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

Gushukang is a compound Chinese herbal preparation. Pharmacological studies indicated that Gushukang can increase bone density, inhibit bone resorption, promote bone formation, and restore bone microstructure, as well as improve bone biomechanical parameters and promote healing of bone fracture. Clinical observations demonstrated that it has favorable efficacy in preventing and treating osteoporosis.
Bone Density ; drug effects ; Bone Resorption ; prevention & control ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Osteogenesis ; drug effects ; Osteoporosis ; drug therapy ; physiopathology ; Phytotherapy

To explore the prevention effect of the joint combination of Yindanxinnaotong soft capsule (YDXNT) and exercise (swimming) on atherosclerotic rats. The method of 3 x 3 factorial design, including two factors (YDXNT and swimming) and three levels (0, 1, 2 g x kg(-1) YDXNT; 0, 0.5, 1 h swimming), was mainly adopted. The atherosclerotic rat model was established by ligating their left common carotid arteries and feeding high-fat diet. After 8 weeks, blood samples were collected from their thoracic aorta to determine blood viscosity, plasma viscosity, fibrinogen (FIB), nitric oxide (NO), 6-keto-PGF(1alpha) endothelin (ET) and thromboxane B2 (TXB2). The tissues of left common carotid arteries of the rats were collected to detect the positive expression of SM22alpha and determine the semi-quantitation through the immunohistochemical staining. The result showed that the combination of YDXNT and swimming can significantly decrease the plasma viscosity (F = 3.241, P = 0.017), the high and low shear blood viscosity (F = 6.444, P = 0.001; F = 3.002, P = 0.024) and FIB (F = 4.046, P = 0.005). The increased NO and 6-keto-PGF(1alpha) and the decreased ET and TXB2 indicated a significant interaction (P < 0.05). The swimming showed an obvious main effect in the expression of up-regulated protein SM22alpha (F = 8.088, P = 0.001). The study suggested that the combined administration of YDXNT and swimming could improve the hemorheological parameters of atherosclerotic rats, protect the vascular endothelium, inhibit the vascular remodeling in atherosclerosis and positively prevent the atherosclerosis.
Animals ; Atherosclerosis ; drug therapy ; genetics ; metabolism ; prevention & control ; Blood Viscosity ; drug effects ; Combined Modality Therapy ; Drugs, Chinese Herbal ; administration & dosage ; Exercise Therapy ; Humans ; Male ; Microfilament Proteins ; genetics ; metabolism ; Muscle Proteins ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Swimming

AIMTo investigate the inhibitory effect of egg white lysozyme (LZM) on ceftazidime (CFT)-induced release of endotoxin from Pseudomonas aeruginosa.

METHODSP. aeruginosa PAO1 was inoculated in nutrition broth or diluted rabbit blood free of antibiotics in the presence or absence of LZM and incubated at 37 degrees C on a water bath shaker. beta-Lactam antibiotic, CFT, was added to cultures at 3.5 h (nutrition broth culture) or 5 h (diluted rabbit blood culture) after inoculation. After 3 h of CFT treatment, the supernatants from different bacterial cultures were prepared by centrifuge and the concentrations of endotoxin in the supernatants were measured. The bacterial supernatants were also added to a murine macrophage cell line RAW 264.7 or intravenously injected into carrageenin-sensitized mice. Tumor necrosis factor-alpha (TNF alpha) and nitric oxide (NO) concentrations in RAW 264.7 supernatants or in mouse sera were tested.

RESULTSCFT treatment alone obviously inhibited the growth of P. aeruginosa PAO1 accompanied by strong and rapid bacteriolysis and released relatively high concentration of endotoxin from bacteria both in nutrition broth and in diluted rabbit blood cultures. The bacterial supernatant from CFT treatment alone yielded high concentrations of TNF alpha both in RAW 264.7 cells and in mice and high level of NO in RAW 264.7 cells. Treatment with the combination of LZM and CFT evidently blocked the lysis of bacteria and reduced the release of endotoxin without decreasing bactericidal activity of CFT. TNF alpha and NO productivity of the supernatants prepared from the LZM/CFT combinative treated bacterial cultures were significantly decreased both in RAW 264.7 cells and in mice indicating that the inflammatory activity was reduced.

CONCLUSIONLZM can effectively prevent CFT-induced bacteriolysis, endotoxin release and subsequent proinflammatory factor production but without decreasing bactericidal activity of CFT, resulting in the disassociation of bactericidal activity and bacteriolysis. Thus, LZM might be important for preventing endotoxemia in Gram-negative sepsis with the treatment of antibiotics.

Animals ; Bacteriolysis ; drug effects ; Ceftazidime ; pharmacology ; Egg White ; analysis ; Endotoxins ; metabolism ; Mice ; Muramidase ; isolation & purification ; pharmacology ; Nitric Oxide ; metabolism ; Pseudomonas aeruginosa ; metabolism ; physiology ; Rabbits ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo compare therapeutic effects of traditional moxibustion and moxibustion on temperature-sensitive points for lumbar disc herniation.

METHODSOne hundred and twenty cases were randomly divided into a temperature-sensitive point group and a traditional moxibustion group, 60 cases in each group. The temperature-sensitive point group was treated with moxibustion on the temperature-sensitive points in the temperature-sensitive high incidence area such as waist and lower limbs, once each day; the traditional moxibustion group was treated with warming moxibustion at Jiaji (EX-B 2), Ciliao (BL 32), Zhibian (BL 54), Huantiao (GB 30), Weizhong (BL 40), Yanglingquan (GB 34), Kunlun (BI. 60), once each day, 7 days constituting one course. The therapeutic effect and the recurrence rate were observed after one course treatment and six months later.

RESULTSAfter treatment for one course, the cured-markedly effective rate was 65.00% in the temperature-sensitive point group, superior to 50.0% in the traditional moxibustion group, with a significant difference between the two groups (P < 0.05); all effective cases were followed-up for six months, the cured-markedly effective rate and the recurrence rate were 62.3% and 26.4% in the temperature-sensitive point group, and 34.2% and 46.3% in the traditional moxibustion group, the therapeutic effect of the temperature-sensitive point group being superior to the traditional moxibustion group, and the recurrence rate was lower than the traditional moxibustion group (both P < 0.05).

CONCLUSIONMoxibustion on temperature-sensitive points is a effective therapy for lumbar disc herniation, with stable therapeutic effect and low recurrence rate.

Acupuncture Points ; Adult ; Female ; Follow-Up Studies ; Humans ; Intervertebral Disc Displacement ; therapy ; Lumbar Vertebrae ; Male ; Middle Aged ; Moxibustion ; methods ; Temperature ; Treatment Outcome ; Young Adult

OBJECTIVETo establish a LC-MS/MS method for determining the concentration of tanshinone IIA, salvianolic acid B and paeoniflorin of refined coronary cataplasm in rabbit plasma, in order to determine the concentration of the three main ingredients in blood after transdermal administration and calculate their pharmacokinetic parameters.

METHODRabbits were given refined coronary cataplasm on the basis of 15 g x kg(-1) by transdermal administration to detect the plasma concentration of the three main ingredients using LC-MS/MS. Winnonlin software was used to calculate their major pharmacokinetic parameters.

RESULTTanshinone IIA, salvianolic acid B and paeoniflorin showed good linearity (r>0.999) at 1-100, 50-1 000, 10-1 000 microg x L(-1) respectively in plasma, with average recovery rate of 96.57%, 91.90%, 95.93%, respectively. The RSD within day were less than 15%. After transdermal administration of refined coronary cataplasm in rabbits, the main pharmacokinetic parameters of tanshinone IIA, salvianolic acid B or paeoniflorin were as follows: Cmax (20.85 +/- 12.68), (636.25 +/- 386.91), (787.80 +/- 395.64) microg x L(-1); Tmax (0.49 +/- 0.28), (0.44 +/- 0.27), (0.46 +/- 0.30) h.

CONCLUSIONThe LC-MS/MS method is highly selective and sensitive to determine the concentration of samples in rabbit plasma. The pharmacokinetic characteristics of tanshinone IIA, salvianolic acid B and paeoniflorin are suitable to assess the percutaneous absorption of refined coronary cataplasm.

Animals ; Benzoates ; administration & dosage ; pharmacokinetics ; Benzofurans ; administration & dosage ; pharmacokinetics ; Bridged-Ring Compounds ; administration & dosage ; pharmacokinetics ; Chromatography, Liquid ; methods ; Coronary Disease ; drug therapy ; Diterpenes, Abietane ; administration & dosage ; pharmacokinetics ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Glucosides ; administration & dosage ; pharmacokinetics ; Humans ; Male ; Monoterpenes ; Rabbits ; Tandem Mass Spectrometry ; methods

OBJECTIVETo observe the antioxidant effects of water extract of Carthamus tinctorius on ox-LDL induced injury in rat cardiac microvascular endothelial cell and detecting oxygen derived free radicals (OFR) to explore the antioxidant mechanisms.

METHODBy using the third generation of rat cardiac microvascular endothelial cells (rCMEC), the protection of water extract of C. tinctorius was investigated after ox-LDL (100 mg x L(-1) induced damage. The supernatant was collected for detecting lactate dehydrogenase (LDH), malondialdehyde (MDA), superoxide dismutase (SOD), xanthine oxidase (XOD), glutathione peroxidase (GSH-Px), nitric oxide (NO), nitric oxide synthase (NOS) activity, and cell suspension was collected for detecting reactive oxygen species (ROS) by electron spin resonance (ESR).

RESULTWater extract of C. tinctorius increased the rCMEC survival rate, reduced LDH, MDA and XOD levels, and improved SOD, GSH-Px and NOS activity, while in the cell suspension ROS signal decreased significantly.

CONCLUSIONWater extract of C. tinctorius has antioxidation. The mechanisms are likely related with scavenging of free radicals, enhancing its clearance, enhancing endogenous antioxidant activity.

Animals ; Carthamus tinctorius ; chemistry ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Electron Spin Resonance Spectroscopy ; methods ; Endothelial Cells ; drug effects ; metabolism ; Enzyme Activation ; drug effects ; Glutathione Peroxidase ; metabolism ; L-Lactate Dehydrogenase ; metabolism ; Lipoproteins, LDL ; pharmacology ; Microvessels ; cytology ; Nitric Oxide Synthase ; metabolism ; Rats ; Rats, Wistar ; Reactive Oxygen Species ; metabolism ; Superoxide Dismutase ; metabolism ; Water ; chemistry ; Xanthine Oxidase ; metabolism

OBJECTIVETo observe the protective effects of catechin morphon (GCG and EGCG) on hypoxia-reoxygenation induced injury in myocardial cells and to explore the mechanisms.

METHODIn cultured neonatal rat cardiomyocytes, we investigated the preconditioning protection by GCG and EGCG on the spontaneous beating, the survival rate, the release of LDH, MDA, SOD, GSH-Px and the ATP enzyme activity of cardiomyocyte cellular membrane in cultured rat cardiomyocytes treated during the reoxygenation 1h following hypoxia 3 h. The blocking agent of protien kinase C staurosporine (10 nmol x L(-1)) or the deactivator of Gi/o protein pertussis toxin (PTX, 200 microg x mL(-1)) were added before the catechin treatment.

RESULTSPreconditioning by GCG and EGCG increased the spontaneous beating and the survival rate, and decreased the release of LDH and MDA with the rise of SOD and ATP enzyme activity. Inhibition of PKC by staurosporine and Gi/o protein by PTX abolished the protection by catechin with the reduction of the beating, survival rate and activity of SOD, and the increase of the release of LDH and MDA. The results indicated that the activation of signal transduction pathway from PKC and Gi/o protein seemed to be involved in the cardioprotection of preconditioning by GCG and EGCG.

CONCLUSIONThe protection by GCG and EGCG on hypoxia-reoxygenation injury in cultured neonatal rat cardiomyocytes is found, which is related with scavenging of free radicals, and PKC Gi/o signal transduction pathway.

Animals ; Cardiotonic Agents ; chemistry ; pharmacology ; Catechin ; analogs & derivatives ; chemistry ; pharmacology ; Female ; Free Radicals ; metabolism ; Hypoxia ; complications ; Myocytes, Cardiac ; drug effects ; metabolism ; pathology ; Oxygen ; metabolism ; Protein Kinase C ; metabolism ; Proteins ; metabolism ; Rats ; Signal Transduction ; drug effects ; Survival Rate

OBJECTIVETo study on the chemical stability of Salvia miltirrhiza hairy root.

METHODThe rolA gene was detected by PCR in DNA and the chemical contituent variances were detected by HPLC.

RESULTThe rolA gene was found in all the 10 batches of the culfured hairy root. The similarities of the chromatographic fingerprints of the 10 batches are higher than 0. 95.

CONCLUSIONThere are no significant differences of the chemical constituents in 10 hairy root samples.

Chromatography, High Pressure Liquid ; DNA, Plant ; genetics ; Drugs, Chinese Herbal ; chemistry ; Genes, Plant ; Plant Roots ; chemistry ; genetics ; Polymerase Chain Reaction ; Reproducibility of Results ; Salvia miltiorrhiza ; chemistry ; genetics ; Sensitivity and Specificity

OBJECTIVETo compare the difference in clinical efficacy on lumber disc herniation (LDH) treated with Yaoyangguan (GV 3) between mild moxibustion under thermosensitive condition and that under non-thermo-sensitive condition.

METHODSFifty-seven LDH patients were selected as the study objects. Mild moxibustion at Yaoyangguan (GV 3) was applied for 45 min each time. Additionally, the conventional acupuncture was given, once a day, for 20 days. At the end of treatment, two groups were classified. A thermosensitive acupoint group (thermosensitive group) was composed of the cases with thermo-sensitization at Yaoyangguan (GV 3) and presenting for > or =4 times in the entire treatment. A non-thermosensitive acupoint group (tranquilization group) was composed of the cases without thermo-sensitization or the frequency of thermo-sensitization <4 times in the entire treatment. The modified Japanese orthopedics association scoring system (M-JOA) was adopted to observe the cases before and after treatment and 6 months after treatment in the two groups. The efficacy was compared between the two groups.

RESULTSAfter treatment and in 6 months after treatment, the score of M-JOA was apparently reduced as compared with that before treatment in the two groups (all P<0.05). The score reducing in the thermosensitization group was more obvious than that in the tranquilization group (both P<0.01). After treatment and in 6 months after treatment, the curative rate and remarkably effective rate were 89.7% (26/29) and 79.3% (23/29), which was better than 71.4% (20/28) and 60.7% (17/28, both P<0.05) in the tranquilization group separately.

CONCLUSIONMild moxibustion at acupoint under thermosensitive condition achieves the better short-term and long-term effects as compared with that under non-thermosensitive condition.

Acupuncture Points ; Adult ; Female ; Humans ; Intervertebral Disc Displacement ; therapy ; Male ; Middle Aged ; Moxibustion ; Treatment Outcome