Aryldiazonium Salt-Triggered [2 + 2 + 1] Heteroannulation of Indoles by an Arylhydrazone Radical-Relayed 1,5-Hydrogen Atom Transfer.

An unprecedented three-component [2 + 2 + 1] annulation cascade of indoles with aryldiazonium salts and polyhalomethanes or acetone is presented by dual hydrogen atom transfer (HAT) and C-H functionalization. By employing readily accessible aryldiazonium salts as the radical initiators and electrophiles and polyhalomethanes and acetone as the C1 units, this method unprecedentedly constructs a pyrazole ring on an indole ring skeleton through the formation of two C-N bonds and a C-C bond in a single reaction.

Molecular Characterization of Two Wamide Neuropeptide Signaling Systems in Mollusk Aplysia.

Neuropeptides with the C-terminal Wamide (Trp-NH2) are one of the last common ancestors of peptide families of eumetazoans and play various physiological roles. In this study, we sought to characterize the ancient Wamide peptides signaling systems in the marine mollusk Aplysia californica, i.e., APGWamide (APGWa) and myoinhibitory peptide (MIP)/Allatostatin B (AST-B) signaling systems. A common feature of protostome APGWa and MIP/AST-B peptides is the presence of a conserved Wamide motif in the C-terminus. Although orthologs of the APGWa and MIP signaling systems have been studied to various extents in annelids or other protostomes, no complete signaling systems have yet been characterized in mollusks. Here, through bioinformatics, molecular and cellular biology, we identified three receptors for APGWa, namely, APGWa-R1, APGWa-R2, and APGWa-R3. The EC50 values for APGWa-R1, APGWa-R2, and APGWa-R3 are 45, 2100, and 2600 nM, respectively. For the MIP signaling system, we predicted 13 forms of peptides, i.e., MIP1-13 that could be generated from the precursor identified in our study, with MIP5 (WKQMAVWa) having the largest number of copies (4 copies). Then, a complete MIP receptor (MIPR) was identified and the MIP1-13 peptides activated the MIPR in a dose-dependent manner, with EC50 values ranging from 40 to 3000 nM. Peptide analogs with alanine substitution experiments demonstrated that the Wamide motif at the C-terminus is necessary for receptor activity in both the APGWa and MIP systems. Moreover, cross-activity between the two signaling systems showed that MIP1, 4, 7, and 8 ligands could activate APGWa-R1 with a low potency (EC50 values: 2800-22,000 nM), which further supported that the APGWa and MIP signaling systems are somewhat related. In summary, our successful characterization of Aplysia APGWa and MIP signaling systems represents the first example in mollusks and provides an important basis for further functional studies in this and other protostome species. Moreover, this study may be useful for elucidating and clarifying the evolutionary relationship between the two Wamide signaling systems (i.e., APGWa and MIP systems) and their other extended neuropeptide signaling systems.

Characterization of an Aplysia vasotocin signaling system and actions of posttranslational modifications and individual residues of the ligand on receptor activity.

The vasopressin/oxytocin signaling system is present in both protostomes and deuterostomes and plays various physiological roles. Although there were reports for both vasopressin-like peptides and receptors in mollusc Lymnaea and Octopus, no precursor or receptors have been described in mollusc Aplysia. Here, through bioinformatics, molecular and cellular biology, we identified both the precursor and two receptors for Aplysia vasopressin-like peptide, which we named Aplysia vasotocin (apVT). The precursor provides evidence for the exact sequence of apVT, which is identical to conopressin G from cone snail venom, and contains 9 amino acids, with two cysteines at position 1 and 6, similar to nearly all vasopressin-like peptides. Through inositol monophosphate (IP1) accumulation assay, we demonstrated that two of the three putative receptors we cloned from Aplysia cDNA are true receptors for apVT. We named the two receptors as apVTR1 and apVTR2. We then determined the roles of post-translational modifications (PTMs) of apVT, i.e., the disulfide bond between two cysteines and the C-terminal amidation on receptor activity. Both the disulfide bond and amidation were critical for the activation of the two receptors. Cross-activity with conopressin S, annetocin from an annelid, and vertebrate oxytocin showed that although all three ligands can activate both receptors, the potency of these peptides differed depending on their residue variations from apVT. We, therefore, tested the roles of each residue through alanine substitution and found that each substitution could reduce the potency of the peptide analog, and substitution of the residues within the disulfide bond tended to have a larger impact on receptor activity than the substitution of those outside the bond. Moreover, the two receptors had different sensitivities to the PTMs and single residue substitutions. Thus, we have characterized the Aplysia vasotocin signaling system and showed how the PTMs and individual residues in the ligand contributed to receptor activity.

Recent Advances in N-O Bond Cleavage of Oximes and Hydroxylamines to Construct N-Heterocycle.

Oximes and hydroxylamines are a very important class of skeletons that not only widely exist in natural products and drug molecules, but also a class of synthon, which have been widely used in industrial production. Due to weak N-O σ bonds of oximes and hydroxylamines, they can be easily transformed into other functional groups by N-O bond cleavage. Therefore, the synthesis of N-heterocycle by using oximes and hydroxylamines as nitrogen sources has attracted wide attention. Recent advances for the synthesis of N-heterocycle through transition-metal-catalyzed and radical-mediated cyclization classified by the type of nitrogen sources and rings are summarized. In this paper, the recent advances in the N-O bond cleavage of oximes and hydroxylamines are reviewed. We hope that this review provides a new perspective on this field, and also provides a reference to develop environmentally friendly and sustainable methods.

Distribution fractions and potential ecological risk assessment of heavy metals in mangrove sediments of the Greater Bay Area.

The restoration of mangrove in coastal wetlands of China has been started since the 1990s. However, various pollutants, especially for heavy metals (HMs), contained in wastewater might present a significant risk to mangrove forests during the restoration. In this study, sediments of five typical mangrove wetlands with varying restoration years and management measures in the Greater Bay Area were collected to evaluate the distribution fractions and potential ecological risk of HMs. Cd (0.2-1.6 mg/kg) was found in high concentrations in the exchangeable fraction (37.8-71.5%), whereas Cu (54.2-94.8 mg/kg), Zn (157.6-332.6 mg/kg), Cr (57.7-113.6 mg/kg), Pb (36.5-89.9 mg/kg), and Ni (29.7-69.5 mg/kg) primarily presented in residual fraction (30.8-91.9%). According to the geo-accumulation index (Igeo) analysis, sediment Cd presented a high level of pollution (3 ≤ Igeo ≤ 4), while Zn and Cu were associated with moderately pollution (1 ≤ Igeo ≤ 2). Besides, high ecological risk of Cd was found in sediments of five mangroves, with risk assessment code (RAC) ranging from 45.9 to 84.2. Redundancy analysis revealed that the content of NO3--N was closely related to that of HMs in sediments and, pH value and NO3--N concentration affected the distribution of HMs geochemical fractions. High concentration of HMs in QA and NS sampling sites was caused by the formerly pollutants discharge, resulting in these sediments still with a higher HM pollution level after the plant of mangrove for a long period. Fortunately, strict drainage standards for industrial activities in Shenzhen significantly availed for decreasing HMs contents in mangrove sediments. Therefore, future works on mangrove conversion and restoration should be linked to the water purification in the GBA.

Preoperative and postoperative clinical signatures of postgastrectomy venous thromboembolism in patients with gastric cancer: A retrospective cohort study.

BACKGROUND: This study aimed to identify preoperative and postoperative risk factors of venous thromboembolism (VTE) after gastrectomy in gastric cancer (GC) patients. METHODS: 757 GC patients underwent gastrectomy at our institution and 246 patients with elevated postoperative D-dimer levels who received Doppler ultrasonography of lower/upper extremity veins were enrolled. Clinicopathological factors data were collected, and the differences in clinicopathological factors between postoperative VTE (+) and VTE (-) groups were analyzed. Univariate and multivariate logistic regression analyses were performed to identify independent risk factors of postgastrectomy VTE. RESULTS: Of 246 patients with elevated postgastrectomy D-dimer concentrations, 74 patients showed thrombosis in lower/upper extremity veins. Among preoperative factors, age, WBC level, D-dimer concentration, and blood glucose level were significantly higher in the postoperative VTE (+) group. Among the postoperative factors, hemoglobin level was significantly lower in the postoperative VTE (+) group. Among the pathological factors, tumor stage, depth of invasion and TNM classification indicated higher malignancy in the postoperative VTE (+) group. Univariate logistic regression analysis indicated age, preoperative blood glucose level, postoperative hemoglobin level, tumor stage, depth of invasion, and TNM classification as the independent risk factors for postgastrectomy VTE, whereas multivariate logistic regression analysis revealed age and tumor stage as independent risk factors for postgastrectomy VTE. CONCLUSION: Our study revealed that age, preoperative blood glucose level, postoperative anemia, and tumor malignancy were independent risk factors for GC patients exhibiting postgastrectomy VTE. Therefore, the perioperative monitoring, assessment and management of risk factors are important in achieving better outcomes after gastrectomy.

AI protein structure prediction-based modeling and mutagenesis of a protostome receptor and peptide ligands reveal key residues for their interaction.

The protostome leucokinin (LK) signaling system, including LK peptides and their G protein-coupled receptors, has been characterized in several species. Despite the progress, molecular mechanisms governing LK peptide-receptor interactions remain to be elucidated. Previously, we identified a precursor protein for Aplysia leucokinin-like peptides (ALKs) that contains the greatest number of amidated peptides among LK precursors in all species identified so far. Here, we identified the first ALK receptor from Aplysia, ALKR. We used cell-based IP1 activation assays to demonstrate that two ALK peptides with the most copies, ALK1 and ALK2, activated ALKR with high potencies. Other endogenous ALK-derived peptides bearing the FXXWX-amide motif also activated ALKR to various degrees. Our examination of cross-species activity of ALKs with the Anopheles LK receptor was consistent with a critical role for the FXXWX-amide motif in receptor activity. Furthermore, we showed, through alanine substitution of ALK1, the highly conserved phenylalanine (F), tryptophan (W), and C-terminal amidation were each essential for receptor activation. Finally, we used an artificial intelligence-based protein structure prediction server (Robetta) and Autodock Vina to predict the ligand-bound conformation of ALKR. Our model predicted several interactions (i.e., hydrophobic interactions, hydrogen bonds, and amide-pi stacking) between ALK peptides and ALKR, and several of our substitution and mutagenesis experiments were consistent with the predicted model. In conclusion, our results provide important information defining possible interactions between ALK peptides and their receptors. The workflow utilized here may be useful for studying other ligand-receptor interactions for a neuropeptide signaling system, particularly in protostomes.

Identification of an allatostatin C signaling system in mollusc Aplysia.

Neuropeptides, as pervasive intercellular signaling molecules in the CNS, modulate a variety of behavioral systems in both protostomes and deuterostomes. Allatostatins are neuropeptides in arthropods that inhibit the biosynthesis of juvenile hormones. Based on amino acid sequences, they are divided into three different types in arthropods: allatostatin A, allatostatin B, allatostatin C. Allatostatin C (AstC) was first isolated from Manduca sexta, and it has an important conserved feature of a disulfide bridge formed by two cysteine residues. Moreover, AstC appears to be the ortholog of mammalian somatostatin, and it has functions in common with somatostatin, such as modulating feeding behaviors. The AstC signaling system has been widely studied in arthropods, but minimally studied in molluscs. In this study, we seek to identify the AstC signaling system in the marine mollusc Aplysia californica. We cloned the AstC precursor from the cDNA of Aplysia. We predicted a 15-amino acid peptide with a disulfide bridge, i.e., AstC, using NeuroPred. We then cloned two putative allatostatin C-like receptors and through NCBI Conserved Domain Search we found that they belonged to the G protein-coupled receptor (GPCR) family. In addition, using an inositol monophosphate 1 (IP1) accumulation assay, we showed that Aplysia AstC could activate one of the putative receptors, i.e., the AstC-R, at the lowest EC50, and AstC without the disulfide bridge (AstC') activated AstC-R with the highest EC50. Moreover, four molluscan AstCs with variations of sequences from Aplysia AstC but with the disulfide bridge activated AstC-R at intermediate EC50. In summary, our successful identification of the Aplysia AstC precursor and its receptor (AstC-R) represents the first example in molluscs, and provides an important basis for further studies of the AstC signaling system in Aplysia and other molluscs.

Comparative evaluation of rotational stability of toric IOLs with four-eyelet vs two-eyelet capsular tension rings in eyes with high myopia.

AIM: To compare the rotational stability of Toric intraocular lens (IOLs) implantation combined with four-eyelet or two-eyelet capsular tension rings (CTRs) in eyes with high myopia and cataract. METHODS: This prospective randomized controlled interventional study included 33 eyes which had preoperative corneal astigmatism ≥1.5 D and ocular axial length ≥25.5 mm. These eyes were randomly divided into two groups to undergo phacoemulsification and toric IOL implantation with either four-eyelet CTR implantation (group A, n=16) or two-eyelet CTR implantation (group B, n=17). Uncorrected visual acuity (UCVA), best-corrected visual acuity (BCVA), phoropter examination results, and toric IOL rotation degrees were tested 6mo after the surgery. RESULTS: In both groups, the toric IOL was in the capsular sac 6mo after surgery. The difference between the two groups in terms of visual outcome was not found to be statistically significant (P>0.05) at a follow-up of 6mo. The mean residual astigmatism values were 0.56±0.22 D and 0.92±0.24 D in A and B groups, respectively (P<0.001). The mean rotation degree of IOL was 1.00°±0.73° in group A and 3.53°±1.46° in group B (P<0.001). CONCLUSION: In cataract patients with high myopia and astigmatism, four-eyelet CTR can effectively increase the rotation stability of toric IOLs, achieving the desired goal of correcting corneal astigmatism.

[Effects of consecutive application of composted sewage sludge on active organic nitrogen fractions in the sandy fluvo-aquic soil.] / è¿žç»­æ–½ç”¨æ— å®³åŒ–æ±¡æ³¥å †è‚¥å¯¹æ²™è´¨æ½®åœŸæ´»æ€§æœ‰æœºæ°®ç»„åˆ†çš„å½±å“.

A field experiment was conducted with sandy fluvo-aquic soil under wheat-maize rotation system during 2012 to 2016 in Henan Province, with four treatments: Chemical fertilizer (CK), composted sewage sludge (CS) 15 t·hm-2(CS1), 30 t·hm-2(CS2), 45 t·hm-2(CS3). We investigated the effects of CS on soil active organic nitrogen and its allocation ratio. Results showed that CS significantly enhanced soil water content, decreased soil pH, increased contents of soil organic carbon and total nitrogen (TN) and available nutrients. Soil TN was increased by 93.1%-284.3%. The CS3 treatment significantly improved the content of light fraction organic nitrogen (LFON), particulate organic nitrogen (PON) and microbial biomass nitrogen (MBN). CS treatments promoted the allocation of PON/TN and MBN/TN. with an enhancement of 12.3% and 539.9% in the CS2 treatment, respectively. The distributions of LFON/TN and DON/TN were decreased by 17.3%-40.1% and 38.5%-71.3%, respectively. The contribution of soil active organic nitrogen fractions to TN was: PON>LFON>DON>MBN. Results from the principal component analysis showed that organic nitrogen and their distribution had high load value in CS2 and CS3 treatments. Results from redundancy analysis showed that positive correlations existed between the soil physicochemistry, enzymatic activity and soil active organic nitrogen as well as their distribution. In summary, CS had positive impacts on soil organic matter, soil properties, and the formation of active organic nitrogen. The rates of 30 and 45 t·hm-2 CS application had significant effects on soil fertility, which could be an effective way to improve sandy fluvo-aquic soil quality.

Traditional Chinese medicine Qiliqiangxin attenuates phenylephrine-induced cardiac hypertrophy via upregulating PPARγ and PGC-1α.

BACKGROUND: Clinical study has demonstrated that the traditional Chinese medicine Qiliqiangxin (QLQX) has protective effects on heart failure. Phenylephrine (PE) is an important inducing factor for cardiac hypertrophy and our previous studies have showed that QLQX attenuates PE-induced cardiac hypertrophy. Besides, QLQX protects against cardiac remodeling after myocardial infarction via activating PPARγ. However, whether QLQX prevents PE-induced cardiac hypertrophy through PPARγ and its coactivator PGC-1α is still unknown. METHODS: The effects of QLQX were investigated based on PE induced cardiac hypertrophy mouse models. Echocardiography and hematoxylin-eosin (HE) staining were used to determine cardiac function and cross-sectional area, respectively. Quantitative real time PCR (qRT-PCR) was used to determine ANP and BNP expressions. Based on primary neonatal rat ventricular cardiomyocytes (NRVMs) treated with PE, the cell size and expressions of ANP and BNP were determined by immunofluorescent staining and qRT-PCR, respectively. In addition, western blot was used to determine PPARγ and PGC-1α expressions. RESULTS: In present study, we confirmed that QLQX could significantly attenuate cardiac hypertrophy in mice treated with PE. Then we showed that PPARγ and PGC-1α were downregulated in PE-induced cardiac hypertrophy, and QLQX could block the decrease of PPARγ and PGC-1α both in vitro and in vivo. Importantly, we found that PPARγ inhibitors or PGC-1α siRNAs eliminated the protective effects of QLQX on PE-induced cardiac hypertrophy. CONCLUSIONS: Our study suggested that QLQX prevents from PE-induced cardiac hypertrophy by activating PPARγ and its coactivator PGC-1α.

[Influence of the Application of Non-Hazardous Sewage Sludge on the Evolution of Soil Carbon Pool and Carbon Pool Management Index].

The products of non-hazardous commercial sewage sludge were utilized in the accordance of the national standard in the experiment with sandy fluvo-aquic soil under wheat-maize rotation system during 2013 to 2015. The experiment focused on the effects of the use of non-hazardous sewage sludge on soil carbon pool and carbon pool management index to provide theoretical and technical basis for the resource utilization of non-hazardous sewage sludge. The results showed that compared with CK, soil amended sludge significantly improved soil total organic carbon (TOC), soil microbial biomass carbon (SMBC), labile organic carbon (LOC),and dissolved organic carbon (DOC) to 8.40-14.74 g·kg-1, 164.45-257.45 mg·kg-1, 3.58-4.88 g·kg-1and 81.16-101.58 mg·kg-1, soil amended sludge significantly enhanced SMBC, LOC and DOC by 84.00%-188.07%, 26.26%-58.03%, and 109.58%-185.39% (P<0.05) respectively, and 45 t·hm-2 sewage sludge (W3) had the most significant impact on soil carbon pool. The soil microbial entropy (SMBC/TOC), and utilization of labile organic carbon (LOC/TOC) increased by 8.02%-2.77% and 13.75%-46.48% respectively, and the utilization of dissolved organic carbon (DOC/TOC) significantly decreased by 153.45%-195.40% (P<0.05). SMBC/TOC, LOC/TOC, and DOC/TOC declined in treatments of soil amended sludge due to increased application of sewage sludge, which indicated that soil amended with 45 t·hm-2 of the sewage sludge improved the content of stable carbon resulting in the decease of the ratio. L and LI decreased with the increasing amount of sewage sludge indicating that soil amended with 45 t·hm-2 of the sewage sludge improved the content of stable carbon and was conducive to the accumulation of organic carbon. Soil amended with sludge significantly increased carbon pool management index (CMPI) by 153.45%-195.40% (P<0.05), and W3 had the most significant effect on CMPI. Compared with TOC, CMPI could be more sensitive and direct to reflect the dynamic changes of soil nutrients and carbon pools through the correlation analysis and redundancy analysis. In summary, the application of 15-45 t·hm-2 sewage sludge could significantly enhance the soil carbon pool and carbon pool management index, especially 45 t·hm-2 sewage sludge (W3).

[Effects of Different Organic Materials on Bio-availability of Cd, Pb in a Contaminated Greenhouse Soil].

A two-season soil culture experiment was continually conducted to study the effects of three organic materials,namely Danty, Peat and Biochar,respectively,on combined forms and bio-availability of cadmium (Cd) and lead (Pb) under Cd and Pb in single and combined pollution during 2014-2015. This study aimed to provide a theoretical basis for remediating greenhouse soil contaminated with heavy metals Cd and Pb using organic materials. The main results were as follows: the content of DTPA-Cd decreased significantly by 11.9%(P<0.05) in the soil of Cd and Pb combined pollution with the application of danty, the total of exchangeable Cd, carbonate-bound Cd and iron-manganese oxide bound Cd was reduced by 26.4% and 34.4% in the soil, and the accumulation of Cd in shoots was significantly cut down by 32.0% and 28.0% (P<0.05) respectively in Cd single and Cd and Pb combined pollution after applying danty; the content of DTPA-Pb was significantly depleted by 83.5% and 83.8%(P<0.05), the total of exchangeable Pb, carbonate-bound Pb and iron-manganese oxide bound Pb was decreased by 34.5% and 13.9% in the soil and the accumulation of Pb in shoots was significantly reduced by 32.0% and 30.0% (P<0.05) in Pb single and Cd and Pb combined pollution respectively as the use of danty. The content of DTPA-Cd was significantly decreased by 18.9% (P<0.05) in the soil of Cd and Pb combined pollution with the use of peat, the accumulation of Cd in shoots was reduced by 38.0% and 23.4%(P<0.05) in Cd single and Cd-Pb combined pollution respectively after the application of peat; The content of DTPA-Pb was significantly decreased by 2.7% and 7.2%(P<0.05), the total of exchangeable Pb, carbonate-bound Pb and iron-manganese oxide bound Pb was decreased by 15.8% and 14.6% in the soil and the accumulation of Pb in shoots was significantly reduced by 12.7% and 23.4% (P<0.05) respectively in Pb single and Cd and Pb combined pollution due to the application of peat. The pH value of the soil was increased by the use of biochar, the content of DTPA-Cd was reduced by 4.7% and 15.0% respectively in the soil of Cd single and Cd and Pb combined pollution, and the accumulation of Cd in shoots was significantly minified by 38.0% and 23.4% respectively in Cd single and Cd and Pb combined pollution as the application of biochar. The content of DTPA-Pb was decreased by 6.8% and 1.0% and the total of exchangeable Pb, carbonate-bound Pb and iron-manganese oxide bound Pb was cut down by 11.9% and 30.0% in the soil of Pb single and Cd and Pb combined pollution respectively by the application of biochar. The content of Cd in plant was most significantly positively correlated with the content of exchangeable Cd in soil (P<0.01). The content of Pb in plant was most significantly positively correlated with the total of exchangeable Pb, carbonate-bound Pb and iron-manganese oxide bound Pb (P<0.01). In summary, after the application of danty, peat and biochar, the proportion of highly active content of Cd and Pb was minified through adjusting soil pH and immobilizing Cd and Pb by means of adsorption and complexation in greenhouse soil. With the analysis of input-output, the application of danty, peat and biochar costs 4050 yuan·hm-2, 11250 yuan·hm-2 and 22500 yuan·hm-2, respectively. Thus danty, peat and biochar could be used as effective organic agents in the remediation of Cd and Pb contaminated greenhouse soil.

[Pretreatment technology for fly ash from MSWI and the corresponding study of chloride behavior].

The introduced pretreatment technology, WCCB (Washing + Calcination), was effective to reduce chlorides in fly ash by consuming relatively low energy for recycling fly ash as the raw material for cement industry. The washing conditions are: twice-washing, liquid/solid = 3, mixing speed = 150 r x min(-1), 1st mixing time = 5 min, and 2nd mixing time = 10 min. The original incinerator was used for the calcination process, 1 000 degrees C, 10% O2 and dwelling time of 1 hour were adopted. By adopting X-ray absorption near edge structure and X-ray diffraction, the behavior of chlorides was explained and NaCl, KCl and CaCl2 are the main form of chlorides existing in fly ash. The reagent used in the air pollution control system to neutralize the acid component in the discharged gas surely acted a very important role in the formation of chlorides. The insoluble chlorides in fly ash had a very similar structure as that of Friedel's salt, which was related with CaCl2.

Role for granulocyte colony stimulating factor in angiotensin II-induced neutrophil recruitment and cardiac fibrosis in mice.

BACKGROUND: Granulocyte colony stimulating factor (G-CSF) is a key mediator of neutrophil infiltration and is profibrotic in the liver, lung, and infarcted heart, but its roles in angiotensin II (Ang II)-induced hypertension and cardiac remodeling have not been fully determined. Thus, we sought to investigate the causal relation of G-CSF to neutrophil recruitment and cardiac fibrosis in C57BL/6J mice. METHODS: Hypertension and cardiac fibrosis were induced in wild-type (WT) mice receiving continuous infusion of Ang II (1,500ng/kg/min). After 7 days, heart sections were stained with hematoxylin and eosin, Masson's trichrome, and immunohistochemistry. The mRNA expression of cytokines was detected by real-time polymerase chain reaction analysis. The protein levels were measured by Western blot analysis. RESULTS: After Ang II infusion, myocardial G-CSF expression was significantly elevated in the hearts. Moreover, WT mice exhibited increased blood pressure, marked neutrophil accumulation, proinflammatory cytokine expression, reactive oxygen species production, and cardiac fibrosis after 7 days of Ang II infusion. However, administration of anti-G-CSF neutralizing antibody, but not with control immunoglobulin G, significantly attenuated these effects. In addition, neutralizing G-CSF antibody reversed Ang II-induced activation of ERK1/2, STAT3, and AKT signaling pathways in the hearts. CONCLUSIONS: This study demonstrates that G-CSF plays a critical role in hypertension and cardiac fibrosis and targeting this cytokine may be a novel therapeutic strategy to ameliorate hypertensive heart disease.

Ginsenoside Re reduces insulin resistance through activation of PPAR-γ pathway and inhibition of TNF-α production.

ETHNOPHARMACOLOGICAL RELEVANCE: Panax ginseng is a well-known traditional Chinese medicine and has been used for treatment of various diseases for more than four thousand years in Asia. Ginseng saponins or ginsenosides, the active constituents are reported to possess antidiabetic activity, but their antihyperglycemic mechanisms are not fully elucidated. In the present study, the mechanisms of action of ginsenoside Re were investigated in vitro models. MATERIALS AND METHODS: 3T3-L1 cells were chosen as the model to investigate the molecular mechanisms of action of ginsenoside Re. Influence of ginsenoside Re on the adipogenesis was examined by determining TG levels in 3T3-L1 adipocytes by the method of TG oxidation enzyme. Glucose uptake in 3T3-L1 cells stimulated by insulin in the absence or presence of ginsenoside Re were quantified by measuring (3)H-2-deoxy-d-glucose levels. Cytokine proteins released into the medium including adiponectin and TNF-α were tested using respective ELISA kits. In addition, real time RT-PCR was conducted to investigate the expression changes of PPAR-γ and its responsive genes, ap2, adiponectin, IRS-1, GLUT4 and TNF-α. And western blot analysis was performed to determine the translocation of GLUT4. Finally, effects of ginsenoside Re on NO production in 3T3-L1 adipocytes and in macrophages were investigated through measurement of nitrite concentration by Griess reagent. RESULTS: Ginsenoside Re induced adipogenesis of 3T3-L1 adipocytes by accumulating TG, increased glucose uptake and up-regulated PPAR-γ2, IRS-1, ap2 and adiponectin genes expressions. Meanwhile, Re also increased production and release of adiponectin. Although having no effects on GLUT4 gene expression, Re facilitated GLUT4 protein translocation to the membranes. In addition, Re inhibited the expression and release of TNF-α. Finally, Re did not show inhibitory effects on NO production both in 3T3-L1 cells stimulated by LPS, TNF-α and IFN-γ and in LPS-stimulated mouse peritoneal macrophages. CONCLUSIONS: Ginsenoside Re exhibited the action of reducing insulin resistance through activation of PPAR-γ pathway by directly increasing the expressions of PPAR-γ2 and its responsive genes, adiponectin, IRS-1, ap2, inhibiting TNF-α production and facilitating the translocation of GLUT4 to promote glucose uptake and disposal in 3T3-L1 adipocytes.