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Objectives: Colitis is a global disease usually accompanied by intestinal epithelial damage and intestinal inflammation, and an increasing number of studies have found natural products to be highly effective in treating colitis. Anemoside B4 (AB4), an abundant saponin isolated from Pulsatilla chinensis (Bunge), which was found to have strong anti-inflammatory activity. However, the exact molecular mechanisms and direct targets of AB4 in the treatment of colitis remain to be discovered. Methods: The anti-inflammatory activities of AB4 were verified in LPS-induced cell models and 2, 4, 6-trinitrobenzene sulfonic (TNBS) or dextran sulfate sodium (DSS)-induced colitis mice and rat models. The molecular target of AB4 was identified by affinity chromatography analysis using chemical probes derived from AB4. Experiments including proteomics, molecular docking, biotin pull-down, surface plasmon resonance (SPR), and cellular thermal shift assay (CETSA) were used to confirm the binding of AB4 to its molecular target. Overexpression of pyruvate carboxylase (PC) and PC agonist were used to study the effects of PC on the anti-inflammatory and metabolic regulation of AB4 in vitro and in vivo. Results: AB4 not only significantly inhibited LPS-induced NF-κB activation and increased ROS levels in THP-1 cells, but also suppressed TNBS/DSS-induced colonic inflammation in mice and rats. The molecular target of AB4 was identified as PC, a key enzyme related to fatty acid, amino acid and tricarboxylic acid (TCA) cycle. We next demonstrated that AB4 specifically bound to the His879 site of PC and altered the protein's spatial conformation, thereby affecting the enzymatic activity of PC. LPS activated NF-κB pathway and increased PC activity, which caused metabolic reprogramming, while AB4 reversed this phenomenon by inhibiting the PC activity. In vivo studies showed that diisopropylamine dichloroacetate (DADA), a PC agonist, eliminated the therapeutic effects of AB4 by changing the metabolic rearrangement of intestinal tissues in colitis mice. Conclusion: We identified PC as a direct cellular target of AB4 in the modulation of inflammation, especially colitis. Moreover, PC/pyruvate metabolism/NF-κB is crucial for LPS-driven inflammation and oxidative stress. These findings shed more light on the possibilities of PC as a potential new target for treating colitis. [ABSTRACT FROM AUTHOR]

Background: As the Diagnostic and Statistical Manual of Mental Disorders fifth version (DSM-5) was published, the Kiddie-Schedule for Affective Disorders and Schizophrenia-Present and Lifetime version (K-SADS-PL) was modified to adapt the new version (K-SADS-PL DSM-5). We translated it to Chinese (K-SADS-PL-C DSM-5) and described its reliability and validity.Methods: A total of 154 groups of 6 to 18-year-old children and their guardians were included. Trained interviewers interviewed subjects using the K-SADS-PL-C DSM-5. Interrater reliability was assessed by audio recording. Parent-reported scales, like child behavior checklist (CBCL), the Chinese version of Swan-son Nolan and Pelham, version IV scale-parent form (SNAP-IV), social responsiveness scale (SRS-1), and children-reported scales like depression self-rating scale for children (DSRSC) and the screen for child anxiety related emotional disorders (SCARED) were used to examine the validity of depressive disorder, ADHD, ASD, and ODD.Results: The K-SADS-PL-C DSM-5 had fair to excellent interrater (0.537-1.000) and test-retest (0.468-0.885) reliability of affective disorder and neurodevelopment disorder. The convergent validity of affective disorder and neurodevelopment disorder was good, and their divergent validity was acceptable.Limitations: i) Clinical questionnaires were insensitive in classifying disorders and had limitations in derived diagnoses. ii) Samples only came from clinical environment, iii) covered limited disease species, and iv) were small.Conclusion: The K-SADS-PL-C DSM-5 can support reliable and valid diagnoses for children with affect, neurodevelopmental, and behavioral disorders in China. [ABSTRACT FROM AUTHOR]

Objective: Effects of analgesia effect of ambulatory labor analgesia and its influence on the mother and fetus. Methods: A retrospective analysis of 177 cases of pregnant women in our hospital from September 2012 to September 2013. According to the mode of delivery, the cases were divided into painless delivery group (n=83) and conventional delivery group (n=94), and compared the two groups in the labor pain score, birth process and newborn situation. Results: Cesarean section rate was significantly lower in painless childbirth group than that in conventional delivery group; the first stage in painless childbirth group is shorter than the conventional delivery group,the second stage of labor was significantly prolonged compared with that in the conventional delivery group (P<0.05), third stage has no significant statistical difference in two groups (P>0.05); painless childbirth pregnant woman the production process of VAS scores were significantly lower than that of conventional delivery group (P<0.05); and the incidence of neonatal Apgar score of two groups of fetal distress were no significant statistical difference (P>0.05); the two group of postpartum hemorrhage and the incidence of bleeding volume were not significantly difference (P>0.05); conventional delivery group no untoward reaction of postpartum, painless labor group 4 cases of orthostatic hypotension (5.6%), 3 cases of lower extremity muscle weakness (4.2%), were after symptomatic treatment in 3 h mitigation. Conclusions: Ambulatory labor analgesia can significantly reduce maternal pain of the production process, and will not lead to severe postpartum complications, fetal harm, is an effective, safe childbirth analgesia method, is worthy of clinical application. [ABSTRACT FROM AUTHOR]

Abstract: Objective: The ω-3 polyunsaturated fatty acids (PUFAs) play a key role as immune response modulators and suppressors of immunologic functions, such as lymphocyte proliferation, cytokine production, and cell surface molecular expression in T lymphocytes, monocytes, and natural killer cells. However, little is known about the effect of ω-3 PUFAs on dendritic cells (DCs). We studied the effect of ω-3 PUFAs on DCs and the related intracellular signal transduction pathway. Methods: Dendritic cells were generated from human peripheral blood monocytes in the presence of granulocyte-macrophage colony-stimulating factors and interleukin (IL)-4 and treated with eicosapentaenoic acid (EPA), docosahexanoic acid (DHA), and stearic acid for 24 h. Lipopolysaccharide (LPS) was used for maturation of the DCs. The expressions of CD40, CD80, CD86, and human leukocyte antigen-DR (HLA-DR) were analyzed by flow cytometry; production of IL-12 and tumor necrosis factor-α were detected by reverse transcriptase-polymerase chain reaction and enzyme-linked immunosorbent assay. The proliferative ability of allogeneic T cells stimulated by DCs was evaluated by tritiated thymidine (3H-TdR). Western blot analysis of p38 mitogen-activated protein kinase was conducted. Results: The ω-3 PUFAs reduced expression levels of costimulatory molecules CD80 and CD86 and major histocompatibility complex HLA-DR. IL-12 and tumor necrosis factor-α levels decreased significantly in the EPA and DHA groups. EPA and DHA also significantly reduced the proliferative ability of allogeneic T cells stimulated by DCs. The ω-3 PUFAs significantly inhibited LPS-induced p38 phosphorylation. Conclusion: The ω-3 PUFAs may inhibit LPS-induced DC maturation and upregulate cytokine production. Impaired p38 mitogen-activated protein kinase activity is a potential critical intracellular signaling transduction mechanism. [Copyright &y& Elsevier]

Most studies of mixed culture fermentation for hydrogen production focus on the metabolite distribution under different operational conditions. While, the corresponding microbial dynamics research is limited. The aim of this work was to reveal the microbial community dynamics of extreme-thermophilic mixed culture enriched in a chemostat for hydrogen production with the combining molecular tools of denaturing gradient gel electrophoresis (DGGE), Illumina Miseq high-throughput sequencing platform, and 16S rRNA clone library sequencing. The analyzed results showed that the genera of Caldanaerobius, Caldicellulosiruptor and/or Thermoanaerobacter dominated the bacteria. While archaea were not detected even by Illumina Miseq high-throughput sequencing. The effects of hydrogen partial pressure or pH increasing from 5.5 to 7.0 on the microbial communities were not notably, and genera of Caldicellulosiruptor and Thermoanaerobacter were the dominants. High hydrogen yield in this chemostat was attributed to the enrichment of Caldicellulosiruptor . While Caldanaerobius dominated the bacteria communities as pH decreased to 4.5. Caldanaerobius and Thermoanaerobacter dominated at high influent glucose concentrations. Thereby, it clearly indicated that the microbial dynamics shall be considered when analyzing the extreme-thermophilic mixed culture fermentation in the future. [ABSTRACT FROM AUTHOR]

The microbial community compositions of a chemostat enriched in a thermophilic (55 °C) mixed culture fermentation (MCF) for hydrogen production under different operational conditions were revealed in this work by integrating denaturing gradient gel electrophoresis (DGGE), Illumina Miseq high-throughput sequencing, and 16S rRNA clone library sequencing. The results showed that the community structure of the enriched cultures was relatively simple. Clones close to the genera of Thermoanaerobacter and/or Bacillus mainly dominated the bacteria. And homoacetogens and archaea were washed out and not detected even by Illumina Miseq high-throughput sequencing which supported the benefit for hydrogen production. On the other hand, the results revealed that the metabolic shift was clearly associated with the change of dominated bacterial groups. The effects of hydrogen partial pressure (P) and pH from 4.0 to 5.5 on the microbial compositions were not notable and Thermoanaerobacter was dominant, thus, the metabolites were also not changed. While Bacillus, Thermoanaerobacter and Propionispora hippei dominated the bacteria communities at neutral pH, or Bacillus and Thermoanaerobacter dominated at high influent glucose concentrations, consequently the main metabolites shifted to acetate, ethanol, propionate, or lactate. Thereby, the effect of microbial composition on the metabolite distribution and shift shall be considered when modeling thermophilic MCF in the future. [ABSTRACT FROM AUTHOR]

Immune dysfunction is a major cause of mortality in septic patients. Current evidence indicates an important role for dendritic cells (DCs) in the pathophysiology of immune dysfunction, and these cells are potential targets of immunomodulation therapies. In the present study, our aim was to enhance the resistance of endotoxemic mice to bacterial translocation and secondary infection and to improve the outcome of these infections using a combination therapy consisting of thymosin alpha1 and dexamethasone in a timely manner according to the changes of DCs' number. The effect of treatment with dexamethasone (DXM) and thymosin alpha1 (Tα1) on DCs was investigated by examining their number, MHCII and CD86 expression and their capacity to induce T cell activation. Endotoxemic mice were randomly divided into five treatment groups. The survival rates, the levels of TNF-α and IL-10, the occurrence of bacterial translocation, and the ability to clear secondary infections were determined. Additionally, the behavior of DCs over time was also evaluated. Tα1 induced significant increases in DC numbers in vivo, whereas DXM reduced cell numbers both in vitro and in vivo. However, neither drug induced significant changes in the capacity of DCs to induce T cell activation or their expression of MHCII or CD86. Among the five treatment groups, the mice treated with a combination of DXM and Tα1 had the highest survival rate; this increased survival was associated with a decrease in bacterial translocation to extra-intestinal organs and an enhanced ability to eradicate secondary infections by reversing the change in DC numbers during endotoxemia. Immunomodulatory therapy that combines Tα1 and DXM in a timely manner and was based on changes in DCs enhanced the resistance of endotoxemic mice to bacterial translocation and secondary infections, improving the outcome of the infection. [ABSTRACT FROM AUTHOR]

Background: Gut barrier failure caused by endotoxemia is a life-threatening problem. The present study aimed to determine whether any specific intestinal site is highly correlated with gut barrier failure, and whether recombinant human growth hormone (rhGH) can ameliorate gut barrier failure in a rat model of endotoxemia. Methods: Enterostomy tubes were surgically placed in adult male Sprague-Dawley rats three days before induction of endotoxemia by lipopolysaccharide (LPS) injection. Controls received no LPS. Rats were then randomly assigned to receive subcutaneous injections of rhGH (experimental, n = 30) or 0.9 % saline (control, n = 15) at 24, 48, or 72 h after LPS injection. Escherichia coli labeled with green fluorescent protein (GFP) were injected into the intestinal segment of all rats through the enterostomy tubes. The number of GFP-labeled E. coli detected in mesenteric lymph nodes was examined after 96 h. Apoptosis and proliferation rates of intestinal epithelial cells, and intestinal permeability were measured. Results: Endotoxemia led to high mortality, compared with the control group, and rhGH treatment did not improve survival. Intestinal permeability, reflected by translocation rates of GFP-labeled E. coli, and apoptosis rates in the LPS-induced endotoxemia group were higher than those in the non-endotoxemia control group, and the endotoxemia ileum group had the highest rates of both bacterial translocation and apoptosis. The LPS+GH group had less bacterial translocation and apoptosis than the LPS-induced endotoxemia group. In contrast, the proliferation rates were lower in the LPS group compared to the LPS+GH group. Conclusions: Endotoxemia can induce gut barrier failure in rats, and the ileum is the site of greatest risk. The GH can reduce the incidence of endotoxemia-induced gut barrier failure, but not the associated mortality. [ABSTRACT FROM AUTHOR]

Abstract: Berberine hydrochloride (BBR), a plant alkaloid, has been used to treat intestinal inflammation or infection for years. Cyclooxygenase-2 (COX-2) is pro-inflammatory mediator and involved in the induction of gut inflammation. The expression of COX-2 in small bowel mucosa was determined and the mechanism by which BBR modulated COX-2 expression was explored in a rat model of endotoxemia induced by lipopolysaccharide (LPS). The results showed that without LPS stimulation COX-2 was constitutively expressed at low levels in control rats. LPS challenge rapidly induced COX-2 gene transcription resulting in high levels of inducible COX-2 expression in endotoxemic rats. BBR pre- and post-treatment had no marked effect on constitutive COX-2 expression but inhibited inducible COX-2 overexpression. LPS challenge increased the expression and phosphorylation of peroxisome proliferator-activated receptor gamma (PPARγ), p38 and activating transcription factor 2 and 3 (ATF2, ATF3), but the effects of LPS were inhibited by BBR treatment. GW9662 did not influence constitutive COX-2 expression but enhanced inducible COX-2 overproduction. Besides, GW9662 abolished the inhibitory effect of BBR on inducible COX-2, p38, ATF2, 3 expression and phosphorylation. Collectively, these results indicated that BBR gavage could attenuate the overexpression of inducible COX-2, not constitutive COX-2, in ileal mucosa during acute endotoxemia in part via activation of PPARγ pathway, which negatively interfered with p38/ATFs cascade. [Copyright &y& Elsevier]

Abstract: The effect of berberine hydrochloride (BBR) on inducible cyclooxygenase-2 (COX-2) in small intestinal mucosa and related mechanisms was investigated in a rat model of acute endotoxemia. The results showed that lipopolysaccharide (LPS) increased COX-2 expression, whereas SB202190 and BBR curtailed it. LPS increased phosphorylation of mucosal p38 MAPK and ATF2 as well as production of ATF2, whereas BBR attenuated these effects. LPS upregulated mucosal peroxisome proliferator-activated receptor gamma (PPARγ), but BBR reduced this receptor. GW9662 aggravated LPS-induced and reversed BBR-attenuated COX-2 expression. The findings showed that BBR ameliorated COX-2 overexpression partially via modulation of p38 and PPARγ pathways during acute endotoxemia. [Copyright &y& Elsevier]

Vernoramyosides A-F (1 – 6), six new Δ7,9(11) stigmastane-type steroid saponins, along with four known analogues (7 – 10) were isolated from the leaves of Vernonia amygdalina Delile (Compositae). Their structures were determined by the combination of NMR, ECD and HR-ESI-MS data. These compounds all possessed highly oxidized side chain and a γ-lactam or α , β -unsaturated five-membered lactone ring. All isolates were screened for their activities in reversing resistance in MCF/DOX cells. [Display omitted] [ABSTRACT FROM AUTHOR]