The antitussive and expectorant mechanism of Platycodon total saponins based on metabonomics / 药学学报

UHPLC-Q-TOF/MS metabonomics technology was used to clarify the metabolic regulation pathways by which Platycodon total saponins (PTS) exert antitussive and expectorant effects in a mouse cough model, in which coughing is induced by concentrated ammonia, and in a phenol red excretion model. After approval by the Experimental Animal Ethics Committee of Jiangxi University of Chinese Medicine (Approval No. JZLLSC-20190235), the mice were randomly divided into a normal group, a model group, a positive drug group and a PTS group. Endogenous metabolites in mouse serum were identified by UHPLC-Q-TOF/MS. Principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) were used for multivariate analysis. Metabolic pathways were analyzed by the Metaboanalyst platform. The results show that PTS can significantly prolong the cough latent period and cough frequency of mice, and significantly increase phenol red excretion. UHPLC-Q-TOF/MS identified 19 metabolites related to cough, and PTS significantly decreased 16 of them; 17 metabolites related to expectoration were identified, and PTS decreased the levels of all. Metabolic pathway analysis showed that linoleic acid metabolism, arachidonic acid metabolism and glycerophospholipid metabolism were the main pathways involved in serum metabolite changes in this mouse cough model. Linoleic acid metabolism, phenylalanine, tyrosine and tryptophan biosynthesis, arachidonic acid metabolism, phenylalanine metabolism and α-linolenic acid metabolism were the main pathways involved in serum metabolite changes in the phenol red excretion model. This study is the first to elucidate the regulation of antitussive and expectorant metabolic pathways and the effect of PTS on these pathways.

Mechanism of Banxia Xiexintang on Rats with Chronic Gastritis Based on Quantitative Proteomics / 中国实验方剂学杂志

Objective:To study the biological basis of Banxia Xiexintang against chronic gastritis by using quantitative proteomics. Method:The experimental rats were divided into normal group，chronic gastritis model group，and Banxia Xiexintang group. The chronic gastritis model was established four weeks later by gavage with 56% ethanol. After treatment，the stomach tissues were stained with hematoxylin and eosin （HE） to observe the histopathological damage and improvement of gastric tissue in each group. The protein in gastric tissue was extracted. The differential proteins among different groups were studied by quantitative proteomics using tandem mass spectrometry tag（TMT），and the key differentially expressed proteins（DEPs） were verified by Western blot. Result:A total of 4 452 proteins were identified from rat stomach tissues，of which 318 proteins were different between the model and the normal group. After the intervention of Banxia Xiexintang，compared with the model group，there were a total of 258 differential proteins， which were mainly enriched in cell killing，nucleoid and hijacked molecular function. Kyoto encyclopedia of genes and genomes（KEGG） pathway enrichment included tricarboxylic acid（TCA） cycle，steroid hormone biosyntheis，and Retrograde endocannabinoid signaling，as well as phosphatidylinositol 3-kinase（PI3K）/protein kinase B（Akt），nuclear transcription factor-<italic>κ</italic>B（NF-<italic>κ</italic>B） signal pathways. Western blot verification found that 14-3-3 theta，Tenascin-C，ntercellular adhesion molecule-1（ICAM-1），stem cell factor（SCF），Caspase-3，GTPase of the Immunity-associated protein 4（GIMAP4） and mitochondrial pyruvate carrier 1（Mpc1） might be the crucial proteins for the treatment of chronic gastritis. Conclusion:The mechanism of Banxia Xiexintang in the treatment of chronic gastritis involves energy metabolism，hormone regulation，inflammation and immune processes. The target proteins found by differential proteomics and the signaling pathways may be the biological basis of Banxia Xiexintang in the treatment of chronic gastritis.

Revealing the synergistic mechanism of Shenfu Decoction for anti-heart failure through network pharmacology strategy / 中国天然药物

The present study was designed to investigate the targets and synergistic mechanism of Shenfu Decoction (SFD) in the treatment of heart failure. A heart failure animal models was established to evaluate the pharmacological effects of SFD for anti-heart failure, then constructed ingredient-target interaction network by developing ingredient and target databases, the Discovery sdudio software was used for molecular docking. In addition, we validated the predicted protein targets of active ingredients in SFD by using surface plasmon resonance (SPR) technology. Our results demonstrated that SFD could enhance ejection fraction, alleviate myocardial histopathological characteristics, and reduce the level of angiotensin converting enzyme (ACE), aldosterone (ALD), atrial natriuretic polypeptide (ANP) and Renin (REN) in heart failure rat model. In addition, the ingredient database including 349 constituents and target database including 236 proteins were established, and 75 proteins were screened and identified by molecular docking strategy. 22 core target proteins were identified through network pharmacology, and the component-core target network was constructed. Finally, the affinity between the compounds and targets were verified by the SPR analysis method. The present study suggested that SFD may act on ACE 2, REN, ACE, ICAM-1, EGF, HTR2B, PARP1, NPPB and other proteins through AC, BAC, ACN, Re, Rg1, Rb1 to exert synergistic effects against heart failure.

Tissue distribution of TPGS modified artesunate liposome and its metabolites in rats / 中国中药杂志

Artesunate, which is a widely used anti-malaria medicine, can be made into liposome to overcome its poor bioactivity. Its tissue distribution in rats may change with different dosage forms, which therefore shall be studied after ARS-TPGS-Lipo was injected. Based on this experiment, ARS-TPGS-Lipo and ARS-Lipo were prepared by thin-film hydration method. LC-MS/MS method was used to simultaneously determine ARS and DHA in rat tissues at different time points. The results showed that this method was suitable for the content analysis of ARS and DHA in biological samples. The distribution of ARS and DHA in ARS-TPGS-Lipo, ARS-Lipo and ARS groups were quite different. The content of ARS-TPGS-Lipo in liver was the highest, with significant differences.ARS and DHA contents in ARS group eliminated rapidly. ARS and DHA contents in ARS-Lipo group were higher in liver and spleen, while those in ARS-TPGS-Lipo group significantly increased only in liver (<0.05).

Feasibility study of QAMS for quantitative analysis of multiple structural types of ingredients in Atractylodis Rhizome by GC / 中国中药杂志

In this study， quantitative analysis of multi-components with single marker(QAMS) was established and validated to simultaneously determine four sesquiterpenoids(β-eudesmol， atractylon， atractylolideⅠ， atractylolide Ⅱ) in Atractylodis Rhizome based on the gas chromatographic method(GC). Using β-eudesmol as the contrast， the relative correctionfactors(RCF) of the other three sesquiterpenoids were determined by GC. Within the line arranges，the values of RCF of β-eudesmol to atractylon， atractylolideⅠand atractylolide Ⅱ were 0.823， 0.690 and 0.766， respectively. The RCF had a good reproducibility in various instruments， chromatographic columns. According to their RCF， we simultaneously determined four sesquiterpenoids in Atractylodis Rhizome only using one marker. The results of QAMS method were validated by comparing with that of internal standard method， and no obvious significant difference was found.

Simultaneous determination of six constituents in different kinds of Atractylodes rhizome by GC / 中成药

AIM To establish a GC method for the simultaneous determination of β-eudesmol,atractylon,atractylodin,atractylolide Ⅰ,atractylaxanthin Ⅱ and (4E,6E,12E)-tetradecene-8,10-diyne-1,3-diacetate in Atractylodes rhizome,and cluster analysis of A.rhizome according to the content level.METHODS The analysis of A.rhizome solution was performed on an HP-5 capillary column (30 m × 0.32 mm,0.25 μm) with FID as the detector,the initial temperature 100 ℃,with 10 ℃/min to 135 ℃;with 1 ℃/min to 150 ℃;with 50 ℃/min to 200 ℃ (keeping 6 minutes),and with 50 ℃/min to 250 ℃ (keeping 8 minutes).The FID detector temperature was 300 ℃ and the injector temperature was 250 ℃,with the flow rate carrier gas 1.4 L/min;The tail gas was N2 (99.999％),with the ratio of carrier gas Air ∶ H2 ∶ N2 =400 ∶ 30 ∶ 25;The sample volume was 1 μL,and the split ratio was 20 ∶ 1.The results were analyzed by cluster analysis with SPSS 21.0 statistical software.RESULTS Six constituents showed good linear relationships within their own ranges (r ＞ 0.999 6),whose average recoveries were 99.46％-100.95％ with the RSDs of 0.09-0.41.A.rhizome was divided into three categories.CONCLUSION This accurate,stable and reproducible method can be used for the quality control of A.rhizome.

Simultaneous determination of eleven constituents in Huxin Oral Liquid by LC-MS/MS / 中成药

AIM To establish a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the simultaneous content determination of matrine,oxymatrine,salvia acid B,tanshinol,protocatechualdehyde,liquiritin,astragaloside,cinnamaldehyde,tanshinone Ⅱ A,ophiopogonin D and ruscogenin in Huxin Oral Liquid (Glycyrrhizae Radix et Rhizoma,Astragali Radix,Ophiopogonis Radix,etc.).METHODS The analysis of methanol extract of this drug was performed on a 25 ℃ thermostatic Ultimate XB-C18 column (4.6 mm × 100 mm,3 μm),with the mobile phase comprising of water-methanol (containing 0.05％ formic acid) flowing at 0.4 mL/min in a gradient elution manner.RESULTS Eleven constituents showed good linear relationships with-in their own ranges (r ＞0.999 0),whose average recoveries were 96.66％-103.2％ with the RSDs of 1.4％-3.4％.CONCLUSION This sensitive,reliable and accurate method can be used for the quality control of Huxin Oral Liquid.

Simultaneous determination of four constituents in Yangxin Dingji Capsules by HPLC / 中成药

AIM To establish an HPLC method for the simultaneous content determination of four constituents in Yangxin Dingji Capsules (a cardiac tonic for palpitation,containing Glycyrrhizae Radix et Rhizoma Praeparata cum Melle,Cinnamomi Ramulus,Rehmanniae Radix,etc.).METHODS The analysis of 50％ methanol extract of Yangxin Dingji Capsules was performed on a 40 ℃ thermostatic Diamonsil C1s column (250 mm × 4.6 mm,5 μm),with the mobile phase comprising of 0.1％ formic acid-acetonitrile flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 265 nm.RESULTS Liquiritin,glycyrrhizic acid,cinnamic acid and cinnamic aldehyde showed good linear relationships within the ranges of 1.00-80.24 μg/mL (r=0.999 0),2.52-100.70 μg/mL (r--0.999 7),0.50-40.40 μg/mL (r =1.000 0) and 0.66-52.96 μg/mL (r =1.000 0),whose average recoveries were 97.74％,100.97％,101.48％ and 99.49％ with the RSDs of 0.45％,1.11％,1.27％ and 1.66％,respectively.CONCLUSION This simple,accurate and reproducible method can be used for the quality control of Yangxin Dingji Capsules.

Improvement of kidney yang syndrome by icariin through regulating hypothalamus-pituitary-adrenal axis / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate whether Epimedium brevicornu Maxim (EB) and icariin could exert their protective effects on hydrocortisone induced (HCI) rats by regulating the hypothalamus-pituitary-adrenal (HPA) axis and endocrine system and the possible mechanism.</p><p><b>METHODS</b>Male 10-week-old Sprague Dawley (SD) rats were allotted to 6 groups (A-F) with 12 each, group A was injected normal saline (NS) 3 mL/kg day intraperitoneally, group A and B were given NS 6 mL/kg day by gastrogavage, group B-F were injected hydrocortisone 15 mg/kg intraperitoneally, group C and D were given EB 8 or 5 g/(kg day) by gastrogavage, group E and F were given icariin 25 or 50 mg/(kg day) by gastrogavage. Gene expressions of hypothalamus corticotropin releasing hormone (CRH) and pituitary proopiomelanocortin (POMC) were detected by reverse transcription-polymerase chain reaction (RT-PCR), and protein of pituitary POMC by Western-blot.</p><p><b>RESULTS</b>The serum T4, testosterone, cortisol and POMC mRNA expression were increased after treatment with EB or icariin in HCI rats, the serum CRH and the hypothalamus CRH mRNA expression released from hypothalamus corticotropin decreased compared with group B (P<0.05).The treatment with only icariin increased serum adrenocorticotropic hormone (ACTH) compared with group B (P<0.05).</p><p><b>CONCLUSION</b>EB and icariin might be therapeutically beneficial in the treatment of HCI rats through attuning the HPA axis and endocrine system which was involved in the release of CRH in hypothalamic, and the production of POMC-derived peptide ACTH in anterior pituitary, the secretion of corticosteroids in adrenal cortex.</p>

Effect of Gegen Qinlian decoction on hepatic cytochrome CYP450 isozymes in rats by HPLC-MS/MS / 中国中药杂志

To study the effect of Gegen Qinlian decoction and its major effective components on five hepatic microsomal CYP450 isozymes in rats. The in vitro hepatic microsomal incubation technique was used to co-culture Gegen Qinlian decoction and its major effective components together with each probe substrate. HPLC-MS/MS was used to establish the analytical method for metabolites of the five isoform probe substrates of CYP450 isozymes, detect the linearity among micoromal protein concentration, incubation time and metabolite formation amount. And HPLC-MS/MS was applied to determine the formation rate (V) of corresponding metabolites (acetaminophen, 4-OH-chlorzoxazone, dextrophan, 6-OH-chlorzoxazone and 6β-hydroxytestosterone) specific probe substrates of the five isoform probe substrates of CYP450 isozymes (phenacetin, polbutamide, dextromethorphan, chlorzoxazone, testosterone), in order to determine the activity of each isozyme. The result showed good linearity among acetaminophen, 4-OH-tolbutamide, dextrophan, 6-OH-chlorzoxazone and 6β-hydroxytestosterone, satisfactory precision, stability and average recovery, suggesting the method was feasible. The optimized in vitro microsomal incubation conditions conformed to the requirements in the guideline of drug-drug interaction. Gegen Qinlian decoction showed different degrees of inhibitor effect on 5 CYP450 isoforms (CYP1A2, CYP2C11, CYP2D2, CYP2E1, CYP3A1/2). Its major effective component berberine could inhibit each CYP450 isoform at high concentrations (except for CYP1A2, CYP3A1/2).

Analysis and comparison of intestinal absorption of components of Gegenqinlian decoction in different combinations based on pharmacokinetic parameters / 药学学报

To analyse and compare the characteristics of the intestinal absorption of puerarin, baicalin, berberine and liquiritin in different combinations of Gegenqinlian decoction based on pharmacokinetic parameters, a sensitive liquid chromatography-tandem mass spectrometric (LC-MS/MS) method was applied for the quantification of four components in rat's plasma. And pharmacokinetic parameters were determined from the plasma concentration-time data with the DAS software package. The influence of different combinations on pharmacokinetics of four components was studied to analyse and compare the absorption difference of four components, together with the results of the in vitro everted gut model and the rat single pass intestinal perfusion model. The results showed that compared with other combinations, the AUC values of puerarin, baicalin and berberine were increased significantly in Gegenqinlian decoction group, while the AUC value of liquiritin was reduced. Moreover, the absorption of four components was increased significantly supported by the results from the in vitro everted gut model and the rat single pass intestinal perfusion model, which indicated that the Gegenqinlian decoction may promote the absorption of four components and accelerate the metabolism of liquiritin by the cytochrome P450.

Study on effect of different compatibility of saponins contained in banxia xiexin tang on intestinal bacterium metabolism / 中国中药杂志

<p><b>OBJECTIVE</b>To compare the effect of intestinal flora on the metabolism of the Banxia Xiexin Tang (BXT) full prescription group, the sweet-nourishing group and saponins contained in single ingredients ginseng and liquorices.</p><p><b>METHOD</b>The anaerobic incubation technology for intestinal flora in vitro was adopted to incubate the BXT full prescription group, the sweet-nourishing group and extracting solution of the single ingredients, under anaerobic conditions at 37 degrees C. Samples of different incubating time points were collected. The high-speed separation and content determination of various prototypes and metabolites were conducted with LC-MS/MS method, and then their degradation rate K was calculated to observe the difference and characteristics in metabolism of different compatible groups.</p><p><b>RESULT</b>Intestinal flora could transform saponins into their metabolites. Having comparing spss one factor variance, we learned the difference in saponin metabolites of different compatible groups. As for the degradation rate of glycyrrhizic acid, the sweet-nourishing group > the full prescription group > the single prescription group (P < 0.05). Rb1 degraded the most slowly in the full prescription group. As for the degradation rate of Re, the single prescription group > the sweet-nourishing group > the full prescription group (P < 0.05).</p><p><b>CONCLUSION</b>The sweet-nourishing group and the sweet-nourishing group have different effect in inducing or inhibiting intestinal flora. The single prescription group shows in inhibition in metabolites of Rb1 and Rg1. Glycyrrhizic acid metabolites are promoted by glycyrrhetinic acid, which facilitates the efficacy of drug absorption. The compatibility of compounds has no impact on metabolites of Rb1 and Rg3.</p>

A new anthraquinone glycoside from the root of Rheum officinale Baill / 药学学报

A new and a known anthraquinone glycosides were isolated from the ethanol extract of the roots of Rheum officinale Baill. The extract was purified by various chromatographies, such as silica gel, Sephadex LH-20, RP-C18 column chromatography and HPLC. Two compounds were identified by the spectroscopic techniques of NMR, MS, and chemical method. In addition, they were tested for their cytotoxic effects against HepG2 cell. Unfortunately, they showed no or weak activity.

Intestinal absorption of different combinations of active compounds from Gegenqinlian decoction by rat single pass intestinal perfusion in situ / 药学学报

The aim is to study the intestinal absorption of different combinations of active compounds out of Gegenqinlian decoction. Rat single pass intestinal perfusion model with jugular vein cannulated was used. Samples were obtained continuously from the outlet perfusate and the mesenteric vein. The levels of puerarin, daidzin, liquilitin, baicalin, wogonoside, jatrorrhizine, berberine and palmatine were determined by LC-MS/MS and their permeability coefficients were calculated. The results showed that Glycyrrhiza could promote the absorption of the active ingredients in Pueraria which is the monarch herb; meanwhile, Pueraria also played a role in promoting the absorption of liquilitin. Based on the Gegenqinlian decoction and the different combinations experiments, the results concerning the absorption of baicalin and wogonoside were as follows. For baicalin, Pueraria and Glycyrrhiza could promote its absorption and the effect of Pueraria was more obvious. For wogonoside, Pueraria could also promote its absorption, while Glycyrrhiza played a opposite role. Pueraria and Glycyrrhiza both played a part in promoting the absorption of jateorhizine, berberine and palmatine, the effective compounds in Coptis.

Wnt/b-catenin signaling pathway affects the protein expressions of caspase-3, XIAP and Grp-78 in hepatocellular carcinoma / 中华肝脏病杂志

To investigate the relationship and significance of Wnt/b-catenin signaling pathway with caspase-3, XIAP, HSP27and Grp-78. The HCC cell line HepG2 was transfected with small interfering RNA (siRNA) directed against b-catenin. After 72 and 96 h, protein was extracted and the protein expressions of b-catenin, caspase-3, XIAP, Grp-78 and HSP27 were detected by Western blot. b-catenin protein expression was inhibited at both time points and the expression at 96 h was higher than that at 72 h (F = 160.72, P is less than to 0.01). Interestingly, Caspase-3 protein expression was decreased at 72 h and increased to normal at 96 h (F = 136.10, P is less than to 0.01), while p-caspase-3 protein expression increased at 72 h and decreased to normal at 96 h (F = 98.65, P is less than to 0.01). XIAP protein expression decreased at 72 h (F = 37.29, P is less than to 0.01) and increased at 96 h. Grp-78 protein expression increased at 72 h and decreased to normal at 96 h ( F = 58.72, P is less than to 0.01). HSP27 protein expression showed no change following transfection ( F = 1.91, P is more than to 0.05). Wnt/b-catenin signaling pathway is related to the protein expressions of caspase-3, XIAP and Grp-78, but not related to HSP27 protein expression in HCC. Wnt/b-catenin signaling pathway may participate in the regulation of HCC apoptosis, proliferation and differentiation through affecting these factors.

Molecular analysis of TP53, D9S1747, D9S162 and RPS6 in oral squamous cell carcinoma / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the correlation between the frequency of molecular abnormalities of 4 loci at chromosomal 9p21 (D9S1747, D9S162, RPS6) and 17p13 (TP53) and the clinical characteristics and prognosis.</p><p><b>METHODS</b>The oral squamous cell carcinoma (OSCC) lesions in 71 patients were manually microdessected. Genomic DNA from these lesions and normal lymphnode tissu or peripheral blood of the same patients were extracted using the Watson's tissue kit. The loss of heterozygosity (LOH) and microsatellite instability (MI) of 17p13 and 9p21 were analyzed by PCR-page electrophoresis after DNA extraction.</p><p><b>RESULTS</b>LOH and MI were detected in the OSCC of 48 patients (68%). The LOH and MI frequency at chromosomes 17p13 and 9P21 were 56% (35/63) and 59% (40/68) respectively. The LOH and MI frequency at 9p21 was significantly associated with WHO grading (P < 0.01) and lymphonode metastasis (P < 0.01). The LOH and MI frequency at 17p13 was significantly associated with clinical stage (P < 0.05). TP53 genetic aberration and 9p21 genetic aberration were significant prognostic factors for OSCC. The prognosis was poor in the LOH and MI positive group of chromosome 17p13 and 9p21. The frequency of LOH and MI at TP53 was the only independent factor for overall survival (P < 0.05).</p><p><b>CONCLUSIONS</b>The LOH and MI of 17p13 and 9p21 were related to clinical stage and lymphonode metastasis. LOH of TP53 was an independent prognostic factor for OSCC.</p>

Effect of laser surface hardening technology on the corrosion resistance of dental casting alloy / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the effect of the laser surface hardening technology on the corrosion resistance of dental casting alloy.</p><p><b>METHODS</b>Twenty-three cobalt-chromium alloy specimens were made in this study. Twenty-two specimens were equally divided into two groups randomly. One was experimental group for laser surface hardening processing and the other was control group without any treatment. In each group, ten specimens were used for corrosion analysis by electrochemical method, and one for surface metallographical structure and morphology observation by scanning electron microscope. Remaining one specimen was partially processed on limited area for surface metallographical structure and morphology comparison.</p><p><b>RESULTS</b>Metal grains distributed uniformly and achieved a good refinement with mainly the same size in experimental group. Metal grains in specimen which processed in its partial surface area also achieved a good refinement in the laser processing area. There was statistical difference in electric potential of corrosion and logarithmic value of current of corrosion between experimental group and control group (P < 0.01).</p><p><b>CONCLUSION</b>Laser surface hardening technology has a positive effect in improving the corrosion resistance of dental casting alloy in artificial saliva.</p>

The role and significance of Wnt/beta-catenin signaling pathway regulating the signaling molecules in hepatocellular carcinoma / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the role and significance of Wnt/beta-catenin signaling pathway regulating GSK-3beta, STAT3, Smad3 and TERT in hepatocellular carcinoma (HCC).</p><p><b>METHODS</b>The HCC cell line HepG2 was transfected with small interfering RNA (siRNA) directed against beta-catenin. Proteins were extracted and the expressions of beta-catenin, GSK-3beta, p-GSK-3beta, STAT3, Smad3 and TERT were detected by Western blot at 72 h and 96 h respectively after transfection.</p><p><b>RESULTS</b>beta-catenin expression was inhibited at both time points and the expression at 96 h was higher than that at 72 h (t = 4.43, P < 0.05). Interestingly, GSK-3beta and p-GSK-3beta expressions increased gradually at 72 and 96 h (tGSK-3beta= 4.98, tp-GSK-3beta= 29.83, P < 0.05) respectively, and STAT3 expression showed no alteration after transfection (F = 0.49, P > 0.05). Smad3 expression was increased at 72 h (t = 10.67, P < 0.05) and decreased to normal at 96 h (t = 1.26, P < 0.05), while TERT expression decreased at 72 h (t = 4.18, P is less than 0.05) and increased to normal at 96 h (t = 1.26, P > 0.05).</p><p><b>CONCLUSIONS</b>Wnt/beta-catenin signaling pathway is related to the expressions of GSK-3beta, Smad3 and TERT, but perhaps not related to STAT3 protein expression in HCC. It suggested that Wnt/beta-catenin signaling pathway might participate in HCC genesis and development through regulating the above three factors.</p>

Roles of cyclooxygenase-2 in microvascular endothelial cell proliferation induced by basic fibroblast growth factor / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The level of basic fibroblast growth factor (bFGF) increases rapidly after cerebral ischemia. However, the molecular mechanisms for the effects of bFGF on cerebral microvascular endothelial cells (cMVECs) have not yet been fully elucidated. In this study, a murine cMVEC line, bEnd.3, was employed to study the effects of bFGF on cyclooxygenase (COX) expression and its downstream effects in cMVECs.</p><p><b>METHODS</b>After treatment with bFGF, RT-PCR and Western blotting analyses were carried out to evaluate the changes in COX-2 mRNA and protein expression, respectively. MTT assays were performed to measure cell proliferation. The prostaglandin E2 (PGE2) and vascular endothelial growth factor (VEGF) concentrations in the culture medium were measured by enzyme-linked immunosorbent assay (ELISA).</p><p><b>RESULTS</b>COX-2 mRNA and protein expressions in bEnd.3 cells were induced by bFGF in time- and dose-dependent manners. The bFGF-induced COX-2 upregulation led to enhanced PGE2 production by bEnd.3 cells, and this effect was abolished by the selective COX-2 inhibitor NS-398. bFGF also increased VEGF production by bEnd.3 cells, and this effect was blocked by NS-398 and the EP1/2 (PGE2 receptors) antagonist AH6809. Furthermore, exogenous PGE2 increased VEGF production in bEnd.3 cells, and AH6809 blocked this effect.</p><p><b>CONCLUSION</b>bFGF increases VEGF production in an autocrine manner by increasing COX-2-generated PGE2 in cMVECs and subsequently stimulates MVEC proliferation and angiogenesis.</p>

The TP53 and RPS6 alterations at the invasive tumor front, center and stroma of oral squamous cell carcinoma / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To assess the difference of genetic alteration patterns among different areas in the same oral squamous cell carcinoma (OSCC).</p><p><b>METHODS</b>Studied the loss of heterozygosity (LOH) and microsatellite instability (MI) at chromosomal loci TP53 and RPS6 on the invasive tumor front (ITF), the center/superficial part and stroma cells by combining laser capture microdissection (LCM) and PCR technique.</p><p><b>RESULTS</b>There existed a high frequency of LOH and MI on chromosomes loci TP53 and RPS6. The frequency of RPS6 and TP53 aberration at the stroma was 23.5% (4/17) and 43.8% (7/16), respectively. While in epithelial part (both ITF and center), it reached up to 64.7% (11/17) and 70.6% (12/17) respectively, and the difference was significant (P < 0.05). The overall frequency of the two markers was statistically higher at the ITF (20/32) than at the center/superficial part (15/34) (P < 0.05).</p><p><b>CONCLUSIONS</b>The current study revealed that genetic alterations were different in different areas of the same tumor and there existed a relationship between the histological grading and genotypes of OSCC.</p>