The UHPLC-LTQ-Orbitrap high resolution mass spectrometer was used to explore the chemical compositions in safflower. The rapid separation of the compositions was conducted by the UHPLC, following by high resolution full scan and MS2 scan, under the positive and negative ion mode. The chemical formula of compositions were deduced by full scan data in less than 5, then the potential structures were confirmed by the MS2 data. Forty-nine compounds were detected, of which 26 was identified, and 5 compounds was validated by the standard substances.
Carthamus tinctorius ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Molecular Structure ; Tandem Mass Spectrometry ; methods

Animals ; Esophagus ; metabolism ; Glucagon-Like Peptide 2 ; pharmacology ; Intestinal Mucosa ; drug effects ; metabolism ; Intestine, Small ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Stomach ; metabolism

Objective To analyze the causes associated with the failure of dental implant restoration. Methods The patients who received dental implant restoration from January 2001 to December 2008 in Center of Dental Implant, School of Stomatology, Shandong University were reviewed and analyzed.The cases with implant loosening, broken or removed were considered failure. Results There were a total of 38 failure implants in 32 patients found in this group of patients. Of those, 33 implants loosened (17 cases before restoration and 16 cases after restoration), two were broken, two retention screws broken and one implant perforated on buccal side. The causes of failure included doctor-related factors in 19 cases, patientrelated factors in 9 cases, implant-related factors in two cases and two uncertainties. Conclusions Doctorrelated factor is the main cause of dental implant failure, followed by patient-related factor and implantrelated factor.

Dioscin has a wide range of biological effects and broad application prospects. However the studies concerning the toxicology and mechanism of dioscin is small. This article is to study the hepatotoxicity of dioscin and the effect of dioscin treatment on expression of aryl hydrocarbon receptor (AhR) mRNA and CYP1A mRNA and protein in HepG2 cells in vitro. Dioscin 0.5-32 µmol · L(-1) exposed to HepG2 cells for 12 h, cell viability was examined by CCK-8 assay and the release rate of lactate dehydrogenase (LDH) was to evaluate cell membrane damage. HepG2 cells morphologic changes were quantified by inverted Microscope, and the effect on production of reactive oxygen species (ROS) was detected by flow cytometry. The mRNA expression of CYP1A and AhR was evaluated by RT-RCR. The protein expression of CYP1A1 was detected by western blot. The cell viability was significantly inhibited after HepG2 cells were exposed to dioscin 0.5-32 µmol · L(-1). Compared with the control, the LDH release rate and ROS were significantly increased. The expression of CYPlA and AhR mRNA was increased. The expression of CYP1Al protein was increased after dioscin treatment, and resveratrol, an AhR antagonist, could downregulate the expression of CYP1A1. It follows that large doses dioscin has potential hepatotoxicity. The possible mechanism may be dioscin can active aryl hydrocarbon receptor (AhR) and induce the expression of CYP1A.
Cell Survival ; drug effects ; Chemical and Drug Induced Liver Injury ; etiology ; Cytochrome P-450 CYP1A1 ; genetics ; Diosgenin ; analogs & derivatives ; toxicity ; Hep G2 Cells ; Humans ; L-Lactate Dehydrogenase ; secretion ; RNA, Messenger ; analysis ; Reactive Oxygen Species ; metabolism ; Receptors, Aryl Hydrocarbon ; genetics

To research the effect of Ginseng Radix et Rhizoma and Aconiti Lateralis Radix Praeparata compatibility on cardiac toxicity in rats by UPLC-Q-TOF/MS, and explore the endogenous markers and molecule mechanism. Different compatibility of Shenfu decoction were given to male Wistar rats at dosage of 20 g · kg(-1) for 7 days, collected the serum, and analyze the endogenous metabolites effected by Shenfu formulation by principal component analysis and partial least-squares analysis. Results showed that content of glutathione, phosphatidylcholine and citric acid decreased in mixed-decoction group, while ascorbic acid, uric acid, D-galactose, tryptophan, L-phenylalanine increased. The results showed cardiac toxicity of Aconiti Lateralis Radix Praeparata in Shenfu mixed-decoction. Shenfu co-decoction group showed a similar or weaker trend compared with control group, but most of them do not have a statistically significant. The results indicated the scientific basis of Shenfu compatibility by comparison of co-decoction group with mixed-decoction group. Shenfu compatibility can reduce cardiac toxicity induced by Aconiti Lateralis Radix Praeparata, and citric acid, glutathione, phosphatidyl choline, uric acid might be regarded as potential markers of cardiotoxicity.
Animals ; Biomarkers ; Cardiotoxicity ; Drugs, Chinese Herbal ; toxicity ; Glutathione ; blood ; Least-Squares Analysis ; Male ; Metabolomics ; methods ; Principal Component Analysis ; Rats ; Rats, Wistar

OBJECTIVETo establish the HPLC fingerprint of Rhizoma Polygoni Cuspidati (Polygonum cuspidatum).

METHODThe HPLC separation was carried with Diamonsil C18 column and eluted with a gradient from methanol and 0.1% phosphoric acid, the detection wavelength was at 230 nm and recording 70 min. The similarity of chromatograms was compared by mean of the software from Zhongnan University.

RESULTThe constituents of P. cuspidatum were well separated by HPLC, and the similarity was above 0.80.

CONCLUSIONThe method can be used for the study of fingerprints of Rhizoma Polygoni Cuspidati.

Chromatography, High Pressure Liquid ; methods ; Emodin ; analogs & derivatives ; analysis ; Fallopia japonica ; chemistry ; classification ; Glucosides ; analysis ; Plants, Medicinal ; chemistry ; classification ; Quality Control ; Reproducibility of Results ; Rhizome ; chemistry ; Stilbenes ; analysis ; Technology, Pharmaceutical ; methods

To study the effect of Siwu decoction on the function and expression of P-glycoprotein (P-gp) in Caco-2 cells. The Real-time quantitative poly-merase chain reaction (Q-PCR) was used to analyze the mRNA expression of MDR1 gene in Caco-2 cells. Flow cytometer was used to study the effect of Siwu decoction on the uptake of Rhodamine 123 in Caco-2 cells, in order to evaluate the efflux function of P-gp. Western blotting method was used to detect the effect of Siwu decoction on the P-gp protein expression of Caco-2 cells. Compared with the blank control group, after Caco-2 incubation with Siwu decoction at concentrations of 3.3, 5.0, 10.0 g x L(-1) for 24, 48, 72 h, the mRNA expression of MDR1 was up-regulated, suggesting the effect of Siwu decoction in inducing the expression of MDR1. After the administration with Siwu decoction in Caco-2 cells for 48 h, the uptake of Rhodamine 123 in Caco-2 cells decreased by respectively 16.6%, 22.1% (P < 0.05) and 45.4% (P < 0.01), indicating that the long-term administration of Siwu decoction can enhance the P-gp efflux function of Caco-2 cells. After the incubation of Caco-2 cells with Siwu decoction for 48 h, the P-gp protein expression on Caco-2 cell emebranes, demonstrating the effect of Siwu decoction in inducing the protein expression of P-gp.
ATP Binding Cassette Transporter, Sub-Family B ; genetics ; metabolism ; ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; metabolism ; Caco-2 Cells ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Up-Regulation ; drug effects

AIMThrough studying local regulatory mechanisms in pulmonary arteries (PA) and thoracic aortae (TA) under simulated microgravity (SM), to collect some data for the researches of adaptive mechanisms in pulmonary and systemic arteries and for the mechanisms accounting for orthostatic intolerance after SM.

METHODSCardiopulmonary circulatory function during 7-day 6 degrees head down bed rest (HDT) in male young volunteers was measured with a XXH-2000 pulmonary circulation and cardiac function instrument. - 30 degrees C tail suspended (TS) rats were used as the model to simulate the physiological effects of M. The PA and TA changes of vasoreactivity were respectively observed by vitro vessel rings perfusion.

RESULTSThe changes in volume of PA and pulmonary vein during a cardiac cycle and the preload in left cardiac ventricle in men increased significantly in the initial HDT. The super-regulatory phenomena appeared in both pulmonary and systemic circulation, but earlier and more obviously in pulmonary circulation than systemic circulation during 96-144 h. The dilatory reactivity in TS7 PA increased significantly, tended to decrease in TS14. The dilatory reactivity of TA in TS7 had a significant increase, had a slight increase in TS14. The contractile reactivity of PA decreased slightly in TS7 from CON, and were attenuated significantly in TS14. The contractile reactivity of TA in TS14 decreased significantly. The responsiveness to KCl, phenylephrine and sodium nitroprusside in VEC- removed PA had no differences among all groups.

CONCLUSIONThe differences in changes between pulmonary and systemic arteries under SM could be an important sign of depressed local regulatory function, which might be mainly due to dilatory function in VEC and contribute to the occurrence of orthostatic intolerance after SM.

Animals ; Aorta, Thoracic ; physiology ; Humans ; Male ; Pulmonary Artery ; physiology ; Rats ; Rats, Wistar ; Vascular Resistance ; Weightlessness ; Weightlessness Simulation ; Young Adult

OBJECTIVETo construct an eukaryotic expression plasmid containing gp120 gene of HIV-1 subtype B and obtain gp120 gene expression in HepG2 cells.

METHODSAccording to the published gp120 gene sequence in Genbank, a pair of primers was designed and synthesized. The PCR amplification product of gp120 gene was cloned into pMD-18T vector using TA cloning followed by BamHI and XhoI digestion and sequence analysis. The target gene was then subcloned into a highly efficient eukaryotic expression vector pcDNA3.1 (+). The recombinant plasmid was sequenced and identified by restrictive endonuclease digestion, and transfected into HepG2 cells via liposome. The expression of gp120 gene was analyzed by RT-PCR and Western blotting, respectively.

RESULTSRestriction endonuclease digestion and sequence analysis verified successful construction of the recombinant vector pcDNA3.1(+)/gp120. The target fragment gp120 was identical with U26942 in Genbank, and the expression of gp120 gene was detected in the lysate of the transfected HepG2 cells by RT-PCR and Western blotting.

CONCLUSIONThe eukaryotic expression plasmid for gp120 has been constructed successfully, which is capable of stable expression in HepG2 cells.

AIDS Vaccines ; biosynthesis ; genetics ; Base Sequence ; Carcinoma, Hepatocellular ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Cloning, Molecular ; Eukaryotic Cells ; metabolism ; Gene Expression ; HIV Envelope Protein gp120 ; biosynthesis ; genetics ; HIV-1 ; genetics ; Humans ; Liver Neoplasms ; genetics ; metabolism ; pathology ; Molecular Sequence Data ; Plasmids ; genetics ; Transfection ; Vaccines, DNA ; biosynthesis ; genetics

AIMTo explore the influence of hyperbaric oxygen (HBO) preconditioning on anoxic resistance and anti-weariness at high altitude.

METHODS(1) SOD, MDA, NO, NOS, BLA and BUN of 20 youths living at 3 700 m altitude for half year were tested, then they were divided into group A (n=10, received HBO pretreatment twice) and group B (n=10, received HBO pretreatment 5 times) randomly. They were asked to pedal the EMG-bicycle-ergometer at the second and eighth day, and then the same items were tested. (2) 29 youth who would go to Astronomical Spot (5200 m) were randomly divided into group HBO (n=11, received HBO pretreatment once per day for 2 days at Yecheng (1400 m)) and comparison group (n=10). When they reached I Astronomical Spot, thematic biochemical index were investigated. (3) When 20 youth reached Thirty Milepost Barracks (3700 m) at the second day in their way to Immortal Gulf (5380 m) from Yecheng were randomly divided into group HBO (n=10, received HBO pretreatment once per day for 3 days) and comparison group (n=10). When they reached Immortal Gulf, the thematic biochemical index were investigated.

RESULTS(1) SOD, NO, NOS were increased and BLA, BUN, MDA were decrease in group A compared with that in group B until the eighth day, there was significant difference (P < 0.01). (2) SOD, NO, NOS were increased and BLA, BUN, MDA were decrease in group HBO compared with that in comparison that in group, there was significant difference between groups (P < 0.01, or P < 0.05).

CONCLUSIONHBO could enhance the activity of anti-oxidase and the cleared ability of lactic acid, and the effect of anti-weariness could last for 8 days.

Altitude ; Altitude Sickness ; prevention & control ; Fatigue ; prevention & control ; Humans ; Hyperbaric Oxygenation ; methods ; Hypoxia ; physiopathology ; Ischemic Preconditioning ; methods ; Male ; Oxidative Stress ; drug effects ; physiology ; Superoxide Dismutase ; metabolism ; Young Adult