OBJECTIVE:To establish an HPLC method for the determination of the contents of Decussatin,Bellidifodin,Methylswertianin and Swerchirin in Swertia macrosperma C.B.Clark.METHODS:The determination was performed on VP-ODS C18(150 mm? 4.60 mm,5 ? m),with the mobile phase consisting of methanol-water-phosphoric acid(70∶ 30∶ 0.4),the detection wavelength set at 254 nm,the flow rate at 1.0 mL? min-1 and the column temperature at 20℃.RESULTS:The calibration curves of Decussating,Bellidifodin,Methylswertianin and Swerchirin were in good linearity in the range of 0.172～ 0.86 ? g,0.224～ 1.344 ? g,0.144～ 0.72 ? g and 0.154～ 0.77? g,respectively.The average recoveries were 98.18%(RSD=1.19%),99.32%(1.52%),98.44%(1.23%)and 97.68%(1.57%),respectively.CONCLUSION:The method is rapid and precise,with accurate and reliable results.

Objective To explore the alterations of resting-state neural networks in females with migraine at attack stage in low-and high-frequency ranges compared to healthy controls. Methods Resting-state magnetoencephalography data from 20 females with migraine ( migraine group ) and 20 age-and gender-matched healthy controls ( control group ) were collected to compute the neural network parameters through graph theory analysis.Independent samples t test was used to compare the difference of the network parameters between the two groups.The Pearson coefficient was used to assess the relationship between the abnormal network parameters and clinical manifestations of migraine.Results Compared with control group, females in migraine group at attack stage showed increased functional connectivity in the 0.1 -1 Hz, 4 -8 Hz, 12 -30 Hz, 30 -80 Hz band; increased shortest path length in the 80-250 Hz band; increased clustering coefficient and degree in the 0.1 -1 Hz band;decreased clustering coefficient and degree in the 4 -8 Hz, 12 -30 Hz band ( P<0.05 -0.01 ) .The functional connectivity of females with migraine at attack stage was positively correlated with number of years with migraine in the 12-30 Hz ( r=0.56, P=0.01 ) .Conclusion These results indicate that inherent neural networks are significantly impaired in low-and high-frequency ranges in females with migraine at attack stage and may contribute to the clinical manifestations of this disorder.

Objective To investigate the expressions of receptor for advanced glycation endproducts (RAGE) and nuclear factor κB(NF-κB) in brain hippocampus of rat with chronic fluorosis,and to reveal the mechanism of brain damage resulted from chronic fluorosis.Methods Sixty clean grade SD rats were randomly divided to three groups(20 rats in each group,10 female and 10 male) fed with different contents of fluoride,control group with normal tap-water(＜ 0.5 mg/L fluoride),small dosage of fluoride exposure group(10 mg/L fluoride in tap-water) and large dosage of fluoride exposure group(50 mg/L fluoride) for six months.Then the rats were killed by femoral artery bleeding and hippocampus was removed.Protein and mRNA levels of RAGE and NF-κB in the hippocampus were determined by Western blotting and quantitative real time PCR,respectively.Results As compared to the control groups[(100.00 ± 2.60)％,(100.00 ± 7.80)％],the expressions of RAGE and NF-κB at protein level in the hippocampus were significantly increased in the small dosage of fluoride exposure groups [(205.00 ± 15.30)％,(156.00 ± 12.20)％] and the large dosage of fluoride exposure groups[(232.00 ± 10.90)％,(162.00 ± 9.80)％,all P ＜ 0.05]; for the mRNA level of RAGE and NF-κB,the expressions were higher in the small dosage of fluoride exposure groups(1.27 ± 0.09,0.83 ± 0.15) and the large dosage of fluoride exposure groups (2.60 ± 0.19,1.27 ± 0.19) than those of the control groups(0.66 ± 0.18,0.32 ± 0.08,all P＜ 0.05).Conclusions The increased expressions of RAGE and NF-κB in the hippocampus of rat brain are caused by chronic fluorosis,and these changes may be associated with the mechanism of nerve injury.

Objective Aim of the study is to investigate the expression of syndecan-4 and nuclear factor κB(NF-κB) in the kidney of rat with chronic fluorosis,and to reveal the mechanism of kidney damage resulted from the toxicity of excessive amount of fluoride.Methods According to body mass and sex,sixty SD rats were randomly divided to three groups according to body mass and fed with different contents of fluoride:control group with normal tap-water(＜ 0.5 mg/L fluoride),small dosage of fluoride exposure group (adding 10 mg/L fluoride in tap-water) and large dosage of fluoride exposure group (50 mg/L fluoride) for six months.The protein level of syndecan-4 and NF-κB in the kidney was detected by Western blotting and syndecan-4 mRNA level by quantitative real time PCR.Results As compared to the control group[(100.0 + 8.1)％],the expression of syndecan-4 at protein level in the kidney of rat was significantly increased in the small dosage of fluoride exposure group [(198.5 + 5.6)％,P ＜ 0.05] and large dosage of fluoride exposure group [(209.2 + 13.0)％,P ＜ 0.05]; the protein levels of NF-κB in the small dosage of fluoride exposure group[(284.4 + 11.1)％,P ＜ 0.05] and in the large dosage of fluoride exposure group[(343.2 + 2.9)％,P ＜ 0.05] were significantly increased than that of the control group[(100.0 ± 10.7)％].The mRNA levels of syndecan-4 in the kidney in the small dosage of fluoride exposure group and large dosage of fluoride exposure group(0.431 + 0.058 and 0.453 ± 0.065,both P ＜ 0.05,respectively) were significantly increased than that of the control(0.128 + 0.026).Conclusions The increased expression of NF-κB in the kidney is induced by increased expression of syndecan-4,which may be involved in kidney damage of chronic fluorosis.

Objective: To seek the optimum technology of drying and to solve the problem of powder sticking clump. Methods : On the basis of major drying condition, such as duration of drying, temperature of drying, evaluating different drying technology; On the basis of the quantity of the powder sticking on the wall and the hygroscopicity of one, investigating different excipients how to affect the spray drying. Seeking the optimum kinds and quantity of excipients. Results : The best properties of the granules are from the spray drying process. ?-cyclodextrin and lactose have the effects on solving the powder sticking on the wall. Conclusion : The spray drying process is the optimum one. Adding 3～5 percent of ?-cyclodextrin has an effect on improving the phenomenon of powder sticking on the wall in process of the spray drying.

AIM: To study the antioxidative effect of three kinds of xanthone:methylswertianin(MET), bellidifodin(BEL) and decussatin (DEC) extracted from Swertia macrosperma C.B. clark. METHODS: Hydroxy radical was generated from H_2O_2 and VC-Fe~ 2+ system; MDA contents in liver homogenate and mitochondria were measured by thiobabituric acid assay; the hemolysis of RBC and the swelling extent of mitochondria were detected by spectrophotometric methods. RESULTS: Xanthones could inhibit the generation of MDA, the hemolysis of RBC, and the swelling of mitochondria. The inhibitory effect of BEL was stronger than that of MET and DEC. CONCLUSION: Xanthones possess a good antioxidant activity.

Animals ; Brain ; drug effects ; metabolism ; Calmodulin ; metabolism ; Female ; Lead ; toxicity ; Male ; Maternal Exposure ; Pregnancy ; Protein Kinase C ; metabolism ; Rats ; Rats, Wistar

Ischemic heart disease (IHD), which has been considered to be exclusively caused by stenosis or occlusion of coronary artery, is a significant cause of morbidity and mortality worldwide. Mitochondrial dysfunction is the main pathological basis of ischemic heart disease and reperfusion injury, and moderate mitochondrial autophagy can selectively remove damage proteins and organelles to maintain intracellular homeostasis, so mitochondrial autophagy is important for maintaining the homeostasis of cardiomyocytes. Natural drugs from plants are widely used in ischemic heart disease. In recent years, more and more natural drugs have been proven to alleviate myocardial cell damage after ischemia/reperfusion through mitochondrial autophagy. This paper reviews the research progress of natural drugs from plants medicines regulating mitochondrial autophagy in the treatment of ischemia heart disease.

Objective The aim of the study was to investigate the expression of P-glycoprotein (P-gp)and nuclear factor κB (NF-κB) in the cerebral cortex of rat with chronic fluorosis,and to reveal the mechanisms of damaged nervous system resulted from the toxicity of fluoride.Methods Sixty SD rats were randomly divided into three groups.The rats in each group were given drinking water containing different levels of fluoride:control group less than 0.5 mg/L,small amount of fluoride exposure group 10.0 mg/L and large amount of fluoride exposure group 50.0 mg/L.The animals were examined at the sixth month after initiating the experiment.Protein levels of P-gp and NF-κB in brain tissues were detected by immunocytochemistry and Western blotting,and the P-gp protein and mRNA level by quantitative real time PCR method.Results As compared to the control group(28.21 ±6.13),the numbers of positive staining cells by P-gp antibody in the cortex of rat brains were significantly increased in both the small and the large amount of fluoride exposure groups[(48.46 ± 8.00),(53.72 ± 9.15),respectively,all P ＜ 0.05] ; the protein levels in the control group(100.00 ± 3.86)％ detected by Western blotting were significantly increased in the cortex of rat brains treated with fluoride in both the small and the large amount of fluoride exposure groups[(189.47 ± 3.14)％,(191.36 ± 11.09)％,respectively,all P ＜ 0.05].The significantly increased expression of NF-κB at the protein level was observed in the cortex of rat brains of the small and the large amount of fluoride exposure groups[(365.97 ± 6.04)％ and (417.15 ± 10.89)％,respectively] as compared with the control group[(100.00 ± 10.07)％,all P ＜ 0.05].The mRMA level of P-gp in the cortex of rat brains of the small and the large amount of fluoride exposure groups(2396 ± 427,3479 ± 371,respectively) were higher than that of the control group(260 ± 106,all P ＜ 0.05).Conclusion The increased expressions of P-gp and NF-κB in the cortex of rat brains are induced by chronic fluorosis,which might be connected with the mechanism of brain damages.

Objective To investigate the changes of reactive oxygen species(ROS) level and mitochondria fission-fusion-balance in cortical neurons of rats with chronic fluorosis and reveal the correlation between these two factors. Methods One hundred and twenty rats were randomly divided into 3 groups(control group, low-dose fluorosis group, high-dose fluorosis group) and 40 rats were in each group according to body weight and the experiments were carried out for 3 months or 6 months. The rats were fed with different concentrations of fluoride (NaF) to establish fluorosis models. Controls were fed with tap water( < 0.5 mg/L), experimental animals in low- or high-dose group were fed with water containing NaF 10.0,50.0 mg/L, respectively. The level of ROS and the morphology in mitochondria fission-fusion balance in neurons of the cortex of rat brains prepared with cortical frozen sections were detected with ROS fluorescent probe and MitoTracker RED probe, respectively. Results Significant differences of the level of ROS and the numbers of abnormal mitochondria in morphology in the cortical neurons were found between 3 groups at the experiment period of 3 month and 6 month(F= 3.07,3.06,3.05,3.07, all P < 0.05). As compared with control group(10.43 ± 5.98,4.12 ± 3.86) at the experiment period of 3 month, the level of ROS and the numbers of abnormal mitochondria in morphology in the cortical neurons were obviously increased in high-dose fluorosis group(25.48 ± 6.09,20.47 ± 6.09, all P < 0.05), whereas no significant changes were found in low-dose fluorosis group(11.67 ± 3.49,6.68 ± 3.48, all P> 0.05). Furthermore, the increases in both ROS level and abnormal numbers of mitochondria were significant observed in the cortical neurons of low-dose fluorosis group (63.02 ± 8.15, 49.33 ± 8.61) and high-dose fluorosis group(65.60 ± 7.40,53.10 ± 6.95) as compared with the control group (25.26 ± 6.41,20.26 ± 6.41) at the experimental period of 6 month (all P < 0.05). The abnormal numbers of mitochondria correlated with ROS level(r = 0.93,0.81, all P < 0.05). Conclusions Taking excessive amount of fluoride results in high level of oxidative stress and impaired the balance of mitochondrial fission-fusion,which is dependent on the feeding times and doses of fluoride. The mechanism of the mitochondrial abnormalities might be associated with the high level of oxidative stress induced by chronic fluorosis.