Epigallocatechin gallate suppresses mitotic clonal expansion and adipogenic differentiation of preadipocytes through impeding JAK2/STAT3-mediated transcriptional cascades.

BACKGROUND: Mitotic clonal expansion (MCE) is a prerequisite for preadipocyte differentiation and adipogenesis. Epigallocatechin gallate (EGCG) has been shown to inhibit preadipocyte differentiation. However, the exact molecular mechanisms are still elusive. PURPOSE: This study investigated whether EGCG could inhibit adipogenesis and lipid accumulation by regulating the cell cycle in the MCE phase of adipogenesis and its underlying molecular mechanisms. METHOD: 3T3-L1 preadipocytes were induced to differentiate by a differentiation cocktail (DMI) and were treated with EGCG (25-100 µM) for 9, 18, and 24 h to examine the effect on MCE, or eight days to examine the effect on terminal differentiation. C57BL/6 mice were fed a high-fat diet (HFD) for three months to induce obesity and were given EGCG (50 or 100 mg/kg) daily by gavage. RESULTS: We showed that EGCG significantly inhibited terminal adipogenesis and lipid accumulation in 3T3-L1 cells and decreased expressions of PPARγ, C/EBPα, and FASN. Notably, at the MCE phase, EGCG regulated the cell cycle in sequential order, induced G0/G1 arrest at 18 h, and inhibited the G2/M phase at 24 h upon DMI treatment. Meanwhile, EGCG regulated the expressions of cell cycle regulators (cyclin D1, cyclin E1, CDK4, CDK6, cyclin B1, cyclin B2, p16, and p27), and decreased C/EBPß, PPARγ, and C/EBPα expressions at MCE. Mechanistic studies using STAT3 agonist Colivelin and antagonist C188-9 revealed that EGCG-induced cell cycle arrest in the MCE phase and terminal adipocyte differentiation was mediated by the inhibition of JAK2/STAT3 signaling cascades and STAT3 (Tyr705) nuclear translocation. Furthermore, EGCG significantly protected mice from HFD-induced obesity, reduced body weight and lipid accumulations in adipose tissues, reduced hyperlipidemia and leptin levels, and improved glucose intolerance and insulin sensitivity. Moreover, RNA sequencing (RNA-seq) analysis showed that the cell cycle changes in epididymal white adipose tissue (eWAT) were significantly enriched upon EGCG treatment. We further verified that EGCG treatment significantly reduced expressions of adipogenic factors, cell cycle regulators, and p-STAT3 in eWAT. CONCLUSION: EGCG inhibits MCE, resulting in the inhibition of early and terminal adipocyte differentiation and lipid accumulation, which were mediated by inhibiting p-STAT3 nucleus translocation and activation.

Diversity-oriented synthesis of diterpenoid alkaloids yields a potent anti-inflammatory agent.

BACKGROUND: The diterpenoid alkaloids belong to a highly esteemed group of natural compounds, which display significant biological activities. It is a productive strategy to expand the chemical space of these intriguing natural compounds for drug discovery. METHODS: We prepared a series of new derivatives bearing diverse skeletons and functionalities from the diterpenoid alkaloids deltaline and talatisamine based on a diversity-oriented synthesis strategy. The anti-inflammatory activity of these derivatives was initially screened and evaluated by the release of nitric oxide (NO), tumor necrosis factor (TNF-α), and interleukin-6 (IL-6) in lipopolysaccharide (LPS)-activated RAW264.7 cells. Futhermore, the anti-inflammatory activity of the representative derivative 31a was validated in various inflammatory animal models, including phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced mice ear edema, LPS-stimulated acute kidney injury, and collagen-induced arthritis (CIA). RESULTS: It was found that several derivatives were able to suppress the secretion of NO, TNF-α, and IL-6 in LPS-activated RAW264.7 cells. Compound 31a, one of the representative derivatives named as deltanaline, demonstrated the strongest anti-inflammatory effects in LPS-activated macrophages and three different animal models of inflammatory diseases by inhibiting nuclear factor kappa-B (NF-κB)/mitogen-activated protein kinase (MAPK) signaling and inducing autophagy. CONCLUSION: Deltanaline is a new structural compound derived from natural diterpenoid alkaloids, which may serve as a new lead compound for the treatment of inflammatory diseases.

Ganoderma lucidum polysaccharide inhibits HSC activation and liver fibrosis via targeting inflammation, apoptosis, cell cycle, and ECM-receptor interaction mediated by TGF-ß/Smad signaling.

BACKGROUND: Ganoderma lucidum polysaccharide (GLP) has many biological properties, however, the anti-fibrosis effect of GLP is unknown at present. PURPOSE: This study aimed to examine the anti-fibrogenic effect of GLP and its underlying molecular mechanisms in vivo and in vitro. STUDY DESIGN: Both CCl4-induced mouse and TGF-ß1-induced HSC-T6 cellular models of fibrosis were established to examine the anti-fibrogenic effect of a water-soluble GLP (25 kDa) extracted from the sporoderm-removed spores of G. lucidum.. METHOD: Serum markers of liver injury, histology and fibrosis of liver tissues, and collagen formation were examined using an automatic biochemical analyzer, H&E staining, Sirius red staining, immunohistochemistry, immunofluorescence, ELISA, Western blotting, and qRT-PCR. RNA-sequencing, enrichment pathway analysis, Western blotting, qRT-PCR, and flow cytometry were employed to identify the potential molecular targets and signaling pathways that are responsible for the anti-fibrotic effect of GLP. RESULTS: We showed that GLP (150 and 300 mg/kg) significantly inhibited hepatic fibrogenesis and inflammation in CCl4-treated mice as mediated by the TLR4/NF-κB/MyD88 signaling pathway. We further demonstrated that GLP significantly inhibited hepatic stellate cell (HSCs) activation in mice and in TGF-ß1-induced HSC-T6 cells as manifested by reduced collagen I and a-SMA expressions. RNA-sequencing uncovered inflammation, apoptosis, cell cycle, ECM-receptor interaction, TLR4/NF-κB, and TGF-ß/Smad signalings as major pathways suppressed by GLP administration. Further studies demonstrated that GLP elicits anti-fibrotic actions that are associated with a novel dual effect on apoptosis in vivo (inhibit) or in vitro (promote), suppression of cell cycle in vivo, induction of S phase arrest in vitro, and attenuation of ECM-receptor interaction-associated molecule expressions including integrins ITGA6 and ITGA8. Furthermore, GLP significantly inhibited the TGF-ß/Smad signaling in mice, and reduced TGF-ß1 or its agonist SRI-011381-induced Smad2 and Smad3 phosphorylations, but increased Samd7 expression in HSC-T6 cells. CONCLUSION: This study provides the first evidence that GLP could be a promising dietary strategy for treating liver fibrosis, which protects against liver fibrosis and HSC activation through targeting inflammation, apoptosis, cell cycle, and ECM-receptor interactions that are mediated by TGF-ß/Smad signaling.

Integrating UHPLC-MS/MS quantitative analysis and exogenous purine supplementation to elucidate the antidepressant mechanism of Chaigui granules by regulating purine metabolism.

Chaigui granules (CG) are a compound composed of six herbal medicines with significant antidepressant effects. However, the antidepressant mechanism of CG remains unclear. In the present study, we attempted to elucidate the antidepressant mechanism of CG by regulating purine metabolism and purinergic signaling. First, the regulatory effect of CG on purine metabolites in the prefrontal cortex (PFC) of chronic unpredictable mild stress (CUMS) rats was analyzed by ultra high-performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) targeted quantitative analysis. Meanwhile, purinergic receptors (P2X7 receptor (P2X7R), A1 receptor (A1R) and A2A receptor (A2AR)) and signaling pathways (nod-like receptor protein 3 (NLRP3) inflammasome pathway and cyclic adenosine monophosphate (cAMP)-protein kinase A (PKA) pathway) associated with purine metabolism were analyzed by western blotting and enzyme-linked immunosorbent assay (ELISA). Besides, antidepressant mechanism of CG by modulating purine metabolites to activate purinergic receptors and related signaling pathways was dissected by exogenous supplementation of purine metabolites and antagonism of purinergic receptors in vitro. An in vivo study showed that the decrease in xanthine and the increase in four purine nucleosides were closely related to the antidepressant effects of CG. Additionally, purinergic receptors (P2X7R, A1R and A2AR) and related signaling pathways (NLRP3 inflammasome pathway and cAMP-PKA pathway) were also significantly regulated by CG. The results of exogenous supplementation of purine metabolites and antagonism of purinergic receptors showed that excessive accumulation of xanthine led to activation of the P2X7R-NLRP3 inflammasome pathway, and the reduction of adenosine and inosine inhibited the A1R-cAMP-PKA pathway, which was significantly ameliorated by CG. Overall, CG could promote neuroprotection and ultimately play an antidepressant role by inhibiting the xanthine-P2X7R-NLRP3 inflammasome pathway and activating the adenosine/inosine-A1R-cAMP-PKA pathway.

The Effects of Adding Di-Tan Decoction (DTD) and/or Electroacupuncture (EA) to Standard Swallowing Rehabilitation Training (SRT) for Improving Poststroke Dysphagia (PSD): A Pilot, Single-Centred, Randomized Trial.

Objectives: To evaluate the effect of adding Di-tan decoction (DTD) and/or electroacupuncture (EA) to standard swallowing rehabilitation training (SRT) on improving PSD. Methods: In total, 80 PSD patients were enrolled and randomly assigned to the DTD, EA, DTD + EA or control group at a 1 : 1 : 1 : 1 ratio. All patients received basic treatment and standard SRT. The DTD group received DTD orally, the EA group received EA, the DTD + EA group received both DTD and EA simultaneously, and the control group received only basic treatment and standard SRT. The interventions lasted for 4 weeks. The outcome measurements included the Standardized Swallowing Assessment (SSA) and Swallowing-Quality of Life (SWAL-QOL), performed and scored from baseline to 2, 4, and 6 weeks after intervention, and the Videofluoroscopic Dysphagia Scale (VDS), scored at baseline and 4 weeks after intervention. Scores were compared over time by repeated-measures analysis of variance (ANOVA) among all groups. Interactions between interventions were explored using factorial design analysis. Results: (1) The effective rates (ERs) for PSD treatment were higher in the DTD, EA and DTD + EA groups than in the control group (all P < 0.05). The ER was higher in the DTD + EA group than in the DTD or EA group (both P < 0.05). (2) There were significant group effects, time effects and interactions for the SSA and SWAL-QOL scores (all P < 0.05). All groups showed decreasing trends in SSA scores and increasing trends in SWAL-QOL scores over time from baseline to 6 weeks after intervention (all P < 0.01). (3) Factorial design analysis for ΔVDS showed that there was a significant main effect for DTD intervention (F = 11.877, P < 0.01) and for EA intervention (F = 29.357, P < 0.01). However, there was no significant interaction effect between DTD and EA (F = 0.133, P = 0.717). Multiple comparisons showed that the DTD, EA and DTD + EA groups all had higher ΔVDS values than the control group (P < 0.05). The DTD + EA group had a higher ΔVDS than the DTD or EA group (both P < 0.05). (4) Most adverse reactions were mild and transient. Conclusions: Adding DTD or EA to SRT can better improve PSD than applying SRT alone. Adding DTD and EA simultaneously can accelerate and amplify the recovery of swallowing function versus DTD or EA alone, and both are effective and safe treatments, alone or jointly, for PSD and are a powerful supplement to routine treatments.

Exploration of the role of bound polyphenols on tea residues dietary fiber improving diabetic hepatorenal injury and metabolic disorders.

Consumption of tea residues dietary fiber (TRDF) contributed to the relief of hyperglycemia symptoms in type 2 diabetes (T2D). Given the properties of TRDF abundant in bound polyphenols, the research intended to evaluate the effect of the presence or absence of bound polyphenols in TRDF on the improvement of diabetic complications (liver and kidney injury, metabolic disorders) in T2D rats induced by high-fat diet and streptozocin injection. Our results revealed that the presence of bound polyphenols in TRDF was remarkably beneficial for the amelioration of liver and kidney damage caused by T2D, which was supported by significant differences in activities of serum glutamic oxaloacetic transaminase (AST) and glutamic-pyruvic transaminase (ALT), contents of inflammatory factors in liver and kidney, the levels of kidney oxidative stress, as well as histopathological status between TRDF and bound polyphenols removed-TRDF (TRDF-DF) groups. In addition, metabolomic analysis revealed that TRDF interventions could increase the levels of metabolites such as S-Adenosylmethionine, L-Homophenylalanine and Riboflavin, as well as differ in the regulation of the metabolic pathways including arachidonic acid metabolism and cysteine and methionine metabolism as compared to TRDF-DF without bound polyphenols. These results suggested that bound polyphenols ensured the health-promoting effects for T2D complications of TRDF.

Metabonomics combined with 16S rRNA sequencing to elucidate the hypoglycemic effect of dietary fiber from tea residues.

Tea residues are rich in dietary fiber, which possesses excellent physicochemical and functional properties in vitro. However, the hypoglycemic effect and mechanism of dietary fiber from tea residues are not clear. The study aimed to investigate the potential hypoglycemic effect of dietary fiber obtained from tea residues fermentation (TRDF) and reveal its related mechanisms of action in terms of both intestinal flora and metabolomics. The type 2 diabetes (T2D) rat model induced by high-fat diet and streptozotocin injection was applied in this study. Four weeks of TRDF intervention could remarkably ameliorate hyperglycemia, severe oxidative stress and insulin resistance of diabetic rats. Additionally, there was a significant increase of short chain fatty acids (SCFAs) concentrations in feces of diabetic rats after TRDF intervention. Furthermore, TRDF played a positive role in relieving intestinal microbiota dysbiosis by enriching beneficial bacteria (S24-7 and Prevotellaceae) and inhibiting harmful bacteria (Desulfovibrionaceae and Clostridiaceae). Metabolomic analysis showed that TRDF improved the amino acid metabolism and citrate cycle. The study elaborated on the hypoglycemic effect and potential mechanisms of TRDF through multiple pathways of gut microbiota and metabolites, which could provide theoretical basis for TRDF as a dietary supplement to manage T2D.

Elucidation of the interaction effect between dietary fiber and bound polyphenol components on the anti-hyperglycemic activity of tea residue dietary fiber.

Dietary fiber intake is beneficial for the prevention of some chronic metabolic diseases. Considering the characteristic that dietary fiber from tea residues (TRDF) is rich in bound polyphenols, the study aimed to elucidate the interaction effect between dietary fiber components (TRDF-DF) and bound polyphenol components (TRDF-BP) on the anti-hyperglycemic activity of TRDF. A type 2 diabetes (T2D) rat model induced by high-fat diet and streptozotocin injection was applied in this study. The results showed that bound polyphenol components rather than dietary fiber components were essential for the anti-hyperglycemic activity of TRDF, as evidenced by remarkable differences in fasting blood glucose (FBG), the insulin resistance index (HOMA-IR) and the levels of serum oxidative stress between the TRDF and TRDF-DF groups, as well as the up-regulation of the expression of insulin signaling pathway-related proteins in the liver after TRDF and TRDF-BP administration. In addition, the synergistic effect between TRDF-BP and TRDF-DF components modulated gut microbiota dysbiosis and increased the content of short chain fatty acids (SCFAs) via enriching beneficial bacteria and inhibiting harmful bacteria. The role of TRDF-BP and TRDF-DF as well as their interaction effect on the anti-hyperglycemic activity of TRDF are elucidated, which can provide theoretical basis for TRDF as a dietary supplement to manage T2D.

Network Pharmacology Analysis on Zhichan Powder in the Treatment of Parkinson's Disease.

AIMS AND OBJECTIVE: Effective components and the mechanism of action of Zhichan powder for the treatment of Parkinson's disease were researched at a systematic level. MATERIALS AND METHODS: Screening of active components in Zhichan powder for the treatment of Parkinson's disease was conducted using the Traditional Chinese Medicine Systems Pharmacology database, and a medicine-target-disease network was established with computational network pharmacology. RESULTS: By using network pharmacology methods, we identified 18 major active components in Zhichan powder through screening, indicating a connection between chemical components of this Traditional Chinese Medicine and Parkinson's disease-related targets. CONCLUSION: The medicine-target-disease system of Zhichan powder established by network pharmacology permitted visualization of clustering and differences among chemical components in this prescription, as well as the complex mechanism of molecular activities among those effective components, relevant targets, pathways, and the disease. Thus, our results provide a new perspective and method for revealing the mechanism of action of Traditional Chinese Medicine prescriptions.

Effect of Xiaoaiping injection on advanced hepatocellular carcinoma in patients.

OBJECTIVE: To investigate the effect of Xiaoaiping Injection (XAP) on advanced hepatocellular carcinoma (HCC) in patients. METHODS: Sixty-eight patients with advanced HCC were assigned to a control group of 36 and a treatment group of 32. The control group was treated with best supportive treatment (BST) and the treatment group was given XAP plus BST. XAP was administered daily by i.v. and the treatment course was lasted for 30 days for both groups. The immediate therapeutic efficacy, Karnofsky performance status (KPS) scores, and the changes in immunity indicators (CD3+, CD4+ and CD8+ T cells) were measured and compared before and after treatment. The progression-free survival (PFS) rate and overall survival (OS) rate in the 2 groups were analyzed. RESULTS: The immediate therapeutic efficacy and KPS of the treatment group were better than those in the control (P < 0.05). Patients in the treatment group had higher percentages of CD3 and CD4 T-lymphocytes in peripheral blood than those in the control group (P < 0.05). The median survival time was 27.0 weeks in the treatment group and 24.5 weeks in the control group. The 6-months cumulative survival rates in the treatment and control groups were 33.3% and 25.0%, respectively, with no significant difference (P > 0.05). The PFS was 18 weeks in the treatment group and 15 weeks in control group (P < 0.05). CONCLUSION: XAP enhances the quality of life (QOL) of patients with advanced HCC, improves their immunity and extends their PFS.

The mechanism of astragaloside IV promoting sciatic nerve regeneration.

3-O-beta-D-xylopyranosyl-6-O-beta-D-glucopyranosyl-cycloastragenol (astragaloside IV), the main active component of the traditional Chinese medicine astragalus membranaceus, has been shown to be neuroprotective. This study investigated whether astragaloside IV could promote the repair of injured sciatic nerve. Denervated sciatic nerve of mice was subjected to anastomosis. The mice were intraperitoneally injected with 10, 5, 2.5 mg/kg astragaloside IV per day for 8 consecutive days. Western blot assay and real-time PCR results demonstrated that growth-associated protein-43 expression was upregulated in mouse spinal cord segments L4-6 after intervention with 10, 5, 2.5 mg/kg astragaloside IV per day in a dose-dependent manner. Luxol fast blue staining and electrophysiological detection suggested that astragaloside IV elevated the number and diameter of myelinated nerve fibers, and simultaneously increased motor nerve conduction velocity and action potential amplitude in the sciatic nerve of mice. These results indicated that astragaloside IV contributed to sciatic nerve regeneration and functional recovery in mice. The mechanism underlying this effect may be associated with the upregulation of growth-associated protein-43 expression.

Ursolic acid induces neural regeneration after sciatic nerve injury.

In this study, we aimed to explore the role of ursolic acid in the neural regeneration of the injured sciatic nerve. BALB/c mice were used to establish models of sciatic nerve injury through unilateral sciatic nerve complete transection and microscopic anastomosis at 0.5 cm below the ischial tube-rosity. The successfully generated model mice were treated with 10, 5, or 2.5 mg/kg ursolic acid via intraperitoneal injection. Enzyme-linked immunosorbent assay results showed that serum S100 protein expression level gradually increased at 1-4 weeks after sciatic nerve injury, and significantly decreased at 8 weeks. As such, ursolic acid has the capacity to significantly increase S100 protein expression levels. Real-time quantitative PCR showed that S100 mRNA expression in the L4-6 segments on the injury side was increased after ursolic acid treatment. In addition, the muscular mass index in the soleus muscle was also increased in mice treated with ursolic acid. Toluidine blue staining revealed that the quantity and average diameter of myelinated nerve fibers in the injured sciatic nerve were significantly increased after treatment with ursolic acid. 10 and 5 mg/kg of ursolic acid produced stronger effects than 2.5 mg/kg of ursolic acid. Our findings indicate that ursolic acid can dose-dependently increase S100 expression and promote neural regeneration in BALB/c mice following sciatic nerve injury.

Use of acupuncture to treat cerebral infarction in the last 10 years: A Scopus-based literature analysis.

OBJECTIVE: To identify global research trends in the use of acupuncture to treat cerebral infarction. DATA RETRIEVAL: We performed a bibliometric analysis of studies on the use of acupuncture to treat cerebral infarction published during 2002-2011, retrieved from Scopus, using the key words of acupuncture and cerebral infarction or ischemic stroke. INCLUSION CRITERIA: peer-reviewed articles on the use of acupuncture to treat cerebral infarction indexed in Scopus and published between 2002 and 2011; types of publications were original research articles, reviews, meeting abstracts, proceedings papers, book chapters, editorial material, and news items. EXCLUSION CRITERIA: articles that required manual searching or telephone access; documents that were not published in the public domain; and corrected papers. MAIN OUTCOME MEASURES: (a) Annual publication output; (b) language of publication; (c) type of publication; (d) key words of publication; (e) publication by research field; (f) publication by journal; (g) publication by country and institution; (h) publication by author; (i) most-cited papers between 2002 and 2006; and (j) most-cited papers between 2007 and 2011. RESULTS: A total of 160 publications on the use of acupuncture to treat cerebral infarction from 2002-2011 were retrieved from Scopus. The number of publications increased gradually over the 10-year study period; most were written in Chinese or English. Articles and reviews constituted the major types. The most frequent key word used was acupuncture. The most prolific journals in this area were Zhongguo Zhen Jiu and the Chinese Journal of Clinical Rehabilitation. Of the 160 publications retrieved, half came from Chinese authors and institutions. Tianjin University of Traditional Chinese Medicine was the most prolific research institute. Two papers were cited 30 times; they were published in 2002 and 2009, respectively. CONCLUSION: In the field of neuroscience, there is little literature on acupuncture for cerebral infarction. The most-cited papers were cited 30 times in the past 3 years. We believe that, with advances in the study of mechanisms in neurobiology, research on acupuncture will also advance and will become the concern of more scholars.

Zhichan powder regulates nigrostriatal dopamine synthesis and metabolism in Parkinson's disease rats.

In this study, rat models of Parkinson's disease induced by substantia nigra injection of 6-hydroxy-dopamine were intragastrically administered Zhichan powder daily for 50 days. Reverse transcription PCR results showed that tyrosine hydroxylase mRNA expression in the rat substantia nigra was significantly increased, while monoamine oxidase B mRNA expression was significantly decreased in the Zhichan powder group, compared with the model group. In addition, the levels of striatal dopamine and homovanillic acid, the ratio of dopamine to homovanillic acid, and the activity of blood superoxide dismutase were all higher in the Zhichan powder group than in the model group, but the content of malondialdehyde in blood was lower. Our experimental findings indicate that Zhichan powder has an antioxidant effect, it can regulate the expression of monoamine oxidase B and tyrosine hydroxylase in the substantia nigra of Parkinson's disease rats, and it can facilitate the secretion of striatal dopamine and its metabolite homovanillic acid.

Effects of Zhichan powder on signal transduction and apoptosis-associated gene expression in the substantia nigra of Parkinson's disease rats.

Previous studies have shown that Zhichan powder elevated immunity and suppressed oxidation in mice. Rat models of Parkinson's disease were induced by stereotaxically injecting 6-hydroxydopamine into the substantia nigra. The rat models were intragastrically treated with Zhichan powder, which is composed of milkvetch root, ginseng, bunge swallowwort root, himalayan teasel root, Magnolia officinalis, Ligustrum lucidum Ait. and szechwan lovage rhizome. Immunohistochemistry and reverse transcription-PCR results demonstrated that mRNA and protein expression of tumor necrosis factor receptor 1, Fas, caspase-8, cytochrome C, Bax, caspase-3, and p53 significantly increased, but Bcl-2 expression significantly decreased in the substantia nigra of rats with Parkinson's disease. Following Zhichan powder administration, mRNA and protein expression of tumor necrosis factor receptor 1, Fas, caspase-8, cytochrome C, Bax, caspase-3, and p53 diminished, but Bcl-2 expression increased in the rat substantia nigra. These results indicate that Zhichan powder regulates signal transduction protein expression, inhibits apoptosis, and exerts therapeutic effects on Parkinson's disease.

Effect of ω-3 polyunsaturated fatty acid on toll-like receptors in patients with severe multiple trauma.

This study examined the effects of ω-3 polyunsaturated fatty acid (ω-3PUFA) on the expression of toll-like receptor 2 (TLR2), toll-like receptor 4 (TLR4) and some related inflammatory factors in peripheral blood mononuclear cells (PBMCs) of patients with early-stage severe multiple trauma. Thirty-two patients who were admitted to the Department of Traumatic Surgery, Tongji Hospital (Wuhan, China) between May 2010 and November 2010, and diagnosed as having severe multiple trauma with a injury severity score (ISS) no less than 16, were enrolled in the study and divided into two groups at random (n=16 in each): ω-3PUFA group and control group in which routine parenteral nutrition supplemented with ω-3PUFA or not was administered to the patients in two groups for consecutive 7 days. Peripheral blood from these patients was collected within 2 h of admission (day 0), and 1, 3, 5 and 7 days after the nutritional support. PBMCs were isolated and used for detection of the mRNA and protein expression of TLR2 and TLR4 by using real-time PCR and flow cytometry respectively, the levels of NF-κB by quantum dots-based immunofluorescence assay, the levels of TNF-α, IL-2, IL-6 and COX-2 by ELISA, respectively. The results showed that the mRNA and protein expression of TLR2 and TLR4 in PBMCs was significantly lower in ω-3PUFA group than in control group 5 and 7 days after nutrition support (both P<0.05). The levels of TNF-α, IL-2, IL-6 and COX-2 were found to be substantially decreased in PBMCs in ω-3PUFA group as compared with control group at 5th and 7th day (P<0.05 for all). It was concluded that ω-3PUFA can remarkably decrease the expression of TLR2, TLR4 and some related inflammatory factors in NF-κB signaling pathway in PBMCs of patients with severe multiple trauma, which suggests that ω-3PUFA may suppress the excessive inflammatory response meditated by the TLRs/NF-κB signaling pathway.

Experimental analysis of a nitrogen removal process simulation of wastewater land treatment under three different wheat planting densities.

Nitrogen contaminant transport, transformation and uptake simulation experiments were conducted in green house under three different planting density of winter wheat. They were Group A, planting density of 0.0208 plants/cm2, Group B, 0.1042 plants/cm2, and Group C, 0.1415 plants/cm2. The capacity and ratio of nitrogen removal were different on three kinds of conditions of wastewater land treatment. From analysis of wastewater treatment capacity, wastewater concentration and irrigation intensity for Group C were suitable and nitrogen quantity added was 2 times of that for Group B, 2.6 times for Group A while nitrogen residue was only 7.06%. Hence, wastewater irrigation and treatment design with purpose of waste water treatment should select the design with maximum capacity, optimal removal ratio and least residue in soil, which was closely related to crop planting density, crop growth status and also background nitrogen quantity in soil.