Discovery of natural AMPK activator from the fruits of Xanthium sibiricum Patr.: Xanthiumine A, protoberberine alkaloid with unique C28 skeleton.

Two protoberberine alkaloids with a unique C28 skeleton, named xanthiumines A (1) and B (2), respectively, were isolated from the fruits of Xanthium sibiricum Patr. Their structures including absolute configurations were unequivocally established by the comprehensive NMR and MS spectroscopic data analysis together with gauge-independent atomic orbital (GIAO) NMR calculations, and electronic circular dichroism (ECD) calculations. Compounds 1 and 2 are the first examples of natural protoberberine alkaloid with a phenolic acid group at C-13a. Their plausible biosynthetic pathway was proposed on the basis of the coexisting alkaloid monomer as the precursor. Furthermore, the effects and related molecular mechanism of compound 1 on hepatic lipid accumulation were also investigated in oleic acid (OA)-treated HepG2 cells.

Synthetic Xylosides: Probing the Glycosaminoglycan Biosynthetic Machinery for Biomedical Applications.

Glycosaminoglycans (GAGs) are polysaccharides ubiquitously found on cell surfaces and in the extracellular matrix (ECM). They regulate numerous cellular signaling events involved in many developmental and pathophysiological processes. GAGs are composed of complex sequences of repeating disaccharide units, each of which can carry many different modifications. The tremendous structural variations account for their ability to bind many proteins and thus, for their numerous functions. Although the sequence of GAG biosynthetic events and the enzymes involved mostly were deduced a decade ago, the emergence of tissue or cell specific GAGs from a nontemplate driven process remains an enigma. Current knowledge favors the hypothesis that macromolecular assemblies of GAG biosynthetic enzymes termed "GAGOSOMEs" coordinate polymerization and fine structural modifications in the Golgi apparatus. Distinct GAG structures arise from the differential channeling of substrates through the Golgi apparatus to various GAGOSOMEs. As GAGs perform multiple regulatory roles, it is of great interest to develop molecular strategies to selectively interfere with GAG biosynthesis for therapeutic applications. In this Account, we assess our present knowledge on GAG biosynthesis, the manipulation of GAG biosynthesis using synthetic xylosides, and the unrealized potential of these xylosides in various biomedical applications. Synthetic xylosides are small molecules consisting of a xylose attached to an aglycone group, and they compete with endogenous proteins for precursors and biosynthetic enzymes to assemble GAGs. This competition reduces endogenous proteoglycan-bound GAGs while increasing xyloside-bound free GAGs, mostly chondroitin sulfate (CS) and less heparan sulfate (HS), resulting in a variety of biological consequences. To date, hundreds of xylosides have been published and the importance of the aglycone group in determining the structure of the primed GAG chains is well established. However, the structure-activity relationship has long been cryptic. Nonetheless, xylosides have been designed to increase HS priming, modified to inhibit endogenous GAG production without priming, and engineered to be more biologically relevant. Synthetic xylosides hold great promise in many biomedical applications and as therapeutics. They are small, orally bioavailable, easily excreted, and utilize the host cell biosynthetic machinery to assemble GAGs that are likely nonimmunogenic. Various xylosides have been shown, in different biological systems, to have anticoagulant effects, selectively kill tumor cells, abrogate angiogenic and metastatic pathways, promote angiogenesis and neuronal growth, and affect embryonic development. However, most of these studies utilized the commercially available one or two ß-D-xylosides and focused on the impact of endogenous proteoglycan-bound GAG inhibition on biological activity. Nevertheless, the manipulation of cell behavior as a result of stabilizing growth factor signaling with xyloside-primed GAGs is also reckonable but underexplored. Recent advances in the use of molecular modeling and docking simulations to understand the structure-activity relationships of xylosides have opened up the possibility of a more rational aglycone design to achieve a desirable biological outcome through selective priming and inhibitory activities. We envision these advances will encourage more researchers to explore these fascinating xylosides, harness the GAG biosynthetic machinery for a wider range of biomedical applications, and accelerate the successful transition of xyloside-based therapeutics from bench to bedside.

Bullying in professional sports: psychological needs of athletes.

BACKGROUND: This paper aimed to establish whether bullying in sports affects the satisfaction of such psychological needs as autonomy, competence, and relatedness in professional sports. METHODS: The instruments in this work were the Bullying Participant Behaviors Questionnaire (BPBQ), the Motivational Mediators Scale in Sport (EMMD), and the Psychological Needs Thwarting Scale (PNTS). The participants were 708 professional athletes. RESULTS: Comparison of EMMD and PNTS means unveiled that professional athletes with no bullying experience are more psychologically satisfied and less thwarted in all three dimensions (competence, autonomy, and relatedness). Among the group exposed to bullying, victims (18.92) and bullies (23.18) had the lowest needs in terms of competence, while bullies (26.14) and victims (20.10) experienced the lowest autonomy. The relatedness factor was most pronounced in victims' defenders (34.06) and least in victims (16.39). The lowest competence thwarting was found for outsiders and defenders, and the highest - among victims of bullying (18.12). But both bullies and their helpers had significantly higher scores than the other two roles. The need for autonomy, in turn, was least thwarted in outsiders and defenders, and most - in victims, as in the case of the relatedness subscale. CONCLUSIONS: The practical and scientific value of this work stem from the fact that it proves the negative impact of bullying on the satisfaction of basic psychological needs. The obtained findings can facilitate the development and implementation of updated educational programs and practices, leadership systems, as well as be conducive to the work of sports psychologists.

Methods for Assessing the Effects of Xylosides on Angiogenesis.

Xylosides are small synthetic molecules consisting of a xylose molecule attached to an aglycone group and serve as primers in the assembly of core protein free glycosaminoglycans using cellular machinery. Synthetic xylosides hold great promise in many biomedical applications and as therapeutics. Recent advances in the study of xylosides have opened up the possibility of developing xylosides as therapeutics to achieve a desirable biological outcome through their selective priming and inhibitory activities toward glycosaminoglycan biosynthesis. The approach described, herein, will serve as a general strategy to comprehensively screen xylosides and evaluate their ability to promote or inhibit angiogenesis, a critical biological process that is dysregulated in over 70 human diseases.

Manipulation of Glycosaminoglycans Using Synthetic Xylosides to Study Their Roles in Lung Branching Morphogenesis in Ex Vivo Lung Bud Culture System.

The primary left and right bronchial buds grow and sprout secondary bronchi, which in turn develop tertiary bronchi, and so on. Branching continues for a total of 6-8 generations in the mouse and for about 23 generations in humans, forming the estimated 50 million branches of the human lung. Thus, patterns of branching are incalculably complex. However, these branches are rarely random, implying that they are under genetic control. Genomic information alone cannot specify the patterning information in terms of where the branching occurs and the direction it grows as well as their size and shape. There is a complex choreography among glycosaminoglycans and growth factors/morphogens that provide a highly complex instructive cues that control lung branching and development of the functional lung. Herein, we describe the use of xylosides in the manipulation of glycosaminoglycan (GAG) biosynthesis and study the effect of xyloside-primed GAGs in the regulation of lung branching events.

Applications of Xylosides in the Manipulation of Stem Cell Niche to Regulate Human Neural Stem Cell Differentiation and Neurite Outgrowth.

The extracellular matrix (ECM) plays a pivotal role in the regulation of neural stem cell differentiation, axon guidance and growth, and neural plasticity. Glycosaminoglycans, such as heparan sulfate and chondroitin sulfate, are significant components of brain ECM that dictates neurogenesis and neural repair. Herein, we describe a simple method to assess the effect of xylsoides, which serve as primers and inhibitors of GAG biosynthesis, on human neural stem cell differentiation and neurite outgrowth in in vitro culture conditions.

Methods to Analyze the Effect of Diet-Derived Metabolites on Endothelial Inflammation and Cell Surface Glycosaminoglycans.

The glycocalyx is a biologically active barrier that covers the luminal side of the vascular endothelium and it is comprised of proteoglycans [core proteins with glycosaminoglycans (GAG) side chains], glycoproteins, and plasma proteins. Evidence shows that the disruption in the structure and function of the endothelial glycocalyx exacerbates vascular inflammation and atherosclerosis. The GAG components of the glycocalyx undergo remodeling in the setting of diabetes and these alterations in endothelial GAGs negatively impact the vascular function. Hence, the preservation and restoration of GAGs in altered vasculature may be a novel strategy to ameliorate vascular complications in diabetes and metabolic syndrome. Human studies support the beneficial vascular effects of flavonoids which are widely found in fruits and vegetables. Flavonoids are extensively metabolized by the intestinal microbiota and digestive enzymes in humans, suggesting that their biological activities may be mediated by their circulating metabolites. Studies indicate that counteracting the damage to GAGs using dietary compounds improve vascular complications. In this article, we describe the methods to analyze the effect of diet-derived metabolites such as metabolites of flavonoids on endothelial inflammation and cell surface glycosaminoglycans.

Cu-Co Co-Doped Microporous Coating on Titanium with Osteogenic and Antibacterial Properties.

Titanium (Ti) and its alloys are widely used in bone surgery by virtue of their excellent mechanical properties and good biocompatibility; however, complications such as loosening and sinking have been reported post-implantation. Herein we deposited a copper-cobalt (Cu-Co) co-doped titanium dioxide (TUO) coating on the surface of Ti implants by microarc oxidation. The osteogenic and antimicrobial properties of the coating were evaluated by in vitro experiments, and we also assessed ß-catenin expression levels on different sample surfaces. Our results revealed that the coating promoted the adhesion, proliferation, and differentiation of MG63 osteoblasts, and TUO coating promoted ß-catenin expression; moreover, the proliferation of Staphylococcus aureus was inhibited. To summarize, we report that Cu-Co co-doping can enhance the osteogenic and antibacterial activities of orthopedic Ti implants, leading to potentially improved clinical performance.

Residue analysis of acephate and its metabolite methamidophos in open field and greenhouse pakchoi (Brassica campestris L.) by gas chromatography-tandem mass spectrometry.

To analyze the dynamic degradation and final residues of acephate and its metabolite methamidophos, field-experiments with pakchoi (Brassica campestris L.) in open field and greenhouse were carried out in Beijing, China in 2004 and 2005. The degradation dynamics and final residues were determined by gas chromatography (GC) equipped with a pulsed flame photometric detector and GC coupled to mass spectrometry (MS)/MS after acephate was applied on open field and green house pakchoi (B. campestris L.). The dynamic degradation results showed that the half-lives of acephate and methamidophos in open field pakchoi were 1.36 days with dynamic degradation equation C( t ) = 133.01e( - 0.5107t ), and 2.86 days with C( t ) = 6.5753e( - 0.2422t ), respectively. While the half-lives of acephate and methamidophos in the greenhouse were 1.07 days with C( t ) = 59.134e( - 0.4353t ) and 0.79 days with C( t ) = 0.2703e( - 0.2595t ), respectively. The final residue analysis demonstrated that >50% of total methamidophos were resulted from the degradation of acephate 7 and 18 days after it was applied on the greenhouse pakchoi, respectively. While in the open-field pakchoi, >90% of total methamidophos was found to be the metabolite of acephate.

Genetic diversity and genetic differentiation of invasive weed Xanthium italicum in China.

Xanthium italicum is an aggressive weed found worldwide. Despite several ecological, morphological, and physiological research on its invasion mechanism, the mechanism of its successful invasion has not been revealed from the viewpoint of population genetics. Thus, we aimed to evaluate the genetic variation within and among populations of the alien invasive weed X. italicum in China, and to provide a theoretical basis for its invasion mechanism. For that, we employed inter-simple sequence repeat (ISSR) markers to explore the genetic diversity and genetic differentiation of 185 individuals sampled from 10 populations. Eight selected primers yielded a total of 76 bright and discernible bands. X. italicum showed an intermediate genetic diversity at the population level (percentage of polymorphic loci (PPL) = 60.26%, Nei's genetic diversity (H) = 0.2098, Shannon's information index (I) = 0.3129). However, the genetic diversity at the species level was significantly high (PPL = 100%; H = 0.3673; I = 0.5425). The coefficient of gene differentiation (GST, 41.4%) and analysis of molecular variance showed that genetic differentiation mainly occurred within populations. The estimated gene flow (Nm, 0.7085) and Mantel test indicated that genetic differentiation in the populations may primarily come from genetic drift and anthropogenic activities. Our results revealed the high genetic diversity of X. italicum, which may help explain its invasion success in China. This knowledge may contribute to the efforts for decreasing and eventually stopping X. italicum invasion in China.

Xanthium italicum est une plante envahissante trouvée dans le monde entier. En dépit de quelques recherches écologiques, morphologiques et physiologiques à propos de son mécanisme d'invasion, le mécanisme de son invasion réussie n'a pas encore été révélé du point de vue de la génétique démographique. Donc, nous avons visé à évaluer la variation génétique au sein de et parmi les populations de plante exotique envahissante X. italicum en Chine, et à offrir une base théorique à son mécanisme d'invasion. À cet effet, nous avons employé des marqueurs des répétitions de séquences inter-simples (ISSR) afin d'explorer la diversité génétique et la différenciation génétique de 185 individus échantillonnés à partir de 10 populations. Huit amorces sélectionnées ont donné un total de 76 bandes brillantes et perceptibles. X. italicum a montré une diversité génétique intermédiaire au niveau de la population (pourcentage de loci polymorphe (PPL) = 60.26%, la diversité génétique de Nei (H) = 0.2098, l'indice d'information de Shannon (I) = 0.3129). Toutefois, la diversité génétique était considérablement élevée au niveau des espèces (PPL = 100% ; H = 0.3673 ; I = 0.5425). Tant le coefficient de différenciation génétique (GST, 41.4%) que l'analyse de la variance moléculaire ont reflété que la différenciation génétique se produisait principalement au sein des populations. En fonction du flux génétique estimé (Nm, 0.7085) et du test Mantel, la différenciation génétique au sein des populations pourrait provenir principalement de la dérive génétique et des activités anthropiques. Nos résultats ont révélé la haute diversité génétique de X. italicum, cela peut aider à expliquer son succès d'invasion en Chine. Ces connaissances pourraient contribuer à la réduction et à l'arrêt éventuel de l'invasion de X. italicum en Chine.

Visualizing antithrombin-binding 3-O-sulfated heparan sulfate motifs on cell surfaces.

To map the cellular topography of the rare 3-O-sulfated structural motif of heparan sulfate (HS), we constructed quantum dot-based probes for antithrombin and FGF2, which reveal widely different distribution of the targeted HS motifs. The technology helps show that old and young aortic endothelia display widely different levels of the antithrombin-binding 3-O-sulfated HS motif.

Prevalence of abnormity of blood lipid and associated factors in health examination population in Beijing.

OBJECTIVE: To investigate the prevalence of abnormity of blood lipid and associated factors in healthy population in Beijing. METHODS: Totally, 38462 individuals who received health examination were enrolled in our study. We divided them into eight groups according to their ages. The levels of serum total cholesterol, triglyceride, high density lipoprotein cholesterol, and low density lipoprotein cholesterol were tested, and the relationship of blood lipid abnormity with body mass index (BMI) and fasting blood glucose was analyzed. RESULTS: The incidences of hypercholesterolemia, hyperglyceridemia, low high-density lipoprotein cholesterolemia, and hyper low-density lipoprotein cholesterolemia presented increasing trend in this population. The incidence rate of abnormity of blood lipid in health examination population increased with BMI increase. The incidence of abnormity of blood lipid in overweight and obesity population was significantly higher than that in low weight and normal weight populations (P<0.05). Meanwhile, the trend of abnormal blood lipid incidence coincided with that of abnormal fasting blood glucose. CONCLUSIONS: The prevalence of overweight, obesity, and abnormity of blood lipid in Beijing presents increasing trend. The incidence of abnormity of blood lipid increases with BMI increase, in coincidence with that of fasting blood glucose.

Prevalence of metabolic syndrome and its associations with other metabolic disorders and cardiovascular changes in health examination population in Beijing.

OBJECTIVE: To investigate the prevalence of metabolic syndrome (MS) and its associations with other metabolic disorders and cardiovascular changes in health examination population in Beijing. METHODS: Totally, 10,916 individuals who received health examination in Health Examination Center of Peking Union Medical College Hospital were enrolled. The height, weight, blood pressure, serum levels of triglyceride, high-density lipoprotein cholesterol (HDL-C), and fasting blood glucose were recorded. MS was diagnosed based on the working criteria of Chinese Diabetes Society 2004 (CDS2004). Meanwhile, other metabolic disorders, including fatty liver and hyperuricemia, were recorded. The cardiovascular changes were reflected by the reports of electrocardiogram (ECG) ST-T changes and atherosclerosis of retinal arteries. RESULTS: The overall prevalence rate of MS was 6.1% (666/10,916) in the population. The prevalence rate of MS in male was much higher than that in female (9.0% vs. 2.7%, P=0.000). For individuals with MS, the prevalence rates of fatty liver and hyperuricemia were significantly higher than those without MS, respectively (70.4% vs. 35.4%, P=0.000; 29.9% vs. 17.7%, P=0.000). As for cardiovascular changes, the prevalence rates of ECG ST-T changes and atherosclerosis of retinal arteries were significantly higher in individuals with MS than those without MS, respectively (13.8% vs. 11.7%, P=0.012; 12.0% vs. 6.8%, P=0.000). CONCLUSIONS: The prevalence of MS in Beijing population is high. The individuals with MS have a higher risk for other metabolic disorders and cardiovascular changes.

Analysis of interleukin-17 and interleukin-23 for estimating disease activity and predicting the response to treatment in active lupus nephritis patients.

Renal biopsy is a "gold standard" for establishing the diagnosis and assessing prognosis and monitoring therapy in lupus nephritis (LN) patients, but it is an invasive and inconvenient procedure. Evidences showed that interleukin-17(IL-17) and interleukin-23(IL-23) may be as alternative biomarkers for diagnosing LN, monitoring LN activity and predicting the response to treatment of LN. To analyze the roles of IL-17 and IL-23 in evaluation activity of LN and predicting active LN response to immunosuppressive treatment, by comparison between IL-17, IL-23 and clinical data of LN. Eighty patients with LN and 20 healthy volunteers were enrolled in this study. Plasma levels of IL-17 and IL-23 were detected by ELISA and clinical data were collected in patients with LN. Thirty-seven patients with active LN accepted immunosuppressive therapy and followed up to 6 months. The roles of IL-17 and IL-23 in evaluation the activity of LN and the predictability for active LN response to immunosuppressive treatment were analyzed. The ages or gender rations between LN patients and healthy controls were not significant difference at baseline. Baseline levels of IL-17 and IL-23 were higher in patients with active LN compare to them in patients with inactive LN or controls (P<0.001) and IL-23 in patients with inactive LN was higher than its in controls (P=0.004). IL-17 and IL-23 decreased significantly in active LN patients after 6 months therapy (P<0.001). The baseline level of IL-23 was significantly different in subgroups response to the immunosuppressive treatment in patients with active LN (P=0.0014). Baseline level of IL-23 in complete response group was lower than its in partial response group (P=0.0015) or nonresponse group (P=0.013). IL-17 was negative correlation with C3 (r=-0.44, P<0.001). IL-17 and IL-23 correlated with systemic lupus erythematosus (SLE) disease activity index (P<0.001). The correlation between IL-17 and LN pathological acute index (AI) was higher than the correlation between IL-23 and AI. (r=0.52, P<0.001 vs. r=0.41, P<0.001). Receiver Operation Characteristics (ROC) showed that IL-17 and IL-23 could be used to evaluate SLE disease activity index. IL-17 could be used as biomarker to evaluate pathological AI. IL-23 could be used as a predictor for predicting response to immunosuppressive treatment in patients with active LN. IL-17 and IL-23 may involve and contribute to LN. IL-17 could be used as a biomarker for LN clinical and pathological AI. IL-23 could be used as a predictor for predicting response to immunosuppressive treatment in patients with active LN.

Effect of oxymatrine on Cryptosporidium parvum infection in mice based on the HMGB1-TLR2/TLR4-NF-κB pathway / 中国血吸虫病防治杂志

Objective To investigate the involvement of the high mobility group box protein B1 (HMGB1)-Toll-like receptor 2 (TLR2)/TLR4-nuclear factor κB (NF-κB) pathway in the intestinal mucosal injury induced by Cryptosporidium parvum infection, and to examine the effect of oxymatrine (OMT) on C. parvum infection in mice. Methods Forty SPF 4-week-old BALB/c mice were randomly divided into four groups, including the control group, infection group, glycyrrhizin (GA) group and OMT group. Each mouse was orally administered with 1 × 105 C. parvum oocysts one week in the infection, GA and OMT groups following dexamethasone-induced immunosuppression to model C. parvum intestinal infections in mice. Upon successful modeling, mice in the GA group were intraperitoneally injected with GA at a daily dose of 25.9 mL/kg for successive two weeks, and animals in the OMT group were orally administered OMT at a daily dose of 50 mg/kg for successive two weeks, while mice in the control group were given normal food and water. All mice were sacrificed two weeks post-treatment, and proximal jejunal tissues were sampled. The pathological changes of mouse intestinal mucosal specimens were observed using hematoxylin-eosin (HE) staining, and the mouse intestinal villous height, intestinal crypt depth and the ratio of intestinal villous height to intestinal crypt depth were measured. The occludin and zonula occludens protein 1 (ZO1) expression was determined in mouse intestinal epithelial cells using immunohistochemistry, and the relative expression of HMGB1, TLR2, TLR4, myeloid differentiation primary response gene 88 (MyD88) and NF-κB p65 mRNA was quantified in mouse jejunal tissues using quantitative real-time PCR (qPCR) assay. Results HE staining showed that the mouse intestinal villi were obviously atrophic, shortened, and detached, and the submucosal layer of the mouse intestine was edematous in the infection group as compared with the control group, while the mouse intestinal villi tended to be structurally intact and neatly arranged in the GA and OMT groups. There were significant differences among the four groups in terms of the mouse intestinal villous height (F = 6.207, P = 0.000 5), intestinal crypt depth (F = 6.903, P = 0.000 3) and the ratio of intestinal villous height to intestinal crypt depth (F = 37.190, P < 0.000 1). The mouse intestinal villous height was lower in the infection group than in the control group [(321.9 ± 41.1) μm vs. (399.5 ± 30.9) μm; t = 4.178, P < 0.01] and the GA group [(321.9 ± 41.1) μm vs. (383.7 ± 42.7) μm; t = 3.130, P < 0.01], and the mouse intestinal crypt depth was greater in the infection group [(185.0 ± 35.9) μm] than in the control group [(128.4 ± 23.6) μm] (t = 3.877, P < 0.01) and GA group [(143.3 ± 24.7) μm] (t = 2.710, P < 0.05). The mouse intestinal villous height was greater in the OMT group [(375.3 ± 22.9) μm] than in the infection group (t = 3.888, P < 0.01), and there was no significant difference in mouse intestinal villous height between the OMT group and the control group (t = 1.989, P > 0.05). The mouse intestinal crypt depth was significantly lower in the OMT group [(121.5 ± 27.3) μm] than in the infection group (t = 4.133, P < 0.01), and there was no significant difference in mouse intestinal crypt depth between the OMT group and the control group (t = 0.575, P > 0.05). The ratio of the mouse intestinal villous height to intestinal crypt depth was significantly lower in the infection group (1.8 ± 0.2) than in the control group (3.1 ± 0.3) (t = 10.540, P < 0.01) and the GA group (2.7 ± 0.3) (t = 7.370, P < 0.01), and the ratio of the mouse intestinal villous height to intestinal crypt depth was significantly higher in the OMT group (3.1 ± 0.2) than in the infection group (t = 15.020, P < 0.01); however, there was no significant difference in the ratio of the mouse intestinal villous height to intestinal crypt depth between the OMT group and the control group (t = 0.404, P > 0.05). Immunohistochemical staining showed significant differences among the four groups in terms of occludin (F = 28.031, P < 0.000 1) and ZO1 expression (F = 14.122, P < 0.000 1) in mouse intestinal epithelial cells. The proportion of positive occluding expression was significantly lower in mouse intestinal epithelial cells in the infection group than in the control group [(14.3 ± 4.5)% vs. (28.3 ± 0.5)%; t = 3.810, P < 0.01], and the proportions of positive occluding expression were significantly higher in mouse intestinal epithelial cells in the GA group [(30.3 ± 1.3)%] and OMT group [(25.8 ± 1.5)%] than in the infection group (t = 7.620 and 5.391, both P values < 0.01); however, there was no significant differences in the proportion of positive occluding expression in mouse intestinal epithelial cells between the GA or OMT groups and the control group (t = 1.791 and 2.033, both P values > 0.05). The proportion of positive ZO1 expression was significantly lower in mouse intestinal epithelial cells in the infection group than in the control group [(14.4 ± 1.8)% vs. (24.2 ± 2.8)%; t = 4.485, P < 0.01], and the proportions of positive ZO1 expression were significantly higher in mouse intestinal epithelial cells in the GA group [(24.1 ± 2.3)%] (t = 5.159, P < 0.01) and OMT group than in the infection group [(22.5 ± 1.9)%] (t = 4.441, P < 0.05); however, there were no significant differences in the proportion of positive ZO1 expression in mouse intestinal epithelial cells between the GA or OMT groups and the control group (t = 0.037 and 0.742, both P values > 0.05). qPCR assay showed significant differences among the four groups in terms of HMGB1 (F = 21.980, P < 0.000 1), TLR2 (F = 20.630, P < 0.000 1), TLR4 (F = 17.000, P = 0.000 6), MyD88 (F = 8.907, P = 0.000 5) and NF-κB p65 mRNA expression in mouse jejunal tissues (F = 8.889, P = 0.000 7). The relative expression of HMGB1 [(5.97 ± 1.07) vs. (1.05 ± 0.07); t = 6.482, P < 0.05] 、TLR2 [(5.92 ± 1.29) vs. (1.10 ± 0.14); t = 5.272, P < 0.05] 、TLR4 [(5.96 ± 1.50) vs. (1.02 ± 0.03); t = 4.644, P < 0.05] 、MyD88 [(3.00 ± 1.26) vs. (1.02 ± 0.05); t = 2.734, P < 0.05] and NF-κB p65 mRNA [(2.33 ± 0.72) vs. (1.04 ± 0.06); t = 2.665, P < 0.05] was all significantly higher in mouse jejunal tissues in the infection group than in the control group. A significant reduction was detected in the relative expression of HMGB1 (0.63 ± 0.01), TLR2 (0.42 ± 0.10), TLR4 (0.35 ± 0.07), MyD88 (0.70 ± 0.11) and NF-κB p65 mRNA (0.75 ± 0.01) in mouse jejunal tissues in the GA group relative to the control group (t = 8.629, 5.830, 11.500, 4.729 and 6.898, all P values < 0.05), and the relative expression of HMGB1, TLR2, TLR4, MyD88 and NF-κB p65 mRNA significantly reduced in mouse jejunal tissues in the GA group as compared to the infection group (t = 7.052, 6.035, 4.084, 3.165 and 3.274, all P values < 0.05). In addition, the relative expression of HMGB1 (1.14 ± 0.60), TLR2 (1.00 ± 0.24), TLR4 (1.14 ± 0.07), MyD88 (0.96 ± 0.25) and NF-κ B p65 mRNA (1.12 ± 0.17) was significantly lower in mouse jejunal tissues in the OMT group than in the infection group (t = 7.059, 5.320, 3.510, 3.466 and 3.273, all P values < 0.05); however, there were no significant differences between the OMT and control groups in terms of relative expression of HMGB1, TLR2, TLR4, MyD88 or NF-κB p65 mRNA in mouse jejunal tissues (t = 0.239, 0.518, 1.887, 0.427 and 0.641, all P values > 0.05). Conclusions C. parvum infection causes intestinal inflammatory responses and destruction of intestinal mucosal barrier through up-regulating of the HMGB1-TLR2/TLR4-NF-κB pathway. OMT may suppress the intestinal inflammation and repair the intestinal mucosal barrier through inhibiting the activity of the HMGB1-TLR2/TLR4-NF-κB pathway.

Mechanism of Qingfei Huatan Zhuyu Decoction in Treatment of COPD Based on "Lung and Intestinal Co-treatment" / 中国实验方剂学杂志

ObjectiveTo observe the effect of Qingfei Huatan Zhuyu decoction on the lung and intestinal function of rats with chronic obstructive pulmonary diseases （COPD） and explore the deep-seated mechanism of its embodiment of lung and intestinal co-treatment. MethodA total of 60 Wistar rats were randomly divided into six groups， with 10 rats in each group， and the groups were control group， model group， acute syrup group （10 g·kg-1·d-1）， and low， medium， and high-dose groups （10， 15， 20 g·kg-1·d-1） of Qingfei Huatan Zhuyu decoction. The COPD rat model was established by lipopolysaccharide tracheal drip combined with the smoke inhalation method， and the acute syrup group and the Qingfei Huatan Zhuyu decoction group were administered by gavage with corresponding dose concentrations respectively， while the rest groups were controlled by saline gavage， and the lung function and blood gas indexes of rats were monitored after the last administration. The histopathological changes in the lung and intestine were observed microscopically. The expression of serum interleukin-6 （IL-6）， tumor necrosis factor-α （TNF-α）， and secretory immunoglobulin A （IgA） in colon tissue were measured by enzyme-linked immunosorbent assay （ELISA）. The biochemical indexes such as serum diamine oxidase （DAO）， D-lactic acid， and malondialdehyde （MDA） were measured. Immunohistochemistry was used to detect the expression of tight junction protein （Occludin） in rat colon tissue. The expression of F4/80 positive alveolar macrophages in rat lung tissue， and the expression of α-actin （α-SMA） and colonic atresia small band protein-1 （ZO-1） were determined by immunofluorescence. The protein expression of p-NF-κB p65， NF-κB p65， p-p38 MAPK， and p-p38 MAPK and the expression of Occludin and ZO-1 in colon tissue were detected in rat lung tissue by Western blot. ResultCompared with the normal group， the model group had pulmonary dysfunction， reduced forced vital capacity （FVC）， arterial partial oxygen pressure （PaO2）， arterial oxygen saturation （SaO2）， and dynamic lung compliance （Cdyn） （P<0.01）， and the pathological changes in the lung and intestine were obvious. The expressions of IL-6， TNF-α， DAO， D-lactic acid， and MDA in serum were increased （P<0.05，P<0.01）， and the protein expression ratio of p-NF-κB p65/NF-κB p65 and p-p38 MAPK/p38 MAPK in lung tissue was increased. The expression of F4/80 positive macrophages in lung tissue was enhanced. The expression of IgA， Occludin， and ZO-1 in colon tissue decreased （P<0.05，P<0.01）. Compared with the model group， the pulmonary function of the rats in the acute syrup group and groups of Qingfei Huatan Zhuyu decoction was significantly improved， and the FVC， PaO2， SaO2， and Cdyn were increased （P<0.05， P<0.01）. The pathological changes in the lung and intestine were significant. The expressions of IL-6， TNF-α， DAO， D-lactic acid， and MDA in serum were decreased （P<0.05，P<0.01）， and the expressions of F4/80 positive macrophages in lung tissue were decreased （P<0.01）. The protein expression ratio of p-NF-κB p65/NF-κB p65 and p-p38 MAPK/p38 MAPK in lung tissue decreased （P<0.01）， and the expression of IgA， Occludin， and ZO-1 in colon tissue increased （P<0.01）. ConclusionQingfei Huatan Zhuyu decoction can effectively reduce the symptoms of COPD rats， and its mechanism of action is related to inhibiting the inflammatory response of lung tissue and improving the barrier function of the intestinal mucosa.

Effects of Zhoufei Pingchuan Capsules on Th17/Treg Balance and Related Inflammatory Factors in Peripheral Blood of Patients with Stable Chronic Obstructive Pulmonary Disease with Lung-kidney Qi Deficiency Syndrome / 中国中医药信息杂志

Objective To investigate the effects of Zhoufei Pingchuan Capsules on the balance of peripheral blood helper T lymphocyte 17 cell/regulatory T lymphocyte cell(Th17/Treg)and related inflammatory factors in peripheral blood of patients with stable chronic obstructive pulmonary disease(COPD)with lung-kidney qi deficiency syndrome.Methods Totally 40 COPD patients were randomly divided into the study group and the control group,with 20 cases in each group.Another 20 cases were in the healthy group.The control group was given tiotropium bromide powder inhalation,18 μg/time,1 time/d,inhalation;on the basis of the control group,the study group was given Zhoufei Pingchuan Capsules,3 pills/time,3 times/d,orally.All patients were treated for 8 weeks.The healthy group was not given any intervention.Forced expiratory volume in one second(FEV1),FEV1/forced vital capacity(FEV1/FVC),maximum mid-expiratory flow(MMEF),carbon monoxide diffusing capacity/alveolar ventilation(DLCO/VA),arterial partial pressure of oxygen(PaO2),arterial partial pressure of carbon dioxide(PaCO2),COPD assessment test(CAT)score,Th17/Treg ratio,cytokines interleukin(IL)-17,IL-22,IL-10,and transforming growth factor-β1(TGF-β1)were compared before and after treatment.Results Compared with before treatment,the lung function indexes(FEV1,FEV1/FVC,MMEF,DLCO/VA),blood gas indexes(PaO2,PaCO2)and CAT score in the study group after treatment were significantly improved(P<0.05).After treatment,the mean values of MMEF,DLCO/VA,PaCO2 and CAT score in the study group were better than those in the control group(P<0.05).Compared with before treatment,the levels of Th17,IL-17 and IL-22 in the study group were significantly lower,and the levels of Treg,IL-10 and TGF-β1 were significantly higher(P<0.05).After treatment,there were significant differences in Th17,Treg,IL-17,IL-22,IL-10 and TGF-β1 among the three groups(P<0.01).Further pairwise comparison showed that Th17 ranked in the order of high and low was control group>study group>healthy group,Treg in the order of high and low was healthy group>study group>control group,the levels of IL-17 and IL-22 in the order of high and low were control group>study group>healthy group,and the levels of IL-10 and TGF-β1 in the order of high and low were healthy group>study group>control group,with statistical significance(P<0.05).Conclusion Zhoufei Pingchuan Capsules can improve the lung function,arterial blood gas and symptom score of patients with lung-kidney qi deficiency syndrome in stable stage of COPD.Its mechanism may be related to regulating the balance of Th17/Treg,down-regulating the levels of Th17,IL-17 and IL-22,and up-regulating the levels of Treg,IL-10 and TGF-β1,in order to reduce airway inflammation and regulate immune homeostasis.

Comprehensive Analysis of Proteins and Their Phosphorylation in Milk-derived Exosomes From Different Species / 生物化学与生物物理进展

ObjectiveExosomes are microvesicles which could be secreted by all cell types with diameters between 30 and 150 nm. It was widely distributed in body fluids including blood, urine, and breast milk. Exosomes are considered as potential biomarkers and drug carriers by reason of containing nucleic acids, lipids, proteins and other bioactive molecules. Milk-derived exosomes have been widely used as drug delivery carriers to treat targeted diseases with a lower cost, higher biocompatibility and lower immunogenicity. Until now, there is no research about the milk-derived exosomes phosphorylation to reveal the difference of protein phosphorylation in different species of milk. To investigate the pathways and proteins with specific functions, phosphorylated proteomic analysis of milk-derived exosomes from different species is performed, and provide new ideas for exploring diversified treatments of disease. MethodsWhey and exosomes derived from bovine, porcine and caprine milk were performed for proteomics and phosphoproteomics analysis. The relationship between milk exosome proteins from different species and signaling pathways were analyzed using bioinformatics tools. ResultsA total of 4 191 global proteins, 1 640 phosphoproteins and 4 064 phosphosites were identified from 3 species of milk-derived exosomes, and the exosome proteins and phosphoproteins from different species were significantly higher than those of whey. Meanwhile, some special pathways were enriched like Fcγ-mediated phagocytosis from bovine exosomes, pathways related with neural and immune system from caprine exosomes, positive and negative regulation of multiple activities from porcine exosomes. ConclusionIn this study, the proteomic and phosphoproteomic analyses of exosomes and whey from bovine, porcine and caprine milk were carried out to reveal the difference of composition and related signaling pathways of milk exosome from different species. These results provided powerful support for the application of exosomes from different milk sources in the field of disease treatment.

A Novel Enzootic Nasal Tumor Virus Circulating in Goats from Southern China.

Enzootic nasal tumor virus (ENTV) has two types, ENTV-1 in sheep and ENTV-2 in goats, respectively. In China, the incidence of ENTV-2 related diseases has increased year by year. In this study, we reported an outbreak of ENTV-2 in a commercial goat farm in Qingyuan city, Guangdong province, southern China. A full-length genome of ENTV-2 (designated GDQY2017), with 7479 base pairs, was sequenced. Although GDQY2017 shared the highest nucleotide identity with a Chinese ENTV-2 isolate (ENTV-2CHN4, GenBank accession number KU258873), it possesses distinct genome characteristics undescribed, including a non-continuous 21-nucleotide insertion in the gag gene and a non-continuous 12-nucleotide deletion in the env gene. Notably, most of these indel nucleotide sequences were originated from a Chinese jaagsiekte sheep retrovirus (JSRV) isolate (GenBank accession number DQ838494). In the gag and env genes, GDQY2017 was phylogenetically related to those Chinese ENTV-2 isolates and a Chinese JSRV isolate (DQ838494). For GDQY2017-like viruses, more surveillance work should be made to explain their pathogenicity in goat herds. To our knowledge, this study represents the first to demonstrate the circulating pattern of ENTV-2 in Guangdong province, China, which will help to better understand the epidemiology and genetic diversity of ENTV-2.

Key findings from the WHO collaborative study on substitution therapy for opioid dependence and HIV/AIDS.

AIMS: Opioid substitution treatment has been studied extensively in industrialized countries, but there are relatively few studies in developing/transitional countries. The aim of this study was to examine the effectiveness of opioid substitution treatment (OST) in less resourced countries. DESIGN: Longitudinal cohort study. SETTING: Purposively selected OST sites in Asia (China, Indonesia, Thailand), Eastern Europe (Lithuania, Poland, Ukraine), the Middle East (Iran) and Australia. PARTICIPANTS: Seven hundred and twenty-six OST entrants. MEASUREMENTS: Participants were interviewed at treatment entry, 3 and 6 months. Standardized instruments assessed drug use, treatment history, physical and psychological health, quality of life, criminal involvement, blood-borne virus (BBV) risk behaviours and prevalence of human immunodeficiency virus (HIV) and hepatitis C. FINDINGS: Participants were predominantly male, aged in their early 30s and had attained similar levels of education. Seroprevalence rates for HIV were highest in Thailand (52%), followed by Indonesia (28%) and Iran (26%), and lowest in Australia (2.6%). Treatment retention at 6 months was uniformly high, averaging approximately 70%. All countries demonstrated significant and marked reductions in reported heroin and other illicit opioid use; HIV (and other BBV) exposure risk behaviours associated with injection drug users (IDU) and criminal activity, and demonstrated substantial improvement in their physical and mental health and general wellbeing over the course of the study. CONCLUSIONS: OST can achieve similar outcomes consistently in a culturally diverse range of settings in low- and middle-income countries to those reported widely in high-income countries. It is associated with a substantial reduction in HIV exposure risk associated with IDU across nearly all the countries. Results support the expansion of opioid substitution treatment.