Ethyl Lithospermate Reduces Lipopolysaccharide-Induced Inflammation through Inhibiting NF-κB and STAT3 Pathways in RAW 264.7 Cells and Zebrafish.

OBJECTIVE: To explore the anti-inflammatory effects of ethyl lithospermate in lipopolysaccharide (LPS)-stimulated RAW 264.7 murine-derived macrophages and zebrafish, and its underlying mechanisms. METHODS: 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazoliumbromide (MTT) assays were performed to investigate the toxicity of ethyl lithospermate at different concentrations (12.5-100 µ mol/L) in RAW 264.7 cells. The cells were stimulated with LPS (100 ng/mL) for 12 h to establish an inflammation model in vitro, the production of pro-inflammatory cytokines interleukin (IL)-6 and tumor necrosis factor α (TNF-α) were assessed by enzyme linked immunosorbent assay (ELISA). Western blot was used to ascertain the protein expressions of signal transducer and activator of transcription 3 (STAT3), nuclear factor kappa B (NF-κB) p65, phospho-STAT3 (p-STAT3, Tyr705), inhibitor of NF-κB (IκB) α, and phospho-I κB α (p-IκB α, Ser32), and confocal imaging was used to identify the nuclear translocation of NF-κB p65 and p-STAT3 (Tyr705). Additionally, the yolk sacs of zebrafish (3 days post fertilization) were injected with 2 nL LPS (0.5 mg/mL) to induce an inflammation model in vivo. Survival analysis, hematoxylin-eosin (HE) staining, observation of neutrophil migration, and quantitative real-time polymerase chain reaction (qRT-PCR) were used to further study the anti-inflammatory effects of ethyl lithospermate and its probable mechanisms in vivo. RESULTS: The non-toxic concentrations of ethyl lithospermate have been found to range from 12.5 to 100 µ mol/L. Ethyl lithospermate inhibited the release of IL-6 and TNF-α(P<0.05 or P<0.01), decreased IκBα degradation and phosphorylation (P<0.05) as well as the nuclear translocation of NF-κB p65 and p-STAT3 (Tyr705) in LPS-induced RAW 264.7 cells (P<0.01). Ethyl lithospermate also decreased inflammatory cells infiltration and neutrophil migration while increasing the survival rate of LPS-stimulated zebrafish (P<0.05 or P<0.01). In addition, ethyl lithospermate also inhibited the mRNA expression levels of of IL-6, TNF-α, IκBα, STAT3, and NF-κB in LPS-stimulated zebrafish (P<0.01). CONCLUSION: Ethyl lithospermate exerts anti-Inflammatory effected by inhibiting the NF-κB and STAT3 signal pathways in RAW 264.7 macrophages and zebrafish.

Isoquinoline alkaloids from Corydalis edulis Maxim. Exhibiting insulinotropic action.

Eleven undescribed isoquinoline analogues, namely edulisines A-K, along with sixteen known alkaloids, were isolated from the whole plants of Corydalis edulis. The structures of the isolated alkaloids were established on the basis of extensive spectroscopic data (1D and 2D NMR, UV, IR, and HRESIMS). Their absolute configurations were determined by single-crystal X-ray crystallographic analysis and ECD. Compounds (+)-1 and (-)-1 are a pair of undescribed isoquinoline alkaloids bearing a unique coupled pattern of coptisine and ferulic acid via Diels-Alder [4 + 2] cycloaddition, while compounds (+)-2 and (-)-2 feature benzo [1,2-d:3,4-d]bis [1,3]dioxole moiety. Compounds (+)-2, (-)-2, (-)-5, 10, 13, 15, 20, 22, and 23 significantly triggered the secretion of insulin in the HIT-T15 cells at a concentration of 40 µM.

SMC1A regulated by KIAA1429 in m6A-independent manner promotes EMT progress in breast cancer.

As a component of N6-methyladenosine (m6A) "writers," KIAA1429 was reported to promote breast cancer proliferation and growth in m6A-independent manners. However, the related mechanism of KIAA1429 in breast cancer metastasis has not been reported. In the present study, we found KIAA1429 could significantly promote the migration and invasion of breast cancer cells. Then we demonstrated that knockdown of KIAA1429 could impede breast cancer metastasis in nude mice in vivo. The level of SNAIL expression and epithelial-mesenchymal transition (EMT) progress was positively related with KIAA1429. Furthermore, we confirmed that the suppression of cell migration, invasion, and EMT progress by knockdown of KIAA1429 could be reversed by the upregulation of SNAIL. However, structural maintenance of chromosomes 1A (SMC1A), not KIAA1429, bound with the SNAIL promoter region directly and promoted the transcription of SNAIL. Then we confirmed that KIAA1429 could bind to the motif in the 3' UTR of SMC1A mRNA directly and enhance SMC1A mRNA stability. In conclusion, our study revealed a novel mechanism of the KIAA1429/SMC1A/SNAIL axis in the regulation of metastasis of breast cancer. Moreover, it first provided detailed investigation of how KIAA1429 regulated the targeted gene expression at posttranscriptional levels as an RNA binding protein unrelated to its m6A modification.

[A new nor-sesquiterpene glycoside from Corydalis edulis].

This study is to investigate the chemical constituents from the whole plant Corydalis edulis. The chemical constituents were separated and purified by macroporous resin D101, silica gel, Sephadex LH-20, ODS, and semi-preparative HPLC. Their structures were determined by physicochemical properties and spectroscopic data. Four compounds were isolated from the dichloromethane and water extracts of the whole plant C. edulis, and identified as 6'-ß-D-xylosylicariside B2(1),(3S,5R,6S,7E)-5,6-epoxy-3-hydroxy-7-megastigmen-9-one(2), loliolide(3), and 5,5'-dimethoxybiphenyl-2,2'-diol(4), respectively. Compound 1 is a new compound, of which the absolute configuration was established by electronic circular dichroism(ECD) calculations. Compound 4 is obtained from the plants of Papaveraceae family for the first time. Compounds 2 and 3 are firstly isolated from the Corydalis genus.

[Open-top Chamber for in situ Research on Response of Mercury Enrichment in Rice to the Rising Gaseous Elemental Mercury in the Atmosphere].

In situ research was conducted on the response of mercury enrichment in rice organs to elevated gaseous elemental mercury (GEM) with open-top chambers (OTCs) fumigation experiment and soil Hg enriched experiment. The results showed that Hg concentrations in roots were generally correlated with soil Hg concentrations (R = 0.9988, P < 0.05) but insignificantly correlated with air Hg concentrations (P > 0.05), indicating that Hg in rice roots was mainly from soil. Hg concentrations in stems increased linearly (R(B) = 0.9646, R(U) = 0.9831, P < 0.05) with elevated GEM, and Hg concentrations in upper stems were usually higher than those in bottom stems in OTCs experiment. Hg concentrations in bottom stems were generally correlated with soil Hg concentrations (R = 0.9901, P < 0.05) and second-order polynomial (R = 0.9989, P < 0.05) was fitted for Hg concentrations in upper stems to soil Hg concentrations, and Hg concentrations in bottom stems were usually higher than those in upper stems in soil Hg enriched experiment, indicating the combining impact of Hg from air and soil on the accumulation of mercury in stems. Hg concentrations in foliage were significantly correlated (P < 0.05) with air Hg and linearly correlated with soil Hg (R = 0.9983, P = 0.0585), implying that mercury in foliage was mainly from air and some of Hg in root from soil was transferred to foliage through stem. Based on the function in these filed experiments, it was estimated that at least 60%-94% and 56%-77% of mercury in foliage and upper-stem of rice was from the atmosphere respectively, and yet only 8%-56% of mercury in bottom-stem was attributed to air. Therefore, mercury in rice aboveground biomass was mainly from the atmosphere, and these results will provide theoretical basis for the regional atmospheric mercury budgets and the model of mercury cycling.

Antiparasitic efficacy of Gracillin and Zingibernsis newsaponin from Costus speciosus (Koen ex. Retz) Sm. against Ichthyophthirius multifiliis.

The present study aims to evaluate the antiparasitic activity of active components from Costus speciosus against Ichthyophthirius multifiliis. Bioassay-guided fractionation was employed to identify active compounds from C. speciosus yielding 2 bioactive compounds: Gracillin and Zingibernsis newsaponin. In-vitro assays revealed that Gracillin and Zingibernsis newsaponin could be 100% effective against I. multifiliis at concentrations of 0.8 and 4.5 mg L(-1), with median effective concentration (EC50) values of 0.53 and 3.2 mg L(-1), respectively. All protomonts and encysted tomonts were killed when the concentrations of Gracillin and Zingibernsis newsaponin were 1.0 and 5.0 mg L(-1). In-vivo experiments demonstrated that fish treated with Gracillin and Zingibernsis newsaponin at concentrations of 1.0 and 5.0 mg L(-1) carried significantly fewer parasites than the control (P<0.05). Mortality of fish did not occur in the treatment group (Zingibernsis newsaponin at 5.0 mg L(-1)) during the trial, although 100% of untreated fish died. Acute toxicities (LD50) of Gracillin and Zingibernsis newsaponin for grass carp were 1.64 and 20.7 mg L(-1), respectively. These results provided evidence that the 2 compounds can be selected as lead compounds for the development of new drugs against I. multifiliis.

[Chemical constituents from the aerial part of Sibiraea angustata].

OBJECTIVE: To study the chemical constituents from the aerial part of Sibiraea angustata. METHODS: The constituents were isolated by various chromatographic techniques (HP-20 macroporous absorption resin, Sephadex LH-20 gel, RP-MPLC and PHPLC)and their structures were determined on the basis of physicochemical properties and their spectroscopic data,as well as literatures. RESULTS: Eleven compounds were separated and identified as p-methoxycinnamic acid(I), protocatechuic aldehyde(II), quercetin(III), isorhamnetin(IV), quercetin 3-O-beta-D-galactopyranoside (V),9-0-[beta-D-glucopyranoside]-3,4,5-trimethoxy cinnamyl alcohol(VI), syringaresinol-4'-O-beta-D-monoglucoside(VII), ntin(VIII), sibiraic acid(IX), sibiscolacton(X), methyl ferulic acid(XI). CONCLUSION: Compounds I-XIII are isolated from the genus of Sibiraea for the first time.

Eight New Monoterpene Acylglucosides from Sibiraea angustata.

Chemical investigation of the aqueous extract from the aerial part of Sibiraea angustata, has led to the isolation of eight new monoterpene acylglucosides named sibiraglycoside A-H(1-8), together with two known monoterpenes, Sibiraic acid (9) and Sibiskoside (10). Their structures were elucidated by means of extensive spectroscopic data analysis (including 1D and 2D NMR, HRESIMS experiments), as well as compared with the literature data. The relative configuration was established by NOE studies. In the in vitro bioassay, all the compounds showed moderate hypolipemic effects, among them, compounds 7 and 9, showed distinctive activity on lowering lipid.

Gaseous elemental mercury concentration in atmosphere at urban and remote sites in China.

An investigation of gaseous elemental mercury concentration in atmosphere was conducted at Beijing and Guangzhou urban, Yangtze Delta regional sites and China Global Atmosphere Watch Baseline Observatory (CGAWBO) in Mt. Waliguan of remote continental area of China. High temporal resolved data were obtained using automated mercury analyzer RA-915+. Results showed that the overall hourly mean Hg0 concentrations in Mt. Waliguan were 1.7+/-1.1 ng/m3 in summer and 0.6+/-0.08 ng/m3 in winter. The concentration in Yangtze Delta regional site was 5.4+/-4.1 ng/m3, which was much higher than those in Waliguan continental background area and also higher than that found in North America and Europe rural areas. In Beijing urban area the overall hourly mean Hg0 concentrations were 8.3+/-3.6 ng/m3 in winter, 6.5+/-5.2 ng/m3 in spring, 4.9+/-3.3 ng/m3 in summer, and 6.7+/-3.5 ng/m3 in autumn, respectively, and the concentration was 13.5+/-7.1 ng/m3 in Guangzhou site. The mean concentration reached the lowest value at 14:00 and the highest at 02:00 or 20:00 in all monitoring campaigns in Beijing and Guangzhou urban areas, which contrasted with the results measured in Yangtze Delta regional site and Mt. Waliguan. The features of concentration and diurnal variation of Hg0 in Beijing and Guangzhou implied the importance of local anthropogenic sources in contributing to the high Hg0 concentration in urban areas of China. Contrary seasonal variation patterns of Hg0 concentration were found between urban and remote sites. In Beijing the highest Hg0 concentration was in winter and the lowest in summer, while in Mt. Waliguan the Hg0 concentration in summer was higher than that in winter. These indicated that different processes and factors controlled Hg0 concentration in urban, regional and remote areas.