[Prediction and analysis of Q-markers of Elephantopus scaber based on its UPLC fingerprint, content determination of components, and in vitro a nti-tumor activity].

This study aimed to provide scientific evidence for predicting quality markers(Q-markers) of Elephantopus scaber by establishing UPLC fingerprint of E. scaber from different geographical origins and determining the content of 13 major components, as well as conducting in vitro anti-cancer activity investigation of the main components. The chromatographic column used was Waters CORTECS UPLC C_(18)(2.1 mm×150 mm, 1.6 µm), and the mobile phase consisted of acetonitrile and 0.1% formic acid solution(gradient elution). The column temperature was set at 30 ℃, and the flow rate was 0.2 mL·min~(-1). The injection volume was 1 µL, and the detection wavelength was 240 nm. The UPLC fingerprint of E. scaber was fitted using the Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine(2012 edition) to determine common peaks, evaluate similarity, identify and determine the content of major components. The CCK-8 assay was used to explore the inhibitory effect of the main components on the proliferation of lung cancer cells. The results showed that in the established UPLC fingerprint of E. scaber, 35 common peaks were identified. Thirteen major components, including neochlorogenic acid(peak 1), chlorogenic acid(peak 2), cryptochlorogenic acid(peak 3), caffeic acid(peak 4), schaftoside(peak 6), galuteolin(peak 9), isochlorogenic acid B(peak 10), isochlorogenic acid A(peak 12), isochlorogenic acid C(peak 18), deoxyelephantopin(peak 28), isodeoxyelephantopin(peak 29), isoscabertopin(peak 31), and scabertopin(peak 32) were identified and quantified, and a quantitative analysis method was established. The results of the in vitro anti-cancer activity study showed that deoxyelephantopin, isodeoxyelephantopin, isoscabertopin, and scabertopin in E. scaber exhibited inhibition rates of lung cancer cell proliferation exceeding 80% at a concentration of 10 µmol·L~(-1), higher than the positive drug paclitaxel. These results indicate that the fingerprint of E. scaber is highly characteristic, and the quantitative analysis method is accurate and stable, providing references for the research on quality standards of E. scaber. Four sesquiterpene lactones in E. scaber show significant anti-cancer activity and can serve as Q-markers for E. scaber.

[Progranulin attenuated asthma inflammation by inhibiting the expression of IL-6].

Objective: To investigate the role and mechanism of progranulin (PGRN) in asthma. Methods: Control group and model group were set up in wild and IL-6 knockout (IL-6 ko) mice, respectively. For asthma model, mice were intraperitoneally sensitized with 100 µg OVA on days 0 and 7, followed by aerosol challenges with 5% OVA for 30 min per day from day 14 to 21, and mice were sacrificed 24 h after the last challenge. The mice in control group were treated in the same way with PBS. Bronchoalveolar lavage fluid (BALF) was collected for leukocytes count and differential count. The pathological changes of lung tissues were observed by H&E staining. The cytokines in lung homogenate, serum and BALF were detected by Q-PCR and ELISA. The in vitro model of asthma was induced by stimulating A549 or BEAS-2B cells with IL-13. Each group was replicated in three wells and four groups were designed: PBS group, IL-13 treatment group, IL-13 + rhPGRN treatment group, inhibitors of p38 phosphorylation (SB203508) treatment group. The cells or supernatant were collected after 0~48 h. PGRN and IL-6 levels were determined by Q-PCR and ELISA, the level of p38 phosphorylation was tested by Western blot (WB). Results: Compared with control group, PGRN levels were decreased in lung homogenate and BALF (P＜0.05), and PGRN presented a downtrend in serum, however, the level of IL-6 in BALF was increased in asthma mice (P＜0.01). In IL-6 ko asthma mice, compared with the wild asthma mice, leukocytes, especially neutrophils in BALF were decreased (P＜0.05), but PGRN was increased (P＜0.05), lung pathological damage was significantly alleviated. In vitro experiments, compared with PBS group, PGRN level was decreased (P＜0.05), IL-6 level was increased (P＜0.01), phosphorylation of p38 was activated in IL-13 treatment group. Compared with IL-13 treatment group, in IL-13 + PGRN treatment group, IL-6 level was decreased (P＜0.05); phosphorylation of p38 was inhibited (P＜0.05); and the production of IL-6 (P＜0.05) was decreased after treatment with inhibitor of p38 phosphorylation. Conclusion: PGRN inhibited the production of IL-6 by suppressing the p38 phosphorylation to alleviate asthmatic airway inflammation.

[Comparison of trophic status analysis of the Daning River within the Three Gorges Reservoir before and after experimental impoundment].

We evaluated 4-year data set to assess the trophic state and limiting factors of the Three Gorges Reservoir (TGR) during the experimental impounding period (September 2005 to September 2007) and the normal operating period (September 2008 to September 2010). The results indicated that there had been appeared new characteristics in spatial and temporal distribution of trophic state indices after impoundment. The trophic state indices (TSI(TP)) showed increased trend after the TGR impoundment during the study area, but TSI(TN) and TSI(SD) had no significant changes after the TGR impoundment. The values of TSI(CHL) showed increased trend after the TGR impoundment in S1, and the values of TSI(CHL) did not show obvious changes in S2, S3 and S4 after the TGR impoundment. The values of TSI(TN), TSI(TP) and TSI(SD) show similar spatial variances with the highest value in S4, followed in a descending order by S3, S2 and S1. TSI(CHL) in the S2 and S3 were higher than that in S1 and S4. According to the characteristics of water level, the operational period of the TGR classified into following four stages: stage I (pre-November-April), stage II (May-July), stage III (July-September) and stage IV (September-November). The values of TSI(TN) and TSI(TP) in the Daning River and the TGR mainstream showed similar seasonal variances with the highest value in the stage II and III, followed in a descending order by stage I and IV. The values of TSI(CHL) varied substantially among the four stages, with the highest value in stage III, followed by stage II, IV and I. The trophic state indices differences were getting smaller between the four stages after the TGR impoundment. Using Carlson's two-dimensional approach, deviations of the TSI(S) indicated that factors other than phosphorous and nitrogen limited algal growth and that nonalgal particles affected light attenuation. These findings were further supported by the significant correlation among the values of TSI and hydrological factors.

[Exploration of relationships between phytoplankton and related environmental factors in the Daning River during sensitive period of algal blooms].

To elucidate relationships between phytoplankton and related environmental factors, phytoplankton and environmental parameters of water quality in the Daning River were conducted using multivariate statistical analysis. Most of the investigated physico chemical parameters [water temperature, total nitrogen, nitrate-N (NO3(-) -N), pH and dissolved oxygen concentration (DO)] in Daning River are significantly different among those sampling sites (ANOVA, p < 0.05). The results indicate that the Daning River has a comparatively high spatial and temporal heterogeneity in the phytoplankton community composition (ANOVA, p < 0.05). The maximum cell density was recorded in Dachang and the minimum density was found at Wuxia-kou sites. Multi-algal species blooms were observed in the same time and place with the maximum density appeared. Correlation analysis was performed to illuminate the relationships between algae density and environmental variables, TN (r = - 0.789, p < 0.05), NO3(-) -N (r = - 0.825, p < 0.05) and NO2(-) -N (r = -0.803, p < 0.05) were negatively correlated with algae density. The results indicate that nitrogen soluble nutrients are key regulatory factors for phytoplankton abundance. The results of redundancy analysis (RDA) indicate that main soluble nutrients (TN, TP, NO3(-)-N and NO2(-)-N), suspended substance (SS) and transparency contributed significantly to phytoplankton community composition. Phytoplankton growth could enhance the pH value. The environmental characterizations and sample sites adjacency contributed significantly to phytoplankton community composition.