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1.
Article in Chinese | WPRIM | ID: wpr-928190

ABSTRACT

This study aims to explore the toxicity mechanism of Rhododendri Mollis Flos(RMF) based on serum metabolomics and network toxicology. The toxic effect of RMF on normal rats was evaluated according to the symptoms, serum biochemical indexes, and histopathology. Serum metabolomics was combined with multivariate statistical analysis to search endogenous differential metabolites and related metabolic pathways. The toxic components, targets, and signaling pathways of RMF were screened by network toxicology technique, and the component-target-metabolite-metabolic pathway network was established with the help of serum metabolomics. The result suggested the neurotoxicity, hepatotoxicity, and cardiotoxicity of RMF. A total of 31 differential metabolites and 10 main metabolic pathways were identified by serum metabolomics, and 11 toxic components, 332 related target genes and 141 main signaling pathways were screened out by network toxicology. Further analysis yielded 7 key toxic components: grayanotoxin Ⅲ,grayanotoxinⅠ, rhodojaponin Ⅱ, rhodojaponin Ⅴ, rhodojaponin Ⅵ, rhodojaponin Ⅶ, and kalmanol, which acted on the following 12 key targets: androgen receptor(AR), albumin(ALB), estrogen receptor β(ESR2), sex-hormone binding globulin(SHBG), type 11 hydroxysteroid(17-beta) dehydrogenase(HSD17 B11), estrogen receptor α(ESR1), retinoic X receptor-gamma(RXRG), lactate dehydrogenase type C(LDHC), Aldo-keto reductase(AKR) 1 C family member 3(AKR1 C3), ATP binding cassette subfamily B member 1(ABCB1), UDP-glucuronosyltransferase 2 B7(UGT2 B7), and glutamate-ammonia ligase(GLUL). These targets interfered with the metabolism of gamma-aminobutyric acid, estriol, testosterone, retinoic acid, 2-oxobutyric acid, and affected 4 key metabolic pathways of alanine, aspartate and glutamate metabolism, cysteine and methionine metabolism, steroid hormone biosynthesis, and retinol metabolism. RMF exerts toxic effect on multiple systems through multiple components, targets, and pathways. Through the analysis of key toxic components, target genes, metabolites, and metabolic pathways, this study unveiled the mechanism of potential neurotoxicity, cardiotoxicity, and hepatotoxicity of RMF, which is expected to provide a clue for the basic research on toxic Chinese medicinals.


Subject(s)
Animals , Cardiotoxicity , Chemical and Drug Induced Liver Injury , Drugs, Chinese Herbal/toxicity , Hormones , Metabolomics , Rats
2.
Article in Chinese | WPRIM | ID: wpr-906490

ABSTRACT

Objective:The volatile components of Rhododendri Mollis Flos were determined and the differences of volatile components at different flowering stages were compared and analyzed. Method:Gas chromatography-ion mobility spectrometry (GC-IMS) was used to detect the volatile components in Rhododendri Mollis Flos at different flowering stages (bud stage, initial flowering stage, half-flowering stage, blooming stage and late blooming stage). GC-IMS spectra combined with cluster analysis, principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) were used to compare the differences and similarities of volatile components in different flowering stages. Result:A total of 70 volatile components in Rhododendri Mollis Flos at different flowering stages were detected, among which 67 were common components, and 47 were identified qualitatively, mainly alcohols, esters and aldehydes. Carveol was a special component at the late blooming stage. The content of alpha-terpineol is the highest at the initial flowering stage, but not at the blooming stage and late blooming stage. The relative contents of the active ingredients [6-methyl-5-hepten-2-one, nonanal, alpha-terpineol, 1,8-cineole, linalool oxide, 1-octen-3-ol, (<italic>E</italic>)-3-hexenol] showed a decreasing trend during flowering stages. GC-IMS spectra showed that the samples at different flowering stages had their own characteristic peak regions, and also had common regions. The results of cluster analysis, PCA and OPLS-DA all showed that the samples at different flowering stages were distinguishable. OPLS-DA was used to screen 19 different components to distinguish different flowering stages, including <italic>γ</italic>-butyrolactone, 1,8-cineole, ethyl hexanoate, etc. Conclusion:Rhododendri Mollis Flos samples at different flowering stages can be distinguished obviously, and the active substances in the volatile components are gradually dissipated with the degree of flower opening, which can provide reference for the improvement of material basis and the study of different flowering stages of Rhododendri Mollis Flos.

3.
Article in Chinese | WPRIM | ID: wpr-906402

ABSTRACT

Objective:To analyze and identify the flavonoids of Citri Reticulatae Pericarpium with different aging time by an ultra-performance liquid chromatography-quadrupole-electrostatic field orbitrap high resolution mass spectrometry (UPLC-Q-Orbitrap HRMS). Method:Compounds were separated on Agilent Extend-C<sub>18</sub> column (3.0 mm×100 mm, 1.8 μm), mobile phase was 0.1% acetic acid aqueous solution (A)-0.1% acetic acid methanol solution (B) for gradient elution (0-25 min, 5%-95%B; 25-30 min, 95%B; 30-30.1 min, 95%-5%B; 30.1-35 min, 5%B), the flow rate was 0.4 mL·min<sup>-1</sup>, and the column temperature was set at 30 ℃. High resolution mass spectrometry was performed with electrospray ionization (ESI), and scanned in positive and negative ion modes by means of full scan/data dependent secondary scan (Full MS/dd-MS<sup>2</sup>). The multistage ion fragment information combined with mzCloud network database, local high resolution mass spectrometry database of traditional Chinese medicine components (OTCML), literature information and relevant reference materials were used for accurate qualitative analysis. Result:Totally 43 flavonoids in Citri Reticulatae Pericarpium were identified, including 24 flavones, 5 flavonols, 13 dihydroflavones and 1 chalcone. The flavonoids in samples with different aging time were basically consistent in material types, but the peak area was different. According to the comparison of relative content in the peak area, it was found that the relative contents of 30 flavonoids showed an overall increasing trend with the increase of aging time. Among them, the relative contents of 24 flavonoids (such as hesperidin, diosmin, 6-demethoxytangeretin, nobiletin and tangeretin) increased significantly. There was no significant change in the relative contents of the other 13 flavonoids (such as naringenin and neohesperidin). Conclusion:An efficient method is established in this paper to identify flavonoids in Citri Reticulatae Pericarpium with different aging time and their relative content changes rapidly and accurately. The findings provide a methodological reference for the study on pharmacodynamic material base and quality control of Citri Reticulatae Pericarpium, and it provides experimental basis that drugs processed long time ago have better effect of Citri Reticulatae Pericarpium.

4.
Article in Chinese | WPRIM | ID: wpr-905942

ABSTRACT

In this paper, through the collection and collation of ancient materia medica, medical books and medical formulary, combining with modern literature, the historical changes of the name, origin, position, harvesting time, medicinal parts, toxicity, functions and indications, processing methods of Rhododendri Mollis Flos (RMF) were systematically combed and verified, so as to provide reference for clinical application, processing standard and basic research of RMF. According to textual research, RMF is the dried flower of Rhododendron molle. In each historical period, there are many aliases and local names, being with phenomenon of homonyms and synonyms. RMF is mostly wild and planted in a small amount, harvesting time is mostly in March to April. However, the harvesting flowering period is differently described as initial bloom, full bloom and extensive bloom. RMF was first recorded in Shennong Bencaojing (《神农本草经》), but it did not mention its medicinal parts. Then the flowers, fruits, roots are be used as medicine, but flowers are still the main medicinal parts. RMF had a long processing history, included fried, vinegar-fried, wine-fried, steamed, wine-steamed, vinegar-steamed, and many other processing methods in ancient times. However, at present, only raw products are used in clinical practice, and only a few modern books retain the methods of stir-fried and wine-steamed, believing that the processing can reduce toxicity of RMF.

5.
Acta Pharmaceutica Sinica ; (12): 1041-1047, 2017.
Article in Chinese | WPRIM | ID: wpr-779692

ABSTRACT

This study was designed to investigate the correlation between idiosyncratic liver injury and content of cis-2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside(cis-SG)in radix Polygoni multiflori Preparata(RPMP). In order to compare the effect of hepatotoxicity of different cis-SG contents in RPMP, rats were administered with 50% alcohol extracts of RPMP(7.56 g·kg-1, via intragastric administration)alone or co-treated with lipopolysaccharide(LPS, 2.8 mg·kg-1, via tail vein injection). The results showed that no significant alterations of plasma ALT and AST activities were observed in the normal rats. In the LPS treated rats, the group without light treatment and the group with 0.10% cis-SG after light treatment did not exhibit obvious injury in liver. The group with 0.35% cis-SG after light treatment and the group with 0.70% cis-SG after light treatment showed significant increases in ALT, AST, TNF-α, IL-6, NF-κB p65 and apoptosis rate(P < 0.05), causing pathological changes in the liver tissue. Through the content analysis of drug in patients with liver injury, we found that the content of cis-SG(> 0.40%)was generally higher than that of pieces collected from different origins(< 0.10%). The comparative analysis of experiments and clinical data showed that there was a relationship between the content of cis-SG and idiosyncratic liver injury. In order to reduce the risk of clinical medication, the content of cis-SG of 0.10% should be a limit of quality control in the production processing of Polygonum multiflorum.

6.
Article in Chinese | WPRIM | ID: wpr-230063

ABSTRACT

Using six kinds of ionic liquids as extractants, ultrasonic-assisted extraction coupled with HPLC method was developed for the simultaneous determination of wilforgine, wiforizine, triptophenolide, wilforine and triptoquinone A in Tripterygium hypoglaucum. The separation was performed on an Inertsil ODS-4 column with the mobile phase of acetonitrile-0.1% phosphoric acid in gradient elution at a flow rate of 0.75 mL•min⁻¹. Detection wavelength was 220 nm and the column temperature was 30℃. Under the optimal extractions, the results showed that triptophenolide and triptoquinone A had the highest extraction yield by using 0.6 mol•L⁻¹ [BMIm]PF6 methanol solution as extraction solvent with the solid-liquid ratio of 1∶10. The calibration curves of triptophenolide and triptoquinone A showed a good linearity in the range of 0.000 65-0.026, 0.066 55-2.662 μg (r=0.999 9)respectively. The average recovery was 102.4% and 97.90% with RSD of 2.5% and 1.5%, respectively. Wilforgine, wiforizine and wilforine had the highest extraction yield when using 0.6 mol• L⁻¹ [BMIm]PF6absolute ethanol solution as extraction solvent with the solid-liquid ratio of 1∶10. The content of wilforgine, wiforizine and wilforine from 0.023 9-0.956, 0.002 7-0.108, 0.006 4-0.256 μg showed a good linearity (r=0.999 9), and the average recovery was 100.6%,99.50% and 98.70% with RSD of 2.1%,1.9% and 2.7%, respectively. The results indicated that this method is convenient, reliable and green, and can be used as a reliableanalytical method for the quality control of T.hypoglaucum.

7.
Article in Chinese | WPRIM | ID: wpr-275191

ABSTRACT

To investigate the dynamic change rules of volatile components from Atractylodis Macrocephalae Rhizoma with different stir-baking degrees (from slight stir-baking, stir-baking to yellow, stir-baking to brown, to stir-baking to scorch). In the present experiment, the Atractylodis Macrocephalae Rhizoma samples with different stir-baking degrees were collected at different processing time points. The contents of volatile oil in various samples were determined by steam distillation method, and the volatile compounds were extracted by using static headspace sampling method. Gas chromatography-mass spectrography (GC-MS) and automated mass spectral deconrolution and identification system (AMDIS) were combined with Kováts retention index to analyze the chemical constituents of the volatile compounds. The results showed that with the deepening of the stir-baking degree, the content of volatile oil was decreased step by step in 4 phases, and both the compositions and contents of volatile components from Atractylodis Macrocephalae Rhizoma showed significant changes. The results showed that the dynamic change rules of volatile components from Atractylodis Macrocephalae Rhizoma in the process of stir-baking were closely related to the processing degree; in addition, Atractylodis Macrocephalae Rhizoma and honey bran had adsorption on each other. These results can provide a scientific basis for elucidating the stir-baking (with bran) mechanism of Atractylodis Macrocephalae Rhizoma.

8.
Article in Chinese | WPRIM | ID: wpr-346171

ABSTRACT

<p><b>OBJECTIVE</b>To explore the abnormal expression of plasma proteins by analysis of proteomic expression profile in children with infectious mononucleosis (IM).</p><p><b>METHODS</b>Two dimensional gel electrophoresis (2-DE) followed by the mass spectrometry was used to examine important protein spots with different expression levels between children with IM and normal controls.</p><p><b>RESULTS</b>Seven differential proteins were obtained: hemopexin, vitamin D binding protein, fetuin A, C-reactive protein, apolipoprotein A, haptoglobin and transthyretin. Compared with the control group, haptoglobin showed a higher expression level in children with IM, and the expression levels of the other proteins were obviously down-regulated.</p><p><b>CONCLUSIONS</b>The expression changes of differential proteins identified in this study are all related with the liver acute injury, suggesting that children with IM are associated with acute liver injury. Further studies on the characteristics of above proteins will contribute to the diagnosis and treatment of pediatric IM.</p>


Subject(s)
Blood Proteins , Child , Child, Preschool , Electrophoresis, Gel, Two-Dimensional , Female , Humans , Infectious Mononucleosis , Blood , Male , Proteomics , Methods
9.
Article in Chinese | WPRIM | ID: wpr-236026

ABSTRACT

This study is to explore the reason of "the older, the better" of PCR and itsincrease of flavonoids. We identified the fun- gus isolated from the PCR using microscopic and molecular identification. HPLC method was used to determine the content of 4 fla- vonoids and to clarifythe regularity of them; UV spectrophotometry method was used to determine the total content of flavonoids; reverse thinking was applied to screen the fungus that have close relation to the change of flavonoids. Finally, we have isolated and identified 25 fungusfrom the PCR, which belong to 2 genus and 4 species, including pencillium commune, P. minioluteeum, P. citrinum, Aspergillus flavus and A. niger. The content of flavonoids was increased in the mildew PCR due to A. niger and other fungus. Therefore, "the ol- der, the better" of PCR had its scientific reason that the increase of flavonoids had a close relation of the metabolic activity of A. niger and other fungus.


Subject(s)
Citrus , Chemistry , Microbiology , Flavonoids , Fungi
10.
Article in Chinese | WPRIM | ID: wpr-246099

ABSTRACT

<p><b>OBJECTIVE</b>The multiple levels fragmentations of five furocoumarine (psoralen, xanthotoxin, bergapten, oxypeucedanin, and byakangelicol) in Angelica dahurica have been demonstrated using LTQ-Orbitrap mass spectrometry with high resolution and high mass accuracy to discover the possible,fragmentation regularity.</p><p><b>METHOD</b>Duringcollsion-induced dissociation (CID), the MS(n) data of the five compoundswhich were gained in the positive ion mode at 35ev collision energy by direct injection syrings method were analyzed using Xcalibar 2.0 Software to infer the formula of these fragmentations.</p><p><b>RESULT</b>The results indicated that the five compounds have similar fragmentation process with CO meutral lost at C5,C8-subsituents and furan ring, meanwhile the meutralloss of CO2 occurred easily at lactone group.</p><p><b>CONCLUSION</b>This method is helpful in identifying the structures of other furocoumarinein Angelica dahuricaand their metabolites in vivo.</p>


Subject(s)
Angelica , Chemistry , Chemical Phenomena , Coumarins , Chemistry , Drugs, Chinese Herbal , Chemistry , Mass Spectrometry , Methods , Molecular Structure
11.
Article in Chinese | WPRIM | ID: wpr-318690

ABSTRACT

<p><b>OBJECTIVE</b>To establish a derivative fluorometry method for the determination of sulfur dioxide residues in traditional Chinese medicine.</p><p><b>METHOD</b>The optimal derivation condition was established. The fluorescence intensity was detected at excitation wavelength of 321 nm, and emission wavelength of 384 nm.</p><p><b>RESULT</b>A linear relationship was obtained between the fluorescence intensity and the addition of reference substance in the range of 0.999 7-17.99 nmol with a correlation coeffient of 0.999 9, and the average recovery was 102.3% with RSD 4.6%.</p><p><b>CONCLUSION</b>This method is simple and sensitive with quick and correct result. It can provide a reference for the determination of sulfur dioxide residues in traditional Chinese medicine.</p>


Subject(s)
Drugs, Chinese Herbal , Fluorescence , Medicine, Chinese Traditional , Sensitivity and Specificity , Species Specificity , Spectrometry, Fluorescence , Methods , Sulfur Dioxide , Temperature
12.
Chinese Journal of Hepatology ; (12): 290-294, 2013.
Article in Chinese | WPRIM | ID: wpr-246692

ABSTRACT

<p><b>OBJECTIVE</b>To investigate whether the phosphorylation (functionally inhibitive) of eukaryotic initiation factor 2-alpha (eIF2-a) affects the molecular mechanism of cisplatin-induced cellular apoptosis in human hepatocellular carcinoma (HCC).</p><p><b>METHODS</b>The human HCC cultured cell lines SMMC-7221 and HepG2 were treated with cisplatin alone (controls; 24 h) or in combination with pre-transfection of a dominant-negative eIF2-a mutant (eIF2aS51A) or pre-exposure to an eIF2-a-specific phosphatase inhibitor (salubrinal) to decrease or increase the phosphorylation level, respectively. Changes in expression of apoptosis markers were quantitatively and qualitatively assessed by flow cytometry and western blot analysis. The significance of differences among groups was assessed by analysis of variance testing and of differences between groups was assessed by t-test.</p><p><b>RESULTS</b>Cisplatin treatment induced the appropriate functional-inhibitive phosphorylation of eIF2-a on serine 51. Cisplatin treatment (10 mg/ml) induced significant apoptosis in the eIF2aS51A pre-transfected SMMC-7721 (control: 21.7 +/- 1.5% vs. 50.7 +/- 2.1%, t = 19.454, P less than 0.05) and HepG2 (21.0 +/- 1.0% vs. 57.3 +/- 2.1%, t = 27.250, P less than 0.05). Salubrinal pre-treatment significantly inhibited the cisplatin (15 mg/ml)-induced apoptosis in SMMC-7721 (control: 50.3 +/- 2.5% vs. 16.3 +/- 2.1%, t = 18.031, P less than 0.05) and HepG2 (42.0 +/- 2.6% vs. 12.0 +/- 2.0%, t = 15.667, P less than 0.05).</p><p><b>CONCLUSION</b>Phosphorylation of eIF2-a may act to inhibit cisplatin-induced apoptosis of HCC.</p>


Subject(s)
Apoptosis , Carcinoma, Hepatocellular , Cell Line, Tumor , Cisplatin , Pharmacology , Eukaryotic Initiation Factor-2 , Metabolism , Humans , Liver Neoplasms , Phosphorylation
13.
Article in Chinese | WPRIM | ID: wpr-294067

ABSTRACT

This experiment shows how to use the automated mass spectral deconvolution & identification system (AMDIS) to deconvolve the overlapped peaks in the total ion chromatogram (TIC) of volatile oil from Chineses materia medica (CMM). The essential oil was obtained by steam distillation. Its TIC was gotten by GC-MS, and the superimposed peaks in TIC were deconvolved by AMDIS. First, AMDIS can detect the number of components in TIC through the run function. Then, by analyzing the extracted spectrum of corresponding scan point of detected component and the original spectrum of this scan point, and their counterparts' spectra in the referred MS Library, researchers can ascertain the component's structure accurately or deny some compounds, which don't exist in nature. Furthermore, through examining the changeability of characteristic fragment ion peaks of identified compounds, the previous outcome can be affirmed again. The result demonstrated that AMDIS could efficiently deconvolve the overlapped peaks in TIC by taking out the spectrum of matching scan point of discerned component, which led to exact identification of the component's structure.


Subject(s)
Chromatography , Methods , Citrus , Chemistry , Drugs, Chinese Herbal , Chemistry , Fruit , Chemistry , Mass Spectrometry , Methods , Oils, Volatile , Chemistry , Plant Oils , Chemistry
14.
Chinese Journal of Hepatology ; (12): 191-195, 2011.
Article in Chinese | WPRIM | ID: wpr-290605

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the role of miR-221/222 in inhibiting endoplasmic reticulum stress-induced human hepatocarcinoma cells apoptosis.</p><p><b>METHOD</b>miR-221/222 mimics and inhibitors were used to mimic or block the function of endogenous miR-221/222 respectively. Western blot and flow cytometry were used to test the effects of miR-221/222 on cell cycle and apoptosis under endoplasmic reticulum stress in human hepatocellular carcinoma cells.</p><p><b>RESULTS</b>Endoplasmic reticulum stress resulted in miR-221/222 down-regulation in human hepatocellular carcinoma cells. miR-221/222 mimics and inhibitors inhibited and promoted respectively endoplasmic reticulum stress-mediated p27Kip1 induction. Moreover, p27Kip1 suppression not only resulted in reduction in the fraction of G1 phase cells, but also promoted the endoplasmic reticulum stress-mediated apoptosis in human hepatocellular carcinoma cells.</p><p><b>CONCLUSION</b>miR-221/222 were downregulated by endoplasmic reticulum stress in human hepatocellular carcinoma cells, which subsequently protected human hepatocellular carcinoma cells against endoplasmic reticulum stress-induced apoptosis through p27Kip1 regulation.</p>


Subject(s)
Apoptosis , Carcinoma, Hepatocellular , Metabolism , Pathology , Cell Cycle , Cell Line, Tumor , Cyclin-Dependent Kinase Inhibitor p27 , Metabolism , Endoplasmic Reticulum , Metabolism , Humans , Liver Neoplasms , Metabolism , Pathology , MicroRNAs , Metabolism
15.
Article in Chinese | WPRIM | ID: wpr-299008

ABSTRACT

<p><b>OBJECTIVE</b>To study the essence of chronic viral hepatitis B (CHB) of damp-heat retention in the middle-jiao syndrome (DRMS) from plasma proteomic angle.</p><p><b>METHODS</b>Plasma proteomic analyses of plasma whole protein of patients in the group with CHB of DRMS (20 cases) and subjects in the health control group (5 cases) were compared using two-dimensional gel electrophoresis (2-DE), mass spectrography, and other bioinformatics analyses methods.</p><p><b>RESULTS</b>Eight protein dots with obvious regularity changes of differential expression were obtained by 2-DE. Seven protein dots were obtained by mass spectrography (One protein dot with undetected results): apolipoprotein C2 (APO-C2), vitronectin (VN), haptoglobin (HPT), transthyretin (TTHY), APO-A1, serum amyloid P-component (SAMP), and APO-A4. Compared with the health control group, the expressions of APO-A1 and APO-A4 were somewhat higher and the expressions of the expressions of the rest five protein dots were obviously down-regulated.</p><p><b>CONCLUSION</b>APO-Al and APO-A4 were of potential significance in the diagnosis of CHB patients of DRMS, prognostic markers, or treatment targets.</p>


Subject(s)
Blood Proteins , Metabolism , Case-Control Studies , Female , Hepatitis B, Chronic , Blood , Diagnosis , Humans , Male , Medicine, Chinese Traditional , Proteome , Proteomics
16.
Chinese Journal of Hepatology ; (12): 909-914, 2010.
Article in Chinese | WPRIM | ID: wpr-360799

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the cross-talk between the PI3K/Akt and MEK/ERK pathways and its role in cell cycle regulation under endoplasmic reticulum stress in human hepatocellular carcinoma cells.</p><p><b>METHODS</b>PI3K inhibitor LY294002 and MEK inhibitor U0126 were used to block the PI3K/Akt and MEK/ERK pathways respectively, and constitutively activated Akt mutant construct was used to activate the PI3K/Akt pathway. Western blot was used to study the potential cross-talk between the PI3K/Akt and MEK/ERK pathways under endoplasmic reticulum stress in human hepatocellular carcinoma cells. the role of the cross-talk between the PI3K/Akt and MEK/ERK pathways in cell cycle regulation was investigated by using propidium iodide staining.</p><p><b>RESULTS</b>LY294002 not only blocked Akt activation efficiently but also increased ERK phosphorylation markedly under endoplasmic reticulum stress in SMMC-7721 and Hep3B cells. Furthermore, myr-Akt inhibited endoplasmic reticulum stress-mediated ERK phosphorylation. In contrast, MEK inhibitor U0126 had no effect on endoplasmic reticulum stress-induced Akt activation. It is notable that both myr-Akt overexpression and MEK inhibitor U0126 inhibited endoplasmic reticulum stress-induced G0/G1 phase arrest in SMMC-7721 cells.</p><p><b>CONCLUSION</b>Endoplasmic reticulum stress-induced Akt activation is mediated through PI3K and the PI3K/Akt pathway inactivation is involved in increased ERK activity in human hepatocellular carcinoma cells. The cross-talk between the PI3K/Akt and MEK/ERK cascades plays an important role in endoplasmic reticulum stress-induced human hepatocellular carcinoma cell cycle arrest.</p>


Subject(s)
Butadienes , Pharmacology , Carcinoma, Hepatocellular , Metabolism , Cell Cycle , Cell Line, Tumor , Chromones , Pharmacology , Endoplasmic Reticulum , Metabolism , Extracellular Signal-Regulated MAP Kinases , Metabolism , Humans , Mitogen-Activated Protein Kinase Kinases , Metabolism , Morpholines , Pharmacology , Nitriles , Pharmacology , Phosphatidylinositol 3-Kinase , Metabolism , Phosphorylation , Proto-Oncogene Proteins c-akt , Metabolism , Signal Transduction
17.
Chinese Journal of Hepatology ; (12): 280-283, 2009.
Article in Chinese | WPRIM | ID: wpr-310110

ABSTRACT

<p><b>OBJECTIVE</b>To compare the membrane protein profile of mouse hepatocarcinoma cell H22 with that of normal liver cell.</p><p><b>METHODS</b>The membrane proteins in mouse hepatocarcinoma cell H22 and normal liver cell were extracted and their concentrations were determined by Bradford method. The proteins were separated by two-dimensional electrophoresis, and then stained with silver. The 2-DE maps were scanned and analyzed by Image Master 2D Platinum software. The differential expression protein spots were cut out from the gels, and the peptide fingerprinting was determined by MALDI-TOF-MS (Matrix-Assisted Laser Desorption/ Ionization Time of Flight Mass Spectrometry), followed by matching to Swiss-Prot protein database by Aldente software with experimental pI and MW data.</p><p><b>RESULTS</b>Compared to normal liver cells, 8 membrane proteins, including sulfatase-modifying factor 2, protein kinase C and casein kinase II substrate protein 3, sorting and assembly machinery component 50 homolog, macrophage scavenger receptor types I/II, uncharacterized protein C9 or f135 homolog, tight junction protein ZO-2, 3-hydroxy-3- methylglutaryl-coenzyme A reductase, and vacuolar protein sorting-associated protein 52 homolog were upregulated in H22 cells.</p><p><b>CONCLUSION</b>The membrane proteins involved in cell metabolism, proliferation, signal transduction, and skeleton, which are highly expressed in mouse hepatocarcinoma H22 cells, are probably related to the proliferation, invasion and migration of this tumor cell line.</p>


Subject(s)
Animals , Carcinoma, Hepatocellular , Chemistry , Pathology , Cell Line, Tumor , Electrophoresis, Gel, Two-Dimensional , Hepatocytes , Chemistry , Hydrophobic and Hydrophilic Interactions , Liver Neoplasms , Chemistry , Pathology , Membrane Proteins , Chemistry , Mice , Proteome , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
18.
Chinese Journal of Epidemiology ; (12): 548-551, 2008.
Article in Chinese | WPRIM | ID: wpr-313089

ABSTRACT

Objective To compare the safety and immunogenicity between a MF59-adjuvanted influenza subunit vaccine and a conventional non-adjuvanted influenza subunit vaccine.Methods A randomized, blind-designed controlled study was carried out, with 600 subjects (≥60 years of age)received MF59-adjuvanted influenza subunit vaccine (FLUAD(R), n=400) or conventional non-adjuvanted influenza subunit vaccine (Agrippal(R), n = 200) respectively. The local and systemic reactions were observed on 0-7 days after vaccination. Haemagglutination inhibition (HI) titers of pre-and post-vaccination were detected by the HI assay. Seroconversion (4-fold increase) rate of subjects was calculated using baseline data when it was under naive level, and the protection rate when HI titer achieving the level of protection(≥1:40) after vaccination.Geometric mean titer(GMT) and its increasing folds were calculated.Differences between safety and immunogenicity were also calculated. Results The local and systemic reaction rates were similar between both groups, but the duration in injection site was frequent for Agrippal(R)(P<0.05), while mild pain and fever in injection site were frequent for FLUAD(R). On immunogenicity test, for those subjects whose baseline was under naive level while the seroconversion rate against A/H3N2 viral strain after vaccination - FLUAD(R) was significantly higher than Agrippal(R) (P<0. 001). Aside from A/H1N1 viral strain, the rate of protection on both groups were significantly higher than those from baseline data, but for A/H3N2 viral strain, FLUAD(R) was significantly higher than Agrippal(R) (P<0. 001 ). GMT was higher than baseline (P<0. 001 ) after both groups being vaccinated but FLUAD(R) group was significantly higher than Agrippal(R) group. Conclusion FLUAD(R) was well tolerated by Chinese elderly and its immunogenicity level induced by FLUAD(R) was higher than that of Agrippal(R), showing that it would benefit the elderly with hypoimmunity.

19.
Acta Pharmaceutica Sinica ; (12): 695-699, 2004.
Article in English | WPRIM | ID: wpr-302734

ABSTRACT

<p><b>AIM</b>To compare two methods, the total radioactivity assay (RA method) and the radioactivity assay after separation with high performance liquid chromatography (HPLC-RA method).</p><p><b>METHODS</b>125I-Lidamycin was prepared by Iodogen method and separated by size exclusive high performance liquid chromatography. The pharmacokinetic parameters of lidamycin were assayed by two methods after intravenous injection to mice at the dose of 100 microg x kg(-1), and compared by statistical analysis.</p><p><b>RESULTS</b>The pharmacokinetic parameters (Vd, T1/2alpha, T1/2beta, K21, K10, K12, AUC and CL) showed significant difference between the two methods (P < 0.05).</p><p><b>CONCLUSION</b>The HPLC-RA method was better than the RA method to determine unchanged 125I-lidamycin.</p>


Subject(s)
Aminoglycosides , Blood , Pharmacokinetics , Animals , Antibiotics, Antineoplastic , Blood , Pharmacokinetics , Area Under Curve , Chromatography, High Pressure Liquid , Methods , Enediynes , Female , Iodine Radioisotopes , Isotope Labeling , Male , Mice , Sensitivity and Specificity
20.
Article in Chinese | WPRIM | ID: wpr-266849

ABSTRACT

<p><b>OBJECTIVE</b>To develop the quality control standard of Zhishidaozhi Tabloid Pills.</p><p><b>METHOD</b>Applying TLC to identify Radix et Rhizoma Rhei, Fructus Aurantii Immaturus, Rhizoma Coptidis, Radix Scutellariae, and HPLC to determine the content of emodin of Radix et Rhizoma Rhei.</p><p><b>RESULT</b>Radix et Rhizoma Rhei, Fructus Aurantii Immaturus, Rhizoma Coptidis and Radix Scutellariae could be indentified by TLC. Emodin showed a good linear relationship at a rang of 0.0612-0.612 microgram, r = 0.9999. The average recovery was 97.9%, and RSD was 2.1%.</p><p><b>CONCLUSION</b>The methods are accurate and quick, and can be used for the quality control of Zhishidaozhi Tabloid Pills.</p>


Subject(s)
Chromatography, High Pressure Liquid , Chromatography, Thin Layer , Citrus , Chemistry , Coptis , Chemistry , Drug Combinations , Drugs, Chinese Herbal , Chemistry , Reference Standards , Emodin , Fruit , Chemistry , Plant Roots , Chemistry , Plants, Medicinal , Chemistry , Quality Control , Rheum , Chemistry , Rhizome , Chemistry , Scutellaria , Chemistry
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