[Intestinal toxicity of n-BuOH fraction from Phytolacca Radix before and after being processed with vinegar].

To research the intestinal toxicity of n-BuOH fraction in Phytolacca Radix before and after being processed with vinegar. Toxic n-BuOH fractions were separated from Phytolacca Radix. In the animal model, the level of intestinal edema, water content of intestine and stool, IC50 values of HT-29 and IEC-6 were detected with MTT method to compare the changes in toxicity of n-BuOH fractions from Phytolacca Radix before and after being processed with vinegar. n-BuOH fractions of Phytolacca Radix could cause intestinal edema in mice, increase the edema of duodenum, jejunum and the water content in stool, inhibit the proliferation of HT-29 cells and IEC-6 cells, indicating its intestinal toxicity, with HT-29 IC50 at 14.59 mg•L⁻¹ and IEC-6 IC50 at 43.77 mg•L⁻¹. After being processed with vinegar, the level of intestinal edema, edema of duodenum and jejunum and the water content in stool and inhibition ratio of cells line were reduced, with HT-29 IC50 at 58.51 mg•L⁻¹ and IEC-6 IC50 at 84.37 mg•L⁻¹. After being processed with vinegar, the toxicity of n-BuOH fractions from Phytolacca Radix decreased obviously.

[Antagonism mechanism of gingerols against inflammatory effect of toxic raphides from Pinella pedatisecta].

This study was to investigate the mechanism of gingerols antagonizing the inflammatory effect of toxic raphides from Pinella pedatisecta. Mice peritonitis models induced by toxic raphides from P. pedatisecta were applied to observe the effect of gingerols on inflammatory mediators PGE2 in the exudates of abdominal inflammation in mice; rats peritoneal macrophage in vitro culture models were adopted to study the anti-inflammatory effects of gingerol against toxic raphides, with TNF-α and IL-1ß in supernatant as indexes. Scanning electron microscopy was used to observe the changes in surface morphology of macrophages treated by raphides and gingerols. Macrophages-neutrophils co-cultured models were used to study the antagonism of gingerols against the effect of toxic raphides' stimulation on neutrophils migration. Results showed that gingerols could significantly inhibit the production of PGE2 in the exudates of abdominal inflammation induced by toxic raphides from P. pedatisecta in mice. Gingerols could significantly inhibit the toxic raphides from P. pedatisecta to induce the release of inflammatory factors, with certain dose dependence. Scanning electron microscopy showed that gingerols could significantly inhibit phagocytosis of macrophages, cytomembrane injury, and neutrophils migration induced by toxic raphides from P. pedatisecta. The results showed that the antagonism mechanism of gingerols against the toxic raphides from P. pedatisecta may be associated with inhibiting the pro-inflammatory toxicity including macrophage activation, inflammatory factors release, and neutrophils migration.

Tracing novel hemostatic compounds from heating products of total flavonoids in Flos Sophorae by spectrum-effect relationships and column chromatography.

Flos Sophorae and its processed product have been clinically used to treat hemorrhage. In this study, the total ion chromatographic fingerprints of the heating products of total flavonoids in Flos Sophorae were established by high-performance liquid chromatography with tandem mass spectrometry and the hemostatic activities were studied by hemostatic screening tests in vivo. The spectrum-effect relationships between fingerprints and hemostatic activities were investigated using canonical correlation analysis to trace the peaks responsible for the hemostatic effects. The predicted active peaks in fingerprints were isolated by column chromatography and their structures were identified by NMR spectroscopy and mass spectrometry. The hemostatic activities of them were verified by platelet aggregation and procoagulation assays in vitro. Canonical correlation analysis results showed that peak 8 and peak 11 were correlated most closely, thus probably being the main hemostatic compounds. Through column chromatography separation, peak 8 (compound I) and peak 11 (compound II) were obtained with purities of 95.61 and 93.38%, respectively, and were discovered new hemostatic compounds named as huaicarbon A (I) and huaicarbon B (II), respectively. This study provides a universal model to trace the active compounds of other herbs which have bioactivity enhancement after processing by spectrum-effect relationships and column chromatography.

A Novel Reduplicate Strategy for Tracing Hemostatic Compounds from Heating Products of the Flavonoid Extract in Platycladi cacumen by Spectrum-Effect Relationships and Column Chromatography.

Platycladi cacumen and its processed product have been utilized as a Chinese medicine to treat hemorrhages. In this study, the base peak chromatogram fingerprints of heating products of total flavonoids in Platycladi cacumen were established by high performance liquid chromatography coupled with mass spectroscopy/mass spectroscopy (HPLC-MS/MS), and the hemostatic activities were studied by hemostatic screening tests in vivo. The spectrum-effect relationships between fingerprints and hemostatic activities were analyzed by using canonical correlation analysis to trace the peaks responsible for the significant hemostatic effects. Peak 10 and peak 12 were correlated most closely, thus probably being the main hemostatic compounds. To confirm the reliability of this strategy, the targeted unknown peak was obtained by bioactivity-guided isolation, characterized by MS, ¹H-NMR, (13)C-NMR, and 2D-NMR spectroscopies, and referred to as cecarbon as a new compound. In addition, the isolated compound exhibited hemostatic effect in a dose-dependent manner with different potencies in vitro and existed in Platycladi cacumen Carbonisatus. A novel dereplication strategy was employed to trace and identify the active compounds of other herbs that have bioactivity enhancement after processing using spectrum-effect relationships and column chromatography.

Characterization and Quantification by LC-MS/MS of the Chemical Components of the Heating Products of the Flavonoids Extract in Pollen Typhae for Transformation Rule Exploration.

The Traditional Chinese Medicine herbs Pollen Typhae and Pollen Typhae Carbonisatus have been used as a hemostatic medicine promoting blood clotting for thousands of years. In this study, a reliable, highly sensitive method based on LC-MS/MS has been developed for differentiation of the heating products of total flavonoids in Pollen Typhae (FPT-N). Twenty three peaks were detected and 18 peaks have been structurally identified by comparing retention times, high resolution mass spectrometry data, and fragment ions with those of the reference substances and/or literature data. Additionally, 15 compounds have been quantified by multiple reaction monitoring in the negative ionization mode. It was found that the contents of the characterized compounds differed greatly from each other in FPT-N samples. Among them, the content of huaicarbon B significantly increased at first, while it decreased after heating for 25 min, which could be considered as the characteristic component for distinguishing FPT-N. The present study provided an approach to rapidly distinguish the differences of FPT-N samples. In addition, the actively summarized characteristic fragmentation might help deducing the structure of unknown flavonols compounds. Furthermore, transformation rules of flavonoids during the heating process in carbonisatus development could contribute to hemostatic therapeutic component exploration.

[Antagonistic effect of gingerols against TNF-α release, ROS overproduction and RIP3 expression increase induced by lectin from Pinellia ternata].

To explore the antagonistic effect of gingerols against the inflammation induced by lectin from Pinellia ternata. In this study, ELISA method was used to determine the effect of different extracts from gingerols on the release of inflammatory factor TNF-α from macrophages induced by lectin from P. ternata. The fluorescence probe was used to determine the effect of gingerols on the changes in ROS of macrophages induced by lectin from P. ternata. The western-blot method was applied to study the effect of gingerols on the increase in expression of cell receptor interacting protein RIP3 in macrophages induced by lectin from P. ternata. The scanning electron microscope (SEM) was used to study the effect of gingerols on morphological changes in macrophages induced by lectin from P. ternata. According to the results, gingerols can significantly inhibit the release of inflammatory factor from macrophages induced by lectin from P. ternata, ROS overproduction and increase in RIP3 expression. SEM results showed that gingerols can inhibit the cytomorphosis and necrocytosis induced by lectin from P. ternata. Fresh ginger's detoxication may be related to gingerols' effects in inhibiing release of inflammatory factor, ROS overproduction and increase in RIP3 expression caused by macrophages induced by lectin from P. ternata, which are mainly inflammatory development.

[Totoxicity fraction from Euphorbia pekinensis and composition change after vinegar processing].

To look for the toxicity fraction of Euphorbia pekinensis and discuss the vinegar processing mechanism. The level of intestinal edema, water content of intestine and stool, IC50 values of IEC-6 were applied to evaluate the toxicity of different fractions. RT-PCR was employed for detecting AQP1, AQP3 mRNA expression. The petroleum ether (PE) fraction and ethyl acetate (EtOAc) fraction could significant cause intestinal edema in mice, increase the water content of duodenum, colon and stool, inhibited the mRNA expression of AQP1 and increased the mRNA level of AQP3 in colon, and the petroleum ether (PE) fraction was more poisonous. After the petroleum ether (PE) fraction was processed with vinegar, the level of intestinal edema, water content of duodenum, colon, stool and inhibition ratio of cells line were reduced. And we compared the composition change after vinegar processing, finding that the conpekinensis.

Lotus Metasurface for Wide-Angle Intermediate-Frequency Water-Air Acoustic Transmission.

Only 0.1% of the acoustic energy can transmit across the water-air interface because of the huge acoustic impedance mismatch. Enhancing acoustic transmission across the water-air interface is of great significance for sonar communications and sensing. However, due to the interface instability and subwavelength characteristics of acoustic metamaterials, wide-angle intermediate-frequency (10 kHz-100 kHz) water-air acoustic transmission remains a great challenge. Here, we demonstrate that the lotus leaf is a natural low-cost acoustic transmission metasurface, namely, the lotus acoustic metasurface (LAM). Experiments demonstrate the LAM can enhance the acoustic transmission across the water-air interface, with an energy transmission coefficient of about 40% at 28 kHz. Furthermore, by fabricating artificial LAMs, the operating frequencies can be flexibly adjusted. Also, the LAM allows a wide-angle water-to-air acoustic transmission. It will enable various promising applications, such as detecting and imaging underwater objects from the air, communicating between ocean and atmosphere, reducing ocean noises, etc.

Tunable Fluid-Type Metasurface for Wide-Angle and Multifrequency Water-Air Acoustic Transmission.

Efficient acoustic communication across the water-air interface remains a great challenge owing to the extreme acoustic impedance mismatch. Few present acoustic metamaterials can be constructed on the free air-water interface for enhancing the acoustic transmission because of the interface instability. Previous strategies overcoming this difficulty were limited in practical usage, as well as the wide-angle and multifrequency acoustic transmission. Here, we report a simple and practical way to obtain the wide-angle and multifrequency water-air acoustic transmission with a tunable fluid-type acoustic metasurface (FAM). The FAM has a transmission enhancement of acoustic energy over 200 times, with a thickness less than the wavelength in water by three orders of magnitude. The FAM can work at an almost arbitrary water-to-air incident angle, and the operating frequencies can be flexibly adjusted. Multifrequency transmissions can be obtained with multilayer FAMs. In experiments, the FAM is demonstrated to be stable enough for practical applications and has the transmission enhancement of over 20 dB for wide frequencies. The transmission enhancement of music signal across the water-air interface was performed to demonstrate the applications in acoustic communications. The FAM will benefit various applications in hydroacoustics and oceanography.

Typhonium giganteum Lectin Exerts A Pro-Inflammatory Effect on RAW 264.7 via ROS and The NF-κB Signaling Pathway.

Typhonii rhizoma, a widely used herb in traditional Chinese medicine, has acute irritating toxicity related to Typhonium giganteum lectin (TGL). TGL exhibits acute inflammatory effects, but the underlying molecular mechanisms are largely unknown. This paper is designed to assess the pro-inflammatory response of TGL on RAW 264.7 cells. RAW 264.7 treated with 6.25, 12.5, 25, and 50 µg/mL TGL showed elevated levels of inflammatory factors (TNF-α, IL-1ß) and of p-IκB and p-p65, all dose-dependent, indicating that TGL had a substantial inflammatory effect and mobilized the nuclear factor-κB (NF-κB) pathway. All four TGL treatments also induced the up-regulation of reactive oxygen species (ROS) and cytosolic free Ca2+ and down-regulation of mitochondrial membrane potential (MMP). The production of cytokines and p-IκB, p-p65 were reduced by N-acetylcysteine (NAC), an ROS scavenger, which somewhat abrogated ROS production. The results showed the TGL-activated inflammatory signaling pathway NF-κB to be associated with the overproduction of ROS. Moreover, 50 µg/mL treatment with TGL led to cell apoptosis after 1 h and increased necrosis over time. These results provided potential molecular mechanisms for the observed inflammatory response to TGL including up-regulation of ROS and cytosolic free Ca2+, down-regulation of MMP, the mobilization of the NF-κB pathway, and the subsequent overproduction of pro-inflammatory factors resulting in apoptosis. Long-term stimulation with TGL resulted in strong toxic effects related to inflammation that induced necrosis in macrophages.

Toxic proteins from Croton tiglium L. exert a proinflammatory effect by inducing release of proinflammatory cytokines and activating the p38-MAPK signaling pathway.

The aim of the present study was to determine the toxic targets of proteins from Croton tiglium L. and to investigate the potential mechanism of their toxicity. The toxic targets were determined by oral medication and intraperitoneal injection. The median lethal dose of oral medication in mice was calculated using Bliss software (2,752.8-3,407.5 mg/kg), and that of intraperitoneal injection was 195.8­272.69 mg/kg. The results of histopathological examination demonstrated that the kidney was primarily impaired by intraperitoneal injection, with slight degeneration of renal tubular epithelial cells. As to oral medication, the digestive tract was primarily injured, which manifested as congestion, bleeding, serious edema and other symptoms. Oral administration of the proteins caused gastrointestinal edema by increasing the intestinal permeability. Severe edema was associated with the inflammatory response, therefore the association between the toxicity of the proteins and inflammation was investigated. The proinflammatory effects of the crude proteins on the release of inflammatory mediator prostaglandin E2 (PGE2) were evaluated through intraperitoneal injection and the production of proinflammatory cytokines in RAW264.7 macrophages. Maximum PGE2 was released in the mice in vivo following intraperitoneal injection with 400 mg crude protein/kg body weight. Proinflammatory cytokines in macrophages, including tumor necrosis factor­α and interleukin­1ß, were produced in dose­ and time­dependent manners in vitro. furthermore, the expressions of cell signaling molecules were detected by western blotting. The inflammatory response induced by crude protein in macrophages was associated with the mitogen­activated protein kinase (MAPK) signaling pathway mainly including p38­MAPK, extracellular signal­regulated kinase 1/2 and c­Jun N­terminal kinase 1/2/3 and the activated p38­MAPK signaling pathway. However, extracellular signal­regulated kinase 1/2 and c­Jun N­terminal kinases 1­3 exhibited no significant response.

Correlation between proinflammatory role of a lectin from Typhonium giganteum Engl. and macrophage.

PURPOSE: To analyze the correlation between proinflammatory effects of a lectin from Typhonium giganteum Engl. and macrophage. METHODS: T. giganteum lectin (TGL) was extracted from the tuber of T. giganteum and purified, and was then identified by using SDS-PAGE gel electrophoresis in combination with mass spectrometry. The morphologic changes of macrophage after being stimulated by TGL were observed with scanning electron microscopy. The influences of such stimulation on neutrophil migration were evaluated by establishing an in vitro macrophage-neutrophil co-culture migration model. By establishing a rat peritoneal macrophage in vitro cultured model, the effects of TGL stimulation on inflammatory factors TNF-α and IL-1ß released by macrophage were analyzed. With p65 as the index, the expressions of the NF-κB signaling pathway in the cytoplasm and nucleus were detected before and after TGL stimulation respectively. Furthermore, we also investigated whether the inhibitor for NF-κB signaling pathway BAY11-7082 can block p65 nuclear translocation. RESULTS: After being stimulated by TGL, macrophage had increased volume, number of pseudopodia and gradually cracked cell membrane, accompanied by evidently induced migration of neutrophils due to released inflammatory factors. As the concentration of TGL varied, NF-κB's monomer p65 had different expression levels in the cytoplasm and nucleus, while BAY11-7082 can indeed block the nuclear translocation of p65. CONCLUSIONS: TGL-induced inflammation was closely related to macrophage mediation.

The alum-processing mechanism attenuating toxicity of Araceae Pinellia ternata and Pinellia pedatisecta.

UNLABELLED: The present study aimed at investigating the alum-processing mechanism attenuating toxicity of Araceae Pinellia ternata and Pinellia pedatisecta. Animal retroperitoneal inflammatory model in vivo and macrophagocyte release inflammatory factor model in vitro were used to detect the effect of alum processing on raphides and lectin. Scanning electron microscopy was used to observe the change in raphides during processing; HPLC method was used to determine the correlation between the dissolution and corrosion of raphides and ion in the alum solution; (27)Al-NMR technology was used to detect the relationship between aluminum oxalate complex formation and the dissolved and corrosion of raphides. The change in protein peptide sequence of lectin during the processing of alum solution was determined by Shotgun LC-MS assay. Raphides induced severe rabbit conjunctival edema and an intraperitoneal injection of lectin increased PGE2 and protein in mice peritoneal exudate, while decreased after treatment with alum solution processing. During the processing raphides was dissolved and corroded, then its structure was damaged. Raphides was soaked in the alum solution and significantly decreased the oxalate content, and the effect was related with Al(3+) in the alum. Al(3+) in the alum combined with C2O4(2-) of raphides into a stable complex compound promoted the dissolution of calcium oxalate. Raphides soaked in the alum made lectin proteins dissolve, whereas protein peptide sequence of lectin was changed and the protein structure was damaged. CONCLUSION: Alum solution could decrease the toxicity of P. ternata (Thunb.) Breit. and P. pedatisecta Schott. Since it made a special crystal structure of raphides damage and the protein of lectin dissolve. The structure of toxic substances significantly changed, which decreased the inflammatory effect.

Pinellia ternata lectin exerts a pro-inflammatory effect on macrophages by inducing the release of pro-inflammatory cytokines, the activation of the nuclear factor-κB signaling pathway and the overproduction of reactive oxygen species.

Pinellia ternata (PT) is a widely used traditional Chinese medicine. The raw material has a throat-irritating toxicity that is associated with the PT lectin (PTL). PTL is a monocot lectin isolated from the tubers of PT, which exhibits mouse peritoneal acute inflammatory effects in vivo. The present study aimed to investigate the pro-inflammatory effect of PTL on macrophages. PTL (50 µg/ml)­stimulated macrophages enhanced the chemotactic activity of neutrophils. PTL (50, 100, 200 and 400 µg/ml) significantly elevated the production of cytokines [tumor necrosis factor­α (TNF-α) , interleukin (IL)­1ß and IL­6]. PTL (25, 50 and 100 µg/ml) induced intracellular reactive oxygen species (ROS) overproduction. PTL also caused transfer of p65 from the macrophage cytoplasm to the nucleus and activated the nuclear factor­κB (NF­κB) signaling pathway. Scanning electron microscope images revealed severe cell swelling and membrane integrity defection of macrophages following PTL (100 µg/ml) stimulation, which was also associated with inflammation. PTL had pro­inflammatory activity, involving induced neutrophil migration, cytokine release, ROS overproduction and the activation of the NF-κB signaling pathway, which was associated with the activation of macrophages.

A new casbane diterpene from Euphorbia pekinensis.

A new casbane diterpenoid, referred to as pekinenin G, together with one cembrane diterpene and four known casbane diterpenoids were isolated from the roots of Euphorbia pekinensis. Their structures were elucidated on the basis of spectroscopic studies and comparison with related known compounds. The six compounds showed different cytotoxic activities against four human cancer cell lines.