Chemicolome and Metabolome Profiling of Xieriga-4 Decoction, A Traditional Mongolian Medicine, Using the UPLC-QTOF/MS Approach.

Background: Xieriga-4 decoction (XRG-4) is a classic prescription Mongolian medicine that has potent diuretic and anti-inflammatory activities. However, its functional components remain unknown. Purpose: This study aimed to identify the chemical components in XRG-4 and its metabolome in vivo. Methods: An ultra-performance liquid chromatography coupled with a quadrupole time-of-flight tandem mass spectrometry based approach was proposed to systematically profile the chemicolome and metabolome of XRG-4. Result: A total of 106 constituents were identified in XRG-4. Eighty-nine components were identified in biological samples, including 78 in urine (24 prototypes and 54 metabolites), 26 in feces (19 prototypes and 7 metabolites), and 9 in plasma (5 prototypes and 4 metabolites). In other tissues, only a few compounds, including alkaloids and iridoids, were detected. Conclusion: This comprehensive investigation of the chemical and metabolic profiles of XRG-4 provides a scientific foundation for its quality control and administration of clinically-safe medication.

Genipin improves lipid metabolism and sperm parametersin obese mice via regulation of miR-132 expression.

Obesity has now surpassed malnutrition and infectious diseases as the most significant contributor to health problems worldwide. In particular, obesity is associated with several metabolic disorders, including hyperlipidemia, hepatic steatosis, and subfertility. Genipin (GNP), the aglycone of geniposide, is isolated from the extract of the traditional Chinese medicine Gardenia jasminoides Ellis and has been used in traditional oriental medicine against several inflammation-driven diseases. However, the effect and molecular mechanism of GNP on obesity-associated dyslipidemia and sperm dysfunction still need to be explored. In this study, we detect the effects of GNP on hyperlipidemia, hepatic lipid accumulation and sperm function using a high-fat diet (HFD)-induced obese mouse model. We find that obese mice treated with GNP show an improvement in body weight, serum triglyceride levels, serum hormone levels, serum inflammatory cytokines, hepatic steatosis and sperm function. At the molecular level, HFD/GNP diversely regulates the expression of miR-132 in a tissue-specific manner. miR-132 further targets and regulates the expression of SREBP-1c in liver cells, as well as the expressions of SREBP-1c and StAR in Leydig cells in the testis, thus modifying lipogenesis and steroidogenesis, respectively. Collectively, our data demonstrate that GNP shows a broad effect on the improvement of HFD-induced metabolic disorder and sperm dysfunction in male mice by tissue-specific regulation of miR-132. Our findings reveal the function GNP in ameliorating hepatic lipid metabolism and sperm function and suggest that this compound is a versatile drug to treat metabolic disorders.

METTL3 antagonizes 5­FU chemotherapy and confers drug resistance in colorectal carcinoma.

Colorectal cancer (CRC) is one of top five leading causes of cancer­associated mortalities worldwide. 5­Fluorouracil (5­FU) is the first­line chemotherapeutic drug in the treatment of CRC; however, its antineoplastic efficiency is limited due to acquired drug resistance. The regulatory mechanism underlying 5­FU chemotherapeutic response and drug resistance in CRC remains largely unknown. The present study identified that silencing of methyltransferase­like 3 (METTL3) suppressed the proliferation and migration of CRC HCT­8 cells. Using cell survival assays, flow cytometric and colony formation analyses, it was revealed that inhibition of METTL3 sensitized HCT­8 cells to 5­FU by enhancing DNA damage and inducing apoptosis in HCT­8 cells under 5­FU treatment. Furthermore, the expression of METTL3 was upregulated in 5­FU­resistant CRC cells (HCT­8R), which contributed to drug resistance through regulation of RAD51 associated Protein 1 (RAD51AP1) expression. Western blotting, immunofluorescence staining and drug sensitivity assays demonstrated that knockdown of METTL3 augmented 5­FU­induced DNA damage and overcame 5­FU­resistance in HCT­8R cells, which could be mimicked by inhibition of RAD51AP1. The present study revealed that the METTL3/RAD51AP1 axis plays an important role in the acquisition of 5­FU resistance in CRC, and targeting METTL3/RAD51AP1 may be a promising adjuvant therapeutic strategy for patients with CRC, particularly for those with 5­FU­resistant CRC.

Deficiency of proline/serine-rich coiled-coil protein 1 (PSRC1) accelerates trimethylamine N-oxide-induced atherosclerosis in ApoE-/- mice.

AIMS: The main therapeutic strategies for coronary artery disease (CAD) are mainly based on the correction of abnormal cholesterol levels; however, residual risks remain. The newly proven gut microbial metabolite trimethylamine N-oxide (TMAO) linked with CAD has broadened our horizons. In this study, we determined the role of proline/serine-rich coiled-coil protein 1 (PSRC1) in TMAO-driven atherosclerosis. METHODS AND RESULTS: We first analyzed the levels of TMAO and PSRC1 in patients with or without atherosclerosis with a target LDL-C < 1.8 mmol/L. Plasma TMAO levels were increased and negatively associated with decreased PSRC1 in peripheral blood mononuclear cells. Animals and in vitro studies showed that TMAO inhibited macrophage PSRC1 expression due to DNA hypermethylation of CpG islands. ApoE-/- mice fed a choline-supplemented diet exhibited reduced PSRC1 expression accompanied by increased atherosclerotic lesions and plasma TMAO levels. We further deleted PSRC1 in apoE-/- mice and PSRC1 deficiency significantly accelerated choline-induced atherogenesis, characterized by increased macrophage infiltration, foam cell formation and M1 macrophage polarization. Mechanistically, we overexpressed and knocked out PSRC1 in cultured macrophages to explore the mechanisms underlying TMAO-induced cholesterol accumulation and inflammation. PSRC1 deletion impaired reverse cholesterol transport and enhanced cholesterol uptake and inflammation, while PSRC1 overexpression rescued the proatherogenic phenotype observed in TMAO-stimulated macrophages, which was partially attributed to sulfotransferase 2B1b (SULT2B1b) inhibition. CONCLUSIONS: Herein, clinical data provide evidence that TMAO may participate in the development of CAD beyond well-controlled LDL-C levels. Our work also suggests that PSRC1 is a negative regulator mediating the unfavorable effects of TMAO-containing diets. Therefore, PSRC1 overexpression and reduced choline consumption may further alleviate atherosclerosis.

Deficiency of PSRC1 accelerates atherosclerosis by increasing TMAO production via manipulating gut microbiota and flavin monooxygenase 3.

Maladaptive inflammatory and immune responses are responsible for intestinal barrier integrity and function dysregulation. Proline/serine-rich coiled-coil protein 1 (PSRC1) critically contributes to the immune system, but direct data on the gut microbiota and the microbial metabolite trimethylamine N-oxide (TMAO) are lacking. Here, we investigated the impact of PSRC1 deletion on TMAO generation and atherosclerosis. We first found that PSRC1 deletion in apoE-/- mice accelerated atherosclerotic plaque formation, and then the gut microbiota and metabolites were detected using metagenomics and untargeted metabolomics. Our results showed that PSRC1 deficiency enriched trimethylamine (TMA)-producing bacteria and functional potential for TMA synthesis and accordingly enhanced plasma betaine and TMAO production. Furthermore, PSRC1 deficiency resulted in a proinflammatory colonic phenotype that was significantly associated with the dysregulated bacteria. Unexpectedly, hepatic RNA-seq indicated upregulated flavin monooxygenase 3 (FMO3) expression following PSRC1 knockout. Mechanistically, PSRC1 overexpression inhibited FMO3 expression in vitro, while an ERα inhibitor rescued the downregulation. Consistently, PSRC1-knockout mice exhibited higher plasma TMAO levels with a choline-supplemented diet, which was gut microbiota dependent, as evidenced by antibiotic treatment. To investigate the role of dysbiosis induced by PSRC1 deletion in atherogenesis, apoE-/- mice were transplanted with the fecal microbiota from either apoE-/- or PSRC1-/-apoE-/- donor mice. Mice that received PSRC1-knockout mouse feces showed an elevation in TMAO levels, as well as plaque lipid deposition and macrophage accumulation, which were accompanied by increased plasma lipid levels and impaired hepatic cholesterol transport. Overall, we identified PSRC1 as an atherosclerosis-protective factor, at least in part, attributable to its regulation of TMAO generation via a multistep pathway. Thus, PSRC1 holds great potential for manipulating the gut microbiome and alleviating atherosclerosis.

The protective role and mechanism of liriodendrin in rats with myocardial infarction.

BACKGROUND: Liriodendrin is a therapeutic constituent of sargentgloryvine stem which is a famous Chinese traditional medicine. Previous studies have suggested liriodendrin could inhibit different pathways to treat inflammation in lung and intestinal tract. But whether it can treat myocardial infarction (MI) is unknown. We investigated the protective effect of liriodendrin on acute MI in rats and explored the specific mechanisms to expand the use of this traditional Chinese medicine. METHODS: The rats were randomized into the sham group (sham operation), control group (ligation of the left anterior descending artery), and liriodendrin group. The liriodendrin group was intragastrically administered with a liriodendrin solution (100 mg/kg). The control group and the sham group were intragastrically administered with normal saline. Before all rats were euthanized, echocardiography was used to detect their cardiac function. Hematoxylin and eosin (HE) staining and the terminal deoxynucleotidyl transferase-mediated nick end labeling (TUNEL) method were performed. Further quantitative detection of interleukin-1ß (IL-1ß) and tumor necrosis factor-α (TNF-α) levels in tissues were also detected. Western Blot and real-time polymerase chain reaction (RT-PCR) were used to detect the apoptosis and nuclear factor kappa-B (NF-κB) pathway in tissues. H9C2 cells were used to detect the related mechanisms in vitro. RESULTS: Echocardiography showed that, compared to control group, the cardiac function of the liriodendrin group was significantly improved. histopathological staining of the control group showed that the myocardial tissue was severely damaged, and inflammatory cells were infiltrated. Compared to the control group, the apoptosis index of the liriodendrin group was significantly lower (P<0.05). Enzyme-linked immunosorbent assay (ELISA) results showed that the levels of IL-1ß and TNF-α in the control group were higher than those in the liriodendrin group (P<0.05). Meanwhile, apoptosis and the NF-κB pathway were inhibited after liriodendrin administration (P<0.05). Moreover, the mRNA transcriptional activity in the control group was also higher than that in the liriodendrin group (P<0.05). Because of the effect of liriodendrin, NF-κB pathway and apoptosis were downregulated in H9C2 cells which were exposed to ischemia-hypoxia. CONCLUSIONS: Liriodendrin may protect myocardial cells after myocardial infarction in rats by inhibiting the release of inflammatory factors, activation of the NF-κB pathway, and apoptosis.

Quercetin Relieves the Excised Great Saphenous Vein Oxidative Damage and Inflammatory Reaction.

OBJECTIVE: The patency and quality of transplanted great saphenous vein (GSV) can seriously influence the physical state and life quality of patients who accepted the coronary artery bypass grafting (CABG). Quercetin is known for antioxidant, antithrombotic, anti-inflammatory, and antitumor properties. In this study, we examined the protection of quercetin to the great saphenous vein from oxidative and inflammatory damage. METHODS: The GSVs were collected from 15 patients undergoing CABG and cultured. Treated the veins by H2O2 and detected the NO, SOD, and MDA content by the relevant kits to explore the quercetin protection against oxidative damage. Then, for another group of GSVs, sheared them and detected the inflammatory cytokines, such as IL-6, TNFα, CCL20, PCNA, and VEGF. Collect the veins for H&E staining and PCNA and VEGF immunofluorescent staining. RESULTS: Pretreatment by quercetin reduced the production of NO and MDA induced by H2O2, and increased SOD activity. Quercetin also supressed the mRNA expressions of IL-6, TNFα after mechanical damage and had no influence on CCL20 and VEGF. Consistent with the lower expression of PCNA treated by quercetin, the vein intima was thinner. CONCLUSION: These results demonstrated that quercetin protects GSVs by reducing the oxidative damage and inflammatory response and also suppresses the abnormal thickening of venous endothelium by inhibiting cell proliferation. It reminded that, to some extent, quercetin has the potential to release the great saphenous vein graft damage.

Inhibition of miR-1193 leads to synthetic lethality in glioblastoma multiforme cells deficient of DNA-PKcs.

Glioblastoma multiforme (GBM) is the most malignant primary brain tumor and has the highest mortality rate among cancers and high resistance to radiation and cytotoxic chemotherapy. Although some targeted therapies can partially inhibit oncogenic mutation-driven proliferation of GBM cells, therapies harnessing synthetic lethality are 'coincidental' treatments with high effectiveness in cancers with gene mutations, such as GBM, which frequently exhibits DNA-PKcs mutation. By implementing a highly efficient high-throughput screening (HTS) platform using an in-house-constructed genome-wide human microRNA inhibitor library, we demonstrated that miR-1193 inhibition sensitized GBM tumor cells with DNA-PKcs deficiency. Furthermore, we found that miR-1193 directly targets YY1AP1, leading to subsequent inhibition of FEN1, an important factor in DNA damage repair. Inhibition of miR-1193 resulted in accumulation of DNA double-strand breaks and thus increased genomic instability. RPA-coated ssDNA structures enhanced ATR checkpoint kinase activity, subsequently activating the CHK1/p53/apoptosis axis. These data provide a preclinical theory for the application of miR-1193 inhibition as a potential synthetic lethal approach targeting GBM cancer cells with DNA-PKcs deficiency.

Comparison of PM2.5 carbonaceous pollutants between an urban site in Shanghai and a background site in a coastal East China Sea island in summer: concentration, composition and sources.

Nine paired samples of atmospheric particulate matter with an aerodynamic diameter less than or equal to 2.5 µm (PM2.5) were collected concurrently from an urban site in Shanghai, China and a background site in Huaniao Island (HNI) in the coastal East China Sea (ECS) between July 21 and 29, 2011. The samples were analyzed for 16 United States Environmental Protection Agency (USEPA) priority polycyclic aromatic hydrocarbons (PAHs), n-alkanes (20 species, C14-C33), hopanes (10 species, C29-C32), and steranes (12 species, C27-C29). These two sites, approximately 66 km apart, are both on the pathway of land-based pollutants as they are transported to the ECS by seasonal winds. As expected, concentrations in Shanghai were higher (average: 8.4 and 67.8 ng m-3 for the 16 PAHs and n-alkanes, respectively) than those in HNI (average: 1.8 and 8.5 ng m-3, respectively). The dominant contributor to the 16 PAHs in Shanghai was 5-6-ring PAHs (60.0%), whereas 2-3-ring PAHs contributed the most (72.5%) in HNI. Plant waxes contributed 45.7% and 25.9% of the n-alkanes in Shanghai and HNI, respectively, implying a relatively greater contribution from petroleum residues to the n-alkanes in HNI. Principal component analysis (PCA) and the compositions of hopanes and steranes highlighted a prominent contribution from traffic emissions to carbonaceous PM2.5 aerosols. This study provides comprehensive details about the sources, formation, and transport of pollutants from eastern China to the coastal ECS.

Seasonal variation of carbonaceous pollutants in PM2.5 at an urban 'supersite' in Shanghai, China.

Multiple PM2.5 samples collected through different seasons from October 2011 to August 2012 at an urban site in Shanghai, China were analyzed for carbonaceous pollutants. Data from this site, a 'super' air quality monitoring station at Fudan University, has been used by researchers to investigate the formation mechanism of haze episodes. The characteristics and concentrations of organic carbon (OC), elemental carbon (EC), n-alkanes, as well as relative abundances of hopanes, in these samples were determined. The concentrations showed a pronounced annual cycle with higher values in cold seasons (spring and winter, mean: 8.6 µg/m(3), 3.3 µg/m(3) and 136.4 ng/m(3) for OC, EC and n-alkanes, respectively) and lower values in warm seasons (fall and summer, mean: 6.6 µg/m(3), 2.6 µg/m(3) and 73.8 ng/m(3) for OC, EC and n-alkanes, respectively). EC generally displayed a common source with that of OC in all seasons. Petroleum residue was the major source of n-alkanes, contributing 71.4% to the targeted C14-C33n-alkanes over four seasons. Principal components analysis and the composition of hopanes showed that emissions from vehicle exhaust contributed more carbonaceous aerosols than coal combustion. These data could provide important information for measures to reduce carbonaceous pollutant emissions and improve air quality in Shanghai, and other urban centers across China.

Protective effect of Xingnaojia formulation on rats with brain and liver damage caused by chronic alcoholism.

The aim of this study was to observe the effect of a formulation of traditional Chinese medicine extracts known as Xingnaojia (XNJ) on the liver function, learning ability and memory of rats with chronic alcoholism and to verify the mechanism by which it protects the brain and liver. A rat model of chronic alcoholism was used in the study. The spatial learning ability and memory of the rats were tested. The rats were then sacrificed and their brains and hepatic tissues were isolated. The activity of superoxide dismutase (SOD) and levels of glutamate (Glu), N-methyl D-aspartate receptor subtype 2B (NR2B), cyclin-dependent kinase 5 (CDK5) and cannabinoid receptor 1 (CB1) in the hippocampus were analyzed. The ultrastructure of the hepatic tissue was observed by electron microscopy. In addition, the activities of alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) in serum were tested and the levels of low-density lipoprotein (LDL), high-density lipoprotein (HDL), triglycerides (TG) and total cholesterol (TCHOL) were analyzed. XNJ enhanced the learning and memory of rats with chronic alcoholism. Treatment with XNJ increased the activity of SOD, and decreased the expression levels of NR2B mRNA and NR2B, CB1 and CDK5 proteins in the brain tissues compared with those in the model rats. It also increased the activity of ALDH in the serum and liver, decreased the serum levels of LDL, TG and TCHOL and increased the serum level of HDL. These results indicate that XNJ exhibited a protective effect against brain and liver damage in rats with chronic alcoholism.

Sources and preservation of organic matter in soils of the wetlands in the Liaohe (Liao River) Delta, North China.

Total organic carbon, total nitrogen, δ(13)Corg, δ(15)N, and aliphatic and polyaromatic hydrocarbons of fifty-five soil samples collected from the coastal wetlands of the Liaohe Delta were measured, in order to determine the sources and possible preservation of organic matter (OM). The δ(15)N and δ(13)Corg values in the samples ranged from 3.0‰ to 9.4‰ and from -30.4‰ to -20.3‰, respectively, implying that the OM in the soils is predominantly derived from C3 plant. The long-chain n-alkanes had a strong odd-over-even carbon number predominance, suggesting a significant contribution from waxes of higher plants. The ubiquitous presence of unresolved complex mixture, alkylated polycylic aromatic hydrocarbons and typical biomarkers of petroleum hydrocarbons (pristane, phytane, hopanes and steranes) indicates that there is a contribution of petroleum hydrocarbons to the organic carbon pool in the wetland soils. P. australis-vegetated wetlands have strong potentials for the preservation of organic carbon in the wetlands.

[Comparison on ingredients between cultured cell and cultivated Saffron pistils].

OBJECTIVE: To compare the differences of active ingredients between tissue cultured cells and cultivated saffron pistils. METHOD: The experiment was carried out by the Fourier transform infrared spectroscopy (FTIR) spectra and high performance liquid chromatography (HPLC). RESULT: The data indicated that the species and contents active ingredients in saffron pistils from different places were different. The species of active ingredients in tissue cultured cells are less than those in cultivated saffron pistils. However, the quantity of crocin A, which showed good anticancer effect, is 2-3 times more than that in cultivated saffron pistils. CONCLUSION: The active ingredients of the tissue cultured cells are similar to those of saffron pistils, but their contents are different. Therefore, the tissue cultured cells can only be the part-substitutes of cultivated saffron pistils.

[Analysis of contents of berberine in Coptis chinensis of Lichuan].

OBJECTIVE: To determine the contents of berberine in Coptis chinensis of Lichuan and establish its best cultivation scheme. METHOD: We used high performance liquid chromatography (HPLC) to measure the contents of berberine at different altitude, growth age, and leaves. RESULTS: Analytic data showed that the growth age and firry woods shading did not affect the contents of berberine in rhizome of Coptis chinensis. Low altitude was more suitable for Coptis chinensis to synthesize berberine. The contents of berberine in rhizome of Coptis chinensis with floral leaf were higher than those with lusterless and lustrous leaves, but no significant difference. CONCLUSION: The synthesis of berberine is closely correlated with shading conditions. It is recommended that the shading ratio should be reduced or the sheds removed in the middle growth age (2-3 years), and then the herb should be reshaped at the last year to enhance the synthesis of berberine, so as to obtain high-quality Coptis chinensis in the harvest.

[Studies on the pharmacological effects of saffron(Crocus sativus L.)--a review].

OBJECTIVE: Reviewing the studies on the chemical components and medicinal value. METHOD: Philological method. RESULT: Saffron is a conventional effective medicine in improving blood circulation and curing the bruise. The late of evidesnces indicate that saffron possesses anticancer activity against a wide spectrum of tumors, such as leukemia, ovarian carcinoma, colon adenocarcinoma, rhabdomyosarcoma, papilloma, squamous cell carcinoma, and soft tissue sarcoma. It has low biochemical toxic effects on animals. In addition, saffron can be used to cure coronary heart disease and hepatitis, and to promote immunity. CONCLUSION: Saffron is a highly valuable medicine. And producing it in a large quantity has a wide application prosperity.