vertebrae, 4 intervertebral discs and 3 ligaments, involving 153 471 nodes and 64 978 units. The stress of C-C vertebral body was mainly located in anterior and root of C spinous processes, arch, vertebral arch and the combination of the two after full loading of manipulation, and the maximum stress was 17.781 MPa. The deformation sites were mainly concentrated in articular processes and anterior transverse processes of C, superior articular processes and transverse processes of C, articular processes of C. The stress of C-C intervertebral disc mainly distributed in the anterior part of C intervertebral disc and the nucleus pulposus of C and C. The displace mentextended to the middle and posterior part of C nucleus pulposus, around the nucleus of C and C and anterior part of cervical intervertebral disc.@*CONCLUSION@#The establishment of three-dimensional finite element model of C-C cervical spondylotic radiculopathy can simulate the geometry and material properties of cervical spine, and also accurately reflects the biomechanical characteristics of cervical spine, verifys the internal mechanism of "three-dimensional balanced manipulation" on CSR, proves the safety and effectiveness of treatment, guides more standardized manipulation, and avoids medical accidents.
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Humanos , Fenómenos Biomecánicos , Vértebras Cervicales , Análisis de Elementos Finitos , Disco Intervertebral , Radiculopatía , Rango del Movimiento Articular , EspondilosisRESUMEN
In this study, the chemical profiling of Jingyin Granules and the tissue distribution of nine major constituents in this Chinese medicine were performed after oral administration of Jingyin Granules to rats, by using UHPLC-Q-Exactive Orbitrap HR-MS. An Acquity UPLC BEH C_(18) chromatographic column(2.1 mm×100 mm, 1.7 μm) was used as solid phase, while the mobile phase was methanol and 0.1% formic acid water for gradient elution. The major constituents in this Chinese medicine were quickly and accurately identified, via comparison with the retention times and MS/MS spectra of the standards. A total of 106 chemicals were identified from Jingyin Granules, including 24 kinds of organic acids, 47 kinds of flavonoids, 10 kinds of iridoids, and 21 kinds of saponins and 4 kinds of other compounds. After oral administered Jingyin Granules to rats, 48, 30, 25, 23, 45, 34, 39, 26, 19 prototype compounds were identified in serum, heart, liver, spleen, lung, kidney, brain, fat, and testicles, respectively. Meanwhile, an LC-MS based analytical method was established for simultaneous determination of chlorogenic acid, swertiamarin, caffeic acid, sweroside, liquiritin, prim-O-glucosylcimifugin, arctiin, 5-O-methylvisammioside and arctigenin in biological samples. The tissue distribution(serum, liver and lung) of these nine aim constituents in rats after oral administration of Jingyin Granules were investigated. It was found that these nine constituents could be quickly absorbed into circulation system and then distributed to liver and lung tissues. Except arctigenin, the exposure of other eight aim constituents to serum and lung was peaked at 1 h. At 1 h, the exposure of these components to lung tissue were ranked as follows: swertiamarin [(75 191.0±3 483.21) ng·g~(-1)]>arctiin [(2 716.5±36.06) ng·g~(-1)]>5-O-methylvisammioside [(585.1±0.71) ng·g~(-1)]>arctigenin [(437.45±3.18) ng·g~(-1)]>chlorogenic acid [(308.1±5.66) ng·g~(-1)]>prim-O-glucosylcimifugin [(211.35±2.19) ng·g~(-1)]>sweroside [(184.3±9.05) ng·g~(-1)]>caffeic acid [(175.95±2.05) ng·g~(-1)]>liquiritin [(174.78±153.34) ng·g~(-1)]. In summary, an UHPLC-Q-Exactive Orbitrap HR-MS method has been established for rapid and accurate identification of the constituents in Jingyin Granules, while the tissue distribution of nine major absorpted constituents were investigated in rats following oral administration of Jingyin Granules. These findings provided key information and guidance for further studies on pharmacodynamic substances and clinical applications of Jingyin Granules.
Asunto(s)
Animales , Ratas , Cromatografía Líquida de Alta Presión , Cromatografía Liquida , Medicamentos Herbarios Chinos , Espectrometría de Masas en Tándem , Distribución TisularRESUMEN
Ganjiang Lingzhu Decoction is one of the first 100 classical prescriptions published by China in 2018. According to the published literature, it was found that there is no review on the history, evolution and research progress of this prescription. In order to reflect the history, modifications, quality control and clinical applications, this paper focuses on the origination, evolution, current development and modern studies of Ganjiang Lingzhu Decoction, in the hope of providing a reference for the heritage and innovation of other classical prescriptions.
Asunto(s)
China , Medicamentos Herbarios Chinos , Medicina Tradicional China , Prescripciones , Control de CalidadRESUMEN
Objective:To observe the clinical efficacy of Wenjing Huayu Zhitong therapy in treating primary dysmenorrhea (PD) with cold coagulation and blood stasis, and to explore its immune mechanisms on PD. Method:The 108 PD patients with cold coagulation and blood stasis syndrome were collected and randomly divided into traditional Chinese medicine (TCM) group, ibuprofen group and placebo group according to the random number table method, with 36 cases in each group. All patients received corresponding medicines three days before menstruation. The patients in TCM group were treated with TCM and ibuprofen sustained release capsule simulator. The patients in ibuprofen group were treated with ibuprofen sustained-release capsule and TCM simulator. The patients in placebo group were treated with TCM simulator and ibuprofen sustained-release capsule simulator. Treatment lasted for 6 consecutive days for three menstrual cycles, and follow-up was conducted for three menstrual cycles after drug withdrawal. The visual analogue score (VAS), total time of abdominal pain and TCM symptom scores in each menstrual cycle were recorded. The levels of CD3+, CD4+, CD8+ and the ratio of CD4+/CD8+ in peripheral blood before and after treatment were detected by flow cytometry. Result:After treatment for three menstrual cycles, both the TCM group and ibuprofen group were better than placebo group in reducing VAS score and reducing total abdominal pain time (P<0.01). The long-term follow-up effect after drug withdrawal in TCM group was significantly better than that in ibuprofen group (P<0.01). The total effective rate was 91.43% (32/35) in TCM group, 66.67% (10/33) in ibuprofen groups, and 30.30% (10/33) in placebo group . The efficacy of the TCM group was better than that of the ibuprofen group (χ2=-2.971, P<0.01), and the efficacy of the ibuprofen group was better than that of the placebo group (χ2=-2.371, P<0.05). After treatment, the levels of CD3+, CD4+ and CD4+/CD8+ in TCM group were significantly increased and the levels of CD8+ were decreased significantly as compared with those before treatment (P<0.01). After treatment, the levels of CD4+ and CD4+/CD8+ in TCM group were higher (P<0.05,P<0.01),while the levels of CD8+ were significantly lower than those in ibuprofen group and placebo group (P<0.01). Conclusion:Wenjing Huayu Zhitong therapy can reduce the VAS score and accumulative time of abdominal pain, and improve the dysmenorrhea symptoms in patients with PD. Reversal of the T cell subsets disorder may be one of its mechanisms in treating PD with cold coagulation and blood stasis.
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Objective: To establish the quality control methods for the standard decoction of Puerariae Thomsonii Radix.Method:According to the preparation principles for traditional Chinese medicine (TCM) standard decoction,13 batches of Puerariae Thomsonii Radix from different origins were analyzed under the chromatography conditions established in this study and verified with methodology.By referring to Chinese Pharmacopoeia of 2015,puerarin was used as a quantitative indicator to calculate the transfer rate.In this study,the structures of main chromatographic peaks were also identified to clarify the main chemical constituents in the standard decoction.Result:The 13 batches of medicinal herbs were identified as Puerariae Thomsonii Radix,with a recovery rate of 98.0%,and RSD of 1.1%,indicating that the method was accurate and reliable.The transfer rate ranged from 41.4% to 60.0%,and the extraction rate was within the range of 15.7%-34.3%.The corresponding fingerprints were prepared for 13 batches of the standard decoction,and their similarities were all greater than 90.0%.The chemical constituents from Puerariae Thomsonii Radix were identified by mass spectrometry analysis,including citric acid,4'-O-glucosyl puerarin/daidzein-4',7-diglucoside,3'-hydroxy puerarin/genistein puerarin,puerarin apioside,daidzin puerossid A and daidzein,etc.Conclusion: The 13 batches of Puerariae Thomsonii Radix decoction in different origins had consistent properties with the basic properties of medicinal decoction pieces.The established method of quality evaluation can be used to systematically evaluate the standard decoction,providing reference for quality control of related decoction preparations of Puerariae Thomsonii Radix.