Dissolving Chitin by Novel Deep Eutectic Solvents for Effectively Enzymatic Hydrolysis.

Chitin is the most productive nitrogen-containing polysaccharide in nature with immense potential for transforming into a range of chemicals. However, its dense crystal structure poses a challenge for depolymerization, limiting its applications. To overcome these challenges, a novel series of deep eutectic solvents (DESs) based on benzyltrimethylammonium chloride (TMBAC) as the hydrogen bond acceptor was developed. These TMBAC-based DESs, in combination with lactic acid, oxalic acid, and malic acid as the hydrogen bond donor demonstrated efficient chitin dissolution, achieving a solubility of up to 12% and an 88% recovery rate of regenerated chitin. The regenerated chitin was characterized using XRD, FT-IR, SEM, and 13C CP-MAS NMR, which indicated the preservation of chitin's chemical structure, a significant decrease in crystallinity, and a reduction in the molecular weight. Furthermore, the enzymatic hydrolysis efficiency of chitin was nearly doubled after treatment with TMBAC-based DESs, surpassing the effectiveness of untreated chitin. This approach holds promise for facilitating subsequent transformation and utilization of chitin.

The emerging role of lncRNAs in osteoarthritis development and potential therapy.

Osteoarthritis impairs the functions of various joints, such as knees, hips, hands and spine, which causes pain, swelling, stiffness and reduced mobility in joints. Multiple factors, including age, joint injuries, obesity, and mechanical stress, could contribute to osteoarthritis development and progression. Evidence has demonstrated that genetics and epigenetics play a critical role in osteoarthritis initiation and progression. Noncoding RNAs (ncRNAs) have been revealed to participate in osteoarthritis development. In this review, we describe the pivotal functions and molecular mechanisms of numerous lncRNAs in osteoarthritis progression. We mention that long noncoding RNAs (lncRNAs) could be biomarkers for osteoarthritis diagnosis, prognosis and therapeutic targets. Moreover, we highlight the several compounds that alleviate osteoarthritis progression in part via targeting lncRNAs. Furthermore, we provide the future perspectives regarding the potential application of lncRNAs in diagnosis, treatment and prognosis of osteoarthritis.

Identification and Characterization of a Novel Mannanase from Klebsiella grimontii.

Konjac glucomannan (KGM) is a natural polysaccharide derived from konjac, which has been widely used in various fields due to its numerous beneficial properties. However, the high viscosity and water absorption of KGM limit its application. Compared with KGM, Konjac glucomannan oligosaccharides (KGMOS) have higher water solubility and stronger application value. In this paper, a novel mannanase KgManA was cloned from Klebsiella grimontii to develop a new KGMOS-producing enzyme. Bioinformatic analysis shows that the structural similarity between KgManA and other enzymes was less than 18.33%. Phylogenetic analysis shows that KgManA shares different branches with the traditional mannanases containing the CMB35 domain, indicating that it is a novel mannanase. Then, the enzymatic properties were determined and substrate specificity was characterized. Surprisingly, KgManA is stable in a very wide pH range of 3.0 to 10.0; it has a special substrate specificity and seems to be active only for mannans without galactose in the side chain. Additionally, the three-dimensional structure of the enzyme was simulated and molecular docking of the mannotetraose substrate was performed. As far as we know, this is the first report to characterize the enzymatic properties and to simulate the structure of mannanase from K. grimontii. This work will contribute to the development and characterization of novel K. grimontii-derived mannanases. The above results indicate that KgManA is a promising tool for the production of KGMOS.

First Report of Neopestalotiopsis clavispora Causing Postharvest Fruit Rot on Actinidia arguta in Liaoning Province, China.

Actinidia arguta, commonly called hardy kiwifruit or kiwiberry, is a perennial vine of Actinidiaceae Actinidia genus. Understanding the main pathogens that cause the fruit rot of A. arguta during storage is of great significance for finding strategies to prevent fruit rot. In September 2020, the A. arguta (Sieb.et Zucc.) Planch. ex Miq. LD133 was harvested from a farm in Dandong City, Liaoning Province, China (40°31'N, 124°20'E). After being stored at room temperature for about a week, the fruit rotted (no mechanical damage or wound). Initial symptoms were localized irregular spots, which then became soft and the spots connected into large, flaky, light brown lesions. Later symptoms were dark brown lesions and rot that affect the entire fruit. The strain that shows the same morphology as observed in the four decaying tissues was isolated on PDA. The strain was white, edge irregular and surface wavy, and the reverse side was pale yellow (Fig. S1A). The black viscous acervuli appeared on the surface of the mycelium after 7 to 10 days at 25℃ with 12 hours photoperiod. Conidia were fusiform to ellipsoid, straight to slightly curved, 5.7×25.9 µm (width × length), n=30, with five versicolor cells (three brown median cells, two hyaline cells on apical and basal). The apical cell generally contains one to four appendages on conidia (Fig. S1B). For identify the selected strains, three genetic regions (ITS, TUB and TEF 1-α) were used for amplification and sequencing. These sequences of pathogen shared 98 to 100% homology with Neopestalotiopsis clavispora. Then, a phylogenetic tree was constructed by the Bayesian algorithm using PhyloSuite (v1.2.2) (Zhang et al. 2020). Based on the morphological and molecular characterization, the pathogen was identified as Neopestalotiopsis clavispora (Chamorro et al. 2016). Next, pathogenicity of the screened strains was determined by wound inoculation method. Ripe healthy fruits are immersed in 1% NaClO, rinsed twice with sterile water. A sterile needle was used to penetrate 1-2mm of peel, and then inoculate hyphae (about 5mm in length, 1mm in diameter), and 10 µL of sterile water as a control. The treated fruits are stored in an artificial climate chamber (22°C, 70% relative humidity with 12 hours photoperiod). The inoculated fruit began to show signs of rot on the second day, and after 6 days, lesions similar to those found during storage of the pathogenic isolated fruit appeared. Similarly, 10 µL (106 conidia/mL) conidial suspension exhibited decay symptoms. Compared with conidial suspension as inoculum, hyphae has shorter incubation period and stronger pathogenicity to fruit (Fig. S2). The pathogen was re-isolated from these infected fruits and identified as N. clavispora, thus fulfilling Koch's postulates. N. clavispora has been reported causing root and crown rot on strawberry in Spain, Argentina, Uruguay and Italy (Chamorro et al. 2016; Gilardi et al. 2019; Machin et al. 2019; Obregon et al. 2018), and as a pathogen on blueberry in Spain and Korea (Borrero et al. 2018; Lee et al. 2019), and causing leaf spot on macadamia in Brazil (Santos et al. 2019), and causing leaf spot on Syzygium cumini in India (Banerjee and Rana 2020). However, to our knowledge, this is the first report of Neopestalotiopsis clavispora causing postharvest fruit rot of A. argute in the worldwide. The identification of the pathogen is of great significance for conducting research on A. argute fruit preservation to prolong its shelf life and improve its merchantability.

Identification and characterization of a novel glucomannanase from Paenibacillus polymyxa.

Konjac glucomannan oligosaccharide has attracted much attention due to its broad biological activities. Specific glucomannan degrading enzymes are effective tools for the production of oligosaccharides from konjac glucomannan. However, there are still few reports of commercial enzymes that can specifically degrade konjac glucomannan. The gene ppgluB encoding a glucomannanase consisting of 553 amino acids (61.5 kDa) from Paenibacillus polymyxa 3-3 was cloned and heterologous expressed in Escherichia coli BL21 (DE3). The recombinant PpGluB showed high specificity for the degradation of konjac glucomannan. Moreover, the hydrolytic products of PpGluB degrade konjac glucomannan were a series of oligosaccharides with degrees of polymerisation of 2-12. Furthermore, the biochemical properties indicated that PpGluB is the optimal active at 45 to 55 °C and pH 5.0-6.0, and shows highly pH stability over a very broad pH range. The present characteristics indicated that PpGluB is a potential tool to be used to produce oligosaccharides from konjac glucomannan.

Metal-Free Trifluoroalkylation of Quinoxalin-2(1H)-ones with Unactivated Alkenes and Langlois' Reagent.

A K2S2O8-mediated three-component protocol has been developed for the construction of 3-trifluoroalkylated quinoxalin-2(1H)-ones under metal-free conditions. The present reaction could be accomplished through the trifluoroalkylation of quinoxalin-2(1H)-ones with unactivated alkenes and Langlois' reagent (CF3SO2Na), which provided a highly attractive approach to access a series of biologically important 3-trifluoroalkylated quinoxalin-2(1H)-ones.

A Novel 2,5-Furandicarboxylic Acid Biosynthesis Route from Biomass-Derived 5-Hydroxymethylfurfural Based on the Consecutive Enzyme Reactions.

2,5-Furandicarboxylic acid (FDCA) is a promising bio-based building block as a green alternative to petroleum-based terephthalate in polymer production. Most of FDCA is produced by the oxidation of 5-hydroxymethylfurfural (HMF), which is derived from hexose. Although the chemical conversion is widely applied, the biocatalytic conversion is expected due to the relatively mild condition and fewer toxic chemicals consumption. However, it's difficult to catalyze the conversion of HMF to FDCA by a single enzyme. Here, a newly enzymatic cascade reaction process was introduced with a yield of 94.0% by the combination of 5-hydroxymethylfurfural oxidase (HMFO) and lipase. Briefly, a flavine adenosine dinucleotide independent (FAD-independent) HMFO of Methylovorus sp. MP688 was used to convert HMF to 2,5-diformylfuran (DFF) and 5-formylfuroic acid (FFA), which consecutively transformed to FDCA by a lipase Novozym 435. To facilitate the purification, a coupled alkali precipitation was developed to recover FDCA from organic solvent with an improved purity from 84.4 to 99.0% and recovery of 78.1%. This work will help to construct the green biorefinery route for the bulk FDCA from biomass by enzymes.

Identification and characterization of thermostable endo-polygalacturonase II B from Aspergillus luchuensis.

Endo-polygalacturonase II B (PgaB) from Aspergillus luchuensis was orthologous to endo-polygalacturonase from Aspergillus niger with mutant sites Thr42Ser and Glu52Ala. Mature pgaB gene was cloned from the genomic DNA of A. luchuensis and secreted expressed with over 90% purity in Pichia Pastoris and reached 1.0 g/L after 144 hr culture. The recombinant PgaB was further purified by Ni-NTA chromatography. Using polygalacturonic acid (PGA) as substrate, the optimal condition for PgaB activity was 40°C and pH 4.5, respectively. Km and Vmax of PgaB were 0.19 mmol/l and 103.58 µmol min-1  mg-1 , respectively. The relative activity of PgaB remained more than 60% and 40% of maximum activity at 50 and 60°C for 7 hr. PgaB increased the light transmittance by 85% and showed high efficiency in juice clarification. The main product was galacturonic acid oligosaccharides with degrees of polymers (DP) 1-3. The PgaB is a potential pectinolytic enzyme in food industries. PRACTICAL APPLICATIONS: Endo-polygalacturonase II B (PgaB) was identified from Aspergillus luchuensis, a filamentous fungus widely used in food and beverage fermentation in East Asia. PgaB still kept its most activity at 60°C for 7 hr. Polygalacturonic acid (PGA) can be digested effectively by the PgaB and the main products are galacturonic acid oligosaccharides with degrees of polymers (DP) 1-3. PgaB shows high efficiency in juice clarification. The PgaB is a potential pectinolytic enzyme for the applications in food industries.

Production and structural characterization of a new type of polysaccharide from nitrogen-limited Arthrospira platensis cultivated in outdoor industrial-scale open raceway ponds.

BACKGROUND: Carbohydrates are major biomass source in fuel-targeted biorefinery. Arthrospira platensis is the largest commercialized microalgae with good environmental tolerance and high biomass production. However, the traditional target of A. platensis cultivation is the protein, which is the downstream product of carbohydrates. Aiming to provide the alternative non-food carbohydrates source, the feasible manipulation technology on the cultivation is needed, as well as new separation methodology to achieve maximum utilization of overall biomass. RESULTS: The present study aimed to demonstrate the feasibility of industrially producing carbohydrate-enriched A. platensis and characterize the structure of the polysaccharide involved. Cultivated in industrial-scale outdoor open raceway ponds under nitrogen limitation, A. platensis accumulated maximally 64.3%DW of carbohydrate. The maximum biomass and carbohydrate productivity reached 27.5 g m-2 day-1 and 26.2 g m-2 day-1, respectively. The efficient extraction and purification of the polysaccharides include a high-pressure homogenization-assisted hot water extraction followed by flocculation with a non-toxic flocculant ZTC1 + 1, with the polysaccharide purity and total recovery reaching 81% and 75%, respectively. The purified polysaccharide was mainly composed of (1→3)(1→4)- or (1→3)(1→2)-α-glucan with a molecular weight of 300-700 kDa, which differed from the commonly acknowledged glycogen. CONCLUSIONS: By the way of controlled nitrogen limitation, the high carbohydrate production of A. platensis in the industrial scale was achieved. The α-glucan from A. platensis could be a potential glucose source for industrial applications. A non-toxic separation method of carbohydrate was applied to maintain the possibility of utilization of residue in high-value field.

Pectin Oligosaccharides Ameliorate Colon Cancer by Regulating Oxidative Stress- and Inflammation-Activated Signaling Pathways.

Colon cancer (CC) is the third common neoplasm worldwide, and it is still a big challenge for exploring new effective medicine for treating CC. Natural product promoting human health has become a hot topic and attracted many researchers recently. Pectin, a complex polysaccharide in plant cell wall, mainly consists of four major types of polysaccharides: homogalacturonan, xylogalacturonan, rhamnogalacturonan I and II, all of which can be degraded into various pectin oligosaccharides (POS) and may provide abundant resource for exploring potential anticancer drugs. POS have been regarded as a novel class of potential functional food with multiple health-promoting properties. POS have antibacterial activities against some aggressive and recurrent bacterial infection and exert beneficial immunomodulation for controlling CC risk. However, the molecular functional role of POS in the prevention of CC risk and progression remains doubtful. The review focuses on antioxidant and anti-inflammatory roles of POS for promoting human health by regulating some potential oxidative and inflammation-activated pathways, such as ATP-activated protein kinase (AMPK), nuclear factor erythroid-2-related factor-2 (Nrf2), and nuclear factor-κB (NF-κB) pathways. The activation of these signaling pathways increases the antioxidant and antiinflammatory activities, which will result in the apoptosis of CC cells or in the prevention of CC risk and progression. Thus, POS may inhibit CC development by affecting antioxidant and antiinflammatory signaling pathways AMPK, Nrf2, and NF-κB. However, POS also can activate signal transduction and transcriptional activator 1 and 3 signaling pathway, which will reduce antioxidant and anti-inflammatory properties and promote CC progression. Specific structural and structurally modified POS may be associated with their functions and should be deeply explored in the future. The present review paper lacks the important information for the linkage between the specific structure of POS and its function. To further explore the effects of prebiotic potential of POS and their derivatives on human immunomodulation in the prevention of CC, the specific POS with a certain degree of polymerization or purified polymers are highly demanded to be performed in clinical practice.

The inhibition of LPS-induced inflammation in RAW264.7 macrophages via the PI3K/Akt pathway by highly N-acetylated chitooligosaccharide.

Chitooligosaccharide (COS) has been shown to regulate many biological functions, such as antimicrobial effect and antitumor activity. In the present study, highly N-acetylated chitooligosaccharide (NACOS) was prepared by N-acetylation of COS, and the anti-inflammatory activity of NACOS in macrophages were evaluated. The results indicated NACOS significantly suppressed the LPS-induced pro-inflammatory cytokines interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) expression. Furthermore, the increased levels of reactive oxygen species (ROS) and nitric oxide (NO) were repressed by NACOS in a dose dependent manner. However, NACOS itself had no significant effect on the cell viability and cellular morphology. Signal transduction studies demonstrated that NACOS remarkably inhibited LPS-enhanced phosphorylation of phosphatidylinositol 3-kinase (PI3K) and Akt. These findings provide a possible molecular mechanism by which NACOS inhibit LPS-induced inflammatory response in macrophages, and a basis for utilizing NACOS in pharmaceutical therapy against inflammation.

Pedobacter hainanensis sp. nov., isolated from seaside soil.

A Gram-negative, aerobic, rod-shaped, motile, non-spore-forming bacterial strain, designated 13-Q(T), was isolated from seaside soil under the stacks of the red algae in Hainan province in China. Identification was carried out on the basis of polyphasic taxonomy. Phylogenetic analysis of 16S rRNA gene sequences showed that strain 13-Q(T) belonged to the genus Pedobacter, and the highest similarity was 94.4 % with Pedobacter terricola KCTC 12876(T). Strain 13-Q(T) was able to grow at 10-40 °C, in pH 5.0-10.0, in the presence of 0-2.0 % NaCl. The major fatty acids were iso-C(15:0) (40.4 %), summed feature 3 (comprising iso-C(15:0) 2-OH and/or C(16:1) ω7c) (18.9 %) and iso-C(17:0) 3-OH (18.4 %). The predominant menaquinone was MK-7. The G+C content of the genomic DNA was 42.7 mol%. Strain 13-Q(T) could be distinguished from the nearest phylogenetic neighbors by various chemotaxonomic and phenotypic properties. The results of the polyphasic analyses suggested that strain 13-Q(T) should be considered to represent a novel species of the genus Pedobacter, for which the name Pedobacter hainanensis sp. nov. is proposed. The type strain is 13-Q(T) (=CCTCC AB 2012076(T) = NRRL B-59850(T)).

Individualized radiation dose control in 256-slice CT coronary angiography (CTCA) in retrospective ECG-triggered helical scans: using a measure of body size to adjust tube current selection.

PURPOSE: To reduce radiation dose for retrospective ECG-triggered helical 256-slice CTCA by determining an optimal body size index to prospectively adjust tube current. METHODS: 102 consecutive patients with suspected CAD underwent retrospective ECG-triggered CTCA using 256-slice CT scanner. Six body size indexes including BMI, nipple level (NL) bust, thoracic anteroposterior diameter at NL, chest circumference (CC) at NL, left main and right coronary artery (RCA) origin level were measured and their correlation with noise was evaluated using linear regression. An equation was developed to use this index to adjust tube current. Additional 102 consecutive patients were scanned with the index-based mAs adjustment. A t-test for independent samples was used to compare radiation dose levels with and without the index-based mAs selection method. RESULTS: Linear regression indicated that CC RCA had the best correlation with noise (R2=0.603). Effective radiation dose was reduced from 16.6±0.9 to 9.8±2.7 mSv (p<0.01), i.e. 40.9% lower dose with the CC RCA-adapted tube current method. The image quality scores indicated no significant difference with and without the size-based mAs selection method. CONCLUSION: An accessible measure of body size, such as CC RCA, can be used to adapt tube current for individualized radiation dose control.

Evaluation of image quality and radiation dose at prospective ECG-triggered axial 256-slice multi-detector CT in infants with congenital heart disease.

BACKGROUND: There are a limited number of reports on the technical and clinical feasibility of prospective electrocardiogram (ECG)-gated multi-detector computed tomography (MDCT) in infants with congenital heart disease (CHD). OBJECTIVE: To evaluate image quality and radiation dose at weight-based low-dose prospectively gated 256-slice MDCT angiography in infants with CHD. MATERIALS AND METHODS: From November 2009 to February 2010, 64 consecutive infants with CHD referred for pre-operative or post-operative CT were included. All were scanned on a 256-slice MDCT system utilizing a low-dose protocol (80 kVp and 60-120 mAs depending on weight: 60 mAs for ≤ 3 kg, 80 mAs for 3.1-6 kg, 100 mAs for 6.1-10 kg, 120 mAs for 10.1-15 kg). RESULTS: No serious adverse events were recorded. A total of 174 cardiac deformities, confirmed by surgery or heart catheterization, were studied. The sensitivity of MDCT for cardiac deformities was 97.1%; specificity, 99.4%; accuracy, 95.9%. The mean heart rate during scan was 136.7 ± 14.9/min (range, 91-160) with a corresponding heart rate variability of 2.8 ± 2.2/min (range, 0-8). Mean scan length was 115.3 ± 11.7 mm (range, 93.6-143.3). Mean volume CT dose index, mean dose-length product and effective dose were 2.1 ± 0.4 mGy (range, 1.5-2.8), 24.7 ± 5.9 mGy·cm (range, 14.7-35.8) and 1.6 ± 0.3 mSv (range, 1.1-2.5), respectively. Diagnostic-quality images were achieved in all cases. Satisfactory diagnostic quality for visualization of all/proximal/distal coronary artery segments was achieved in 88.4/98.8/80.0% of the scans. CONCLUSION: Low-dose prospectively gated axial 256-slice CT angiography is a valuable tool in the routine clinical evaluation of infants with CHD, providing a comprehensive three-dimensional evaluation of the cardiac anatomy, including the coronary arteries.

Chitosan oligosaccharides suppress LPS-induced IL-8 expression in human umbilical vein endothelial cells through blockade of p38 and Akt protein kinases.

AIM: To investigate whether and how COS inhibited IL-8 production in LPS-induced human umbilical vein endothelial cells (HUVECs). METHODS: RT-PCR, enzyme-linked immunosorbent assays (ELISA) and Western blotting were used to study IL-8 expression and related signaling pathway. Wound healing migration assays and monocytic cell adhesion analysis were used to explore the chemotactic and adhesive activities of HUVECs. RESULTS: COS 50-200 µg/mL exerted a significant inhibitory effect on LPS 100 ng/mL-induced IL-8 expression in HUVECs at both the transcriptional and translational levels. In addition, COS 50-200 µg/mL inhibited LPS-induced HUVEC migration and U937 monocyte adhesion to HUVECs in a concentration-dependent manner. Signal transduction studies suggest that COS blocked LPS-induced activation of nuclear factor-κB (NF-κB) and activator protein-1 (AP-1) as well as phosphorylation of p38 mitogen-activated protein kinase (MAPK) and phosphokinase Akt. Further, the over-expression of LPS-induced IL-8 mRNA in HUVECs was suppressed by a p38 MAPK inhibitor (SB203580, 25 µmol/L) or a phosphatidylinositol 3-kinase (PI3K) inhibitor (LY294002, 50 µmol/L). CONCLUSION: COS inhibited LPS-induced IL-8 expression in HUVECs through the blockade of the p38 MAPK and PI3K/Akt signaling pathways.

Tantalum compounds as heterogeneous catalysts for saccharide dehydration to 5-hydroxymethylfurfural.

A new solid acid, based on tantalum hydroxide, was used to catalyze saccharide dehydration into 5-hydroxymethylfurfural (HMF) with high catalytic activity and excellent stability in a water-2-butanol biphasic system. Furthermore, good results were also obtained from Jerusalem artichoke juice with the catalyst under the same conditions.

Conversion of biomass into 5-hydroxymethylfurfural using solid acid catalyst.

5-Hydroxymethylfurfural (HMF) was produced from monosaccharide (fructose and glucose), polysaccharide (inulin) and the Jerusalem artichoke juice by a simple one-pot reaction including hydrolysis and dehydration using solid acid under mild condition. Hydrated niobium pentoxide (Nb(2)O(5)·nH(2)O(2)) after pretreatment showed high catalytic activities for dehydration of mono- and polysaccharide to HMF at 433 K in water-2-butanol (2:3 v/v) biphasic system, giving high HMF yield of 89% and 54% from fructose and inulin, respectively. The HMF yield was up to 74% and 65% when inulin and Jerusalem artichoke juice were hydrolyzed by exoinulinase. The solid acid made the process environment-friendly and energy-efficient to convert carbohydrates into bio-fuels and platform chemicals.

Chitosan oligosaccharides inhibit the expression of interleukin-6 in lipopolysaccharide-induced human umbilical vein endothelial cells through p38 and ERK1/2 protein kinases.

Chitosan oligosaccharides (COS) have been reported to exert anti-fungal activities, antitumour activities and immuno-enhancing effects. However, the potential roles of COS in the treatment of vascular inflammations remain unknown. In the present study, we examined the effects of COS on interleukin-6 (IL-6) production in human umbilical vein endothelial cells (HUVECs) induced by lipopolysaccharide (LPS). Induction of HUVECs with LPS (100 ng/ml) increased the mRNA expression and protein secretion of IL-6 (versus the vehicle-treated group, p < 0.01), which were significantly reverted by the pre-treatment with COS (50-200 microg/ml) for 24 hr before LPS exposure (versus the LPS-treated group, p < 0.05 or 0.01). Signal transduction studies showed that the pre-treatment of HUVECs with COS (50-200 microg/ml) for 24 hr markedly inhibited the LPS-induced over-expression of phosphorylated p38 mitogen-activated protein kinase (MAPK), phosphorylated ERK1/2 and nuclear factor kappaB (NF-kappaB). Moreover, the LPS-induced NF-kappaB activation was suppressed by the specific ERK1/2 inhibitor PD98059 (30 microM) (versus the LPS-treated group, p < 0.01), but not by the specific p38 MAPK inhibitor SB203580 (25 microM). Additionally, both MAPK inhibitors markedly suppressed LPS-induced IL-6 mRNA expression in HUVECs (versus the LPS-treated group, p < 0.01). In conclusion, our results suggest that COS inhibit LPS-induced up-regulation of IL-6 in HUVECs, and this can be regulated by at least two parallel signalling pathways: one via p38 MAPK pathway independent of NF-kappaB activation and one via ERK1/2 pathway dependent on NF-kappaB activation.