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Tang N.,Shaanxi Institute for Food and Drug Control
Zhonghua nan ke xue = National journal of andrology | Year: 2013

To evaluate the safety of intracytoplasmic sperm injection (ICSI) in the mouse model. We simulated clinical ICSI technology and comprehensively evaluated it by parthenogenetic activation, immunofluorescence, embryo transplantation, examination of early implantation, and measurement of the crown-rump length (CRL). ICSI significantly reduced the ability of preimplantation embryo development of the mouse, especially after the 8-cell stage (P < 0.01). The fluorescence of H3K9 dimethylation was abnormal at the male pronuclei of the embryos derived from ICSI. Further examination of the development of the transferred ICSI embryos indicated no significant difference in the rate of early implantation at E5. 5 days as compared with normal fertilization (P = 0.6), but the percentage of "normal embryos" was decreased significantly at E9.5 days (P < 0.01). Obvious growth retardation phenotype was observed even in the normal ICSI embryos at E9.5 days. ICSI might result in growth retardation of embryos by affecting H3K9 dimethylation in the male pronuclei.


Feng B.,Xi'an Jiaotong University | Jin J.,Xi'an Jiaotong University | Wang C.,Shaanxi Institute for Food and Drug Control | Song J.,Xi'an Jiaotong University | And 2 more authors.
Journal of Separation Science | Year: 2012

In order to determine isoflavone glycosides (calycosin-7-O-β-d- glucoside and formononetin-7-O-β-d-glucoside) and aglycones (calycosin and formononetin), a simple HPLC method with isocratic elution employing hydroxypropyl-β-cyclodextrin (HP-β-CD) as a mobile phase additive was developed. Various factors affecting the retention of isoflavone glycosides and aglycones in the C18 reversed-phase column, such as the nature of cyclodextrins, HP-β-CD concentration, and methanol concentration, were systematically studied. The results show that HP-β-CD, as a very effective mobile phase additive, can markedly reduce the retention of isoflavone glycosides and aglycones, and the decrease magnitudes of isoflavone aglycones are more than those of their glycosides. The role of HP-β-CD in the developed HPLC method is attributed to the formation of the inclusion complexes between isoflavone glycosides (or aglycones) and HP-β-CD. So, the apparent formation constants of the isoflavone glycosides (or aglycones)/HP-β-CD inclusion complexes also were investigated. Isoflavone glycosides (and aglycones) form the 1:1 inclusion complexes with HP-β-CD, and the isoflavone aglycones/HP-β-CD complexes are more stable than the isoflavone glycosides/HP-β-CD complexes. Finally, the optimized method was successfully applied for the determination of isoflavone glycosides and aglycones in Radix Astragali samples. © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.


Mao J.,Xi'an Jiaotong University | Mao J.,Affiliated Hospital of Xian Medical College | Liu J.,Xi'an Jiaotong University | Pang X.,Xi'an Jiaotong University | And 5 more authors.
Molecules and Cells | Year: 2012

Atherosclerosis is an inflammatory disease in the vessel wall. Nicotine, a major component of cigarette smoke, is an independent risk factor for cardiovascular diseases including atherosclerosis. As an inflammatory molecule, C- reactive protein (CRP) participates in atherogenesis. Although it has been confirmed that CRP level in smoking patient is significantly higher than non-smokers and cigarette withdrawal,it is unknown whether nicotine induces CRP expression in macrophages. The present study was to observe effect of nicotine on CRP production and the related signal pathway in U937 macrophages. The results showed that nicotine significantly increased mRNA and protein expression of CRP in U937 macrophages in timeand concentration-dependent ways. Nicotinic acetylcholine receptor (nAChR) blocker hexamethonium, MEK1/2 inhibitor PD98059, p38 MAPK inhibitor SB203580 and NF- ?B inhibitor PDTC almost completely abolished nicotineinduced CRP expression in mRNA and protein levels in U937 macrophages. The further study indicated that hexamethonium, PD98059, and SB203580 significantly inhibited ERK1/2 and p38 MAPK phosphorylation. These demonstrate that nicotine has ability to induce CRP expression in macrophages through nAChR-ERK1/2/p38 MAPK-NF-kB signal pathway, which contributes to better understanding of the pro-inflammatory and pro-atherosclerotic effects of nicotine in cigarette smokers. © The Korean Society for Molecular and Cellular Biology.


Pang X.,Xi'an Jiaotong University | Liu J.,Xi'an Jiaotong University | Zhao J.,Xi'an Jiaotong University | Mao J.,The Affiliated Hospital of Xian Medical College | And 6 more authors.
Atherosclerosis | Year: 2014

Objective: Homocysteine (Hcy) is known as an independent risk factor for atherosclerosis. C-reactive protein (CRP) directly participates in initiation and progression of atherosclerosis. However, there is no direct evidence to demonstrate pro-inflammatory effect of Hcy on vascular smooth muscle cells (VSMCs) through CRP. In the present study, we examined the effect of Hcy on CRP expression and investigated the related mechanism in VSMCs. Methods and results: Protein expression and secretion were detected by Western blot and ELISA, respectively. mRNA expression was detected by RT-PCR. Superoxide anion was detected by lucigenin chemiluminometry and the immunofluorescence staining was observed by a fluorescence microscope. The results revealed that Hcy significantly induced mRNA and protein expressions of CRP in VSMCs both invitro and invivo, and anti-IL-1β or anti-IL-6 neutralizing antibody alone or in combination partially reduced Hcy-induced CRP expression. Hcy increased the expression of NR1 subunit of N-methyl- d-aspartate receptor (NMDAr), and MK-801 alleviated Hcy-induced CRP expression in VSMCs. Further studies showed that Hcy-stimulated superoxide anion generation in VSMCs. Nevertheless, pretreatment of the cells with MK-801, TTFA and DPI significantly reduced Hcy-stimulated superoxide anion generation, and antioxidant NAC decreased Hcy-induced CRP expression in VSMCs. Additionally, PD98059, SB205380 or PDTC antagonized Hcy-induced CRP expression, and MK-801, NAC, PD98059 or SB205380 inhibited Hcy-activated phosphorylations of ERK1/2 and p38. Conclusion: The present study demonstrates that Hcy is able to initiate an inflammatory response in VSMCs by stimulating CRP production, which is mediated through NMDAr-ROS-ERK1/2/p38-NF-κB signal pathway. These findings provide new evidence for a role of Hcy in pathogenesis of atherosclerosis. © 2014 Elsevier Ireland Ltd.


Wang C.,Shaanxi Institute for Food and Drug Control | Geng Q.,Shaanxi Institute for Food and Drug Control | Wang Y.,Shaanxi University of Chinese Medicine
Zhongguo Zhongyao Zazhi | Year: 2012

Objective: To study the protective effect of atractylenolide I on immunological liver injury induced by BCG and LPS. Method: Kunming mice were randomly divided into 6 groups: the normal group, the model group, positive control biphenyl group, the atractylenolide I high does group, the atractylenolide I middle dose group and the atractylenolide I low dose group (60, 120, 240 mg·kg -1), with 12 mice in each group. Immunological liver injury in mice was induced by BCG and LPS to compared liver index and spleen index and detect content of serum ALT, AST, MDA and GSH-px in serum and NO, iNOS, TNF-α in serum and liver homogenate. Liver pathological changes were observed by HE staining. Result: Both of atractylenolide I and biphenyl remarkably decrease the increased live index and spleen index (P <0.05), improve the histopathological changes in liver and pathological grades of liver tissues and relieve the inflammatory reaction induced by BCG and LPS. They showed a notable effect in improving MDA and GSH-px in serum. Conclusion: Atractylenolide I can obviously protect immunological injury liver a dose-dependent manner within the range of test doses. Its mechanism may be related to release or over expression of inhibitory inflammatory medium such as NO, iNOS and TNF-α.


Wang C.,Shaanxi Institute for Food and Drug Control | Liu H.,Shaanxi Institute for Food and Drug Control | Zhang B.,Shaanxi Institute for Food and Drug Control | Guo H.,Shaanxi Institute for Food and Drug Control
Journal of Separation Science | Year: 2011

To separate and determine oleanolic acid and ursolic acid, a rapid and accurate HPLC using γ-CD as the mobile phase additive was developed. The effect of CD nature and concentration, and the acidity of the mobile phase on the chromatographic behavior of two bioactive triterpenes were systematically studied. Two bioactive triterpenes were completely separated (R = 3.11) on a Kromasils C18 column (150 × 4.6mm id, 5 μm) with the mobile phase consisting of acetonitrile/0.1% phosphoric acid with 2mM γ-CD as the mobile phase modifier (60:40, v/v). The flow rate was set at 1.0 mL/min and the eluent was detected at 210 nm for two bioactive triterpenes. The linearity of the method was excellent (r=0.9999) over the studied range of 6-300 μg/mL for oleanolic acid, and 12-600 μg/mL for ursolic acid. The LOD and LOQ were 1.5 and 5.0, 1.0 and 3.0 μg/mL for oleanolic acid and ursolic acid, respectively. The optimized method was successfully applied to separate and determine two bioactive triterpenes in five Chinese herbs. It is concluded that this method could be used for rapid and accurate qualitative and quantitative analysis of the two bioactive triterpenes in Chinese herbs. © 2011 WILEY-VCH Verlag GmbH &Co. KGaA, Weinheim.


Wang C.-H.,Shaanxi Institute for Food and Drug Control | Wang Y.-X.,Shaanxi University of Chinese Medicine | Liu H.-J.,Shaanxi Institute for Food and Drug Control
Journal of Pharmaceutical Analysis | Year: 2011

A simple, precise, and rapid high-performance liquid chromatographic method was developed and validated for the simultaneous determination of vitexin-2″-O-glucoside, vitexin-2″-O-rhamnoside, rutin, vitexin, and hyperoside. The HPLC separation was performed using a Shim-pack VP-ODS C 18 column (250 mm×4.6 mm i.d., 5 μm) with the isocratic mobile phase consisting of tetrahydrofuran/ acetonitrile/0.05% phosphoric acid solution (20:3:77, v/v/v), and the flow rate was set at 1.0 mL/min. UV detection was carried out at a wavelength of 360 nm and the whole analysis took 25 min. The method was linear in the range of 4.12206.00 μg/mL for vitexin-2″-O-glucoside, 4.05202.50 μg/mL for vitexin-2″-O- rhamnoside, 1.6482.00 μg/mL for rutin, 1.7487.00 μg/mL for vitexin, and 1.4170.60 μg/mL for hyperoside with the correlation coefficient for each analyte more than 0.998. The limit of detection (LOD) and limit of quantitation (LOQ) were 0.6 and 2 ng for vitexin-2″-O-glucoside, 0.6 and 2 ng for vitexin-2″-O-rhamnoside, 0.3 and 1 ng for rutin, 1 and 3 ng for vitexin, and 0.5 and 2 ng for hyperoside, respectively. Intra- and inter-day precision and accuracy (RSD) were less than 3%. The developed HPLC method was successfully applied to the analysis of five flavonoids in hawthorn leaves, hawthorn fruits, and the preparations containing hawthorn leaves or fruits. © 2011 Xi'an Jiaotong University. Production and hosting by Elsevier B.V. All rights reserved.


Li M.,Xi'an Jiaotong University | Li M.,Lianyungang Hospital of Traditional Chinese Medicine | Liu J.,Xi'an Jiaotong University | Han C.,Xi'an Jiaotong University | And 4 more authors.
Cellular Physiology and Biochemistry | Year: 2011

Atherosclerosis is an inflammatory disease in the vessel wall. As an inflammatory molecule, C-reactive protein (CRP) participates in all stages of atherosclerotic process. Although angiotensin II (Ang II) can stimulate the vascular cells to produce CRP, it is unknown whether Ang II induces CRP expression in macrophages. The present study was to observe effect of Ang II on CRP production and the related signal pathway in U937 macrophages so as to provide more evidence for the proinflammatory action of Ang II. The results showed that Ang II significantly increased mRNA and protein expression of CRP in U937 macrophages in time- and concentration-dependent manners. AT1 receptor blocker losartan blocked Ang II -induced CRP expression in mRNA and protein levels in U937 macrophages. Losartan and complex II inhibitor TIFA decreased Ang II -stimulated reactive oxygen species (ROS) generation, and antioxidant NAC completely abolished Ang II -induced CRP expression in U937 macrophages. The further study indicated that losartan, NAC, MEK1/2 inhibitor PD98059, p38MAPK inhibitor SB203580 obviously inhibited ERK1/2 and p38MAPK phosphorylation, and PD98059, SB203580 and NF-κB inhibitor PDTC reduced Ang II -induced mRNA and protein expression of CRP in U937 macrophages. These demonstrate that Ang II is capable of inducing CRP generation in macrophages via AT1-ROS-ERK1/2/p38MAPK-NF-κB signal pathway, which contributes to better understanding of the proinflammatory and proatherosclerotic actions of Ang II. © 2011 S. Karger AG, Basel.


Zeng A.,Xi'an Jiaotong University | Xing J.,Xi'an Jiaotong University | Wang C.,Shaanxi Institute for Food and Drug Control | Song J.,Xi'an Jiaotong University | And 3 more authors.
Analytica Chimica Acta | Year: 2012

In order to differentiate two species of Radix Puerariae (Radix Puerariae lobatae and Radix Puerariae thomsonii) and to determine major isoflavonoids (puerarin, daidzin, daidzein and genistein) in the samples, a simple high performance liquid chromatography (HPLC) method with isocratic elution employing cyclodextrins (CDs) as mobile phase additives was developed. Various factors affecting the retention of isoflavonoids in the C18 reversed-phase column, such as the nature of CDs, the concentration of hydroxypropyl-β-cyclodextrin (HP-β-CD) and the methanol percentage in the mobile phase, were studied. Experimental results confirmed that HP-β-CD, as a very effective mobile phase additive, could markedly reduce the retention of isoflavonoids, especially daidzein and genistein. The elution of four isoflavonoids could be achieved on a Kromasil® C18 column within 56min by using the methanol-water contained 5mM HP-β-CD (25/75, v/v) mixture as the mobile phase. The formation of the inclusion complexes between isoflavonoids and HP-β-CD explained the modification of the retention of analytes. The apparent formation constants determined by HPLC confirmed that the stoichiometry of HP-β-CD-isoflavonoid complexes was 1:1, and the stability of the complexes depended on the size and property of isoflavonoids. The optimized method was successfully applied for the simultaneous determination of major isoflavonoids in P. lobatae and P. thomsonii samples. This work provides a useful method for the analysis of traditional Chinese herbs. © 2011 Elsevier B.V.


PubMed | Shaanxi Institute for Food and Drug Control and Xi'an Jiaotong University
Type: Journal Article | Journal: Journal of chromatographic science | Year: 2017

In this work, a new molecularly imprinted solid phase extraction protocol was developed for the selective extraction and purification of glycyrrhizic acid from liquorice roots in aqueous media. The molecularly imprinted polymers (MIPs) for glycyrrhizic acid were prepared by using bismethacryloyl--cyclodextrin and methacrylic acid as double functional monomers and characterized by Fourier transform infrared spectroscopy, scanning electron microscope, thermo gravimetric analysis, nitrogen adsorption and elemental analysis. In aqueous media, the adsorption properties of MIPs including adsorption kinetics, adsorption isotherms and selectivity adsorption were investigated. The characterization of imprinted polymers indicated that the prepared MIPs had good stability and many cavity structures. The results of adsorption experiments illustrated the MIPs had high adsorption capacity of glycyrrhizic acid (69.3 mg g

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