Matrine Mediated Immune Protection in MS by Regulating Gut Microbiota and Production of SCFAs.

There is clearly an unmet need for more effective and safer treatments for multiple sclerosis (MS). Our previous studies showed a significant therapeutic effect of matrine, a monomer of traditional herbal medicine, on experimental autoimmune encephalomyelitis (EAE) mice. To explore the mechanism of matrine action, we used 16S rRNA sequencing technology to determine the gut microbes in matrine-treated EAE mice and controls. The concentrations of short-chain fatty acids (SCFAs) were then tested by metabonomics. Finally, we established pseudo-sterile mice and transplanted into them fecal microbiota, which had been obtained from the high-dose matrine-treated EAE mice to test the effects of matrine. The results showed that matrine could restore the diversity of gut microbiota and promote the production of SCFAs in EAE mice. Transplantation of fecal microbiota from matrine-treated mice significantly alleviated EAE severity, reduced CNS inflammatory infiltration and demyelination, and decreased the level of IL-17 but increased IL-10 in sera of mice. In conclusion, matrine treatment can regulate gut microbiota and metabolites and halt the progression of MS.

Molecular and proteomic response of Pseudomonas fluorescens biofilm cultured on lettuce (Lactuca sativa L.) after ultrasound treatment at different intensity levels.

Ultrasonic treatment is widely used for surface cleaning of vegetables in the processing of agricultural products. In the present study, the molecular and proteomic response of Pseudomonas fluorescens biofilm cultured on lettuce was investigated after ultrasound treatment at different intensity levels. The results show that the biofilm was efficiently removed after ultrasound treatment with intensity higher than 21.06 W/cm2. However, at an intensity of less than 18.42 W/cm2, P. fluorescens was stimulated by ultrasound leading to promoted bacterial growth, extracellular protease activity, extracellular polysaccharide secretion (EPS), and synthesis of acyl-homoserine lactones (AHLs) as quorum-sensing signaling molecules. The expression of biofilm-related genes, stress response, and dual quorum sensing system was upregulated during post-treatment ultrasound. Proteomic analysis showed that ultrasound activated proteins in the flagellar system, which led to changes in bacterial tendency; meanwhile, a large number of proteins in the dual-component system began to be regulated. ABC transporters accelerated the membrane transport of substances inside and outside the cell membrane and equalized the permeability conditions of the cell membrane. In addition, the expression of proteins related to DNA repair was upregulated, suggesting that bacteria repair damaged DNA after ultrasound exposure.

Effect of Kangfuxiaomi suppository on pelvic inflammatory disease in rats.

Pelvic inflammatory disease (PID) is commonly encountered in gynecological practice. Kangfuxiaomi suppository, made from the compound extract of Periplaneta Americana, is a Traditional Chinese Medicine remedy widely used for the treatment of gynecological disorders. This study aimed to preliminarily explore the therapeutic effect of Kangfuxiaomi suppository in a rat model of PID established by chemical injury and pathogen infection. The key parameters assessed were vulvar inflammation score, vaginal + uterine organ index, and serum levels of interleukin (IL)- 8; tumor necrosis factor (TNF)-α; C-reactive protein (CRP); superoxide dismutase (SOD); and malondialdehyde (MDA). In addition, levels of IL-6, cyclooxygenase (COX)- 2, and IL-2 in cervical tissues as well as that of IL-1ß and prostaglandin E-2 (PGE2) in uterine tissues were measured. The expression levels of nuclear factor-kappa B (NF-κB) p65 and Toll-like receptor 4 (TLR4) in uterine tissues were detected by immunohistochemical method. After Kangfuxiaomi suppository treatment, the vulva inflammation score and histopathological score of PID rats showed a tendency to decrease. Serum IL-8, TNF-α, CRP, and MDA levels were reduced, while SOD levels were significantly increased. Levels of IL-6, IL-2, and COX-2 in cervical tissues were somewhat decreased, and PGE2 and IL-1ß levels in uterine tissue were significantly decreased. Moreover, the levels of NF-κB p65 and TLR4 protein expression were also decreased. These findings demonstrated the therapeutic effect of Kangfuxiaomi suppository in PID rats. The underlying mechanism may involve enhanced antioxidant capacity and decreased secretion of proinflammatory factors via the NF-κB/TLR4 signaling pathway.

Bavachin induces apoptosis in colorectal cancer cells through Gadd45a via the MAPK signaling pathway.

Bavachin is a dihydroflavonoid compound isolated from Psoralea corylifolia, and exhibits anti-bacterial, anti-inflammatory, anti-tumor and lipid-lowering activities. Recent attention has gradually drawn on bavachin-induced apoptosis in many human cancer cell lines. However, the anti-cancer effects and related mechanisms in colorectal cancer remain unknown. Here, we investigated the effects of bavachin on colorectal cancer in vivo and in vitro. The results showed that bavachin inhibited the proliferation of human colorectal cancer cells and induce apoptosis. These changes were mediated by activating the MAPK signaling pathway, which significantly up-regulated the expression of Gadd45a. Furthermore, Gadd45a silencing obviously attenuated bavachin-mediated cell apoptosis. Inhibition of the MAPK signaling pathway by JNK/ERK/p38 inhibitors also weakened the up-regulation of Gadd45a by bavachin. The anticancer effect of bavachin was also validated using a mouse xenograft model of human colorectal cancer. In conclusion, these findings suggest that bavachin induces the apoptosis of colorectal cancer cells through activating the MAPK signaling pathway.

Ten new prenylated flavonoids from Macaranga denticulata and their antitumor activities.

Ten new prenylated flavonoids, named denticulains A-J (1-10), together with seven known prenylated flavonoids (11-17) were isolated from Macaranga denticulata. Their structures were elucidated on the basis of detailed spectroscopic analysis and by comparison with literature data. In addition, compounds 1 and 14 inhibited the proliferation of SW620 and HCT-116 cell lines with an IC50 value of 46.08 µM and 56.83 µM, respectively.

Matrine treatment reduces retinal ganglion cell apoptosis in experimental optic neuritis.

Inflammatory demyelination and axonal injury of the optic nerve are hallmarks of optic neuritis (ON), which often occurs in multiple sclerosis and is a major cause of visual disturbance in young adults. Although a high dose of corticosteroids can promote visual recovery, it cannot prevent permanent neuronal damage. Novel and effective therapies are thus required. Given the recently defined capacity of matrine (MAT), a quinolizidine alkaloid derived from the herb Radix Sophorae flavescens, in immunomodulation and neuroprotection, we tested in this study the effect of matrine on rats with experimental autoimmune encephalomyelitis, an animal model of multiple sclerosis. MAT administration, started at disease onset, significantly suppressed optic nerve infiltration and demyelination, with reduced numbers of Iba1+ macrophages/microglia and CD4+ T cells, compared to those from vehicle-treated rats. Increased expression of neurofilaments, an axon marker, reduced numbers of apoptosis in retinal ganglion cells (RGCs). Moreover, MAT treatment promoted Akt phosphorylation and shifted the Bcl-2/Bax ratio back towards an antiapoptotic one, which could be a mechanism for its therapeutic effect in the ON model. Taken as a whole, our results demonstrate that MAT attenuated inflammation, demyelination and axonal loss in the optic nerve, and protected RGCs from inflammation-induced cell death. MAT may therefore have potential as a novel treatment for this disease that may result in blindness.

Illumination of Molecular Pathways in Multiple Sclerosis Lesions and the Immune Mechanism of Matrine Treatment in EAE, a Mouse Model of MS.

The etiology of multiple sclerosis (MS) is not clear, and the treatment of MS presents a great challenge. This study aimed to investigate the pathogenesis and potential therapeutic targets of MS and to define target genes of matrine, a quinolizidine alkaloid component derived from the root of Sophorae flavescens that effectively suppressed experimental autoimmune encephalomyelitis (EAE), an animal model of MS. To this end, the GSE108000 gene data set in the Gene Expression Omnibus Database, which included 7 chronic active MS lesions and 10 control samples of white matter, was analyzed for differentially expressed genes (DEGs). X cell was used to analyze the microenvironmental differences in brain tissue samples of MS patients, including 64 types of immune cells and stromal cells. The biological functions and enriched signaling pathways of DEGs were analyzed by multiple approaches, including GO, KEGG, GSEA, and GSVA. The results by X cell showed significantly increased numbers of immune cell populations in the MS lesions, with decreased erythrocytes, megakaryocytes, adipocytes, keratinocytes, endothelial cells, Th1 cells and Tregs. In GSE108000, there were 637 DEGs, including 428 up-regulated and 209 down-regulated genes. Potential target genes of matrine were then predicted by the network pharmacology method of Traditional Chinese medicine, and 12 key genes were obtained by cross analysis of the target genes of matrine and DEGs in MS lesions. Finally, we confirmed by RT-PCR the predicted expression of these genes in brain tissues of matrine-treated EAE mice. Among these genes, 2 were significantly downregulated and 6 upregulated by matrine treatment, and the significance of this gene regulation was further investigated. In conclusion, our study defined several possible matrine target genes, which can be further elucidated as mechanism(s) of matrine action, and novel targets in the treatment of MS.

Multi-scale Modeling of Nutrient Pollution in the Rivers of China.

Chinese surface waters are severely polluted by nutrients. This study addresses three challenges in nutrient modeling for rivers in China: (1) difficulties in transferring modeling results across biophysical and administrative scales, (2) poor representation of the locations of point sources, and (3) limited incorporation of the direct discharge of manure to rivers. The objective of this study is, therefore, to quantify inputs of nitrogen (N) and phosphorus (P) to Chinese rivers from different sources at multiple scales. We developed a novel multi-scale modeling approach including a detailed, state-of-the-art representation of point sources of nutrients in rivers. The model results show that the river pollution and source attributions differ among spatial scales. Point sources accounted for 75% of the total dissolved phosphorus (TDP) inputs to rivers in China in 2012, and diffuse sources accounted for 72% of the total dissolved nitrogen (TDN) inputs. One-third of the sub-basins accounted for more than half of the pollution. Downscaling to the smallest scale (polygons) reveals that 14% and 9% of the area contribute to more than half of the calculated TDN and TDP pollution, respectively. Sources of pollution vary considerably among and within counties. Clearly, multi-scale modeling may help to develop effective policies for water pollution.

Excess nutrient loads to Lake Taihu: Opportunities for nutrient reduction.

Intensive agriculture and rapid urbanization have increased nutrient inputs to Lake Taihu in recent decades. This resulted in eutrophication. We aim to better understand the sources of river export of total dissolved nitrogen (TDN) and phosphorus (TDP) to Lake Taihu in relation to critical nutrient loads. We implemented the MARINA-Lake (Model to Assess River Inputs of Nutrients to seAs) model for Lake Taihu. The MARINA-Lake model quantifies river export of dissolved inorganic and organic N and P to the lake by source from sub-basins. Results from the PCLake model are used to identify to what extent river export of nutrients exceeds critical loads. We calculate that rivers exported 61 kton of TDN and 2 kton of TDP to Lake Taihu in 2012. More than half of these nutrients were from human activities (e.g., agriculture, urbanization) in Sub-basins I (north) and IV (south). Most of the nutrients were in dissolved inorganic forms. Diffuse sources contributed 90% to river export of TDN with a relatively large share of synthetic fertilizers. Point sources contributed 52% to river export of TDP with a relatively large share of sewage systems. The relative shares of diffuse and point sources varied greatly among nutrient forms and sub-basins. To meet critical loads, river export of TDN and TDP needs to be reduced by 46-92%, depending on the desired level of chlorophyll-a. There are different opportunities to meet the critical loads. Reducing N inputs from synthetic fertilizers and P from sewage systems may be sufficient to meet the least strict critical loads. A combination of reductions in diffuse and point sources is needed to meet the most strict critical loads. Combining improved nutrient use efficiencies and best available technologies in wastewater treatment may be an effective opportunity. Our study can support the formulation of effective solutions for lake restoration.

GC-MS Study of the Chemical Components of Different Aquilaria sinensis (Lour.) Gilgorgans and Agarwood from Different Asian Countries.

As a traditional medicinal herb and valuable natural spice in China, Aquilaria sinensis (Lour.) Gilg has many significant pharmacological effects. Agarwood is the resinous heartwood acquired from wounded A. sinensis trees, and is widely used in pharmaceuticals owing to its excellent medicinal value. In this study, the chemical composition of volatile components and alcohol extracts from different organs of A. sinensis and agarwoods grown in different regions were investigated using GC-MS. The results showed that Vietnam agarwood had the highest moisture content, which was attributed to the local climate, while the fruit and bark of A. sinensis had higher moisture contents than the other organs. The volatile components of A. sinensis organs included 3-ethyl-5-(2-ethylbutyl)-octadecane, oleic acid 3-(octadecyloxy) propyl ester, and docosanoic acid 1,2,3-propanetriyl ester, while the alcohol extracts of A. sinensis organs contained benzoic acid ethyl ester, hexadecanoic acid ethyl ester, oleic acid, and n-hexadecanoic acid. Furthermore, the main active ingredients in agarwood from different habitats were sesquiterpenoids, aromatic species, and chromone compounds. The role of chromone compound 2-phenylethyl-benzopyran as an elicitor and the mechanism of agarwood formation were also investigated. Antioxidant tests showed that essential oils from agarwood and A. sinensis had antioxidant capacities by comparison with butylated hydroxytoluene and vitamin E. An antibacterial activity test showed that the inhibition effect of the essential oil was better against Gram-positive bacteria than against Gram-negative bacteria.

Hotspots for Nitrogen and Phosphorus Losses from Food Production in China: A County-Scale Analysis.

Food production in China results in large losses of nitrogen (N) and phosphorus (P) to the environment. Our objective is to identify hotspots for N and P losses to the environment from food production in China at the county scale. To do this, we used the NUFER (Nutrient flows in Food chains, Environment and Resources use) model. Between 1990 and 2012, the hotspot area expanded by a factor of 3 for N, and 24 for P. In 2012 most hotspots were found in the North China Plain. Hotspots covered less than 10% of the Chinese land area, but contributed by more than half to N and P losses to the environment. Direct discharge of animal manure to rivers was an important cause of N and P losses. Food production was found to be more intensive in hotspots than in other counties. Synthetic fertilizer use and animal numbers in hotspots were a factor of 4-5 higher than in other counties in 2012. Also the number of people working in food production and the incomes of farmers are higher in hotspots than in other counties. This study concludes with suggestions for region-specific pollution control technologies for food production in China.

Exocyst subunit SEC3A marks the germination site and is essential for pollen germination in Arabidopsis thaliana.

Arabidopsis exocyst subunit SEC3A has been reported to participate in embryo development. Here we report that SEC3A is involved during pollen germination. A T-DNA insertion in SEC3A leads to an absolute, male-specific transmission defect that can be complemented by the expression of SEC3A coding sequence from the LAT52 promoter or SEC3A genomic DNA. No obvious abnormalities in the microgametogenesis are observed in the sec3a/SEC3A mutant, however, in vitro and in vivo pollen germination are defective. Further studies reveal that the callose, pectin, and cellulose are apparently not deposited at the germination site during pollen germination. SEC3A is expressed ubiquitously, including in pollen grains and pollen tubes. Notably, SEC3A-GFP fusion proteins are specifically recruited to the future pollen germination site. This particular localization pattern is independent of phosphatidylinositol 4,5-bisphosphate (PI-4,5P2), although SEC3-HIS fusion proteins are able to bind to several phosphoinositols in vitro. These results suggest that SEC3A plays an important role in the establishment of the polar site for pollen germination.

The MARINA model (Model to Assess River Inputs of Nutrients to seAs): Model description and results for China.

Chinese agriculture has been developing fast towards industrial food production systems that discharge nutrient-rich wastewater into rivers. As a result, nutrient export by rivers has been increasing, resulting in coastal water pollution. We developed a Model to Assess River Inputs of Nutrients to seAs (MARINA) for China. The MARINA Nutrient Model quantifies river export of nutrients by source at the sub-basin scale as a function of human activities on land. MARINA is a downscaled version for China of the Global NEWS-2 (Nutrient Export from WaterSheds) model with an improved approach for nutrient losses from animal production and population. We use the model to quantify dissolved inorganic and organic nitrogen (N) and phosphorus (P) export by six large rivers draining into the Bohai Gulf (Yellow, Hai, Liao), Yellow Sea (Yangtze, Huai) and South China Sea (Pearl) in 1970, 2000 and 2050. We addressed uncertainties in the MARINA Nutrient model. Between 1970 and 2000 river export of dissolved N and P increased by a factor of 2-8 depending on sea and nutrient form. Thus, the risk for coastal eutrophication increased. Direct losses of manure to rivers contribute to 60-78% of nutrient inputs to the Bohai Gulf and 20-74% of nutrient inputs to the other seas in 2000. Sewage is an important source of dissolved inorganic P, and synthetic fertilizers of dissolved inorganic N. Over half of the nutrients exported by the Yangtze and Pearl rivers originated from human activities in downstream and middlestream sub-basins. The Yellow River exported up to 70% of dissolved inorganic N and P from downstream sub-basins and of dissolved organic N and P from middlestream sub-basins. Rivers draining into the Bohai Gulf are drier, and thus transport fewer nutrients. For the future we calculate further increases in river export of nutrients. The MARINA Nutrient model quantifies the main sources of coastal water pollution for sub-basins. This information can contribute to formulation of effective management options to reduce nutrient pollution of Chinese seas in the future.

Chemomic and chemometric approach based on ultra-fast liquid chromatography with ion trap time-of-flight mass spectrometry to reveal the difference in the chemical composition between Da-Cheng-Qi decoction and its three constitutional herbal medicines.

Da-Cheng-Qi decoction (DCQT) is a traditional purgative Chinese decoction with a history of 2000 years. To study the effect of interactions between the ingredients on the overall chemical composition of DCQT, a chemomic and chemometric approach based on ultra-fast liquid chromatography with ion trap time-of-flight mass spectrometry was developed and validated. After mixing and decocting all four ingredients to make the DCQT, the concentrations of some chemicals are significantly different from those in single herb decoction and 24 of them were identified and tentatively characterized by comparing their data with those of standard compounds or literature data. No new chemicals were formed during mixing and decoction. Our findings indicated that there are interactions between these natural medicines during the mixing and preparation process. The 24 identified chemicals could be used as chemical markers for optimizing prescription and evaluation of consistent quality, and the strategy in the present study could be applied for other multiherb formulae.