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In this study, a combination of metabolomics and network pharmacology was used to study the pharmacodynamic substances and mechanism of action of Yiyi Fuzi powder (YYFZ) on rheumatoid arthritis (RA) rats. The animal experiments were conducted in accordance with the requirements of the Experimental Animal Ethics Committee of Tianjin University of Traditional Chinese Medicine (approval number: TCM-LAEC2021241). The metabolomic analysis using UPLC-Q-TOF/MS technique identified 22 metabolites, including arachidonic acid, tryptophan, linoleic acid, phenylalanine, as significant biomarkers for the treatment of RA with YYFZ, and they were significantly regressed after YYFZ treatment. The analysis of YYFZ blood components also revealed that 11 blood components, including hypaconitine, benzoylhypaconitine, and deoxyaconitine, may be the components that exert direct pharmacological effects in YYFZ in vivo, and further network pharmacological analysis of blood components obtained that YYFZ may exhibit anti-inflammatory effects through acting on PI3K/Akt signaling pathway, estrogen signaling pathway, vascular endothelial growth factor (VEGF) signaling pathway. The results of this study provide implications for the clinical application of YYFZ.
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Fingerprints of lipophilic components in the roots of Salvia miltiorrhiza and S.yunnanensis were analyzed by UPLC-DADand UPLC coupled with mass spectroscopy to evaluate the differences and similarities of the lipophilic components in the two kinds of herbs.The UPLC analysis of 18 batches of S.miltiorrhiza and 16 batches of S.yunnanensis was performed on a 25℃Thermo Accucore C_(18)column(2.1 mm×100 mm,2.6μm)by Shimadzu LC-20AD;mobile phase was 0.026%phosphoric acid(A)-acetonitrile(B)with gradient elution;flow rate was 0.4 m L·min~(-1);detection wavelength was set at 270 nm;injection volume was 2μL.The molecular structures of the lipophilic components were analyzed on a 25℃Thermo Accucore C_(18)column(2.1 mm×100 mm,2.6μm)by Thermo U3000 UPLC Q Exactive Orbitrap LC-MS/MS with a mobile phaseconsisting of 0.1%formic acid water(A)and 0.1%formic acidacetonitrile(B).The mass spectrometry was acquired in positive modes using ESI.There are 10 common peaks in the lipophilic components of S.miltiorrhiza.The similarity between the 16 batches of S.miltiorrhiza and their own reference spectra was greater than 0.942,and the average similarity was 0.973.There are 12 common peaks in the lipophilic components of S.yunnanensis.The similarity between the 18 batches of S.yunnanensis and their own reference spectra was greater than 0.937,and the average similarity was 0.976.The similarity between the reference chromatograms of S.miltiorrhiza and S.yunnanensis was only 0.900.There are three lipophilic components in S.yunnanensis,which are not found in S.miltiorrhiza,and one of which isα-lapachone.There is a lipophilic component in S.miltiorrhiza not found in S.yunnanensis,which may be miltirone.The two herbs contain 8 common lipophilic components including dihydrotanshinoneⅠ,cryptotanshinone,tanshinoneⅠ,tanshinoneⅡ_A,nortanshinone in which the content of tanshinoneⅡ_A,dihydrotanshinoneⅠand cryptotanshinone of S.yunnanensisis significantly lower than that of S.miltiorrhiza(P<0.01),and the contents of tanshinoneⅠand nortanshinone are significantly lower than that of S.miltiorrhiza too(P<0.05).There are significant differences in the types and contents of lipophilic components between the roots of S.miltiorrhiza and S.yunnanensis,and the similarity between the fingerprints of interspecies is much lower than that between the same species.Therefore,the roots of S.miltiorrhiza and S.yunnanensis are two kinds of herbs which are quite different in chemical compounds and compositions.
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Chromatography, Liquid , Abietanes , Molecular Structure , Plant Roots , Salvia miltiorrhiza , Tandem Mass SpectrometryABSTRACT
Background@#The 47,XYY syndrome could result in fertility problems. However, seldom studies reported comprehensive researches on the embryonic development and pregnancy outcomes of these patients. This study aimed to evaluate the clinical outcomes of nonmosaic 47,XYY patients performed with fluorescent in situ hybridization (FISH) and preimplantation genetic diagnosis (PGD) treatment.@*Methods@#This was a retrospective study. Between January 2012 and May 2017, 51 infertile males with nonmosaic 47,XYY syndrome underwent FISH-PGD were included in the study. According to sex chromosomal FISH results, embryos were classified as normal signal, no nuclei fixed, no signal in fixed nuclei, suspensive signal, and abnormal signal groups, respectively. The incidence of each group, the fixation rate, and hybridization rate were calculated. Embryonic development and pregnancy outcomes were also analyzed. The measurement data were analyzed with Student's t-test. The comparison of categorical data was analyzed with the Chi-square test and Fisher's exact test when expected cell count was 0.05), and were significantly lower than the normal signal group (66.4%, P < 0.001). The clinical pregnancy rates of fresh and frozen embryos transferred cycles were 70.6% and 85.7%, respectively.@*Conclusions@#Among embryos with a clear diagnosis of sex chromosome, about one-fifth showed abnormal signals. Embryos with two sex chromosomal signals are more likely to develop into good-quality ones. The application of the PGD by FISH may help to improve the clinical outcome s.
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Female , Humans , Male , Pregnancy , In Situ Hybridization, Fluorescence , Infertility, Male , Genetics , Preimplantation Diagnosis , Retrospective Studies , Sex Chromosome Disorders , Diagnosis , Genetics , XYY Karyotype , Diagnosis , GeneticsABSTRACT
The fatty acid contents in soybean oil, corn oil, coconut oil, sunflower seed oil and fish oil were detected by gas chromatography. According to the characteristics of fatty acids of raw oil and the relevant regulations of GB 10765-2010,the formula of 6 kinds of human milk lipid substitutes was designed by Matlab software and Excel linear programming. At the same time, the Schaal oven accelerated oxidation method was used to study the oxidation process of 6 formulations using synchronous fluorescence technique combined with traditional chemical reagents (oxidation index:peroxide value,value of fennel and total oxidation value). By statistical analysis of data, the statistical relationship between fluorescence intensity and oxidation index was explored. The results showed that the main fatty acids of coconut oil were lauric acid and myristic acid,and its saturated fatty acid content was the highest 93.75% ±0.06%;while all the other oils were mainly composed of palmitic acid,linoleic acid and other components,wherein the fish oil had the highest content of unsaturated fatty acid. With the increase of oxidation time,the fluorescence intensity decreased gradually and the oxidation index value increased continuously. The correlation analysis and regression analysis results showed that the change of oxidation index and the fluorescence intensity of 6 samples had negative correlation, and the fluorescence intensity and the peroxide value and total oxidation value was quadratic linear correlation, and anisidine value as a linear correlation had strong correlation.
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Comparative analysis of the variations in HA 1 gene of the influenza A (H3N2) virus and the vaccine recommended were conducted in Shangluo city of China,during the surveillance year of 2014-2015.In this study,we collected the samples of H3N2 subtype strain from the Shanglou City of China during the surveillance period of 2014-2015.The strain was cultured in MDCK cells,HA gene fragment was amplified by RT-PCR and the nucleotide sequence was determined.Sequence alignment was performed using the clustax2.1 software.The phylogenetic tree was constructed by Mega6.0 software and was analyzed by Neighboring-joining method.Results showed that the homology of isolated strain during 2014-2015 was 97.2 ％-99.9％ and homology with the recommended vaccine strain A/Texas/50/2012 was 97.3％-98.5％.The amino acid sequence of the HA 1 gene of the isolated strain was compared with that of the vaccine strain.The major antigenic determinants of the isolates in 2014,having mutations were section B,Y159F,S198P,while the major antigenic determinants of isolates in 2015,having amino acid mutations were A zone G142R,B region S159F,S198P.These results indicated that the key antigenic determinant of influenza H3N2 subtype strain in Shangluo City has changed in 2014-2015 and A/Texas/50/2012 vaccine component is no more effective.Hence,there is an urgent need to update the influenza H3N2 subtype vaccine components and in future we should be deeply concerned about the evolution ofinfluenza H3N2 gene trends.
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Objective To analysis the effects on the growth of rats by repeated restraint in dermal exposure test. Methods SD rats in the restraint group was bound for 6 hours per day for 91 days according the way by dermal exposure, while SD rats in the control group didn't receive the treatment. Clinical signs, body weight and food consumption changes were observed for 91 days. When the study was terminated, hematology, clinical biochemistry, urinalyses, gross necropsy, and histopathology were carried out. Statistical methods such as the generalized estimating equation were used to compare the differences between two groups. Results The statistical results of generalized estimating equation showed that there was an interaction between the group and test time for male and female rats in body weight changes (P<0.05), and the body weight of male rats in the restraint group was lower than the control group (P<0.05) . Further analysis showed that for male rats there was significant difference between groups since the forth week (P<0.05), and the interaction was found between groups and test time (P<0.05) . For female rats, the interaction was found since the eighth week between the group and test time (P <0.05) .There was no significant differences in other parameters between two groups (P>0.05) . Conclusion Repeated restraint during dermal exposure affected the body weight gain of rats, and the sensitivity of male rats was higher than that of female rats.
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<p><b>OBJECTIVE</b>To study the effect of calcium-sensing receptor (CaSR) agonists and antagonists on the expression of CaSR in neonatal mice with persistent pulmonary hypertension (PPHN), and to clarify the role of CaSR in neonatal mice with PPHN.</p><p><b>METHODS</b>Forty-nine neonatal mice were randomly divided into four groups: control (n=10), hypoxia (PPHN; n=11), agonist (n=13), and antagonist (n=15). The mice in the PPHN, agonist, and antagonist groups were exposed to an oxygen concentration of 12%, and those in the control group were exposed to the air. The mice in the agonist and antagonist groups were intraperitoneally injected with gadolinium chloride (16 mg/kg) and NPS2390 (1 mg/kg) respectively once daily. Those in the PPHN and the control groups were given normal saline daily. All the mice were treated for 14 consecutive days. Hematoxylin and eosin staining and immunohistochemistry were used to observe the changes in pulmonary vessels. Laser confocal microscopy was used to observe the site of CaSR expression and measure its content in lung tissues. qRT-PCR and Western blot were used to measure the mRNA and protein expression of CaSR in lung tissues.</p><p><b>RESULTS</b>Compared with the control group, the PPHN group had significant increases in the pulmonary small artery wall thickness and the ratio of right to left ventricular wall thickness (P<0.05), which suggested that the model was successfully prepared. Compared with the control group, the PPHN group had a significant increase in the mRNA and protein expression of CaSR (P<0.05), and the agonist group had a significantly greater increase (P<0.05); the antagonist group had a significant reduction in the mRNA and protein expression of CaSR (P<0.05).</p><p><b>CONCLUSIONS</b>CaSR may play an important role in the development of PPHN induced by hypoxia in neonatal mice.</p>
Subject(s)
Animals , Mice , Hypoxia , Lung , Pathology , Myocardium , Pathology , Persistent Fetal Circulation Syndrome , Pathology , Pulmonary Artery , Pathology , RNA, Messenger , Receptors, Calcium-Sensing , Genetics , PhysiologyABSTRACT
Objective To understand the developmental effects induced by CdSe /ZnS quantum dots(QDs)on zebrafish embryos.Methods Zebrafish embryos were exposed to 0,0.5,1,2,4,8 and 16 nmol/L of CdSe /ZnS QDs,and the typical toxicological indexes were recorded at five time points respectively (24 hours post fertilization (hpf),48 hpf, 72 hpf,96 hpf,120 hpf).Results The results showed that the median lethal concentration (LC50 )for zebrafish embryos after 120 hpf was 21.38 nmol/L(95% CI =17.21 -26.57).The frequency of spontaneous movement in 60 seconds after 24 hpf,the frequency of heart beat in 60 seconds after 48 hpf,the hatching rate and the mortality rate were obviously affected by CdSe /ZnS QDs.Several abnormalities and toxic symptoms caused by CdSe /ZnS QDs at 8 nmol/L and 16 nmol/L were observed including pericardial edema,liver atrophy,non -depleted yolk,intestinal abnormal development and muscle degeneration after 120 hpf.Conclusion High level of CdSe /ZnS QDs (more than 8 nmol/L)could induce toxic effects on zebrafish embryonic development.
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Objective To evaluate acute toxicity and genotoxicity of sapindus saponin and to provide toxicological basis for sapindus saponin ’s daily applications. Methods Acute oral toxicity test,mammalian erythocyte micronucleus test, bacterial reverse mutation test and in vitro mammalian chromosome aberration test were used to investigate the effect of the sapindus saponin on gene mutation and chromosome aberration in both prokaryotic and eukaryotic cells. Results The acute toxicity test showed that the LD50 of sapindus saponin was 4640 mg/kg for both male and female mice. Toxic symptoms were observed including salivation,mucus and other toxic manifestations. There was no significant difference between the each dose group and the negative control group in the results of mammalian erythocyte micronucleus test( P>0. 05). The results of bacterial reverse mutation test were also negative. In each dose group and strain with or without S9,the number of revertant colonies did not exceed 2 times than that of spontaneous revertant colonies( negative control). No dose-response relationship was observed. The vitro mammalian chromosome aberration test showed that the IC50 of sapindus saponin on CHL was 75 μg/ml,and the differences between each dose group and the negative control group were not statistically significant( P >0. 05 ). However,the positive control group differed from the negative control group in all tests( P <0. 01). Conclusion Under this experimental condition,sapindus saponin has no genotoxicity.
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AIMS@#To investigate the antitumor effects of extracts from Oxytropis falcata on human hepatocellular carcinoma SMMC-7721 cells in vitro and in transplanted murine H22 tumors in vivo.@*METHODS@#Cell proliferation, cell cycle distribution and apoptosis in SMMC-7721 cells were determined and tumor growth inhibition in H22 tumors was investigated. Cell cycle distribution was analyzed by flow cytometry with propidium iodide (PI) and Annexin V-FITC/ PI double staining.@*RESULTS@#MTT assay revealed that essential oil and flavonoids of O. falcata (named EOOF and FOF) inhibited proliferation of SMMC-7721 cells in a dose-dependent manner. The IC50 value of EOOF and FOF were 0.115 and 0.097 mg·mL(-1), respectively. Cell cycle was arrested at G(1) phase, and induction of apoptosis occurred in SMMC-7721 cells when subjected to FOF. Growth inhibition in H22 solid tumors transplanted mice was significantly pronounced after being treated with FOF, and the inhibition ratio were 56.1% and 70.8% at the concentration of 30 and 60 mg·kg(-1).@*CONCLUSION@#The results suggest that FOF promotes apoptosis in SMMC-7721 cells and inhibits H22 tumor growth, resulting in a potential antitumor effect on hepatic cancer.
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Animals , Humans , Male , Mice , Antineoplastic Agents, Phytogenic , Apoptosis , Carcinoma, Hepatocellular , Drug Therapy , Cell Cycle , Cell Line, Tumor , Cell Proliferation , Drugs, Chinese Herbal , Growth Inhibitors , Liver Neoplasms , Drug Therapy , Mice, Inbred ICR , Oxytropis , ChemistryABSTRACT
Objective: The callus induction and subculture condition of Taxus chinensis var. mairei were optimized in the experiment. High Taxol-yielding callus was obtained, and the correlation between expression patterns of Taxol biosynthesis related genes and Taxol content was defined. Methods: Optimal hormone composition and concentration for induction and subculture of callus derived from T. chinensis var. mairei were briefly studied. The growth characteristics and Taxol content of calli derived from T. chinensis var. mairei (including vitro embryos, juvenile stems, and juvenile buds), T. media (vitro embryos) and T. brevifolia (vitro embryos) were compared. Expression of Taxol biosynthesis related genes in the above different calli was analyzed. Results MS Medium supplemented with 3.0 mg/L 2,4-D was suitable for callus induction of T. chinensis var. mairei, the induced rate is up to 92%. Subculture medium supplemented with 2.0 mg/L 2,4-D and 1.0 mg/L 6-BA was suitable for Taxol biosynthesis of T. chinensis var. mairei. Under the same culture conditions, Taxol content of calli derived from vitro embryos of T. chinensis var. mairei was the highest and had reached 0.027% of callus dry weight. The genes of Taxol biosynthesis related enzymes - geranylgeranyl diphosphate synthase (GGPPS), taxadiene synthase (TASY), taxane-10β-hydroxylase (T10βH), 10-deacetylbaccatin III-β-10-O-acetyltransferase (DBAT), phenylalanine aminomutase (PAM), and 3′-N-debenzoyltaxol N-benzoyltransferase (DBTNBT), were expressed at the high level in the high Taxol-producing callus. Conclusion: Vitro embryos can be used as the first choice expiant source to obtain high Taxol-yielding callus. To improve gene expression level of GG-PPS, TASY, T10βH, DBAT, PAM, and DBTNBT would promote Taxol biosynthesis.
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Luteolin is an important member of the flavonoid family. It has been reported that luteolin can inhibit the proliferation of serials of tumor cells including solid tumor, ascites cancer and human myeloid leukemia. Luteolin can also sensitize a number of apoptosis-inducing factors by unique mechanisms.
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Animals , Humans , Angiogenesis Inhibitors , Pharmacology , Antineoplastic Agents, Phytogenic , Pharmacology , Apoptosis , Luteolin , Pharmacology , Neoplasms , Pathology , Tumor Cells, CulturedABSTRACT
<p><b>OBJECTIVE</b>To observe the effects of kaempferol and quercetin on the activity of cytochrome P450 in rat hepatocytes.</p><p><b>METHODS</b>Primarily cultured rat hepatocytes were exposed to kaempferol or quercetin in concentrations of 0.1, 1, 10 micromol/L for 12 h, 24 h and 48 h. Hepatocytes CYP isoemzymes-erythromycin N-demethylase (ERND) and aminopyrine N-demethylase (ADM) activities were determined by Nash methods. Erythromycin (10 micromol/L) was used as positive control and DMSO(0.1%) as solvent control.</p><p><b>RESULTS</b>Kaempferol and quercetin inhibited ENRD activity in a dose-and time-dependent manner. In dose-response study, the ENRD activities in kaempferol (0.1,1 and 10 micromol/L) treated groups were (0.088+/-0.008), (0.074+/-0.006) and (0.041+/-0.003)micromol/(mg.min(-1)), respectively. ENRD activity in quercetin treated groups at the same concentrations were (0.082+/-0.007), (0.063+/-0.007) and (0.034+/-0.005) micromol/(mg.min(-1)), respectively. In time-courses study, the ENRD activity exposed to 10 micromol/L kaempferol or quercetin for 12 h and 48 h were (0.053+/-0.006) and (0.037+/-0.007) micromol/(mg.min(-1)), or (0.067+/-0.005) and (0.032+/-0.004) micromol/(mg.min(-1)). ADM activity was inhibited only by kaempferol in 10 mol/L at 24 h, but was not significantly altered by quercetin at any concentration tested.</p><p><b>CONCLUSION</b>In the present condition, kaempferol and quercetin act as potential CYP3A4 inhibitors as they can significantly inhibit ENRD in primarily cultured rat hepatocytes.</p>